Dissertations/Thesis

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2024
Dissertations
1
  • WÉSLEI ALMEIDA COSTA ARAÚJO
  • Diagnostic evaluation of Strongyloides stercoralis infection in asthmatic individuals using recombinant antigens

  • Advisor : NECI MATOS SOARES
  • COMMITTEE MEMBERS :
  • ALVARO AUGUSTO SOUZA DA CRUZ FILHO
  • CARINA CARVALHO DOS SANTOS
  • NECI MATOS SOARES
  • Data: Feb 20, 2024


  • Show Abstract
  • Strongyloides stercoralis is the main etiological agent of human strongyloidiasis, a neglected disease with wide worldwide distribution. The infection is generally chronic and asymptomatic. However, in immunocompromised individuals using systemic glucocorticoids, such as individuals with severe asthma, S. stercoralis infection can progress to severe forms, hyperinfection and/or dissemination, with a low therapeutic response and a high lethality rate. Currently, the definitive diagnosis of S. stercoralis infection is made by searching for larvae in feces. However, this parasite releases few larvae and intermittently, which makes it necessary to examine several fecal samples. The objective of this study is to analyze the frequency of S. stercoralis infection in asthmatic individuals treated in the Asthma and Allergic Rhinitis Control Program (ProAR) in Bahia/Brazil, through parasitological and immunological diagnosis, using crude and recombinant antigens. A total of 500 asthmatics treated at ProAR were included in the study. The diagnosis of strongyloidiasis was made using three parasitological methods: spontaneous sedimentation (SE), Baermann-Moraes (BM) and, Agar Plate Culture (CPA). Serological diagnosis was preformed by IgG4 and IgE –ELISA using crude membrane antigen. Samples from individuals diagnosed positively by parasitological and /or immunological methods (n=66) were reevaluated through ELISA(s) using recombinant antigens (NIE and SsIR). Of the individuals studied, 21.2 (106/500) and 78.8% (394/500) were male and female, respectively. Around 42 (210/500) and 58% (290/500) of the individuals had mild/moderate or severe asthma, respectively, with 43.6% (218/500) being diagnosed in childhood. A frequency of enteroparasites was 8.6% (43/500) The most prevalent pathogenic parasite was S. stercoralis, 2.2% (11/500). The sensitivities of ELISA(s) for detection of IgG4 and IgE anti-S. stercoralis, using membrane antigen were 91.7% and 91.2%, respectively, and the specificities were 95.7% and 95.4%, respectively. Sera from individuals positive for S. stercoralis, diagnosed in this study, by finding larvae in feces and/or detection of IgG and IgE anti-S. stercoralis (n=66) were reevaluated through IgG-ELISA, using the recombinant antigens, NIE and SsIR, demonstrating sensitivities and specificities of 91.2, 85.3% and 95.4 and 90.9%, respectively. Immunoblotting confirmed the positivity of 39.4% (26/66) of reevaluated samples, revealing immunoreactivity bands of 30 to 90 KDa. Regarding the use of corticosteroids, 36.2% (181/500) reported having taken one or more cycles of oral corticosteroids for 3 or more days in 1 year and 8.4% (42/500) one or more cycles of injectable corticosteroids in 6 months. Among the 66 individuals who were positive (detection of larvae in feces for anti-S. stercoralis antibodies), 63.6% (42/66) used corticosteroids. The most, 60.6% (40/66) via inhalation, 30 .3% (20/66) via orally and 9.1% (6/66) via injection. The association observed between corticosteroid therapy and strongyloidiasis was not statistically significant (p = 0.768, 0.798, 0.351 and 1.0, respectively). The present study demonstrates that the agreement between the methods used was poor, encouraging new studies to develop an effective and reproducible test for the diagnosis of S. stercoralis infection. Furthermore, the study did not demonstrate a correlation between S. stercoralis infection and the use of glucocorticoids.

Thesis
1
  • CAROLINA DO ROSÁRIO ESTEVES GUIMARÃES
  • EVALUATION OF MOLECULES WITH EFFECT ANTIBIOFILM FOR Candida albicans
  • Advisor : TANIA FRAGA BARROS
  • COMMITTEE MEMBERS :
  • ANA PAULA DE OLIVEIRA MENEZES
  • ANIBAL DE FREITAS SANTOS JUNIOR
  • MARCELO SANTOS CASTILHO
  • TANIA FRAGA BARROS
  • YGOR JESSE RAMOS DOS SANTOS
  • Data: Feb 5, 2024


  • Show Abstract
  • Biofilms are highly organized communities of microorganisms that are less susceptible to antimicrobials than planktonic cells. Their development comprises one of the main virulence factors of Candida albicans, one of the most isolated and studied species in infections caused by Candida sp. When associated with biofilms, the treatment of these infections becomes more complicated due to the need for higher concentrations of antifungals than those required in infections caused by planktonic cells. Thus, therapeutic choices, which are limited, require new alternatives, of which biofilm inhibition and eradication appear to be promising strategies. Molecules with antibiofilm activity have been studied, but without taking care to use important criteria such as inhibition concentration of biofilm formation lower than the antifungal concentration and concentrations equal to or lower than 300 µM. In this sense, the aim of our study was to evaluate molecules with an antibiofilm effect on C. albicans, through a review of molecules described in the literature from 2011-2021 and the experimental analysis of two sets of molecules, one of thiosemicarbazones and analogues and the other of derivatives of natural products. In the literature review we discussed 21 articles, which described 42 molecules. Most of the molecules showed promise for inhibiting biofilm formation and had mechanisms of action that included changes in the adhesion process, yeast-hyphae transition, hyphae elongation, cell surface hydrophobicity and the production of quorum sensing components. These results guided the experimental part of the work, as well as the choice of some of the molecules tested. For the set of thiosemicarbazones and analogues innovated in the review, molecule 28 stood out for its greater potency in inhibiting biofilm formation by 50% (BIC50 = 31.55 ± 1.18 µM). For derivatives of natural products with chemical groups similar to those described in the review, molecule 6, identified as jatrophone, stood out for its greater inhibition power (BIC50 = 64.36 ± 1.05 µM) and ability to eradicate biofilms (BEC50 = 215.90 ± 1.07 µM). Scanning electron microscopy (SEM) studies showed that both molecules reduced the number of adhered cells, hyphae and pseudohyphae formed after exposure to BIC50, while jatrophone reduced the formation of hyphae in preformed biofilm in the presence of BEC50. Thus, we were able to determine the family of thiosemicarbazones and analogues, as well as jatrophone, as models for future studies that will allow us to further elucidate the mechanism of action of these molecules on the biofilm and optimise their structure to increase potency and selectivity.

2
  • LUÍZA CAROLINA FRANÇA OPRETZKA
  • Investigation of the therapeutic potential and patent analysis of two mesenchymal stem cell lines and their extracellular vesicles for neuropathic pain

  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CARLOS AMILCAR PARADA
  • CRISTIANE FLORA VILLARREAL
  • LUIZ FERNANDO FERRARI
  • SIMONE GARCIA MACAMBIRA
  • VICTOR DIOGENES AMARAL DA SILVA
  • Data: Feb 29, 2024


  • Show Abstract
  • INTRODUCTION: Neuropathic pain is a chronic pain syndrome that affects a significant proportion of the population and has devastating effects on patients' quality of life and work activity. Although there are therapeutic options, most patients benefit little or do not respond to treatment. Therefore, regenerative therapies with mesenchymal stem cells and their derivatives may help to fill this therapeutic gap.  OBJECTIVES: This study aimed to evaluate the therapeutic potential of a human mesenchymal stem cell line (MSC), an MSC-derived line overexpressing the leukemia inhibitory factor (MSC-LIF) and their extracellular vesicles ( VE-MSC and VE-LIF) in a model of neuropathic pain. It also aimed to explore the mechanisms underlying MSC’s and VE-MSC’s antinociceptive effects. Additionally, we aimed to perform a patent mapping focusing on MSC’s derivatives. METHODS: MSCs obtained from a biobank were genetically modified to overexpress LIF and characterized by flow cytometry and in vitro cell differentiation assays. The extracellular vesicles were isolated by ultracentrifugation and characterized by transmission electron microscopy, nanoparticle tracking analysis, and Dotblot. Cytokines released by macrophages stimulated in vitro and treated with MSC, EV-MSC, MSC-LIF, or EV-LIF were quantified by ELISA. Using the model of neuropathic pain induced by partial ligation of the sciatic nerve in C57Bl/6 mice, the antinociceptive effect of all treatments was evaluated by Hargreaves and von Frey filaments tests. Gait function was assessed on a treadmill. Knockout mice for interleukin-10 (IL-10 KO) and pharmacological antagonism assays with a selective CXCR2 antagonist (SB225002; 1 mg/kg, i.p.) were used to investigate the mechanisms of action of MSC and EV-MSC. Sections of spinal cord (L4-L5) and serum from mice were collected for cytokine and chemokine analysis by ELISA. Additionally, DWPI data base was used to retrieved the patentes. RESULTS:  Macrophages treated with MSC, EV-MSC, MSC-LIF, or EV-LIF showed reduced levels of TNFα, while IL-10 production was increased, indicating the possible application in sensory neuropathy. A single injection of MSC (1x106), EV-MSC (7.45x109 ± 2.25 x108 particles/mL), MSC-LIF (1x106) and EV-LIF (2.47 x109 ± 7.13 x107 particles/mL) provided complete and long-lasting relief of thermal hypernociception and improvement of gait functional parameters. Treatment with MSC and VE-MSC also completely relieved the mechanical hypernociception associated with neuropathy. Conversely, treatment with MSC-LIF and EV-LIF transiently and partially reduced the mechanical nociceptive behaviors of neuropathic mice. The participation of IL-10 in the therapeutical effect of MSC and EV-MSC was demonstrated since their antinociceptive effect was only partial in IL-10 KO mice compared to wild animals. In addition, IL-10 levels were elevated in the spinal cords of mice 14 days after treatment with MSC and EV-MSC but not at the end of the experimental period. In addition, acute treatment with SB225002 partially reversed the thermal antinociceptive effect of MSC and EV-MSC, while this antagonist completely reversed mechanical antinociception. However, serum levels of CXCL1, a CXCR2 ligand, did not differ between the experimental groups 14 days after treatment with MSC or EV-MSC and were elevated only in neuropathic animals at the end of treatment.  Regarding the patent search, 150 families of patents protecting mesenchymal stem cell’s derivatives were found, most of which were dedicated to the protection of extracellular vesicles and exosomes. There was an exponential increase in these patents from 2015 onwards. China was the country with the highest number of approved patents, followed by the United States. CONCLUSION:  The effect profile of stem cells and extracellular vesicles was quite similar, showing that extracellular vesicles can mediate the effect of mesenchymal stem cells, even those from genetically modified cells. In addition, the improvement of both nociceptive and functional parameters by treatment with MSC and EV-MSC reinforces the proposal for future application of these therapies for the treatment of neuropathic pain. Although the genetic modification of MSC via overexpression of LIF has been deleterious to the antinociceptive effect of MSC, this work has advanced the understanding of MSC’s mechanism of action in neuropathic pain, which seems to involve increased production of the anti-inflammatory cytokine IL-10 and to be maintained by the activation of CXCR2 receptors. Finally, patent analysis also indicated a growing interest of research and development of cell-free therapy products. Thus, this work corroborates the potential of cell-free therapy with MSC extracellular vesicles and its application in neuropathic pain.

2023
Dissertations
1
  • Izamir Resende Junior Borges Miguel
  • USE OF SUSTAINABLE RAW MATERIALS TO OBTAIN BIOMATERIALS: DENDROPHYLID CORALS (ANTHOZOA: SCLERACTINIA: DENDROPHYLLIDAE) AS A SOURCE OF CALCIUM PHOSPHATE
  • Advisor : RENATA BIEGELMEYER DA SILVA RAMBO
  • COMMITTEE MEMBERS :
  • ADEMIR EVANGELISTA DO VALE
  • ELIANA CRISTINA DA SILVA RIGO
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Jan 25, 2023


  • Show Abstract
  • In this study, a method was developed for the chemical conversion of aragonite skeletons from scleractinian corals (known as 'sol-coral'), non-native organisms introduced to the Brazilian coast and recorded for the Baía de Todos-os-Santos (BTS), to the development of applicable biomaterials such as bone grafts, having as an initial step the conversion of calcium carbonate into calcium phosphate. Scleractinian corals are bioconstructive organisms, which throughout life secrete a calcareous matrix in the form of a porous exoskeleton – indeed, inorganic (aragonite) and organic (tissues) parts define the architecture of these organisms. From coral carbonate, porous microstructures of complex architecture, and of biomedical interest, were obtained through chemical reactions under controlled temperature and constant agitation. Both, the raw material and the resulting biomaterials had their morpho-physicalchemical characteristics evaluated by a series of qualitative analyses, including: (1) X-ray powder diffraction (XRD) analysis, (2) transformed infrared spectroscopy (FTIR), (3) scanning electron microscopy (SEM), and (4) electron dispersive analysis (EDS). Based on the evaluation criteria adopted, and on the characteristics of imported bioceramics currently available, it is observed that the biomaterial produced in this research, in addition to being biomimetic and biphasic, has conditions to attend the prerequisites of compatible biocompatibility, osteoconduction and bioresorption expected in cases of bone regeneration. Additionally, due to the product's peculiar microstructure, potential application as a drug carrier is observed. Thus, the present work offers the development of national scaffolds, in the process of patenting, with great expectation of meeting the demands of the Unified Health System (SUS).

2
  • Daniela da Silva Nascimento
  • ASSOCIATION OF CEFTAZIDIME/AVIBACTAM AND AZTREONAM AGAINST METAL-β-LACTAMASE PRODUCING BACTERIA: EVALUATION OF THREE IN VITRO SYNERGY METHODS

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • CYNARA GOMES BARBOSA
  • JOICE NEVES REIS PEDREIRA
  • MARCELO PILONETTO
  • Data: Mar 2, 2023


  • Show Abstract
  • Resistance in Gram-negative bacteria is a global health concern due to the lack of viable therapeutic alternatives. Ceftazidime-avibactam (CAZ/AVI) is a combination of a third-generation cephalosporin and a β-lactamase inhibitor that has in vitro activity against serine-type enzymes such as extended spectrum β-lactamase (ESBL) and Klebsiella pneumoniae carbapenemase (KPC) class A, as well as class C cephalosporinases and class D OXA-48, but is ineffective against class B metallo-β-lactamases (MβL) producers. Aztreonam (ATM) is a monobactam that stands against MβL but is inactivated by ESBL, KPC and other cephalosporinases often found in stable preparations produced from MβL. The combined therapy of CAZ/AVI and ATM has been shown to be useful against these microorganisms. However, standardized commercial susceptibility tests for this combination are not available. Thus, testing in the laboratory the synergistic potential of the combination in vitro can guide the choice of an appropriate therapy. This study aims to compare the performance of three methods as predictors of synergistic activity between CAZ/AVI and ATM, in order to find a method that is low cost and easy to perform in a clinical microbiology laboratory. For this, a total of 36 MβL-producing bacteria were evaluated and the synergy test was performed with 30 isolates that showed CAZ/AVI and ATM resistance. Seven isolates were selected for the time-kill assay (TKA). β-lactam resistance genes were detected using the conventional PCR technique. The search for NDM variants was performed using Sanger sequencing. To determine the synergy between the drugs, disk-approximation (DA), overlapping diffusion gradient strips and TKA methods were used, the gold standard for synergy tests. The minimum inhibitory concentration (MIC) was performed by broth microdilution (MIC). Agreement (Kappa index) was calculated as a ratio of concordant responses between the evaluated methods and the TKA. The thirty-six samples tested carried blaNDM, and 8 (22%) had two metallo β-lactamases (blaNDM and blaVIM). Thirty-four isolates (94.4%) were detected as coproducers of metallo and serine β-lactamase, with 10 (27.7%) coproducers of blaKPC. The NDM-1 variant was detected in 31 (86.1%) samples. Thirty-five samples (97.2%) were considered multidrug resistant. The DA and diffusion gradient tape overlay methodologies agree very well (100%, κ = 1.0, p<0.05). Compared to TKA, the tested methodologies demonstrated sensitivity, specificity, positive predictive value, and negative predictive value of 100%. Twenty-nine (96.7%) showed a synergistic result of the ATM plus CAZ/AVI association so that the addition of avibactam was able to restore sensitivity to aztreonam, meeting MICs greater than 256 ug/mL for MICs less than 1 ug/mL. Thus, with this work it was possible to standardize two possible methodologies to be used in the daily routine of the clinical microbiology laboratory to predict in vitro the synergy of the association of ATM plus CAZ/AVI in isolates of K. pneumoniae producing metallo-β-lactamase

3
  • GLEISEQUELLE OLIVEIRA DOS SANTOS SOUZA
  • Pre-malignant and malignant lesions of the uterine cervix: diagnosis, follow-up and treatment following the guidelines recommended by the Brazilian Ministry of Health

  • Advisor : JUNIA RAQUEL DUTRA FERREIRA
  • COMMITTEE MEMBERS :
  • CLÁUDIA MARTINS CARNEIRO
  • PEDRO COSTA CAMPOS FILHO
  • JUNIA RAQUEL DUTRA FERREIRA
  • Data: Apr 10, 2023


  • Show Abstract
  • Objective: To evaluate the referral, diagnosis, follow-up and treatment of patients with abnormal cervical cytology to a Medium-Complexity Unit (MCU) following the guidelines recommended by the Brazilian Ministry of Health. Methods: This is a retrospective descriptive study, based on abnormal cervical cytology of users of the Unified Health System, city of Salvador, Bahia, from January 2017 to January 2020. Methods: This is a retrospective descriptive study, developed at the Women's Hospital, Salvador, Bahia, where data were collected from electronic medical records, from January 2017 to January 2020. The variables as: abnormal cervical cytology results (pre-malignant and malignant squamous lesions), colposcopic, histopathological and clinical follow-up as well as applied treatment methods were collected. Data analysis was performed with IBM SPSS 20. Results: 908 patients were referred to the MCU. Of the total, 812 (89,4%) were in the age according to the guidelines of cervical cancer screening. Most patients were classified as more severe squamous lesions (ASC-H/HSIL/CEC), while 21.7% were classified as less severe squamous lesions (ASC-US/LSIL). CIN II/III was the most observed histopathological diagnosis in the study. Follow-up of repeat cytological was more observed in the group with less severe squamous lesions, while LEEP was the most used procedure in the treatment of more severe squamous lesions. Of patients underwent excisional procedure, most (71.6%) treated with LEEP and 132 (28.4%) treated with CKC. Compared to LEEP, patients undergoing CKC were significantly younger (p=0.019). Associations were observed regarding margin evaluation (p<0.001), glandular involvement (p<0.001). As for the presence of residual lesion, parameters such as age ≥35 years old (p=0.028) and margin status (p<0.001) were statistically significant in association with lesion persistence. Conclusion: Although most referrals was in accordance with Brazilian Ministry of Health recommendations, there is a significant frequency of inappropriate and unnecessary referrals to a MCU for colposcopy and biopsy. Regarding residual lesion, age ≥35 years and positive margin may be associated with persistence of CIN II/III.

4
  • ANA MARIA LENZ CARDOSO
  • DEVELOPMENT OF A SINGLE-STEP DRYING PROCESS FOR POLYMERIC NANOPARTICLES TO OBTAIN DERMATOLOGICAL PRODUCTS

  • Advisor : HENRIQUE RODRIGUES MARCELINO
  • COMMITTEE MEMBERS :
  • HENRIQUE RODRIGUES MARCELINO
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • RUY CARLOS RUVER BECK
  • Data: Jul 20, 2023


  • Show Abstract
  • The work presented here is divided into two parts. Chapter 01 is a bibliographic review in which the literature of the last years (2016 - 2022) was examined to evaluate the development of nanoparticle formulations with Eudragit® types L 100, S 100, RL 100 and RS 100 for topical application. The second chapter contains the experimental part, where the development of a process to obtain Eudragit® L 100 nanospheres in a single step and the application of these formulations in a semi-solid carrier for topical application was proposed, studying the physicochemical properties, rheology and texture.

5
  • JACKELINE MARLEY SANTOS DE ARAÚJO
  • Physiology based pharmacokinetic modeling of methylphenidate in adults

  • Advisor : FRANCINE JOHANSSON AZEREDO
  • COMMITTEE MEMBERS :
  • ANDREA DINIZ
  • FRANCINE JOHANSSON AZEREDO
  • IZABEL ALMEIDA ALVES
  • Data: Nov 21, 2023


  • Show Abstract
  • Introduction: Methylphenidate (MPH) is a psychotropic stimulant drug used as first pharmacological choice for treating Attention/Hyperactivity Deficit Disorder (ADHD). However, problems related to their use make further investigation of their pharmacokinetics (PK) in treating ADHD in the adult population needed. Physiology-based Pharmacokinetic modeling (PBPK) is a mathematical approach that aims to simulate concentration profiles versus time for different administration pathways integrating the physiological structure of a particular species and physicochemical characteristics of a compound. PK-Sim® is free open-source software developed by Open Systems Pharmacology (OSP) to construct full-body PBPK models. It can be used to simulate different species, characterizing their respective organisms in different biological compartments. Objective: The central objective of this work was to evaluate the variability of exposure after the use of MPH in adults. Methodology: A PBPK model for MPH was developed in PK-Sim® software (version 11.2) using information obtained from Literature on MPH, adult individuals, and two types of formulations. Results and Discussion: A PBPK model has been developed and validated whose predictive performance adequately meets the predefined criteria PK characteristics in Caucasian individuals and in a Caucasian population using a modified release formulation. Complementarily, a PK-Sim®-based tutorial was developed to enable readers to perform a PBPK analysis to provide a mechanistic approach to study and predict the PK of compounds based on the physiological and anatomical characteristics of a population of interest, as well as in the physical and chemical properties of a certain drug, as well as encouraging the production and teaching of Pharmaceutical Sciences among Portuguese speakers. Conclusion: This study properly verified the variability of exposure to MPH use in human adults. In addition, it also produced a Portuguese tutorial guiding PBPK modeling using PK-Sim®.

6
  • AYLA WINNIE RAMOS DA SILVA
  • Chemical analysis of Erythrina velutina alkaloids and toxicological evaluation in zebrafish model.

  • Advisor : RENATA BIEGELMEYER DA SILVA RAMBO
  • COMMITTEE MEMBERS :
  • ADEMIR EVANGELISTA DO VALE
  • NEILA DE PAULA PEREIRA
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Dec 5, 2023


  • Show Abstract
  • The use of natural products to treat diseases is an ancient custom in various cultures and peoples, and as knowledge develops, more is discovered by science about the mechanism of action of these compounds. Plants of the genus Erythrina have been highlighted in research for their varied pharmacological activities, mainly related to the Central Nervous System, such as their anxiolytic and anticonvulsant action. Although several studies address the phytochemical analysis of these species, the standardization of this analysis for each species is very necessary, with a view to better directing the studies. The isolation of erythrin alkaloids can be used to verify the specificity and potency of their pharmacological properties; The methods for carrying out this procedure are varied, with emphasis on the application of chromatographic methods. However, the development of new drugs is hampered by the lack of models that respond to biological activities in a reliable and less expensive way. In this sense, zebrafish stand out, due to their size, transparency, easy handling and rapid development, in addition to their physiological similarity to mammals, including humans. For a better performance of these studies, establishing a safe dose is an important step, carrying out what has been called a toxicity test, where newly fertilized embryos are exposed to the compound or set of compounds under analysis for 96h, in serial concentrations. In this sense, this work aimed at chemical analysis by HPLC of the fraction enriched in alkaloids from E. velutina, with a view to determining its best collection time, as well as the toxic potential of this fraction in an in vivo study. This analysis found the month of October to be the most suitable for investigating the composition of the extracts, due to the number of qualifying chromatographic peaks presented, considering their areas. The toxicity test with leaf, flower, stem bark and root bark extracts yielded LC50 values of 187.9 ± 1.47 µg/mL; 358.4 ± 1.25 µg/mL; 338.3 ± 1.25 µg/mL, and; 233.1 ± 1.86 µg/mL, respectively. Additionally, considering the potential of erythrin alkaloids for the treatment of central nervous system disorders, a systematic review of the application of the zebrafish model for screening natural products on this topic was carried out. 

7
  • MATHEUS ANTÔNIO DA HORA BORGES
  • BIOANALYTIC VALIDATION, PHARMACOKINETIC ANALYSIS AND PREPARATION OF AN α BISABOLOL NANOEMULSION

  • Advisor : FRANCINE JOHANSSON AZEREDO
  • COMMITTEE MEMBERS :
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • FRANCINE JOHANSSON AZEREDO
  • SANDRA ELISA HAAS
  • Data: Dec 7, 2023


  • Show Abstract
  • Natural products are the greatest basis for the discovery and development of new drugs. Chamomile is an herb commonly used in the preparation of teas by different cultures and has several reported therapeutic effects. α-bisabolol (α-BIS), its main constituent, has been the target of several pharmacological studies due to its ability to act on different body systems, from inflammatory processes and pain modulation to the fight against microorganisms. Despite knowledge about their effects, the behavior of drugs in the body has not yet been elucidated, as there are no pharmacokinetic studies on them. Knowing pharmacokinetics makes it easier to understand the pharmacodynamic mechanisms in which they participate. For pharmacokinetic analysis, a bioanalytical method is necessary so that it is possible to quantify plasma concentrations of the compound after administration. Therefore, the objective of this work was to develop and validate a bioanalytical method for the determination of α-bisabolol in plasma using High-Performance Liquid Chromatography (HPLC) and subsequently evaluate its pharmacokinetics. To achieve this, the method was developed in accordance with the requirements recommended by the Food and Drug Administration (FDA) and the Agência Nacional de Vigilância Sanitária (ANVISA). Chromatographic conditions consisted of an isocratic flow of 1 ml/min of acetonitrile and ultrapure water (80:20). The developed method proved to be precise, accurate, sensitive, and selective for quantifying the compound up to one month after freezing. To demonstrate the method’s applicability, a pharmacokinetic analysis of α-BIS was performed after intravenous administration of the compound in Wistar rats. After this administration, pharmacokinetic parameters such as volume of distribution, clearance, and half-life. With this validated method and pharmacokinetic analysis carried out, it becomes possible to develop new in vivo studies that aim to quantify α-bisabolol in biological matrices, in addition to assisting in new pharmacodynamic studies, since, for the first time to date, the pharmacokinetics of the compound were described. 

Thesis
1
  • Pedro Santana Sales Lauria
  • Preclinical characterization of ayahuasca's antinociceptive properties

  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CARLOS AMILCAR PARADA
  • TIAGO ARRUDA SANCHEZ
  • CRISTIANE FLORA VILLARREAL
  • DANIEL PEREIRA BEZERRA
  • RENAN FERNANDES DO ESPIRITO SANTO
  • Data: Jul 12, 2023


  • Show Abstract
  • Neuropathic pain affects a significant portion of the global population. The clinical management of neuropathic pain is challenging due to the low efficacy of currently available treatments, which motivates the search for new therapeutic options. Several studies show that psychedelics promote analgesic effects in different chronic pain conditions. Ayahuasca (AYA) is a psychedelic brew used by many religious groups worldwide. The therapeutic potential of AYA is well-documented in the treatment of psychiatric conditions such as depression and drug addiction. However, despite the anecdotal evidence that AYA promotes analgesic effects in the religious context, this effect is still poorly studied. Therefore, this study aimed to investigate and characterize the antinociceptive effects of AYA. The antinociceptive effect of oral treatments with AYA was assessed in male Swiss or C57BL/6 mice in the formalin test, Complete Freund's Adjuvant (CFA) model of inflammation, tail flick test, and partial sciatic nerve ligation model of neuropathic pain. Motor coordination and spontaneous locomotion were assessed in the rota-rod and open field tests, respectively. Possible mechanisms of antinociception were investigated by assays of pharmacological antagonism and immunohistochemically assessing Fos expression in brain areas that modulate nociception. Parameters suggestive of systemic toxicity were investigated following acute or multiple exposures to AYA. Chemical characterization of AYA was made by HPLC and the antinociceptive effect of its major component, harmine, was tested in experimental neuropathy. AYA (24 - 3000 μL/kg) dose-dependently reduced formalin-induced pain-like behaviors and CFA-induced mechanical allodynia but did not affect CFA-induced paw edema or tail flick latency. During experimental painful neuropathy, single treatments with AYA (24 - 3000 μL/kg) reduced mechanical allodynia; daily treatments once or twice a day for 14 days promoted consistent and sustained antinociception. The antinociceptive effect of AYA (600 μL/kg) was reverted by bicuculline (GABAA receptor antagonist; 1 mg/kg) and methysergide (non-selective serotonergic antagonist; 5 mg/kg), but not by naloxone (non-selective opioid antagonist; 5 mg/kg), phaclofen (GABAB receptor antagonist; 2 mg/kg), and rimonabant (CB1 inverse agonist; 10 mg/kg), suggesting the role of GABAA and serotonergic receptors in AYA-induced antinociception. AYA increased Fos expression in the ventrolateral periaqueductal gray and nucleus raphe magnus 1 h after the treatment, but not after 6 h or 14 days of daily treatments. AYA (600 μL/kg) acutely or twice a day for 14 days did not alter mice’s motor function, spontaneous locomotion, body weight, food and water intake, hematological, biochemical, and histopathological parameters. Harmine (3.5 mg/kg, orally), the major component present in AYA, promoted consistent antinociception during experimental neuropathy. Taken together, the results of this study allow the conclusion that AYA promoted consistent antinociceptive effects in different mouse models of pain without inducing detectable toxic effects. Harmine is at least partially accountable for the antinociceptive properties of AYA.

2
  • Suellen Laila Rocha Silva
  • Study of the anti-leukemic potential of emetine in eliminating leukemic stem cells from acute myeloid leukemia
  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • RAQUEL CARVALHO MONTENEGRO
  • PAULO MICHEL PINHEIRO FERREIRA
  • CYNARA GOMES BARBOSA
  • DANIEL PEREIRA BEZERRA
  • GARDENIA CARMEN GADELHA MILITÃO
  • Data: Jul 19, 2023


  • Show Abstract
  • INTRODUCTION: Acute myeloid leukemia (AML) is a disease with a low 5-year survival rate. One of the reasons for this high mortality is the resistance of leukemic stem cells (CTLs) to conventional treatments, leading to disease recurrence. The NFκB signaling pathway is activated in AML CTLs, making it an attractive target for the elimination of these cells. Emetine (EMT), an antiparasitic used clinically, has been shown to inhibit NF-κB signaling. OBJECTIVE: To evaluate the antileukemic potential of emetine in eliminating AML LSCs in an in vitro and in vivo translational model using KG-1a cells. METHODOLOGY: Initially, EMT was tested on a panel of cancerous and non-cancerous cells to determine the IC50 and evaluate its cytotoxicity. Then, the presence and elimination of CTLs was identified by flow cytometry, using antibodies for CD34, CD38, CD13, CD33 and CD123. Assays to investigate the mechanism of action of the compound using the KG-1a strain were performed, including trypan blue exclusion assay, analysis of cell death pattern, expression of caspase-3 and PARP-1, activation of the NF-κB pathway, evaluation of the cell cycle, oxidative stress and mitochondrial depolarization. In addition, analyzes were performed by confocal microscopy, qPCR and in vivo assay using NSG mice. RESULTS: EMT showed cytotoxicity against all cancer lines tested, with an IC50 of 0.74 µM for the KG-1a. The compound also reduced the percentage of CD13, CD34, CD38 and CD123 positive cells. It was observed that EMT induced apoptotic cell death, increased the expression of active caspase-3 and PARP-1 cleavage, promoted mitochondrial depolarization, generation of reactive oxygen species and internucleosomal DNA fragmentation, in addition to inhibiting the expression of NF- κB. Of the 92 genes analyzed by qPCR, 54 were up-regulated and 5 were down-regulated by the compound. In the xenograft model, EMT treatment reduced the amount of leukemic cells in the bone marrow and blood of mice. CONCLUSION: EMT has shown to be a promising cytotoxic compound, able to eliminate AML LSCs in vitro and in vivo, inducing the expression of active caspase-3, cleavage of PARP-1, generation of reactive oxygen species, fragmentation of internucleosomal DNA and inhibition of NF-κB pathway in KG-1a cells.

3
  • RAFAELA GOMES ALVES COSTA
  • PHARMACOLOGICAL STUDY OF BORTEZOMIB IN THE TREATMENT OF ACUTE MYELOID LEUKEMIA WITH ACTION ON STEM CELLS LEUKEMIC
  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • ANA JÉRSIA ARAÚJO
  • DALILA LUCIOLA ZANETTE
  • DANIEL PEREIRA BEZERRA
  • ELISANGELA VITORIA ADORNO
  • JEAN NUNES DOS SANTOS
  • Data: Jul 31, 2023


  • Show Abstract
  • INTRODUCTION: Acute myeloid leukemia (AML) is characterized by the uncontrolled growth of immature blasts in the bone marrow and corresponds to one of the most common types of cancer in the world. The 5-year relative survival rate is relatively low, and most patients eventually relapse. It is believed that one of the main factors responsible for the low rate of complete remission of the disease is associated with leukemic stem cells (CTL). Studies suggest that AML is originated and maintained by this population of CTLs. The NF-κB signaling pathway acts in the progression of cancer after tumor formation, being constitutively active in CTLs and, for this reason, it becomes an important therapeutic target. In this context, bortezomib (BTZ) is already described as a potent inhibitor of the NF-κB pathway and has been studied for the treatment of some hematological diseases. OBJECTIVE: Evaluate the antileukemic potential of BTZ as a therapeutic strategy to eliminate human AML CTLs in an in vitro and in vivo model. METHODOLOGY: BTZ cytotoxicity was tested on a panel of cancerous and non-cancerous cell lines. CTLs were identified using antibodies CD34, CD38, CD133, CD13 and CD123. The KG-1a strain was used to carry out BTZ mechanism of action assays in order to assess viability through cell cycle analysis, detection of cell death by apoptosis, evaluation of mitochondrial transmembrane potential and production of reactive oxygen species (ROS) by the flow cytometry technique. Analyzes of gene expression and signaling pathways associated with cancer through qPCR in cells treated with BTZ were also performed, in addition to assays for antitumor evaluation in an in vivo model. RESULTS: BTZ was cytotoxic to different strains tested with IC50 values ranging from 0.13 to 9.17 µM for cancer cell lines NB4 and U87 respectively. In addition, BTZ reduced the population of viable CTLs, induced apoptotic cell death, increased expression of active caspase-3 and cleaved PARP-1, induced alterations in mitochondrial transmembrane potential, increased ROS levels, induced DNA fragmentation in cells KG-1a and altered the gene expression of KG-1a cells, in addition to inhibiting the growth of these cells in an in vivo model. CONCLUSION: The antileukemic activity performed by BITZ emerges as a promising new path for the treatment of AML, with CTLs as the main target.

2022
Dissertations
1
  • ANA FLÁVIA SOUTO FIGUEIREDO NEPOMUCENO
  • New analytical strategy for determination of mefenamic acids in drugs by colorimetry of digital images. 

  • Advisor : LIZ OLIVEIRA DOS SANTOS
  • COMMITTEE MEMBERS :
  • LIZ OLIVEIRA DOS SANTOS
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • MARCOS DE ALMEIDA BEZERRA
  • Data: Feb 17, 2022


  • Show Abstract
  • The development of a new, simple, fast and low-cost analytical strategy for the determination of mefenamic acid in drug samples is described in this work. The proposal was based on the pre-concentration of mefenamic acid, using ultrasound-assisted liquid-phase microextraction (UA-LPME) and subsequent detection by digital image colorimetry (ID). The UA-LP with a eutectic solvent consisting of thymol and product (obtaining solvent) and a copper sulfate solution. Univariate optimization, optimal adjustment conditions such as working solvent samples, time adjustment and pH adjustment, adjustment solution type, solvent time adjustment, adjustment time adjustment and pH adjustment. Under optimized conditions, the new method had a detection limit of 0.43 µg/mL, a limit of quantitation of 1.31 µg, and an accuracy (relative standard deviation) of 2.38%. The method precision was made by means of comparison with the reference method of Ab or Atomic Spectrometry with Graphite Furnace at 95% confidence. In the analyzed samples, for 500 mg drugs, mean concentrations in the range of 499 to 532 mg of mefenamic acid were found. The proposed procedure was successfully applied to the determination of mefenamic acid in a sample of drugs sold in the city of Salvador, Bahia, Brazil. The method complies with the principles of environmental chemistry as it uses a small risk of green sample materials and according to the developed method generates a small risk of green sample materials and uses a low risk product model and according to the potential of ambient reagent.

2
  • MARIANA SILVA CARDOSO
  • Validation of analytical methodology for determination of HVA and 5- Urinary HIAA by HPLC, and its application in the evaluation of children exposed to emissions from a ferro-manganese alloy plant in Bahia, Brazil
  • Advisor : JOSE ANTONIO MENEZES FILHO
  • COMMITTEE MEMBERS :
  • ELIANI SPINELLI
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • JOSE ANTONIO MENEZES FILHO
  • Data: Apr 28, 2022


  • Show Abstract
  • Introduction: Vila de Cotegipe, located in Simões Filho, in the metropolitan area of
    Salvador, Bahia, has housed for almost 40 years, a ferro-manganese alloy production
    plant. Excessive exposure to this metal has been reported as neurotoxic for causing
    damage to specific regions of the brain related to dopamine (DA) metabolism and, in
    some cases, altering brain levels of serotonin (5-HT). In children, high levels of Mn are
    associated with neurological disorders such as behavioral changes and intellectual
    deficit.
    Objectives: The objectives of this study were (i) to develop and validate an analytical
    method for determining urinary metabolites of dopamine and serotonin, HVA, and 5-
    HIAA, respectively; (ii) to evaluate the exposure of schoolchildren to Mn by measuring
    manganese in hair (MnH) and manganese in toenails (MnTn) and the deposition rate of
    manganese in the school dust; (iii) and to investigate whether there is an association
    between the levels of these metabolites and the biomarkers of exposure to manganese.
    Method: The study was conducted with a group of children aged 6 to 12 years (n=30)
    residing in the vicinity of a ferro-manganese alloy plant in Vila de Cotegipe, Simões
    Filho, Bahia. For comparison purposes, 30 children in the same age group from the
    municipality of Aratuípe, also in the Recôncavo Baiano, were included (reference
    group). The levels of MnH, MnTn, and Mn in the dust were determined by graphite
    furnace atomic absorption spectrometry. For the determination of urinary 5-HIAA and
    HVA levels, a method was developed using solid-phase extraction columns with a
    strong ion exchanger sorbent. An analytical method by HPLC-ECD was validated.
    Results: The chromatographic method developed and validated showed satisfactory
    selectivity, sensitivity, precision, and accuracy according to the RDC No. 27/2012 of
    ANVISA. The limit of detection (LOD) was 4 µmol/L and 8 µmol/L for 5-HIAA and
    HVA, respectively, in natural urine. The recoveries were 85.8% and 92.3% for 5-HIAA
    and HVA, respectively, in low concentration, and 90.7% and 94% for 5-HIAA and
    HVA, respectively, at high concentration levels. The levels of urinary metabolites in the
    children from the exposed and reference groups were within the physiological values.
    The medians (minimum and maximum) for 5-HIAA and HVA of the exposed group
    9
    were 36.4 µmol/L (18.4 - 58.0) and 32.9 µmol/L (<LOD - 91.9), respectively. There
    was no statistically significant difference between the values presented by the children
    in the reference group: 25.7 µmol/L (19.9 - 81.4) and 35.2 µmol/L (<LOD -67.6) for 5-
    HIAA and HVA, respectively.
    The medians (minimum and maximum) for MnH and MnTn of the exposed
    group were 4.95 μg/g (0.31 - 21.94) and 14.32 μg/g (2.54 - 81.0), respectively. These
    values are approximately 50 and 120 times higher than in the reference group, whose
    medians were 0.10 μg/g and 0.12 μg/g, respectively. Stratifying by gender, girls had
    higher MnH values than boys, while these had higher MnTn values. The medians of the
    deposition rate of manganese in the dust of the school in Vila de Cotegipe were 52.194
    µg Mn/m2
    /30 days for the external area and 9.734 µg Mn/m2
    /30 days for the internal
    area.
    A strong positive correlation was observed between MnH and MnTn levels (rho =
    0.748; p-value < 0.01) and a weak inverse correlation between the exposure biomarkers
    (MnH and MnTn) and age in months (rho = -0.337; p-value < 0.05 and rho = -0.374; pvalue < 0.01, respectively). A weak positive and significant correlation (rho = 0.373)
    between 5-HIAA and HVA was observed. However, no significant correlation was
    observed between the levels of these analytes and the biomarkers of exposure to Mn
    (MnH and MnTn).
    Conclusions: The SPE cartridge with a strong ion exchanger was efficient in the
    extraction of urinary metabolites, enabling a good recovery. The chromatographic
    method developed and validated followed the parameters required to be applied, and the
    electrochemical detection proved to be a sensitive technique and a viable alternative for
    use in urinary HVA and 5-HIAA dosages. Although children living in Vila de Cotegipe
    are environmentally exposed to high levels of Mn, the urinary metabolite levels were
    within normal physiological values. We can conclude that the urinary excretion of the
    studied metabolites possibly, does not reflect the interference of Mn in the metabolism
    of dopamine and serotonin in the CNS

3
  • URI RAMOS FIRMO
  • Human Papilloma Virus infection in individuals with cancer of the oral cavity and oropharynx treated at a tertiary hospital in Salvador - Bahia

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • JOICE NEVES REIS PEDREIRA
  • CYNARA GOMES BARBOSA
  • MARCUS ANTONIO DE MELLO BORBA
  • Data: May 16, 2022


  • Show Abstract
  • Introduction: Cancer is one of the main causes of morbidity and mortality in the world, with carcinoma of the oropharynx and oral cavity representing about 2% to 4% of all cases, accounting for the fifth most common type of cancer in men in Brazil. Classically associated with lifestyle habits such as smoking and alcohol consumption, at an older age, the appearance of these cancers in younger individuals without such habits has been associated with Human Papillomavirus (HPV) infection. This virus with a wide global distribution and high prevalence may play an important role in the carcinogenesis of cancers of the oral cavity and oropharynx in our population, but there is no data in the literature that estimates the real importance of formal information on patients. Objective: To investigate HPV infection in patients diagnosed with cancer of the oral cavity and oropharynx treated at a tertiary hospital specialized in cancer treatment in the city of Salvador, Bahia, in the period 2017-2021. Method: A retrospective, descriptive observational study, with inclusion criteria of people residing in the state of Bahia who was referred for diagnosis, staging, and/or treatment of oropharyngeal or oral cavity carcinoma at Hospital Aristides Maltez (HAM) - Tertiary Hospital specialized in the treatment of cancer, over 18 years old, located in Salvador-BA, from 2017 to 2021. A search for p16 protein by immunohistochemistry, an indirect marker of HPV infection, was performed in lesions suspected and diagnosed with oral cavity and oropharyngeal cancer. Results: Among the 39 individuals who participated in the study, 25.6% (10) were considered HPV positive due to p16 positivity. HPV negative patients had a mean age of 63.0 years, while HPV positive patients had a mean age of 55.4 (p=0.03, Mann Whitney). A significant association was found between cancer staging and age in HPV-negative patients, showing that elderly patients are associated with more advanced stage tumors (r= 0.397; p= 0.04, Spearman). Regarding the association between alcoholism and smoking, it was found that 90.5% (19 individuals) of those who drink and smoke at the same time have more advanced tumor staging (p=0.03; Fisher's exact test). Discussion: Our study showed that the study population mostly has HPV negative oral cavity and oropharynx tumors (74.35%), with HPV positive tumors accounting for 25.65%, being more common in younger individuals. Smoking and drinking habits, concomitantly, are associated with more advanced grades of tumors. Conclusion: Most cases of the oral cavity and oropharyngeal cancer in our population are not associated with HPV. Its prevalence is similar to that of developed countries about 20 years ago, enabling the development of public policies to prevent HPV infection.

4
  • MÁRVIA CLEYSSE CUNHA CORREIA
  • EVALUATION OF THE CHEMICAL VARIABILITY OF CORALSOL SPECIES COLLECTED IN THE BAY OF ALL-OS-SANTOS

  • Advisor : RENATA BIEGELMEYER DA SILVA RAMBO
  • COMMITTEE MEMBERS :
  • ADEMIR EVANGELISTA DO VALE
  • ELIZABETH GERARDO NEVES
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Jun 30, 2022


  • Show Abstract
  • Corals of the genus Tubastraea (Cnidaria, Anthozoa) are native to the Pacific and Indian Oceans, being currently distributed along the tropical waters of the Atlantic, mainly on the Brazilian coast, due to the bioinvasion process. The colonies of Tubastraea are known as ‘Sun Coral' and the chemical defense present is an important strategy, being sustained by the production of antifouling, antipredation and release of allelopathic metabolites, due to the production of hydrocarbons, terpenes, steroids and alkaloides as the main classes of substances. Moreover, studies have demonstrating important pharmacological activities for alkaloids present in marine organisms, such as specific toxicity to cancer cells, antiplasmodial and antimicrobial activities, besides properties related to neurotransmission modulation. In this context, the present work was intendend to determine the chemical profile to evaluate the chemical variability of sun coral species collected at the Todos-os-Santos Bay (BA). The corals were collected at the Todos-osSantos Bay (BA), in different months between 2020 and 2021. The extracts were prepared using methanol as a solvent in the ratio of 1:3 w/v (coral:solvent). The colonies, after lyophilized, were extracted using ultrasound bath and then static maceration. The HPLC/DAD method was developed using luna® C18 column (5μm, 250 x 4.6 mm) with a flow rate of 0.7 mL/min. The mobile phase used was water: trifluoroacetic acid (TFA) (99.94:0.06 v/v;) (phase A) and methanol as phase B in a gradient elution. Analysis of Tubastraea chemical profile showed higher peaks at 24min, 27min and 30min with UV spectrum at wavelengths of 250nm, 277nm and 348nm, characteristic of aplisinopsin alkaloides. It concludes that the method developed for extraction and chemical analysis was effective for the objective proposed by the work. This work made it possible to establish a chemical profile for the sun coral in Todos-os-Santos Bay (BA), where we could observe the chemical variability between the types and collection sites. This similarity in the special metabolism of coral-sun species, where it was possible to establish a major compound as a possible chemical marker of the species, because it is repeated independently of the location and type of sample.

5
  • Elenita Bastos Almeida Costa
  • Chemical-pharmaceutical analysis and stability studies of estriol.
  • Advisor : EDITH CRISTINA LAIGNIER CAZEDEY
  • COMMITTEE MEMBERS :
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • HENRIQUE RODRIGUES MARCELINO
  • CRISTIANI LOPES CAPISTRANO GONCALVES DE OLIVEIRA
  • Data: Jul 29, 2022


  • Show Abstract
  • The present experimental research aimed to analyze, develop and validate analytical methods for the quality control of estriol active pharmaceutical ingredient (API) and dosage forms, cream 2 mg/g and tablet 1 mg, as well as to develop and validate an analytical method indicative of stability for the API and the finished products. Therefore, qualitative analysis of estriol was performed using spectrophotometry in the UV and IR regions, high-performance liquid chromatography, and other general methods to characterize API, vaginal cream, and estriol tablets. The techniques involving spectrophotometry in the UV region, zero-order and by derivative, respectively, were used to develop and validate quantitative methods for tablets and cream. The research revealed that all validation parameters met the main guides and compendia requirements. In addition, a forced degradation study was carried out for API and for the finished products in acidic, basic, and neutral media, under oxidizing conditions, in the presence of metal ions, and under photolytic conditions. The results obtained disclosed that, except for the photolytic medium, the formation of degradation products was observed under all other conditions. Furthermore, the work developed and validated quantitative methods indicative of stability by high-performance liquid chromatography using a diode array detector, C18 analytical column, with a mobile phase consisting of water (pH=4.0, adjusted with phosphoric acid) and ethanol (70:30, v/v), with a column oven temperature of 35 °C. Detection of the principal peak was performed at a wavelength of 280 nm and a flow rate of 1.0 mL/min.

6
  • HELEN REGINA SILVA SODRÉ DE MATOS
  • Assessment of exposure factors to SARS-CoV-2 in the professionals of the extension laboratories of the Faculty of Pharmacy - Federal University of Bahia, and monitoring of seroconversion to COVID-19

  • Advisor : MARCIA CRISTINA AQUINO TEIXEIRA
  • COMMITTEE MEMBERS :
  • ANTONIO RICARDO KHOURI CUNHA
  • LUCIANA SANTOS CARDOSO
  • MARCIA CRISTINA AQUINO TEIXEIRA
  • Data: Aug 19, 2022


  • Show Abstract
  • The COVID-19 pandemic caused by SARS-CoV-2 brought impacts to the entire population, especially to health professionals who were most exposed to the virus. Thus, knowledge of the prevalence of anti-SARS-CoV-2 antibodies among healthcare professionals is important to understand the spread of COVID-19 in the workplace. The objective of this work was to determine and monitor the frequency of SARS-CoV-2 infection in employees of the extension laboratories of the Faculty of Pharmacy of UFBA and the associated risk factors, through viral RNA and specific serum antibodies, at the return of their in-person activities during the pandemic. Eighty-five laboratory collaborators were monitored for 6 months for the detection of anti-SARS-CoV-2 IgG antibodies and viral RNA, before being vaccinated for COVID-19. Antibody detection was performed by serological tests, such as enzyme immunoassay (ELISA) and chemiluminescence, while viral RNA analysis was performed by real-time reverse transcription polymerase chain reaction (RT-qPCR). In addition, a questionnaire was applied to collect sociodemographic and risk factors for transmission of COVID-19 information. The seroprevalence for IgG anti-SARS-CoV-2 antibodies was 12.99% (95%CI, 7.21 - 22.28) at baseline and 24.10% (95%CI, 16 .17 - 34.31) by the end of the study. Persistence of anti-SARS-CoV-2 antibodies among seropositive participants was at least 6 months. Low education level and family income ≤ 3 minimum wages were significantly associated with the presence of IgG anti-SARS-CoV-2 antibodies. Individuals who did not complete high school were 10.76 times (95%CI, 1.051 – 110.21) more likely to be seropositive. The presence of COVID-19-related signs and symptoms was also significantly associated with seroconversion. Among these, the most frequently reported were fever, sore throat, headache, dyspnea and loss of smell or taste. The results allow us to conclude that the investigated laboratory workers had a relatively high prevalence of IgG anti-SARS-CoV-2 antibodies, although no evidence of transmission of COVID-19 in the workplace was found. Biosafety education and non-pharmacological prevention measures were efficient in mitigating the transmission of COVID-19 among the team of professionals and collaborators of the extension laboratories of the Faculty of Pharmacy of Federal University of Bahia.

7
  • Islania Almeida Brandão Barbosa
  • THE IMPORTANCE OF THERAPEUTIC MONITORING IN THE OCCURRENCE OF SERIOUS TOXICITY TO THE CHEMOTHERAPEUTIC FLUROURACIL IN PATIENTS WITH GASTROINTESTINAL NEOPLASIA

     
    Ícone "Verificada pela comunidade"
     
  • Advisor : ANA LEONOR PARDO CAMPOS GODOY
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • VANESSA BERGMANIN BORALLI MARQUES
  • Data: Aug 19, 2022


  • Show Abstract
  • Introduction: Cancer is the main public health problem in Brazil and worldwide, with emphasis on gastrointestinal cancer, which has a major association with environmental factors and lifestyle habits. The drug 5-fluorouracil is one of the main treatment alternatives for gastrointestinal cancers. The extremely short half-life, narrow therapeutic index, and inter-individual differences among patients lead to a limitation in its use due to the high possibility of occurrence of toxicities. Objective: This observational, longitudinal, retrospective, descriptive, uncontrolled single-center study aimed to assess the occurrence and severity of toxicities presented by patients using 5-fluorouracil medication and the importance of dose optimization and individualization. Methodology: The identification of the severity of 5-FU toxicities was based on the Common Toxicity Criteria, from the Common Terminology Criteria for Adverse Events (CTCAE) – version 5.0. Parametric ANOVA, non-parametric Kruskal-Wallis, and Chi-square (χ2) tests were used, in addition to the Mann-Whitney tests, in the statistical analysis of the data. Results and Discussion: Eighty-three patients undergoing treatment for colorectal cancer who underwent treatment with the FOLFOX protocol were evaluated (oxaliplatin 85 mg/m² IV in 2h of infusion; folinic acid 400 mg/m² IV in 2h of infusion; 5-FU 400 mg /m² in bolus; 5-FU 2400 mg/m² IV in 46 to 48 hours of infusion), between June 2020 and July 2021. The analyzes performed showed a higher frequency in males of developing toxicity, with hematological toxicities being more frequent in both sexes, in addition to presenting with greater severity. In addition, 44.6% of patients did not complete the proposed treatment, with disease progression being the main cause. Interferences in treatment due to adverse events to 5-FU were reported 112 times, leading to delay, temporary discontinuation of treatment (with or without hospitalization), complete discontinuation, and/or dose adjustment. BMI and development of severe toxicities were significantly dependent (p = 0.000). Conclusion: The results suggest a strong association between the presented toxicities and the need to suspend treatment and/or dose adjustments with a possible reduction in therapeutic efficacy and increase in healthcare costs. This study demonstrates the need for individualized treatment with more adequate management of adverse events.

8
  • TAIANE CANDEIAS DA SILVA
  • THE CHALLENGES OF THE USE OF VENETOCLAX IN PATIENTS WITH ACUTE MYELOID LEUKEMIA.
  • Advisor : ANA LEONOR PARDO CAMPOS GODOY
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • FRANCINE JOHANSSON AZEREDO
  • IZABEL ALMEIDA ALVES
  • Data: Sep 8, 2022


  • Show Abstract
  • Introduction: Venetoclax is a selective inhibitor of lymphoma B cell (BCL-2), an anti-apoptotic protein. Was approved in 2018 by major regulatory agencies in the United States, European Union and Brazil for the treatment of patients with Acute Myeloid Leukemia (AML) that are refractory to previous treatments or with relapse. Innovations in cancer treatment have directed therapies to target cells. These therapies are more tolerable than conventional chemotherapies, however potential, drug interactions are more frequent. Knowledge of drug interactions, individualization treatment and adequate pharmacotherapy management are important to ensure therapeutic success, as it offers the patient less experience with adverse events, improved treatment adherence, reduced additional costs to manage drug-related problems, improves the patient's quality of life and has a smaller impact on costs to public coffers than those that are already spent directly with the provision of treatment. Objective: Describe drug interactions and the challenges of using venetoclax in patients with AML at a Sentinel Hospital in Salvador. Methodology: This descriptive, observational and retrospective study. Data collection was performed using secondary data obtained from the search of medical records and clinical documents of these patients. Drug interactions induced by treatment with venetoclax were identified using the website drugs.com, which classifies interactions according to the effect and necessary management in: contraindicated, major, intermediate and minor. For data evaluation, parametric and non-parametric statistical tests were used. Results and Discussion: Ten patients who used venetoclax were evaluated, 100% of the patients (n=10) had a diagnosis of AML. Mean age was 46 years with standard (SD=17). A total of 319 drug interactions were found in the 10 patients and found that the greater the number of drugs prescribed, the greater the number of interactions. Drug interactions with venetoclax were identified as serious and contraindicated. About drug interactions, with venetoclax, the most frequent were venetoclax + voriconazole, followed by venetoclax + morphine, which are important drugs used in oncology. When it comes to contraindicated drug interactions, regarding the interactions mechanism, it was observed that 40% are pharmacokinetic, however, the serious interactions with venetoclax 100% are pharmacokinetic mechanisms involving the enzymes of Cytocromop450 and P-glycoprotein Conclusion: Drug interactions related to CYP and Glycoprotein P are the main limiting factors for the use of venetoclax in combination to other drugs. Therefore, a monitoring system needs to be adopted to check and evaluate these interactions and access their efficacy and safety, as much as the adverse arising events and impacts happened in cancer treatment. These impacts can be lead to an increase in the length of hospital stay or be fatal

9
  • ELDER TRINDADE DAMASCENO
  • EVALUATION OF HIGH-DENSITY LIPOPROTEIN IN PATIENTS WITH CHRONIC KIDNEY DISEASE IN REPLACEMENT THERAPY: BEFORE AND AFTER KIDNEY TRANSPLANTATION
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • CYNARA GOMES BARBOSA
  • JOSE PEREIRA DE MOURA NETO
  • RICARDO DAVID COUTO
  • Data: Nov 23, 2022


  • Show Abstract
  • Chronic kidney disease (CKD) is a worldwide relevant public health problem, with early cardiovascular disease (CVD) as the most cause of death. Disorders in lipoprotein metabolism are associated with CVD increasing, which makes it imperative to advance in lipoprotein structure and functionality studies, especially for high-density lipoprotein (HDL), as it acts in the reverse cholesterol transport and has cardioprotective, antioxidant, and anti-inflammatory properties, very important aspects in patients with CKD. This study aimed to evaluate HDL structural modifications in patients with CKD undergoing renal replacement therapy (RRT), hemodialysis, before and after kidney transplantation. This study included 50 patients with CKD undergoing hemodialysis before and after four months of kidney transplantation. Biochemical analyses were performed by reflectance, HDL particle size, and polydispersity by the light scattering method (laser light scattering) and the Apo-AI and Apo-B by turbidimetry. Significant increases were found among total cholesterol, non-HDL cholesterol, and LDL-c concentrations after transplantation. For Apo-B and the ApoB/Apo A-I ratio, we observed a significant increase after transplantation only in men. C-reactive protein was significantly lower after transplantation. No significant differences were found in HDL particle size before and after kidney transplantation. However, when analyzing the HDL particles polydispersities, the dispersion was lower after transplantation. The study findings showed significant differences in lipid metabolism before and after kidney transplantation. To properly assess the findings and their possible influence as a condition that increases cardiovascular risk, it will be necessary to develop more in-depth HDL structure and functionality studies, mainly considering the type of study design to reach the appropriate goals intended to be evaluated.

     

10
  • FABIANE MARIA FERNANDES NEVES
  • Development and validation of an analytical method using HPLC for the quality control of tenofovir disoproxil fumarate and emtricitabine.

  • Advisor : EDITH CRISTINA LAIGNIER CAZEDEY
  • COMMITTEE MEMBERS :
  • MARTIN STEPPE
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Nov 25, 2022


  • Show Abstract
  • Pre-exposure prophylaxis (PrEP) to the HIV virus (Human Immunodeficiency Virus - HIV) with the use of Truvada, consists of the combination of the fixed-dose oral formulation of the drugs emtricitabine (FTC) 200 mg and tenofovir disoproxil fumarate (TDF) 300 mg. This antiretroviral drug, whose main objective is to prevent the virus from seroconverting in the human organism, was approved by the Food Drug Administration (FDA) and made available in Brazil in 2017 by the Brazilian Unified Health System. However, analytical methodologies developed for the quality assurance of Active Pharmaceutical Ingredients (API) of Truvada are limited. The purpose of the present theoretical-experimental study was to analyze and elaborate a literature review of existing analytical and bioanalytical methods in the literature for the antiretroviral drugs FTC and TDF. In addition, in the experimental phase, a rapid isocratic reverse-phase analytical method was developed and validated by High-Performance Liquid Chromatography (HPLC) with simultaneous UV detection of the active drugs. The study demonstrated adequate validation parameters among sensitivity, linearity, precision, accuracy, and limits of detection and quantification, meeting the main national and international regulatory guides. Therefore, the method proved to be suitable for use in routine analysis for active pharmaceutical ingredients or tablets.

     


11
  • Laiz Campos Pereira
  • Amphotericin B Pharmacokinetic/pharmacodynamic modeling against Candida Albicans

  • Advisor : FRANCINE JOHANSSON AZEREDO
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • BIBIANA VERLINDO DE ARAUJO
  • FRANCINE JOHANSSON AZEREDO
  • Data: Dec 14, 2022


  • Show Abstract
  • Invasive fungal infections (IFIs) are considered a public health problem, and recent studies have shown that the Candida albicans is responsible for most cases. For the treatment of this type of infection, antifungal drugs that belong to the classes of azoles, polyenes, and echinocandins are used, but factors such as fungal resistance, adverse effects related to toxicity, and interactions between drugs difficult the therapeutic success of the treatment. Amphotericin B (AmB), the only representative of the polyene class, presents a broad spectrum of antifungal action and lower resistance rates, with AmB associated with sodium deoxycholate (AmB), considered the conventional formulation, the most used in Brazilian public hospitals. In order for the pharmacological treatment to be successful, it is necessary to have an in-depth knowledge of both the pharmacokinetic (PK) and the pharmacodynamic (PD) aspects of the drug, and the pharmacokinetic-pharmacodynamic (PK/PD) modeling is considered the most effective approach to assess the effectiveness of antifungal agentes. Therefore, this work aimed to develop a PK/PD modeling of AmB deoxycholate against C. albicans. This work consists two chapters: (i) review article about main approaches of the PKPD modeling and studies applying this approache with antibacterials and antifungals; (ii) PK/PD modeling of antifungal effect of AmB-D agains C. albicans, by time-kill curves obtained fram na in vitro model, with satic and dynamic concentrations. For the PK/PD modeling, it was be an adapted sigmoidal Emax model by software Monolix ®. From the obtained parameters (Emax and EC50), it was possible to assess the potency and effectiveness of AmB Against C. albicans, and comparing the results from two different methodologies.

12
  • Miguel de Jesus Oliveira Santos
  • EVALUATION OF MICROALGAE IN OBTAINING PHARMACEUTICAL PRODUCTS AND TREATMENT OF THEIR WASTE

  • Advisor : HENRIQUE RODRIGUES MARCELINO
  • COMMITTEE MEMBERS :
  • SILVIA STANISCUASKI GUTERRES
  • HENRIQUE RODRIGUES MARCELINO
  • NEILA DE PAULA PEREIRA
  • Data: Dec 15, 2022


  • Show Abstract
  • By introducing the concept of biorefinery for microalgae, a circular economy system can be envisaged in the pharmaceutical industry. This system is based on the extraction of active pharmaceutical ingredients and excipients and their bioremediation in wastewater and effluents containing the same components as well as other residues. Therefore, the aim of this work is to analyze the feasibility of microalgae biorefineries in the pharmaceutical industry on two fronts. Thus, a literature review on the bioremediation of drug contaminated media by microalgae is presented in this work. In addition, an extract of the protein fraction of the microalgae Arthrospira sp. LEB-18 was obtained for further investigation. During the literature review, it was found that microalgal systems are similarly efficient as conventional treatment methods in removing drugs from water. The growth of microalgae exposed to the drug does not appear to be affected at realistic concentrations. The main removal mechanism is biodegradation. In the experiments, Arthrospira platensis LEB-18 was microencapsulated in Eudragit® L100 at a 2full factorial design. The entrapment efficiency was related to the larger polymer to biomass ratio. No changes in the chemical profiles of the raw materials were observed in the microparticles, which also exhibited a semi-crystalline profile. Microscopic images showed microparticles but also larger biomass fragments that were not fully encapsulated. In vitro digestibility tests indicated protection of the proteinaceous fraction of the encapsulated biomass. The feasibility of a pharmaceutical biorefinery could be achieved in the short to medium term if technological breakthroughs in research are confirmed.

2021
Dissertations
1
  • EDUARDO MOREIRA DA SILVA
  • EVALUATION OF THE IMPACT OF INTERVENTIONS IN A PUBLIC CLINICAL LABORATORY AFTER QUALITY MANAGEMENT AND STRATEGIC PLANNING
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • RICARDO DAVID COUTO
  • LUCIANA SANTOS CARDOSO
  • FERNANDA WASHINGTON DE MENDONCA LIMA
  • Data: Jan 21, 2021


  • Show Abstract
  • In Brazil, in the last three decades, quality management has entered the scenario of laboratory services for clinical analysis. This scenario showed up by the growing number of certifications and accreditations regulated by the INMETRO, mainly in private equity organizations. However, there is still limited interest in the implementation of quality management by public institutions linked to SUS. This dissertation demonstrates the importance of quality management and strategic planning for improving quality concerning products, services, process gear, information flows, organizational performance, medical decision making, and risk analysis within these public institutions. To carry out this exploratory-descriptive study on the Laboratory of Clinical and Toxicological Analysis (LACTFAR) from the Faculty of Pharmacy, Federal University of Bahia (UFBA), techniques, tools, and methodologies for quality management were used to achieve the results. In this sense, non-conformities could be shared into categories to emphasize infrastructure and process, which accounted for 78.1%, while the operational process accounted for 21.9% of occurrences in the general non-compliance index. This find indicates the direction that should be focused within the LACTFAR structure for the preparation of action plans and the application of resources for solving problems. Thus, we found that the process approach through process control is a fundamental tool because such results showed the non-conformities that were previously treated as ordinary situations and with no apparent solution. The knowledge of the interrelationships of the processes favors the identification of critical points to establish constant monitoring that provides visible benefits in the quality of the final product, thus reducing the rework shown in the reduction of non-conformities. Organizational development occurs based on internal changes guided by looking at changes in external scenarios, understanding the context in which the institution is inserted for its strategic direction.

2
  • GESSICA SABRINA DE ASSIS SILVA
  • THERAPEUTIC POTENTIAL OF EXTRACELLULAR VESICLES DERIVED FROM MESENQUIMAL CELLS IN DIABETIC SENSORY NEUROPATHY IN MICE
  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CRISTIANE FLORA VILLARREAL
  • ANA LEONOR PARDO CAMPOS GODOY
  • LUIZ FERNANDO FERRARI
  • Data: Feb 1, 2021


  • Show Abstract
  • INTRODUCTION: Sensory neuropathy is one of the most frequent and debilitating complications of diabetes and manifests itself through paradoxical symptoms such as loss of sensation. Its pharmacological treatment is not very effective, produces a series of adverse effects and has no curative action, acting only in the relief of symptoms. Mesenchymal stem cells have been considered promising for the treatment of painful neuropathies. The therapeutic properties of acquired mesenchymal cells are attributed to their ability to release soluble bioactive molecules and contained in extracellular vesicles (EV). OBJECTIVE: To investigate the therapeutic potential of LV derived from human bone marrow mesenchymal cells (BMMCs) in a murine model of sensory diabetic neuropathy (SDN). METHODS: BMMCs were slid from the human bone marrow and characterized by morphological analysis, flow cytometry and in vitro differentiation. The EV were adjusted by ultracentrifugation of the BMMCs secretome and characterized by their size, shape and quantity by transmission electron microscopy (TEM) and by the suspended particle analysis technique (NTA - Nanoparticle Tracking Analysis). The streptozotocin NDS model (ETZ; 80 mg / kg / ip) was induced in male C57Bl / 6 mice (22-25g), from the IGM Biotery (FIOCRUZ-BA) and the nociceptive threshold was performed throughout the period . experimental (92 days), with von Frey filaments. Thirty days after the model was induced, the mice received intravenous administration of saline (100 µL), EV-BMMCs (100 µL) or BMMCs (1x106). The contribution of the opioid system to the analgesic effect of cell therapy was investigated in an antagonist reversal assay, using naloxone (0.4 mL / kg i.p.) administered 7 and 60 days after transplantation. An expression of preproencephalin in the spinal cord and no supernatant from the BMMCs culture was analyzed by quantitative Real-Time PCR (RT-qPCR). RESULTS: Through the differentiation assay and flow cytometry characterization, it was confirmed that the expanding cells were mesenchymal. The characterization of EV-BMMCs by NTA describes an EV population with a size of 52.4 to 450 nm, corresponding to microvesicles and exosomes. The average total number of vesicles recovered from 1x106 BMMCs was 4.4x108. TEM data confirmed the presence of structures with morphology similar to that of EV. A single intravenous administration of BMMCs or EV-BMMCs was able to reverse the behavioral signs of painful diabetic neuropathy throughout the experimental period (p <0.05). The administration of naloxone (3 mg / kg, i.p.), a non-selective opioid receptor antagonist, reversed an antinociception induced by BMMCs and EV-BMMCs (p <0.05). Analyzes of RT-qPCR performed 7 days after treatments with BMMCs and EV-BMMCs indicate an increase in the expression of preproencephalin in neuropathic animals when compared to saline (p <0.001). Already 60 days after the treatments, it was observed that neuropathic animals treated with less saline expression of preproencephalin in the spinal cord compared to non-neuropathic animals (p <0.05). In addition, the administration of EV-BMMCs, but not BMMCs, increased the expression of preproencephalin in the spinal cord of mice with diabetic neuropathy when compared to neuropathies treated with saline (p <0.01). In addition, analyzes of BMMCs in culture showed that these cells express preprooencephalin. CONCLUSION: the results of the present study show that EV derived from human EV-BMMCs preserve the antinociceptive properties of the cells of origin, and that this effect seems to involve the activation of the endogenous opioid system. This study highlights the potential of cell-free therapy to control painful diabetic neuropathy.

3
  • JUCELINO NERY DA CONCEIÇÃO FILHO
  • Evaluation of Baseline Levels of Butyrylcholinesterase in a Sample of the Population of Salvador and the Epidemiological Profile of Exposures to Pesticides in Bahia (2017 to 2019)

  • Advisor : ANA LEONOR PARDO CAMPOS GODOY
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • CLAUDIA REGINA DOS SANTOS
  • MARILDA DE SOUZA GONCALVES
  • Data: Feb 19, 2021


  • Show Abstract
  • INTRODUCTION: Exogenous intoxications are an important worldwide public health problem, with significant challenges for managers in this area, with pesticides as one of the main causal agents. Knowledge of the epidemiological profile of this disease in the state of Bahia is an important tool for decision making by public health agencies, therefore, information systems need to have quality and reliable data. In the field of laboratory diagnosis, Butyrylcholinesterase (BChE) is a relevant biological indicator in the diagnosis and monitoring of these intoxications. OBJECTIVES: The present study aims to investigate the baseline values of BChE activity in a sample of the population of Salvador, taking into account the factors that may interfere with enzyme activity, and to characterize the profile of cases of exposure to pesticides in the state of Bahia in the period from 2017 to 2019, identifying the frequency of intoxications involving cholinesterase inhibiting substances. METHODS: The present study consists of two stages. The first consists of a descriptive, quantitative study, with a transversal approach. The sample consisted of blood donor volunteers in the state's blood center. The second step is a descriptive observational cross-sectional study, considering the aggregate of data related to exogenous intoxications registered in Bahia in the period from 2017 to 2019, using secondary data and a quantitative approach. RESULTS: The present study validated the methodology used by the Toxicological Analysis Laboratory (Labtox) of the Bahia Toxicological Information and Assistance Center (CIATox-BA), in addition to making it possible to know the baseline levels of butyrylcholinesterase in a sample of the population of Salvador, which has shown activity between 6,000 to 20,500 U / L, with a lower average in females. The values obtained for males were 9,300 U / L to 18,400 U / L and 8,200 U / L to 18,800 U / L for females. Regarding epidemiological data, this study made it possible to identify the occurrence of inconsistencies and incompleteness in both information systems, in addition to indicating an underreporting in Sinan of 46.7%, despite the compulsory notification of the disease studied. There were 2,853 pesticide poisonings, 51.0% of which were male and 48.7% female. The occurrences predominated in homes (59.7%) and suicide attempt was the main circumstance (48.4%). The lethality found was 2.4%, being higher in males. The most frequent active ingredients were cholinesterase inhibiting insecticides. CONCLUSION: The results obtained here make it clear that, despite underreporting, pesticide poisoning is not limited to rural areas, crops, men or adults, requiring broader action by public authorities, also evidencing the need to better qualify the information systems feed. In addition, the high frequency of cholinesterase inhibitors signals that BChE still has its important role as a biological indicator in exposure to pesticides, regardless of the exposure environment.

4
  • JONATHAM SOUZA MOREIRA
  • EVALUATION OF A SERIES OF THYOSEMICARBAZONES AS INHIBITORS OF NEW DHELI METALLO-BETA-LACTAMASE-1

     



  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • FRANCO HENRIQUE ANDRADE LEITE
  • JOICE NEVES REIS PEDREIRA
  • Data: Mar 8, 2021


  • Show Abstract
  • In recent years, the global spread of bacterial resistance to β-lactam drugs has been observed, with carbapenem derivatives having an important role in the treatment of infections by multi-resistant bacteria. The expression of β-lactamase is mainly related to bacterial resistance, which requires the development of bacterial resistance blockers. Although combinations of β-lactam drugs and serine-β-lactamase inhibitors have been successful, such inhibitors are inactive against Metalo-β-lactamases especially, New Delhi Metalo-β-lactamase (NDM). So far, few compounds are active against NDM-producing bacteria and no specific inhibitors are available. A rational development strategy for NDM inhibitors begins with in vitro assays with the aim of identifying compounds that can act synergistically with β-lactam antibiotics. Thus, thiosemicarbazone derivatives were synthesized and investigated for their ability to reverse the NDM-resistant phenotype in Enterobacter cloacae. Phenotypic screening indicated that four beta-isatin-thiosemicarbazones showed Fractional Inhibitory Concentration (FIC) ≤ 250 µM in the presence of Meropenem (2 µg/mL) with compound 17, the most promising, (FIC = 31.25 µM) showing an effect synergistic (FIC index = 0.34). Docking and Molecular Dynamics studies on NDM-compound 17 complex suggested that 2,3-dihydro-1H-indol-2-one subunit of isatin-β interacts with catalytic zinc and performs hydrogen bonds with Asp-124 acting as a non-classical bioisostere of carboxylic acid. In addition, the tautomeric state of thiosemicarbazone with oxidized sulfur appears to act as a spacer instead of a zinc chelator and the aromatic portions are stabilized by pi-pi and pi-cation interactions with His189 and Lys221 residues. Our results addressed some structural characteristics of thiosemicarbazone in complex with NDM, and highlights its scaffold as promising alternatives to face bacterial resistance.

5
  • GUSTAVO REIS SAMPAIO
  • CONSUMPTION OF PSYCHOACTIVE SUBSTANCES AMONG STUDENTS IN THE HEALTH AND BIOLOGICAL SCIENCES OF UFBA
  • Advisor : DENIS DE MELO SOARES
  • COMMITTEE MEMBERS :
  • FABIO CARDOSO CRUZ
  • DENIS DE MELO SOARES
  • DJANILSON BARBOSA DOS SANTOS
  • Data: Nov 18, 2021


  • Show Abstract
  • Psychoactive substances act on the central nervous system, producing changes in sensations, in the degree of consciousness or in the emotional state, whether intentionally or not. The consumption of psychoactive substances is not a recent phenomenon. Since prehistory these substances have been present between different cultures. Three patterns of drug use can be identified: 1) occasional or controlled use; 2) abusive or harmful use; 3) compulsive use or addiction. Some epidemiological studies have been carried out in Brazil in order to verify the prevalence of drug use among the university population. Most of them agree that the use of alcohol and other substances is higher among university students when compared to the general population and high school students. Regarding the consequences of the consumption of psychoactive substances, among university students, we could cite: car accidents, violence, risky sexual behavior, low academic performance, decreased perception and stress. The university students of health courses deserve a special mention when it comes to the use of psychotropic drugs, as they will be responsible for the health education of the population in the future, responsible for identifying and referring patients with problems related to the use of psychotropic drugs and also because they serve as a model for their patients. Therefore, this paper aims to describe the substance consumption profile by students in Area II (Physiotherapy, Biological Sciences, Nutrition, Pharmacy, Nursing, Biotechnology, Public Health, Animal Science, Dentistry, Veterinary Medicine, Medicine, Natural Sciences, Gastronomy and Speech–Language Pathology) at UFBA, through the application of an online questionnaire. Most of the questions were taken from ASSIST (WHO’s Alcohol, Smoking and Substance Involvement Screening Test) and other works related to this topic. The data were processed using the SPSS statistical program. 514 university students have answered the questionnaire, of which 502 have been assessed according to the inclusion criteria. Among the participants, the mean age was 25.2 years with a standard deviation of 7.7 years, 72.9% were female and 27.1% were male. For the use of prescription psychoactive drugs, 12.9% of the participants revealed their use at least at some point. For the use of psychoactive substances in a recreational way, both for use in life and for use in the last 3 months, the prevalence of alcohol, marijuana and tobacco were the highest. According to the ASSIST questionnaire, alcohol was the substance that showed the highest proportion of individuals in need of more intensive treatment. The main reasons cited for the use of psychoactive substances were fun and controlling anxiety. The consumption profile varied in relation to the literature, where marijuana appeared as the second most consumed substance. The present paper managed to achieve the proposed objectives and not only confirmed some data observed in other studies carried out with university students, but also brought new information about the extent and profile of consumption of these substances.

6
  • KATHARINE VALÉRIA SARAIVA HODEL
  • DEVELOPMENT AND CHARACTERIZATION OF BASE BIOCURATIVES OF BACTERIAL CELLULOSE INCORPORATED WITH P-CUMARIC ACID, BIOCHANIN A AND GREEN AND RED PROPOLIS EXTRACT
  • Advisor : ANA LEONOR PARDO CAMPOS GODOY
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • CAROLINA OLIVEIRA DE SOUZA
  • TANIA FRAGA BARROS
  • Data: Nov 29, 2021


  • Show Abstract
  • Bacterial cellulose (BC) is one of the great bets of the biomedical sector because of its unique properties that are essential for an ideal dressing. Among them, the possible incorporation of molecules that have an active action in the healing process. Propolis and its biocomponents (biomarkers) are considered important candidates for incorporation in BC membranes for application as dressings, since they have important biological activities for tissue regeneration. In this context, the aim of this study was to develop and characterize BC-based biocuratives incorporated with green and red propolis extracts, as well as the active compounds p-coumaric acid and biochanin A. In total, nine biocuratives were developed (one control and eight active biocuratives) that were characterized for morphological, optical, physical and barrier, tensile mechanical, antioxidant and antimicrobial properties. The in vitro permeation using Franz diffusion cells of the biocuratives formed by CB and the active substances was also performed. Within this context, the characterization of the nine samples developed, where the transparency (28.59-110.62 T600.mm-1 ), opacity (28.43-50.48 Abs500.mm-1 ), weight (0.004-0.009 g.cm-2 ), thickness (0.023-0.046 mm), aw (0.292-0.455), water solubility (6.25-65.58 %), moisture content index (27.44-84.87 %), intumescence index (48.93-405.55 %), water vapor permeability rate (7.86-38.11 g.m².day-1 ), water vapor permeability (2.29-10.30 g-mm/m²-day-1), maximum stress (0.53-4.15 MPa), elongation (99.13-262.39 %), total flavonoid content (0.05-39.16 mg EQ.g-1 ), DPPH radical scavenging capacity (21.23-86.76 µg.mL1 ) were determined. The biocuratives based on BC and red propolis extract were the only ones that showed antimicrobial activity against Escherichia coli. The permeation and retention assay showed that the biocuratives containing propolis extract showed a higher concentration in the stratum corneum when compared to viable skin. The results showed, in general, that the presence and concentration of the active compound in the cellulosic matrix interferes in the characterization tests performed. Thus, the different performances as to the properties evaluated suggest that they have a potential application for different types of skin lesions, as well as for different phases of healing.

7
  • SAMARA ALVES SANTOS
  • Evaluation of Indirect Immunofluorescence and Western blotting in the detection of serum anti-Giardia duodenalis antibodies

  • Advisor : MARCIA CRISTINA AQUINO TEIXEIRA
  • COMMITTEE MEMBERS :
  • LUCIANA SANTOS CARDOSO
  • MARCIA CRISTINA AQUINO TEIXEIRA
  • THIAGO CAMPANHARO BAHIENSE
  • Data: Dec 2, 2021


  • Show Abstract
  • Giardia duodenalisis considered one of the oldest human parasites with wide distribution worldwide, infecting more than 280 million people a year. Giardiasis is a highly prevalent protozoosis in Brazilian children, with asymptomatic or acute or persistent diarrhea, accompanied by abdominal cramps, flatulence, malabsorption and weight loss. Laboratory diagnosis of this infection is usually performed through the morphological identification of cysts and/or trophozoites in the stool through microscopy. In addition, serological assays such as Indirect Immunofluorescence (IFI), ELISA (Enzyme-linked immunosorbent assay) and Western Blotting (WB) have been used to detect specific antibodies to Giardia in serum. However, there is much controversy regarding the use of these assays as diagnostic tools. Thus, this work aimed to evaluate the performance of Indirect Immunofluorescence (IFI) and Western blotting (WB) assays for the detection of IgG antibodies against Giardia duodenalis in infected individuals. Fecal and serum samples were selected from children from day care centers and their guardians, as well as from users of the Clinical Analysis Laboratory of the Faculty of Pharmacy (LACTFAR-UFBA) infected with Giardia duodenalis, infected with other parasites, or non-parasitized. Sera were analyzed using the IFI and WB technique in order to compare the sensitivity and specificity of the methods and identify immunodominant molecules of the parasite and possible cross-reactions with other parasites. Ninety-seven sera were tested in the IFI at 1:20 and 1:40 seric dilutions, and of these, 40 samples were tested in the WB assay. The sensitivity of the IFI was 97% at the 1:20 dilution of the sera, dropping to 39.4% at the 1:40 dilution. For the WB, the sensitivity was 85.7%, considering the total of 14 sera from patients eliminating Giardia cysts. However, the specificity for IFI ranged from 46.9% to 59.4%, depending on the serum dilution, while in WB it was 75%. Most of the serum samples from patients with amoebas were positive in the IFI at a 1:20 dilution, evidencing a possible cross-reaction, also observed in the WB. Proteins of 25, 27-31 and 45-55kDa were considered immunodominant because they were the most recognized by sera of people infected with G. duodenalis, presenting an exclusive diagnostic sensitivity of each protein of 42.8%, 50% and 57.1% respectively. The agreement between WB and the detection of G. duodenalis in feces by microscopy was considered substantial, Kappa = 0.61, while for IFI the serum dilutions 1:20 and 1:40 were considered low, Kappa = 0, 27 and 0.31, respectively. The agreement between the two serological methods tested was weak with sera diluted 1:20 in the IFI, Kappa = 0.150, or null, with a 1:40 dilution of sera. Constant lifetime exposure to Giardia infection can maintain high levels of specific antibodies in the serum, even without active infection, which may explain the high sensitivity and low specificity of assays. The use of fractionated antigens, as in WB, allows the recognition of specific immunodominant antigens and their combinations or exclusion criteria, which can be used as an alternative test in the diagnosis of giardiasis.

8
  • CEZAR MIGUEL SANTOS JUNIOR
  • Isolation and characterization of coumarins obtained from the stem bark of Metrodorea mollis
  • Advisor : RENATA BIEGELMEYER DA SILVA RAMBO
  • COMMITTEE MEMBERS :
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • JORGE MAURICIO DAVID
  • ALEXSANDRO BRANCO
  • Data: Dec 7, 2021


  • Show Abstract
  • The brazilian flora has around 50.000 identified species, wich means 22% of the existing vegetation worlwide. However, there are also many species without proper identification, specially the ones from northeastern region of Brazil. The genus Metrodorea (Rutaceae) comprises 6 species ranging between shrubs and trees, which are found exclusively in South America, and which belongs to the vegetation found in Chapada Diamantina National Park. Among the species found in Metrodorea, M. mollis has, so far, just one report regarding its phytochemical composition. However, there was no information about the part of the plant used for compounds isolation. In this contexto, this study aimed to fractionate the crude extracts obtained from stem barks of M. mollis, in order to provide accurate information about obtaining active compounds from this plant. Stem barks from M.mollis were collected in March 2005 in the district of Catolés, located in Abaira city, Chapada Diamantina-Bahia, Brazil. The plant was identified and deposited in Alexandre Leal Costa Herbarium by the voucher number 69153. Air-dried stem barks of M.mollis were macerated with hexane and methanol in order to obtain its corresponding extracts, which were fractionated by column chromatography and eluted with ethyl acetate and hexane and/or chloroform in gradient system. NMR 1H and 13C analyzes provided the identification of six coumarins: braylin (1), isopimpinellin (2), xanthoxotin (3), xanthoxyletin (4), alloxanthoxyletin (5), luvagentin (6). This study is the first to bring information about isolation and characterization of compounds (5) and (6) among all the Metrodorea genra.

9
  • ANANDA FREITAS FONSECA
  • DEVELOPMENT OF THE POPULATION PHARMACOKINETIC MODEL OF BUPROPIONA USING LITERATURE DATA
  • Advisor : ANA LEONOR PARDO CAMPOS GODOY
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • RICARDO DAVID COUTO
  • SANDRA ELISA HAAS
  • Data: Dec 14, 2021


  • Show Abstract
  • Introduction: Depression is one of the most common psychiatric disorders, due to the high incidence in this decade, a greater prevalence in women, including major depressive disorder in pregnancy. When the diagnosis is made, the treatment occur according to the degree of the disease and when this degree is moderate to severe, pharmacological treatment with antidepressants is started. Among them, the most prescribed drugs are selective serotonin reuptake inhibitors, they are taken first-line drugs because they are safer than the others. Bupropion is reported in the literature as a second-generation SSRI antidepressant, which has three different release systems: immediate (IR), sustained (SR) and extended (XL). Objective: The objective of this study is to develop a population pharmacokinetic model using literature data to assess the pharmacokinetic profile of bupropion in its three delivery systems (IR, SR and XL) in addition to analyzing the influence of the covariates sex and smoking. Methodology: For this, an exploratory search was carried out with well-established inclusion, non-inclusion and exclusion criteria in order to find articles that had performed pharmacokinetic studies in human subjects and administered bupropion in any of its delivery and dose systems. Another essential feature of these studies is to contain at least one graph of concentration versus time with sufficient definition for their data to be extracted using the WebPlot Digitizer software. After getting the data, the population pharmacokinetic model was developed using the Monolix® Suite 2020R1 software. Results: After applying the research strategies, it was reached an number of 5 articles that corresponded to the characteristics necessary to perform the data extraction and the scope of the population pharmacokinetic model that harmoniously adjusts and explains the studied covariates. As a result, we obtained a one-compartment model, with zero-order absorption, linear elimination and considering the lag time related to the different release systems. Furthermore, we could see that the covariates sex and smoking do not influence the pharmacokinetics of bupropion, as found in other studies. Conclusion: The population pharmacokinetic model developed from information extracted from studies in the literature was able to adapt the data from these studies and explore the proposed covariates. As expected, the type of delivery system has a strong interaction with drug release delay time. On the other hand, the covariates sex and smoking do not influence the pharmacokinetic parameters of bupropion.

10
  • AISLA MÉRCIA LÁZARO DE OLIVEIRA
  • MOLECULAR CHARACTERIZATION OF INVASIVE ISOLATES OF Streptococcus pneumoniae IDENTIFIED IN THE POST-VACCINE PERIOD

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • JOICE NEVES REIS PEDREIRA
  • MARCIA CRISTINA AQUINO TEIXEIRA
  • CÍCERO ARMÍDIO GOMES DIAS
  • Data: Dec 17, 2021


  • Show Abstract
  • Streptococcus pneumoniae or pneumococcus, a gram-positive bacterium with 97 serotypes described, is one of the main causes of invasive infections that affect mainly children and the elderly, even though it is preventable by vaccines with good efficacy. In Brazil, the PCV10 conjugate vaccine that includes serotypes 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F, and 23F is available in the national vaccination plan, which contributed to a reduction of more than 90% of cases of invasive infections caused by pneumococci. This study aimed to characterize the phenotypic and genotypic profiles of S. pneumoniae isolates from cases of invasive disease from 2010 to 2019 in
    Salvador, Brazil. The isolates were obtained through the microbiological culture of different clinical specimens. The serotype was determined by multiplex PCR, the antimicrobial susceptibility profile was determined by disk diffusion method, and the clonal profile was obtained by PFGE and MLST. During the study period, we identify 226 cases of pneumococcal infection. Isolates were obtained from CSF (46.5%), blood (38.9%), CSF and blood (10.6%), pleural fluid (3.0%), tracheal aspirate (0.4%), and bronchoalveolar lavage (0.4%). When comparing the years 2010 and 2019, there was a reduction of 90.2% of cases of invasive pneumococcal disease (p=0.02). Most cases were due to non-vaccine serotypes, 67.3% (152/226), and the remaining were affected by vaccine serotypes, 32.7% (74/226). The most frequent serotypes were 14 (9.3%), 3 (8.8%), 23F (7.5%), 12F (6.2%) 19A (4.9%), 4 (4.4%), and 8 (4.4%). Male patients were more frequent (58.8%). The median age was 36 years [9-54, interquartile range 25-75%). Children aged 0-2 years (p<0.001) and 3-4 years (p=0.03) were more affected by pneumococcal infection by vaccine serotypes (14, 18C, 23F, and 6B), while adults aged 25-59 years were more affected by non-vaccine serotypes (12F, 19A, 3, and 8) (p=0.048). The antimicrobial susceptibility test showed that, of the included isolates, 23.9% (54/226) were not susceptible to penicillin, 22.1% (50/226) were not susceptible to sulfamethoxazole/trimethoprim, and 19.5% (44/226) were not susceptible to tetracycline. The vaccine serotypes 14, 19F, 23F, 18C were more resistant to penicillin (p>0.001) and to sulfamethoxazole/trimethoprim (p>0.001), while the non-vaccine serotypes 15C, 31, 19A were more resistant to erythromycin (p=0.02). Multidrug resistance was observed in 6.2% (14/226) of the isolates, and of these, 42.9% (6/14) were serotype 19A. Regarding the molecular analysis, three main clonal groups were observed, formed by ST66 (Serotype 14), ST218 (Serotype 12F), and ST193 (Serotype 18C). These findings reinforce the importance of continuous surveillance in cases of invasive disease caused by S. pneumoniae so that new epidemiological data are provided and help in evaluating the impact of the PCV10 vaccine in Brazil.

11
  • CLEVERSON ALVES FONSECA
  • MOLECULAR AND INFLAMMATORY CHANGES ASSOCIATED WITH ANGIOGENESIS IN
    MULTIPLE MYELOMA PATIENTS FROM SALVADOR BAHIA

  • Advisor : MARILDA DE SOUZA GONCALVES
  • COMMITTEE MEMBERS :
  • CYNARA GOMES BARBOSA
  • MARILDA DE SOUZA GONCALVES
  • MARINHO MARQUES DA SILVA NETO
  • Data: Dec 27, 2021


  • Show Abstract
  • Multiple Myeloma (MM) is a hematologic neoplasm characterized by clonal proliferation of plasma cells in the bone marrow (BM), which alter the marrow microenvironment mediated by inflammatory cytokines and angiogenic factors, representing 1% of all neoplasms and 13% to 20% of hematologic neoplasms. Its main clinical manifestations are associated with lesions in several organs, such as the kidneys, bones, and OM, with evolution to hypercalcemia, renal failure, osteolytic lesions, anemia, in addition to presenting an immunodeficiency picture. Thus, the present study investigated laboratory alterations associated with the stages of multiple myeloma, as well as the presence of polymorphisms in the IL-1B, IL-6, TNF-A, VEGF, and VDR genes and their associations with laboratory markers in patients with MM me SalvadorBA. Therefore, a cross-sectional study was carried out involving 22 individuals diagnosed with MM treated at the Hematology Service of CHUPES-UFBA and 55 healthy individuals, adjusted for age and gender, followed up at the LACTFAR-UFBA, after approval by the CEP. Analyzes to determine laboratory biomarkers were performed using an automated method, following the manufacturer's instructions; the molecular analyzes to determine the genetic polymorphisms were performed by gene sequencing, through the sequencing platform of the IGM-FIOCRUZ. The results showed a mean age of 55.1 ± 9.8 years in the MM group, with 63.64% (n=14) female; Furthermore, 75% (n=17) of these individuals are black and mixed-race, and 38.1% (n=8) were diagnosed with stage I disease (ISS). It was shown that individuals diagnosed with MM had increased levels of total protein and urea and reduced levels of albumin and phosphorus when compared to healthy controls. The C allele for IL-6 rs1800795 polymorphism was associated with the presence of MM. The association of genetic polymorphisms with laboratory markers in patients with MM demonstrated an association of the A allele of TNF-A rs1800629 with increased levels of total proteins and reduced levels of albumin; the A allele of TNF-A rs673 with increased serum calcium levels; the A allele for VEGF rs699947 with reduced serum phosphorus concentrations; IL-1B rs16944 A allele with elevated serum calcium concentrations; no associations between VDR rs2228570 polymorphism and laboratory markers were observed. Thus, the present work suggests the IL-6 rs1800795 polymorphism as a possible marker for the development of MM, demonstrating the importance of studies that evaluate genetic polymorphisms, mainly involving the expression of cytokines and angiogenic factors and their associations with biomarkers, as they may contribute to a better understanding of the mechanisms involved in the development and evolution of the MM.

2020
Dissertations
1
  • ALYSSON LUIZ MENDES DA SILVA
  • Determination of paraoxonase and arylesterase activities of PON1 enzyme on the visible spectrum range in patients with dyslipidemia: low HDL-c and hypertriglyceridemia

  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • RICARDO DAVID COUTO
  • ANA LEONOR PARDO CAMPOS GODOY
  • SIMONE GARCIA MACAMBIRA
  • Data: Feb 17, 2020


  • Show Abstract
  • Disorders in lipid and lipoprotein metabolism (dyslipidemia) are common in the population and are considered risk factors for cardiovascular disease. Studies have shown enhanced cardiovascular risk in individuals with PON1 low paraoxonase activity. However, the role of the arylesterase activity of PON1, an enzyme associated with HDL, is not yet known. This cross-sectional study aimed to evaluate arylesterase/paraoxonase activity of PON1 in serum samples from normolipidemic patients and with dyslipidemia such as hypertriglyceridemia and low HDL-c. Sixty serum were evaluated and stratified in three groups. Lipid profiles were determined, apoA1 levels measured and paraoxonase/arylesterase enzymatic activity of PON1 was determined against paraoxon and phenylacetate substrates, respectively, by spectrophotometry in kinetic mode at 405nm using 96 well microplates. The assays showed a significant difference in enzyme activity between the different groups studied and a proportional reduction in PON1 enzymatic activity compared to normolipidemic patients. After the correction of enzymatic activities by apoA1 concentration, the low HDL-c group showed the largest reduction in enzymatic activity and positive linear correlation when evaluated the concentration of HDL-c and apoA1. There was also a directly proportional dependence between enzymatic activity and apoA1 levels variables under normolipidemic and low HDL-c conditions after linear regression. The results suggest that for hypertriglyceridemia conditions, there is a decreased activity to a lesser extent when compared to the condition of low HDL-c, where is observed a significant reduction in enzymatic activity of PON1.

2
  • ADRIELE PINHEIRO BOMFIM
  • CHARACTERIZATION OF KLEBSIELLA PNEUMONIAE ISOLATES FROM BLOOD CURRENT INFECTIONS

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • JOICE NEVES REIS PEDREIRA
  • JOSILENE BORGES TORRES LIMA MATOS
  • TANIA FRAGA BARROS
  • Data: Mar 9, 2020


  • Show Abstract
  • Klebsiella pneumoniae is an important pathogen related to bloodstream infections (BSI), and a public health problem, mainly due to the emergence of strains resistant to multiple antimicrobials and the spread of mobile genetic elements encoding β-lactamases. This study aimed to describe the clinical and epidemiological aspects of BSI caused by K. pneumoniae as well as to determine the phenotypic and genotypic profile of antimicrobial resistance. This is a cross-sectional study conducted in two private tertiary hospitals, from April/2016 to December/2018, in which all cases of ICS by K. pneumoniae were evaluated. Epidemiological and clinical data were obtained by reviewing medical records, phenotypic resistance profiles were determined using the automated system VITEK-2® (bioMeriéux), the β-lactam resistance genes were determined by PCR, and the genetic profile by PFGE. A total of 196 cases of BSI due to K. pneumoniae were identified, 87.7% were classified as healthcare-related infections and the patients had a median age of 67 years (55-80) and the majority were male (59.2%). Resistance to carbapenems was identified in 31% of cases and resistance to cephaloporins in 64.4%. Younger patients (median 60 years old) and with kidney disease were at higher risk for KP-CR infection. The overall death rate was 48% and among patients infected with KP-CR it was twice as high as among patients infected with KP-CS (P = 0.02). A total of 63.3% samples were classified as MDR. As for the determinants of resistance blaSHV, blaCTX-M-1, blaTEM, blaKPC, blaVIM blaNDM were the most frequent variants and none of the tested isolates presented the blaGES, blaOXA-48-like and blaIMP genes. The analysis of strains by PFGE identified a clonal group with 14 isolates and a second with only 2 isolates and 18 distinct profiles. KP-CR infection was more frequent in younger patients, associated with health care and with greater lethality. In addition, the spread of a clone between hospitals represents a challenge for the control of resistance to antimicrobials.

3
  • CLARICE RIBEIRO LIRA
  • Biperiden Effects on Ethanol Self-administration in rats

  • Advisor : RODRIGO MOLINI LEAO
  • COMMITTEE MEMBERS :
  • PAULA CRISTINA BIANCHI
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • RODRIGO MOLINI LEAO
  • Data: Jul 3, 2020


  • Show Abstract
  • Ethanol abuse is a major public health problem worldwide. The development of ethanol dependence occurs from plasticities in the central nervous system. It is known that the behavior of ethanol consumption can be modulated by cholinergic neurotransmission through nicotinic receptors. However, it is not well known about the interaction in between ethanol and muscarinic receptors. Biperiden is an anticholinergic drug that acts preferentially by blocking the muscarinic M1 receptor and has been promising for treatment of other substances of abuse. The aim of this work was to evaluate the involvement of biperiden in the motivational mechanisms and reinstatement of the ethanol seeking on the operant behavior animal model. For this, adult male Long Evans rats and self-administration boxes were used. Animals were trained to press a lever to receive an ethanol dose and for the tests the subjects were divided into the Control and Biperiden groups, and received 0.9% saline (1 ml / kg; i.p.) or different doses of biperiden (1.0; 2.5; 5.0 and 10.0 mg / kg; i.p.), respectively, 30 minutes before each test. The motivation to ethanol seeking was assessed using the Progressive Ratio (PR) schedule. Our results demonstrate a significant decrease in the breakpoint and in the number of reinforcements at doses 5.0 and 10.0 mg / kg (i.p.). In addition, the extended access protocol (“Binge”) for ethanol was carried out for a period of 5 hours. The results demonstrated a significant reduction in the search for ethanol in the animals treated with the doses 5.0 and 10 mg / kg (i.p.) when compared to the control. Later, the animals went through the extinction stage, where each pressure on the active lever was no longer reinforced by ethanol and it was performed in an different environment with tactile, visual, sound and circadian differences from the environment of the acquisition phase, which we call “Context B”. Twenty-four hours after the last extinction session, the context-induced reinstatement test was performed. It was observed that all doses of biperiden administered 30 minutes before the test were able to prevent the context-induced reinstatement of ethanol seeking.

4
  • GIULYANA EVELYN OLIVEIRA DA SILVA CAVALCANTI
  • PHENOTYPE AND GENOTYPIC CHARACTERIZATION OF ENTEROBACTERIA PRODUCERS FROM NOVA DELHI METALO-BETA-LACTAMASE (NDM) IN SALVADOR, BA

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • DAIANA DE LIMA MORALES
  • JOICE NEVES REIS PEDREIRA
  • TANIA FRAGA BARROS
  • Data: Jul 28, 2020


  • Show Abstract
  • Introduction: The spread of bacteria resistant to antimicrobials is a threat to public health. In this context, the production of the New Delhi Metalo-beta-lactamase (NDM)  enzyme is a major concern related to its ability to hydrolyze carbapenems, a specific class in the treatment of infections caused by micro-organisms resistant to multiple drugs (MDR). Since the first detection of NDM in 2009 in India, the number of patients colonized or infected with NDM-producing enterobacteria has increased worldwide. Objectives: Determine the prevalence and characterize the molecular profile of NDM-producing enterobacteria in Salvador, Bahia. Methodology: A cross-sectional study was carried out from January 2016 to December 2018 in three tertiary hospitals in Salvador. Bacterial identification and antimicrobial susceptibility tests were performed using the Vitek 2 system (BioMèrieux, France), in addition, phenotypic experimental tests were made for checking colistin resistance (drop test) and evaluation of carbapenema production (mCIM/eCIM). The presence of resistance genes was investigated by PCR, the genetic variability was verified by the total sequencing of the NDM gene. A clonal relationship between those selected was determined by PFGE and MLST. Results: In the three years of research were identified 33 cases of NDM-producing enterobacteria in a total of 301, revealing a prevalence of 11%. Cases occur more frequently in males (63.3%) and among those over 60 years of age (60%). Most isolates were obtained from blood culture (54.6%), followed by rectal swab (15.3%) and uroculture (12.2%). A more frequent species in this study was K. pneumoniae (97%), as they showed 100% resistance to ciprofloxacin, and resistance rates against beta-lactamase inhibitors (96.5%), cephalosporins (96.5%) and carbapenems (96.5%), while the lowest resistance index was against tigecycline (15.4%). The genes encoding ESBL OXA (27%), TEM (22%) and SHV (20.6%) were detected. The co-expression pattern TEM, SHV, OXA-1, CTX-M15, NDM was frequent among those indicated (27.2%). PFGE analysis showed a genetic relationship between the strains, confirmed by the MLST, which identified few ST variants. Conclusion: Here we demonstrated that NDM-producing enterobacteria are associated with an MDR profile, they exhibit rapid dissemination and the potential to cause outbreaks, representing a public health problem whose management constitutes a challenge for the infection control committee.

5
  • EMILY BATISTA PINHEIRO
  • Perfil dos anticorpos IgE e IgG4 específicos para os antígenos recombinantes Sm29 e SmKI-1 em indivíduos residentes em área endêmica em esquistossomose mansônica

  • Advisor : LUCIANA SANTOS CARDOSO
  • COMMITTEE MEMBERS :
  • LEA CRISTINA DE CARVALHO CASTELLUCCI
  • LUCIANA SANTOS CARDOSO
  • NECI MATOS SOARES
  • Data: Aug 25, 2020


  • Show Abstract
  • Schistosomiasis is a parasitic disease that affects about 200 million people and threatens more than 600 million people living in areas at risk. As a control strategy, many studies have been carried out to identify a vaccine antigen capable of promoting a partial reduction of the parasitic burden, considerably reducing the pathology and limiting the transmissibility of the disease. In this context, proteins located in the tegument of the parasite, involved in immune evasion to favor the parasite's survival, are promising candidates for the vaccine. This study aims to evaluate the production of specific IgE and IgG4 antibodies to the Sm29 and SmKI-1 antigens of Schistosoma mansoni in serum from individuals living in an endemic area in schistosomiasis. This is a follow-up study that included individuals who lived in the endemic area of schistosomiasis, municipality of Conde-BA. Serum and stool samples were collected for parasitological examinations before specific antiparasitic treatment (D0), 1 month after treatment (D30), 6 months (D180), one year after treatment (D360) and one year and six months after treatment (D540) with praziquantel. Parasitological examinations were performed using the Kato-Katz technique. The individuals were classified according to the parasitic load as negative, low load (<99 opg) and high load (> 99 opg) on D0. The levels of IgG4 and IgE antibodies specific for Sm29 and SmKI-1 antigens were assessed by indirect ELISA. Results: There was a reduction in the levels of anti-Sm29 IgG4 one year after treatment with praziquantel in the general population, in the group of negative (p <0.05), low-load and high-load individuals. Regarding anti-SmKI-1 IgG4, there was a reduction in antibody titers in the general population and in the low load group. Although no significant difference was observed in the titers of IgE anti-Sm29 and IgE anti-SmKI-1, it was observed that in D30, D180 and D360 there was a higher IgE / IgG4 anti-Sm29 ratio, when compared to the IgE / IgG4 ratio anti-SmKI-1 in the group of negative individuals, with statistical difference between the ratios at different times (D30 and D180 p <0.005; D360, p <0.05). Regarding the reinfection status (D540), no differences were observed in the specific IgE antibody titers for Sm29 and SmKI-1, in individuals who were reinfected or who remained negative in D540, but the anti-SmKI-1 IgG4 titers were higher. Among individuals who did not reinfect, we found that anti-Sm29 IgE remained higher one and a half years after treatment compared to antiSmKI-1 IgE, while there was a decrease in IgG4 titers specific for SmKI-1 compared to titers of SmKI-1. Sm29 specific IgG4. We concluded that the Sm29 antigen had a profile in negative and low-load individuals, in addition to those who did not reinfect themselves closer than expected to a protective profile for a vaccine candidate, when compared to the SmKI-1 antigen. Taking into account the protective role of IgE and the blocking action of the protective immunity of IgG4, Sm29 was characterized as a possible vaccine candidate for the control of schistosomiasis.

6
  • FELIPE PESSOA DE OLIVEIRA
  • ANTINEOPLASTIC POTENTIAL OF THE ESSENTIAL OIL FROM THE BARK OF Aniba parviflora (Meisn.) Mez (Lauraceae)

  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • DANIEL PEREIRA BEZERRA
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • GARDENIA CARMEN GADELHA MILITÃO
  • Data: Oct 30, 2020


  • Show Abstract
  • Introduction: Aniba parviflora (Meisn.) Mez is an aromatic tree belonging to the Lauraceae family and it is found in the Amazon region of several South American countries. Many pharmacological properties have been reported about the genus Aniba, however, there are few studies investigating its antineoplastic potential. Aim: To evaluate the anticancer potential of the bark essential oil of A. parviflora (EOAP). Methods: The EOAP cytotoxicity was tested in different cancer cell lines, HL-60 (human acute promyelocytic leukemia), MCF-7 (human breast carcinoma), HepG2 (human hepatocellular carcinoma), HCT116 (human colon carcinoma) and B16-F10 (mouse melanoma), and non cancer cell line, MRC-5 (human lung fibroblast) through the alamar blue assay after 72 h incubation. Next, HepG2 cells were incubated with the EOAP (5, 10 and 20 μg/mL) for 48 h, proceeding with cell cycle analysis by flow cytometry. CB-17 SCID mice were subcutaneously inoculated with HepG2 to assess the essential oil (40 and 80 mg/kg) in vivo antitumor potential. Results: Chemical analysis identified linalool (16.3 ± 3.15%), α-humulene (14.5 ± 2.41%) and δ-cadinene (10.2 ± 1.09%) as the major components present in the EOAP. The IC50 values displayed for EOAP ranged from 9.05 to > 50 μg/mL for HepG2 and HCT116 cells, respectively. The IC50 value found for the non-cancer cell MRC-5 was 30.46 μg/mL. The EOAP at a concentration of 20 μg/mL induced DNA fragmentation in HepG2 cells after 48 h incubation, suggesting cell death. In the in vivo xenograft study, it was demonstrated that mice presented reduced tumor weight after treatment with the highest dose of EOAP (80 mg/kg). Conclusion: The essential oil from the bark of A. parviflora might be a valuable source of new substances for cancer treatment, taking into account its cytotoxic and antitumor action.

7
  • OTÁVIO AUGUSTO CARVALHO DE OLIVEIRA SANTOS
  •  Type B natriuretic peptide (BNP, pro-BNP, and NT-pro-BNP) as 
    a biomarker of heart failure to be incorporated into the clinical practice of the Unified Health 
    System – SUS/Brazil: evaluation of health technologies.
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • RICARDO DAVID COUTO
  • ANA LEONOR PARDO CAMPOS GODOY
  • PABLO DE MOURA SANTOS
  • Data: Nov 26, 2020


  • Show Abstract
  • Introduction: Heart failure is the final clinical manifestation of several cardiac pathologies. BNP is a neurohormone secreted by the ventricles in response to volume expansion and cardiac overload and was considered an important biochemical marker for diagnosis, evaluation, and prognosis of patients with congestive heart failure. Objective: To show the feasibility of implementing the determination of type B natriuretic peptide (BNP, pro-BNP and NT-pro-BNP) in the clinical practice of the Unified Health System – SUS/Brazil, given the association of these biomarkers with the clinic of patients with heart failure and the economic viability added by this diagnostic support procedure. Methodology: Observational, cross-sectional study, carried out with a convenience sampling, from the collection of data in medical records and in the laboratory information system from patients with congestive heart failure, admitted to the Hospital da Bahia, as well as data from patients from the Ana Nery Hospital / UFBA, and Clínica Santa Helena/Camaçari, Bahia. Results: From 55 patients evaluated, 89% (n = 49) had high BNP values, 71.4% (n = 35) with heart failure. The remainder (28.6%), despite not having heart failure, BNP was above the cut-off values, due to heart disease, in addition to other associated comorbidities that also elevate BNP, such as sepsis, kidney and respiratory failure. Regarding implementation costs, quotations in the national market, the values for determining BNP, (single test) varied between 48.50 and 177.00 reais. Depending on the methodology and number of tests to be realized, if the BNP kit can be adequately purchased, to access the estimated values, excellent cost-benefit ratio would be achieved, that is, between 13.64 and 27.00 reais each test determination. Conclusion: Therefore, based on all currently available information, implementing the BNP/NT-proBNP biomarkers determination within the scope of SUS is economically viable, will speed up diagnosis, prognosis, better patient follow-up and clinical conduct so far implemented for heart failure.

8
  • GABRIEL SILVA LIMA
  • DIGITAL PHARMACOLOGY: DEVELOPMENT OF A APPLICATION AS AN EDUCATIONAL TOOL FOR THE PHARMACOLOGY FIELD
  • Advisor : DENIS DE MELO SOARES
  • COMMITTEE MEMBERS :
  • DENIS DE MELO SOARES
  • MARCELO SANTOS CASTILHO
  • THAIS RODRIGUES PENAFORTE
  • Data: Dec 16, 2020


  • Show Abstract
  • Mobile technology for students' lives has increased in importance. Devices such as smartphones, tablets and e-book readers connect users to the world immediately, increasing information accessibility and enabling interaction between them. Assessing this reality, mobile technologies to teach and learn has grown in education. In this context, many universities worldwide have started using smartphones to promote learning and disseminate information. To support technological resources for transmission of content, psychology through theories of learning investigates since last century possible contributions brought by machines into learning process. Based on gamified systems and Skinner's radical behaviorism, using technological resources available, this work aimed to develop, on Android platform, an educational application to access contents in the pharmacology field. Two versions were developed and published on Google Play. The necessary steps to develop the first version, with exception of programming, were performed by project creator. To launch the second version, or update, all steps were performed by creator. It was necessary to learn highlevel Java programming language. The initial version features a database with 205 questions and 969 images. In view of suggestions made by the board of examiners and users on app's comments page on Google Play, modifications were made. The current version includes a database with 310 questions and 810 images, as well as classes, initially in PDF format. This application can be used, above all, as a complement to classroom. Activities can be used for practice drills and to access pharmacology materials. Finally, this work aims to follow technological advances that can contribute to other educational innovations.

9
  • FLORISVALDO DA SILVA RAMOS
  • PHYTOCHEMICAL STUDY AND EVALUATION OF CYTOXOTIC POTENTIAL OF Paubrasilia echinata  LAMK., FRONTOF GLIAL TUMOR CELL LINES

  • Advisor : EUDES DA SILVA VELOZO
  • COMMITTEE MEMBERS :
  • DANIELA SALES ALVIANO MORENO
  • EUDES DA SILVA VELOZO
  • LUZIMAR GONZAGA FERNANDEZ
  • Data: Dec 22, 2020


  • Show Abstract
  • Glial and stem cells are among the main ones in the human brain. The latter can undergo changes and multiplication, giving rise to glioblastoma multiform (GBM). The Paubrasilia genus has metabolites derived from polyphenols, steroids, diterpenes and triterpenes that exhibit analgesic, antibacterial, anti-inflammatory, antioxidant, antiproliferative, and immunomodulatory activities. This work aimed to isolate, purify and identify the components present in the bark of Paubrasilia echinata Lamk. in a bioguided way to obtain substances with antitumor activity against glioma cells in vitro. For this purpose, the cytotoxic potential of isolated fractions and compounds (F001) and (E002) was identified in different extracts of Paubrasilia echinata to decreasing the activity viability or migration of tumor from rat (C6) and human (U251) glioma cells in vitro. C6 and U251 cells were used in Dulbecco's Modified Eagle's Medium (DMEM) and in medium with dimethylsulfoxide (DMSO) as controls. The methodologies applied were 3- [4,5-dimethylthiazol-2-yl] -2,5-dimethyltetrazolium Bromide test (MTT), EC50 determination of the concentration that kills 50 % of cells (EC50) and Cell Migration Inhibition (CMI) test. The substance F001 had no significant activity in C6 and U251 cells during cell viability tests with MTT at concentrations between 1.2 - 200 µM. However, the substance F001 inhibited cell migration in concentrations at 40, 60, 120 and 200 µM in the BMI test. The substance E002 presented a significant difference in relation to the control for the cell viability test with MTT at concentrations above 19 µM (EC50 = 38 µM). The CMI test demonstrated that there was no cell growth for the same concentrations. The results allow us to conclude that the F001 substances extracted from P. echinata inhibited cell migration, while E002 exhibited cytotoxicity to C6 and U251 rat and human glioma cells, respectively.

2019
Dissertations
1
  • MARCELO TELES BASTOS RIBEIRO
  • MOLECULAR EPIDEMIOLOGY OF STAPHYLOCOCCUS AUREUS INFECTIONS IN SALVADOR, BA

  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • JOICE NEVES REIS PEDREIRA
  • DENIS DE MELO SOARES
  • LUCAS MIRANDA MARQUES
  • Data: Jan 21, 2019


  • Show Abstract
  • INTRODUCTION: Methicillin-resistant Staphylococcus aureus (MRSA) infections are a public health problem because of the adaptive nature and ability to develop resistance to antibiotics. People infected with HIV are at higher risk of acquiring this type of drug, which requires careful and specific clinical treatment during the course of the disease. OBJECTIVE: The objective of this study is to describe S. aureus infections and to characterize the isolates by phenotypic and genotypic methods. MATERIAL AND METHODS: An observational, prospective study was conducted between June/2016 and March/2018. Clinical-epidemiological data were collected through medical charts review. The microorganisms were identified and submitted to the antimicrobial susceptibility test using the VITEKII automated system (Biomeuriex, France) and by the Kirby-Bauer method; the resistance gene (SCCmec) and virulence (PVL) were investigated by PCR and the comparison of the pulsed field (PFGE) electrophoresis profile through a dendogram generated by computerized analysis (GelCompar II software). RESULTS: In the study period, 40 patients developed S. aureus infection, with 19 (47.5%) bloodstream infection, 4 (10%) catheterrelated infection, 6 (15%) respiratory infection, 1 (2.5%) urinary infection and 10 (25%) tissue and soft tissue infections. In the sample, males were predominant (31/40, 77.5%) and median age 36 years. A total of 18 (45.0%) of the cases occurred in patients infected with HIV. The frequency of methicillin-resistant isolates (MRSA) by phenotypic methods was 62.5% (25/40), 40% (16/40) to clindamycin, 60% (24/40) erythromycin, and five cases were test D positive (12.5%). Of the total cases with MRSA profile, 18/25 (72%) were confirmed through PCR, and the following SSCmec-cassettes were identified: type I in 8 (20%), type III in 8 (20%), type IV in one (2.5%) and type V in one (2.5). The MRSA profile was not associated with patients infected with HIV, (p = 0.49, OR: 0.64, 95% CI: 0.18-2.25). Patients who progressed to death were admitted to the intensive care unit (p = 0.004, OR: 10.56 CI 95%: 1.90-58.53), had HIV infection (p = 0.002, OR: 12.50 CI 95% CI: 1.17-21.39), isolate with erm A genotype (p = 0.01, 95% CI: 2.23-70.19), resistance to clindamycin (p = OR: 6.44 95% CI: 1.44-28.89) and SCCmec type III genotype (p = 0.04, OR: 5.95 95% CI: 1.13-31.27). The gene encoding the PVL toxin was identified in three cases (7.5%) with different clonal profile and SCCmec genotypes. The analysis of the electrophoresis profile generated by the PFGE technique revealed five clonal groups (A, B, C, E, O). CONCLUSION: Local knowledge of the epidemiological and molecular aspects of staphylococcal infections provides baselines to further improve the prevention, treatment and control of S. aureus infections, especially among vulnerable patients.

2
  • FÁBIO MOTA SILVA
  • REAL TIME PCR WITH "MOLECULAR BEACONS" HYBRIDIZATION PROBE FOR CANDIDA SPP DETECTION IN HEMOCULTURE
  • Advisor : TANIA FRAGA BARROS
  • COMMITTEE MEMBERS :
  • TANIA FRAGA BARROS
  • JOICE NEVES REIS PEDREIRA
  • BARBARA MARIA PARANA DA SILVA SOUZA
  • Data: Feb 15, 2019


  • Show Abstract
  • Candidemia, a bloodstream infection caused by Candida yeasts, has become a growing problem in tertiary care hospitals, in which patients suffering from the disease have high morbidity and mortality rates due to their difficulty in the early diagnosis of the disease. Thus, aiming at new methods for the detection of Candidemia, the objective of our study was to standardize real-time PCR technique with Molecular Beacons hybridization probe for the detection of Candida spp. in blood culture. To this end, a target sequence of the rDNA ITS region (ITS1-5.8S-ITS2) was chosen and from there a "Molecular Beacons" hybridization probe was developed for conducting the real-time PCR assays. The probe was standardized with DNAs from standard strains and tested with DNAs extracted from blood cultures from patients of a public hospital and a private hospital located in Salvador, Bahia. The "Molecular Beacons" hybridization probe detected the presence of yeasts of the Candida genus, including the C. albicans, C. glabrata, C. parapsilosis, C. tropicalis species in positive blood cultures of patients with candidemia, as well as yeasts of the genus Saccharomyces. The probe did not detect the presence of yeasts of the genus Cryptococcus and of the Kodamaea genus in positive blood culture of a patient with these yeasts, did not detect the presence of bacteria in positive blood cultures of patients with bacteremia, nor did it to promote positive reaction in blood cultures of patients without invasive infection. Compared with the hydrolysis probe developed by Amaral (2017), the hybridization probe "Molecular Beacons" developed with the same nucleotide sequence may demonstrate greater specificity. Therefore, the "Molecular Beacons" hybridization probe developed can be used in the real-time PCR assay for rapid detection of Candida spp. directly from blood cultures in the diagnosis of candidemia.

3
  • RAFAELA GOMES ALVES COSTA
  • POTENCIAL ANTINEOPLÁSICO DO ÓLEO ESSENCIAL DAS FOLHAS DE Guatteria megalophylla DIELS (ANNONACEAE)

  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • CRISTIANE FLORA VILLARREAL
  • DANIEL PEREIRA BEZERRA
  • JUNIA RAQUEL DUTRA FERREIRA
  • Data: Feb 22, 2019


  • Show Abstract
  •  

    Introduction: Essential oils obtained from several plant species have been considered as potential sources of biologically active substances, presenting several therapeutic properties. The Guatteria megalophylla Diels (Annonaceae) plant is a tree native to the Brazilian and Peruvian Amazon rainforest. Its cytotoxic potential had not yet been described in literature; however, within its family there have already been reports of several species with pharmacological potential, including antitumoral activity. Objective: To evaluate the antineoplastic potential of the essential oil from G. megalophylla leaves. Methods: The chemical composition of the essential oil was determined by gas chromatography with flame ionization and mass detectors. The cytotoxic activity of the essential oil was tested in different human cancer cell lines, HL-60 (acute promyelocytic leukemia), MCF-7 (breast carcinoma), CAL27 (tongue squamous cell carcinoma), HSC-3 (tongue carcinoma), HepG2 (hepatocellular carcinoma) and HCT116 (colon carcinoma), and non-cancer cell line, MRC-5 (human lung fibroblast) by the alamar blue method after 72h incubation. Subsequently, HL60 cells were incubated for 24 and 48h with the essential oil (10, 20 and 40 μg/mL) and the number of viable cells was determined by the trypan blue exclusion assay, and cell cycle analysis, mitochondrial transmembrane potential, as well as quantification of reactive oxygen species (ROS) were evaluated by flow cytometry. Results: The essential oil was the major constituent of spathulenol and showed IC50 values for cancer cells ranging from 12.5 to 39.5 μg/mL for HL-60 and CAL27, respectively. The IC50 value for the MRC-5 non-cancer cells was 29.8 μg/mL. The cytotoxic effect of the essential oil was confirmed by the tripan blue exclusion assay, where it showed a reduction in the number of viable cells. It was also observed that the essential oil induces internucleosomal DNA fragmentation after 24 and 48h and induces depolarization of the mitochondrial transmembrane potential, but did not induce ROS production. In addition, the essential oil inhibited the in vivo development of HL-60 cells inoculated in C.B-17 SCID mice. Tumor mass inhibition rates were 16.63% at 50 mg/kg concentration and 48.79% at 100 mg/kg concentration. Conclusion: The essential oil of G. megalophylla leaves may be a promising phytotherapeutic substance for the treatment of several types of cancers since it exhibits cytotoxicity for different carcinogenic cell lines, in addition to presenting antitumor activity in vivo.

4
  • DOURIVALDO SILVA SANTOS
  • Pharmacological Characterization of Antinociceptive Properties of Spirulina platensis LEB 18
  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CRISTIANE FLORA VILLARREAL
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • JANICE IZABEL DRUZIAN
  • Data: Mar 22, 2019


  • Show Abstract
  • Pain has a protective physiological function; however the intense or persistent pain causes suffering to the patient, requiring medication intervention. First-line drugs for pain control, opioids and nonsteroidal anti-inflammatory drugs can cause serious side effects, especially for prolonged use. Thus, adequate pain control still depends on the development of new drugs, more effective and with fewer side effects. Natural products are a promising source of bioactive substances. Spirulina platensis (SP) is a cyanobacterium that produces a wide variety of active metabolites, and has aroused the interest of the pharmaceutical and food industry. In this work, the antinociceptive properties SP were established in experimental models, investigating their possible mechanisms of action. Initially, the antinociceptive effect of SP was investigated in an inflammatory pain model. Oral administration of SP (50-400 mg / kg) produced dose-dependent inhibition of inflammatory allodynia (p <0.01) and paw edema (p <0.001) in the Freund's Complete Adjuvant (CFA) model in mice. Treatment with SP (200 mg / kg) modulated the balance of pro- and anti-inflammatory cytokines, i.e., reduced levels of hyperalgesic cytokines (TNF-α, p <0.05, IL-1β, p<0.01) and elevated those of IL-10, an anti-inflammatory cytokine (p <0.05). Treatment with SP (200 mg / kg) in IL-10 knockout mice (IL-10 / KO) did not prevent inflammatory allodynia nor the formation of paw edema induced by CFA. This result indicates that the SP-induced anti-inflammatory effects are mediated by IL-10 production. Next, we investigate whether the SP-induced antinociception present a centrally mediated component. Treatment with spirulina (100-400 mg / kg) dose-dependently increased the latency time in the tail-flick test. The antinociceptive effect of SP in the tail flick test was partially prevented by pre-treatment with naloxone (5 mg / kg), a non-selective opioid receptor antagonist (p <0.05). Thus, it is possible to propose that SP-induced antinociception has a central mediating component involving activation of opioid pathways. Mice treated with spirulina (400 mg / kg) showed no motor deficit in the rota-rod test, corroborating data from the nociceptive tests. In conclusion the present study showed that SP has relevant antinociceptive properties. Importantly, the anti-inflammatory and antinociceptive actions of this cyanobacterium appear to be independent, possibly mediated by IL-10 and opioid pathways, respectively. Considering that Spirulina platensis is already approved for human use, this study validates its clinical indication as an anti-inflammatory and allows us to propose a new as an analgesic.

5
  • ISIS GOMES DOS SANTOS
  • CHEMICAL-PHARMACEUTICAL ANALYSIS AND STABILITY STUDIES OF HYDROHYUREA IN RAW MATERIAL AND CAPSULES

  • Advisor : EDITH CRISTINA LAIGNIER CAZEDEY
  • COMMITTEE MEMBERS :
  • EDITH CRISTINA LAIGNIER CAZEDEY
  • JOSE ANTONIO MENEZES FILHO
  • ANIBAL DE FREITAS SANTOS JUNIOR
  • Data: Apr 30, 2019


  • Show Abstract
  • Hydroxyurea (HU) is the therapeutic class of antimetabolites and acts on the cell cycle by inhibiting the ribonucleotide reductase enzyme present in DNA biosynthesis and maintaining it in the S phase. This drug is used in the treatment of neoplastic diseases such as myeloid leukemia, psoriasis and has recently shown great clinical efficacy in the treatment of sickle cell disease, being the only drug capable of reducing vascular occlusion effects and improving the quality of patients’ life. Despite the clinical importance of HU, the Brazilian Pharmacopoeia 5th edition (2010) does not present a monograph intended for this drug, nor does it exist in the scientific literature and official compendiums stability indicating analytical methods (SIAM) for HU. The development and validation of analytical methods for this molecule is justified by its therapeutic potential and the need for methods that prove its quality, with agility, low cost and lower environmental impact. In this work the qualitative analysis of the raw material and finished product was carried out, verifying the organoleptic characteristics, uniformity of weight for the capsules, identification by infrared (IR) and ultraviolet (UV) spectrophotometry and by high performance liquid chromatography  with photodiode array (HPLC - PAD). In addition, two analytical methods were developed and validated for the quantification of HU raw material and in capsules using UV spectrophotometry and HPLC-PDA, a technique used also in the forced degradation studies. The UV spectrophotometric method demonstrated that the most suitable solvent for the quantitative analysis of HU was water and the most appropriate wavelength was 213 nm. The method was selective, precise, accurate and robust. For the SIAM, chromatographic separation was obtained with Phenomenex® C18 column (250 x 4.6 mm, 5μm); the mobile phase was composed by water and ethanol (95:05, v/v), in isocratic mode. The flow rate was 0.5 mL/min and the wavelength was 214 nm. Forced degradation studies evidenced that HU degraded in oxidative medium, in basic and acid hydrolysis and was stable in thermal degradation. The SIAM for the quantification of HU was selective, linear, precise, accurate, sensitive and robust. The results of the methods development by UV spectrophotometry and HPLC-PAD for the quantitative analysis of HU were positive and met national and international quality requirements.

6
  • ALEX BRUNO DA SILVA SOUZA
  • Evaluation of cytokines in alcoholic patients Strongyloides stercoralis infected

  • Advisor : NECI MATOS SOARES
  • COMMITTEE MEMBERS :
  • FERNANDA WASHINGTON DE MENDONCA LIMA
  • JOELMA NASCIMENTO DE SOUZA
  • NECI MATOS SOARES
  • Data: May 16, 2019


  • Show Abstract
  • Strongyloides stercoralis infection prevalence is high in alcoholic patients. Excessive alcohol consumption increases the levels of endogenous corticosteroids, which mimic the effect of the ecdysone, a parasitic hormone which stimulates the differentiation of rabditoid into infectious filarioid larvae. The immune response to strongyloidiasis in immunocompromised patients is usually associated with a Th2 cellular response profile. The aim of this study was to evaluate serum levels of cytokines (INF-γ, IL-2, IL-4, IL-5, IL-10, IL-15, IL-17) in alcoholic patients infected with S. stercoralis and to correlate its levels with the parasite load. S. stercoralis frequency was evaluated in 336 alcoholic patients attended at the Centro de Acolhimento e Tratamento de Alcoolistas (CATA), Obras Sociais Irmã Dulce, Salvador, BA, from April 2017 to August 2018. The parasitological diagnosis was performed by three methods: agar plate culture, Baermann-Moraes and spontaneous sedimentation in, at least, two fecal samples from each patient on alternate days. Serum cytokines levels were determined in 88 sera from S. stercoralis  infected (n = 24) and non-infected (n = 24) alcoholic patients and S. stercoralis  infected (n = 20) and non-infected (n = 20) non-alcoholic individuals (n = 20). S. stercoralis infection frequency in alcoholic patients was 16.1% (54/336). Among the evaluated cytokines, a statistically significant elevation in IL-10 and IL-15 levels was observed in patients compared to S. stercoralis infected non-alcoholics individuals (p <0.05). A negative correlation was demonstrated between the concentration of INF-γ and IL-17 with the parasite load of alcoholic patients. Thus, it is probably that alcohol is inducing the simultaneous elevation of both pro-inflammatory and regulatory cytokines, IL-15 and IL-10, respectively. The negative correlation of INF-γ and IL-17 concentrations with the parasite load probably reflects on the modulation of the T reg cells in the control of the strongyloidiasis.

7
  • SARA EMI MATOS MENDES FERREIRA
  • EVALUATION OF THE INVOLVEMENT OF THE FRAGILE PROTEIN OF MENTAL RETARDATION X (FMRP) IN THE DEVELOPMENT OF ETHANOL DEPENDENCE: FROM VIRTUAL SCREENING TO BEHAVIOR.

  • Advisor : RODRIGO MOLINI LEAO
  • COMMITTEE MEMBERS :
  • RODRIGO MOLINI LEAO
  • DENIS DE MELO SOARES
  • REJANE CONCEICAO SANTANA
  • Data: May 24, 2019


  • Show Abstract
  • Drug abuse is currently a global public health problem. The abusive consumption of ethanol promotes neuroplasticity and changes in balance between excitatory and inhibitory neurotransmitters, as well as altering proteins involved in gene expression, altering neuronal excitability. Chronic exposure to ethanol is capable of generating tolerance and dependence, although some aspects of dependence may occur relatively rapidly in response to acute administration, most of the changes occurring gradually occur over time in response to prolonged exposure to the drug. Chronic ethanol treatment has been shown to enhance the phosphorylation of Fragile X Mental Retardation Protein (FMRP) protein. From the increase of its activity, it was observed an increase in gene transcription and protein plasticity in glutamatergic synapses in regions important for the development of ethanol dependence. The objectives of this study are: 1. To develop a new molecule with FMRP protein antagonist activity and its modeling, using bioinformatics tools 2. To express the protein by cloning in vitro and verify the optimal conditions for its expression; 3. Evaluation of neuroadaptations through behavioral analysis of the development and expression of locomotor sensitization in the open field with male Swiss mice (30g) from the administration of ethanol (EtOH 2.2g / kg, ip) and saline (SAL 0.1ml / kg; ip) (Ethics committee Protocol 110/2016); 4. Evaluation of neuronal activation by immunohistochemistry of the regions of the Prefrontal Cortex, Nucleus Acúmbens and Amídala. Our results show the expression of FMRP in the conditions of high concentration of IPTG (1M) and high temperature in short period (37 ° C / 4hrs); In the behavioral analysis it was observed that animals treated with EtOH (chronic) showed a progressive increase in locomotor activity (initial mean of 25.41m and final mean 62.17m) during the development of locomotor sensitization and also showed greater locomotor activity than the control groups, which maintains a constant locomotion pattern, between 38 and 33 meters (average of day 1 and day 21 respectively). The chronic group also had greater locomotor activity in the expression of sensitization when compared to the other control and acute groups (mean 42.02). It was also possible to identify a tendency in the activation of the brain regions NAc and CPFm in the brains of the animals after treatment with the drug.

8
  • LUCIANA SANTA BARBARA BITTENCOURT

  • FUNGEMIA IN A HIGH SCHOOL UNIVERSITY HOSPITAL: EPIDEMIOLOGICAL AND DIAGNOSTIC SURVEY OF REAL-TIME PCR CANDIDEMIA.

  • Advisor : TANIA FRAGA BARROS
  • COMMITTEE MEMBERS :
  • ANA PAULA DE OLIVEIRA MENEZES
  • DENIS DE MELO SOARES
  • TANIA FRAGA BARROS
  • Data: Aug 23, 2019


  • Show Abstract
  • Invasive fungal infections have been increasingly common in recent decades, becoming one of the leading causes of morbidity and mortality in immunocompromised patients. Geographic differences in species distribution and in vitro susceptibility patterns to antifungal agents reaffirm the importance of conducting epidemiological surveillance studies of fungemia. The treatment of invasive fungal infections has been hampered by the inability to diagnose the infection at an early stage of the disease. With the need for faster strategies in the process of detection and identification of these pathogens, real-time polymerase chain reaction technology has improved the viability of the results. Therefore, the objective of this study was to make an epidemiological survey of patients with fungemia at Professor Edgard Santos University Hospital, Salvador, Bahia, from January 2018 to January 2019, diagnosing candidemia through real-time PCR assays. A total of 6204 blood cultures were processed with 9.5% positivity; of these, 91.5% positive for bacteria and 8.5% positive for fungi, being the yeast in fourth place among the most prevalent isolates. 24 individuals presented 25 isolates in fungi positive blood culture: 72% from Candida yeast, 20% Cryptococcus, 4% Histoplasma capsulatum and 4% without identification. Most patients were 58.3% male (14/24), with a mean age of 39.8 years and a median of 56.5 years. It was found that the highest prevalence was in patients with HIV positive and gastrointestinal disease, both with a percentage of 33.3%, followed by renal failure with 29.2%, neoplasia with 25% and systemic arterial hypertension with 20.8. %. Real-time PCR assays show that the molecular beacons hybridization probe of the intergenic region was fungus specific, detecting positive blood cultures for C. glabrata, C. albicans, C. krusei, C. tropicalis and C. parapsilosis, but further studies are still needed to increase the specificity of the probe, eliminating the detection of genera Aspergillus and Saccharomyces. Candida detection by real-time PCR assay was possible in blood culture samples and not in blood samples.

9
  • RÔMULA BETÂNIA MENDES ALECRIM DA ROCHA
  • EXPOSURE TO SMOKE FROMM WOOD BURNING AND LEAD OF THE POTTERY WORSHOPS AND HEALTH EFFECTS ONSCHOOL-AGED CHILDREN IN MARAGOGIPINHO, BAHIA
  • Advisor : JOSE ANTONIO MENEZES FILHO
  • COMMITTEE MEMBERS :
  • JOSE ANTONIO MENEZES FILHO
  • ANA LEONOR PARDO CAMPOS GODOY
  • MANSUETO GOMES NETO
  • Data: Aug 28, 2019


  • Show Abstract
  • ntroduction: Air pollutants are crucial for the disease process, especially with regard to respiratory diseases. Noteworthy is the burning of biomass that generates numerous compounds, including particulate matter (PM), one of the main pollutants that has environmental importance and toxicological interest. Objective: To evaluate the exposure to smoke from wood burning and lead (Pb) from the potteries of Maragogipinho, Aratuípe, Bahia and to investigate its association with health effects on school-aged children from this locality. Methodology: This cross-sectional study included 72 children aged 6 to 14 years, of both sexes, enrolled in elementary schools in Vila de Maragogipinho and classified as moderate exposure (MEx) and low exposure (BEx) according to the deposition rates of Pb in the dust. These were compared with 71 children from schools located in the urban center of the municipality Aratuípe, called control group (CG). Sociodemographic data, anthropometric parameters were recorded and the frequencies of asthma, rhinitis and allergies were determined using the ISAAC questionnaire. Data on blood lead levels (PbS) were collected from the CAS (Lead, Environment and Health) study. One hundred and eighteen children (82.5%) had their blood pressure (BP) measured and underwent respiratory function test using a portable digital spirometer. Results: Of the total children evaluated (n = 143), 77 (53.85%) were female, the median age in group BEx was 8.41 (6.7 - 13.87) years, group MEx was 10.87 (5.9 - 14.04) and in the GC group it was 11.91 (7.65 - 14.17) years. The blood lead had a median of 2.3 μg / dL (ranging from 0.1 μg / dL to 21.3 μg / dL). The children's BP presented values within the normal range, with no significant difference for systolic and diastolic BP in both groups (p = 0.088 and p = 0.077, respectively). However, in the MEx group, a slight increase in the 95th and 99th percentiles of both systolic and diastolic BP was detected in 4.9% of children. However, in the CG, a slight increase was observed in the 90th, 95th and 99th percentiles for systolic BP with 5.9% of children and a slight increase only in the 99th percentile for diastolic BP with 1.5% of children, which can be classified as hypertensive (HP). There was no significant correlation between the systolic and diastolic BP variables and PbS levels. A high frequency of asthma symptoms was observed in 65% of children in the MEx group, 44.1% in the BEx group and 50% in the CG, but there was no significant difference (p = 0.163) between the groups. The frequency of allergic rhinitis was 38.2% in the BEx group, 47.5% in the MEx group and 30.9% in the CG. The results for respiratory function parameters such as forced vital capacity (% FVC) and forced expiratory volume in one second (% FEV1) tended to decrease with increased of blood lead in the BEx and MEx groups, but with low values (R2 = 0.006 and R2 = 0.061; R2 = 0.013 and R2 = 0.056 respectively). Blood lead levels were significantly correlated (p <0.05) with Z-score Height / Age, Weight and BMI and with respiratory parameters %FEF25-75 and %PEF.  Conclusions: These findings confirm growing evidence that air pollution may be directly linked to health problems for the population, especially the child population. For a better stratification of the results it would be necessary to carry out a clinical monitoring of the respiratory function measurement of these children and to quantify the PM concentrations in several points of this locality.

10
  • VICTOR OTERO MARTINEZ
  • Toxoplasma gondii INFECTION IN SCHOOLS FROM A COMMUNITY OF KILOMBALL TRADITIONS: SOCIAL AND ENVIRONMENTAL DETERMINANTS AND EFFECTS ON BEHAVIOR
  • Advisor : JOSE ANTONIO MENEZES FILHO
  • COMMITTEE MEMBERS :
  • JOSE ANTONIO MENEZES FILHO
  • LUCIANA SANTOS CARDOSO
  • CHRISSIE FERREIRA DE CARVALHO
  • Data: Aug 30, 2019


  • Show Abstract
  • Toxoplasma gondii (T. gondii) infection is highly prevalent in the Brazilian population. Although considered a self-limiting infection in immunologically healthy individuals, recent studies suggest that this pathogen may modify human behavior. The objective of this study was to evaluate the seroprevalence of T. gondii infection, its social and environmental determinants and the possible effects on the behavior of school children in Aratuípe, Bahia. Information was collected from 142 children (6 to 14 years old) of both sexes, enrolled in the municipal schools of Aratuípe, Bahia, Brazil. Laboratory determinations were performed by Enzyme-linked Immunosbsorbent Assay (ELISA). Risk factors were analyzed using a semi-structured questionnaire and the assessment of child behavior was made from the Child Behavior Check List (CBCL) applied to parents or guardians. As it is a region of recognized production of ceramic utensils, intense use of lead oxide has already been evidenced in the glazing process, so data on blood lead levels (BLL) in children were used as a covariate, since this heavy metal is a well-known neurotoxic agent, associated with attention deficit hyperactivity disorder. Multivariate models applying logistic regression were used to define the determinants significantly associated with chronic T. gondii infection, as well as to define its association with behavioral outcomes in children. Seroprevalence for anti-T. gondii IgG antibodies was 43.7% (95% CI, 35.8% - 51.9%), children positive for T. gondii specific antibodies had a 2.34 times greater chance (95% CI, 1.13 - 5.25) of having been hospitalized due to infections, children with chronic T. gondii infection were less likely to report contact with soil, the frequency of the other risk factors surveyed showed no significant differences between seropositive and seronegative individuals. Regarding child’s behavior, 21.8% of the children had problems with clinical classification. There was an association between chronic T. gondii infection and total behavioral problems (OR = 2.50; 95% CI 1.06 - 5.88), internalizing spectrum problems (OR = 4.35; 95% CI 1.11 - 17.14) and disobedience to rules (OR = 2.61; 95% CI 1.12 - 6.05). Although no association was observed between high levels of BLL and behavioral effects, a possible interaction between toxoplasmosis prevalence and lead exposure was detected. Children with above median BLL and IgG anti-T. gondii positive showed a 5.51-fold increase (OR = 3.47, 95% CI 1.75 - 17.38) in the chance of disobedient behavior. The results suggest that in the municipality children there is a high prevalence of chronic toxoplasmosis, and that the parasite infection may be contributing to the high rates of behavioral changes. In addition, these findings are the first evidence of an association between chronic T. gondii infection and elevated blood lead levels with neurobehavioral effects in children.

     

11
  • MATHEUS DE JESUS BANDEIRA
  • Co-exposure to lead and smoke from wood burning and association with genotoxic effects in potters from Maragogipinho, Bahia

     

  • Advisor : JOSE ANTONIO MENEZES FILHO
  • COMMITTEE MEMBERS :
  • JOSE ANTONIO MENEZES FILHO
  • TANIA MASCARENHAS TAVARES
  • MARCO ANTONIO VASCONCELOS REGO
  • Data: Sep 12, 2019


  • Show Abstract
  • Maragogipinho is a traditional pottery center located in the Reconcave Region of Bahia, in which the use lead (Pb) as a flux in the glazing process of pottery utensils brings risks to the artisans, the community in general and the environment. Pb is a toxic metal already known in occupational and environmental settings, affecting several organs and systems, especially the nervous and circulatory, besides been classified as a probable human carcinogen. Thus, this work aims to evaluate the determinants related to occupational exposure to Pb and wood smoke, and the association with genotoxic effects in Maragogipinho artisans. This is a cross-sectional study with workers of both sexes, from 16 years of age and older (N = 85), compared with workers from various occupations residing in the urban area of Aratuípe town (N = 50). To assess Pb exposure, Pb deposition rate in dust (PbP) at the pottery workshops was determined, venous blood collected for PbS determination and urine for d-aminolevulinic acid (ALA) measurement, as an effect biomarker, and to estimate the exposure to smoke of wood burning, 1-hydroxypyrene (1-OHPy) was measured. The genotoxic effect was evaluated by frequency of micronucleus (MN) and other chromatin abnormalities in oral mucosa cells. Occupational exposure and socioeconomic data were collected with semi-structured questionnaires applied by interview. The study was approved by the UFBA Research Ethics Committee. The medians levels of PbS and PbP (min - max) found for the exposed group were 7.9 µg/dL (0.9 - 49.8) and 1,316 μg/m²/30 days (7 - 1,562,626), respectively. For the unexposed group PbS and PbP levels were 1.5 μg/dL (0.1 - 19.8) and 64 μg/m²/30 days (52 - 200), respectively. PbS levels were positively and significantly correlated with years of occupation (Rho = 0.351; p <0.001), Pb oxide use (Rho = 0.191; p = 0.027), male gender (Rho = 0.434; p <0.001), PbP (Rho = 0.656; p <0.001) and inversely correlated with Schooling (Rho = -0.333; p <0.001). The mean ALA-U concentrations were 3.30 ± 1.77 mg/g creat. for the exposed group and 3.28 ± 1.59 mg/g creat. for the unexposed group, these levels were below the ones reported in literature for workers exposed to Pb. There was no significant correlation between ALA-U and PbB levels. The following determinants of elevated blood lead (PbS > 10 μg / dL) based on binary logistic regression were identified: Occupation (OR = 168.995; 95% CI = 3.954 – 7223.082), Pb use in workplace (OR = 14.536; 95% CI = 2.435 – 86.780). On the other hand, having completed high school was identified as a protective factor (OR = 0.041; 95% CI = 0.004 – 0.437). The geometric means of 1-OHPy concentrations for the exposed and unexposed groups were 1.13 (0.04 - 6.41) µg/g creatinine and 0.47 (0.04 - 5.97) µg/g creatinine, respectively. Regarding genotoxic effects, it was observed a significant correlation between PbS levels and the frequency of nuclear pycnosis (Rho = 0.363; p = 0.006), while 1-OHPy was linearly associated with Broken Egg frequency (β = 0.598 [95% CI -0.032 - 1.227]), adjusted for Age and Smoking. Considering only the exposed group, LogPbS had a borderline correlation with the frequency of pycnosis (β = 2.495 [95% CI -0.216 - 5.206]), when adjusted for smoking, age and years of occupation. The frequencies of MN found were much lower than those reported in literature. In conclusion, it was possible to verify that there is an excessive occupational exposure among pottery artisans in Maragogipinho district, both to Pb and to the firewood smoke. Elevated dust deposition rates of Pb, high blood lead levels and smoke exposure may be associated with increased frequency of genotoxic endpoints. The values observed in this study exceed the levels considered occupationally safe and those reported in the literature, alerting to the need to implement occupational hygiene measures and improvements in the working conditions of these artisans, especially the replacement of lead oxide in the pottery glazing process.

12
  • ROSEMILIA RIBEIRO DA CUNHA
  • LIFTING PLAN FOR PURPOSES MEDICINALS COLLECTED IN BAHIA AND DESCRIBED IN 19th Century By VON MARTIUS.
  • Advisor : EUDES DA SILVA VELOZO
  • COMMITTEE MEMBERS :
  • LAZARO BENEDITO DA SILVA
  • MARA ZELIA DE ALMEIDA
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Sep 18, 2019


  • Show Abstract
  • In Brazil, the use of medicinal plants originated in indigenous culture. Von Martius made great contributions to the knowledge of the Brazilian flora through the Caatinga, Atlantic Forest and Cerrado biomes in the state of Bahia. Objective: To conduct a bibliographic survey of the plants described in Bahia by Von Martius and their therapeutic uses in the early nineteenth century, especially those of Estrada Real stretch Ba. Methodology: quantitative, descriptive, documentary and bibliographical research. .The works studied: "Systema of Brazilian Vegetable Medical Matter", and "Across Bahia: excerpts from Reise in Brasilien / Von Spix and Von Martius." Specielink and GBIF sites were used to confirm the registration of species in herbariums and municipalities in the state of Bahia. To confirm the therapeutic indications of the plants were searched in the databases: PubMed, LILACS, Scielo, Library and Portal CAPES, Revista Fitos, Google Scholars. Results: forty municipalities in Bahia and 47 plants belonging to 26 botanical families were screened. Of these species 74% claimed therapeutic properties. Considering the medicinal uses in current times, 86% have proven therapeutic use. Therapeutic claims were distributed into 13 disease categories according to WHO. In the 19th and 21st century respectively 34% and 21% of plants were used for digestive system pathologies, 21% and 16% for symptoms and signs such as fever and sweat, 13% and 11% for skin diseases. Conclusion: In short, this work enables the encouragement of the valorization of the local flora in the biomes in the state of Bahia, stimulating the use of native medicinal plants by the population

13
  • VALDEENE VIEIRA SANTOS
  • Evaluation of pharmacokinetics and renal penetration of amphotericin B in healthy and Candida albicans infected WISTAR rats using the technique of microdialysis
  • Advisor : FRANCINE JOHANSSON AZEREDO
  • COMMITTEE MEMBERS :
  • ANA LEONOR PARDO CAMPOS GODOY
  • FRANCINE JOHANSSON AZEREDO
  • SANDRA ELISA HAAS
  • Data: Oct 4, 2019


  • Show Abstract
  • Systemic candidiasis is a common opportunistic fungal infection, especially in hospitalized and immunosuppressed patients. Candida albicans is the most common causative agent and most isolated species in these infections, and amphotericin B (AmB) is used for its treatment. For it to have the expected action, it must be able to access its site of action in sufficient concentrations and in its free form, and that the infected tissue does not influence tissue distribution. Therefore, the objectives of this study were to evaluate the pharmacokinetics and renal penetration of AmB by microdialysis (MD) in healthy Wistar rats infected with C. albicans. For this, an analytical and bioanalytical method for the quantification of AmB in microdialysis and plasma of rats was validated. The method was validated using High Performance Liquid Chromatography coupled with a diode array detector (HPLC/ DAD). The mobile phase consisted of acetate buffer (pH = 4) and gradient acetonitrile, effluent with a flow rate of 1.0 mL.min -1, and AmB detection was performed at 407 nm. Relative recovery of AmB by the MD probe was evaluated in vitro, with choice of perfusion flow and analysis of the influence of AmB concentration by the dialysis and retrodialysis method, and in vivo by retrodialysis. The pharmacokinetic and renal penetration study was performed after administration of 1 mg/kg (n = 10) and 2.5 mg/kg (n = 12) of AmB in healthy and C. albicans infected rats, blood and microdialysis samples at predetermined times. The quantification methods are selective and linear in the working range, with adequate precision and accuracy. The perfusion flowrate of the MD probes chosen was 1.5 μL/min, with a relative recovery of 22.5 ± 8.5% and 20.0 ± 6.0% for dialysis and retrodialysis, respectively. Relative recovery of the different concentrations did not show significant difference between dialysis and retrodialysis. Renal in vivo recovery was 10.9 ± 3.7%. After non-compartmental analysis of both plasmatic and renal concentrations and population modeling of plasma data obtained, it can be concluded that C. albicans infection was statistically significant for parameter Vd, and tissue penetration was different between groups (1.6 and 1.1, healthy and infected rats, respectively), with lower tissue distribution of AmB in the infected animals.

14
  • RAFAEL LEONNE CRUZ DE JESUS
  • THERAPEUTIC POTENTIAL OF THE TOPICAL USE OF MENTOL CONTAINING HYDROGEL FOR TREATMENT OF ERECTIBLE HYPERTENSION

  • Advisor : DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • COMMITTEE MEMBERS :
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • FRANCINE JOHANSSON AZEREDO
  • KYAN JAMES ALLAHDADI
  • Data: Oct 23, 2019


  • Show Abstract
  • Introduction: Currently, erectile dysfunction (ED) has been considered a strong marker of cardiovascular disease, including systemic arterial hypertension. In addition, changes such as endothelial dysfunction, the RhoA-ROCK pathway, and the expression of TRP channels in the vasculature are closely linked to hypertension and ED. Objective: To evaluate the influence of chronic topical treatment with gel containing menthol (TRPM8 agonist) on erectile dysfunction symptoms already established in spontaneously hypertensive rats (SHR), as well as on their normotensive controls (wistar). Methods: To evaluate the activity of the TRPM8 channel, strips of corpora cavernosa (CC) isolated from normotensive rats were used. In another set of experiments, SHR and their normotensive wistar controls, both at 10 weeks, were treated topically with menthol-containing hydrogel (experimental group - carbopol 940 + menthol 0.1%) or vehicle (control group - carbopol 940) for 30 days. After treatment, CC and iliac artery (IL) were isolated to subsequent studies of contractility. All experimental protocols were approved by Committee on Ethics in Animal Use - ICS/ UFBA (130/2017). Results: TRPM8 channels are expressed in CC and cumulative administration of menthol (10-7 to 10-3M) induced relaxation in CC strips pre-contracted with phenylephrine wistar animals, (10μM) (Emax = 88.6 ± 3 .02; n = 6). Topical administration of menthol on the penis of animals during 30 days reduced the mean arterial pressure (MAP) of SHR (mmHg = 156.0 ± 6.2, n = 6) when compared to control SHR (mmHg = 191.8 ± 10 .0, n = 5). Reactivity of CC to phenylephrine (10-10 to 10-4M), menthol treatment in SHR animals (Emax = 168.5 ± 7.08 and pD2 = 5.923 ± 0.03; n = 12) increased sensitivity for phenylephrine (Phe) (p <0.01) when compared to control SHR (Emax = 155.1 ± 7.5 and pD2 = 5.783 ± 0.03; n = 9), however, treatment did not promote changes in Phe contractions in CC of wistar animals when compared to control wistar. After treatment, menthol SHR (Emax = 123.0 ± 7.3 and pD2 = 5.609 ± 0.05; n = 8) animals presented lower sensitivity for NE (p <0.05) when compared to control SHR. (Emax = 128.7 ± 9.5; n = 4 and pD2 = 5.699 ± 0.10; n = 4). Additionally, CC reactivity to acetylcholine or SNP from SHR menthol animals was not altered when compared to control. However, in wistar menthol, the curve was shifted to the left when compared to control wistar. The vascular reactivity of the iliac arteries to phenylephrine (10-10 to 3x10-5M) was significantly increased (p<0.01) in menthol SHR rats (Emax = 151.7 ± 8.0 and pD2 = 6.489 ± 0.04; n = 8), with no change in maximal response when compared to SHR control animals (Emax = 163.8 ± 12.2 and pD2 = 6.311 ± 0.04; n = 7), however, the treatment did not cause significant changes. in reactivity to NE. Sensitivity to SNP was significantly increased (p <0.001) in menthol SHR rats (Emax = 109.7 ± 5.9 and pD2 = 7.799 ± 0.06; n = 8), with no change in maximal response when compared to SHR control (Emax = 99.9 ± 1.9 and pD2 = 7.371 ± 0.05; n = 7). However, SNP-induced relaxation in wistar animals (Emax = 101.1 ± 1.4 and pD2 = 8.035 ± 0.06; n = 8) treated with menthol was similar to their wistar controls (Emax = 101.9 ± 2.7 and pD2 = 8.004 ± 0.05; n = 8). Vascular reactivity of the mesenteric arteries to Phe (10-10 to 3x10-5M) did not differ between menthol SHR (Emax = 170.1 ± 6.8 and pD2 = 6.033 ± 0.02; n = 9) and control SHR ( Emax = 162.6 ± 6.1 and pD2 = 5.949 ± 0.02; n = 10). However, in wistar menthol animals (Emax = 127.8 ± 6.0 and pD2 = 6.096 ± 0.02; n = 9), sensitivity to Phe was significantly decreased (p <0.001) when compared to control wistar ( Emax = 129.7 ± 6.2 and pD2 = 6.396 ± 0.03 (n = 11). The treatment also did not promote alterations in NE-induced contractions in the mesenteric arteries of the menthol SHR (Emax = 174.4 ± 5.4 and pD2 = 5.778 ± 0.03; n = 9) and the control SHR (Emax = 181.5 ± 10.7 and pD2 = 5.842 ± 0.04; n = 10). However, in wistar menthol (Emax = 125.6 ± 22.1 and pD2 = 5.839 ± 0.08; n = 8), sensitivity to NE was decreased (p <0.05), with no significant change in maximal response when compared to control wistar (Emax = 123.0 ± 21.7 and pD2 = 6.150 ± 0.13; n = 8). Menthol also increased the sensitivity (p <0.05) of the mesenteric arteries in SHR rats (Emax = 94.3 ± 4.1 and pD2 = 7.640 ± 0.09; n = 5) for acetylcholine when compared to control SHR (Emax = 75.9 ± 5.2 and pD2 = 7.185 ± 0.09; n = 5), with no change among wistar. After treatment, SNP-induced endothelium-independent vasorelaxation in mesenteric arteries did not differ between menthol SHR (Emax = 103.7 ± 2.0 and pD2 = 7.611 ± 0.07; n = 7) and control SHR (Emax = 101.5 ± 2.8 and pD2 = 7.350 ± 0.09; n = 9), however, in wistar menthol animals (Emax = 104.5 ± 5.6 and pD2 = 8.640 ± 0.07; n = 8 ), sensitivity to SNP was significantly increased (p <0.05), with no significant change in maximal response when compared to control wistar (Emax = 108.5 ± 4.0 and pD2 = 8.340 ± 0.05; n = 8 ). Conclusion: We demonstrated the expression of the TRPM8 channel in the corpora cavernosa and that its activation is capable of promoting a relaxing effect of the smooth muscle of the corpora cavernosa of wistar rats and, furthermore, menthol treatment was able to partially improve the responsiveness of corpora cavernosa and vascular tissues of SHR animals thus seem to have therapeutic benefits in cases of erectile dysfunction associated with hypertension when applied topically.

15
  • SUELLEN GONÇALVES DA SILVA
  • Screning of Leishmania major Pteridine Reductase 1 (LmPTR1) inhibitors

  • Advisor : MARCELO SANTOS CASTILHO
  • COMMITTEE MEMBERS :
  • MARCELO SANTOS CASTILHO
  • MAURICIO MORAES VICTOR
  • FRANCO HENRIQUE ANDRADE LEITE
  • Data: Nov 13, 2019


  • Show Abstract
  • Leishmaniasis is a protozoosis that presents itself as a public health challenge in many parts of the world. However, drugs available for the treatment of patients with this disease have a low therapeutic index, relatively long treatment period and reduced therapeutic adherence, which may favor the selection of resistant strains. Consequently, the development of new drugs becomes crucial. Although inhibition of the folate pathway has led to drug discovery against numerous diseases, inhibitors of the enzyme dihydrofolate reductase thymidylate synthase (DHFR-TS) are inefficient against Leishmania spp. The low susceptibility of parasites is associated with Pteridine Reductase 1 (PTR1), an enzyme that works as an alternative pathway for the reduction of folic acid and unconjugated pteridines when DHFR-TS is inhibited. For this reason, inhibitors of this enzyme are promising compounds for the development of new drugs against leishmaniasis. In this context, the aim of this study is to identify molecular fragments and / or lead-like compounds that bind to Leishmania major PTR1 (LmPTR1). Thermal displacement assays showed that the evaluated thiazolidine-2,4-dione derivatives were inactive and that an imidazopyrazine derivative interact with the target in a concentration-response manner. Finally, 42 fragments increase the thermal stability of LmPTR1 (ΔTm> 1.5 ° C). Assays with the covalently fluorescein-linked protein suggest the most affinity fragments for LmPTR1 are LBEA0418 (Kd = 11 ± 1.31mM), LBEA0126 (Kd = 11 ± 1.22mM), LBEA0394 (Kd = 7.5 ± 1.14). ) mM, LBEA0111 (Kd = 1.35 ± 2.11 mM) and LBEA0138 (Kd = 6.5 ± 1.46 mM), while the fragments with the highest binding efficiency are LBEA0111 (LE = 0.33 kcal / mol ) LBEA0447 (LE = 0.32 kcal / mol). Additionally, these assays suggest that six fragments (LBEA0463, 0414, 0418, 0405, 0472, 0473, 0401) interact incompetently with the enzyme cofactor, while seven non-competitively interact (LBEA0453, 0126, 0394, 0111, 0138 , 0447, 0125). Two fragments interact competitively with the substrate (LBEA0418 and LBEA0473), one interacting incompetently (LBEA0463) and two non-competitively (LBE0126, 0394). Because most fragments do not compete with the cofactor or substrate, FTMap and Hotspot Fragment Maps (FragMap) servers were used to identify likely hotspots for these fragments. This information was used to guide molecular coupling studies that will be used to develop leader compounds from fragments.

16
  • ADRIANO DE SOUZA SANTOS MONTEIRO
  • PROPHYLATIC ANTIBIOTIC THERAPY AND INTESTINAL COLONIZATION BY ENTEROBACTERIA (Enterobacteriaceae) IN CHILDREN WITH SICKLE CELL DISEASE
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • ELISANGELA VITORIA ADORNO
  • FABIO DAVID COUTO
  • JOICE NEVES REIS PEDREIRA
  • Data: Nov 22, 2019


  • Show Abstract
  • Infections are the major cause of morbidity and mortality in children with sickle cell disease (SCD) and prophylactic antibiotic therapy (ATB) has shown success in elevating the quality of life of these people. However, the impact of ATB in enterobacteria of the intestinal microbiota in children with SCD has not yet been elucidated. The aim of this study was to evaluate the intestinal enterobacteria composition of children with SCD on prophylactic ATB. Thirty faecal swabs from children with SCD on ATB and 21 swabs from participants without ATB were analyzed. Enterobacteria were isolated in standard procedures, identification and antibiogram were performed by automated methods, and the identification of resistance genes was performed by multiplex PCR. Eleven different species of enterobacteria were found in the entire population studied: seven in the group of children with SCD, on ATB, and nine in the group without ATB, with five common species in both groups. Five species were resistant to antimicrobials, and 52.94% of the E. coli isolates in the SCD group on ATB were resistant to at least one antibiotic (p <0.0001). ESBL production was found in E. coli (4/6) and K. pneumoniae (2/6) in children with SCD, and was associated with benzathine penicillin G (p <0.0003). The β-lactamases TEM (28/37) and CTX-M-gp1 (3/37) were the most predominant, followed by CTX-M-gp9 (2/37), CTX-M-gp2 (1/37) and SHV (1/37).  Despite the presence of antibiotic resistant enterobacteria, especially E. coli, prophylactic ATB in children with SCD is essential, especially to reduce deaths caused by infections.
17
  • ANA PAULA DÓRIA DE ARAÚJO CRUZ
  • APPLICATION OF SEED OIL OF Passiflora cincinnata IN EMULSIONED SYSTEMS AND EVALUATION OF ANTI-INFLAMMATORY ACTIVITY IN ARTHRITIS MODEL IN MICE
  • Advisor : DENIS DE MELO SOARES
  • COMMITTEE MEMBERS :
  • DENIS DE MELO SOARES
  • HENRIQUE RODRIGUES MARCELINO
  • NATHALIE RIBEIRO WINGERT
  • Data: Dec 13, 2019


  • Show Abstract
  • The use of natural products has spread as complementary alternatives to conventional therapies. This choice is linked to the phytochemical components that gives it functional properties used as a health benefit. Therefore, the oil from the seeds of Passiflora cincinnata, a species of semi-arid passion fruit, is made up of a range of phytoconstituents, which may cite its high content of polyunsaturated fatty acids which are responsible for several therapeutic properties that contribute to the body's homeostasis and treatment of various conditions. The aim of this study was to elaborate a topical product using P. cincinnata oil incorporated to an O / A emulsion by cold emulsification, in order to allow emollience, hydration, restoration of the epidermis and release of the herbal constituent. The cosmetic vehicle chosen was a self-emulsifying Hydrafresh®, which detects its phospholipid composition, which contributes to the efficiency of the final product. Oil extraction was performed by Soxhlet technique following thermogravimetric analysis (TG), formulation preparation, stability, toxicity and pharmacology tests, in order to demonstrate the viability of the topical product with potential therapeutic effect. Of mucosal irritation by HET-CAM test and experiment to evaluate potential anti-inflammatory activity in mouse knee arthritis model. Therefore, the evaluation of mucosal irritation by the HET-CAM test and experiment to evaluate the potential anti-inflammatory activity in a mouse knee arthritis model were the toxicity and pharmacological tests adopted. The crude oil yield of P.cincinnata was (13.07%), with thermal stability up to 426°C. Regarding the results found in the physicochemical stability tests, the formulations were approved, maintaining adequate organoleptic characteristics, with a pH range between 5,8-6,6. In the analysis of the diameter of its droplets, the HYOC10 formulation exhibited, in due order, Dv (50) (10,65 ± 0.12μm) and Dv (90) (28,0 ± 0,15μm), showing stastitic difference between HYOCON and HYOC, which had sizes with Dv (50) (3,04 ± 0,04μm), (3,32 ± 0,07μm) and Dv (90) (9,7 ± 0,57μm), (11,09 ± 0,83μm), however, all were identified as macroemulsions. HYOC10 showed an in vitro spreadability increase after accelerated stability with Ei max of (1764,55 ± 445,35mm2 ), reaffirming this characteristic in the analysis of viscosity and texture. In the HET-CAM test both the isolated oil and the HYOC10 obtained on the punctuated scale, respectively the values of (4,8 ± 0,5) and (4,4 ± 0,8) that classified them as weak irritants. In the pharmacological test, groups were initially tested with P.cincinnata oil, it was noticed that group D10, which corresponded to a concentration of 10%, was the one that responded best in controlling inflammation, knowing this, the formulations HYCON, HYOC10 were also further tested, where they responded positively in reducing joint edema associated with decreased neutrophils. Therefore, the present work resulted in an innovative and promising topical product from natural sources, with a potential anti-inflammatory activity aiming at its use in the pharmacological treatment of clinical manifestations related to arthritis.

2018
Dissertations
1
  • MAIARA DE SOUZA OLIVEIRA
  • CYTOTOXIC ACTIVITY OF A NEW PLATINUM COMPLEX CONTAINING PIPLARTIN AS LIGENT
  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • DANIEL PEREIRA BEZERRA
  • CAROLINE BRANDI SCHLAEPFER SALES
  • JUNIA RAQUEL DUTRA FERREIRA
  • Data: Jan 15, 2018


  • Show Abstract
  • Piplartine, also known as piperlongumine, is an alkaloid/amide found in plants of the genus Piper, which has been extensively studied for its cytotoxic properties. In addition, metal complexes, such as platinum complexes, are successfully used in the treatment of cancer. Thus, the objective of the present work was to study the cytotoxic properties of a new platinum complex containing piplartine as a binder (CPP01). The platinum complex CPP01 was synthesized and tested against different tumor cell types (HepG2, HL-60, HCT116, SCC9, HSC3, MCF7, K562 and B16-F10) and non-tumor (MCR5 and PBMC) cells by the alamar assay blue after 72 h of incubation. Subsequently, HL-60 cells were incubated for 24 and 48 h with CPP01 (1, 2 and 4 μM) and the number of viable cells was determined by the trypan blue exclusion assay. Cell morphology was assessed after staining with may-grunwald-giemsa and cell cycle analysis, mitochondrial transmembrane potential and labeling for annexin V/propidium iodide, reactive oxygen species were determined by flow cytometry. The activity of caspase-3 was determined by colorimetric assay, and DNA intercalation assay was performed by fluorescence method. The complex CPP01 showed IC50 values for tumor cells ranging from 0.95 to 6.82 μM for the HSC3 and MCF7 lines, which presented an IC50 value of 7.45 and 9.18 μM for non-tumor cells MCR5 and PBMC, respectively. Piplartine showed IC50 values for tumor cells ranging from 5.39 to 18.58 μM for the HCT116 and K562 lines, which presented an IC50 value of 17.34 and 34.22 μM for non-tumor cells MCR5 and PBMC, respectively. In trypan blue exclusion analysis, the complex CPP01 reduced the number of viable cells without increasing the number ofnon-viable cells. Typical morphology of apoptotic cell death, an increase in internucleosomal DNA fragmentation, reduction of mitochondrial transmembrane potential, induces oxidative stress and a marked labeling for annexin V and caspase-3 were observed complex-treated HL-60 cells, suggesting induction of apoptotic cell death. In conclusion, it is possible to verify that the complex CPP01 showed more potent cytotoxic activity than piplartine in different tumor cell lines, inducing oxidative stress and apoptotic cell death mediated by caspases in HL-60 cells.

2
  • MAIARA DE SOUZA OLIVEIRA
  • CYTOTOXIC ACTIVITY OF A NEW PLATINUM COMPLEX CONTAINING PIPLARTIN AS LIGENT
  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • DANIEL PEREIRA BEZERRA
  • CAROLINE BRANDI SCHLAEPFER SALES
  • JUNIA RAQUEL DUTRA FERREIRA
  • Data: Jan 15, 2018


  • Show Abstract
  • Piplartine, also known as piperlongumine, is an alkaloid/amide found in plants of the genus Piper, which has been extensively studied for its cytotoxic properties. In addition, metal complexes, such as platinum complexes, are successfully used in the treatment of cancer. Thus, the objective of the present work was to study the cytotoxic properties of a new platinum complex containing piplartine as a binder (CPP01). The platinum complex CPP01 was synthesized and tested against different tumor cell types (HepG2, HL-60, HCT116, SCC9, HSC3, MCF7, K562 and B16-F10) and non-tumor (MCR5 and PBMC) cells by the alamar assay blue after 72 h of incubation. Subsequently, HL-60 cells were incubated for 24 and 48 h with CPP01 (1, 2 and 4 μM) and the number of viable cells was determined by the trypan blue exclusion assay. Cell morphology was assessed after staining with may-grunwald-giemsa and cell cycle analysis, mitochondrial transmembrane potential and labeling for annexin V/propidium iodide, reactive oxygen species were determined by flow cytometry. The activity of caspase-3 was determined by colorimetric assay, and DNA intercalation assay was performed by fluorescence method. The complex CPP01 showed IC50 values for tumor cells ranging from 0.95 to 6.82 μM for the HSC3 and MCF7 lines, which presented an IC50 value of 7.45 and 9.18 μM for non-tumor cells MCR5 and PBMC, respectively. Piplartine showed IC50 values for tumor cells ranging from 5.39 to 18.58 μM for the HCT116 and K562 lines, which presented an IC50 value of 17.34 and 34.22 μM for non-tumor cells MCR5 and PBMC, respectively. In trypan blue exclusion analysis, the complex CPP01 reduced the number of viable cells without increasing the number ofnon-viable cells. Typical morphology of apoptotic cell death, an increase in internucleosomal DNA fragmentation, reduction of mitochondrial transmembrane potential, induces oxidative stress and a marked labeling for annexin V and caspase-3 were observed complex-treated HL-60 cells, suggesting induction of apoptotic cell death. In conclusion, it is possible to verify that the complex CPP01 showed more potent cytotoxic activity than piplartine in different tumor cell lines, inducing oxidative stress and apoptotic cell death mediated by caspases in HL-60 cells.

3
  • Viviane de Sousa Moreira Almeida
  • Antimicrobial resistance in microbiota of the oral cavity
  • Advisor : JOICE NEVES REIS PEDREIRA
  • COMMITTEE MEMBERS :
  • JOICE NEVES REIS PEDREIRA
  • JAILTON DE AZEVEDO SILVA JUNIOR
  • PAULO JOSE LIMA JUIZ
  • Data: Feb 19, 2018


  • Show Abstract
  • Antimicrobial resistance may make it difficult to treat important diseases and the oral cavity plays a role in the spread of antibiotic resistant bacteria. The objective of this study was to evaluate the prevalence of resistance genes in the oral microbiota and its relation with caries and periodontal disease. The study was observational and prospective, characterized as cross-sectional type and was performed with individuals who sought care at the Bahia Dental School (FOUFBA) and the Faculty of Pharmacy of the Federal University of Bahia (LACFAR-UFBA), between October to November 2016. Were included 110 people of both sexes, over 18 years old, who agreed to participate in the study with a clinical examination, were interviewed through questionnaires containing personal and epidemiological data. A clinical examination of the volunteers was done and saliva and plaque collection were performed, which were stored in sterile collectors to be aliquoted and frozen at -70 ° C. Bacterial DNA was extracted from saliva and plaque samples using the Maxwell system ( Promega) and PCR were performed to identify the resistance genes: aac(6´),blaNDM, blaSPM, blaOXA-48, CTX-M, blaKPC, blaIMP, blaVIM, pbp-2b, mecA, erm(A), erm(B) e erm(C), blaTEM, blaSHV e nim. The program Epi-Ifo Windows v.3.5.3 (CDC, 2011) was used for storage and analysis of epidemiological data. The erm(B) gene was found in 58.2% (64/110), the blaTEM gene in 16.4% (18/110), the aac (6 ') gene in 1.8% (2/110), the mecA gene at 2.7% (3/110) and the pbp-2b gene at 1.8% (2/110). Resistance genes had a very similar prevalence in participants with caries and without caries, and in individuals with periodontal disease, they were more prevalent, especially in PSR score 2. However, it was not possible to establish a relationship between the presence of dental caries and the periodontal status of the study participants and the presence of resistance genes.

4
  • SILVIA SOUZA DE CARVALHO
  • Evaluation of the reactivity of specific IgG and IgA antibodies and nitric oxide levels in day care children infected with Giardia duodenalis
  • Advisor : MARCIA CRISTINA AQUINO TEIXEIRA
  • COMMITTEE MEMBERS :
  • MARCIA CRISTINA AQUINO TEIXEIRA
  • MARIA LUIZA BRITO DE SOUSA ATTA
  • RICARDO WAGNER DIAS PORTELA
  • Data: Feb 22, 2018


  • Show Abstract
  • Giardia duodenalis infection is common in poor countries and affects mainly children, with the majority of cases being asymptomatic. The trophozoites of Giardia duodenalis colonize the surface of the intestinal mucosa, stimulating local and systemic immune responses through their antigens. The infection induces the production of specific antibodies that can play a role in the elimination of the parasite, as well as being useful as a diagnostic tool to monitor the exposure to the infection. In addition to the humoral immune response, some mediators of the innate immune response, such as nitric oxide (NOx), may also be involved in protection against giardiasis. The main objective of this study was to analyze the reactivity of specific IgG and IgA antibodies and nitric oxide levels in children infected with Giardia duodenalis. This study evaluated 187 samples of serum, feces and saliva from children attending two daycare centers in the city of Salvador, Bahia. The fecal samples were examined by sedimentation by centrifugation to investigate enteroparasites, and by centrifugal fecal flotation in zinc sulfate method and coproantigen screening for diagnosis of G. duodenalis. The reactivity of seric IgG and IgA anti-Giardia antibodies was evaluated by an in-house immunoenzymatic method (ELISA) and the concentration of nitric oxide in serum and saliva were measured by the Griess method. The frequency of Giardia duodenalis infection in children was 8.5% (16/187); 12 cases were diagnosed by microscopy and coproantigens, and four cases were exclusively by the second, with high agreement between the methods (kappa = 0.84). The sensitivity and specificities of ELISAs were 80.0% and 90.0% for IgG, and 80.0% and 83.3% for IgA. Respectively, the reactivity of specific antibodies in the sera tested was 16% (30/187) for IgG and 28.9% (54/187) for IgA. No statistically significant differences were found in the nitric oxide dosagebetween groups, although a higher proportion of samples with high NOx concentrations (above 41μmol/L) were observed in both serum (25.0%) and saliva (33.3%) of children infected with Giardia duodenalis. Over 70% of the children were classified as eutrophic, with no differences between the Body Mass Index (BMI) and the parasitological positivity, or the reactivity of anti-Giardia duodenalis antibody. The high rate of anti-Giardia duodenalis IgG and IgA antibodies shows the high endemicity and early exposure to this protozoan in children, which may be useful as an auxiliary diagnostic tool. Due to limited sample size of parasitized children in this study, there was not possible to infer the relationship of serum and secretory NO levels with Giardia duodenalis infection.

5
  • TALITA NUNES DOURADO CARVALHO

  • EVALUATION OF ANTI-RODS AND ANTI-RINGS (ANTI-RR) ANTIBODIES IN PEOPLE WITH CHRONIC HEPATITIS C, PRE AND POST-TREATMENT

  • Advisor : MARIA LUIZA BRITO DE SOUSA ATTA
  • COMMITTEE MEMBERS :
  • MARCIA CRISTINA AQUINO TEIXEIRA
  • MARIA LUIZA BRITO DE SOUSA ATTA
  • RYAN DOS SANTOS COSTA
  • Data: Feb 28, 2018


  • Show Abstract
  • Hepatitis C virus (HCV) infection is a major cause of liver disease. Anti-rod and antiring (anti-RR) antibodies have been described as autoantibodies to antigenic targets that are key enzymes in nucleic acid and phospholipid biosynthesis, the CTPS1 (cytidina triphosphate synthase 1) and IMPDH2 (inosine monophosphate dehydrogenase 2). These autoantibodies have been associated with non-responders or relapses to treatment with interferon-α plus ribavirin (IFN/RBV). OBJECTIVES: To investigate the presence and frequency of anti-RR antibodies in patients with chronic hepatitis C (CHC), pre and post treatment, its relation to other autoimmunity markers, and associations with clinical and virological findings. PATIENTS, MATERIALS AND METHODS: Retrospective cohort study, including serum samples from 52 male and female individuals from the HUPES Hepatitis Clinic, clinically and serologically diagnosed for hepatitis C. The data from genotyping and staging of fibrosis in patients with CHC were obtained from medical records. The test of anti-RR antibodies was performed by indirect immunofluorescence (IIF) using as antigens Hep-2 cells, obtained from a commercial set of immunodiagnostic (INOVA Diagnostics). The autoimmunity markers, rheumatoid factor and cryoglobulins were investigated by nephelometry and cryoprecipitation, respectively. RESULTS: The prevalence of antiRR antibodies was 2% in untreated patients, 4% in the 12th week of treatment and 26% in the 24th week of treatment. No association was found between the presence of antiRR antibodies and clinical and virological parameters of the infection, but genotype 3 was associated with anti-RR in the 24th week of antiviral treatment. CONCLUSION: The presence of anti-RR antibodies is related to the duration of antiviral treatment with interferon-α plus ribavirin in hepatitis C.

6
  • VANESSA COSTA DOS SANTOS
  • RESPOSTA IMUNE ADAPTATIVA DE SUBCLASSES DE ANTICORPOS IgG
    NA HEPATITE C CRÔNICA

  • Advisor : AJAX MERCES ATTA
  • COMMITTEE MEMBERS :
  • AJAX MERCES ATTA
  • ANA LEONOR PARDO CAMPOS GODOY
  • ROBSON DA PAIXAO DE SOUZA
  • Data: May 16, 2018


  • Show Abstract
  • Introduction: Chronic hepatitis C virus infection affects millions of people living in different countries, including Brazil. Objective: To investigate the adaptive immune response mediated by subclasses of IgG antibodies to the recombinant core and NS3 antigens in patients with chronic hepatitis C. Methods: Sixty patients chronically infected with HCV genotype 1, without antiviral treatment, and 60 healthy subjects participated in the study. Serum levels of alanine aminotransferase, HCV viremia, the presence of cryoglobulinemia and liver fibrosis were determined. The presence of serum IgG1 and IgG4 antibodies against recombinant HCV NS3 core and non-structural protein antigens was investigated by indirect ELISA. Results: Anti-core and anti-NS3 IgG1 antibodies were detected in 33/60 (55%) and 46/60 (77%) of the patients, whereas two healthy controls reacted with one of the antigens (NS3). Anti-core IgG4 antibodies were not detected in both groups, while 30/60 (50%) of the patients had anti-NS3 IgG4 antibodies. Except for the observation of higher levels of anti-NS3 IgG4 antibodies in patients with low viremia (<8 x 105 IU/mL), IgG1 and IgG4 antibody levels were not influenced by ALT levels, the presence of cryoglobulinemia or degree of fibrosis hepatic. Conclusions: A vigorous production of anti-core and anti-NS3 IgG1 antibodies was found in chronic hepatitis C patients. In contrast, IgG4 antibodies seemed to be only produced to the NS3 non-structural antigen and appeared to participate in viremia control.

7
  • LARISSA MENDES BOMFIM
  • ANTINEOPLASTIC POTENTIAL OF NEW RUTENUM COMPLEX WITH 6-METHYL-2-THIOURACIL
  • Advisor : DANIEL PEREIRA BEZERRA
  • COMMITTEE MEMBERS :
  • DANIEL PEREIRA BEZERRA
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • ROSANE BORGES DIAS
  • Data: May 28, 2018


  • Show Abstract
  • Ruthenium compounds have gained great interest because of their potent cytotoxicity in cancer cells, however, many of their potential applications remain unexplored. In this work, we investigated the antineoplastic potential of two new ruthenium complexes with 2-thiouracil (CR01 and CR02) in different cell models and their mechanisms of action in HL-60 human promyelocytic leukemia cells. The cytotoxicity of the complexes against different neoplastic lines was determined by the alamar blue assay. Subsequently, HL-60 human promyelocytic leukemia cells were incubated with CR01 and CR02 at concentrations of 1 and 2μM, and the number of viable cells was determined by the trypan blue exclusion assay. Cell cycle analysis, mitochondrial transmembrane potential and labeling for annexin V / propidium iodide were determined by flow cytometry. The caspase activation assay was also performed to evaluate the apoptotic processes induced by the complexes. The in vivo antitumor activity of the CR01 complex was evaluated in C.B-17 SCID mice inoculated with HL-60 cells. We found that CR01 and CR02 exhibited a potent cytotoxic effect on a panel of cancer cell lines. HL-60 cells treated with the complexes showed a reduction in the number of viable cells, increased fragmentation of internucleosal DNA, depolarization of mitochondrial transmembrane potential, activation of caspases 3, 8 and 9 and increased phosphatidylserine externalization, indicating induction of cell death apoptosis mediated by caspases. In addition, pretreatment of HL-60 cells with inhibitors of the major families of the MAPK signaling pathway (inhibitors of p38, JNK / SAPK and ERK1 / 2) prevented CR01 and CR02 induced apoptosis. In the in vivo model, CR01 inhibited the development of HL-60 cells in C.B-17 SCID mice, exhibiting a potent antitumor effect in vivo. These results indicate that ruthenium complexes with 2-thiouracil have anticancer potential under different carcinogenic strains, and induce caspase-mediated apoptotic cell death in HL-60 cells. In addition, the CR01 complex was able to inhibit its development in vivo.

8
  • ROUSILANDIA DE ARAUJO SILVA
  • Development of formulations for cutaneous moisturizing by cold emulsification: evaluation of sensorial properties

  • Advisor : NEILA DE PAULA PEREIRA
  • COMMITTEE MEMBERS :
  • ADEMIR EVANGELISTA DO VALE
  • ANDRE LUIS MORAIS RUELA
  • NEILA DE PAULA PEREIRA
  • Data: May 30, 2018


  • Show Abstract
  • Skin products occupy the eighth position in the ranking of the most consumed cosmetics, and moisturizers represent the most important class of this group, since they play an important role in the prevention of xerosis and skin aging. Emulsion is the most used pharmaceutical form for topical products and their preparation requires energy, time and equipment expenditures, so the cold emulsification process minimizes the costs of the production process. In this context, the present study aims to develop cold emulsified formulations with the aid of the Gelaid CPE CF ® silicon base using vegetable oils of Sweet Almond (GA), Gueroba (GGE) and Macaúba (GM) and Urea (GU) moisturizing active ingredients. Oleate Lactate (GL), Octadecila Lactate (GO) and Urea Glycospheres (GG) evaluating their sensory properties. The study was divided into three stages. The first one was the study of the Stability of the Formulations following its behavior during the 90 days by means of the macroscopic evaluation, analysis of the pH, Spreadability in vitro. All formulations remained stable without change in their organoleptic characteristics and remained within the cutaneous pH. In the group of vegetable oils the GM formulation (Eimax 3739.8mm2) presented better spreadability performance and in the group with active moisturizing the GO formulation (Eimax 2687.95 mm2). The second stage was the sensorial characterization through the determination of the texture profile and the rheological behavior. In the texture, the GGE formulation recorded lower values of firmness (4.73 g) and in the group with active the GO formulation (4.63 g). The rheological evaluation determined that all formulations presented pseudoplastic behavior with thixotropy and better recovery for GM and GO formulation in their respective groups. The last step of the study was the sensory analysis by the preference method in which the formulations with the highest score, in the case of GGE oils (44 points) and the active ones at GO (60 points) were considered preferred as regards spreadability by the research participants. The results found in the formulations with oil and moisturizing active demonstrate the feasibility in developing moisturizing products by the process of cold emulsification with different raw materials.

9
  • ROUSILANDIA DE ARAUJO SILVA
  • Development of formulations for cutaneous moisturizing by cold emulsification: evaluation of sensorial properties

  • Advisor : NEILA DE PAULA PEREIRA
  • COMMITTEE MEMBERS :
  • ADEMIR EVANGELISTA DO VALE
  • ANDRE LUIS MORAIS RUELA
  • NEILA DE PAULA PEREIRA
  • Data: May 30, 2018


  • Show Abstract
  • Skin products occupy the eighth position in the ranking of the most consumed cosmetics, and moisturizers represent the most important class of this group, since they play an important role in the prevention of xerosis and skin aging. Emulsion is the most used pharmaceutical form for topical products and their preparation requires energy, time and equipment expenditures, so the cold emulsification process minimizes the costs of the production process. In this context, the present study aims to develop cold emulsified formulations with the aid of the Gelaid CPE CF ® silicon base using vegetable oils of Sweet Almond (GA), Gueroba (GGE) and Macaúba (GM) and Urea (GU) moisturizing active ingredients. Oleate Lactate (GL), Octadecila Lactate (GO) and Urea Glycospheres (GG) evaluating their sensory properties. The study was divided into three stages. The first one was the study of the Stability of the Formulations following its behavior during the 90 days by means of the macroscopic evaluation, analysis of the pH, Spreadability in vitro. All formulations remained stable without change in their organoleptic characteristics and remained within the cutaneous pH. In the group of vegetable oils the GM formulation (Eimax 3739.8mm2) presented better spreadability performance and in the group with active moisturizing the GO formulation (Eimax 2687.95 mm2). The second stage was the sensorial characterization through the determination of the texture profile and the rheological behavior. In the texture, the GGE formulation recorded lower values of firmness (4.73 g) and in the group with active the GO formulation (4.63 g). The rheological evaluation determined that all formulations presented pseudoplastic behavior with thixotropy and better recovery for GM and GO formulation in their respective groups. The last step of the study was the sensory analysis by the preference method in which the formulations with the highest score, in the case of GGE oils (44 points) and the active ones at GO (60 points) were considered preferred as regards spreadability by the research participants. The results found in the formulations with oil and moisturizing active demonstrate the feasibility in developing moisturizing products by the process of cold emulsification with different raw materials.

10
  • LAURA MARIA SANTOS DO NASCIMENTO
  • APPLICATION OF SEED OIL AND EXTRACT FROM LEAVES
    Psidium guajava L. IN EMULSIFIED SYSTEMS WITH COSMETIC POTENTIAL

  • Advisor : NEILA DE PAULA PEREIRA
  • COMMITTEE MEMBERS :
  • FABIO ROCHA FORMIGA
  • NEILA DE PAULA PEREIRA
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • Data: Jun 12, 2018


  • Show Abstract
  • The guava (Psidium guajava L.) is a tropical fruit of great prominence, with high levels of vitamin C, phenolic compounds, carotenoids, besides having high concentration of vegetable oil, being potential for use in cosmetic formulations as active and/or component of the oil phase of emulsifying systems. In this context, the objective of this work was to obtain O/W emulsions incorporated with fixed oil, obtained from guava seeds, and the hydroethanolic extract of the leaves of P. guajava L., Paluma cultivar, claiming to obtain a cosmetic formulation moisturizing and antioxidant. Initially, the fixed oil of the guava seeds was obtained in a Soxhlet device using hexane, and the derivatized fatty acids were characterized by gas chromatography coupled to mass spectrometry. The physico-chemical properties of the oil were evaluated for relative density, peroxide index, acidity index, saponification index and iodine index. In the second stage the hydroethanolic extract of the leaves of P. guajava was obtained by the maceration method followed by percolation, being evaluated the pH and the relative density of the same. The liquid extract was concentrated by atomization in spray dryer and from the dry extract, the water activity, the quantification of the total phenolic compounds and the antioxidant activity by the free radical sequestration DPPH• were determined. In addition to the hydroethanolic extract and fixed seed oil, the in vitro cytotoxicity assay was performed in murine macrophages of the J774 balb/c lineage, using the colorimetric method of resazurin reduction (Alamar Blue®), in addition to thermogravimetric analysis to evaluate the thermal stability through the TG/DTG curves. Afterwards, the ionic and non-ionic emulsions incorporated with the fixed oil of the seeds were obtained, being submitted to the test of analysis of the organoleptic and microscopic characteristics, pH determination, accelerated stability, in vitro spreadability and texture profile during 90 days. Based on the performance of the formulations, new non-ionic emulsions containing 2.5%, 5.0% and 7.0% of the hydroethanolic extract were prepared. The antioxidant activity was analyzed as a criterion for the development of an emulsion containing 2, 5% of fixed seed oil and hydroethanolic extract of guava leaves. The emulsion containing oil and extract was evaluated for the antioxidant potential. The results showed a predominance of unsaturated fatty acids for the fixed seed oil, with linoleic acid predominantly (80.41%). As for the physico-chemical properties, the oil was in accordance with the literature, making it potentially applicable in the cosmetics industry. As for the extract the relative density was within expected range and the pH was slightly acidic. The water activity of the dry extract was 0.265 (Aw), which does not favor the growth of microorganisms. The total phenolic content was 127.78 ± 2.87 mg gallic acid /g extract. The antioxidant activity of the ethanolic extract was 89.15 ± 0.29% and EC50 of 0.1008 mg/mL, these results being promising for the development of innovative cosmetics. In the thermal stability evaluation the seed oil was stable at a temperature of 360ºC, the leaves extract is stable below 45ºC. The oil and extract were not cytotoxic to the cell line tested until the concentration of 10 µg/mL. The obtained emulsions were stable during the period evaluated, and the pH did not suffer a marked variation. Regarding the spreadability and texture, the non-ionic emulsions presented better performance, where the emulsion containing the fixed oil of the seeds was distinguished by maintaining the behavior during the 90 days. For the emulsions containing the hydroethanolic extract, antioxidant activity was observed, varying from 45.88 ± 1.11 to 64.58 ± 0.93%. The emulsion, increased 2.5% of fixed seed oil and hydroethanolic extract of the leaves presented promising results of antioxidant activity of 54.89 ± 1.53%, opening the possibility for the development of an innovative phytocosmetics with antioxidant and moisturizing activity.

11
  • CÁSSIA VARGAS LORDÊLO
  • FREQUENCY OF C677T, G1691A AND G20210A POLYMORPHISMS IN THE GENES OF METHYLENETETRAHYDROPHOLATE REDUTASE, FACTOR V E PROTROMBINE IN CAROTYDE DISEASE
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • RICARDO DAVID COUTO
  • ROQUE ARAS JUNIOR
  • CYNARA GOMES BARBOSA
  • Data: Jul 25, 2018


  • Show Abstract
  • In recent years, molecular targets are being direct and indirect associated with risk
    markers in the development of cardiovascular diseases. In this context, the study
    objective was to determine the MTHFR enzyme gene polymorphism C677T, Factor V
    gene G1691A and Prothrombin gene G20210A frequency’s, and to investigate the role
    of risk biomarkers in the development of carotid artery disease (CAD). For this purpose,
    DNA extraction was performed from the whole blood samples of 62 patients, militaries,
    and their dependents, from the Hospital Naval de Salvador, Bahia. The collected DNA
    samples were submitted to PCR, followed by enzymatic digestion (RFLP). The
    associations between CAD, biomarkers and genetic polymorphisms were measured by
    Fisher's test. The mean age of participants was 57.85 ± 15.74, ranging from 20 to 77
    years. Among individuals who underwent carotid doppler sonography, 46.8% did not
    show CAD, and 53.2% had some degree of atherosclerosis in the carotid artery. The
    frequency of Factor V G1691A polymorphism in the study population was 1.6%, for
    heterozygous genotype (GA). The G20210A polymorphism in the Prothrombin gene
    showed the frequency of 3.2% of heterozygosity (GA), and the MTHFR enzyme gene
    C677T polymorphism was 24.2% for heterozygotes (CT), and 1.6% for homozygotes
    (TT). The results suggested that there is no association between the MTHFR enzyme
    gene C677T polymorphism and the CAD in the studied population and the Plasma
    Atherogenic Index (AIP) has been shown to be a good parameter for the associated
    diagnostic assistance of subclinical CAD risk, mainly in females.

12
  • PAULA SCHONS VIECELI
  • THERAPEUTIC POTENTIAL OF PHYSALIS ANGULATA EXTRACT RICH IN PHYSALINES IN INDUCED PERIODONTAL DISEASE MODEL
  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CRISTIANE FLORA VILLARREAL
  • FRANCINE JOHANSSON AZEREDO
  • TERCIO CARNEIRO RAMOS
  • Data: Jul 26, 2018


  • Show Abstract
  • Periodontal disease (PD) is a disease with one of the greatest risks for oral health, and triggers tooth loss if left untreated. Fisalines are dry-steroids with anti-inflammatory and immunomodulatory properties. Aiming to establish the scientific basis for herbal medicine future development, this paper evaluated the therapeutic potential of physalis angulata extract rich in fisalines (EPA) in animal model of periodontal disease. PD was induced by injections of LPS 20 μg / 1μl for 28 days. Treatments consisted of daily oral administrations of EPA 50 or 100 mg / kg, vehicle (saline + 5% DMSO) or nimesulide (25 mg / kg, reference drug) during 14 days. The jaws and gingiva were collected to measure bone loss, histology, PCR and ELISA. Stomach, kidneys, liver, heart and blood were collected for toxicity testing. The results showed that EPA (50 and 100 mg / kg) was able to inhibit alveolar bone loss when compared to vehicle (p <0.05). Treatment with EPA (50 and 100 mg / kg) inhibited expression of mRNA and proinflammatory cytokines IL-1β and IL-6 (p <0.05), but not TNF-α. An increase in the anti-inflammatory cytokine TGF-β (p <0.05) was noted, but not in IL-10. EPA reduced the expression of MMP-9, but not TIMP-1, in gingival tissue (p <0.05).The histopathological analysis did not show any changes in the experimental groups. The analysis of systemic toxicity did not report statistically significant differences in blood parameters between groups treated with EPA and vehicle. On the other hand, treatment with nimesulide altered the RBC (p <0.05), HGB (p <0.05), HCT (p <0.05) and MCHC (p <0.05) parameters compared to vehicle. Biochemical parameters have not changed. A histological analysis of the stomach indicated histopathological changes in all treated groups in compered to naive, including presence of ulcer, necrosis and hemorrhage.This study demonstrated that EPA has therapeutic properties in PD and may represent a promising coadjutant approach in the treatment of that disease. On the other hand, the gastric toxicity evidenced needs a better investigation.

13
  • Homegnon Antonin Ferreol Bah

  • Influence of the enzyme delta-aminolevulinic acid dehydratase on the association between plumbemia and children's intellectual function

  • Advisor : JOSE ANTONIO MENEZES FILHO
  • COMMITTEE MEMBERS :
  • JOSE ANTONIO MENEZES FILHO
  • FERNANDO MARTINS CARVALHO
  • EDNA LUCIA SANTOS DE SOUZA
  • Data: Jul 31, 2018


  • Show Abstract
  • Introduction: Lead (Pb) is a ubiquitous environmental contaminant which affects children cognition and inhibits activity of the delta-aminolevulinic acid dehydratase (enzyme ALAD). The community of Maragogipinho, (district of Aratuípe) is exposed to Pb due to the glazed ceramics produced without protective measures.
    Objective: This work aimed to evaluate the association between Pb exposure and the intellectual level of children considering the interaction with the enzyme ALAD.
    Methods: Children of 5.5 to 13 years from two areas (four schools) of Maragogipinho, previously classified as low exposure (LEx; n = 34) and Moderate Exposure (MEx; n = 40) were compared with children (n = 69) of the urban center of the municipality considered as control (CG). The blood lead level (BLL) and Pb loading rate of (RtPb) in settled dust of the children residences (n = 108) were determined. The nutritional status (z-score A / I), hemoglobin concentration (Hb), iron deficiency (Ferritin), activity and polymorphism of the enzyme ALAD were evaluated. The questionnaire HOME was applied, while the non-verbal intelligence of the children and mothers were evaluated using the Raven’s Progressive Matrices.
    Results: The median (range) of RtPb in settled dust from residence house was 65 (6 - 1964) μg Pb / m2 /30 days. The median (range) of BLL was 1.0 (0.1-21.3) μg/dL and was not influenced by the sex and socioeconomic status. The mean (SD) of Hb, ALAD, ferritin, z-score H/A, and raw score of Raven were respectively 13.2 (0.93) g/dL; 71 (31 - 113) U/L; 27.1 (3.1 - 152.8) ng/mL; 0.03 (-2.6 - 4.7) and 19.3 (5.6) while the phenotypes of the enzyme ALAD were ALAD 1/1 (97.9%) and ALAD 1/2 (2.1%). Spearman correlation confirmed the relationship of children BLL with the RtPb in settled dust from their homes (rho = 0.368, p <0.001), inhibition of ALAD enzyme activity by Pb (rho = -0.587; p ˂0.001). The prevalence rates of high blood lead (>5 μg/dL) in the LEx and MEx areas when compared to the CG were 1.5 and 3.7 respectively, indicating a higher risk of children residing in the area near the pottery facilities. No significant association was found between BLL and the Raven score, but its correlation with the activity of the ALAD enzyme (rho = -0.205; p = 0.021; n = 127) suggested the neuroprotective role of that enzyme.
    Conclusion: From this investigation, it can be concluded the existence of moderate exposure of children in this community, confirmed the hematological toxicity of Pb, the role of dust in children contamination. Among the main determinants of children BLL are: living in the vicinity of pottery facilities and involvement in the production of ceramics. The failure to observe the association between BLL and children's nonverbal intelligence, probably due to the lack of sensitivity of the neuropsychological instrument used, suggesting to further the investigation with more adequate cognitive tests.

14
  • MARCOS CUSTÓDIO FIUZA
  • S447X POLYMORPHISM ASSOCIATION IN THE LIPASE ENZYME GENE LIPOPROTEIC WITH CARDIOVASCULAR RISK MARKERS IN DISEASE CAROTIDE
  • Advisor : RICARDO DAVID COUTO
  • COMMITTEE MEMBERS :
  • ANA PAULA CAIRES DOS SANTOS VALVERDE
  • JOICE NEVES REIS PEDREIRA
  • RICARDO DAVID COUTO
  • Data: Aug 3, 2018


  • Show Abstract
  • In recent years, new biomarkers suggestive of cardiovascular risk are being tested for
    the associated diagnosis of atherosclerotic disease. Thus, the objective of this study
    was to identify the presence of LPL enzyme gene S447X (C1595G) polymorphism and
    associate its presence with imaging data and serum biomarkers in patients with and
    without carotid disease (CAD). The study sample population was composed of 111
    patients treated at the Hospital Naval de Salvador; mean age 59.6 ± 13.9 years; all
    participants had whole blood DNA extraction, from that population, only 62 patients,
    mean age 57.85 ± 15.74 years, had carotid doppler performed. Then, PCR reaction
    followed by restriction endonucleases digestion (RFLP) were performed. Among patients
    who underwent carotid doppler sonography, 46.8% did not show CAD and 53.2%
    showed CAD. The frequency of the polymorphism was 81.98% for the wild homozygous
    (CC); 17.12% for heterozygote (CG); and 0.90% for mutant homozygote (GG).
    Individuals with the polymorphism exhibited significantly higher values for LDL-c (p =
    0.0201) and atherogenic coefficient (p <0.0001) with the same trend for non-HDL-c (p =
    0.0767) and for TG/HDL-c ratio (p = 0.0719). The frequency of the S447X polymorphism
    in the study population corroborated with that found in the literature, but its presence
    was not associated with the prevalence of CAD and with the comorbidities observed,
    such as diabetes mellitus, dyslipidemia and systemic arterial hypertension. AIP and
    other atherogenic indices can be used as discriminators of CAD.

15
  • RAIANA DOS ANJOS MORAES
  • ITACONIMIDATED DERIVATIVES INDICATE RELAXATION IN ARTERY MESSENTRICS AND NEGATIVE INOTROPISM IN RATS BY INHIBITION OF Ca2 + Influx savior
  • Advisor : DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • COMMITTEE MEMBERS :
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • DANIEL PEREIRA BEZERRA
  • FABIANO ELIAS XAVIER
  • Data: Aug 20, 2018


  • Show Abstract
  • Introduction: Hypertension is a risk factor for various cardiovascular and renal diseases, representing a major public health challenge. Thus, investigating new substances with antihypertensive potential becomes necessary. In this way, analogues were synthesized, N-4-methyl-phenyl-itaconimide (3), N-4-methoxy-phenyl-itaconimide (6), N-phenyl-itaconimide (9) and N-4-chloro-phenyl-itaconimide (15), from unprecedented alkaloid, phylantimide, available in Phyllanthus sellowianu. Objectives: To evaluate the effects of cyclic imides on cardiovascular system and to investigate possible mechanisms of action involved in observed responses. Methods: In vitro studies, Wistar rats were euthanized in CO2 chamber and superior mesenteric artery and atria were removed. In mesenteric arteries were performed the measurement of isometric tension, in atria were evaluated the heart rate and force of cardiac contraction, and in vivo studies was evaluated the actions of imide 6 on blood pressure and heart rate CEUA/UFBA (nº120/2017). Results: Cumulative administration of cyclic imides (3x10-8 to 3x10-4M) in pre-contracted rings with phenylephrine, 1μM, induced vasorelaxation that was independent of endothelium-derived relaxing factors. Additionally, imide 9 presented higher potency in relation to imide 3, 6 and 15 and not statistical difference was observed among the maximum efficacies of the imides. The cumulative administration of imides to basal tonus did not alter vascular intrinsic tone. In addition, imide 6 induced vasorelaxation in rings exposed to depolarizing tyrode solution with 60 mM KCl or 20 mM KCl similar to the control, suggesting the non-participation of K+ channels. Imide 6 was able to attenuate the influx of Ca2+ in a concentration-dependent manner. As well as, imide 6 attenuated the CaCl2-induced contraction in nominally calcium-free medium in the presence of cyclopiazonic acid (20 μM), phenylephrine (1 μM) and nifedipine (1 μM), indicating an attenuation of the influx of Ca2+ by receptor-operated channel (ROC) and store-operated channel (SOC). The presence of SKF 96365 (10-5M), SOC blocker, did not significantly alter the vasodilatory effect induced by imide 6. In addition, imida 6 induced negative inotropic effect without significant change in cardiac rhythmicity. In vivo studies, in non-anesthetized normotensive rats, imide 6 lowered blood pressure and induced bradycardia. Conclusion: These results suggest that imides have concentration-dependent vascular effects and the vasorelaxation seems to be independent of endothelium-derived relaxing factors. The vasodilatory effect induced by imide 6 may be due to a possible influence on the CaV and ROC channels. In addition, imide 6 is able to reduce force of cardiac contraction, blood pressure and promote bradycardia.

16
  • ALAN OLIVEIRA DUARTE
  • SOROLOGICAL PROFILE FOR ZIKA VIRUS INFECTION PARENTS AND NEWBORN IN ONE MATERNITY OF REFERENCE OF SALVADOR-BA
  • Advisor : FERNANDA WASHINGTON DE MENDONCA LIMA
  • COMMITTEE MEMBERS :
  • FERNANDA WASHINGTON DE MENDONCA LIMA
  • LUCIANA SANTOS CARDOSO
  • RICARDO RICCIO OLIVEIRA
  • Data: Aug 27, 2018


  • Show Abstract
  • In 2015, the first cases of zika virus (ZIKV) infection in Brazil were confirmed.
    After six months, cases of microcephaly associated with the outbreak occurred,
    which occurred concomitantly with outbreaks of dengue (DENV) and
    chikungunya (CHIKV). Studies are needed to understand the consequences for
    our population of this contact with the triple epidemic, especially in the pregnant
    population. Our objective was to investigate the serological profile for these
    arboviruses in puerperal women and newborn infants attended at a maternity
    hospital in Salvador, Bahia, Brazil in 2016. We performed a cross-sectional,
    seroepidemiological study that evaluated the presence of antibodies in mothers
    attended by the maternity ward who presented exanthematic disease in
    pregnancy and her neonates. ELISA diagnostic kits (Euroimmun™) were used
    to detect IgM and IgG against ZIKV, DENV and CHIKV, and a standardized
    anti-ZIKV-specific IgM capture ELISA was used. 101 mothers were included in
    the study. Regarding seroprevalence, for anti-ZIKV IgM, 07 mothers (6.9%) and
    04 neonates (4%) were reactive. For anti-CHIKV IgM, 23 mothers (22.8%) and
    04 neonates (4%) were reactive. For anti-DENV IgM, 12 mothers (11.9%)
    tested positive. Of the 101 mothers, 73 (72.3% of the total), 39 (38.6%) and 93
    (92.1%) were seropositive for IgG against ZIKV, DENV and CHIKV,
    respectively. Of the 102 neonates, 06 (5.9%) had microcephaly. We generated
    a new knowledge about the rates of congenital infection by ZIKV and CHIKV,
    and how these are expressed among mothers who presented exanthematous
    disease. Knowledge of seroprevalence for arboviruses is important to
    understand the extent of the arbovirus epidemic in Brazil, to evaluate groups
    susceptible to severe forms of disease and to stimulate prevention measures
    directed at these groups.

17
  • CATARINA MILENA MONTEIRO DA COSTA
  • INVESTIGATION OF THE REGENERATIVE THERAPEUTIC PROFILE OF THERAPY
    CELLULAR IN EXPERIMENTAL TRIGEMINAL NEURALGIA

  • Advisor : CRISTIANE FLORA VILLARREAL
  • COMMITTEE MEMBERS :
  • CRISTIANE FLORA VILLARREAL
  • DANIEL PEREIRA BEZERRA
  • PAULO JOSE LIMA JUIZ
  • Data: Aug 31, 2018


  • Show Abstract
  • INTRODUCTION: Among the various modalities of orofacial neuropathic pain, trigeminal neuralgia (NT) is the most common. It is associated with a variety of pathological conditions. The currently available treatments for trigeminal neuralgia, whether pharmacological or surgical, are not effective, highlighting the need to develop new therapeutic approaches for this syndrome. An emerging strategy is the use of stem cells. The therapeutic potential of these cells comes from their multiple sources of production, their mechanism of action and their regenerative potential. Stem cells have antinociceptive effect in trigeminal neuralgia, however the mechanisms involved in these effects are not yet well understood. The antinociceptive effect of stem cells in trigeminal neuralgia has been demonstrated, however, the mechanisms involved in these effects are not yet well understood. AIM: The purpose of this study was to evaluate the therapeutic potential of bone marrow mesenchymal stem cells (BMSC) and bone marrow mononuclear cells (CMMC) in the experimental model of trigeminal neuralgia, comparing their effects to those of pharmacological therapy and investigating their mechanisms of action. METHODS: BMSC were obtained from C57B16 mice and were characterized by flow cytometry and cell differentiation. Male C57Bl / 8 mice underwent surgery for induction of the neuropathy model by partial ligation of the infraorbital nerve and during 30 days were evaluated for body weight and thermal and mechanical nociceptive thresholds in the hargreaves and von Frey tests, respectively. Five days after induction of the model, the animals received single intravenous administration of CMMC (1x106), BMSC (1x106) or vehicle (100μ). The effects of cell therapy on the cytokine profile and structural alterations in the infraorbital nerve were investigated 10 and 30 days after the treatments, by ELISA and 30 (30 mg / kg), used as reference pharmacological treatment, by intraperitoneal route. days by optical and electron microscopy, respectively. RESULTS: BMSC and CMC transplantation completely reversed the thermal hyperalgesia (p <0.001) and mechanical allodynia (p <0.001) of neuropathic animals throughout the experimental period. As expected, carbamazepine induced short-term antinociceptive effect. The quantitative analysis of IL-1β, TNF-α, IL-1 cytokines in the infraorbital nerve of the mice showed that CMMO or CMsMO reduced IL-1β, TNF-α (p <0.001), and increased levels of IL-10 (p <0.001). Supporting the observed long-term antinociceptive effect, cellular therapy reduced structural alterations in the infraorbital nerve of neuropathic animals, with the reduction of axonal degeneration in myelinic and myelinic fibers and mitochondrial atypia. CONCLUSION: Treatment with BMSC and CMMC induced a consistent and long-lasting antinociceptive effect, associated with pro-and anti-inflammatory cytokine balance modulation, as well as the reduction of structural alterations in the peripheral nerve. These data show the potential of cellular therapy in the treatment of trigeminal neuralgia .

18
  • LILIANE BARRETO DA SILVA
  • Potential anti-hypertensive of carvacrol uncomplexed or complexed with β-cyclodextrin in spontaneously hypertensive animals.

  • Advisor : DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • COMMITTEE MEMBERS :
  • DARIZY FLAVIA SILVA AMORIM DE VASCONCELOS
  • DIOGO RODRIGO DE MAGALHÃES MOREIRA
  • RODRIGO MOLINI LEAO
  • Data: Sep 14, 2018


  • Show Abstract
  • Introduction: The phenolic monoterpene, carvacrol, has been used as a therapeutic agent in various cardiovascular diseases, but also the physical characteristics, such as liposolubility and volatility, limit sweetness in the compound. Thus, inclusion systems of drugs such as cyclodextrins appear to allow the development of a new strategy for the use of monoterpenes in therapeutic administrations. Objective: To evaluate the antihypertensive potential of carvacrol in the free or complex form of a β- cyclodextrin (β-CD) and whether the complexation is pharmacologically favorable for the cardiovascular actions induced by this monoterpene. Methods: In vivo studies, was evaluated the actions of CARV/β-CD and carvacrol, and daily dose at a dose of 50 mg / kg, on blood pressure and heart rate, after acute oral administration. The experiments were performed in spontaneously hypertensive rats (SHR) and normotensive controls rats (wistar). In the subchronic treatment, in 21 days, was investigated antihypertensive activity of both compounds. In vitro studies, the rats were euthanized in CO2 chamber and superior mesenteric artery, spleen and aorta were removed. In mesenteric arteries were performed the vascular reactivity, in spleen and aorta were evaluated the inflammatory and anti-inflammatory cytokines levels. Results: The acute oral administration in non-anesthetized normotensive rats and SHR rats, with carvacrol and CARV/β-CD, did not produce significant changes in blood pressure levels and heart rate. In the 21-day subchronic treatment, only CARV/β-CD induced a significant reduction in blood pressure levels in SHR rats. CARV/ β-CD was able to prevent the development of hypertension in SHR rats. Additionally, the vascular reactivity assays with SHR rats, presented higher potency in relation the -1 adrenergic agonist (phenylephrine), in relation to normotensive rats. Sensitivity to sodium nitroprusside did not change in all experimental groups.The levels of the pro-inflammatory cytokine IL-1β in the spleen were significantly reduced in the CARV/ β-CD treated SHR rats, and the levels of IL-10 in the aorta increased in these animals. Conclusion: CARV/β-CD was able to prevent the development of hypertension in SHR rats. In addition, it reduced levels of the pro- inflammatory cytokine- IL-1β, in the spleen, and increase the levels of the anti-inflammatory cytokine IL-10, in the aorta of CARV/β-CD treated SHR rats.

19
  • JONHY HERBERT GONÇALVES EVANGELISTA
  • SCREENING OF BINDERS
    FROM TRIPANOTIONA REDUTASE from
    Tripanosoma Cruzi

  • Advisor : MARCELO SANTOS CASTILHO
  • COMMITTEE MEMBERS :
  • MARCELO SANTOS CASTILHO
  • RAQUEL GUIMARÃES BENEVIDES
  • RICARDO DAVID COUTO
  • Data: Sep 27, 2018


  • Show Abstract
  • Chagas disease is a neglected tropical disease caused by the protozoan Trypanosoma cruzi that affects part of the world population, and is especially acute in Brazil, where it estimated that 1.9 million people are infected by T. cruzi and approximately 21.8 million are exposed to the risk in endemic areas of the country. Although it affects more than 1.8 million people in the Americas and puts at risk of contagion another 100 million, only two drugs are used to treat it (Benznidazole and Nifurtimox), which have serious adverse effects and high toxicity, making adherence to pharmacotherapy difficult. This scenario suggests the need to study new therapeutic targets, which are essential for the survival of the parasite, such as the oxidation pathway, in which the enzyme Trypanothione reductase (TcTR) plays a fundamental role. Although TcTR is a validated target, there are still no drugs that block this enzyme. This opens up a knowledge gap for the screening of inhibitors. Among the techniques of large-scale screening (HTS) the ThermoFAD technique can be highlighted. This methodology allows the investigation of thermal stability of Flavoproteins by monitoring the fluorescence signal of an intrinsic fluorophore (FAD) as a reflection of its unfolding. In order to identify ligands with minimum structural requirements for TcTR inhibition, the thermal detachment technique (ThermoFAD) was used for the screening of compounds. For this, we present the standardization of the chromatographic purification of TcTR and investigation by ThermoFAD of the conditions (TcTR concentration, pH, additives, etc.). Single dose screening allowed the identification of compound JU298, for which concentration-response assays were also conducted, confirming its stabilizing effect on TcTR. Such data therefore suggest that the JU298 compound as a potential inhibitor for the therapeutic target under study.

20
  • SANDRA CRISTINA HERNANDES
  • IDENTIFICATION OF VARIABLES TO BE INVESTIGATED AS
    PREDICTORS OF DRUG-RELATED PROBLEMS (PRM)
    TO RATIONALIZE THE CLINICAL PHARMACY IN A HOSPITAL
    Tertiary

  • Advisor : DENIS DE MELO SOARES
  • COMMITTEE MEMBERS :
  • DENIS DE MELO SOARES
  • PABLO DE MOURA SANTOS
  • RICARDO DAVID COUTO
  • Data: Dec 11, 2018


  • Show Abstract
  • Introduction: This study aims to identify risk factors for the occurrence of Drug Related Problems (DRP) in patients admitted to a tertiary hospital in the city of Salvador. DRP can cause preventable damage. Methodology: This study was an observational, cross-sectional study of a retrospective nature, carried out in a large-scale, highly complex hospital with 352 beds, data from the year 2016. The prescriptions were evaluated by identifying DRP by pharmacists clinic. Results: The main DRPs observed are described in the literature and could have been avoided emphasizing the importance of prevention measures. Discussion: The presence of the pharmacist in the review of medical prescriptions plays an essential role in the activities of hospital clinical pharmacists and can collaborate to improve the quality of drug use as well as patient safety. Conclusion: Knowing the risk factors for the occurrence of DRP is of paramount importance in establishing prevention tools and strategies.

21
  • KELVIS TRINDADE SANTOS
  • Evaluation of the antioxidant activity of Theobroma cacao L.
    beans husk extracts for cosmetic applicability

  • Advisor : EUDES DA SILVA VELOZO
  • COMMITTEE MEMBERS :
  • NEILA DE PAULA PEREIRA
  • RENATA BIEGELMEYER DA SILVA RAMBO
  • SERGIO EDUARDO SOARES
  • Data: Dec 17, 2018


  • Show Abstract
  • The Theobroma cacao L. beans husk is a waste in the industry known to
    contain a large amount of polyphenols and dietary fiber. The presence of
    polyphenolic antioxidants present in cocoa has caused an increasing interest in
    the use of products derived from cocoa in cosmetic preparations. This work
    aimed to obtain gels with antioxidant activity through the incorporation of
    extracts prepared from cocoa beans husks. The study addresses two distinct
    phases: the first one concerning the extraction of hydroethanolic and
    hydroglycolic extracts, fluid and dry, of the cocoa beans husk, which also
    includes the verification of physico-chemical parameters (pH, density and
    thermogravimetry), total phenolics and antioxidant activity (DPPH method). The
    second phase refers to the use of these extracts in gelled cosmetic formulations
    that were developed with different natural polymers (xanthan gum,
    carboxymethylcellulose and hydroxyethylcellulose), as well as physical and
    chemical parameters (pH and density) and stability (organoleptic
    characteristics, pH, density, texture and scattering in vitro) over time and under
    accelerated conditions. The antioxidant activity of the formulations was also
    evaluated. The extracts presented good results regarding their physicochemical
    profile, where they had values close to pH and density, besides being safe
    when applied in cold or hot formulations. It presented a high antioxidant
    potential in the extracts, highlighting the hydroethanolic, obtaining 74.72% SRL
    for the fluid and close to 90% SRL for the dry, besides an IC50 of 0.24 mg/ml for

    hydroethanolic and 1,15 mg/ml for the hydroglycolic. The formulations had their
    potential antioxidants proven, with a highlight for the gel with xanthan gum and
    hydroethanolic dry extract with value close to 80%. In addition, all the gels
    presented good performance of in vitro spreadability and texture, making them
    more stable when applying the extracts, preserving their values of spreadability,
    strength and adhesiveness. Based on the results obtained, cocoa beans husks
    can be used as a source of interesting raw material for cosmetology, especially
    in anti-aging dermotherapies.

2017
Dissertations
1
  • ALINE GRAZIELLE MAGALHÃES QUADROS MOREIRA
  • STRATEGIES FOR THE RECOMBINANT PRODUCTION OF LACASE ENZYME OF Cryptococcus neoformans
  • Advisor : MARCELO SANTOS CASTILHO
  • COMMITTEE MEMBERS :
  • MARCELO SANTOS CASTILHO
  • TANIA FRAGA BARROS
  • ACÁSSIA BENJAMIN LEAL PIRES
  • Data: Oct 2, 2017


  • Show Abstract
  • Cryptococcosis is an opportunistic infection caused by encapsulated fungi of the Cryptococcus neoformans complex distributed worldwide, mainly affecting immunodepressed patients. Antifungal agents available for treatment with intense adverse effects, and the difficulty for therapeutic adherence. In addition, the development of resources by strains of the fungus to drugs is a reality. This context points to the urgent need for new alternatives for the treatment of cryptococcosis. This scenario, a modulation of fungal virulence factors is a strategy that stands out as it differs in drug approaches, so please be more vulnerable to host defense. In this sense, a production of melanin that is a fundamental virulence factor for the development of the non-host microorganism and the laccase that catalyzes this reaction is a target for drug planning for the control of cryptococcosis. Thus, the objective of the present work was recombinantly produced the enzyme laccase of Cryptococcus neoformans (CnLAC1).In this perspective, CnLAC1 was cloned into the vector pET-28a(+) and subcloned the vector pET-32a(+) and expression assays performed to investigate the best soluble expression condition of the laccase. For different strains of E. coli and varying conditions such as temperature, incubation time and IPTG concentration were employed, all assays resulted in insoluble laccase. In view of this, structure of solubilization of inclusion bodies through the addition of detergents, variation of the urea concentration and variation of the pH of the buffer that is in the attempt to perform unsuccessful protein refolding. On the other hand, Arctic express cells were able to express soluble laccases, but have been co-purified with chaperone molecules. In view of this, a great challenge can be faced in the standardization of lactation purification to obtain a pure and active protein. Thus, the work described a variety of hierarchical strategies for the expression of the recombinant laccase enzyme, lacking current qualities of laccasekinetic and enzymatic characterization studies.

2
  • LENISA DANDARA DOS SANTOS
  • HUMAN HUMAN MPGES-1 EXPRESSION AND EVALUATION OF THE EFFECT OF ANTIPIRETIC SUBSTANCES ON THEIR GENE EXPRESSION.
  • Advisor : MARCELO SANTOS CASTILHO
  • COMMITTEE MEMBERS :
  • CARLOS EDUARDO SAMPAIO GUEDES
  • JOICE NEVES REIS PEDREIRA
  • MARCELO SANTOS CASTILHO
  • Data: Nov 23, 2017


  • Show Abstract
  • The use of anti-inflammatory drugs available in therapy is related to a number of adverse and/or side effects. As a strategy to develop safer drugs, selective inhibitors of prostaglandin E-synthase microsomal -1 (mPGES-1) have been developed. Through in vivo and in vitro phenotypic assays, an unprecedented series of antipyretic substances potentially inhibiting mPGES-1 was identified. However, it was not possible to determine the molecular mechanism of action through which these components exert an antipyretic effect. Therefore, this work has evaluated the activity of antipyretic substances probably inhibitory of mPGES-1 in in vitro assays. For this, mPGES-1 was expressed heterologously in E.coli and assays were performed on LPS-induced RAW 264.7 macrophages to evaluate the effect of antipyretic substances (10 μM) on mPGES-1 mRNA expression by RT-qPCR. Six substances were tested (compound 8; 9; 10; 15; 16 and 17) among which compounds 8; 9; 15 were able to reduce (p <0.05) the mRNA expression of the enzyme when purchased from the control (vehicle / LPS) group, whereas compounds 10; 16 and 17 had no effect. Based on these results, it can be concluded that the molecular mechanism of action by which compounds 8, 9 and 15 have caused antipyretic effect is, at least in part, via inhibition of mPGES-1 mRNA expression, while it is suggested that the compounds 10; 16 and 17 do not act in this way.

2016
Dissertations
1
  • BÁRBARA VELAME FERREIRA TEIXEIRA
  • EVALUATION OF POTENTIAL INHIBITORS OF PTERIDINE REDUTASE 1 (PTR1) AND REDUTASE-TIMIDYLATE SINTASE (DHFR-TS) DIHYDROFOLATE FROM Leishmania chagasi
  • Advisor : MARCELO SANTOS CASTILHO
  • COMMITTEE MEMBERS :
  • MARCELO SANTOS CASTILHO
  • RICARDO DAVID COUTO
  • GUSTAVO HENRIQUE GOULART TROSSINI
  • Data: Dec 16, 2016


  • Show Abstract
  • Among the parasitic tropical diseases, those caused by protozoa represent a challenge to public health. Leishmaniasis is a worldwide widely disease, affecting more than 90 countries. The main drugs used to treat this disease, as pentavalent antimonials, shows non-specific action and high toxicity. The Leishmania chagasi species is the responsible for visceral leishmaniasis, the most severe form of the disease, which has high incidence in Brazil. In trypanosomes, the folate biosynthesis pathway participates of the pteridine metabolism, role performed by the pteridine reductase enzymes 1 (PTR1) and thymidylate synthase reductase-dihydrofolate (DHFR-TS). The genus Leishmania parasites are auxotrophic for folates, therefore, molecules that act on PTR1 and DHFR-TS of Leishmania ssp. seem promising for the development of drugs against leishmaniasis. Therefore, this work aimed to determine the potency and mechanism of action, using enzymatic kinetics of pteridine derivatives, potential inhibitors of PTR1 and DHFR-TS. The compounds tested were active against LcPTR1 and LcDHFR-TS. The compound 5 have IC50 = 0.94 µM against LcPTR1 and had binding efficiency of 0.7 kcal.mol-1 . Atom – 1 which makes it a prototype compound for the development of a LcPTR1 and LcDHFR-T inhibitor in the future. Furthermore, inhibitors 1 and 5 showed competitive mechanism of action with the substrate and uncompetitive with NADPH. Therefore, this study allowed the kinetic characterization of a series of pteridine derivatives against LcPTR1 and LcDHFR-TS.

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