Different types of propolis are popular in various regions of Brazil, containing complex chemical components, mainly flavonoids and polyphenols, which vary according to geographical location, plant species, and the season in which they are produced. For example, green propolis is produced by Apis mellifera bees that use Baccharis dracunculifolia, a common species found in the Brazilian savannah. This study aims to evaluate Brazilian green propolis, both ''in natura'' and commercial, regarding its antioxidant, antimicrobial, cytotoxic, and neuroprotective potential, correlating these aspects with its chemical profile. To achieve this, propolis extracts ''in natura'' were used to prepare ethanolic, aqueous, and ethyl acetate extracts. The commercial aqueous and ethanolic extracts were obtained from Favo de Ouro Company, a laboratory partner. Phytochemical screening, a colorimetric and qualitative method, was used to characterize the extracts, assessing the presence of secondary metabolites. The crude extracts of green propolis and the aqueous and ethanolic solutions of commercial green propolis showed similarities in the presence of secondary metabolites. On the other hand, the aqueous and ethanolic extracts of green propolis ''in natura'' stood out due to the presence of saponins. Antimicrobial activity was evaluated by broth dilution. The crude extracts in ethanol and ethyl acetate proved effective against Staphylococcus aureus at concentrations of 125 and 250 μg/mL, respectively. The commercial aqueous solution was also effective against Escherichia coli. Cytotoxic and neuroprotective evaluation in PC12 cells for 48 hours showed that most extracts were not toxic, except for the commercial ethanolic solution, which exhibited toxicity. Regarding neuroprotective activity, the aqueous and ethyl acetate crude extracts, at concentrations of 11 and 5.5 μg/mL, respectively, showed promising results. Propolis analysis by GC-MS enabled the identification of a variety of compounds, such as terpenes, phenols, acids, aldehydes, and fatty acids, depending on the specific composition of the sample. This technique is essential for a detailed characterization of the chemical constituents present in propolis, providing valuable insights into its chemical composition.

Parkinson's disease (PD) is a public health problem that affects many individuals in the world, including Brazil. It is a neurodegenerative disease of slow progression that mainly affects the elderly population. Although scientific research has advanced in order to seek effective treatments for this pathology, there is no therapy capable of cure or inhibiting the progression of the disease. The scientific community has been prospecting neuroprotective compounds, which inhibit dysfunctions and death in cells involved in PD pathogenesis, among them, it is possible to cite the flavonoid rutin, which has had neuroprotective potential against glutamatergic excitotoxicity and cytotoxicity induced by 6-hydroxydopamin. However, the lack of information on the disease etiology difficult advances in the discovery of new drugs to treat PD. An important change that precedes the classic motor symptoms of PD is the disorder in gastrointestinal motility, associated with changes in the enteric nervous system, which also has unknown etiology. In vitro and in vivo study models have revealed that many cellular and molecular changes present in patients with the disease are related to aminochrome toxicity, a compound derived from dopamine and precursor of neuromelanine, present in remnant dopaminergic neurons.However, there is still no report that it is able to induce changes in the enteric function. The overall objective of this project was to evaluate the effect of rutin in Wistar rats with damage induced by aminochrome in terms of: 1- levels of dopaminergic neuron marker expression (tyrosine hydroxylase) in the ventral tegmental area (VTA) and substantia nigra parscompacta (SNpc) and 2- gastrointestinal traffic time. The animals were randomized in four groups: control group (CTR), aminochrome (AMI), rutin (RUT) and aminochrome + rutin (AMI+RUT). Rutin was administered by gavage, from the first to the twentieth first day after stereotaxis.The living weight of the animals was evaluated from the day of stereotaxic surgery until the time of perfusion. Intestinal transit time was evaluated by the carmine red test, aiming to analyze the functional changes induced by the treatments.As a result, rutin promoted a reduction in intestinal timely time compared to the control group. The aminochrome also reduced the expression of tyrosine hydroxylase (TH+) on the ipsilateral side of the SNpc,which was prevented by rutin treatment. However, no change in TH+ expression in VTA of animals treated with aminochrome and/or rutin was observed.Rutin promoted a reduction in the weight of treated animals, with a modulatory role in weight regulation. Thus, studies are necessary to evaluate the incited biochemical mechanisms for such modulation, which is important for further research using preclinical DP models

INTRODUCTION: Spinal cord injury is a serious damage to the central nervous system  associated with high rates of morbidity and mortality, physical dependence, and  financial burdens. Inflammation is the main characteristic of secondary injury that  provides a cascade of cellular and molecular events that increase the area of injury,  aggravating neurological deficits and tissue recovery. The literature presents different  models of studies in the central nervous system in which apigenin has demonstrated  beneficial effects. However, there is little research on the neuroprotective role of  apigenin in spinal cord injury, and in addition, the low solubility of this flavonoid in water  may be a limiting factor for its biological activities. Cyclodextrins are cyclic  oligosaccharides whose chemical structure allows the incorporation of lipophilic  substances in their cavity, giving these molecules greater stability and bioavailability.  OBJECTIVE: To evaluate the anti-inflammatory and neuroprotective effect of flavonoid  apigenin and conjugates of apigenin with β-cyclodextrins in an in vitro model of  neuroinflammation in spinal cord cells. MATERIALS AND METHODS: In this work β cyclodextrin (β-CD) and three other derivatives were used. The cytotoxicity of apigenin  (API) and conjugates of apigenin with β-cyclodextrins (API-cd’s) was evaluated by MTT  assay in PC-12 cells and spinal cord cells in a series of concentrations (0.1-100µM)  for 24h. After its characterization, the primary culture of the spinal cord was submitted to inflammatory damage by LPS, and subsequently treated with apigenin and the best  conjugates of apigenin. The investigation of changes in markers associated with  neuroinflammation was performed by immunofluorescence. The nitric oxide production  was analyzed with the supernatants of the experimental groups. The analysis of cell  morphology was performed by phase contrast microscopy, Rosenfeld staining and  immunofluorescence. Cell migration was evaluated by the wound healing assay and  immunofluorescence. RESULTS AND DISCUSSION: The API and API-cd’s showed  no toxicity in PC-12 cells at the concentrations tested. It was observed that three  conjugates increased the viability of PC-12 cells, mainly at the concentration of 10µm. The API and API-cd’s didn’t present cytotoxicity in primary culture of the spinal cord at  the concentrations tested. The primary culture of spinal cord of neonate rats presents  morphological diversity of astrocytes, microglia, and neurons, as well as cell growth  clusters. Spinal cord cells were subjected to inflammatory damage by LPS at five µg/ml  for 12 hours. The API and an API-cd 52 decreased the expression of IBA-1 and CD 68-positive cells after LPS-induced damage. Two API-cd’s significantly reduced nitric  oxide levels. Two API-cd’s significantly reduced the number of S100β-positive cells.  CONCLUSIONS: Apigenin and apigenin conjugates appear to protect spinal cord cells  in an in vitro model of neuroinflammation. 

To assess seed vigor, it is possible to perform accelerated aging (AA), which consists of submitting the seeds to high temperature and humidity. In addition, this test makes it possible to evaluate the behavior of seeds at a physiological, biochemical and molecular level. The objective of this study was to evaluate the behavior of seeds of cultivars BRS 188 Paraguaçu and BRS 149 Nordestina of Ricinus communis L. before and after being submitted to AA regarding the metabolomic, transcriptomic, enzymatic and physiological profile. The following analyzes were carried out: initial characterization of seeds, AA standardization, seed moisture contente, germination test, determination of electrical conductivity and pH exudate, tetrazolium test, lipid peroxidation (malonaldehyde levels - MDA), determination of the activities of antioxidant enzymes (superoxide dismutase – SOD, catalase – CAT, ascorbate peroxidase – APX, monodehydroascorbate reductase – MDHAR, dehydroascorbate reductase – DHAR, glutathione reductase – GR, glutathione peroxidase – GPX and glutathione S-transferase – GST). The evaluation of primary and secondary metabolites by metabolomics, and the production of transcripts through the RNA-seq test for the seeds of BRS 188 Paraguaçu. In the BRS 188 Paraguaçu seeds after the AA, there was a loss of vigor, increase in electrical conductivity, alteration in the production of primary (galactinol, myo-inossitol, melibiose, glyoxylic acid, sorbitol, inositol, xylose, sucrose and ribitol) and secondary metabolites of the terpene, phenolic and alkaloid classes, and greater gene expression, especially energy metabolismo. In response to AA, the increase in MDA concentration and APX and CAT activity was only significant in the whole seed, while SOD and DHAR were enzymes that showed a significant increase in activity in the whole seed and in the embryo. The change in GST and GPX activity was significant only in embryo samples. The cultivar BRS 149 Nordestina, showed after AA a significant reduction in viability, with membrane damage and increased lipid peroxidation. The increase in SOD activity, APX and DHAR were significant in whole seeds and the embryo, while CAT activity in the whole seed. MDHAR, GR and GPX, activity increased significantly only in the embryo. When analyzing the physiological, biochemical and molecular mechanisms of seeds after AE, that MDA, SOD and DHAR, and the increase of electrolytes in the exudate are biochemical biomarkers for the evaluation of seed quality, even as, the genes important for energy metabolism are suitable biomarkers for evaluating the quality of seeds of this species. 

Emerging infections caused by multidrug-resistant bacteria represent a major public health concern, as they pose a risk of nosocomial outbreaks, especially in immunocompromised patients. The non-diphtheritic bacteria of the genus Corynebacterium are Gram-positive microorganisms that may be present in the normal human microbiota, but isolates from these species showing the phenotype of multidrug resistance have been recently identified in several cases of infections worldwide. Therefore, it urges the understanding of the genetic factors involved in virulence and antimicrobial resistance in these species. This thesis is based on the hypothesis that the comparative genomic analysis of several clinical isolates of non-diphtheric Corynebacterium spp. will allow for a better understanding of the genetic mechanisms involved in the transition of these species to the pathogenic phenotype in humans, in addition to an understanding about the mechanisms involved with the rapid acquisition of multiple resistance to antimicrobials. For this, we performed the whole genomic sequencing of 11 clinically relevant isolates, 3 of the Corynebacterium striatum species isolated in Brazil, and 8 of the Corynebacterium amycolatum species, of which 6 were obtained in Spain and 2 in Tunisia. We obtained 24 genomes from C. striatum and another 18 from C. amycolatum deposited at the NCBI to integrate our set of analyses. We developed an analysis pipeline that starts with phylogenomic identification by ANIb in JSpecies, to ensure the analysis of genomes with accurate taxonomic identification, pan-genomic analysis with BPGA, a pipeline tool that uses USEARCH to perform gene clusters, in addition to the COG functional annotation of the pan-genome. We applied our genomes in PATRIC to predict antimicrobial resistance (AMR) genes, from the CARD and NDARO databases, in addition, we performed predictions on other web platforms such as VFanalyzer for virulence factors and IslandViewer4 for genomic islands. The pan-genomes of both species have open status, C. striatum presented α = 0.852803 with a pan-genome containing 3816 gene families (27 genomes), and C. amycolatum α = 0.854905 with its pan-genome containing 3280 families of genes (26 genomes). We identified 15 AMR genes and 32 virulence factors in the analyzed set of genomes of C. striatum, whereas in C. amycolatum we identified 9 AMR genes and a variety of 47 virulence factors. Analyzing the predicted genomic islands, we identified AMR genes and virulence factors contained in this genomic context in both species. These analyzes demonstrated the relevance of acquiring AMR genes and virulence factors through horizontal transfer, and data on pan-genome status support this feature in these species.

Ricinus communis L., known as castor bean is an oilseed species, grown worldwide in the tropical area e has great socioeconomics value due to several applications of castor bean oil. Temperature is a critical environment factor in vegetal development, variations in this parameter can lead to significant physiological, biochemical and molecular alterations. In this context, the temperature effect in antioxidant activity and physiological response of imbibed seeds and seedlings of Ricinus communis L. Therefore, the activity of antioxidant enzymes (glutathione reductase – GR, glutathione-s-transferase-GST, dehydroascorbate reductase-DHAR, superoxide dismutase-SOD, catalase-CAT and ascorbate peroxidase) in seeds and seedlings of R. communis under different temperatures (25, 30 e 35°C) was determinated during germination and seedlings establishment. The activity of antioxidant enzymes decresead at 25°C condition and increased at 35°C. Although the 35°C condition accelerates germination, it can be considered a thermal stress condition and increased the number of deteriorated and deformed seedlings. Thus, the 30°C condition can be considered the best condition to germinate and grow Ricinus communis L. whereas the 25°C does not causes oxidative stress to seeds and seelings, it slows down the germination and growth. The antioxidant enzymes activity varies according to botanical part and development stage of vegetal organism. The thermotolerance of R. communis is associatated to a strong antioxidant complex, where superoxide dismutase was identified as a potential biomarker to heat stress in seeds and CAT for aerial part of Ricinus communis L. seedlings.

Rutin is one of the most common used polyphenolic compounds described in the literature, with great scientific prominence due to its interesting pharmacological activities for of human health, including its neuroprotective potential. In the context of neurodegenerative diseases, glutamatergic excitotoxicity has been identified as an important mechanism of neuronal death associated with progression of these diseases. Therefore, this study aimed to investigate the rutin neuroprotective activity for neural cells under cytotoxic glutamate action. For this, cells of the PC12 strain were subjected to different treatments: direct treatment with rutin (0.5 - 1 μM) and /or glutamate (10 - 60 mM); indirect treatment using conditioned medium obtained from brain organotypic culture of Wistar rat (p7-p9) treated with rutin (0.5 μM) and glutamate (60 mM). In addition, we also treated primary culture of mesencephalic neurons/glial cells with 0.5 μM rutin. After 24 hours of direct or indirect treatment of PC12 cells, we performed cell viability analyzes from the Trypan Blue and Propidium Iodide exclusion test. In addition, we performed morphological analyzes after Rosenfeld staining for PC12 cells and for primary culture of mesencephalic neurons/glial cells. The results obtained with the cell viability test demonstrated that rutin was not toxic to PC12 cells. Glutamate, in turn, induced a toxic effect in PC12 cells in a concentration-dependent manner, and the concentration at 60 mM was able to cause almost 100% of death to the cells in culture. Regarding the direct neuroprotection assays, it was shown that, compared to the control group that presented 2% death, rutin was not able to protect PC12 cells against glutamate toxicity at high concentrations (30 and 60 mM) inducing about 20% and 100% death respectively. On the other hand, conditioned media from brain organotypic culture treated with rutin- (0.5 μM) + glutamate (60 mM) induced low toxicity in PC12 cultures, more specifically only 8% of cells died with treatment, whereas direct treatment presented 100% dead cells. Regarding morphological changes, indirect treatment with rutin induced a characteristic change in neuronal differentiation for approximately 26% of cells in culture, unlike direct treatment which induced 2.5%. These morphological changes related to the increase of branch points and the increase of neurite length were observed in PC12 cells and in primary culture of mesencephalic neurons/glial cells. These results indicate that the neuroprotective and morphogenic effects of rutin may be associated with modulation of glutamate metabolism by astrocytes and/ or release of soluble neuroprotective and inductors of neural differentiation factors.

INTRODUCTION: Lepidium meyenii is a plant that has several medicinal properties, however, the correlation between the metabolites and the biological activities tested in the plant is limited. OBJECTIVE: To characterize the metabolomic profile as well as to evaluate the antioxidant, antimicrobial and cytotoxic activities of extracts obtained from the dehydrated plant and its commercial products and to correlate the metabolomic profile with biological activities. MATERIALS AND METHODS: The extracts were obtained from the root of the plant and its commercial products by maceration in different organic solvents. The metabolomic profile was evaluated by mass performance coupled high chromatography (HPLC-MS) and gas chromatography coupled to mass spectrometry (GC-MS). The antioxidant activity was determined by the free radical sequestration method 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total phenols will be quantified by the Folin-Ciocalteu method. Antimicrobial activity was evaluated against bacteria and yeast by broth microdilution method, cytotoxicity was evaluated against C6 cell line and astrocytes. RESULTS AND DISCUSSION: The antioxidant activity (IC50) ranged from 64.97 μg∙mL-1 for the ethyl acetate extract from the root of the dehydrated plant to 935.61 μg∙mL-1 for the hexane extract of the plant commercial product. The total phenol values ranged from 6.83 mg∙GAE∙g-1 for the ethanol extract at 49.83 mg∙GAE∙g-1 for the ethyl acetate extract, both extracts are form the commercial products. The ethanol extracts and ethyl acetate were the most active and had the highest antioxidant potential, in addition they also showed antimicrobial activity against M. luteus and B. cereus and antitumor activity against C6 cells, with no cytotoxicity to astrocytes. Seventy-six metabolites were identified in the extracts and among these, terpenes were the main metabolites responsible for antioxidant and cytotoxic activities and fatty acids for antibacterial activity. CONCLUSION: The metabolomic approach through the several analytical techniques allowed to correlate the obtained results being possible to identify the main bioactive compounds of L. meyenii related to the antioxidant, antimicrobial and antitumor activities of the plant. 

Introduction: Diabetes Mellitus (DM) is a chronic disease developed by defects in insulin signaling, secretion or production. Prediabetes is a asymptomatic state in which subjects fasting glucose is between 100-120mg/dl and glycated hemoglobin is at the range of 5,7-6,5%. Defined before as a latent state it is now known that is a risk factor for nephro and neuropathy development and also increase in β-cell destruction. In this context, becomes necessary the identification of biological markers that presents a disrupted expression due to the pathological state thus contributing to an early diagnostic. miRNAs are non-coding RNA considered important in the pos-translation regulation of genes. They are pointed out as potential biomarkers, since their expression level becomes altered due to the development or aggravation of many diseases such as diabetes, cancer and schizophrenia. The present study aimed to identify an expression microRNA (miRNA) profile capable of confer early prediabetes diagnostic. Publicly available transcriptome was analyzed through bioinformatic tools. Results: After screening conducted at NCBI GEO datasets, one transcriptome matched our inclusion criteria. From which thirty-three miRNA were significatively upregulated in prediabetic subjects versus control (FC>1,5 p-valor≤ 0,05) and two were upregulated in the diabetix cersus control group (FC>1,5 p-valor≤ 0,05) A miRNA-mRNA network was created using the upregulated miRNA and its regulated targets in the validation transcriptome using the Cytoscaped software. Five miRNAs presented a greater centrality score from the miRNA-mRNA network. In which two (miR-93 and Let-7a) had better values for sensibility and specificity (AUC =0,829 e 0,824 respectively). Conclusion: The five miRNA identified are involved in several intracellular pathways related to insulin resistance and destruction of β-pancreatic cells and are differentially modulated in the transcriptome analyzed in this study, which can be identified as promising exploratory biomarkers for the early diagnosis of DM2.

Eating habits with high fat levels and sedentary lifestyle are triggers factors for obesity, type 2 diabetes (DM2), and heart failure. Mesenchymal stem cells (MSC) arise as a therapeutic tool capable of improving cardiovascular disease, but also of other degenerative conditions, due to its regenerative effect induced by its paracrine action. Based on this, the factors secreted in the conditioned medium (CM) of this cells are also investigated in tissue regeneration. Therefore, this study aimed to evaluate the therapeutic potential of MSC and its CM in the treatment of cardiac complications in an experimental model of obesity and diabetes induced by high fat diet (HFD). Cardiac, murinometric and biochemistry evaluations were done at different periods such as pre-indcution, pos-induction and pos-treatment. After 36 weeks of HFD administration this diet were replaced by the standard diet and then mice were treated with MSC, MC and DMEM (vehicle). Cardiac function was assessed by electrocardiography, echocardiography, and treadmill test. Metabolic dysfunction was assessed through measurements of body weight and biochemical parameters. Molecular mechanisms of action of the treatment were investigated using qRT-PCR in cardiac tissue. The presence of cardiac fibrosis were evaluated by histopathological analysis. The characterization of MC by protein array showed the presence of 19 proteins including cytokines and growth factors. Mice fed HFD developed cardiac arrhythmias, expression alterations of cardiac genes and myocardial fibrosis, reflecting in the reduction of physical activity due to obesity and DM2. Administration of MSC or CM improved the severity arrhythmias and recovery exercise capacity. The first part of this study the improvement of cardiac function was in agreement with the expression normalization of GATA4, conexin 43 e COL1A1  genes in HFD mice treated with MSC ou MC. By other side the gene expression of troponin I, adiponectin, TGFβ, PPARγ, IGF-1, SOCS3, MMP9 and TIMP1 were normalized only after MSC treatment, but not with CM. Moreover, MSC or CM administration reduced the intensity of cardiac fibrosis. In order to elucidate the significant improvement achieved by MSC, in the second stage of this study, the investigation on the molecular mechanisms involved in this therapeutic effect. The expression of genes related to tissue remodeling (SPARC and CTGF), apoptosis (SMAD7 and PDCD4), inflammation (CCl2 and CHI3I3), as well as cell survival and proliferation (PTEN and STAT3) by qRT-PCR were investigated. High levels of gene expression of SMAD7, PDCD4, SPARC, CTGF, CCL2, STAT3 and PTEN were detected only in the MSC-treated group, whereas in CHI3I3 and NPPA gene expression was reduced in both MSC-treated and DMEM-treated mice. The results suggest that MSC and MC have a therapeutic effect on cardiac alterations due to obesity and T2DM, being the factors related to the development of fibrosis, apoptosis and cell survival modulated, demonstrating the relevance of the factors secreted by MSC reinforcing their action paracrine.

INTRODUCTION: Secondary treatments mechanism, such as the use of antioxidants and interferon (IFN) type 1, for the pathology associated with Human T-cell lymphotropic Virus type 1 (HTLV-1), HTLV-1 Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP), is not elucidated, but, it is based on protection against Reactive Oxygen Species (ROS). The human Superoxide Dismutase 1 (SOD1) enzyme catalyzes the dismutation of superoxide radicals and it is important for the HTLV-1 survival in the intracellular environment. Inhibitory mechanisms of this enzyme, using diethyldithiocarbamate (DETC) for example, could increase the intracellular superoxide concentration, triggering oxidative stress and inducing cell death trough apoptosis. Thus, the regulation of superoxide in infected lymphocytes through the SOD1 antioxidant activity could be a therapeutic target for Adult T-cell leukemia/lymphoma (ATLL), another pathology also associated with HTLV-1. OBJECTIVE: To investigating the antioxidant SOD1 enzyme as a potential therapeutic target for ATLL; and its inhibitor, DETC, as a promising pro-oxidant therapy for this pathology. MATERIAL AND METHODS: The Gene Expression Omnibus (GEO) database was used to the search for gene expression datasets associated with ATLL clinical forms (Indolent, chronic, lymphomatous, and acute). The meta-analysis of these data was performed using the Ingenuity Pathway Analysis (IPA) software. SOD1 plasmatic concentrations in symptomatic infected individuals (ATLL and HAM/TSP), asymptomatic carriers, and uninfected healthy subjects were evaluated by enzyme-linked immunosorbent assay ELISA Cu/Zn-SOD. MT2 and MT4 T lymphocytes were used in the in vitro assays to evaluate the basal cytoplasmatic concentration of SOD1 by Cu/Zn-SOD ELISA; and also, for evaluation of SOD1 inhibition using DETC. The DETC apoptotic capacity and the validation of its action using the antioxidant N-acetylcysteine (NAC) were evaluated by flow cytometry and quantified by staining the cells with Hoechst 33432. RESULTS AND DISCUSSION: In the chronic and acute clinical forms of ATLL, we have observed that the NFE2L2 gene, related to the antioxidant response regulation, is being differentially expressed, under positive regulation, and that the classical lymphocyte proliferation pathway is activated.
SOD1 plasmatic quantifications in healthy donors (28.3, 15.6-37.75 ng/mL, n = 20), asymptomatic carriers (193.6, 170.2-327.4 ng/mL, n = 19), HAM/TSP patients (38.1, 10.6-59.25 ng/mL, n = 14) and ATLL patients (270.6, 141.8-456.9 ng/mL, n = 25) demonstrated that SOD1 concentration is significantly lower (p <0.0001) in HAM/TSP individuals, compared to asymptomatic carriers and ATLL patients, which may explain the therapeutic success of antioxidants, such as vitamin C, used in this clinical form; SOD1 concentration is also significantly higher (p <0.0001) in asymptomatic carriers compared to HAM/TSP and healthy donors, which could suggest SOD1 as a prognostic marker for ATLL; and significantly higher (p <0.0001) in ATLL patients, when compared to HAM/TSP and healthy donor patients, which may represent a promising therapeutic target for the treatment of this pathology. It was also possible to demonstrate a gradual decrease of cell viability, of MT2 and MT4 lymphocytes, with the increase of DETC concentration, and a greater sensitivity, to this drug, of MT4 cells (IC50 = 11.92μM) when compared to MT2 cells (IC50 = 26.45μM). We also determined the maximum of positive response to DETC treatment that was superior in MT2 lymphocytes (50μM) compared to MT4 lymphocytes (10μM). The reversion of DETC action in the two type of cells using the antioxidant NAC, for validation of SOD1 inhibition by DETC, was more prominent in MT2 (Up to 500μM DETC) than in MT4 lymphocytes (Up to 10μM DETC). These results are corroborated by the fact that MT2 lymphocytes have higher SOD1 basal cytoplasmatic concentration (6.73, 4.47–10.43 ng/mL) than MT4 lymphocytes (3.15, 2.78–3.5 ng/mL). CONCLUSION: SOD1 could be suggested as a potential therapeutic target for ATLL, as well as the use of DETC as a promising pro-oxidant therapy in this pathology.

Medicinal plants have always been used for the treatment of diseases. In order to evaluate the metabolic profile and antioxidant, antimicrobial and cytotoxic properties of plants used by traditional communities in the Northeast region of Brazil, five medicinal plants (Abarema cochliacarpos, Euphorbia tirucalli, Alpinia zerumbet, Miconia albicans and Protium heptaphyllum) were selected based in the literature review and the use of these plants by these communities, as well as their therapeutic indications. Leaves, stems and bark of the plants were collected at the Sapiranga Reserve, Mata de São João - BA, dried at room temperature and ground. The extracts were obtained by maceration in ethanol. The antioxidant activity was determined by the elimination test of the 2,2-diphenyl-1-picryl-hydrazyl radical. Total phenolics were quantified by the Folin-Ciocalteau method. The antimicrobial activity was evaluated as minimum inhibitory concentration (MIC) using the microdilution assay in 96-well plates against Gram-positive Bacillus subtilis, Staphylococcus aureus, Streptococcus mutans and Micrococcus luteus and Gram-negative Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium. Cytotoxic activity was performed against nauplius II of Artemia salina. The metabolomic profile was performed using LC / MS. As for the antioxidant activity, the IC50 for A. zerumbet was 15.87 μg.mL-1 whereas for extracts of P. heptaphyllum it varied from 4.67 μg.mL-1 (bark) to 22.16 μg.mL-1 (leaves). For A. cochliacarpos the values ranged from 3.81 μg.mL-1 (stem) to 5.58 μg.mL-1(leaves). For M. albicans it varied from 4.63 μg.mL-1 (leaves) at 5.05 μg.mL-1 (stem). E. tirucalli presented the lowest value of 317.2 μg. mL-1. The highest values of total phenolics were evidenced in the extracts of A. cochliacarpos, ranging from 70.11 mgEGA.g-1 (leaves) to 100.22. mgEGA.g-1 (bark). The lowest value was obtained from E. tirucalli extract (9.65 μg.mL-1). For A. zerumbet the found value was 22.46 mgEGA.g-1. For P. heptaphyllum the values ranged from 24.66 mgEGA.g-1 (leaf) to 58.22 mgEGA.g-1 (bark). The total phenolic content of the extracts of Miconia albicans ranged from 49.70 to 52.71 mgEGA.g-1. The extract of M. albicans (stem) showed antimicrobial activity against Pa, Sa, Bs and Bc. Alpinia extract inhibited bacterial growth at concentrations of 500 μg. mL-1 for Bc, 125 μg. mL-1 for Sa and 500 μg. mL-1 for Ec. The antibacterial activity of the extracts of P. heptaphyllum was high with MIC of 15.62 μg.mL for Ml. The M. albicans extract demonstrated conspicuous antimicrobial activity against gram-positive bacteria. There was also activity against gram-negative Pa bacteria. All species exhibited lethality rate of 100% Artemia salina at the concentration of 100 μg. mL-1 extract. The metabolic analysis of extracts by HPLC/MS allowed the quantification of several secondary metabolites, such as flavonoids, phenol compounds, phenylpropanoids, terpenes and alkaloids, among others.

The HIV was discovered in 1983 and has killed more than 35 million people worldwide while currently 36.9 million are carriers of this virus. HIV-1 is the most widely distributed strain in the world and its high mutation rate is one of the most striking features, resulting in the emergence of strains with resistance to available antiretroviral drugs, which makes the development of new drugs vital. Fusion inhibitors are third-line antiretrovirals that prevent viral entry into host cells and Enfuvirtide is currently the only marketed antiretroviral of this class. Although there are tools that determines the resistance of HIV-1 strains to most drugs, there are still no specific software for fusion inhibitors. The aim of this work was the development of a free access tool to identify mutations associated with resistance of HIV-1 virus genotypes to antiretroviral fusion inhibitor drugs. Secondarily, this work evaluated the prevalence of Enfuvirtide resistance in Brazilian and world populations and compared the level of resistance between different viral genotypes. The HIVfird was developed using the PHP programming language and uses Kalign to perform DNA sequences alignments. As a reference for alignment with user-inserted sequences, the tool utilizes 30 DNA bases obtained from the HIV-1 HXB2 gp41protein (amino acids 36 to 45). To assess the level of resistance to Enfuvirtide in the populations studied, all HIV-1 sequences that possessed the genomic region associated with Enfuvirtide resistance were retrieved from the Los Alamos National Laboratory database. The HIVfird software is hosted at https://www.hivfird.ics.ufba.br/, fully functional and available for public use. Twenty-five amino acid substitutions that alone have caused some level of drug resistance and 15 combinations of 2 distinct substitutions associated with decreased susceptibility to Enfuvirtide have been identified in the fragment between amino acids 36 to 45 of the gp41 protein sequence. Positions 44, 43, 36 and 38 were the ones with the highest frequencies of variations associated with drug resistance. From this new tool, we characterize DNA sequences present in the Los Alamos database, where such substitutions were found in 3.16% in the world population sequences and 4.67% in Brazilian population sequences. The prevalence of resistance to Enfuvirtide was identified for the different subtypes of HIV and was found that the rate was significantly higher for subtype B compared to subtype C (p < 0,0001) and may be due to economical and geographical factors. Finally, it is expected that HIVfird will be added to a list of antiretrovirals analysis tools and assist health professionals in planning and monitoring Enfuvirtide use as well as population-based analyzes of drug resistance by the scientific community.

INTRODUCTION: The Human Immunodeficiency Virus (HIV) is the etiological agent of Acquired Immunodeficiency Syndrome (AIDS), a disease responsible for the deaths of 39 million people worldwide and still without a vaccine and without a cure. The virus has two types, 1 and 2, type 1 is divided into four groups M, N, O and P and group M is subdivided into 9 subtypes and recombinant forms. Subtype C accounts for more than 50% of HIV-1 infections worldwide, being first isolated in Ethiopia in the 1980s and found in populous countries of Africa, India, China, Australia, Europe and South America. In the southern region of Brazil, it is the predominant HIV-1 form and its circulation is also increasing in other regions of the country. Phylogenetic studies reconstructed the history of subtype C, evidencing its insertion in Brazil from a monophyletic origin in this region, relating to the lineages found in the countries of the central-east region of the African continent. In the Northeast, subtype C has been detected with a prevalence of 4-5% of HIV infections. OBJECTIVES: To investigate the origin of subtype C circulating in Northeast region and to evaluate its migratory profile in the Brazilian territory. MATERIALS AND METHODS: 48 pol gene sequences from Brazil and 36 from worldwide, isolated between 1998 and 2015, were collected from public databases. Sequences were aligned using MAFFT and Bioedit. Samples subtype was confirmed by phylogenetic analysis, using the Maximum Likelihood (ML) method, implemented in IqTree. The phylodynamic analysis was based on the Bayesian method using the BEAST software tools version 1.8.4. RESULTS: The Bayesian analysis indicates the occurrence of five introductions of subtype C in Brazil. Fourty four (91.7%) Brazilian subtype C sequences grouped into a monophyletic clade composed exclusively of C sequences from Brazil. This group would have originated from countries in East Africa like Burundi, Ethiopia and Kenia around 1983 (1975 to 1992). The other four subtype C introductions in Brasil date from 1985-1990. Three of them were are represented by sequences from Southeast of Brazil and would have emerged from South Africa, Tanzania and Zambia. The fifth subtype C insertion into Brazil is represented by a sequence from Roraima and would have originated from Tanzania, most likely in 1987. Multiple introductions of the subtype C main lineage occurred between 1986 and 2005 in Northeast region and would have originated from the other four Brazilian regions. CONCLUSIONS: The subtype C epidemic in Brazil results from multiple introductions of this variant from the countries of East and Southern Africa. Most of the viruses of this subtype circulating in the country, are derived from a single predominant lineage. For the first time, an independent introduction of subtype C in Northern Brazil was identified. An ongoing migration of the virus towards the North of the country is observed. The results show a wide diffusion of subtype C in the five Brazilian regions and emphasizes the role of Goias state in the dissemination of the virus to northern regions.