Melatonin is a hormone provided by the mother to the fetus during development, acting as an antioxidant in crucial cellular processes of the nervous system. The use of Valproic Acid (VPA) during pregnancy serves as a model for studying Autism Spectrum Disorder (ASD). We assessed whether the absence of maternal melatonin during intrauterine development results in damages similar to autism in offspring, as observed in animals exposed to VPA. Wistar rats were divided into five groups: pinealectomized with and without melatonin supplementation, a surgical procedure without pineal gland removal (SHAM), VPA injection on the 13th day of gestation, and saline injection as a VPA control. Two days after birth and at the fourth week of life, the offspring were sacrificed, and the neonatal brain and young rat cerebellum were collected and analyzed for GSH content, H₂O₂ production, lipid peroxidation, and Na,K-ATPase activity and its isoforms. Our neonatal results showed gender differences in oxidative stress levels, with males exhibiting higher levels than females. In the cerebellum, we observed similar effects in the offspring of pinealectomized rats and those exposed to VPA, including increased oxidative stress, lipid peroxidation, and decreased GSH, as well as deficits in Na,K-ATPase activity. Exogenous melatonin supplementation restored normal levels of oxidative parameters and the total activity and isoform α1-Na,K-ATPase in the cerebellum. In summary, this study shows that oxidative stress may play a crucial role in the etiology of ASD and underscores the importance of maternal melatonin in preventing neurodevelopmental disorders in offspring. Additionally, it points to possible gender differences that will be investigated in future studies.

Obesity is a chronic disease characterized by excessive accumulation of body fat, which can generate metabolic disorder in all systems, including the nervous system, producing a process of inflammation and mitochondrial dysfunction, mainly by excessive production of Reactive Oxygen Species (ROS). These disorders generate neuronal death and behavioral changes. Atorvastatin is a drug frequently used for hypercholesterolemia, almost always associated with obesity. The objective of this project was to evaluate behavioral and biochemical parameters in the hippocampus and cortex of male wistar rats fed a cafeteria-type diet and treated with atorvastatin. Male Wistar rats were divided into four groups: commercial diet - saline (CoSal), commercial diet - atorvastatin (CoArt), cafeteria diet - saline (CafSal), and cafeteria diet - atorvastatin (CafArt). Atorvastatin was administered by gavage at a dose of 10 mg/kg/day and saline at a dose of 0.9%, and the cafeteria groups received water with 20% sucrose. The animals were weighed daily and underwent behavioral anxiety testing on day 23 and memory tests on day 24. On the 25th day, the animals were euthanized, blood was collected for cholesterol quantification by colorimetric method, and the body was dissected for removal and weighing of retroperitoneal and epididymal fat. In addition, the cortex and hippocampus were dissected and stored in -80ºC freezer for further quantification of caspases and AIF by Western Blotting and for quantification of lipoperoxidation by TBARS method. The circulating cholesterol was affected by diet and normalized by atorvastatin. Regarding weight, there was no change in total weight, but the weighing of fat showed that there was accumulation of adipose tissue in the cafeteria animals. The cafeteria diet caused hypomobility and spatial memory damage in the animals, and atorvastatin inhibited these effects. The diet did not cause problems in de novo memory or anxiety, but the use of atorvastatin without the diet caused problems in these parameters. The expression of apoptotic proteins (AIF and caspases 8 and 9) was increased by the cafeteria diet and returned to control with the use of atorvastatin.

few decades ago we have been going through a dietary change, with increased consumption of high-calorie foods, together with a decrease in physical activity, contributing to the increase in adiposity in the world population. Patients with high adiposity may develop chronic diseases, such as type 2 diabetes mellitus, hypercholesterolemia and cardiovascular disease (CVD). In studies, it has been shown that autonomic imbalance plays an important role in the pathogenesis of several CVDs, increasing the risk of cardiac death. Thus, the aim of the present study was to evaluate the temporal effect of a diet with high levels of carbohydrates and lipids on the cardiovascular autonomic modulation in rats through the analysis of the electrocardiogram (ECG) record by linear and non-linear methods. Wistar rats were fed for 24 days with ration supplemented with items of high caloric density and water added with sucrose (20%) or with commercial ration and water ad libitum. ECG electrodes were made and implanted in both groups of animals. Before the surgical procedure, the rats received a mixture of ketamine and xylazine, veterinary pentabiotic and meloxicam (doses adjusted with body weight). On days 6, 12, 18 and 24 of the diet, the ECG of the rats was recorded and at the end of the experiment linear analyzes were performed in the time domain [mean and standard deviation (SDNN) and the square root of the mean of the differences between consecutive RR intervals squared (RMSSD)] and in the frequency domain [low (LF) and high (HF) frequency spectral bands and the LF/HF ratio] and the analysis by non-linear methods, symbolic dynamics [variation patterns of families (0V, 1V, 2UV and 2LV)] and multiscale entropy (scales from 1 to 25 and scale groupings 1 to 5 and 6 to 25). Heart rate (HR), cardiac interval (CI) and body weight gain were also evaluated. Normalities were tested using the Shapiro-Wilk test and significant differences were analyzed using the two-way anova test, except for the analysis of multiscale entropy (EMS) and body weight, which was evaluated using Student's t test. Data were expressed as mean ± SEM (P<0.05). The present experiment was approved by the Ethics Committee for the Use of Animals of the UFSJ (protocol 32/2017). In rats fed the control diet, it was observed that its development induced a decrease in sympathetic activity on days 12 and 18 of the diet, evaluated by the 0V family of symbolic dynamics, and on day 24 in the 1V family of symbolic dynamics. The HR of the animals fed the control diet, during their development, had an increase in CI/decrease in HR. Animals fed the cafeteria diet showed an increase in HR/decrease in CI. An increase in the LF band (%) and in the LF/HF ratio from the 12th day on the diet was found in animals fed on the cafeteria diet, as well as in the 0V family of the symbolic dynamics and a reduction in the 2UV family in this period, in relation to the control rats. These data suggest an increase in sympathetic nervous system activity (HR, IC, LF, LF/HF, 0V) in animals treated with the cafeteria diet and a decrease in parasympathetic activity (2UV). The multiscale entropy was increased only on the 12th day of the diet in scales from 3 to 25. It is possible that the rats fed the cafeteria diet had a higher entropy value than the rats fed the control diet, with lower physiological complexity. It is possible to conclude that during the development of the rat there is a decrease in sympathetic activity and an increase in parasympathetic activity for the heart. The cafeteria diet offered to the rats blocks this effect from the 12th day of offering the diet, as a result, the sympathetic activity to the heart remains increased in these animals. Thus, the cafeteria diet offered to the rats for a short period of time is enough to alter cardiac sympathovagal balance.

Disturbances in the skin microbiota are related to infections and inflammation that cause diseases such a acne and others. Alternatives for treatment are probiotic strains. Probiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. The application of lysed and inactivated in the cosmetic industry is a trend. As long as the beneficial effects are maintained, they have advantages such as transportation, shelf life, risk of translocation and consumer infection. The aim of this study was to evaluate the antimicrobial activity of cell-free, lysed and inactivated supernatant, viability, sublethal damage, influence of the culture medium on different methods, optimize biomass production and microencapsulation of lactic acid bacteria (LAB) isolates. Cell-free supernatants from the fifteen LAB strains did not inhibit the growth of Corynebacterium xerosys ATCC 373 and Pseudomonas aeruginosa ATCC 15442. The only methods capable of completely inactivating all strains were thermal methods. Viability and sublethal damage was dependent on the strain, culture medium and lysis or inactivation method. The lysis and inactivation methods did not maintain antimicrobial activity against Cutibacterium acnes ATCC 6919. The SEM of the strains in whey showed organic matter surrounding the cells. The technique of emulsification / internal gelation with 5% sodium alginate provided encapsulation of 61.94% without lyophilization and 37.83% with lyophilization, without gastrointestinal survival. Physical-chemical characteristics were analyzed by FTIR and optical microscope. The microcapsule lost anti-acne activity. DE4 strain optimization was carried out by enriching the whey, using the fractional factorial design (DFF) and a central composite rotational design (DCCR), outlined by the Minitab program18. The bioprocess achieved a high level of cell growth, reaching values greater than 10¹⁵ CFU mL-1. We conclude that LAB in its lysed or inactivated and encapsulated forms did not maintain promising anti-acne activity. The optimization of probiotic strain production makes it increasingly possible to scale up the bioprocess.

Rubella is a disease of epidemiological importance caused by infection with the rubella virus (RV). The virus is transmitted by infected people through airborne droplets. Many cases of congenital anomalies or fetal deaths have been reported after survived by RV during the first 16 weeks of pregnancy. These congenital anomalies are particular to Down syndrome. congenital rubella (CRS). In addition, after birth, the child with CRS may present with deafness, eye disorders and heart diseases, in addition to other late sequelae, including autism, diabetes mellitus and thyroid dysfunction. One of the biggest challenges for the control of Rubella consist in the development of diagnostic strategies that ensured its identification with high accuracy, and, at the same time, can be applied in poor or developing countries. This means that the method the diagnosis necessarily needs to present high performance, reproducibility, in addition to having a low production cost. These premises are fundamental in the incorporation of the diagnostic tool for application to the Unified Health System (SUS), or still intended for private clinical analysis laboratories, resulted in greater diagnostic efficiency and cost reduction. Our group has been working with the strategy of producing innovative proteins, expressed in Escherichia coli (E. coli), which resulted in more sensitive tests with lower production costs. Among the recombinant proteins developed by us group is the recombinant multiepitope chimera called rMERUB, developed by SOUZA et al. (2021), which in preliminary ELISA assays, results proved promising for diagnosing rubella. The present work has with the objective of improving the preliminary results obtained by the authors, in addition to developing new technologies for the production of antigens applied as inputs in the diagnosis by ELISA for Rubella.

Biosurfactants are produced by various microorganisms. The biotechnological properties of biosurfactants (BS) are widely described in the literature. Thus, this work aimed to evaluate the potential of application of the lipopeptide, produced from a microorganism, in face of the bactericidal, fungicidal, effluent decanting properties, in the spreading of motor oil, in the bioremediation of motor oil in the sand, in the bioremediation of heavy metals in soil and plants, in addition to its toxicity in in vivo assays with Artemia salina. As a result, it was demonstrated that the lipid at a concentration of 500 mg/mL presented an inhibition halo of 9 mm and an MIC value of 0.08 mg/mL for the Gran-negative bacterium Aeromonas hidroplyla, while for the filamentous fungus Colletotrichum sp. no inhibitory activity was detected. The qualitative evaluation of the lipopeptide in the treatment of effluents showed the ability to decant the sewage effluent in 24 h, while the one with aluminum sulfate took 30 min. Visual analysis showed that the lipopeptide performed better in the oil dispersion area when compared to Triton X-100 and SDS. The oil scattering ratio was 75, 48.7 and 59.7% for BS, Triton X-100 and SDS, respectively. After 24 h, it was possible to demonstrate that the oil in the presence of BS was concentrated in the center of the plate. Visual analysis of the bioremediation of motor oil in sand showed that the use of lipopeptide at a concentration of 0.1 mg/mL was more efficient than at a concentration of 0.5 mg/mL and in relation to the use of 0.1 mg/mL of SDS. The lipopeptide in the concentration of 0.1 and 0.5 mg/mL showed a recovery rate of motor oil from contaminated sand equivalent to 80 and 23%, respectively. SDS at a concentration of 0.1 mg/mL showed a recovery rate of 64%. At low concentrations (up to 0.1 mg/mL), the supernatants were non-toxic and did not affect swimming behavior or the survival rate of brine shrimp. At high concentrations (0.5 and 1 mg/mL), the mortality rate for the lipopeptide was 10%, while for the SDS compound it was 100%. We conclude that the lipopeptide produced from a microorganism showed a high potential for application in various sectors of the industry, such as the environment and health.

Enterocins are a group of bacteriocins secreted by bacteria of the genus Enterococcus sp., which have antimicrobial, antiviral and antitumor activities well documented in the literature; however, few studies describe the anti-candida activity of these enterocins. Despite their probiotic characteristic, and their wide use in the industry against food spoilage, the feasibility of inserting enterococci in food has been ruled out due to the increased occurrence of virulence factors in these strains. An excellent approach to solving this problem is to express enterocins heterologously. In this work, Enterococcus faecium Enterocin B was characterized by bioinformatics tools, expressed and purified from E. coli BL21(DE3) and ArcticExpress cells in order to verify its solubility during expression. Finally, Enterocin B was tested for its antifungal effect on antifungal resistant Candida albicans and Candida glabrata reference strains. The results showed a difference in the tertiary structure when compared to the literature. The structure analyzed in this work showed two a-helix regions, while in the literature, Enterocin B showed four a-helix regions. However, the results of physicochemical predictions such as amount of amino acids (53), theoretical pI (9.25), molecular weight (5.4 kDa), amphipacity, among others, corroborate with results published by other authors. Large-scale heterologous expression was performed in ArcticExpress cells due to the partially soluble expression of the peptide, because in BL21 (DE3) cells, expression occurred entirely in the form of inclusion bodies. The in vitro results showed that Enterocin B was not able to inhibit the growth of Candida albicans and Candida glabrata strains at concentrations of 250 and 500 µg/mL, respectively, in the MIC assay. A likely hypothesis would be that the GB1 fusion protein, expressed together with Enterocin B to increase the stability and solubility of the peptide, might be preventing the initial interaction that occurs between the N-terminal portion of Enterocin B and the target cell membrane. Thus, additional tests involving Enterocin B, cleaved from the fusion protein, are necessary for a correct conclusion about the activity of this peptide against yeast strains.

Diazepam (DZP) is one of the most well-known benzodiazepines (BZDs), being widely used as an anxiolytic, anticonvulsant and muscle relaxant. Considering the increasing and indiscriminate consumption of anxiolytic drugs worldwide, together with the scarcity of studies that evaluate the effects of BZDs on blood cells, in addition to the fact that, after administration, DZP remains temporarily in the current blood, the aim of the present study was to evaluate the effect of DZP on the Na,K-ATPase and PMCA activities, plasma membrane integrity and parameters related to oxidative stress in human erythrocytes. For this, human blood was incubated with DZP 15 μg/mL (injectable or pure) for 30 and 60 minutes, at room temperature. Subsequently, the following steps were performed: preparation of erythrocyte membrane (ghost), lipid extraction, determination of osmotic fragility, dosage of total phospholipids and cholesterol and evaluation of the activity of antioxidant enzymes and oxidative parameters. Treatment with injectable DZP caused a decrease in the Na, K-ATPase activity, with no significant changes in the other parameters. The group treated with pure DZP showed a greater osmotic fragility when compared to the control group. A 43% increase in membrane phospholipid content was observed after 60 minutes of treatment. The lipid peroxidation index was not altered, but a decrease in H₂O₂ levels and in CAT and AChE activities was observed. In addition, other oxidative stress markers, such as GPx and GSH, were modulated by DZP in a time-dependent manner, and a 46% decrease in Na, K-ATPase activity was observed after 30 minutes of treatment. These results indicate that DZP was able to alter important biochemical parameters in erythrocytes, such as osmotic fragility, lipid content, oxidative stress and ATPasic activity of the membrane in a time-dependent manner. Thus, the several effects observed here reinforce the importance of studies that evaluate the effect of BZDs on erythrocytes, motivating the conscious and controlled use of these drugs and the search for alternative treatments, since the prolonged use of DZP by individuals who are already weakened by diseases that compromise blood cells can significantly affect them.

The Culex genus has great sanitary importance in Brazil, being the transmitter of filariasis and, recently, reported as a possible transmitter of Zika Virus. Widespread throughout the country, the practice of using insecticides is the most commonly employed to control the proliferation of mosquitoes. These insecticides usually cause several damages to the insects, either by altering the activity of some enzymes, such as Acetylcholinesterase (AChE), or in the excessive production of Reactive Oxygen Species (ROS), causing paralysis or even death. For mosquitoes, there are reports that resistance is associated with antioxidant enzymes present in these organisms, such as Superoxide Dismutase (SOD), which protect the animals from damage caused by ROS. Thus, this study aimed to evaluate the effects of insecticides on the oxidative stress pathway and cell membranes in 3rd and 4th instar larvae of Culex spp. To this end, outdoor tanks were set up to obtain the larvae, and then they were exposed to the LC₅₀ of the insecticides. The organophosphate Temephos and the ivermectin derivative Ivomec showed lower LC₅₀, therefore were selected for the analysis of their effects on membrane and oxidative stress of the larvae. The larval groups were exposed to the LC₅₀ of the insecticides for 24 hours and later, material was prepared to perform the evaluation of the Na⁺/K⁺-ATPase pump activity, dosage of GSH, Hydrogen Peroxide, Lipoperoxidation and Carbonylated Protein levels, SOD and AChE enzyme activity. Initially we detected that the amount of total protein decreased after the treatments with the insecticides even though the larvae survived the treatments. Afterwards, we identified in both treatments a significant reduction in SOD activity, followed by a decrease in the amount of Hydrogen Peroxide and GSH. Lipid peroxidation was lower in the treated larvae and carbonylated protein decreased on treatment with Ivomec and increased about 3-fold after treatment with Temephos. Na⁺/K⁺-ATPase and AChE activity were more drastically inhibited after Temephos treatment. The results point to the imbalance caused by the insecticides on the membranes and the function of important proteins such as AChE. Furthermore, the changes in the antioxidant system indicate an attempt to adapt to the changes caused by treatment with the insecticides.

Cardiotonic steroids have been used as cardiac drugs for over 200 years and currently several studies have shown interesting anti-tumor effects with cardiotonic steroids for a number of cell cultures. Cisplatin has played a crucial role in chemotherapy and anticancer treatments for over 30 years. However, treatment with cisplatin can cause serious side effects such as myelosuppression, ototoxicity and nephrotoxicity, in addition to the cellular resistance processes already described in prolonged use of the drug. Studies previously published by our research group have shown a potent synergistic antitumor effect between 1nM digoxin and 1µM cisplatin in cervical cancer (HeLa) cells, and elucidate a possible signaling pathway that would be triggering the antitumor process, where the effect of combined treatment on Na, K-ATPase activity and expression and Src involvement was demonstrated. The aim of the present work is to demonstrate the effect of combined treatment between digoxin and cisplatin on Src activation, through protein expression assays and the effect of combined treatment on oxidative stress parameters in HeLa cells. In addition to demonstrating the effect of treatment on other cell lines, tumor and normal. Combination treatment has been shown to increase phosphorylated Src expression in the activation residue (Tyr416), proving Src involvement in its cytotoxic effect. Analyzing the parameters of oxidative stress, we observed a significant reduction in cellular antioxidant defense mechanisms, SOD activity and GSH content and an increase in 24 hours in H₂O₂ content, confirming the increased cellular oxidative stress across the combined treatment. In addition, the levels of lipid peroxidation and lipid droplet formation were evaluated, where an increase of these parameters was observed after 48 hours of treatment, which is commonly described in cases of oxidative stress. Compared to other cell lines tested, A549 (lung cancer epithelial cells) and WI-38 (normal human fibroblast cells), the combined treatment did not have the ability to significantly alter cell viability compared to control. Our experiments suggest that the antitumor effect of synergistic treatment between digoxin and cisplatin in HeLa cells seems to be correlated with increased in the oxidative stress in this cell type by Na, K-ATPase / Src / ROS activation which in turn could culminating in different cellular effects such as signaling, antiproliferative and apoptotic effects.

The epilepsies are neurological disorders manifested by spontaneous and recurring epileptic seizures, caused by hyper-synchronized neuronal discharges. Temporal lobe epilepsy (TLE) has high prevalence and resistance to drug treatment, with possible evolution causing morphological and molecular alterations in the hippocampus, making surgical intervention necessary to stop the evolution. This work aims to study the morphological parameters and molecular factors related to excitability and apoptosis in hippocampal regions of TLE patients with a right or left onset, who underwent surgery to evaluate new potential neuroprotection targets. Hippocampal morphological evaluation was performed by the volume determination using magnetic resonance imaging and part of the removed tissue during surgery was analyzed by western blot to quantify proteins involved in excitability (Na+, K+-ATPase and GluR1) and the apoptotic pathway (apoptosis inducing factor [AIF], cytochrome c, caspases 8 and 3). Moreover, the correlation coefficient of morphological and molecular parameters was evaluated. The sample was composed of 18 patients, 13 who had surgery on the left side (TLEL) and 5 who had surgery on the right (TLER) side. There was significant decrease in volume when comparing contralateral side of total hippocampus and its regions of TLER patients, except the hippocampal tail, of TLEL patients. Atrophy was observed when comparing volumes of healthy hippocampi and the ictal onset zone of both TLEL and TLER patients and atrophy of contralateral hippocampal tail in ELTE. Protein expression was not significantly different in neither hippocampal head or tail nor when comparing TLEL and TLED patients. Besides, it is suggested that there is a negative correlation between volume and the expression of caspase-3, AIF, cytochrome c, GluR1, α3 and α1 in hippocampal tail. Hippocampal head showed a negative correlation for caspase 3, caspase 8, AIF, cytochrome c, FXYD2, α3 and α1. Caspase 8 showed a positive correlation with tail. GluR1 and α2 did not show correlation with head volume neither do α2 and FXYD2 with head volume. In summary, there is atrophy in the ipsilateral to the ictal onset zone, in addition to the presence of correlation between hippocampal volume and expression of some of the proteins analyzed. However, no molecular difference was observed between hippocampal head and tail in ELTE and ELTD, thus, it isn’t possible to stablish specificity to the laterality or hippocampal regions.

The lipid composition and fluidity of the plasma membrane can be altered by cardiotonic steroids and, consequently, this lipid modulation can directly affect the function of Na, K-ATPase. The interaction of Na, K-ATPase with caveolin-1 has been associated with the maintenance of cholesterol homeostasis in cells. Thus, the aim of this study was to evaluate the involvement of caveolin-1 in the lipid modulation of the plasma membrane caused by cardiotonic steroids. For this, we used a cell line that has caveolae (HeLa cells), and a cell line that doesn’t have this microdomain (Caco-2 cells). We evaluated the effect of treatment with ouabain or 21-BD on the modulation of the content of total phospholipids of these cells, by using colorimetric methods and incorporation of radioactive phosphate. We also evaluated the composition of phospholipids, identifying the content of phosphatidylethanolamine, phosphatidylserine, phosphatidylcholine, phosphatidylinositol, sphingomyelin and lysophosphatidylcholine, by HPTLC. To investigate the effects of treatment with cardiotonic steroids on the modulation of membrane cholesterol, we evaluated the content and distribution of this lipid by Filipin and analyzed the expression of caveolin-1. Ouabain and 21-BD had different effects on the modulation of the lipid profile of the two cell lines. Ouabain had no effect on the total content of membrane phospholipids, although it did occasionally alter some specific phospholipids in the two cell lines. Treatment with 21-BD did not alter the cholesterol content of Caco-2 cell membranes, however, a higher fluorescence indicating a percentage of higher cholesterol in HeLa cells. Futhermore, only 21-BD caused a redistribution of cholesterol to the perinuclear region of HeLa and Caco-2 cells. Caveolin-1 expression didn’t change after treatment with cardiotonic steroids. Therefore, we demonstrate that the cardiotonic steroids, ouabain and 21-BD, caused changes in the lipid composition resulting in a change in the fluidity of the membrane. In addition, we observed changes in the phospholipid content important for different cellular functions. Although cholesterol was redistributed in the two cell lines, an increase in the content of this lipid was observed only in cells that have caveolae, however, caveolin-1 does not seem to be involved in this effect.

Glioblastoma Multiforme (GBM) is the most aggressive type of neoplasm of the central nervous system. Although rare, it has a poor prognosis and a low survival rate. Statistics show an increase in the incidence of cancer with a linear tendency to increase the mortality rate. In addition, GBM does not. Although some treatments already await this type of tumor, they are known to be highly invasive. In this context, an alternative is sought to the range of secondary compounds that natural products can offer, with biological and therapeutic properties. Therefore, the objective of the work was to evaluate an antitumor capacity of derivatives of the species Macairea radula on human glioma cell lines. First, as a positive control, the cytotoxicity of Temozolamide (TMZ) on tumor lines (U251 and GAMG) and on the normal astrocyte line - NHA, was evaluated for 24 and 72 hours. The Tumor Selectivity Index (SI) of the chemotherapeutic agent was calculated. It was observed that TMZ did not show selectivity for any of the studied tumor lines. At the same time, the hexane and chloroform partitions of M. radula were selected through previous screening for fractionation by column chromatography. Six fractions derived from B and 8 derived from C partition were obtained. The cytotoxicity of 12 of the fractions was evaluated, concomitantly with partitions B and C on lines U251, GAMG and NHA (24 and 72 hours). The IS of each of the corresponding extracts was calculated. Those fractions that presented higher IS with 72 hours IC50 were selected - 23-B3, 23-B4, 23-B5 and 23-C6. Characteristic morphological changes, in response to the cytotoxicity of the fractions, were observed. In corroboration, the characterization of cell death was performed through the fluorescence assay. It was observed that there was an increase in the cells in the death stage of the treated cells, with predominantly apoptotic characteristics. Subsequently, the ability of the selected extracts to inhibit the migration of tumor cells was evaluated by the Wound Healing. It was observed that none of the tested fractions was able to inhibit or to reduce the migration of the U251 and GAMG. Accordingly, surface projections were observed, even when the cells were treated. The effect of the fractions on Matrix Metalloproteinase 9 (MMP-9) activity was performed by zimografy. It was observed that fractions 23-B4, 23-B5 and 23-C6 were able to reduce the activity of the proteolytic enzyme. Additionally, these fractions were evaluated on the adhesion of the U251 line. It was possible to notice that the samples modulated the complex involved with the maintenance of the junctions, since they increased cell adhesiveness. In addition, the affinity of Annexin V for phosphatidylserine was tested qualitatively, after treatment of cells (U251) with fractions 23-B4, 23-B5 and 23-C6. It was observed that the number of cells marked with fluorescence was analytically higher when subjected to treatment. In addition, it was possible to verify a higher concentration of markers in the peripheral cell region. Due to the specificity of the test, the data may indicate the induction of cell death by apoptosis. Based on that, these results were used as a protein selection criterion, whose expression was evaluated by Western Blotting. In view of this, the effect of the samples on the modulation of apoptotic enzymes was verified on U251. The 23-B4 fraction did not show the ability to modulate any of the evaluated proteins. Fractions 23-B5 and 23-C6 were able to regulate the expression of proteins involved in the induction of programmed cell death, regardless of Caspase activation, and were shown to activate intrinsic apopototic factors. Finally, the fraction with the highest tumor selectivity (23-B5) was evaluated, in vivo. The fraction was able to significantly reduce the tumor perimeter and it demonstrated potential for the negative modulation of angiogenic factors. The samples were able to modulate parameters involved in tumor progression, being considered promising antitumor candidates.