IMMUNOHISTOCHEMICAL DETECTION OF L CELLS IN GASTROINTESTINAL TRACT MUCOSA OF PATIENTS AFTER SURGICAL TREATMENT FOR CONTROL OF TYPE 2 DIABETES MELLITUS.

OBJECTIVE: Type 2 diabetes mellitus (T2DM) is a disease of global impact that has led to an increase in comorbidities and mortality in several countries. Immunoexpression of the incretin hormones such as glucagon-like peptide-1 (GLP-1) and peptide YY (3-36) (PYY3-36) can be used as a scorer in the gastrointestinal tract to analyze L-cell activity in response to T2DM treatment. This study aimed to investigate the presence, location, and secretion of L cells in the small intestine of patients undergoing the form of bariatric surgery denominated adaptive gastroenteromentectomy with partial bipartition. METHODS: Immunohistochemical assays, quantitative real-time polymerase chain reaction (qPCR), and Western blot analysis were performed on samples of intestinal mucosa from patients with T2DM in both the preoperative and postoperative periods. RESULTS: All results were consistent and indicated basal expression and secretion of GLP-1 and PYY3-36 incretins by L cells. A greater density of cells was demonstrated in the most distal portions of the small intestine. No significant difference was found between GLP-1 and PYY3-36 expression levels in the preoperative and postoperative periods because of prolonged fasting during which the samples were collected. CONCLUSION: The greater number of L cells in activity implies better peptide signaling, response, and functioning of the neuroendocrine system.

Immunohistochemical detection of l cells in gastrointestinal tract mucosa of patients after surgical treatment for control of type 2 diabetes mellitus / Detecção imunohistoquímica de células l na mucosa do trato gastrointestinal de pacientes após tratamento cirúrgico para controle de diabetes mellitus tipo 2

ABSTRACT - BACKGROUND: Type 2 diabetes mellitus (T2DM) is a disease of global impact that has led to an increase in comorbidities and mortality in several countries. Immunoexpression of the incretin hormones such as glucagon-like peptide-1 (GLP-1) and peptide YY (3-36) (PYY3-36) can be used as a scorer in the gastrointestinal tract to analyze L-cell activity in response to T2DM treatment. OBJECTIVE: This study aimed to investigate the presence, location, and secretion of L cells in the small intestine of patients undergoing the form of bariatric surgery denominated adaptive gastroenteromentectomy with partial bipartition. METHODS: Immunohistochemical assays, quantitative real-time polymerase chain reaction (qPCR), and Western blot analysis were performed on samples of intestinal mucosa from patients with T2DM in both the preoperative and postoperative periods. RESULTS: All results were consistent and indicated basal expression and secretion of GLP-1 and PYY3-36 incretins by L cells. A greater density of cells was demonstrated in the most distal portions of the small intestine. No significant difference was found between GLP-1 and PYY3-36 expression levels in the preoperative and postoperative periods because of prolonged fasting during which the samples were collected. CONCLUSION: The greater number of L cells in activity implies better peptide signaling, response, and functioning of the neuroendocrine system.

RESUMO - RACIONAL: O diabetes tipo 2 (DM2) é uma doença de impacto mundial que tem levado ao aumento de comorbidades e mortalidade em vários países. A imunoexpressão dos hormônios incretínicos glp-1 e pyy3-36, pode ser usada como marcador no trato gastrointestinal para analisar a atividade da célula L em resposta ao tratamento do DM2. OBJETIVO: O presente estudo teve como objetivo investigar a presença, localização e secreção de células L no intestino delgado de pacientes submetidos à forma de cirurgia bariátrica denominada gastroenteromentectomia adaptativa com bipartição parcial. MÉTODOS: Ensaios imunohistoquímicos, reação quantitativa em cadeia de polimerase em tempo real (qPCR) e análise de manchas ocidentais foram realizados em amostras de mucosa intestinal de pacientes com diabetes tipo 2 nos períodos pré- e pós-operatório. RESULTADOS: Todos os resultados foram consistentes e indicaram expressão basal e secreção de peptídeos glucagon-1 (GLP-1) e peptídeos YY (PYY3-36) incretinas por células L. Uma maior densidade de células foi demonstrada nas porções mais distais do intestino delgado. Não foi encontrada diferença significativa entre os níveis de expressão GLP-1 e PYY3-36 nos períodos pré-operatório e pós-operatório, provavelmente devido ao estado de jejum prolongado durante o qual as amostras foram coletadas CONCLUSÃO: O maior número de células L em atividade implica melhor sinalização de peptídeo, resposta e funcionamento do sistema neuroendócrino.

Genetic variability in a population of Astyanax scabripinnis: recent bottleneck and the possible influence of individuals with B chromosomes

Access the genetic variability of endangered and isolated populations has become an important conservation tool. Astyanax scabripinnis is a well-known fish model for genetic studies, forming very isolated populations in headwaters. Besides that, this species frequently presents supernumerary chromosomes, which elevates the interest on genetic studies. Genetic diversity of an Astyanax scabripinnispopulation from the Atlantic Forest (Serra da Mantiqueira region, Brazil) was assessed with microsatellite markers for the first time. Since microsatellite markers are not described for this species, we tested markers described for a related species for transferability to A. scabripinnis. Six polymorphic loci were sufficiently reliable for population genetic analysis. We found that this population passed through a recent bottleneck because of the presence of an excess of heterozygotes, low allelic diversity, heterozygosity excess, and small effective population size. Individuals with and without B chromosomes were previously identified in this population and our study found private alleles in the individuals without B chromosomes. Furthermore, when individuals without B chromosomes were removed from the analysis, the population did not present heterozygosity excess, suggesting that the bottleneck event was driven by individuals with B chromosomes. Our results provide an insight into the value of microsatellite markers as molecular tools and is the first genetic study using molecular data of A. scabripinnis from this area.

Patients v. Myriad or the GDPR Access Right v. the EU Database Right.

In 2016, four US cancer patients legally challenged Myriad by claiming full access to all genomic information produced in the course of Myriad's testing of their risks for a variety of cancers. Asserting that Myriad's refusal to provide them with this information violated the HIPAA Privacy Rule, the patients sought a determination of a right to access all their genetic information from testing laboratories. Such access would not only serve their own care, but also enable them to share their genetic data with the scientific community which they alleged Myriad failed to do. A similar case may be brought in Europe under the novel EU GDPR. Specifically, it would put the GDPR right of access to personal data against Myriad's database right under the EU Database Right Directive. The outcome of this case could impact the fate of personalized medicine, which depends on the one hand on patients' having control over their genetic data, and on the other hand on incentives for genetic testing companies to generate these data. We first address the issue of whether the GDPR applies to medical records. We then analyse how GDPR rights could play out in the context of clinical genetic testing and conclude that the GDPR access right stops short of granting unconditional access to all data generated in the process of testing, to the extent that its exercise would result in the violation of medical-professional norms, expose the testing company to potential liability, or compromise normal exploitation of the database of which the personal data form part.

Cocoplum (Chrysobalanus icaco L.) decreases doxorubicin-induced DNA damage and downregulates Gadd45a, Il-1ß, and Tnf-α in vivo.

DNA damage and inflammation are promising targets in disease prevention studies. Since these pathways have shown to be modulated by dietary components, investigating the molecular effects of food becomes relevant. This study aimed at investigating the protective effects of cocoplum (Chrysobalanus icaco L.) against doxorubicin (DXR)-induced damage. Rats were treated with cocoplum (100, 200 or 400mg/kg/day) for 14days, associated or not with DXR (15mg/kg b.w.). Tissue-targeted comet assay and the oxidative stress parameters oxidized/reduced glutathione and catalase were investigated in liver, kidney, and heart. The expressions of DNA damage/repair (Gadd45a, Parp1, Xrcc2) and proinflammatory genes (Il-1ß, Il-6, Nf-κb, Tnf-α) were performed by real-time quantitative PCR. Cocoplum decreased DNA damage and the expressions of Gadd45a, Il-1ß, and Tnf-α induced by DXR. These findings demonstrate that cocoplum fruits possess antigenotoxic and anti-inflammatory effects against DXR-induced damage and encourage other in vivo/clinical studies with this fruit.

Erythrosine B and quinoline yellow dyes regulate DNA repair gene expression in human HepG2 cells.

Erythrosine B (ErB) is a cherry pink food colorant and is widely used in foods, drugs, and cosmetics. Quinoline yellow (QY) is a chinophthalon derivative used in cosmetic compositions for application to the skin, lips, and/or body surface. Previously, ErB and QY synthetic dyes were found to induce DNA damage in HepG2 cells. The aim of this study was to investigate the molecular basis underlying the genotoxicity attributed to ErB and QY using the RT2 Profiler polymerase chain reaction array and by analyzing the expression profile of 84 genes involved in cell cycle arrest, apoptosis, and DNA repair in HepG2 cells. ErB (70 mg/L) significantly decreased the expression of two genes ( FEN1 and REV1) related to DNA base repair. One gene ( LIG1) was downregulated and 20 genes related to ATR/ATM signaling ( ATR, RBBP8, RAD1, CHEK1, CHEK2, TOPB1), nucleotide excision repair ( ERCC1, XPA), base excision repair ( FEN1, MBD4), mismatch repair ( MLH1, MSH3, TP73), double strand break repair ( BLM), other DNA repair genes ( BRIP1, FANCA, GADD45A, REV1), and apoptosis ( BAX, PPP1R15A) were significantly increased after treatment with QY (20 mg/L). In conclusion, our data suggest that the genotoxic mechanism of ErB and QY dyes involves the modulation of genes related to the DNA repair system and cell cycle.

Karyotype analysis of three species of Corydoras (Siluriformes: Callichthyidae) from southern Brazil: rearranged karyotypes and cytotaxonomy

The genus Corydoras comprises a diversity of species with different diploid numbers. We compared cytogenetic data among Corydoras species from different rivers of the Ponta Grossa Arch region in southern Brazil. Corydoras ehrhardti and C. aff. paleatus have a similar karyotype formula and the same diploid number (2n = 44). Corydoras lacrimostigmata has a higher diploid number, with 2n = 58 chromosomes. Fluorescence in situ hybridization using 5S and 18S ribosomal DNA probes suggests that these ribosomal DNA sequences are involved in chromosomal rearrangements in these Corydoras species. 5S rDNA is a chromosomal marker that is considered to be unique to the species analyzed in this study. Signals of interstitial telomeric sites are seen in a chromosome pair of C. lacrimostigmata, suggesting chromosomal rearrangements via fusions or translocations. This study revealed that C. ehrhardti and C. aff. paleatus have exclusive chromosomal markers associated with chromosome differentiation, which we speculate to prevent genetic introgression.(AU)

Corydoras compreende um gênero diversificado com espécies de diferentes números diploides. Nós comparamos dados citogenéticos de espécies de Corydoras de diferentes rios da região do Arco de Ponta Grossa no sul do Brasil. Corydoras ehrhardti e C. aff. paleatus tem fórmula cariotípica similar e o mesmo número diploide (2n = 44). Corydoras lacrimostigmata tem um número diploide maior, com 2n= 58 cromossomos. A hibridação in situ fluorescente (FISH) com sondas de DNA ribossomal 5S e 18S sugere que estas sequências de DNA ribossomal estão envolvidas em rearranjos cromossômicos nestas espécies de Corydoras. A marcação do DNAr 5S foi considerada espécie-específico para as espécies analisadas neste estudo. Sinais de sítios teloméricos intersticiais foram vistos em um par de cromossomos de C. lacrimostigmata sugerindo a ocorrência de rearranjos cromossômicos como fusões ou translocações. Este estudo revelou que as espécies C. ehrhardti e C. aff. paleatus têm marcadores cromossômicos exclusivos associados à diferenciação cromossômica, os quais, em nossa hipótese, podem prevenir a introgressão gênica.(AU)

Chrysobalanus icaco L. fruits inhibit NADPH oxidase complex and protect DNA against doxorubicin-induced damage in Wistar male rats.

Chrysobalanus icaco L. is an underexplored plant found in tropical areas around the globe. Currently, there is no apparent information regarding the effects C. icaco fruits may exert in vivo or potential role in health promotion. This study aimed at providing evidence regarding the in vivo influence of this fruit on antigenotoxicity, antimutagenicity, and oxidative stress in rats. Male Wistar rats were treated with 100, 200, or 400 mg/kg body weight (bw)/d C. icaco fruit for 14 d. Doxorubicin (DXR, 15 mg/kg bw, ip) was used for DNA damaging and as an oxidant to generate reactive oxygen species (ROS). Genomic instability was assessed by the comet assay and micronucleus (MN) test, while antioxidant activity was determined by oxidative burst of neutrophils. Chrysobalanus icaco fruit polyphenols were quantified and characterized by high-performance liquid chromatography coupled to a diode array detector and tandem mass spectrometer (HPLC-DAD-MS/MS). The concentrations of 19 chemical elements were determined by inductively coupled plasma-mass spectroscopy (ICP-MS). Significant amounts of polyphenols, magnesium, and selenium were found in C. icaco fruit. This fruit displayed in vivo antioxidant activity against DXR-induced damage in rat peripheral blood neutrophils, antigenotoxicity in peripheral blood cells, and antimutagenicity in bone-marrow cells and peripheral blood cells. Correlation analyses between endpoints examined indicated that the mechanism underlying chemopreventive actions of C. icaco fruit was attributed to inhibition of NADPH oxidase complex manifested as low levels of DNA damage in animals exposed to DXR. Data indicate that phytochemicals and minerals in C. icaco fruit protect DNA against damage in vivo associated with their antioxidant properties.

Differential genotoxicity and cytotoxicity of phomoxanthone A isolated from the fungus Phomopsis longicolla in HL60 cells and peripheral blood lymphocytes.

Phomoxanthone A (PhoA) is a compound isolated from the endophytic fungus Phomopsis longicolla, associated with marine algae Bostrychia radicans. Although this metabolite was previously described regarding its high biological potential, there are no reports concerning the effects of this compound on DNA integrity. This study aimed to evaluate, in lymphocytes and promyelocytic leukemia HL60 cells, the cytotoxicity of this compound through MTT and neutral red (NR) assays, as well as its genotoxicity and mutagenicity by alkaline comet assay and cytokinesis-block micronucleus cytome assay (CBMN-Cyt), respectively. Cells were treated with PhoA concentrations ranging from 0.01 to 100.0µg/mL, and the results show that this molecule did not exhibit cytotoxicity, genotoxicity or mutagenicity in lymphocytes at any tested concentration. Furthermore, PhoA was highly cytotoxic, genotoxic and mutagenic to HL60 cells, establishing a differential response of this natural product in normal and cancer cells. PhoA was highly selective towards HL60 compared to lymphocytes, causing no damage in the latter cell line, suggesting that this compound could be a promising compound in antitumoral drug development.

Swim training and the genetic expression of adipokines in monosodium glutamate-treated obese rats.

OBJECTIVE: The aim of this study was to evaluate the genetic expression of adipokines in the adipocytes of monosodium glutamate (MSG)-treated obese rats submitted to physical activity. MATERIALS AND METHODS: Obesity was induced by neonatal MSG administration. Exercised rats (MSG and control) were subjected to swim training for 30 min for 10 weeks, whereas their respective controls remained sedentary. Total RNA was obtained from sections of the mesenteric adipose tissue of the rats. mRNA levels of adiponectin (Adipoq), tumor necrosis factor alpha (Tnf), peroxisome proliferator-activated receptor alpha (Ppara), and peroxisome proliferator-activated receptor gamma (Pparg) adipokines were quantified by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). RESULTS: In the exercise-trained control group, the expression of Adipoq increased compared to the sedentary control, which was not observed in the MSG-obese rats. Increased levels of Tnf in MSG-obese rats were not reversed by the swim training. The expression of Ppara was higher in sedentary MSG-obese rats compared to the sedentary control. Swimming increased this adipokine expression in the exercise-trained control rats compared to the sedentary ones. mRNA levels of Pparg were higher in the sedentary MSG-rats compared to the sedentary control; however, the exercise did not influenced its expression in the groups analyzed. CONCLUSIONS: In conclusion, regular physical activity was not capable to correct the expression of proinflammatory adipokines in MSG-obese rat adipocytes.

Swim training and the genetic expression of adipokines in monosodium glutamate-treated obese rats

Objective The aim of this study was to evaluate the genetic expression of adipokines in the adipocytes of monosodium glutamate (MSG)-treated obese rats submitted to physical activity.Materials and methods Obesity was induced by neonatal MSG administration. Exercised rats (MSG and control) were subjected to swim training for 30 min for 10 weeks, whereas their respective controls remained sedentary. Total RNA was obtained from sections of the mesenteric adipose tissue of the rats. mRNA levels of adiponectin (Adipoq), tumor necrosis factor alpha (Tnf), peroxisome proliferator-activated receptor alpha (Ppara), and peroxisome proliferator-activated receptor gamma (Pparg) adipokines were quantified by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR).Results In the exercise-trained control group, the expression of Adipoq increased compared to the sedentary control, which was not observed in the MSG-obese rats. Increased levels of Tnf in MSG-obese rats were not reversed by the swim training. The expression of Ppara was higher in sedentary MSG-obese rats compared to the sedentary control. Swimming increased this adipokine expression in the exercise-trained control rats compared to the sedentary ones. mRNA levels of Pparg were higher in the sedentary MSG-rats compared to the sedentary control; however, the exercise did not influenced its expression in the groups analyzed.Conclusions In conclusion, regular physical activity was not capable to correct the expression of proinflammatory adipokines in MSG-obese rat adipocytes.

Hipertensão arterial de difícil controle e o polimorfismo A1166C do gene AT1 / Difficult-to-treat hypertension and AT1 A1166C gene polymorphism

Fundamentos: Polimorfismos presentes em genes que codificam proteínas do sistema renina-angiotensina aldosterona (SRAA) estão associados com o quadro de hipertensão arterial sistêmica (HAS) em algumas populações. Estudos demonstram a relação entre o polimorfismo A1166C no gene do receptor tipo 1 da angiotensina II (AT1) com a HAS, mas os dados ainda são controversos. Objetivo: Analisar a presença deste polimorfismo em pacientes portadores de HAS resistente da região dos Campos Gerais (PR), Brasil. Materiais e Métodos: Grupos de pacientes com hipertensão de fácil (G1) (n = 34) e difícil controle medicamentoso (G2) (n = 39) foram analisados quanto ao polimorfismo mencionado, utilizando-se a técnica de Polymerase Chain Reaction­ Restriction Fragment Lenght Polymorphism (PCR-RFLP). Os pacientes foram categorizados em três genótipos: AA, AC e CC. As frequências alélicas e genotípicas foram calculadas para cada grupo e os dados confrontados com as características metabólicas e antropométricas dos indivíduos. Resultados: não houve diferença entre os grupos quanto a sexo e idade. O índice de massa corporal (IMC), pressão arterial sistólica (PAS), diastólica (PAD) e número de anti-hipertensivos utilizados foram maiores no G2. Asfrequências alélicas e genotípicas mostraram-se semelhantes entre os grupos (p > 0,05). Conclusão: Nesta população, este polimorfismo não está associado ao fácil ou difícil controle da pressão arterial (PA). Possivelmente, outros fatores devem estar influenciando a HAS nestes pacientes

Background: Polymorphisms in genes encoding proteins of the renin-angiotensin-aldosterone system (RAAS) are associated with systemic arterial hypertension (SAH) in some populations. Some reports demonstrated the relationship between the angiotensin II type 1 receptor (AT1) A1166C gene polymorphism with SAH, but the data are still controversial. Objective: To analyze the presence of this polymorphism in patients porting difficult-to-treat SAH from Campos Gerais region (PR), Brazil. Materials and Methods: Groups of patients porting hypertension easy (G1) (n = 34) and difficult-to-treat using drugs (G2) (n = 39) were analyzed according to the polymorphism mentioned, using the Polymerase Chain Reaction­ Restriction Fragment Lenght Polymorphism (PCR-RFLP) technique. The patients were categorized into three genotypes: AA, AC and CC. The allele and genotype frequencies were calculated and the results were compared with metabolic and anthropometric characteristics of the patients. Results: There was no difference between groups regarding gender and age. The body mass index (BMI), systolic and diastolic blood pressures and the number of antihypertensive drugs were higher in G2. The allele and genotype frequencies were similar between the groups (p > 0.05). Conclusions: In this population, the polymorphism analyzed is not associated with easy or difficult-to-treat SAH. Possibly, other factors are influencing the hypertension in these patients

Evidence of incipient speciation in astyanax scabripinnis species complex (teleostei: characidae)

Two populations of the Astyanax scabripinnis complex, isolated by a waterfall with over 100 meters depth and inhabiting different altitudes of the same river (1850 m a.s.l. and 662 m a.s.l.) were compared in reproductive data, geometric morphometry, tooth morphology, anal-fin rays counts, and karyotype, in order to test the hypothesis of speciation between the two populations. The results in the geometric morphometry analysis showed differences between the populations. Discriminant function analysis (DFA) and canonical variance analysis revealed sexual dimorphism. Secondary sexual characters, such as hooks in the anal fin rays of the males are absent in the lower altitude population. Both populations had the same macro karyotype structure, except for the absence of B chromosomes in the lower altitude population. The fluorescence in situ hybridization showed differences for both markers (18S rDNA and 5S rDNA), and reproductive data suggests pre-zygotic reproductive isolation among the two populations. The data showed the absence of gene flow, indicating that an incipient speciation process has occurred, which leads the two populations to follow independent evolutionary pathways.

Duas populações do complexo Astyanax scabripinnis isoladas por uma queda d´água de mais de 100 metros de altura e localizadas em diferentes altitudes do mesmo rio (662 m e 1850 m a.s.l.) foram comparadas através de dados de reprodução, cariótipo, morfometria geométrica, morfologia dentária, e número de raios da nadadeira anal, de modo a testar a hipótese de especiação entre as duas populações. Os resultados de morfometria geométrica mostraram diferenças entre as populações. A análise da função discriminante (DFA) e a análise de variância canônica (CVA) demonstraram a presença de dimorfismo sexual. Caracteres sexuais secundários, como ganchos em raios da nadadeira anal dos machos, estão ausentes na população de menor altitude. Ambas as populações têm a mesma macro estrutura cariotípica, exceto pela ausência de cromossomos B na população de menor altitude. A hibridação in situ mostrou diferenças para ambos os marcadores (rDNA 18S e rDNA 5S), e os dados de reprodução sugerem isolamento reprodutivo pré-zigótico entre as duas populações. Os dados mostram ausência de fluxo gênico, indicando que ocorreu um processo de especiação incipiente, o que leva as duas populações seguirem vias evolutivas independentes.

Species with medicinal and mystical-religious uses in São Francisco do Conde, Bahia, Brazil: a contribution to the selection of species for introduction into the local Unified Health System

We investigated the knowledge and practices of local residents in São Francisco do Conde, Bahia, regarding the use of medicinal and mystical plants with the aim of proposing strategies for the incorporation of phytotherapies into the local Unified Health System through local Basic Health Clinics. This municipality was founded during the early colonization of Brazil, introducing the monoculture of sugarcane and slave labor to the region, resulting in a currently largely Afro-Brazilian population. Key informants and local specialists were interviewed and workshops were undertaken at the Basic Health Clinics to collect data and information. The interviewees made 254 references to 126 plant species distributed among 107 genera and 50 families. Among the species cited with medicinal or mystical uses, 51.6% were considered autochtonous, and 42.8% were cited in at least one document of the Brazilian Health Ministry; of these, 11.1% were mentioned in four to eight documents, indicating potential for introduction to the local Unified Health System. The valorization of local knowledge and practices concerning the use of medicinal plants represents an important approach to public health efforts.

Relação do polimorfismo C825T do gene GNB3 e hipertensão arterial sistêmica de difícil controle / Relationship between C825T polymorphism of the GNB3 gene and dificult-to-treat hypertension

Fundamentos: O polimorfismo C825T do gene GNB3 está associado à hipertensão arterial sistêmica (HAS) em algumas populações já analisadas, porém alguns estudos se mostram controversos no que se refere a esta relação. Objetivo: Avaliar a relação do polimorfismo C825T do gene GNB3 com a HAS de difícil controle medicamentoso em hipertensos de Campos Gerais, PR - Brasil. Métodos: Em relação ao polimorfismo C825T de GNB3, foram determinados os genótipos de 60 hipertensos, os quais foram estratificados em dois grupos (fácil e difícil controle medicamentoso), por meio da técnica de PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Lenght Polymorphism). Foram avaliadas as frequências alélicas e genotípicas, utilizando-se o teste do qui-quadrado de Pearson, com correção de Yates e odds ratio (OR). Resultados: Não houve diferenças entre os grupos, quando comparadas as frequências alélicas e genotípicas, indicando que a população está em equilíbrio. A probabilidade de o paciente possuir o polimorfismo e a HAS de difícil controle foi 53,5 % (OR=1,15; IC95 % = 0,41-3,26), analisando-se os genótipos. Já a análise dos alelos, separadamente, mostrou uma associação de 55,4 % (OR=1,24; IC95 % = 0,59-2,57). Conclusão: Nesta população não foi encontrada relação entre o polimorfismo C825T do gene GNB3 e a HAS de difícil controle, indicando que outros fatores estão influenciando a manifestação dessa doença nestes pacientes.

Background: C825T polymorphism of the GNB3 gene is associated with systemic arterial hypertension (SAH) in some studied populations, although certain studies are controversial in terms of this relationship. Objective: To evaluate the relationship between C825T polymorphism of the GNB3 gene and difficult-to-treat SAH among hypertensive patients in Campos Gerais, Paraná State, Brazil. Methods: With regard to C825T polymorphism of the GNB3 gene, the genotypes were defined for sixty hypertensive patients divided in 2 groups (easy and difficult-to-treat with drugs), using the Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) technique. The allele and genotype frequencies were assessed through the Pearson chi-square test, with Yates correction and odds ratio (OR). Results: There were no differences between the groups when comparing the allele and genotype frequencies, indicating that the population is in equilibrium. The probability that a patient has polymorphism with difficult-to-treat SAH reached 53.5% (OR=1.15, 95%CI = 0.41-3.26), analyzing the genotypes. A separate allele analysis showed an association of 55.4% (OR=1.24, 95%CI = 0.59-2.57). Conclusion: No relationship was found in this population between C825T polymorphism of the GNB3 gene and difficult-to-treat SAH, indicating that other factors are influencing the appearance of this disease among these patients.

Evaluation of the antihypertensive properties of yellow passion fruit pulp (Passiflora edulis Sims f. flavicarpa Deg.) in spontaneously hypertensive rats.

Various species of the genus Passiflora have been extensively used in traditional medicine as sedatives, anxiolytics, diuretics and analgesics. In the present study, after the identification and quantification of phytochemical compounds from yellow passion fruit pulp by liquid chromatography-photodiode array-mass spectrometry (HPLC-PDA-MS/MS), its antihypertensive effect was investigated on spontaneously hypertensive rats. Additionally, the renal function, evaluated by kidney/body weight, serum creatinine, proteinuria, urinary flow, reduced glutathione (GSH) levels and thiobarbituric acid-reactive substances (TBARS) and mutagenicity in bone marrow cells were assessed to evaluate the safety of passion fruit consumption. Yellow passion fruit pulp (5, 6 or 8 g/kg b.w.) was administered by gavage once a day for 5 consecutive days. HLPC-PDA-MS/MS analysis revealed that yellow passion fruit pulp contains phenolic compounds, ascorbic acid, carotenoids and flavonoids. The highest dose of passion fruit pulp significantly reduced the systolic blood pressure, increased the GSH levels and decreased TBARS. There were no changes in renal function parameters or the frequency of micronuclei in bone marrow cells. In conclusion, the antihypertensive effect of yellow passion fruit pulp, at least in part, might be due to the enhancement of the antioxidant status. The exact mechanisms responsible by this effect need further investigation.

Overtraining is associated with DNA damage in blood and skeletal muscle cells of Swiss mice.

BACKGROUND: The alkaline version of the single-cell gel (comet) assay is a useful method for quantifying DNA damage. Although some studies on chronic and acute effects of exercise on DNA damage measured by the comet assay have been performed, it is unknown if an aerobic training protocol with intensity, volume, and load clearly defined will improve performance without leading to peripheral blood cell DNA damage. In addition, the effects of overtraining on DNA damage are unknown. Therefore, this study aimed to examine the effects of aerobic training and overtraining on DNA damage in peripheral blood and skeletal muscle cells in Swiss mice. To examine possible changes in these parameters with oxidative stress, we measured reduced glutathione (GSH) levels in total blood, and GSH levels and lipid peroxidation in muscle samples. RESULTS: Performance evaluations (i.e., incremental load and exhaustive tests) showed significant intra and inter-group differences. The overtrained (OTR) group showed a significant increase in the percentage of DNA in the tail compared with the control (C) and trained (TR) groups. GSH levels were significantly lower in the OTR group than in the C and TR groups. The OTR group had significantly higher lipid peroxidation levels compared with the C and TR groups. CONCLUSIONS: Aerobic and anaerobic performance parameters can be improved in training at maximal lactate steady state during 8 weeks without leading to DNA damage in peripheral blood and skeletal muscle cells or to oxidative stress in skeletal muscle cells. However, overtraining induced by downhill running training sessions is associated with DNA damage in peripheral blood and skeletal muscle cells, and with oxidative stress in skeletal muscle cells and total blood.

Methionine concentration in the diet has a tissue-specific effect on chromosomal stability in female mice.

Inadequate nutrient intake can influence the genome. Since methionine is an essential amino acid that may influence DNA integrity due to its role in the one-carbon metabolism pathway, we were interested in whether methionine imbalance can lead to genotoxic events. Adult female Swiss mice were fed a control (0.3% dl-methionine), methionine-supplemented (2.0% DL-methionine) or methionine-deficient (0% DL-methionine) diet over a 10-week period. Chromosomal damage was assessed in peripheral blood using a micronucleus test, and DNA damage was assessed in the liver, heart and peripheral blood tissues using a comet assay. The mRNA expression of the mismatch repair genes Mlh1 and Msh2 was analyzed in the liver. The frequency of micronucleus in peripheral blood was increased by 122% in the methionine-supplemented group (p<0.05). The methionine-supplemented diet did not induce DNA damage in the heart and liver tissues, but it increased DNA damage in the peripheral blood. The methionine-deficient diet reduced basal DNA damage in liver tissue. This reduction was correlated with decreased mRNA expression of Msh2. Our results demonstrate that methionine has a tissue-specific effect because methionine-supplemented diet induced both chromosomal and DNA damage in peripheral blood while the methionine-deficient diet reduced basal DNA damage in the liver.

In vivo genotoxicity and oxidative stress evaluation of an ethanolic extract from piquiá (Caryocar villosum) pulp.

In this study, the ethanolic extract obtained from piquiá pulp was assessed for genotoxicity and oxidative stress by employing the micronucleus test in bone marrow and peripheral blood cells in addition to comet, thiobarbituric-acid-reactive substances (TBARS), and reduced glutathione assays in the liver, kidney, and heart. Additionally, phytochemical analyses were performed to identify and quantify the chemical constituents of the piquiá extract. Wistar rats were treated by gavage with an ethanolic extract from piquiá pulp (75 mg/kg body weight) for 14 days, and 24 h prior to euthanasia, they received an injection of saline or doxorubicin (15 mg/kg body weight, intraperoneally). The results demonstrated that piquiá extract at the tested dose was genotoxic but not mutagenic, and it increased the TBARS levels in the heart. Further studies are required to fully elucidate how the properties of ethanolic extract of piquiá pulp can affect human health.

Genotoxicity assessment of Copaiba oil and its fractions in Swiss mice.

Copaiba oil-resin, extracted from the trunk of Copaifera, and traditionally used in folk medicine in the treatment of various disorders, has been shown to be an effective antiinflamatory, antitumor, antitetanus, antiseptic and anti-blenorrhagea agent. As, there are few studies evaluating its genotoxicity, this aspect of the commercial oil-resin, and its volatile and resinous fractions, were evaluated in mice by comet assay and micronucleus (MN) test. A single dose of oil resin, volatile or resin fractions (500; 1,000 or 2,000 mg/kg b.w.) was administered by gavage. The chemical compositions of Copaiba oil resin and its fractions was analyzed by gas chromatography. According to comet assaying, treatment with either one did not increase DNA damage, and as to MN testing, there was no alteration in the incidence of micronucleated polychromatic erythrocytes. Chromatographic analysis of the oil-resin itself revealed sesquiterpenes, diterpenic carboxylic acid methyl esters and high levels of ß-caryophyllene. Thus, it can be assumed that the oil resin and volatile and resinous fractions from the commercial product are not genotoxic or mutagenic.