MGMT and MLH1 methylation in Helicobacter pylori-infected children and adults

AIM: To evaluate the association between Helicobacter pylori(H. pylori) infection and MLH1 and MGMT methylation and its relationship with microsatellite instability (MSI). METHODS: The methylation status of the MLH1 and MGMT promoter region was analysed by methylation specific methylation-polymerase chain reaction (MSPPCR) in gastric biopsy samples from uninfected or H. pylori -infected children (n = 50), from adults with chronic gastritis (n = 97) and from adults with gastric cancer (n = 92). MLH1 and MGMT mRNA expression were measured by real-time PCR and normalised to a constitutive gene (β actin). MSI analysis was performed by screening MSI markers at 4 loci (Bat-25, Bat-26, D17S250 and D2S123) with PCR; PCR products were analysed by single strand conformation polymorphism followed by silver staining. Statistical analyses were performed with either the χ 2 test with Yates continuity correction or Fisher's exact test, and statistical significance for expression analysis was assessed using an unpaired Student's t -test. RESULTS: Methylation was not detected in the promoter regions of MLH1 and MGMT in gastric biopsy samples from children, regardless of H. pylori infection status. The MGMT promoter was methylated in 51% of chronic gastritis adult patients and was associated with H. pylori infection (P < 0.05); this region was methylated in 66% of gastric cancer patients, and the difference in the percentage of methylated samples between these patients and those from H. pylori -infected chronic gastritis patients was statistically significant (P < 0.05). MLH1 methylation frequencies among H. pylori -infected and non-infected chronic gastritis adult patients were 13% and 7%, respectively. We observed methylation of the MLH1 promoter (39%) and increased MSI levels (68%) in samples from gastric cancer patients in comparison to samples from H. pylori -infected adult chronic gastritis patients (P < 0.001 and P < 0.01, respectively). The frequency of promoter methylation for both genes was higher in gastric cancer samples than in H. pylori -positive chronic gastritis samples (P < 0.05). The levels of MLH1 and MGMT mRNA were significantly reduced in chronic gastritis samples that were also hypermethylated (P < 0.01). MGMT promoter region was analysed by methylation specific methylation-polymerase chain reaction (MSPPCR) in gastric biopsy samples from uninfected or H. pylori -infected children (n = 50), from adults with chronic gastritis (n = 97) and from adults with gastric cancer (n = 92). MLH1 and MGMT mRNA expression were measured by real-time PCR and normalised to a constitutive gene (β actin). MSI analysis was performed by screening MSI markers at 4 loci (Bat-25, Bat-26, D17S250 and D2S123) with PCR; PCR products were analysed by single strand conformation polymorphism followed by silver staining. Statistical analyses were performed with either the χ 2 test with Yates continuity correction or Fisher's exact test, and statistical significance for expression analysis was assessed using an unpaired Student's t -test. RESULTS: Methylation was not detected in the promoter regions of MLH1 and MGMT in gastric biopsy samples from children, regardless of H. pylori infection status. The MGMT promoter was methylated in 51% of chronic gastritis adult patients and was associated with H. pylori infection (P < 0.05); this region was methylated in 66% of gastric cancer patients, and the difference in the percentage of methylated samples between these patients and those from H. pylori -infected chronic gastritis patients was statistically significant (P < 0.05). MLH1 methylation frequencies among H. pylori -infected and non-infected chronic gastritis adult patients were 13% and 7%, respectively. We observed methylation of the MLH1 promoter (39%) and increased MSI levels (68%) in samples from gastric cancer patients in comparison to samples from H. pylori -infected adult chronic gastritis patients (P < 0.001 and P < 0.01, respectively). The frequency of promoter methylation for both genes was higher in gastric cancer samples than in H. pylori -positive chronic gastritis samples (P < 0.05). The levels of MLH1 and MGMT mRNA were significantly reduced in chronic gastritis samples that were also hypermethylated (P < 0.01). CONCLUSION: In summary, MGMT and MLH1 methylation did not occur in earlier-stage H. pylori infections and thus might depend on the duration of infection. © 2013 Baishideng. All rights reserved.

Insulin concentrations in cerebellum and body balance in diabetic male rats: Aerobic training effects

Brain insulin has had widespread metabolic, neurotrophic, and neuromodulatory functions and has been involved in the central regulation of food intake and body weight, learning and memory, neuronal development, and neuronal apoptosis. Purpose: The present study investigated the role of swimming training on cerebral metabolism on insulin concentrations in cerebellum and the body balance performance of diabetic rats. Methods: Forty Male Wistar rats were divided in four groups: sedentary control (SC), trained control (TC), sedentary diabetic (SD), and trained diabetic (TD). Diabetes was induced by alloxan (32 mg kg b.w.), single dose injection. The mean blood glucose of diabetic groups was 367 ± 40 mg/dl. Training program consisted in swimming 5 days/week, 1 h/day, 8 weeks, supporting a workload corresponding to 90% of maximal lactate steady state (MLSS). For the body balance testing rats were trained to traverse for 5 min daily for 5-7 days. All dependent variables were analyzed by one-way analysis of variance (ANOVA) and a significance level of p < 0.05 was used for all comparisons. Results: The body balance testing scores were different between groups. Insulin concentrations in cerebellum were not different between groups. Conclusion: It was concluded that in diabetic rats, aerobic training does not induce alterations on cerebellum insulin but induces important metabolic, hormonal and behavioral alterations which are associated with an improvement in glucose homeostasis, serum insulin concentrations and body balance. © 2013 Elsevier Inc.

Effect of physical training on metabolic and bone profile in weaning rats

Introduction: The practice of moderate-intensity exercise can reduce the risk of infections and improve metabolic aspects of the person. Objective: To investigate the effects of aerobic physical training on endocrine and metabolic aspects, bone and immune systems. Methods: Twenty Wistar rats were divided in two groups: sedentary (SG) and trained group (TG). Training program consisted in swimming, 6 weeks, supporting a workload corresponding to 5% of body weight. At the end of the experiment, were performed counting total and differential leukocyte count and hematocrit. After training period, were analyzed glucose, total protein, triglycerides, cholesterol, liver and muscle samples for the determination of the levels of glycogen, and determination of the tibia length and bone area. All dependent variables were analyzed by one-way analysis of variance (ANOVA) and a significance level of P < 0.05 was used for all comparisons. Results: Hematocrit (%) analyzed showed a significant difference, with higher values ffor TG (54.63 ± 1.41) than for the SG (49.5 ± 1.65). The total leukocyte count was not significantly different, as there was no difference in the differential count. Total cholesterol showed significant decrease in TG (TG = 68.27 ± 13.71 mg/dL; SG = 94.44 ± 28.09), the total protein levels also showed significant reduction (TG = 7.3 ± 0.40 g/dL; SG = 7.74 ± 0.36 g/dL) glucose levels and triglyceride showed no significant differences. The bone length showed significant difference (TG = 40±0.14 mm; SG = 42.10 ± 0.12mm). Tibial area showed a lower value for TG (1.53 ± 0.12cm2) than for SG (1.67 ± 0.18cm2); however, the difference was not statistically significant. Conclusion: It can be concluded that aerobic exercise training is able to produce some unique physiological changes in young rats. There is also the need to prescribe exercises that meet the particular maturational stage of development.

Temporomandibular disorders, sleep bruxism, and primary headaches are mutually associated.

To investigate the association among temporomandibular disorders (TMD), sleep bruxism, and primary headaches, assessing the risk of occurrence of primary headaches in patients with or without painful TMD and sleep bruxism. The sample consisted of 301 individuals (253 women and 48 men) with ages varying from 18 to 76 years old (average age of 37.5 years). The Research Diagnostic Criteria for Temporomandibular Disorders were used to classify TMD. Sleep bruxism was diagnosed by clinical criteria proposed by the American Academy of Sleep Medicine, and primary headaches were diagnosed according to the International Classification of Headache Disorders-II. Data were analyzed by chi-square and odds ratio tests with a 95% confidence interval, and the significance level adopted was .05. An association was found among painful TMD, migraine, and tension-type headache (P < .01). The magnitude of association was higher for chronic migraine (odds ratio = 95.9; 95% confidence intervals = 12.51-734.64), followed by episodic migraine (7.0; 3.45-14.22) and episodic tension-type headache (3.7; 1.59-8.75). With regard to sleep bruxism, the association was significant only for chronic migraine (3.8; 1.83-7.84). When the sample was stratified by the presence of sleep bruxism and painful TMD, only the presence of sleep bruxism did not increase the risk for any type of headache. The presence of painful TMD without sleep bruxism significantly increased the risk in particular for chronic migraine (30.1; 3.58-252.81), followed by episodic migraine (3.7; 1.46-9.16). The association between painful TMD and sleep bruxism significantly increased the risk for chronic migraine (87.1; 10.79-702.18), followed by episodic migraine (6.7; 2.79-15.98) and episodic tension-type headache (3.8; 1.38-10.69). The association of sleep bruxism and painful TMD greatly increased the risk for episodic migraine, episodic tension-type headache, and especially for chronic migraine.

Effect of sex and seasons of the year on hematologic and serum biochemical variables of captive brown brocket deer (Mazama gouazoubira)

The Brown brocket deer (Mazama gouazoubira) is the most common free-living and captive deer in South America, especially in Brazil, and has great ecological and scientific significance. However, data on hematological and biochemical parameters in brown brocket deer are scarce. The goal of this study was to establish reference ranges for hematological and biochemical parameters of Mazama gouazoubira, comparing differences during the seasons of the year and between sex. Blood samples from ten adult healthy brown brocket deer (6 female and 4 male) were collected during daytime, monthly, during 12 months. The animals were maintained in individual stable, protected from noise and fed ad libitum with commercial ration and green fodder. For blood collection, animals were submitted to physical restrain for no longer than 2 minutes. The following parameters were determined: red blood cell count (RBC), haemoglobin concentration, packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), white blood cell count (WBC), platelet count, enzyme activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (GGT) and serum levels of alkaline phosphatase (ALP), creatine kinase (CK), total protein (TP), albumin, cholesterol, total calcium, ionic calcium, sodium, potassium, magnesium, triglycerides, creatinine and urea. Values were compared according to season and sex. RBC count, WBC count and MCV suggested seasonal influence. Haemoglobin concentration, PCV and MCV were influenced by sex. Serum concentration of total calcium, ionic calcium, sodium, potassium and magnesium were influenced by season. Serum magnesium was also influenced by sex. The blood parameters herein reported may be useful as reference values for diagnostic and prognostic purposes in captive brown-brocket deer.

Association between bone mass and functional capacity among elderly people aged 80 years and over

Objetivo:Analisar a associação entre a massa óssea e capacidade funcional de idosos com 80 anos ou mais.Métodos:A amostra foi composta por 93 idosos entre 80 e 91 anos (83,2 ± 2,5 anos), 61 mulheres (83,3 ± 2,7 anos) e 32 homens (83,1 ± 2,2 anos) da cidade de Presidente Prudente. A avaliação da massa óssea foi feita pela absorptiometria de dupla energia de raios X (DXA), na qual foram mensurados os valores de conteúdo mineral ósseo (BMC) e densidade mineral óssea (BMD) do fêmur e da coluna (L1-L4). A capacidade funcional foi avaliada por meio dos testes de velocidade para caminhar, equilíbrio estático e força de membros inferiores contidos no questionário Saúde, Bem-Estar e Envelhecimento (Sabe). As variáveis da massa óssea e capacidade funcional foram categorizadas de acordo com os valores de mediana e a pontuação obtida nos testes, respectivamente. Para tratamento estatístico fez-se o teste qui-quadrado, o software usado foi SPSS (13.0) e o nível de significância estabelecido foi de 5%.Resultados:Os idosos do sexo masculino com maior desempenho nos testes funcionais apresentaram maiores valores de BMC de fêmur comparados com os de menor desempenho, resultado não encontrado quando avaliadas as mulheres.Conclusão:Dessa forma, a massa óssea do fêmur para idosos longevos do sexo masculino está associada à capacidade funcional. A avaliação constante da massa mineral óssea e a prática de atividade física ao longo da vida seriam medidas para prevenção das quedas em idosos.

Dynamics of cell and tissue genesis in the male reproductive system of ticks (Acari: Ixodidae) Amblyomma cajennese (Fabricius, 1787) and Amblyomma aureolatum (Pallas, 1772): a comparative analysis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microsatellite analysis supports clonal propagation and reduced divergence of Trypanosoma vivax from asymptomatic to fatally infected livestock in South America compared to West Africa

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Blood Profile and Histology in Oral Infections Associated with Diabetes

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Healing Process of Autogenous Bone Graft in Spontaneously Hypertensive Rats Treated With Losartan: An Immunohistochemical and Histomorphometric Study

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Biostratigraphy and paleoecology of an unusual palynological record from the Aquidauana Formation, Late Pennsylvanian of Paraná Basin

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Apoptosis and reduced microvascular density of the lamina propria during tooth eruption in rats

During tooth eruption, structural and functional changes must occur in the lamina propria to establish the eruptive pathway. In this study, we evaluate the structural changes that occur during lamina propria degradation and focus these efforts on apoptosis and microvascular density. Fragments of maxilla containing the first molars from 9-, 11-, 13- and 16-day-old rats were fixed, decalcified and embedded in paraffin. The immunohistochemical detection of vascular endothelial growth factor (VEGF), caspase-3 and MAC387 (macrophage marker), and the TUNEL method were applied to the histological molar sections. The numerical density of TUNEL-positive cells and VEGF-positive blood vessel profiles were also obtained. Data were statistically evaluated using a one-way anova with the post-hoc Kruskal-Wallis or Tukey test and a significance level of P ≤ 0.05. Fragments of maxilla were embedded in Araldite for analysis under transmission electron microscopy (TEM). TUNEL-positive structures, fibroblasts with strongly basophilic nuclei and macrophages were observed in the lamina propria at all ages. Using TEM, we identified processes of fibroblasts or macrophages surrounding partially apoptotic cells. We found a high number of apoptotic cells in 11-, 13- and 16-day-old rats. We observed VEGF-positive blood vessel profiles at all ages, but a significant decrease in the numerical density was found in 13- and 16-day-old rats compared with 9-day-old rats. Therefore, the establishment of the eruptive pathway during the mucosal penetration stage depends on cell death by apoptosis, the phagocytic activity of fibroblasts and macrophages, and a decrease in the microvasculature due to vascular cell death. These data point to the importance of vascular rearrangement and vascular neoformation during tooth eruption and the development of oral mucosa.

Prevalence and geographical distribution of Toxoplasma gondii in dogs in the urban area of Botucatu, SP, Brazil

Toxoplasmosis is a worldwide zoonosis caused by Toxoplasma gondii, and can infect a wide variety of animals including humans. Domestic animals can be an important sentinel population for infection in the community. Occurrence of T. gondii infection was assessed in dogs in the urban area of Botucatu city, SP, Brazil. In the sample, 10% rate for error estimate, 95% confidence interval, and 5% significance level were established. Serum samples were collected from dogs during a rabies vaccination campaign, and were processed using modified agglutination test (MAT). Blood samples were collected from 670 dogs, with homogeneous distribution in five regions in the urban area, representing 3.74% of 17,910 animals vaccinated. In this sample, 17.3% (116/670) dogs (68 – 58.6% female and 48 – 41.4% male) were positive for T. gondii infection (p<0.03). Regarding age of the infected dogs 4.6% (4/88) were younger and 95.4% (84/88) were older than one year (p<0.01); the age of 28 positive animals were undetermined. The serum titers of anti-T. gondii antibodies were: 16 (69.8%; 81/116), 64 (13.8%; 16/116), 256 (15.5%; 18/116), and 1024 (0.9%; 1/116). Prevalence was distributed among the North 14.2% (19/134), South 18.0% (31/172), East 15.7% (19/121), West 21.6% (27/125), and Central 16.9% (20/118) regions of the municipality (p=0.5). In all these regions, females and dogs aged more than one year showed a higher occurrence of T. gondii infection (p<0.05).

An Ardent Flame: Witness to Distant Suffering, Human Rights and Unworthy Victims in the Coverage by The New York Times and Two Journals of the Religious Left of the 1980s Civil Wars in El Salvador and Nicaragua

Scholars have investigated witness to distant suffering (WTDS) almost entirely in visual media. This study examines it in print. This form of reporting will be examined in two publications of the religious left as contrasted with the New York Times. The thesis is that, more than any technology, WTDS consists of the journalist’s moral commitment and narrative skills and the audience’s analytical resources and trust. In the religious journals, liberation theology provides the moral commitment, the writers and editors the narrative skills and trust and the special vision of the newly empowered poor the analytical foundation. In bearing witness to those who have suffered state or guerilla terrorism in El Salvador and Nicaragua during the 1980s, we will investigate a distinction between “worthy” and “unworthy victims.” This last issue has a special ethical and political significance. Media witnessing to the suffering of strangers can help them become known, and so “worthy.” It can help them, and their plight and cause, become better recognized. This is the power of the media.

Anemia prevalence and its determinants in Brazilian institutionalized elderly

Objective: To estimate the prevalence of anemia and analyze the factors associated with anemia in elderly residents of long-term care institutions. Methods: This cross-sectional study was performed in male and female elderly volunteers selected in a two-stage random sampling from long-term care institutions in the city of Maringa, Brazil (2008). A diagnosis of anemia was based on the plasma hemoglobin concentration. The independent variables analyzed were gender, age, time of residence at an institution, body mass index, and serum iron and albumin concentrations. The association between anemia and the variables was assessed using the Poisson regression with robust variance in unadjusted and adjusted analyses, considering a complex sample and a significance level of 5%. Results: The sample included 124 adults older than 60 y residing in long-term care institutions (53.0% female). The prevalence of anemia was 29% and was not significantly associated with gender, serum iron concentration, time of residence at an institution, or body mass index. Conversely, hypoalbuminemia was considered a risk factor for anemia. Conclusion: There is a high prevalence of anemia in the institutionalized elderly and hypoalbuminemia is a factor associated with this outcome. Interventions are necessary to promote improvements in the health and welfare of this population. (C) 2012 Published by Elsevier Inc.