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.

Nucleotide and phylogenetic analysis of human papillomavirus types 6 and 11 isolated from recurrent respiratory papillomatosis in Brazil

There are few studies about the distribution of natural molecular variants of low-risk HPVs. Our aim was to evaluate the E6 early gene variability among HPV-6 and HPV-11 isolates detected in recurrent respiratory papillomatosis (RRP) samples obtained in a cohort of Brazilian patients. We also performed a phylogenetic analysis in order to compare nucleotide sequences identified in our study with previously reported isolates from different anatomic sites (laryngeal papillomas, genital warts, cervical cancer and anal swabs) obtained from other parts of the world to determine the phylogenetic relationships of variants detected in Brazil. The complete coding region of the E6 gene of 25 samples was cloned and sequenced: 18 isolates of HPV-6 (72%) and 7 isolates of HPV-11 (28%). A total of four different HPV-6 genomic variants and two HPV-11 genomic variants was identified. It was not possible to correlate specific variants with disease severity. Phylogenetic trees for both HPV types were constructed enclosing both E6 sequences detected in our study and formerly published sequences. In both phylogenetic trees, the sequences from Brazil did not group together. We could not establish a geographical association between HPV-6 or HPV-11 variants, unlike HPV-16 and HPV-18. © 2013 Elsevier B.V.

Musculoskeletal disorders (MSDs) in dancers and former dancers participating in the largest dance festival in the world

Objective: To verify the presence of musculoskeletal disorders (MSD) in dancers and former dancers participating in the 27th Joinville Dance FestivalMaterial and methods: The research was conducted at the 27th Joinville Dance Festival in Santa Catarina, Brazil, with 173 participants in the competition, where 139 were dancers and 34 were former dancers with at least ten years of practiceIn order to obtain information related to MSDs, dancers answered a specific questionnaireResults: The study group consisted of adults (28±9 years) with body mass index within the normal range (21.3±2.3) with high weekly training time (955.7±837.2) and featuring long-time experience in dancing (18±7years)The lumbar spine and knees were the most affected by MSDsThe pain of high intensity was the symptom that most often appeared in reports among survey participantsConclusion: It is important to have deeper knowledge of the prevalence of pain among professional dancers, the factors associated with pain conditions and its impact on everyday life in order to plan new forms of prevention and plan new forms of prevention and treatment within a multidisciplinary approach© 2012 Elsevier Masson SAS.

Qualidade de vida de crianças e adolescentes asmáticos: Sua relação com estratégias de enfrentamento materno

Objective: To evaluate the quality of life of asthmatic children and adolescents, its relation with sociodemographic and clinical variables, and maternal coping strategies. Methods: Cross-sectional study in which children and adolescents with asthma answered a quality of life questionnaire, and their mothers did the same with a coping scale. Results: Out of the 42 children and adolescents investigated, 74% were classified as having mild/severe persistent asthma; 19%, mild persistent asthma; and 7%, intermittent asthma. A total of 69% of the participants showed impaired quality of life with mean scores ranging between 4.7 and 3.5, with greater harm in the domain of symptoms (score=3.6). There was a significant association between maternal schooling and the general index of quality of life, whereas maternal coping strategies were not associated with the severity of asthma. A large number of strategies used by mothers to cope with their children's crises were related to the management of stressors or to religious practices, and the latter presented negative correlation with the children's quality of life general index, showing that mothers whose children had worse quality of life used more religious coping. Conclusions: Asthmatic children, particularly those with moderate/severe persistent asthma, showed significant alterations as to quality of life. The high percentage of mothers using religious strategies, particularly in face of more severe clinical conditions, seem to indicate that they feel powerless to act, thus requiring concrete and useful orientation to low income families.

Prevalência de sobrepeso/obesidade e nível econômico de escolares

Objective: To associate the nutritional status of elementary schoolchildren with their gender, age group, and economic status based on a cross-sectional study carried out at public schools in the municipality of Cruzeiro do Oeste, Southern Brazil. Methods: The study was performed in 2010 including 939 schoolchildren aged six to ten years-old. The economical status of the families was classified based on a socioeconomic ranking questionnaire (Brazilian Association of Survey Companies), and the body mass index was calculated. Data were analyzed by descriptive statistics, and differences between independent groups were verified by Mann-Whitney and Kruskal-Wallis tests, while possible associations were verified by the chi-square test, being significant p<0.05. Results: The prevalence of obesity in the sample was 8.0%, and overweight, 16.4%. No association was found between the socioeconomic and nutritional status or between the nutritional status and gender. Conclusions: A high prevalence of overweight and obesity was found in this study, indicating that children of both genders are affected regardless of their economical status and age.

CDKN2A promoter hypermethylation in astrocytomas is associated with age and sex

CDKN2A promoter hypermethylation has been widely related to many cancers. In astrocytomas, although CDKN2A (p16INK4A protein) is often inactivated, there are still some controversial issues regarding the mechanism by which this alteration occurs. Thus, we analyzed a series of astrocytomas to assess the association between CDKN2A expression and methylation of grade I-IV tumors (WHO) and clinicopathological parameters. DNA extracted from formalin-fixed paraffin-embedded material of 93 astrocytic tumors was available for CDKN2A promoter methylation analysis and p16INK4A expression by methylation-specific PCR and immunohistochemistry, respectively. A strong negative correlation between nuclear and cytoplasmic immunostaining and CDKN2A promoter methylation was found. Additionally, a significant negative correlation between CDKN2A promoter methylation and age was observed; also, female patients had statistically more CDKN2A methylated promoters (p=0.036) than men. In conclusion, CDKN2A inactivation by promoter methylation is a frequent event in astrocytomas and it is related to the age and sex of patients. © 2013 Surgical Associates Ltd.

Genomic Signatures Predict Poor Outcome in Undifferentiated Pleomorphic Sarcomas and Leiomyosarcomas

Undifferentiated high-grade pleomorphic sarcomas (UPSs) display aggressive clinical behavior and frequently develop local recurrence and distant metastasis. Because these sarcomas often share similar morphological patterns with other tumors, particularly leiomyosarcomas (LMSs), classification by exclusion is frequently used. In this study, array-based comparative genomic hybridization (array CGH) was used to analyze 20 UPS and 17 LMS samples from untreated patients. The LMS samples presented a lower frequency of genomic alterations compared with the UPS samples. The most frequently altered UPS regions involved gains at 20q13.33 and 7q22.1 and losses at 3p26.3. Gains at 8q24.3 and 19q13.12 and losses at 9p21.3 were frequently detected in the LMS samples. Of these regions, gains at 1q21.3, 11q12.2-q12.3, 16p11.2, and 19q13.12 were significantly associated with reduced overall survival times in LMS patients. A multivariate analysis revealed that gains at 1q21.3 were an independent prognostic marker of shorter survival times in LMS patients (HR = 13.76; P = 0.019). Although the copy number profiles of the UPS and LMS samples could not be distinguished using unsupervised hierarchical clustering analysis, one of the three clusters presented cases associated with poor prognostic outcome (P = 0.022). A relative copy number analysis for the ARNT, SLC27A3, and PBXIP1 genes was performed using quantitative real-time PCR in 11 LMS and 16 UPS samples. Gains at 1q21-q22 were observed in both tumor types, particularly in the UPS samples. These findings provide strong evidence for the existence of a genomic signature to predict poor outcome in a subset of UPS and LMS patients. © 2013 Silveira et al.

Prognostic factors associated with time to hCG remission in patients with low-risk postmolar gestational trophoblastic neoplasia

Objective The purpose of this study was to identify the clinical factors associated with time to hCG remission among women with low-risk postmolar GTN. Methods This study included a non-concurrent cohort of 328 patients diagnosed with low-risk postmolar GTN according to FIGO 2002 criteria. Associations of time to hCG remission with history of prior mole, molar histology, time to persistence, use of D&C at persistence, presence of metastatic disease, FIGO score, hCG values at persistence, type of first line therapy and use of multiagent chemotherapy were investigated with both univariate and multivariate analyses. Results Overall median time to remission was 46 days. Ten percent of the patients required multi-agent chemotherapy to achieve hCG remission. Multivariate analysis incorporating the variables significant on univariate analysis confirmed that complete molar histology (HR 1.45), metastatic disease (HR 1.66), use of multi-agent therapy (HR 2.00) and FIGO score (HR 1.82) were associated with longer time to remission. There was a linear relationship between FIGO score and time to hCG remission. Each 1-point increment in FIGO score was associated with an average 17-day increase in hCG remission time (95% CI: 12.5-21.6). Conclusions Complete mole histology prior to GTN, presence of metastatic disease, use of multi-agent therapy and higher FIGO score were independent factors associated with longer time to hCG remission in low-risk GTN. Identifying the prognostic factors associated with time to remission and effective counseling may help improve treatment planning and reduce anxiety in patients and their families. © 2013 Elsevier Inc. All rights reserved.

Intra-abdominal fat is related to metabolic syndrome and non-alcoholic fat liver disease in obese youth

Background: Previous studies have shown an association between adiposity, especially intra-abdominal adipose tissue, and hemodynamic/metabolic comorbidities in adults, however it is not clear in pediatric population. The aim of the study was to analyze the relationship between non-alcoholic fatty liver disease (NAFLD) and components of metabolic syndrome (MS) with values of intra-abdominal (IAAT) and subcutaneous (SCAT) adipose tissue in obese children and adolescents.Methods: Cross-sectional study. Subjects: 182 obese sedentary children and adolescents (aged 6 to 16 y), identified by the body mass index (BMI). Measurements: Body composition and trunk fat by dual-energy X-ray absorptiometry- DXA; lipid profile, blood pressure and pubertal stage were also assessed. NAFLD was classified as absent (0), mild (1), moderate (2) and severe (3), and intra-abdominal and subcutaneous abdominal fat thickness were identified by ultrasound. The MS was identified according to the cut offs proposed by World Health Organization adapted for children and adolescents. The chi-square test was used to compare categorical variables, and the binary logistic regression indicated the magnitude of the associations adjusted by potential cofounders (sex, age, maturation, NAFLD and HOMA-IR).Results: Higher quartile of SCAT was associated with elevated blood pressure (p = 0.015), but not associated with NAFLD (p = 0.665). Higher IAAT was positively associated with increased dyslipidemia (p = 0.001), MS (p = 0.013) and NAFLD (p = 0.005). Intermediate (p = 0.007) and highest (p = 0.001) quartile of IAAT were also associated with dyslipidemia, independently of age, sex, maturation, NAFLD and HOMA-IR (homeostatic model assessment-insulin resistance).Conclusion: Obese children and adolescents, with higher IAAT are more prone to develop MS and NAFLD than those with higher values of SCAT, independent of possible confounding variables. © 2013 Silveira et al.; licensee BioMed Central Ltd.

Sensorimotor Integration in Dyslexic Children under Different Sensory Stimulations

Dyslexic children, besides difficulties in mastering literacy, also show poor postural control that might be related to how sensory cues coming from different sensory channels are integrated into proper motor activity. Therefore, the aim of this study was to examine the relationship between sensory information and body sway, with visual and somatosensory information manipulated independent and concurrently, in dyslexic children. Thirty dyslexic and 30 non-dyslexic children were asked to stand as still as possible inside of a moving room either with eyes closed or open and either lightly touching a moveable surface or not for 60 seconds under five experimental conditions: (1) no vision and no touch; (2) moving room; (3) moving bar; (4) moving room and stationary touch; and (5) stationary room and moving bar. Body sway magnitude and the relationship between room/bar movement and body sway were examined. Results showed that dyslexic children swayed more than non-dyslexic children in all sensory condition. Moreover, in those trials with conflicting vision and touch manipulation, dyslexic children swayed less coherent with the stimulus manipulation compared to non-dyslexic children. Finally, dyslexic children showed higher body sway variability and applied higher force while touching the bar compared to non-dyslexic children. Based upon these results, we can suggest that dyslexic children are able to use visual and somatosensory information to control their posture and use the same underlying neural control processes as non-dyslexic children. However, dyslexic children show poorer performance and more variability while relating visual and somatosensory information and motor action even during a task that does not require an active cognitive and motor involvement. Further, in sensory conflict conditions, dyslexic children showed less coherent and more variable body sway. These results suggest that dyslexic children have difficulties in multisensory integration because they may suffer from integrating sensory cues coming from multiple sources. © 2013 Viana et al.

Dental fluorescence: Potential forensic use

In cases of identification of bones, skeletal segments or isolated bones, searching for biotypologic diagnostic data to estimate an individual's age enables comparing these data with those of missing individuals. Enamel, dentin and pulp undergo remarkable changes during an individual's life. The enamel becomes more mineralized, smoother and thinner, and deteriorates because of physiological and pathological factors. Dental pulp decreases in volume due to the deposition of secondary dentin; thus, the dentin becomes thicker with time. In natural teeth, the fluorescence phenomenon occurs in dentin and enamel and changes in those tissues may alter the expression of the natural tooth color. The aim of this study was to assess the correlation between age and teeth fluorescence for individuals from different age groups. The sample consisted of 66 randomly selected Brazilians of both genders aged 7-63 years old. They were divided into 6 groups: Group 1 - aged 7-12 years, Group 2 - aged 13-20 years, Group 3 - aged 21-30 years, Group 4 - aged 31-40 years, Group 5 - aged 41-50 years and Group 6 - aged between 51 and 63 years. Upper right or left central incisors were used for the study. Restored and aesthetic rehabilitated teeth were excluded from the sample. The measurement of tooth fluorescence was carried out via computer analysis of digital images using the software ScanWhite DMC/Darwin Systems - Brazil. It was observed that dental fluorescence decreases when comparing the age groups 21-30, 31-40, 41-50 and 51-63 years. The results also showed that there is a statistically significant difference between the groups 41-50 years and 21-30 years (p=. 0.005) and also among the group 51-63 years and all other groups (p< 0.005). It can be concluded that dental fluorescence is correlated with age and has a similar and stable behavior from 7 to 20 years of age. It reaches its maximum expected value at the age of 26.5 years and thereafter decreases. © 2013 Elsevier B.V.

Atrichia, abnormal EEG, epilepsy and mental retardation in two sisters

Two daughters of a nonconsanguineous couple are described. Both present mental retardation, epileptic seizures, congenital atrichia, histologically anomalous skin and abnormal EEG pattern. From a discussion of the literature on atrichia, the forms without involvement of teeth, nails and hidrosis, among which recessive inheritance prevails, are distinguished from each other. None of them coincide with the syndrome described here.

Alcohol levels in cerebrospinal fluid and blood samples from patients under pathological conditions

We measured alcohol levels by the Cordebard method in 148 CSF samples from individuals who had abstained from alcohol for at least 7 days prior to the beginning of the study. Each blood sample was accompanied by a CSF sample from the same patient. CSF samples found to be normal after analysis were used as controls. Mean alcohol concentration in blood did not differ significantly between the control group and the groups with altered CSF. The group with altered CSF had statistically higher alcohol levels in CSF than in blood. CSF lactate, glucose and protein levels were not correlated with alcohol level. The results suggest the presence of endogenous alcohol in the CSF, with levels increasing in the presence of pathological processes involving the nervous system.

Candida spp. biotypes in the oral cavity of school children from different socioeconomic categories in Piracicaba - SP, Brazil

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

Ergonomic research applied in the design of pre-school furniture: The Mobipresc 3.6

This research describes the application of a scientific and technological model of Ergonomics in the design of pre-school furniture. The constant presence of the desk in early education and its influence in the relationship between the user and his educational environment determined the necessity of this project. The pre-school desk was considered as a work station, where the joint aspects of education and child anthropometry substantiate the problem. The review of the Historical application of Ergonomics in the Design of children's products consolidated the importance of this report. The development of ergonomic research, characterised by investigations of the Brazilian child's Anthropometry Data and Biomechanical Features, resulted in dimensional parameters of the user and physical characteristics of the present furniture. These elements, together with a comprehension of activities and needs in the pre-school, were connected with aspects of bibliographical revision to result in a series of recomendations for design. Through the methods of Ergonomic Design, a new proposal for the pre-school desk was developed, denominated Mobipresc 3.6.