X-linked insertion/deletion polymorphisms: forensic applications of a 33-markers panel

Insertion/deletion (INDEL) polymorphisms are diallelic markers with potential characteristics for use in forensics and biological anthropology, including: the simplicity of laboratory analysis, the possibility of genotyping many markers in a single PCR multiplex reaction, as well as analyzing markers with special inheritance types, such as those linked to the X chromosome (X-INDEL). In this work we developed a laboratory analysis methodology using a 33-INDEL marker panel for the X chromosome in a single PCR multiplex reaction, followed by a capillary electrophoresis run. We employed the panel to genotype a sample of 351 individuals of a mixed population from the Brazilian Amazon. The results demonstrate that the measurement of biostatistical parameters for forensic use in this population is compatible with prior estimates from other populations using current X-STR panels.     

Trends in social and demographic inequalities in the prevalence of chronic diseases in Brazil. PNAD: 2003- 2008

The aims of this study are: to evaluate the prevalence of chronic diseases in the Brazilian population comparing data of 2008 with those of 2003; to estimate the impact of chronic conditions on the use of health services and on the restriction of daily activities and to measure the differentials in the prevalence of specific diseases according to educational strata and the affiliation to a private health plan. Data were obtained from PNAD 2008 and 2003. The analysis included estimations of crude and adjusted prevalence ratios, using svy commands from Stata 11 software. The prevalence of at least one disease was higher in: the elderly, women, low schooling level, black or indigenous people, urban residents, migrants and people living in the south region of Brazil. The most frequent diseases were: hypertension, back and spinal cord disorders, arthritis and depression. Between 2003 and 2008, an increase in the prevalence of diabetes, hypertension, cancer and cirrhosis was observed, and there was a reduction in chronic kidney failure and tuberculosis. All the diseases analyzed, with the exception of cancer and tendinitis/tenossinovitis, revealed a higher prevalence in low educational level strata. The greatest social inequalities were in chronic kidney failure, cirrhosis, tuberculosis and arthritis/rheumatism.     

Safety and immunogenicity of a bivalent cytomegalovirus DNA vaccine in healthy adult subjects

BACKGROUND: VCL-CB01, a candidate cytomegalovirus (CMV) DNA vaccine that contains plasmids encoding CMV phosphoprotein 65 (pp65) and glycoprotein B (gB) to induce cellular and humoral immune responses and that is formulated with poloxamer CRL1005 and benzalkonium chloride to enhance immune responses, was evaluated in a phase 1 clinical trial. METHODS: VCL-CB01 was evaluated in 44 healthy adult subjects (22 CMV seronegative and 22 CMV seropositive) 18-43 years old. Thirty-two subjects received 1- or 5-mg doses of vaccine on a 0-, 2-, and 8-week schedule, and 12 subjects received 5-mg doses of vaccine on a 0-, 3-, 7-, and 28-day schedule. RESULTS: Overall, the vaccine was well tolerated, with no serious adverse events. Local reactions included mild to moderate injection site pain and tenderness, induration, and erythema. Systemic reactions included mild to moderate malaise and myalgia. All reactions resolved without sequelae. Through week 16 of the study, immunogenicity, as measured by enzyme-linked immunosorbant assay and/or ex vivo interferon (IFN)-gamma enzyme-linked immunospot assay, was documented in 45.5% of CMV-seronegative subjects and in 25.0% of CMV-seropositive subjects who received the full vaccine series, and 68.1% of CMV-seronegative subjects had memory IFN-gamma T cell responses at week 32. CONCLUSION: The safety and immunogenicity data from this trial support further evaluation of VCL-CB01.     

Antidepressants differentially modify the extinction of an aversive memory task in female rats

Treatment of major depression, posttraumatic stress disorder and other psychopathologies with antidepressants can be associated with improvement of the cognitive deficits related to these disorders. Although the mechanisms of these effects are not completely elucidated, alterations in the extinction of aversive memories are believed to play a role in these psychopathologies. We have recently verified that female rats present low levels of extinction when submitted to the plus-maze discriminative avoidance task. In the present study, female rats were treated long term with clinically used antidepressants (fluoxetine, nortriptyline or mirtazapine) and subjected to the plus-maze discriminative avoidance task to evaluate learning, memory, extinction and anxiety-related behaviors as well as behavioral despair in the forced swimming test. All groups learned the task and exhibited retrieval. Chronic treatment with fluoxetine (but not with the other antidepressants tested) increased extinction of the discriminative task. In the forced swimming test, the animals treated with fluoxetine and mirtazapine showed decreased immobility duration. In conclusion, fluoxetine potentiated extinction, while both fluoxetine and mirtazapine were effective in ameliorating depressive-like behavior in the forced swimming test, suggesting a possible dissociation between the effects on mood and the extinction of aversive memories in female rats.     

Attenuated nephritis in inducible nitric oxide synthase knockout C57BL/6 mice and pulmonary hemorrhage in CB17 SCID and recombination activating gene 1 knockout C57BL/6 mice infected with Leptospira interrogans

The aims of this study were to investigate the frequency of pulmonary hemorrhage (PH) in mice unable to produce functional B and T lymphocytes and to explore the effect of an inducible nitric oxide synthase gene (Inos) knockout (KO) on the frequency/severity of interstitial nephritis in vivo. We studied the outcome of infection by the virulent Leptospira interrogans serovar Copenhageni strain Cop. The animals used were Inos KO mice, recombination activating gene 1 (Rag1) KO mice, CB17 severe combined immunodeficiency (SCID) mice, and the respective wild-type (WT) C57BL/6 and BALB/c controls. The Inos KO and WT mice survived with no clinical symptoms of leptospirosis. The frequency and severity of nephritis was significantly lower in the Inos KO mice. All of the Rag1 KO and SCID animals died of acute leptospirosis, whereas all of the WT mice survived. PH was observed in 57 and 94% of Rag1 KO mice and in 83 and 100% of SCID mice, using inoculum doses of 10(7) and 10(6) leptospires, respectively. There was no evidence of PH in the WT controls. In conclusion, the loss of the Inos gene had a negligible effect on the outcome of leptospiral infection, although we observed a reduced susceptibility for interstitial nephritis in this group. Of note, the absence of functional B- and T-cell lymphocytes did not preclude the occurrence of PH. These data provide evidence that PH in leptospirosis may not be related only to autoimmune mechanisms.     

The anti-obesity agent Orlistat is associated to increase in colonic preneoplastic markers in rats treated with a chemical carcinogen

Orlistat is an anti-obesity agent that increases the fecal fat excretion, which promotes colon carcinogenesis. Therefore, the present study was designed to verify the effects of Orlistat on the formation of rat colonic aberrant crypt foci (ACF) and cell proliferation evaluated by the PCNA method. Male Wistar rats received either a standard diet or a high fat diet (HFD), supplemented or not with Orlistat (200 mg/kg chow) and two doses of the carcinogen dimethyl-hydrazine (25 mg/Kg). After 30 days, Orlistat was associated to a significant increase in the number of colonic ACFs and cell proliferation in DMH-treated animals, independently of the HFD.     

Factors associated with community-acquired pneumonia in hospitalised children and adolescents aged 6 months to 13 years old

According to the World Health Organisation, community-acquired pneumonia is the main cause of paediatric death, accounting for 20 % of deaths in children younger than 5 years old, and 90 % of these deaths occur in non-industrialised countries. This study has as objective to evaluate the influence of socio-economic, environmental and breastfeeding factors on the occurrence of pneumonia. An unmatched case–control study was conducted in children aged 6 months to 13 years old at a children’s hospital in Brazil. Multivariate analysis by logistic regression was performed to determine the variables used to predict pneumonia. A total of 252 children were selected. In the adjusted (by age) multivariate analysis, the following variables were associated with community-acquired pneumonia: (a) protective factors: breastfeeding >3 months, absence of other unrelated comorbidities, non-smoking mother, being the only child, child’s age >5 years and mother’s age >19 years old; (b) risk factors: maternal education <8 years and child’s birth order [≥second]. In the multivariate analysis, considering only children from 6 months to 5 years old, the following variables were associated with community-acquired pneumonia: (a) protective factors: breastfeeding >3 months, non-smoking mother and no smokers in the child’s bedroom; (b) risk factors: maternal education <8 years and prenatal complications. Conclusion: These findings contribute favourably to effectively minimising the risk factors related to the disease process and natural history of community-acquired pneumonia.