Human papillomavirus (HPV) infection is the most common sexually transmitted disease worldwide and there is a strong link between certain high-risk viral types and cervical carcinogenesis. Although there are several typing methods, it is still unclear which test is the best. This study compared the effectiveness of type-specific PCR (TS-PCR) and sequencing, with a focus on their clinical application. A total of 260 cervical samples from HPV-positive patients were tested for types 6, 11, 16, 18, 31, 33 and 35 using TS-PCR and sequencing. The genotype was identified in 36% of cases by TS-PCR and in 75% by sequencing. Sequencing was four times more likely to identify the viral type in positive samples than TS-PCR (p = 0.00). Despite being more effective for virus genotyping, sequencing was unable to identify viral types in multiple infections. Combining both techniques resulted in highly sensitive detection (87% of cases), showing that they are complementary methods. HPV genotyping is an important step in HPV management, helping to identify patients with a higher risk of developing cervical cancer and contributing to the development of type-specific vaccines.

The molecular basis of familial hypercholesterolemia (FH) in three families of Spanish descent from La Habana was investigated by the candidate gene approach. The Arg3500Gln mutation of apolipoprotein B-100 was not found. Identification of low density lipoprotein receptor (LDLR) gene haplotypes segregating with FH guided the characterisation of three point mutations by automated sequencing. One, a Val408-->Met missense mutation, a founder mutation in Afrikaner FH patients, was recurrent, being associated with a distinct DNA haplotype. The other two, Glu256-->Lys and Val776-->Met missense mutations, were novel and modified highly conserved residues. These mutations were absent in normolipidemic subjects and were associated in heterozygous carriers with twice the cholesterol levels observed in non-carriers. Noticeably, cardiovascular complications were rarely observed in older heterozygotes, even in those with the Afrikaner FH-2 mutation. These findings confirm the molecular heterogeneity of LDLR gene mutations causing FH and the variability of their expression across different populations.

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A, and 711+1G>T) in a sample of 111 newborn patients with cystic fibrosis diagnosed by the Cystic Fibrosis Neonatal Screening Program of Minas Gerais State. The mutations were tested by allele-specific oligonucleotide PCR with specially designed primers. An allele frequency of 48.2% was observed for the F508del mutation, and allele frequencies of 5.41, 4.50, 4.05, and 3.60% were found for the R1162X, G542X, 3120+1G>A, and G85E mutations, respectively. The genotypes obtained were in Hardy-Weinberg equilibrium. These data demonstrate that the 8-mutation panel studied here has extensive coverage (68%) for the cystic fibrosis mutations in Minas Gerais. These data improve our knowledge of cystic fibrosis in Brazil, particularly in this region. In addition, this investigation contributed to the establishment of a sensitive and population-specific mutation panel, which can be helpful for molecular diagnosis of cystic fibrosis.]]>

Objective: To assess the incidence of biotinidase deficiency among newborns and their clinical outcome up to one year of age in a large pilot screening study in Minas Gerais, Brazil.Methods: A prospective cohort study was conducted from September 2007 to June 2008 with heel-prick blood samples collected on filter paper for the purpose of newborn screening. A qualitative colorimetric test was used as the primary screening method. Colorimetricpositive cases were further tested with a serum confirmatory assay. Gene sequencing was performed for eight children suspected with biotinidase deficiency and for some of their parents. Positive cases were daily supplemented with oral biotin and were followed up for approximately six years.

The CIN incidence was low despite the high HPV prevalence. Being younger than 19 years at first sexual intercourse and not using antiretroviral medications were found to be the main risk factors for CIN.

RESUMOObjetivo: avaliar a associação entre a contagem de linfócitos T CD4+ e a gravidade da neoplasia intra-epitelial cervical em pacientes HIV positivas. Métodos: estudo transversal no qual foram incluídas 87 pacientes infectadas pelo HIV, confi rmado por testes sorológicos prévios. Todas eram portadoras do HPV cervical, diagnosticado por meio da reação em cadeia da polimerase. Foram realizados anamnese, exame físico e colposcopia de todas em pacientes. A biópsia do colo uterino foi realizada quando indicada pelo exame colposcópico. Os resultados histopatológicos foram classifi cados com neoplasia intra-epitelial de baixo grau (NIC I) ou de alto grau (NIC II e II). A associação entre a contagem de linfócitos T CD4+ e a gravidade da lesão foi verifi cada por meio da comparação de médias utilizando a análise da variância (ANOVA). Resultados: entre as 60 pacientes biopsiadas foram encontrados 24 casos (40,0%) com NIC I, oito (13,3%) NIC II, três (5%) NIC III, 14 (23,3%) pacientes somente com cervicite crônica e 11 (18,3%) apresentando efeito citopático produzido pelo HPV, mas sem perda da polaridade celular. Isso equivale a 35 mulheres com lesão intra-epitelial de baixo grau (NIC I + HPV) (58,3%) e 11 (18,3%) com lesão intra-epitelial de alto grau (NIC II + NIC III). A associação entre a média da contagem de linfócitos T CD4+ e a gravidade da lesão intra-epitelial cervical não foi signifi cativa (p=0,901). Conclusões: não houve associação entre a contagem de linfócitos T CD4+ e a gravidade da lesão intra-epitelial do colo uterino, diagnosticada pelo exame histopatológico. PALAVRAS-CHAVE:Neoplasia intra-epitelial cervical/patologia; Sondas DNA HPV; Neoplasias do colo do útero; Infecções por HIV; AIDS ABSTRACT Purpose: to evaluate association between CD4+ cell count and cervical intraepithelial lesion severity in HIV-infected women. Methods: cross-sectional study of 87 HIV-infected patients which were confi rmed by previous serologic examinations. All had cervical HPV diagnosed by polymerase chain reaction (PCR). All patients underwent anamnesis, physical examinations and colposcopy. Cervix biopsy was performed when indicated by colposcopical examination. Histopathological results followed Richart's classifi cation, adapted by Wright, and CD4+ cell count and cervical intraepithelial lesion severity association was analysed by comparison of means using analysis using analysis of variance (ANOVA).Results: among 60 biopsied women 24 were found (40.0%) with CIN I, eight (13.3%) with CIN II, three (5%) with CIN III, 14 (23.3%) with chronic cervicitis and 11 with cytopathic effect of HPV, without cell polarity loss. This corresponds to 35 (58.3%) women with intraepithelial lesion of low grade (CIN I + HPV) and 11 (18.3%) with intraepithelial lesion of high grade (CIN II + CIN III). There was no signifi cant association between CD4+ cell count mean and cervical intraepithelial lesion severity (p=0.901).Conclusions: there was no association between CD4+ cell count and cervical intraepithelial lesion severity diagnosed by hi...