Papilomavírus humano e anormalidades cervicais em mulheres do sistema de saúde privado e público no Estado do Rio de Janeiro, Brasil

This article reports the HPV status and cervical cytological abnormalities in patients attended at public and private gynecological services from Rio de Janeiro State. It also comments the performance of each HPV DNA tests used. A set of 454 women from private health clinics was tested by routine Capture Hybrid II HPV DNA assay. Among these, 58.4% presented HPV and nearly 90% of them were infected by high risk HPV types. However, this group presented few premalignant cervical lesions and no invasive cervical cancer was registered. We also studied 220 women from low income class attended at public health system. They were HPV tested by polymerase chain reaction using My09/11 primers followed by HPV typing with E6 specific primers. The overall HPV prevalence was 77.3%. They also showed a high percentage of high squamous intraepithelial lesion-HSIL (26.3%), and invasive cervical carcinoma (16.3%). HPV infection was found in 93.1% and 94.4% of them, respectively. The mean ages in both groups were 31.5 and 38 years, respectively. In series 1, HPV prevalence declined with age, data consistent with viral transient infection. In series 2, HPV prevalence did not decline, independent of age interval, supporting not only the idea of viral persistence into this group, but also regional epidemiological variations in the same geographic area. Significant cytological differences were seen between both groups. Normal and benign cases were the most prevalent cytological findings in series 1 while pre-malignant lesions were the most common diagnosis in the series 2. HPV prevalence in normal cases were statistically higher than those from series 1 (p < 0.001), indicating a higher exposure to HPV infection. Women from both samples were referred for previous abnormal cytology. However, socio-demographic evidence shows that women from series 1 have access to treatment more easily and faster than women from series 2 before the development of pre-malignant lesions. These data provides baseline support for the role of social inequalities linked to high risk HPV infection leading to cervical cancer. Broadly screening programs and the development of safe and effective vaccines against HPV would diminish the toll of this disease that affect mainly poor women.Este artigo analisa a infecção por HPV e anormalidades citológicas cervicais encontradas em pacientes atendidas em serviços ginecológicos dos sistemas de saúde público e privado do estado do Rio de Janeiro. O trabalho também avalia os testes utilizados para detecção de DNA do HPV em cada população estudada. Um grupo de 454 mulheres oriundas de serviços da rede privada de saúde foi testado por Captura do Híbrido II. Destas, 58,4% apresentaram infecção por HPV e cerca de 90% delas estavam infectadas por HPV de alto risco. Este grupo, entretanto, apresentava poucos casos de lesões cervicais pré-malígnas e nenhum caso de câncer. Estudamos, também, 220 mulheres de baixo nível econômico atendidas no serviço de saúde pública que foram testadas para HPV pela reação da polimerase em cadeia utilizando-se os oligonucleotídeos My09/My11. A identificação dos tipos foi efetuada por amplificação com oligonucleotídeos específicos para a região E6 do genoma viral. A prevalência de HPV nesta população foi de 77.3%, observando-se uma alta porcentagem de casos de neoplasias intraepiteliais cervicais de alto grau (26,3%) e de carcinoma cervical invasivo (16,3%). A infecção por HPV foi achada em, respectivamente, 93,1% e 94,4% destes casos. A média de idade em ambos os grupos era de 31,5 e 38 anos, respectivamente. Na série 1, a prevalência da infecção por HPV decresce com a idade, enquanto na série 2 ela não desaparece, dando suporte não só à idéia de persistência viral neste grupo, mas também a variações epidemiológicas na mesma área geográfica. Diferenças significativas foram vistas nos dois grupos. Casos normais e benignos foram incidentes na série 1, enquanto as lesões malígnas predominaram na série 2. Ao contrário, casos normais infectados por HPV eram prevalentes na série 2 (p < 0.001), indicando maior exposição ao vírus. Embora as mulheres de ambos os grupos tenham sido incluídas no estudo por apresentarem citologia anormal, evidências sócio-demográficas demonstram que mulheres da série 1 tem acesso mais fácil e rápido ao tratamento do que as mulheres da série 2 antes que as lesões pré-malígnas se desenvolvam. Estes resultados fornecem dados sobre o papel das desigualdades sociais associadas à infecção por HPV de alto risco na progressão do câncer cervical. Programas de prevenção abrangentes e o desenvolvimento de vacinas eficazes e seguras contra o HPV poderiam reduzir o tributo desta doença que afeta principalmente mulheres pobres

Divergent Levels of Marker Chromosomes in an hiPSC-Based Model of Psychosis

Summary In the process of generating presumably clonal human induced pluripotent stem cells (hiPSCs) from two carriers of a complex structural rearrangement, each having a psychotic disorder, we also serendipitously generated isogenic non-carrier control hiPSCs, finding that the rearrangement occurs as an extrachromosomal marker (mar) element. All confirmed carrier hiPSCs and differentiated neural progenitor cell lines were found to be mosaic. We caution that mar elements may be difficult to functionally evaluate in hiPSC cultures using currently available methods, as it is difficult to distinguish cells with and without mar elements in live mosaic cultures

ARMC4 Mutations Cause Primary Ciliary Dyskinesia with Randomization of Left/Right Body Asymmetry

The motive forces for ciliary movement are generated by large multiprotein complexes referred to as outer dynein arms (ODAs), which are preassembled in the cytoplasm prior to transport to the ciliary axonemal compartment. In humans, defects in structural components, docking complexes, or cytoplasmic assembly factors can cause primary ciliary dyskinesia (PCD), a disorder characterized by chronic airway disease and defects in laterality. By using combined high resolution copy-number variant and mutation analysis, we identified ARMC4 mutations in twelve PCD individuals whose cells showed reduced numbers of ODAs and severely impaired ciliary beating. Transient suppression in zebrafish and analysis of an ENU mouse mutant confirmed in both model organisms that ARMC4 is critical for left-right patterning. We demonstrate that ARMC4 is an axonemal protein that is necessary for proper targeting and anchoring of ODAs

Redução de aminoácidos em polpas de bacuri (Platonia insignis Mart), cupuaçu (Theobroma grandiflorum Willd ex-Spreng Schum) e murici (Byrsonima crassifolia L.) processado (aquecido e alcalinizado) Amino acids reduction in processed (heated and alkalinized) pulps of bacuri (Platonia insignis Mart), cupuaçu (Theobroma grandiflorum Willd ex-Spreng Schum) and murici (Byrsonima crassifolia L.)

A literatura científica é pobre a respeito de frutas da Amazônia, como o murici, e suas características químicas devem ser estudadas. Por isso, esta pesquisa teve por proposta determinar o perfil aminoacídico das polpas de bacuri, cupuaçu e murici sob diferentes valores de pH (3,3, 5,8, 8,0 e 12,0), sem aquecimento ou com aquecimento por 12 horas/100 ºC com agitação e refluxo. Valores de pH, glicose, frutose e sacarose também foram determinados nas polpas sem aquecimento. Os nutrientes foram determinados por CLAE (Cromatografia Líquida de Alta Eficiência). As polpas de bacuri, cupuaçu e murici apresentaram valores de pH 3,2, 3,6 e 3,35, respectivamente. A sacarose foi, quantitativamente, o principal carboidrato nas polpas de cupuaçu (38,34%) e bacuri (36,93%), sendo que os teores de frutose e glicose foram similares, tanto na polpa de cupuaçu (8,93% e 9,03%) como na de bacuri (12,63% e 11,65%), respectivamente. Em contraste, a polpa de murici foi quase isenta de sacarose (0,57%), mas não de frutose (11,51%) ou glicose (11,39%). Nas polpas sem aquecimento, os principais aminoácidos foram: ácido glutâmico (46,6 mg/kg), ácido aspártico (28,8 mg/kg) e arginina (25,3 mg/kg) na polpa de bacuri; ácido aspártico (56,3 mg/kg), ácido glutâmico (44,0 mg/kg) e alanina (24,2 mg/kg) na polpa de cupuaçu; prolina (73,5 mg/kg), ácido glutâmico (23,7 mg/kg) e ácido aspártico (23,5 mg/kg) na polpa de murici. O aquecimento reduziu as concentrações de todos os aminoácidos nas 3 polpas. O meio fortemente alcalino (pH 12) produziu a maior degradação de aminoácidos. Lisina foi mais sensível ao aquecimento do que outros aminoácidos em pH 12.<br>Scientific literature presents few studies about fruits of the Amazonia, like murici, and yours chemical characteristics should be studied. Therefore, amino acid profiles of the bacuri, cupuaçu and murici pulps were determined under different values of pH (3.3, 5.8, 8.0 and 12.0) with heating (12 hours/100 ºC, with stirring and refluxing) or without heating. Glucose, fructose, sucrose and pH values also were obtained in the pulps without heating. All nutrients were analised by HPLC. The pHs were: 3.2, 3.6 and 3.35 in the bacuri, cupuaçu and murici pulps, respectively. Sucrose (38.34% and 36.93%) was the major carbohydrate while fructose (8.93% and 12.63%) and glucose (9.03% and 11.65%) shown similar percentages in the cupuaçu and bacuri pulps. Murici pulp was almost free of sucrose (0.57%), but not of fructose (11.51%) or glucose (11.39%). In the pulps without heating the major amino acids were: glutamic acid (46.6 mg/kg), aspartic acid (28.8 mg/kg) and arginine (25.3 mg/kg) in the bacuri pulp; aspartic acid (56.3 mg/kg), glutamic acid (44.0 mg/kg) and alanine (24.2 mg/kg) in the cupuaçu pulp; proline (73.5 mg/kg), glutamic acid (23.7 mg/kg) and aspartic acid (23.5 mg/kg) in the murici pulp. The heating of the 3 pulps decresead the concentration of all amino acids. The medium strongly alkaline (pH 12) produced more degradation of the amino acids than others pHs. Lysine was more sensible to the heating than others amino acids in pH 12

Absence of heterozygosity due to template switching during replicative rearrangements

Item does not contain fulltextWe investigated complex genomic rearrangements (CGRs) consisting of triplication copy-number variants (CNVs) that were accompanied by extended regions of copy-number-neutral absence of heterozygosity (AOH) in subjects with multiple congenital abnormalities. Molecular analyses provided observational evidence that in humans, post-zygotically generated CGRs can lead to regional uniparental disomy (UPD) due to template switches between homologs versus sister chromatids by using microhomology to prime DNA replication-a prediction of the replicative repair model, MMBIR. Our findings suggest that replication-based mechanisms might underlie the formation of diverse types of genomic alterations (CGRs and AOH) implicated in constitutional disorders

DVL3 Alleles Resulting in a-1 Frameshift of the Last Exon Mediate Autosomal-Dominant Robinow Syndrome

Robinow syndrome is a rare congenital disorder characterized by mesomelic limb shortening, genital hypoplasia, and distinctive facial features. Recent reports have identified, in individuals with dominant Robinow syndrome, a specific type of variant characterized by being uniformly located in the penultimate exon of DVL1 and resulting in a −1 frameshift allele with a premature termination codon that escapes nonsense-mediated decay. Here, we studied a cohort of individuals who had been clinically diagnosed with Robinow syndrome but who had not received a molecular diagnosis from variant studies of DVL1, WNT5A, and ROR2. Because of the uniform location of frameshift variants in DVL1-mediated Robinow syndrome and the functional redundancy of DVL1, DVL2, and DVL3, we elected to pursue direct Sanger sequencing of the penultimate exon of DVL1 and its paralogs DVL2 and DVL3 to search for potential disease-associated variants. Remarkably, targeted sequencing identified five unrelated individuals harboring heterozygous, de novo frameshift variants in DVL3, including two splice acceptor mutations and three 1 bp deletions. Similar to the variants observed in DVL1-mediated Robinow syndrome, all variants in DVL3 result in a −1 frameshift, indicating that these highly specific alterations might be a common cause of dominant Robinow syndrome. Here, we review the current knowledge of these peculiar variant alleles in DVL1- and DVL3-mediated Robinow syndrome and further elucidate the phenotypic features present in subjects with DVL1 and DVL3 frameshift mutations

DVL1 frameshift mutations clustering in the penultimate exon cause autosomal-dominant Robinow syndrome

Item does not contain fulltextRobinow syndrome is a genetically heterogeneous disorder characterized by mesomelic limb shortening, genital hypoplasia, and distinctive facial features and for which both autosomal-recessive and autosomal-dominant inheritance patterns have been described. Causative variants in the non-canonical signaling gene WNT5A underlie a subset of autosomal-dominant Robinow syndrome (DRS) cases, but most individuals with DRS remain without a molecular diagnosis. We performed whole-exome sequencing in four unrelated DRS-affected individuals without coding mutations in WNT5A and found heterozygous DVL1 exon 14 mutations in three of them. Targeted Sanger sequencing in additional subjects with DRS uncovered DVL1 exon 14 mutations in five individuals, including a pair of monozygotic twins. In total, six distinct frameshift mutations were found in eight subjects, and all were heterozygous truncating variants within the penultimate exon of DVL1. In five families in which samples from unaffected parents were available, the variants were demonstrated to represent de novo mutations. All variant alleles are predicted to result in a premature termination codon within the last exon, escape nonsense-mediated decay (NMD), and most likely generate a C-terminally truncated protein with a distinct -1 reading-frame terminus. Study of the transcripts extracted from affected subjects' leukocytes confirmed expression of both wild-type and variant alleles, supporting the hypothesis that mutant mRNA escapes NMD. Genomic variants identified in our study suggest that truncation of the C-terminal domain of DVL1, a protein hypothesized to have a downstream role in the Wnt-5a non-canonical pathway, is a common cause of DRS