Unraveling the Significance of DGCR8 and miRNAs in Thyroid Carcinoma.

MicroRNAs (miRNAs) act as negative regulators for protein-coding gene expression impacting cell proliferation, differentiation, and survival. These miRNAs are frequently dysregulated in cancer and constitute classes of blood-based biomarkers useful for cancer detection and prognosis definition. In thyroid cancer (TC), the miRNA biogenesis pathway plays a pivotal role in thyroid gland formation, ensuring proper follicle development and hormone production. Several alterations in the miRNA biogenesis genes are reported as a causality for miRNA dysregulation. Mutations in microprocessor component genes are linked to an increased risk of developing TC; in particular, a recurrent mutation affecting DGCR8, the E518K. In this review, we explore these novel findings and resume the current state-of-the-art in miRNAs in thyroid carcinomas.

Generation of an Obese Diabetic Mouse Model upon Conditional Atrx Disruption.

Atrx loss was recently ascertained as insufficient to drive pancreatic neuroendocrine tumour (PanNET) formation in mice islets. We have identified a preponderant role of Atrx in the endocrine dysfunction in a Rip-Cre;AtrxKO genetically engineered mouse model (GEMM). To validate the impact of a different Cre-driver line, we used similar methodologies and characterised the Pdx1-Cre;AtrxKO (P.AtrxKO) GEMM to search for PanNET formation and endocrine fitness disruption for a period of up to 24 months. Male and female mice presented different phenotypes. Compared to P.AtrxWT, P.AtrxHOM males were heavier during the entire study period, hyperglycaemic between 3 and 12 mo., and glucose intolerant only from 6 mo.; in contrast, P.AtrxHOM females started exhibiting increased weight gains later (after 6 mo.), but diabetes or glucose intolerance was detected by 3 mo. Overall, all studied mice were overweight or obese from early ages, which challenged the histopathological evaluation of the pancreas and liver, especially after 12 mo. Noteworthily, losing Atrx predisposed mice to an increase in intrapancreatic fatty infiltration (FI), peripancreatic fat deposition, and macrovesicular steatosis. As expected, no animal developed PanNETs. An obese diabetic GEMM of disrupted Atrx is presented as potentially useful for metabolic studies and as a putative candidate for inserting additional tumourigenic genetic events.

DGCR8 Microprocessor Subunit Mutation and Expression Deregulation in Thyroid Lesions.

DGCR8 emerged recently as miRNAs biogenesis pathway protein with a highlighted role in thyroid disease. This study aimed to characterize this miRNA biogenesis component, in particular the p.(E518K) mutation and DGCR8 expression in a series of thyroid lesions. The series of thyroid lesions was genotyped for the c.1552G>A p.(E518K) mutation. When frozen tissue was available, DGCR8 mRNA expression was analysed by qPCR. Formalin-fixed paraffin-embedded tissues were studied for DGCR8 immunoexpression. We present for the first time the p.(E518K) mutation in a case of poorly differentiated thyroid carcinoma and present the deregulation of DGCR8 expression at mRNA level in follicular-patterned tumours. The obtained data solidify DGCR8 as another important player of miRNA-related gene mutations in thyroid tumorigenesis, particularly in follicular-patterned thyroid tumours.

A Rare Case of Spontaneous Intrauterine Skull Fracture.

Skull fractures are rare in newborns and normally caused by maternal abdominal trauma or complicated deliveries. However, in rare cases, these fractures are found in neonates born after an uneventful pregnancy and delivery. We report a case of a primigravida who underwent cesarean delivery due to failure of descent and malpositioning of the fetal head. After birth, a right temporoparietal fracture and congenital muscular torticollis were diagnosed. The newborn's blood tests showed hypocalcemia and relative hypoparathyroidism. Both mother and newborn presented low vitamin D levels. Serial imaging control showed gradual resolution of the lesions, with the newborn being discharged at the 10th day of life with vitamin D supplementation. This is an interesting case because the combination of three conditions - maternal and fetal hypovitaminosis D, congenital torticollis and malposition of the cephalic pole during labor - may have synergistically contributed to a spontaneous intrauterine skull fracture.

As fraturas do crânio são raras em recém-nascidos, sendo mais comummente causadas por trauma abdominal ou como complicação do parto. Contudo, em casos mais raros, estas fraturas são encontradas isoladamente, sem associação a intercorrências da gravidez ou do parto. Apresentamos o caso de uma primigesta submetida a cesariana por ausência de descida da apresentação e mau posicionamento da mesma no canal de parto. Após o nascimento, foi diagnosticada fratura temporoparietal direita e torcicolo congénito. Analiticamente, o recém-nascido apresentava hipocalcemia e hipoparatiroidismo relativo. A díade mãe - recém-nascido apresentaram hipovitaminose D. Estudos imagiológicos seriados demonstraram resolução gradual das lesões, possibilitando a alta do recém-nascido ao 10º dia de vida com suplementação de vitamina D. Este caso é interessante porque se conjugam três condições ­ hipovitaminose D materna e fetal, torcicolo congénito e má orientação do polo cefálico ­ que, conjuntamente, podem ter contribuído para a ocorrência de fratura craniana intrauterina espontânea.

Treatment of chronic HCV infection with DAAs in Rio de Janeiro/Brazil: SVR rates and baseline resistance analyses in NS5A and NS5B genes.

The selection of viral strains with resistance-associated substitutions at hepatitis C virus (HCV) NS5A and NS5B genes is considered one of the limiting factors for achieving sustained virologic response (SVR) to combination of direct-acting antivirals daclatasvir (DCV) and sofosbuvir (SOF). Since 2015, this interferon-free regimen has been available in Brazilian clinical routine for treating mono- and HCV/HIV-coinfected patients chronically infected with genotypes 1 and 3. Our aim was to assess SVR rate for Brazilian patients chronically infected with genotypes 1 and 3 after DCV/SOF therapy and the frequency of baseline RASs in HCV NS5A and NS5B genes. Serum samples were collected from 107 monoinfected patients and 25 HCV/HIV co-infected patients before antiviral therapy with DCV/SOF. Genetic diversity of NS5A and NS5B genes was assessed by direct nucleotide sequencing. Overall, SVR rate was 95.4% (126/132), and treatment failure occurred in five monoinfected and one HCV/HIV co-infected patient. NS5A RASs frequency was higher for HCV/HIV patients (28%) than monoinfected patients (16.8%). No difference was evidenced between mono- and HCV/HIV-coinfected groups (15% vs. 16%) regarding NS5B gene. Genotype (GT) 1b strains had significantly more baseline substitutions in NS5A (31.6%) than GT 1a and 3a. At least one primary NS5A RAS described in literature at loci 28, 30, 31 or 93 was identified in HCV GTs 1 strains for both groups. As for NS5B, RASs at positions 159 and 316 was observed only in GT 1b strains. This study highlighted that SVR rate in clinical routine in Brazil was similar to randomized clinical trials (89-98%). Our research provided genetic data about the circulation of resistant variants in Brazil. Despite its presence, most of identified baseline mutations did not negatively impact treatment outcome. Genetic diversity of circulating strains suggested that most of the Brazilian HCV chronic carriers are susceptible to new therapeutic regimens including recently approved DAAs.

Prevalence of baseline NS3 resistance-associated substitutions (RASs) on treatment with protease inhibitors in patients infected with HCV genotype 1.

BACKGROUND AND AIMS: Treatment for hepatitis C has evolved significantly with the licensing of direct-acting antiviral drugs (DAAs). However, one of the limiting factors of the effectiveness of antiviral therapy with protease inhibitors (PIs) is the emergence of resistance caused by point mutations. The aim of this study was to determine the prevalence of resistance-associated substitutions (RASs) in HCV NS3 gene in patients infected with genotype 1 before therapy with simeprevir. METHODS: A total of 73 serum samples from 15 treatment-experienced patients with boceprevir/telaprevir and 58 DAA-naïve patients were collected before therapy with DAAs simeprevir, daclatasvir and/or sofosbuvir. Presence of baseline resistance-associated substitutions (RAS) in the serine protease domain of HCV NS3 was analyzed by nucleotide sequencing followed by amino acid deduction. RESULTS: Overall RAS prevalence in this study was 13.7% (10/73). RAS prevalence for HCV subtype 1b was 17.4% (4/23) while for HCV subtype 1a was 12% (6/50). Primary mutations V36M/L and R155K were observed only in HCV subtype 1a, whereas T54S and Q80K were identified only in HCV subtype 1b. RAS V36M, which is related to reduction of susceptibility to second-generation PIs, was the most frequent in the study (6.9%; 5/73). CONCLUSIONS: Our results indicated that Brazilian isolates of HCV present a distinct pattern of RAS depending on the infecting viral subtype. In contrast to data from other countries, RAS Q80K prevalence in Brazil is low in HCV subtype 1a. This study improves the knowledge of genetic barrier for resistance to PIs involving RASs in chronically infected patients and its possible impact on an unsuccessful treatment outcome, information that might be crucial to upcoming decisions of incorporation of new DAAs in Brazilian guidelines of antiviral therapy against HCV infection.

Recombinant VP1 protein as a potential marker for the diagnosis of acute hepatitis A virus infection.

Since hepatitis A virus (HAV) production is time-consuming and expensive, the use of recombinant proteins may represent an alternative source of antigens for diagnostic purposes. The present study aimed to express, purify and evaluate the potential of recombinant VP1 protein (rVP1) as a marker for the diagnosis of acute HAV infection. The rVP1 was expressed and purified successfully from Escherichia coli. The purified rVP1 was used to establish an in-house enzyme-linked immunosorbent assay (ELISA-rVP1) for detection of IgM antibodies in sera from HAV-positive patients. For a cut-off point of 0.351, the sensitivity and specificity of ELISA-rVP1 were 100.0% and 95.0%, respectively. These results indicate that rVP1 may be a useful antigen for detection of IgM antibodies against HAV.

Tornar-se pai vivenciando a internação do filho em Unidade de Terapia Intensiva Neonatal / Becoming a father living the hospitalization of the child in the Neonatal Intensive Care Unit

Objetivo - Compreender a experiência de tornar-se pai vivenciando a hospitalização do filho recém-nascido em Unidade de Terapia Intensiva Neonatal (UTIN). Métodos - O referencial teórico do estudo foi o Interacionismo Simbólico e o referencial metodológico, o Interacionismo Interpretativo. A coleta de dados foi orientada pelo método biográfico. Participaram nove pais de recém-nascidos hospitalizados em UTIN. Resultados - A análise das narrativas permitiu a compreensão da experiência através das categorias: Aguardar a chegada do filho; Deparar-se com o imprevisto; Saber dos riscos do filho; Viver o medo da perda; Conviver com a angústia de ter o filho na UTI; Ter a rotina da vida alterada; Esperar ansiosamente por estar junto ao filho; Aprender com o sofrimento; Prosseguir movido pela fé. Conclusão - A construção da parentalidade na situação de ter um filho em UTIN se inicia durante a espera da criança e permanece por toda hospitalização, apesar do distanciamento físico. A enfermagem tem um papel fundamental como elemento facilitador, oferecendo suporte acolhedor na difícil trajetória de internação do filho.

Objective - To comprehend the experience of becoming a father living the hospitalization of the newborn in the Neonatal Intensive Care Unit (NICU). Methods - The theoretical framework was the Symbolic Interactionism and the methodological framework was the Interpretative Interactionism. The data collection was guided by the biographic method. Nine fathers of hospitalized newborn in the NICU participated in this study. Results - The narrative analysis allowed the understand of the process, represented by the following categories: Waiting the arrival of the child; To be faced with the unexpected; To know the risks of the child; Living the fear of loss; To live with the distress of having the sonin the NICU; To have the life routine altered; Waiting to be with the son anxiously; To learn from suffering; To proceed moved by faith. Conclusion - The construction of paternity in this situation begins during waiting of the child and stays during the entire hospitalization period, despite of the physical distancing. Nursing has a fundamental role as a facilitator element, offering welcoming support in the difficult course of the hospitalization of the newborn.

[Anogenital warts in children: the importance of a multidisciplinary approach]. / Verrugas anogenitais na criança: a importância da abordagem multidisciplinar.

INTRODUCTION: Anogenital warts are caused by human papillomavirus (HPV) infection. Its presence in children raises concern of a possible sexual abuse. A multidisciplinary team approach is essential to clarify the mode of infection's transmission. CASE-REPORT: Female child, three years of age referred to pediatrician for perianal warts. A history of similar lesions in another location in the child, parents or cohabiting was denied and possible sexual abuse refused. Gynecologic assessment of mother and daughter lead to identification of HPV type 6 at the child's lesions and negative studies results in the mother. Evaluation of the child in psychology showed no signs of possible abuse and social assessment concluded there was no context of risk. After two cycles of topical treatment with imiquimod 5%, complete regression of lesions was achieved, with no recurrence to date. DISCUSSION: No evidence of sexual abuse emerged from the set of clinical evaluations performed. The strategy of an extended follow-up is mandatory, and comes from the required prudence in this context, in order to ensure timely identification of risk situations which might have been previously unnoticed. CONCLUSION: Authors emphasize the dichotomy between a simple clinical diagnosis and the complex multidisciplinary approach which is crucial to elucidate a situation with potential socio-medico-legal implications.

Superação de dormência em sementes de Butia capitata / Overcoming dormancy in seeds of Butia capitata

Com o objetivo de identificar métodos eficientes para a superação da dormência de sementes de B. capitata (Mart.) Becc. semeadas in vitro e em germinador, foram conduzidos testes com escarificação mecânica em pré-semeadura, através da abertura parcial ou total da cavidade embrionária de sementes isoladas dos endocarpos. A abertura da cavidade embrionária acelerou significativamente a germinação, principalmente quando houve retirada total do opérculo da semente, permitindo a germinação de, em média, 90 por cento dos embriões, independentemente da procedência dos acessos. A dormência das sementes de B. capitata parece estar relacionada com a barreira mecânica imposta pelos tecidos da semente que dificultam o desenvolvimento do embrião, o que sugere dormência exógena mecânica.

Aiming to identify efficient conditions to break dormancy in Butia capitata (Mart.) Becc. seeds sown in vitro and in an incubator tests were conducted with mechanical scarification in pre-sowing, by partial or total opening of the seed embryonic cavity, isolated of the endocarps. The embryonic cavity opening accelerated germination significantly, especially when there was total removal of the seed cap, allowing germination on average 90 percent of embryos, regardless of the provenance of the accessions. The seed dormancy of B. capitata seems to be related to the mechanical barrier imposed by the seed tissues that hamper the embryo development, suggesting mechanical exogenous dormancy.

Isolation and culture of soybean (Glycine max L. Merrill) microspores and pollen grains

In the last three decades, research on soybean microspore embryogenesis was restricted to anther culture, which presents limitations such as the small number of responsive microspores and the high embryogenic potential of sporophytic tissues. Therefore, a sequence of studies was performed to establish appropriate conditions for the isolation and culture of soybean microspores and pollen grains as an alternative to anther culture. First, a pollen and microspore isolation technique was developed using floral buds from four soybean cultivars (Bragg, IAS 5, MG/BR-46 Conquista and BRSMT Uirapuru). This technique allowed the establishment of cultures with satisfactory density and characteristics. Subsequently, different culture conditions were tested. Although B5 and MS media have been currently recommended for soybean anther culture, the best result was obtained in PTA-15 modified medium, with the formation of enlarged microspores and 0.4 percent of multicellular pollen grains in the cultivar BRSMT Uirapuru.

Nas últimas três décadas, a pesquisa em embriogênese do micrósporo de soja restringiu-se ao cultivo in vitro de anteras, com inúmeras limitações, como o pequeno número de micrósporos responsivos e o alto potencial embriogênico dos tecidos esporofíticos. Por isso, foi executada uma seqüência de testes visando ao estabelecimento de condições adequadas para o isolamento e o cultivo in vitro de micrósporos e grãos de pólen, como um sistema alternativo ao cultivo de anteras. Inicialmente, uma técnica de isolamento foi desenvolvida usando botões florais de quatro cultivares de soja (Bragg, IAS 5, MG/BR-46 Conquista e BRSMT Uirapuru), a qual possibilitou o estabelecimento de cultivos com características e densidade satisfatórias. Posteriormente, diferentes condições de cultivo foram testadas. Apesar de os meios B5 e MS serem recomendados para o cultivo de anteras de soja, o melhor resultado foi obtido em meio PTA-15 modificado, como o aumento do tamanho dos micrósporos e a formação de 0,4 por cento de grãos de pólen multicelulares na cultivar BRSMT Uirapuru.