GBA1 variants in Brazilian Gaucher disease patients.

Gaucher disease (GD) is an autosomal recessive lysosomal disorder caused by pathogenic variants in GBA1 which result in the deficient activity of glucocerebrosidase (GCase). There are few data on the genetic characterization of Brazilian GD patients. This study aimed at characterizing the genotype of 72 unrelated Brazilian GD patients (type I = 63, type II = 4, type III = 5; male = 31). Forty patients were from South Brazil (SB), and 32 were from other regions of Brazil (Others). The exons and exon/intron junctions of GBA1 were analyzed by Sanger sequencing in 8 patients, or by massive parallel sequencing followed by Sanger of exons 9 and 10 in 64 patients. In total, 31 pathogenic variants were identified. The most frequent allele found was N370S (p.(Asn409Ser)) (41.0%), and the most frequent genotype was N370S/RecNciI p.[Asn409Ser];[Leu483Pro;Ala495Pro;Val499=](23.6%). Three variants (N370S - in exon 9, and RecNciI and L444P (p.(Leu483Pro), in exon 10) correspond to 76.3% of total alleles in SB and 59.4% in Others. Two novel variants were described: c.326del(p.(Gln109Argfs*9)) and c.690G>A (p.(?)). Although sequencing all the exons of GBA1 is the gold-standard method for the genetic analysis of GD patients, a step analysis can be proposed for Brazilian patients, starting with analysis of exons 9 and 10. The N370S allele is the most frequently associated with GD in Brazil.

Elevated holo-transcobalamin in Gaucher disease type II: A case report.

Gaucher disease (GD), one of the most common lysosomal disorders, is caused by deficiency of ß-glucocerebrosidase. Based on the presence and severity of neurological complications, GD is classified into types I, II (the most severe form), and III. Abnormalities in systemic markers of vitamin B12 (B12 ) metabolism have been reported in GD type I patients, suggesting a higher prevalence of B12 deficiency in these patients. A 2-month-old male with GD type II was admitted to the hospital presenting jaundice, hepatosplenomegaly, and ichthyosis. At admission, cholestasis and ascites, abnormal liver function enzymes, prolonged prothrombin time, and high levels of B12 were confirmed. Analysis of biomarkers of B12 status revealed elevated B12 and holo-transcobalamin (holo-TC) levels. The B12 profile found in our patient is the opposite to what is described for GD type I patients. Holo-TC may increase in inflammatory states or due to liver diseases. In GD, the accumulation of glucocerebroside may be a trigger that initiates a systemic inflammatory reaction, characterized by macrophage activation. We suggest higher levels of holo-TC could be associated with a more severe (neuronopathic) GD, and be a biomarker of GD type II.

Liver involvement in patients with Gaucher disease types I and III.

BACKGROUND & AIMS: Gaucher disease (GD) is a multisystemic disease. Liver involvement in GD is not well characterised and ranges from hepatomegaly to cirrhosis and hepatocellular carcinoma. We aim to describe, and assess the effect of treatment, on the hepatic phenotype of a cohort of patients with GD types I and II. METHODS: Retrospective study based on the review of the medical files of the Gaucher Reference Centre of the Hospital de Clínicas de Porto Alegre, Brazil. Data from all GD types I and III patients seen at the centre since 2003 were analysed. Variables were compared as pre- ("baseline") and post-treatment ("follow-up"). RESULTS: Forty-two patients (types I: 39, III: 3; female: 22; median age: 35 y; enzyme replacement therapy: 37; substrate reduction therapy: 2; non-treated: 3; median time on treatment-MTT: 124 months) were included. Liver enzyme abnormalities, hepatomegaly, and steatosis at baseline were seen in 19/28 (68%), 28/42 (67%), and 3/38 patients (8%), respectively; at follow-up, 21/38 (55%), 15/38 (39%) and 15/38 (39%). MRI iron quantification showed overload in 7/8 patients (treated: 7; MTT: 55 months), being severe in 2/7 (treated: 2/2; MTT: 44.5 months). Eight patients had liver biopsy (treated: 6; MTT: 58 months), with fibrosis in 3 (treated: 1; time on treatment: 108 months) and steatohepatitis in 2 (treated: 2; time on treatment: 69 and 185 months). One patient developed hepatocellular carcinoma. CONCLUSIONS: GD is a heterogeneous disease that causes different patterns of liver damage even during treatment. Although treatment improves the hepatocellular damage, it is associated with an increased rate of steatosis. This study highlights the importance of a follow-up of liver integrity in these patients.

Assessment of cellular cobalamin metabolism in Gaucher disease.

BACKGROUND: Gaucher disease (GD) is a lysosomal disorder caused by biallelic pathogenic mutations in the GBA1 gene that encodes beta-glucosidase (GCase), and more rarely, by a deficiency in the GCase activator, saposin C. Clinically, GD manifests with heterogeneous multiorgan involvement mainly affecting hematological, hepatic and neurological axes. This disorder is divided into three types, based on the absence (type I) or presence and severity (types II and III) of involvement of the central nervous system. At the cellular level, deficiency of GBA1 disturbs lysosomal storage with buildup of glucocerebroside. The consequences of disturbed lysosomal metabolism on biochemical pathways that require lysosomal processing are unknown. Abnormal systemic markers of cobalamin (Cbl, B12) metabolism have been reported in patients with GD, suggesting impairments in lysosomal handling of Cbl or in its downstream utilization events. METHODS: Cultured skin fibroblasts from control humans (n = 3), from patients with GD types I (n = 1), II (n = 1) and III (n = 1) and an asymptomatic carrier of GD were examined for their GCase enzymatic activity and lysosomal compartment intactness. Control human and GD fibroblasts were cultured in growth medium with and without 500 nM hydroxocobalamin supplementation. Cellular cobalamin status was examined via determination of metabolomic markers in cell lysate (intracellular) and conditioned culture medium (extracellular). The presence of transcobalamin (TC) in whole cell lysates was examined by Western blot. RESULTS: Cultured skin fibroblasts from GD patients exhibited reduced GCase activity compared to healthy individuals and an asymptomatic carrier of GD, demonstrating a preserved disease phenotype in this cell type. The concentrations of total homocysteine (tHcy), methylmalonic acid (MMA), cysteine (Cys) and methionine (Met) in GD cells were comparable to control levels, except in one patient with GD III. The response of these metabolomic markers to supplementation with hydroxocobalamin (HOCbl) yielded variable results. The content of transcobalamin in whole cell lysates was comparable in control human and GD patients. CONCLUSIONS: Our results indicate that cobalamin transport and cellular processing pathways are overall protected from lysosomal storage damage in GD fibroblasts. Extending these studies to hepatocytes, macrophages and plasma will shed light on cell- and compartment-specific vitamin B12 metabolism in Gaucher disease.

Innovating the fight against syphilis: SIM Project / Inovando a luta contra a sífilis: Projeto SIM

Introduction: Syphilis is a major public health problem. Its incidence has increased in Brazil, particularly in the Southern Region. New tools are available, and immediate action is necessary. Objective: To describe the pilot study of an investigation aimed to assess the prevalence of syphilis, hepatitis B and C, and HIV and evaluate three strategies for adherence to syphilis treatment. Methods: A spontaneous sample of participants was evaluated with a structured questionnaire and underwent rapid tests for syphilis, HIV, and hepatitis B and C after signing an informed consent form (ICF). Rapid tests reagent for syphilis were confirmed by quantitative venereal disease research laboratory (VDRL) and Treponema pallidum hemagglutination assay (TPHA). Participants with confirmation of syphilis were randomized into three groups for follow-up: telephone calls, SIM app, and usual care at the health unit. Results: During a two-day pilot, 68 participants were included. Fourteen (20.6%) had tests reagent for syphilis, 1 (1.5%) for hepatitis B, 3 (4.4%) for hepatitis C, and 1 (1.5%) for HIV. Eight (57.1%) of the initial 14 individuals with rapid tests reagent for syphilis agreed to participate in the study. Out of the 8 rapid tests for syphilis, 2 (25%) were confirmed as active syphilis (>1/8). Conclusion: The prevalence of active syphilis estimated in this population was 3.5%. The demand for tests was high. The COVID-19 epidemic had a negative impact on the development of the study, which is ready for implementation. Discussions on the role of such a testing unit and the coverage of the research project in a context that requires increasing COVID-19-focused testing are fundamental for the future development of the project.

Introdução: A sífilis é um importante problema de saúde pública. A incidência tem aumentado no Brasil, principalmente na Região Sul. Novas ferramentas estão disponíveis e uma ação imediata é necessária. Objetivo: Descrever o estudo piloto de uma pesquisa que avalia a prevalência de sífilis, hepatites B e C e HIV e três estratégias de aderência ao seguimento do tratamento. Métodos: Uma amostra espontânea de participantes foi avaliada com um questionário estruturado e testes rápidos para sífilis, HIV e hepatites B e C foram realizados após assinatura do Termo de Consentimento Livre e Esclarecido (TCLE). Os testes rápidos reagentes para sífilis foram confirmados por VDRL (venereal disease research laboratory) quantitativo e hemaglutinação para sífilis (Treponema pallidum hemagglutination assay ­ TPHA). Os participantes com confirmação de sífilis foram randomizados em três grupos para acompanhamento: ligações telefônicas, aplicativo do SIM e cuidados habituais na unidade de saúde. Resultados: Durante um piloto de dois dias, 68 participantes foram incluídos. Quatorze (20,6%) tiveram testes reagentes para sífilis, 1 (1,5%) para hepatite B, 3 (4,4%) para hepatite C e 1 (1,5%) para HIV. Oito (57,1%) dos 14 casos iniciais com teste rápido reagente para sífilis aceitaram participar do estudo. Dos 8 testes rápidos para sífilis, 2 (25%) foram confirmados como sífilis ativa (>1/8). Conclusão: A prevalência de sífilis ativa estimada nesta população foi de 3,5%. A demanda por exames foi alta. A epidemia de COVID-19 impactou negativamente o desenvolvimento do estudo, que está pronto para implementação. A discussão sobre o papel desta espécie de unidade de teste e a abrangência do projeto de pesquisa em um contexto que pede a expansão de testes focados na COVID-19 são fundamentais para o desenvolvimento futuro do projeto.

Investigation of paternity with alleged father deceased or missing: analysis of success at the end of the report.

In this work we present a retrospective study of 858 cases of paternity investigation performed in Rio Grande do Sul, Southern Brazil, from 2007 to 2012, where the alleged father was deceased or missing. These cases represent 3.3% (858/26187) of paternity tests performed in that period. Considering the analysis of 17 DNA short tandem repeat loci, we present here the proportion of cases with conclusive results according to the number of relatives of the unavailable alleged father investigated and their kinship. The results show 81.0% (695/858) of cases with conclusive results and their characteristics.