Abstract Hepatic glycogen storage diseases (GSD) are characterized by recurrent episodes of hypoglycemia, and anemia has been recognized as a frequent complication of these disorders.This was a convenience cross-sectional study to evaluate hepcidin and IL-6 concentrations in patients with hepatic GSD and their association with anemia and other parameters of iron metabolism.Levels of hepcidin, IL-6, and markers of iron metabolism were measured in 32 patients receiving uncooked cornstarch therapy for GSD (GSD Ia= 18; Ib= 7; III= 3; IXa= 3; IXb= 1; median age 9.5 years). IL-6 concentrations were compared to those of 8 individuals heterozygous for GSD. Nine patients were anemic and five patients had hepatic adenomas. IL-6 levels were higher in patients than in heterozygotes. Eight patients had hyperferritinemia, and one had elevated transferrin saturation as well. Hepcidin correlated positively with ferritin levels. IL-6 correlated with hemoglobin, iron, transferrin, and transferrin saturation. There was no correlation between hepcidin and IL-6 levels. Patients with GSD Ib had the highest IL-6 levels.Anemia is a common finding in hepatic GSD, especially in GSD Ib, the type of GSD associated with the highest IL-6 levels. These findings suggest that inflammation is strongly associated with development of anemia in GSD Ib. hypoglycemia disordersThis disorders This crosssectional cross sectional IL6 IL 6 IL- metabolismLevels Levels IL6, 6, 3 Ia 18 7 III IXa IXb 1 95 9 5 9. years. years . years) adenomas heterozygotes hyperferritinemia well hemoglobin levelsAnemia Anemia

Abstract Mucolipidosis II and III (MLII and MLIII) are autosomal recessive diseases caused by pathogenic variants in GNPTAB and GNPTG genes that lead to defects in GlcNAc-1-phosphotransferase. This enzyme adds mannose 6-phosphate residues to lysosomal hydrolases, which allows enzymes to enter lysosomes. Defective GlcNAc-1-phosphotransferase causes substrate accumulation and inflammation. These diseases have no treatment, and we hypothesized that the use of substrate reduction therapy and immunomodulation may be beneficial at the cell level and as a future therapeutic approach. Fibroblasts from two patients with MLIII alpha/beta and 2 patients with MLIII gamma as well as from one healthy control were treated with 10 µM miglustat, 20 µM genistein, and 20 µM thalidomide independently. ELISA assay and confocal immunofluorescence microscopy were used to evaluate the presence of heparan sulfate (HS) and the impact on substrate accumulation. ELISA assay showed HS reduction in all patients with the different treatments used (p=0.05). HS reduction was also observed by immunofluorescence microscopy. Our study produced encouraging results, since the reduction in substrate accumulation, even partial, may offer benefits to the phenotype of patients with inborn errors of metabolism. MLII GlcNAc1phosphotransferase. GlcNAc1phosphotransferase GlcNAcphosphotransferase GlcNAc 1 phosphotransferase. phosphotransferase 6phosphate phosphate 6 hydrolases lysosomes inflammation treatment approach alphabeta alpha beta miglustat genistein independently (HS p=0.05. p005 p p=0.05 . 0 05 (p=0.05) results partial metabolism p00 p=0.0 (p=0.05 p0 p=0. (p=0.0 p=0 (p=0. p= (p=0 (p= (p

RESUMO Objetivo: Investigar o impacto da reabilitação pulmonar (RP) em desfechos funcionais e na qualidade de vida relacionada à saúde (QVRS) em pacientes com fibrose pulmonar idiopática (FPI) em lista de espera para transplante de pulmão e em tratamento com antifibróticos (AF). Métodos: Estudo observacional retrospectivo com pacientes consecutivos com FPI em tratamento com pirfenidona ou nintedanibe (grupo AF) submetidos a RP entre janeiro de 2018 e março de 2020. O grupo AF e o grupo controle (pacientes com FPI que não estavam em tratamento com AF) participaram de um programa de RP com 36 sessões ao longo de 12 semanas. Após o término do programa, os participantes foram avaliados quanto à distância percorrida no teste de caminhada de seis minutos (DTC6) e à QVRS. A DTC6 e a QVRS pré e pós-RP foram comparadas intra e intergrupos. Resultados: Não houve diferença significativa entre os grupos AF e controle quanto às características basais, incluindo idade, limitação do fluxo aéreo, comorbidades e necessidade de oxigênio. O grupo AF apresentou um aumento significativo da DTC6 após 12 semanas de RP (tamanho do efeito: 0,77; p < 0,05); esse aumento também foi significativo na comparação intergrupos (tamanho do efeito: 0,55; p < 0,05). O grupo AF apresentou melhora significativa no componente físico da QVRS após 12 semanas (tamanho do efeito: 0,30; p < 0,05). Conclusões: Em pacientes com FPI submetidos a RP, a melhora na DTC6 e no componente físico da QVRS parece ser maior naqueles que estejam recebendo tratamento com AF do que naqueles que não o estejam. Objetivo (RP (QVRS (FPI AF. . (AF) Métodos 201 2020 3 1 DTC (DTC6 pósRP pós Resultados basais idade aéreo oxigênio tamanho efeito 0,77 077 0 77 0,05 005 05 0,05) 0,55 055 55 0,05. 0,30 030 30 Conclusões (AF 20 202 (DTC 0,7 07 7 0,0 00 0,5 5 0,3 03 2 0,

ABSTRACT Objective: To investigate the impact of pulmonary rehabilitation (PR) on functional outcomes and health-related quality of life (HRQoL) in idiopathic pulmonary fibrosis (IPF) patients placed on a lung transplant waitlist and receiving antifibrotic therapy (AFT). Methods: This was a retrospective observational study of consecutive IPF patients receiving AFT with either pirfenidone or nintedanib (the AFT group) and undergoing PR between January of 2018 and March of 2020. The AFT group and the control group (i.e., IPF patients not receiving AFT) participated in a 12-week PR program consisting of 36 sessions. After having completed the program, the study participants were evaluated for the six-minute walk distance (6MWD) and HRQoL. Pre- and post-PR 6MWD and HRQoL were compared within groups and between groups. Results: There was no significant difference between the AFT and control groups regarding baseline characteristics, including age, airflow limitation, comorbidities, and oxygen requirement. The AFT group had a significant increase in the 6MWD after 12 weeks of PR (effect size, 0.77; p < 0.05), this increase being significant in the between-group comparison as well (effect size, 0.55; p < 0.05). The AFT group showed a significant improvement in the physical component of HRQoL at 12 weeks (effect size, 0.30; p < 0.05). Conclusions: Among IPF patients undergoing PR, those receiving AFT appear to have greater improvements in the 6MWD and the physical component of HRQoL than do those not receiving AFT. Objective (PR healthrelated health related (HRQoL (IPF . (AFT) Methods 201 2020 i.e., ie i e (i.e. 12week week 3 sessions sixminute six minute MWD (6MWD Pre postPR post Results characteristics age limitation comorbidities requirement 1 effect size 0.77 077 0 77 0.05, 005 0.05 , 05 0.05) betweengroup 0.55 055 55 0.05. 0.30 030 30 Conclusions (AFT 20 202 i.e. (i.e 0.7 07 7 00 0.0 0.5 5 0.3 03 2 i.e 0.

Abstract Hyperhomocysteinemia and vitamin B12 deficiency have been reported in patients with phenylketonuria. In this study, total homocysteine (tHcy) and methylmalonic acid (MMA) levels were analyzed in samples from 25 phenylketonuria (PKU) patients. Comparisons were made between pre- and post-treatment values (n= 3); on treatment values, between periods with high and normal/low phenylalanine (Phe) levels (n= 20); and in women before, during and after pregnancy (n= 3). THcy levels decreased after treating PKU with metabolic formula (p=0.014). Except for a pregnant woman before pregnancy, none of the patients had tHcy values above the normal range. In fact, tHcy was < 5 μmol/L in 34% of the samples. We observed a decrease in Phe, tHcy, and tyrosine levels during pregnancy. MMA levels did not differ significantly, with values remaining in the normal range. These data indicate that there was no B12 deficiency in patients who adhere to the diet. In conclusion, in PKU patients treated with metabolic formula, tHcy is frequently not elevated, remaining even in the lower normal range in some patients. Thus, clinical follow-up and adherence to dietary treatment are crucial to prevent B12 deficiency. B B1 study (tHcy (MMA 2 (PKU pre posttreatment post n= n (n 3 3) normallow low Phe (Phe 20 20) 3. . p=0.014. p0014 p p=0.014 0 014 (p=0.014) fact μmolL μmol L 34 significantly diet conclusion elevated Thus followup follow up p001 p=0.01 01 (p=0.014 p00 p=0.0 (p=0.01 p0 p=0. (p=0.0 p=0 (p=0. p= (p=0 (p= (p

Abstract Reduced biotinidase activity is associated with a spectrum of deficiency ranging from total deficiency to heterozygous levels, a finding that is not always explained by the pathogenic variants observed in the BTD gene. The investigation of miRNAs, regulatory elements and variants in the 3’UTR region may present relevance in understanding the genotype-phenotype association. The aims of the study were to characterize the regulatory elements of the 3’UTR of the BTD gene and identify variants and miRNAs which may explain the discrepancies observed between genotype and biochemical phenotype. We evaluated 92 individuals with reduced biotinidase activity (level of heterozygotes = 33, borderline = 35, partial DB = 20 or total DB= 4) with previously determined BTD genotype. The 3’UTR of the BTD gene was Sanger sequenced. In silico analysis was performed to identify miRNAs and regulatory elements. No variants were found in the 3’UTR. We found 97 possible miRNAs associated with the BTD gene, 49 predicted miRNAs involved in the alanine, biotin, citrate and pyruvate metabolic pathways and 5 genes involved in biotin metabolism. Six AU-rich elements were found. Our data suggest variants in the 3'UTR of BTD do not explain the genotype-phenotype discrepancies found in Brazilian individuals with reduced biotinidase.

Abstract GlcNAc-1-phosphotransferase is a hexameric complex formed by subunits α, β, and γ, where the first two are encoded by the GNPTAB gene and the third by the GNPTG gene. Pathogenic variants identified in the GNPTAB gene cause the diseases Mucolipidosis II and III alpha/beta, which are severe and characterized by an overflow of lysosomal hydrolases into the extracellular environment, and their absence in lysosomal compartments causes an accumulation of non-degraded macromolecules. Methodology: a retrospective study that included 32 unrelated Brazilian patients with a clinical and genetic diagnosis of Mucolipidosis II/III alpha/beta. The regional frequency of the altered alleles was determined. Results: The patients were from all regions of Brazil. The most prevalent variants were c.3503_3504del, associated with the severe form of the disease, and c.1208T>C, associated with the milder form. Variant c.3503_3504del is the most frequently found in the Midwest, Northeast, and Southeast regions of Brazil. In the South, 42.8% of the alleles present the c.1196C>T variant. Conclusions: From the perspective of all patients diagnosed with Mucolipidosis II/III in Brazil, it is possible to conclude that different regions present allelic frequencies of specific pathogenic variants, which can be explained by the occurrence of a founding effect or high inbreeding rates.

Abstract Fructose-1,6-bisphosphatase (FBPase) deficiency is a rare inborn error of fructose metabolism caused by pathogenic variants in the FBP1 gene. As gluconeogenesis is affected, catabolic episodes can induce ketotic hypoglycemia in patients. FBP1 analysis is the most commonly used approach for the diagnosis of this disorder. Herein, a Brazilian patient is reported. The proband, a girl born to a consanguineous couple, presented with severe hypoglycemia crisis in the neonatal period. At the age 17 months, presented a new crisis accompanied by metabolic acidosis associated with a feverish episode. Genetic analysis was performed by next-generation sequencing (NGS), identifying the NM_000507.3:c.611_614del variant in homozygosis in the FBP1 gene. In silico analysis and 3D modeling were performed, suggesting that this variant is associated with a loss of sites for substrate and Mg2+ binding and for posttranslational modifications of FBPase. The c.611_614del variant is located in a repetitive region of the FBP1 gene that appears to be a hotspot for mutational events. This frameshift creates a premature termination codon in the last coding exon which escapes the nonsense-mediated decay mechanism, according to in silico analysis. This variant results in an intrinsically disordered protein with loss of substrate recognition and post-translational modification sites.

Abstract Citrullinemia type 1 (CTLNI), long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency (LCHADD), and mut0 methylmalonic acidemia (mut0 MMA) are inborn errors of metabolism (IEMs) associated with sudden unexpected death in infancy (SUDI). Its most common pathogenic variants are: c.1168G>A (CTLNI, ASS1 gene), c.1528G>C (LCHADD, HADHA gene), c.655A>T and c.1106G>A (mut0 MMA, MUT gene). Considering the absence of estimates regarding the incidence of these diseases in Brazil, this study sought to investigate the prevalence of its main pathogenic variants in a healthy population in the southern region of the country. A total of 1,000 healthy subjects from Rio Grande do Sul were included. Genotyping was performed by real-time PCR. Individuals found to be heterozygous for c.1528G>C underwent further acylcarnitine profile analysis by tandem mass spectrophotometry. Allele and genotype frequencies were calculated considering Hardy-Weinberg equilibrium. The c.1528G>C variant was detected in heterozygosity in two subjects (carrier frequency = 1:500; allele frequency = 0.001; minimum prevalence of LCHADD = 1: 1,000,000), whose acylcarnitine profiles were normal. Variants c.1168G>A, c.655A>T, and c.1106G>A were not identified. These results denote the rarity of these IEMs in Southern Brazil, highlighting the need to expand the investigation of IEMs in relation to infant morbidity and mortality within the country.

Abstract Mucolipidosis II and III (ML II and III) alpha/beta and ML III gamma are lysosomal diseases caused by GlcNAc-1-phosphotransferase deficiency. Previous data indicate that MLII patients have functionally impaired immune system that contributes to predisposition to infections.We evaluated the immunological phenotype of three Brazilian patients with ML III gamma. Our data suggest that the residual activity of GlcNAc-1-phosphotransferase in patients with ML III gamma is enough to allow the targeting of the lysosomal enzymes required for B-cell functions maintenance.

Resumo A sífilis gestacional é uma doença prevalente no Brasil e em outros países de baixa e média renda. A dessensibilização à penicilina é recomendada para mulheres grávidas com sífilis que são alérgicas a β-lactâmicos. Este é um estudo descritivo que utiliza registros médicos de 2011 a 2015 de um hospital público materno-fetal do Sul do Brasil com média de 3.600 partos anuais. Foram incluídos todos os casos de gestantes com sífilis e diagnóstico presuntivo de alergia a β-lactâmicos durante o período de estudo. As pacientes encaminhadas para dessensibilização originaram-se do serviço pré-natal hospitalar internamente, bem como dos serviços municipais e estaduais de atendimento pré-natal. A dessensibilização oral foi realizada na sala de emergência obstétrica, e a unidade de terapia intensiva estava disponível em todos os momentos para o atendimento de possíveis intercorrências. Dez pacientes foram submetidas à dessensibilização durante o período estudado. História pessoal de urticária foi a reação mais comum que exigiu dessensibilização à penicilina. Todas as pacientes toleraram bem o procedimento, e não mostraram reações adversas. Relatamos no presente manuscrito um programa bem-sucedido de dessensibilização oral à penicilina. Nenhuma das pacientes apresentou reações adversas ou complicações, corroborando a viabilidade e segurança do protocolo de dessensibilização. A administração oral de penicilina tem baixo custo, e otimiza o uso de tempo e recursos para o tratamento adequado de sífilis gestacional no cenário apresentado.

Abstract Gestational syphilis is a prevalent disease in Brazil and other low and medium income countries. Desensitization to penicillin is recommended for pregnant women with syphilis who are allergic to β-lactams. This is a descriptive study utilizing outpatient medical records from 2011 to 2015 from a mother and child hospital that is part of the national healthcare system in the South of Brazil, which performs an average of 3,600 birth assistances per year. All cases of pregnant women with syphilis and presumptive diagnosis of β-lactam allergy during the study period were included. The patients referred for desensitization originated from the hospital prenatal care service, as well as from municipal/state antenatal care services. Oral desensitization was performed in the obstetric emergency room, and adult and pediatric intensive care units were available at all times. Ten patients underwent desensitization during the period of study. Personal history of urticaria was the most common reaction that demanded desensitization. All patients tolerated the procedure well, and showed no adverse reactions.We report a successful program of oral desensitization. None of the patients presented adverse reactions or complications, a fact that corroborates the feasibility and safety of the desensitization protocol. Oral administration of penicillin comes at a low cost, and optimizes the use of time and resources.

Abstract This study described a broad clinical characterization of classical homocystinuria (HCU) in Brazil. This was a cross-sectional, observational study including clinical and biochemical data from 72 patients (60 families) from Brazil (South, n = 13; Southeast, n = 37; Northeast, n = 8; North, n = 1; and Midwest, n = 1). Parental consanguinity was reported in 42% of families. Ocular manifestations were the earliest detected symptom (53% of cases), the main reason for diagnostic suspicion (63% of cases), and the most prevalent manifestation at diagnosis (67% of cases). Pyridoxine responsiveness was observed in 14% of patients. Only 22% of nonresponsive patients on treatment had total homocysteine levels <100 mmol/L. Most commonly used treatment strategies were pyridoxine (93% of patients), folic acid (90%), betaine (74%), vitamin B12 (27%), and low-methionine diet + metabolic formula (17%). Most patients diagnosed with HCU in Brazil are late diagnosed, express a severe phenotype, and poor metabolic control. Milder forms of HCU are likely underrepresented due to underdiagnosis.

Com o avanço e a popularização de técnicas moleculares, a identificação de mutações genéticas que causam doenças aumentou drasticamente. Deste modo, o número de laboratórios disponíveis para investigar uma determinada doença e, consequentemente, o número de diagnósticos, aumentou ao longo do tempo. Embora seja necessário identificar mutações e fornecer diagnósticos, é crucial também desenvolver abordagens terapêuticas específicas baseadas nestas informações. Esta revisão tem como objetivo destacar os avanços recentes em terapias mutação-alvo com produtos químicos que mitiguem a patologia mutacional ao nível molecular, para doenças que, para a maior parte, não têm tratamento eficaz. Atualmente, existem várias estratégias sendo utilizadas para corrigir diferentes tipos de mutações, incluindo as seguintes: a identificação e caracterização de compostos readthroughtraducionais; redirecionamento de splicingmediado por oligonucleotídeos de sentido contrário; reparo por mismatch; e salto de exon. Estas terapias e outras abordagens são revistas neste artigo.

With the advance and popularization of molecular techniques, the identification of genetic mutations that cause diseases has increased dramatically. Thus, the number of laboratories available to investigate a given disorder and the number of subsequent diagnosis have increased over time. Although it is necessary to identify mutations and provide diagnosis, it is also critical to develop specific therapeutic approaches based on this information. This review aims to highlight recent advances in mutation-targeted therapies with chemicals that mitigate mutational pathology at the molecular level, for disorders that, for the most part, have no effective treatment. Currently, there are several strategies being used to correct different types of mutations, including the following: the identification and characterization of translational readthrough compounds; antisense oligonucleotide-mediated splicing redirection; mismatch repair; and exon skipping. These therapies and other approaches are reviewed in this paper.

Fundamentando-se na perspectiva histórico-cultural do desenvolvimento humano, o objetivo deste estudo foi compreender as concepções das crianças acerca da escola de educação infantil. A amostra foi composta por cinco díades de crianças de cinco anos de idade que frequentavam a pré-escola. Cenas e instruções organizadas em três sessões de brincar foram utilizadas para eliciar episódios compatíveis com os objetivos do estudo. Os dados foram submetidos à análise de conteúdo temática. Os resultados mostraram que as crianças destacaram o brincar, as atividades, o espaço físico, os professores e as regras em suas concepções sobre a escola. Esses achados indicam a importância de se pensar quais espaços e oportunidades para brincadeiras estão sendo disponibilizados nas escolas de educação infantil.

Based on the cultural historical perspective of human development, this investigation aimed at understanding children's concepts about preschool. The sample was composed of five 5-year-old dyads enrolled in preschool. Scenes and instructions organized along three play sessions were used to elicit episodes compatible with the questions of the study. Thematic content analysis was used to analyze the data. The results showed that play, school activities, characteristics of the physical environment, teachers and school rules were highlighted by children regarding their conceptions about school. These findings show the importance of thinking which spaces and opportunities preschools are actually providing for children.

Pesquisas proporcionaram evidências importantes sobre a relação entre estratégias específicas de estudo e um alto desempenho escolar. O objetivo desta investigação é elaborar e avaliar um curso de capacitação para professores de ensino fundamental, com a finalidade de desenvolver as habilidades cognitivas, metacognitivas e motivacionais para a aprendizagem em seus alunos. Utilizou-se um delineamento pré e pós-teste. Participaram 10 professores de 5ª série e 54 alunos. Os instrumentos utilizados junto aos alunos foram o Raven, a Escala de Avaliação para Motivação para Aprender, a Escala de Estratégias de Aprendizagem e o Teste de Desempenho Escolar. Para os professores, foram utilizados o Questionário Metacognitivo e um questionário de autoavaliação. Nas análises intra e entre grupos, os resultados mostraram o efeito tempo para a maioria das variáveis e o efeito grupo apenas para a variável estratégias de aprendizagem. Conclui-se que os professores podem atuar explicitamente como mediadores no desenvolvimento das habilidades cognitivas e metacognitivas de aprendizagem.

Researches brought important evidences on the relation between specific strategies of study and elementary school performance. The objective of this investigation is to elaborate and evaluate a qualification course for elementary school teachers, aiming at developing their students learning of cognitive, metacognitive and motivational abilities. A pre-posttest design was used. Ten 5th grade teachers and 54 students participated in the study. The instruments used with the students were the Raven, the Scale for the Evaluation of Motivation to Learn, the Strategies for Learning Scale and the School Performance Test. For the teachers, the Metacognitive Questionnaire and a self-evaluation questionnaire were used. Results of the within and between group analyses showed a time effect for most variables and a group effect only for the strategies of learning variable. It is possible to conclude that teachers may explicitly act as mediators in the development of cognitive abilities of learning.

Infantile GM1 gangliosidosis is caused by the absence or reduction of lysosomal beta-galactosidase activity. Studies conducted in Brazil have indicated that it is one of the most frequent lysosomal storage disorders in the southern part of the country. To assess the incidence of this disorder, 390 blood donors were tested for the presence of two common mutations (1622-1627insG and R59H) in the GLB1 gene. Another group, consisting of 26 GM1 patients, and the blood donors were tested for the presence of two polymorphisms (R521C and S532G), in an attempt to elucidate whether there is a founder effect. The frequencies of the R59H and 1622-1627insG mutations among the GM1 patients studied were 19.2% and 38.5%, respectively. The frequency of polymorphism S532G was 16.7%, whereas R521C was not found in the patients. The overall frequency of either R59H or 1622-1627insG was 57.7% of the disease-causing alleles. This epidemiological study suggested a carrier frequency of 1:58. Seven different haplotypes were found. The 1622-1627insG mutation was not found to be linked to any polymorphism, whereas linkage disequilibrium was found for haplotype 2 (R59H, S532G) (p < 0.001). These data confirm the high incidence of GM1 gangliosidosis and the high frequency of two common mutations in southern Brazil.