Two Conserved Cysteine Triads in Human Ero1α Cooperate for Efficient Disulfide Bond Formation in the Endoplasmic Reticulum

Human Ero1alpha is an endoplasmic reticulum (ER)-resident protein responsible for protein disulfide isomerase (PDI) oxidation. To clarify the molecular mechanisms underlying its function, we generated a panel of cysteine replacement mutants and analyzed their capability of: 1) complementing a temperature-sensitive yeast Ero1 mutant, 2) favoring oxidative folding in mammalian cells, 3) forming mixed disulfides with PDI and ERp44, and 4) adopting characteristic redox-dependent conformations. Our results reveal that two essential cysteine triads (Cys85-Cys94-Cys99 and Cys391-Cys394-Cys397) cooperate in electron transfer, with Cys94 likely forming mixed disulfides with PDI. Dominant negative phenotypes arise when critical residues within the triads are mutated (Cys394, Cys397, and to a lesser extent Cys99). Replacing the first cysteine in either triad (Cys85 or Cys391) generates mutants with weaker activity. In addition, mutating either Cys85 or Cys391, but not Cys397, reverts the dominant negative phenotype of the C394A mutant. These findings suggest that interactions between the two triads, dependent on Cys85 and Cys391, are important for Ero1alpha function, possibly stabilizing a platform for efficient PDI oxidation

A 5-Year-Old Case of Choroidal Neovascularization in Enhanced S-Cone Syndrome Treated with Ranibizumab

Introduction: We describe the youngest case of enhanced S-cone syndrome (ESCS) associated with choroidal neovascularization (CNV) successfully treated with intravitreal ranibizumab injections. Case Report: A 5-year-old boy presented with round-shaped fibrotic subretinal lesions in both eyes with surrounding subretinal fluid and progressive visual deterioration in the right eye. Fine foci of increased autofluorescence were observed along the arcades in both eyes. Fluorescein angiography revealed the presence of CNV in his right eye, and treatment with ranibizumab was initiated, with significant improvement in vision. Subsequent electroretinogram examination and genetic studies of the patient and his two younger siblings confirmed the diagnosis of ESCS. Conclusion: CNV has been reported to occur in different inherited retinal degenerations, including ESCS. Our experience confirms that treatment with ranibizumab in patients with CNV-complicated ESCS can be potentially vision-saving

Challenges to think about the city, territory, place. Shared experience between Mozambique, Brazil and Portugal

There are several challenges for those who today build, produce and reflect on cities, territories and places, given the different intersections, dimensions and possible scales, but also the specificities of each historical-geographical context. From a critical and reflective approach, based on the decolonial turn, and taking into account the growing urban socio-spatial and territorial inequalities, we seek to overcome imposed standards and recover local knowledge, highlighting the dynamics of bodies, subjects and groups, so often subjugated to thought and practice dominant in the Global North. Three researcher-teachers from Brazil, Mozambique and Portugal cross academic teaching, research and extension experiences in order to contribute to the construction of another way of apprehending and understanding the dimensions of spatial and territorial production, focusing on the province of Maputo, in particular the capital, Maputo, and the district of Boane. It starts from in loco research, project workshops and a close dialogue with local communities, but also from other teaching experiences, to identify and break, based on practice, pre-established paradigms and hierarchies. The notions of city, territory and place are discussed and revised, seeking to understand the current urgencies and absences, as well as other ways of overcoming the processes of segregation and inequality. Key-words: Maputo, urban theory, university tripod, decolonial critique.São vários os desafios de quem hoje constrói, produz e reflete sobre cidades, territórios e lugares, face às diferentes intersecções, dimensões e escalas possíveis, mas também às especificidades de cada contexto histórico-geográfico. A partir de uma abordagem crítica e reflexiva, baseada no giro decolonial, e atendendo às desigualdades urbanas socioespaciais e territoriais crescentes, procuramos a partir da prática, que aqui restituímos, ultrapassar padrões impostos e recuperar saberes locais, ressaltando a dinâmica dos corpos, sujeitos e grupos, tantas vezes subjugados ao pensamento e prática dominantes do Norte Global. Três pesquisadores-docentes do Brasil, Moçambique e Portugal cruzam assim experiências acadêmicas de ensino, pesquisa e extensão, por forma a contribuir para a construção de outro modo de apreender e compreender as dimensões da produção espacial e territorial, tendo como enfoque a província de Maputo, sobretudo a capital, Maputo, e o distrito de Boane. Partem-se de pesquisas in loco, de oficinas de projeto (extensionistas) e de um diálogo estreito com as comunidades locais, mas também de experiências de ensino outras, para identificar e romper paradigmas e hierarquias pré-estabelecidas. As noções de cidade, território e lugar são discutidas e revistas, procurando compreender as urgências e ausências atuais, bem como outras formas de superar os processos de segregação e desigualdade.   Palavras-chave: Maputo, teoria urbana, tripé universitário, crítica decolonial.  Peer Reviewe

Incidental Findings Suggestive of COVID-19 in Asymptomatic Patients Undergoing Nuclear Medicine Procedures in a High-Prevalence Region

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Influencia de la redación de la prescripción médica en la administración de medicamentos en horarios diferentes al prescripto

OBJETIVO: Analisar a influência da redação da prescrição médica na administração de medicamentos em horários diferentes do prescrito ocorridas em unidades de clínica médica de cinco hospitais brasileiros. MÉTODOS: Trata-se de estudo descritivo que utilizou dados secundários obtidos de uma pesquisa multicêntrica realizada em 2005. A amostra foi composta por 1084 doses de medicamentos administradas em horários diferentes do prescrito. RESULTADOS: Do total analisado, 96,2% apresentavam siglas e/ou abreviaturas; 7,8% apresentavam o registro do horário de administração incompleto e 4,8% destes registros estavam rasurados. Ainda, faltou o horário e/ou a freqüência de administração em 1,9% das prescrições. CONCLUSÃO: Com a implantação do sistema computadorizado de prescrições, associada à prática da educação permanente será possível minimizar a administração de medicamentos em horários diferentes do prescrito.OBJECTIVE: To evaluate the influence of the writing of medical orders on the administration of medications in medical units from five brazilian hospitals. METHODS: This descriptive study used a secondary analysis of data from a multicenter study conducted in 2005. the sample consisted of 1,084 medication orders that had been administered at the wrong schedule time. RESULTS: The great majority of medical orders (96.2%) had acronyms and/or abbreviations, 7.8% of them had incomplete schedules for administration of the medication, and 4.8% had been marked out. in addition, there was no schedule for the administration of the medication in 1.9% of the medical orders. CONCLUSION: Implementation of electronic prescribing and continuing education of health care providers can minimize the administration of medication at the wrong schedule time.OBJETIVO: Analizar la influencia de la redacción de la prescripción médica en la administración de medicamentos en horarios diferentes al prescripto ocurridas en unidades de clínica médica de cinco hospitales brasileños. MÉTODOS: Se trata de un estudio descriptivo que utilizó datos secundarios obtenidos de una investigación multicéntrica realizada en el 2005. La muestra estuvo compuesta por 1084 dosis de medicamentos administradas en horarios diferentes al prescripto. RESULTADOS: Del total analizado, el 96,2% presentaba siglas y/o abreviaturas; el 7,8% presentaba el registro del horario de administración incompleto y el 4,8% de estos registros estaban borrados. Aun más, faltó el horario y/o la frecuencia de administración en el 1,9% de las prescripciones. CONCLUSIÓN: Con la implantación del sistema computarizado de prescripciones, asociada a la práctica de la educación permanente será posible minimizar la administración de medicamentos en horarios diferentes al prescripto

Estratégia de Saúde da Família e prevalência de anemia em mulheres de uma região urbana de alto Índice de Desenvolvimento Humano

Introduction: Iron deficiency anemia, a public health problem. Objective: To analyze the hemoglobin value and the prevalence of anemia in women of reproductive age, mothers of children enrolled in municipal nurseries in a municipality and a high Human Development Index in the Southeast region, in addition to related factors, including being attended by the Family Health Strategy. Methods: A cross-sectional study, in a sample of 230 women aged 15 to 49 years. Collection of data through standardized form. Weight, height, and hemoglobin concentration were measured. Results: The prevalence of anemia in women was 9.6% and the mean hemoglobin concentration was 14.6g/dL. In the bivariate analysis, showed association with anemia the variables: age, having some disease, obstetric intercurrences in the child's study and as a protection factor and to be in the program Family Health Strategy. In the binary logistic analysis, only being attended by the Family Health Strategy demonstrated a significant protective effect against anemia (OR=0,391, p<0,05). Conclusion: Although there is a mild prevalence of anemia, according to World Health Organization criteria, in an urban area with a high Human Development Index, well-managed health care policies, the Family Health Strategy in particular, can contribute to further reduce the prevalence of this condition in the health of women of reproductive age.Introdução: Anemia ferropriva é um importante problema de saúde pública. Objetivo: Analisar o valor de hemoglobina e a prevalência de anemia em mulheres em idade reprodutiva, mães de filhos inscritos em creches municipais de um município com alto Índice de Desenvolvimento Humano, na região Sudeste, além de fatores a elas relacionados, incluindo serem atendidas pela Estratégia de Saúde da Família. Método: Trata-se de um estudo transversal com amostra constituída por 230 mulheres entre 15 e 49 anos de idade. A coleta de dados foi realizada por meio de formulário, abordando variáveis socioeconômicas e saúde materna. Verificaram-se: peso, estatura e dosagem da concentração da hemoglobina. Resultados: A prevalência de anemia nas mulheres foi de 9,6%, com média de concentração da hemoglobina de 14,6g/dL. Na análise bivariada, mostraram associação com anemia as variáveis: idade, ter alguma doença, intercorrências obstétricas na gestação da criança estudada e, como fator de proteção, estar no programa Estratégia de Saúde da Família. Na análise binária logística, apenas ser atendida pela Estratégia de Saúde da Família demonstrou ser significante efeito protetor frente à anemia (OR=0,391, p<0,05). Conclusão: Apesar de haver uma prevalência leve de anemia, pelo critério da Organização Mundial da Saúde, em área urbana de alto Índice de Desenvolvimento Humano, políticas de atenção à saúde e bem conduzidas, a Estratégia de Saúde da Família em particular, podem contribuir para reduzir ainda mais a prevalência desse agravo na saúde das mulheres em idade reprodutiva

Comparative Safety of Originator and Biosimilar Epoetin Alfa Drugs: An Observational Prospective Multicenter Study

Background: Erythropoiesis-stimulating agents (ESAs) are biological molecules approved for the treatment of anemia associated with chronic renal failure. Biosimilars were licensed for use in Europe in 2007. Aim: This study aimed to compare the safety profile of biosimilars with respect to the reference product in a nephrology setting. Methods: A prospective study was conducted in four Italian regions between 1 October 2013 and 30 June 2015. The study population included patients aged 65 18 years undergoing hemodialysis and treated with epoetins as per the clinical practice of the participating centers. The two comparison cohorts included patients treated with either an originator or a biosimilar epoetin alfa. Each patient was followed up until occurrence of any safety outcome of interest (grouped into three major categories), switch to a different ESA product, transplant or peritoneal dialysis, death, or end of the study period, whichever came first. Results: Overall, 867 subjects were included in the study (originator: N = 423; biosimilar: N = 444). Biosimilar users were older than originator users (median age of 76 vs 64 years, respectively), more frequently affected by arrhythmia (29.3 vs 22.5%), and less frequently candidates for transplantation (3.8 vs 18.2%). Cox-regression analysis showed no increase in risk of safety outcomes in biosimilar users, even after adjusting for confounding factors: 1.0 (95% confidence interval [CI] 0.7\u20131.3) for any outcomes; 1.1 (95% CI 0.7\u20131.8) for problems related to dialysis device; 0.9 (95% CI 0.6\u20131.5) for cardio- and cerebro-vascular conditions; 0.9 (95% CI 0.6\u20131.5) for infections. Conclusion: This study confirms the comparable safety profiles of originator and biosimilar epoetin alfa drugs when used in patients receiving dialysis

DECLINE OF PREVALENCE OF RESISTANCE ASSOCIATED SUBSTITUTIONS TO NS3 AND NS5A INHIBITORS AT DAA- FAILURE IN HEPATITIS C VIRUS IN ITALY OVER THE YEARS 2015 TO 2018

Background: A minority of patients fails to eliminate HCV and resistance-associated substitutions (RASs) are commonly detected at failure of interferon-free DAA regimens . Methods: Within the Italian network VIRONET-C, the prevalence of NS3/NS5A/NS5B RASs was retrospectively evaluated in patients who failed an EASL recommended DAA-regimen in 2015-2018 . The geno2pheno system and Sorbo MC et al. Drug Resistance Updates 2018 were used to infer HCV- genotype/subtype and predict drug resistance . The changes in prevalence of RASs over time were evaluated by chi-square test for trend, predictors of RASs at failure were analysed by logistic regression . Results: We included 386 HCV infected patients: 75% males, median age was 56 years (IQR 52-61), metavir fibrosis stage F4 in 76%; 106 (28%) were treatment- experienced: 91 (86%) with IFN-based treatments, 26 (25%) with DAAs. Patients with HIV and HBV coinfection were 10% (33/317) and 8% (6/72), respectively. HCV genotype was 1b in 122 pts (32%), 3 in 109 (28%), 1a in 97 (25%), 4 in 37 (10%), 2 in 21 (5%). DAA regimens were: LDV/SOF in 115 (30%), DCV/SOF in 103 (27%), 3D in 83 (21%), EBR/GRZ in 32 (8%), VEL/SOF in 29 (7%), GLE/PIB in 18 (5%) and 2D in 6 (2%); ribavirin was administered in 123 (32%) . The NS5A fasta-sequence was available for all patients, NS5B for 361 (94%), NS3 for 365 (95%) . According to the DAA failed the prevalence of any RASs was 90%, namely 80/135 (59%) in NS3, 313/359 (87%) in NS5A, 114/286 (40%) in NS5B . The prevalence of any RASs significantly declined from 2015 to 2018 (93% vs 70%, p=0.004): NS5A RASs from 90% to 72% (p=0 .29), NS3 RASs from 74% to 18% (p<0 .001), while NS5B RASs remained stable . Independent predictors of any RASs included advanced fibrosis (AOR 6.1, CI 95% 1.8-20.3, p=0 .004) and genotype (G2 vs G1a AOR 0 .03, CI 95% 0 .002- 0 .31, p=0 .004; G3 vs G1a AOR 0 .08, CI 95% 0 .01-0 .62, p=0 .02; G4 vs G1a AOR 0 .05, CI 95% 0 .006-0 .46, p=0 .008), after adjusting for age, previous HCV treatment and year of genotype . Notably, full activity was predicted for GLE/PIB in 75% of cases and for at least two components of VEL/SOF/VOX in 53% of cases, no case with full-resistance to either regimen was found . Conclusion: Despite decreasing prevalence over the years, RASs remain common at virological failure of DAA treatment, particularly in patients with the highest grade of liver fibrosis. The identification of RASs after failure could play a crucial role in optimizing retreatment strategies

Genetic determinants in a critical domain of ns5a correlate with hepatocellular carcinoma in cirrhotic patients infected with hcv genotype 1b

HCV is an important cause of hepatocellular carcinoma (HCC). HCV NS5A domain‐1 interacts with cellular proteins inducing pro‐oncogenic pathways. Thus, we explore genetic variations in NS5A domain‐1 and their association with HCC, by analyzing 188 NS5A sequences from HCV genotype‐1b infected DAA‐naïve cirrhotic patients: 34 with HCC and 154 without HCC. Specific NS5A mutations significantly correlate with HCC: S3T (8.8% vs. 1.3%, p = 0.01), T122M (8.8% vs. 0.0%, p < 0.001), M133I (20.6% vs. 3.9%, p < 0.001), and Q181E (11.8% vs. 0.6%, p < 0.001). By multivariable analysis, the presence of >1 of them independently correlates with HCC (OR (95%CI): 21.8 (5.7–82.3); p < 0.001). Focusing on HCC‐group, the presence of these mutations correlates with higher viremia (median (IQR): 5.7 (5.4–6.2) log IU/mL vs. 5.3 (4.4–5.6) log IU/mL, p = 0.02) and lower ALT (35 (30–71) vs. 83 (48–108) U/L, p = 0.004), suggesting a role in enhancing viral fitness without affecting necroinflammation. Notably, these mutations reside in NS5A regions known to interact with cellular proteins crucial for cell‐cycle regulation (p53, p85‐PIK3, and β‐ catenin), and introduce additional phosphorylation sites, a phenomenon known to ameliorate NS5A interaction with cellular proteins. Overall, these results provide a focus for further investigations on molecular bases of HCV‐mediated oncogenesis. The role of these NS5A domain‐1 mutations in triggering pro‐oncogenic stimuli that can persist also despite achievement of sustained virological response deserves further investigation

Fatality rate and predictors of mortality in an Italian cohort of hospitalized COVID-19 patients

Clinical features and natural history of coronavirus disease 2019 (COVID-19) differ widely among different countries and during different phases of the pandemia. Here, we aimed to evaluate the case fatality rate (CFR) and to identify predictors of mortality in a cohort of COVID-19 patients admitted to three hospitals of Northern Italy between March 1 and April 28, 2020. All these patients had a confirmed diagnosis of SARS-CoV-2 infection by molecular methods. During the study period 504/1697 patients died; thus, overall CFR was 29.7%. We looked for predictors of mortality in a subgroup of 486 patients (239 males, 59%; median age 71 years) for whom sufficient clinical data were available at data cut-off. Among the demographic and clinical variables considered, age, a diagnosis of cancer, obesity and current smoking independently predicted mortality. When laboratory data were added to the model in a further subgroup of patients, age, the diagnosis of cancer, and the baseline PaO2/FiO2 ratio were identified as independent predictors of mortality. In conclusion, the CFR of hospitalized patients in Northern Italy during the ascending phase of the COVID-19 pandemic approached 30%. The identification of mortality predictors might contribute to better stratification of individual patient risk