OBTENÇÃO DE SUPORTES HÍBRIDOS DE SÍLICA-MONOSSACARÍDEOS, COM POSSÍVEL UTILIZAÇÃO NA IMOBILIZAÇÃO DE PEROXIDASE.

A imobilização de enzimas em suportes híbridos mesoporosos (orgânico e inorgânico) atualmente apresenta um grande potencial, devido as melhores características do material quando comparadas a seus componentes separadamente. A síntese destes suportes híbridos pode ser realizada através da funcionalização pós-sintética (enxerto), a co-condensação ou na forma de organosílicas mesoporosas (ADAM et al., 2012).Diversos suportes inorgânicos de sílicas mesoporosas como, por exemplo, as sílicas SBA-15 e MCF já foram utilizados para a imobilização de peroxidase de raiz forte ou horseradishperoxidase (HRP), que é uma enzima de grande interesse em virtude das inúmeras possibilidades de aplicação (CAO et al., 2012). A HRP livre e/ou imobilizada apresenta diversas aplicações que unidas à sua elevada atividade, simplicidade na detecção e relativa estabilidade motiva o desenvolvimento de novos biocatalisadores que aumentem a estabilidade da mesma. Contudo a HRP possui um alto preço no mercado. A peroxidase de rabanete (Raphanus sativus) – RAP possui estrutura semelhante a HRP e pode ser utilizado como fonte alternativa de peroxidases. Considerando os estudos sobre imobilização de peroxidase extraída de rabanete (Raphanus sativus), verificou-se que não existem relatos na literatura quanto ao uso de suportes híbridos (inorgânico e orgânico) possibilitando um novo estudo que abarque a obtenção de novos suportes para a imobilização de enzimas econômicas com diversas aplicabilidades

SUPORTES HÍBRIDOS UTILIZADOS NA IMOBILIZAÇÃO DA PEROXIDASE DE RAIZ FORTE – HRP.

A imobilização de enzimas em suportes híbridos mesoporosos (orgânico e inorgânico) atualmente apresenta um grande potencial, devido as melhores características do material quando comparadas a seus componentes separadamente. A síntese destes suportes híbridos pode ser realizada através da funcionalização pós-sintética (enxerto), a co-condensação ou na forma de organosílicas mesoporosas (Adam et al., 2012).O método da co-condensação utiliza moléculas orgânicas como agentes moldes para a estrutura do material, onde ocorre a condensação do precursor na estrutura. Assim, esse método é bastante utilizado para a obtenção de suportes híbridos de sílica mesoporosos.O processo de imobilização pode ser realizado utilizando diferentes técnicas. Os métodos de adsorção física (ADS) e de ligação covalente (LC) são os mais utilizados. Segundo Singh e colaboradores os métodos por adsorção física e ligação covalente podem reduzir ou evitar a lixiviação da enzima. A enzima pode-se ligar ao suporte orgânico ou inorgânico através de interações adicionais covalentes ou não covalentes, que ocasiona na diminuição da flexibilidade estrutural da enzima e uma maior rigidez a enzima imobilizada, reduzindo a possibilidade de desnaturação (Singh et al., 2013).A peroxidase de raiz forte ou horseradish peroxidase (HRP) apresenta diversificadas aplicações na indústria, desde a descoloração de corantes sintéticos até o desenvolvimento de biossensores (Mohamed et al., 2013), na área clínica e ambiental . Estas possibilidades de aplicação unidas à sua elevada atividade, simplicidade na detecção de produtos e relativa estabilidade leva a desenvolver novos métodos ou suportes que aumentem a estabilidade desta enzima. Suportes inorgânicos de sílicas mesoporosas como, por exemplo, as sílicas SBA-15 e MCF já foram utilizados para a imobilização da HRP com bons resultados (Cao et al., 2013). O objetivo deste trabalho foi desenvolver suportes híbridos que apresentem em sua composição sílica e alginato de sódio com posterior aplicação na imobilização de HRP por ADS e LC

Aplicações analíticas de antocianinas extraídas de frutos nativos do semi-árido baiano.

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Design and baseline characteristics of the finerenone in reducing cardiovascular mortality and morbidity in diabetic kidney disease trial

Background: Among people with diabetes, those with kidney disease have exceptionally high rates of cardiovascular (CV) morbidity and mortality and progression of their underlying kidney disease. Finerenone is a novel, nonsteroidal, selective mineralocorticoid receptor antagonist that has shown to reduce albuminuria in type 2 diabetes (T2D) patients with chronic kidney disease (CKD) while revealing only a low risk of hyperkalemia. However, the effect of finerenone on CV and renal outcomes has not yet been investigated in long-term trials. Patients and Methods: The Finerenone in Reducing CV Mortality and Morbidity in Diabetic Kidney Disease (FIGARO-DKD) trial aims to assess the efficacy and safety of finerenone compared to placebo at reducing clinically important CV and renal outcomes in T2D patients with CKD. FIGARO-DKD is a randomized, double-blind, placebo-controlled, parallel-group, event-driven trial running in 47 countries with an expected duration of approximately 6 years. FIGARO-DKD randomized 7,437 patients with an estimated glomerular filtration rate >= 25 mL/min/1.73 m(2) and albuminuria (urinary albumin-to-creatinine ratio >= 30 to <= 5,000 mg/g). The study has at least 90% power to detect a 20% reduction in the risk of the primary outcome (overall two-sided significance level alpha = 0.05), the composite of time to first occurrence of CV death, nonfatal myocardial infarction, nonfatal stroke, or hospitalization for heart failure. Conclusions: FIGARO-DKD will determine whether an optimally treated cohort of T2D patients with CKD at high risk of CV and renal events will experience cardiorenal benefits with the addition of finerenone to their treatment regimen. Trial Registration: EudraCT number: 2015-000950-39; ClinicalTrials.gov identifier: NCT02545049

Mortality from gastrointestinal congenital anomalies at 264 hospitals in 74 low-income, middle-income, and high-income countries: a multicentre, international, prospective cohort study

Summary Background Congenital anomalies are the fifth leading cause of mortality in children younger than 5 years globally. Many gastrointestinal congenital anomalies are fatal without timely access to neonatal surgical care, but few studies have been done on these conditions in low-income and middle-income countries (LMICs). We compared outcomes of the seven most common gastrointestinal congenital anomalies in low-income, middle-income, and high-income countries globally, and identified factors associated with mortality. Methods We did a multicentre, international prospective cohort study of patients younger than 16 years, presenting to hospital for the first time with oesophageal atresia, congenital diaphragmatic hernia, intestinal atresia, gastroschisis, exomphalos, anorectal malformation, and Hirschsprung’s disease. Recruitment was of consecutive patients for a minimum of 1 month between October, 2018, and April, 2019. We collected data on patient demographics, clinical status, interventions, and outcomes using the REDCap platform. Patients were followed up for 30 days after primary intervention, or 30 days after admission if they did not receive an intervention. The primary outcome was all-cause, in-hospital mortality for all conditions combined and each condition individually, stratified by country income status. We did a complete case analysis. Findings We included 3849 patients with 3975 study conditions (560 with oesophageal atresia, 448 with congenital diaphragmatic hernia, 681 with intestinal atresia, 453 with gastroschisis, 325 with exomphalos, 991 with anorectal malformation, and 517 with Hirschsprung’s disease) from 264 hospitals (89 in high-income countries, 166 in middleincome countries, and nine in low-income countries) in 74 countries. Of the 3849 patients, 2231 (58·0%) were male. Median gestational age at birth was 38 weeks (IQR 36–39) and median bodyweight at presentation was 2·8 kg (2·3–3·3). Mortality among all patients was 37 (39·8%) of 93 in low-income countries, 583 (20·4%) of 2860 in middle-income countries, and 50 (5·6%) of 896 in high-income countries (p<0·0001 between all country income groups). Gastroschisis had the greatest difference in mortality between country income strata (nine [90·0%] of ten in lowincome countries, 97 [31·9%] of 304 in middle-income countries, and two [1·4%] of 139 in high-income countries; p≤0·0001 between all country income groups). Factors significantly associated with higher mortality for all patients combined included country income status (low-income vs high-income countries, risk ratio 2·78 [95% CI 1·88–4·11], p<0·0001; middle-income vs high-income countries, 2·11 [1·59–2·79], p<0·0001), sepsis at presentation (1·20 [1·04–1·40], p=0·016), higher American Society of Anesthesiologists (ASA) score at primary intervention (ASA 4–5 vs ASA 1–2, 1·82 [1·40–2·35], p<0·0001; ASA 3 vs ASA 1–2, 1·58, [1·30–1·92], p<0·0001]), surgical safety checklist not used (1·39 [1·02–1·90], p=0·035), and ventilation or parenteral nutrition unavailable when needed (ventilation 1·96, [1·41–2·71], p=0·0001; parenteral nutrition 1·35, [1·05–1·74], p=0·018). Administration of parenteral nutrition (0·61, [0·47–0·79], p=0·0002) and use of a peripherally inserted central catheter (0·65 [0·50–0·86], p=0·0024) or percutaneous central line (0·69 [0·48–1·00], p=0·049) were associated with lower mortality. Interpretation Unacceptable differences in mortality exist for gastrointestinal congenital anomalies between lowincome, middle-income, and high-income countries. Improving access to quality neonatal surgical care in LMICs will be vital to achieve Sustainable Development Goal 3.2 of ending preventable deaths in neonates and children younger than 5 years by 2030

Non-contact specular microscopy in aphakic and pseudophakic dogs

Cataract is one of the most common ocular diseases in dogs, and phacoemulsification is considered its treatment of choice. Posterior capsular opacity (PCO) is a frequent complication and may occur weeks or months after the surgery. It is known that intraocular lenses (IOL) used for the correction of post-operative hypermetropia are an adjunctive aid in the prevention of PCOs, mainly the foldable acrylics designs with square and truncated edges. However, IOL placement may result in endothelial cell loss. This study evaluated the morphology and cellularity of the endothelium by non-contact specular microscopy in aphakic and pseudophakic dogs. Endothelial cell density, endothelial cell area, coefficient of variation and hexagonal cells percent were analyzed for each group and between groups. During all evaluation periods, endothelial cell density did not alter in any group. From days 7 to 30, endothelial cell density was higher in pseudophakic dogs. However, values returned to normal and, by the end of the study, no significant difference was observed. Cell area and variation coefficient did not differ in each group or between groups. Hexagonal cells percent significantly reduced after phacoemulsification in both groups. With time, this condition returned to initial values. When both groups were compared, hexagonal cells percent were decreased in the pseudophakic group, immediately after the surgeries.A catarata está entre as afecções oculares mais frequentes em cães, sendo a facoemulsificação o método que melhor se adequa à sua terapia. A opacidade de cápsula posterior (OCP) encontra-se entre as complicações mais comuns e se manifesta em semanas ou meses após a facectomia. Lentes intraoculares, empregadas na correção da hipermetropia pós-cirúrgica, contribuem para a prevenção das OCPs, especialmente as acrílicas dobráveis, com bordas truncadas ou quadradas. Entretanto, a sua implantação pode resultar na perda de células endoteliais. Com a pesquisa, propuseram-se avaliar a morfologia e a celularidade endoteliais, a microscopia especular de não contato, em pacientes afácicos e em pseudofácicos. Foram analisados a densidade celular, a área celular, o coeficiente de variação e o percentual de células hexagonais inter e entre os grupos. Ao longo dos momentos da avaliação, a densidade das células endoteliais não se alterou significativamente dentro dos grupos. Na comparação entre os grupos, ela foi maior nos pseudofácicos, em períodos que variaram de 7 a 30 dias, entretanto retornaram a valores próximos aos iniciais, tornando a diferença entre os dois grupos não significativa ao final da avaliação. Relativamente à área celular e ao coeficiente de variação, não houve diferença significativa, inter e entre os grupos. Houve diferença dentro dos grupos relativamente à hexagonalidade, com diminuição percentual após a facoemulsificação. As condições retornaram ao basal. Na comparação entre pacientes pseudofácicos e afácicos, houve diminuição nos pseudofácicos imediatamente à cirurgia.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Altered behavioural response to acute stress in mice lacking cellular prion protein

Although many studies have investigated the function of cellular prion protein (PrPc), its physiologic role remains elusive. PrPc null mice have been reported to develop normally and to show normal performance in most behavioural tests. In the present study we investigated whether this also holds true after episodes of acute stress. PrPc gene ablated (Prnp0/0) and wild-type mice were subjected to restraint stress, electric foot shock, or swimming and compared with non-stressed animals. Immediately after the stressful situation, the anxiety levels and locomotion of the animals were measured using plus-maze and open-field tests. Among non-stressed animals, there was no significant difference in performance between Prnp0/0 and wild type animals in either test. However, after acute stress provoked by a foanot shock or a swimming trial, Prnp0/0 animals showed a significant decrease in anxiety levels when compared with control animals. Moreover, after the swimming test, knockout mice presented decreased locomotion when compared to wild-type mice. Because of this observation, we also assessed both types of mice in a forced swimming test with the objective of better evaluating muscle function and found that Prnp0/0 animals presented reduced forced swimming capacity when compared to controls. As far as we know, this is the first report suggesting that cellular prion protein is involved in modulation of anxiety or muscular activity after acute psychic or physical stress