Glyoxyl-Activated Agarose as Support for Covalently Link Novo-Pro D: Biocatalysts Performance in the Hydrolysis of Casein

© 2020 by the authors.This study aimed to evaluate the performance of a commercial protease (Novo-Pro D (NPD)), both in soluble and immobilized forms, in the hydrolysis of proteins (using casein as model protein). Immobilization of the protease NPD on 6% agarose activated with glyoxyl groups for 24 h at 20 °C and pH 10.0 allowed preparing immobilized biocatalyst with around 90% immobilization yield, 92% recovered activity versus small substrate, and a thermal stability 5.3-fold higher than the dialyzed soluble enzyme at 50 °C and pH 8.0. Immobilization times longer than 24 h lead to a decrease in the recovered activity and did not improve the biocatalyst stability. At 50 °C and pH 6.5, the immobilized NPD was around 20-fold more stable than the dialyzed soluble protease. Versus casein, the immobilized NDP presented a 10% level of activity, but it allowed hydrolyzing casein (26 g/L) at 50 °C and pH 6.5 up to a 40% degree of hydrolysis (DH) after 2 h reaction, while under the same conditions, only a 34% DH was achieved with soluble NPD. In addition, the immobilized NPD showed good reusability, maintaining the DH of casein for at least ten 2h-reaction batches.This research was funded by National Council for Scientific and Technology Development (CNPq), grant numbers 141487/2016-0, 422131/2016-4; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brazil (CAPES), Finance Code 001 and Spanish Ministerio de Ciencia e Innovación, grant number CTQ2017-86170-R.Peer reviewe

Volumetric Blood Flow in Transjugular Intrahepatic Portosystemic Shunt Revision Using 3‐Dimensional Doppler Sonography

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/135560/1/jum2015342257.pd

The Role of Oxidative Stress and Lipid Peroxidation in Ventricular Remodeling Induced by Tobacco Smoke Exposure after Myocardial Infarction

OBJECTIVE: To evaluate the roles of oxidative stress and lipid peroxidation in the ventricular remodeling that is induced by tobacco smoke exposure after myocardial infarction.METHODS: After induced myocardial infarction, rats were allocated into two groups: C (control, n=25) and ETS (exposed to tobacco smoke, n=24). After 6 months, survivors were submitted to echocardiogram and biochemical analyses.RESULTS: Rats in the ETS group showed higher diastolic (C = 1.52 +/- 0.4 mm(2), ETS = 1.95 +/- 0.4 mm(2); p=0.032) and systolic (C = 1.03 +/- 0.3, ETS = 1.36 +/- 0.4 mm(2)/g; p=0.049) ventricular areas, adjusted for body weight. The fractional area change was smaller in the ETS group (C = 30.3 +/- 10.1 %, ETS = 19.2 +/- 11.1 %; p=0.024) and E/A ratios were higher in ETS animals (C = 2.3 +/- 2.2, ETS = 5.1 +/- 2.5; p=0.037). ETS was also associated with a higher water percentage in the lung (C = 4.8 (4.3-4.8), ETS = 5.5 (5.3-5.6); p=0.013) as well as higher cardiac levels of reduced glutathione (C = 20.7 +/- 7.6 nmol/mg of protein, ETS = 40.7 +/- 12.7 nmol/mg of protein; p=0.037) and oxidized glutathione (C = 0.3 +/- 0.1 nmol/g of protein, ETS = 0.9 +/- 0.3 nmol/g of protein; p=0.008). No differences were observed in lipid hydroperoxide levels (C = 0.4 +/- 0.2 nmol/mg of tissue, ETS = 0.1 +/- 0.1 nmol/mg of tissue; p=0.08).CONCLUSION: In animals exposed to tobacco smoke, oxidative stress is associated with the intensification of ventricular re-remodeling after myocardial infarction

Effect of pruning strategy on 'Syrah' bud necrosis and fruitfulness in Brazilian subtropical Southeast

The change of wine grape harvest from wet season (summer) to dry season (winter) by changing the pruning management has improved quality of wines produced in the Brazilian Southeast. However, the vines need to be spur pruned twice a year, i.e. with a 1st pruning in August (winter pruning) for a vegetative cycle during the hot and wet summer, and a 2nd pruning in January (summer pruning) for a productive cycle during the cold and dry season. This double pruning strategy is made necessary by the fact that latent buds developed during the dry season cycle are not fruitful to support a productive cycle in the following year. This histological study, performed in the South of Minas Gerais State (Brazil), showed that annual single pruning done in the wet season (in January) displayed a high rate of necrosis on primary and secondary buds (bud necrosis – BN). In April, 99 days after summer pruning (DASP), the rates of BN were 40 % and 50 % at basal and apical node positions, respectively, reaching 80 % of BN in December (322 DASP). As a consequence of BN, bud potential fertility was drastically reduced from 0.5 inflorescence primordial (IP) per bud (in July) to 0.06 (in December) and bud burst in the next cycle from secondary and tertiary bud axes. Vines managed by double pruning system (submitted to summer and winter pruning) displayed a much higher fruitfulness potential, i.e. 1.46 IP per bud in December (112 days after winter pruning) and limited BN occurrence (20 %). On single pruned vines, we also observed a significant decrease of starch content in canes, trunks and roots. Internal bud anatomy showed that a random cell breakdown started 70 days DASP. At 211 DASP, all buds showed a large starch granule concentration, raphides and crystals of calcium oxalate inside idioblasts of leaf primordia and also in cortical parenchyma of the vegetative axis. The bud starch content was increased and a positive correlation between necrosis and starch accumulation was observed. The impact of carbohydrate availability on bud necrosis development was discussed. This study showed that the necrosis development towards secondary and tertiary axis of the dry season buds is the main reason of unfruitfulness in the vineyards managed by single pruning in the wet season, making the double pruning compulsory

The impact of hypoxia-inducible factors in the pathogenesis of kidney diseases: a link through cell metabolism

Kidneys are sensitive to disturbances in oxygen homeostasis. Hypoxia and activation of the hypoxia-inducible factor (HIF) pathway alter the expression of genes involved in the metabolism of renal and immune cells, interfering with their functioning. Whether the transcriptional activity of HIF protects the kidneys or participates in the pathogenesis of renal diseases is unclear. Several studies have indicated that HIF signaling promotes fibrosis in experimental models of kidney disease. Other reports showed a protective effect of HIF activation on kidney inflammation and injury. In addition to the direct effect of HIF on the kidneys, experimental evidence indicates that HIF-mediated metabolic shift activates inflammatory cells, supporting the HIF cascade as a link between lung or gut damage and worsening of renal disease. Although hypoxia and HIF activation are present in several scenarios of renal diseases, further investigations are needed to clarify whether interfering with the HIF pathway is beneficial in different pathological contexts

Reasons and factors related to non-adherence to breast and cervical cancer screening in primary health care in São José do Rio Preto - SP after the COVID-19 pandemic

Objective: To identify the reasons and factors associated with non-adhence of mammogram and Pap smear. Methods: This cross-sectional observational study was conducted through interviews with women between 25 and 69 years-old in basic health units. Results: 441 women were included in the study, 404 of whom were eligible for cervical cancer screening and 208 for breast cancer screening. Mammogram adherence was associated with educational level (p<0.001), family history of breast cancer (p=0.047), self-assessment of health status (p<0.001) and prior knowledge about age group (p<0.001) and frequency (p<0.001) of screening recommended by the Ministry of Health. However, the decision of undergo cervical cancer screening exam was associated with age (p=0.012), self-assessment of health status (p<0.001) and prior knowledge about age group (p<0.001) and frequency (p=0.027) of exam. In both groups, the main reasons for non-adherence to screening were: lack of time, difficulty scheduling appointments and/or tests, and the SARS-Cov-2 pandemic. Conclusion: Socioeconomic factors and health education influence the decision to participate in the secondary prevention of these neoplasms. Thus, health policies should be intensified addressing these topics to increase the adherence rate to exams.Objetivo: Identificar os motivos e os fatores associados à não realização da mamografia e do exame de Papanicolaou. Métodos: Este estudo observacional transversal foi conduzido por meio de entrevistas com mulheres de 25 a 69 anos em unidades básicas de saúde. Resultados: Foram incluídas 441 mulheres no estudo, sendo 404 elegíveis para o rastreamento do câncer de colo do útero e 208 para o rastreamento de câncer de mama. A realização da mamografia foi associada ao nível de educação (p<0,001), histórico familiar de câncer de mama (p=0,047), autoavaliação do estado de saúde (p<0,001) e conhecimento prévio sobre a faixa etária (p<0,001) e frequência (p<0,001) do rastreamento preconizada pelo Ministério da Saúde. Contudo, a adesão ao exame de rastreamento do câncer cervical foi associada à idade (p=0,012), autoavaliação do estado de saúde (p<0,001) e conhecimento prévio sobre a faixa etária (p<0,001) e frequência (p=0,027) do exame. Em ambos os grupos, os principais motivos da não adesão ao rastreamento foram: falta de tempo, dificuldade no agendamento na consulta e/ou exame e a pandemia de SARS-Cov-2. Conclusão: Fatores socioeconômicos e educação em saúde influenciam a decisão de participar da prevenção secundária dessas neoplasias. Desse modo, as políticas de saúde devem ser intensificadas, abordando esses tópicos para elevar as taxas de adesão aos exames

The genetic diagnosis of rare endocrine disorders of sex development and maturation : a survey among Endo-ERN centres

Differences of sex development and maturation (SDM) represent a heterogeneous puzzle of rare conditions with a large genetic component whose management and treatment could be improved by an accurate classification of underlying molecular conditions, and next-generation sequencing (NGS) should represent the most appropriate approach. Therefore, we conducted a survey dedicated to the use and potential outcomes of NGS for SDM disorders diagnosis among the 53 health care providers (HCP) of the European Reference Network for rare endocrine conditions. The response rate was 49% with a total of 26 HCPs from 13 countries. All HCPs, except 1, performed NGS investigations for SDM disorders on 6720 patients, 3764 (56%) with differences of sex development (DSD), including 811 unexplained primary ovarian insufficiency, and 2956 (44%) with congenital hypogonadotropic hypogonadism (CHH). The approaches varied from targeted analysis of custom gene panels (range: 11-490 genes) in 81.5% of cases or whole exome sequencing with the extraction of a virtual panel in the remaining cases. These analyses were performed for diagnostic purposes in 21 HCPs, supported by the National Health Systems in 16 cases. The likelihood of finding a variant ranged between 7 and 60%, mainly depending upon the number of analysed genes or criteria used for reporting, most HCPs also reporting variants of uncertain significance. These data illustrate the status of genetic diagnosis of DSD and CHH across Europe. In most countries, these analyses are performed for diagnostic purposes, yielding highly variable results, thus suggesting the need for harmonization and general improvements of NGS approaches.publishersversionPeer reviewe

Advances in Carcinogenic Metal Toxicity and Potential Molecular Markers

Metal compounds such as arsenic, cadmium, chromium, cobalt, lead, mercury, and nickel are classified as carcinogens affecting human health through occupational and environmental exposure. However, the underlying mechanisms involved in tumor formation are not well clarified. Interference of metal homeostasis may result in oxidative stress which represents an imbalance between production of free radicals and the system’s ability to readily detoxify reactive intermediates. This event consequently causes DNA damage, lipid peroxidation, protein modification, and possibly symptomatic effects for various diseases including cancer. This review discusses predominant modes of action and numerous molecular markers. Attention is paid to metal-induced generation of free radicals, the phenomenon of oxidative stress, damage to DNA, lipid, and proteins, responsive signal transduction pathways with major roles in cell growth and development, and roles of antioxidant enzymatic and DNA repair systems. Interaction of non-enzymatic antioxidants (carotenoids, flavonoids, glutathione, selenium, vitamin C, vitamin E, and others) with cellular oxidative stress markers (catalase, glutathione peroxidase, and superoxide dismutase) as well as certain regulatory factors, including AP-1, NF-κB, Ref-1, and p53 is also reviewed. Dysregulation of protective pathways, including cellular antioxidant network against free radicals as well as DNA repair deficiency is related to oncogenic stimulation. These observations provide evidence that emerging oxidative stress-responsive regulatory factors and DNA repair proteins are putative predictive factors for tumor initiation and progression