Analysis of Germin-like protein genes family in Vitis vinifera (VvGLPs) using various in silico approaches / Análise da família de genes de proteínas semelhantes a germin em Vitis vinifera (VvGLPs) usando várias abordagens in silico

Germin-like proteins (GLPs) play an important role against various stresses. Vitis vinifera L. genome contains 7 GLPs; many of them are functionally unexplored. However, the computational analysis may provide important new insight into their function. Currently, physicochemical properties, subcellular localization, domain architectures, 3D structures, N-glycosylation & phosphorylation sites, and phylogeney of the VvGLPs were investigated using the latest computational tools. Their functions were predicted using the Search tool for the retrieval of interacting genes/proteins (STRING) and Blast2Go servers. Most of the VvGLPs were extracellular (43%) in nature but also showed periplasmic (29%), plasma membrane (14%), and mitochondrial- or chloroplast-specific (14%) expression. The functional analysis predicted unique enzymatic activities for these proteins including terpene synthase, isoprenoid synthase, lipoxygenase, phosphate permease, receptor kinase, and hydrolases generally mediated by Mn+ cation. VvGLPs showed similarity in the overall structure, shape, and position of the cupin domain. Functionally, VvGLPs control and regulate the production of secondary metabolites to cope with various stresses. Phylogenetically VvGLP1, -3, -4, -5, and VvGLP7 showed greater similarity due to duplication while VvGLP2 and VvGLP6 revealed a distant relationship. Promoter analysis revealed the presence of diverse cis-regulatory elements among which CAAT box, MYB, MYC, unnamed-4 were common to all of them. The analysis will help to utilize VvGLPs and their promoters in future food programs by developing resistant cultivars against various biotic (Erysiphe necator and in Powdery Mildew etc.) and abiotic (Salt, drought, heat, dehydration, etc.) stresses.

As proteínas do tipo germin (GLPs) desempenham um papel importante contra vários estresses. O genoma de Vitis vinifera L. contém 7 GLPs; muitos deles são funcionalmente inexplorados. No entanto, a análise computacional pode fornecer informações importantes sobre sua função. Atualmente, as propriedades físico-químicas, localização subcelular, arquitetura de domínio, estruturas 3D, sítios de N-glicosilação e fosforilação e estudos filogenéticos dos VvGLPs foram conduzidos usando as ferramentas computacionais mais recentes. Suas funções foram previstas usando a ferramenta Search para recuperação de genes/proteínas em interação (STRING) e servidores Blast2Go. A maioria dos VvGLPs são extracelulares (43%) na natureza, mas também mostraram expressão periplasmática (29%), na membrana plasmática (14%) e específica para mitocôndrias ou cloroplastos (14%). A análise funcional previu atividades enzimáticas únicas para essas proteínas, incluindo terpeno sintase, isoprenoide sintase, lipoxigenase, fosfato permease, receptor quinase e hidrolases geralmente mediadas por cátion Mn +. VvGLPs mostraram similaridade na estrutura geral, forma e posição do domínio cupin. Funcionalmente, os VvGLPs controlam e regulam a produção de metabólitos secundários para lidar com vários estresses. Filogeneticamente, VvGLP1, -3, -4, -5 e VvGLP7 mostraram maior similaridade devido à duplicação, enquanto VvGLP2 e VvGLP6 revelaram uma relação distante. A análise do promotor revelou a presença de diversos elementos cis-reguladores, entre os quais CAAT box, MYB, MYC, sem nome-4, sendo comum a todos eles. A análise ajudará a utilizar VvGLPs e seus promotores em programas alimentares futuros, desenvolvendo cultivares resistentes contra vários estresses bióticos (Erysiphe necator e no oídio, etc.) e abióticos (sal, seca, calor, estresse hídrico, etc.).

Análises bromatológicas de missô artesanal / Bromatological analysis of handmade misso

Objetivo ­ Realizar análises bromatológicas de umidade, proteínas, pH e cinzas em missôs artesanais. A fermentação é um processo biotecnológico que tem sido utilizado para modificar e produzir alimentos desde a antiguidade. Nas últimas duas décadas, o interesse nos efeitos benéficos dos fermentados na saúde humana aumentou e tornou essa categoria de alimentos cada vez mais popular principalmente no Oriente. No mercado há uma ampla variedade de pastas à base de soja fermentada por microorganismos sendo conhecido popularmente como missô. Métodos ­ As análises realizadas foram secagem direta em estufa a 105°C graus para determinação da umidade (%) e calcinação em mufla para cinzas (%), determinação de pH por meio do peagâmetro e análise de proteínas através do teste de Biureto. Resultados ­ No presente estudo as amostras obtiveram um teor de umidade entre 52,71% a 60,48%, teor de cinzas variando de 1,12% a 22,7%, pH entre 5,35 e 8,68, e um teor de proteínas variando de 11,1% a 13,2%. Discussão ­ Foi interpretado e comparado os resultados obtidos com as análises de outros estudos, além disso, apontado algumas questões do campo bromatológico das pesquisas dos estudos comparados e as limitações do presente trabalho. Conclusão ­ O processo fermentativo de alimentos com microorganismos resulta em um produto diferenciado que pode ser benéfico a saúde com diferentes características organolépticas. Nossos resultados foram parcialmente semelhantes com outras pesquisas sendo que

Propiedades nutritivas y tecno funcionales de barras de pseudocereales adicionadas con soya, mango y granada / Nutritional and techno functional properties of pseudocereal bars added with soy, mango and pomegranate

Las barras de cereal (BC), se comercializan como un snack saludable, no obstante, su calidad nutricional es baja. Una alternativa para mejorar esto, es la incorporación de ingredientes como pseudocereales, germinados y subproductos de fruta. Objetivo. Evaluar el contenido nutricional y propiedades tecnofuncionales de una barra de cereal formulada a partir de pseudocereales, germinados de soya y subproductos del procesamiento de frutas. Materiales y Métodos. Se desarrollaron 6 formulaciones (F0-F5). Se determinó el contenido de proteína y fibra cruda, se seleccionó la formulación que presentó el mayor contenido de estos componentes. A la BC seleccionada se le determinó la digestibilidad in-vitro de la proteína, las propiedades tecnofuncionales potencial prebiótico y actividad inhibitoria de ECA-I. Resultados. La formulación seleccionada fue F1 (13,6 g/100 g p.s. proteína y 13,1 g/100 g p.s. fibra cruda). La digestibilidad de la proteína fue del 69 %, el cual es cercano a valores reportados para algunos componentes de la BC. La capacidad de hinchamiento y retención de agua fue 2,55 ml/g; 12,74 %, respectivamente. El crecimiento de L. brevis en medio MRS modificado con BC no presentó diferencias estadísticas con el medio control, indicando el potencial prebiótico presente en la BC. La barra de cereal tuvo un 39% de actividad inhibitoria de ECA-I, demostrando la acción de los compuestos bioactivos posiblemente liberados durante la digestión de la BC. Conclusión. La formulación desarrollada presenta propiedades funcionales importantes y podría generar beneficios para la salud(AU)

Introduction. Cereal bars (CB) are marketed as a healthy snack; however, their nutritional quality is low. An alternative to improve this is the incorporation of ingredients such as soybean sprouts, which have a higher protein content than some seeds; as well as fruit by-products that contain important bioactive compounds. Objective. To evaluate the nutritional content and techno-functional properties of a cereal bar formulated from pseudocereals, soybean sprouts, and fruit processing by-products. Materials and Methods. 6 formulations (F0-F5) were developed. The content of protein and crude fiber was determined, the formulation that presented the highest content of these components was selected. The in-vitro digestibility of the protein, the technofunctional properties, potential prebiotic and inhibitory activity of ACE-I were determined for the selected BC. Results. The selected formulation was F1 (13.6g/100g p.s. protein and 13.1g/100 g p.s. crude fiber). Protein digestibility was 69%, which is close to reported values for some CB components. The swelling and water retention capacity was 2.55 ml/g; 12.74%, respectively. The growth of L. brevis in modified MRS medium with CB did not present statistical differences with the control medium, indicating the prebiotic potential present in CB. The cereal bar had 39% ACE-I inhibitory activity, demonstrating the action of bioactive compounds possibly released during CB digestion. Conclusion. The developed formulation has important functional properties and could generate health benefits(AU)

Proteínas oxidadas de origen animal y su impacto sobre la salud intestinal / Oxidized proteins of animal origin and their impact on intestinal health / Proteínas oxidadas de origem animal e seu impacto na saúde intestinal

Los alimentos de origen animal como la carne de pollo, res, pescado y cerdo poseen una amplia demanda en todo el mundo debido, entre otros aspectos, a su valor nutricional, asociado al alto contenido proteico. No obstante, este tipo de proteínas son susceptibles de sufrir reacciones de oxidación, las cuales pueden mediar procesos de fragmentación, agregación, pérdida de solubilidad, funcionalidad y digestibilidad proteica; eventos implicados en la pérdida de su valor nutricional. En este sentido, las proteínas agrega­das tienden a no ser digeridas en el tracto gastrointestinal y acumularse en el intestino (colon), donde la microbiota colónica las degrada a productos mutagénicos como fenol y p-cresol, lo que incrementa el riesgo de cáncer colorrectal. Por otra parte, los ami­noácidos o péptidos oxidados liberados en la digestión podrían incorporarse en las vías de señalización celular intestinal y favorecer o exacerbar procesos intestinales crónicos como colon irritable o enfermedad de Crohn. Debido al gran interés de esta temática en los últimos años, el objetivo de esta revisión es realizar una descripción general del impacto de proteínas oxidadas de origen animal sobre la salud intestinal.

Animal foods such as chicken, beef, fish and pork are in wide demand throughout the world due, among other things, to their nutritional value, associated with their high protein content. However, this type of protein is susceptible to oxidation reactions, which can mediate processes of fragmentation, aggregation, loss of solubility, functionality, and protein digestibility, which are events involved in the loss of their nutritional value. In this sense, aggregated proteins tend not to be digested in the gastrointestinal tract and accumulate in the intestine (colon), where the colonic microbiota degrades them into mutagenic products such as phenol and p-cresol, which increases the risk of colorectal cancer. On the other hand, the oxidized amino acids or peptides released in digestion could be incorporated into intestinal cell signaling pathways and favor or exacerbate chronic intestinal processes such as irritable bowel syndrome or Crohn's disease. Due to the great interest in this topic in recent years, the objective of this review is to provide a general overview of the impact of oxidized proteins of animal origin on intestinal health.

Alimentos de origem animal como frango, carne bovina, peixe e carne suína são muito procurados em todo o mundo devido, entre outros fatores, ao seu valor nutricional, associado ao seu alto teor de proteínas. No entanto, esse tipo de proteína é suscetível a reações de oxidação, que podem mediar processos de fragmentação, agregação, perda de solubilidade, funcionalidade e digestibilidade da proteína; eventos envolvidos na perda de seu valor nutritivo. Nesse sentido, as proteínas agregadas tendem a não ser digeridas no trato gastrointestinal e se acumulam no intestino (cólon), onde a microbiota colônica as degrada em produtos mutagênicos como fenol e p - cresol, aumentando o risco de câncer colorretal. Por outro lado, os aminoácidos ou peptídeos oxidados liberados na digestão poderiam ser incorporados às vias de sinalização das células intestinais e favorecer ou exacerbar processos intestinais crônicos, como a síndrome do intestino irritável ou a doença de Crohn. Devido ao grande interesse neste tema nos últimos anos, o objetivo desta revisão é fornecer uma descrição geral do impacto das proteínas oxidadas de origem animal na saúde intestinal.

Fisiología respiratoria: fisiología del surfactante pulmonar / Physiology of pulmonary surfactant

El proceso de respiración y el intercambio gaseoso requiere la interacción de variadas fuerzas en los distintos tejidos y órganos involucrados. La tensión superficial a nivel alveolar provocaría colapso de dichas estructuras de no ser por las características del surfactante que lo recubre. Revisaremos en este articulo la fisiología involucrada en su estructura física, producción y efectos pulmonares.

The process of breathing and gas exchange requires the interaction of various forces in the different tissues and organs involved. The surface tension at the alveolus would cause collapse of these structures without of the surfactant that covers it. We will review in this article the physiology involved in its physical structure, production, and pulmonary effects.

Protein interactions with osseointegrable titanium implants

This review aims to present the mechanisms of protein interactions with titanium dental implant surfaces. Methods: the analyses were based on searches of scientific articles available in English and Portuguese in PubMed (MEDLINE), Bireme (LILACS), Scielo, Web of Science and Google Scholar. Results: titanium dental implant treatments success rates (95-98%) are mainly due to the biocompatibility of titanium oxide on the implant surface, surgical techniques adopted, good implants manufacturing processes and biomechanical knowledge of the systems. Studies in past decades has empirically developed implant surfaces with significant changes in morphologies, roughness, wettability, surface energy, chemical composition, and chemical groups density or deposited molecules. These changes promoted better protein adsorption, osteoblast adhesion, and changes in the mechanisms involved in osseointegration. Thus, the time to put the implant in function has been reduced and the success rates have increased. In the osseointegration process, at the nanoscale, there is no contact between the bone and the implant surface, but there is the formation of a protein anchorage between the periosteum and the implant with an interface formed by proteins. In all the reactions between the body and the implant surface, the activities of fibronectin and integrin are essential, since they are responsible for transmitting information to the cell for its differentiation, adhesion and mobility. Conclusion: thus, the analyses of protein-implant interactions are indispensable for a better understanding of the performance of osseointegrated dental implants

The peri-implant ligament: a scoping review

The peri-implant ligament is formed from the interface of bone tissue, through the anchoring of proteins and the surface of the dental implant. In this sense, it is relevant to understand the extent to which this ligament is structured and biomimics the periodontal ligament functions. Aim: The goal of this scoping review is to present and analyze the peri-implant ligament composition and compare the extent to which this ligament is structured and biomimics the periodontal ligament functions. Methods: This scoping review was performed according to the Joanna Briggs Institute methodology for scoping reviews and following the Preferred Reporting Items for Systematic Reviews and Meta-analyses extension for scoping review. Two independent researchers searched Pubmed, Cochrane, Embase, Virtual Health Library, Scielo, Scopus, Web of Science, Brazilian Bibliography of Dentistry, Latin American and Caribbean Literature in Health Sciences, Digital Library of Theses and Dissertations from the University of São Paulo and Portal Capes. Studies published in English, Portuguese and Spanish, over the last 21 years (2000-2021). Results: A total of 330 titles were identified and after applying inclusion and exclusion factors, 27 studies were included in this review. All proteins were identified regarding their tissue function and classified into 6 major protein groups. After that this new protein ligament was compared with the periodontal ligament regarding its function and composition. The main proteins associated with osseointegration, and thus, with the peri-implant ligament are recognized as belonging to the periodontal ligament. Conclusion: This scoping review results suggest evidence of the composition and function of the periimplant ligament. However, variations may still exist due to the existence of several modulants of the osseointegration process

Evaluation of interactions of silibinin with the proteins ALS3 and SAP5 against Candida albicans / Avaliação das interações da silibinina com as proteínas ALS3 e SAP5 contra Candida albicans

Objective: to evaluate the molecular interaction of silibinin with the targets ALS3 and SAP5. Methodology: Molecular docking protocols were conducted to analyze the binding interaction of silibinin with ALS3 and SAP5. Results: Eleven interactions of ALS3 with silibinin and four with fluconazole were found, while six interactions were observed of SAP5 with silibinin and four with fluconazole. Conclusion: Molecular docking between silibinin and ALS3 identified important interactions, but no significant interactions were observed with SAP5, even though silibinin can exhibit affinity and interactions with other SAP5 sites.

Objetivo: Avaliar a interação molecular da silibinina com os alvos ALS3 e SAP5. Metodologia: Protocolos de docking molecular foram conduzidos para analisar a interação de ligação da silibinina com ALS3 e SAP5. Resultados: Foram encontradas onze interações de ALS3 com silibinina e quatro com fluconazol, enquanto seis interações foram observadas de SAP5 com silibinina e quatro com fluconazol. Conclusão: Docking molecular entre silibinina e ALS3 identificou interações importantes, mas não foram observadas interações significativas com SAP5, embora a silibinina possa apresentar afinidade e interações com outros sítios SAP5.

Effectiveness Comparison of Polysaccharides, Proteins, and Lipids as Composite Edible Coatings on the Quality of Food Products / Comparación de la Efectividad de Polisacáridos, Proteínas y Lípidos como Recubrimientos Compuestos Comestibles en la Calidad de los Productos Alimenticios

Background: This research was motivated by the complaints of tomato farmers about their crops that quickly rotted before being sold, as well as the many research results (raw materials and methods) that edible coating films could not be applied optimally. Objectives: The research was a practical recommendation by comparing the effectiveness of raw materials (polysaccharides, proteins, and lipids) with the dipping and spray methods. Materials and methods used in the comparison process were the application of Structural Equation Modeling (SEM) with the Partial Least Square (PLS) approach. Results: Dipping has a strong effect (f2 ≥ 0.35; p<0.05), while spray had a moderate effect (f2: 0.15-0.35; p<0.05). Thus, the role of dipping as a mediator was more dominant than spray. Compared to proteins and lipids, polysaccharides had the best effectiveness (ß:0.460-0.584; f2: 0.15-0.35; p<0.05). Conclusion: the three ingredients improved the quality of tomatoes, and the dipping method was easier to apply by farmers than the spray method, which had many obstacles in its application

Antecedentes: esta investigación está motivada por las quejas de los productores de tomate sobre sus cultivos que se pudren rápidamente antes de ser vendidos, así como por los muchos resultados de la investigación (materias primas y métodos) de que las películas de recubrimiento comestibles no se pudieron aplicar de manera óptima. Objetivos: La investigación consiste en recomendaciones prácticas mediante la comparación de la eficacia de las materias primas (polisacáridos, proteínas y lípidos) con los métodos de inmersión y aspersión. Métodos: El método utilizado en el proceso de comparación es la aplicación del modelo de ecuaciones estructurales (SEM) con el enfoque de mínimos cuadrados parciales (PLS). Resultados: La inmersión tiene un efecto fuerte (f2 ≥ 0,35; p<0,05), mientras que la pulverización tiene un efecto moderado (f2: 0,15-0,35; p<0,05). Por lo tanto, el papel de la inmersión como mediador es más dominante que el del rociado. Los polisacáridos tienen la mejor eficacia (ß:0,460-0,584; f2: 0,15-0,35; p<0,05) en comparación con las proteínas y los lípidos. Conclusión: es que los tres ingredientes pueden mejorar la calidad de los tomates, y el método de inmersión es más fácil de aplicar por los agricultores que el método de aspersión, que tiene muchos obstáculos en su aplicación

Nefronoptisis: reporte de un caso pediátrico / Nephronophthisis: a pediatric case report

La nefronoptisis es una enfermedad renal quística, de herencia autosómica recesiva, causada por mutaciones en genes que codifican proteínas involucradas en la función de cilios primarios, lo que resulta en enfermedad renal y manifestaciones extrarrenales como degeneración retiniana y fibrosis hepática. Según la edad de desarrollo de enfermedad renal crónica terminal, se describen tres formas clínicas de presentación: infantil, juvenil y adolescente. El diagnóstico se realiza por una prueba genética positiva o una biopsia de riñón que demuestre cambios tubulointersticiales crónicos con un engrosamiento de las membranas basales tubulares. No existe hasta la actualidad una terapia curativa, por lo que el trasplante renal oportuno es determinante en cuanto al pronóstico. Se presenta un paciente de 13 meses de edad con poliuria de 3 meses de evolución, insuficiencia renal, anemia y elevación de transaminasas. Con hallazgos histológicos compatibles en la biopsia renal, se arribó al diagnóstico de nefronoptisis infantil, con afectación hepática

Nephronophthisis is an autosomal recessive cystic kidney disease caused by mutations in genes that encode proteins involved in the primary cilia function, resulting in kidney disease and extrarenal manifestations such as retinal degeneration and liver fibrosis. According to the age of development of end-stage chronic kidney disease, three clinical forms of presentation are described: infantile, juvenile and adolescent. Diagnosis is made by a positive genetic test, or a kidney biopsy demonstrating chronic tubulointerstitial changes with thickening of the tubular basement membranes. At the moment there is no healing therapy, so early kidney transplant is a fundamental tool to improve prognosis.We present a 13-month old male patient with polyuria, kidney failure, anemia and elevated aminotransferases over three months. With compatible histological kidney biopsy, the diagnosis of infantile nephronophthisis with liver involvement was reached.

Validación de un método analítico para cuantificación de ferritina en líquido cefalorraquídeo / Validation of an analytical method for the quantification of ferritin in cerebrospinal fluid

Resumen La ferritina es una proteína de gran tamaño que se encuentra fisiológicamente en el líquido cefalorraquídeo (LCR) en concentraciones de 2-10 ng/mL. Su elevación puede utilizarse como biomarcador en distintas condiciones patológicas. El procedimiento de validación tradicional para la medición en LCR no puede ser utilizado debido a la ausencia de controles y calibradores comerciales para esta matriz. El objetivo de este trabajo fue llevar a cabo una validación analítica de ferritina en LCR. Se realizaron ensayos de estimación de precisión y veracidad mediante el protocolo EP15-A3, linealidad por el protocolo EP6-A (ambos de la guía de la CLSI), recuperación, estabilidad, contaminación por arrastre, interferencia por hemólisis y bilirrubina y límite de detección (LoD). La ferritina en LCR en el autoanalizador DxI 800 de Beckman Coulter tuvo una performance intra e interensayo <3,7%, el ensayo denota linealidad en el intervalo de 2,1-547 ng/mL; se estableció estabilidad por un período de 5 días y la recuperación resultó ser aceptable. No se evidenció efecto de contaminación por arrastre ni interferencia por hemólisis hasta un rango entre 300-500 mg/dL de hemoglobina, ni interferencia por bilirrubina hasta una concentración de 16,0 mg/dL de bilirrubina total. El LoD fue de 0,4 ng/mL. Por medio de los ensayos realizados se logró validar la ferritina en LCR a partir de la utilización de pools de muestras, lo que pudo garantizar la confiabilidad y exactitud del método analítico.

Abstract Ferritin is a large protein physiologically present in the cerebrospinal fluid (CSF) in concentrations of 2-10 ng/mL. Its elevation can be used as a biomarker in several pathological conditions. The traditional validation procedure for measurement in CSF cannot be used due to the absence of commercial controls and calibrators for this matrix. The objective of the present study was to perform CSF ferritin analytical validation. Assays such as precision and accuracy estimation through the EP15-A3 protocol, linearity according to the EP6-A protocol (both from the CLSI guidelines), recovery, stability, carry-over, hemolysis and bilirubin interference and limit of detection (LoD) were conducted. Serum samples with different concentrations of ferritin were added to aliquots of a normal CSF pool. CSF ferritin in the Beckman Coulter DxI 800 had a <3.7% intra and inter-assay performance, the assay shows linearity in the 2.1 -547 ng/mL interval, stability was established for a 5-day period and the recovery was acceptable. There was neither carry-over effect or hemolysis interference up to a range of 300-500 mg/dL of hemoglobin, nor interference by bilirubin up to 16.0 mg/dL of total bilirubin. The LoD was 0.4 ng/mL. By means of the performed assays, CSF ferritin was validated by using sample pools, thereby ensuring the reliability and accuracy of the analytical method.

Resumo A ferritina é uma grande proteína fisiologicamente encontrada no líquido cefalorraquidiano (LCR) em concentrações de 2 a 10 ng/mL. Sua elevação pode ser usada como biomarcador em diferentes condições patológicas. O procedimento de validação tradicional para medição no LCR não pode ser usado devido à ausência de controles e calibradores comerciais para esta matriz. O objetivo deste estudo foi realizar uma validação analítica da ferritina no LCR. Foram realizados estudos de precisão e veracidade utilizando o protocolo EP15-A3, linearidade pelo protocolo EP6-A (ambos das diretrizes do CLSI), recuperação, estabilidade, contaminação transportada, interferência de hemólise e bilirrubina e limite de detecção (LoD). A ferritina no LCR no DxI 800 da Beckman Coulter teve um desempenho intra e inter-ensaio <3,7%, o ensaio denota linearidade na faixa de 2,1-547 ng/mL, a estabilidade foi estabelecida em um período de 5 dias e a recuperação foi considerado aceitável. Nenhum efeito de transporte ou interferência por hemólise foi evidenciado até um intervalo entre 300-500 mg/dL de hemoglobina, nem interferência pela bilirrubina até uma concentração de 16,0 mg/dL de bilirrubina total. O LoD foi de 0,4 ng/mL. Através dos testes realizados, a ferritina no LCR foi validada, com base no uso de pool de amostras, o que poderia garantir a confiabilidade e a acurácia do método analítico.

Fórmula artesanal a base de lactosuero: complemento alimenticio para niños preescolares / Artisanal whey-based formula: food supplement for preschool children

Resumen Se pretendió desarrollar una fórmula artesanal, a base de lactosuero, como complemento alimenticio para niños preescolares. Se realizó una investigación descriptiva ejecutada en tres fases: 1. Ensayos preliminares para la determinación del esquema tecnológico; 2. Evaluación fisicoquímica para la caracterización del producto y determinación de macronutrientes y 3. Evaluación sensorial donde se midió el nivel de agrado del producto final. Los datos obtenidos de los análisis se tabularon en cuatro repeticiones y se analizaron a través de estadísticas descriptivas de tendencia central y en frecuencias expresadas en tablas y gráficos mediante el programa estadístico SPSS versión 20.0. Se obtuvo que en el análisis proximal del requesón deshidratado, éste aportó por cada 100 gramos de producto: 480,28 kcal, 46,5% de proteínas, 22,36% de grasas y 23,26% de hidratos de carbono. La formulación final de la bebida constó de 2,9 g de requesón deshidratado, 3,6 g de arroz previamente cocido y 1,8 g de azúcar diluidos por cada onza preparada. Se determinó que es una fórmula hipocalórica-hiperproteica e isoosmolar, con una viscosidad de 275cP, un pH de 5,1 y con 0,291% de ácido láctico. La fórmula artesanal a base de lactosuero fue de agrado para 41 niños que participaron en el análisis sensorial. Se recomienda su uso en niños que se encuentren en condición de vulnerabilidad nutricional.

Abstract The main objective of this research was to develop an artisan formula based on whey as food supplement directed to preschool children. It was a descriptive study carried out in three phases: 1. Preliminary tests, for the determination of the technological scheme; 2. Physical-chemical evaluation, for the characterisation of the product and determination of nutrients and, 3. Sensory evaluation: the level of satisfaction of the final product measured. The data obtained from the analysis were tabulated in four repetitions and analysed through descriptive statistics of central tendency and in frequencies expressed in tables and graphs using the statistical program SPSS version 20.0. As a result, for each 100 grams of dehydrated cottage cheese this malnuprovides: 480.28 kcal, 46.5% protein; 22.36% fat and 23.26% carbohydrates. The final formulation of the drink consisted of 2.9 g of dehydrated cottage cheese, 3.6 g of previously cooked rice and 1.8 g of diluted sugar for each prepared ounce. It was determined as a hypocaloric-hyperproteic and isomolar formula, with a viscosity of 275cP, a pH of 5.1 and with 0.291% lactic acid. The artisan formula based on whey was liked by 41 children who participated in the sensory analysis. As a conclusion, it can be recommended as food supplement in children with nutritional vulnerability conditions.

Resumo O objetivo principal desta pesquisa foi desenvolver uma fórmula artesanal à base de soro de leite como suplemento alimentar direcionado a crianças pré-escolares. Foi realizado um estudo descritivo em três fases: 1. Ensaios preliminares, para determinação do esquema tecnológico; 2. Avaliação físico-química, para caracterização do produto e determinação de macronutrientes e 3. Avaliação sensorial: mediu-se o grau de satisfação do produto final. Os dados obtidos das análises foram tabulados em quatro repetições e analisados por meio de estatísticas descritivas de tendência central e em frequências expressas em tabelas e gráficos utilizando o programa estatístico SPSS versão 20.0. Como resultado da análise proximal, para cada 100 gramas de requeijão desidratado fornece: 480,28 kcal, 46,5% de proteína; 22,36% de gordura e 23,26% de carboidratos. A formulação final da bebida consistiu em 2,9 g de requeijão desidratado, 3,6 g de arroz previamente cozido e 1,8 g de açúcar diluído para cada onça preparada. O resultado concluiu que é uma fórmula hipocalórica-hiperproteica e isoosmolar, com viscosidade de 275cP, pH de 5,1 e com 0,291% de ácido lático. A fórmula artesanal à base de soro de leite foi apreciada por 41 crianças que participaram da análise sensorial. É recomendado seu uso em crianças que se encontrem em condições de vulnerabilidade nutricional.

Mobilization of seed storage proteins is crucial to high vigor in common bean seeds / Mobilização das proteínas de reserva é crucial para o alto vigor em sementes de feijão

Seed germination is a complex process controlled by many factors, in which physical and biochemical mechanisms are involved and the mobilization of reserves is crucial for this process to occur. Although, seed reserve mobilization is usually thought to be a post-germination process, seed reserve proteins mobilization occurs during germination. This study quantified seed proteins of bean genotypes during different hydration times, in order to understand the process of protein mobilization and whether there is relationship of this biochemical component with seed vigor. This study was conducted using seeds with different levels of vigor, genotypes with highest (13, 42, 55 and 81) and lowest (07, 23, 44, 50, IPR-88-Uirapurú and Iapar 81) physiological quality. High vigor genotypes showed greater efficiency in hydrolysis and mobilization of protein component, because they presented low globulins content in cotyledons at radicle protrusion in relation to low vigor genotypes (07, 23 and 50). The protein alpha-amylase inhibitor, observed in all genotypes, is involved with the longer time needed for radicle protrusion, according to the band intensity difference in genotypes 07, 44 and Iapar 81.

A germinação de sementes é um processo complexo controlado por muitos fatores, nos quais mecanismos físicos e bioquímicos estão envolvidos e a mobilização de reservas é decisiva para que esse processo ocorra. Embora a mobilização de reservas de sementes seja considerada um processo pós-germinativo, a mobilização das proteínas de reserva de sementes ocorre durante a germinação. Este estudo teve como objetivo quantificar as proteínas de sementes de genótipos de feijão durante os diferentes tempos de hidratação, a fim de compreender o processo de mobilização proteica e se há relação desse componente bioquímico com o vigor das sementes. Este estudo foi realizado utilizando sementes com diferentes níveis de vigor, genótipos com maior (13, 42, 55 e 81) e menor (07, 23, 44, 50, IPR-88-Uirapurú e Iapar 81) qualidade fisiológica. Os genótipos de alto vigor apresentaram maior eficiência na hidrólise e mobilização do componente proteico, pois apresentaram baixo teor de globulinas nos cotilédones na protrusão radicular em relação aos genótipos de baixo vigor (07, 23 e 50). A proteína inibidora da alfa-amilase, observada em todos os genótipos, está envolvida com o maior tempo necessário para a protrusão da radícula, de acordo com a diferença de intensidade da banda nos genótipos 07, 44 e Iapar 81.

Adecuación del aporte de proteína natural en el tratamiento de la fenilcetonuria: una revisión bibliográfica / Adequacy of natural protein intake in patients with phenylketonuria: bibliographic review

Introducción: la fenilcetonuria (PKU) es el error congénito del metabolismo de las proteínas más frecuente. El tratamiento dietético consiste en un plan de alimentación con una ingesta de proteínas naturales restringida, un sustituto proteico libre o de bajo contenido en fenilalanina (Phe) y el aporte de alimentos muy bajos en proteínas. El objetivo principal de este trabajo fue investigar si es posible aumentar la ingesta de proteína natural (PN) que se indica a los pacientes con PKU manteniendo los dosajes de Phe en sangre en rangos de seguridad. Materiales y método: se buscaron en 6 bases de datos electrónicas artículos publicados. Se identificaron un total de 154 artículos de Pub Med por intervalo de años desde 1999 a 2020. Se eligieron 15 artículos que se adaptaron a los criterios de inclusión y exclusión y respondían al objeto de estudio de esta revisión bibliográfica. Resultados: hay varios factores que pueden influenciar la estimación de la tolerancia de Phe como la severidad del fenotipo del paciente, la edad, el rango de seguridad de Phe en sangre, la prescripción de Phe y la adherencia al sustituto proteico. Si los niveles de Phe en sangre se mantienen en forma constante dentro del rango adecuado y por un período determinado, se debería considerar un incremento de la ingesta de Phe. El aumento de la ingesta de PN deberá ser realizado de manera controlada, individual y evaluando en forma constante el impacto en los dosajes de Phe en sangre. Conclusión: optimizar la ingesta de PN ofrece una mejora en la calidad de vida de pacientes con PKU, facilita la capacidad del paciente para socializar y contribuye a una mejor adherencia a la dieta(AU).

Introduction: phenylketonuria (PKU) is the most frequent inborn error of protein metabolism. The dietary treatment consists of a diet with a restricted natural protein intake, a free or low phenylalanine (Phe) protein substitute, and the intake of low protein food. The main objective of this work is to analyze if it is possible to increase the natural protein (NP) intake prescribed to PKU patients while maintaining blood Phe dosages within safe range. Materials and method: studies published were searched in 6 electronic data- basis. A total of 154 Pub Med articles were identified by range of years from 1999 to 2020. Fifteen articles which met the inclusion and exclusion criteria and responded to the objective of this bibliographic review were chosen. Results: several factors may influence Phe tolerance, such as severity of the patient´s phenotype, age, blood Phe safe range, Phe prescription and adherence to protein substitute. If Phe blood levels remain constantly within safe range and for a certain period, an increase of Phe intake should be considered. Increase of NP intake must be carried out in a controlled manner, individually and constantly evaluating blood Phe levels. Conclusion: optimizing NP intake offers the PKU patient an improvement in quality of life, facilitates the patient´s ability to socialize and contributes to a better adherence to the diet(AU).

Avaliação do perfil de microRNAs exossomais em portadores de hipercolesterolemia familial / Evaluation of the profile of exosomal microRNAs in patients with familial hypercholesterolemia

A Hipercolesterolemia Familial (HF) é uma doença hereditária do metabolismo lipídico que causa aos portadores alta incidência de aterosclerose prematura. A HF pode ser diagnosticada clínica e geneticamente, entretanto, apenas cerca de 40% podem ter confirmados pelo diagnostico molecular. Assim, outros sistemas de diagnóstico devem ser avaliados. Ultimamente devido a estabilidade em fluidos biológicos, os exossomos circulantes apresentam grande potencial, pois carreiam um número variado de compostos e são considerados veículos de intercomunicação entre os tecidos. Sabe-se que vários RNAs são carreados nos exossomos, incluindo miRNAs, lncRNA e uma variedade de proteínas. Estes componentes podem ser marcadores de diagnóstico para várias doenças inclusive a HF e suas complicações cardiovasculares. Foram utilizadas amostras de exossomos plasmáticos provenientes de 54 pacientes HF sem uso de estatina por, no mínimo, seis semanas, e 38 indivíduos normolipidêmicos para sequenciamento de miRNAs e estudo da proteômica. Os exossomos foram isolados utilizando dois métodos precipitação química e cromatográfica de exclusão de tamanho e caraterizados utilizando: dispersão de luz dinâmica, Western blotting, rastreamento de nanopartículas (NanoSight), imunomarcação e microscopia eletrônica de transmissão. Os miRNAs e proteínas foram extraídos dos exossomos e analisados por sequenciamento de nova geração e espectrometria de massa, respectivamente. Os dados clínicos, biodemográficos e laboratoriais dos pacientes HF e controles indicaram diferenças significativas esperadas entre os grupos, indicando que foram selecionados adequadamente. A caracterização físico-química dos exossomos mostrou resultados com tamanho de ˜90nm e imunorreação positiva para tetraspaninas. O resultado do sequenciamento identificou acima 2000 miRNAs. Os miR-122- 5p e miR-21-5p apresentaram expressão aumentada no grupo HF (log2FC=1,79 e log2FC=1,27, respectivamente), e o miR-122-5p pós normalização em relação ao controle manteve significativo comparados ao controle (p=0,034). A análise comparativa entre exossomos e plasma total mostrou diferença significativa, pois foram identificadas 239 proteínas (p <0,05) diferentes entre exossomos e plasma. Em exossomos, 17 proteínas foram aumentadas e 21 diminuídas em pacientes com HF em comparação com o controle (p <0,05). Destas, seis proteínas foram mais abundantes em HF e sete proteínas foram menos abundantes em exossomos de pacientes com HF em comparação com o controle. A análise de enriquecimento por bioinformática mostrou que a maior parte dessas moléculas (miRNAs e proteínas) foram relacionadas com metabolismo lipídico, dislipidemia, aterosclerose, doença arterial coronariana, adipogênese. Assim, na busca de novos alvos como potenciais biomarcadores de diagnóstico da HF, nossos resultados da análise integrativa entre os miRNAs e as proteínas exossomais abre novas frentes de pesquisa mais bem direcionadas, para a validação desses miRNAs e proteínas exossomais

Familial Hypercholesterolemia (FH) is an inherited disease of lipid metabolism that causes a high incidence of premature atherosclerosis in patients. FH can be diagnosed clinically and genetically, however, only about 40% can be confirmed by molecular diagnosis. Thus, other diagnostic systems should be evaluated. Lately, due to stability in biological fluids, circulating exosomes have great potential, as they carry a varied number of compounds and are considered vehicles of intercommunication between tissues. Several RNAs are known to be carried on exosomes, including miRNAs, lncRNA, and a variety of proteins. These components can be diagnostic markers for several diseases including FH and its cardiovascular complications. Plasma exosome samples from 54 FH patients without statin use for at least six weeks and 38 normolipidemic individuals were used for miRNA sequencing and proteomics studies. Exosomes were isolated using two methods chemical precipitation and size exclusion chromatography and characterized using: dynamic light scattering, Western blotting, nanoparticle tracking (NanoSight), immunostaining and transmission electron microscopy. MiRNAs and proteins were extracted from exosomes and analyzed by next-generation sequencing and mass spectrometry, respectively. Clinical, biodemographic and laboratory data of FH patients and controls indicated significant expected differences between the groups, indicating that they were appropriately selected. The physicochemical characterization of exosomes showed results with a size of ˜90nm and positive immunoreaction for tetraspanins. The sequencing result identified above 2000 miRNAs. miR-122-5p and miR-21-5p showed increased expression in the FH group (log2FC=1.79 and log2FC=1.27, respectively), and miR122-5p after normalization in relation to the control remained significant compared to the control (p=0.034). The comparative analysis between exosomes and total plasma showed a significant difference, as 239 different proteins (p < 0.05) were identified between exosome and plasma. In exosomes, 17 proteins were increased and 21 decreased in FH patients compared to control (p < 0.05). Of these, six proteins were more abundant in FH and seven proteins were less abundant in exosomes from patients with FH compared to the control. Bioinformatics enrichment analysis showed that most of these molecules (miRNAs and proteins) were related to lipid metabolism, dyslipidemia, atherosclerosis, coronary artery disease, adipogenesis. Thus, in the search for new targets as potential diagnostic biomarkers of FH, our results of the integrative analysis between miRNAs and exosomal proteins opens new and better-directed research fronts for the validation of these miRNAs and exosomal proteins

Cristalogénesis biológica y cristalografía en la elucidación de la estructura tridimensional de las proteínas. Revisión narrativa / Protein tridimensional structure by biological crystallogenesis and crystallography. Narrative review

La obtención de información estructural tridimensional de una proteína es de suma importancia en campos tan variados como la bioquímica funcional, las ciencias de materiales o biomédicas. Siendo actualmente la difracción de rayos X de monocristal el estándar de oro para la consecución de este objetivo, la obtención de dicho monocristal sigue siendo un cuello de botella desde el punto de vista práctico, y poco entendido desde el punto de vista teórico. En este artículo se revisa desde la perspectiva estructural de la proteína la forma en que los rayos X permiten obtener la información estructural y las condiciones fisicoquímicas que permiten la formación de un cristal adecuado para estos experimentos.

Obtaining three-dimensional structural information of a protein is of utmost importance in various fields such as functional biochemistry, materials science, or biomedical sciences. Even though single crystal X-ray diffraction is currently the gold standard for this purpose, growing said single crystal is still a bottleneck from a practical viewpoint, and not fully understood from a theoretical point of view. In this article, we review, from a protein structure perspective, the way X-rays provide structural information, and the physicochemical conditions that promote the formation of an adequate crystal for these experiments.

Amiloidosis AL: conceptos actuales / AL amyloidosis: current concepts

Las amiloidosis sistémicas constituyen un grupo de enfermedades con diversas etiologías, caracterizadas por la síntesis de proteínas con plegado defectuoso, capaces de agregarse y depositarse en el medio extracelular de diferentes órganos y tejidos, alterando su estructura y función. Se conocen más de 14 formas de amiloidosis sistémica, de las cuales la más frecuente es la amiloidosis AL, objeto de esta revisión, en la que las proteínas precursoras son cadenas ligeras de inmunoglobulina inestables, secretadas por un clon de células plasmáticas o, con menor frecuencia, por un linfoma linfoplasmocítico o de células del manto. La amiloidosis AL puede llevar a una amplia gama de manifestaciones clínicas y compromiso de órganos, como el corazón y el riñón. El reconocimiento temprano de la enfermedad y el diagnóstico oportuno son determinantes para mejorar la supervivencia de los pacientes. El tratamiento deberá ser individualizado de acuerdo con la condición de cada paciente, lo que hace necesaria una correcta clasificación de los individuos según su pronóstico. La terapia dirigida a la amiloidosis está enfocada esencialmente en disminuir el compromiso orgánico, y por ende, prolongar la supervivencia con mejoría en los síntomas. En esta revisión se discutirán aspectos importantes de la fisiopatología, epidemiología, manifestaciones clínicas, diagnóstico y tratamiento de la amiloidosis AL

Systemic amyloidosis constitutes a group of diseases with diverse etiologies characterized by the synthesis of proteins with defective folding, capable of aggregating and depositing in the extracellular matrix of different organs and tissues, altering their structure and function. More than 14 forms of systemic amyloidosis are known, of which the most frequent is AL amyloidosis, the subject of this review, in which the precursor proteins are unstable immunoglobulin light chains, secreted by a clone of plasma cells or, to a lesser extent, often due to lymphoplasmacytic or mantle cell lymphoma. AL amyloidosis can lead to a wide range of clinical manifestations and organ involvement, such as the heart and kidney. Early recognition of the disease and timely diagnosis are crucial to improve patient survival. Treatment should be individualized according to the condition of each patient, which requires a properly classification of individuals according to their prognosis. Amyloidosis-targeted therapy is essentially focused on reducing organ involvement, and therefore prolonging survival with improvement in symptoms. In this review, important aspects of the pathophysiology, epidemiology, clinical manifestations, diagnosis, and treatment of AL amyloidosis are discussed

Semisolid formulations based on solid-in-oil-in-water systems containing proteins

Abstract In recent years, improvements have been made, through biotechnological processes, in the production and development of peptides capable of increasing collagen and elastin synthesis for anti-aging skin care. However, proteins have many limitations due to their structural, chemical and physical fragility to external aggressions, which may cause conformational changes, leading to loss of biological activity. Therefore, it is important to create delivery systems that protect these biomolecules from damage, allowing them to reach their target. This work aimed to develop a system able to carry bovine serum albumin (BSA), used as a model of a protein, and to incorporate this system in a semisolid formulation suitable for skin application. A microemulgel based on a solid-in-oil-in-water (S/O/W) microemulsion was prepared. Firstly, the association efficiency (AE) of lyophilized BSA-sucrose ester complex and the size of S/O nanodispersion were assessed; then, the characterization and stability evaluation of the final semisolid formulation through evaluation of pH, texture and rheological behavior were performed. The average value of AE was 54.74% ± 2.17. It was possible to develop an S/O/W microemulsion, which allowed the subsequent development of an S/O/W microemulgel that assured suitable pH, texture and rheological characteristics for skin application.

Design of Inhibitory Peptides of the Interaction between the E2 Protein of the Hepatitis C Virus and the CD81 and CD209 receptors / Diseño de péptidos inhibitorios de la interacción entre la proteína E2 del virus de Hepatitis C y los receptores CD81 y CD209

Abstract Infection through the Hepatitis C virus does not have a vaccine and treatment with pegylated interferon and ribavirin can fail; which is why it may cause chronic infection and, consequently, could develop liver failure or hepatocellular carcinoma. It has been described that virus-cell recognition occurs between the E2 viral envelope protein and diverse cell receptors, with this interaction being critical in viral infection. which is why the study sought to identify inhibitory peptides of the interaction between viral E2 protein and the CD81 and CD209 receptors. Methodology: Through the RCSB protein database, crystals from the CD81 and CD209 receptors were selected, CD81/E2-HCV, CD209/E2-HCV complexes were carried out by SWISS-MODEL to generate inhibitory peptides of protein interaction through the Rosetta web server, this interaction was validated through ClusPro and finally, determined the theoretical physicochemical and cytotoxic properties of these peptides. Results: two peptides were obtained, without predicted toxicity, with a theoretical capacity of blocking the protein interaction between the E2 protein of the virus and CD81 and CD209.

Resumen La infección por el virus de la hepatitis C, no cuenta con vacuna y el tratamiento con interferón pegilado y ribavirina puede fallar; por lo que puede causar infec ción crónica y como consecuencia podría desarrollarse falla hepática o carcinoma hepatocelular. Se ha descrito que el reconocimiento virus-célula, se da entre la proteína de envoltura viral E2 y diversos receptores celulares, siendo esta interacción crítica en la infección viral. Razón por la cual este estudio buscó identificar péptidos inhibidores de la interacción entre la proteína E2 viral y los receptores CD81 y CD209. Metodología: A través de la base de datos de proteínas RCSB, se seleccionaron cristales de los receptores CD81 y CD209, se realizaron complejos CD81/E2-HCV, CD209/E2-HCV para generar péptidos inhibidores de interacción proteica a través del servidor web Rosetta, esta interacción fue validada a través de ClusPro y finalmente se evaluaron las propiedades fisicoquímicas y citotóxicas teóricas para estos péptidos. Resultados: se obtuvo dos péptidos, sin toxicidad predicha, con capacidad teórica de bloquear la interacción proteica entre la proteína E2 del virus y CD81 y CD209.

SFRP1, Posible biomarcador en la progresión o regresión de lesiones de cérvix asociado al Virus del Papiloma Humano / SFRP1, Possible Biomarker in the Progression or Regression of Cervical Pre-neoplastic Lesions Associated with Human Papilloma Virus

Resumen Objetivo: Comparar la expresión de mRNA y proteínas de SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 y HAND1 en pacientes con lesión intra-epitelial cervical de bajo y alto grado, con posterior progresión o regresión. Material y Método: Se realizó análisis de expresión de genes mediante RT-PCR y análisis de expresión de proteínas por inmunohistoquímica. El análisis estadís tico fue realizado con las pruebas: Wilcoxon, coeficiente de correlación de Spearman e índice de concordancia. Las muestras fueron pareadas en momento 1 y momento 2. Resultados: SFRP1 mostró tendencia de mayor expresión de mRNA en lesión intra-epitelial de bajo grado (momento 2) Vs. alto grado (momento 1). La expresión de proteínas por inmunohistoquímica de SFRP1 en casos de progresión (83,3 %) mostró disminución en su graduación (p = 0,0313*); los demás genes en estudio no tuvieron cambios estadísticamente significativos. Discusión: SFRP1 mostró comportamiento ajustado a resultados de estudios previos donde se encontró hipermetilado en lesiones intra-epiteliales de alto grado; su subexpresión por hipermetilación se reportó en otros canceres, proceso que colabora con su silenciamiento y transición epitelial-mesenquimatosa del cáncer de cuello uterino. Conclusiones. SFRP1 es potencial biomarcador en lesiones preneoplásicas del cuello uterino asociadas al virus de papiloma humano.

Abstract Objective. The aim of this work was to compare the expression of mRNA and proteins of SFRP1, PTPRN, CDO1, EDNRB, CDX2, EPB41L3 and HAND1 in patients with low and high grade cervical intraepithelial lesion, with subsequent progression or regression. Material and Methods: Gene expression analysis was conducted through RT-PCR and protein expression analysis was performed by immunohistochemistry. The statistics analysis were Wilcoxon test, Spearman's correlation coefficient and concordance index. The samples were paired during moment 1 (initial patient diag nosis) and moment 2 (follow-up histological diagnosis). Results: SFRP1 showed a trend of higher mRNA expression in low-grade intra-epithelial lesions (moment 2) Vs. high-grade (moment 1). The expression of proteins by immunohistochemistry of SFRP1 in progression cases (83.3%) showed a decrease in its graduation (p = 0.0313*); the other genes under study had no statistically significant. Discussion: SFRP1 showed a biological behavior adjusted to the results of previous studies where hypermethylation was found in high-grade intra-epithelial lesions; its subexpression by hypermethylation has been reported in other cancers, a process that collaborates with its silencing and epithelial-mesenchymal tran sition of cervical. Conclusions. SFRP1 is a potential biomarker in preneoplastic lesions of the cervix associated with human papillomavirus.