Proteína morfogénica ósea en cirugía maxilofacial. Revisión bibliográfica / Bone morphogenic protein in maxillofacial surgery. Bibliographic review

El propósito de esta revisión es presentar una visión concreta de las proteínas morfogenéticas óseas, su potencial de inducir osteogénesis y la aplicación en los procesos regenerativos. Las investigaciones acerca de los iniciadores moleculares de diferenciación ósea y cartilaginosa han identificado un grupo entero de proteínas morfogenéticas óseas que ejercen efecto regulador. La proteína morfogenética ósea (BMP) es endógena, presenta propiedades osteoinductivas, osteoconductivas y osteogénicas, y ha mostrado efectos significativos en la promoción de la formación ósea, por lo cual es una buena alternativa en reconstrucción. El uso de BMP se ha descrito en la reconstrucción de los defectos óseos de origen traumático y patológico, incluyendo: fisura nasoalveolar, aumento del reborde alveolar, elevación del seno maxilar, injerto de alvéolo postextracción y cirugía peri-implantaria.

The purpose of this review is to present a concrete vision about bone morphogenetic proteins, their potential in the induction of osteogenesis and their application in regenerative processes. Research on the molecular primers of bone and cartilage differentiation has identified an entire group of bone morphogenetic proteins that exert a regulatory effect. The bone morphogenetic protein (BMP) is an endogenous protein, has osteoinductive, osteoconductive and osteogenic properties, has shown significant effects in the promotion of bone formation, being a good alternative in reconstruction. The use of BMP has been described in the reconstruction of bone defects of traumatic and pathological origin, including nasoalveolar fissure, increased alveolar ridge, maxillary sinus elevation, post-extraction alveolar graft, and perimplant surgery

Avances en la investigación del virus dengue en Colombia: papel de los microARNs celulares en la respuesta anti-dengue virus / Advances in dengue virus research in Colombia: the role of cellular microRNAs as an anti-dengue virus response

Dengue is one of the most important mosquito-borne diseases, and its incidence has increased at an alarming rate in recent years, becoming a real public health problem. Currently, there is no vaccine or medication or proper treatment for dengue control. Considering this situation, it is necessary to prioritize the search for new alternatives and strategies for dengue prevention and control, in order to reduce not only the economic burden of endemic countries, but also to improve the quality of life of patients. In this regard, a brief reflection on some aspects related to the search for new alternatives in Colombia is presented. This is focused on the use of microRNAs, which could be a new strategy with great therapeutic potential.

El dengue es una de las enfermedades más importantes transmitidas por mosquitos y su incidencia ha aumentado a un ritmo alarmante en los últimos años, al punto que se ha convertido en un verdadero problema de salud pública. Actualmente no existe ni vacuna, ni un medicamento o tratamiento adecuado para el control del dengue. Con dichos antecedentes, es necesario priorizar en la búsqueda de nuevas alternativas o estrategias de control y prevención del dengue con miras a disminuir no sólo la carga económica de los países endémicos, sino también a mejorar la calidad de vida de los pacientes. En este sentido, se presenta una breve reflexión sobre algunos aspectos relacionados con la búsqueda de nuevas alternativas en Colombia, enfocadas en el uso de los microARNs, que podrían constituir una nueva estrategia con un gran potencial terapéutico, dado que tendrían el potencial de contrarrestar algunas infecciones virales crónicas.

Mir-190b negatively contributes to the Trypanosoma cruzi- infected cell survival by repressing PTEN protein expression

Chagas disease, which is caused by the intracellular protozoanTrypanosoma cruzi, is a serious health problem in Latin America. The heart is one of the major organs affected by this parasitic infection. The pathogenesis of tissue remodelling, particularly regarding cardiomyocyte behaviour after parasite infection, and the molecular mechanisms that occur immediately following parasite entry into host cells are not yet completely understood. Previous studies have reported that the establishment of parasitism is connected to the activation of the phosphatidylinositol-3 kinase (PI3K), which controls important steps in cellular metabolism by regulating the production of the second messenger phosphatidylinositol-3,4,5-trisphosphate. Particularly, the tumour suppressor PTEN is a negative regulator of PI3K signalling. However, mechanistic details of the modulatory activity of PTEN on Chagas disease have not been elucidated. To address this question, H9c2 cells were infected with T. cruzi Berenice 62 strain and the expression of a specific set of microRNAs (miRNAs) were investigated. Our cellular model demonstrated that miRNA-190b is correlated to the decrease of cellular viability rates by negatively modulating PTEN protein expression in T. cruzi-infected cells.

Serum insulin-like growth factor-1 levels in females and males in different cervical vertebral maturation stages

OBJECTIVE: The aim of this cross sectional study was to assess serum insulin-like growth factor-1 (IGF-1) levels in female and male subjects at various cervical vertebral maturation (CVM) stages. MATERIAL AND METHODS: The study sample consisted of 60 subjects, 30 females and 30 males, in the age range of 8-23 years. For all subjects, serum IGF-1 level was estimated from blood samples by means of chemiluminescence immunoassay (CLIA). CVM was assessed on lateral cephalograms using the method described by Baccetti. Serum IGF-1 level and cervical staging data of 30 female subjects were included and taken from records of a previous study. Data were analyzed by Kruska-Wallis and Mann Whitney test. Bonferroni correction was carried out and alpha value was set at 0.003. RESULTS: Peak value of serum IGF-1 was observed in cervical stages CS3 in females and CS4 in males. Differences between males and females were observed in mean values of IGF-1 at stages CS3, 4 and 5. The highest mean IGF-1 levels in males was observed in CS4 followed by CS5 and third highest in CS3; whereas in females the highest mean IGF-1 levelswas observed in CS3 followed by CS4 and third highest in CS5. Trends of IGF-1 in relation to the cervical stages also differed between males and females. The greatest mean serum IGF-1 value for both sexes was comparable, for females (397 ng/ml) values were slightly higher than in males (394.8 ng/ml). CONCLUSIONS: Males and females showed differences in IGF-1 trends and levels at different cervical stages. .

OBJETIVO: o objetivo do presente estudo transversal foi avaliar os níveis do fator de crescimento semelhante à insulina-1 (IGF-1 sérico) em pacientes de ambos os sexos e em diferentes estágios de maturação das vértebras cervicais (MVC). MÉTODOS: a amostra consistiu de 60 pacientes, sendo 30 do sexo masculino e 30 do sexo feminino, com idades entre 8 e 23 anos. Amostras de sangue foram colhidas de todos os pacientes, cujos níveis de IGF-1 sérico foram avaliados por meio do método de imunoensaio quimioluminescente (CLIA). O estágio de MVC foi avaliado por meio de radiografias cefalométricas de perfil por meio do método descrito por Baccetti. O nível de IGF-1 sérico e o estágio de maturação das vertebras cervicais de 30 pacientes do sexo feminino foram avaliados e os dados retirados dos registros de um estudo prévio. Os dados foram submetidos aos testes de Kruskal-Wallis e de Mann-Whitney. A correção de Bonferroni foi calculada e o valor de alfa foi de 0,003. RESULTADOS: o valor de pico do IGF-1 sérico foi encontrado no estágio CS3, para mulheres, e CS4, para homens. Foram encontradas diferenças entre as médias dos valores de IGF-1 entre homens e mulheres nos estágios CS3, 4 e 5. O valor médio mais alto para os níveis de IGF-1 nos homens foi observado no estágio CS4, seguido do estágio CS5 e CS3. Nas mulheres, o valor médio mais alto foi observado em CS3, seguido do estágio CS4 e CS5. Diferenças também foram encontradas quanto à curva do IGF-1, em relação ao estágio de maturação das vértebras cervicais nos pacientes de ambos os sexos. O valor médio de IGF-1 sérico mais alto foi comparado. As pacientes do sexo feminino apresentaram valores ligeiramente mais altos (397ng/ml) em comparação aos pacientes do sexo masculino (394.8ng/ml). CONCLUSÕES: homens e mulheres apresentam valores de IGF-1 diferentes em estágios de maturação das vértebras cervicais diferentes. .

Association of junk food consumption with high blood pressure and obesity in Iranian children and adolescents: the CASPIAN-IV Study / Associação entre o consumo de junk food e a pressão arterial alta e obesidade em crianças e adolescentes iranianos: o Estudo Caspian-IV

OBJECTIVE: This study aimed to evaluate the association of junk food consumption with hypertension and obesity in a national sample of Iranian children and adolescents. METHODS: This nationwide study was conducted in 2011-2012 among 14,880 students, aged 6-18 years, selected by cluster sampling from 30 provinces. Weight, height, waist circumference (WC), hip circumference (HC), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR), as well as systolic and diastolic blood pressure (BP) were measured. Junk food was divided into four categories, including salty snacks, sweets, sweetened beverages, and fast food. Subjects reported how many times they had consumed each item (daily, weekly, and seldom). RESULTS: The intake of sweets was significantly associated with anthropometric indices and BP levels. Moreover, a significant association was found between fast food consumption, BP levels, and anthropometric indices (except for WHtR and WHR). Sweet beverages consumption was significantly associated with anthropometric indices; however, the consumption of salty snacks was only significantly associated with height, HC, and WHR. The risk of general obesity (OR: 0.75, 95% CI: 0.65-0.87) and abdominal obesity (OR: 0.81, 95% CI: 0.72-0.92) among participants who seldom consumed sweets was less than those who consumed daily. Also, the risk of general obesity (OR: 0.85, 95% CI: 0.74-0.97) among students that seldom consumed sweetened beverages was less than subjects who consumed them on a daily basis. CONCLUSION: It was found that junk food consumption increased the risk of both general and abdominal obesity; therefore, consumption of junk food should be reduced via restricting TV advertisements and increasing taxes on junk foods. .

OBJETIVO: Avaliar a associação entre o consumo de junk food e a hipertensão e obesidade em uma amostra nacional de crianças e adolescentes iranianos. MÉTODOS: Este estudo nacional foi feito entre 2011 e 2012 com 14.880 estudantes com seis-18 anos, selecionados por amostra em bloco em 30 províncias. Foram medidos o peso, a estatura, a circunferência da cintura (CC), a circunferência do quadril (CQ), a razão cintura/quadril (RCQ), a razão cintura/estatura (RCE) e a pressão arterial sistólica e diastólica (PAS e PAD). A junk food foi dividida em quatro categorias, incluindo lanches salgados, doces, bebidas açucaradas e fast food. Os indivíduos relataram quantas vezes consumiam cada um dos itens (diariamente, semanalmente, raramente). RESULTADOS: O consumo de doces foi associado significativamente aos índices antropométricos e níveis de pressão arterial (PA). Além disso, havia uma associação significativa entre o consumo de fast food e os níveis de PA e os índices antropométricos (exceto RCE e RCQ). O consumo de bebidas açucaradas foi associado significativamente aos índices antropométricos, porém o consumo de lanches salgados foi associado significativamente apenas a estatura, CQ e RCQ. O risco de obesidade geral (RC: 0,75, IC de 95%: 0,65-0,87) e obesidade abdominal (RC: 0,81, IC de 95%: 0,72-0,92) entre participantes que raramente consumiam doces era menor do que naqueles que os consumiam diariamente. Além disso, o risco de obesidade geral (RC: 0,85; IC de 95%: 0,74-0,97) entre estudantes que raramente consumiam bebidas açucaradas era menor do que entre indivíduos que os consumiam diariamente. CONCLUSÃO: Constatamos que o consumo de junk food aumentou o risco de obesidade geral e abdominal; portanto, o consumo de junk food deve ser reduzido por meio da restrição de comerciais de TV e do aumento de impostos sobre esse tipo de alimento. .

Traducción independiente de la estructura 5´cap del ARN genómico del virus dengue / 5´cap-independent translation of dengue virus genomic RNA

Objetivos. Analizar la participación de la caperuza metil-guanosín-trifosfato (5´cap) y de la región inicial del ARN genómico del virus dengue serotipo 2 (DENV-2) genotipo Americano en la traducción, utilizando un sistema libre de células obtenido de placenta humana. Materiales y métodos. Se preparó el plásmido recombinante pTZ18R-D2 conteniendo el ADN que codifica la 5´UTR y los primeros 201 nucleótidos de la cápside viral. Este plásmido se utilizó para transcribir el ARN correspondiente (ARN-D2), sin la 5´cap. El ARN-D2 fue traducido en un sistema constituido por la fracción posmitocondrial (S-30) de placenta humana y se evaluó la incorporación de [14C] aminoácidos en presencia del ARN-D2 y en su ausencia (control). Se diseñaron siete oligonucleótidos antisentido (OAs1-7) dirigidos contra secuencias de las estructuras SLA, SLB y cHP del ARN-D2 y se analizó el efecto de los mismos sobre la traducción ARN-D2. Resultados.El ARN-D2 produjo un incremento significativo (p<0,001) en la incorporación de [14C] aminoácidos, con estimulación del 75% de la actividad traduccional respecto al control. El análisis de los productos de traducción mostró un pico de incorporación correspondiente a péptidos con peso molecular aparente cercano al esperado (7,746 kDa). El OAs5, complementario a una secuencia de la estructura SLB del ARN-D2, inhibió completamente la traducción. Conclusiones. El ARN-D2 fue traducido de manera específica y eficiente, bajo condiciones semejantes a las intracelulares en humanos, por un mecanismo alternativo independiente de la 5´cap, que involucraría a la estructura SLB. Este mecanismo podría considerarse como blanco en el desarrollo de terapias antisentido para inhibir la reproducción del virus.

Objetives. To analyze the involvement of methyl guanosine triphosphate cap (5Æcap) and the start site of the genomic RNA of Dengue virus serotype 2 (DENV-2) American genotype in translation, using a cell-free system prepared from human placenta. Materials and methods. The recombinant plasmid pTZ18R-D2 was prepared containing DNA encoding the 5ÆUTR and the first 201 nucleotides of the viral capsid. This plasmid was used to transcribe the corresponding RNA (RNA-D2) without the 5Æ cap. The RNA-D2 was translated in a system consisting of the postmitochondrial fraction (S-30) from human placenta and the incorporation of [14C] aminoacids in the presence of RNA-D2 and in its absence (control) was evaluated. Seven antisenseoligonucleotides (OAs1-7) directed against sequences of the SLA, SLB and CHP structures of RNA-D2 were designed and the effect thereof on RNA-D2 translation was analyzed. Results.The RNA-D2 produced a significant increase (p<0.001) in the incorporation of [14C] amino acids, with 75% stimulation of translational activity compared to the control. Analysis of the translation products showed peak incorporation corresponding to peptides with apparent molecular weight close to the expected (7.746 kDa).The OAs5, complementary to a sequence of SLB structure of RNA-D2, completely inhibited translation. Conclusions. The RNA-D2 was translated specifically and efficiently under conditions similar to human intracellular conditions, by an alternative 5Æ cap-independent mechanism, which would involve the SLB structure. This mechanism might be seen as an aim in the development of antisense therapies to inhibit virus replication.

Natural transovarial transmission of dengue virus 4 in Aedes aegypti from Cuiabá, State of Mato Grosso, Brazil

INTRODUCTION: Dengue is the most prevalent arboviral disease in tropical areas. In Mato Grosso, outbreaks are reported every year, but studies on dengue in this state are scarce. METHODS: Natural transovarial infection of Aedes aegypti by a flavivirus was investigated in the Jardim Industriário neighborhood of Cuiabá, Mato Grosso. Eggs were collected with ovitraps during the dry, intermediate, and rainy seasons of 2012. After the eggs hatched and the larvae developed to adulthood, mosquitoes (n = 758) were identified and allocated to pools of 1-10 specimens according to the collection location, sex, and climatic period. After RNA extraction, multiplex semi-nested RT-PCR was performed to detect the four dengue virus (DENV) serotypes, yellow fever virus, West Nile virus and Saint Louis encephalitis virus. RESULTS: DENV-4 was the only flavivirus detected, and it was found in 8/50 pools (16.0%). Three of the positive pools contained females, and five contained males. Their nucleotide sequences presented 96-100% similarity with DENV-4 genotype II strains from Manaus, Amazonas. The minimum infection rate was 10.5 per 1000 specimens, and the maximum likelihood estimator of the infection rate was 11.6 (95% confidence interval: 4.8; 23.3). CONCLUSIONS: This study provides the first evidence of natural transovarial infection by DENV-4 in Ae. Aegypti in Mato Grosso, suggesting that this type of infection might serve as a mechanism of virus maintenance during interepidemic periods in Cuiabá, a city where dengue epidemics are reported every year. These results emphasize the need for efficient vector population control measures to prevent arbovirus outbreaks in the state. .

Identificação de motivos relevantes para a função do fator de iniciação da tradução EIF4E3 de Leishmania sp / Identification of relevant motifs to translation initiation factor EIF4E3 function in Leishmania sp

Os tripanossomatídeos são organismos caracterizados pelo controle póstranscricional da expressão gênica, principalmente em nível de tradução. Na tradução em eucariotos, tem grande destaque o complexo eIF4F, sendo um de seus principais componentes o fator de iniciação da tradução eIF4E. Já foram descritos em tripanossomatídeos seis homólogos para o eIF4E, nomeados EIF4E1 a 6. Em um estudo com Leishmania amazonensis, focado no EIF4E3, percebeu-se que seu perfil de expressão se alterava rapidamente numa curva de crescimento, com este apresentando ao menos duas bandas. As mudanças observadas sugeriam modificações pós-traducionais do tipo fosforilação, algo posteriormente confirmado. Analisando-se a sequência do EIF4E3 de Leishmania, foi possível identificar a presença de possíveis sítios de fosforilação e de ligação a parceiros funcionais como homólogos do eIF4G, outro componente do complexo eIF4F, e da proteína de ligação á cauda poli-A (PABP). No presente estudo foi analisado o perfil de expressão e a capacidade de ligação a parceiros funcionais do EIF4E3 de Leishmania superexpresso em células transfectadas e no qual foram introduzidas mutações em motivos específicos. Os resultados mostraram um perfil de expressão de ao menos três bandas para o EIF4E3 de L. amazonensis e duas para L. infantum, com o sítio S75, presente apenas na primeira, sendo o responsável por esta diferença...

Hepatic inducible nitric oxide synthase expression increases upon exposure to hypergravity

Stimulation by a number of conditions, including infection, cytokines, mechanical injury, and hypoxia, can upregulate inducible nitric oxide synthase (iNOS) in hepatocytes. We observed that exposure to hypergravity significantly upregulated the transcription of the hepatic iNOS gene. The aim of this study was to confirm our preliminary data, and to further investigate the distribution of the iNOS protein in the livers of mice exposed to hypergravity. ICR mice were exposed to +3 Gz for 1 h. We investigated the time course of change in the iNOS expression. Hepatic iNOS mRNA expression progressively increased in centrifuged mice from 0 to 12 h, and then decreased rapidly by 18 h. iNOS mRNA levels in the livers of centrifuged mice was significantly higher at 3, 6, and 12 h than in uncentrifuged control mice. The pattern of iNOS protein expression paralleled that of the mRNA expression. At 0 and 1 h, weak cytoplasmic iNOS immunoreactivity was found in some hepatocytes surrounding terminal hepatic venules. It was noted that at 6 h there was an increase in the number of perivenular hepatocytes with moderate to strong cytoplasmic immunoreactivity. The number of iNOS-positive hepatocytes was maximally increased at 12 h. The majority of positively stained cells showed a strong intensity of iNOS expression. The expression levels of iNOS mRNA and protein were significantly increased in the livers of mice exposed to hypergravity. These results suggest that exposure to hypergravity significantly upregulates iNOS at both transcriptional and translational levels.

Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA / Clonagem, expressão e caracterização de proteína SeM de Streptococcus equi subsp. equi e avaliação de sua utilização como antígeno em um ELISA indireto

Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd...

A adenite equina é uma enfermidade economicamente importante de equinos, causada por Streptococcus equi subsp. equi. Seu diagnóstico pode ser confirmado de forma direta, por meio de isolamento bacteriano e de PCR, ou de forma indireta, por meio de ELISA, método baseado na detecção de anticorpos séricos. O objetivo deste estudo foi clonar, expressar e caracterizar a proteína SeM de Streptococcus equi subsp. equi, avaliar sua utilização como antígeno em um ELISA indireto e determinar a capacidade do teste de distinguir soros de animais negativos, vacinados e positivos. Para tal, foi inicialmente realizada a clonagem do gene que codifica para a proteína SeM e sua expressão em Escherichia coli. Posteriormente, a proteína produzida foi caracterizada e utilizada como antígeno em um teste de ELISA indireto. Para avaliação do teste, foram utilizadas amostras de soro de 40 potros negativos, de 46 equinos vacinados com uma vacina comercial contra adenite equina e de 46 equinos com diagnóstico da doença. O teste demonstrou alta sensibilidade e especificidade, permitindo discriminar entre soros negativos e positivos, positivos e de animais vacinados, e negativos e de animais vacinados. Assim, conclui-se que a proteína rSeM produzida pode ser usada como antígeno para o diagnóstico da enfermidade e que o ELISA descrito pode ser útil para avaliar o estado imunológico do rebanho...

Determinantes e forças seletivas na evolução das proteínas / Determinants and selective forces in the evolution of proteins

A análise de grandes quantidades de dados aproveitando o poder computacional de ferramentas open sourcel que estão disponíveis na internet é o que veio a conhecer-se como quarto paradigma da investigação científica. Em muitas áreas do conhecimento como a Astronomia, a Física e Geologia, a experimentação, o desenvolvimento teórico e o poder computacional (os três primeiros paradigmas) têm dado lugar à análise rotineira de grandes quantidades de dados e o desenvolvimento de novos métodos, conceitos e teorias que permitam interpretar a informação gerada por novas tecnologias. No campo da biologia, esta mudança nos paradigmas da investigação científica supõe um desafio na hora de encarar uma questão biológica; mas, em contrapartida, ela oferece a oportunidade de validar teorias clássicas e ou testar hipóteses novas. Precisamente neste contexto, a presente tese aborda duas questões pertinentes ao campo da biologia evolutiva: Quais são os fatores que determinam a evolução de uma proteína? e Qual é a natureza da seleção cinética tradicional? Estas perguntas são, em principio, relevantes no âmbito teórico; por outro lado, sua compreensão, implicações e perspectivas têm também espaço importante na área experimental...

Proteomic analysis of the regenerating liver following 2/3 partial hepatectomy in rats

BACKGROUND: Liver regeneration (LR) after 2/3 partial hepatectomy (PH) is one of the most studied models of cell, organ, and tissue regeneration. Although the transcriptional profile analysis of regenerating liver has been carried out by many reserachers, the dynamic protein expression profile during LR has been rarely reported up to date. Therefore, this study aims to detect the global proteomic profile of the regenerating rat liver following 2/3 hepatectomy, thereby gaining some insights into hepatic regeneration mechanism. RESULTS: Protein samples extracted from the sham-operated and the regenerating rat livers at 6, 12, 24, 72, 120 and 168 h after PH were separated by IEF/SDS-PAGE and then analyzed by MALDI-TOF/TOF mass spectrometry. Compared to sham-operated groups, there were totally 220 differentially expressed proteins (including 156 up-regulated, 62 down-regulated, and 2 up/down-regulated ones) identified in the regenerating rat livers, and most of them have not been previously related to liver regeneration. According to the expression pattern analysis combined with gene functional analysis, it showed that lipid and carbohydrate metabolism were enhanced at the early phase of LR and continue throughout the regeneration process. Ingenuity Pathway Analysis indicated that YWHAE protein (one of members of the 14-3-3 protein family) was located at the center of pathway networks at all the timepoints after 2/3 hepatectomy under our experimental conditions, maybe suggesting a central role of this protein in regulating liver regeneration. Additionally, we also revealed the role of Cdc42 (cell division cycle 42) in the termination of LR. CONCLUSIONS: For the first time, our proteomic analysis suggested an important role of YWHAE and pathway mediated by this protein in liver regeneration, which might be helpful in expanding our understanding of LR amd unraveling the mechanisms of LR.

Biochemical adaptations of mammalian hibernation: exploring squirrels as a perspective model for naturally induced reversible insulin resistance

An important disease among human metabolic disorders is type 2 diabetes mellitus. This disorder involves multiple physiological defects that result from high blood glucose content and eventually lead to the onset of insulin resistance. The combination of insulin resistance, increased glucose production, and decreased insulin secretion creates a diabetic metabolic environment that leads to a lifetime of management. Appropriate models are critical for the success of research. As such, a unique model providing insight into the mechanisms of reversible insulin resistance is mammalian hibernation. Hibernators, such as ground squirrels and bats, are excellent examples of animals exhibiting reversible insulin resistance, for which a rapid increase in body weight is required prior to entry into dormancy. Hibernator studies have shown differential regulation of specific molecular pathways involved in reversible resistance to insulin. The present review focuses on this growing area of research and the molecular mechanisms that regulate glucose homeostasis, and explores the roles of the Akt signaling pathway during hibernation. Here, we propose a link between hibernation, a well-documented response to periods of environmental stress, and reversible insulin resistance, potentially facilitated by key alterations in the Akt signaling network, PPAR-γ/PGC-1α regulation, and non-coding RNA expression. Coincidentally, many of the same pathways are frequently found to be dysregulated during insulin resistance in human type 2 diabetes. Hence, the molecular networks that may regulate reversible insulin resistance in hibernating mammals represent a novel approach by providing insight into medical treatment of insulin resistance in humans.

Os sítios adicionais de trans-splicing na 5' UTR dos genes trans-sialidase de Trypanosoma cruzi e avaliação de seus efeitos sobre a tradução / Additional trans-splicing sites in the 5 'UTR of the trans-sialidase gene Trypanosoma cruzi and evaluation of their effects on translation

A regulação gênica em tripanossomatídeos é policistrônica e ocorre de maneira pós-transcricional. [...] Para verificar se UTR de tamanhos diferentes possuem impacto na transcrição e, por conseguinte, na tradução de proteínas, selecionou-se genes da família de trans-sialidases, devido à importância destas no processo de invasão celular. A metodologia envolveu a extração de RNA total de T. cruzi CL-Brener, seguido da síntese de cDNA, PCR, clonagem dos produtos amplificados e seqüenciamento por Sanger e pelo uso do 454 Junior (Next Generation Sequencing – NSG). Como as trans-sialidases correspondem a uma família gênica de cópias múltiplas, usou-se a estratégia de sequenciamento de alto-desempenho na tentativa de cobrir o maior número possível de genes desta família. Para isso foram obtidos cDNAs de trans-sialidases de CL-Brener nas formas epimastigota e tripomastigota com o iniciador 5’UTRTCNA, os quais apresentaram UTR com tamanhos variados entre 65 – 187 pb, além da obtenção de cDNAs para esta família de proteínas, em epimastigotas CL-Brener, com o iniciador 5’TcTS, apresentando UTR variando de 171 a 221 pb, com similaridade de sequências entre elas. Com o intuito de avaliar a correspondência entre a transcrição dos RNAs de trans-sialidases e a sua tradução, houve a necessidade de identificação das proteínas. Desenvolvemos uma metodologia de lise celular e produção de extrato proteico (denominado TcS12) a partir de células de T. cruzi no estágio epimastigota. As cepas selecionadas para esse estudo foram CL-Brener (TcVI), Dm28c (TcI), Y (TcII) e 4167 (TcIV). O processo de lise celular foi otimizado para 107 parasitos/mL ressuspensos em 200 miL de tampão de lise hipotônica (por 30 minutos a 40C), associado ao sonicador de banho (por 30 minutos a 40C)...

Gene regulation in trypanosomatids is polycistronic and occurs in a post-transcriptionalway. [...] To verify the impact of UTRs presenting differentsizes in the transcription machinery and protein translation, genes from trans-sialidasefamily were selected due to its importance in the cell invasion process. Themethodological strategies involved the extraction of total RNA from T. cruzi CL-Brenerstrain, followed by cDNA synthesis, PCR, cloning and sequencing of amplified productsby Sanger and by using the 454 Junior (Next Generation Sequencing – NGS).Considering that trans-sialidase is a multi-copy gene family, this high-throughputsequencing strategy was employed in an attempt to cover the largest number of transsialidasegenes. Trans-sialidase cDNAs from CL-Brener epimastigote andtripomastigote were obtained with 5`UTRTCNA primer showing UTR sizes between 65 -187 bp. The cDNA from this protein family were also obtained with the 5’TcTS primerfrom CL-Brener epimastigotes, generating UTRs with 171 - 221 bp. Both 5’UTRpresented sequence similarities between them. In order to evaluate the correspondencebetween trans-sialidase gene transcription and translation, it was necessary toaccomplish the identification of proteins. Therefore, we developed a methodology forcell disruption, which resulted in a protein extract (referred as TcS12) from epimastigoteT. cruzi cells. The strains selected for this study were CL-Brener (TcVI), Dm28c (TcI), Y(TcII) and 4167 (TcIV). The process for lysing the cells was optimized to 107parasites/mL resuspended in 200 miL hypotonic lysis buffer (30 minutes at 4oC), followedby water bath sonication (30 minutes at 4oC). The process efficacy was confirmed byFACS, showing that near 72 percent of the cells were successfully stained with propidiumiodide solution (PI)...

Protein turnover, amino acid requirements and recommendations for athletes and active populations

Skeletal muscle is the major deposit of protein molecules. As for any cell or tissue, total muscle protein reflects a dynamic turnover between net protein synthesis and degradation. Noninvasive and invasive techniques have been applied to determine amino acid catabolism and muscle protein building at rest, during exercise and during the recovery period after a single experiment or training sessions. Stable isotopic tracers (13C-lysine, 15N-glycine, ²H5-phenylalanine) and arteriovenous differences have been used in studies of skeletal muscle and collagen tissues under resting and exercise conditions. There are different fractional synthesis rates in skeletal muscle and tendon tissues, but there is no major difference between collagen and myofibrillar protein synthesis. Strenuous exercise provokes increased proteolysis and decreased protein synthesis, the opposite occurring during the recovery period. Individuals who exercise respond differently when resistance and endurance types of contractions are compared. Endurance exercise induces a greater oxidative capacity (enzymes) compared to resistance exercise, which induces fiber hypertrophy (myofibrils). Nitrogen balance (difference between protein intake and protein degradation) for athletes is usually balanced when the intake of protein reaches 1.2 g·kg-1·day-1 compared to 0.8 g·kg-1·day-1 in resting individuals. Muscular activities promote a cascade of signals leading to the stimulation of eukaryotic initiation of myofibrillar protein synthesis. As suggested in several publications, a bolus of 15-20 g protein (from skimmed milk or whey proteins) and carbohydrate (± 30 g maltodextrine) drinks is needed immediately after stopping exercise to stimulate muscle protein and tendon collagen turnover within 1 h.

Effects of a novel calcium aluminate cement on the early events of the progressionof osteogenic cell cultures

The present study evaluated the progression of osteogenic cell cultures exposed to a novel calcium aluminate cement (CAC+) in comparison with the gold standard mineral trioxide aggregate (MTA). Cells were enzimatically isolated from newborn rat calvarial bone, plated on glass coverslips containing either CAC+ or a control MTA samples in the center, and grown under standard osteogenic conditions. Over the 10-day culture period, roundening of sample edges was clearly noticed only for MTA group. Although both cements supported osteogenic cell adhesion, spreading, and proliferation, CAC+-exposed cultures showed significantly higher values in terms of total cell number at days 3 and 7, and total protein content and alkaline phosphatase activity at day 10. The present in vitro results indicate that the exposure to CAC+ supports a higher differentiation of osteogenic cells compared with the ones exposed to MTA. Further experimental studies should consider CAC+ as a potential alternative to MTA when the repair of mineralized tissues is one of the desired outcomes in endodontic therapy.

O objetivo do presente estudo foi avaliar a progressão de cultura de células osteogênicas expostas a um novo cimento de aluminato de cálcio (CAC+) em comparação ao agregado de trióxido mineral (MTA). As células foram obtidas por digestão enzimática de calvária de ratos recém-nascidos, plaqueadas sobre lamínulas de vidro contendo em sua área central discos de CAC+ ou MTA e crescidas em condições osteogênicas por até 10 dias. Durante a cultura primária, observou-se o arredondamento das bordas das amostras de cimento apenas para MTA. Embora ambos os cimentos tenham permitido a adesão, o espraiamento e a proliferação celulares, as culturas crescidas em contato com CAC+ exibiram valores maiores de número total de células em 3 e 7 dias, e de conteúdo de proteína total e atividade de fosfatase alcalina em 10 dias. Os resultados indicam que a exposição ao CAC+ permite o desenvolvimento de uma proporção maior de células em estágios mais avançados da diferenciação osteoblástica, quando comparado ao MTA. Deve-se considerar em futuros estudos experimentais a utilização do CAC+ como um material alternativo ao MTA especialmente quando um dos objetivos do tratamento endodôntico é o de reparação dos tecidos mineralizados da região periapical.

Regulation of protein synthesis and the role of eIF3 in cancer

Maintenance of cell homeostasis and regulation of cell proliferation depend importantly on regulating the process of protein synthesis. Many disease states arise when disregulation of protein synthesis occurs. This review focuses on mechanisms of translational control and how disregulation results in cell malignancy. Most translational controls occur during the initiation phase of protein synthesis, with the initiation factors being the major target of regulation through their phosphorylation. In particular, the recruitment of mRNAs through the m7G-cap structure and the binding of the initiator methionyl-tRNAi are frequent targets. However, translation, especially of specific mRNAs, may also be regulated by sequestration into processing bodies or stress granules, by trans-acting proteins or by microRNAs. When the process of protein synthesis is hyper-activated, weak mRNAs are translated relatively more efficiently, leading to an imbalance of cellular proteins that promotes cell proliferation and malignant transformation. This occurs, for example, when the cap-binding protein, eIF4E, is overexpressed, or when the methionyl-tRNAi-binding factor, eIF2, is too active. In addition, enhanced activity of eIF3 contributes to oncogenesis. The importance of the translation initiation factors as regulators of protein synthesis and cell proliferation makes them potential therapeutic targets for the treatment of cancer.

Protein synthesis regulation by leucine / Regulação da síntese de proteína pela leucina

In vivo and in vitro studies have demonstrated that high protein diets affect both protein synthesis and regulation of several cellular processes. The role of amino acids as substrate for protein synthesis has been established in the literature. However, the mechanism by which these amino acids modulate transcription and regulate the mRNA translation via mTOR-dependent signaling pathway has yet to be fully determined. It has been verified that mTOR is a protein responsible for activating a cascade of biochemical intracellular events which result in the activation of the protein translation process. Of the aminoacids, leucine is the most effective in stimulating protein synthesis and reducing proteolysis. Therefore, it promotes a positive nitrogen balance, possibly by favoring the activation of this protein. This amino acid also directly and indirectly stimulates the synthesis and secretion of insulin, enhancing its anabolic cellular effects. Therefore, this review aimed to identify the role of leucine in protein synthesis modulation and to discuss the metabolic aspects related to this aminoacid.

Estudos in vivo e in vitro verificaram que dietas hiperprotéicas influenciam a síntese protéica e regulam vários processos celulares. O papel dos aminoácidos como substrato para a síntese de proteínas já está bem evidenciado na literatura, porém as formas como esses aminoácidos modulam a etapa da transcrição e regulam a tradução do RNAm, pela via de sinalização dependente da mTOR, ainda não estão totalmente esclarecidas. Tem-se verificado que a mTOR é uma proteína responsável por ativar uma cascata de eventos bioquímicos intracelulares que culminam na ativação do processo de tradução protéica. Dentre todos os aminoácidos, a leucina é a mais eficaz em estimular a síntese protéica, reduzir a proteólise e, portanto, favorecer o balanço nitrogenado positivo, possivelmente por favorecer a ativação desta proteína. Além disso, este aminoácido estimula direta e indiretamente a síntese e a secreção de insulina, e, assim, aumenta as propriedades anabólicas celulares. Nesse sentido, a presente revisão tem como objetivo identificar o papel da leucina na modulação da síntese protéica e abordar aspectos metabólicos relacionados a este aminoácido.

Chitinase production by Streptomyces sp. ANU 6277

Chitinase production by a terrestrial Streptomyces sp. ANU 6277 was studied under sub-merged fermentation. Chitinase production started after 24 h of incubation and reached maximum levels after 60 h of cultivation. A high level of chitinase activity was observed in the culture medium with pH 6 at 35ºC. Culture medium amended with 1 percent chitin was found to be suitable for maximum production of chitinase. An optimum concentration of colloidal chitin for chitinase production was determined. Studies on the influence of additional carbon and nitrogen sources on chitinase production revealed that starch and yeast extract served as good carbon and nitrogen sources to enhance chitinase yield.Chitinase was purified from crude enzyme extract by single step gel filtration by Sephadex G-100. Purified chitinase of the strain exhibited a distinct protein band near 45 kDa by means of SDS-PAGE.