bluuebunny/crossref_metadata_2025 huggingface dataset:
Respostas à política nacional de mobilidade urbana: comparativo entre capitais dos incentivos ao transporte público e à bicicleta
Authors: Lourenço Giovana Hardt , Dal Bosco Jr Alceu , Bernardinis Márcia De Andrade Pereira
Published: August 2019

A Política Nacional de Mobilidade Urbana (NPUM) surgiu com o objetivo de guiar as cidades em decisões que garantam mais acessibilidade e sustentabilidade com prioridades para transporte coletivo (ônibus veículos sobre trilhos) e não motorizado. O presente trabalho propõe uma sistemática de avaliação da efetividade prática da PNMU no que tange o transporte público e uso de bicicleta, através de alguns indicadores do Índice de Mobilidade Urbana Sustentável (IMUS), criado por Rodrigues da Silva et al. (2008), com modificação do método original. Foram comparadas quatro capitais brasileiras: Curitiba, São Paulo, Rio de Janeiro e Belo Horizonte. Os indicadores foram analisados temporalmente, de 2010 até 2015, para traçar uma evolução das cidades frente às premissas da Lei de Mobilidade de 2012 e compará-las utilizando o teste estatístico T-Student. Observou-se que as evoluções não têm distinção significativa, porém as médias de pontuação sim. Os resultados são inconclusivos para a comparação, devido à dificuldade de se encontrar dados abertos necessários para melhor avaliar os indicadores. Ainda assim, o IMUS se apresenta como um critério promissor de avaliação da NPUM.
Potencial de transferência de um índice de mobilidade sustentável para campus universitário
Authors: Marvulle Tan Francine , Rodrigues da Silva Antônio Nélson
Published: August 2019

O objetivo do presente trabalho é desenvolver um método para avaliação do potencial de transferência de um índice de mobilidade sustentável para campus universitário. O estudo teve início com a aplicação do IMSCamp, índice concebido por Oliveira (2015), a um novo contexto. Em seguida foi analisado o potencial de transferência do referido índice, a partir de sua concepção original, através de critérios desenvolvidos especificamente para este fim. A aplicação ocorreu no campus da USP de Piracicaba e resultou no valor de 0,491, o que não reflete boas condições de mobilidade, visto que este valor pode variar de zero (pior caso) a um (melhor caso). A avaliação seguinte também resultou em um valor que indica um baixo potencial de transferência (cerca de 40% do máximo possível) para o índice. Ou seja, a transferência do índice a um novo contexto exigiu que o mesmo fosse cuidadosamente revisto e adaptado para contemplar as novas necessidades.
Análise de deformações específicas de tração na fibra inferior de revestimento asfáltico através de instrumentação e métodos numéricos
Authors: Carvalho Nielson Tôrres Neves de , Albuquerque Fernando Silva
Published: August 2019

O uso de estimativas numéricas das tensões, deformações e deflexões resultantes do tráfego de veículos, para dimensionamento de pavimentos rodoviários, é bastante comum pela praticidade, porém pode trazer imprecisões, provocando superdimensionamento ou subdimensionamento das camadas do pavimento. Por sua vez, o uso da instrumentação, realizando medições diretas no pavimento, tende a fornecer maior exatidão. Dentre as respostas mecânicas críticas, destaca-se a deformação específica de tração na fibra inferior das camadas asfálticas, ligada ao processo de fadiga. Neste trabalho foi realizada avaliação dessa deformação medida através de sensor H-Gage, comparando em seguida com as estimativas numéricas através de modelagem com os métodos das múltiplas camadas elásticas e dos elementos finitos. Ambos os modelos numéricos apresentaram estimativas bem discrepantes das medições de campo, tendo diferença ainda maior nas respectivas previsões de vida de fadiga. O método das múltiplas camadas elásticas apresentou tendência a estimar menor vida de fadiga, enquanto o modelo proposto em elementos finitos tendeu a estimar maior vida de fadiga, à medida que se reduzia o atrito entre camadas.
Avaliação do uso de cinzas de carvão mineral produzidas em usina termelétrica na construção de pavimentos
Authors: Vasconcelos Sarah Denise , Barroso Suelly Helena De Araújo , Vieira Francisco De Assis Franco , Almeida Helmer Boris Fernandes
Published: August 2019

O aproveitamento de rejeitos produzidos em centrais termelétricas pode representar uma excelente alternativa para construção de pavimentos sustentáveis. O estudo proposto tem por objetivo avaliar a utilização de cinzas de carvão provenientes de uma termelétrica, localizada na Região Metropolitana de Fortaleza, em camadas dos pavimentos. Inicialmente foram caracterizadas as cinzas que estavam dispostas em um módulo da termelétrica em estudo e foi comprovado que os materiais, além de heterogêneos, não tinham comportamento mecânico satisfatório. Sendo assim, foram produzidas em laboratório misturas de cinza-solo-cal submetidas aos diferentes ensaios (compactação, módulo do resiliência, deformação permanente, resistência à compressão simples e resistência à tração por compressão diametral) de interesse na pavimentação. Posteriormente, foram dimensionadas pelo método mecanístico-empírico estruturas de pavimentos com as misturas que apresentaram resultados mecânicos mais satisfatórios. Concluiu-se que o reaproveitamento dos resíduos da termelétrica na pavimentação representa uma alternativa viável para a construção de pavimentos econômicos e autossustentáveis.
Efeito da aderência entre camadas na previsão de desempenho de pavimentos asfálticos
Authors: Dos Santos Anthony Gomes , De Assis Rosuel Krum Mathias , Fernandes Júnior José Leomar
Published: August 2019

Os métodos mecanísticos de projeto de pavimentos admitem perfeita aderência entre as camadas, o que pode não ocorrer no campo. Neste trabalho, com uso do programa de elementos finitos ABAQUS, são analisados os efeitos da condição de interface das camadas (aderência completa, parcial e nula) no comportamento mecânico de pavimentos. As análises foram realizadas para diferentes níveis de rigidez do material asfáltico e a estimativa do desempenho de pavimentos flexíveis, tanto a fadiga das camadas asfálticas quanto a deformação permanente, foi baseada em equações empírico-mecanísticos que consideram, respectivamente, as respostas estruturais: deformação horizontal de tração na fibra inferior das camadas asfálticas e deformação vertical de compressão no topo do subleito. Os resultados mostram que as respostas estruturais são mais sensíveis à condição de aderência na interface entre as camadas asfálticas, para a qual contribui a pintura de ligação, do que na segunda interface. A condição de interface entre as camadas afeta mais a vida do pavimento à fadiga que à deformação permanente e em ambos os casos a consideração de perfeita aderência entre as camadas resulta na previsão de melhor desempenho dos pavimentos. Portanto, o surgimento prematuro de defeitos pode estar relacionado, também, à fraca aderência entre camadas.
Intermittent demand forecasting for aircraft inventories: a study of Brazilian’s Boeing 737NG aircraft´s spare parts management
Authors: Silva Jersone Tasso Moreira , Santos Luiz Henrique , Dias Alexandre Teixeira , Tadeu Hugo Ferreira Braga
Published: August 2019

Este estudo tem como objetivo avaliar cinco métodos de previsão para demanda intermitente usando uma série histórica de consumo de peças sobressalentes da aeronave 737 Next Generation, fabricado pela Boeing, da maior frota aérea brasileira gerenciada pela VRG Airline Company S/A. Os métodos de Winter, Croston, Single Exponential Smoothing, Weight Moving Average e Método de Distribuição de Poisson foram testados em um histórico de 53 peças sobressalentes e cada uma delas possui um histórico de demanda de trinta e seis meses (janeiro de 2013 a dezembro de 2015). Os resultados mostraram que os métodos Weight Moving Average, Distribuição de Poisson e Croston apresentaram os melhores ajustes. Além disso, observou-se que a maior parte das demandas por peças sobressalentes apresentaram um padrão smooth ao contrário do resultado obtido pelo estudo de Ghobbar and Friend (2003) que apresentou um padrão lumpy. Por outro lado, tem-se que o Método de Winter apresentou-se como o de pior ajuste em ambos os estudos. Conclui-se que os métodos de Weight Moving Average e Distribuição de Poisson são os mais adequados para avaliar a demanda intermitente para o caso da VRG Airline Company S/A.
Avaliação do potencial de integração entre o uso e ocupação do solo e o transporte de carga em um recorte urbano de São Paulo
Authors: Montmorency Silva Thiago Canhos , Marins Karin Regina De Castro
Published: August 2019

O objetivo desse artigo é realizar uma avaliação integrada entre o transporte urbano de carga e o uso e ocupação do solo, por meio da aplicação do modelo Freturb em um recorte urbano na cidade de São Paulo. Os resultados indicaram haver interdependências entre os parâmetros do transporte urbano de carga (tipos de veículos comerciais, condições de carga e descarga e quantidades de movimentações) e do uso e ocupação do solo (perfil econômico, dimensões dos estabelecimentos alimentícios, número de empregos e de habitantes), revelando quais aspectos do transporte urbano da carga impactam diretamente no uso do espaço urbano e quais elementos do uso e ocupação do solo condicionam aspectos da logística e do tráfego urbano, em um contexto urbano brasileiro. Entende-se que as inter-relações demonstradas sejam fundamentais para aprimorar políticas urbanas relacionadas à mobilidade urbana e ao uso e ocupação do solo, ao fomentar o planejamento e a gestão intersetoriais e interdisciplinares em cidades brasileiras.
Método para definição de rede de rotas cicláveis em áreas urbanas de cidades de pequeno porte: um estudo de caso para a cidade de Bariri-SP
Authors: Monari Marcelo , Segantine Paulo Cesar Lima
Published: August 2019

Neste trabalho é apresentado um método para definição de rede de rotas cicláveis em áreas urbanas de cidades de pequeno porte. As rotas cicláveis são identificadas de maneira a minimizar a somatória das impedâncias dos segmentos viários que promovem a ligação entre pares origem-destino de interesse ao transporte cicloviário. Estas impedâncias são calculadas considerando-se, para cada segmento, suas respectivas avaliações globais do nível de estresse para ciclistas, e fatores relacionados à declividade e à existência ou possibilidade de inserção de infraestruturas cicloviárias. O estudo de caso, conduzido à cidade de Bariri-SP, mostrou a viabilidade de aplicação do método proposto, permitindo a identificação de 35 rotas cicláveis compostas principalmente por segmentos viários com pequenas declividades e boas avaliações globais do nível de estresse. A reincidência destas rotas permitiu a definição de uma rede composta por 19 eixos viários potencialmente cicláveis distribuídos ao longo de toda a área de estudo.
Classificação de indivíduos segundo comportamento individual relativo a viagens a partir de dados em painel obtidos por smartphones
Authors: Assirati Lucas , Pitombo Cira Souza
Published: August 2019

A caracterização comportamental relativa a viagens é uma questão importante nas análises baseadas em atividades e, comumente, é a variável dependente nos modelos de estimativa de demanda por transportes. A classificação individual, segundo comportamentos relacionados aos deslocamentos, pode ser realizada com dados seccionais, considerando diferentes fatores como distâncias, modos utilizados e atividades realizadas, ou com dados em painel, através de valores médios nos múltiplos dias ou atividades frequentes, por exemplo. Dados em painel constituem importante ferramenta em análises comportamentais relativas às viagens urbanas, propiciando dimensão analítica extra relativo à heterogeneidade temporal individual. Todavia, a obtenção desses dados não é trivial, demandando recursos monetários e de tempo. Assim, o objetivo principal deste trabalho é classificar indivíduos segundo comportamento relativo a viagens a partir de dados em painel. O objetivo secundário associa-se à obtenção do painel através de smartphones. A potencialidade da proposta é validada por um estudo de caso contemplando estudantes universitários em São Carlos – SP, Brasil. Mediante dados fornecidos pelos estudantes, utilizou-se o algoritmo k-médias considerando quatro variáveis associadas às viagens realizadas em três dias úteis consecutivos. Obtiveram-se três grupos comportamentais distintos com diferenças quanto ao grau de motorização, recorrência de localidades, número de viagens realizadas e distâncias médias percorridas.
Avaliação da irregularidade longitudinal do pavimento sobre pontes usando acelerações medidas por smartphones
Authors: Muliterno Betina Knebel , Muliterno Junior Fernando , Pravia Zacarias Martin Chamberlain
Published: August 2019

O objetivo deste trabalho foi obter o Índice de Irregularidade Internacional (IRI) de pavimentos sobre pontes utilizando smartphones, estes equipados com acelerômetros, possibilitando a aquisição de dados de aceleração e a determinação da irregularidade através de procedimentos numéricos, consistindo na dupla integral dos dados de aceleração vertical. Foram realizadas medições em três pontes, por meio de um veículo percorrendo as mesmas, equipado com dois telefones móveis diferentes com taxa de aquisição de 100 Hz. Um dos dispositivos foi disposto no interior do veículo e outro na bandeja do sistema de suspensão. Utilizou-se outra abordagem, o Método de Nível e Mira, para a calibração da medição do tipo resposta nas pontes. As conclusões mostram uma boa convergência entre os métodos apresentados para obtenção de perfis de superfície de pontes, demostrando bom indício do uso dos smartphones como método alternativo simples e econômico, revelando fácil operação e boa produtividade.
Estudo da aplicação da cinza de caldeira a coque nas camadas de base e sub-base de pavimentos
Authors: Silveira Virlene Leite , Guimarães Antônio Carlos Rodrigues , Castro Carmen Dias
Published: August 2019

Este estudo apresenta a caracterização de cinzas obtidas do processo de beneficiamento do níquel em usina siderúrgica, tendo como objetivo principal, avaliar sua aplicabilidade para estabilização de solos em camadas de base e sub-base de pavimentos rodoviários, através da mistura destas a um solo regional e cal. Foram realizados ensaios de caracterização química, física e mecânica, para o solo puro e para o mesmo solo com a adição de diferentes teores de cinzas e cal em diferentes tempos de cura a fim de se verificar a ocorrência de reações pozolânicas, tais que resultassem em aumento da rigidez do solo. A análise mecânica constituiu de ensaio de módulo de resiliência em corpos de prova contendo 10%, 15% e 20% de cinza em substituição a massa seca do solo e 8%, 13% e 18% de cinza e 2% de cal em substituição a massa seca da cinza. Como resultados, encontrou-se que a cinza de caldeira a coque provoca acréscimo da rigidez das misturas com aumento do tempo de cura, podendo ter seu efeito potencializado pela ação da cal, efeito observado através dos ensaios triaxial de cargas repetidas. Os resultados obtidos para todos os teores de cinza foram satisfatórios, ressaltando o emprego positivo do material para aplicação em camadas de pavimentos em vias de baixo a médio tráfego.
Compreensão da formação de NO2 proveniente das operações de transporte urbano e suas relações com agentes causais
Authors: Ribeiro Jefferson Pereira , Cassiano Demostenis Ramos , Bertoncini Bruno Vieira , Castelo Branco Verônica Teixeira Franco , Sousa Francelino Franco Leite de Matos , Quintanilha Wendy Fernandes Lavigne , Cavalcante Rivelino Martins , Santiago Ithala Saldanha de , Fernandes Gabrielle Melo
Published: August 2019

Os problemas oriundos da mobilidade urbana estão relacionados ao crescimento acelerado e não planejado das cidades, à distribuição espacial das atividades, ao crescimento expressivo do uso do automóvel, as deficiências do transporte público e a ocorrência de impactos negativos nos âmbitos sociais e ambientais, tendo como um dos principais impactos a concentração de NO2. Dessa forma, o presente trabalho tem por objetivo avaliar a influência dos fatores (gabarito das edificações, uso do solo e volume de tráfego) nas concentrações de NO2. Para tal, a complexidade do uso do solo urbano foi traduzida por meio do indicador que mede a diversidade do uso do solo urbano, entropia. As concentrações de NO2 foram medidas através de um método de amostragem passiva. Como principais resultados verificou-se que o volume de tráfego veicular aliado a elevada entropia se relacionam diretamente com as concentrações de NO2. O gabarito das edificações se relaciona com estas concentrações mas não de forma tão direta, pois tal fator pode ser suprimido pelo tráfego veicular ou influenciado por fenômenos naturais que facilitam ou não a dispersão de NO2.
Fatores influentes para a adesão de alunos a sistema de carona dinâmica em campus universitário
Authors: De Souza Silva Laize Andréa , De Andrade Maurício Oliveira , Maia Maria Leonor Alves
Published: August 2019

Este artigo objetiva a investigação dos fatores que influenciam a oferta de carona por alunos do Campus Recife da Universidade Federal de Pernambuco. Investigam-se os critérios relevantes para a adesão a um hipotético sistema dinâmico de caronas. São ainda sugeridas medidas mais eficazes para ampliação do uso dessa alternativa de transporte. O método de análise utilizado foi a Regressão Logística. Os resultados obtidos mostram que fatores mutáveis, inerentes ao arranjo da carona, e psicológicos estão relacionados ao modo dinâmico. Enquanto o único critério fixo que afeta a probabilidade de uso do sistema proposto é o sexo do usuário, seis critérios intrínsecos do próprio sistema apresentam-se significativos: i) perfil do usuário; ii) possibilidade de ser passageiro através do sistema; iii) questões ambientais; iv) maior interação social; v) redução da flexibilidade; e vi) compartilhar viagens com desconhecidos. Conclui-se que o uso da carona pode ser aumentado a partir de sistemas de caronas dinâmicas. Além disso, junto às plataformas de conexão, é importante que sejam adotadas políticas de caráter informativo e educacional.
Development of dual priming oligonucleotide-polymerase chain reaction (DPO-PCR) for detection of wheat component in foods
Authors: Su Yan , Li Rong
Published: August 2019

This research is aimed to establish a DPO-PCR method for rapid detection of wheat component in foods. A pair of highly specific DPO primers was designed using the wheat housekeeping gene GAG56D as the target gene. By optimizing the PCR reaction system, the DPO-PCR detection method for wheat component in food was established, and the specificity and sensitivity of the method were determined. The established DPO-PCR method was highly specific, and there was no cross-reactivity to 22 control samples. The sensitivity was high, and the minimum detection limit was 1 ng/uL. Tests on commercially available product samples showed that the method can effectively detect wheat component of foods. The DPO-PCR method established in this study is simple and accurate, and provides an effective detection method for wheat component in foods.
Gentiopicrin exerts anti-rheumatic effect in human fibroblast-like synoviocytes via inhibition of p38MAPK/NF-ÎºB pathway
Authors: Zhang Nian , Jiang Yi , Mu Fangxiang , Wu Hong , You Qingxia
Published: August 2019

To investigate the effect of gentiopicrin on the expressions of inflammatory factors in human fibroblast-like synoviocytes (HFLS) and the underlying mechanism. Human fibroblast-like synoviocytes (HFLS) were cultured in vitro at 37 °C in Dulbecco's modified Eagle's medium (DMEM) supplemented with 5 % fetal bovine serum (FBS) in a humidified incubator containing 5 % CO2. Cell viability was determined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-tetrazolium bromide (MTT) assay, while real-time quantitative polymerase chain reaction (qRT-PCR) was used to determine the expressions of interleukin 1Î² (IL-1Î²) and interleukin 6 (IL-6) mRNAs. The expressions of p38 mitogen-activated protein kinase (p38 MAPK) and nuclear factor kappa light chain enhancer of activated B cells (NF-ÎºB) were determined using Western blotting. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of IL-1Î² and IL-6 in cell lysate. Treatment with 5-25 Î¼M gentiopicrin did not significantly affect the number of viable cells, when compared with control group (p > 0.05). However, at 50 and 100 Î¼M gentiopicrin, the number of viable cells were significantly increased, relative to control group (p < 0.05). Results of qRT-PCR showed that the expression levels of IL-1Î² and IL-6 mRNAs were significantly higher in TNF-Î± group than in control group (p < 0.05). However, treatment with gentiopicrin significantly and dose-dependently decreased their expression levels compared with TNF-Î± group (p < 0.05). Western blotting results showed that the expressions of p-p38MAPK and NF-ÎºB-p65 proteins were significantly upregulated in TNF-Î± group, when compared with control group (p < 0.05). However, treatment with gentiopicrin significantly and dose-dependently down-regulated the expression of these proteins compared with TNF-Î± group (p < 0.05). The levels of IL-1Î² and IL-6 in cell lysate were significantly higher in TNF-Î± group than in control group (p < 0.05). However, treatment with gentiopicrin, and p38MAPK/NF-ÎºB pathway inhibitors (SB203580 and BAY11-7082) significantly reduced the levels of these inflammatory factors compared with TNF-Î± group (p < 0.05).  Gentiopicrin has therapeutic potential for Rheumatoid arthritis (RA  ) through a mechanism involving the inhibition of p38MAPK/NF-ÎºB pathway.
Sesamolin exerts anti-proliferative and apoptotic effect on human colorectal cancer cells via inhibition of JAK2/STAT3 signaling pathway
Authors: Wu Di , Wang Xin-Ping , Zhang Wei
Published: August 2019

Colorectal cancer (CRC) is a common malignant tumor that seriously threatens human health and quality of life. At present, the search for safe and more effective treatment for CRC has become necessary. The present study investigated the anti-proliferative and apoptotic effects of sesamolin on human colorectal cancer (HCT116) cells, and the underlying mechanism. Cell proliferation was determined using MTT assay, while the expressions of JAK2, STAT3 and p-STA3 were determined using Western blotting. The levels of expression of matrix metalloproteinases-1, 2 and 9 (MMP1, MMP2 and MMP9) were determined using real-time quantitative polymerase chain reaction (qRT-PCR). The degree of migration and invasion of the cells was assessed using wound healing assay. The results of MTT assay showed that sesamolin significantly and time- and dose-dependently inhibited the proliferation of HCT116 cells (p < 0.05). Treatment of HCT116 cells with sesamolin significantly inhibited their migratory ability (p < 0.05). The expressions of p-JAK2 and p-STAT3 were significantly down-regulated 48 h after 20 µM of JAK2 specific inhibitor (AG490) was added to HCT116 cells (p < 0.05). The expression of p-STAT3 was also significantly and dose-dependently down-regulated 6 h after treatment of HCT116 cells with sesamolin (p < 0.05). Sesamolin and AG490 had synergistic effect and their combination significantly down-regulated the expression of p-STAT3, when compared with sesamolin alone (p < 0.05). Treatment of HCT116 cells with sesamolin significantly and dose-dependently reduced the levels of IL-6-induced expressions of MMP-1, MMP-2 and MMP-9 (p < 0.05). These results suggest that sesamolin induces apoptosis in HCT116 cells and prevents cell invasion via inhibition of the JAK2/STAT3 signaling pathway.
Capsular Polysaccharide Biosynthesis from Recombinant E. coli and Chondroitin Sülfate Production
Authors: Erenler Ayse Sebnem
Published: August 2019

Chondroitin sulfate (CS) is an important biomedical product. CS is the basic structural component of the mammalian extracellular matrix and is widely used in many applications in the fields of medicine, veterinary medicine, pharmaceuticals and cosmetics. For CS production, mainly animal sources are used. However, in today's conditions, due to various risks and artificial synthesis, there has been an increase in alternative sources of production methods for CS, instead of using animal resources. In this study as a powerful alternative microbial production of CS has been targeted. By using recombinant E. coli strains to integrate VHb /vgb+ and kfo+ systems, the aim was to obtain high purity CS from reliable biotechnological processes. Plasmid pUC8:15 bearing the vgb gene region, and plasmid pETM6-PACF carrying the kfoA, kfoC and kfoF genes responsible for chondroitin synthesis, were transferred to E. coli bacteria. Microbial CS was obtained by adding sulfate groups to chondroitin acquired after the treatments. The results were confirmed by HPLC and NMR analyses. The product, compared to its counterparts, was found to be an effective drug, potentially with a low molecular weight  value.
Prediction of Strength of Remixed Concrete by Application of Orthogonal Decomposition, Neural Analysis and Regression Analysis
Authors: Bidkar K.L. , Jadhao P.D.
Published: August 2019

Compressive strength is the foremost property of concrete which is influenced by a number of parameters. These parameters plays important role for the characteristics achieved by concrete. Orthogonal decomposition, neural analysis and regression analysis tools can be utilized where the dependence and independence of these parameters to be considered. In this paper these analyses are considered for remix concrete, in which apart from the cement contents, w/c ratio, proportions of C.A., F.A., the other parameters like blend ratio (r=Q
o
/Q
f
, Q
o
=quantity of old partially set concrete, Q
f
=quantity of fresh concrete) time lag ( time between preparation and placing of concrete) also plays the important role.
Direct coupling analysis improves the identification of beneficial amino acid mutations for the functional thermostabilization of a delicate decarboxylase
Authors: Peng Martin , Maier Manfred , Esch Jan , Schug Alexander , Rabe Kersten S.
Published: August 2019

The optimization of enzyme properties for specific reaction conditions enables their tailored use in biotechnology. Predictions using established computer-based methods, however, remain challenging, especially regarding physical parameters such as thermostability without concurrent loss of activity. Employing established computational methods such as energy calculations using FoldX can lead to the identification of beneficial single amino acid substitutions for the thermostabilization of enzymes. However, these methods require a three-dimensional (3D)-structure of the enzyme. In contrast, coevolutionary analysis is a computational method, which is solely based on sequence data. To enable a comparison, we employed coevolutionary analysis together with structure-based approaches to identify mutations, which stabilize an enzyme while retaining its activity. As an example, we used the delicate dimeric, thiamine pyrophosphate dependent enzyme ketoisovalerate decarboxylase (Kivd) and experimentally determined its stability represented by a T50 value indicating the temperature where 50% of enzymatic activity remained after incubation for 10 min. Coevolutionary analysis suggested 12 beneficial mutations, which were not identified by previously established methods, out of which four mutations led to a functional Kivd with an increased T50 value of up to 3.9°C.
Clinical, biochemical and gender characteristics of 97 prepubertal children with premature adrenarche
Authors: Santos-Silva Rita , Costa Carla , Castro-Correia Cíntia , Fontoura Manuel
Published: August 2019

Background

Premature adrenarche (PA) is defined as the appearance of clinical signs of androgen action associated with levels of dehydroepiandrosterone sulfate (DHEAS) ≥40 μg/dL, before age 8 years in girls and 9 years in boys, without breast or testicular enlargement. The aim of this study was to characterize a population of prepubertal Caucasian children with PA and to compare them with regard to gender and body mass index (BMI) (normal BMI vs. overweight/obesity).

Methods

We performed a cross-sectional study of Portuguese Caucasian prepubertal children followed, due to PA, in pediatric endocrinology clinics of a university hospital.

Results

Eighty-two girls and 15 boys were included (mean age at evaluation: 7.4 ± 1.3 years). The mean birth weight was 2990 ± 689 g; only two children were small for gestational age. Girls presented premature pubarche at a younger age (median [interquartile range (IQR)] 6 (5–6) years vs. 7 (7–8) years in boys; p < 0.001). No gender differences were found for gestational age, birth weight, maternal age at menarche, anthropometry, bone age advancement or androgen levels. The majority of the subjects were overweight or obese (59%). Overweight/obese PA children were taller and had a more advanced bone age than normal-BMI PA children. Overweight/obese children presented higher levels of DHEAS and androstenedione. Bone age advancement and DHEAS were correlated (r = 0.449; p = 0.05).

Conclusions

We found no evidence of reduced fetal growth. Girls presented premature pubarche at a younger age. No major gender differences in androgen levels were found in prepuberty. Obese and overweight PA children tend to be taller, have a more advanced bone age and higher levels of androgens than normal-BMI PA children.
Micro-RNA 196a2 expression and miR-196a2 (rs11614913) polymorphism in T1DM: a pilot study
Authors: Ibrahim Alshaymaa A. , Ramadan Abeer , Wahby Aliaa Ahmed , Hassan Mirhane , Soliman Hend M. , Abdel Hamid Tamer A.
Published: August 2019

Background

Recent emerging evidence supports the role of miR-196a2 in various human diseases. However, its role in type 1 diabetes mellitus (T1DM) is still underestimated. We aimed, for the first time, to investigate the expression of miR-196a2 in T1DM and the association of miR-196a2 (rs11614913) polymorphism with susceptibility of T1DM in a sample of patients from Cairo, Egypt.

Methods

The study included 150 patients and 150 healthy subjects. Evaluation of rs11614913 genotypes and miR-196a2 expression was done using the allelic discrimination and quantitative reverse transcriptase polymerase chain reaction (PCR) method, respectively.

Results

The Hardy-Weinberg equilibrium of single nucleotide polymorphism(SNP) was detected among controls (p = 0.2). Our results revealed that the TT genotype was more frequent in patients (22.6%) than controls (10%) while the CC genotype was more frequent in controls (47.3%) than patients (39.3%) (p = 0.01). The frequency of the T allele was significantly higher in patients than in controls (41.7 vs. 31.3%), while the C allele was more frequent in controls (p = 0.008). After adjustment for traditional risk factors, the association of the TT genotype with T1DM remained significant (TT vs. CC, odds ration [OR] = 3.2, 95% confidence interval [CI]: 1.4–7.4, p = 0.005). Power analysis of the data yielded a statistical power of 80% for the miR-196a2 rs11614913 with T1DM. Relative expression of miR-196a2 showed significant decrease in patients compared to controls (median = 0.09, 0.5, interquartile range [IQR] = 0.03–1.6, 0.1–2.1). However, miR-196a2 expression showed no significant difference between different rs11614913 genotypes (p = 0.5).

Conclusions

Our findings demonstrated that miR-196a rs11614913 is associated with T1DM and decreased expression of miR-196a2 may play a role in pathogenesis of T1DM.
Phenylketonuric patients represent their disease
Authors: Iakovou Kostas , Schulpis Kleopatra
Published: August 2019

Background

Phenylketonuria (PKU) is a metabolic disorder resulting in high phenylalanine (Phe) blood concentrations due to the deficiency of the hepatic enzyme Phe hydroxylase. High Phe levels are currently treated with diet, thus avoiding mental retardation, psychomotor delay, etc. The aim of this study was to find out the self-presentation of the disease by the patients.

Methods

A total number of 110 patients self-represented their disease: food allergy (Phe allergy) n = 34/110 (30.9%), health problems n = 31/110 (28.2%), severe health problems n = 26/110 (23.6%), very severe health problems n = 19/110 (17.3%). The Wechsler Intelligence Scale for Children (WISC III) psychometric test was utilized for the evaluation of IQ scores of each participant. A special questionnaire was used focusing on quality of life (QL), social discrimination (SD) and anxiety or depression (AD).

Results

IQ scores were found to be higher in food allergy self-represented patients than those of the other tested groups. The lowest IQ scores were observed in those who represented very severe disease. QL, SD and AD were not observed in the food allergy groups. On the contrary, those who self-presented as very severe diseased patients felt high a disturbance of QL, SD and AD.

Conclusions

The food allergy group seemed to be a very clever excuse for adhering to PKU treatment resulting in high IQ scores and absence of the mentioned upsets. In contrast, in the other groups of patients’ IQ scores were found to be lower than that of food allergy group. Damage of QL, SD, AD were also apparent.
Severe craniofacial trauma after multiple pistol shots
Authors: Haller Ingrid , Lederer Wolfgang , Glodny Bernhard , Wiedermann Franz J.
Published: August 2019

A 48-year-old woman suffered from life-threatening injuries in head and chest caused by six pistol shots fired at close range in an attempted homicide. We report here on our successful airway management and bleeding control at the scene of crime and the multidisciplinary surgical treatment of the associated head and neurovascular injuries.
Drug Resistance and the Prevention Strategies in Food Borne Bacteria: An Update Review
Authors: Hashempour-Baltork Fataneh , Hosseini Hedayat , Shojaee-Aliabadi Saeedeh , Torbati Mohammadali , Alizadeh Adel Mirza , Alizadeh Matin
Published: August 2019

Antibiotic therapy is among the most important treatments against infectious diseases and has tremendously improved effects on public health. Nowadays, development in using this treatment has led us to the emergence and enhancement of drug-resistant pathogens which can result in some problems including treatment failure, increased mortality as well as treatment costs, reduced infection control efficiency, and spread of resistant pathogens from hospital to community. Therefore, many researches have tried to find new alternative approaches to control and prevent this problem. This study, has been revealed some possible and effective approaches such as using farming practice, natural antibiotics, nano-antibiotics, lactic acid bacteria, bacteriocin, cyclopeptid, bacteriophage, synthetic biology and predatory bacteria as alternatives for traditional antibiotics to prevent or reduce the emergence of drug resistant bacteria.
Anti-aging and Sunscreens: Paradigm Shift in Cosmetics
Authors: Shanbhag Shreya , Nayak Akshatha , Narayan Reema , Nayak Usha Yogendra
Published: August 2019

Skin, being one of the vital organs and a protective barrier needs to be pampered and taken care of from early childhood. It is the most visible and the widest exposed organ and by far reflects the general health condition and the aging process in humans. Both intrinsic and extrinsic factors contribute to this complex biological process of skin aging. In recent times, skin health and its beauty is perceived as an indicator of one’s health which has resulted in an increasing demand for anti-aging products. Exposure to UV radiation is considered to be one of the factors responsible for aging termed as photoaging. In this review, we have discussed the various factors which may accelerate the process of skin aging. Various approaches and strategies to delay the process of skin aging have been emphasized upon. The patents filed in the area of anti-aging and sunscreen products have also been reviewed to gain an insight into the new formulations which have been developed as an anti-aging product. There has been a tremendous rise in the cosmetic and cosmeceuticals market with products having a dual activity of anti-aging and sun protection. Research is constantly on the rise to ensure the safety of these products. Alternatives to the current topical application of sunscreen are being considered to overcome the drawback of reapplication of the sunscreen often which can be a boon to the cosmeceutical market.
5-HIAA as a Potential Biological Marker for Neurological and Psychiatric Disorders
Authors: Jayamohananan Hridya , Manoj Kumar Maneesha Kalappurackal , T P Aneesh
Published: August 2019

Neurological and psychiatric disorders occur in about 6 percent of the global population indicating a significant amount of people suffering from neurological disorder on a varying range in day to day life. On an extensive view, there is a critical requirement for the development of an alternative biomarker for these conditions. The thwart found in developing a biomarker is the difficulty in identifying a serum biomarker as these are mostly limited to the central nervous system (CNS). Serotonin being a neurotransmitter synthesized in the raphe nuclei of the brain could serve as an alternative biomarker. Here, the limitation is that it’s quickly metabolized by the mitochondrial enzyme MAO to 5-hydroxy indole acetic acid (5HIAA). This subsequent metabolite can be used for the analysis of serotonin levels in brain by analysing its concentration in the cerebrospinal fluid (CSF). Many theories suggest that the variations in serotonin level could lead to the development of many neurological and psychiatric disorders like Alzheimer’s disease (AD), schizophrenia, depression and so on. A decreased level is noticed in these patients but this could either be due to decreased production or increased reuptake of serotonin from the neuronal synapses. For instance, we know that a patient with depression shows a significant reduction in the levels of 5HIAA, due to the location of the raphe nuclei within regions of memory and cognition. Similarly, it does shows variation in AD and mild cognitive disorder. Evolving of 5HIAA as a biomarker, could be more delicate and enhanced strategy for monitoring these disorders.
Assessment of the Effect of PLGA Co-polymers and PEG on the Formation and Characteristics of PLGA-PEG-PLGA Co-block Polymer Using Statistical Approach
Authors: Sulaiman Teuku Nanda Saifullah , Larasati Dwi , Nugroho Akhmad Kharis , Choiri Syaiful
Published: August 2019

Purpose: To assess the effect of the lactic acid (LA)-to-glycolic acid (GA) molar ratio and polyethylene glycol (PEG) concentration on the formation of poly-lactide co-glycolide acid (PLGA)-PEG-PLGA co-block polymers simultaneously using statistical approach. Methods: A 22 full factorial design with the addition of a point in the center of the design, namely curvature, was applied. Fourier transform infrared (FTIR), differential scanning calorimetry (DSC), and nuclear magnetic resonance (NMR) were performed to confirm the formation of the co-block polymer. Simvastatin (SMV), a drug model was incorporated into the nano-polymeric micellar (NpM) of PLGA-PEG-PLGA followed by solubility phase, particle size, zeta potential, and entrapment efficiency characterizations. Results: FTIR, DSC, and NMR successfully confirmed the formation of co-block polymers. Solubility of SMV increased from 2 to 44-folds depending on co-block concentration with entrapment efficiency of 59%-80%. The NpM had size in the range of 206 to 402 nm with negative zeta potential. LA to GA ratio had greater effect on particle size reduction and increasing of co-polymer length. In addition, it had higher contributions on increasing of solubility and entrapment efficiency of SMV than PEG. Conclusion: According to these findings, the LA to GA ratio and PEG concentration gained a great consideration in order to prepare the PLGA-PEG-PLGA co-block which fulfilled the quality target product profile of NpM delivery system.
Nanoemulsions of Green Tea Catechins and Other Natural Compounds for the Treatment of Urinary Tract Infection: Antibacterial Analysis
Authors: Kaur Atinderpal , Gabrani Reema , Dang Shweta
Published: August 2019

Purpose: Nanoemulsions (NEs) of polyphenon 60 (P60) and cranberry (NE I) and P60 and curcumin (NE II) were prepared with the aim to enhance anti-bacterial potential and to understand the mechanism of anti-bacterial action of the encapsulated compounds. Methods: To evaluate the antibacterial potential of the developed NE, microtiter biofilm formation assay was performed. The cytotoxicity analysis was done to assess the toxicity profile of the NEs. Further antibacterial analysis against uropathogenic strains was performed to check the developed NEs were effective against these strains. Results: In microtiter dish biofilm formation assay, both NE formulations inhibited the growth more effectively (Av. % inhibition ~84%) as compared to corresponding aqueous solution (Av. % inhibition ~64%) and placebo (Av. % inhibition ~59%) at their respective minimum inhibitory concentration (MIC) values. Cytotoxicity analysis using 3-(4,5-Dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT assay) showed that the formulations were nontoxic to Vero cells. The antibacterial studies against uropathogenic resistant strains also showed that NEs effectively inhibited the growth of bacterial strains. Conclusion: From different studies it was concluded that both the NE’s were able to inhibit bacterial strains and could be further used for the treatment of urinary tract infection (UTI). The antibacterial activity of developed NEs showed that these could be used as alternative therapies for the treatment of UTI.
Determination of Methyldopa and Paracetamol in Pharmaceutical Samples by a Low Cost Genipa americana L. Polyphenol Oxidase Based Biosensor
Authors: Antunes Rafael Souza , Thomaz Douglas Vieira , Garcia Luane Ferreira , Gil Eric de Souza , Sommerset Vernon Sydwill , Lopes Flavio Marques
Published: August 2019

Purpose: Jenipapo fruit (Genipa americana L) is a natural source of polyphenol oxidases (PPOs) whose potential in pharmaceutical analysis is noteworthy. Henceforth, this work reports the electrochemical study of a low-cost PPO-based biosensor produced from the crude extract of Jenipapo fruits and accounts a practical approach to employ this biosensor in the determination of methyldopa and paracetamol in pharmaceutical samples. Methods: In order to investigate the electrochemical properties of the biosensor, theoretical and practical approaches were employed, and both samples and the biosensor were analyzed through electrochemical impedance spectroscopy (EIS) and voltammetric techniques, namely: differential pulse voltammetry (DPV) and cyclic voltammetry (CV). Results: showcased that the biosensor presented good analytical features, as well as low detection limits (8 μmol L-1 for methyldopa and 5 μmol L-1 for paracetamol). The relative standard deviation was less than 5% mid-assay. Conclusion: The use of this biosensor is a reliable, low cost and useful alternative in the pharmaceutic determination of phenolic drugs (e.g. methyldopa and paracetamol).
Two Simple Methods for Optimizing the Production of "Difficult-to-Express" GnRH-DFF40 Chimeric Protein
Authors: Barazesh Mahdi , Mostafavipour Zohreh , Kavousipour Soudabeh , Mohammadi Shiva , Mokarram Pooneh
Published: August 2019

Purpose: GnRH-DFF40 (gonadotropin releasing hormone - DNA fragmentation factor 40) is a humanized recombinant immunotoxin and serves as a prospective candidate for targeted therapy of gonadotropin releasing hormone receptor (GnRHR) overexpressing malignancies. However, its production in Escherichia coli in a soluble and functional form still remains a challenge. Here we introduce two successful and reproducible conditions for production and purification of "difficult-to-express" GnRH-DFF40 protein. Methods: A synthetic codon optimized GnRH-DFF40 fusion gene was cloned in pET28a plasmid. Two methods including high cell density IPTG induction (HCDI) and autoinduction method (AIM) with a focus on obtaining high cell density have been investigated to enhance the protein production in (E. coli). Moreover, to obtain higher protein production several factors in the AIM method including carbon sources, incubation time and temperature, plasmid stability and double colony selection, were optimized. Results: Remarkable amounts of soluble GnRH-DFF40 protein were achieved by both methods. Cell density and protein yields in AIM was about 1.5 fold higher than that what obtained using HCDI. Initial screening showed that 25ºC is better to achieve higher protein production in both methods. pH alterations in AIM were maintained in a more constant level at 25ºC and 37ºC temperatures without any detrimental effects on cell growth during protein production phase up to 21 hours after incubation. Plasmid stability during growth and expression induction phase was maintained at a high level of 98% and 96% for AIM and HCDI methods, respectively. After parameter optimization and double colony selection in AIM, a very high yield of recombinant protein was achieved (528.3 mg/L). Conclusion: With the optimization of these high cell density expression methods, reproducible manifold enhancement of soluble protein yields can be achieved for "difficult-to-express" GnRH-DFF40 compared to conventional expression methods.
Cell Cycle Modulation of CHO-K1 Cells Under Genistein Treatment Correlates with Cells Senescence, Apoptosis and ROS Level but in a Dose-Dependent Manner
Authors: Jenie Riris Istighfari , Amalina Nur Dina , Ilmawati Gagas Pradani Nur , Utomo Rohmad Yudi , Ikawati Muthi , Khumaira Annisa , Kato Jun- Ya , Meiyanto Edy
Published: August 2019

Purpose: Genistein, a soy isoflavone, exhibits a biphasic effect on cells proliferation with some different effects between ER-alpha and ER-beta. The objective of this present study is to determine the modulatory effect based on cell cycle progression under genistein treatment in combination with 17-β estradiol (E2) on CHO-K1 cells. Methods: The effect of genistein 0.1-100 µM on cells proliferation was examined by MTT assay. The modulation of genistein and estradiol (E2) on cell cycle and apoptosis were observed by using flowcytometry with PI and PI/AnnexinV staining, respectively. Moreover, the effect of genistein and E2 on senescence cells, and ROS level were determined by senescence-associated β-galactosidase (SA β-gal) staining and by using flowcytometry with 2’, 7’–dichlorofluorescin diacetate (DCFDA) staining, respectively. The expression level of the cell cycle and senescence protein markers were observed by immunoblotting. Results: Single treatment of genistein at physiologically achievable (low) concentration (<2 µM) induced proliferation of CHO-K1 cells while at a pharmacological (high) concentration (50 and 100 µM) suppressed cells proliferation. Interestingly, treatment of genistein at the physiological concentration in combination with E2 for 24, 48 and 72 h decreased cells viability on CHO-K1 cells compared to untreated cells. Further analysis of the cells showed that 50 µM genistein induced G2/M phase accumulation and induced apoptosis. Moreover, genistein induced cell senescence and increased ROS level. Immunoblotting analysis showed the decreasing of ERalpha, Bcl2, and ppRb protein level upon treatment of 1 µM Gen and 1 nM E2. Conclusion: Our results suggest that the cell proliferation inhibitory mechanism of genistein at pharmacological concentration involved the induction of cell senescence, and the elevation of ROS level. Moreover, the decreased of cells proliferation upon treatment of physiological concentration of genistein in combination with E2 may be correlated with the alteration of ER expression.