Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.

Physical training prevents body weight gain but does not modify adipose tissue gene expression

The relationship of body weight (BW) with white adipose tissue (WAT) mass and WAT gene expression pattern was investigated in mice submitted to physical training (PT). Adult male C57BL/6 mice were submitted to two 1.5-h daily swimming sessions (T, N = 18), 5 days/week for 4 weeks or maintained sedentary (S, N = 15). Citrate synthase activity increased significantly in the T group (P < 0.05). S mice had a substantial weight gain compared to T mice (4.06 ± 0.43 vs 0.38 ± 0.28 g, P < 0.01). WAT mass, adipocyte size, and the weights of gastrocnemius and soleus muscles, lung, kidney, and adrenal gland were not different. Liver and heart were larger and the spleen was smaller in T compared to S mice (P < 0.05). Food intake was higher in T than S mice (4.7 ± 0.2 vs 4.0 ± 0.3 g/animal, P < 0.05) but oxygen consumption at rest did not differ between groups. T animals showed higher serum leptin concentration compared to S animals (6.37 ± 0.5 vs 3.11 ± 0.12 ng/mL). WAT gene expression pattern obtained by transcription factor adipocyte determination and differentiation-dependent factor 1, fatty acid synthase, malic enzyme, hormone-sensitive lipase, adipocyte lipid binding protein, leptin, and adiponectin did not differ significantly between groups. Collectively, our results showed that PT prevents BW gain and maintains WAT mass due to an increase in food intake and unchanged resting metabolic rate. These responses are closely related to unchanged WAT gene expression patterns.

Rhein induces apoptosis of human gastric cancer SGC-7901 cells via an intrinsic mitochondrial pathway

Rhein is a primary anthraquinone found in the roots of a traditional Chinese herb, rhubarb, and has been shown to have some anticancer effects. The aim of the present study was to investigate the effect of rhein on the apoptosis of the human gastric cancer line SGC-7901 and to identify the mechanism involved. SGC-7901 cells were cultured and treated with rhein (0, 50, 100, 150, and 200 µM) for 24, 48, or 72 h. Relative cell viability assessed by the MTT assay after treatment was 100, 99, 85, 79, 63% for 24 h; 100, 98, 80, 51, 37% for 48 h, and 100, 97, 60, 36, 15% for 72 h, respectively. Cell apoptosis was detected with TUNEL staining and quantified with flow cytometry using annexin FITC-PI staining at 48 h after 100, 200 and 300 µm rhein. The percentage of apoptotic cells was 7.3, 21.9, 43.5%, respectively. We also measured the mRNA levels of caspase-3 and -9 using real-time PCR. Treatment with 100 µM rhein for 48 h significantly increased mRNA expression of caspase-3 and -9. The levels of apoptosis-related proteins including Bcl-2, Bax, Bcl-xL, and pro-caspase-3 were evaluated in rhein-treated cells. Rhein increased the Bax:Bcl-2 ratio but decreased the protein levels of Bcl-xL and pro-caspase-3. Moreover, rhein significantly increased the expression of cytochrome c and apoptotic protease activating factor 1, two critical components involved in mitochondrial pathway-mediated apoptosis. We conclude that rhein inhibits SGC-7901 proliferation by inducing apoptosis and this antitumor effect of rhein is mediated in part by an intrinsic mitochondrial pathway.

Capecitabine maintenance therapy in patients with recurrent or metastatic breast cancer

Our objective was to investigate the efficacy and safety of capecitabine maintenance therapy (CMT) after capecitabine-based combination chemotherapy in patients with metastatic breast cancer. The clinical data of 139 metastatic breast cancer patients treated from March 2008 to May 2012 with capecitabine-based combination chemotherapy were retrospectively analyzed. When initial disease control was achieved by the combination chemotherapy, we used CMT for 50 patients, while 37 patients were treated with a different (non-CMT) maintenance therapy. We compared time to progression (TTP), objective response rate, disease control rate, clinical benefit rate, and safety of the two groups, and a sub-group analysis was performed according to pathological characteristics. Sixty-four percent of the patients received a median of six cycles of a docetaxel+capecitabine combination chemotherapy regimen (range 1-45); the median TTP (MTTP) for the complete treatment was 9.43 months (95%CI=8.38-10.48 months) for the CMT group and 4.5 months (95%CI=4.22-4.78 months; P=0.004) for the non-CMT group. The MTTPs for the maintenance therapies administered after the initial capecitabine combined chemotherapy were 4.11 months (95%CI=3.34-4.87 months) for the CMT group and 2.0 months (95%CI=1.63-2.38 months) for the non-CMT group. Gastrointestinal side effects, decreased white blood cells and palmar-plantar erythrodysesthesia were the main adverse reactions experienced with the combination chemotherapies, CMT and non-CMT treatments. No significant differences in the incidence of adverse reactions were detected in the CMT and non-CMT patients. After initial disease control was achieved with the capecitabine-based combination chemotherapy, CMT can significantly prolong TTP rates with a favorable safety profile.

Aeromonas spp. e Plesiomonas shigelloides isoladas a partir de mexilhões (Perna perna) in natura e pré-cozidos no Rio de Janeiro, RJ

O ecossistema aquático é o habitat de mexilhões (Perna perna), animais filtradores que refletem a qualidade ambiental através de análise microbiológica de sua carne. No presente trabalho avaliou-se a presença de patógenos emergentes (Aeromonas hydrophila e Plesiomonas shigelloides), em mexilhões in natura e pré-cozidos coletados por pescadores da Estação Experimental de Cultivo de Mexilhões situada em Jurujuba, Niterói, Rio de Janeiro. Foram analisadas 86 amostras de mexilhões (43 in natura e 43 pré-cozidos) as quais foram submetidas a enriquecimento em Água Peptonada Alcalina (APA) acrescida de 1 e 3% de Cloreto de Sódio (NaCl) e em solução Salina de Butterfield, incubadas a 37ºC por 24 horas. Em seguida, foram semeadas em Ágar Seletivo para Pseudomonas-Aeromonas (GSP), Ágar Tiossulfato Citrato Bile Sacarose (TCBS) e Ágar Inositol Bile Verde Brilhante (IBB). A análise geral dos resultados permitiu a identificação de Areomonas spp e Plesiomonas shigelloides em 86% das amostras de mexilhões in natura e pré-cozidas avaliadas. A posterior caracterização bioquímica permitiu a identificação das espécies Aeromonas media (37,10%), A. hydrophila (15,50%), A. caviae (14,80%), A. veronii biogrupo veronii (11,60%), Aeromonas sp. (7,36%), A. sobria (4,20%), A. trota (4,20%), A. schubertii (1,31%), A. jandaei (1,31%), A. veronii biogrupo sobria (0,52%) e Plesiomonas shigelloides (2,10%). A relevância epidemiológica desses microrganismos em casos de gastrenterite humana, após consumo de mexilhões crus ou parcialmente cozidos, revela a importância de alertar as autoridades de Saúde Pública no Brasil, sobre a presença desses patógenos na cadeia alimentar e seus riscos para a saúde humana.

"Iogurte" de soja suplementado com oligofrutose e inulina

A população de um modo geral está mais consciente da relação existente entre alimentação e saúde. Sendo assim, a indústria busca novas alternativas para o desenvolvimento de alimentos de boa aceitabilidade e com ingredientes capazes de promover a saúde. A soja e prebióticos como oligofrutose e inulina possuem grande potencial de aplicação na indústria, devido as suas importantes propriedades funcionais. O objetivo deste trabalho foi determinar, utilizando a metodologia da superfície de resposta, as melhores condições para o desenvolvimento de um "iogurte" de soja suplementado com oligofrutose e inulina. Utilizou-se o delineamento fatorial incompleto 3³, tendo oligofrutose (%), inulina (%) e tempo de fermentação (h) como variáveis independentes e pH e acidez titulável como respostas. A análise de regressão mostrou que os modelos obtidos foram preditivos. A otimização conjunta bilateral apontou a formulação contendo 14,24% de oligofrutose, 4,43% de inulina e 6 horas como as melhores condições para o preparo de "iogurte" de soja. Essa formulação apresentou pH 4,6; acidez 0,37%; 16,2% de carboidratos; 2,01% de lipídeos; 3,54% de proteínas; 0,40% de cinzas e 77,85% de umidade. O teor de cálcio, ferro e sódio foram de 37; 0,99 e 15mg/100g, respectivamente. Pode-se afirmar que oligofrutose e inulina podem ser utilizadas na suplementação de "iogurte" de soja, constituindo um produto com ingredientes potencialmente funcionais.

Avaliação físico-química de filés de tilápia (Oreochromis niloticus) submetidos à sanitização

No presente trabalho foi avaliado o efeito dos agentes sanificantes dicloroisocianurato de sódio na concentração de 3 a 3,5 mg.L-1 (T1), dicloroisocianurato de sódio associado ao ultra-som (T2), ozônio na concentração de 3 a 3,5 mg.L-1 (T3), ozônio associado ao ultra-som (T4) e controle (T5) sobre a composição centesimal, perfil de ácidos graxos (AG), teor de colesterol e índice de peróxido de filés de tilápia. Os tratamentos afetaram (p < 0,05) a composição centesimal. Os filés foram tratados por imersão, durante 20 minutos, em soluções que apresentavam pH 6,0 e temperatura de 5 °C. Os sanificantes causaram uma perda de água e aumentaram a concentração de proteínas no filé, tendo esse fato sido notado mais acentuadamente no T3 (77,16% de umidade e 19,73% de proteína) quando comparado ao T5 (78,60% de umidade e 18,71% de proteína). Foi demonstrada diferença significativa (p < 0,05) na fração lipídica para ácidos graxos saturados (AGS), ácidos graxos monoinsaturados (AGM) e ácidos graxos polinsaturados (AGP) entre os tratamentos propostos. Os AGS apresentaram diferença entre os T3, T4 e T5, com as respectivas médias 37, 33,50 e 36,30%. Os T1 e T2 (34,90 e 34,80%) não diferiram entre si, mas foram diferentes dos T3, T4 e T5 (39,30, 36,30 e 39,10%) para AGM. O T3 foi o que mais preservou a fração AGM. Os AGP não apresentaram diferença entre os T1 e T2 (23 e 22,50%) e os grupos T3, T4 e T5 (18,30, 20,40 e 17,70%) diferiram entre si, tendo o T1 apresentado as maiores médias. Obteve-se uma redução mais acentuada do nível de colesterol no grupo T2 (43,94 mg.100 g-1) quando comparado com T5 (54,77 mg.100 g-1). Os teores médios de peróxido encontrados nos filés submetidos aos tratamentos não revelaram diferença estatística segundo análise de variância (p > 0,05), apesar dos valores serem bastante diferentes, apresentando uma variação de 6,07 a 39,57 meq.kg-1.

Sensory evaluation of rice fortified with iron

The objective of this study was to examine sensory differences between conventional rice and iron-fortified Ultra Rice rice (UR) and determine consumer acceptance. Differences between both types of rice were analyzed using the Duo-Trio Test on 37 non-trained judges. The Acceptance Test evaluated general rice appearance, color, aroma and taste by 43 non-trained judges, using a 7-point hedonic scale with extremes ranging from "really disliked" and "really liked." There were no significant differences between the analyzed samples of conventional rice and UR rice using Duo-Trio Test. Mean values from the Acceptance Test for the evaluated attributes (general appearance, color, aroma e taste) were 5.6 and 5.9, corresponding to "liked" and "really liked" according to the hedonic scale, respectively. The iron did not alter the sensory characteristics of the final product, and the iron-fortified rice was well accepted.