Resumo Como importante enzima, a xilanase é amplamente utilizada na indústria alimentícia, de celulose e têxtil. Diferentes aplicações de xilanase garantem condições específicas, incluindo temperatura e pH. Este estudo teve como objetivo realizar grânulos de alginato de sódio como carreador para imobilizar xilanase mutada relatada anteriormente de Neocallimastix patriciarum que expressa em E. coli, a atividade de imobilização da xilanase mutada foi elevada em cerca de 4% em pH 6 e 13% a 62 °C. Além disso, a xilanase mutada imobilizada reteve uma proporção maior de sua atividade do que o tipo amplo em termoestabilidade. Essas propriedades sugerem que a imobilização da xilanase mutada tem potencial para aplicação na indústria de biobranqueamento.

Abstract As an important enzyme, xylanase is widely used in the food, pulp, and textile industry. Different applications of xylanase warrant specific conditions including temperature and pH. This study aimed to carry out sodium alginate beads as carrier to immobilize previous reported mutated xylanase from Neocallimastix patriciarum which expressed in E. coli, the activity of immobilization of mutated xylanase was elevated about 4% at pH 6 and 13% at 62 °C. Moreover, the immobilized mutated xylanase retained a greater proportion of its activity than the wide type in thermostability. These properties suggested that the immobilization of mutated xylanase has potential to apply in biobleaching industry.

Resumo O objetivo deste estudo foi produzir alto rendimento de uma protease alcalina bacteriana local no sistema de leveduras, já que o envolvimento científico de microrganismos na produção de enzimas ainda não recebe atenção suficiente na Arábia Saudita. Amostras de solo foram coletadas da rizosfera de algumas plantas do deserto na Arábia Saudita. Noventa e três isolados bacterianos produtores de protease alcalina foram recuperados em ágar de leite desnatado a pH 9,4 e 45°C por 48 horas. O isolado D9 obtido da rizosfera de Heliotropium digynum na cidade de Dhahran foi o mais potente em relação à produtividade da enzima (184,6 U/ml). O gene completo da protease alcalina foi amplificado e apresentou o tamanho esperado (1300 pb). A análise de enzimas de restrição também verificou a integridade do produto de PCR. A sequência do gene da protease revelou uma fase de leitura aberta de 1329 nt, correspondendo ao comprimento total do gene da protease do isolado D9 que codifica uma proteína 443 aa. Após a ligação do gene amplificado pelo método de clonagem TA, a digestão com enzimas de restrição apropriadas confirmou a integridade do gene clonado. A inserção foi preparada por dois PCRs que foram conduzidos com um par de primers projetados especificamente para esta finalidade. O vetor de clonagem digerido e purificado pRS426/GAL1p-207-Glu-MS foi ligado com a inserção e então transformado em várias cepas de Saccharomyces cerevisiae por meio do método de eletroporação. A expressão máxima da protease foi feita por OS303 recombinante em meio contendo galactose (145,5 U/ml) com um aumento de aproximadamente duas vezes quando comparado com a cepa OS303 selvagem, e isso pode ser por causa da capacidade de ativar o operon gal.

Abstract The aim of this study was to produce high yield of a local bacterial alkaline protease in the yeast system because the scientific involvement of microorganisms in enzyme production is still not given enough attention in Saudi Arabia. Soil samples were collected from the rhizosphere of some desert plants in Saudi Arabia. Ninety-three alkaline protease producing bacterial isolates were recovered on skimmed-milk agar at pH 9.4 and 45°C for 48 hr. Isolate D9 obtained from the rhizosphere of Heliotropium digynum at Dhahran City was the most potent isolate in respect to enzyme productivity (184.6 U/ml). The full gene of alkaline protease was amplified and showed the expected size (1300 bp). Restriction enzymes analysis also verified the integrity of the PCR product. The sequence of the protease gene revealed an open reading frame of 1329 nt correspond to the full length of the protease gene of isolate D9 encoding a 443 aa protein. After ligation of the amplified gene by the TA cloning method, digestion with appropriate restriction enzymes confirmed the integrity of the cloned gene. The insert was prepared by two PCRs that were conducted with a pair of primers specifically designed for this purpose. The digested and purified cloning vector pRS426/GAL1p-207-Glu-MS was ligated with the insert then transformed into various strains of Saccharomyces cerevisiae via the electroporation method. Maximum protease expression was done by recombinant OS303 in galactose containing media (145.5 U/ml) with an approximately 2-fold increase when compared with the wild OS303 strain., this may be due to ability to activate gal operon.

The properties and deterioration in strength of cemented tailings backfill (CTB )in the underground acidic environment under oxidizing conditions were studied. X-ray diffraction analysis and scanning electron microscopy of the surface of the CTB was carried out, and the sulphuric acid corrosion mechanism elucidated. The properties tested included compressive strength, elastic modulus, cohesion, internal friction angle, variation in the hydrogen ion concentration, and stress-strain relationship in different corrosion periods. The damage model of the CTB was established considering the effects of parameters such as corrosion time and strain on the damage evolution. It was found that the compressive strength, elastic modulus, cohesion (binder effect), and internal friction angle increased at first and then decreased with exposure time. In the long term, the peak stress decreased with corrosion time while the peak strain increased; elastic modulus, and deformation modulus also decreased. The damage caused by corrosion and by load were related by means of a mathematical model, which revealed the relationships between sulphuric attack, load, and damage to backfill in complex underground environments. During corrosion, the expansion of gypsum and ettringite caused microfractures in the CTB. With increasing corrosion time, micro-cracks developed and proliferated. The mechanism of corrosion damage was found to be the dissolution of hydrogen ions and a sulphate ion reaction that produces an expansive substance, resulting in deterioration of the strength of CTB.

ABSTRACT Objective: To have anatomic measurements of carotid artery bifurcation (CAB) with 64-spiral computed tomography angiography (64-SCTA), and provide anatomic basis for related research. Methods: Imaging data of 92 subjects (45 males, 47 females, the age range 20–82 years and mean age 48.4 ± 6.1 years) without pathology of CAB, who underwent 64-SCTA in head and neck from June 1, 2008 to June 30, 2010, were selected from the Picture Archiving and Communication Systems in Zhongshan Hospital of Xiamen University, Fujian, China. On the 3D images, the angle and size of CAB were measured, and the statistical comparisons of measurements were made between the bilateral, sex and age groups. Results: The measurements of CAB were divided into young (≤ 40 years) and older (> 40 years) groups: bifurcation angle is 36.206° ± 10.210° and 49.343° ± 16.489°, respectively; the inner diameter of common carotid artery (CCA) is 6.820 ± 0.635 and 6.845 ± 0.838 mm, respectively; the proximal inner diameter of internal carotid artery (ICA) is 7.143 ± 0.992 and 7.476 ± 1.630 mm, of the enlargement is 7.568 ± 1.069 and 8.554 ± 1.733 mm, of the distal is 4.897 ± 0.508 and 5.123 ± 0.699 mm, respectively; the inner diameter of external carotid artery (ECA) is 4.324 ± 0.580 and 4.104 ± 0.638 mm, respectively. There were statistically significant differences in all the measurements between male and female groups, in the bifurcation angle, inner diameters of ICA and ECA between young and older groups, and in the bifurcation angle between the left and right (p < 0.05). Conclusion: A 64-SCTA with 3D image post-processing technique can clearly observe and show the CAB. All CAB measurements will provide the objective basis for applied anatomy, imaging diagnosis and surgery treatment.

Resumo As células epiteliais da mucosa atuam como a primeira barreira imunológica dos organismos e entram em contato direto com os patógenos. Portanto, os hospedeiros devem ter estratégias diferenciadas para combater os patógenos de forma eficiente. Trabalhos recentes revelaram que a oxidase dupla (DUOX) desempenha um papel fundamental para a imunidade da mucosa. A DUOX é uma proteína transmembrana geradora de espécies reativas de oxigênio (EROs) como seus principais produtos enzimáticos. Nesta revisão, apresentaremos as funções imunológicas da DUOX no trato respiratório e digestivo dos vertebrados.

Abstract Mucosal epithelial cells act as the first immunologic barrier of organisms, and contact directly with pathogens. Therefore, hosts must have differential strategies to combat pathogens efficiently. Reactive oxygen species (ROS), as a kind of oxidizing agents, participates in the early stage of killing pathogens quickly. Recent reports have revealed that dual oxidase (DUOX) plays a key role in mucosal immunity. And the DUOX is a transmembrane protein which produces ROS as their primary enzymatic products. This process is an important pattern for eliminating pathogens. In this review, we highlight the DUOX immunologic functions in the respiratory and digestive tract of vertebrates.

RESUMO O presente trabalho visa investigar os efeitos do extrato de semente de uva Proanthocyanidin (GSPE) sobre o desempenho da produção, o metabolismo e o status antioxidante de gado leiteiro Holstein em lactação precoce. Quarenta e oito vacas leiteiras multíparas Holstein foram divididas em quatro grupos (CON, G20, G40 e G80) e receberam 0, 20, 40 e 80mg de GSPE/kg de peso corporal/dia, respectivamente. O G20 aumentou significativamente o rendimento do leite em comparação com os outros grupos. A proteína e os sólidos não gordurosos do leite foram aumentados nos grupos G20, G40 e G80 somente no sétimo dia durante a experiência. Não foi observada diferença significativa na gordura do leite e na contagem de células somáticas, bem como nos parâmetros de metabolismo energético no sangue, na função hepática e na função renal entre os grupos em relação ao grupo controle. Não houve diferença significativa na glutationa peroxidase, na dimutase de superóxido, na capacidade antioxidante total e no peróxido de hidrogênio entre os grupos, mas o conteúdo malondialdeído do G20 aumentou significativamente no dia 14 em comparação com o CON, e tendia a aumentar no dia 28. Em conclusão, a alimentação de 20mg de GSPE/kg de peso corporal/dia foi associada a um aumento significativo no rendimento do leite, sem efeitos nocivos sobre a função hepática ou a renal, com o metabolismo de energia substancial e a melhoria dos parâmetros antioxidantes de gado leiteiro no início da lactação.

ABSTRACT Aims to investigate the effects of grape seed proanthocyanidin extract (GSPE) on production performance, metabolism, and anti-oxidative status of Holstein dairy cattle in early lactation. Forty-eight multiparous Holstein dairy cattle were assigned to four groups (CON, G20, G40 and G80) and supplied with 0, 20, 40, and 80mg GSPE/kg of body weight/day. G20 significantly increased milk yield compared with other groups. Milk protein and non-fat-solids were increased in G20, G40 and G80 groups compared with the control group only at the 7th day during the experiment. No significant difference was observed in milk fat and somatic cell count, nor on parameters of energy metabolism in blood, liver function and kidney function between the four groups. There was no significant difference in glutathione peroxidase, superoxide dismutase, total antioxidant capacity, and hydrogen peroxide between the groups; but the malondialdehyde content of G20 significantly increased at day 14 in comparison with CON, and tended to increase at the 28th day. In conclusion, feeding 20mg GSPE/kg of body weight/day was associated with a significant increase in milk yield without detrimental effects on liver or kidney function and with substantial energy metabolism and antioxidant parameters improvement in early lactation dairy cattle.

The majority of goats in Tanzania belong to the Small East African (SEA) breed, which exhibits large phenotypic variation. This study aimed to determine the genetic structure of, and relationships among four populations (Sukuma, Gogo, Sonjo, and Pare) of the SEA breed that have not been studied adequately. A total of 120 individuals (24 from each population) were analysed at eight microsatellite loci. In addition, 24 goats of the South African Boer breed were used as reference. Observed heterozygosity (Ho) ranged from 0.583 ± 0.04 for Sukuma to 0.659 ± 0.030 for Gogo, while expected heterozygosity (He) ranged from 0.632 ± 0.16 for Sukuma to 0.716 ± 0.16 for Boer. Five loci deviated from Hardy-Weinberg equilibrium (HWE) across populations. The mean number of alleles ranged from 4.75 ± 1.58 for Pare to 6.88 ± 3.00 for Sukuma. The mean inbreeding coefficient (F IS) ranged from 0.003 in Sonjo to 0.148 in Sukuma. The differentiation coefficient (F ST)was highest (0.085) between Boer and Sukuma and lowest (0.008) between Gogo and Sonjo. The largest genetic distance (0.456) was found between Sukuma and Boer, while the smallest (0.031) was between Gogo and Sonjo populations. Pare, Gogo, and Sonjo populations, formed one cluster, while Sukuma and Boer populations formed two separate clusters. From the findings, it can be concluded that the SEA goats in this study showed high in population genetic variation, which implies that there is good scope for their further improvement through selection within populations. The Sukuma population, which has fairly high inbreeding, is moderately differentiated from Pare, Sonjo, and Gogo goat populations, which showed a high level of admixture. Conservation and improvement strategies of the goats should be designed with first priority being on Sukuma goats.

Chronic heart failure (CHF) is a common chronic disease that requires much care. This study aimed to explore the effects of collaborative care model (CCM) on patients with CHF. A total of 114 CHF patients were enrolled in this study, and were randomly and equally divided into two groups: control and experimental. Patients in the two groups received either usual care or CCM for 3 continuous months. The impacts of CCM on the self-care ability and quality of life were assessed using self-care of heart failure index and short form health survey 12, respectively. Further, cardiac function was assessed by measuring left ventricular ejection fraction (LVEF) and the level of N-terminal pro-B-type natriuretic peptide (NT-proBNP), and by the 6-min walking test. Clinical and demographic characteristics of patients in the control and CCM groups were statistically equivalent. Compared with usual care, CCM significantly enhanced self-care abilities of patients with CHF, including self-care maintenance, self-care management and self-care confidence (all P<0.05). The physical and mental quality of life was also significantly improved by CCM (P<0.01 or P<0.05). Compared with usual care, CCM significantly increased the LVEF (P<0.01), decreased the NT-proBNP level (P<0.01), and enhanced exercise capacity (P<0.001). In conclusion, CCM improved the self-care, quality of life and cardiac function of patients with CHF compared with usual care.

Polymyositis (PM) is an autoimmune disease characterized by chronic inflammation in skeletal muscle. Mean platelet volume (MPV), a marker in the assessment of systemic inflammation, is easily measured by automatic blood count equipment. However, to our knowledge, there are no data in the literature with respect to MPV levels in PM patients. Therefore, in this study we aimed to investigate MPV levels in patients with PM. This study included 92 newly diagnosed PM patients and 100 healthy individuals. MPV levels were found to be significantly lower compared with healthy controls (10.3±1.23 vs 11.5±0.74 fL, P<0.001). Interestingly, MPV was found to be positively correlated with manual muscle test (MMT) score and negatively correlated with erythrocyte sedimentation rate (ESR) in patients with PM (r=0.239, P=0.022; r=−0.268, P=0.010, respectively). In addition, MPV was significantly lower in active PM patients compared with inactive PM patients (9.9±1.39 vs 10.6±0.92 fL, P=0.010). MPV was independently associated with PM in multivariate regression analyses, when controlling for hemoglobin and ESR (OR=0.312, P=0.031, 95%CI=0.108 to 0.899). The ROC curve analysis for MPV in estimating PM patients resulted in an area under the curve of 0.800, with sensitivity of 75.0% and specificity of 67.4%. Our results suggest that MPV is inversely correlated with disease activity in patients with PM. MPV might be a useful tool for rapid assessment of disease severity in PM patients.

Recent evidence shows that moxifloxacin could exert an antimicrobial effect against Helicobacter pylori in both in vitroand in vivo models. To systematically evaluate whether moxifloxacin-containing triple therapy could improve eradication rates and reduce side effects in first-line or second-line anti-H. pyloritreatment, eligible articles were identified by searches of electronic databases. We included all randomized trials comparing moxifloxacin-based triple therapy with standard triple or quadruple therapy during H. pylori eradication treatment. Statistical analysis was performed with Review Manager 5.0.10. Subanalysis/sensitivity analysis was also performed. We identified seven randomized trials (n=1263). Pooled H. pylori eradication rates were 79.03% (95%CI: 75.73-82.07) and 68.33% (95%CI: 64.44-72.04) for patients with moxifloxacin-based triple therapy or with standard triple or quadruple therapy, respectively (intention-to-treat analysis). The odds ratio (OR) was 1.82 (95%CI: 1.17-2.81), the occurrence of total side effects was 15.23% (95%CI: 12.58-18.20) and 27.17% (95%CI: 23.64-30.92) for groups with or without moxifloxacin, and the summary OR was 0.45 (95%CI: 0.26-0.77). In subgroup analyses, we noted that the second-line eradication rate in the moxifloxacin group was significantly higher than that in the quadruple therapy group (73.33 vs 60.17%, OR: 1.78, 95%CI: 1.16-2.73, P<0.001). However, there was no difference in first-line eradication treatment. Findings from this meta-analysis suggest that moxifloxacin-based triple therapy is more effective and better tolerated than standard triple or quadruple therapy. Therefore, a moxifloxacin-based triple regimen should be used in the second-line treatment of H. pylori infection.

The aim of the present study was to determine the effect of the combination of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and adriamycin (ADM) on the human breast cancer cell line MCF-7 and to identify potential mechanisms of apoptosis. Cell viability was analyzed by the MTT assay and the synergistic effect was assessed by the Webb coefficient. Apoptosis was quantified using the annexin V-FITC and propidium iodide staining flow cytometry. The mRNA expression of TRAIL receptors was measured by RT-PCR. Changes in the quantities of Bax and caspase-9 proteins were determined by Western blot. MCF-7 cells were relatively resistant to TRAIL (IC50 >10 µg/mL), while MCF-7 cells were sensitive to ADM (IC50 <10 µg/mL). A subtoxic concentration of ADM (0.5 µg/mL) combined with 0.1, 1, or 10 µg/mL TRAIL had a synergistic cytotoxic effect on MCF-7 cells, which was more marked with the combination of TRAIL (0.1 µg/mL) and ADM (0.5 µg/mL). In addition, the combined treatment with TRAIL and ADM significantly increased cell apoptosis from 9.8% (TRAIL) or 17% (ADM) to 38.7%, resulting in a synergistic apoptotic effect, which is proposed to be mediated by up-regulation of DR4 and DR5 mRNA expression and increased expression of Bax and caspase-9 proteins. These results suggest that the combination of TRAIL and ADM might be a promising therapy for breast cancer.

The selective separation of sulfapyridine (SPD) from sulfaquinoxaline (SQX) is investigated by applying high performance liquid chromatography (HPLC) with molecularly imprinted polymer (MIP) as the stationary phase. Herein we report the synthesis of a molecularly imprinting polymer, SPD-MIP, by free radical polymerization process. Because of this powerful method synthetic polymers with specific binding sites to template molecule are provided. In addition, the separation performances were represented by using buffer/acetonitrile (3/2, v/v) as mobile phase under 272 nm UV detection. In order to compare the chromatographic data from the stationary phase, capacity factor (k') and separation factors (á) were given. The value of 2.71 (á) revealed that the MIP was able to recognize structurally subtle differences from the template molecule. Our results are discussed with regard to the amount of template, the composition of the chromatographic mobile phase and adsorption capacity.

The objective of the present study was to investigate the effects of recombinant human growth hormone (rhGH) on the intestinal mucosa barrier of septic rats and explore its possible mechanism. Female Sprague-Dawley rats were randomized into three groups: control, Escherichia coli-induced sepsis (S) and treatment (T) groups. Groups S and T were subdivided into subgroups 1d and 3d, respectively. Expression of liver insulin-like growth factor-1 (IGF-1) mRNA, Bcl-2 and Bax protein levels and the intestinal Bax/Bcl-2 ratio, and plasma GH and IGF-1 levels were determined. Histological examination of the intestine was performed and bacterial translocation was determined. rhGH significantly attenuated intestinal mucosal injuries and bacterial translocation in septic rats, markedly decreased Bax protein levels, inhibited the decrease of Bcl-2 protein expression and maintained the Bax/Bcl-2 ratio in the intestine. rhGH given after sepsis significantly improved levels of plasma GH (T1d: 1.28 ± 0.24; T3d: 2.14 ± 0.48 µg/L vs S1d: 0.74 ± 0.12; S3d: 0.60 ± 0.18 µg/L; P < 0.05) and IGF-1 (T1d: 168.94 ± 65.67; T3d: 201.56 ± 64.98 µg/L vs S1d: 116.72 ± 13.96; S3d: 107.50 ± 23.53 µg/L; P < 0.05) and expression of liver IGF-1 mRNA (T1d: 0.98 ± 0.20; T3d: 1.76 ± 0.17 vs S1d: 0.38 ± 0.09; S3d: 0.46 ± 0.10; P < 0.05). These findings indicate that treatment with rhGH had beneficial effects on the maintenance of the integrity of the intestinal mucosa barrier in septic rats.