Nutritional value, total dry matter losses, effluent production and pollutant potential in Brachiaria brizantha cv. Paiaguás grass / [Valor nutricional, perdas totais de matéria seca, produção e potencial poluidor de efluentes em silagem de Brachiaria brizantha cv. Paiaguás]

The objective of the present study was to evaluate losses, production and polluting potential of the effluent, nutritional value and aerobic stability of silages of Brachiaria brizantha cv. Paiaguás grass, in different particle sizes and compaction density in silage. Three theoretical particle sizes (TTP 5; 8 and 12mm) and three compaction densities (DC 550; 600 and 650kg/m3) were evaluated, distributed in a factorial design (3 x 3), with four repetitions. The highest volume of effluent was found in silages with higher compaction densities (600 and 650kg/m3) and lower TTP (5 and 8mm). The highest chemical oxygen demand and biochemical oxygen demand were registered in the treatment with TTP of 5mm and higher DC (600 and 650kg/m3). Greater in vitro digestibility of DM was verified in the silage chopped at 5 and 8mm. There was no break in aerobic stability for 216 hours. Silage with a low compaction density 550kg/m3 and processing with a theoretical particle size of 12mm reduces effluent losses. In general, the nutritional value of Paiaguás grass was not influenced by the treatments. Different particle sizes and compaction density did not change the aerobic stability of silages.(AU)

Objetivou-se avaliar perdas, produção e potencial poluidor do efluente, valor nutricional e estabilidade aeróbia de silagens do capim Brachiaria brizantha cv. Paiaguás, em diferentes tamanhos de partícula e densidade de compactação na ensilagem. Foram avaliados três tamanhos teóricos de partícula (TTP 5; 8 e 12mm) e três densidades de compactação (DC 550; 600 e 650kg/m3), distribuídos em arranjo fatorial (3 x 3), com quatro repetições. O maior volume de efluente foi verificado nas silagens com maiores densidades de compactação (600 e 650kg/m3) e menores TTP (5 e 8mm). As maiores demanda química de oxigênio e demanda bioquímica de oxigênio foram registradas no tratamento com TTP de 5mm e nas maiores DC (600 e 650kg/m3). Maior digestibilidade in vitro da MS (média de 57,2%) foi verificada na silagem picada a 5 e 8mm. Não houve quebra da estabilidade aeróbia durante 216 horas. A ensilagem com baixa densidade de compactação (550kg/m3) e o processamento com tamanho teórico de partículas 12mm reduzem as perdas por efluente. O valor nutricional da silagem de capim-paiaguás, em geral, não foi influenciado pelos tratamentos. Diferentes tamanhos de partícula e densidade de compactação não alteraram a estabilidade aeróbia das silagens.(AU)

Regulatory control of the Na-Cl co-transporter NCC and its therapeutic potential for hypertension

Hypertension is the largest risk factor for cardiovascular disease, the leading cause of mortality worldwide. As blood pressure regulation is influenced by multiple physiological systems, hypertension cannot be attributed to a single identifiable etiology. Three decades of research into Mendelian forms of hypertension implicated alterations in the renal tubular sodium handling, particularly the distal convoluted tubule (DCT)-native, thiazide-sensitive Na-Cl cotransporter (NCC). Altered functions of the NCC have shown to have profound effects on blood pressure regulation as illustrated by the over activation and inactivation of the NCC in Gordon's and Gitelman syndromes respectively. Substantial progress has uncovered multiple factors that affect the expression and activity of the NCC. In particular, NCC activity is controlled by phosphorylation/dephosphorylation, and NCC expression is facilitated by glycosylation and negatively regulated by ubiquitination. Studies have even found parvalbumin to be an unexpected regulator of the NCC. In recent years, there have been considerable advances in our understanding of NCC control mechanisms, particularly

Multi-Epitope Peptide-Based and VacciniaBased Universal Influenza Vaccine Candidates Subjected to Clinical Trials

@#In light of the limited protection conferred by current influenza vaccines, immunisation using universal influenza vaccines has been proposed for protection against all or most influenza sub-types. The fundamental principle of universal influenza vaccines is based on conserved antigens found in most influenza strains, such as matrix 2, nucleocapsid, matrix 1 and stem of hemagglutinin proteins. These antigens trigger cross-protective immunity against different influenza strains. Many researchers have attempted to produce the conserved epitopes of these antigens in the form of peptides in the hope of generating universal influenza vaccine candidates that can broadly induce cross-reactive protection against influenza viral infections. However, peptide vaccines are poorly immunogenic when applied individually owing to their small molecular sizes. Hence, strategies, such as combining peptides as multi-epitope vaccines or presenting peptides on vaccinia virus particles, are employed. This review discusses the clinical and laboratory findings of several multi-epitope peptide vaccine candidates and vaccinia-based peptide vaccines. The majority of these vaccine candidates have reached the clinical trial phase. The findings in this study will indeed shed light on the applicability of universal influenza vaccines to prevent seasonal and pandemic influenza outbreaks in the near future.

Nutritive Parameters Evolution of Maize Seeds Conserved by Triple Bagging System and Biopesticides (Lippia multiflora and Hyptis suaveolens Leaves) in Cote d’Ivoire

This study, initiated in Côte d'Ivoire, aimed to evaluate the effectiveness of the triple bagging system associated or not with biopesticides on the conservation of biochemical parameters, in particular its nutritional potential according to a central composite design (CCD). It was carried in Côte d'Ivoire at Laboratory of Biochemistry and Food Science from March 2016 to September 2017. Shelf life, biopesticides rate and interactions between shelf life and biopesticides had a significant influence on the biochemical characteristics of maize. The polypropylene bag (control) had the highest values after eighteen (18) months of moisture storage (9.02% to 16.99%) and showed very high fibre losses (P<0.001) (5.78% to 4.28%), total sugars (2.62% to 1.30%), reducing sugars (0.47% to 0.27%), starch (75.20% to 46.10%), fat (5.51% to 3.33%), protein (8.60% to 6.87%), total carbohydrate (75.20% to 71.51%), ash (1.68% to 1.30%) and energy value (384.78% to 343.48%). Concerning the triple bagging system without biopesticides, the variation is similar to the treatments that received the biopesticides up to 9.5 months of storage before presenting values almost similar to the control bag after the 18 months of storage. While triple bagging systems with the presence of biopesticides after 18 months of storage show slight variations in moisture (9.02% to 12.47%), fibre (5.78% to 5.56%), total sugars (2.62% to 1,88%), reducing sugars (0.47% to 0.37%), starch (75.20% to 60.03%), fat (5.51% to 5.00%), protein (8.60% to 7.84%), total carbohydrates (75.20% to 72.69%), ash (1.68% to 1.50%) and energy value (384.78% to 368.93%). The results of these tests show that maize grains stored in the presence of biopesticides best retain their biochemical characteristics. Also, the results indicate that the rate of 1.01% biopesticides could be recommended for maintaining all biochemical parameters up to 18 months of storage.

Eukaryotic Multi-subunit DNA dependent RNA Polymerases: An Insight into Their Active Sites and Catalytic Mechanism

Aim:To analyze the most complex multi-subunit (MSU) DNA dependent RNA polymerases (RNAPs) of eukaryotic organisms and find out conserved motifs, metal binding sites and catalytic regions and propose a plausible mechanism of action for these complex eukaryoticMSU RNAPs, using yeast (Saccharomyces cerevisiae) RNAP II, as a model enzyme.Study Design: Bioinformatics, Biochemical, Site-directed mutagenesis and X-ray crystallographic data were analyzed.Place and Duration of Study: School of Biotechnology, MaduraiKamaraj University, Madurai, India, between 2007-2013. Methodology:Bioinformatics, Biochemical, Site-directed mutagenesis (SDM) and X-ray crystallographic data of the enzyme were analyzed. The advanced version of Clustal Omega was used for protein sequence analysis of the MSU DNA dependent RNAPs from various eukaryotic sources. Along with the conserved motifs identified by the bioinformatics analysis, the data already available by biochemical and SDM experiments and X-ray crystallographic analysis of these enzymes were used to confirm the possible amino acids involved in the active sites and catalysis. Results:Multiple sequence alignment (MSA) of RNAPs from different eukaryotic organisms showed a large number of highly conserved motifs among them. Possible catalytic regions in the catalytic subunits of the yeast Rpb2 (= β in eubacteria) and Rpb1 (= β’ in eubacteria) consist of an absolutely conserved amino acid R, in contrast to a K that was reported for DNA polymerases and single subunit (SSU) RNAPs. However, the invariant ‘gatekeeper/DNA template binding’ YG pair that was reported in all SSU RNAPs, prokaryotic MSU RNAPs and DNA polymerases is also highly conserved in eukaryotic Rpb2 initiation subunits, but unusually a KG pair is found in higher eukaryotes including the human RNAPs. Like the eubacterial initiation subunits of MSU RNAPs, the eukaryotic initiation subunits, viz. Rpb2, exhibit very similar active site and catalytic regions but slightly different distance conservations between the templatebinding YG/KG pair and the catalytic R. In the eukaryotic initiation subunits, the proposed catalytic R is placed at the -9thposition from the YG/KG pair and an invariant R is placed at -5 which are implicated to play a role in nucleoside triphosphate (NTP) selection as reported for SSU RNAPs (viral family) and DNA polymerases. Similarly, the eukaryotic elongation subunits (Rpb1) are also found to be very much homologous to the elongation subunits (β’) of prokaryotes. Interestingly, the catalytic regionsare highly conserved, and the metal binding sites are absolutely conserved as in prokaryotic MSU RNAPs. In eukaryotes, the template binding YG pair is replaced with an FG pair. Another interesting observation is, similar to the prokaryotic β’ subunits, inthe eukaryotic Rpb1 elongation subunits also, the proposed catalytic R is placed double the distance, i.e., -18 amino acids downstream from the FG pair unlike in the SSU RNAPs and DNA polymerases where the distance is only -8 amino acids downstream from the YG pair. Thus, the completely conserved FG pair, catalytic R with an invariant R, at -6thposition are proposed to play a crucial role in template binding, NTP selection and polymerization reactions in the elongation subunits of eukaryotic MSU RNAPs. Moreover, the Zn binding motif with the three completely conserved Cs is also highly conserved in the eukaryotic elongation subunits. Another important difference is that the catalytic region is placed very close to the N-terminal region in eukaryotes.Conclusions: Unlike reported for the DNA polymerases and SSU RNA polymerases, the of eukaryotic MSU RNAPs use an R as the catalytic amino acid and exhibit a different distance conservation in the initiation and elongation subunits. An invariant Zn2+binding motif found in the Rpb1 elongation subunits is proposed to participate in proof-reading function. Differences in the active sites of bacterial and human RNA polymerases may pave the way for the design of new and effective drugs for many bacterial infections, including the multidrug resistant strains which are a global crisis at present

Inducible cellulase production from an organic solvent tolerant Bacillus sp. SV1 and evolutionary divergence of endoglucanase in different species of the genus Bacillus

Abstract Bacteria are important sources of cellulases with various industrial and biotechnological applications. In view of this, a non-hemolytic bacterial strain, tolerant to various environmental pollutants (heavy metals and organic solvents), showing high cellulolytic index (7.89) was isolated from cattle shed soil and identified as Bacillus sp. SV1 (99.27% pairwise similarity with Bacillus korlensis). Extracellular cellulases showed the presence of endoglucanase, total cellulase and β-glucosidase activities. Cellulase production was induced in presence of cellulose (3.3 times CMCase, 2.9 times FPase and 2.1 times β-glucosidase), and enhanced (115.1% CMCase) by low-cost corn steep solids. An in silico investigation of endoglucanase (EC 3.2.1.4) protein sequences of three Bacillus spp. as query, revealed their similarities with members of nine bacterial phyla and to Eukaryota (represented by Arthropoda and Nematoda), and also highlighted of a convergent and divergent evolution from other enzymes of different substrate [(1,3)-linked beta-d-glucans, xylan and chitosan] specificities. Characteristic conserved signature indels were observed among members of Actinobacteria (7 aa insert) and Firmicutes (9 aa insert) that served as a potential tool in support of their relatedness in phylogenetic trees.

Establishment and application of a TaqMan real-time fluorescence quantitative PCR for detection of tree shrew adenovirus(TAV) / 中国比较医学杂志

Objective To establish a quick and accurate method for detection of tree shrew adenovirus(TAV) using TaqMan real-time fluorescence quantitative PCR. Methods Based on the published TAV genome sequence, a 3' conserved sequence was used to design specific probe primers. A standard curve was prepared using a recombinant plasmid containing the target gene fragment. A real-time fluorescence quantitative PCR method was established for detecting TAV based on TaqMan probe. Results The detection method was specific and was not cross-reactive with other common pathogens. The detection limit of the method was 3.7 copies/μL,showing a high sensitivity. The correlation coefficient was 0.998, and the efficiency was 95.7%. The amplification result showed a fine linear relationship,and the repeatability test effect was good. Conclusions The TAV real-time quantitative PCR detection method based on TaqMan probe has been successfully established. It has high sensitivity and reproducibility and can be applied to early detection of TAV infection.

The importance of GDN motif in domain III and GXGXG motif in domain VI of L protein in replication of Nipah virus

Abstracts@#This study aims to determine the importance of conserved GDN motif in domain III and GXGXG motif in domain VI in Nipah virus (NiV) L protein. Four mutated L genes produced in an earlier study were inserted individually into plasmid pCITE. Optimised transfection protocol was successful in transfecting these plasmids, two helper plasmids (coding for N and P protein), NiV minigenome containing chloramphenicol acetyltransferase (CAT) reporter gene and T7 promoter. Successful in vitro transcription/translation in the NiV minireplicon system was monitored by CAT expression. In conclusion, GXGXG motif was important in the NiV minireplicon system but change of GDN motif does not affect L protein.

Características produtivas e nutricionais do feno de trigo cultivado em dois níveis de adubação nitrogenada e estádios de colheita / Productive and nutritional characteristics of wheat hay cultivated in two nitrogen fertilization levels and harvest stages

O objetivo do presente trabalho foi avaliar a produção de fitomassa fresca e de fitomassa seca, assim como a composição física e bromatológica do feno do trigo cultivar BRS Umbu sob efeitos de dois níveis de adubação nitrogenada (120kg ha-1 e 180kg ha-1) e dois estádios fenológicos de colheita (pré-florescimento e grão farináceo). O delineamento experimental foi blocos ao acaso, em esquema fatorial 2x2, composto por quatro tratamentos com quatro repetições. Não houve interação (P>0,05) entre os níveis de adubação nitrogenada e os estádios fenológicos de colheita para todas as variáveis. A produção de fitomassa seca (P<0,05) nos estádios de grão farináceo e pré-florescimento foi de 10.171 e 4.982kg ha-1, respectivamente. A maior dose de N incrementou a produção de fitomassa seca em 775kg ha-1. Houve aumento da participação de espigas com o avanço do ciclo, apresentando-se 43,0% e 16,2% nos estádios grão farináceo e pré-florescimento, respectivamente. No estádio fenológico de pré-florescimento, a participação de folha verde foi superior (37,1% contra 9,8% da MS total). O feno colhido em estádio de grão farináceo apresentou menores teores de proteína bruta e fibra em detergente neutro. Os fenos produzidos apresentaram características distintas, o que permite seu uso em diferentes estratégias de alimentação de ruminantes.(AU)

The objective was to evaluate the production of the fresh and dry weight, physical and chemical composition of wheat hay, cv. BRS UMBU, under effects of two levels of nitrogen fertilization (120kg ha-1 and 180kg ha-1) and two harvest stages (pre-flowering and grain dough). The experimental design was randomized blocks in factorial 2X2, composed of four treatments with four replications. There was no interaction (p>0.05) between the levels of nitrogen fertilization and growth stages of harvest for all variables. The production of dry matter in the dough stage and pre-flowering were 10.171 and 4.982kg ha-1, respectively. The higher N rates increased the production of dry matter of 775kg ha-1. There was increased participation of spikes with the advancement of the cycle, presenting 43.0% and 16.2% in the dough stage and pre-flowering, respectively. In the growth stage of pre-flowering, the share of green leaf was higher (37.1% against 9.8% of the total MS). The hay harvested at dough stage had lower NDF and CP levels. Thus, each treatment presented favorable characteristics allowing its use in different strategies in ruminant nutrition.(AU)

Research progress on the type Ⅸ secretion system of Porphyromonas gingivalis / 华西口腔医学杂志

In recent years, the study found that Porphyromonas gingivalis type Ⅸ secretion system (T9SS) is a novel protein secretion system, also known as Por secretion system (PorSS). Unlike the eight protein secretion systems found in the past, the system is a polyprotein complex found only in Bacteroides. The secreted proteins have both N- and C-terminus, where the former includes Sec-dependent type Ⅰ signals peptide, and the latter contains conserved domains (C-terminal conserved domain, CTD). Porphyromonas gingivalis T9SS includes proteins such as intima, outer membrane, cytoplasm, and cell cycle, including at least 34 proteins containing CTD. Porphyromonas gingivalis T9SS is involved in regulating associated virulence factors including gingivin, fimbriae, lipopolysaccharide, HBP35, CPG70 protein and peptidyl-arginine deiminase. These CTD-containing virulence proteins are localized by T9SS and then released to the extracellular domain, thereby destroying periodontal tissue. Therefore, this review summarizes the research progress on the T9SS of Porphyromonas gingivalis.

Genomic Analysis of miR-21-3p and Expression Pattern with Target Gene in Olive Flounder

MicroRNAs (miRNAs) act as regulators of gene expression by binding to the 3’ untranslated region (UTR) of target genes. They perform important biological functions in the various species. Among many miRNAs, miR-21-3p is known to serve vital functions in development and apoptosis in olive flounder. Using genomic and bioinformatic tools, evolutionary conservation of miR-21-3p was examined in various species, and expression pattern was analyzed in olive flounder. Conserved sequences (5’-CAGUCG-3’) in numerous species were detected through the stem-loop structure of miR-21-3p. Thus, we analyzed target genes of miR-21-3p. Among them, 3’ UTR region of PPIL2 gene indicated the highest binding affinity with miR-21-3p based on the minimum free energy value. The PPIL2 gene showed high expression levels in testis tissue of the olive flounder, whereas miR-21-3p showed rather ubiquitous expression patterns except in testis tissue, indicating that miR-21-3p seems to control the PPIL2 gene expression in a complementary repression manner in various tissues of olive flounder. Taken together, this current study contributes to infer the target gene candidates for the miR-21-3p using bioinformatics tools. Furthermore, our data offers important information on the relationship between miR-21-3p and target gene for further functional study.

The lipid droplet: A conserved cellular organelle

The lipid droplet (LD) is a unique multi-functional organelle that contains a neutral lipid core covered with a phospholipid monolayer membrane. The LDs have been found in almost all organisms from bacteria to humans with similar shape. Several conserved functions of LDs have been revealed by recent studies, including lipid metabolism and trafficking, as well as nucleic acid binding and protection. We summarized these findings and proposed a hypothesis that the LD is a conserved organelle.

A purple non-sulfur bacterium producing polyhydroxybutyrate and the conserved region of pha synthase gene / Bactéria púrpura não sulfurosa produtora de polihidroxibutirato e a região conservada do gene pha sintase

This study aimed to screen purple non-sulfur bacteria capable of accumulating granules or polyhydroxybutyrate (PHB) inside the cells, identify the potent strain, assay the enzyme or PHA synthase, and compare the PHB synthase gene with that of related strains. A total of 58 strains of purple non-sulfur bacteria were isolated from 108 samples of chicken feces in the chicken-egg farm of the Department of Animal Science, Faculty of Natural Resources at Prince of Songkla University, Hat Yai, Thailand. After cultivating the bacteria in glutamate malate (GM) medium without added glutamic acid under light (3,000 Lux) at 35oC for 5 days, the intracellular biopolymer granules of the bacteria were observed by using a Confocal Laser Scanning Microscope (CLSM) with excitation and emission wavelength of 530 and 605 nm, respectively. Gas chromatography (GC) was carried out for quantitative analysis of PHB. There were five strains, CH12, CH52, CH72, CH90 and CH92, showed biopolymer granules under CLSM, and accumulated PHB 5, 1.7, 1.5, 1.4 and 1.8% (w w-1) of the cell dry weight (CDW), respectively. The 16S rDNA sequence analysis of CH12 strain showed a high homology of 100% correlation to that of Rhodopseudomonas palustris strain NCIB8288. Regarding the taxonomic characteristics and 16S rDNA sequence analysis, CH12 strain was identified as Rps. palustris NCIB8288. The PHA synthase activity of the crude extract from CH12 strain was 25 units/mL. The conserved regions could be aligned and selected among 5 strains of Rhodopseudomonas palustris (strains BisA53, TIE-1, CGA009, HaA2 and BisB18). The purified PCR product was obtained for further studies.

Este estudo teve como objetivo rastrear bactérias púrpuras não sulfurosas capazes de acumular grânulos ou polihidroxibutirato (PHB) dentro das células, identificar a estirpe potente, ensaiar a enzima ou PHA sintaxe, e comparar com o gene PHB sintase com aquele de estirpes relacionadas. Um total de 58 estirpes de bactérias púrpuras não sulfurosas foram isoladas a partir de 108 amostras de fezes de galinhas na granja produtora de ovos do Departamento de Ciência Animal, Faculdade de Recursos Naturais da Universidade Prince of Songkla, Hat Yai, Tailândia. Depois de cultivar as bactérias em um substrato de glutamato/malato (GM), sem ácido glutâmico adicionado, sob luz (3000 lux) a 35 ºC durante 5 dias, os grânulos de biopolímeros intracelulares das bactérias foram observados utilizando um microscópio confocal (do inglês Confocal Laser Scanning Microscope - CLSM) com comprimentos de onda de excitação e emissão de 530 e 605 nm, respectivamente. A cromatografia gasosa (do inglês Gas chromatography - GC) foi realizada para uma análise quantitativa de PHB. Havia 5 estirpes, CH12, CH52, CH72, CH90 e CH92, que mostraram grânulos biopoliméricos quando submetidos ao CLSM, e PHB-5 acumulado de 1.7, 1.5, 1.4 and 1.8% (w w-1) do peso celular seco (do inglês cell dry weight - CDW), respectivamente. A análise da sequência do rDNA 16S da estirpe CH12 demonstrou uma alta correlação de homologia de 100% para aquela da estirpe NCIB8288 da Rhodopseudomonas palustris. Em relação às características taxonômicas e da análise da sequência do rDNA 16S, a estirpe CH12 foi identificada como Rps. palustris NCIB8288. A atividade da PHA sintase do extrato bruto da estirpe CH12 foi de 25 unidades/mL. As regiões conservadas puderam ser alinhadas e selecionadas entre 5 estirpes de Rhodopseudomonas palustris (BisA53, TIE-1, CGA009, HaA2 e BisB18). O produto purificado da reação em cadeia da polimerase - PCR foi obtido para estudos futuros.

Verification of conserved domain peptides derived from human antimicrobial peptide hGlyrichin / 军事医学

Objective To verify the antibacterial activity of the conserved domain derived from the novel human antimicrobial peptide-hGlyrichin .Methods Bioinformative analysis was performed and two peptides derived from hGlyrichin were synthesized which contained the conserved domain .Results Analysis of antimicrobial activities showed that these two peptides exhibited strong antibacterial activity which was inversely proportional to the length of the peptide within an eligible range.Despite the effective inhibition and killing of bacteria , the synthetic peptide segments had no hemolytic effect on human red blood cells .Conclusion These results indicate that a conserved domain exists in hGlyrichin , and that the peptides which contain this domain have strong antibacterial activity but are not toxic to human somatic cells .

Laparoscopic diaphragmatic hernioplasty in a dog / Hernioplastia diafragmática laparoscópica em cão

A diaphragmatic hernia is characterized by the passage of the abdominal viscera into the thoracic cavity, which may be congenital or acquired. Its treatment is achieved by surgical correction. When there is no tissue or in cases of herniation with a chronic disease, the use biological or synthetic implants is recommended. The objective of this study was to report a technique of laparoscopic diaphragmatic hernia repair using bovine pericardium preserved in a canine, using three portal accesses. Due to the large diaphragmatic defect, reduction with the aid of a network of preserved bovine pericardium in formaldehyde 4% was chosen. The mesh was sutured to the transversus abdominus muscle in two layers. The first layer was sutured using simple continuous pattern, and the second one using simple interrupted sutures. The patient collapsed and died 24 hours postoperatively. However, the purposed technique was feasible...

A hérnia diafragmática é caracterizada pela passagem das vísceras abdominais para a cavidade torácica, podendo ser de origem congênita ou adquirida, que exige o tratamento cirúrgico. Quando houver ausência de tecido ou em casos de herniação com evolução crônica, recomenda-se a utilização de implantes biológicos ou sintéticos. O objetivo deste trabalho é relatar a técnica de herniorrafia diafragmática laparoscópica com o uso de pericárdio bovino conservado em um canino, a partir do acesso laparoscópico com três portais. Devido ao grande defeito diafragmático, optou-se pela sua redução com o auxílio de implante de pericárdio bovino conservado em formaldeído a 4%, este fixado a musculatura diafragmática com sutura intracorpórea, utilizando para isso duas camadas de sutura ambas com náilon 0, a primeira contínua simples seguida de pontos isolados simples, em toda extensão da membrana conservada. Apesar do animal vir a óbito nas primeiras 24 horas do pós-operatório, a técnica adotada se mostrou viável...

Computational identification and systematic classification of novel GRAS genes in Isatis indigotica / 中国天然药物

Isatis indigotica Fort., belonging to Cruciferae, is one of the most commonly used plants in traditional Chinese medicine. The accumulation of the effective components of I. indigotica is related with its growth conditions. The GRAS genes are members of a multigene family of transcriptional regulators that play a crucial role in plant growth. Although the activities of many GRAS genes have long been recognized, only in recent years were some of them identified and functionally characterized in detail. In the present study, 41 GRAS genes were identified from I. indigotica through bioinformatics methods for the first time. They were classified into ten groups according to the classification of Arabidopsis and rice. The characterization, gene structure, conserved motifs, disordered N-terminal domains, and phylogenetic reconstruction of these GRASs were analyzed. Forty-three orthologous gene pairs were shared by I. indigotica and Arabidopsis, and interaction networks of these orthologous genes were constructed. Furthermore, gene expression patterns were investigated by analysis in methyl jasmonate (MeJA)-treated I. indigotica hairy roots based on RNA-seq data. In conclusion, this comprehensive analysis would provide rich resources for further studies of GRAS protein functions in this plant.

Composição nutricional de milho Bt ensilado com inoculante enzimobacteriano e avaliado em ovinos / Nutritional composition of GMO corn ensiled with bacterial inoculants and sheep digestibility

O objetivo foi avaliar as características agronômicas e químico-bromatológicas de dois híbridos de milho Bt (30F35H e CD397YH) ensilados com inoculante enzimobacteriano. Os teores de FDN foram semelhantes para todas as frações de planta, já o teor de FDA diferiu quanto à planta inteira e colmo, enquanto a lignina diferiu em relação à planta inteira, colmo e sabugo. A DIVMS não apresentou diferença entre os híbridos em nenhuma das frações. As silagens foram produzidas em silos experimentais (aproximadamente 200kg). As concentrações de MS, EE, FDN, NDT e DIVMS não mostraram diferença entre as silagens dos híbridos avaliados. Já os teores de MM, PB, FDA e lignina diferiram. Não houve diferença entre as silagens dos híbridos para os valores de NDT estimado e para a DIVMS. Também não foi observado efeito do inoculante sobre os valores de CHT, CNF, FDN e DIVMS. Conclui-se que o híbrido Pioneer foi superior ao Coodetec em produtividade de MV ha-1, porém a composição nutricional das silagens não diferiu na concentração de NDT e digestibilidade avaliadas em ovinos. Não houve efeito do uso de inoculante na digestibilidade da matéria seca e da fração fibra em detergente neutro das silagens.

The aim was to evaluate the agronomic characteristics and chemical composition of the two corn hybrids (30F35H and CD397YH) ensiled with enzymatic bacterial inoculants. NDF were similar for all plant fractions, since the ADF content differed as to the whole plant and stem, lignin differed in relation to the whole plant, stem and cob. IVDMD did not differ among treatments in any of the fractions. The silages were produced in experimental silos (approximately 200kg). The concentrations of MS, EE, NDF, IVDMD and TDN showed no difference between the silages of hybrids. Since the levels of MM, CP, ADF and lignin differed; there was no difference between hybrids for silage TDN and IVDMD. There was also no effect of the use of inoculants on the values of CHT, NFC, NDF and IVDMD. It is concluded that Pioneer was superior to Coodetec productivity of MV-1 ha. The nutritional composition of silages did not differ in the concentration of TDN and digestibility in sheep assessed. There was no effect of using inoculants on the digestibility of dry matter and neutral detergent fiber content of the silage.

Application of hexon gene sequencing of adenovirus in pathogen detection of epidemic keratoconjunctivitis / 中华实验眼科杂志

Background Epidemic keratoconjunctivitis is a common eye disease,and adenovirus is one of the common pathogens.The hexon protein,one main capsid protein of the virus,is an important target of antibody binding.Thus,sequencing the coding region of the hexon protein is an important way for adenovirus fast typing.Objective This study was to complete a molecular epidemiology survey of epidemic keratoconjunctivitis and investigate its association with adenovirus in Shanghai area by sequencing the coding region of hexon protein.Methods Two hundred and fourteen sacconjunctival swab specimens were collected from 214 patients with suspicious epidemic keratoconjunctivitis who visited Shanghai Eye Disease Prevention and Treatment Center and the clinical sites supervised by the Shanghai Prevention and Monitoring Office of Acute Hemerragic Conjunctivitis under the informed consent from January 2010 to December 2012.DNA was extracted from the specimens and then the 140 bp conserved sequence in hexon protein coding region was amplified by PCR initially to determine an adenovirus pathogen.Furtherly,956 bp conserved sequence of the hexon codind district was sequencied to clarify the serotype of adenovirus in the adenovirus-positive specimens.Results 50.93％ patients (109/214) were detected to be adenovirus-positive by generic PCR,in which AdV1 + was in 4 patiens,AdV2+ was in 33 patients,AdV3+ was in 15 patients,AdV4+ was in 12 patients,AdV8+ was in 19 patiens,AdV19+ was in 15 patients,AdV37+ was in 8 patients.The subgenus D adenoviruses,including AdV8+,AdV19+ and AdV37+ often resulted in corneal inflammation,pseudomembranous conjunctivitis and preauricular lymph nodes;while subgenus B adenovirus induced much frequent tract infection and less corneal response.Conclusions PCR-sequence of conserved region of hexon protein coding district is applicable for the detection and serotyping of adenovirus in epidemic keratoconjunctivitis.

Preparation and characterization of monoclonal antibody against hTSC29 pep-tides / 中国免疫学杂志

Objective:To prepare monoclonal antibody ( mAb ) against human testis-specific conserved gene ( hTSC29 ) peptides and characterize its immunological and biological features.Methods:According to bioinformatics analysis and prediction of the antigenicity, surface property, hydrophilicity and flexibility of hTSC29, a 18-amino acid residue partial peptide of hTSC29 was synthesized,then immunized the BALB/c mice for preparing antiserum.The mAb against hTSC29 was produced using the routine hybridoma technique.The properties of the mAb against hTSC29 were identified by ELISA, Western blot and immunohistochemistry staining.Results:After cell fusion and subcloning, one hybridoma cell lines secreting specific mAb against hTSC29 protein were obtaind.The Ig subclass of the mAb was IgG2b(κ).ELISA detection showed that the titer of mAbs in cultured was 1∶104.Western blot analysis proved that the mAb could specifically recognize Mr 60 000 protein in human testis total protein.The hTSC29 protein main located at circumference of spermatocyte and spermatid in human testis tissue by immunohistochemistry staining and immunofluorescence assay.Conclusion:One hybridoma cell lines which can secrete specific mAb against hTSC29 protein with high titers and specificity have been established successfully.The mAb will provide efficient tools for functional studies of hTSC29 expressed in spermatogenesis.