Identification and characterization of the duck enteritis virus (DEV) US2 gene.

The US2 protein has been reported to contribute to duck enteritis virus (DEV) infection; however, its kinetics and localization during infection, and whether it is a component of virion, have not been previously reported. To elucidate the function of DEV US2, US2 was amplified by polymerase chain reaction (PCR) and inserted into pET-32a(+); this was expressed, the recombinant US2 protein was purified, and a polyclonal antibody generated. In addition, the kinetics and localization of the US2 gene and protein were determined by quantitative real-time fluorescent PCR, ganciclovir (GCV), and cycloheximide (CHX) treatment, western-blot, and indirect immunofluorescence assay. The packaging of US2 into DEV virions was revealed by a protease protection assay. US2 was found to be transcribed 24 h post-infection (pi) and peaked at 72 h pi; the US2 protein was detected 48 h pi, except in the presence of GCV or CHX. US2 was packed into virions and also localized to the plasma membrane and cytoplasm in infected cells. The results showed that the DEV US2 is a late gene, and that its encoding protein could be a tegument component localized mainly in the cytoplasm. This study provides useful data for the further analysis of DEV US2, including an explanation for the genetic conservation among alphaherpesviruses.

miR-96 inhibits cardiac hypertrophy by targeting growth factor receptor-bound 2.

Increasing evidence has indicated that microRNAs are involved in the pathogenesis of cardiac hypertrophy. However, whether miR-96 is involved in heart diseases, particularly cardiac hypertrophy, remains unclear. In this study, we found that miR-96 is a negative regulator of cardiac hypertrophy. In primary cardiomyocytes, overexpression of miR-96 inhibited phenylephrine-induced cardiomyocyte hypertrophy and decreased the mRNA expression of cardiac hypertrophy markers such as atrial natriuretic factor and ß-myosin heavy chain. Interestingly, we found that growth factor receptor-bound 2 is a direct target of miR-96, which is a negative regulator of cardiac hypertrophy. Overexpression of miR-96 in cardiomyocytes led to reduced growth factor receptor-bound 2 expression. More importantly, miR-96 repressed the extracellular-regulated protein kinase signaling pathway by targeting growth factor receptor-bound 2 in cardiomyocytes. Our data demonstrate that miR-96 is a negative regulator of cardiac hypertrophy and extracellular-regulated protein kinase signaling, thus offering a new therapeutic strategy for cardiac hypertrophy.

Characterization of Riemerella anatipestifer CH-1 gldJ gene and GldJ protein.

Riemerella anatipestifer (RA) CH-1, a highly virulent field strain, was isolated and identified by our laboratory. The gldJ gene was conserved in RA, and it had a typical TATA promoter region and AU-rich sequence within the 5' untranslated region. The GldJ protein was an outer-membrane lipoprotein with a signal peptide cleavage site between amino acids 20 and 21. GldJ was also a member of proteins involved in gliding motility. The RA GldJ protein had 16 phosphorylation sites and 4 N-glycosylation sites. These functional sites played an important role in the posttranslational modification of the GldJ protein. In addition, the GldJ protein of RA CH-1 had strong immunogenicity. Fifteen B-cell epitopes were identified in the GldJ protein, which might be a good biological material for the development of a subunit vaccine and diagnostic reagents. The GldJ protein also had the activity of formylglycine-generating sulfatase enzyme. The 3-dimensional structure models of GldJ were constructed based on 2 templates using the SwissModel automatic modeling mode in the SWISS-MODEL workplace. Here, we characterized the RA gldJ gene and GldJ protein to contribute to the functional annotation for the GldJ protein.

Bioinformatic analysis and characteristics of glycoprotein C encoded by the newly identified UL44 gene of duck plague virus.

Glycoprotein C is one of the duck plague virus (DPV) glycoproteins and is encoded by the DPV UL44 gene. DPV glycoprotein C (DPV-gC) comprises 431 amino acids with a putative molecular mass of 47.35 kDa. Sequence analysis indicated that the protein possesses typical characteristics of type-I membrane glycoproteins, containing an N-terminal signal sequence, an external domain, a C-terminal membrane anchor region, and a short cytoplasmic domain. Comparisons of 22 alphaherpesvirus-gC protein sequences revealed eight conservative Cys-residue sites, which may play a crucial role in the biological functions and structural stabilization of the DPV-gC protein. Estimates of potential antigenic epitopes and secondary structure identified four B cell dominant epitopes, which are located at amino acids 68-71, 87-91, 369-352, and 372-374. A model for the structure of DPV-gC was derived by associating its predicted secondary and three-dimensional structures. In conclusion, these results will provide a basis for further functional studies of DPV-gC, establishing novel clinical diagnoses of DPV, and in the development of a new DPV vaccine.

Extrapulmonary infections caused by a dominant strain of Mycobacterium massiliense (Mycobacterium abscessus subspecies bolletii).

A single strain of Mycobacterium massiliense (BRA 100), a subspecies of the Mycobacterium abscessus complex, has been responsible for an epidemic of post-surgical infections in Brazil. Outside Brazil, this is the first report to describe a single emerging strain of M. massiliense (TPE 101) associated with extrapulmonary infections. This phenomenon may be underestimated because sophisticated molecular typing of M. abscessus is not routinely performed. Our molecular epidemiology study was triggered by an outbreak investigation. Nine case isolates were grown from the surgical sites of nine mostly paediatric patients receiving operations from 2010 to 2011. All available non-duplicated isolates of M. abscessus during this period were obtained for comparison. Mycobacteria were characterized by multilocus sequence analysis (MLSA), repetitive sequence PCR (rep-PCR) and pulsed-field gel electrophoresis (PFGE). Of 58 isolates of M. abscessus overall, 56 were clinical isolates. MLSA identified 36 of the isolates as M. massiliense. All case isolates were indistinguishable by PFGE and named the TPE 101 pulsotype. Of the stored strains of M. abscessus, TPE 101 strains were over-represented among the control surgical wound (7/7, 100%) and subcutaneous tissue isolates (4/5, 80%) but rare among the respiratory isolates (1/16, 6%) and absent from external skin, ocular and environmental samples. In conclusion, a unique strain of M. massiliense has emerged as a distinctive pathogen causing soft tissue infections in Taiwan. Further study to identify whether this is due to an occult common source or to specific virulence factors dictating tissue tropism is warranted.

In vitro antioxidant and photo-oxidant properties of dipyridamole.

The in vitro antioxidant and photo-oxidant activity of dipyridamole was studied by its effect on superoxide- and singlet oxygen-mediated photohemolysis and viability of neutrophils. Dipyridamole was found to be phototoxic when examined by the photohemolysis on human erythrocytes and on linoleic acid as lipid peroxidation model at concentrations above 3.0 x 10(-5) M. On the contrary, when lower concentrations (1.0 x 10(-5) to 1.0 x 10(-6) M) were used, dipyridamole showed a protector action against singlet oxygen-mediated photohemolysis by other phototoxic compounds like triamterene. This antioxidant property is proposed to result from quenching of triamterene mediated by fluorescence energy transfer. Auto-oxidation and fluorescence-energy transfer is clearly an important mechanism for protection for this drug.

[Effect of ethanol adminstration in the consumption of proteins and nitrogen balance in normal subjects with normoprotein diet]. / Efecto de la administración de etanol en la utilización protéica y balance nitrogenado en sujetos normales con dietas normoprotéicas.

The effect of excessive alcohol intake on the protein requirements and metabolism in normal subjects has not been clearly determined. In this study we measured the nitrogen balance, the hematrocrit, the hemoglobin, the serum albumin, the cholesterol, and the plasmatic amino acids in 7 non-alcoholic subjects of 25 +/- 5 years of age. A comparison was made of a diet containing 0.8 g of protein per k of weight and 40 Kcal per k of weight administered during 11 days with a period of the same length in which the 1.400 Kcal provided during the control period by carbon hydrates was provided by ethanol (200 g). During the alcoholic period no importants changes were observed in the nitrogen balance, a tendency towards greater positivity being registered. There was a decrease in the serum albumin of 4, 69 +/- 0.31 vs 3, 90 +/- 0,32 g/100 ml and an increase in globulin 1,74 +/- 0,70 vs 2,69 +/- 0,22 g/100 ml. The results showed that in a short period of time the excessive alcohol intake in normal subjects does not increase the protein requirements in spite of a decrease in the serum albumin being observed.