Expression of arylalkylamine n-acetyltransferase (AANAT) and acetylserotonin o-methyltransferase (ASMT) in the corpus luteum of pregnant sows and synthesis of melatonin in luteal cells.

In vertebrates, melatonin is mainly synthesized from serotonin in the pineal gland. Many reports have documented that melatonin is also synthesized in the extra-pineal tissues, but the synthesis of melatonin in the corpus luteum (CL) of pregnant sows has never been studied. The objectives of this study were to evaluate the expression of melatonin-synthesizing enzymes, arylalkylamine N-acetyltransferase (AANAT) and acetylserotonin O-methyltransferase (ASMT), in the CL of sows during pregnancy and to investigate the synthesis of melatonin in luteal cells. Results showed that AANAT and ASMT were both expressed in the CL of sows during pregnancy, higher levels were observed in the early- and mid-stage CL, and the lowest abundance was found in the regressing CL (later-stage). The immunostaining for AANAT and ASMT was predominantly localized in the large luteal cells of porcine CL during pregnancy. Furthermore, melatonin was synthesized in luteal cells from serotonin in a dose- and time-dependent manner. And the expressions of AANAT and ASMT were upregulated by serotonin in luteal cells. In addition, progesterone (P4) secretion and cell viability were promoted in luteal cells treated with serotonin, and the stimulatory effects were blocked by luzindole (a non-selective MT1 and MT2 antagonist). Finally, the expressions of MT1 and MT2 were augmented by serotonin in luteal cells. In conclusion, this study demonstrates for the first time the developmental expression of AANAT and ASMT in the CL and a local synthesis of melatonin in luteal cells of pregnant sows, and suggests a paracrine and/or autocrine role for melatonin in luteal function.

Transmissible gastroenteritis virus nsp7 protein localized in the cytoplasm down-regulates interleukin 8 expression in porcine intestinal epithelial cell.

Transmissible gastroenteritis virus (TGEV) is an important pathogen in swine that is responsible for substantial economic losses. Previous studies suggest that the TGEV non-structural protein 7 (nsp7) plays an important role in the viral assembly process. However, the subcellular localization and other functions of the TGEV nsp7 protein are still unclear. In this study we have examined the subcellular localization and other functions of TGEV nsp7 protein through analysis of its effects on cell growth, cell cycle progression, interleukin 8 (IL-8) expression, and NF-κB activation. Our results showed that the nsp7 protein is localized in the cytoplasm and has no effect on intestinal epithelial cells (IECs) growth, cell cycle, and cyclin A expression. Further studies showed that TGEV nsp7 protein had no effect on GRP78 expression, could not induce endoplasmic reticulum (ER) stress and activate NF-κB activity. Interestingly, the IECs expressing nsp7 protein secreted lower levels of IL-8 than control cells. This is the first report to demonstrate the subcellular localization and novel functions of TGEV nsp7 protein. These findings provide novel information about the function of the poorly characterized TGEV non-structural protein 7.

Modified recombinant adenoviruses increase porcine circovirus 2 capsid protein expression and induce enhanced immune responses in mice.

Porcine circovirus type 2 (PCV2) is the primary viral pathogen of porcine circovirus associated disease (PCVAD) and vaccination is an important method to prevent and control the disease. The expression of PCV2 capsid protein (Cap) in adenovirus vector system has been investigated, but the poor immune responses limit its application. In this study, transcriptional enhancer element largest intron of the human cytomegalovirus (Intron A) and woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) were applied to increase the immunogenicity of PCV2 Cap adenovirus-based vaccine. Western blot and indirect immunofluorescence assay (IFA) analysis showed that modified adenoviruses with Intron A and WPRE alone or both could significantly increase the expression of Cap compared to the unmodified adenoviruses. Furthermore, the humoral and cellular immune responses of the constructed recombinant adenoviruses were evaluated in mice. Indirect ELISA, virus neutralizing test and western blot showed that modified adenoviruses elicited higher humoral immune responses than unmodified adenovirus, and Intron A-WPRE-modified virus immunized group had better immune response than the others. Besides, the results of lymphocyte proliferation response and cytokines release assay showed that enhanced cellular immune responses were induced by modified adenoviruses. These results demonstrated that Intron A and WPRE significantly improved the expression of the Cap protein in adenovirus vector system and enhanced the immune responses in mice, making the adenovirus vector system more applicable against PCV2.

Porcine epidemic diarrhea virus M protein blocks cell cycle progression at S-phase and its subcellular localization in the porcine intestinal epithelial cells.

Porcine epidemic diarrhea (PED) caused by virulent strains of porcine epidemic diarrhea virus (PEDV) is a highly contagious enteric disease of swine characterized by severe enteritis, vomiting, and watery diarrhea. This study investigates the subcellular localization and function of PEDV M protein through examination of its effects on cell growth, cell cycle progression, and interleukin 8 (IL-8) expression. Our results revealed that the PEDV M protein is localized throughout the cytoplasm. The M protein altered swine intestinal epithelial cell line (IEC) growth and induced cell cycle arrest at the S-phase via the cyclin A pathway. The S-phase arrest is associated with a decrease in level of cyclin A. Furthermore, our results revealed that the M protein of PEDV does not induce endoplasmic reticulum (ER) stress and does not activate NF-κB which is responsible for IL-8 and Bcl-2 expression. This is the first report to demonstrate that the PEDV M protein is localized in the whole cell and induces cell cycle arrest at the S-phase. This study provides novel findings in the function of M proteins of PEDV.

Immunogenicity of the envelope GP3 protein of porcine reproductive and respiratory syndrome virus displayed on baculovirus.

Porcine reproductive and respiratory syndrome virus (PRRSV) has been recognized as one of the most important pathogens of pigs throughout the world. The minor envelope protein GP3 of PRRSV plays an important role in clearing of the virus infection and protecting the animals. In this study, a recombinant baculovirus (BacSC-GP3) expressing His6-tagged GP3 with the transmembrane (TM) and cytoplasmic (CT) domains of envelope protein gp64 was constructed and its immunogenicity was evaluated in mouse and piglet models. The His6-tagged GP3 was successfully displayed on the surface of virions as well as virus-infected Sf-9 cells. The animals immunized with BacSC-GP3 gave a slightly higher (piglets) up to a markedly higher (mice) humoral and lymphocyte proliferation responses than those that received a commercial killed vaccine. This is the first study on the immunogenicity of recombinant GP3-baculovirus, which indicates that the latter can represent an alternative strategy for developing a more effective PRRSV vaccine.

Association of the PTPN22 gene (-1123G > C) polymorphism with rheumatoid arthritis in Chinese patients.

This study aimed at examining the association of the single nucleotide polymorphism (SNP) in the protein tyrosine phosphatase gene (PTPN22) with the risk of rheumatoid arthritis (RA) in a Chinese population. A total of 200 RA patients and age and gender-matched healthy controls were recruited. Their genotypes and allelic frequency were determined by the TaqMan-MGB probe-based polymerase chain reaction (PCR). The frequencies of the CC genotype and C allele in RA patient group were significantly higher than that of controls (P < 0.01 or P < 0.05) with an odds ratio of 1.67, respectively. These data suggest, the CC genotype and C allele of the -1123G > C in the PTPN22 gene are associated with an increased risk for RA in Chinese population. Therefore, the CC genotype and C allele of the -1123G > C in the PTPN22 gene may be used as a genetic marker for the predisposition of RA in Chinese.

Conducting a factory or plant visit.

Plant visits are an invaluable tool in the management of work-related skin disorders, as they are the means by which a dermatologist gains firsthand information about the circumstances under which a work-related skin disorder has arisen in a plant or factory. In order to manage the workplace hazards which are responsible for skin disease, these must be identified and evaluated, and recommendations made about their control. While it may be possible to do this vicariously by questioning, examining, and investigating affected workers in the office, site visits to the plant or factory are sometimes necessary for the optimal assessment and management of such cases, as they often provide valuable information and insights not forthcoming from the history, examination, and investigations. This article explores the rationale behind plant visits, and discusses the pre-visit assessment, pre-visit preparation, the plant visit itself, and post-visit responsibilities. Practical advice is given for conducting each of these stages.

Beta-1,3-D-glucan gel in the treatment of solar keratoses.

beta-1,3-D-glucans are yeast-derived carbohydrate polymers which have been shown to be potent immunoresponse modulators which promote the regression of certain tumours. To date there is no published data concerning the efficacy of topical beta-1,3-D-glucan in the treatment of solar keratoses. This randomized double-blind prospective pilot study of 20 patients was performed to investigate the efficacy and skin tolerance of beta-1,3-D-glucan gel versus placebo in the treatment of solar keratoses. The results of this study showed no significant benefit in using beta-1,3-D-glucan gel over placebo in reducing counts of solar keratoses. No adverse effects were reported by any patient at any stage of the trial.

Pretibial myxoedema presenting as a scar infiltrate.

A 41-year-old man developed pretibial myxoedema localized to scars following treatment for hyperthyroidism. Pre-existing scars on both lower legs, present for more than 20 years, were infiltrated with firm and nonpitting nodules and plaques over his shins. The clinical presentation of this patient highlights pretibial myxoedema as a cause of scar infiltration. The pathogenesis of pretibial myxoedema is reviewed.

Widespread plaque psoriasis responsive to mycophenolate mofetil.

A woman with a long history of widespread plaque psoriasis unresponsive to and/or intolerant of phototherapy, retinoids, methotrexate and cyclosporin was successfully treated with mycophenolate mofetil. Remission was maintained on doses between 1 and 1.5 g/day for 18 months without adverse effects.