Molecular analysis of J-virus and Beilong virus using reverse genetics

The emergence of viruses in the family Paramyxoviridae, especially those such as Hendra virus and Nipah virus (NiV) that are zoonotic, highlighted the severity of disease that could be caused by infection with viruses belonging to this family. In addition to causing disease outbreaks, several newly discovered paramyxoviruses were found to have unique genetic features, which provoked renewed interest in the study of previously unclassified or uncharacterised viruses in this family. J-virus (JPV) was isolated from wild mice, in Queensland, Australia, in 1972, and has been suggested to be a natural respiratory pathogen of mice. Beilong virus (BeiPV), another paramyxovirus, was first isolated from human mesangial cells in Beijing, China, in 2003, and was subsequently detected in rat mesangial cells. Following initial characterisation, the genomes of JPV and BeiPV were found to contain two genes, SH and TM, not common to other paramyxoviruses, as well as an extended attachment protein gene. BeiPV has the largest genome in the family Paramyxoviridae, which is, in fact, larger than that of any other virus within the order Mononegavirales. The genetic material of paramyxoviruses is not amenable to manipulation via classical genetics; a reverse genetics approach was therefore employed to study the evolution and classification of JPV and BeiPV. Minireplicon systems utilising green fluorescent protein as a reporter were established for JPV, BeiPV and NiV, and were used to better assess the taxonomic status of JPV and BeiPV, and to determine the relationship between these viruses and henipaviruses, which also have exceptionally large genomes. These studies indicate that JPV and BeiPV are closely related and should be classified in the same genus and their replication and transcription machinery is different from that of the henipaviruses.To gain an understanding of the biology of JPV and BeiPV, viral surface proteins from JPV were expressed and evaluated. Chimeric JPV virions containing recombinant surface proteins were generated and electron microscopy was used to determine the localisation of the proteins encoded by those JPV genes which are uncommon in other paramyxoviruses. Analysis of the attachment protein gene of JPV indicated that the virus was able to assemble an exceptionally large protein (156 kDa) into the virion structure, providing evidence in support of the hypothesis that JPV and BeiPV may represent an ancient lineage of viruses within the family Paramyxoviridae. In order to determine tissue tropism of JPV during experimental infection and to aid future work with a full-length JPV infectious clone, a real-time PCR assay for JPV was developed and assessed on tissues collected from mice infected with JPV. A multiplex microsphere assay for JPV and BeiPV was developed and used to analyse the seroprevalence of these viruses in Australian and Malaysian rodents. Although there is currently no evidence for disease caused by JPV or BeiPV, this does not preclude the emergence of a zoonotic rodent paramyxovirus related to these viruses. If this were to occur, the tools for virus detection and serological monitoring are now established

Yongjia Xuanjue (665-713): An Introduction and Analysis of the Yongjia Anthology’s Method of Contemplative Practice

本文考察唐代著名僧人永嘉玄觉（665-713）的出家修道、探究佛法的历程及其历史与宗教背景，以《永嘉集》为中心，分析其修心方法及其思想来源。 永嘉玄觉虽然是中国中古时代著名的高僧之一，可是，他的生平以及归于他名下的两项重要著作，《永嘉集》与《证道歌》，在学术上，却并未得到应有的关注，相关研究并不多。他的思想一般被纳入正统的禅宗体系内加以叙述，且往往只占很少的分量。这些叙述与分析都强调玄觉受到禅宗六祖惠能（638-713）的启发而“觉悟”，重视长期以来被认为属于玄觉作品的《证道歌》，认为它运用高超的技巧，很好地阐发了禅宗的思想。而《永嘉集》的价值和意义则完全被遮蔽，一般只是把它看作为玄觉在“悟...This thesis explores the biography of a Tang era Buddhist monk named Yongjia Xuanjue 永嘉玄觉 (665-713), the historical and religious context of his monastic career, and his doctrinal background and approach to contemplative practice contained in his written work, the Yongjia Anthology 永嘉集. Although he is an important monastic figure of medieval Chinese Buddhism, Xuanjue’s biographies and the two w...学位：历史学硕士院系专业：人文学院_中国史学号：1032014115460

1861 - Kentucky Civil War Neutrality Proclamation

Document citation: Magoffin, B. Proclamation by the Governor of Kentucky, May 20, 1861 retrieved from Duke University Digital Repository https://idn.duke.edu/ark:/87924/r41g0m071.https://scholarworks.uni.edu/nhomefront/1005/thumbnail.jp

The Effect of Whole Body Vibration on Exercise-Induced Muscle Damage and Delayed-Onset Muscle Soreness

Current scientific evidence suggests that when whole body vibration (WBV) is used as a warm-up prior to performing eccentric exercise, delayed-onset muscle soreness (DOMS) is mitigated and strength loss recovers faster. These benefits were observed primarily in nonresistance-trained individuals. The aim of this study was to determine if WBV could mitigate soreness and expedite strength recovery for resistance-trained individuals when used as a warm-up prior to eccentric exercise. Thirty resistance-trained males completed 300 maximal eccentric contractions of the quadriceps after warming up with (WBV) or without (CON) WBV. Both CON and WBV experienced significant isometric (27.8% and 30.5%, respectively) and dynamic (52.2% and 47.1%, respectively) strength loss immediately postexercise. Isometric strength was significantly depressed after 24 hours in the CON group (9.36% p \u3c 0.01), but not in the WBV group (5.8% p = 0.1). Isometric strength was significantly depressed after 48 hours in the CON group (7.18% p \u3c 0.05), but not in the WBV group (4.02% p = 0.25). Dynamic strength was significantly decreased in both the CON and WBV groups both at 24 hours (19.1% p \u3c 0.001, and 16.1% p \u3c 0.001, respectively), 48 hours (18.5% p \u3c 0.01, and 14.5% p \u3c 0.03), and 1 week postexercise (9.3% p = 0.03, and 3.5%, respectively). Pain as measured by visual analog scale (VAS) was significant in both CON and WBV groups at 24 and 48 hours postexercise, but the WBV experienced significantly less soreness than the CON group after 24 hours (28 mm vs. 46 mm p \u3c 0.01 respectively), and 48 hours (38 mm vs. 50 mm p \u3c 0.01). Pain as measured by pain pressure threshold (PPT) increased significantly in both groups after 24 and 48 hours, but there was no difference in severity of perceived soreness. The use of WBV as a warm-up may mitigate DOMS but does not appear to expedite the recovery of strength in the days following eccentric exercise in resistance-trained individuals

Cholestasis and meconium ileus in infants with cystic fibrosis and their clinical outcomes

Objective To identify the incidence and outcomes of cholestasis and meconium ileus (MI) in infants with cystic fibrosis (CF). Design Retrospective cohort study. Setting Single-centre study. Patients From January 1986 to December 2011, 401 infants with CF (69 with MI) presented to our centre. Main outcome measurements (1) incidence of cholestasis, (2) identification of risk factors for cholestasis, (3) association between the presence of cholestasis and MI and the development of clinically significant CF-associated liver disease (CFLD) defined as multilobular cirrhosis with portal hypertension. Results Cholestasis occurred in 23 of 401 infants (5.7%). There was a significantly higher incidence of cholestasis in infants with MI (27.1%) compared to those without MI (1.2%) (p Conclusions Cholestasis is an uncommon condition in CF affecting only 5.7% of the screened newborn CF population. The greatest risk factor for developing cholestasis is the presence of MI. However, the presence of MI appears not to be associated with the development of CFLD. An effect of neonatal cholestasis on the development of CFLD cannot be excluded by this study

A functional link between bone morphogenetic proteins and insulin-like peptide 3 signaling in modulating ovarian androgen production

Bone morphogenetic proteins (BMP) are firmly implicated as intra-ovarian regulators of follicle development and steroidogenesis. Here we report a microarray analysis showing that treatment of cultured bovine theca cells (TC) with BMP6 significantly (>2-fold; P<0.01) up- or down-regulated expression of 445 genes. Insulin-like peptide 3 (INSL3) was the most heavily down-regulated gene (-43-fold) with CYP17A1 and other key transcripts involved in TC steroidogenesis including LHCGR, INHA, STAR, CYP11A1 and HSD3B1 also down-regulated. BMP6 also reduced expression of NR5A1 encoding steroidogenic factor-1 known to target the promoter regions of the aforementioned genes. Real-time PCR confirmed these findings and also revealed a marked reduction in expression of INSL3 receptor (RXFP2). Secretion of INSL3 protein and androstenedione were also suppressed suggesting a functional link between BMP and INSL3 pathways in controlling androgen synthesis. RNAi-mediated knockdown of INSL3 reduced INSL3 mRNA and secreted protein level (75 and 94%, respectively) and elicited a 77% reduction in CYP17A1 mRNA level and 83% reduction in androstenedione secretion. Knockdown of RXFP2 also reduced CYP17A1 mRNA level (81%) and androstenedione secretion (88%). Conversely, treatment with exogenous (human) INSL3 increased androstenedione secretion ~2-fold. The CYP17 inhibitor abiraterone abolished androgen secretion and reduced expression of both INSL3 and RXFP2. Collectively, these findings indicate a positive autoregulatory role for INSL3 signaling in maintaining thecal androgen production, and visa versa. Moreover, BMP6-induced suppression of thecal androgen synthesis may be mediated, at least in part, by reduced INSL3-RXFP2 signaling

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Thecal cell sensitivity to luteinizing hormone and insulin in polycystic ovarian syndrome

This study examined whether a defect of steroid synthesis in ovarian theca cells may lead to the development of PCOS, through contributions to excess androgen secretion. Polycystic ovarian syndrome (PCOS) is one of the leading causes of infertility worldwide affecting around 1 in 10 of women of a reproductive age. One of the fundamental abnormalities in this syndrome is the presence of hormonal irregularities, including hyperandrogenemia, hyperinsulinemia and hypersecretion of luteinizing hormone (LH). Studies suggest that insulin treatment increases progesterone and androstenedione secretion in PCOS theca cells when compared to insulin treated normal theca cells. Furthermore the augmented effects of LH and insulin have been seen to increase ovarian androgen synthesis in non-PCOS theca cultures whilst also increasing the expression of steroidogenic enzymes specific to the PI3-K pathway. Our examination of primary thecal cultures showed an increase in both the expression of the steroidogenic enzyme CYP17 and androgen secretion in PCOS theca cells under basal conditions, when compared to non-PCOS cells. This was increased significantly under treatments of LH and insulin combined. Our results support the previous reported hypothesis that a dysfunction may exist within the PI3-K pathway. Specifically, that sensitivity exists to physiological symptoms including hyperinsulinemia and hyper secretion of LH found in PCOS through co-stimulation. The impact of these findings may allow the development of a therapeutic target in PCOS