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The etiology of systemic lupus erythematosus (SLE) involves a complex interaction of genetic and environmental factors. Investigations have shown that environmentally driven epigenetic changes contribute to the etiology of SLE. Here, we hypothesize that aberrant DNA methylation may contribute to the activation of the immune machinery and trigger lupus disease activity. A whole genome methylation array was applied to investigate the DNA methylation changes between 12 pairs of active SLE patients and healthy controls. The results were further confirmed in 66 SLE patients, 102 healthy controls. The methylation statuses of the IL10 and IL1R2 genes were significantly reduced in the SLE patient samples relative to the healthy controls (age-adjusted odds ratios, 64.2 and 16.9, respectively, P<0.0001). There was a trend toward SLE patients having hypomethylated IL10 and IL1R2 genes accompanied by greater disease activity. We observed that the methylation degree of IL10 and IL1R2 genes were reduced in the rheumatoid arthritis (RA) patients as well but the hypomethylation change was more significant in IL1R2 genes than in the IL10 genes in RA patients. This study demonstrated that DNA hypomethylation might be associated with SLE. Hypomethylated IL10 and IL1R2 genes may provide potential epigenetic markers as clinical predictors for autoimmune diseases. [ABSTRACT FROM AUTHOR]

Background: Despite earlier studies demonstrating in vitro propagation of solid tumour cancer stem cells (CSCs) as non-adherent tumour spheres, it remains controversial as to whether CSCs can be maintained in vitro. Additional validation of the CSC properties of tumour spheres would support their use as CSC models and provide an opportunity to discover additional CSC cell surface markers to aid in CSC detection and potential elimination.Methods: Primary tumour cells isolated from 13 surgically resected colon tumour specimens were propagated using serum-free CSC-selective conditions. The CSC properties of long-term cultured tumour spheres were established and mass spectrometry-based proteomics performed.Results: Freshly isolated CD133(+) colorectal cancer cells gave rise to long-term tumour sphere (or spheroids) cultures maintaining CD133 expression. These spheroid cells were able to self-renew and differentiate into adherent epithelial lineages and recapitulate the phenotype of the original tumour. Relative to their differentiated progeny, tumour spheroid cells were more resistant to the chemotherapeutic irinotecan. Finally, CD44, CD166, CD29, CEACAM5, cadherin 17, and biglycan were identified by mass spectrometry to be enriched in CD133(+) tumour spheroid cells.Conclusion: Our data suggest that ex vivo-expanded colon CSCs isolated from clinical specimens can be maintained in culture enabling the identification of CSC cell surface-associated proteins. [ABSTRACT FROM AUTHOR]

A novel method for generating an antigen-specific cancer vaccine and immunotherapy has emerged using a DNA vaccine. However, antigen-presenting cells (APCs) have a limited life span, which hinders their long-term ability to prime antigen-specific T cells. Connective tissue growth factor (CTGF) has a role in cell survival. This study explored the intradermal administration of DNA encoding CTGF with a model tumor antigen, human papilloma virus type 16 E7. Mice vaccinated with CTGF/E7 DNA exhibited a dramatic increase in E7-specific CD4+ and CD8+ T-cell precursors. They also showed an impressive antitumor effect against E7-expressing tumors compared with mice vaccinated with the wild-type E7 DNA. The delivery of DNA encoding CTGF and E7 or CTGF alone could prolong the survival of transduced dendritic cells (DCs) in vivo. In addition, CTGF/E7-transduced DCs could enhance a higher number of E7-specific CD8+ T cells than E7-transduced DCs. By prolonging the survival of APCs, DNA vaccine encoding CTGF linked to a tumor antigen represents an innovative approach to enhance DNA vaccine potency and holds promise for cancer prophylaxis and immunotherapy.Gene Therapy (2008) 15, 1007–1016; doi:10.1038/gt.2008.25; published online 20 March 2008 [ABSTRACT FROM AUTHOR]

Alphavirus vectors have emerged as a promising strategy for the development of cancer vaccines and gene therapy applications. In this study, we used the replication-defective vaccine vector SIN replicon particles from a new packaging cell line (PCL) to develop SIN replicon particles encoding calreticulin (CRT) linked to a model tumor antigen, human papillomavirus type 16 (HPV16) E7 protein. The linkage of CRT to E7 in SIN replicon particles resulted in a significant increase in E7-specific CD8+ T-cell precursors and a strong antitumor effect against E7-expressing tumors in vaccinated mice. SINrep5-CRT/E7 replicon particles enhanced presentation of E7 through the major histocompatibility complex (MHC) class I pathway by infecting dendritic cells (DCs) directly and pulsing DCs with lysates of cells infected by SINrep5-CRT/E7 replicons. Vaccination of immunocompromised (BALB/c nu/nu) mice with SINrep5-CRT/E7 replicon particles also generated significant reduction of lung tumor nodules, suggesting that antiangiogenesis may contribute to the antitumor effect of SINrep5-CRT/E7 replicon particles. Furthermore, SINrep5-CRT/E7 replicon particles generated long-term in vivo tumor protection effects and antigen-specific memory immunities. We concluded that the CRT strategy used in the context of SIN replicon particles facilitated the generation of a highly effective vaccine for cancer prophylaxis and immunotherapy.Cancer Gene Therapy (2006) 13, 873–885. doi:10.1038/sj.cgt.7700956; published online 28 April 2006 [ABSTRACT FROM AUTHOR]

The feeding ecology of leptostracans is not well-known. In the present study, species of Nebalia (Crustacea: Leptostraca) were captured every month for 1 year from Lobster Bay, Hong Kong, using baited traps. Captured individuals fell into a size range of 0.36–3.64 mm and 0.95–5.52 mm in carapace and total lengths, respectively. They were divided into different reproductive stages based on the morphology of the second pair of antennae and thoracopods. Sexual dimorphism becomes apparent between 0.7–1.5 mm (CL) and 1.4–3.2 mm (TL). Most individuals collected were sexually differentiated, but immature (79.7% of the captured individuals), followed by post-ovigerous (11.4%) and ovigerous (8.9%) females. Sexually undifferentiated juveniles were rare (3.9%). The sex ratio of the immature individuals approached 1:1 with a propensity towards male dominance. Not one mature male was captured. Although females with setose thoracopod endopods, indicating that they were ovigerous, were captured frequently, only two were carrying eggs. The two brooding females had clutch sizes of 11 and 17 eggs. Alternate peaks in relative abundance of ovigerous and post-ovigerous females were identified, the latter following the former. Reproductive activities occurred mainly in April–May and October–December, indicating that two cohorts are recruited. Catches using baited traps could be explained by the reproductive season of Nebalia sp. High catches were usually associated with the months of highest reproductive activity, low catches with peaks in post-ovigerous females. It is therefore concluded that not all individuals at various reproductive stages are attracted to carrion and hence adopt a scavenging feeding mode. Catches of Nebalia sp. in Hong Kong possibly reflect their annual cycle of reproductive activity [ABSTRACT FROM AUTHOR]

Background: To evaluate the influence of hypoxia and molecular events in endothelial and embryonic stem cells. Materials and Methods: Human umbilical vein endothelial cells (HUVECs) and mouse embryoid body (EB) cells were subjected to hypoxic conditions for different time courses. DNA fragmentation assay, quantification of apoptotic cells by TUNEL assay measured by flowcytometry, and Western blot analysis for the molecular events of apoptosis were performed. Results: DNA fragmentation could be identified under hypoxic conditions in HUVECs and mouse EBs. The DNA fragmentation increased when the hypoxic interval was extended. In situ internucleosomal DNA fragmentation-TUNEL assay also found that the percentages of apoptotic cells increased gradually in HUVECs and mouse EBs when the hypoxic interval was extended. Furthermore, the levels of expression of p53 and Bax both increased in hypoxic conditions. Conclusions: Hypoxia increases both HUVEC and mouse EB apoptosis, which is associated with increase in p53/Bax expression. [ABSTRACT FROM AUTHOR]

Study design:A case report.Objectives:This study discusses a case of spinal segmental myoclonus caused by thoracic myelopathy, mimicking hiccup spasms. Spinal myoclonus caused by thoracic myelopathy is extremely rare. It can be misdiagnosed as chronic intractable hiccups due to similar clinical manifestations.Setting:Korea University Anam Hospital, Seoul, Republic of Korea.Methods:A 42-year-old man presented with a history of involuntary jerky movement of the upper abdominal wall muscles that had been continuing for over 3 years. A neurological examination, brain computed tomography and electroencephalogram did not reveal a cause of the symptoms. Electromyography was performed on the abdominal muscles and the findings revealed were compatible with spinal myoclonus. The spinal myoclonus had started in the abdominal muscles, with a spinal magnetic resonance imaging revealing a disc protrusion compressing the anterior spinal cord.Results:The cause of the spinal myoclonus was determined to be spinal myelopathy due to mild T7 disc protrusion. The patient refused surgical or invasive interventions and was conservatively treated with clonazepam. The symptoms were reported to be less frequent following the treatment.Conclusion:Compressive myelopathy developed from disc protrusion may cause spinal myoclonus mimicking as hiccup spasms. [ABSTRACT FROM AUTHOR]