Lipoteichoic acid enhances IL-6 production in human synovial fibroblasts via TLR2 receptor, PKCδ and c-Src dependent pathways

Patients with rheumatoid arthritis (RA) are at increased risk of developing infections and appear to be particularly susceptible to septic arthritis. Lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria is an amphiphilic, negatively charged glycolipid. However, the effects of LTA on human synovial fibroblasts are largely unknown. We investigated the signaling pathway involved in IL-6 production stimulated by LTA in rheumatoid arthritis synovial fibroblasts (RASF). LTA caused concentration- and time-dependent increases in IL-6 production. LTA-mediated IL-6 production was attenuated by Toll-like receptor 2 (TLR2) monoclonal antibody or siRNA. Pretreatment with PKCδ inhibitor (rottlerin), c-Src inhibitor (PP2), AP-1 inhibitor (tanshinone IIA) and NF-κB inhibitor (PDTC and TPCK) also inhibited the potentiating action of LTA. However, focal adhesion kinase (FAK) mutant and siRNA did not affect LTA-mediated IL-6 production. Stimulation of cells with LTA increased the PKCδ and c-Src phosphorylation and kinase activity. LTA increased the accumulation of p-c-Jun and p-p65 in the nucleus, as well as AP-1 and NF-κB luciferase activity. LTA-mediated increase of AP-1 and NF-κB luciferase activity was inhibited by rottlerin and PP2 or TLR2 and PKCδ siRNA or c-Src mutant. Our results suggest that LTA-increased IL-6 production in human synovial fibroblasts via the TLR2 receptor, PKCδ, c-Src, AP-1 and NF-κB signaling pathways.     

ONCOLYTIC HERPES SIMPLEX VIRUS 1 (HSV-1) VECTORS: INCREASING TREATMENT EFFICACY AND RANGE THROUGH STRATEGIC VIRUS DESIGN

Viruses have long been considered potential anticancer treatments. Wild-type viruses have been tested as anticancer agents in clinical trials since the 1960s. The possibility of viral oncolysis as an alternate cancer therapy was transformed by the emergence of modern genetic engineering. The herpes simplex virus (HSV) family offers particular advantages for use as a viral oncolytic. The engineered vectors that make up oncolytic HSVs (oHSVs) have demonstrated remarkable safety in clinical trials, with some evidence of efficacy. The past decade has seen a focus on increasing the efficacy of oncolytic vectors by adding exogenous transgenes to enhance tumor destruction. The current paper describes the various strategies for engineering HSV for increased cancer tissue specificity and efficacy. Presented are the rationale, preclinical data and clinical data where available. This is meant to illustrate a basic framework for the development of a novel therapy meant to exploit the viral life cycle for the killing of cancer.     

Evaluation of internal reliability in the presence of inconsistent responses

Background  

We aimed to assess the impact of inconsistent responses on the internal reliability of a multi-item scale by developing a procedure to adjust Cronbach's alpha.     

Understanding Employers’ Hiring Intentions in Relation to Qualified Workers with Disabilities: Preliminary Findings

Introduction As part of the planning process for a larger survey study to examine factors affecting employers’ intention to hire and hiring of people with disabilities, a series of three semi-structured focus groups were held with key hiring decision makers, such as Human Resources directors, Chief Operating Officers (COOs), or Chief Executive Officers (CEOs) of small, medium, and large Seattle area companies. Aim The chief goals of the focus groups were to elicit and refine the participants’ beliefs, normative influences, and perceived control relative to hiring workers with disabilities. Method Narrative data obtained from the focus group discussion were examined using the Theory of Planned Behavior (TPB) to identify themes expressed by the focus group participants within the context of company size. Results Themes did vary by company size, but a prevailing concern across all companies related to questions about the efficiency/effectiveness of contact with vocational rehabilitation agencies. For both small- and mid-sized companies, there was a belief that people with disabilities could not do the work or were somehow less qualified. For large companies, convincing departmental and team managers that outreaching workers with disabilities would be a worthwhile hiring practice remained a challenge. Conclusion The themes derived from this study can be used to help occupational rehabilitation professionals develop educational and marketing interventions to improve employers’ attitudes toward hiring and retaining individuals with disabilities.     

Clinical features and treatment outcomes of vancomycin-intermediate <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> (VISA) and heteroresistant vancomycin-intermediate <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> (hVISA) in a tertiary care institution in Singapore

This retrospective case–control study was undertaken to review the clinical features associated with heteroresistant vancomycin-intermediate Staphylococcus aureus (hVISA) and vancomycin-intermediate S. aureus (VISA) infections and the local impact they have on clinical outcome. Compared with vancomycin-susceptible S. aureus (n = 30), hVISA and VISA infections (n = 10) are found to be associated with a longer period of prior glycopeptide use (P = 0.01), bone/joint (P < 0.01) and prosthetic infections (P = 0.04), as well as treatment failure, as evidenced by longer bacteremic (P < 0.01) and culture positivity (P < 0.01) periods. This was observed to have resulted in longer hospital length of stay (P < 0.01) and total antibiotic therapy duration (P = 0.01). There was, however, no significant difference in the overall patient mortality or the hospitalization cost (P = 0.12) in both groups. Clinicians should be cognizant of the association between hVISA/VISA with high bacterial load deep-seated infections. We recommend targeted and even universal screening for hVISA/VISA in methicillin-resistant S. aureus (MRSA) infections.     

Combined transfection with EBV-specific epitopes and HLA-A2 genes is more effective than separate transfection in promoting CTL lysis against nasopharyngeal carcinoma

To augment specific cytotoxic T lymphocyte(CTL)lysis is a promising strategy for cancer therapy,in this study,we examined the boosting effect of CTLs upon autologous lymphoblastoid B cell lines(LCLs)transfected withdiverse plasmids,to explore the possible CTL-based immunotherapy of nasopharyngeal carcinoma(NPC).FCM analysis displayed rather high ratio(>30%)of successfully transfected LCLs by utilizing the DMRIE-Ckit.CTL assays demonstrated that substantially higher ratio of CTL specific lysis was observed upon the LCLstransfected with both expression vectors encoding EBV-specific epitopes and their presentation moleculeHLA-A2,in contrast with those transfected separately.By transfecting the vector encoding HLA-A2 alone,onlythe LCLs of HLA-A2~+ donors elicited markedly higher CTL lysis.CTL assays also showed that there existed nomarked differences upon transfection by either different vectors(pcDNA3,pNGVL3 or pNGVL3-hFlex),ordifferent EBV-derived peptides(LMP_2Pepl or LMP_2Pep2),or with or without the doubled DNA sequenceencoding peptides.This study indicated a promising immunotherapy strategy on NPC through boosting andeliciting the EBV-specific CTL activation by transferring vectors encoding both EBV-specific epitopes and theirpresentation molecule HLA-A2 into autologous LCL,the presentation cells of MHC/peptide tetramericcomplex.Cellular & Molecular Immunology.2004;1(3):229-234.     

Single-nucleotide polymorphisms in the KCNN3 gene associate with preterm birth

The objectives were to determine whether single-nucleotide polymorphisms (SNPs) in KCNN3 (encodes the small conductance calcium-activated potassium channel subfamily N, member 3), associate with preterm birth (PTB). In all, 602 preterm families with at least 1 preterm (<37 weeks gestation) infant were studied: DNA from the infant and one or both parents were genotyped for 16 SNPs in KCNN3. A region of interest within KCNN3 was sequenced in 512 Caucasian non-Hispanic mothers (412 with preterm deliveries;100 who delivered at term). Family-based association testing was used for genotyping analysis; Fisher exact test was used for sequencing analysis. Six SNPs (rs1218585, rs4845396, rs12058931, rs1218568, rs6426985, and rs4845394) were associated with PTB (all Ps < .05). These variations were all located within the intronic region between exons 1 and 2. Maternal sequencing revealed an association of 3 SNPs with spontaneous PTB; rs1218585 (P = .007), rs1218584 (P = .05), and a novel SNP at chromosome1:153099353 (P = .02). Polymorphisms in KCNN3 are associated with PTB and investigation into the functional significance of these allelic changes is warranted.