Transforming growth factor-β enhances secretory component and major histocompatibility complex class I antigen expression on rat IEC-6 intestinal epithelial cells

Transforming growth factor-β (TGF-β) has been implicated as having a role in inflammatory responses by inducing cellular infiltration and the release of inflammatory cytokines. In this study, the IEC-6 rat intestinal epithelial cell line was used as a model to assess the effect of TGF-β₁ on the expression of various plasma membrane determinants. TGF-β₁ induced a dose-dependent increase in the percentage of cells expressing surface secretory component (SC) and class I major histocompatibility (MHC) antigens. However, the expression of class II MHC was unaffected. In contrast, epidermal growth factor had no effect on any of the surface proteins studied. The TGF-β₁-enhanced expression of SC was accompanied by an enhanced binding of polymeric, but not monomeric, immunoglobulin A (IgA). Preincubation of the TGF-β₁-treated cells with an anti-human β-galactosyltransferase (β-GT) antiserum did not block the binding of the anti-SC antibody, indicating that the TGF-β-induced increase in SC staining was due to SC expression and not the polymeric immunoglobulin-binding enzyme, β-GT. These results indicate that TGF-β₁ may be important in immune functions involving intestinal epithelial cells by enhancing the expression of surface class I MHC antigens and SC, a protein responsible for the transport of polymeric IgA into the intestinal lumen.     

CpG methylation controls reactivation of HIV from latency

What are the purposes of prizes and recognitions? Are they lagging indicators of past achievements or leading indicators of things to come?     

Mislocalization of the MRN complex prevents ATR signaling during adenovirus infection

The protein kinases ataxia‐telangiectasia mutated (ATM) and ATM‐Rad3 related (ATR) are activated in response to DNA damage, genotoxic stress and virus infections. Here we show that during infection with wild‐type adenovirus, ATR and its cofactors RPA32, ATRIP and TopBP1 accumulate at viral replication centres, but there is minimal ATR activation. We show that the Mre11/Rad50/Nbs1 (MRN) complex is recruited to viral centres only during infection with adenoviruses lacking the early region E4 and ATR signaling is activated. This suggests a novel requirement for the MRN complex in ATR activation during virus infection, which is independent of Mre11 nuclease activity and recruitment of RPA/ATR/ATRIP/TopBP1. Unlike other damage scenarios, we found that ATM and ATR signaling are not dependent on each other during infection. We identify a region of the viral E4orf3 protein responsible for immobilization of the MRN complex and show that this prevents ATR signaling during adenovirus infection. We propose that immobilization of the MRN damage sensor by E4orf3 protein prevents recognition of viral genomes and blocks detrimental aspects of checkpoint signaling during virus infection.     

Stable isotope-coded quaternization for comparative quantification of estrogen metabolites by high-performance liquid chromatography-electrospray ionization mass spectrometry

A fast and sensitive LC-ESI-MS method is described for the comparative quantification of 16 estrogen metabolites based on the derivatization of estrogens with a novel derivatizing reagent, N-methyl-nicotinic acid N-hydroxysuccinimide ester (C1-NA-NHS). The process introduces a quaternary amine to the analytes, making the analytes permanently charged regardless of the pH of the high-performance liquid chromatography (HPLC) mobile phase. This quaternization resulted in a highly efficient separation of 16 estrogen metabolites in 7 min at a detection level below 1 ng/mL. By using a deuterated derivatizing reagent (C1-d(3)-NA-NHS), a complete set of deuterated standards was utilized and used as internal standards in a comparative quantification and recovery study, demonstrating acceptable results over a wide concentration range. A pooled breast cancer serum sample was analyzed using the described method, and 15 estrogens were detected in the range of 80-530 pg/mL.     

alpha-Lipoic acid modulates extracellular matrix and angiogenesis gene expression in non-healing wounds treated with hyperbaric oxygen therapy

alpha-Lipoic acid (LA) has been found previously to accelerate wound repair in patients affected by chronic wounds who underwent hyperbaric oxygen (HBO) therapy. Because proteinases are important in wound repair, we hypothesized that LA may regulate matrix metalloproteinase (MMP) expression in cells that are involved in wound repair. Patients undergoing HBO therapy were double-blind randomized into two groups: the LA group and the placebo group. Gene expression profiles for MMPs and for angiogenesis mediators were evaluated in biopsies collected at the first HBO session, at the seventh HBO session, and after 14 days of HBO treatment. ELISA tests were used to validate microarray expression of selected genes. LA supplementation in combination with HBO therapy downregulated the inflammatory cytokines and the growth factors which, in turn, affect MMPs expression. The disruption of the positive autocrine feedback loops that maintain the chronic wound state promotes progression of the healing process.     

Implementation of contact definitions calculated by FEA to describe the healing process of basal implants

Aims: Bone structure around basal implants shows a dual healing mode: direct contact areas manifest primary osteonal remodeling, in the void osteotomy-induced spaces, the repair begins with woven bone formation. This woven bone is later converted into osteonal bone. The purpose of this study was to develop a model to accurately represent the interface between bone and basal implant throughout the healing process. The model was applied to the biological scenario of changing load distribution in a basal implant system over time. Methods: Computations were made through finite element analysis using multiple models with changing boneimplant contact definitions which reflected the dynamic nature of the interface throughout the bony healing process. Five stages of bony healing were calculated taking into account the changes in mineral content of bone in the vicinity of the load transmitting implant surfaces. Results: As the bony integration of basal implants proceeds during healing, peak stresses within the metal structure shift geographically. While bony repair may still weaken osteonal bone, woven bone has already matured. This leads to changes in the load distribution between and within the direct contact areas, and bone areas which make later contact with implant. Conclusions: This study shows that basal implants undergo an intrinsic shift of maximum stress regions during osseointegration. Fatigue testing methods in the case of basal implants must therefore take into account this gradual shift from early healing phase until full osseointegration is achieved.     

Poor survival of ABG I hip prosthesis in younger patients

Background: Hydroxyapatite coated (HAC) hip implants have been used in clinical practice for more than two decades. However, the majority of studies have reported only intermediate term outcomes that are not reliable for predicting long-term behavior in all implants. The aim of this study was to determine the performance of HAC total hip arthroplasty in younger patients over a 10-year follow-up period. Methods and Results: This was an observational retrospective study of a 137 consecutive hips with the ABG I prosthesis. Of these, 128 were available for the last investigation. Median duration of follow-up was 10.9 years. The mean age at time of index surgery was 46+/-6.7 years. Probability of implant survival was estimated using the Kaplan-Meier method. The overall 12-year cumulative survival was 0.55 (95% CI, 0.443-0.659). Periprosthetic osteolysis (57 %) was the most frequent reason for failure followed by aseptic loosening (28 %). When only aseptic loosening was included in the analysis, the same figures for cup and stem were 0.873 (95% CI, 0.808-0.938) and 0.992 (95% CI, 0.976- 1.0), respectively. Patients with a smaller cup size were those at high risk for revision due to wear-related complications (odds ratio, OR=4.3; 95% CI, 1.734-10.555). Conclusion: This study reports one of the poorest 12-year survivorship data for cementless acetabular component in the literature. The main reason for premature failure was osteolysis, strongly related to high wear rate of polyethylene.     

The experimental contamination of foodstuffs with the spores of toxigenic micromycetes and the production of mycotoxins

A study of experimental contamination of foods with spores of toxigenic micromycetes (Aspergillus flavus andPenicillium verrucosum) was realized. Samples of bread, apricot jam, Edam cheese, processed cheese and half-fat dried milk purchased on the market were used for experimental contamination. Aflatoxins and ochratoxin A were determinated in experimental contaminated foodstuffs.     

The mycotoxin research in foodstuffs in the Czech Republic in the 90th Years

Mycotoxins are naturally occurring secondary metabolites produced by several toxigenic microscopic fungi on a variety of crops, especially cereal grains and further foodstuffs. Series of experimental research projects on the determination of mycotoxins (aflatoxins, cyclopiazonic acid, ochratoxin A, patulin, deoxynivalenol, fumonisin B₁, T-2 toxin, zearalenone, sterigmatocystin, alternaria toxins) in several foods were realized in the National Reference Centre for Microscopic Fungi and Mycotoxins in the 90^th years. The aim of our work was an estimation of dietary exposure to mycotoxins and risk assessment. The method of a solid phase extraction (SPE), liquid — liquid extraction and immunoaffinity chromatography (f. e. R-Biopharm, VICAM) were used to elaborate for sample analyses of mycotoxins in our projects. The mycotoxins were detected most frequently by chromatographic methods (HPTLC, HPLC, GC) and immunochemical methods (ELISA). Average dietary exposure has been calculated by multiplying of concentration data for specific foods with their consumption rates per 1 kg of b. w. per day. The estimation of the dietary exposure dose of mycotoxins for the Czech population is presented.