Normal colon tissue and colon carcinoma show no difference in heparanase promoter methylation

Introduction

Heparanase, the sole heparan sulfate degrading enzyme, has a role in cellular invasion. Accordingly, a large number of studies have demonstrated an association between heparanase expression and tumor stage and patients' prognosis. In colon carcinoma, heparanase shows increased expression in tumor compared to normal tissue and its expression correlates with the presence of metastasis. One of the regulatory mechanisms of heparanase expression is de-methylation on its promoter. In the present study we evaluated the role of heparanase promoter methylation in colon carcinoma.

Material and methods

Analysis of heparanase promoter methylation was done on 32 samples of colon carcinoma as well as 30 samples of normal colonic mucosa. DNA was extracted from FFPE tissue and subjected to bisulfite conversion. The relative fraction of methylated and unmethylated DNA was evaluated using quantitative real-time PCR.

Results

The fraction of methylated DNA was 1 ± 3.4% in the colon carcinoma group, and 2.5 ± 3.3% in the normal colon group (P = 0.11). Only one case in the normal group and one case in the tumor group showed more than 10% methylation in the heparanase promoter.

Conclusion

We did not find any significant difference in heparanase promoter methylation between colon carcinoma and normal colonic mucosa, suggesting that heparanase overexpression in colon carcinoma is mediated by other mechanisms.     

The inhibitory receptor CD300a is essential for neutrophil-mediated clearance of urinary tract infection in mice

Neutrophils play a crucial role in immune defense against and clearance of uropathogenic Escherichia coli (UPEC)-mediated urinary tract infection, the most common bacterial infection in healthy humans. CD300a is an inhibitory receptor that binds phosphatidylserine and phosphatidylethanolamine, presented on the membranes of apoptotic cells. CD300a binding to phosphatidylserine and phosphatidylethanolamine, also known as the “eat me” signal, mediates immune tolerance to dying cells. Here, we demonstrate for the first time that CD300a plays an important role in the neutrophil-mediated immune response to UPEC-induced urinary tract infection. We show that CD300a-deficient neutrophils have impaired phagocytic abilities and despite their increased accumulation at the site of infection, they are unable to reduce bacterial burden in the bladder, which results in significant exacerbation of infection and worse host outcome. Finally, we demonstrate that UPEC's pore forming toxin α-hemolysin induces upregulation of the CD300a ligand on infected bladder epithelial cells, signaling to neutrophils to be cleared.     

Characterization of the expression of MHC proteins in human embryonic stem cells

Human embryonic stem (ES) cells are pluripotent cells that may be used in transplantation medicine. These cells can be induced to differentiate into cells from the three embryonic germ layers both in vivo and in vitro. To determine whether human ES cells might be rejected after transplantation, we examined cell surface expression of the MHC proteins in these cells. Our results show very low expression levels of MHC class I (MHC-I) proteins on the surface of human ES cells that moderately increase on in vitro or in vivo differentiation. A dramatic induction of MHC-I proteins was observed when the cells were treated with IFN-gamma but not with IFN-alpha or -beta. However, all three IFNs induced expression of MHC-I proteins in differentiated human ES cells. MHC-II proteins and HLA-G were not expressed on the surface of undifferentiated or differentiated cells. Ligands for natural killer cell receptors were either absent or expressed in very low levels in human ES cells and in their differentiated derivatives. In accordance, natural killer cytotoxic assays demonstrated only limited lysis of both undifferentiated and differentiated cells. To initiate a histocompatibility databank of human ES cells, we have isotyped several of the published ES cell lines for their human leukocyte antigens. In conclusion, our results demonstrate that human ES cells can express high levels of MHC-I proteins and thus may be rejected on transplantation.     

Establishing a multidisciplinary diabetic foot team in a large tertiary hospital: a workshop

Every year, over 1 million people with diabetes lose a leg due to diabetic foot disease. Most amputations are preceded by a foot ulcer. Causes for the development of foot ulcers are generally multifactorial and may include neuropathy, peripheral vasculopathy, abnormal foot mechanics and infection. Multidisciplinary approach to the patient with acute diabetic foot is mandatory and has been shown to reduce amputation rate. In our article we describe the establishment of a multidisciplinary diabetic foot team in a large tertiary hospital and its outcomes. Copyright © 2014 John Wiley & Sons, Ltd.     

The Influence of Residual Alveolar Bone Height on Graft Composition after Maxillary Sinus Augmentation Using Two Different Xenografts: A Histomorphometric Comparative Study

Aim: To evaluate the hypothesis of a correlation between the preoperative residual alveolar bone height (RBH) and graft maturation after maxillary sinus floor augmentation procedures using two different bone substitutes. Methods: A total of 20 patients who underwent unilateral maxillary sinus floor augmentation with either mineralized deproteinized bovine bone (DBBM) or a xenograft enriched with polymer and gelatin (NBS) were included in this prospective study. Six months after sinus surgery, bone biopsies were harvested with a 3.2 mm diameter trephine bur, prior to dental implant placement. Histomorphometric analysis was performed, and the results were correlated with the individual RBH. Implants were loaded after 5 months of insertion, and 1-year implant success and marginal bone level change were assessed. Results: RBH was 2.17 ± 1.11 mm (range 0.5–3.5 mm) and 2.14 ± 0.72 mm (range 0.5–3.0 mm) in the NBS and DBBM group, respectively. The biopsy analyses for the DBBM group showed woven bone increases by 5.08% per 1-mm increment of RBH; medullary spaces decreased by 9.02%, osteoid decreased by 4.4%, residual biomaterial decreased by 0.34%, and lamellar bone increased by 5.68% per 1-mm increase of RBH. In the NBS group, samples showed woven bone increases by 8.08% per 1-mm increase of RBH; medullary spaces decreased by 0.38%; osteoid increased by 1.34%, residual biomaterial decreased by 0.58%, and lamellar bone decreased by 5.50% per 1-mm increase of RBH. There was no statistically significant difference in the correlation between RBH and lamellar bone, woven bone, and osteoid, independently of the material used. Implant success was 100% in both groups, and marginal bone loss was 1.02 ± 0.42 mm in DBBM and 0.95 ± 0.31 mm in the NBS group after the 1-year follow-up. Conclusion: In spite of the absence of significance, the observed trend for woven bone to increase and medullary spaces to decrease when RBH increases deserves attention. Residual bone dimension might be a determinant in the bone graft maturation after maxillary sinus augmentation.     

Epstein–Barr virus antibodies mark systemic lupus erythematosus and scleroderma patients negative for anti‐DNA

Systemic lupus erythematosus (SLE) is an autoimmune disease that can attack many different body organs; the triggering event is unknown. SLE has been associated with more than 100 different autoantibody reactivities – anti‐dsDNA is prominent. Nevertheless, autoantibodies to dsDNA occur in only two‐thirds of SLE patients. We previously reported the use of an antigen microarray to characterize SLE serology. We now report the results of an expanded study of serology in SLE patients and scleroderma (SSc) patients compared with healthy controls. The analysis validated and extended previous findings: two‐thirds of SLE patients reacted to a large spectrum of self‐molecules that overlapped with their reactivity to dsDNA; moreover, some SLE patients manifested a deficiency of natural IgM autoantibodies. Most significant was the finding that many SLE patients who were negative for autoantibodies to dsDNA manifested abnormal antibody responses to Epstein–Barr virus (EBV): these subjects made IgG antibodies to EBV antigens to which healthy subjects did not respond or they failed to make antibodies to EBV antigens to which healthy subjects did respond. This observation suggests that SLE may be associated with a defective immune response to EBV. The SSc patients shared many of these serological abnormalities with SLE patients, but differed from them in increased IgG autoantibodies to topoisomerase and centromere B; 84% of SLE patients and 58% of SSc patients could be detected by their abnormal antibodies to EBV. Hence an aberrant immune response to a ubiquitous viral infection such as EBV might set the stage for an autoimmune disease.