Targeting enhancer of zeste homolog 2 protects against acute kidney injury.

Despite the established oncogenic and profibrotic functions of enhancer of zeste homolog 2 (EZH2), a methyltransferase that induces histone H3 lysine 27 trimethylation (H3K27me3), its role in acute kidney injury (AKI) remains unclear. In this study, we demonstrated that EZH2 and H3K27me3 were upregulated in the murine kidney with AKI induced by either ischemia-reperfusion (I/R) or folic acid (FA). Pharmacologic inhibition of EZH2 with 3-deazaneplanocin A (3-DZNeP) prevented tubular injury in both models as demonstrated by reduced renal dysfunction, diminished neutrophil gelatinase-associated lipocalin expression and decreased renal tubular cell death. Injury to the kidney resulted in reduced expression of E-cadherin and ZO-1, whereas EZH2 inhibition largely preserved their expression. Moreover, 3-DZNep was effective in counteracting the increased expression of matrix metalloproteinase (MMP)-2 and MMP-9, as well as the phosphorylation of Raf-1 and ERK1/2 in the injured kidney. Conversely, blocking EZH2 reversed the decrease of tissue inhibitor of metalloproteinase (TIMP)-2 and metalloproteinase (TIMP)-3, and Raf kinase inhibitor protein (RKIP) in the kidney after acute injury. Similarly, oxidant injury to cultured kidney proximal tubular epithelial cells caused a decrease in the expression of E-cadherin, ZO-1, TIMP-2/-3, and RKIP, as well as an increase in the expression of MMP-2/9 and phosphorylation of Raf-1 ERK1/2. Blocking EZH2 with 3-DZNep or SiRNA hindered these responses. Thus, these results suggest that targeting EZH2 protects against AKI through a mechanism associated with the preservation of adhesion/junctions, reduction of matrix metalloproteinases and attenuation of the Raf-1/ERK1/2 pathway.

Head and face injuries in scholastic women's lacrosse with and without eyewear.

PURPOSE: The use of protective equipment has been absent or inconsistent in scholastic women's lacrosse leading to increasing concern for eye and head injury. There is a paucity of field data, however, on which to base strategic decisions on how best to prevent head injuries in young athletes. METHODS: This study examined the effects of protective eyewear on injury rates in scholastic women's lacrosse in a cohort of approximately 700 varsity and junior varsity players in central New York studied prospectively for 2 yr during a transition from sparse to almost complete eyewear use. RESULTS: The overall head/face injury rate was 0.71 injuries per 1000 exposures (games and practices) and was 16.5% lower in goggle wearers. In games alone, where more aggressive play and stick use prevails, the rate associated with protective eyewear use was markedly lower (51%). Considering specific regions, the rates for peri-orbit and forehead injuries among goggle users were substantially lower than for nonusers (6% and 13%, respectively). Cheek and scalp injury rates tended to be higher among goggle wearers, but not statistically significantly so. Significant compensatory increases with goggle wear at other sites were not observed. Only a few injuries appeared to be mediated by the goggles themselves and potentially could have been more serious if the goggles had not been present. No direct eye (orbit) injuries were reported throughout the study period. CONCLUSION: On balance, then, the use of eyewear in women's lacrosse appears to be beneficial when users are compared with nonusers.

Phenotypic and functional similarities between 5-azacytidine-treated T cells and a T cell subset in patients with active systemic lupus erythematosus.

OBJECTIVE: Antigen-specific CD4+ T cells treated with DNA methylation inhibitors become autoreactive, suggesting a novel mechanism for autoimmunity. To test whether this mechanism might be involved in systemic lupus erythematosus (SLE), phenotypic markers for the autoreactive cells were sought. METHODS: Cloned normal T cells were treated with the DNA methylation inhibitor 5-azacytidine (5-azaC) and studied for altered gene expression. T cells from patients with active SLE were then studied for a similar change in gene expression, and cells expressing the marker were tested for autoreactivity. RESULTS: 5-azaC-treated normal T cells had increased CD11a (leukocyte function-associated antigen 1 alpha) expression relative to other membrane molecules. A T cell subset with similar CD11a expression was found in patients with active SLE. This subset contained cells that spontaneously lysed autologous macrophages, with a specificity similar to that of 5-azaC-treated cells.

Detection of HIV-1 antibodies in blood specimens spotted on filter-paper.

Described are the results of an international collaborative study to evaluate the use of whole blood samples spotted on filter-paper (BSP) for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). BSP samples were collected from 40 patients at risk for HIV-1 infection and tested blindly using commercially available HIV antibody test kits, either specifically manufactured or modified for this purpose. Parallel serum samples were also collected, and the antibody reactivity was defined and confirmed by Western blot. The results demonstrate that recovery of antibodies from BSP samples after elution can be comparable to that from serum. Some kits can be easily adapted to test BSP samples, while others cannot. At present, detection of HIV antibodies in BSP samples should therefore be carried out using kits specifically manufactured for this purpose or by the development of a modified protocol using a panel of BSP and their corresponding serum specimens.

Immunosuppressive drug therapy for rheumatic disease.

The use and complications of intravenous cyclophosphamide, azathioprine, nitrogen mustard, and cyclosporine are reviewed. Studies have failed to demonstrate superiority of intravenous versus oral cyclophosphamide in rheumatic diseases other than systemic lupus. Azathioprine is a useful immunosuppressive agent in Beçhet's disease. Further studies of cyclosporine in rheumatoid arthritis indicate that efficacy and toxicity are both dose related.

Cultivation of human immunodeficiency virus from whole blood: effect of anticoagulant and inoculum size on virus growth.

Human immunodeficiency virus (HIV) was cultivated directly from whole blood treated with anticoagulant by cocultivation with phytohaemagglutinin-stimulated cord blood lymphocytes. When heparin was used as the anticoagulant, isolation rates were low (10% to 56%, depending on the patient group); but when EDTA was used, isolation rates were much higher (50% to 100%). Culture of whole blood gave results identical to those of culture of separated peripheral mononuclear cells, and in some cases virus could be isolated from as little as 10 microliters of unseparated EDTA anticoagulated blood.

The prevalence of serological markers for hepatitis B virus infection in Australian Naval personnel.

In order to determine the prevalence of hepatitis B virus infection in Royal Australian Navy personnel and to formulate policies for their education, screening and vaccination, a voluntary survey was undertaken. Seven hundred and forty-eight subjects completed a questionnaire that provided details of age, sex, rank, ethnic origin, service abroad, length of service, history of tattooing, liver disease and blood transfusions. Serum was assayed by radioimmunoassay for the presence of hepatitis B surface antigen (HBsAg), antibody to hepatitis B surface antigen (antiHBs) and antibody to hepatitis B core antigen (antiHBc). The presence of markers of hepatitis B virus infection was correlated with "risk factors" by means of the chi 2 test of independence. Fourteen (74%) of 19 personnel from the Pacific Islands, Asia and Africa who were excluded from analysis showed one-or-more such markers. The prevalence of any marker of hepatitis B virus infection was 4.4% (95% confidence interval [CI], 2.9%-5.9%). The individual markers HBsAg, antiHBs and antiHBc were found in 0.4% (95% CI, 0.1%-1.2%), 3.6% (95% CI, 2.2%-4.9%) and 3.9% (95% CI, 2.5%-5.3%) of subjects, respectively. The annual attack rate was estimated to be less than 1%. Factors which had a significant correlation with an increased prevalence of markers of hepatitis B virus infection included tattooing (especially if this had been performed in Asia), age, the duration of service and service abroad. Based on these data, and given the reduced cost of hepatitis B vaccines, vaccination now is considered worth while. A policy of vaccinating all personnel who may be required for service at sea, and others at risk, now is in effect. Educational programmes that are aimed at minimizing the risk of exposure to hepatitis B and other viral infections have been instituted.

The regulated expression of Epstein-Barr virus: evidence that the transition from primary to secondary protein synthesis in Raji cells superinfected with Epstein-Barr virus requires the synthesis of new RNA.

In the presence of canavanine and the absence of arginine, superinfected Raji cells synthesize a limited spectrum of proteins (primary proteins). After the removal of canavanine at 8 h post infection, the cells proceed after a lag phase of 2-3 h to synthesize a second group of proteins. The appearance of these proteins can be prevented by addition of actinomycin-D to the chase media, suggesting that active primary proteins are required for the synthesis of new mRNA coding for the secondary proteins.

Ultrastructural studies on cell fusion induced by Epstein-Barr virus or N-butyrate and 12-O-tetradecanoylphorbol-13-acetate. Brief report.

The morphological changes which occur in Raji cells during EBV induced fusion are described. Of particular interest is the formation of local contacts between cells, at these points the plasmalemmae of the two cells become disorganized and cytoplasmic bridges are formed.

Strategies for the economic preparation of Epstein-Barr virus proteins of diagnostic and protective value by genetic engineering: a new approach based on segments of virus-encoded gene products.

Immunoprecipitation of Epstein-Barr viral proteins with various sera from normal adults, patients with fresh infectious mononucleosis or nasopharyngeal carcinoma was used to identify antigens which are of importance in the determination of immune status and characteristic of a particular disease. Some genes coding for of these antigens have been localized on the Epstein-Barr virus (EBV) genome by hybrid-selected translation. With the use of sequence data, these genes could then be subcloned from EBV DNA and expressed in eukaryotic and prokaryotic cells. Data on the expression are presented and the application of the methods described for the production of diagnostic reagents and vaccines is discussed.