A novel monoclonal antibody recognizing β(1–3) glucans in intact cells of Candida and CryptococcusKONDORI, N.; EDEBO, L.; MATTSBY‐BALTZER, I.
doi: 10.1111/j.1600-0463.2008.01013.xpmid: 19132980
The cell walls of all medically important fungi contain a unique polyglucose compound, β(1–3) glucan. In the present study, murine monoclonal antibodies were produced against linear and β(1–6) branched β(1–3) glucans, and their specificities were characterized for reactivity to other β glucans, fungal cell wall fragments, and fungal cells. Their reactivity was also compared with that of rabbit polyclonal antibodies raised against the same immunogens. Two mouse monoclonal antibodies (AG and BG) recognized immunoreactive epitopes in β(1–3)(1–6) glucan by ELISA. In an inhibition assay of the anti‐β(1–3)(1–6) activity of the monoclonals, the homologous antigen effectively inhibited the activity as expected, while β(1–3) also inhibited the assay but to a much lesser extent. No inhibition was obtained by β(1–3)(1–4) or β(1–6), while a cell wall extract of Candida albicans (PPM) effectively inhibited both monoclonals. Cell wall fragments of C. albicans (CaCW) and Cryptococcus neoformans (CnCW) inhibited the anti‐β(1–3)(1–6) activity of AG, while BG was much less or not inhibited at all. Immunofluorescence confirmed the unique antibody specificity of AG by its recognition of a β(1–3)(1–6)‐associated epitope on the cell surfaces of C. albicans,C. krusei, C. glabrata, and nonencapsulated C. neoformans. The epitope for the AG antibody is suggested to be present in the branching point of β(1–3)(1–6), or in the randomly coiled β(1–3) polyglucan due to the presence of branches. Thus, monoclonal antibodies to β(1–3)(1–6) glucans may have potential as tools in the laboratory diagnosis of invasive yeast infections.
Clinical isolates of Staphylococcus aureus from the Arkhangelsk region, Russia: antimicrobial susceptibility, molecular epidemiology, and distribution of Panton‐Valentine leucocidin genesVOROBIEVA, V.; BAZHUKOVA, T.; HANSSEN, A. M.; CAUGANT, D. A.; SEMENOVA, N.; HALDORSEN, B. C.; SIMONSEN, G. S.; SUNDSFJORD, A.
doi: 10.1111/j.1600-0463.2008.01092.xpmid: 19132981
A total of 91 consecutive clinical isolates of Staphylococcus aureus were collected at the Regional Hospital of Arkhangelsk, Russia, from May to December 2004, and examined for antimicrobial susceptibility, methicillin resistance and presence of Panton‐Valentine leucocidin (PVL) genes. Epidemiological typing was performed by pulsed‐field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Methicillin‐resistant S. aureus (MRSA) isolates were examined by staphylococcal cassette chromosome mec (SCCmec) typing. High‐to‐moderate rates of resistance to penicillin (β‐lactamase production; 93%), tetracycline (40%), erythromycin and clindamycin (32%) were observed. Forty out of ninety‐one (44%) isolates were positive for PVL genes. Thirty‐six (40%) PVL‐positive methicillin‐susceptible S. aureus (MSSA) strains were shown by PFGE and MLST typing (ST121, ST681, ST837) to be part of a nosocomial outbreak caused by clonal complex (CC) 121. PFGE, MLST and SCCmec typing revealed three MRSA clones. Sequence type (ST) 239‐III (n=11), ST1097‐III (n=1) and ST8‐IV (n=3) belong to CC8 of epidemic multiresistant MRSA, whereas ST426‐MRSA‐IV/CC395 (n=1) has not been reported previously. All MRSA strains were PVL negative. The overall results underline the necessity of microbiological sampling, antimicrobial susceptibility testing, and epidemiological typing as a rational basis for antimicrobial treatment of S. aureus infections, and infection control measures to limit the spread of multiresistant MRSA and epidemic MSSA clones.
Heat shock proteins 27, 60 and 70 as prognostic markers of prostate cancerGLAESSGEN, AXEL; JONMARKER, SARA; LINDBERG, ANNA; NILSSON, BO; LEWENSOHN, ROLF; EKMAN, PETER; VALDMAN, ALEXANDER; EGEVAD, LARS
doi: 10.1111/j.1600-0463.2008.01051.xpmid: 19132982
Heat shock proteins (HSPs) protect cells against stress‐associated injury and are overexpressed in several malignant tumors. We aimed to investigate their value as prognostic markers in prostate cancer. A tissue microarray (TMA) was constructed of 289 prostate cancers from radical prostatectomy (RP) specimens with median follow‐up of 48.9 months. Slides were immunostained for HSP27, HSP60 and HSP70. Intensity and extent of immunoreactivity (IR) and their product (IRp) was evaluated by two observers. The IRp of HSP27 and HSP60, but not of HSP70, significantly predicted biochemical recurrence (p=0.014, 0.034 and 0.160, respectively). Recurrence‐free survival in patients with strong HSP27 and HSP60 staining was shorter than in those with weak expression (p=0.019 and 0.001, respectively). IRp of HSP27 and HSP60 correlated with Gleason score (p<0.01). HSP60 was an independent predictor of biochemical recurrence in multivariate analysis, including extraprostatic extension, margin status, seminal vesicle invasion and Gleason score. Weighted kappa for interobserver agreement of HSP27, HSP60 and HSP70 IR was 0.613–0.823 for intensity and 0.584–0.719 for IRp, but only 0.036–0.244 for extent, raising the question whether staining extent should be estimated on TMA. We conclude that HSP27 and HSP60 are predictors of biochemical recurrence after RP.
Higher FoxP3 mRNA expression in peripheral blood mononuclear cells of GAD65 or IA‐2 autoantibody‐positive compared with autoantibody‐negative personsLINK, MAIRE; SALUR, LIINA; RAJASALU, KAI KISAND; TILLMANN, VALLO; UIBO, RAIVO
doi: 10.1111/j.1600-0463.2008.00889.xpmid: 19132983
The role of regulatory T cells (Tregs) in type 1 diabetes has been studied extensively. The most prevalent way to define Tregs has been by their surface expression of CD4 and CD25. As currently the transcription factor FoxP3 and the low expression of CD127 are regarded to be the most specific markers of Tregs, we analysed the number of Tregs defined by these molecules in peripheral blood mononuclear cells of diabetic patients and healthy controls. The gene expression of transforming growth factor β and two isoforms of FoxP3 was measured as well. There were no significant differences between diabetic patients and healthy controls regarding the number of Tregs, or the expression of FoxP3 isoforms and TGFβ in peripheral blood mononuclear cells. However, we found significantly higher expression of both full‐length and Δ2FoxP3 in study subjects, positive for either GAD65 or IA‐2 autoantibodies. The ratio of the expression of different isoforms was not changed. This study shows the possible role of FoxP3 in the development of tissue characteristic humoral immunity in type 1 diabetes.
Stereological quantification of tumor volume, mean nuclear volume and total number of melanoma cells correlated with morbidity and mortalityBØNNELYKKE‐BEHRNDTZ, MARIE LOUISE; SØRENSEN, FLEMMING BRANDT; DAMSGAARD, TINE ENGBERG
doi: 10.1111/j.1600-0463.2008.01100.xpmid: 19132984
Stereological quantification of tumor volume, total number of tumor cells and mean nuclear volume provides unbiased data, regardless of the three‐dimensional shape of the melanocytic lesion. The aim of the present study was to investigate whether these variables are reproducible and may represent potential indicators of prognosis. Sixty patients who underwent surgery at the Department of Plastic Surgery, Aarhus University Hospital, from 1991 to 1994 were included in the study. Total tumor volume was estimated by the Cavalieri technique, total number of tumor cells by the optical dissector principle, and nuclear mean volume by point counting. Tumor volume, total number of tumor cells and nuclear mean volume were all reproducible parameters, showing a non‐significant intra‐observer variation (p=0.72; p=0.83, and p=0.15). Grouped according to the median values, the disease‐free survival time was significantly different for tumor volume as well as total number of tumor cells (p=0.0006 and p=0.0004), but no significant difference in mortality was found (p=0.1 and p=0.06). The mean nuclear volume showed no difference regarding disease‐free survival or mortality (p=0.5 and p=0.8). Breslow's thickness showed a significant impact on both disease‐free survival (p=0.001) and mortality (p=0.009). In conclusion, tumor volume and total number of cancer cells were highly reproducible but did not add additional, independent prognostic information regarding the study population.
Prognostic value of COX‐2 immunohistochemical expression evaluated by quantitative image analysis in colorectal cancerZAFIRELLIS, KYRIAKOS; AGROGIANNIS, GEORGE; ZACHAKI, AGLAIA
doi: 10.1111/j.1600-0463.2008.01104.xpmid: 19132985
Epidemiological studies have shown that the inducible form of cyclooxygenase (COX‐2) may be involved in colorectal carcinogenesis, but it is controversial whether its expression is a prognostic factor for colorectal cancer. The aim of the study was to examine the expression of COX‐2 in colorectal cancer and investigate its prognostic relevance. Tissue sections of primary tumors from 132 patients undergoing curative resection for colorectal cancer were immunohistochemically examined for COX‐2 expression. The levels of intensity and extent of COX‐2 staining were quantified by use of a computerized image analysis system and correlated with various clinicopathological characteristics and survival. COX‐2 immunoreactivity was observed in the cytoplasm of tumour epithelial cells of all colorectal cancer tissues examined. No significant correlation was found between levels of intensity and extent of COX‐2 staining and various clinicopathological characteristics, including age, gender, tumor location, tumor size, tumor grade, depth of invasion, lymph node status and TNM stage. There was an inverse correlation between intensity and extent of COX‐2 staining scores (Spearman's rho=−0.414; p<0.001). To analyze the prognostic value of intensity and extent of COX‐2 staining, the patients were divided into four groups with respect to quartiles (≤25; >25 to ≤50; >50 to ≤75; and >75). No significant disease‐specific survival difference among the quartiles was found based on analysis of intensity (p=0.689) and extent (p=0.975) of COX‐2 staining. These results suggest that the expression of COX‐2 protein has no significant impact on the outcome of patients with colorectal cancer.