The expression of SFRP1, SFRP3, DVL1, and DVL2 proteins in testicular germ cell tumors

Germ cell tumors of the testis are a heterogeneous group of neoplasms that affect male adolescents and young adults. Wnt signaling pathway components have been shown to be actively involved in normal and malignant germ cell differentiation and progression. In this study, we aimed to explore the expression patterns of the secreted frizzled‐related protein (SFRP) and Disheveled protein family (DVL) in a subset of testicular germ cell tumors. Eighty‐five formalin‐fixed, paraffin‐embedded tissue samples of the primary germ cell tumors of the testis were stained against SFRP1, SFRP3, DVL1, and DVL2 proteins using immunohistochemistry. SFRP1 and SFRP3 exhibited lower expression in both seminomas and mixed/non‐seminomatous tumors, compared with atrophic/benign tissue (p < 0.001). SFRP3 expression was lower than SFRP1 expression within the seminoma group (p = 0.004), but not within the mixed/non‐seminomatous group (p = 0.409). The majority of the tested cases (27/28, 96%) exhibited low DVL1 protein expression (median 0%, range 0–90%). In contrast, 20 out of 22 tested cases (91%) exhibited strong expression of DVL2 protein (median 80%, range 0–100%). No significant difference in DVL1 and DVL2 protein expression was observed between seminomas and mixed/non‐seminomatous tumors (p = 0.68 and 0.29). The secreted frizzled‐related protein and disheveled protein family members appear to be actively involved in the pathogenesis of primary testicular germ cell tumors.     

Influence of hyperthermal regimes on experimental teratoma development in vitro

We screened for the impact of hyperthermal regimes varying in the cumulative equivalent minutes at 43°C (CEM43°C) and media composition on tumour development using an original teratoma in vitro model. Rat embryos (three germ layers) were microsurgically isolated and cultivated at the air‐liquid interface. During a two week period, ectodermal, mesodermal and endodermal derivatives developed within trilaminar teratomas. Controls were grown at 37°C. Overall growth was measured, and teratoma survival and differentiation were histologically assessed. Cell proliferation was stereologically quantified by the volume density of Proliferating Cell Nuclear Antigen. Hyperthermia of 42°C, applied for 15 minutes after plating (CEM43°C 3.75 minutes), diminished cell proliferation (P ? .0001) and enhanced differentiation of both myotubes (P ? .01) and cylindrical epithelium (P ? .05). Hyperthermia of 43°C applied each day for 30 minutes during the first week (CEM43°C 210 minutes) impaired overall growth (P ? .01) and diminished cell proliferation (P ? .0001). Long‐term hyperthermia of 40.5°C applied for two weeks (CEM43°C 630 minutes) significantly impaired survival (P ? .005). Long‐term hyperthermia of 40.5°C applied from the second day when differentiation of tissues begins (CEM43°C 585 minutes) impaired survival (P ? .0001), overall growth (P ? .01) and cartilage differentiation (P ? .05). No teratomas survived extreme regimes: 43°C for 24 hours (CEM43°C 1440 minutes), hyperthermia in the scant serum‐free medium (CEM43°C 630 minutes) or treatment with an anti‐HSP70 antibody before long‐term hyperthermia 40.5°C from the second day (CEM43°C 585 minutes). This in vitro research provided novel insights into the impact of hyperthermia on the development of experimental teratomas from their undifferentiated sources and are thus of potential interest for future therapeutic strategies in corresponding in vivo models.     

Preferential recognition of a microbial metabolite by human Vgamma2Vdelta2 T cells

Human Vgamma2Vdelta2 T cells are stimulated by prenyl pyrophosphates, such as isopentenyl pyrophosphate (IPP), and play important roles in mediating immunity against microbial pathogens and have potent anti-tumor activity. (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) has been identified as a metabolite in the 2-C-methyl-D-erythritol-4 phosphate (MEP) pathway for isoprenoid biosynthesis that is used by many bacteria and protozoan parasites. We find that HMBPP is the major Vgamma2Vdelta2 T-cell antigen for many bacteria, including Mycobacterium tuberculosis, Yersinia enterocolitica and Escherichia coli. HMBPP was a 30 000-fold more potent antigen than IPP. Using mutant bacteria, we show that bacterial antigen levels for Vgamma2Vdelta2 T cells are controlled by MEP pathway enzymes and find no evidence for the production of 3-formyl-1-butyl pyrophosphate. Moreover, HMBPP reactivity required only germ line-encoded Vgamma2Vdelta2 TCR elements and is present at birth. Importantly, we show that bacterial HMBPP levels correlated with their ability to expand Vgamma2Vdelta2 T cells in vivo upon engraftment into severe combined immunodeficiency-beige mice. Thus, the production of HMBPP by a microbial-specific isoprenoid pathway plays a major role in determining whether bacteria will stimulate Vgamma2Vdelta2 T cells in vivo. This preferential stimulation by a common microbial isoprenoid metabolite allows Vgamma2Vdelta2 T cells to respond to a broad array of pathogens using this pathway.     

CD105 expression as a measure of microvessel density in supraglottic laryngeal squamous cell carcinoma

The purpose of the present study was to examine the expression of CD105 among patients with supraglottic laryngeal cancer and to assess the clinical relevance of CD105-assessed MVD. A total of 40 patients with supraglottic squamous cell carcinomas (SCCs) were included in the present study. Surgical specimens were immunostained for CD105 and MVD was calculated at 400× magnification. The rounded mean value of the vessel count in four fields for each case was used as the final MVD value. The mean MVD value assessed by CD105 in considered supraglottic SCCs was 13.5 (SD = 3.97). High MVD was significantly correlated with advanced (III and IV) clinical stage (Mann–Whitney U test P = 0.01) and malignancy recurrence presence/absence (Mann–Whitney U test P = 0.023). Spearman’s rank correlation test showed significant correlation between high CD105-assessed MVD and pN+ category (rho = 0.337, P = 0.033), advanced Stage (III and IV) (rho = 0.402, P = 0.01) and developed locoregional recurrence (rho = 0.395, P = 0.012). The logistic regression showed that a high CD105+ MVD was the only independent marker of tumor recurrence (P = 0.029; odds ratio, 6.64; 95% CI, 1.218–36.152). The average MVD was significantly higher in patients with advanced TNM stage and in patients with locoregional recurrence of disease, suggesting that angiogenesis is closely related with clinical aggressiveness of tumor. CD105-assessed MVD in supraglottic laryngeal SSCs may identify patients at risk of recurrence of disease.     

Automutilation of the penis performed by the kitchen's knife

Automutilation of the penis performed by the kitchen's knife is described. Surgical techniques, the most important goals of therapy as well as psychiatric aspects of the automutilation are discussed.     

Mg2+ sensitizes KATP channels to inhibition by DIDS: dependence on the sulphonylurea receptor subunit

1. ATP-sensitive potassium channels (K(ATP) channels) consist of pore-forming Kir6.x subunits and of sulphonylurea receptors (SURs). In the absence of Mg(2+), the stilbene disulphonate, DIDS, irreversibly inhibits K(ATP) channels by binding to the Kir subunit. Here, the effects of Mg(2+) on the interaction of DIDS with recombinant K(ATP) channels were studied in electrophysiological and [(3)H]-glibenclamide binding experiments. 2. In inside-out macropatches, Mg(2+) (0.7 mM) increased the sensitivity of K(ATP) channels towards DIDS up to 70 fold (IC(50)=2.7 micro M for Kir6.2/SUR2B). Inhibition of current at DIDS concentrations > or =10 micro M was irreversible. 3. Mg(2+) sensitized the truncated Kir6.2Delta26 channel towards inhibition by DIDS only upon coexpression with a SUR subunit (SUR2B). The effect of Mg(2+) did not require the presence of nucleotides. 4. [(3)H]-glibenclamide binding to SUR2B(Y1206S), a mutant with improved affinity for glibenclamide, was inhibited by DIDS. The potency of inhibition was increased by Mg(2+) and by coexpression with Kir6.2. 5. In the presence of Mg(2+), DIDS inhibited binding of [(3)H]-glibenclamide to Kir6.2/SUR2B(Y1206S) with IC(50)=7.9 micro M by a non-competitive mechanism. Inhibition was fully reversible. 6. It is concluded that the binding site of DIDS on SUR that is sensed by glibenclamide does not mediate channel inhibition. Instead, Mg(2+) binding to SUR may allosterically increase the accessibility and/or reactivity of the DIDS site on Kir6.2. The fact that the Mg(2+) effect does not require the presence of nucleotides underlines the importance of this ion in modulating the properties of the K(ATP) channel.     

Effects of transient loss of shear stress on blood-brain barrier endothelium: role of nitric oxide and IL-6

Loss of blood-brain barrier (BBB) function may contribute to post-ischemic cerebral injury by yet unknown mechanisms. Ischemia is associated with anoxia, aglycemia and loss of flow (i.e. shearing forces). We tested the hypothesis that loss of shear stress alone does not acutely affect BBB function due to a protective cascade of mechanisms involving cytokines and nitric oxide (NO). To determine the relative contribution of shear stress on BBB integrity we used a dynamic in vitro BBB model based on co-culture of rat brain microvascular endothelial cells (RBMEC) and astrocytes. Trans-endothelial electrical resistance (TEER), IL-6 release and NO levels were measured from the lumenal and ablumenal compartments throughout the experiment. Flow-exposed RBMEC were challenged with 1 h of normoxic-normoglycemic flow cessation (NNFC) followed by reperfusion for 2 to 24 h. NNFC caused a progressive drop in nitric oxide production during flow cessation followed by a time-dependent increase in ablumenal IL-6 associated with a prolonged NO increase during reperfusion. The nitric oxide synthetase (NOS) inhibitor L-NAME (10 microM) abrogated all effects of NNFC, including changes in NO and cytokine production. BBB permeability did not increase during or after NNFC/reperfusion, but was increased by treatment with L-NAME or when the effects of IL-6 were blocked. Flow adapted RBMEC and astrocytes respond to NNFC/reperfusion by overproduction of IL-6, possibly secondary to increased production of NO during the reperfusion. Maintenance of BBB function during and following NNFC appears to depend on intact NO signaling and IL-6 release.     

Determination of benzene in urine by static headspace gas chromatography

This paper describes the application of static headspace gas chromatography in determining benzene in urine. The method was analytically validated for sensitivity (DL = 42 ng/l), repeatability (RSD = 3% and 4%), and accuracy (71%), and was applied in measuring urine benzene in nonsmokers (N = 14) and smokers (N = 18). All urine samples had measurable benzene concentrations. The method proved sensitive enough to establish a significant statistical difference (P < 0.000614) in urine benzene concentrations between smokers (mean = 760; range = 181-1869 ng/l) and nonsmokers (mean = 214; range = 61-515 ng/l).     

Evaluation of a community-level health policy intervention

In this article an evaluation approach towards an innovative community-level health policy intervention in Germany is presented. The aim of the intervention was basically to establish new structures of health planning and co-ordination at the community level in order to improve health monitoring and health care as well as health promotion. To realise this aim Round Tables, Working Groups and Project Offices were implemented in the communities. Based on the theory of change and using a developmental perspective the evaluation was focussed on the links between contexts, activities and outputs of the initiative. A mixed-method design with triangulation of quantitative and qualitative methods was applied. Results show that most of the communities succeeded in convening all major organisations and institutions of the local health care system within the Round Tables. Working climate was rated favourably by most of the involved actors. All communities succeeded in developing and enacting recommendations for action programmes, and about 40% of these programmes were implemented during the observation period of the project. Probability of programme implementation was high if measures remained within the scope of the communities' range of decision authority. Potential effects on population health produced by the action programmes could not be assessed both for logistic reasons and due to the short observation period. Finally, some major characteristics of health policy evaluation at the community level are discussed briefly.