Математичні основи визначення функціонального стану операторів складних технологічних об’єктів

Забезпечення високої надійності роботи оператора, а, відповідно, системи «людина – машина» (СЛМ), – є пріоритетним завданням для більшості сфер промисловості та сучасного виробництва. Тому важливим є розробка заходів по підвищенню кваліфікації операторів складних технологічних об’єктів (СТО), шляхом підбору математичного апарату для який б дав можливість підвищити надійність його діяльності в структурі СЛМ

Immunohistochemische Lokalisation von Antikörpern gegen Bestandteile der photosensorischen Membran im Evertebraten-Photorezeptor

Eight biochemically characterized monoclonal mouse-anti-crayfish rhabdom antibodies (62D, 109, 46a, 16c, 151, 160, -Rh, 22A) and one polyclonal rabbit-anti-crayfish rhabdom antiserum (RAAS) were tested for their binding capacity to histologieal sections of Astacus retinae. The retinae were light- or dark-adapted and were fixed differently according to the method of marking (aldehyde fixation, cryo-fixation). All antibodies were investigated both by light and by electron microscopy. For the light microscopy a fluorescence and a gold-silver method were applied. Both methods yielded distinct rhabdomal markings for the RAAS. The monoclonal antibodies, almost all of which bind to the same protein (165 KDa) in irrmuno-bloc tests (with the exception of -Rh), react differently in the immuno-histochemical tests. The following results were obtainedby the light microscopical investigations: Four of the antibodies (46a, 16c, 151, 160) failed to bind to the sections in all experiments. Two IgG antibodies (62D, 109) showed distinct binding to the rhabdoms in the fluorescence tests and weak binding in the gold-silver markings. The preparation method (with or without the thiol-protease inhibitor Ep 475/tannic acid) as well as the state of adaptation (light/dark) did not influence these results. The monoclonal antibody -Rh directed against crayfish rhodopsin was tested only in gold-silver markings and was found to bind to the rhabdoms. With the IgM antibody (22A) no rhabdomal markings could be shown in light microscopical experiments. In gold-silver staining experiments, however, this antibody binds to the retinula Gell nuclei, if the retinae has not been previously: treated with Ep475 and/or tannic acid. [...

Filaggrin deficiency results in a dose-dependent impairment of the epidermal barrier in ichthyosis vulgaris

Oncogene-induced senescence acts as a barrier against tumor formation and has been implicated as the mechanism preventing the transformation of benign melanocytic lesions that frequently harbour oncogenic B-RAF or N-RAS mutations. In the present study we systematically assessed the relative importance of the tumor suppressor proteins p53, p21Waf1, pRb and p16INK4a in mediating oncogene-induced senescence in human melanocytes. We now show that oncogenic N-RAS induced senescence in melanocytes is associated with DNA damage, a potent DNA damage response and the activation of both the p16INK4a/pRb andp53/p21Waf1 tumor suppressor pathways. Surprisingly neither the pharmacological inhibition of the DNA damage response pathway nor silencing of p53 expression had any detectable impact on oncogene-induced senescence in human melanocytes. Our data indicate that the pRb pathway is the dominant effector of senescence in these cells, as its specific inactivation delays the onset of senescence and weakens oncogene-induced proliferative arrest.1 page(s

Interferon alpha signalling and its relevance for the upregulatory effect of transporter proteins associated with antigen processing (TAP) in patients with malignant melanoma

Introduction: Interferon alpha (IFNα) is routinely used in the clinical practice for adjuvant systemic melanoma therapy. Understanding the molecular mechanism of IFNα effects and prediction of response in the IFNα therapy regime allows initiation and continuation of IFNα treatment for responder and exclusion of non-responder to avoid therapy inefficacy and side-effects. The transporter protein associated with antigen processing-1 (TAP1) is part of the MHC class I peptide-loading complex, and important for antigen presentation in tumor and antigen presenting cells. In the context of personalized medicine, we address this potential biomarker TAP1 as a target of IFNα signalling. Results: We could show that IFNα upregulates TAP1 expression in peripheral blood mononuclear cells (PBMCs) of patients with malignant melanoma receiving adjuvant high-dose immunotherapy. IFNα also induced expression of TAP1 in mouse blood and tumor tissue and suppressed the formation of melanoma metastasis in an in vivo B16 tumor model. Besides its expression, TAP binding affinity and transport activity is induced by IFNα in human monocytic THP1 cells. Furthermore, our data revealed that IFNα clearly activates phosphorylation of STAT1 and STAT3 in THP1 and A375 melanoma cells. Inhibition of Janus kinases abrogates the IFNα-induced TAP1 expression. These results suggest that the JAK/STAT pathway is a crucial mediator for TAP1 expression elicited by IFNα treatment. Conclusion: We suppose that silencing of TAP1 expression provides tumor cells with a mechanism to escape cytotoxic T-lymphocyte recognition. The observed benefit of IFNα treatment could be mediated by the shown dual effect of TAP1 upregulation in antigen presenting cells on the one hand, and of TAP1 upregulation in ‘silent’ metastatic melanoma cells on the other hand. In conclusion, this work contributes to a better understanding of the mode of action of IFNα which is essential to identify markers to predict, assess and monitor therapeutic response of IFNα treatment in the future

IFNα-2b (IntronA) stimulates TAP1 expression in peripheral blood mononuclear cells (PBMC) of patients (n = 18) with malignant melanoma receiving adjuvant high-dose immunotherapy.

<p>(A) The actual administered dose of IFNα-2b was about 20 million IU/m² dependent on the clinically observed side effects. Only days of IFNα treatment are shown. (B) mRNA expression of TAP1 in PBMCs of patients treated with adjuvant high-dose IFNα-2b. Statistical analysis of TAP expression was performed using the <i>Statistical Analysis System</i> of the SAS Institute Inc. (Cary, NC, USA).</p

Stimulatory effects of IFNα on TAP expression, peptide binding and transport.

<p>THP1 cells and A375 cells were treated with the indicated concentrations of IFNα for 3 hours (A, B). After stimulation, TAP1 mRNA and 18SrRNA mRNA (as internal reference) were measured by qRT-PCR analysis. Depicted are -fold changes relative to untreated cells = standard error of mean (SEM) of three independent experiments. (C) TAP1 protein expression is induced by IFNα in THP1 cells measured by western blot analysis. (D) TAP dependent peptide-binding sites are increased significantly by IFNα in THP1 cells (p = 0.0072). (E) ATP-dependent peptide transport is stimulated significantly by IFNα in THP1 cells (p = 0.0006). Statistical analysis was performed using unpaired Student’s t-test (** p<0.01, *** p<0.001).</p

IFNα-activated signalling pathways in THP1 and A375 cells.

<p>THP1 (A) and A375 (B) cells were stimulated with the indicated concentrations of IFNα. As controls, THP-1 cells were additionally stimulated with IL-6/sIL-6R and LPS; A375 cells were stimulated with OSM or IL-1β. The phosphorylation levels of STAT1, STAT3, ERK1/2 and Akt were detected via Western blot analysis. Displayed are mean values of at least three independent experiments.</p