Autologous and Pooled Tumor Lysates in Combined Immunotherapy of Patients with Glioblastoma.

Although major progress has been made in the standard treatment for glioblastomas, encompassing the maximal surgical resection, chemotherapy and radiation therapy, it is possible to increase survival rates significantly only in a few patients. Therefore, it is necessary to explore new therapeutic modalities, one of which is immunotherapy. The aim of the study was to evaluate the efficacy of the combined use of autologous and pooled tumor lysates in comprehensive treatment of patients with glioblastoma. MATERIALS AND METHODS: All patients (n=58, including 30 males and 28 females aged 18-70 years) were randomized into three groups, two of which received immunotherapy based on injection of autologous dendritic cells pulsed with autologous tumor lysates (first protocol) or pooled lysates (second protocol) from more than one tumor, in addition to the planned standard treatment. The patients of group 3 (control) received the standard comprehensive treatment encompassing the maximum safe tumor resection followed by radiation therapy and chemotherapy. RESULTS: The tolerability of both applied immunotherapy protocols was good: there were no anaphylactic reactions observed or patients who prematurely discontinued participation in the study. The final analysis of the data revealed no significant differences in median survival values of patients in each of the three groups. However, when analyzing the Karnofsky Performance Status in patients of group 2, it was found that it tended to improve. CONCLUSION: The study shows that the proposed immunotherapy protocols are safe for clinical use and have the potential to improve the patient's life quality. However, these findings should be considered intermediate until the findings of multicenter randomized clinical trials with a larger number of patients are obtained.

[THE HEALTH SAVING TECHNOLOGIES AT A PEDAGOGICAL UNIVERSITY].

The article investigates health saving technologies at a pedagogical university, theirs types and classifications, psychological and pedagogical aspects of application in education. It has proved that health saving technologies formed special knowledge, skills, skills to maintain and strengthen health, the formation of an individual healthy way of life, should provide the basis for independent attempts to improve himself / herself, his /her body, psyche, emotions. Introduced the fact that the purpose of the article was the scientific and theoretical substantiation of health saving educational technologies students studies at a pedagogical university, special attention was paid to color therapy, music therapy, fairy tale therapy, laugh therapy, etc. Analytical, diagnostic, experimental and systematic methods were used. The problem of the development, implementation and use in pedagogical activities of health saving educational technologies by students of pedagogical universities is a priority, because future teachers must ensure the intellectual, moral volitional, aesthetic, physical, emotional and moral development of the personality of the person.

[Developmental changes of neurotransmitter properties in sympathetic neurons].

Neurochemical composition of metasympathetic nervous system is characterized by a large variation. The main part of the intramural ganglionic neurons is cholinergic. Along with cholinergic neurons, there are ganglionic neurons containing serotonin, histamine, GABA, and several peptides: cholecystokinin, dynorphin, enkephalin, galanin, gastrin-releasing peptide (bombesin in mammals), neuropeptide Y, neurotensin, somatostatin, tachykinins, neurokinin A, vasoactive intestinal polypeptide and calcitonin gene related peptide. Gases as NO, CO, H2S, also act as neurotransmitters. Separate groups of neurons differ in the content of neuronal calcium-binding proteins, such as calbindin, calretinin and parvalbumin and neurofilaments: low molecular weight, a medium molecular weight and high molecular weight. Neurons of the enteric ganglia are the most different by their neurochemistry. There is a species difference in the ganglia of large animals and humans there are more combinations of chemical transmitters. Synthesis of neurotransmitters takes place even in the embryonic period and by the time of birth the most of neurons contain acetylcholine. In postnatal ontogenesis, the proportion of neurons expressing the NO-synthase decreases in the enteric and cardiac intramural ganglionic neurons. The functional significance of these changes is unclear.

Interleukin-1 receptors type I and type II are differentially regulated in human keratinocytes by ultraviolet B radiation.

Since regulation of keratinocyte IL-1 receptor expression is likely to have a major impact on the biologic effects of IL-1 on epidermal cells, we examined expression, regulation, and function of IL-1R in cultured human keratinocytes. By reverse transcriptase polymerase chain reaction, human keratinocytes were shown to express IL-1 receptor type I (IL-1RI) and IL-1 receptor type II (IL-1RII). Human keratinocyte IL-1RI mRNA expression was dependent on the differentiation state of the cell and was regulated by ultraviolet B (UVB) radiation, which initially decreased but later increased IL-1RI expression. This UVB-induced biphasic modulation of IL-1RI expression was mediated by an autocrine mechanism involving endogenously produced IL-1alpha and IL-1RI. Increased expression of IL-1RI in UVB-irradiated or IL-1alpha-stimulated keratinocytes was functionally important, because it endowed these cells with the capacity to upregulate expression of the intercellular adhesion molecule (ICAM)-1 upon IL-1alpha stimulation. Keratinocyte IL-1RII expression was regulated by UVB irradiation in an inverse manner. Significant and rapid upregulation of IL-1RII was observed within 1 h after UVB irradiation and gradually decreased to background levels within 24 h. Inverse regulation of IL-1RII versus IL-1RI was associated with opposite functions, because blocking of IL-1RII enhanced IL-1alpha effects on induction of ICAM-1 expression. These studies demonstrate that IL-1 responsiveness of UVB-irradiated keratinocytes critically depends on regulation of IL-1RI expression and that IL-1RII serves as a "decoy" receptor for IL-1, limiting rather than promoting IL-1-mediated effects.

Analysis of the production of soluble ICAM-1 molecules by human cells.

The adhesion molecule intercellular adhesion molecule-1 (ICAM-1), in addition to its membrane-associated form (mICAM-1), also exists as a soluble form (sICAM-1). sICAM-1 is capable of binding to lymphocyte function associated antigen-1 (LFA-1) molecules, and production of sICAM-1 is therefore thought to have immunomodulatory consequences. The present study, which employed normal human keratinocytes as a model for sICAM-1-producing cells, was conducted to determine the mechanism responsible for the production of sICAM-1 and to develop a strategy for specific inhibition of sICAM-1 production. Stimulation of keratinocytes with recombinant human gamma-interferon (rhIFN-gamma) induced both expression of mICAM-1 and production of sICAM-1. Western blot analysis revealed that keratinocyte-derived sICAM-1, compared to mICAM-1, had a smaller molecular size, approximately a 7-kD difference. Neither by Northern blot analysis nor by reverse-transcriptase polymerase chain reaction (RT-PCR) was any evidence for alternatively spliced ICAM-1 mRNA obtained. Addition of the protease inhibitors iodoacetamide and E-64, however, inhibited the production of sICAM-1 in a dose-dependent manner. The involvement of proteolytic cleavage in the production of sICAM-1 was corroborated in minimal peptide protection assays, in which minimal peptides covering the potential cleavage site of ICAM-1 were added to sICAM-1-producing keratinocytes. One of these peptides, ICAM cleavage inhibitory peptide (ICAM-CIP), inhibited the production of sICAM-1 without affecting mICAM-1 expression. These studies demonstrate that sICAM-1 production in human keratinocytes is due to proteolytic cleavage, and that the oligopeptide ICAM-CIP may specifically inhibit this mechanism. The capacity of ICAM-CIP to selectively prevent production of sICAM-1 may be useful for the development of novel therapeutic approaches relevant for the management of inflammation and cancer.

[Effect of various thawing techniques on the quality of small fruit frozen products]. / Wplyw róznych technik rozmrazania mrozonek z owoców jagodowych na ich jakosc.

The investigation concerned the effect of three methods of thawing frozen products at 18-20 degrees C (room temperature), 2-4 degrees C (conditions of a domestic refrigerator), and in a microwave oven, on the quality of fruit of blue berry, raspberry, black- and red currant, and strawberry. Physico-chemical indices applied as the criteria of the estimate were the content of dry matter, volatile acids, vitamin C, and anthocyanins, the amount of cell sap exuded after thawing, and the results of organoleptic evaluation. After harvest fruits were kept in a cold store and processed within 24 h. Frozen products were stored at about -30 degrees C up to the time of degustation. From the practical point of view the applied techniques of thawing insignificantly affected the level of the physico-chemical indices. Only in the case of fruit affected by microwaves the content of vitamin C (black currant) and anthocyanins (blue berry, raspberry, and black currant) was smaller, though maximum differences did not exceed 10%. The amount of cell sap exuded in the course of thawing depended on the fruit species and method of thawing. Blackcurrant exuded trace amount of sap, greater ones were noted, in the increasing order, with bilberry, raspberry, and strawberry, and the greatest with redcurrant. Of the thawing techniques tested the smallest amount of sap exuded at 2-4 degrees C, at 18-20 degrees it was by 30% and in the microwave oven by 63% greater.(ABSTRACT TRUNCATED AT 250 WORDS)

Post-transcriptional regulation of interleukin-6 gene expression in human keratinocytes by ultraviolet B radiation.

Exposure to increasing doses (290-315 nm) of ultraviolet (UV) B radiation is thought to profoundly affect human health. Studies on the biologic and molecular effects of UVB radiation on human skin are therefore of particular interest. There is experimental and clinical evidence to assume that UVB radiation-induced local and systemic inflammatory reactions might be mediated at least in part by UVB-induced keratinocyte-derived interleukin (IL)-6. Previously, a UVB-induced increase of steady-state levels of IL-6 mRNA was found to be a prerequisite for keratinocyte IL-6 production after UVB irradiation. The present study was aimed at addressing the question of whether in vitro UVB irradiation would increase IL-6 mRNA expression in long-term cultured, normal human keratinocytes via transcriptional or post-transcriptional mechanisms. UVB exposure (0-100 J/m2) of keratinocytes increased low baseline expression levels of IL-6 mRNA in a time- and dose-dependent manner. Using nuclear run-on assays, transcription rates of the IL-6 gene in nuclei isolated from UVB-irradiated cells were found to be essentially identical to those seen in unirradiated cells, indicating that UVB light did not lead to increased transcription of the IL-6 gene. To determine a possible post-transcriptional mechanism in UVB-induced IL-6 mRNA expression, the effects of UVB irradiation on IL-6 mRNA stability were examined. To this end irradiated and unirradiated keratinocytes were treated with actinomycin D and subjected to Northern blot analysis to calculate IL-6 mRNA half-life. As compared with unirradiated cells, IL-6 mRNA stability was increased significantly (three- to four-fold) in UVB-irradiated cells, suggesting that UVB radiation upregulates IL-6 mRNA levels in human keratinocytes by increasing the stability of IL-6 transcripts. This is the first report indicating that UVB radiation at a physiologically relevant dose may affect gene expression in human cells at a post-transcriptional level.

The 55-kD tumor necrosis factor receptor on human keratinocytes is regulated by tumor necrosis factor-alpha and by ultraviolet B radiation.

In previous studies we showed that cultured human keratinocytes expressed the 55-kD TNF receptor (TNFR) and that its expression the important for TNF alpha-mediated upregulation of intercellular adhesion molecule-1 (ICAM-1) expression on keratinocytes. Because factors that either reduce or enhance TNFR expression are likely to have a major impact on the biological effects of TNF alpha on keratinocytes, these studies were conducted to determine the factors that regulate its expression on keratinocytes. Using reverse transcriptase polymerase chain reaction, human keratinocytes were shown to lack 75-kD TNFR expression, indicating that TNF responsiveness of human keratinocytes critically depended on regulation of 55-kD TNFR expression. Human keratinocyte 55-kD TNFR surface and mRNA expression was found to be regulated in vitro by recombinant human (rh) TNF alpha. Stimulation of keratinocytes with rhTNF alpha initially decreased, but later increased, 55-kD TNFR surface expression. This biphasic modulation of 55-kD TNFR surface expression was associated with concomitant changes in 55-kD TNFR mRNA expression. Ultraviolet B (UVB) radiation, a well-known inducer of synthesis and secretion of TNF alpha by human keratinocytes, was found to mimic TNF alpha-induced modulation of 55-kD TNFR surface and mRNA expression via a TNF alpha-mediated autocrine regulatory mechanism. Production of soluble 55-kD TNFR by human keratinocytes remained unaffected by TNF alpha stimulation or UVB irradiation. These studies provide clear evidence that membrane expression of the human 55-kD TNFR may be regulated in human keratinocytes by the ligand itself: TNF alpha. Since in previous studies UVB irradiation transiently inhibited TNF alpha-induced human keratinocyte ICAM-1 expression, it is proposed that UVB radiation-induced biphasic modulation of human keratinocyte 55-kD TNFR expression may affect the capacity of these cells to respond to TNF alpha.

Induction of intercellular adhesion molecule 1 (ICAM-1) expression in normal human eosinophils by inflammatory cytokines.

Intercellular adhesion molecule-1 (ICAM-1) functions as a ligand for lymphocyte function-associated antigen-1 (LFA-1), and thereby plays a crucial role in mediating cell-cell interactions in inflammatory reactions. Human eosinophils represent important effector cells in allergic skin diseases. To gain more insight into the capacity of eosinophils to physically interact with LFA-1-positive inflammatory leukocytes, in the present study ICAM-1 expression in eosinophils was investigated. Using fluorescence-activated cell sorter analysis, it could be shown that highly purified (> or = 95%) eosinophils from peripheral blood of non-atopic individuals do not constitutively express ICAM-1 molecules. However, stimulation of eosinophils with interferon gamma (IFN gamma), tumor-necrosis factor alpha (TNF alpha), or interleukin 3 (IL-3) markedly upregulated ICAM-1 surface expression in a time- and dose-dependent manner. Cytokine-induced ICAM-1 expression in human eosinophils was corroborated by Northern blot analysis. Accordingly, unstimulated eosinophils did not express significant amounts of ICAM-1 mRNA, but ICAM-1 mRNA expression could be markedly induced in these cells upon stimulation with IFN gamma plus TNF alpha. The combination of TNF alpha with either IFN gamma, IL-3, IL-5, or granulocyte/macrophage colony-stimulating factor (GM-CSF) increased ICAM-1 expression in a synergistic fashion, whereas IL-5 or GM-CSF by itself did not induce ICAM-1 expression. Cytokine-induced ICAM-1 expression was specific, because IL-1 alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-7, IL-8, C5a, and platelet-activating factor did not significantly affect eosinophil ICAM-1 surface expression. In summary, these studies indicate that eosinophils may be activated to express the adhesion molecule ICAM-1 upon stimulation with selected inflammatory cytokines, which may allow adhesion-mediated cross-talk between eosinophils and LFA-1-positive cells. In addition, these data demonstrate for the first time a role for IL-3, IL-5, and GM-CSF in regulation of ICAM-1 expression in human cells.

Human epidermal keratinocytes are a source of soluble ICAM-1 molecules.

A soluble form of the usually membrane-bound adhesion molecule ICAM-1 was detected in supernatants derived from human epidermal keratinocytes. Specifically, supernatants harvested from long-term cultured normal human keratinocytes, or from the spontaneously immortalized keratinocyte cell line HaCaT, did not contain significant amounts of sICAM-1, but shedding of sICAM-1 was found to be markedly induced upon stimulation of keratinocytes with rh IFN gamma. In contrast, cells from the two epidermoid carcinoma cell lines, KB and A431, constitutively shed significant amounts of sICAM-1 even without cytokine stimulation, and sICAM-1 contents in supernatants harvested from these cells were further increased upon stimulation of cells with rh IFN gamma. These studies indicate, that in addition to peripheral blood mononuclear cells and human melanoma cells, human epidermal keratinocytes constitute an important cellular source of sICAM-1. By binding to leukocyte LFA-1 molecules, keratinocyte-derived sICAM-1 may influence inflammatory responses in the skin. In addition, constitutive shedding of sICAM-1 by transformed human keratinocytes may represent a possible mechanism by which neoplastic keratinocytes escape from cytotoxicity.

Structure and topography of the cochlear nucleus in bison.

Investigations were performed in 3 bisons of Bialowieza herd. Brain stems were fixed in 10% formaldehyde and cut transversely in sections of 15 microns. Sections were stained with hematoxylin and luxol fast blue. The cochlear nucleus in bison is divided into ventral and dorsal cellular bands. The ventral nucleus is longer and it is subdivided into dorsal and ventral parts. It is formed by round and multipolar cells. The dorsal cochlear nucleus shows laminar structure. Within the nucleus the ependymal, molecular and polymorphic zones may be distinguished. The cells are round, fusiform and multipolar.

[Treatment of patients with cancer of the colon]. / Lechenie bol'nykh rakom obodochnoi kishki.

Experience in the treatment of 317 patients with cancer of the colon is discussed. Surgery was conducted in 226 patients, 91 patients received combined treatment (operation and intravenous injection of radioactive colloidal gold). Postoperative mortality was 5.99%. The main cause of death was peritonitis (4.1%) due to incompetence of the anastomosis. Combined treatment of patients with stage III cancer of the colon improves somewhat the late-term results of management.