[Comparative characteristics of the incidence of facial boils in military personnel and civilians]. / Sravnitel'naya kharakteristika zabolevaemosti furunkulom chelyustno-litsevoi oblasti u voennosluzhashchikh i grazhdanskikh lits.

THE AIM OF THE STUDY: To study the frequency and structure of the incidence of facial furuncle in military personnel of St. Petersburg and Leningrad region and civilians. To conduct a comparative analysis of the nature of the course, duration and outcome of the disease in military personnel and civilians (according to case histories). MATERIAL AND METHODS: From 5744 case histories of patients with purulent-inflammatory processes of the maxillofacial region treated at the clinic of the Department of ChLH and Surgical Dentistry of the Kirov Medical University and St. Petersburg State Medical Institution «City Hospital No. 15¼ from 2017 to 2019, 201 case histories of patients with facial furuncle were isolated and analyzed. RESULTS: According to the results of a study conducted from 2017 to 2019 in the clinic of the Department of CHLH and surgical Dentistry, a facial furuncle was detected in 65 cases. In the general structure of purulent-inflammatory diseases of the maxillofacial region, it was 3.1%. In the department of ChLH GB No. 15, 136 people were treated, which amounted to 3.7%. A lower percentage of the disease in military personnel is associated with early detection and timely initiation of treatment of purulent-inflammatory diseases of the skin. Weekly inspections of military personnel in military units, allow to avoid the development of complicated forms of their course. 95.0% of military personnel (47.0% of them contractors) and 51.0% of civilians were in the age group from 17 to 21 years. The main cause of the disease - hypothermia - in military personnel was 32.1%, in civilians - 20.9%. More often, the furuncle disease occurred in the spring: in military personnel in 44.6%, in civilians in 30.7% of cases. Localization of a boil in the buccal region occurred in military personnel in 15.4% of cases, and in civilians - in 20.8% of cases. CONCLUSION: To prevent the disease of facial frunculus in military personnel, it is necessary to improve methods of prevention, diagnosis and treatment of patients, to provide consultations of related specialists: dermatovenerologist, endocrinologist, immunologist. In addition, it is necessary to observe the rules of personal hygiene with the use of individual skin care products (according to the type of facial skin), hardening of the body and vitamin therapy.

[Immunogenetic predictors of a rapid virologic response to antiviral therapy in patients with chronic hepatitis C].

AIM: To determine the correlation between interleukin 28B (IL28B) gene polymorphism in patients with chronic hepatitis C (CHC), the presence or absence a rapid virologic response to antiviral therapy, and a number of immunological characteristics as a basis for a personalized approach to treating the patients. SUBJECTS AND METHODS: Seventeen CHC patients infected with hepatitis C virus genotype 1b were examined and underwent genetic testing for IL28B gene polymorphism for rs12979860 (CC, CT or TT genotypes) and rs8099917 (TT, TG or GG genotypes) using the modified method of adjacent samples, which revealed single nucleotide substitutions in the genes. Their immunological parameters were identified by a flow cytometry technique by taking into account whether a rapid virologic response had been achieved. RESULTS: The key phenomena of a rapid virologic response in the representatives of different IL28B genotypes are the nonspecific proliferative activity of blood natural killer cells before treatment, as well as the count of regulatory T cells before and 4 weeks after therapy start. CONCLUSION: To predict the efficiency of antiviral therapy for CHC, it is desirable to supplement genetic studies with immunological data.

Expression and prognostic value of transcription factor PROX1 in colorectal cancer.

BACKGROUND: PROX1 is a specific target of the ß-catenin/TCF pathway in the intestinal epithelium. It acts as a regulator of progression from a benign to a highly dysplastic phenotype in colorectal tumours. However, the clinical significance of PROX1 expression is not known. METHODS: We studied the prognostic value of immunohistochemical expression of PROX1 in a series of 517 patients with colorectal cancer (CRC). RESULTS: The majority of the tumour samples expressed PROX1 (91%, 471 out of 517). High PROX1 expression was associated with a poor grade of tumour differentiation (P<0.0001). In the subgroup of patients with colon cancer, high PROX1 expression was associated with unfavourable colorectal cancer-specific survival (CCSS) as compared with low PROX1 expression (CCSS 47% vs 62%; P=0.045; RR 1.47). The association between high PROX1 and poor outcome was further strengthened in female colon cancer patients (CCSS 38% vs 63%; P=0.007; RR 2.02). Nonetheless, in multivariate survival analysis PROX1 expression was not retained as an independent prognostic factor. CONCLUSION: High PROX1 expression is associated with a poor grade of tumour differentiation, and, in colon cancer patients, also with less favourable patient outcome. Our results strengthen the previous preclinical observations that PROX1 has a role in tumour progression in CRC.

[The etiological pattern of diseases in pregnant women with enhanced blood ALT and AST activities, admitted to the obstetric unit of infectious disease hospital].

AIM: to define a role of hepatotropic (HAV, HBV, HCV, and HDV) and opportunistic hepatotropic (HGV, CMV, EBV, HHV types 1, 2, and 6) viruses in the etiological pattern of diseases accompanied by enhanced blood AlAT and AsA T activities in pregnant women. SUBJECTS AND METHODS: Two hundred and eleven pregnant women, including 123 patients with chronic viral hepatitis, 74 with enhanced blood AlAT activity and no markers of viral hepatitis (EAlA T-NMVH), and 14 with acute viral hepatitis were examined. RESULTS: Most pregnant women with chronic HBV and HCV infections were found to have HBV DNA and HCV RNA in the blood in the presence of normal and enhanced activities of transaminases. In the EAlAT-NMVH group, there was none of the opportunistic hepatotropic viruses in more than 7% of cases. No genetic material of HAV, HBV, HCV, HDV, HGV, CMV, EBV, HHV types 1, 2, and 6 was found in the blood of all 10 patients with hepatitis of unspecified etiology. CONCLUSION: In the absence of serologic data supporting the presence of infectious pathology, blood testing using the polymerase chain reaction is of low informative value in detecting opportunistic hepatotropic viruses in pregnant women with hepatitis of unspecified etiology. However, by keeping in mind that the spectrum of opportunistic hepatotropic viruses is not confined to those included in this study, it is expedient to examine additionally pregnant women with enhanced blood AlAT and AsAT activity in order to identify TTV, B19V, HHV-8, SEN and NV-F in the blood.

[The study of parameters of mesenchymal stromal cells differentiation in donors and patients with aplastic anemia].

AIM: To examine ability of mesenchymal stromal cells (MSC) of the bone marrow (BM) for differentiation in adipogenic and osteogenic differentiation in donors and patients with aplastic anemia (AA). MATERIAL AND METHODS: We obtained MSC cultures from BM cells of donors and AA patients and induced differentiation of mesenchymal cells with use of relevant reagents. Morphological changes in MSC were studied with light microscopy. A relative level of expression of differentiation marker genes in MSC cultures before and after induction of differentiation was analysed with reverse transcription-polymerase chain reaction. RESULTS: By morphological characteristics, MSC cultures in AA patients before and after differentiation induction do not differ from donor cultures, but relative expression of the genes of differentiation markers demonstrated that expression was different in male and female donors; MSC before and after induction of differentiation differ in donors and AA patients. CONCLUSION: Further studies are needed for detection of functional changes in precursors of stromal microenvironment and understanding of the disease pathogenesis.

[Acute hepatitis of unspecified etiology: etiological structure and clinical-laboratory characteristics].

The straight line nucleic acids detection method of viruses and wide spectrum of virus antigens immunodiagnostics in acute hepatitis of unknown etiology patients has allowed verifying the diagnosis at 19% cases (a viral hepatitis A, C or E). Results of research do not allow to consider hepatotropic viruses HGV, TTV, PV B19, EBV, CMV, HHV 1, 2, 6 and 8 type, NV-F as etiological agents at the majority of patients of investigated group, and the data of the anamnesis and a clinical and laboratory picture of a current of disease does not allow to exclude at 29.4% of patients a drug-induced hepatitis. Despite detailed molecular-biological and immunological inspection of patients, at 37.9% of acute hepatitis of unknown etiology patients it was not possible to establish a connection with hepatitis and defined etiological factor (the infectious agent).

[Impaired expression of genes regulating homeostasis of stem hemopoietic cells in stromal cells of patients with aplastic anemia].

AIM: To evaluate expression of genes participating in regulation of hemopoietic stem cells (HSC) in the cells of stromal sublayer of bone marrow long-term cultures in patients with aplastic anemia (AA); to determine effects of parathyroid hormone (PTH) on stromal microenvironment and on its ability to maintain HSC homeostasis. MATERIAL AND METHODS: Gene expression in the sublayer of the adherent cells (SAC) was examined with RT-PCt. SAC was for a long time treated with PTH, then their ability to secure survival of early hemopoietic precursors was tested. Changes in the function of stromal cells and expression of some genes were compared in 9 AA patients and 14 donors. RESULTS: Stromal sublayer of AA patients is characterized by low expression of Ang-1 and VCAM-1 genes and high VEGF expression compared to mean level of healthy donors. PTH stimulates expression of different genes participating in HSC regulation in stromal cells of some patients and improves survival of early hemopoietic hemopoietic precursors on such sublayers. CONCLUSION: AA patients have severe defects in SAC interaction with stroma. In some cases the defects can be partially compensated with application of PTH.

Vascular endothelial growth factor receptors in the regulation of angiogenesis and lymphangiogenesis.

VEGFR-1 (Flt-1), VEGFR-2 (KDR) and VEGFR-3 (Flt4) are endothelial specific receptor tyrosine kinases, regulated by members of the vascular endothelial growth factor family. VEGFRs are indispensable for embryonic vascular development, and are involved in the regulation of many aspects of physiological and pathological angiogenesis. VEGF-C and VEGF-D, as ligands for VEGFR-3 are also capable of stimulating lymphangiogenesis and at least VEGF-C can enhance lymphatic metastasis. Recent studies have shown that missense mutations within the VEGFR-3 tyrosine kinase domain are associated with human hereditary lymphedema, suggesting an important role for this receptor in the development of the lymphatic vasculature.

Shear stress-induced atherosclerotic plaque composition in ApoE(-/-) mice is modulated by connexin37.

OBJECTIVE: Shear stress patterns influence atherogenesis and plaque stability; low laminar shear stress (LLSS) promotes unstable plaques whereas oscillatory shear stress (OSS) induces more stable plaques. Endothelial connexin37 (Cx37) expression is also regulated by shear stress, which may contribute to localization of atherosclerotic disease. Moreover, Cx37 reduces initiation of atherosclerosis by inhibiting monocyte adhesion. The present work investigates the effect of Cx37 on the phenotype of plaques induced by LLSS or OSS. METHODS: Shear stress-modifying casts were placed around the common carotid artery of ApoE(-/-) or ApoE(-/-)Cx37(-/-) mice, and animals were placed on a high-cholesterol diet for 6 or 9 weeks. Atherosclerotic plaque size and composition were assessed by immunohistochemistry. RESULTS: Plaque size in response to OSS was increased in ApoE(-/-)Cx37(-/-) mice compared to ApoE(-/-) animals. Most plaques contained high lipid and macrophage content and a low amount of collagen. In ApoE(-/-) mice, macrophages were more prominent in LLSS than OSS plaques. This difference was reversed in ApoE(-/-)Cx37(-/-) animals, with a predominance of macrophages in OSS plaques. The increase in macrophage content in ApoE(-/-)Cx37(-/-) OSS plaques was mainly due to increased accumulation of M1 and Mox macrophage subtypes. Cx37 expression in macrophages did not affect their proliferation or their polarization in vitro. CONCLUSION: Cx37 deletion increased the size of atherosclerotic lesions in OSS regions and abrogated the development of a stable plaque phenotype under OSS in ApoE(-/-) mice. Hence, local hemodynamic factors may modify the risk for adverse atherosclerotic disease outcomes associated to a polymorphism in the human Cx37 gene.

Folding units in calcium vector protein of amphioxus: Structural and functional properties of its amino- and carboxy-terminal halves.

Muscle of amphioxus contains large amounts of a four EF-hand Ca2+-binding protein, CaVP, and its target, CaVPT. To study the domain structure of CaVP and assess the structurally important determinants for its interaction with CaVPT, we expressed CaVP and its amino (N-CaVP) and carboxy-terminal halves (C-CaVP). The interactive properties of recombinant and wild-type CaVP are very similar, despite three post-translational modifications in the wild-type protein. N-CaVP does not bind Ca2+, shows a well-formed hydrophobic core, and melts at 44 degrees C. C-CaVP binds two Ca2+ with intrinsic dissociation constants of 0.22 and 140 microM (i.e., very similar to the entire CaVP). The metal-free domain in CaVP and C-CaVP shows no distinct melting transition, whereas its 1Ca2+ and 2Ca2+) forms melt in the 111 degrees -123 degrees C range, suggesting that C-CaVP and the carboxy- domain of CaVP are natively unfolded in the metal-free state and progressively gain structure upon binding of 1Ca2+ and 2Ca2+. Thermal denaturation studies provide evidence for interdomain interaction: the apo, 1Ca2+ and 2Ca2+ states of the carboxy-domain destabilize to different degrees the amino-domain. Only C-CaVP forms a Ca2+-dependent 1:1 complex with CaVPT. Our results suggest that the carboxy-terminal domain of CaVP interacts with CaVPT and that the amino-terminal lobe modulates this interaction.

Modulation of glial activation by astrocyte-derived protein S100B: differential responses of astrocyte and microglial cultures.

The astrocyte-derived protein S100B stimulates production of inducible nitric oxide synthase and nitric oxide (NO) in astrocytes [Hu et al., 1996, J. Biol. Chem. 271:2543], but its effect on microglia is not known. In addition, S100B's ability to modulate the activity of other glial activating agents has not been defined. In this study, we compared the ability of S100B to stimulate NO in cultures of rat primary astrocytes and the BV-2 murine microglial cell line, and investigated the effect of the combined action of S100B and other stimuli known to activate glial cells. S100B itself stimulated the production of NO in astrocytes, and did not modify or potentiated only weakly the NO production induced by interleukin-1 beta, tumor necrosis factor alpha, dibutyryl cyclic AMP, zymosan A or lipid A. In contrast, S100B alone did not induce NO in BV-2 cells but strongly potentiated NO production in the presence of lipid A but not zymosan A. The deletion of eight C-terminal amino acid residues in S100B leads to a loss of the effect of S100B on microglia but not on astrocytes. These results demonstrate that responses of glial cells to extracellular S100B can vary depending on the cell type, and suggest that different structural features of S100B are important for the protein's effects on microglia and astrocytes.

Cyclopentenone prostaglandin 15-deoxy-Delta(12,14)-prostaglandin J(2) acts as a general inhibitor of inflammatory responses in activated BV-2 microglial cells.

15-deoxy-Delta(12,14)-PGJ(2), a cyclopentenone derivative of PGD(2), was recently reported [Petrova et al., Proc. Natl. Acad. Sci. USA 96 (1999) 4668-4673] to suppress inducible nitric oxide synthase (iNOS) production in microglia and mixed glial cultures stimulated with lipopolysaccharide (LPS). We report here that in addition to suppressing iNOS production, 15d-PGJ(2) also decreases the production of tumor necrosis factor alpha (TNFalpha), interleukin-1 beta (IL-1beta) and cyclooxygenase-2 (COX-2) in LPS-stimulated BV-2 microglial cells, thereby acting as a general inhibitor of microglial activation. Concomitantly, 15d-PGJ(2) itself up-regulates the production of the antioxidant enzyme heme oxygenase-1 (HO-1) and increases intracellular total glutathione levels. To test if increased HO-1 levels were involved in the ability of 15d-PGJ(2) to block microglial activation, we used a HO-1 inhibitor that could block the activity of HO-1. The presence of the HO-1 inhibitor did not alter the 15d-PGJ(2)-induced inhibition of LPS-stimulated iNOS and TNFalpha protein levels, and led to only a partial reduction in the protection offered by 15d-PGJ(2) against LPS-induced nitrite production. These results suggest that HO-1 upregulation by 15d-PGJ(2) is not the primary pathway responsible for the anti-inflammatory action of 15d-PGJ(2) in microglial cells.

Screening in a cell-based assay for inhibitors of microglial nitric oxide production reveals calmodulin-regulated protein kinases as potential drug discovery targets.

A high-throughput screening (HTS) assay for inhibitors of nitric oxide (NO) production by activated microglia was developed and used to compare the relative activities of various anti-inflammatory compounds and cell-permeable protein kinase inhibitors. BV-2 cells, an immortalized line that retains phenotypic features of microglia and produces NO in response to lipopolysaccharide (LPS), were used in the activation paradigm for the HTS assay. A characteristic feature of the compounds that were the most potent dose-dependent inhibitors of NO production is their ability to modulate serine/threonine protein kinases. The anti-inflammatory compound K252a, an inhibitor of calmodulin (CaM)-regulated protein kinases, had one of the highest potencies in the assay. Other classes of kinase inhibitors, including the protein kinase A inhibitor H-89, the mitogen activated protein kinase inhibitors PD98059 and SB203580, and the tyrosine kinase inhibitor genistein, were less potent and efficacious than K252a or the general serine/threonine/tyrosine kinase inhibitor staurosporine. K252a suppresses production of the inducible nitric-oxide synthase (iNOS). The inhibitory effect of K252a is not due to cell toxicity and does not correlate with inhibition of NFkappaB nuclear translocation. The mechanism of action appears to involve inhibition of phosphorylation of the transcription factor CREB, a protein whose activity is modulated by phosphorylation by CaM-dependent protein kinases. These data suggest that signal transduction pathways mediated by CaM-dependent protein kinases warrant future study as potential drug discovery targets.

Ortho-effects of carboxyl, sulpho and arsono groups on the protonation and dissociation constants of mono-azo and bis-azo chromotropic acid derivatives.

The effect of carboxyl sulpho and arsono groups on the protonation and dissociation constants of mono-azo and bis-azo chromotropic acid derivatives has been investigated. From the value obtained for the effect of the substituents the equation; ortho-effect = log K(ortho)-log K para has been derived. The causes of the effect are discussed.

Arsenazo III and its analogues-VII Colour reactions of the rare earth elements with a new reagent-carboxynitrazo.

A new reagent, Carboxynitrazo, gives a colour reaction with some rare earth elements. For the individual rare earth elements the sensitivities of the reactions are very different. The difference is greatest for the elements of the Ce- and Y-subgroups. For the first subgroup the molar absorptivities are about 16 x 10(4) l.mole(-1) cm(-1), whereas most of the elements of the Y-group do not give any colour reaction. A possible procedure for the determination of lanthanum in the presence of ytterbium is discussed.

[Production of a SCAR marker in pea Pisum sativum L. using RAPD analysis]. / Sozdanie SCAR-markera u gorokha (Pisum sativum L.) na osnovanii RAPD-analiza.

A polymorphic 750-bp fragment, RAPD marker, specific to particular pea genotypes (line L-111 and the Nord cultivar) was identified. Using this RAPD marker, SCAR was obtained. SCAR inheritance in the first and second generations was studied and its dominant character was shown.

[Phenylarsine oxide, a tyrosine phosphatase inhibitor, induces an increase in the intracellular concentration of calcium in rat peritoneal macrophages and human fibroblasts]. / Ingibitor tirozinfosfataz fenilarzinoksid indutsiruet uvelichenie vnutrikletochnoi kontsentratsii Ca2+ v peritoneal'nykh makrofagakh krysy i fibroblastakh cheloveka.

Using Fura-2 microfluorimetry, phenylarsine oxide (PAO) (10-50 microM), a potent tyrosine phosphatase inhibitor, was shown to induce a dose-dependent increase in the free Ca2+ intracellular concentration in rat peritoneal macrophages and human foreskin fibroblasts. The PAO-induced increase in [Ca2+]i is not due presumably to depletion of intracellular Ca2+ stores but to mainly a stimulation of Ca2+ entry from the extracellular medium. This PAO-activated Ca2+ entry is attenuated by the following pharmacological agents. Organic and inorganic Ca2+ channel blockers: (nifedipine, verapamil and Ni2+); nonselective cation channel blocker niflumic acid; tyrosine kinase inhibitors genistein and methyl-2,5-dihydroxycinnamate; SH-reagents dithiothreitol parachloromercuribenzoate and N-ethylmaleimide; arachidonic acid metabolism inhibitors 4-bromophenacyl bromide, indomethacin and caffeic acid; microtubule disrupters vinblastine, colchicine and colcemide. On the contrary, microfilament disrupters, cytochalasin B and phalloidin, enhance PAO-activated Ca2+ entry. Our data suggest that the dynamic balance between tyrosine kinase and phosphatase activity may play a central role in the maintenance of homeostatic levels of [Ca2+]i both in unstimulated cells and after agonist application.

[Organic and inorganic blockers of potential-dependent Ca2+ channels inhibit store-dependent entry of Ca2+ into rat peritoneal macrophages]. / Organicheskie i neorganicheskie blokatory potentsialzavisimykh Ca2+- kanalov ingibiruiut depozavisimyi vkhod Ca2+ v peritoneal'nye makrofagi krysy.

The effect of organic and inorganic blockers of voltage-dependent Ca(2+)-channels on thapsigargin- and UTP-induced store-operated Ca(2+)-entry in Fura-2-loaded rat peritoneal macrophages was investigated. This store-dependent or "capacitative" Ca2+ influx stimulated by emptying the intracellular Ca(2+)-stores with endoplasmic Ca(2+)-ATPase inhibitor thapsigargin (0.5 microM) or purinergic agonist UTP (200 microM) is inhibited by the following pharmacological agents: two structurally distinct organic Ca(2+)-channel blockers nifedipine and verapamil; inorganic Ca(2+)-channel inhibitors Ni2+, La3+, Gd3+; nonselective cation channel blocker niflumic acid. Our data suggest that store-operated Ca2+influx channels of rat peritoneal macrophages share pharmacologic properties with L-type Ca(2+)-channels. Similar to trp-channels of Drosophila, they may resemble L-type Ca(2+)-channels lacking a voltage sensor.

[The creation of an SV40 virus-based vector as an experimental model for the study of gene expression]. / Sozdanie na osnove virusa SV40 vektora kak éksperimental'noi modeli dlia izucheniia ékspressii genov.

The purpose of the studies is to design the recombinant virus SV 40 where the C-end of the basic structural protein of virus P-1 was replaced by a synthetic sequence of the neuropeptide bradykinin. The recombinant virus SV (SV 40/Brd) was obtained. In this virus 60 n.p. with 3'-end of VP-1 gene was substituted for 36 n.p. synthetic gene of bradykinin without impairing the frame of translation. The biological activity of SV 40 (Brd) was tested on the cultured cells CV-1 permissive for this virus. An immunofluorescence method was used to detect T antigen and to examine the cytopathic action of this recombinant. The gene engineering design does not make the recombinant loose its biological properties typical of wild virus.

[Frequency of genes speA, speB, and speC in Streptococcus pyogenes strains and the identification of the infective agent by polymerase chain reaction]. / Chastota vstrechaemosti genov speABC v shtammakh Streptococcus pyogenes i identifikatsia vosbuditelia s pomoshch'iu PTSR.

In cultures of S. pyogenes isolated from patients and carriers in different territories of the Russian Federation the genes of erythorogenic toxins A, B and C (speA, speB and specC) were detected. The possibility of the identification of S. pyogenes by means of PCR on the basis of primers to erythrogenic toxin B was determined. Gene speB was detected in all S. pyogenes cultures under study and proved to be species specific. Genes speA and speC were detected, respectively, in 29.4% and 9.35% of the S. pyogenes cultures under study. A test system for the identification of S. pyogenes on the basis of primers to gene speB was developed. The prospects for the detection of genes speA and speC for intraspecific typing of this infective agent were evaluated.