RESUMEN
It is known that exogenous gangliosides (GL) inhibit acute inflammatory signals in different cells induced by Escherichia coli lipopolysaccharide (LPS). Until now the mechanisms underlying their effect are unknown. We hypothesize that the anti-inflammatory effect of GL is caused by their ability to modify TLR4 translocation into the lipid rafts. To test this hypothesis, we studied the effect of exogenous GL on LPS-induced inflammatory reactions associated with increased nitric oxide and prostaglandin E2 (PGE2) production in epithelial cells isolated from the frog Rana temporia urinary bladder. It was shown that preincubation of cells with GM1 and GD1a in the concentration range from 100 nm to 50 µM reduced the effect of 25 µg/ml LPS E. coli on the increase of NO and PGE2 production. The effect of LPS was also eliminated in the presence of polymyxin B, capable to interact with lipid A in LPS molecule, which makes it inaccessible for binding to TLR4. The subcellular fractionation of epithelial cells in the sucrose density gradient in combination with immunoblotting revealed that LPS stimulates translocation of TLR4 into the lipid rafts in the cytoplasmic membrane. Preincubation of cells with GM1 or GD1a at concentration 20 µM completely eliminated the effect of LPS. A similar effect was revealed with 1 mM methyl-ß-cyclodextrin, a classical destructor of the lipid rafts. The results indicate the existence of a previously unknown mechanism of the anti-inflammatory effect of exogenous GL associated with their ability to interfere with LPS-induced translocation of TLR4 into the lipid rafts preventing LPS signal transduction. It is assumed that the observed effect of GL is based on their incorporation into cytoplasmic membrane and modification of the lipid rafts organization.
Asunto(s)
Células Epiteliales/metabolismo , Gangliósido G(M1)/farmacología , Gangliósidos/farmacología , Lipopolisacáridos/toxicidad , Animales , Células Cultivadas , Células Epiteliales/patología , Inflamación/inducido químicamente , Inflamación/metabolismo , Inflamación/patología , Rana temporariaRESUMEN
The effect of bacterial lipopolysaccharide (LPS) from E. coli on the viability of PC12 neuronal cell line was studied. LPS of 0111:B4 serotype and LPS of 055:B5 serotype were shown to have toxic effect on PC12 cells. But the toxic effect of LPS of 0127:B8 serotype was not revealed. Preincubation of PC12 cells with GM1 or GD1a gangliosides was found to diminish significantly the death of the cells and the formation of reactive oxygen species induced by LPS of 0111:B4 and 055:B5 serotypes in them. The protective effect of GM1 and GD1a was found not to depend on the presence of the inhibitor of Trk receptor tyrosine kinase in the medium, though activation of this protein kinase was previously shown to mediate the protective effect of gangliosides against the action of excitatory amino acids, pro-oxidants and other toxins on neurons and cells of neuronal cell lines. The protective effect of gangliosides against the LPS action on PC12 cells was similar to the effect of methyl-beta-cyclodextrin, which was shown to disturb cell membrane raft structure. The suggestion is put forward that the pronounced diminution of toxic effect of LPS on PC12 cells by gangliosides may be explained by the alteration of structural organization of lipid rafts caused by the incorporation of exogenous gangliosides in cell plasma membranes, which diminishes the translocation of TLR4 receptor into the rafts and their activation by LPS.
Asunto(s)
Supervivencia Celular/efectos de los fármacos , Gangliósidos/administración & dosificación , Lipopolisacáridos/toxicidad , Neuronas/efectos de los fármacos , Animales , Escherichia coli/química , Peróxido de Hidrógeno/metabolismo , Células PC12 , Ratas , Especies Reactivas de Oxígeno/metabolismo , Serogrupo , Transducción de Señal/efectos de los fármacos , Receptor Toll-Like 4/metabolismoRESUMEN
Earlier we have shown that in epithelial cells of the frog urinary bladder under action of bacterial lipopolysaccharides (LPS) there is activated expression of inducible NO-synthase (iNOS) and there is increased the NO production, which can play an important role in providing protective cell reactions from pathogens. The goal of the present work consisted in study of cyclooxigenase (cOG) products and mechanisms of their regulatory effect on expression of iNOS under action of LPS. In experiments on urinary bladder epithelial cells on the frog Rana temporaria it has been shown that incubation of the cells for 21 h with LPS leads to a rise in production of PGE2 and nitrites, stable NO metabolites. Inhibitor of iNOS 1400W decreased sharply production of nitrites, but did not affect the PGE2 level. Both the basal and the LPS-stimulated level of PGE2 and nitrites were inhibited in the presence of selective cOG inhibitors--SC-560 (cOG-1) and NS-398 (cOG-2). The IC50 value amounted to 90, 220, and 470 microM for NS-398, SC-560, and diclofenac (unspecific inhibitor of both isoforms), respectively. PGE2 and butaprost, the EP2-receptor agonist, but not agonists of EP1/EP3 or EP1 receptors, partially eliminated the inhibitory action of diclofenac on production of nitrites. Action of PGE2 was accompanied by an increase in the intracellular cAMP. Analysis of expression of iNOS mRNA in the epithelial cells incubated with LPS or LPS + inhibitor of cOG has shown the LPS-stimulated rise in expression of iNOS mRNA to decrease sharply in the presence of SC-560 or NS-398. Thus, the epithelial cells of the frog urinary bladder have the effectively functioning system of the congenital immune protection against bacterial pathogens, the most important component of this system being PGE2 and NO. Analysis of mechanisms of regulatory interactions of cOG and iNOS indicates that in this cell type the main regulators of iNOS expression and of the nitrogen oxide level are products of the cOG catalytic activity.
Asunto(s)
Óxido Nítrico Sintasa de Tipo II/metabolismo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Rana temporaria/inmunología , Vejiga Urinaria/inmunología , Urotelio/inmunología , Animales , Células Cultivadas , Inhibidores de la Ciclooxigenasa/farmacología , Diclofenaco/farmacología , Dinoprostona/química , Regulación de la Expresión Génica , Óxido Nítrico Sintasa de Tipo II/genética , Nitritos/metabolismo , Nitrobencenos/farmacología , Óxidos de Nitrógeno/química , Polisacáridos Bacterianos/inmunología , Prostaglandina-Endoperóxido Sintasas/química , Prostaglandina-Endoperóxido Sintasas/genética , Pirazoles/farmacología , Sulfonamidas/farmacología , Vejiga Urinaria/enzimología , Urotelio/enzimologíaRESUMEN
Since Gram-negative bacteria are known to be present in the cavity of urinary bladders in amphibian species, it was interesting to study the effect of bacterial endotoxins on epithelial signaling network which provides the arginine-vasotocin-induced increase of osmotic water permeability (OWP). The effect of LPS E. coli on AVT-induced OWP was studied in isolated frog Rana temporaria L. urinary bladder incubated during 20-21 hours in modified L-15 culture medium in sterile conditions. The LPS (25 microg/ml) was added into the mucosal solution. It was shown that exposure to LPS caused a strong suppression of the increase of OWP under AVT (0.5 nM), forskolin (35 microM) or IBMX (200 microM). Moreover, LPS induced more than 2-folds decrease both ofbasal and AVT-stimulated content of cAMP in the bladder tissue. The inhibitory effect of LPS on AVT-induced increase of OWP was eliminated in the presence of ODQ, 20 microM, a cytosolic guanylate cyclase inhibitor. With the use of RT-PCR it was shown that the expression of mRNA iNOS was 10-fold increased in 6 hours after LPS administration. These findings demonstrate the ability of frog bladder mucosal epithelial cells to recognize bacterial LPS and initiate antipathogen immune response related to increased production of nitric oxide. The activation of signal transduction cascade mediated by the LPS-induced immune response leads to a decrease of intracellular cAMP and down-regulates AVT-stimulated OWP acting at least in part through NO/cGMP-dependent signaling pathway.
Asunto(s)
Escherichia coli/inmunología , Lipopolisacáridos/inmunología , Ósmosis , Vejiga Urinaria/microbiología , Vejiga Urinaria/fisiología , Animales , AMP Cíclico/análisis , GMP Cíclico/análisis , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Lipopolisacáridos/farmacología , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Ósmosis/efectos de los fármacos , Rana temporaria , Vejiga Urinaria/efectos de los fármacos , Vasotocina/antagonistas & inhibidores , Vasotocina/farmacología , Agua/metabolismoRESUMEN
During the investigation of the BCG allergenic potency it is advisable to vaccinate with decreasing doses, estimating the lowest dose that induces tuberculin sensitivity and specific morphological inflammation. Although the biological test does not reveal the mathematical correlation of dose-effect relationships, it is important to look for the determination of the minimal sensitizing dose for every BCG vaccine. In this study, three groups of twenty four guinea pigs were vaccinated with decreasing doses of reconstituted BCG vaccine: 120 ng, 12 ng and 1.2 ng. Tuberculin tests were performed in different groups at the 30th, 60th, 90th and 120th day after BCG injection. The negative tuberculin reactions converted to positive between the 60th and 90th day. The dose of 12 ng elicited the largest tuberculin reactions in the animals. This dose contains 65 viable bacteria and could be regarded as the smallest effective sensitizing dose of the BCG vaccine, substrain Sofia SL222. The morphological examination demonstrated that very low inoculums (1.2 ng or 6 viable cells) were sufficient to induce a specific inflammation after BCG vaccination. The immune response in lungs and bronchus-associated lymphoid tissue (BALT) of all BCG doses applied was strongest on the day 60. In the same term, lymph nodes and spleens were characterized with blast transformed lymphoid follicles with epitheloid and Langhans giant cells even with the smallest injected dose of 1.2 ng.
Asunto(s)
Vacuna BCG/administración & dosificación , Hipersensibilidad Tardía/inmunología , Bazo/inmunología , Tuberculosis Pulmonar/inmunología , Vacunación , Animales , Bronquios/inmunología , Bronquios/patología , Relación Dosis-Respuesta Inmunológica , Cobayas , Hipersensibilidad Tardía/patología , Pulmón/inmunología , Pulmón/patología , Activación de Linfocitos/inmunología , Bazo/patología , Tuberculina/inmunología , Tuberculosis Pulmonar/prevención & controlRESUMEN
The effect of the hydraulic retention time (HRT) on the performance of two up-flow anaerobic fixed bed digesters (UFAFBDs) packed with waste tire rubber (D1) and waste tire rubber and zeolite (D2) as micro-organism immobilization supports was studied. It was found that a first-order kinetic model described well the experimental results obtained. The kinetic constants for COD, BOD5, total solids (TS) and volatile solids (VS) removal were determined to be higher in digester D2 than in digester D1 or control. Specifically, they were 0.28 +/- 0.01, 0.32 +/- 0.02, 0.16 +/- 0.01 and 0.24 +/- 0.01 d(- 1) respectively for D1 and 0.33 +/- 0.02, 0.40 +/- 0.02, 0.21 +/- 0.01 and 0.28 +/- 0.01 d(- 1) respectively for D2. This was significant at the 95% confidence level. In addition, the first-order model was also adequate for assessing the effect of the HRT on the removal efficiency and methane production. Maximum methane yield and the first-order constant for methane production were determined and the results obtained were comparable with those obtained by other authors but operating at higher HRTs. Maximum methane yields and the kinetic constant for methane production were 11.1% and 29.4% higher in digester D2 than in D1.
Asunto(s)
Anaerobiosis , Industria Lechera , Estiércol , Zeolitas/química , Reactores Biológicos , CinéticaRESUMEN
We have shown previously that endogenous NO modulates the effect of arginine-vasotocin on the increase in the osmotic water permeability of the frog urinary bladder epithelium. The aim of the present work was to develop a procedure of cultivation of epithelial cells from the frog urinary bladder as a primary culture in order to study in vitro the cellular production of NO and its regulation. Isolated cells were cultivated in modified L-15 medium with 10% FBS and gentamycin (40 microg/ml) at room temperature. Under these conditions, at least 50% cells kept their viability until 8 days of incubation. NO-synthase (NOS) activity was estimated as nitrite (NO2-) accumulation in culture medium; NO2- concentration in the presence of L-NAME, inhibitor of all NOS types, was considered as NOS-independent and was subtracted from each value. The nitrite accumulation was linear in time during 3 days of cultivation and was inhibited by 1400W, inducible NOS (iNOS) inhibitor, and 7-nitroindazole, constitutive NOS's inhibitor, at doses 5-50 and 10-200 microM, respectively. One-day incubation of he cells in the medium with low concentration of gentamycin (1 or 2 microg/ml) led to the significant increase in amount of bacterial in cultured fluid identified as E. coli and Acinetobacter sp. Addition of L-NAME (5 - 103 M) to the medium potentiated the bacteria growth 1.5- and 2.5-times in the presence of 2 and 1 microg gentamycin/ml, respectively. Thus, epithelial cells form the frog urinary bladder possess NO-dependent antibacterial effect which is probably provided by induction of iNOS expression. Taken together, these data demonstrate that the primary culture of the frog urinary bladder epithelial cells is a perspective experimental model for the study of regulation of NOS activity and NO production being of particular interest in relation to the defense effect of NO in epithelia.
Asunto(s)
Técnicas de Cultivo de Célula , Células Epiteliales/citología , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico/biosíntesis , Rana temporaria/metabolismo , Vejiga Urinaria/citología , Acinetobacter/crecimiento & desarrollo , Animales , Supervivencia Celular , Células Cultivadas , Medios de Cultivo/química , Medios de Cultivo/metabolismo , Medios de Cultivo/farmacología , Inhibidores Enzimáticos/farmacología , Células Epiteliales/enzimología , Células Epiteliales/inmunología , Células Epiteliales/microbiología , Escherichia coli/crecimiento & desarrollo , Iminas/farmacología , Indazoles/farmacología , Masculino , Pruebas de Sensibilidad Microbiana , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/análisis , Óxido Nítrico/inmunología , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/inmunología , Vejiga Urinaria/enzimología , Vejiga Urinaria/inmunología , Vejiga Urinaria/microbiologíaRESUMEN
The activity of arginase converting arginine into ornithine and urea is of particular interest among many factors regulating NO production in the cells. It is known that by competing with NO-synthase for common substrate, arginase can affect the NO synthesis. In the present work, the properties of arginase from the frog Rana temporaria L. urinary bladder epithelial cells possessing the NO-synthase activity were characterized, and possible contribution of arginase to regulation of NO production by epithelial cells was studied. It has been shown that the enzyme had the temperature optimum in the range of 55-60 degrees C, K(m) for arginine 23 mM, and V(max) about 10 nmol urea/mg protein/min, and its activity was effictively inhibited by (S)-(2-boronoethyl)-L-cysteine (BEC), an inhibitor of arginase, at concentrations from 10(-6) to 10(-4) M. The comparison of arginase activity in various frog tissues revealed the following pattern: liver > kidney >> brain > urinary bladder (epithelium) > heart > testis. The arginase activity in the isolated urinary bladder epithelial cells was 3 times higher than that in the intact urinary bladder. To evaluate the role of arginase in the regulation of NO production, epithelial cells were cultivated in the media L-15 or 199 containing different amounts of arginine; the concentration of NO2-, the stable NO metabolite, was determined in the culture fluid after 18-20 h of cells incubation. The vast majority of the produced nitrites are associated with the NOS activity, as L-NAME, the NOS-inhibitor, decreased their accumulation by 77.1% in the L-15 medium and by 80% in 199 medium. BEC (10(-4) M) increased the nitrite production by 18.0 % +/- 2.7 in the L-15 medium and by 24.2 +/- 3.5 in the 199 medium (p < 0.05). The obtained data indicate a relatively high arginase activity in the frog urinary bladder epithelium and its involvement in regulation of NO production by epithelial cells.
Asunto(s)
Arginasa/metabolismo , Células Epiteliales/enzimología , Óxido Nítrico/biosíntesis , Vejiga Urinaria/enzimología , Animales , Arginasa/antagonistas & inhibidores , Arginasa/química , Células Cultivadas , Inhibidores Enzimáticos/farmacología , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico/química , Rana temporariaRESUMEN
A study of the role of the depth in the performance of laboratory-scale down-flow anaerobic fixed-bed reactors (DFAFBR) was carried out at different nominal hydraulic retention times (HRT(N)) using piggery waste as substrate at different influent concentrations (2, 4, 6 and 8 g COD/L). The profiles of soluble chemical oxygen demand (COD) (SCOD), organic nitrogen (O.N.), ammonia nitrogen (A.N.), pH and electrical conductivity (E.C.) through the reactor depths showed an initial highly active zone, which was located around the first half of the reactor depth, and a second zone with a lower biological activity. It was found that the depth of the active zone decreased as the HRT(N) increased and that the slopes of the profiles obtained increased with the rise in the influent concentration. A hydraulic test showed an increase in the dispersion number when the HRT(N) increased. The reactors showed a hydraulic pattern between plug-flow and back-mix. The real values of HRT (Theta) also defined as real contact times were determined to be 0.7, 2.1, 3.4, 4.7, 6.4 and 8 days for values of HRT(N) of 1, 2, 3, 4, 5 and 6 days, respectively. It was found that the concentration of SCOD within the reactor decreased exponentially with the increase in the value of theta. Additionally, the influent concentration had a strong influence on the SCOD variation concentration, mainly at values of theta under 1.5 days, which corresponded to the first part of the reactors.
Asunto(s)
Bacterias Anaerobias/fisiología , Reactores Biológicos , Aguas del Alcantarillado/microbiología , Eliminación de Residuos Líquidos/métodos , Anaerobiosis , Animales , Biodegradación Ambiental , Relación Dosis-Respuesta a Droga , Filtración , Concentración de Iones de Hidrógeno , Metano/química , Nitrógeno/química , Oxígeno/química , Aguas del Alcantarillado/química , PorcinosRESUMEN
The influence of the most important variables on the stability and performance of down-flow anaerobic fixed bed reactors treating piggery wastewater after primary sedimentation was evaluated at HRT in the range of 1-6 d and influent substrate concentration in the range of 2 to 12 g TCOD l(-1). The effect of HRT was more pronounced compared to that of influent strength. An increase in the HRT increased the process stability and process performance at different influent strengths. TCOD, SCOD, BOD5 TSS, organic nitrogen (N) and Orthophosphate (P) removals increased with the HRT, independently of the initial substrate concentration (S0). The increase in S0 brought about an increase in the attached biomass concentration (X) at the end of the experiment. Two empirical models based on the individual effect of HRT, X and S0 were evaluated and found to be adequate to describe the influence of these variables on the process performance. The first model took all the above-mentioned variables into consideration while the second model was simplified and based on the use of HRT as the only independent variable. The results obtained by using both models were found to be similar. This demonstrated that independently of the characteristics of the operation, the behaviour and performance of the reactors were comparable. The methane yield coefficient was found to be 0.3371 methane g(-1) TCOD removed.
Asunto(s)
Reactores Biológicos , Estiércol , Aguas del Alcantarillado/microbiología , Eliminación de Residuos Líquidos/métodos , Anaerobiosis , Crianza de Animales Domésticos , Animales , Biodegradación Ambiental , Metano/análisis , Nitrógeno/aislamiento & purificación , Compuestos Orgánicos/aislamiento & purificación , Fosfatos/aislamiento & purificación , Aguas del Alcantarillado/química , PorcinosRESUMEN
In experiments on frog Rana temporaria L. urinary bladder, we investigated localization of NO-synthase (NOS) in urinary bladder slices and measured NOS activity in the suspension of mucosal epithelial cells. Intensive NADPH-diaphorase staining which is widely used as an indicator of NOS activity was found in mucosal epithelium. Almost all mucosal epithelial cells isolated in Ca2+ -free conditions demonstrated positive NADPH-diaphorase reactivity. Direct measurement of NOS activity in suspension of mucosal cells determined by the rate of conversion of L-arginine to L-citrullin showed that the enzyme activity was reduced in absence of external Ca2+ and was inhibited by L-NAME: non-specific NOS inhibitor, and 1400 W: a highly selective iNOS inhibitor (control: 754 +/- 184; L-NAME, 1 mM 329 +/- 87; 1400 W, 20 mM: 547 +/- 25; Ca2+ -free/EDTA: 490 +/- 184 cpm [3H]-citrullin/10(6) cells per 45 min, p < 0.05, n = 7-8). The data obtained demonstrate that frog urinary bladder mucosa epithelial cells provided antidiuretic hormone-induced increase of osmotic water permeability contain nitric oxide synthase. The presence of inducible (iNOS) as well as constitutive isoform(s) revealed in these cells allows to suggest involvement of NOS in intracellular signaling pathways regulated water transport across the epithelium.
Asunto(s)
Células Epiteliales/enzimología , Óxido Nítrico Sintasa de Tipo III/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Óxido Nítrico Sintasa de Tipo I/metabolismo , Vejiga Urinaria/enzimología , Animales , Activación Enzimática , Células Epiteliales/fisiología , Iminas/farmacología , Técnicas In Vitro , Masculino , NADPH Deshidrogenasa/metabolismo , NG-Nitroarginina Metil Éster/farmacología , Óxido Nítrico Sintasa de Tipo I/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo III/antagonistas & inhibidores , Rana temporaria , Vejiga Urinaria/fisiología , Urotelio/enzimología , Urotelio/fisiologíaRESUMEN
The pectic polysaccharide named bergenan BC was obtained using extraction of the green leaves of Siberian tea Bergenia crassifolia (L.) Fritsch. by an aqueous ammonium oxalate. The polysaccharide obtained was proved to comprise mainly d-galacturonic acid, galactose, rhamnose, arabinose and glucose residues and appeared to be pectin. Delayed type hypersensitivity (DTH) reaction to aggregated ovalbumin (agOVA) was found to increase in mice that received bergenan solution (2 mg/mL) for 3 weeks. Bergenan BC was observed to enhance the uptake capacity of human neutrophils at a concentration 100 microg/mL and to stimulate the generation of oxygen radicals by mouse peritoneal macrophages in vitro. Bergenan BC was found to increase the spontaneous adhesion of peritoneal leukocytes and failed to influence adhesion stimulated by PMA or adhesion of peritoneal leukocytes incubated in the presence of 5 mm EDTA. Bergenan failed to show cytotoxic action. The viability of peritoneal leukocytes was estimated to be equal to 91% +/- 8% and 90% +/- 7%% in the control and in the pectin solution at a concentration of 1 mg/mL.Thus, bergenan was shown to possess immunostimulating activity in relation to DTH response in vivo and phagocytic activity in vitro.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Pectinas/farmacología , Saxifragaceae , Animales , Adhesión Celular/efectos de los fármacos , Humanos , Hipersensibilidad Tardía/inmunología , Leucocitos/efectos de los fármacos , Ratones , Neutrófilos/efectos de los fármacos , Pectinas/química , Pectinas/aislamiento & purificación , Cavidad Peritoneal/citología , Fagocitosis/efectos de los fármacos , Extractos Vegetales/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Saxifragaceae/químicaRESUMEN
An oral polybacterial immunomodulator Urostim (U), composed of killed cells and their lysates from E. coli expressing type 1 and Pili, E. coli Rc mutant, P. mirabilis, K. pneumoniae and E. faecalis was created for immunoprophylaxis and immunotherapy of urinary tract infections (UTIs). In experimental animal models, the stimulating effect of U on lymphocyte functional activity, macrophage phagocytosis and antibody producing cells, was established. In this study the immuno-modulating effects of U on the proliferating capacity and ultrastructural morphologic changes of lymphocytes, cytokine production and specific systemic humoral and mucosal immune responses in patients with UTIs have been evaluated. Patients enrolled in the study, received orally 50 mg U daily for a period of three months. On days 0, 30 and 90 a quantitative analysis was performed on lymphoproliferative responses to polyclonal mitogens, IL-2 and the specific antigen U, the production of specific serum and saliva IgA, IgM and IgG antibodies to all components of U and the concentration of pro-inflammatory cytokines. There was significant improvement of non-specific and specific lymphoproliferative responses on days 30 and 90 after the onset of treatment with U, confirmed by electronmicroscopic studies. The highest concentrations of serum proinflammatory cytokines TNF-alpha, IL-1beta and IL-6 were registered at baseline followed by a decrease until the end of the observation period. This finding correlates with the gradual decrease of immune activation as measured by the spontaneous lymphocyte proliferation. Data from the production of specific antibacterial antibodies in serum and saliva show two types of reactions. The first type was registered in patients with low pre-treatment levels in whom the concentration of specific antibodies increased on days 30 and 90. The second type of reaction was observed in patients with high pre-treatment levels, which dropped on day 30 and were usually followed by an increase at the end of the study. These results provide evidence for the immuno-modulating effect of U. Our data show that the oral administration of the polybacterial immunomodulator Urostim stimulates adequate cellular and humoral systemic and mucosal immune responses in patients with chronic UTIs.
Asunto(s)
Adyuvantes Inmunológicos/uso terapéutico , Vacunas Bacterianas/uso terapéutico , Infecciones Urinarias/inmunología , Infecciones Urinarias/terapia , Vacunas Combinadas/uso terapéutico , Adyuvantes Inmunológicos/administración & dosificación , Administración Oral , Adolescente , Adulto , Anciano , Formación de Anticuerpos , Vacunas Bacterianas/administración & dosificación , Técnicas de Cultivo de Célula , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Enfermedad Crónica , Citocinas/biosíntesis , Citocinas/sangre , Femenino , Humanos , Inmunidad Celular , Inmunidad Mucosa , Interleucina-2/farmacología , Linfocitos/efectos de los fármacos , Linfocitos/fisiología , Linfocitos/ultraestructura , Masculino , Persona de Mediana Edad , Mitógenos/farmacología , Factores de Tiempo , Vacunas Combinadas/administración & dosificaciónRESUMEN
On simulating infection caused by different herpes simplex virus type 1 (HSV-1) variants responsive and unresponsive to the drugs acyclovir and phosphonoacetic acid in the cultured Vero and C6 cells has revealed the higher ability of target cells to accumulate 5-aminolevulenic acid (ALA)-induced endogenous porphyrins, which determines the selectivity of their photo damages. Optimal conditions have been defined for all the studied HSV-1 variants to show a virus-inhibiting effect upon photodynamic exposure of infected and ALA-treated cell cultures.
Asunto(s)
Ácido Aminolevulínico/farmacología , Herpes Simple/fisiopatología , Herpesvirus Humano 1/fisiología , Fármacos Fotosensibilizantes/farmacología , Porfirinas/biosíntesis , Replicación Viral/efectos de los fármacos , Replicación Viral/efectos de la radiación , Animales , Chlorocebus aethiops , Herpes Simple/tratamiento farmacológico , Fotoquimioterapia , Ratas , Células VeroAsunto(s)
Neurofibromatosis/diagnóstico , Adulto , Diagnóstico Diferencial , Genes de Neurofibromatosis 1 , Genes de la Neurofibromatosis 2 , Humanos , Imagen por Resonancia Magnética , Masculino , Mutación , Neoplasias de la Vaina del Nervio/diagnóstico , Neoplasias de la Vaina del Nervio/genética , Neurofibromatosis/genética , Neurofibromatosis 1/diagnóstico , Neurofibromatosis 1/genética , Neurofibromatosis 2/diagnóstico , Neurofibromatosis 2/genéticaRESUMEN
Betulonic acid amides with aliphatic and heterocyclic amines and with L-amino acids were synthesized by the acid chloride method. Betulonic acid amide and L-methionine derivatives of betulonic acid and its 3-oxime effectively inhibit the influenza A virus. Betulonic acid octadecylamide is active against the herpes simplex type 1 virus. The conjugate of betulonic acid 3-oxime with L-methionine is also active toward HIV-1. The tested compounds mainly show no activity toward the ECHO6 virus, which is devoid of a coat. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 1; see also http://www.maik.ru.
Asunto(s)
Amidas/química , Aminoácidos/química , Antivirales/síntesis química , Antivirales/farmacología , Ácido Oleanólico/análogos & derivados , Ácido Oleanólico/química , Animales , Células Cultivadas , Embrión de Pollo , VIH-1/efectos de los fármacos , Herpesvirus Humano 1/efectos de los fármacos , Espectroscopía de Resonancia Magnética , Espectrofotometría InfrarrojaRESUMEN
Betulin and betulinic acid have been modified at the C-3 and C-28 positions and the antiviral activity of derivatives has been evaluated in vitro. It was found that simple modifications of the parent structure of lupane triterpenes produced highly effective agents against influenza A and herpes simplex type 1 viruses.
Asunto(s)
Antivirales/farmacología , Triterpenos/farmacología , Antivirales/química , Pruebas de Sensibilidad Microbiana , Triterpenos/químicaRESUMEN
Membranotropic amphiphilic chromophore merocyanine 540 sensitized photodynamic inhibition of drug-resistant and sensitive variants of type I herpes simplex virus in cultured Vero cell. Optimal conditions of photodamage to virus particles and infected cells were determined (merocyanine 540 concentration 1 microM, illumination dose 32.5-65.0 kJ/m(2), exposure at early stages of infection). Infected cells actively bind the photosensitizer, which explains their selective photodamage.
Asunto(s)
Farmacorresistencia Viral , Herpes Simple , Herpesvirus Humano 1/efectos de los fármacos , Fármacos Fotosensibilizantes/farmacología , Pirimidinonas/farmacología , Animales , Antivirales/farmacología , Chlorocebus aethiops , Humanos , Células Vero , Replicación ViralRESUMEN
New nitrogen-containing derivatives of betulinic and betulonic acids, hydrazides and N'-benzalhydrazides, were synthesized. Their antiviral activities toward of influenza A virus, herpes simplex type I virus, enterovirus ECHO6, and HIV-1 were studied in vitro. Betulinic acid 3-oxime was found to have the highest activity against the influenza virus. Betulonic acid, betulinic acid 4-chlorobenzalhydrazide, betulonic acid 3-oxime benzalhydrazide, and betulinic acid hydrazide inhibited the replication of herpes simplex type I virus. Betulinic acid hydrazide also showed antiviral activity toward HIV-1. All the derivatives of betulinic acid under study displayed a low antiviral activity toward enterovirus ECHO6.
Asunto(s)
Antivirales/química , Antivirales/farmacología , Hidrazinas/química , Triterpenos/química , Animales , Antivirales/síntesis química , Bioquímica/métodos , Células Cultivadas/virología , Embrión de Pollo , Evaluación Preclínica de Medicamentos/métodos , Enterovirus/efectos de los fármacos , VIH-1/efectos de los fármacos , Herpesvirus Humano 1/efectos de los fármacos , Humanos , Hidrazinas/síntesis química , Hidrazinas/farmacología , Virus de la Influenza A/efectos de los fármacos , Oximas/síntesis química , Oximas/química , Oximas/farmacología , Triterpenos Pentacíclicos , Relación Estructura-Actividad , Triterpenos/farmacología , Ácido BetulínicoRESUMEN
Antiviral properties of betulin, betulinic and betulonic acids were investigated in cell cultures infected with herpes simplex type I, influenza FPV/Rostock and ECHO 6 viruses. All studied triterpenes were active against herpes simplex virus. Betulin and especially betulinic acid also suppressed ECHO 6 virus reproduction.