RESUMEN
The timely diagnosis and treatment of post-infectious glomerulonephritis (PIGN) is currently limited by the erased and low-symptom nature of the disease, which leads to the search for informative biological markers of the disease, which can be used as immunological indicators of blood and urine. The study was carried out in order to establish the characteristic changes in the immunological parameters of blood and urine in patients with PIGN. The study included 60 patients with PIGN from among the patients, hospitalized in the nephrology department of the Republican Clinical Hospital of Health Care Ministry of the Chuvash Republic in 2015-2018. In addition to the generally accepted research methods, the patients underwent: 1) the determination of indicators of innate and acquired immune response in the blood (CD3+ -, CD3+ CD4+-, CD3+CD8+-, CD4+CD25+-, CD95+-, CD20+-, CD14+CD282+-, CD14+CD284+- cells; levels of IgG, IgA, IgM, circulating immune complexes, C3, C4) and urine (levels of IgG, IgA, IgM, C3, C4); 2) the determination of the levels of cytokines - IL-1ß, Ra-IL-1ß, IL-2, IL-4, IL-8, IL-10, IL-17A in blood serum and urine. The data obtained were compared with those of the group of healthy individuals. The changes in blood immunological parameters, identified in the group of patients with PIGN, indicate the activation of innate immunity (the increase in the number of CD14+TLR2+- cells) and the humoral component of adaptive immunity (the increase in the number of B-lymphocytes, hyperimmunoglobulinemia - the increase in IgM and IgA levels) against the background of the decrease in the number of T (CD3+) - lymphocytes and regulatory (CD4+CD25high) - cells, hypocomplementemia (decreased levels of C3, C4). The increase in the content of C3, IgG and IgA was found in the urine. The cytokine profile of blood in patients with PIGN was characterized by the increase in the levels of pro- and anti-inflammatory cytokines IL-1ß, Ra-IL-1ß, IL-2, IL-8, IL-10, IL-17A, with the exception of IL-4, which remained on the levels of healthy individuals. The cytokine profile of urine in patients was characterized by the increase in the levels of pro-inflammatory cytokines IL-1ß, IL-2, IL-8, IL-17A and anti-inflammatory cytokine - IL-10, with no changes in the content of Ra-IL-1ß and IL-4. The revealed features of the immunological profile of blood and urine in patients with PIGN reflect the immunopathogenetic mechanisms of this disease.
Asunto(s)
Líquidos Corporales , Glomerulonefritis , Linfocitos B , Citocinas , HumanosRESUMEN
Over the past decade, there has been a great interest in hematological indicators as markers for assessing the severity of coronary artery disease and stratifying the risk of cardiovascular events in patients with acute coronary syndrome. To understand the possibility of using hematological indicators as predictors of adverse outcomes in this category of patients, the question of clarifying their age-related changes became a question. This study discusses age-related changes in hematological parameters in patients with acute coronary syndrome.
Asunto(s)
Síndrome Coronario Agudo , Enfermedad de la Arteria Coronaria , Síndrome Coronario Agudo/diagnóstico , Síndrome Coronario Agudo/epidemiología , Biomarcadores , Humanos , Factores de RiesgoRESUMEN
We studied cytotoxic activity of new tetranitrosyl NO-generating binuclear iron-sulfur [Fe-S] complexes containing different ligands in the molecule against tumor cells in vitro. Cytotoxic activity of the most active complex with cysteamine (CysAm) was compared with that of antitumor drug cisplatin. Caspase activation and morphological changes in cells were visualized by fluorescence microscopy. Fluorescence of active caspases 3 and 7 and changes in nuclear DNA in cells in the presence of CyAm were detected by using fluorochrome-labeled inhibitor of caspases (FLICA) and Hoechst and propidium iodide reagents. Similar cytotoxic activities of CyAm and cisplatin were demonstrated in various human tumor cell lines of different histogenesis. Therefore, a new class of NO-donating [Fe-S] complexes can provide the base of potential drugs for chemotherapy with a new mechanism of action.
Asunto(s)
Antineoplásicos/farmacología , Hierro/farmacología , Óxidos de Nitrógeno/farmacología , Azufre/farmacología , Células A549 , Antineoplásicos/química , Línea Celular Tumoral , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Hierro/química , Células K562 , Células MCF-7 , Óxidos de Nitrógeno/química , Azufre/químicaRESUMEN
The episode of acute coronary syndrome is most often preceded by the development of systemic and local inflammation, which plays a significant role in the pathogenesis of the disease. General clinical blood analysis, directly or indirectly reflecting systemic pathological processes in the patient's body based on quantitative and morphological assessment of blood composition, is one of the most affordable methods of laboratory diagnostics in modern public health. Taking into account the growing number of digital data obtained by diagnosticians from analytical systems, there is a growing potential for the use of machine learning methods to increase the effectiveness of provided diagnostic information in the interests of the patient. The aim of this study was to create an algorithm for stratifying the risk of myocardial infarction based on the methods of machine learning in patients with acute coronary syndrome at primary examination. A prospective pilot study was conducted. In total 307 patients with acute coronary syndrome (169 men and 138 women) were examined. The average age of patients was 68.6 ± 12.5 years. Retrospectively, the patients were divided into two groups: the main group - patients with the final diagnosis "Myocardial infarction" and the control group with the diagnosis "Unstable angina pectoris". All patients at hospitalization at the primary laboratory examination along with the study of the concentration of cardiac troponin I by a highly sensitive method were examined by a general clinical bloodanalysis on an automatic hematological 5-diff analyzer. As a result of the application of the ensemble method as a method of machine learning and artificial neural networks as 6 independent models of the ensemble it was possible to achieve the area under the ROC curve = 0.77 on the test set when assessing the quality of patient stratification. Taking into account the volume of the training sample in 214 patients and the results of similar studies, the achieved stratification quality can be considered acceptable and promising for further accumulation of the database with the purpose of additional training of the developed algorithm and improvement of the disease prognosis accuracy characteristics.
Asunto(s)
Síndrome Coronario Agudo/complicaciones , Infarto del Miocardio/diagnóstico , Anciano , Anciano de 80 o más Años , Algoritmos , Biomarcadores , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proyectos Piloto , Estudios Prospectivos , Factores de RiesgoRESUMEN
We studied specific antitumor activity of a liposomal drug based on tetra-3-phenylthiophthalocyanine aluminum hydroxide (lipophthalocyan) intended for photodynamic therapy. The optimal dose and protocol for photodynamic therapy with lipophthalocyan were chosen in experiments on mice: single intravenous dose of 6 mg/kg with a 5-h interval between administration and laser exposure and irradiation energy density of 400 J/cm2. A wide spectrum antitumor activity of lipophthalocyan was demonstrated in vivo for various transplantable mouse tumors (Lewis lung epidermoid carcinoma, S37 sarcoma, and colon adenocarcinoma AKATOL). The results show the possibility of using lipophthalocyan for photodynamic therapy of tumors of surface localization (skin and mucosa tumors).
Asunto(s)
Rayos Láser , Liposomas/química , Fotoquimioterapia/métodos , Fármacos Fotosensibilizantes/uso terapéutico , Animales , Carcinoma Pulmonar de Lewis/terapia , Neoplasias del Colon/terapia , Femenino , Masculino , Ratones , Ratones Endogámicos BALB C , Sarcoma/terapiaRESUMEN
The syndrome of senile asthenia (frailty) in patients with acute coronary syndrome is recognized as one of the risk factors for the development of adverse outcomes after undergoing cardiosurgical interventions, and the search for an optimal method for assessing this syndrome could be a screening to identify patients at increased risk. This study compares two methods for assessing senile asthenia syndrome in patients with acute coronary syndrome, its prevalence in this category of patients, its relationship with clinical characteristics and 1-year mortality after hospitalization. According to the results of the study, senile asthenia syndrome was observed more often in the group of patients older than 75 years. Patients with senile asthenia assessment using the Green scale had a higher risk on the GRACE scale and the presence of this syndrome was an independent 1-year mortality predictor after hospitalization.
Asunto(s)
Síndrome Coronario Agudo , Fragilidad , Síndrome Coronario Agudo/complicaciones , Síndrome Coronario Agudo/diagnóstico , Síndrome Coronario Agudo/epidemiología , Humanos , Prevalencia , Pronóstico , Medición de Riesgo , Factores de RiesgoRESUMEN
We performed an in vivo comparative study of activity of three substances of the nitrosourea group produced in Russia. All substances demonstrated high antitumor activity against various solid and leukemic tumors. Aranosa significantly enhanced life duration in mice with leukemia (by 65-194%) and inhibited the growth of solid tumors (by 49-99.6%). Lisomustine and ormustine showed higher activity than aranose. Single administration of lisomustine increased life span of mice (by 22-114%) and resulted in cure of all animals in four models: lymphoblastic leukemia L-1210, lymphocytic leukemia P-388, Lewis lung carcinoma, and cervical cancer RShM-5. After ormustine treatment, full recovery was observed only in groups with lymphocytic leukemia P-388 and cervical cancer RShM-5. These findings attest to higher activity of lisomustine in the studied models.
Asunto(s)
Antineoplásicos/uso terapéutico , Compuestos de Nitrosourea/uso terapéutico , Animales , Femenino , Leucemia L1210/tratamiento farmacológico , Leucemia P388/tratamiento farmacológico , Masculino , Ratones , Neoplasias Experimentales/tratamiento farmacológico , Federación de Rusia , Resultado del Tratamiento , Neoplasias del Cuello Uterino/tratamiento farmacológicoRESUMEN
The purpose of study is to examine associations of polymorphic allele genes HLA class II - DRB1, DQA1 and DQB1 and their haplotypic combinations with chronic obstructive disease of lungs in workers of organocilicic manufacture in Chuvash population. The HLA-genotyping was implemented to 50 patients with chronic obstructive disease of lungs and 38 healthy workers of organocilicic manufacture, belonging to Chuvash ethnic population. The genotyping was implemented on three genes HLA: DRB1 (14 alleles), DQA1 (8 alleles) and DQB1 (11 alleles) using multi-primary polymerase chain reaction technique. The typing was implemented using kits manufactured by "DNA-Technology" (Russia). The degree of association ofHLA-alleles and gaplotypes with development of chronic obstructive disease of lungs in conditions of organocilicic manufacture was determined by value of relative risk (rr). The study established negative associations of chronic obstructive disease of lungs with alleles HLA-DRB1*01 (rr=0.021; p<0.001); DQA1*0101 (rr=0.013; p<0.001); DQB1*0501 (rr=0.021; p<0.001) andgaplotypes DRB1*01-DQA1*0101 (rr=0.031; p<0.01); DRB1*07-DQA1*0201 (rr=0.076; p<0.01); Bm1*13-DQA1*0102 (rr=0.11; p<0.05). The HLA-markers of resistance to development of chronic obstructive disease of lungs in conditions of organocilicic manufacture in Chuvash population.
Asunto(s)
Cadenas alfa de HLA-DQ/genética , Cadenas beta de HLA-DQ/genética , Cadenas HLA-DRB1/genética , Enfermedad Pulmonar Obstructiva Crónica/genética , Adulto , Anciano , Alelos , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Técnicas de Genotipaje , Haplotipos/genética , Humanos , Masculino , Persona de Mediana Edad , Exposición Profesional , Enfermedad Pulmonar Obstructiva Crónica/inducido químicamente , Enfermedad Pulmonar Obstructiva Crónica/epidemiología , Enfermedad Pulmonar Obstructiva Crónica/patología , Federación de Rusia , Silicio/toxicidadRESUMEN
OBJECTIVE: To evaluate the efficacy of surgical treatment of lumbar vertebral canal stenosis based on the study of patient's pain and emotional status, quality of life and level of disability before and after surgical treatment. MATERIAL AND METHODS: A complex examination of 163 patients with lumbar vertebral canal stenosis was carried out. The visual analogous scale, Paindetect (PD) and Douleur neuropathique 4 (DN4), the Chronic Pain Coping Inventory, the Oswestry low back pain disability questionnaire were used. Affective disorders were measured using Beck depression scale. Blood serum and thrombocytic serotonin were determined with ELISA. Patients were examined at admission before the surgery, at discharge from the hospital after surgical treatment and one year after the surgery. RESULTS AND CONCLUSION: Before surgery, the patients had chronic pain syndrome of mixed etiology (nociceptive and neurotic type) and neurological disorders that significantly affected quality of life and work capacity. Neuropathic pain and depression, reduced quality of life and daily activities were more pronounced in patients with central canal stenosis compared to patients with lateral stenosis. The timely surgical treatment of lumbar vertebral canal stenosis promoted the decompression of nervous structures. A significant reduction of pain syndrome and the level of depression as well as the improvement of quality of life were observed.
Asunto(s)
Laminectomía , Dolor de la Región Lumbar/psicología , Vértebras Lumbares/cirugía , Dolor Postoperatorio/psicología , Calidad de Vida , Fusión Vertebral , Estenosis Espinal/cirugía , Adolescente , Adulto , Anciano , Femenino , Estudios de Seguimiento , Humanos , Dolor de la Región Lumbar/diagnóstico , Dolor de la Región Lumbar/etiología , Masculino , Persona de Mediana Edad , Dimensión del Dolor , Dolor Postoperatorio/diagnóstico , Periodo Preoperatorio , Estenosis Espinal/complicaciones , Adulto JovenRESUMEN
UNLABELLED: The purpose of the study was the assessment of the effectiveness of vaccination against diphtheria in the Voronezh region over the epidemic period of 1993-1997 and epidemiological welfare during 2010-2014. MATERIALS AND METHODS: of the study: data of the official statistical reporting--forms number 1, 6, the serum level of antitoxic antibodies to diphtheria in 19319 healthy individuals were analyzed with the aid of epidemiological (descriptive and evaluative), immunological and statistical methods. RESULTS: During the epidemic rise of diphtheria (1993-1997) 75% of cases were amounted to the adult population of the Voronezh region, half of them--were not immunized against diphtheria. In 1993 there was begun mass vaccination of adult population, immunization coverage by 1998 reached 95%. According to seromonitoring data the share of seronegatives to diphtheria among cases examined during the period of 1995-2000 accountedfrom 11.9 to 24.9%. During the period of sporadic morbidity (1998-2007 years) among patients the 80% of cases have been vaccinated with an interval from the last inoculation of 3-5 years, which casts doubt on the effectiveness of vaccines. Since 2008 the incidence of diphtheria in the Voronezh region was not recorded. Against the background of 98% coverage of vaccination of the total population, the share of seronegatives for the last 5 years have decreased by 2.5 times and in 2014 reached the required performance. CONCLUSION: Documented inoculation indices fail to reflect the level of the actual protection against infection. In the conditions of the absence of the morbidity only serological monitoring is an objective criterion of the protectability of the population from infection.
Asunto(s)
Difteria/prevención & control , Vacunación Masiva/organización & administración , Vigilancia de la Población , Población Rural , Adulto , Difteria/epidemiología , Femenino , Humanos , Incidencia , Masculino , Federación de RusiaRESUMEN
Cytestrole acetate (CA), in the structure of which the steroidal antiestrogen component is associated with bis-ß-cloroethylamino group, exhibits a strong cytotoxic activity against hormone-dependent cancer cell lines (CaOV, HeLa, MCF-7). In doxorubicin-resistant MCF-7 cells, CA potentiates the cytotoxic effect of etoposide and doxorubicin, and the IC50 for CA in these cells is 40 times lower than that for tamoxifen (TAM). In transplantable mice breast adenocarcinoma Ca-755, the therapeutic CA dose is 25 mg/kg when administered subcutaneously in oil solution for 5 days. On the DMBA-induced mammary tumors in rats, CA injected subcutaneously led to partial regressions 4 weeks after treatment in 75% of test rats, whereas TAM produced this effect in 43% of rats. Among various drug forms of CA, the most active were oil solution of CA in gelatin capsules for oral use and liposomal emulsion for intravenous administration, since these forms exhibited the highest values of Ca-755 tumor growth inhibition index (TGI = 97 - 98%).
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Antineoplásicos Hormonales/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Citostáticos/farmacología , Antagonistas de Estrógenos/farmacología , Etinilestradiol/análogos & derivados , Adenocarcinoma/patología , Animales , Neoplasias de la Mama/patología , Línea Celular Tumoral , Doxorrubicina/farmacología , Esquema de Medicación , Sinergismo Farmacológico , Etinilestradiol/farmacología , Etopósido/farmacología , Femenino , Humanos , Concentración 50 Inhibidora , Inyecciones Subcutáneas , Ratones , Ratas , Tamoxifeno/farmacología , Carga Tumoral/efectos de los fármacos , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
The efficiency of photodynamic therapy with photosensitizer Tiosens (Russia) was evaluated in mono- and combined therapy of rats with malignant gliomas (glioblastoma 101/8, oligodendroglioma 14-4-9, and C6 glioma). The efficiency of photodynamic monotherapy was not high: the animals died from brain edema developing in tumor tissue and in the adjacent normal cerebral tissue. Pathomorphological studies of tumor tissue detected necrosis and apoptosis, destruction of vessels with hemorrhages, and vascular thrombosis. Combined therapy for malignant gliomas including Tiosens photodynamic therapy and subsequent temodal or lysomustine chemotherapy, was the most effective. In glioblastoma 101/8, combined therapy with lysomustine or temodal led to prolongation of the lifespan by 127 %; 62.5 and 50 % rats were cured, respectively; in oligodendroglioma 14-4-9, animal lifespan was prolonged by 80 and 60 %, with 43 and 45 % rats cured, respectively. Glioma C6 was least sensitive to therapy.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Neoplasias Encefálicas/tratamiento farmacológico , Glioblastoma/tratamiento farmacológico , Oligodendroglioma/tratamiento farmacológico , Fotoquimioterapia , Animales , Animales no Consanguíneos , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Dacarbazina/administración & dosificación , Dacarbazina/análogos & derivados , Ensayos de Selección de Medicamentos Antitumorales , Humanos , Indoles/administración & dosificación , Trasplante de Neoplasias , Compuestos de Nitrosourea/administración & dosificación , Compuestos Organometálicos/administración & dosificación , Fármacos Fotosensibilizantes/administración & dosificación , Ratas Wistar , TemozolomidaRESUMEN
The results of microbiological tests of 2382 blood specimens from 698 patients of three ICUs of the N. V. Sklifosovsky Research Institute of Emergency Service were analysed for a period of 2009 to 2012. No microbial growth was detected in 1160 specimens (48.7%). In the subsequent tests 816 isolates were used. The quota of grampositive isolates in the pathogen pattern decreased from 51.68 to 35.9% and that of gramnegative isolates increased from 38.26 to 48.1%. The number of the Candida isolates increased from 7.38 to 15.2%. The frequency of the S. aureus and Enterococcus spp. isolates lowered from 26.2 to 13.3% and from 15.4 to 11.5% respectively. The quota of K. pneumoniae isolates and Acinetobacter spp. increased from 8.1 to 18.9% and from 14.1 to 17.4% respectively. Most of the S. aureus isolates were methicillin resistant. All the staphylococcal isolates were susceptible to vancomycin and linezolid. All the strains of K. pneumoniae isolated in 2009-2012 produced extended spectrum beta-lactamases. In 2012 7.8% the K. pneumoniae isolates were resistant to carbapenems. In 2012 25.5% of the Acinetobacter spp. isolates were susceptible to carbapenems and 70.2% were susceptible to cefoperazon/sulbactam.
Asunto(s)
Bacteriemia/microbiología , Candidiasis/microbiología , Infecciones por Bacterias Gramnegativas/microbiología , Infecciones por Bacterias Grampositivas/microbiología , Sepsis/microbiología , Acetamidas/uso terapéutico , Acinetobacter/efectos de los fármacos , Acinetobacter/crecimiento & desarrollo , Antibacterianos/uso terapéutico , Bacteriemia/tratamiento farmacológico , Candida/efectos de los fármacos , Candida/crecimiento & desarrollo , Candidiasis/tratamiento farmacológico , Carbapenémicos/uso terapéutico , Cefoperazona/uso terapéutico , Farmacorresistencia Bacteriana Múltiple , Enterococcus/efectos de los fármacos , Enterococcus/crecimiento & desarrollo , Infecciones por Bacterias Gramnegativas/tratamiento farmacológico , Infecciones por Bacterias Grampositivas/tratamiento farmacológico , Humanos , Unidades de Cuidados Intensivos , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/crecimiento & desarrollo , Linezolid , Meticilina/uso terapéutico , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Staphylococcus aureus Resistente a Meticilina/crecimiento & desarrollo , Oxazolidinonas/uso terapéutico , Sepsis/tratamiento farmacológico , Sulbactam/uso terapéutico , Vancomicina/uso terapéutico , beta-LactamasasRESUMEN
The aim of the work was to study the state of adaptive systems in patients with metabolic syndrome. This cross-sectional randomized trial included 99 women and 32 men. The measured variables were blood levels of dehydroepiandrosterone sulfate (DGEAS), cortisol, total T3 and T4, lymphocyte count, and muscle endurance (number of squats/min). 43 patients presented with metabolic syndrome (MS) and 88 without it. MS was associated with decreased blood DGEAS and T3 levels, increased lymphocyte count, reduced activity of sympathetic nervous system and muscle endurance.
Asunto(s)
Adaptación Fisiológica , Sulfato de Deshidroepiandrosterona/sangre , Tolerancia al Ejercicio , Síndrome Metabólico , Sistema Nervioso Simpático/fisiopatología , Adulto , Índice de Masa Corporal , Estudios Transversales , Modificador del Efecto Epidemiológico , Femenino , Humanos , Hidrocortisona/sangre , Recuento de Linfocitos , Masculino , Síndrome Metabólico/epidemiología , Síndrome Metabólico/metabolismo , Síndrome Metabólico/fisiopatología , Persona de Mediana Edad , Distribución Aleatoria , Factores de Riesgo , Triyodotironina/sangreRESUMEN
The microbiological tests of 769 blood samples from 220 patients, treated in 4 intensive care units of the N.V. Sklifosovsky Research Institute for Emergency Medical Service within a period from January 2009 to June 2010, were analysed. Etiologically significant microorganisms were detected in 323 samples (42%). 253 isolates were used in the analysis. Grampositive and gramnegative pathogens were detected in 47 and 42% of the cases respectively. Candida and anaerobic organisms were isolated in 8 and 3% of the cases respectively. Staphylococcus aureus and enterococci were isolated in 24 and 15% of the cases respectively. Nonfermenting gramnegative bacteria and enterobacteria were revealed in 25 and 17% of the cases respectively. Differences in the spectrum of the sepsis pathogens depending on the patients contingent were shown. The maximum summary susceptibility of the grampositive cocci was observed with respect to vancomycin and linezolid and that of the gramnegative bacteria was stated with respect to imipenem and meropenem.
Asunto(s)
Bacterias Anaerobias , Candida , Bacterias Gramnegativas , Bacterias Grampositivas , Unidades de Cuidados Intensivos/estadística & datos numéricos , Acetamidas/farmacología , Acetamidas/uso terapéutico , Antibacterianos/farmacología , Antibacterianos/uso terapéutico , Bacterias Anaerobias/efectos de los fármacos , Bacterias Anaerobias/aislamiento & purificación , Infecciones Bacterianas/tratamiento farmacológico , Infecciones Bacterianas/etiología , Candida/efectos de los fármacos , Candida/aislamiento & purificación , Infección Hospitalaria/tratamiento farmacológico , Infección Hospitalaria/etiología , Infección Hospitalaria/microbiología , Farmacorresistencia Microbiana , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Gramnegativas/aislamiento & purificación , Bacterias Grampositivas/efectos de los fármacos , Bacterias Grampositivas/aislamiento & purificación , Humanos , Imipenem/farmacología , Imipenem/uso terapéutico , Control de Infecciones/normas , Linezolid , Meropenem , Moscú , Oxazolidinonas/farmacología , Oxazolidinonas/uso terapéutico , Sepsis/tratamiento farmacológico , Sepsis/etiología , Sepsis/microbiología , Tienamicinas/farmacología , Tienamicinas/uso terapéutico , Vancomicina/farmacología , Vancomicina/uso terapéuticoRESUMEN
In endothelial cells there remain uncertainties in the details of how Ca(2+) signals are generated and maintained, especially in intact preparations. In particular the role of the sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA), in contributing to the components of agonist-induced signals is unclear. The aim of this work was to increase understanding of the detailed mechanism of Ca(2+) signalling in endothelial cells using real time confocal imaging of Fluo-4 loaded intact rat tail arteries in response to muscarinic stimulation. In particular we have focused on the role of SERCA, and its interplay with capacitative Ca(2+) entry (CCE) and ER Ca(2+) release and uptake. We have determined its contribution to the Ca(2+) signal and how it varies with different physiological stimuli, including single and repeated carbachol applications and brief and prolonged exposures. In agreement with previous work, carbachol stimulated a rise in intracellular Ca(2+) in the endothelial cells, consisting of a rapid initial phase, then a plateau upon which oscillations of Ca(2+) were superimposed, followed by a decline to basal Ca(2+) levels upon carbachol removal. Our data support the following conclusions: (i) the size (amplitude and duration) of the Ca(2+) spike and early oscillations are limited by SERCA activity, thus both are increased if SERCA is inhibited. (ii) SERCA activity is such that brief applications of carbachol do not trigger CCE, presumably because the fall in luminal Ca(2+) is not sufficient to trigger it. However, longer applications sufficient to deplete the ER or even partial SERCA inhibition stimulate CCE. (iii) Ca(2+) entry occurs via STIM-mediated CCE and SERCA contributes to the cessation of CCE. In conclusion our data show how SERCA function is crucial to shaping endothelial cell Ca signals and its dynamic interplay with both CCE and ER Ca releases.
Asunto(s)
Arterias/citología , Señalización del Calcio , Calcio/agonistas , Células Endoteliales/metabolismo , Animales , Compuestos de Boro/farmacología , Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Carbacol/farmacología , Forma de la Célula/efectos de los fármacos , Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/metabolismo , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Células Endoteliales/enzimología , Estrenos/farmacología , Espacio Extracelular/efectos de los fármacos , Espacio Extracelular/metabolismo , Indoles/farmacología , Lantano/farmacología , Glicoproteínas de Membrana/metabolismo , Microscopía Confocal , Pirrolidinonas/farmacología , Ratas , Rianodina/farmacología , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/antagonistas & inhibidores , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismo , Molécula de Interacción Estromal 1 , Factores de Tiempo , Fosfolipasas de Tipo C/metabolismoRESUMEN
Recent studies have highlighted the role of the sarcoplasmic reticulum (SR) in controlling excitability, Ca2+ signalling and contractility in smooth muscle. Caffeine, an agonist of ryanodine receptors (RyRs) on the SR has been previously shown to effect Ca2+ signalling but its effects on excitability and contractility are not so clear. We have studied the effects of low concentration of caffeine (1 mM) on Ca2+ signalling, action potential and contractility of guinea pig ureteric smooth muscle. Caffeine produced reversible inhibition of the action potentials, Ca2+ transients and phasic contractions evoked by electrical stimulation. It had no effect on the inward Ca2+ current or Ca2+ transient but increased the amplitude and the frequency of spontaneous transient outward currents (STOCs) in voltage clamped ureteric myocytes, suggesting Ca2+-activated K+ channels (BK) are affected by it. In isolated cells and cells in situ caffeine produced an increase in the frequency and the amplitude of Ca2+ sparks as well the number of spark discharging sites per cell. Inhibition of Ca2+ sparks by ryanodine (50 microM) or SR Ca2+-ATPase (SERCA) cyclopiazonic acid (CPA, 20 microM) or BKCa channels by iberiotoxin (200 nM) or TEA (1 mM), fully reversed the inhibitory effect of caffeine on Ca2+ transients and force evoked by electrical field stimulation (EFS). These data suggest that the inhibitory effect of caffeine on the action potential, Ca2+ transients and force in ureteric smooth muscle is caused by activation of Ca2+ sparks/STOCs coupling mechanism.
Asunto(s)
Cafeína/farmacología , Señalización del Calcio/fisiología , Músculo Liso , Uréter/anatomía & histología , Potenciales de Acción/efectos de los fármacos , Potenciales de Acción/fisiología , Animales , Canales de Calcio Tipo L/metabolismo , Femenino , Cobayas , Indoles/metabolismo , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Músculo Liso/efectos de los fármacos , Músculo Liso/fisiología , Técnicas de Placa-Clamp , Inhibidores de Fosfodiesterasa/farmacología , Canales de Potasio Calcio-Activados/metabolismo , Rianodina/metabolismo , Retículo Sarcoplasmático/metabolismoRESUMEN
The data on paraendocervical administration of metronidazole in the treatment of patients with relapsing Trichomonas endocervicitis are presented. Metronidazole was administered as 0.5% solution in a dose of 0.04 g once a day for 8-10 days in complex traditional therapy including oral use of metronidazole and immunocorrigating and local treatment. It was shown that paraendocervical administration of the protistocidal agent provided earlier regression of the urogenital symptoms and 2.8 times lower frequency of the relapses.
Asunto(s)
Antitricomonas/uso terapéutico , Metronidazol/uso terapéutico , Vaginitis por Trichomonas/diagnóstico , Cervicitis Uterina/tratamiento farmacológico , Administración Intravaginal , Adulto , Animales , Antitricomonas/administración & dosificación , Femenino , Humanos , Metronidazol/administración & dosificación , Cervicitis Uterina/parasitologíaRESUMEN
Recent data have shown Ca(2+)-dependent activation of Rho-kinase by sustained depolarization of arterial smooth muscle. Visceral smooth muscles, however, contract phasically in response to action potentials and it is unclear whether Ca(2+)-dependent or -independent Rho-kinase activation occurs. We have therefore investigated this, under physiologically relevant conditions, in intact ureter. Action potentials, ionic currents, Ca(2+) transients, myosin light chain (MLC) phosphorylation and phasic contraction evoked by action potentials in guinea-pig and rat ureter were investigated. In rat, but not guinea-pig ureter, three Rho-kinase inhibitors, Y-27632, HA-1077 and H-1152, significantly decreased phasic contractions and Ca(2+) transients. Voltage- and current-clamp data showed that Rho-kinase inhibition reduced the plateau component of the action potential, inhibited Ca(2+)-channels and, indirectly, Ca(2+)-activated Cl(-) channels. The Ca(2+) channel agonist Bay K8644 could reverse these effects. The K(+) channel blocker TEA could also reverse the inhibitory effect of Y-27632 on the action potential and Ca(2+) transient. Ca(2+) transients and inward current, activated by carbachol-induced sarcoplasmic reticulum Ca(2+)release, were not affected by Rho-kinase inhibition. Rho-kinase inhibition produced a Ca(2+)-independent increase in the relaxation rate of contraction, associated with acceleration of MLC dephosphorylation, which was sensitive to calyculin A. These data show for the first time that: (1) Rho-kinase has major effects on Ca(2+) signalling associated with the action potential, (2) this effect is species dependent and (3) Rho-kinase controls relaxation of phasic contraction of myogenic origin. Thus Rho-kinase can modulate phasic smooth muscle in the absence of agonist, and the mechanisms are both Ca(2+)-dependent, involving ion channels, and Ca(2+)-independent, involving MLC phosphorylation activity.
Asunto(s)
Potenciales de Acción/fisiología , Señalización del Calcio/fisiología , Músculo Liso/fisiología , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , Uréter/fisiología , Potenciales de Acción/efectos de los fármacos , Animales , Canales de Calcio/fisiología , Señalización del Calcio/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Cobayas , Técnicas In Vitro , Péptidos y Proteínas de Señalización Intracelular , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Relajación Muscular/efectos de los fármacos , Relajación Muscular/fisiología , Músculo Liso/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/metabolismo , Ratas , Uréter/efectos de los fármacos , Quinasas Asociadas a rhoRESUMEN
In experiments carried out with the use of the radioactive label (45Ca2+) on suspension of the rat uterus myocytes processed by digitonin solution (0.1 mg/ml), influence of spermine and cyclosporin A on Mg2+, ATP-dependent Ca2+ transport in mitochondria at different Mg2+ concentration were investigated. Ca2+ accumulation in mitochondria was tested as such which was not sensitive to thapsigargin (100 nM) and was blocked by ruthenium red (10 microM). It has been shown, that spermine (1 mM) stimulates Mg2+, ATP-dependent Ca2+ accumulation in mitochondria irrespective of Mg2+ concentration (3 or 7 mM) in the incubation medium. At the same time cyclosporin A (5 microM) effects on Ca2+ accumulation in mitochondria depend on Mg2+ concentration in the incubation medium: at 3 mM Mg2+ the stimulating effect was observed, and at 7 mM Mg2+ - the inhibitory one. In conditions which led to the increase of nonspecific mitochondrial permeability and, accordingly, to dissipation of electrochemical potential (it was reached by 5 min. preincubation of myocytes suspension in the medium that contained 10 microM Ca2+, 2 mM phosphate and 3 or 7 mM Mg2+, but not ATP) significant inhibition of Mg2+, ATP-dependent Ca2+ accumulation in mitochondria was observed. The inhibition to the greater degree was observed when medium ATP and Mg2+ were absent simultaneously in the preincubation. Thus the quality of spermine effects on Ca2+ accumulation was kept: stimulation in the presence both of 3 mM and 7 mM Mg2+. Ca2+ accumulation did not reach the control level when 3 mM Mg2+ and 1 mM spermine was present and ATP absent in the preincubation medium. However, in the presence of 7 mM Mg2+ and 1 mM spermine practically full restoration (up to a control level) of Ca2+ accumulation was observed. At the same time with other things being equal such restoration was not observed at simultaneous absence of ATP and Mg2+ in the preincubation medium. The quality of cyclosporin A effects on Ca2+ accumulation in mitochondria was also kept: stimulation - in the presence of 3 mM Mg2+, inhibition - in the presence of 7 mM Mg2+ in the preincubation medium. And, at last, in the presence of cyclosporin A irrespective of the fact which preincubation medium was used, Ca2+ accumulation level practically did not depend on Mg2+ concentration.