ABSTRACT
The cardiac perivascular niche is a cellular microenvironment of a blood vessel. The principles of niche regulation are still poorly understood. We studied the effect of TGFß1 on cells forming the cardiac perivascular niche using 3D cell culture (cardiospheres). Cardiospheres contained progenitor (c-Kit), endothelial (CD31), and mural (αSMA) cells, basement membrane proteins (laminin) and extracellular matrix proteins (collagen I, fibronectin). TGFß1 treatment decreased the length of CD31+ microvasculature, VE cadherin protein level, and proportion of NG2+ cells, and increased proportion of αSMA+ cells and transgelin/SM22α protein level. We supposed that this effect is related to the stabilizing function of TGFß1 on vascular cells: decreased endothelial cell proliferation, as shown for HUVEC, and activation of mural cell differentiation.
Subject(s)
Cell Differentiation , Cell Proliferation , Transforming Growth Factor beta1 , Transforming Growth Factor beta1/pharmacology , Transforming Growth Factor beta1/metabolism , Cell Differentiation/drug effects , Humans , Cell Proliferation/drug effects , Human Umbilical Vein Endothelial Cells/metabolism , Human Umbilical Vein Endothelial Cells/drug effects , Animals , Microfilament Proteins/metabolism , Microfilament Proteins/genetics , Platelet Endothelial Cell Adhesion Molecule-1/metabolism , Cadherins/metabolism , Laminin/metabolism , Laminin/pharmacology , Muscle Proteins/metabolism , Cells, Cultured , Endothelial Cells/metabolism , Endothelial Cells/drug effects , Endothelial Cells/cytology , Fibronectins/metabolism , Fibronectins/pharmacology , Antigens, CD/metabolism , Myocardium/metabolism , Myocardium/cytology , Stem Cell Niche/drug effects , Stem Cell Niche/physiology , Collagen Type I/metabolism , Spheroids, Cellular/drug effects , Spheroids, Cellular/metabolism , Spheroids, Cellular/cytology , Cell Culture Techniques, Three Dimensional/methodsABSTRACT
Murine peritoneal macrophages isolated from the lavage fluid after administration of thioglycolate and concanavalin A are presented by two populations of cells of different diameters. Polarization of macrophages into a proinflammatory (M1) phenotype is accompanied by an increase in number of small cells. Macrophages obtained after administration of thioglycolate demonstrate higher tendency to anti-inflammatory (M2) phenotype, while macrophages isolated after administration of concanavalin A are committed in the proinflammatory direction. Lactate level is increased in M1 macrophages in comparison with M2 cells, which indicates predominance of glycolytic metabolism. Macrophages obtained after administration of concanavalin A have reduced mitochondrial potential, which reflects a tendency to apoptosis. Autophagy activation and inhibition neutralize the differences in pro- and anti-inflammatory properties of polarized macrophages obtained after thioglycolate administration, but have less pronounced effect on macrophages obtained after administration concanavalin A. Autophagy inhibitor increases mitochondrial potential in non-polarized macrophages obtained after administration of concanavalin A. These results demonstrate divergent properties of macrophages obtained after administration of glycolate and concanavalin A due to the difference in the mechanisms of experimental peritonitis.
Subject(s)
Concanavalin A/pharmacology , Macrophage Activation/drug effects , Macrophages, Peritoneal/drug effects , Thioglycolates/pharmacology , Animals , Cell Polarity/drug effects , Disease Models, Animal , Macrophages, Peritoneal/cytology , Macrophages, Peritoneal/physiology , Male , Mice , Mice, Inbred C57BL , Peritonitis/immunology , Peritonitis/pathologyABSTRACT
Urethral pain syndrome (UPS) is characterized by the occurrence of persistent or recurrent pain in the urethra in the absence of a confirmed infection and other obvious local pathological changes. The study of its pathogenetic aspects is important first of all for understanding the causes of the disease, to prescribe effective treatment, specific recommendations for the prevention and treatment of this disease are also absent. This paper presents the advanced experience of our research group on the study of the urethral state by the in vivo cross-polarization optical coherence tomography (CP OCT) method, and also the results of the microbiota analysis in the urethral tissues. The purpose of the study is to search for the risk factors for UPS and the character of changes in the urethral tissues, using the data of: 1) concomitant pathology, 2) structural changes in the urethral wall in UPS in comparison with chronic cystitis of bacterial etiology 3) studying the microbiota of urethral tissues. MATERIALS AND METHODS: The condition of the urethra was studied in 109 patients: 55 of them with UPS (group "US"), without clinical manifestations of inflammation; 41 - with chronic inflammation of the lower urinary tract of various origins (group "Inf"); in 14 patients with stones of the upper urinary tract without pyelonephritis, the urethra was taken as the norm (group "N"). All performed a clinical minimum of studies, also cystoscopy with the study of the bladder triangle, the neck of the bladder and the urethra by the method of in vivo tissue imaging - CP OCT. The device "OCT-1300U" with wavelength of 1300 nm is used. To determine the possible role of UPS disease background, the analysis of concomitant pathology preceding the development of UPS was performed. To analyze the relationship of changes in the urethral tissues with the composition of its microbiota, a PCR study of biopsies from the proximal segment of the urethra was performed in 13 patients with UPS. RESULTS: Qualitative comparison of the thickness and character of the OCT signal of the urethral wall layers observed using CP OCT in the studied groups of patients allowed us to establish that the state of the epithelium and connective tissue structures of the mucous membrane in patients with UPS is not the norm, changes are similar to those in chronic inflammation. Changes in the character of the OCT signal were recorded in all parts of the urethra, but in the middle third they are most pronounced and most critical. In UPS, there is a brightly pronounced reorganization of the connective tissue stroma components. Pronounced fibrosis of subepithelial structures (increased signal brightness in the cross-channel compared to the norm) with their thickening was recorded in 48.2% of cases, and thinning/lack of visualization of the epithelial layer was detected in 20.5%, and in chronic inflammation 55.5% and 40.6% of cases, respectively. According to the results of PCR, only one patient had significant total bacterial contamination of the biopsy (TB=104.7). In all other cases, the total bacterial mass of the biopsies was at the level of negative control. CONCLUSIONS: In patients with UPS, the presence of several concomitant, often chronic, diseases was revealed, which may be a premorbid background and one of the risk factors for the occurrence and maintenance of UPS. Pilot PCR studies of biopsies from the proximal segment of the urethra indicate that low values of bacterial contamination in the majority of patients with UPS do not exclude the possible role of bacteria in the development of the disease in some patients. The CP OCT method used in this study is currently the only one in vivo method of visualization of the urethral mucosa, which provides real-time images of structural changes in the epithelial (atrophy or hyperplasia) and connective tissue (active or latent inflammation with cellular infiltration or fibrosis) layers of the urethra, allowing better understanding of the pathogenesis of the disease and monitoring of therapy.
Subject(s)
Urethra , Urethral Diseases , Cystoscopy , Humans , Male , Pelvic Pain , Urethra/diagnostic imaging , Urinary BladderABSTRACT
OBJECTIVE: to conduct a comparative study of the composition of the microbiota of the urethra in men with sexually transmitted infections (STIs), and healthy men. MATERIAL AND METHODS: The study included 103 men aged 18 to 45 years: 42 men with urethritis caused by STIs and 61 clinically healthy men. Identification of pathogenic and conditionally pathogenic microorganisms in scrapings from the urethra was performed by PCR in real time (test system Androflor (DNA-Technology, Moscow). RESULTS: In the analysis of the total bacterial mass, it was found that the bacterial contamination of the urethral biotope in patients with STI was significantly higher than in the group of healthy men (5.8 Lg10 and 4.7 Lg10, respectively), with the highest level of bacterial contamination was detected in patients infected with N. gonorrhoeae (6.4 Lg10). Patients with STIs had significantly lower levels of relative Staphylococcus spp., Streptococcus spp., Corynebacterium spp. and their amounts in General compared to clinically healthy men: according to ROC analysis, the best diagnostic indicator (0.93+/-0.04, p<0.001), distinguishing a group of healthy individuals from patients with STI, was the amount of Staphylococcus spp., Streptococcus spp. and Corynebacterium spp. ("Amount Of Normoflor"). In patients infected with C. trachomatis, compared with clinically healthy men, the relative number was significantly higher of Bacteroides spp. / Porphyromonas spp. / Prevotella spp., Peptostreptococcus spp. / Parvimonas spp.; in patients infected with N. gonorrhoeae - Anaerococcus spp. and in patients infected with M. genitalium - Megasphaera spp. / Veillonella spp. / Dialister spp., Anaerococcus spp., Peptostreptococcus spp. / Parvimonas spp. and Eubacterium spp. CONCLUSION: An increase in the total bacterial contamination of the urethra in STI was found, most pronounced in infection with Neisseria gonorrhoeae. The best diagnostic indicator that distinguishes normal microbiota from the microbiota of patients with STIs is the sum of Staphylococcus spp., Streptococcus spp. and Corynebacterium spp. In patients with clinical signs of an inflammatory reaction and the presence of STIs, a decrease in the normoflora in all types of STIs and an increase in obligate anaerobic bacteria - Megasphaera spp. / Veillonella spp. / Dialister spp., Bacteroides spp. / Porphyromonas spp. / Prevotella spp., Anaerococcus spp., Peptostreptococcus spp. / Parvimonas spp. and Eubacterium spp.
Subject(s)
Microbiota , Sexually Transmitted Diseases , Urethra , Urethritis , Adolescent , Adult , Chlamydia trachomatis , Humans , Male , Middle Aged , Moscow , Sexually Transmitted Diseases/metabolism , Urethritis/metabolism , Young AdultABSTRACT
Resident stem cells of the heart are denoted as heterogeneous population of immature cells, which reside in the myocardium and characterized by their ability to self-renewal and are multipotent differentiation capacity into cardiomyocyte-like and vascular like cells. CSCs were originally isolated directly by long enzymatic digestion of heart tissue and selection using stem cell markers. However, long exposure to enzymatic digestion and small myocardial sample size can affect the possibility of obtaining a significant amount of viable cells. To avoid these problems, we developed a method consisting of growing of the CPC in explant culture and subsequent immunomagnetic selection.
Subject(s)
Atrial Appendage , Cell Separation , Myocardium , Stem Cells , Antigens, Differentiation/metabolism , Atrial Appendage/cytology , Atrial Appendage/metabolism , Humans , Myocardium/cytology , Myocardium/metabolism , Stem Cells/cytology , Stem Cells/metabolismABSTRACT
The article presents the results of the proximal small intestine parietal microbiocenosis research in patients with chronic pancreatitis by polymerase chain reaction in real time. The study includes an assessment of the pharmacological correction's efficiency in this category of patients.
Subject(s)
Intestinal Mucosa/microbiology , Intestine, Small/microbiology , Pancreatitis, Chronic/drug therapy , Pancreatitis, Chronic/microbiology , Adult , Case-Control Studies , Female , Humans , Male , Middle Aged , Pancreatitis, Chronic/etiology , Prebiotics/microbiology , Severity of Illness Index , Synbiotics , Treatment OutcomeABSTRACT
We present here the results of the Analysis of HLA Population Data (AHPD) project of the 16th International HLA and Immunogenetics Workshop (16IHIW) held in Liverpool in May-June 2012. Thanks to the collaboration of 25 laboratories from 18 different countries, HLA genotypic data for 59 new population samples (either well-defined populations or donor registry samples) were gathered and 55 were analysed statistically following HLA-NET recommendations. The new data included, among others, large sets of well-defined populations from north-east Europe and West Asia, as well as many donor registry data from European countries. The Gene[rate] computer tools were combined to create a Gene[rate] computer pipeline to automatically (i) estimate allele frequencies by an expectation-maximization algorithm accommodating ambiguities, (ii) estimate heterozygosity, (iii) test for Hardy-Weinberg equilibrium (HWE), (iv) test for selective neutrality, (v) generate frequency graphs and summary statistics for each sample at each locus and (vi) plot multidimensional scaling (MDS) analyses comparing the new samples with previous IHIW data. Intrapopulation analyses show that HWE is rarely rejected, while neutrality tests often indicate a significant excess of heterozygotes compared with neutral expectations. The comparison of the 16IHIW AHPD data with data collected during previous workshops (12th-15th) shows that geography is an excellent predictor of HLA genetic differentiations for HLA-A, -B and -DRB1 loci but not for HLA-DQ, whose patterns are probably more influenced by natural selection. In Europe, HLA genetic variation clearly follows a north to south-east axis despite a low level of differentiation between European, North African and West Asian populations. Pacific populations are genetically close to Austronesian-speaking South-East Asian and Taiwanese populations, in agreement with current theories on the peopling of Oceania. Thanks to this project, HLA genetic variation is more clearly defined worldwide and better interpreted in relation to human peopling history and HLA molecular evolution.
Subject(s)
HLA-DP Antigens/genetics , HLA-DQ Antigens/genetics , HLA-DRB1 Chains/genetics , Asia , Ethnicity , Europe , Gene Frequency , Genetic Variation , Genetics, Population , Genotype , Haplotypes , Humans , Oceania , Population GroupsABSTRACT
The study is aimed to investigate the distribution of alleles of HLA-DRB1 gene in patients with early rheumatoid arthritis and healthy individuals in Russian population, and evaluate their significance as molecular genetic markers of rheumatoid arthritis predisposition and protection. The association between alleles of HLA-DRB1 genes, antibodies to cyclic citrullinated peptides and IgM rheumatoid factor was also studied. Low and high resolution HLA-DRB1 genotyping were compared. In the cohort of patients with early rheumatoid arthritis, the alleles of HLA-DRB1 gene were found to be markers of rheumatoid arthritis protection/risk, especially in the homozygous state. They determined production of antibodies to cyclic citrullinated peptides but were not associated with rheumatoid factor IgM levels. These findings support different autoimmune mechanisms of rheumatoid arthritis pathogenesis.
Subject(s)
Autoantibodies/genetics , Biomarkers/analysis , Genetic Predisposition to Disease , Immunogenetics/methods , Rheumatic Fever/immunology , Adolescent , Adult , Aged , Autoantibodies/immunology , Female , Follow-Up Studies , Genotype , Humans , Male , Middle Aged , Rheumatic Fever/genetics , Time Factors , Young AdultABSTRACT
We compared coronary angiography data from 65 patients with first myocardial infarction (fMI) and 65 patients with repetitive MI (reMI). Coronary angiographic status in both patients with fMI and reMI was characterized by predominance of multivessel lesions with stenoses localized in branches of both coronary arteries (CA). Contrary to fMI patients with reMI had more severe right CA involvement, greater number of occlusions and diffuse lesions in CA bed. Differences between angiography data between fMI and reMI were more pronounced in men than in women. Angiographic differences between fMI and reMI did not depend on the presence of history of arterial hypertension and were considerably attenuated by diabetes mellitus.
Subject(s)
Coronary Angiography/methods , Myocardial Infarction/diagnostic imaging , Female , Follow-Up Studies , Humans , Male , Middle Aged , Recurrence , Reproducibility of Results , Retrospective StudiesABSTRACT
THE AIM OF THIS STUDY: To investigate and analyse the gastrointestinal mucosal-associated microbiota in the samples from diarrhea-predominant IBS (D-IBS), constipation--predominant IBS (C-IBS) patients and healthy controls. METHODS: Oral cavity, duodenal, colonic and rectal mucosal samples were obtained from 40 IBS patients (20 C-IBS, 20 D-IBS) and 14 healthy controls. Duodenal and colonic tissue was collected during a flexible duodenoscopy and colonoscopy. Tissue samples were frozen for further molecular analysis. DNA was extracted from all frozen samples and used to identify 29 specific bacterial groups using quantitative real-time PCR (qPCR). A statistical treatment of the received data was performed. RESULTS: The dominant groups of bacteria of various microbiotops of the gastrointestinal tract in IBS patients and healthy controls were determined. qPCR analysis of duodenal samples demonstrated a reduction in the Bifidobacterium concentration in tissue samples from C-IBS patients when compared to healthy controls (p < 0.05). Analysis of rectal samples demonstrated an increase in concentrations of Faecalibacterium praustnizi (p < 0.05) and a reduction in the concentration of Streptococcus spp. (p < 0.01), Atopobium claster (p < 0.05), Ralstonia spp.+Burkholderia spp (p < 0.05) in tissue samples from D-IBS and C-IBS patients when compared to healthy controls. CONCLUSIONS: Our molecular data indicate that quantitative differences exist in specific bacterial groups in the microbiota between IBS and healthy subjects. The concentration of representatives of Bacteroidetes phylotypes appeared to be the most stable in various microbiotops of the gastrointestinal tract. Small intestinal bacterial overgrowth in IBS patients was not found. Received data help to suggest correlation with the features of the microbiota with clinical form of IBS.
Subject(s)
Bacteria , Gastric Mucosa/microbiology , Intestinal Mucosa/microbiology , Irritable Bowel Syndrome/microbiology , Adolescent , Adult , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Female , Humans , Irritable Bowel Syndrome/genetics , Male , Middle Aged , Real-Time Polymerase Chain Reaction/methodsABSTRACT
The review of studies of Russian researchers on theoretical and practical aspects of genetic predisposition to type 1 diabetes associated with immunity: HLA and not HLA genes. Most important for practical public health outcomes are evidence that HLA-genetic predisposition to type 1 diabetes is associated with the DRB1-genotype, consisting entirely of variants DRB1-genes associated with the development of T1D. It was also established that CTLA4 gene has an independent predictive value for T1D.
Subject(s)
Diabetes Mellitus, Type 1 , Genetic Predisposition to Disease , Genetic Testing/methods , HLA-DR Antigens , Immunogenetic Phenomena , Diabetes Mellitus, Type 1/diagnosis , Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/immunology , Genetic Association Studies , HLA-DR Antigens/genetics , HLA-DR Antigens/immunology , Humans , Major Histocompatibility Complex/genetics , Major Histocompatibility Complex/immunology , Pedigree , Polymorphism, Genetic/immunology , Predictive Value of TestsABSTRACT
The analysis of five Alu insertion loci (ACE, AP4OA1, B65, PV92, TPA25) has been carried out for the first time in 10 Russian populations (1088 individuals), covered all parts of historical area of the Russian ethnos. Depending on locus, Russian populations exhibit similarity with their western (European populations) or with the eastern (populations of the Ural region) neighbors. Considering frequencies of the studied Alu-insertions, Russian gene pool exhibits low variation: average difference between populations is d = 0.007, whereas on classical markers, mtDNA and Y chromosome heterogeneity of Russian gene pool is essentially higher (0.013, 0.033 and 0.142 respectively). Therefore, this set of five Alu insertions has lower variability on the intra-ethnic level. However in inter-ethnic comparisons the clear pattern was obtained: 13 Eastern European ethnic groups formed three clusters, according with their historical and geographical position--East Slavic, Caucasian and South Ural clusters. The obtained data confirms efficiency of using Alu insertions for studying genetic differentiation and history of a gene pool of the Eastern European populations.
Subject(s)
Alu Elements/genetics , Gene Pool , Genetic Loci/genetics , Mutagenesis, Insertional/genetics , Phylogeny , Apolipoprotein A-I/genetics , Female , Humans , Male , Peptidyl-Dipeptidase A/genetics , RussiaABSTRACT
In heart attack, FSTL-1 is actively secreted by cardiomyocytes, accelerates growth of heart myofibrils and stimulates of vascular endothelial growth factor expression. The aim of this work was to investigate the effect of Etoxidol on synthesis of FSTL-1 in rats after myocardial infarction. The experiments were performed on Wistar rats weighing 250-350 g with simulated myocardial infarction or intact (group 5). Animals of control groups (groups 1, 2) were treated with saline for 7 and 14 days; Ethoxidol (24 mg/kg) was injected to animals of experimental groups (group 3, 4) (the daily dose was 6.36 mg/animal) for 6 or 14 days. The injection volume was 0.2 ml. At the beginning and at the end of the study plasma concentrations of FSTL-1 were determined by the ELISA method. Myocardial FSTL-1 gene expression was determined by real-time PCR. At the end of the experiments, the hearts were also used for histochemical analysis. To determine the size of the scar formed after the modeled heart attack, we used the classic Mallory staining method. The results show that the development of experimental acute myocardial infarction is accompanied by a significant increase in FSTL-1 expression in the heart, which was detected on the 7th day and stored increased by 14 days after a heart attack. After therapy with Ethoxidol, a tendency to a decrease in the expression of FSTL-1 by the 14th day was observed; it coincided with the dynamics of the plasma protein FSTL-1 level. It can be assumed that the downregulation trend in the FSTL-1 expression is associated with a more effective repair process after a heart attack, since FSTL-1 increases precisely in response to myocardial damage and decreases when the incentives for its expression from damaged heart tissue are reduced. Indirectly, this assumption is confirmed by the detected tendency to reduce the size of post-infarction fibrosis in the treatment with Ethoxidol. The results indicate the ability of Ethoxidol to influence FSTL-1 synthesis of in rats after myocardial infarction.
Subject(s)
Cardiotonic Agents , Follistatin-Related Proteins , Follistatin , Myocardium , Animals , Cardiotonic Agents/pharmacology , Follistatin-Related Proteins/genetics , Follistatin-Related Proteins/metabolism , Myocardium/metabolism , Rats , Rats, Wistar , Vascular Endothelial Growth Factor AABSTRACT
New original data are presented on the use of achievements in human molecular immunogenetics in the management of type 1 diabetes mellitus. They include materials allowing for the prediction of the development of the disease at the population, family, and individual levels along with novel approaches to its radical treatment by the reconstitution of the lost glucose tolerance. The reported data may find wide application in current clinical practice. They open up new prospects for the enhancement of efficacy of prognosis, diagnosis, and treatment of type 1 diabetes mellitus and other autoimmune diseases.
Subject(s)
Diabetes Mellitus, Type 1/genetics , Diabetes Mellitus, Type 1/blood , Diabetes Mellitus, Type 1/diagnosis , Genetic Markers , Genetic Predisposition to Disease , Genotype , Glucose Tolerance Test , HLA Antigens/genetics , Haplotypes , Humans , Immunogenetics , Major Histocompatibility Complex , Mass Screening , Polymorphism, Genetic , Prognosis , Risk Factors , RussiaABSTRACT
The adult heart contains small populations of multipotent cardiac progenitor cells (CPC) that present a convenient and efficient resource for treatment of myocardial infarction. Several clinical studies of direct CPC delivery by injection have already been performed but showed low engraftment rate that limited beneficial effects of procedure. «Cell sheet¼ technology has been developed to facilitate longer retention of grafted cells and show new directions for cell-based therapy using this strategy. In this study we hypothesized that СPC-based cell sheet transplantation could improve regeneration after myocardial infarction. We demonstrated that c-kit+ CPC were able to form cell sheets on temperature-responsive surfaces. Cell sheet represented a well-organized structure, in which CPC survived, retained ability to proliferate, expressed progenitor cell marker Gata-4 formed connexin-43+ gap junctions, and were surrounded by significant amount of extracellular matrix proteins. Transplantation of cell sheets after myocardial infarction resulted in CPC engraftment as well as their proliferation, migration, and differentiation; cell sheets also stimulated neovascularization and cardiomyocyte proliferation in underlining myocardium and ameliorated left ventricular remodeling. Obtained data strongly supported potential use of CPC sheet transplantation for repair of damaged heart.
Subject(s)
Myocardial Infarction/therapy , Myocytes, Cardiac , Stem Cells , Vascular Remodeling , Animals , Male , Myocardium , Rats , Rats, WistarABSTRACT
Peripheral nerve injury remains a common clinical problem with no satisfactory treatment options. Numerous studies have shown that hepatocyte growth factor (HGF) exerts neurotrophic effect in motor, sensory, and parasympathetic neurons in addition to mitogenic, morphogenic, angiogenic, antiapoptotic, antifibrotic, and anti-inflammatory effect on various tissues and cells. In our study we examined efficacy of gene therapy with HGF-bearing plasmid (pC4W-hHGF) to improve consequences of traumatic nerve injury in mice. Treatment by pC4W-hHGF led to restoration of nerve structure and functional recovery compared to similar parameters in control animals. Compound action potentials (CAP) in experimental groups treated with 100 or 200⯵g of pC4W-hHGF demonstrated increased amplitude and latency decrease compared to spontaneous recovery control group. In HGF-treated mice histological analysis showed a three-fold increase in axon number in nerve portion located distal to the lesion site compared to control. Moreover, significant functional recovery of n. peroneus communis triggered by pC4W-hHGF gene therapy was observed using the footprints analysis. Obtained results provide evidence for plasmid-based HGF gene therapy as a potential treatment for traumatic injury of peripheral nerve.
Subject(s)
Genetic Therapy/methods , Hepatocyte Growth Factor/administration & dosage , Nerve Regeneration/drug effects , Peripheral Nerve Injuries/drug therapy , Plasmids/administration & dosage , Sciatic Nerve/drug effects , Animals , Hepatocyte Growth Factor/genetics , Humans , Injections, Intramuscular , Male , Mice , Mice, Inbred C57BL , Nerve Regeneration/genetics , Peripheral Nerve Injuries/genetics , Plasmids/genetics , Sciatic Nerve/injuries , Sciatic Nerve/physiologyABSTRACT
The correlation between DRB1, DQA1, and DQB1 genes of HLA class II, and the development of germ cell tumors (GCTs), as well as serological response to HERV-K proteins were investigated. Genomic DNA prepared from 99 GST patients was subjected to HLA typing by polymerase chain reaction (PCR) using the set of sequence specific primers (PCR-SSP). This set of primers made it possible to detect 14 specificities of DRB 1 locus, 12 alleles and groups of alleles of DQB 1 locus, and 8 alleles of DQA1 locus. Alongside with the definition of the occurrence of HLA markers in the total group of patients, the frequency of the occurrence of HLA-DR-DQ alleles was calculated in: 1) patients with different morphological forms of GSTs (seminomas and non-seminomas); 2) GCT patients producing or non-producing antibodies to Gag and/or Env HERV-K proteins. The comparison group consisted of 300 Moscow blood donors. The study did not reveal statistically significant differences in the frequency of the occurrence of DRB1, DQA1, and DQB1 alleles between the total group of GCT patients, its subgroup, and the control group. Thus, the data obtained demonstrated the absence of a strict correlation between the distribution of HLA class II alleles and GCT occurrence in the Russian population, as well as the ability of GCT patients to develop an antibody to HERV-K proteins, though more numerous observations are required to confirm this conclusion.
Subject(s)
HLA-DQ Antigens/genetics , Neoplasms, Germ Cell and Embryonal/ethnology , Neoplasms, Germ Cell and Embryonal/genetics , Seminoma/ethnology , Seminoma/genetics , Female , Gene Frequency , Genetic Predisposition to Disease , HLA-DQ beta-Chains , Humans , Internal-External Control , Male , Russia/epidemiologyABSTRACT
The present experiments were directed to determine the proportions of the cholinergic and GABAergic septo-hippocampal projection neurons in NMRI mice. For the labeling of the septal neurons we used a double immunocytochemical method combined with retrograde transport of wheatgerm agglutinin apo-horseradish peroxidase-gold (WAHG) injected unilaterally into the hippocampus. Monoclonal antibodies against choline acetyltransferase (ChAT) and parvalbumin (PARV) were used as markers for cholinergic and GABAergic neurons in the medial septum/diagonal band complex (MS/DB). Both antibodies were visualized in the same section using the ABC detection system with diaminobenzidine and 4-chloro-1-naphtol as chromogens. Cholinergic and PARV-containing neurons are coexisting in the MS/DB. About 38% of all retrogradely labeled neurons were ChAT-positive whereas only 10% of all retrogradely labeled cells were immunostained for PARV. On the other hand, 40% of all ChAT-positive neurons and about 24% of all PARV-positive neurons were retrogradely labeled. No double immunolabeled neurons were detected. The proportion of GABAergic neurons may have been underestimated because immunostaining for PARV only labels a subpopulation of GABAergic neurons. The present results were compared with those of previous studies in rats. They may serve as a basis of further comparative studies in mice.