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The Results of Fetal Chondrocytes Transplantation in Patients with Rheumatoid Arthritis
Get PDFIntroduction. Nowadays anti-inflammatory and immunosuppressive therapy has significantly improved the quality of life and prognosis of rheumatoid arthritis (RA). Nevertheless, there are still many patients with progressive rheumatoid inflammation, resulting in the destruction of joints. Cell therapy seems like a promising direction in rheumatology. The aim of our research was to evaluate the efficacy of fetal chondrocyte transplantation in patients with RA.Methods. We examined 60 patients with rheumatoid arthritis (I - III stages) between 20 and 63 years of age. They were divided into 2 groups: the first group underwent the fetal chondrocytes transplantation (n = 40), and the second was a control group who got conservative therapy (n = 20). Donor cells were taken from the chondrogenic layer of the humerus or femur heads and hip condyles of human embryos in gestation for 17-20 weeks. A suspension of fetal chondrocytes injected into affected areas of the articular surfaces under X-ray control. Cell viability was determined before the injection. Efficacy of the therapy was assessed by clinical, instrumental, and laboratory tests. This clinical trial was allowed by The Ministry of Public Health and Ethics Committee. All of our patients gave informed consent for the fetal chondrocytes transplantation.Results. Evaluation of the clinical manifestations of RA in the first group of patients showed 3.7 times decrease in pain and 1.6 times relief of synovitis. Complete reduction of contracture was observed in 82% of patients in the first group. Morphometric changes in X-ray demonstrated inhibition of the destruction in articular cartilage and surfaces of bones after transplantation of fetal chondrocytes. The dynamics of morphological changes in synovium showed 2.5 times reduction of the inflammatory reaction. Transplantation of fetal chondrocytes led to a significant reduction in ESR, CRP, fibrinogen , ?-globulin after a period of 12 months (p < 0.03). Furthermore, patients in the second group had 2.7 times higher risk of ankylosis compared to the first group. We did not observe any complications of fetal chondrocytes transplantation.Conclusions. Application of fetal chondrocytes therapy had the desired clinical effect, which was confirmed by reduction of the RA activity and decrease of cartilage and bone destruction.
Esophageal Cancer in Kazakhstan: Multi-omic Research Challenges
Get PDFIntroduction. Esophageal cancer (EC) is the sixth most common cancer in Kazakhstan, fifth leading cause of mortality among men, and ninth leading cause of mortality among women. Advances in high-throughput sequencing over the last decade have made mapping the whole genetic variation in genome-wide scale possible. Transcriptome sequencing has become a powerful method for detecting driver mutations in cancer, since somatic point mutations as well as aberrant RNA variants, such as fusion genes and alternative splicing, can be identified. The aim of the study was to identify the genetic basis of EC by performing whole transcriptome sequencing (RNA-Seq) study in Kazakhstani patients.Materials and methods. We included patients with EC who had been admitted to the oncology center in Astana, Kazakhstan during the 2013-2014 year period. A pair of fresh frozen EC, its adjacent normal tissue specimen, and venous blood were obtained. So far, five pairs of EC samples were subjected to RNA-seq. Total RNA was isolated, and its quality was assessed using Agilent Bioanalyzer. The cDNA library was prepared following the standard mRNA protocol by Illumina and sequenced using Illumina HiSeq2000. Bionformatic analysis is ongoing.Results. During 2013, a total of 74 patients with EC were hospitalized in the oncology center, Astana, Kazakhstan. Radical and palliative surgery was performed on 39 and 34 patients, respectively, and 1 patient refused surgery treatment. The median age of the patients was 66 years (range 49-86 years). 88.4% of the patients were diagnosed with advanced stages T3-T4, and 74.5% from them has dysphagia III-IV levels. 83% of the cases were squamous cell carcinoma (ESCC). The major localizations for this type of cancer were the middle section (58.2%), lower section (37.2%), and upper section (4.6%) of the cases.Conclusion. ESCC is the most common histologic subtype of esophageal cancer in our patients and is characterized by a poor prognosis. Most patients were diagnosed with late stages T3-T4. Using high throughput sequencing approach, we could potentially identify a higher number of crucial molecular pathways involved in esophageal carcinogenesis that could facilitate the development of new diagnostic and treatment strategies. The early detection of EC gives hope of a long-term survival for patients
Features of Cytokine Profile in Different Age Groups
Get PDFIntroduction. The study of the cytokine profile during aging is interesting because age-related changes of the immune status are usually correlate with the onset of specific diseases. Characteristics of cytokine activity in the elderly can not only detail the pathogenesis of the disease but also help to choose the appropriate therapeutic strategy, which in addition to the therapeutic effect could improve the quality of life of the elderly. The purpose of this study was to examine cytokine levels in older adults.Material and methods. We examined 268 people aged 45-80 years and older. All surveyed individuals were divided into 8 different age groups. All participants were tested for concentrations of IL-1?, IL-2, TNF-a and IFN-?.Results. The study found that concentrations of TNF-a increased with age. For age group 45-49, the concentration of TNF-a was 5.94 pcg/ml. In older age groups, there was a gradual increase in cytokine concentration. In a group of centenarians, concentration of TNF-a reached 20.55 pcg/ml, which is 3.4 times higher compared to the middle age group. Similar trends were found in the concentration of IL-1. For the age group of 45-49, the concentration of IL-1 b was 3.38 pcg/ml, and in the age group of 80 years and older, levels of this cytokine increased almost 5 times. It was found that with age-related there is a gradual decrease in the level of IL-2, and a gradual increase of IFN-?. The decrease in IL-2 is due to the typical aging decrease in the amount of T-lymphocytes.Conclusion. Thus, our results indicate that there are significant deviations of immune parameters, particularly in cytokine concentrations, in older adults compared to middle aged adults.
The Probiotical Potential of Lactobacilli from Therapeutic Preventive Beverage Kurunga
Get PDFIntroduction. Kurunga is a dairy drink made of a mix of lactic acid and alcoholic fermentation, characterized by high biological value based on protein composition, amino acid spectrum, fatty acid composition of lipids, vitamin and mineral substances, and physiological activity of microbiota containing lactobacilli, lactococci, bifidobacteria, and yeast. Among the probiotic correctors of normal microbiota isolated from national products, lactobacilli was of particular interest, with regards to a therapeutic – preventive effect. The aim of the study was to examine the probiotic properties of lactobacilli from kurunga.Methods. We isolated lactic acid bacteria strains from kurunga. The isolated cultures were identified using common microbiological methods and phylogenetic analysis. The antibiotic activities of these strains were determined by measuring the growth inhibition zone of test cultures. The probiotic properties were measured as levels of resistance to bile and hydrochloric acids, in addition to the presence of superoxide dismutase (SOD) activity using the xanthine oxidase-cytochrome method. Proteolitic activity was determined at the various levels of pH (3.0, 4.2, 5.3, and 7.0).Results. According to the morphological, cultural, physiological, biochemical properties and the genotypic analysis of the oligonucleotides sequence of specific genes, the most effective strain was identified as Lactobacillus diolivorans KL-2 (GenBank database KC438372). The isolated strain suppressed the growth of Gram-positive bacteria, such as Bacillus, Staphylococcus, and Listeria sp., as well as Gram-negative bacteria, such as E.coli, Proteus, Salmonella sp. They also possessed fungicidal action (based on Penicillum, Aspergillus sp, and Candida sp.). The strain was resistant to the action of the bile acids at concentrations of 0.8% to 1.0% and hydrochloric acid. The strain KL-2 possessed a relatively high SOD activity (25.74 U/mg of protein), a low proteolytic activity at a pH 3.0 (4.74·10-3 PU/ml), and high proteolytic activity at pH 4.2 (294.74·10-3 PU/ml), pH 5.3 (330.52·10-3 PU/ml) and pH 7.0 (713.68·10-3 PU/ml).Conclusion. The unique properties of this strain, such as stability in the gastrointestinal tract, the wide spectrum of bactericidal and fungicidal action to the pathogenic species, the relatively high superoxide dismutase and proteolytic activities, and the absence of toxicity, make it a prime candidate for probiotic culturing
Role of ROS in A?42 Mediated Activation of Cerebral Endothelial Cells
Get PDFIntroduction. There is substantial evidence that the deposition of aggregated amyloid-beta peptide (A?) in brain parenchyma and brain vessels is the main cause of neuronal dysfunction and death in Alzheimer’s disease (AD). A? exhibits multiple cytotoxic effects on neurons and glial cells and causes dysfunction of the blood brain barrier (BBB). In AD brains, an increased deposition of A? in the cerebral vasculature has been found to be correlated with increased transmigration of blood-borne inflammatory cells and neurovascular inflammation. However, regulatory mediators of these processes remain to be elucidated. In this study, we examined the role of ROS in actin polymerization and expression of adhesion molecules (P-selectin) on the surface of the cerebral endothelial cells (CECs) that are activated by A?42.Materials and methods. Mouse BEnd3 line (ATCC) was used in this research. BEnd3 cells respond to A? treatment similarly to human primary CECs and are a common model to investigate CECs’ function. We used immortalized bEnd3 cells as the following: controls; cells incubated with A?42 for 10, 30, and 60 minutes; cells incubated with 30 mM of antioxidant N-acetylcysteine (NAC) for 1 hr; and, cells pre-treated with NAC followed by A?42 exposure. We measured DHE fluorescence to investigate intracellular ROS production. Immunofluorescent microscopy of anti-P-selectin and oregon green phalloidin was used to quantify the surface P-selectin expression and actin polymerization, and Western blot analysis was used to analyze total P-selectin expression.Results. The results of this study have demonstrated a significant time-dependent ROS accumulation after 10 minutes, 30 minutes, and 60 minutes of A?42 treatment, while A?42 stimulated ROS production in CECs was attenuated by pre-treatment with the NAC antioxidant. We also found that A?42 increased P-selectin fluorescence at the surface of bEnd3 cells in a time dependent manner in parallel to ROS elevation. However, total expression levels of P-selectin were not changed following exposure to A?42. Pre-treatment with NAC attenuated A?42 induced P-selectin localization, while NAC alone did not significantly affect P selectin localization. As a positive control, H2O2 also increased P-selectin expression on the cell surface, which peaked after 30 minutes of H2O2 treatment. Exposure of CECs with A?42 promoted actin polymerization, which peaked after 10 minutes of A?42 treatment, while no significant increase of F-actin intensity was observed when cells were pre-treated with NAC. H2O2 was able to mimic A?42 induced oxidative stress, causing increased actin polymerization with similar timing.Conclusions. The results of our study have indicated that A?42 induced accumulation of P-selectin on the surface of bEnd3 cells and promoted actin polymerization, and all these events were correlated with ROS generation. The rapid post-translational cell signaling response mediated by ROS may well represent an important physiological trigger of the microvascular inflammatory responses in AD and requires further investigations
Lactobacillus for Vaginal Microflora Correction
Get PDFIntroduction. Despite the significant progress made in prevention, diagnosis, and treatment, there is still a high rate of vaginal dysbiosis in Kazakh women. The use of antibiotics in the treatment of vaginal dysbiosis contributes to the elimination of pathogens as well as microflora, which can lead to a decrease in local immunity and more favorable conditions for infection spread. The most physiologically safe and promising method for the restoration of vaginal biocenosis is the use of probiotics administered by a vaginal route.Methods. We have allocated 64 of cultures of Lactobacillus from the vaginal epithelium of healthy women of reproductive age and women with diagnosed bacterial vaginosis (BV). Identification of cultures was performed by PCR analysis of 16S ribosomal RNA. Evaluation of biological significance was determined by the following criteria: high antagonistic activity against Candida albicans, Escherichia coli, Serratia marcescens, Proteus mirabilis, Klebsiella ozaenae, and Staphylococcus aureus; and production of hydrogen peroxide, resistance to antibiotics, adhesive activity. We studied the symbiotic relationship of selected biologically active of cultures to each other and received options for consortiums with properties of probiotics through co-cultivation.Results. Results of genotyping showed that the isolated lactobacilli belong to the seven species: L. fermentum, L. salivarius, L. gasseri, L. crispatus, L. jensenii, L. plantarum, and L. delbrueskii. L. fermentum, L. salivarius, L. gasseri, and L. jensenii occur in women with suspected BV. The highest percentage of occurrence in the vagina of healthy women was L. fermentum (28%). Most strains of lactobacilli possess high inhibitory activity for all test-strains, except Candida albicans (37.5%). 56% of studied cultures revealed high adhesion to human erythrocytes. All lactobacillus strains were resistant to metronidazole, 80% to kanamycin, 57% to vancomycin, and sensitivity to roxithromycin, amoxiclav, ampicillin was diagnosed in all strains. 50% of cultures showed a moderate sensitivity to gentamicin and cefazolin. In a study of peroxide-producing activity, 80% of the cultures exhibited peroxide-producing activity. As a result of s?reening, the 7 most active strains of lactobacilli were selected for development of 10 variants of probiotic consortia. Also, there was increase of adhesive activity in the consortia compared to other components. These consortia can be used for the treatment of BV in addition to metronidazole.Conclusion. The probiotic consortia identified in this study had high antagonistic, adhesive properties, and resistance to metronidazole. These probiotics can potentially be used for the development of biological products for the treatment and prevention of bacterial vaginosis
Deprivation of human natural killer cells and antitumor immune response
Get PDFIntroduction: Cell-based immunotherapy has been given increased attention as a treatment for cancer. Human natural killer (NK) cells are resident lymphocyte populations. They exhibit potent antitumor activity without human leukocyte antigen matching and without prior antigen exposure. They also are a promising tool for immunotherapy of solid and hematologic cancers. However, most cancer patients do not have enough NK cells to induce an effective antitumor immune response. This demonstrates a need for a source of NK cells that can supplement the endogenous cell population.Material and methods: In this study, we derived induced pluripotent stem cells (iPSCs) from peripheral blood T-lymphocytes using Sendai virus vectors.Results: Generated iPSCs exhibited monoclonal T cell receptors (TCR) rearrangement in their genome, a hallmark of mature terminally differentiated T cells. These iPSCs were differentiated into NK cells using a two-stage coculture system: iPSCs into hematopoietic CD34+ cells with feeder cells M210-B4 (ATCC, USA) and CD34+ cells into mature NK cells with AFT024 cells (ATCC, USA). Our results showed that iPSC-derived NK cells expressed CD56, CD16, NKp 44 and NKp 46, possessed high cytotoxic activity and produced high level of interferon-?.Conclusion: Based on our data, derivation of NK cells from induced pluripotent stem cells should be considered in the treatment of oncologic diseases.This would allow for the development of cell therapy for cancer using immunologically compatible NK cells derived from iPSCs. This may contribute to a more efficient treatment of oncologic diseases in addition to traditional cancer treatment
Radiation risk factors in incidence anortality among exposed individuals of East Kazakhstand m
Get PDFIntroduction: Lengthy clinical and epidemiological studies at the Research Institute of Radiation Medicine and Ecology have discovered basic patterns of long-term effects from ionizing radiation in population groups exposed to radiation risk. Methodology for calculating injury from radiation risk factors has been developed and implemented to minimize the effects of the Semipalatinsk nuclear test site (SNTS).Material and methods: We analyzed materials from the database of the Scientific Medical Register that were exposed to radiation as a result of SNTS. We analyzed both male and female populations of the Abay, Beskaragai and Zhanasemei, Kokpekti (control) areas of East-Kazakhstan region (EKR) from 2008-2012. These populations were split into three groups allocated by the generation. The first group represented persons born from the period of 01/01/1930 -08/01/1949 and their children born from the period of 10/09/1949-12/31/1962. The second group were persons born after 01/01/1963. The third group served as the control and were persons who immigrated to these areas after 1990.Results: There was an increased incidence of cancer (21.5%, p < 0.000734), cardiovascular diseases (10.2%); respiratory problems (9.6%), gastrointestinal issues (9.1%, p < 0.00371-0.00679) in the first group. The effect of the radiation dose has not been fully stuided among the subjects in the second group.The major causes of excess mortality in the first group were neoplams (30.6%), hypertension (23.8%), and myocardial infarction (22.6%). The effects of radiation influenced mortality in the second group were 2-2.5 times lower than the first group.Conclusion: There is a correlation between the size of the radiation dose, the risk profile, and age at the moment of radiation exposure with trends of morbidity and mortality in the radiation exposed areas
Randomized Clinical Trial: Efficacy of a New Synbiotic in Adults with Metabolic Syndrome
Get PDFIntroduction: Metabolic syndrome is a lifestyle disease and is a frequent problem among the adult population. Human gut microbiota plays a key role in the development of metabolic syndrome. Recently, the gut microbiota has emerged as an important contributor to the development of obesity and metabolic disorders through its interactions with environmental (e.g. diet) and genetic factors. The aim of this study was to research the effects of synbiotic on the gut microbiota and host metabolism. Methods: We conducted a double-blind, randomized, placebo-controlled trial. Our sample included 180 adults (ages 30-89) with symptoms of metabolic syndrome, who were allocated to either placebo or synbiotic group. The main inclusion criteria were: blood pressure of around 130/90 mmHg; raised fasting plasma glucose (FPG) >100 mg/dL (5.6 mmol/L), previous diagnosis of type 2 diabetes, dyslipidemia triglycerides (TG) of 1.70 mmol/L, a high-density lipoprotein cholesterol (HDL-C) of 0.90 mmol/L in males and 1.0 mmol/L in females, and central obesity with a waist/hip ratio > 0.90 in males or > 0.85 in females or a body mass index > 30 kg/m2. Results: We enrolled 90 adults in the placebo group and 90 in the synbiotic group. The two groups had similar demographic and clinical characteristics. Consent was signed by all patients. All patients underwent clinical and laboratory evaluation, including complete blood tests, glucose test, glycosylated hemoglobin, total cholesterol and triglycerides, cholesterol, LDL, HDL plasma, immunogram, and coprogram. All patients were interviewed with a questionnaire that included 200 questions related to diet, lifestyle, and health. Synbiotic were used by patients in a dose of 200 grams twice a day. The duration of applying of the synbiotic was 90 days. To study the composition of the intestinal microbiota, stool samples were collected before and after applying the synbiotic. The microbial composition will be determined by analyzing the locus of 16S rDNA.Conclusion: This ongoing study is currently undergoing microbial composition analysis in order to establish the efficacy of the new synbiotic in adults with metabolic syndrome.
Complete Genome Sequence of the Probiotic Lactic Acid Bacterium Lactobacillus Rhamnosus
Get PDFIntroduction: Lactobacilli are a bacteria commonly found in the gastrointestinal tract. Some species of this genus have probiotic properties. The most common of these is Lactobacillus rhamnosus, a microoganism, generally regarded as safe (GRAS). It is also a homofermentative L-(+)-lactic acid producer. The genus Lactobacillus is characterized by an extraordinary degree of the phenotypic and genotypic diversity. However, the studies of the genus were conducted mostly with the unequally distributed, non-random choice of species for sequencing; thus, there is only one representative genome from the Lactobacillus rhamnosus clade available to date. The aim of this study was to characterize the genome sequencing of selected strains of Lactobacilli.Methods: 109 samples were isolated from national domestic dairy products in the laboratory of Center for life sciences. After screaning isolates for probiotic properties, a highly active Lactobacillus spp strain was chosen.Genomic DNA was extracted according to the manufacturing protocol (Wizard® Genomic DNA Purification Kit). The Lactobacillus rhamnosus strain was identified as the highly active Lactobacillus strain accoridng to its morphological, cultural, physiological, and biochemical properties, and a genotypic analysis.Results: The genome of Lactobacillus rhamnosus was sequenced using the Roche 454 GS FLX (454 GS FLX) platforms. The initial draft assembly was prepared from 14 large contigs (20 all contigs) by the Newbler gsAssembler 2.3 (454 Life Sciences, Branford, CT).Conclusion: A full genome-sequencing of selected strains of lactic acid bacteria was made during the study