Enteropathogenic Escherichia coli (EPEC) strains are a significant cause of acute and persistent diarrhea in humans, which are transmitted primarily through consumption of contaminated foods. The present study was performed for the... more
Enteropathogenic Escherichia coli (EPEC) strains are a significant cause of acute and persistent diarrhea in humans, which are transmitted primarily through consumption of contaminated foods. The present study was performed for the identification of EPEC strains from contaminated meat by a PCR technique. A panel of reference E. coli strains purchased from NBIMCC; Sofia, Bulgaria was used for the experiments. Using the optimized amplification conditions, PCR with the purified DNA template was amplified for eae gene and obtained amplicons were visualized by 2% agarose gel electrophoresis. Our result demonstrated that two strains: E. coli O 111-НБПМКК-4447 and E. coli O 26-НБПМКК-7434 have amplified a 482 bp fragment with used primers. In our experimental condition the lowest cell concentration which exhibit well visualized 482 bp amplification band is less then 54 viable E. coli cells in 1 ml. The lowest bacterial cell concentration (CFU/g) in the contaminated beef meat samples which ...
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ABSTRACT A high-performance liquid chromatography (HPLC) method with fluorescent detection was validated to analyze fluoroquinolones in small volumes of serum samples from avian species. A reversed-phase column was used. Excitation and... more
ABSTRACT A high-performance liquid chromatography (HPLC) method with fluorescent detection was validated to analyze fluoroquinolones in small volumes of serum samples from avian species. A reversed-phase column was used. Excitation and emission wavelengths for ciprofloxacin and enrofloxacin were set at 277 nm and 418 nm, respectively. These parameters for danofloxacin were 295 nm and 500 nm and for marbofloxacin were 338 nm and 425 nm, respectively. The mobile phase consisted of acetonitrile and phosphate buffer at a flow rate of 1 mL/min. The standard curves for the tested fluoroquinolones were linear over a range 10–1000 ng/mL. Enrofloxacin did not interfere with ciprofloxacin, danofloxacin, and marbofloxacin. It was used as an internal standard for danofloxacin and marbofloxacin detection. The inter-assay precision varied between 1.73 and 13.21%. Values for intra-assay precision were between 1.11 and 7.18%. Accuracy was higher than 90% at all of the tested concentrations and compounds. Mean recovery was higher than 85% for ciprofloxacin and enrofloxacin and >95% for danofloxacin and marbofloxacin.
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The induction of IL-12 and IL-10 production by the immunomodulatory C3 binding glycoprotein (C3bgp) was studied on human peripheral blood mononuclear cells (PBMC). Isolated PBMC from healthy human donors were cultured 24 hours with C3bgp... more
The induction of IL-12 and IL-10 production by the immunomodulatory C3 binding glycoprotein (C3bgp) was studied on human peripheral blood mononuclear cells (PBMC). Isolated PBMC from healthy human donors were cultured 24 hours with C3bgp isolated from Cuscuta europea. We studied also the influence of C3bgp on the cytokine production in LPS-, PHA- and PWM-stimulated PBMC. The levels of these cytokines in the various culture supernatants were subsequently determined using ELISA. Results showed that amount of IL-12 was significantly higher in C3bgp-stimulated cultures in comparison with LPS-, PHA- and PWM-stimulated PBMC. C3bgp also significantly increased IL-12 in PHA- or PWM-stimulated cultures, but not in LPS-stimulated culture. C3bgp alone was a weak inducer of IL-10 production, compared to the other stimuli. Furthermore it inhibited IL-10 production after LPS, PHA and PWM stimulation. Taken together these results indicate that immunomodulatory C3 binding glycoprotein has the abili...
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The main regulator of anti-tumor immune response is the activity of monocytes, suggesting that the produced cytokines may have a prognostic role. This study investigates gene expression of interleukin (IL)-12-related cytokine and IL-10 in... more
The main regulator of anti-tumor immune response is the activity of monocytes, suggesting that the produced cytokines may have a prognostic role. This study investigates gene expression of interleukin (IL)-12-related cytokine and IL-10 in stimulated monocytes from colorectal cancer (CRC) patients. Relative quantification of IL-12A, IL-12B, IL-23A and IL-10 mRNA transcripts was performed on the third hours after stimulation by real-time qPCR. We also explored an inhibitor of JNK signaling pathway activation for the observed cytokine gene expression. A strong downregulation of IL-12B mRNA expression in CRC monocytes compared to healthy donors was observed. The rate of transcription of IL-12B in stimulated monocytes was associated with the stage of CRC. The expression of IL-12A gene in stimulated monocytes from patients with advanced was lower than early cancer. Moreover, we observed stage dependent JNK inhibition mediated reduction in IL-12A expression. The hyporesponsiveness was strongly expressed in monocytes from advanced then early stages of CRC. Expression of IL-10 mRNA was almost equally in CRC monocytes from early stages and healthy donors. We demonstrated that altered gene expression profiles of IL-12A, IL-12B, IL-23A at mRNA level in CRC monocytes was associated with tumor development and can be attributed to anticancer immune response.
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This study investigated the functionality of the -1082(A/G) polymorphism in the promoter of the IL- 10 gene and +16974 (A/C) SNP in 3'UTR of the IL-12B gene in association with the cytokine production from stimulated PBMC in relation... more
This study investigated the functionality of the -1082(A/G) polymorphism in the promoter of the IL- 10 gene and +16974 (A/C) SNP in 3'UTR of the IL-12B gene in association with the cytokine production from stimulated PBMC in relation with sepsis. A group of 33 patients in ICU and 99 healthy volunteers were included. The SNP in IL-12B gene was detected
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The expression of many inducible genes involved in cell growth and differentiation is regulated mainly by mitogen-activated protein kinases (MAPK)-signalling pathways. In this study we examined how JNK(SP600125) and p38(SB202190) MAPKs... more
The expression of many inducible genes involved in cell growth and differentiation is regulated mainly by mitogen-activated protein kinases (MAPK)-signalling pathways. In this study we examined how JNK(SP600125) and p38(SB202190) MAPKs inhibitors influenced the inducible IL-12p40, IL- 12p70 and IL-23 production. The quantity determination of these cytokines was performed by ELISA in culture supernatants. The inhibition of both JNK and p38 increased IL-12p40 production induced by all stimuli. The inhibition of p38MAPK down-regulated IL-23 production and up-regulated IL- 12p40/p70 production in stimulated cells. We suggest the benefit of p38 control in the treatment of inflammatory and/or autoimmune diseases.
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The aim of study was to investigate the association between serum levels of IL-12p40 and IL-10 and progression of the colorectal cancer (CRC). Be observed an association between severity of CRC and serum levels of investigated cytokines.... more
The aim of study was to investigate the association between serum levels of IL-12p40 and IL-10 and progression of the colorectal cancer (CRC). Be observed an association between severity of CRC and serum levels of investigated cytokines. IL-12p40 was in the highest level in stage-I (423.6 +/- 224.7 pg/ml) compared to more severe stages. In the opposite direction were the data for immunosuppressive cytokine, IL-10. Patients with stage-IV had the highest level of IL-10 (36.02 +/- 9.5 pg/ml). On the basis of our results we could conclude that IL-12p40 and IL-10 serum levels were associated with tumour progression. More severe stages of CRC are characterized by low IL-12p40 and high IL-10 serum levels.
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The aim of the present study was to investigate the correlation between tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12p40-containing cytokines, in silicosis patients and healthy donors exposed to silica dust, in an... more
The aim of the present study was to investigate the correlation between tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12p40-containing cytokines, in silicosis patients and healthy donors exposed to silica dust, in an attempt to clarify the reason for variety of clinical outcomes between humans with similar exposure history. Serum levels of TNF-alpha, IL-12p40, IL-12p70 and IL-23 in total group of 62 silicosis patients, 24 healthy donors with similar exposure history like patients and 19 healthy donors without exposure were determined by enzyme-linked-immunosorbent assay (ELISA) method. The serum level of TNF-alpha was significantly higher in healthy donors exposed to SiO(2) (22.4 +/- 11.1 pg/mL) in comparison with non-exposed healthy donors (14.8 +/- 8.8 pg/mL; p = 0.022) and similar to that in silicosis patients. In total, group of silicosis patients significantly elevated levels of TNF-alpha (20.9 +/- 12.9 vs. 14.8 +/- 8.8 pg/mL; p = 0.047) and IL-12p40 (94.5 +/- 51...
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In this study, IL-6 and IL-12p40 production and cell viability of peripheral blood mononuclear cells from silicosis patients after in vitro stimulation were investigated. Furthermore, the effects of introducing acetylsalicylic acid to... more
In this study, IL-6 and IL-12p40 production and cell viability of peripheral blood mononuclear cells from silicosis patients after in vitro stimulation were investigated. Furthermore, the effects of introducing acetylsalicylic acid to stimulated patients' peripheral blood mononuclear cells on cytokine production and cell viability were determined. Nine patients with moderate silicosis, 11 with severe silicosis and 14 healthy subjects were recruited for this study. The level of IL-6 produced by patients peripheral blood mononuclear cells decreased depending on the stage of the disease. The addition of acetylsalicylic acid had significantly suppressive effect on the IL-6 production by lipopolysaccharide-stimulated patients' peripheral blood mononuclear cells. Acetylsalicylic acid treatment of C3 binding glycoprotein-stimulated patients' peripheral blood mononuclear cells led to significant upregulation of IL-12p40 production. Results showed a stage-dependent decrease of ce...
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To study IL-6, IL-10, IL-12 levels and circulating immune complexes (CIC) containing IgG, IgM or IgA in sera of 14 TAO patients and 12 healthy blood donors. To evaluates the ability of TAO PBMC to produce IL-6, IL-10, IL-12, as well as to... more
To study IL-6, IL-10, IL-12 levels and circulating immune complexes (CIC) containing IgG, IgM or IgA in sera of 14 TAO patients and 12 healthy blood donors. To evaluates the ability of TAO PBMC to produce IL-6, IL-10, IL-12, as well as to detect PBMC apoptosis after stimulation with different stimuli. In vitro stimulation of PBMC with lypopolysaccharide (LPS), phytohemagglutinin (PHA), C3 binding glycoprotein from uscuta europea (C3bgp), pokeweed mitogen (PWM), and dexamethasone (DM) were performed. The quantities of the secreted cytokines in sera and in culture supernatants, as well as CIC were detected by ELISA. The apoptosis was assessed according to nuclear morphology, after acridine orange staining, by fluorescence microscopy. Significantly higher IL-6 levels in the patients', than in the controls' sera was found. An increased production of IL-6 and IL-12 in TAO PBMC supernatants was detected, regardless of the stimuli used. A hyporeactivity of TAO PBMC toward IL-10 pro...
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Human macrophages and dendritic cells produce Neopterin derivates after stimulation with interferon-γ and serves as a marker of activated cell-mediated immune response. Aim: Assessment of serum neopterin levels in patients with different... more
Human macrophages and dendritic cells produce Neopterin derivates after stimulation with interferon-γ and serves as a marker of activated cell-mediated immune response. Aim: Assessment of serum neopterin levels in patients with different X-ray chances about silicosis. Methods: The serum neopterin concentrations were measured using ELISA in 60 patients with silicosis, according to conventional X-ray observation (ILO, 2002) and 16 healthy
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Research Interests: Immunology, Polymorphism, Bulgaria, Humans, Coal Mining, and 7 moreFemale, Male, Clinical Sciences, Aged, Middle Aged, Adult, and Silicosis
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Research Interests: Immunology, Methodology, Rheumatoid Arthritis, Comparative Study, Antibodies, and 19 moreHumans, Systemic Lupus Erythematosus, Systemic sclerosis, Method, P-glycoprotein, Temperature, Deposition, Glycoproteins, Antibody, Optometry and Ophthalmology, Reproducibility of Results, Correlation coefficient, Bovine Serum Albumin, Sensitivity and Specificity, Immune Complex, PLANT PROTEINS, Enzyme Linked Immunosorbent Assay, Immunoglobulin, and immunoglobulin G
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Research Interests: Genetics, Polymorphism, Bulgaria, Colorectal cancer, Humans, and 20 moreFemale, Male, Polymerase Chain Reaction, Glutathione, Statistical Significance, Detoxification, Enzyme, Clinical Sciences, Aged, Middle Aged, Genotype, Adult, Colorectal Disease, Genetic Polymorphism, Amino Acid Substitution Rates, Case Control Study, Allele Frequency, Gene Expression Regulation, Case Control Studies, and Colorectal Neoplasms
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ABSTRACT The IGF-1R signalling pathway can positively regulate cell-cycle progression and thus play a critical role in cancer development. Although recent studies provide sufficient evidence supporting the functional importance of IGF-1R... more
ABSTRACT The IGF-1R signalling pathway can positively regulate cell-cycle progression and thus play a critical role in cancer development. Although recent studies provide sufficient evidence supporting the functional importance of IGF-1R in cancer, the prognostic significance of IGF-1R expression levels to colorectal cancer (CRC) remains obscure. Expression of IGF-1R mRNA was examined in paired samples of CRC and adjacent normal mucosa, as well as in CRC patients' venous blood. We also investigated the effect of two monocytes stimulus—C3bgp and LPS on induced mRNA of IGF-1R from normal and colorectal human monocytes. The role of a member of MAPK signal transduction pathways—JNK in IGF-1R expression was assessed. The expression of IGF-1R mRNA was measured by relative RT-PCR. The results demonstrated that expression of IGF-1R mRNA in venous blood from CRC was down-regulated compared to venous blood from healthy donors (1.426 vs. 0.645; p=0.024). Mean IGF-1R mRNA level was found to be approximately 5.8 fold higher in tumour tissue compared to adjacent normal mucosa (p=0.02). Strong IGF-1R mRNA expression was found for early stages of CRC. The results showed that both stimuli used, strongly up-regulated mRNA expression for IGF-1R in CRC monocytes, then in monocytes from healthy donors. The highest level of IGF-1R expression was detected after C3bgp stimulation, regardless of JNK inhibitor presence. The significance for increasing expression of IGF-1R was observed for early CRC when patients were divided according to the tumour stage. We can conclude that downregulation of mRNA expression of IGF-1R in peripheral blood cells and stimulated monocytes from patients with advanced stages of colorectal cancer are a hallmark of tumour progression and can be used as a prognostic factor.
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... 2004). Humoral immune response to hemocyanin in inbred and outbred rabbits. ... Record number, BG2004000297. Corporate author, Trakia University, Stara Zagora. Date of publication, 2004. AGRIS Categories, Veterinary science and... more
... 2004). Humoral immune response to hemocyanin in inbred and outbred rabbits. ... Record number, BG2004000297. Corporate author, Trakia University, Stara Zagora. Date of publication, 2004. AGRIS Categories, Veterinary science and hygiene. ... Language, English. Notes, 3 ill.; 12 ...