Ludmiła Weglarz

Medical University of Silesia in Katowice, Catowice, Silesian Voivodeship, Poland

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Publications (55)51.13 Total impact

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    ABSTRACT: Inflammatory bowel disease (IBD) is chronic inflammatory condition associated with increased risk of developing colorectal cancer. A number of mediators of inflammation, such as pro-inflammatory cytokines, prostaglandins and nitric oxide have been involved in carcinogenesis, especially in the promotion and progression stages. NO is synthesized from L-arginine by constitutively expressed endothelial and neuronal nitric oxide synthases (eNOS and nNOS, respectively) and an inducible NOS (iNOS) isoform expressed under inflammatory conditions. A selective inhibitors of iNOS could be, therefore, considered to be good candidates as chemopreventive agents against colon cancer. In this study, the effect of inositol hexaphosphate (IP6), dietary phytochemical, on the mRNA expression of iNOS stimulated with bacterial lipopolysaccharides (Escherichia coli and Salmonella typhimurium) and IL-1β in intestinal cells Caco-2 for 6 and 12 h was investigated. A transcription level of iNOS with the use real time QRT-PCR technique was determined in cells treated with 1 and 2.5 mM IP6. Stimulation of Caco-2 with pro-inflammatory factors (LPS and IL-1β) resulted in an up-expression of iNOS mRNA at 6 and 12 h. Cells exposed to IP6 only revealed significant reduction in iNOS gene transcription after 12 h. A decrease in iNOS transcription by IP6 following the gene induction by proinflammatory agents in 6 and 12 h lasting cultures was also determined. The findings of this study suggest that one of the anticancer and anti-inflammatory abilities of IP6 can be realized by suppressing the expression of gene encoding inducible nitric oxide synthase isoform at the transcriptional level.
    No preview · Article · Jul 2015 · Acta poloniae pharmaceutica
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    ABSTRACT: Phytic acid (IP6) is a major fiber-associated component of a diet physiologically present in human intestines. Studies showed that this phytochemical can modulate immune functions of intestinal epithelium through regulation of proinflammatory cytokines secretion but mechanisms underlying these cellular response to IP6 have weakly been examined, as yet. The aim of this study was to determine the role of protein tyrosine kinase (PTK) in secretion of IL-8, a central proinflammatory cytokine, by unstimulated and IL-1beta-stimulated intestinal epithelial cells Caco-2 treated with IP6 (1 and 2.5 mM). To study the involvement of PTK signal pathway in IL-8 secretion, inhibitors of phosphotyrosine phosphatase (sodium orthovanadate, OV) and tyrosine kinase (genistein, GEN) were incubated with Caco-2 cells prior to IP6 treatment. IP6 had suppressive effect on basal and IL-1beta-stimulated IL-8 secretion by cells. The effect of OV on IL-8 release by cells treated with IP6 was different under constitutive and stimulated conditions. Secretion of IL-8 was significantly down-regulated in cells with GEN and GEN plus IP6 treatment. In addition, total PTK activity in both unstimulated and IL-1beta stimulated cells was determined in the presence of IP6. The results suggest that physiological intestinal concentrations of IP6 may have an inhibitory effect on IL-8 secretion by Caco-2 cells and one of the mechanisms of its action is the inhibition of PTK signaling cascade. The study revealed for the first time that PTKs could be one of the molecular targets for IP6 effects in the intestinal epithelial cells.
    No preview · Article · Apr 2013 · Acta poloniae pharmaceutica
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    ABSTRACT: Inositol hexaphosphate (IP6) is a naturally occurring phytochemical, found in abundance in cereals, legumes and other high-fiber-content diets. IP6 has shown promising efficacy against a wide range of cancers. Its anti-cancer activity involves anti-proliferative, pro-apoptotic and anti-metastatic effects. Both matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs), are implicated in tumor growth, metastasis, and angiogenesis. Phorbol-12-myristate 13-acetate (PMA) is a well-known inflammatory stimulator and tumor promoter that activates PKC and increases the invasiveness of various types of cancer cells by activating MMPs. The aim of the present study was to examine the influence of IP6 on the expression of selected MMPs, i.e., MMP-1, -2, -3, -9, 10, -13 and their TIMP-1 and -2 in unstimulated and PMA-stimulated colon cancer cell line Caco-2. Quantification of genes expression in Caco-2 cells treated with 100 ng/mL of PMA, 2.5 mM of IP6 and both for 6 and 12 h was carried out using real time QRT-PCR technique. Stimulation of cells with PMA resulted in an up-expression of MMP-2, MMP-3, MMP-9, MMP-10, MMP-13 and TIMP-1 mRNAs and decrease in MMP-1 gene expression. The quantity of TIMP-2 transcript was reduced by PMA. A significant decrease in MMP-2, MMP-3, MMP-10, MMP-13, and TIMP-1 expression in response to IP6 was observed. IP6 down-regulated MMP-9 transcription induced by PMA and decreased the level of both MMP-2 and MMP-3 mRNAs in PMA-stimulated cells. Caco-2 treated with both PMA and IP6 showed a significant decrease in MMP-1 expression in comparison to PMA-stimulated cells. The results of this study show that PMA can modulate MMP and TIMP genes transcription in colon cancer cells Caco-2. IP6 exerts an influence of basal mRNA expression of some MMPs and their tissue inhibitors and down-regulates MMP-1, MMP-2, MMP-3 and MMP-9 in cells treated with PMA. IP6 could be an effective anti-metastatic agent that suppresses expression of MMP genes at transcription level.
    No preview · Article · Jan 2013 · Acta poloniae pharmaceutica
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    ABSTRACT: Soluble adhesion molecules such as soluble intercellular adhesion molecules-1 (sICAM-1) and soluble E-cadherin (sE-cadherin) play important role in tumor invasion and the development of metastasis. It was observed that their concentrations in body fluids of patients with colon cancer were elevated. Celecoxib, a selective inhibitor of cyclooxygenase-2 (COX-2) besides its analgesic, anti-inflammatory, and antipyretic activity is able to inhibit development of colon cancer and reduce risk of metastasis. The additional factors, e.g., dietary components in colon cancer, may influence therapeutic effect of drugs, such as cytokines. TNF-alpha (tumor necrosis factor - alpha) is a cytokine, which concentration significantly increases in serum of patients with inflammatory and cancer diseases. The latest studies demonstrate, that phytic acid (IP6), a myo-inositol derivative, abundantly present in high-fiber diets could substantially reduce colon cancer incidence. The aim of the present study was to evaluate the influence of celecoxib on sICAM-1 and sE-cadherin concentrations in transformed epithelial colon cell cultures simultaneously exposed to IP6 and TNF-alpha. Additionally, the adhesion of the exposed cells to collagen I was assessed. HT-29 and Caco-2 cells were cultured in the presence of 50 ng/mL celecoxib, 1.0 mM IP6, and 100 ng/mL TNF-alpha, and their combination: TNF-alpha plus IP6, TNF-alpha plus celecoxib, IP6 plus celecoxib, and TNF-alpha with celecoxib plus IP6, for 96 h. Nonexposed cell line cultures served as controls. Concentrations of sICAM-1 and sE-cadherin were measured in the culture medium by enzyme-linked immunosorbent assay (ELISA) using Quantikine - Human sICAM-1/CD54 Immunoassay and Quantikine-Human sE-Cadherin Immunoassay. All the results obtained were expressed as ng per mL. In the adhesion assay, the cells were incubated with IP6 (0.5, 1.0 and 2.0 mM), TNF-alpha (100 ng/mL), celecoxib (50 ng/mL) and their combination for 90 min. Fluorescence values 480 nm/530 nm reflected concentrations of DNA in cells attached to collagen I. The obtained results indicate that celecoxib (50 ng/mL), the selective COX-2 inhibitor, reduces significantly sICAM-1 and sE-cadherin concentrations in HT-29 and Caco-2 transformed human epithelial colorectal cell line cultures co-treated with IP6 (1.0 mM) and TNF-alpha (100 ng/mL). A decrease of cells adhesion property to collagen I was observed under the influence of 50 ng/mL celecoxib on cell cultures exposed to 1.0 or 2.0 mM IP6 and 1.0 or 2.0 mM IP6 plus 100 ng/mL TNF-alpha.
    No preview · Article · Jan 2013 · Acta poloniae pharmaceutica
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    ABSTRACT: Transforming growth factors-beta (TGF-beta) are multifunctional cytokines involved in the regulation of cell development, differentiation, survival and apoptosis. They are also potent anticancer agents that inhibit uncontrolled proliferation of cells. Incorrect TGF-beta regulation has been implicated in the pathogenesis of many diseases including inflammation and cancer. In humans, the TGF-beta family consists of three members (TGF-beta1, 2, 3) that show high similarity and homology. TGF-betas exert biological activities on various cell types including neoplastic cells via their specific receptors. Inositol hexaphosphate (phytic acid, IP6), a phytochemical has been reported to possess various health benefits. The aim of this study was to examine the effect of IP6 on the expression of genes encoding TGF-beta1, TGF-beta2, TGF-beta3 isoforms and their receptors TbetaRI, TbetaRII, TbetaRIII in human colorectal cancer cell line Caco-2. The cells were treated with 0.5, 1 and 2.5 mM IP6 for 3, 6 and 12 h. The untreated Caco-2 cells were used as the control. Quantification of genes expression was performed by real time QRT-PCR technique with a SYBR Green I chemistry. The experimental data revealed that the TGF-beta1 mRNA was the predominant isoform in Caco-2 cells and that IP6 enhanced transcriptional activity of genes of all three TGF-beta isoforms and their receptors TbetaRI, TbetaRII TbetaRIII in these cells. At concentrations up to 1 mM, IP6 over-expressed the genes in 6 h lasting cultures, and its higher dose (2.5 mM) caused successively increasing transcript level of TGF-beta isoforms and receptors with the duration of experiment up to 12 h. The findings of this study indicate that one of anti-cancer abilities of IP6 can be realized by enhancing the gene expression of TGF-beta isoforms and their receptors at the transcriptional level.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: Phytic acid (IP6) is an essential component of high fiber diet physiologically present in human large gut. It has been recognized to possess various significant health benefits effects including chemopreventive and have antineoplastic activity against various types of cancer. Moreover, its role in immune response through modulation of the secretion of proinflammatory cytokines and chemokines has been postulated. One of the signal transduction pathways involved in a variety of inflammatory responses is p38 mitogen-activated protein kinase (MAPK) pathway. The aim of this study was to examine effect of IP6 on human p38alpha MAP kinase activity and the expression of gene encoding p38 MAP kinase in unstimulated and IL-1beta-stimulated Caco-2 cells. Furthermore, the role of signaling pathways involving p38 MAP kinase in IP6-induced down-regulation of IL-8 secretion by unstimulated and IL-1beta-stimulated cells in the presence of p38 MAP kinase activator (anisomycin) and inhibitor (SB 203580) was evaluated. IP6 inhibited activity of recombinant p38 MAPK activity in dose-dependent manner. Treatment of cells with IP6 for 3 h resulted in decreased p38 MAP kinase expression in both unstimulated and stimulated with IL-1beta cells. The similar level of p38alpha mRNA was found in untreated and treated with IP6 cells after 6 and 12 h. Incubation of Caco-2 cells with anisomycin resulted in upregulation of IL-8 secretion and their pretreatment with anisomycin prior to IP6 addition showed down-regulation of IL-8 secretion compared to cells treated with anisomycin alone. The findings of this study show that p38 MAPK could be one of the molecular targets for IP6 in the intestinal epithelial cells and that IP6 inhibitory effect on IL-8 secretion by Caco-2 cells could be mediated by its inhibition of p38 activity.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: The latest studies suggest that adhesion molecules are involved in the arising of malignant changes and in distant metastasis induction. The soluble forms of several adhesion molecules, have recently emerged as novel and potentially useful tumor markers. Among a number of identified, high interest wake soluble molecules similar to the immunoglobulin -- soluble intercellular adhesion molecules-1 (sICAM-1) and soluble E-cadherin (sE-cadherin). In the present work, the authors concentrate on one tumor type, colorectal carcinoma, in which distant metastases, are the main cause of failure, in spite of surgical curing of the primary tumor. It is known that TNF-alpha (tumor necrosis factor - alpha) serum concentration of patients with cancer is raised. The changes in soluble adhesion molecules concentrations in serum and others fluids, could be modulated by many different factors affecting cancer cells. In the case of colon cancer one of the factors is a high-fiber diet, containing an anti-cancer chemical, inositol hexaphosphate (IP6). The aim of this study was to estimate the influence of TNF-alpha on the concentration of sICAM-1 and sE-cadherin in the microenvironment of HT-29 malignant epithelial colorectal cells stimulated with IP6. Additonally, adhesive property of HT-29 human colorectal cancer cell line to collagen I was estimated. The HT-29 cells were treated with TNF-alpha (10 ng/mL and 100 ng/mL - estimation of sICAM and sE-cadherin concentration; 100 ng/mL - adhesion assay), IP6 (0.5 mM, 1.0 mM, 2.0 mM) and TNF-alpha in combination with IP6. The level of sICAM-1 and sE-cadherin in cultures of HT-29 cells was measured by enzyme-linked immunosorbent assay (R&D Systems), and adhesion of the cells to collagen I was investigated by Cyquant Proliferation Assay Kit. The present findings demonstrate that TNF-alpha and inositol hexaphosphate have an effect on the sICAM-1 and sE-cadherin concentration in cultures of HT-29 cells. IP6 at a concentration of 2.0 mM induced a decrease of sE-cadherin concentration in cultures of these cells and significantly reduced their adhesion to collagen I. TNF-alpha at concentration of 100 ng/mL caused the significant increase in the sICAM-1 level, but to a lesser degree in the presence of higher concentrations of IP6. However, TNF-alpha did not cause such a significant increase in sE-cadherin level. The sE-cadherin concentration was most likely associated with inositol hexaphosphate activity.
    No preview · Article · Nov 2012 · Acta poloniae pharmaceutica
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    ABSTRACT: We determined retrospective analysis of the diagnostic value of virus serology in patients with non-ischemic systolic heart failure and parvovirus B19 infection. Virus serology and endomyocardial biopsy were performed in 31 patients with non-ischemic systolic heart failure hospitalized from 2001 to 2006 in our clinic. The serum specimens from 31 patients were tested for IgM and IgG antibody against parvovirus B19. IgM antibodies were identified in 3 patients and IgG antibodies were identified in 23 patients. All of the patients underwent endomyocardial biopsy which revealed chronic active myocarditis in 10 patients (32.4%), chronic persistent myocarditis in 14 patients (45.1%) and no myocarditis in 7 patients (22.5%). Virus serology has no relevance for the diagnosis of non-ischemic systolic heart failure caused by parvovirus B19 infection. The result of serological tests are positive more frequently than the biopsy specimens results.
    No preview · Article · Jan 2012 · Wiadomości lekarskie (Warsaw, Poland: 1960)
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    ABSTRACT: Intestinal epithelial cells play an important role in the mucosal immune and inflammatory reactions via the expression and secretion of proinflammatory cytokines such as interleukin-6 (IL-6) and interleukin-8 (IL-8). The expression of both interleukins is regulated by nuclear factor KB (NF-kappaB). Phytic acid (IP6) is an essential component of high fiber diet. It is physiologically present in the human large gut at concentrations reaching 4 mM. It exhibits pleiotropic health beneficial effects including anti-oxidant and anti-tumor activities. Recent studies showed that IP6 can modulate immune functions of intestinal epithelium through regulation of proinflammatory cytokines secretion. The aim of this study was to analyze the effect of IP6 on the expression of IL-6 and IL-8 as well as p50 and p65 subunits of NF-kappaB and its inhibitor IkappaBalpha in Caco-2 cells stimulated with IL-1beta. A kinetic study of mRNAs expression in cells was performed after their treatment with 1 and 2.5 mM IP6 for 3, 6 and 12 h. Quantification of the genes expression was carried out using real time QRT-PCR technique. IP6 at all used concentrations had no influence on transcription of p65 gene and modulated expression of p50 and IkappaBalpha genes in Caco-2 cells. Treatment of cells with IP6 resulted in a marked decrease in both IL-6 (at 3 and 6 h) and IL-8 expression (3 h). The results of these studies suggest that IP6 may exert immunoregulatory effects on intestinal epithelium by influencing transcriptional activity of genes encoding p50 subunit of NF-kappaB, its inhibitor IkappaBalpha and proinflammatory cytokines IL-6 and IL-8.
    No preview · Article · Nov 2011 · Acta poloniae pharmaceutica
  • J. Lodowska · M. Lach · L. Weglarz
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    ABSTRACT: Retinal pigment epithelium (RPE) is among others responsible for maintenance of the visual cycle through the metabolism of vitamin A derivatives, phagocytosis and degradation of the photoreceptor outer regions and retina protection against the oxidative stress. Postmitotic RPE cells become dysfunctional with age as a consequence of metabolic and phagocytic insufficiency. Their dysfunction leads to accumulation of lipofuscin, which is considered as a biomarker of aging. Photocytotoxic components of this autofluorescent pigment, such as pyridinium bisretinoid (A2E), are effective generators of the reactive oxygen species, which damage the structure of proteins, lipids and DNA of RPE cells. Increasing with age lipofuscin deposits intensify oxidative stress leading to prooxidative-antioxidative imbalance and irreversible damages in RPE cells. In consequence, their progressive impairment results in death of functionally cell dependent photoreceptors. Therefore, lipofuscin accumulation in the RPE cells is one of the reasons for the development of retina degenerative disorders, such as age-related macular degeneration (AMD), which is the most common cause of irreversible loss of vision over the age of 65.
    No preview · Article · Jan 2011 · Farmaceutyczny Przeglad Naukowy
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    ABSTRACT: The organic composition of the human thigh-bone compact structure was characterized by thermolysis and TMAH- induced thermochemolysis coupled with GC/MS. Tissue sample pyrolysates contained derivatives of benzene, pyridine, pyrrole, phenol, nitrile, sulfur containing compounds, saturated and unsaturated aliphatic hydrocarbons as well as fatty acid derivatives. A large percent content of pyrrole and nitrile derivatives confirmed the quantitative prevalence of protein component in the studied material. Among thermolysis products the individual compound of greatest abudance was pyrrolo-[1,2-α]-piperazine-3,6-dione which it thought to be derived from the collagen dipeptide sequence Pro-Gly, confirming significant content of this protein in bone compact structure. It has also been supported by the findings of the relatively small amounts of sulfur containing products as well as benzene and phenol derivatives, both probably originated from aromatic amino acids which are known to be minor amino acid residues of collagen. Thermochemolysis using TMAH of the studied bone samples enabled to establish fatty acid profile of varying C-chain length (from C12 to C20) with the oleic and palmitic acids as predominating ones. Thermolysis and TMAH termochemolysis coupled with GC/MS are valuable analytical methods to evaluate organic compounds of compact structure of the human bone.
    No preview · Article · Jan 2011 · Farmaceutyczny Przeglad Naukowy
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    Full-text · Article · Nov 2010 · Acta poloniae pharmaceutica
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    ABSTRACT: Matrix metalloproteinases 2 (MMP-2) and 9 (MMP-9) belong to a zinc dependent family of enzymes that degrade components of extracellular matrix. One postulated mechanism by which inositol hexaphosphate (phytic acid, IP6), an ubiquitous plant component, prevents the activation of MMPs may be due to its ability to chelate minerals. The aim of the study was to evaluate the expression profile of MMP-2, MMP-9 and their tissue inhibitors TIMP-1 and TIMP-2 at the mRNA level in human colorectal cancer cell line Caco-2 treated with IP6. A kinetic study of MMP-2, MMP-9 and TIMP-1, TIMP-2 mRNAs was performed after cells treatment with 1; 2.5; 5 mM IP6 for 1, 6, 12 and 24 h. Quantification of genes expression was carried out using real time QRT-PCR technique. The gene encoding MMP-9 was neither constitutively expressed nor induced by IP6 in Caco-2 cells. IP6 at the concentration of 1 mM evoked increase in MMP-2 transcript level, however, its higher doses (2.5; 5 mM) caused a decrease in this gene expression at 1 h incubation. In 24 h lasting culture along with increasing IP6 concentration, the cells expressed lower and lower MMP-2 mRNA level. In response to 1 and 2.5 mM at 6 h, the cells demonstrated an increased transcriptional activity of the TIMP-2 gene which was accompanied by a decrease in TIMP-1 gene transcription. Treatment of cells with 2.5 mM IP6 at 12 h resulted in a strong increase in both TIMP-1 and TIMP-2 expression. The results of this study show that IP6 modulates MMP-2, TIMP-1 and TIMP-2 genes expression in colon cancer cells at the transcriptional level in a way dependent on its concentration and time of interaction.
    Preview · Article · Nov 2010 · Acta poloniae pharmaceutica

  • No preview · Article · Nov 2010 · Acta poloniae pharmaceutica

  • No preview · Article · Nov 2010 · Acta poloniae pharmaceutica
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    ABSTRACT: Familial hypertrophic cardiomyopathy (FHCM) is characterized by an autosomal dominant transmission, left ventricular hypertrophy and myocardial disorganization. So far, 13 genetic loci and more than 130 mutations in ten different genes have been identified. Recent study suggested impaired force production associated with inefficient use of ATP as the main disease mechanism. We performed haplotype analysis with the use of microsatellite markers linked with beta-myosin heavy chain, troponin T, alpha-tropomyosin and cardiac myosin protein C genes in three Polish families with hypertrophic cardiomyopathy (23 individuals). This method is based on the analysis of distribution of the disease in the family and the alleles of chosen microsatellite markers. In two families, the disease was associated with beta-myosin heavy chain gene. We also found a genetic carrier of the mutated gene among children of the patients. In one family the connection of the disease with the mutation in alpha-tropomyosin gene was confirmed, no sudden cardiac deaths were recorded and the degree of myocardial hypertrophy was small.
    No preview · Article · Nov 2010 · Acta poloniae pharmaceutica
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    ABSTRACT: Periodontitis is a destructive disease which is likely to be the result of the activities of different microbial complexes. Recently, sulphate-reducing bacteria (SRB) have been detected in the oral cavity, and they have been found to be common inhabitants of sites showing periodontal destruction. The aim of study was to evaluate the influence of endotoxins of Desulfovibrio desulfuricans bacteria on human gingival fibroblast HGF-1 line. The immunological response of gingival fibroblasts was evaluated by determination of their IL-6 and IL-8 secretion upon treatment with D. desulfuricans intestinal and type strain LPS, sodium butyrate (NaB) and IL-1beta. The amounts of cytokines were estimated by ELISA immunoassay. The influence of LPS and NaB on fibroblast proliferation was determined using the CyQUANT Cell Proliferation Assay Kit. No significant growth inhibition of cells exposed to LPS was observed, except for the culture growing in the presence of intestinal strain endotoxin at the highest concentration (100 microg/ml). The secretion of IL-6 and IL-8 by fibroblasts was increased by D. desulfuricans endotoxins. Cells stimulated with proinflammatory cytokine 1L-1beta showed very high levels of both cytokines secretion. The release of IL-6 and IL-8 by cells in response to LPS and 1L-1beta was modulated by butyric acid. The observed response of gingival fibroblasts to stimulation by endotoxin suggests that D. desulfuricans can be involved in the pathogenesis of periodontitis. Moreover, butyrate present in the oral cavity seems to have immunoregulatory effect on cytokine production by gingival fibroblasts under physiological conditions and during microbe-induced inflammation.
    No preview · Article · Jul 2010 · Archives of oral biology
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    ABSTRACT: Cytomegalovirus (CMV) infection has been suggested to play a role in the development of cardiovascular diseases. It has not yet been established yet whether the possible adverse vascular effect is associated with chronic inflammation process caused by CMV. The aim of our study was to evaluate a possible role of CMV infection in local inflammatory activation in pts with coronary artery disease (CAD). We enrolled 55 patients (mean age 62 years, 42 males, 13 females) with angiographically proven CAD scheduled for CABG surgery. Vessel specimens retrieved from ascending aorta (as a part of routine proximal venous graft development procedure) and peripheral blood mononuclear cells (PBMC) were evaluated for the transcriptional activity of IL-6 and TNF-alpha (the key cytokines involved in atherosclerosis) and for CMV DNA presence. Polymerase chain reaction reaction was performed in order to detect DNA of CMV as well as IL-6 and TNF-alpha transcriptional activity. CMV was present in 67.3% of aortic and in 60% of blood specimens accordingly; median level in aorta tissues: 114.63 +/- 116.54, PBMC: 107.89 +/- 132.39; non statistically significant (NS). An inflammatory response expressed as IL-6 and TNF-alpha transcriptional activity equaled in aorta 159.93 +/- 120.15, 299.55 +/- 154.89 and in PBMC: 190.85 +/- 122.08, 249.64 +/- 32.4; (NS). CMV DNA in PBMC was associated with CMV DNA in aortal tissue p = 0.0049. The analysis revealed positive correlation between IL-6 transcriptional activity and CMV DNA titer in aortic samples R = 0.35, p = 0.036. There were no statistically significant correlations between TNF-alpha transcriptional levels and CMV DNA concentration. Statistical analysis was made by use of Statistica 8.0; StatSoft program. We used arithmetical mean value, standard deviation, Spearmann correlation, X2 and U Mann-Whitney test. A local inflammatory response expressed against CMV could be a marker of longstanding inflammatory response that eventually would cause advanced clinical atherosclerosis. Our findings support the infectious theory and an association between CMV infection and atherosclerosis.
    No preview · Article · Jan 2010 · Wiadomości lekarskie (Warsaw, Poland: 1960)
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    ABSTRACT: Matrix metalloproteinases (MMP) represent the family of endopeptidases that digest components of the extracellular matrix, including basement membranes. Specific tissue inhibitors (TIMP) control metalloproteinases activity. Breakage of matrix barriers enables tumor cells to invade the surrounding tissues and lymph nodes. The aim of the study was to investigate the mRNAs expression of MMP1, MMP2, MMP3, MMP14, and their tissue inhibitors in laryngeal squamous cell carcinoma and adjacent nonneoplastic tissues. Comparison between the mRNA levels of MMPs and TIMPs in tumor samples with lymph node metastasis and without metastatic lymph nodes was made as well. The study included 32 patients with primary laryngeal squamous cell carcinoma. Quantification of genes expression was performed by real time QRT-PCR technique. Expression of all examined MMPs (p<0,01) and TIMPs (p<0,05) was significantly higher in tumor areas. Comparative analysis of all MMP transcripts both in tumor and normal tissues showed the lowest transcriptional activity for MMP1 gene (p<0,01). In tumor and surrounding tissues MMP2, MMP3 and MMP14 genes revealed insignificant difference of transcriptional activity. Expression of TIMP2 mRNA compared with TIMP1 mRNA was higher in tumors (p=0,015), although in adjacent nonneoplastic tissues both mRNAs manifested the same level (p=0,111). Additionally, strong positive correlation between expression of MMP and their tissue inhibitors encoding genes in tumor center (R>0,5; p<0,05) was found. The changes in transcriptional activity of matrix metalloproteinases and their tissue inhibitors may be associated with tumor progression, invasion and metastasis.
    No preview · Article · Jan 2010 · Farmaceutyczny Przeglad Naukowy
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    ABSTRACT: Congenital long QT syndrome (LQTS) is a genetically heterogeneous inherited disease characterized by ECG changes (QT prolongation, abnormal T wave morphology) and predisposition to the occurrence of life-threatening ventricular arrhythmia (torsade de pointes) leading to fainting, cardiac arrest and sudden cardiac death. The most common types of syndrome are due to mutations in the genes KCNQ1 (LQTS1), HERG (LQTS2) and SCN5A (LQTS3) which encode subunits of ion channels in the myocardium. An important role in the diagnosis of LQTS molecular methods (SSCP, DGGE, sequencing) enabling the identification of "silent mutation" carriers with no clinical symptoms, determination of molecular subtype of the disease and confirmation of clinical diagnosis. Screening methods are based on the assumption that the correct and mutant DNA fragments have different electrophoretic mobility in a polyacrylamide gel under non-denaturing conditions (SSCP) or denaturing factor gradient (DGGE). The mSSCP analysis shows higher sensitivity than the classical SSCP due to the use of severalfold temperature changes during the electrophoresis instead of several separations at a different but constant temperatures. The aim of this study was to compare mSSCP and DGGE screening techniques in terms of their ability to detect mutations and polymorphisms in the KCNQ1, HERG, and SCN5A genes. The study involved 35 patients of the Department of Pediatric Cardiology in Katowice-SUM Ligota, whose whole blood was used to isolate genomic DNA as a template for amplification of the fragments of analyzed genes. It was found that of both methods used, DGGE was better to differentiate polymorphisms and mutations in KCNQ1 gene, whereas for genes HERG and SCN5A was preferable one. The results of both analyses are recommended to be confirmed by direct sequence analysis.
    No preview · Article · Jan 2010 · Farmaceutyczny Przeglad Naukowy