Changes in transforming growth factor ß and its receptors' mRNA expression in monocytes from patients with acute coronary syndromes.

INTRODUCTION: Transforming growth factor ß (TGF-ß) is thought to be a vasoprotective cytokine. Numerous reports confirm its significance in blood and plaques. There is, however, a lack of information on the molecular mechanisms involving TGF-ß in circulating inflammatory cells in atherogenesis. sThe aim of the study was to assess gene expression of TGF-ß and its receptors in monocytes from patients with acute coronary syndromes (ACS) and the effect of standard treatment on the studied genes. MATERIAL AND METHODS: The study was carried out in 32 patients with ACS and 15 healthy subjects. Gene expression of TGF-ß and receptors TGF-ßRI and TGF-ßRII was evaluated on day 1 and 5 in the study group and once in controls. The number of mRNA copies isolated from monocytes was assessed by QRT-PCR. RESULTS: Monocytes of ACS patients showed slightly elevated transcriptional activity of TGF-ß1 and its receptors RI and RII genes (0.29 ±0.043 vs. 0.08 ±0.020, p = 0.05; 0.071 ±0.022 vs. 0.036 ±0.023, p < 0.05; 0.134 ±0.020 vs. 0.048 ±0.016, p < 0.05, respectively). After 5-day standard treatment modest reduction of TGF-ßRI expression was observed. The studied genes' expression was unrelated to ejection fraction, myocardial necrosis markers, GRACE score, time from the onset of pain to percutaneous coronary intervention and angiographic findings. Among risk factors family history of CAD was associated with increased TGF-ßRI expression. Moreover, the presence of 4 or more classic risk factors correlated with higher TGF-ßRI expression. CONCLUSIONS: Monocytes of ACS patients demonstrate overexpression of TGF-ß1 and its receptors' genes. Five-day standard treatment downregulated the TGF-ßRI gene but did not affect TGF-ß1 and TGF-ßRII.

[Gene of platelet/endothelial cell adhesion molecule 1 expression profile in patients with acute coronary syndrome and in healthy subjects performed in peripheral blood mononuclear cells using quantitative QRT-PCR reaction]. / Profil ekspresji genu plytkowo-sródblonkowego czynnika adhezyjnego 1 u chorych z rozpoznanym ostrym zespolem wiencowym i w grupie zdrowych oceniany w jednojadrzastych komórkach krwi obwodowej technika QRT-PCR.

UNLABELLED: Cardiovascular diseases including myocardial infarction cause 50 percent of all deaths cases and range before cancer as the cause of death. The term 'ischemic heart disease' covers wide range of diseases including all ischemic conditions of myocardium. It's the most common reason (98%) is atherosclerosis, which causes a restriction or occlusion of a coronary artery lumen resulting in myocardial ischemia. Acute coronary syndromes are caused by the rupture of unstable atherosclerotic plaque resulting in clot formation which blocks the blood vessel. In this process an important role play adhesion molecules triggering adhesion, aggregation and the whole blood coagulation cascade. AIM OF THE STUDY: The aim of the study was to assess PECAM-1 gene expression in peripheral blood mononuclear cells in patients with acute coronary syndrome during first 24 hours of hospitalization in comparison with healthy individuals. MATERIAL AND METHODS: There were 80 subjects included to the study divided into two groups. First group, consecutive patients admitted to the Department of Cardiology due to acute coronary syndrome and the second group, 20 healthy subjects. The total RNA was extracted from peripheral blood mononuclear cell (PBMC) using Chomczynski and Scchaci method. Transcription activity was assessed using commercially available TaqMan Gene Expression Assays. The PECAM-1 gene PCR reaction was preformed with ABI PRISM 7000 Sequence Detector (Applied Biosystems, USA). RESULTS: The comparison of PECAM-1 gene expression revealed statistically significant difference, between increased level of PECAM-1 in patients with acute coronary syndrome and it's lower level in healthy individuals. CONCLUSION: Observed in peripherial blood mononuclear cells increase of PECAM-1 protein gene expression may be responsible for increased adhesion and aggregation process and acute coronary syndrome occurrence.

Gene expression of kinin receptors B1 and B2 in PBMC from patients with cardiac syndrome X.

INTRODUCTION: Cardiac syndrome X (CSX) is defined by typical chest pain, ST segment depression on ECG and normal coronary angiography. Pathology of CSX may involve microvascular dysfunction related to inflammation and abnormal pain sensitivity. Kinins are labile peptides participating in vasodilation, inflammation and pain. Their effects are mediated by two receptors: B1 and B2. The aim of the study was to assess gene expression of kinin receptors in peripheral blood mononuclear cells (PBMC) from patients with CSX. METHODS: The study was carried out in 34 patients with cardiac syndrome X, 13 with unstable angina and ten healthy subjects. Total mRNA was extracted from PBMC and the number of mRNA copies was assessed by quantitive reverse transcriptase polymerase chain reaction. RESULTS AND CONCLUSION: The study showed 7-fold higher transcriptional activity of B1R in CSX vs. control and 3.5 higher vs. UA. B2R expression was 2.5-fold higher in CSX group vs. control and UA, while in the letter two groups it was similar. Such disturbance in kinin signaling may participate in local vasoconstriction and may reflect disturbances in kinin signaling leading to nociceptive disturbances in these patients.

Transcriptional activity of genes encoding interferon gamma (IFNgamma) and its receptor assessed in peripheral blood mononuclear cells in patients with cardiac syndrome X.

BACKGROUND: Cardiac syndrome X is typically characterized by effort induced anginal pain with ST segment depression suggestive of myocardial ischemia and normal coronary arteries at angiography. The possible mechanism that may participate in the pathology of CSX is a microvascular dysfunction related to inflammatory process affecting endothelium. Interferon gamma (IFN-gamma) is an important cytokine in inflammatory reaction. It acts through its specific receptor composed of 2 subunits IFN-gamma R1 (ligand binding) and R2 (signal transduction). The expression and proportion of these subunits influences IFN-gamma activity. The aim of the study was to assess the gene expression of IFN-gamma and its receptors in peripheral blood mononuclear cells (PBMC) from patients with syndrome X. METHODS: The study was carried out in 36 patients aged 44-77 (average 57 years old) with cardiac syndrome X and 23 sex- and age-matched healthy subjects (control group). To evaluate gene expression of IFNgamma and its receptor total mRNA was extracted from peripheral blood mononuclear cells (PBMC) and the number of mRNA copies were assessed by quantitive reverse transcriptase polymerase chain reaction (QRT-PCR). RESULTS: We have not observed statistically significant differences in INFgamma gene expression between studied group and control. Genes encoding IFNgamma receptor subunits showed higher expression in PBMCs from patients with cardiac syndrome X vs control subjects (IFNgammaR1, 97,244 +/- 26,956 c/microg vs 12,120 +/- 2,940 c/microg, p < 0.005, respectively and IFNgammaR2, 129,153 +/- 36,883 c/microg vs 16,445 +/- 2,923 c/microg, p < 0.005, respectively). CONCLUSION: Variation in transcriptional activity of genes encoding INF-gamma receptor subunits may affect function of microvasculature and thereby participate in the pathology of cardiac syndrome X.

[Risk of fetal cytomegaly virus infection depending on number of CMV genome in mother's blood and amniotic fluid]. / Ryzyko zakazenia plodu wirusem cytomegalii w zaleznosci od ilosci kopii genomu CMV w krwi matki i plynie owodniowym.

The cytomegalovirus infection is most common causes of intrauterine infection of the fetus. Using of serologic diagnostic methods the kind of infection is unknown. The aim of the study was assessment of the risk of CMV infection depending of genome account in mother's blood and I amniotic fluid. The study was performed in choosen pregnancies, in which we expected cytomegalovirus infection using serological criteria. In prenatal diagnostic CMV infection using QPCR, the best material is amniotic fluid. Mother's blood assessment of CMV genome count does not make growth diagnostic possibility of the assesement of transmission the infection from mother to fetus.

The transcript expression profile of the leptin receptor-coding gene assayed with the oligonucleotide microarray technique--could this be an anorexia nervosa marker?

Anorexia nervosa is a serious eating disorder with the highest mortality rate of any psychiatric disorder. The DSM-IV classification differentiates two AN types: the restricting type (AN-R) and the binge-eating/purging type (AN-BP). Leptin (LEP) levels can be thought of as a signal to the body of its energy reserves. The leptin receptor (including all its mRNA isoforms) is expressed in many tissues. Our aim was to discover the transcript expression profile of the LEP receptor-coding gene in the peripheral blood mononuclears in AN-R and AN-BP patients. Three young women suffering from Anorexia nervosa (one with AN-BP and two with AN-R) took part in the study, along with three non-anorexic subjects as our reference group. LEP receptor gene expression was examined using the oligonucleotide microarray method (HG-U133A, Affymetrix). The results were normalized using RMAExpress. Next, the accumulation analysis method was used (clustering). Hierarchical clustering resulted in three groups of separate clusters. The first group (cluster I) consisted of AN-R patients. The next group (cluster II) consisted of reference group patients suffering from different psychic disorders not related to eating disorders. Cluster III consisted of two patients--the first with AN-BP and the second with an adaptive disorder.

Transcriptional activity of genes encoding Transforming Growth Factor beta and its receptors in peripheral blood mononuclear cells from patients with acute coronary syndromes.

BACKGROUND: Recent data report altered gene expression of numerous pro- and anti-inflammatory factors involved in pathology of acute coronary syndromes (ACS). Transforming growth factor beta (TGFbeta) signaling is engaged in a wide range of processes. Its effect on vessels seems to be protective due to its anti-inflammatory and anti-atherogenic action. However, it also seems to be engaged in such negative effects as neointima formation and fibrosis. The aim of the study was to assess the expression of the genes encoding TGFbeta and its receptors (type I, II, and III) in patients with ACS. METHODS: The study was carried out on 24 patients with acute coronary syndrome (7 with unstable angina [UA] and 17 with myocardial infarction [MI]) and 10 age-matched healthy subjects (control). To evaluate gene expression of TGFbeta and its receptors total mRNA was extracted from peripheral blood mononuclear cells (PBMC) and the number of mRNA copies were assessed by quantitative reverse transcriptase polymerase chain reaction (QRT-PCR). RESULTS: MI and UA patients demonstrated significantly lower TGFbeta gene expression compared to control (2789+/-418 c/microg vs. 20262+/-2548 c/microg; p<0.001, and 3390+/-518 c/microg vs. 20262+/-2548 c/microg; p<0.001, respectively), as well as noticeably lower transcriptional activity of genes encoding its type I (3295+/-447 c/microg vs. 12859+/-1929 c/microg; p<0.001, and 3258+/-721 c/microg vs. 12859+/-1929 c/microg; p<0.01, respectively) and type II receptors (2364+/-346 c/microg vs. 19003+/-2357 c/microg; p<0.001, and 2680+/-522 c/microg vs. 19003+/-2357 c/microg; p<0.01, respectively). Also, gene expression of the type III receptor was inferior in the studied group compared to the control, although the difference was significant only for the UA group vs. control. Expressions of the studied genes did not differ between patients with MI and those with UA. CONCLUSION: Our report shows that the decreased activity of TGFbeta in patients with ACS is at least partly due altered transcriptional activity of genes encoding both TGFbeta and its receptors, what may be responsible for the evolution of atherosclerotic lesions.

EPR examination of Zn2+ and Cu2+ binding by pigmented soil fungi Cladosporium cladosporioides.

The purpose of this study was to examine the usefulness of electron paramagnetic resonance spectroscopy (EPR) to estimate zinc and copper ions biosorption from the environment by pigmented soil fungi Cladosporium cladosporioides. The existence of a low amount of pheomelanin, besides eumelanin, in C. cladosporioides samples was proved by the analysis of shape of their EPR spectra. Concentration of o-semiquinone free radicals in crude mycelium was 2.4x10(17) spin/g. Changes in free radicals system of C. cladosporioides cultured in the presence of Zn2+ and Cu2+ were analysed. Both magnetic and chemical interactions of zinc and copper ions with free radicals in C. cladosporioides melanin were found. Magnetically interacting diamagnetic Zn2+ ions increased the concentration of o-semiquinone free radicals in melanin existing in C. cladosporioides mycelium, whereas paramagnetic Cu2+ ions decreased this concentration. Chemical interactions of Zn2+ and Cu2+ ions decreased the free radical concentrations in C. cladosporioides melanin. Homogeneously distributed free radicals in C. cladosporioides melanin rise its activity in biosorption processes.

[Diagnostic difficulties of Toxoplasma gondii infection in pregnant women. Is it possible to explain doubts by polymerase chain reaction?]. / Trudnosci w diagnostyce zarazenia ciezarnych Toxoplasma gondii. Czy metoda polimerazowej reakcji lancuchowej moze wyjasnic watpliwosci?

OBJECTIVES: Toxoplasma gondii infection during pregnancy is still a difficult problem in the contemporary perinatology. Difficulties met during interpretation of serological tests carried out in pregnant patients to detect Toxoplasmosis implies more and more frequent use of the Polymerase Chain Reaction (PCR). DESIGN: To evaluate the dependence between serological tests and quantity of the Toxoplasma gondii genomes in mothers' blood and amniotic fluid or neonatal blood, the quantitative PCR (q-PCR) method was applied. MATERIALS AND METHODS: The analysis was performed in 81 pregnant women. Maternal blood, amniotic fluid and newborns' umbilical blood samples were evaluated for the presence of Toxoplasma gondii DNA. IgG and IgM Toxoplasma gondii antibodies were evaluated by the ELISA method. RESULTS: High seroprevalence (51.9%) of the Toxoplasma gondii was confirmed. Toxoplasma gondii genetic material in blood and/or amniotic fluid was found in 33 patients. It was stated that quantity of the protozoa and anti-IgM presence in mothers' blood are the factors influencing significantly the Toxoplasma gondii manifestation in amniotic fluid. CONCLUSION: High suitability of PCR in diagnosis of Toxoplasmosis during pregnancy and vertical transmission was confirmed.

Human cytomegalovirus in patients with systemic lupus erythematosus.

The objective of this study was to determine the frequencies of human cytomegalovirus (HCMV) infection and HCMV genome copy number in blood of consecutive (treated from several months to several years) systemic lupus erythematosus (SLE) patients (22 women). The obtained results were compared to the healthy controls (15 women). All patients fulfilled at least four of the 1982 revised American rheumatism association (ARA) classification criteria for SLE. Our patients demonstrated three or four of the nine possible organ systems involved and most of them had mild SLE with SLE disease activity index (SLEDAI) score < 10 at time when blood samples were collected to detect HCMV. Quantitative analysis of HCMV genome was performed with aid of sequence analyzer ABI PRISM 7,700 Perkin Elmer. Primers and probe were constructed on the basis of IE4 region of HCMV genome. The viral load was expressed as log(10) of calculated HCMV genome copy number. Qualitative analysis revealed that 100% of our SLE patients were infected with HCMV, whereas in the control group only 73% of persons were HCMV positive. Statistically significant difference was demonstrated when the strength of the association between SLE or controls and infection of HCMV was calculated (estimated by Fisher's exact test, P value=0.02). Higher viral DNA copy number was observed in whole blood of SLE patients than in the control group (338.45+/- 221.76 and 229.00+/- 405.61 copies/ml respectively) but did not reach statistical significance level (95% confidence interval from 170.41 to 249.32, P=0.71). Furthermore percentage of patients with HCMV-DNA copy number >2.0 x 10(2) copies/ml was statistically significantly higher than this one in controls. The data show association between HCMV infection and SLE, which should be taken into account during the course of SLE.

Distribution of porcine endogenous retroviruses (PERVs) DNA in organs of a domestic pig.

OBJECTIVES: Domestic pig may serve as the most appropriate organ source for human xenotransplantation in the future. However, there is a serious threat of xenogeneic pathogens transmission, especially porcine endogenous retroviruses (PERVs) which are present in genomes of all pigs. The aim of this study was to monitor the prevalence and distribution of PERV DNA in organs of a domestic pig. METHODS: We used a primer set for a highly conserved fragment of PERV gag sequence to monitor a total PERV DNA copy number and genotype-specific primer sets to study PERV subtypes distribution using Real-Time QPCR (SYBR Green I). RESULTS: Our results showed that PERV DNA was present in all studied pigs, however, most PERV DNA molecules carried numerous mutations thus indicating inability to express functional retroviral particles. The level of PERV DNA in kidney was much higher than in heart (p = 0.007) and in the liver (p = 0.009). CONCLUSIONS: It indicates that kidney is potentially the biggest PERV reservoir which makes it the organ of particular concern in xenotransplantation. We also conclude it is possible to monitor pig herds for individuals with the lowest PERV DNA prevalence, especially lacking PERV-C, and perhaps with only defective PERV proviruses that are unable to express functional RNA.

Effect of apigenin, kaempferol and resveratrol on the expression of interleukin-1beta and tumor necrosis factor-alpha genes in J774.2 macrophages.

Flavonoids have been reported to bring benefits in lowering inflammation, oxidative stress and exert positive effects in cancer and cardiovascular and chronic inflammatory diseases. Apigenin, kaempferol and resveratrol present in fruits, vegetables and grain were investigated for their effect on the synthesis of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) at transcriptional level in lipopolysaccharide (LPS)-stimulated J774.2 macrophages. Apigenin (30 microM), kaempferol (30 microM) and resveratrol (50 microM) significantly decreased the number of TNF-alpha mRNA copies in LPS-activated J774.2 macrophages. Apigenin and kaempferol caused inhibition of IL-1beta gene expression in J774.2 macrophages, but resveratrol was ineffective. These results indicate that apigenin, kaempferol and resveratrol exert inhibitory effects on the TNF-alpha and except for of resveratrol on IL-1beta gene expression in J774.2 macrophages at the transcriptional level. In addition, the studied compounds may be the mediators responsible for protective role of a diet high in fruits and vegetables in the cardiovascular and inflammatory diseases.

Expression of cell survival/death genes: Bcl-2 and Bax at the rate of colon cancer prognosis.

The rate of tumour growth is dependent on the balance between proliferation and apoptosis at all stages of carcinogenesis. Apoptosis inhibition, in turn, depends partly on the balance between expression of two cell death regulatory genes, Bcl-2 and Bax. Colon cancer has long been associated with disturbances in apoptosis regulation. The aim of our study was to determine the expression levels of Bcl-2 and Bax mRNAs in 1 microg sample of total RNA obtained from normal colon and colon adenocarcinoma. This study was intended to evaluate possible differences in Bcl-2 and Bax mRNA levels at particular stages of colon adenocarcinoma classified according to Duke's system. The apoptotic frequency (represented by Bax mRNA copy number) was inversely proportional to the decrease of Bcl-2 gene expression. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was performed to confirm apoptosis.

Familial hypertrophic cardiomyopathy. Insertion-deletion polymorphism of angiotensin-converting enzyme and angiotensin II receptor.

BACKGROUND: Hypertrophic cardiomyopathy (HCM) is a genetic-based disease. Several gene mutations leading to HCM development have been described. AIM: Detailed examination of phenotype and genotype of a family with HCM. METHODS: Clinical and genetic examinations were performed in a family with HCM, in which 3 sick persons with different disease phenotype were found. RESULTS: In all sick persons the same molecular substitution G->A (AGG->AAG) was noticed. It led to substitution Arg780-Lys in exon 21 beta-myosin heavy chain gene, which was responsible for the development of the disease. Insertion- deletion polymorphism analysis in ACE gene revealed D/D (deletion/deletion) genotype in proband and D/I (deletion/ insertion) phenotype in his mother and sister, who were heterozygous. Polymorphism A1166C analysis in AT1 gene revealed the presence of genotype A/A in proband and A/C in his mother and sister. In proband and his sister a very similar phenotype was observed, whereas they had different polymorphism for ACE gene and angiotensin 1 receptor gene. In sick proband's mother, who had phenotype different to her children, the same polymorphism as in his daughter was noticed. CONCLUSIONS: In the described family with HCM, different phenotype and polymorphism of ACE and AT1 genes were found.

Immunolocalization and expression of kinin B1R and B2R receptors in human inflammatory bowel disease.

Bradykinin is a mediator of inflammation, responsible for pain, vasodilation, and capillary permeability. Bradykinin receptor 1 (B(1)R) and bradykinin receptor 2 (B(2)R) are G protein-coupled receptors that mediate kinin effects. The latter is constitutive and rapidly desensitized; the former is induced by inflammatory cytokines and resistant to densensitization. The distribution of bradykinin receptors in human intestinal tissue was studied in patients with inflammatory bowel disease (IBD), namely ulcerative colitis (UC) and Crohn's disease (CD). Both B(2)R and B(1)R proteins are expressed in the epithelial cells of normal and IBD intestines. B(1)R protein is visualized in macrophages at the center of granulomas in CD. B(2)R protein is normally present in the apexes of enterocytes in the basal area and intracellularly in inflammatory tissue. In contrast, B(1)R protein is found in the basal area of enterocytes in normal intestine but in the apical portion of enterocytes in inflamed tissue. B(1)R protein is significantly increased in both active UC and CD intestines compared with controls. In patients with active UC, B(1)R mRNA is significantly higher than B(2)R mRNA. However, in inactive UC patients, the B(1)R and B(2)R mRNA did not differ significantly. Thus bradykinin receptors in IBD may reflect intestinal inflammation. Increased B(1)R gene and protein expression in active IBD provides a structural basis of the important role of bradykinin in chronic inflammation.

Effect of kaempferol on the production and gene expression of monocyte chemoattractant protein-1 in J774.2 macrophages.

Monocyte chemoattractant protein-1 (MCP-1) is produced by activated macrophages, and is involved in pathogenesis of cardiovascular and neurodegenerative disorders. There is a need to develop drugs that inhibit excessive infiltration of monocytes and lymphocytes to the arterial wall and central nervous system. The aim of this study was to evaluate the effect of kaempferol on the (MCP-1) gene expression and MCP-1 protein release by J774.2 macrophage cultures in vitro. Kaempferol given both before and after lipopolysaccharide (LPS) administration reduced secretion of MCP-1. Kaempferol administered before LPS stimulation significantly decreased the number of copies of MCP-1 mRNA. The results suggest that kaempferol inhibits MCP-1 production at the transcriptional level, and that this is an additional anti-inflammatory mechanism of action of this flavonoid.

Butyrate-induced differentiation of colon cancer cells is PKC and JNK dependent.

Butyric acid, a short-chain fatty acid physiologically present in human large gut, is derived from bacterial fermentation of complex carbohydrates. It has been shown to reduce the growth and motility of colon cancer cell lines and to induce cell differentiation and apoptosis. Apoptosis is considered a result of normal colonocyte terminal differentiation in vivo. The aim of this study was to characterize the cellular mechanisms regulating differentiation of colon cancer cells stimulated with sodium butyrate (NaB). The two human colon cancer cell lines Caco-2 and HT-29 were treated with NaB at physiologically relevant concentrations. Alkaline phosphatase (ALP) activity, a marker of colonocyte differentiation, was increased 48 hr after treatment with 1 mM NaB. Higher doses of NaB (5 and 10 mM) induced apoptosis of the cells and failed to stimulate the colonocyte differentiation. Therefore, we assumed that butyrate augments cell differentiation and induces apoptosis, acting via various intracellular mechanisms, and butyrate-mediated programmed cell death cannot be considered a consequence of colonocyte terminal differentiation. The effect of NaB on ALP activity was significantly attenuated in the presence of inhibitors of protein kinase C and JNK. Inhibition of MEK-ERK signal transduction pathways augmented the impact of butyrate on colonocyte differentiation. These results suggest that butyrate could influence the colonocyte differentiation via modulation of the activity of cellular protein kinases and signal transduction.

[Quantitative analysis of sigma genes expression in Mycobacterium tuberculosis cultures exposed to rifampicin and isoniazid]. / Analiza profilu ekspresji genów kodujacych czynniki sigma w hodowlach Mycobacterium tuberculosis eksponowanych na ryfampicyne oraz izoniazyd.

The aim of the study was quantitative analysis of five genes encoding Mycobacterium tuberculosis sigma factors sigA, sigE, sigF, sigH, and sigI as well as the 85B reference gene known as the mycobacterial viability marker, in cultures exposed to rifampicin and isoniazid. The mRN levels were assessed using QRT-PCR technique, in the automated system of real time quantification with the ABI PRISM 7700 Sequence Detector System (TaqMan). The number of each analyzed gene transcript copies was expressed as a number of mRNA per 1 eg of isolated total RNA. In cultures exposed to the tested chemicals the number of 85B mRNA copies declined as compared to the controls (without tested chemicals). There was no detectable expression of sigA and sigI in the control cultures. Both, rifampicin and isoniazid induced expression of sigA and sigI genes. The sigE gene expression increased during exposure to isoniazid and decreased under rifampicin exposure conditions. The sigF mRNA was detected neither in the control culture, nor in cultures exposed to rifampicin or isoniazid. Both tested chemicals caused decrease of sigH expression.

[Beta2-agonists--what does their chemical structure determine?]. / Beta2-agonisci--o czym decyduje ich struktura chemiczna?

The paper presents the structure and functional relationship of beta2-agonists and the beta2-adrenoceptor. The human beta2-adrenoceptor is a member of the 7-transmembrane family of receptors. Most of the actions of the beta2-receptor are mediated through the Gs protein and the cAMP-dependent PKA system. Alternative cAMP-independent pathways affected following beta2-receptor activation have also been described. Beta2-agonists have been used as bronchodilator agents in the treatment of asthma since the development of inhaled isoprenaline preparations in 1961. Over the years, these agents have been markedly improved through the development of short-acting beta2 selective agents followed by long-acting beta2 selective agents with prolonged duration of action. Efforts directed towards improving the pharmacodynamic and pharmacokinetic properties, including the long-acting and selective beta2-adrenoceptor agonists, included two major pathways of modifying the basic structure of the catecholamines, which are described in this paper. The pharmacological activity usually resides in the (R)-enantiomer. The kinetics of beta2-agonist-stimulated bronchodilation in asthma is determined by the differences in the molecular mechanism of beta2-agonists action. Regulation of the beta2-receptor is influenced by its desensitization following exposure to high concentrations of or repeated challenges with agonists, and up-regulation which can be induced by glucocorticoids and thyroid hormones.

Sapheno-femoral junction pathology: molecular mechanism of saphenous vein incompetence.

A molecular mechanism responsible for varicose vein occurrence was investigated. The role of potential cell cycle regulator p21 and programmed cell death in the pathology leading to the proximal long saphenous vein (LSV) incompetence was investigated. Proximal LSV specimens were obtained from 40 patients with primary varicose veins who had undergone crossectomy. The expression of the p21, p53, and fas encoding genes was investigated by the means of real-time RT-QPCR. Immunostaining for gene product presence, proliferating cell nuclear antigen (PCNA), and apoptotic cells (TUNEL assay) was carried out. The results were compared to the control healthy vein specimens and correlated with pathologic examination findings (of the valve and vein structure). A significant increase in p21, p53, and fas mRNA expression were reported in the proximal incompetent veins. The expression of p21 correlated with expression of p53 (r = 0.658; p<0.05) and negative correlation between media apoptotic index and p21 mRNA expression was found (r = -0.493; p<0.05). Decrease in the muscular component within the media and disturbances of the local structure in the incompetent LSVs were reported. Fas overexpression did not correlate with p53 expression level and did not correlate with apoptotic cell number in the respective vein layers. PCNA-positive cells were present more frequently in the media of the control veins, especially in young subjects. Apoptosis downregulation, cell cycle inhibition and smooth muscle cell hypertrophy are important factors influencing vein wall disturbances related to sapheno-femoral junction incompetence.