Search

Background: Chronic inflammation of the arterial wall plays a crucial role in the pathogenesis of atherosclerosis. Cyclooxygenase-2 (COX-2) is a key enzyme in the synthesis of proinflammatory prostanoids. At least two of the common genetic polymorphisms of the COX-2 gene have phenotypic effects: G-765C (rs20417) and T8473C (rs5275)., Aim: To assess the relation of G-765C and T8473C COX-2 polymorphisms to clinical and angiographic characteristics of patients with coronary artery disease (CAD)., Material and Methods: The study comprised 186 consecutive patients with angiographically defined CAD (> or =70% stenosis of > or =1 coronary artery). The study population were divided into two groups: A-123 patients with stable angina (mean age, 62.6 +/- 11.2 years; 30.1% women), and B-63 patients with unstable angina (mean age, 64.0 +/- 10.8 years; 19.0% women). The controls comprised 70 individuals without symptoms of CAD (mean age, 37.6 +/- 9.9 years; 57.1% women). Results: No significant differences were observed in -765C and 8473C allele frequencies between the patients with CAD and control subjects. In CAD patients, the studied COX-2 polymorphisms were not significantly associated with the age of the onset of symptoms and clinical presentation of CAD. In the B group, a difference was observed within the frequency of significant (>50%) left main coronary artery stenosis (LMCAS) and/or three-vessel CAD (3-CAD) between the -765C allele carriers and 765G 765G homozygotes (14.3% vs. 49.0%; p=0.044). In the CAD patients (group A and group B) the prevalence of LMCAS and/or 3-CAD was significantly lower among 365C allele carriers (22.8% vs. 40.3%:, Conclusions: There were no significant differences in -765C and 8473C allele frequencies between patients with CAD and subjects without symptoms of CAD; In patients with CAD, COX-2 G-765C and T8473C polymorphisms had no significant association with the age of the onset of symptoms and clinical presentation of ischaemic heart disease; The G-765C COX-2 polymorphism is associated with less frequent occurrence of multivessel CAD in the studied population. p=0.021

In recent years, the analysis of X-linked short tandem repeats (X-STR), beside autosomal and Y-chromosomal STR loci, has become widely used in forensic genetic investigations. The usefulness of X-STRs markers in forensic medicine is confirmed by their high biostatistical parameters obtained for different populations. Such population studies performed on particular ethnic groups allow for demonstrating the presence of, rare genetic variants that are not included in the allelic ladders of commercially available multiplex kits. The presence of these alleles can increase the power of evidence. On the other hand, the off-ladder alleles can be sometimes difficult to interpret. In this paper, X-STRs analysis was performed in a population sample of 200 unrelated females and males from the Southern Poland using Investigator Argus X-12 Kit. Seven rare off-ladder alleles were encountered: DXS10074*15.2, DXS10079*24, DXS10146*38.2, DXS10146*47.2, DXS10148*17, DXS10148*21.1 and DXS10148*22 that were not previously reported in the Polish population. Additionally, genetic transmission of these genetic variants was ascertained.

Introduction: Inflammatory mediators, including prostanoids produced by inducible cyclooxygenase-2 (COX-2), play a significant role in the development of atherosclerosis. A regulatory region of COX-2 gene has a common -765G>C polymorphism. Functional effects of this polymorphism and its association with atherosclerosis phenotypes have not been fully understood., Objectives: The aim of the study was to evaluate the association between COX-2 -765G>C polymorphism and the inflammatory response in patients with stable CAD., Patients and Methods: We studied systemic prostaglandin E2 (PGE2) metabolism, the levels of soluble CD163 (sCD163) in serum (a marker of monocyte/macrophage activation), and COX-2 -765G>C polymorphism in patients with stable CAD. We also tested the patients for functional effects of COX-2 -765G>C polymorphism using cell lines, using the constructs in which red fluorescent protein expression was controlled by a large segment of COX-2 regulatory region., Results: Patients with stable CAD carrying the variant allele -765C allele had increased urinary excretion of PGE2 metabolite and higher serum levels of sCD163 than patients carrying the -765G allele. In contrast to these clinical findings, in vitro functional studies demonstrated that the -765C variant allele was less responsive than -765G allele to a wide range of COX-2 inducers., Conclusions: A substantial part of total PGE2 biosynthesis is contributed by activated monocytes/macrophages in stable CAD. The exact mechanism of activation of this pathway in CAD requires further research because of the conflicting results on COX-2 -765G>C polymorphism provided by clinical studies and in vitro functional studies.

Low-affinity IgE receptor gene (FCER2) rs3760687 polymorphism was found to be associated with differential binding affinity of transcription factors Sp1 and Sp3 leading to altered transcriptional activity. Haplotypic interaction of functional FCER2 polymorphisms (rs28364072, rs2228137 and rs3760687) might potentially provide a background for genotype-phenotype associations previously observed for some rather non-functional FCER2 variants.

During a routine paternity casework performed with an automated genotyping using the AmpFISTR Identifiler kit, a lack of paternal allele segregation in D21S11 and off-ladder allele in D19S433 locus was observed. This raised suspicion of mutation because the other systems showed transmission of putative father's alleles to the child. To achieve the recommended value of paternity index (PI), the range of analysis was extended by additional autosomal loci (Penta D and Penta E) and haplotype of the chromosome Y. No further exclusions were observed. Additionally, a new PowerPlex ESI 17 kit was used. By comparison, the results common for both the multiplex kits confirmed the presence of the mutation in D21S11 locus. In locus D19S433, a rare variant D19S433*7 allele was evidenced in the putative father and the child. This variant allele was included in size range of allelic ladder in the PowerPlex ESI 17 kit, but not in AmpFfSTR Identifiler kit. The variant D19S433*7 allele was not reported in the Polish population before. ThlSTRe PowerPlex ESI 17 genotyping kit has two advantages, i.e. introduction of 6 loci not included in the Identifiler kit, and extended ranges of allelic ladders. It seems that a careful scrutiny of fluorescent signals following electrophoretic separation is essential to detect rare variant alleles and to avoid misinterpretation of the results.

To seek for novel rare and/or sporadic mutations within genomic neighborhood of common -344 C>T (rs2427827) FCER1A proximal promoter polymorphism, which by its prevalence could have masked the presence of less frequent genetic variants in our previous single-stranded conformational polymorphism (SSCP) mutational search study, SSCP screening was repeated using primers fixing -344 C>T variant. The genomic region surrounding -344 C>T polymorphism was confirmed to be fairly conservative and only a single sporadic mutation (present in 1 out of 196 chromosomes), a 6-bp deletion -262 to 257 del CTAGAC, was found. From the methodological point of view, we demonstrated a successful detection of a short-length polymorphism using SSCP screening in a population, in which the same genomic region contained frequent single-nucleotide polymorphic variant. In conjunction with subsequent cloning of aberrantly migrating PCR products and SSCP-driven indirect sequencing of the clones, this method offers a superior (to direct sequencing of PCR products) detection of rare mutations. The cost-effective method applied by us enables a reliable characterization of infrequent (thus present only in heterozygotic form) short-length variance of the sequence which are difficult to interpret by direct sequencing.

Our knowledge on the variability of FCER1A gene encoding for alpha-subunit of the high-affinity immunoglobulin E receptor (FcepsilonRI) that plays a central role in the pathogenesis of allergy and related disorders, has been recently much extended. Last findings from FCER1A mutational screening and genetic association studies, followed by functional analyses of the polymorphisms, are briefly summarized in this mini-review. The association between FCER1A gene variants and total serum IgE levels seems especially interesting and, supported by functional analyses of polymorphisms, may provide a rationale for pharmacogenetic studies on anti-IgE therapy that indirectly suppresses FcepsilonRI expression.

The arachidonic acid metabolites participate in development of coronary artery disease (CAD) and the plaque's instability. We assessed two common genetic polymorphisms: of cyclooxygenase-2 (COX-2) (COX2.8473, rs5275) and prostaglandin EP2 receptor gene (uS5, rs708494) in patients with CAD. Out of 1,368 patients screened by coronary arteriography, two groups fulfilled the entry criteria and were studied: stable coronary disease (sCAD, n = 125) and acute coronary syndromes (ACS, n = 63). They did not differ in the main characteristics. All patients were on aspirin at least seven days prior to the study. In 70 control subjects, the same genotypes were ascertained, expression of cyclooxygenases in peripheral blood monocytes was assessed by flow cytometry, and in-vitro biosynthesis of PGE(2) was measured by mass spectrometry. COX-2 CC homozygotes (variant allele), were more common, while EP2 GG homozygotes (wild-type) were less common in ACS (p = 0.03 and p = 0.017) than in the sCAD group. A combined genotype characterized by the presence of the wild-type COX2.8743T allele and the wild type homozygous EP2uS5 genotype (TT or CT | GG) decreased risk ratio of ACS in CAD patients (relative risk 0.41; 95% confidence interval 0.21-0.81). COX-2 polymorphism in control subjects did not affect the enzyme expression or PGE(2) production by peripheral blood monocytes, but production of PGE(2) increased by 40.1% in the subjects homozygous for EP2 receptor allele uS5A following lipopolysaccharide stimulation. In conclusion, the combined COX-2 (COX2.8473) and the EP2 receptor (uS5) genotypes seem to influence CAD stability, but in peripheral blood monocytes only EP2 receptor modulates PGE(2) production.

Two FCER1A gene proximal promoter polymorphisms, -344C > T and -95T > C, both associated with total serum IgE and/or allergic disorders in Caucasians and East Asians, were shown to influence the gene expression in additive manner. In face of the rarity of other proximal promoter variants in Caucasians or their lack in East Asians, future investigations of the FCER1A locus in these ethnic groups should probably focus on three common haplotypes of the common -344C > T and -95T > C polymorphisms.

Objective: The diallelic glycoprotein IIIa polymorphism P1A1/A2 was attributed to be an inherited risk factor for coronary events. Whether this polymorphism affects response to aspirin in patients with coronary artery disease is not known., Methods: We assessed thrombin generation (prothrombin fragment F1+2) in consecutive blood samples collected from bleeding-time wounds in 28 men with coronary artery disease; P1A2 carriers, n=9; P1A1/A1, n=19. Thrombin generation and bleeding time were measured before and after 2 weeks of aspirin 300 mg/day., Results: Aspirin-depressed thrombin generation in A1 homozygotes (p=0.04), but not in A2 carriers. Bleeding time after aspirin was also prolonged in A1 subjects only (p=0.02)., Conclusion: Genotyping for glycoprotein IIIa polymorphism might be helpful in predicting antithrombotic action of aspirin in secondary prevention of coronary artery disease.

Background: The high-affinity receptor for immunoglobulin-E (IgE) (FcepsilonRI) plays a major role in the pathogenesis of allergy, but there are only two published studies on its alpha subunit (FcepsilonRIalpha) genetic variability in allergic diseases., Aims of the Study: Mutational screening in the region of the FcepsilonRIalpha gene promoter and the first exon with subsequent genetic variability assessment in allergic patients and a random population sample., Methods: Allergic subjects were individuals with asthma or urticaria. Age- and sex-matched controls were randomly selected from a large population sample. Mutational screening was performed using a single-stranded conformational polymorphism and subsequent sequencing. Detected polymorphisms were genotyped by restriction fragment length polymorphism. Total serum IgE was measured in allergic subjects and controls. Skin prick tests, blood eosinophil count and aspirin challenge test were performed only in the subjects. A subgroup of the subjects was further characterized by autologous serum skin test, histamine release test, Phadiatop and IgE antibodies against staphylococcal enterotoxins., Results: Two linked polymorphisms -344 C>T and -95 T>C were found within the FcepsilonRIalpha gene. The allele -344 T frequency was 0.45 vs 0.37 (P = 0.33), and the allele -95 C frequency was 0.26 in subjects vs 0.30 in controls (P = 0.62). Serum IgE was significantly higher in subjects homozygous for the -344T allele (TT genotype) than in those carrying the -344 C allele (CT or CC genotype; P = 0.003), but this association was not detectable in controls., Conclusions: Our findings of genotype-related differences in IgE levels in allergic patients suggest an impact of -344 C>T but not -95 T>C gene polymorphism of FcepsilonRIalpha on total levels of IgE. The genetic variability in FcepsilonRIalpha at the -344 nucleotide of its regulatory sequence, though not related to atopy, predicts higher levels of the immunoglobulin.

Etiology of chronic obstructive pulmonary disease remains unknown but, despite some inconsistencies in reports on inflammatory cells, mediators and proteases involved in the pathogenesis of chronic obstructive pulmonary disease, genetic risk factors were proposed as a cause of susceptibility to the disease. Results of many studies suggested polygenic inheritance, with the genetic component consisting of several genes of a small effect each, rather than of single major gene. We are going to review the clinical importance of alpha-1 antitrypsin, glutathione S-transferase, microsomal epoxide hydrolase, matrix metalloproteinase, tumor necrosis factor-a, alpha-1 antichymotrypsin, alpha 2-macroglobulin, cytochrome P4501A1, heme oxygenase-1 genes polymorphisms associated with susceptibility and progression of the chronic obstructive pulmonary disease.

The proinflammatory cytokines, like interleukine-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha), have been suggested to play a role in the development of atherosclerotic conditions, e.g. peripheral arterial occlusive disease (PAOD). Several genetic polymorphisms were described within the cluster of the IL-1 genes. The allelic variant C+3953T of the interleukin-1beta (IL-1beta) gene correlated with the cytokine level in vitro. The aim of this study was to estimate frequency of IL-1beta +3953 polymorphism in the Polish population and to test for its association with PAOD. We genotyped IL-1beta gene in 115 PAOD patients and 153 healthy controls. Genotypes distribution and allelic frequencies of C+3953T polymorphism did not differ between PAOD and controls. Thus, no association was observed between functional IL-1beta polymorphism and PAOD in our study. Frequency of the variant +3953T allele in Polish subjects (0.258) was similar to this found in other populations.

Background: Prostaglandins, generated via the COX pathways, are essential mediators of inflammation in bronchial asthma. The promoter polymorphism of COX-2 gene (G-765C), which might affect binding of transcription factors, has recently been described, Objective: To study distribution and function of the genetic COX-2 variant in patients with asthma compared with healthy controls., Methods: Three groups of adults were studied: (1) patients with aspirin-induced asthma (AIA; n=112), (2) asthmatic patients who tolerated aspirin (ATA; n=198), and (3) a random population sample from city of Krakow (n=547). The COX-2 promoter region was genotyped for the G-765C polymorphism. Ex vivo production of prostaglandin E2 and prostaglandin D2 by peripheral blood monocytes was measured., Results: In the 2 asthmatic groups, the G-765C allele frequency was similar (AIA, 0.18; ATA, 0.19) and did not differ from that of controls (0.17). In asthmatic women, but not in men, CC homozygotes were overrepresented compared with controls (odds ratio, 3.08; 95% CI, 1.35-6.63; P=.01). There was no relationship between genotype and FEV1, serum IgE, blood eosinophil count, or duration of the disease. In AIA but not in ATA patients, CC homozygosity was associated with more severe course of the disease, as reflected by need for oral corticotherapy. Production of 2 prostaglandins by monocytes was more than 10-fold higher in CC than in GG homozygotes, and the magnitude of this difference was not changed by LPS stimulation., Conclusion: In asthma, the COX-2 -765C homozygosity is associated with female sex. The CC homozygosity has functional effects resulting in increased capacity of monocytes to produce prostaglandins.

We analyzed the role of the C677T polymorphism of the 5,10-methylenetetrahydrofolate and the A66G polymorphism of the methionine synthase reductase genes as risk factors for occurrence of spina bifida. The studied population included 106 mothers and 104 children from affected families, and a control group of 100 adults. We found statistically significant differences between the occurrence of the homozygosity in these polymorphisms in the groups of mothers and children with thoracolumbal defects (C677T polymorphism) and lumbosacral defects (A66G polymorphism). We postulate that these polymorphisms should be regarded as independent risk factors for spina bifida.

Frequently an inherited predisposition to thrombosis remains clinically silent until an additional environmental factor intervenes. The present study aimed to assess distribution of inherited risk factors of venous thrombosis in patients with venous thromboembolism (VTE). The prevalences of factor V Leiden (FV Leiden), prothrombin factor II G20210A (FII G20210A), C677T and A1298C of methylenetetrahydrofolate reductase (MTHFR) mutations were studied in 149 VTE patients and 100 controls. The following key risks were established: previous deep venous thrombosis or pulmonary embolism (23.5%), bed rest (34.2%), immobilisation of lower limb (10.1%), hospitalisation (30.9%) and obesity (28.9%). In 29 (19%) patients and in three (3%) controls FV Leiden was found. A significant association between VTE and FV Leiden was established. There were six (4%) carriers of the FII G20210A among VTE patients and one in the controls. No associations between VTE and MTHFR polymorphisms (C677T, A1298C) were found. In three of 149 patients both FV Leiden and FII G20210A polymorphisms were observed. The mean protein C activity was slightly, though nonsignificantly, smaller in VTE patients. In conclusion, there was a positive association between venous thromboembolism and factor V Leiden. Only a weak trend favouring a relationship between prothrombin factor II G20210A and venous thrombolism was present. No associations between common polymorphisms of methylenetetrahydrofolate reductase and venous thromboembolism were found.

We report on a rare allelic variant of the TH01 STR locus. Within this commonly used tetranucleotide repeat system, we identified and characterized by denaturating gel electrophoresis an allele of the size corresponding to 10.3 repeats, analogous to the most common 9.3 allele. The novel allele segregated from a putative father to the child in a paternity case. We discuss the advantage of high resolution second generation multiplex systems (SGM) using internal size standards during electrophoresis.