Heterozygote Subjects Research Articles

The pathophysiology of Wilson's disease visualized: Ahuman 64 Cu PET study.

Background and AimsWilson’s disease (WD) is a genetic disease with systemic accumulation of copper that leads to symptoms from the liver and brain. However, the underlying defects in copper transport kinetics are only partly understood. We sought to quantify hepatic copper turnover in patients with WD compared with heterozygote and control subjects using PET with copper‐64 (64Cu) as a tracer. Furthermore, we assessed the diagnostic potential of the method.Approach and ResultsNine patients with WD, 5 healthy heterozygote subjects, and 8 healthy controls were injected with an i.v. bolus of 64Cu followed by a 90‐min dynamic PET scan of the liver and static whole‐body PET/CT scans after 1.5, 6, and 20 h. Blood 64Cu concentrations were measured in parallel. Hepatic copper retention and redistribution were evaluated by standardized uptake values (SUVs). At 90 min, hepatic SUVs were similar in the three groups. In contrast, at 20 h postinjection, the SUV in WD patients (mean ± SEM, 31 ± 4) was higher than in heterozygotes (24 ± 3) and controls (21 ± 4; p < 0.001). An SUV‐ratio of hepatic 64Cu concentration at 20 and 1.5 h completely discriminated between WD patients and control groups (p < 0.0001; ANOVA). By Patlak analysis of the initial 90 min of the PET scan, the steady‐state hepatic clearance of 64Cu was estimated to be slightly lower in patients with WD than in controls (p = 0.04).Conclusions 64Cu PET imaging enables visualization and quantification of the hepatic copper retention characteristic for WD patients. This method represents a valuable tool for future studies of WD pathophysiology, and may assist the development of therapies, and accurate diagnosis.

Computational Inference, Validation, and Analysis of 5'UTR-Leader Sequences of Alleles of Immunoglobulin Heavy Chain Variable Genes.

Upstream and downstream sequences of immunoglobulin genes may affect the expression of such genes. However, these sequences are rarely studied or characterized in most studies of immunoglobulin repertoires. Inference from large, rearranged immunoglobulin transcriptome data sets offers an opportunity to define the upstream regions (5’-untranslated regions and leader sequences). We have now established a new data pre-processing procedure to eliminate artifacts caused by a 5’-RACE library generation process, reanalyzed a previously studied data set defining human immunoglobulin heavy chain genes, and identified novel upstream regions, as well as previously identified upstream regions that may have been identified in error. Upstream sequences were also identified for a set of previously uncharacterized germline gene alleles. Several novel upstream region variants were validated, for instance by their segregation to a single haplotype in heterozygotic subjects. SNPs representing several sequence variants were identified from population data. Finally, based on the outcomes of the analysis, we define a set of testable hypotheses with respect to the placement of particular alleles in complex IGHV locus haplotypes, and discuss the evolutionary relatedness of particular heavy chain variable genes based on sequences of their upstream regions.

Laron syndrome - A historical perspective.

Laron Syndrome (LS) [OMIm#262500], or primary GH insensitivity, was first described in 1966 in consanguineous Jewish families from Yemen. LS is characterized by a typical phenotype that includes dwarfism, obesity and hypogenitalism. The disease is caused by deletions or mutations of the GH-receptor gene, causing high serum GH and low IGF-I serum levels. We studied 75 patients from childhood to adult age. After early hypoglycemia due to the progressive obesity, patients tend to develop glucose intolerance and diabetes. The treatment is by recombinant IGF-I, which improves the height and restores some of the metabolic parameters. An unexpected finding was that patients homozygous for GH-R defects are protected from malignancy lifelong, not so heterozygotes or double heterozygote subjects. We estimate that there are at least 500 patients worldwide, unfortunately only few treated.

G-protein coupled-receptor 65 5´UTR gene polymorphism in the pathogenesis of systemic lupus erythematosus

Systemic lupus erythematosus (SLE) is a complex autoimmune disease with unknown etiology. G-protein-coupled receptor 65 (GPR65) candidates as an SLE-locus for functioning in T cell receptor-mediated self-reactive T cell death in the thymus. This is also involved in anti-inflammatory actions and apoptosis as remarkable features of autoimmune diseases. This study investigated the relationship between the rs10139328 polymorphism at the 5´UTR of a GPR65 gene and SLE. This case-control study consisted of 102 SLE patients (98 females, 4 males) and 118 age- and gender-matched healthy controls (113 females, 5 males). Genotyping of the rs10139328 polymorphism was determined using an amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Data was analyzed using SPSS software. The Pearson chi-square was the test of choice for assessing the association between the rs10139328 polymorphism and SLE. The probable influences of sunlight and family history on SLE were evaluated by performing logistic regression. Except for one heterozygote subject among the control group, the study population was homozygote for the selected polymorphism. No statistical difference was seen in genotype distribution between the cases and the controls (P> 0.05). Statistical analysis revealed that sun exposure directly increased SLE susceptibility (P < 0.001). Having a family history of SLE increased the risk of disease occurrence by more than two times (OR = 2.38, 95% CI: 1.28 - 4.41, P= 0.006). The results of the current study do not support the importance of the studied polymorphism in a GPR65 gene in the pathogenesis of SLE among southwestern Iranian patients.

Relationships between cytokine (IL-6 and TGF-β1) gene polymorphisms and chromosomal damage in hospital workers

Cytokine gene polymorphisms have been found to be associated with a pre-disposition to a variety of diseases, including inflammatory and cancer diseases. The present study evaluated the influence of six cytokine gene polymorphisms on the level of genomic damage observed in peripheral blood lymphocytes from hospital pathologists chronically exposed to low doses of different xenobiotics. Lymphocytes from 50 pathologists and 50 control subjects were recruited and analyzed in Sister Chromatid Exchange (SCE) and Chromosomal Aberrations (CA) assays. The frequencies of six cytokine gene polymorphisms and their relationships with the cytogenetic damage levels were also evaluated. The results indicated that significant differences were found between pathologists and controls in terms of SCE frequency (p < 0.001) and RI values (p < 0.001), as well as in terms of CA and cells with aberrations (p < 0.001). No associations were found between all analyzed cytokine gene polymorphisms and CA frequency in both pathologists and control groups. Vice versa, among pathologists, homozygote individuals for the IL-6 G allele showed a significantly (p = 0.017) lower frequency of SCE with respect to heterozygote subjects. Similarly, for TGFβ1 codon 10 locus, homozygote for T allele and heterozygote TC subjects showed a significantly (p = 0.021) lower frequency of SCE with respect to homozygote CC individuals. Among controls, no significant differences were found in the frequency of SCE between genotypes at all loci. Based on these results, we speculate that high circulating levels of a pro-inflammatory cytokine like IL-6 and lower levels of the immunosuppressant cytokine TGFβ1 could be associated directly with a longer duration and/or greater intensity of inflammatory processes, and indirectly with significantly higher levels of genomic damage.

A family with various symptomatology suggestive of Anderson–Fabry disease and a genetic polymorphism of alpha galactosidase A gene

BackgroundAnderson/Fabry disease expresses a wide range of clinical variability in patients that it is possible to explain referring to a genetic variability with numerous mutations described in the literature (more than 600). MethodsWe report some clinical cases of some members of a Sicilian family to express phenotypical variability of this disease in subjects with the same genetic mutation ResultsThe first case was a 59-year-old female. Brain MRI revealed right frontal periventricular white matter of likely vascular-degenerative origin. The proband's alpha galactosidase A activity was 3.7nmol/mL/h. Molecular genetics revealed a polymorphism: −10 C>T; IVS 2-76_80del5; IVS4-16 A>G; IVS6-22 C>T. The second case was a 30year-old male affected by acroparesthesias and hypoidrosis since he was an adolescent. Renal impairment was first detected at age 29; it began with high plasma levels of creatinine and microalbuminuria date. The third case was a 41year-old daughter that presented with acroparesthesias, hypoidrosis since she was very young. The patient's alpha galactosidase A activity was 4.1nmol/mL/h, in whole blood, which is compatible with heterozygote subject for Fabry's disease or healthy control. The fourth case was a male grandson of the proband, 9year-old child. He had a classic gastrointestinal involvement. He complained of recurrent abdominal pain, post prandial bloating and pain. This child's enzyme activity was 1.65nmol/mL/h. In cases 2, 3, and 4, molecular genetics revealed a polymorphism: −10 C>T; IVS 2-76_80del5; IVS4-16 A>G; IVS6-22 C<T DiscussionA recent study reported that IVS4+68 A>G, IVS6-22C>T polymorphisms occurred in 8.9% and 3.7% of the subjects respectively, and the significance of this haplotype in FD pathology remains unknown but possibly suggestive of Anderson/Fabry disease.

I19 Homozygosity in HD Patients - Case Reports

Background The homozygosity for CAG expansion is a very rare finding in HD patients. It is assumed, that homozygous cases show similar age at onset in comparison to the heterozygotes with the same expansion length but the disease progression seems to be more rapid with pronounced psychiatric symptoms, probably due to wider neuronal degeneration. We present the clinical course of two HD patients harbouring two mutant alleles, diagnosed in the Institute of Psychiatry and Neurology (IPN), Warsaw, Poland. Case history Two patients were clinically and genetically evaluated while being enrolled into the REGISTRY EHDN project. They showed different clinical presentation and course of the disease. The first subject (28 y.o.) with 38 and 63 CAG repeats presented with juvenile HD phenotype. Symptoms began at age of 12, with rapid development of dystonia and rigidity (Baseline TMS 74), the presence of epileptic seizures, cognitive impairment, severe irritability and obsessive-compulsive disorder. The second patient (44 y.o.) with 40 and 42 CAG repeats showed first symptoms at the age of 40. Until now the patient is affected with a mild motor symptoms (TMS 23). The depression with suicidal ideations and psychotic symptoms (delusions) seem to be the most important clinical problem. The rate of decline of cognitive abilities is not different from these observed in heterozygous patients. Conclusions Our study on two homozygous HD cases revealed, that the clinical course of motor symptoms in them does not differ significantly from the relevant cohorts of heterozygotic subjects. However, it confirmed suggestions from other reports that the psychiatric component of HD clinical presentation may be more severe in homozygous subjects.

283. HIV Protected Autologous Zinc Finger Nuclease Driven CCR5 Modified CD4 T-Cells CCR5 (SB-728-T) Reduce HIV Viral Load in CCR5 Δ32 Heterozygote Subjects During Treatment Interruption (TI): Correlates of Effect, and Effect of Cytoxan Pre-Conditioning Regimen

283. HIV Protected Autologous Zinc Finger Nuclease Driven CCR5 Modified CD4 T-Cells CCR5 (SB-728-T) Reduce HIV Viral Load in CCR5 Δ32 Heterozygote Subjects During Treatment Interruption (TI): Correlates of Effect, and Effect of Cytoxan Pre-Conditioning Regimen

Association of gene variants in TLR4 and IL-6 genes with Perthes disease

Perthes disease is idiopathic avascular osteonecrosis of the hip in children, with unknown etiology. Inflammation is present during development of Perthes disease and it is known that this process influences bone remodeling. Since genetic studies related to inflammation have not been performed in Perthes disease so far, the aim of this study was to analyze the association of frequencies of genetic variants of immune response genes, toll-like receptor 4 (TLR4) and interleukin-6 (IL-6), with this disease. The study cohort consisted of 37 patients with Perthes disease and 50 healthy controls. Polymorphisms of well described inflammatory mediators: TLR4 (Asp299Gly, Thr39911e) and 11-6 (G-174C, G-597A) were determined by polymerase chain reaction restriction fragment length polymorphism method. Results IL-6 G-174C and G-597A polymorphisms were in complete linkage disequilibrium. A statistically significant increase of heterozygote subjects for IL-6 G-174C/G-597A was found in controls in comparison to Perthes patient group (p = 0.047, OR = 2.49, 95% CI = 1.00-6.21). Also, the patient group for IL-6 G-174C/G-597A polymorphisms was not in Hardy-Weinberg equilibrium. No statistically significant differences were found between patient and control groups for TLR4 analyzed polymorphisms. A stratified analysis by the age at disease onset also did not reveal any significant difference for all analyzed polymorphisms. Conclusion Our study revealed that heterozygote subjects for the IL-6 G-174C/G-597A polymorphisms were significantly overrepresented in the control group than in the Perthes patient group. Consequently, we concluded that children who are heterozygous for these polymorphisms have a lower chance of developing Perthes disease than carriers of both homozygote genotypes.

A Novel Tetranucleotide Repeat Polymorphism Within KCNQ1OT1 Confers Risk for Hepatocellular Carcinoma

KCNQ1 overlapping transcript 1 (KCNQ1OT1), a long noncoding RNA responsible for silencing a cluster of genes in cis, has been shown to be involved in multiple cancers. However, much remains unclear of how KCNQ1OT1 contributes to carcinogenesis. By thoroughly analyzing 510 hepatocellular carcinoma (HCC) cases and 1014 healthy controls in a Chinese population, we identified a novel short tandem repeat (STR) polymorphism (rs35622507) within the KCNQ1OT1 coding region and evaluated its association with HCC susceptibility. Logistic regression analysis showed that compared with individuals carrying the homozygote 10-10 genotype, those heterozygote subjects who carry only one allele 10 had a significantly decreased risk of HCC (adjusted odds ratio [OR]=0.67, 95% confidence interval [CI]=0.53-0.86, p=0.0009), with the risk decreased even further in those without allele 10 (adjusted OR=0.38, 95% CI=0.21-0.69, p=0.0005). Furthermore, genotype-phenotype correlation studies using four hepatoma cell lines support a significant association between STR genotypes and the expression of KCNQ1OT1. Cell lines without allele 10 conferred a 20.9-33.3-fold higher expression of KCNQ1OT1. Meanwhile, KCNQ1OT1 expression was reversely correlated with the expression of the cyclin-dependent kinase inhibitor 1C (CDKN1C), a tumor suppressor gene located within the CDKN1C/KCNQ1OT1 imprinted region, in three hepatoma cell lines. Finally, in silico prediction suggested that different alleles could alter the local structure of KCNQ1OT1. Taken together, our findings suggest that the STR polymorphism within KCNQ1OT1 contributes to hepatocarcinogenesis, possibly by affecting KCNQ1OT1 and CDKN1C expression through a structure-dependent mechanism. The replication of our studies and further functional studies are needed to validate our hypothesis and understand the roles of KCNQ1OT1 polymorphisms in predisposition for HCC.

Hypocretin (orexin) neuropeptide precursor gene, HCRT, polymorphisms in early-onset narcolepsy with cataplexy

To test if the hypocretin (orexin) neuropeptide precursor (HCRT) gene, HCRT, mutations are implicated in the development of narcolepsy with cataplexy deficiency in young children. The entire HCRT gene and ~2000 bp promoter region was first sequenced in 181 patients and 153 controls, and rare polymorphisms including three nonsynonymous amino acid changes were identified. Next the 557 bp region of exon 2 harboring the three nonsynonymous changes was sequenced in an additional 298 early-onset subjects and in 148 control samples. A previously known common polymorphism (rs760282) and nine rare novel polymorphisms were identified in subjects and controls without significant differences. Two nonsynonymous exon 2 substitutions (+977 H54A, +979 G55R) were detected in two subjects with early onset at 7 and 6 years, respectively, but were not found in any controls. These substitutions are not likely to vastly change peptide binding to hypocretin receptors. One additional exon 2 substitution (+1019, K68R) was found in two patients and one control. Additional sequencing that focused on exon 2 showed additional subjects and controls with the +1019 K68R polymorphism and without significant differences between the subjects and the control. Segregation of two of these three nonsynonymous single nucleotide polymorphisms (SNPs) were observed from unaffected parents to offspring. Sequencing of a large number of early-onset narcolepsy subjects revealed three novel nonsynonymous substitutions within the preprohypocretin protein, two of which were only found in patients with early-onset narcolepsy but are not likely to be functionally significant, especially in heterozygote subjects.

Abstract 167: A comparison of 25-OH-vitamin D levels and VDR polymorphisms in healthy and patients with breast intraepithelial neoplasia or cancer .

Abstract Background: Vitamin D is mainly synthesized in the skin through UVB irradiation, while a minor intake comes from the diet. Vitamin D is converted in the liver to 25-hydroxyvitamin D-(25(OH)D) and circulating levels of this metabolite is an appropriate index of vitamin D status. A further hydroxylation occurs to produce the biologically active metabolite 1,25-dihydroxyvitamin D. Epidemiological studies have suggested a role for vitamin D in cancer prevention showing an inverse relationship between incidence of cancers and mortality and 25(OH)D plasma levels. Vitamin D exerts its activity through a nuclear receptor (VDR) that regulates the expression of multiple genes, including genes for cell cycle regulation, differentiation, and apoptosis Methods: We have collected plasma from pre and post menopausal healthy women (HW) and newly diagnosed patients with an intraepithelial neoplasia (IEN) or breast carcinoma (BC). We measured circulating levels of 25(OH)D3 using LC-MS/MS. The median values were then correlated with the disease status. In a subgroup, we performed a correlation with VDR polymorphisms (SNPs) and disease recurrences. BsmI, FokI, TaqI and ApaI SNPs were determined using the Applied Biosystems’ Taqman Allelic Discrimination Assay. Results: 25(OH)D3 was measured in 249 HW, 175 subjects with IEN and 168 BC patients undergoing surgery. Age and BMI were taken into account for adjustments the median age being 51, 46 and 59 years and median BMI being 24, 23 and 25.6 kg/m2 in the HW, IEN and BC categories, respectively. Median 25(OH)D3 were respectively: 37.2, (inter-quartile range, IQR 25.0-49.7) 43 (IQR 30.0-60.0) and 42 (IQR 30.0-60.0) nmol/L respectively. The LS means with BC + IEN combined versus HW were 43 (IQR 40.8-45.2) vs 46.4 (IQR 43.9-49.0) nmol/L. The levels of 25(OH)D3 are statistical different between healthy versus affected subjects (p = 0.05). We also conducted an exploratory analysis in a subgroup of 228 subjects to evaluate the correlation of VDR polymorphisms and the disease free survival (DFS for breast recurrence or new event). With a follow-up of 160 months and a total of 70 breast events, the homozygote and heterozygote subjects for VDR ApaI polymorphism had a worst DFS compared to the wild type ones (log rank test p= 0.04).Conclusions: our data confirm the possible role of the vitamin D and its receptor in the carcinogenesis process. It is not clear when 25(OH)D level starts to decline and thus may influence the disease progression. In accordance with other research groups the ApaI polymorphism may have, together with other VDR SNPs, an influence on BC incidence and recurrence. Further correlations and analysis of the VDR genotype are ongoing. A prospective study to clarify is recommended the issue of the timing of 25(OH)D levels measurement is warranted. Citation Format: Davide Serrano, Harriet Johansson, Debora Macis, Ernst Lien, Jennifer Gjerde, Antonella Puccio, Sara Gandini, Aliana Guerrieri-Gonzaga, Irene Feroce, Matteo Lazzeroni, Andrea Decensi, Bernardo Bonanni. A comparison of 25-OH-vitamin D levels and VDR polymorphisms in healthy and patients with breast intraepithelial neoplasia or cancer . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 167. doi:10.1158/1538-7445.AM2013-167

Low protein Z levels but not the protein Z gene G79A polymorphism are a risk factor for ischemic stroke

Background Protein Z (PZ) is a vitamin K-dependent plasma protein that plays a role in both pro-and anticoagulant pathways, but its exact physiological function remains unclear. The aim of this study was to determine the association between the G79A PZ gene polymorphism in intron F, PZ levels and the occurrence of ischemic stroke. Methods We performed a case–control study in 118 Caucasian patients with first ever ischemic stroke or TIA confirmed by CT, and 113 age-and sex-matched population controls. Venous blood samples for PZ levels were collected 7 to 14 days and 3 months after stroke onset. Estimates of relative risk (odds ratios) were adjusted for vascular risk factors. Results The adjusted relative risk of ischemic stroke associated with PZ levels in the lowest quartile versus the highest quartile was 3.0 (95% CI: 1.1–8.7) at 7–14 days, and 5.1 (95% CI: 1.2–21.9) at 3 months after the stroke. PZ levels in the convalescent sample were significantly lower than in the acute sample. In the convalescent sample, odds ratios increased with lower quartiles of protein Z level (test for trend p = 0.02). Thirty-nine patients (33%) and 32 (28%) controls were heterozygous for the G79A PZ gene polymorphism and 4 (3%) patients and 4 (4%) controls had the AA-genotype. The PZ levels were significantly lower in subjects with the AA-genotype and intermediate in heterozygote subjects. The odds ratio of ischemic stroke associated with A-allele carriers versus GG-homozygotes was 1.2 (95% CI: 0.7–2.1). Conclusion No association between the G79A PZ gene polymorphism and the occurrence of stroke was observed. However, low PZ levels are independently associated with an increased risk of ischemic stroke.

Peroxisome proliferator-activated receptor α polymorphisms and postprandial lipemia in healthy men

Peroxisome proliferator-activated receptor alpha (PPARalpha) is a ligand-dependent transcription factor that plays a key role in lipid and glucose homeostasis. This study evaluated whether variants of PPARalpha are associated with postprandial lipemia. Subjects were given a single fat load composed of 60% calories as fat, 15% as protein, and 25% as carbohydrate. Blood was drawn every hour from baseline to 6 h, then every 2.5 h to 11 h to determine triglyceride (TG) levels. The minor allele of the nonsynonymous p.Leu162Val variant was associated with higher fasting total cholesterol, LDL-cholesterol, and apolipoprotein B. There were no significant associations with all of the postprandial parameters examined. Conversely, the noncoding variant c.140+5435T>C was not associated with fasting lipid concentrations but was significantly associated with decreased postprandial TG and cholesterol in the small TG-rich lipoprotein particle. Although the minor allele carriers displayed lower mean concentrations of TG and cholesterol throughout the postprandial period, the differences were most pronounced in the latter period. These data suggest that PPARalpha variants may modulate the risk of cardiovascular disease by influencing both fasting and postprandial lipid concentrations.

High frequency of vitamin B12 deficiency in asymptomatic individuals homozygous to MTHFR C677T mutation is associated with endothelial dysfunction and homocysteinemia

The aim of this study was to examine the association of homozygosity for the methylenetetrahydrofolate reductase (MTHFR) C677T mutation and vitamin B12 deficiency in 360 asymptomatic individuals and to investigate forearm endothelial function in C677T homozygotes. MTHFR C677T mutation and levels of vitamin B12, folic acid, and homocysteine were measured in study participants. Frequency of homozygosity for the C677T mutation was 67/360 (18.6%). Homocysteine levels were elevated in homozygous compared with heterozygous subjects or those without the mutation (20.6 +/- 18.8 vs. 9.4 +/- 3.2 mumol/l; P < 0.0001). The number of subjects with vitamin B12 deficiency (<150 pmol/l) was significantly higher among the homozygote than the heterozygote subjects or subjects without mutation [20/67 (29.8%) vs. 27/293 (9.2%); P < 0.0001]. Homozygote subjects had 4.2 times higher probability of having B12 deficiency (95% confidence interval = 2.1-8.3). Forearm endothelial function was assessed in 33 homozygote and 12 control subjects. Abnormal endothelial function was observed in homozygous subjects and was worse in homozygote subjects with vitamin B12 deficiency. Endothelial function was normalized after B12 and folic acid treatment. We found that homozygosity for the C677T mutation is strongly associated with B12 deficiency. Coexistence of homozygosity for the C677T mutation and B12 deficiency is associated with endothelial dysfunction and can be corrected with vitamin B12 and folic acid treatment.

Hyperhomocysteinemia and Heat Shock Proteins. Epidemiological Study in a General Population.

Records and objectives: This study is planned with the objective of identifying new efficacious risk vascular markers. For this work the hyperhomocysteinemia as independent thrombotic vascular risk factor, atherosclerosis as inflammatory illness and Heat Shock Proteins (HSP70i) as molecules likely involved in the vascular damage are considered.Subjects and methods: Workers of the HGGM have volunteered and given their consent. 53 male (M) and 54 female (F) were studied randomly. Anamnesis and epidemiological questionnaire were performed. Arterial pressure was measured and blood samples were taken for neutrophils leukocytes isolation by gradient, biochemical study and DNA extraction. We performed quantification of homocysteine (tHcy)by HPLC; Reactive-C-Protein (RCP), serum and intraleukocitary HSP70i, and HSP70i antibodies by ELISA; molecular study of the polymorphism of C677T of the enzyme metilentetrahidropholatereductase (MTHFR) by PCR-RFLP. Task force of coronary risk was applied for the population classification.Results: The studied population present [tHcy] ≥ 11 umol/L in the 12% of the cases. The mutation C677T appears in heterozygote subjects (CT) in the 45% of the cases and in homozygote subjects in the 17%.Subjects were classified into 3 groups: G-0, without vascular risk factors, n=77 (32M and 45F); G-1, with a moderated risk factor (>10%) but without illness n=26 (18M and 8F) and G-2, with an evident atherosclerotic illness n=4 (3M and 1F)Conclusions: The following correlations have been proved:Inverse (p<0.05) between [tHcy] and [HSP70i] in neutrophils in females G-0 and males G1 wild type (CC)Inverse (p<0.05) between [tHcy] and serum [HSP70i] in males G-1 homozygotes (TT)Direct (p<0.05) between [tHcy] and [RCP] in females G-0 heterozygotes (CT)Direct (p<0.05) between [tHcy] and cholesterol in females G-0 homozygotes (TT)Direct (p<0.01) between [tHcy] and HTA in males G-0 heterozygotes (CT)This study has been partially financed by FIS 03/1308

The dependence of serum interleukin-6 level on PPAR-alpha polymorphism in men with coronary atherosclerosis

Background The association between L162V polymorphism in the gene for peroxisome proliferator activated receptor (PPAR) alpha and the development of coronary heart disease was examined. Methods PPAR-alpha polymorphism was determined in 48 men with angiographically confirmed coronary atherosclerosis and in 51 healthy men. Results The frequency of the V allele of the L162V polymorphism was four times higher in men with atherosclerosis (0.25 in studied group and 0.06 in controls). The polymorphism was not associated with changes in body mass index, lipid pattern, serum adhesion molecules, or vasoactive agents concentrations. The effect of the polymorphism on the serum interleukin-6 level (IL-6) was observed ( p < 0.01). The serum IL-6 level was higher in homozygotic than in heterozygotic subjects ( p < 0.02). Multivariate regression analysis showed the existence of a relationship between simvastatin therapy and serum IL-6 level ( r = 0.83; p < 0.05) in the homozygotic men. While in homozygotic patients with atherosclerosis a negative linear correlation between serum IL-6 and NO concentration was shown, in heterozygotic men positive correlations between IL-6 or HDL cholesterol and adhesion molecule levels were found. Conclusion L162V polymorphism in the gene for PPAR-alpha seems to be associated with atherosclerosis through a mechanism including regulation of the IL-6 level.

Evidence for Selective Expression of the p53 Codon 72 Polymorphs: Implications in Cancer Development

Polymorphism at codon 72 of p53 results in either the arginine or proline form of p53, whose functional significance in carcinogenesis is controversial. We have investigated if the expression of these p53 polymorphs is selectively regulated, using mRNA from peripheral blood of healthy Asian (Chinese) and the Caucasian (Polish) arginine/proline (arg/pro) heterozygote subjects. Asians were found to preferentially express the pro allele whereas the Caucasians preferentially express the arg allele. On the contrary, about 75% of the heterozygote Chinese breast cancer patients preferentially expressed the arg allele, which rarely contained any somatic mutations. Moreover, histologically normal tissues from Chinese heterozygote breast cancer patients showed selective expression of the arg allele, in contrast to the preferential expression of the pro allele in heterozygote healthy normal breast tissues. Together, the data suggest that the expression of the different p53 polymorphs is selectively regulated in different ethnic populations, and that the arg allele is activated during cancer development in Asians. Thus, the expression status of the p53 polymorphs, rather than the genotypic status, might be a useful indicator for cancer susceptibility.

Effects of combined CYP2C9 and 2C19 polymorphism on the pharmacokinetics of glibenclamide in Chinese subjects

Background Recent in vitro studies showed that both CYP2C9 and 2C19 contributed to the metabolism of glibenclamide (glyburide). This study investigates the relative in vivo contribution of CYP2C9 and 2C19 to glibenclamide pharmacokinetics. Methods Eighteen healthy male Chinese subjects were divided into 3 groups: CYP2C19 EM/CYP2C9*1/*1 (group 1, n=6), CYP2C19 PM/CYP2C9*1/*1 (group 2, n=6), and CYP2C19 EM/CYP2C9*1/*3 (group 3, n=6). Subject received a single oral dose of 5 mg glibenclamide, and multiple blood samples were collected over 12h. Results No significant differences in glibenclamide pharmacokinetics were observed between CYP2C19 EM and PM subjects (group 1 vs 2). However, the plasma glibenclamide concentration was significantly higher in CYP2C9*1/*3 than CYP2C9*1/*1 subjects regardless of CYP2C19 genotype (group 3 vs 1 or 2), and their respective AUC0-∞ were 1.03±0.50, 0.46±0.13 and 0.57±0.11 μg•h/ml (ANOVA, p<0.05), and respective t1/2 were 3.58± 1.25, 2.09± 0.22 and 2.24± 0.27h (ANOVA p<0.05). In addition, 50% of CYP2C9*1/*3 subjects and 17% of CYP2C9*1/*1 subjects developed blood glucose levels below 3 mmol/L and required oral glucose administration. Conclusion CYP2C9 but not 2C19 polymorphism appears to exert dominant influence on glibenclamide metabolism. In CYP2C9*1/*3 heterozygote subjects, the pharmacokinetics and pharmacodynamics of glibenclamide were markedly changed, and the glibenclamide dosage may need to be adjusted to reduce the risk of hypoglycemia. Clinical Pharmacology & Therapeutics (2005) 77, P23–P23; doi: 10.1016/j.clpt.2004.11.087

Molecular cytogenetic parameters in fibroblasts from patients and carriers of xeroderma pigmentosum

Xeroderma pigmentosum (XP) is a rare autosomal recessive syndrome. Laboratory investigations have failed to detect any consistent anomaly in cells from XP heterozygotic subjects, although examples of behavior intermediate between normal and XP cells have been reported. To estimate random aneuploidy we applied fluorescence in situ hybridization (FISH) with α-satellite probes for chromosomes 8 and 9 and replication pattern for TP53 ( p53), ERBB2 ( HER-2/neu), and MYCN ( N-MYC) loci and for the imprinted SNRPN locus. A significantly higher rate of aneuploidy rate was observed in XP patients and carriers than in controls. The asynchrony pattern was significantly higher in XP carriers and patients with all three coding loci analyzed and significantly lower in XP patients and carriers with the imprinted locus SNRPN than in the control group. Molecular cytogenetic parameters such as random aneuploidy and replication pattern, which are known to reflect chromosomal instability, may be part of the tumorigenesis process. In XP patients and carriers, this genetic instability may represent a potential for developing malignancies.