HTRA1) gene rs11200638 G→A polymorphism and LOC387715/ ARMS2 gene rs10490924 G→T polymorphisms, and to evaluate the magnitude of the gene effect and the possible genetic mode of action.To examine the association of age-related macular degeneration (AMD) with HtrA serine peptidase 1 (HTRA1 (rs11200638) and LOC387715/ ARMS2 (rs10490924) gene polymorphisms and AMD. We performed a meta-analysis conducted with Stata software, version 9.0.We searched the US National Library of Medicine’s PubMed, Embase, OMIM, ISI Web of Science, and CNKI databases in a systematic manner to retrieve all genetic association studies on the HTRA1 gene rs11200638 G→A polymorphism had 2.243 and 8.669 times the risk of developing AMD, respectively, when compared with those who carry the GG genotype. Individuals carrying the TT and TG genotypes of LOC387715/ ARMS2 gene rs10490924 G→T polymorphism had 7.512 and 2.353 times the risk of developing AMD, respectively, compared with those who carry GG genotype. These results suggested a “moderate” codominant, multiplicative genetic mode; that is, both HTRA1 rs11200638 G→A polymorphism and LOC387715/ARMS2 rs10490924 G→T polymorphism play important roles in the pathogenesis of AMD. We found no evidence of publication bias. Between-study heterogeneity was found in both allele-based analysis and genotype-based analysis.Individuals who carried the AA and AG genotypes of HTRA1 rs11200638 G→A polymorphism and LOC387715/ARMS2 rs10490924 G→T polymorphism play important roles in AMD. Gene-gene and gene-environmental interactions, as well as precise mechanisms underlying common variants in the HTRA1 gene and LOC387715/ ARMS2 gene, potentially increase the risk of AMD and need further exploration. National Advisory Council).Age-related macular degeneration (AMD) is a neurodegenerative disease that leads to visual impairment and accounts for half of all cases of registered blindness in Western individuals older than 65 years of age -14. TherCFH) gene on chromosome 1q31 has been revealed as the first major AMD-susceptibility gene, perhaps accounting for about 30%–50% of AMD patients. The CFH Y402H variant in the CFH gene has also been identified as a causal polymorphism in studies of populations other than those of European and North American origin [CFH Y402H polymorphism and AMD [CFH may independently contribute to AMD susceptibility [LOC387715/age-related maculopathy susceptibility 2 [ARMS2]), and high-temperature requirement factor A1 (HTRA1/PRSS11) [AMD is a clinically heterogeneous and genetically complex disease, with multiple environmental and genetic risk factors involved -25. Whiln origin -78, and n origin  has conf and AMD ,81, sugg and AMD -84 of CFtibility ,76,82-84/PRSS11) ,76,82-84HTRA1 gene spans a 53,366-base region on chromosome 10q26 ; it encodes a member of a family of serine proteases expressed in both mouse and human retinas [rs11200638) in the promoter region of the HTRA1 gene was found to be significantly associated with susceptibility to AMD in studies of Caucasian populations in the US [LOC387715/ARMS2, has recently been identified. LOC387715/ARMS2 encodes a deduced 107–amino acid protein with nine predicted phosphorylation sites and a molecular mass of 12 kDa. Real-time (RT)-PCR analysis demonstrated that LOC387715/ARMS2 transcripts were expressed in the retina and in a variety of other tissues and cell lines. Transfection experiments in mammalian cells localized the protein to the mitochondrial outer membrane [rs10490924) in exon 1 of the LOC387715/ARMS2 gene was the most likely susceptibility allele of AMD. Since an individual study may not have sufficient statistical robustness to confirm the association between HTRA1 and LOC387715/ARMS2 gene polymorphisms and AMD, we considered that a meta-analysis that combined data from all published studies would provide a more accurate estimate of the extent of association, leading to less risk of false-positive results [HTRA1 rs11200638 G→A polymorphism, the LOC387715/ARMS2 rs10490924 G→T polymorphism, and AMD. We attempted to estimate the strength of the genetic association with AMD, as well as the genetic mode of action, and to gauge the extent of heterogeneity in the strength of the associations among different studies.The  retinas ,86, and  retinas . HTRA1 a retinas . Overexp retinas . During n the US -97, Centn the US , France n the US , and then the US ; of Eastn the US -104 and n the US -107; andn the US . Anothermembrane . Up to nmembrane  concludePubMed, Embase, OMIM, ISI Web of Science, and Chinese National Knowledge Infrastructure (CNKI) databases in a systematic manner to retrieve all genetic association studies on the HTRA1 (rs11200638) and LOC387715/ARMS2 (rs10490924) polymorphisms and AMD published before April 2008. The search strategy was based on a combination of the terms (HtrA serine peptidase 1 or HTRA1), (age-related maculopathy susceptibility 2 or LOC387715), and (age-related macular degeneration or AMD). The references of all computer-identified publications were searched for additional studies, and the PubMed option ‘‘Related Articles’’ was also used to search for potentially relevant papers. Searches were performed by two independent reviewers (B.Z. and J.Y.). We included all published articles regardless the language of publication.We searched the US National Library of Medicine’s HTRA1 polymorphism (rs11200638), respectively, were A and G and AA, AG, and GG. 3) The alleles and genotypes for the LOC387715/ARMS2 polymorphism (rs10490924), respectively, were G and T and GG, GT, and TT. 4) The numbers of subjects possessing each allele and genotype in the AMD and control groups were available. 5) In the case of multiple publications from the same study group, the most complete and recent results were used. We set no restriction on the source of controls . For those studies where AMD was graded , the gradings were combined into a single AMD group.Studies were included if they met the following criteria: 1) The study reported original data from case-control or cohort studies. 2) The alleles and genotypes for the Data were extracted independently by two investigators (B.Z. and J.Y.), who used recommended guidelines to report on meta-analyses of observational studies . The folHTRA1 and LOC387715/ARMS2 polymorphisms and AMD, and to avoid excessive comparisons, we calculated the odds ratio by two methods: allele comparison , and comparing the risk-variant homozygotes and heterozygotes with wild homozygotes . We estimated and characterized the prevalence of the risk allele with only the data from controls. When we analyzed genotype data in the meta-analysis, zero cell counts were assigned a fixed value . In addition, we calculated the population attributable risk (PAR) of the risk allele according to the Chang et al. [We used the odds ratio as the metric of choice and this was estimated for each study. To explore the possible association between We first compared the alleles for cases and controls to detect overall differences and genetic association. Allele frequencies were computed for studies reporting only genotypic data. Pooled odds ratios were computed two times: by the fixed effects model of Mantel and Haenszel , and by HTRA1 rs11200638 G→A polymorphism, the LOC387715/ARMS2 rs10490924 G-to -T polymorphism, and AMD was based on the comparisons between risk-variant homozygotes and heterozygotes versus wild homozygotes so that the strength of the genetic association and the genetic mode of action could be identified exactly. Once an overall gene effect was confirmed, the genotype effects and genetic model were estimated by using the genetic model-free approach suggested by Minelli et al. [OR1 and OR2. The logarithm (log) odds ratios were modeled on the basis of both between- and within-study variations. A stochastic parameter lambda (λ), equal to the ratio of log OR2 and log OR1, was also computed [Our primary genetic analysis of the i et al. , in whici et al.  was usedcomputed . The parF) suggested by Trikalinos et al. [F value for each study. Predicted genotype frequencies were estimated and then used to replace the observed frequencies in the summary analysis of magnitude and the genetic model.We examined the deviations from the HWE in control populations for each study by using the exact method . For alls et al. . In brieQ statistic [I2 statistic, which describes the percentage of variability in point estimates due to sample heterogeneity rather than sampling error [I2 values larger than 75% were considered to represent a “notable” heterogeneity [In sensitivity analysis, we estimated between-study heterogeneity across all eligible comparisons using Cochran’s tatistic . We alsong error ,120, andng error . I2 valuogeneity . Publicaogeneity  and cumuogeneity . In the meta, metan, metabias, metacum, and metareg commands, except the Bayesian method of genotype-based analysis. We fitted the Bayesian models by using Markov chain Monte Carlo methods with a Bayesian framework and performed our inferences using WinBUGS 1.4.3  [CrI) describes a range in which it is probable that an unknown quantity lies within this interval. A “burn-in” of 10,000 iterations is performed for models, followed by 50,000 iterations for parameter estimates. A p value less than 0.05 was considered statistically significant.All analyses were conducted with Stata software, version 9.0  , using ton 2003) , taking HTRA1 rs11200638 G→A polymorphism and AMD [A total of 29 studies were identified based on our search strategies, of which 13 studies -106,108  and AMD ,130-132, and AMD ,91 were  and AMD , and one and AMD .HTRA1 rs11200638 G→A polymorphism and AMD are presented in LOC387715/ARMS2 rs10490924 G→T polymorphism and AMD are presented in Detailed characteristics of the 13 included studies on the association between HTRA1 rs11200638 G→A polymorphism among controls was 32.33% , and was significantly higher in Asians than in Caucasians . The frequency of the risk allele T in the LOC387715/ARMS2 rs10490924 G→T polymorphism among controls was 25.17% , and was also significantly higher in Asians than in Caucasians .Data from the control groups were used to calculate the summary allele frequency. The frequency of the risk allele A in the HTRA1 rs11200638 G→A polymorphism and AMD [OR=2.910, 95% CI: 2.552, 3.318; Q=25.769, p=0.012, I2=53.4%). The magnitude of the effect was similar for Asians  and Caucasians . However, there was significantly greater between-study heterogeneity among Caucasians  than Asians . Excluding and adjusting two studies [OR=3.043, 95% CI: 2.725, 3.397; Q=14.318, p=0.216, I2=23.2%). We did not find any evidence of publication bias in the eligible studies . All of the 13 studies were included to evaluate the association between the  and AMD -106,108. studies ,97 with LOC387715/ ARMS2 rs10490924 G→T polymorphism and AMD was also evaluated. As shown in OR=2.734, 95% CI: 2.366, 3.158; Q=80.195, p=0.000, I2=78.8%). The magnitude of the effect was similar between Asians  and Caucasians . There was also a significant difference between-study heterogeneity among Caucasians  as opposed to Asians . The association between the HTRA1 rs11200638 G→A polymorphism between case and control groups is presented in OR1) and AG versus GG (OR2) were calculated for each study. The genotype frequency of the LOC387715/ARMS2 rs10490924 G→T polymorphism between the case and control groups is presented in OR1) and TG versus GG (OR2) were calculated for each study.The genotype frequency of the OR1=8.469, 95% CrI: 6.766, 10.710) and heterozygous variant AG genotype  of the HTRA1 rs11200638 G→A polymorphism. A moderate level of between-study heterogeneity  was found for the homozygous AA genotype and no between-study heterogeneity  was found for the heterozygous AG genotype. The estimated parameter λ was 0.378 , which suggested a moderate codominant genetic mode of action. When we removed the two studies [OR1, OR2, and λ of 9.257 , 2.334 , and 0.380 , respectively; however, no significant between-study heterogeneity was found for either the homozygous AA genotype  or the heterozygous AG genotype . The pooled estimates also remained similar after adjusting HW disequilibrium by coefficient F .In our primary analysis, multivariate meta-analysis was conducted to estimate the pooled risk and there was a significantly increased risk of AMD among individuals with both homozygous variant AA genotype  and heterozygous variant TG genotype  of the LOC387715/ ARMS2 rs10490924 G→T polymorphism. The estimated parameter for λ was 0.426 , which suggested a moderate codominant genetic mode of action.Multivariate meta-analysis also showed that there was a significantly increased risk of AMD among individuals with both the homozygous variant TT genotype  and Caucasian populations . A moderate degree of between-study heterogeneity was found for the AA homozygous genotype among both Asians  and Caucasians , but no significant between-study heterogeneity was found for the AG homozygous genotype among either population . For the LOC387715/ ARMS2 rs10490924 G→T polymorphism, a moderate level of between-study heterogeneity was found for the TT homozygous genotype among Caucasians  and for the TG heterozygous genotype among both Asians  and Caucasians ; however, no significant degree of between-study heterogeneity was found for the TT homozygous genotype among Asians .For the OR2 . We performed stratification analysis on wet AMD and the combined AMD of the HTRA1 rs11200638 G→A polymorphism, and found a considerable difference in effects between wet AMD  and combined AMD . This stratification exhibited no between-study heterogeneity for either OR1  or OR2  for combined AMD, and found a moderate degree of between-study heterogeneity for OR1  and non-significant between-study heterogeneity for OR2  of the wet AMD  was significantly associated with log  wet AMD .LOC387715/ARMS2 rs10490924 G→T polymorphism, and found a considerable difference in effect between wet AMD  and combined AMD . This stratification found no between-study heterogeneity for either OR1  or OR2  for combined AMD, and found a moderate degree of between- study heterogeneity for OR1  and non-significant between-study heterogeneity for OR2  of the wet AMD  and corrected p=0.669 (corrected Begg’s z=0.43), respectively. LOC387715/ARMS2 rs10490924 with G→T polymorphism; they remained significant and relatively unchanged after the third study.There was no evidence of small study bias or publication bias for the two comparisons. For the HTRA1 rs11200638 G→A polymorphism, the LOC387715/ARMS2 rs10490924 G→T polymorphism, and susceptibility to AMD. The results of our meta-analysis suggest a strong association and a moderate codominant genetic mode of action. Our primary analysis shows that, for the HTRA1 rs11200638 G→A polymorphism, the AA homozygotes carry an 8.5 fold increased risk of AMD, and the AG heterozygous variants carry just a 2.5 fold increase in risk when compared with GG homozygotes; for the LOC387715/ARMS2 rs10490924 G→T polymorphism, the TT homozygotes carry a 7.5 fold increased risk of AMD, and the TG heterozygous variants carry just a 2.4 fold increase in risk when compared with the GG homozygotes. In addition, our allele-based analysis suggests a nearly 3.0-fold increase in susceptibility to AMD among persons with the A allele of the HTRA1 rs11200638 G→A polymorphism and the T allele of the LOC387715/ARMS2 rs10490924 G→T polymorphism.To our knowledge, this is the first general overview of the association between the HTRA1 rs11200638 G→A polymorphism and AMD. When HWE was examined, 11 of the 13 studies showed no deviation and two showed some deviation. The removal of the two HW disequilibrium studies meant that our overall results were also robust; statistical adjustment for the deviations were similar and consistent with the incipient results. The point estimate values were closer to a codominant model after removal of the HW disequilibrium studies and statistical adjustment for the deviation; this suggested a multiplicative genetic mode of action that needs to be verified by more studies, particularly large-scale, long-term longitudinal studies. Moderate between-study heterogeneity was also found in the alleles (T versus G), homozygotes (TT versus GG), and heterozygotes (TG versus GG) of the association between the LOC387715/ARMS2 rs10490924 G→T polymorphism and AMD. However, the data we collected in this systematic review can only support a moderate codominant genetic model with a tight confidence interval.Our findings were based on several gene-association studies, which include several thousand participants and were robust in terms of all the planned and performed sensitivity analyses. We found no evidence of publication bias or small study bias by funnel plots and cumulative meta-analysis; moreover, “moderate,” “moderate,” and “low” degrees of between-study heterogeneity were found in alleles (A versus G), homozygotes (AA versus GG), and heterozygotes (AG versus GG) of the association between the HTRA1 gene encodes a member of the trypsin family of serine proteases [rs11200638 A risk allele affects susceptibility to AMD is still unclear [HTRA1 plays a detrimental role in arthritic disease through its capacity to degrade extracellular matrices (ECMs) directly and to upregulate the expression of matrix metalloproteinase, which results in ECM degradation [rs11200638 with AMD may be the enhancement of ECM degradation [HTRA1 in ocular tissues is unclear, it is reasonable to speculate that CNV may develop when the Bruch membrane is exposed to the detrimental effects of HTRA1. In vitro, higher luciferase expressions have been reported in both ARPE19 and HeLaS3 cells transfected with the HTRA1 rs1120638 risk homozygote (AA) genotype when compared to the wild-type (GG) [HTRA1 rs11200638 A risk allele may alter the affinity of transcription factors, including the adaptor-related protein complex 2 alpha and serum response factor to the HTRA1 promoter [HTRA1 rs11200638 A allele may increase AMD risk is its ability to bind to TGF-β family members and to inhibit signaling of TGF-β family proteins, such as bone morphogenetic protein 2 and bone morphogenetic protein 4), which have previously been reported to act as negative growth regulators in RPE [The roteases . The pre unclear ,135. Theradation . Yang etradation  hypothesradation . As showradation , the desradation . Althougype (GG) . It has promoter . Anothers in RPE ,137.HTRA1 rs11200638 G→A polymorphism and AMD was found, Kanda and others [rs11200638 G→A polymorphism of the HTRA1 gene did not make a major contribution to regulation of the HTRA1 gene and there is no association between HTRA1 G→A polymorphism and AMD. To verify these conclusions, they generated mammalian expression constructs carrying three different lengths of the normal HTRA1 promoter  and the mutant sequence carrying the AMD-risk allele at the single nucleotide polymorphism (SNP) rs11200638 (SNP-long and -medium), and these constructs were transfected into human embryonic kidney293 (HEK293), human-derived retinal pigment epithelial (ARPE19), and Human retinoblastoma (Y79) cells. As a result, they found that WT and variant SNPs of the HTRA1 promoter activities did not show significant differences in the luciferase reporter expression, and the WT-short promoter (not including the rs11200638 region) showed higher transcriptional activities than the others. A further quantitative analysis provided no evidence for significant change of mRNA expression between control and AMD retinas. This finding contrasts with the previous original experiment, which suggested an increase in HTRA1 expression in lymphocytes from AMD patients [Although an association between the d others  considerpatients ,127. Takc oxidase-deficient cones accumulate in the aging retina, particularly in the macular region [Localization of the LOC387715/ARMS2 protein to the mitochondrial outer membrane in transfected mammalian cells suggests intriguing mechanisms through which an A69S change may influence AMD susceptibility. Mitochondria are implicated in the pathogenesis of other age-related neurodegenerative diseases, including Alzheimer disease, Parkinson disease, and so on . Mitochor region ,142. Morr region . Photorer region ,144. Kanr region . HoweverHTRA1 gene rs11200638 G→A polymorphism is reasonably common, with an allele frequency of over 30% in a control population and over 40% in an Asian control population, and the T allele frequency of the LOC387715/ARMS2 rs10490924 G→T polymorphism was 25.17% in a control population and 38.67% in Asians. This means that the effect at the population level, especially for Asian populations, could be quite important. The proportion of AA and AG genotypes of the HTRA1 rs11200638 G→A polymorphism in a control population is 48% and the pooled OR for these two genotypes is 3.13. These two data were 64.07, 3.47 and 39.81, 3.07 for Asians and Caucasians, respectively. For the LOC387715/ARMS2 rs10490924 G→T polymorphism, the proportion of TT and GT genotypes in a control population is 38.89% and the pooled OR for these two genotypes is 3.05. These two data were 64.00, 3.17 and 35.40, 3.13 for Asians and Caucasians, respectively.Even though the results presented here are contradictory, the A allele of the HTRA1 rs11200638 G→A polymorphism is 56.0% . The PAR for the combined genotypes TT and GT of LOC387715/ARMS2 gene rs10490924 G→T polymorphism is 47.9% . In other words, the HTRA1 rs11200638 G→A polymorphism is involved in over half of all cases of AMD, quite close to the previous estimate of the first major AMD-susceptibility allele, CFH Y402H (58.9%) [LOC387715/ARMS2 rs10490924 G→T polymorphism is also involved in nearly half of all AMD cases. Higher PAR can explain part of why both these genes (HTRA1 rs11200638 G→A polymorphism and LOC387715/ARMS2 rs10490924 G→T polymorphism) play important roles in AMD, especially for wet AMD populations.The PAR for the combined genotypes AA and AG of the  (58.9%) . The LOCHTRA1 rs11200638 G→A polymorphism, LOC387715/ARMS2 rs10490924 G→T polymorphism, and AMD, and suggests that both of these genes play important roles in this disease. Potential gene-gene and gene-environmental interactions and possible mechanisms of AMD are also summarized and discussed. Our findings suggest that these genetic variations may serve as biomarkers enabling the diagnosis of AMD in a more efficient and economical way. However, large-scale, long-term longitudinal studies are required to substantiate and strengthen this association.In conclusion, this Human Genome Epidemiology (HuGE) systematic review presents strong evidence for an association between the


Nitric oxide (NO), produced by endothelial nitric oxide synthase (eNOS), plays a key role in the regulation of vascular tone. Endothelium-derived NO exerts vasoprotective effects by suppressing platelet aggregation, leukocyte adhesion and smooth muscle cell proliferation. The E298D polymorphic variant of eNOS has been associated with myocardial infarction (MI), but data relating to this variant are divergent in Greece. Accordingly, we examined a possible association between the E298D polymorphism of the eNOS gene and MI in a subgroup of the Greek population.The study population consisted of 204 patients with a history of MI and 218 control subjects. All subjects were of Greek origin and were selected from the general population of the greater Athens area. Genotyping was performed with melting curve analysis (Lightcycler system) of polymerase chain reaction amplified products using hybridization probes.According to the univariate findings, the risk for MI in E298D TT was 2.06  versus GG+GT and 2.34  versus GG. The risk for the T allele was estimated at 1.42  as compared to G allele. Regarding the additive model, one allele increase was associated with 43% higher risk of MI  as compared to the baseline category of homozygous GG. The positive association of TT versus GG+GT with MI risk remained even after adjusting for the main study covariates. Moreover, strong evidence was found for an increased risk for MI among carriers of the TT genotype who were smokers, hypertensive and had a family history of CAD.This study indicates that E298D polymorphism of the eNOS gene seems to be associated with MI occurrence in the Greek population. It is possible that TT genotype is closely linked to the etiology of MI even after adjusting for known MI risk factors. Nitric oxide (NO), the endothelium-derived relaxing factor, is synthesized from L-arginine by at least three isoforms of the Nitric Oxide Synthase (NOS) . Nitric oxide, produced by eNOS, diffuses from the endothelium to vascular smooth muscle cells, where it increases the concentration of cyclic guanosine monophosphate (cGMP) by stimulating soluble guanylate cyclase, leading to vascular relaxation [Endothelium-derived NO plays a key role in the regulation of vascular tone and displays vasoprotective effects by scavenging superoxide radicals  and supp894T in exon 7 of the eNOS gene results in the substitution of glutamic acid (E) at codon 298 by aspartic acid (D) (E298D). This is the only known polymorphism changing the eNOS protein sequence, leading to speculation that genetic variation at this site may alter e-NOS activity or regulation and possibly leads to endothelial dysfunction and to pathogenesis of several cardiovascular diseases [The gene encoding eNOS is located on chromosome 7q35-36 and contains 26 exons that span 21 kb [The 298D variant has been associated with myocardial infarction (MI) in Japanese , German A total of 422 subjects of Greek origin were prospectively enrolled in the study. Among them, 204 patients, diagnosed with an Acute Myocardial Infarction (AMI) or Non ST Segment Elevation Myocardial Infarction (NSTEMI) were compared to 218 control subjects.The 204 patients with MI admitted to the 1st Department of Cardiology, Onassis Cardiac Surgery Center Athens, Greece from October 2007 to April 2008. Diagnosis of AMI and NSTEMI was made by chest symptoms, electrocardiographic changes and serum creatinine kinase-MB isoenzyme (CK-MB) elevations according to guidelines .The control group consisted of 218 volunteers who were selected from the general population of the greater Athens area. Controls were not eligible to participate if they had a history of MI or angina, clinical evidence of coronary artery disease (CAD), stroke, or any atherosclerotic disease in the past, based on a detailed medical history and a physical examination followed by a normal electrocardiogram.All subjects provided written informed consent and the study was approved by the hospital ethics committee. Subjects were evaluated with a detailed questionnaire that provided information on coronary risk factors, namely smoking, family history of CAD, hypertension, diabetes mellitus and hypercholesterolemia. Patients who used to smoke more than eight cigarettes per day for more than one year were considered to be smokers (current or ex-smokers). Body mass index (BMI) was calculated in all participants.MI patients and controls were matched by gender and age ± five years. Controlling for basic socio-demographic characteristics in the statistical analysis, minimized further the consequences of potential confounding by those variables.Hypertension was defined by a history of several blood pressure measurements with elevation of either systolic (> 140 mmHg) or diastolic (> 90 mmHg) blood pressure. Diabetes mellitus was defined by elevated blood glucose levels after fasting: (> 126 mg/dL) or 2 h after 75 g of oral glucose loading (> 200 mg/dL), or by anti-diabetic drug. Hypercholesterolemia was defined by elevated total serum cholesterol levels (> 200 mg/dL).5 target equivalents per reaction). PCR primers 5'-AAggCAggAgACAgTggATggA and 5'-TgCTCCAggggCACCTCAAg and the probes CCAgATgATCCCCTAgAACTC--FL  and 640-CCTTCTgCCCCCCgAgCTggTCC--PH contained in the LightMix kit eNOS  were used in combination with the LightCycler 480 Genotyping Master and 96 well plates , following the instructions of the manufacturer. The tests were run with 5 μl (0.3 μg) sample DNA in a final volume of 20 μl; control DNA  was diluted 1:4. Each PCR run contained a negative (no template) control. To confirm reproducibility, mutation analysis of 9 random samples  was performed 10 times each.Genomic DNA was extracted from peripheral blood leukocytes using standard methods . Purified DNA (concentration of 60 μg/ml) was stored at -20°C. The E298D polymorphism was determined by Real-Time Polymerase Chain Reaction analysis using the LightCycler 480 Instrument . The published probes  were fouThe probes are designed to match with the T allele, thus reporting a lower melting temperature (Tm) for the wild type G allele. Distinction of wild type, mutant and heterozygous genotype can be easily accomplished by their respective melting temperatures.After 10 min denaturation at 95°C the amplification was performed running 45 cycles 5 s at 95°C, 10 s at 60°C and 15 s at 72°C, following the instructions of the LightMix kit. The melting program included three steps: denaturalization at 95°C for 30 s, renaturation at 40°C for 1 min and then slowly raised to 80°C to allow monitoring of the decline of fluorescence generated by melting of the hybrids, as a function of temperature. Melting curves were automatically converted to fluorescence peaks with the LightCycler 480 Instrument analysis software, allowing the distinction of genotypes. The grouping software uses a curve shape-matching algorithm to identify wild type from mutant samples and cut-offs are based on variability from the wild type curve. The results were confirmed for homozygous mutant, homozygous wild type and heterozygous samples in all repeated measurements.2 for categorical data were used to assess the differences in socio-demographic and medical history factors between cases and controls. Genotype frequencies of E298D polymorphism were compared in the two groups of participants with Fisher's exact test. To study the univariate association of MI with the eNOS genotype we modeled the data through logistic regression analysis (crude odds ratios); the relevant p-values derived from Wald test for dominant and recessive models and Armitage test for trend for additive model. Subsequently, a multivariate analysis was performed to estimate the risk of MI for TT versus GT+GG after adjustment for a series of possible risk factors and confounders (adjusted odds ratios). Results were accepted as statistically significant when two-tailed p-values were less than 0.05. The SAS statistical package  was used in all analyses whereas power calculation was performed on QUANTO.Body mass index (BMI) was regarded as continuous variable, while age, gender, kind of genotype, and presence of all coronary risk factors  were considered nominal variables. Initially, t test for continuous data and χSetting minor allele frequency (MAF) at 0.3, expected odds ratio from 1.5 to 2.5 (univariate odds ratio for the relationship between TT/GG+GT and the risk for MI), alpha 0.05 and power 80%, on QUANTO, mode of inheritance seems to result in an odds ratio of at least 2.3.Table Table In the univariate analyses of the data Table , TT homoBased on the univariate findings suggesting an increased risk for TT homozygous and the idea that the recessive model of inheritance seems to be more appropriate than dominant or additive for the needs of such a research , we furtp = 0.21) [We performed a case-control study of the common E298D polymorphism of the eNOS gene. Similarly to recent studies ,13,17-20 = 0.21) .MI is a multifactorial disease. Further genetic and environmental risk factors, such as age, gender, BMI, smoking habits and family history as well as accompanying disorders like high blood pressure and hypercholesterolemia contribute significantly to the susceptibility of MI.The homozygous T allele carriers, as suggested by the univariate analysis of the present study, have an increased risk to develop MI. We performed a multivariate analysis to examine the additive influence of age, gender, BMI, smoking habits and family history to the TT genotype. The increased risk of the TT gene remains even if we add the variables smoking, hypertension and family history. BMI seems to tentatively elevate MI risk, while gender seems to be unrelated to MI. Data from a recent study confirmed that predominantly younger T allele carriers of the eNOS E298D gene polymorphism with various coronary high-risk profiles have an increased risk to suffer MI . Taking The association between eNOS E298D polymorphism and MI is strongly affected by cigarette smoking. In a previous study it was demonstrated that the variant 298D allele is particularly associated with significantly lower endothelial-dependent vasodilation in smokers, with no significant effect in non-smokers . We obseThe results of the present study strongly suggest that eNOS gene may be a risk factor for MI. Although the mechanism by which E298D polymorphism confers susceptibility to MI is not clear, several studies have described a possible role of NO in the pathogenesis of MI and suggest that endothelial NO may have an important atheroprotective role beyond its effect on vessel tone and blood pressure ,28. ThisTwo previous studies showed a significant association between the 298D allele and the risk of MI in the Greek population ,19. A thThis study adds to the gradually developing consensus that E298D polymorphism of the eNOS gene seems to be associated with MI occurrence. It is possible that TT is closely linked to the etiology of MI even after adjusting for known MI risk factors. The underlying mechanisms are still speculative but it appears that further studies are needed to explore whether E298D polymorphism is an independent risk factor or an indirect marker of different genetic and environmental factors.No case control study is immune to criticism concerning case and control selection and the present one is no exception. However, the identification in the present study of most established risk factors for MI indicates that possible selection and information biases were not important enough to challenge the validity of the results. We have no reason to believe that cases were not representative of people suffering MI in the country. However, because of the overall low frequency of TT (15 subjects with TT genotype in the control group and 27 in the case group), the results should be viewed with caution. Notably, given the expected low frequency of TT in the general population, this type of studies face practical difficulties in achieving high number of participants. Further studies could provide an insight into the role of E298D polymorphism of the eNOS gene in the MI occurrence.This study indicates that E298D polymorphism of the eNOS gene seems to be associated with MI occurrence in the Greek population. We found evidence that homozygous TT is positively related to the risk of MI and this association is independent of possible effects of other known MI risk factors. Smokers, hypertensive and those with a family history of CAD are more likely to develop MI.The authors declare that they have no competing interests.CD carried out the molecular genetic analyses, participated in the design of the study and helped to draft the manuscript. ND conceived the study, participated in the design and coordination of the study and drafted the final manuscript. OL carried out the design of primers and probes. MR and DP helped with the statistical analysis and to draft the manuscript. DVC participated in its design, coordinated the recruitment of the patients and helped to draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2350/11/133/prepub


It is important to better understand the aetiology of thrombosed external haemorrhoids (TEH) because recurrence rates are high, prophylaxis is unknown, and optimal therapy is highly debated.We conducted a questionnaire study of individuals with and without TEH. Aetiology was studied by comparison of answers to a questionnaire given to individuals with and without TEH concerning demography, history, and published aetiologic hypotheses. Participants were evaluated consecutively at our institution from March 2004 through August 2005.One hundred forty-eight individuals were enrolled, including 72 patients with TEH and 76 individuals without TEH but with alternative diagnoses, such as a screening colonoscopy or colonic polyps. Out of 38 possible aetiologic factors evaluated, 20 showed no significant bivariate correlation to TEH and were no longer traced, and 16 factors showed a significant bivariate relationship to TEH. By multivariate analysis, six independent variables were found to predict TEH correctly in 79.1% of cases: age of 46 years or younger, use of excessive physical effort, and use of dry toilet paper combined with wet cleaning methods after defaecation were associated with a significantly higher risk of developing TEH; use of bathtub, use of the shower, and genital cleaning before sleep at least once a week were associated with a significantly lower risk of developing TEH.Six hypotheses on the causes of TEH have a high probability of being correct and should be considered in future studies on aetiology, prophylaxis, and therapy of TEH. Although anorectal disorders are common -4 no datWe limited our study to single lesions using Hancock's definition of "an acute localised thrombosis which may affect the external plexus" . SynonymTo gather data about TEH, we searched the MEDLINE database (December 1958 to January 2004) using the following keywords to find hypotheses about its pathophysiology: thrombosed external haemorrhoid, acute haemorrhoidal disease, perianal thrombosis, and thrombosed haemorrhoids.One hundred eighty-seven papers were collected and consisted of reports that were therapeutic (44.4%), concerned with foreign objects (31.6%), reviews (20.8%) or case reports (3.2%). Titles and abstracts were reviewed and hard copies obtained for further examination. Additionally we reviewed journal reverence lists and standard textbooks and applied our existing knowledge of the primary publications in this area .Individuals of both genders, aged 16 - 80 years old, who entered consecutively our outpatient clinic from March 18 2004 to August 18 2005 were enrolled. Patients had been referred from general practitioners, physicians, urologists or gynaecologists for anal  and/or abdominal complaints . After proctologic assessment in the knee-chest-position , individBoth groups of participants (with and without TEH) were asked to complete our questionnaire table  that focAt each step during the analysis we tried to enhance the significance of the statistical calculations. Initially we determined which of the 38 dichotomously coded variables  might have a significant bivariate relationship with TEH. To decipher this we used Fisher's exact test with a significance level of 5%. We than computed the odds ratio with a 95% confidence interval for each variable and searched for strong correlations.A Pearson correlation coefficient above 0.36 was defined as strong. If a strong correlation between variables was found, only one of the correlating factors was retained. The factors of age, trainee, and retirement are naturally highly correlated. To prevent multivariate analysis from being biased by multi-colinearity, we decided to retain only one of these three variables. Age was considered to be the most general underlying factor and was therefore retained for further calculations. "Use of dry toilet paper exclusively" was also retained since it was determined to be a more concise fact than "Use of dry toilet paper combined with wet anal cleaning types". The strong correlation between these two variables justified the decision to keep only one for multivariate analysis.For each of the remaining factors the variance inflation factor (VIF) was computed to ensure that multi-colinearity did not affect further evaluations. Finally a stepwise logistic regression analysis was performed to determine which of the remaining variables also had multivariate significance with TEH. For these analyses we used SPSS 15.0.1.1, 2007, SPSS Inc. Chicago, IL, USA.Our research study was carried out in compliance with the Helsinki Declaration.Each patient gave written consent to participate in the study. They gave permission to take photos of their anal lesions to be presented in scientific medical journals or for medical educational purposes.One hundred forty-eight individuals were enrolled: 72 patients with TEH only and 76 patients without TEH but with alternative diagnoses such as screening colonoscopy (22), stomach ache (21), IBD (9), colonic polyp (7), gastric ulcer (4), constipation (3), liver cirrhosis (1), cholelithiasis (1), pruritus ani (2), haemorrhoids (2), fissure-in-ano (2), rectal prolaps (1) or proctitis (1). Demographic variables in both groups of individuals were homogenously distributed , excessive physical effort , and use of dry toilet paper combined with wet cleaning methods after defaecation . Three factors were found to be associated with a significantly lower risk for TEH: use of the bathtub , use of the shower , and cleaning genitals before sleep at least once a week .A second multivariate analysis was performed using female patients only (N = 58) to determine whether pregnancy ,27,28 isAs additional hypotheses regarding the aetiology of a disease are introduced, it becomes increasingly necessary to determine which risk factors are real and which are fiction. Numerous ideas about the aetiology of TEH have been published -28,32,33To better understand the risk factors for TEH, we initiated an explorative study encompassing a wide spectrum of demographic factors. One disadvantage of the study may be the small number of individuals used to gather data 148 in total). One advantage of the study is that the data was gathered prospectively unlike other available TEH studies, which are primarily retrospective analyses ,8,10,16. in totalIn females our multivariate analysis contrasted with data published by others -28 and sAdditional potential aetiologies of TEH include local irritation or inflammation of anal skin triggered by different agents like ointments ,23,28, sThere were no competing interests. The study was sponsored by the authors themselves. There were no financial or non-financial competing interests .HR had the idea. All authors contributed to the design of the study and construction of the study protocol. OG was responsible for the research of literature. OG and HR saw the patients, and asked them to tick their answers into patients' questionnaire before proctologic assessment. Findings were ticked into a PC-documentation-sheet directly after medical assessment of each patient by OG and HR. Results were discussed with all authors. YH was responsible for statistical evaluations. OG wrote the first drafts of the paper which were revised by all authors. HR wrote the final draft.


KIF6 (rs20455) was associated with coronary events in Caucasian participants of five prospective studies. We investigated whether this KIF6 variant was associated with non-fatal myocardial infarction (MI) in a case-control study of an admixed population from the Central Valley of Costa Rica. Genotypes of the KIF6 variant were determined for 4,134 men and women. Cases  had survived a first MI; controls  had no history of MI and were matched to cases by age, sex, and area of residence. We tested the association between the KIF6 719Arg allele and non-fatal MI by conditional logistic regression and adjusted for admixture of founder populations.The 719Arg allele of KIF6 719Arg carriers were not at significantly higher risk for non-fatal MI in this study after adjustment for traditional risk factors or admixture . Heterozygotes of the KIF6 Trp719Arg variant were at increased risk of non-fatal MI: the adjusted odds ratio was 1.16 , but this association would not be significant after a multiple testing correction.Compared with the reference Trp/Trp homozygotes, KIF6 719Arg allele were not at increased risk of non-fatal MI in a case-control study of Costa Ricans living in the Central Valley of Costa Rica.We found that carriers of the  KIF6 Trp719Arg was not associated with coronary artery disease (CAD) in two recently reported case-control studies: the Ottawa Heart Genomics studyKinesins are a superfamily of homodimeric motor proteins that transport cellular cargos  along microtubules in an ATP dependent processKIF6 719Arg allele is associated with CHD in other ethnic populations. For example, among African Americans in ARIC, each allele of KIF6 719Arg increased the risk for incident CHD by 1.23-fold KIF6 719Arg allele had a hazard ratio for incident myocardial infarction (MI) of 4.14 , compared with noncarriersKIF6 719Arg allele and CHD were not statistically significant in the small African American study populations in ARIC and CHS, these results suggested that the KIF6 719Arg allele may be associated with CHD in ethnic populations other than Caucasians. Therefore, we asked whether the KIF6 719Arg allele is associated with MI in a Hispanic population from Costa Rica.Only limited data are available regarding whether the i.e., matched characteristics) did not differ significantly between cases and controls (KIF6 SNP (rs20455) that cause a Trp719Arg variation did not deviate from the distribution expected under Hardy–Weinberg equilibrium (P = 0.38); the frequency of the minor allele (KIF6 719Arg) was 0.36 in the control subjects.The characteristics of cases with myocardial infarction and population-based controls from the Central Valley of Costa Rica used in this study are shown in controls . Other Ccontrols . The genKIF6 719Arg variant were not at increased risk of MI , compared with major allele homozygotes (Trp/Trp), was 1.15  were not at increased risk of MI, compared with major allele homozygotes (Trp/Trp).We found that carriers of one or two copies of the KIF6 719Arg carriers were not at significantly higher risk for non-fatal MI in this case-control study of a Hispanic population from Costa Rica after adjustment for traditional risk factors or admixture. Heterozygotes of the KIF6 Trp719Arg variant, compared with Trp/Trp homozygotes, were at increased risk of nonfatal MI (OR = 1.16), but this association would not be significant after a multiple testing correction. Arg/Arg homozygotes of the KIF6 variant were not at increased risk of MI, compared with Trp/Trp homozygotes.In contrast to the results reported from 5 prospective studies, we found that KIF6 variant has been found to be associated with CHD in five prospective studies of CHDKIF6 719Arg and MI in a population with angiographically defined coronary artery disease. The Wellcome Trust Case-Control Consortium analyzed the association between KIF6 Trp719Arg and CAD. That KIF6 719Arg was associated with CHD in prospective, but not in case-control studies, could be explained by either ascertainment biases or by statin use in the populations from which the cases were drawn. Statin use would be expected to reduce the likelihood of observing an association between KIF6 719Arg and CHD because in 3 randomized studiesKIF6 719Arg carriers but not in noncarriers. The potential inclusion of individuals on statin therapy is also a limitation of our Costa Rica study, which may have weakened the association of the KIF6 variant with MI.Although the KIF6 719Arg are consistent with the pattern of these previous studies. The only prospective study where the observed risk estimates have been higher for Arg/Arg homozygotes than heterozygotes of KIF6 Trp719Arg was the ARIC studyIn previous published prospective studies, risk estimates have been nominally higher for heterozygotes (Trp/Arg) than for homozygotes (Arg/Arg) in most studies of Trp719Arg KIF6 719Arg and MI. Based on historical recordsKIF6 (719Arg) allele frequency of the KIF6 Trp719Arg variant reported in the HapMap database differs between these populations. The allele frequencies are 38 percent for Caucasians of Western European decent and 95 percent for West Africans ; the allele frequency for Amerindians from the central valley of Costa Rica was not available from HapMap. Since the KIF6 719Arg allele frequency varies according to ancestry, a spurious association could occur if the admixture of the 3 ancestral populations were different between cases and controls. However, adjustment for the degree of admixture did not appreciably change the KIF6 719Arg risk estimate for MI.Admixture in the Costa Rican population did not affect the association between KIF6 719Arg in CARE and WOSCOPSSince participants with cardiovascular diseases other than a history of MI were not excluded from the control group, the risk estimates obtained in this study may be lower than would have been observed had a disease-free control group been used. The use of statins was previously reported to ameliorate the risk of coronary events in carriers of KIF6 719Arg allele were not at increased risk of non-fatal MI, compared with Trp/Trp homozygotes, in a case-control study from the Central Valley of Costa Rica. We found no evidence that admixture within the Costa Rican population affected these results. Additional studies are need to address why the association between carriers of the KIF6 719Arg and MI differ between this and other case-control studies and the previously reported prospective studies.Carriers of the 2 and 2,057,000 culturally Hispanic individuals living in Costa Rica. This area included 36 counties in the Central Valley of Costa Rica representing a full range of socioeconomic levels, as well as urban, periurban, and rural lifestyles. Medical services for all residents of this area were provided by 6 large hospitals, which are part of the National Social Security System. Eligible case participants were men and women who were survivors of a first MI as diagnosed by a cardiologist at any of the 6 recruiting hospitals in the catchment area between 1994 and 2004. Since this study was designed to draw cases and controls who were representative of the general population and the intent was that cases should differ from controls only by the case definition, the presence of other cardiovascular conditions was not used to exclude controls (or cases). Control participants were randomly selected using data from the National Census and Statistics Bureau of Costa Rica. Control participants were ineligible if they had ever had an MI. The controls in this study were matched to cases by assigning participants to strata based on age groups (±5 y), sex, and area of residence (county). Four controls were excluded from the study because they were assigned to strata for which there were no matching cases, resulting in 1987 cases and 2147 controls. Study and consent documents were approved by the Human Subjects Committee of both the Harvard School of Public Health and the University of Costa Rica—all participants gave written informed consent.The design of the original population-based case-control study has been described in detail elsewhereKIF6 Trp719Arg genotypes were determined for all study participants using an allele–specific real-time PCR genotyping assayThe admixture in this study population was previously determined using the program ADMIXMAP and 39 AIMs to estimate the proportion of admixture of the 3 ancestral ethnicities for each subject2 test (for discrete variables). Deviations from Hardy–Weinberg expectations were assessed using an exact test in controls. Conditional logistic regression models were used to assess the association between genotype and risk of MI while taking into account the matching for age group, sex, and area of residence. SAS version 9 software was used for all regression models. Models were adjusted for smoking status (current vs. past or never smoked)P-values are 2-sided.Differences between baseline characteristics of patients were assessed by the t-test (for continuous variables) or by the χWe are grateful to the study participants, to Xinia Siles for data collection and study management in Costa Rica, and to the staff of Proyecto Salud Coronaria, San José, Costa Rica. We also thank the investigators who participated in the collection of the clinical samples and Nam Bui, David Ross, and David Wolfson for expert technical support.Table S139 Ancestry Informative Markers(0.09 MB DOC)Click here for additional data file.


Although total ankle replacement (TAR) is a recognized procedure for treatment of the painful arthritic ankle, the best choice of implant and the long-term results are still unknown. We evaluated the survival of two TAR designs and factors associated with survival using data from the nationwide arthroplasty registry in Finland.573 primary TARs were performed during the period 1982–2006 because of rheumatic, arthritic, or posttraumatic ankle degeneration. We selected contemporary TAR designs that were each used in more than 40 operations, including the S.T.A.R. (n = 217) and AES (n = 298), to assess their respective survival rates. The mean age of the patients was 55 (17–86) years and 63% of operations were performed in women. Kaplan-Meier analysis and the Cox regression model were used for survival analysis. The effects of age, sex, diagnosis, and hospital volume were also studied.5 inhabitants. The 5-year overall survivorship for the whole TAR cohort was 83% (95% CI: 81–86), which agrees with earlier reports. The most frequent reasons for revision were aseptic loosening of one or both of the prosthesis components (39%) and instability (39%). We found no difference in survival rate between the S.T.A.R. and AES designs. Furthermore, age, sex, diagnosis, and hospital volume  did not affect the TAR survival.The annual incidence of TAR was 1.5 per 10Based on our findings, we cannot conclude that any prosthesis was superior to any other. A high number of technical errors in primary TARs suggests that this low-volume field of implant arthroplasty should be centralized to fewer units. Most TA5 .We analyzed the survival of two TAR designs and the factors affecting survival at a national level by using the data from the Finnish Arthroplasty Register.Our study was based on information recorded in the Finnish Arthroplasty Register  relatingThere were data on 645 TARs, each of which have been recorded individually for every operation since the start of the register. Of these 645 TARs, 573 (89%) were primary operations and 72 (11%) were revisions.In order to assess the survival of different TAR designs, we selected only those designs that had been used in more than 40 operations during the study period . In addiTo meet our inclusion criteria, the following implants were selected: LINK S.T.A.R.  System  and BIOMET AES (Ankle Evolutive System) . Both prostheses are third-generation mobile-bearing designs. All the S.T.A.R. prostheses were uncemented, double-coated versions. All the AES implants were uncemented, but the implant was changed by the manufacturer in 2004—from single-coated to a porous double-coated version with monobloc tibial component (the earlier tibia was modular). Of the other implant designs in use in Finland at the end of 2006, the Hintegra  was excluded due to an insufficient number of operations (11). The use of the earlier designs was infrequent, and the designs were discontinued in the late 1980s . 515 priHalf of the 515 TARs were performed due to rheumatoid arthritis (RA) . Other indications included posttraumatic  and primary  osteoarthritis, other arthritides , and other diseases . We analyzed the overall survival of TARs performed due to RA and compared it to the “other indications” group in order to assess the effect of underlying disease.355 (69%) of the 515 TARs were performed in 1 foundation-based hospital and 2 university hospitals, each of which performed more than 100 TARs during the period 1997–2006 (group A). 4 more hospitals had performed 10–50 TARs (group B), and 10 other hospitals less than 10 operations (group C). We analyzed the overall survival of TARs performed in high-volume hospitals (group A) and compared this to that of the 2 low-volume hospital groups together (A vs. B and C) and separately (A vs. B vs. C) in order to assess the effect of hospital volume.The endpoint for survival was defined as revision with either one component or the whole implant removed or exchanged. Kaplan-Meier survival data were used to construct the survival probabilities of implants at 1, 3, 5, and 7 years. Survival data obtained in the Kaplan-Meier analysis were compared by the log-rank test. The Cox multiple-regression model was used to study differences between groups and to adjust for potential confounding factors. In all models, the confounding factors were age and sex. The factors studied with the Cox model were TAR design and hospital type . All models included adjustment for differences in age and sex.The Cox regression analyses provided estimates of survival probabilities and revision risk ratios (RR) for different factors. Estimates from the Cox analyses were used to construct adjusted survival curves at mean values of the risk factors. The Wald test was used to calculate p-values for data obtained from the Cox multiple-regression analysis. Differences between groups were considered statistically significant if the p-values were less than 0.05 in a two-tailed test. We used the SPSS software version 17.0.5 inhabitants (463 (90%) of the primary TARs were unilateral, and in 26 patients a 2-stage bilateral replacement was done. Of the 515 TAR operations, 327 (63%) were performed in women. At the time of the operation, the mean age of the patients was 55 (17–86) years. During the last 5 years, the mean annual incidence of TAR was 1.5 (95% CI: 1.3–1.8) per 10abitants .Over the whole study period, 9 TAR designs were used but only 2 of them had more than 40 operations . At the The Cox regression model (adjusted for age and sex) showed a similar risk of revision in all disease subgroups.We found no differences in survival rates between S.T.A.R. and AES TAR designs over the whole study period, using either Kaplan-Meier analysis or the Cox regression model (with or without adjustment for age and sex)  and 3. TAge and sex did not have any statistically significant effect on survivorship in the Cox multiple regression models (unadjusted and adjusted for implant design).Hospital volume did not have any statistically significant effect on survivorship in the Cox multiple regression models (unadjusted and adjusted for implant design).During the period 1997–2006, 59 revisions were reported . Thus, tOur main finding was the relatively low survival of TAR, with only 83% of cases revision-free at 5 years. We found no differences in survival between different TAR designs or concepts. Age, sex, diagnosis, and hospital volume did not affect the TAR survival. The mean annual incidence of TAR was 1.5 per 105, which is twice the reported incidence in Sweden .We acknowledge that our registry-based study has certain limitations. For example, we were not able to report any subjective outcome measurements, e.g. ankle performance scores or disease-specific quality of life measurements. Moreover, no radiographic analyses were done. Furthermore, in rheumatoid patients, a registry-based study may have pitfalls in that some of the patients diagnosed with RA are actually affected by juvenile or other subtypes of chronic arthritis. Another important issue, especially with an ankle register, is that a failed TAR is often converted to an arthrodesis. This may result in under-reporting of failures despite instructions, because the implants used in ankle arthrodesis are not reported to the arthroplasty register and a small number of these conversions may be done by orthopedic surgeons who are not familiar with reporting to the register.In studies based on the Swedish and Norwegian national registries, aseptic loosening has been the most frequent reason for revision (31–48%)  compared to the Swedish (16%) and Norwegian results (14%), which probably indicates a long learning curve for TAR. However, some of the primary TARs revised for instability may have been in acceptable alignment and balance immediately after surgery. In these cases, a marked preoperative varus or valgus deformity may have been corrected but degenerated ligaments may have been insufficient to sustain the balance after full weight bearing, and components may have migrated during follow-up due to poor bone quality, especially in RA. The migrated prosthesis components were found to be well-fixed in many of these cases at the time of revision.It is commonly believed that the patient’s age, sex, and diagnosis have an influence on implant survival in TAR. In the Swedish study, only lower age was associated with an increased risk of later revision, but similarly to the Norwegian study we did not find such an association.There was an obvious shift from the S.T.A.R. prosthesis to the AES prosthesis during 2002–2003. The main reasons for this are technical: surgical instrumentation for the S.T.A.R. prosthesis is more difficult to use and the lack of trial implants caused many ankle surgeons to switch to the AES . Also, tBased on our findings, we cannot conclude that any prosthesis was superior to any other. A high number of complications in primary TARs were associated with balance and instability. These issues require a profound understanding of not only the ankle joint, but also the whole hindfoot. Based on earlier studies on low-volume joint replacement , we sugg


Previous evidence implicates polymorphisms of GSTM1 and GSTT1, candidates of phase II enzymes, as risk factors for various cancers. A number of studies have conducted on the association of GSTM1 and GSTT1 polymorphism with susceptibility to nasopharyngeal carcinoma (NPC). However, inconsistent and inconclusive results have been obtained. In the present study, we aimed to assess the possible associations of NPC risk with GSTM1 and GSTM1 null genotype, respectively.The associated literature was acquired through deliberate searching and selected based on the established inclusion criteria for publications, then the extracted data were further analyzed using systematic meta-analyses.A total of 85 articles were identified, of which eight case-control studies concerning NPC were selected. The results showed that the overall OR was 1.42 (95%CI = 1.21–1.66) for GSTM1 polymorphism. While for GSTT1 polymorphism, the overall OR was 1.12 (95% CI = 0.93–1.34).The data were proven stable via sensitivity analyses. The results suggest GSTM1 deletion as a risk factor for NPC and failed to suggest a marked correlation of GSTT1 polymorphisms with NPC risk. Nasopharyngeal cancer (NPC), a fast-growing tumour, characterized by a high frequency of nodal and distant metastasis at diagnosis, is rare in many areas of the world but common in Southeast Asia . EvidencXenobiotics can be detoxified by phase II enzymes, such as GSTM1 and GSTT1 which have been suggested to be involved in detoxification of polycyclic aromatic hydrocarbons (PAHs) and benzo(a)pyrene . EvidencSince a single study may have been underpowered to clarify the associations of GSTM1 or GSTT1 polymorphisms with NPC susceptibility, in the present study we aimed to perform evidence-based quantitative meta-analyses that might increase statistical power to address this controversy.GSTM1, GSTT1, nasopharynx or nasopharyngeal, head and neck, cancer, carcinoma, tumour or neoplasm.We carried out a search in the Medline, EMBASE and Chinese National Knowledge Infrastructure (CNKI) without a language limitation, covering all papers published up to Sep 2008, with a combination of the following keywords: We evaluated potentially associated publications by checking their titles and abstracts and then procured the most relevant publications for a closer examination. Moreover, the reference lists of the selected papers were also screened for other potential articles that possibly have been missed in the initial search. The following criteria were used for the literature selection of the meta-analysis:1. Articles clearly describing studies in the association of NPC with GSTM1 or GSTT1 polymorphisms;2. Case–control studies;3. The NPC diagnoses and the sources of cases and controls should be stated;4. The size of the sample, odds ratios (ORs) and their 95% confidence intervals (CIs) or the information that can help infer the results should also be offered;5. Those publications that presented data allowing such outcomes to be derived were also selected.Accordingly, the following exclusion criteria were also used:1. Design and the definition of the experiments were obviously different from those of the selected papers;2. The source of cases and controls and other essential information was not offered;3. Reviews and repeated literature.After searching, we reviewed all papers in accordance with the criteria defined above for further analysis. In addition, Hardy-Weinberg equilibrium test  was condData were extracted and entered into a database. The extraction was performed by two reviewers independently. For conflicting evaluations, an agreement was reached following a discussion.The odds ratio (OR) of GSTM1 or GSTT1 polymorphisms and NPC risk was estimated for each study. For detection of any possible sample size biases, the OR and its 95% confidence interval (CI) to each study was plotted against the number of participants respectively. A Chi-square based Q statistic test was performed to assess heterogeneity. If the result of the heterogeneity test was P > 0.05, ORs were pooled according to the fixed-effect model (Mantel-Haenszel), Otherwise, the random-effect model (DerSimonian and laird) was used. The significance of the pooled ORs was determined by Z-test. The Hardy-Weinberg equilibrium was assessed via Fisher's exact test.Publication bias was assessed by fail-safe number for P = 0.05 (Nfs0.05) . StatistA total of 85 studies regarding GSTM1 or GSTT1 were identified  and 463 cases and 803 controls had the GSTM1 present genotype. The overall OR was 1.42 (95% CI = 1.21–1.66) and the test for overall effect Z value was 4.39 (P < 0.05). The results indicate that GSTM1 null genotype might have an association with increased risks of NPC.For GSTT1 polymorphism, the data available for our meta-analysis were obtained from 4 case-control studies of 790 cases and 1156 controls, of which 385 cases and 518 controls had the null genotypes (the exposure group) and 405 cases and 638 controls had the present genotype of the GSTT1 gene. As shown in Fig. Considering that the study  concerniIn order to compare the difference and evaluate the sensitivity of the meta-analyses, we also reported the results of the random-effect model for GSTM1 as follows: the combined OR and 95% CI were 1.42 (95% CI = 1.21–1.66), similar to the results of the fixed-effect models. For GSTT1, the results of the fixed-effect model and random effect model were statistically similar, as stated in the above section.Additionally, we also conducted one-way sensitivity analysis  to evaluFunnel plots were usually created to assess the possible publication biases. In the meta-analyses, for GSTT1 and GSTM1 polymorphisms, the funnel plots were not created because it is useless when the number of the included studies is limited. Nevertheless, fail-safe number, for the evaluation of the reliability of meta-analysis, is defined as the number of negative results that could reverse the significant finding. The Nfs0.05 for GSTM1 polymorphism was 66, suggesting that the publication biases might not have a remarkable influence on the results of the meta-analyses. Notably, for GSTT1 polymorphism, it is useless to utilize fail-safe number for evaluation of publication bias when the number of the included studies is only four.Previous evidence suggests that GSTM1 and GSTT1 polymorphisms may have a close association with increased susceptibility to various carcinomas. In the present study, the results of meta-analyses suggest that genetic deletion of GSTM1 may contribute to increased susceptibility to NPC whereas GSTT1 polymorphism may not.Null mutations of GSTM1, one of the most important phase II enzymes, are known to abolish enzyme activities and therefore have been linked with increasing incidence of certain cancers, most likely due to increased susceptibilities to environmental toxins and carcinogens. Previous meta-analyses indicate that GSTM1 deficiency might have a significant association with increased risks of breast cancer  and lungSimilarly, null genotype of GSTT1 has been suggested to associate with risks of a number of cancers. Previous meta-analyses suggest marked associations of GSTT1 deletion with lung cancer , gastricIn this meta-analysis, we did not perform subgroup analyses. First, for GSTM1 polymorphisms, the data showed an absence of heterogeneity between the included studies. In addition, the extracted data showed that most studies were conducted on Asians. Of the eight studies, only a French study concerned Caucasian while another American study concerned a combined population with several ethnicities. Hence, a subgroup analysis regarding ethnic stratification had not been performed. Second, for GSTT1 deletion, we excluded the French study that might be different from the other three studies. As a consequence, the data failed to show a significant association of GSTT1 null genotype with NPC risk in Asians Fig.  or in thThe results of the present meta-analysis indicated a possible role of GSTM1 deletion in the tumorigenesis and progression of NPC. Nevertheless, the data failed to show a significant association of GSTT1 null genotype with increased susceptibility to NPC. This discrepancy might be due to some reasons. For GSTT1, a gene that is highly conserved during evolution, major ethnic differences exist in frequency distribution. In East Asia, highest percentages of individuals with the GSTT1 null genotype were reported . InteresSeveral limitations might be included in this study. Since most of the included studies have conducted on Asians and a few on Caucasians, the results must be interpreted with caution. Further studies concerning populations in other areas such as African and American are required to diminish the ethnic variation-produced biases. Additionally, a possible publication bias might have been introduced as only published studies written in English and Chinese as well as French that could be searched from Medline database were included. Notably, we did not use the funnel plots and Egger's linear regression test  for asseIn conclusion, the data of the present meta-analyses indicate GSTM1 polymorphism as a risk factor for NPC and failed to show a significant association of GSTT1 polymorphism with NPC risk.The authors declare that they have no competing interests.XZ and LC conceived of the study, and carried out the analysis of the literatures and drafted the manuscript. ZX carried out the collection of the literatures. QL helped with the statistical analysis and manuscript drafting. XZ conceived of the study, and participated in its design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.


These inconclusive results motivated us to further examine the relationship between CYP1A1 MspI and exon 7 polymorphisms and risk of oesophageal cancer. In total, 146 cases of oesophageal squamous-cell-carcinoma and 324 control cases  were genotyped from records at three Taiwan hospitals. No significant association was noted for the CYP1A1 MspI polymorphism variable between carcinoma and control cases. In contrast, the frequency of Ile/Ile, Ile/Val, and Val/Val in exon 7 was 68 (46.6%), 62 (42.5%), and 16 (11.0%) in carcinoma cases and 179 (55.3%), 127 (39.2%), and 18 (5.6%) in control cases, respectively. After factoring out other potential contributing factors, patients with Val/Val showed a 2.48 (95% CT=1.15–5.34) greater risk of developing oesophageal cancer than those with Ile/Ile. A slightly  greater risk was identified in subjects with Ile/Val . These findings suggest that an exon 7 polymorphism, not a MspI polymorphism, in CYP1A1 may be pivotal in the development of oesophageal cancer.Several British Journal of Cancer (2002) 87, 529–532. doi:10.1038/sj.bjc.6600499www.bjcancer.comCancer Research UK© 2002  CYP1A1 gene code in the aryl hydrocarbon hydroxylase (AHH) enzyme is closely associated with the metabolism of polycyclic aromatic hydrocarbons (PAHs) carcinogens  from Linxian, a region of high oesophageal cancer risk in China. Recently, CYP1A1 Val/Val genotype had a higher risk of developing oesophageal cancer than those with Ile/Ile . In view of the apparently significant influence of ethnicity in previous studies, we have examined further the role of CYP1A1MspI and exon 7 polymorphisms in oesophageal cancer risk in Taiwan.The n=324). Healthy control subjects were selected based on being the same gender as the recruited carcinoma patient and being in the hospital during the same time and of roughly the same age (±3 years). Subjects in this study were interviewed by trained interviewers using a standardized questionnaire to collect demographic and substance use  information. This study was approved by National Taiwan University's IRB Hospital. Informed consent was obtained from all subjects.Over the four-year period of this study, we recruited 146 patients with pathologically-proven oesophageal squamous-cell-carcinoma undergoing treatment at three hospitals in Taiwan, including the National Taiwan University Hospital, the Kaohsiung Medical University Hospital, and the Kaohsiung Veterans General Hospital. Concurrently, the preventive medicine department at each hospital did their best to also recruit 1–4 malignancy-free subjects per recruited carcinoma patient as healthy controls . When an MspI restriction site was present, the fragment of 340 bp was digested into two lengths of 140- and 200-bp. Homozygous wild-type individuals lacked the 140- and 200-bp fragment, and heterozygous individuals had three bands; homozygous-rare allele individuals lacked the large parent band while showing the smaller bands.The 10-μl amplification result was digested using 3 units BsrDI polymorphism in CYP1A1 exon 7 was determined using PCR and RFLP, according to the method used by BsrDI. When a BsrDI restriction site was present, the fragment of 204-bp was digested into two lengths of 65- and 139-bp. Individuals with Ile/Ile had the 65- and 139-bp fragment, heterozygous individuals had three bands; and individuals with Val/Val retained the larger parent bands (204-bp).The We included one positive control and one negative control samples in each genotyping set (∼10 samples). The positive control sample was included to ensure complete digestion of the PCR product by restriction enzymes. The negative control was placed with the same reagents as those used with actual samples, with the exception of DNA templates.CYP1A1 MspI and exon 7 polymorphisms and oesophageal cancer risk. Factors of influence (covariates) in the models included the variables of age (>65 years and ⩽65 years), gender, education level , and race . The data were analysed using the SAS statistical package and all P-values were two-sided.The gene frequency and Hardy-Weinberg equilibrium tests were conducted, with results entered into multiple logistic regression models to determine, after adjustments had been made for other factors of influence, the level of association between P=0.03), alcohol (P=0.03), and areca (P<0.01) provided the most significant predictors of oesophageal cancer risk , 146 (45.1%), and 42 (13.0%), respectively. The gene frequency of the variant allele was 35.5%. The frequency of Ile/Ile, Ile/Val, and Val/Val in exon 7 among controls was 179 (55.3%), 127 (39.2%), and 18 (5.6%), respectively. The gene frequency of the variant allele  was 25.2%. The distribution of the different genotypes in both MspI and exon 7 polymorphisms among the 324 control cases closely conformed to Hardy-Weinberg expected frequencies .The frequency of Ile/Ile, Ile/Val, and Val/Val in exon 7 was 68 (46.6%), 62 (42.5%), and 16 (11.0%) in carcinoma cases and 179 (55.3%), 127 (39.2%), and 18 (5.6%) in control cases, respectively. As lists of those genotypes with elevated oesophageal cancer risk were similar both after adjusting only for substance use and after adjusting for substance use along with age, gender, education, and race, we present here the results for the latter category. Compared to those with Ile/Ile, subjects carrying Val/Val had a 2.34-fold risk (95% CI=1.13–4.85) of developing oesophageal cancer. After adjusting for other potential confounding factors, the result remained similar  . A slight, but not significant, risk elevation was found in subjects with Ile/Val both before and after adjusting for confounding factors . In contrast, we found no significant polymorphic effect of CYP1A1 MspI on oesophageal cancer  of developing oesophageal cancer than those with the Ile/Ile genotype. In contrast, the CYP1A1 MspI genetic polymorphism was not found to be associated with elevated oesophageal cancer risk. Several studies on ethnic Japanese populations reported no association between CYP1A1MspI and exon 7 polymorphisms and risk of oesophageal cancer . Our findings approximate the The results of this study found that those cases with the CYP1A1 gene might contribute to oesophageal cancer development. In two in vitro studies, CYP1A1MspI and exon 7 polymorphisms in peripheral lymphocytes in 84 healthy Japanese male subjects in vitro. They found the age-adjusted AHH inducibility (mean±standard error) of the wild-type, heterozygous, and homozygous variants of the CYP1A1 MspI gene to be 4.89±0.36, 4.82±0.29, and 13.61±1.44, respectively. Furthermore, the homozygous variants had significantly higher AHH inducibility than the combined wild-type and heterozygous variants. However, no association was found between AHH inducibility and CYP1A1 exon 7 polymorphisms. In contrast, MspI and exon 7 polymorphisms in mitogen-stimulated lymphocytes in 51 healthy subjects. They reported that subjects with the exon 7 polymorphism  (n=12) had a relative level of CYP1A1 mRNA inducibility  of 9.0±1.7, versus 5.9±0.6 in people with the wild-type alleles (n=39). However, variant genotypes at the MspI site had no effect on CYP1A1 gene induction. Our results in this epidemiologic study align more closely with those obtained by Human cancers, e.g., gastrointestinal cancers, were suggested to be the result of the activation of procarcinogens into carcinogens (CYP1A1 MspI polymorphisms to be 42.5% (34), 42.5% (34), and 15.0% (12), respectively, among 80 coke-oven workers  of homozygous wild-types, heterozygous variants, and homozygous variants in CYP1A1 exon 7 (but not MspI) polymorphism to be a factor in elevated oesophageal cancer risk. This association suggests that carcinogen biotransformation may be a contributing factor to the etiology of oesophageal squamous-cell-carcinoma among the population in Taiwan.In summary, this study found


PEDF) Met72Thr gene polymorphism (rs1136287) was identified as a novel risk factor for exudative AMD in Chinese patients from Taiwan. The role of this polymorphism, however, has not yet been determined in a white European population. In addition, two other PEDF gene polymorphisms, −5736T>C (rs12150053) and −5304C>T (rs12948385), have been associated with increased risk of diabetic retinopathy, but have not yet been studied among patients with exudative AMD. The purpose of the present study was thus to investigate a hypothesized association between these PEDF polymorphisms and the presence of exudative AMD in a white European population.Exudative age-related macular degeneration (exudative AMD) is a common vision-threatening disease, with both environmental and genetic factors contributing to its development. Recently, homozygosity for the 72Met variant of the pigment epithelium-derived factor (PEDF polymorphisms were determined by 5′-exonuclease assays (TaqMan).The present case-control study comprised 269 patients with exudative AMD and 155 control subjects. Genotypes of the PEDF genotype and allele frequencies were not significantly different between AMD patients and control subjects. The two promoter polymorphisms, −5736T>C (rs12150053) and −5304C>T (rs12948385), were in complete association. Presence of the homozygous PEDF 72 Met/Met genotype was associated with a nonsignificant odds ratio of 1.00 . Similarly, presence of the homozygous PEDF −5736 TT genotype or −5304 CC genotype was associated with a nonsignificant odds ratio of 0.99 . Both promoter polymorphisms were in linkage disequilibrium with the Met72Thr (rs1136287) polymorphism (D'=0.83) and formed three common and one rare haplotype. Haplotype frequencies were similar between AMD patients and control subjects (p>0.05).PEDF polymorphisms is likely a major risk factor for exudative AMD in a white European population.Our data suggest that none of the investigated  Exudative age-related macular degeneration (exudative AMD) is a major cause of severe visual impairment in patients older than 50 years –3. An imPigment epithelium-derived factor (PEDF), a 50 kDa glycoprotein belonging to the serine proteinase inhibitor family –10, is aPEDF gene polymorphism, which is characterized by a methionine to threonine substitution at amino acid position 72 of PEDF   and −5304C>T [rs12948385]) we investigated do not affect any of these putative binding sites. The Met72Thr polymorphism lies at the end of a helix domain of the PEDF protein and results in the exchange of a hydrophobic amino acid (methionine) by a polar amino acid (threonine) [Currently, the functionality of the promoter , but thereonine) . NeverthPEDF in the chorioretinal tissue may modulate the efficacy as well as frequency of anti-vascular endothelial growth factor treatment. Thus, further studies focusing on the identification of functional PEDF polymorphisms and their potential association with treatment outcome might be of great interest.Previous studies have clearly demonstrated that PEDF itself is able to inhibit the development of choroidal neovascularizations –21. Cons


Despite the recent success of genome-wide association studies in identifying novel loci contributing effects to complex human traits, such as type 2 diabetes and obesity, much of the genetic component of variation in these phenotypes remains unexplained. One way to improving power to detect further novel loci is through meta-analysis of studies from the same population, increasing the sample size over any individual study. Although statistical software analysis packages incorporate routines for meta-analysis, they are ill equipped to meet the challenges of the scale and complexity of data generated in genome-wide association studies.We have developed flexible, open-source software for the meta-analysis of genome-wide association studies. The software incorporates a variety of error trapping facilities, and provides a range of meta-analysis summary statistics. The software is distributed with scripts that allow simple formatting of files containing the results of each association study and generate graphical summaries of genome-wide meta-analysis results.http://www.well.ox.ac.uk/GWAMA.The GWAMA  software has been developed to perform meta-analysis of summary statistics generated from genome-wide association studies of dichotomous phenotypes or quantitative traits. Software with source files, documentation and example data files are freely available online at  Genome-wide association (GWA) studies of hundreds of thousands of single nucleotide polymorphisms (SNPs), genotyped in samples of thousands of individuals, such as those undertaken by the Wellcome Trust Case Control Consortium , have prMeta-analysis of GWA studies has been greatly assisted by the development of imputation techniques ,6 which The statistical methodology underlying meta-analysis is already well established , and freTo address these challenges, we have developed the GWAMA  software to perform meta-analysis of summary statistics generated from GWA studies of dichotomous phenotypes or quantitative traits. The software incorporates tools to align studies to the same reference strand, irrespective of the GWA genotyping product, where possible, and optionally performs genomic control  of summaN GWA studies, not necessarily typed using the same genotyping product or imputed to the same reference panel. We assume that studies have been filtered for appropriate quality control metrics to exclude poorly genotyped or imputed SNPs -1 is the inverse of the variance of the estimated allelic effect in the ith study, obtained from the standard error . Note that if the jth SNP has not been directly genotyped or imputed as part of the ith study, wij = 0. The variance of the combined allelic effect across studies is given by 2 distribution with one degree of freedom, and this provides the basis of a test of association of the trait with the jth SNP over all studies.where λDi and λD*i, respectively, for the ith study [K is replaced by D or D*, as appropriate. Furthermore, users have the option of correcting for between-study variation across the meta-analysis so that λ is the genomic control inflation factor over all meta-analysed association test statistics, genome-wide.The presence of population structure in a GWA study, not taken account of in the analysis, can lead to over-dispersion of the corresponding association test statistics. One approach to combat this problem is to correct test statistics by the genomic control inflation factor. This factor is given by the median of the test statistics, divided by its expectation under the null hypothesis of no association, which is 0.456 in the context of an allelic-effect based analysis . Users hth study . The varjth SNP, and has an approximate χ2 distribution with Nj-1 degrees of freedom under the null hypothesis of consistency where Nj denotes the number of studies for which an allelic effect is reported. An alternative statistic, Qj to variability in the number of studies included in the meta-analysis [To test for consistency of allelic effects across studies at the same SNP, GWAMA calculates two summary statistics of heterogeneity . Cochrananalysis ,18.jth SNP is given byIn the presence of heterogeneity of allelic effects between studies, it is common to perform random-effects meta-analysis in order to correct the deflation in the variance of the fixed-effects estimate . The ranand is used to inflate the variance of the estimated allelic effect in each study. The combined allelic effect across all studies at the SNP is then given by2 distribution with one degree of freedom, and this provides the basis of a test of association of the trait with the jth SNP, allowing for heterogeneity of allelic effects between studies.where p-value; (iv) heterogeneity test statistics Q  and I2; (v) heterogeneity summary, where each study is coded as '+' for increased effect of the reference allele, '-' for decreased effect of the reference allele, '0' for no effect of the reference allele, at a pre-specified significance threshold, and '?' if the study did not report an effect for the SNP. The output from GWAMA can be used with R scripts, supplied with the software, to generate QQ and Manhattan plots to summarise the genome-wide meta-analysis.For each SNP, GWAMA will output a variety of summary information and statistics: (i) reference allele to which effects have been aligned and the corresponding non-reference allele; (ii) meta-analysis allelic effect estimate and standard error ; (iii) meta-analysis association test statistic, and corresponding To demonstrate the utility of GWAMA, we present the results of an example meta-analysis of 5 GWA studies of a simulated quantitative trait with directly typed and imputed genotypes at almost 2.4 million SNPs. Association summary statistics for each individual had previously been corrected for population structure, prior to meta-analysis. Statistical tests of association from the fixed-effects meta-analysis at each SNP were corrected for potential between-study variation on output using genomic control. The analysis was completed in just 3.5 minutes using a dedicated processor with 2.4 Gb memory. The data set used in this example is made available with GWAMA to test individual processor capabilities and potential limitations. To evaluate the memory capacity and program running time, we made additional testing with 20, 50, 100 and 200 genome wide datasets (each containing 2.4 million markers). The GWAMA program peaked with memory usage 4.8 GB, 8.2 GB, 14.6 GB, and 26.2 GB accordingly taking 10 min, 24 min, 53 min, and 1 hour 52 min to run.p-value less than 10-8).Figure There are currently several software packages designed for genome-wide meta-analysis of association test statistics including METAL , MetABELIn the coming months, we expect many more meta-analyses to be undertaken of increasing numbers of GWA studies of a wide range of phenotypes. With the imminent release of data from the 1000 Genomes project , we expeProject name: GWAMAProject home page: http://www.well.ox.ac.uk/GWAMAOperating system: UNIX (source code can be compiled with other platforms), Windows XP and newerProgramming language: C++, R, PERLOther requirements: C++ compiler, optionally R version 2.9.0 or later with PNG support to generate graphics and PERL to run file formatting scriptsLicence: BSDAny restrictions to use by non-academics: noneRM scripted and tested the software. APM provided statistical support and participated in error checking. Both authors wrote and approved the final manuscript.