TP53 gene  and MDM2 promoter , have been independently associated with increased risk of several cancer types. Few studies have analysed the role of these polymorphisms in the development of hepatocellular carcinoma.Single-nucleotide polymorphisms within TP53 codon 72 and MDM2 SNP309 in 61 viral hepatitis-related hepatocellular carcinoma cases and 122 blood samples  from Italian subjects were determined by PCR and restriction fragment length polymorphism (RFLP).Genotype distribution of TP53 codon 72 alleles were not significantly different between cases and controls. A significant increase of MDM2 SNP309 G/G and T/G genotypes were observed among hepatocellular carcinoma cases .Frequencies of MDM2 SNP309 G allele as a susceptibility gene for the development of viral hepatitis-related hepatocellular carcinoma among Italian subjects.These results highlight a significant role of  Hepatocellular carcinoma (HCC) is the most common liver malignancy as well as the third and the fifth cause of cancer death in the world in men and women, respectively . HCC is Etiological factors associated with the development of the disease are well known and include infection with the hepatitis C virus or hepatitis B virus, heavy alcohol intake, prolonged dietary exposure to aflatoxin B1 and primary hemochromatosis -9. As foTP53 gene, resulting in a non-conservative arginine to a proline change at codon 72, is known to be important for growth suppression and apoptotic functions [MDM2 gene has been shown to increase the binding affinity of the transcriptional activator Sp1 resulting in high levels of MDM2 protein, formation of transcriptionally inactive p53-MDM2 complexes and alteration of the p53 pathway [Several studies have analyzed distribution of single nucleotide polymorphism in gene encoding for cell cycle regulatory proteins and susceptibility to malignant diseases. A common polymorphism located in exon 4 of unctions ,13. Addi pathway -16. Thes pathway .TP53 codon 72 and MDM2 SNP309 have been previously investigated in Korean population in which they are associated with the early development of HCC in patients with chronic HBV infection [Polymorphisms of nfection .TP53 codon 72 genotypes and MDM2 SNP309 in a series of 61 hepatocellular carcinoma cases comprising HCV and HBV infected patients, and compared them with healthy controls to possibly identify host genetic factors associated with the increased risk of developing hepatocellular carcinoma.No studies have been conducted in HCC cases of the Italian population. In the present study, we evaluated the distribution of the Liver biopsies from 53 HCV-positive, 7 HBV-positive, one HCV/HBV-positive HCC patients and 122 negative non-liver cancer control patients (blood samples) were obtained with informed consent at the liver unit of the INT "Pascale" in Naples. Each liver biopsy was divided in two sections: the first section was stored in RNA Later at -80°C , the second was subjected to histopathologic examination. Only liver biopsies determined to be hepatocellular carcinoma were included in the study.TP53 and PCR-RFLP of intron 1 of MDM2 [Genomic DNA was extracted according to published procedures . Tissue  of MDM2 ,21.TP53 genotype at codon 72 was carried out by performing an allele specific PCR in which two independent reactions were performed for each sample: one specific for codon CCC (encoding proline) using the p53ProPlus (5'-GCCAGAGGCTGCTCCCCC-3') with p53Minus oligoprimers; the other specific for CGC (encoding arginine), using the p53Plus with p53ArgMinus (5'-CTGGTGCAGGGGCCACGC-3') oligoprimers. PCR amplification reactions were carried out on 100 to 200 ng genomic DNA in 50-μl reaction mixture following amplification procedures described previously [The analysis of MDM2 SNP309 was determined by PCR and restriction fragment length polymorphism (RFLP) analysis of amplified products. A 174 base pairs (bp) amplimer of the MDM2 intron 1 region, containing the MspA1 polymorphic site at nucleotide 309, was amplified with the B-MDM2-309F (5'- GGGAGTTCAGGGTAAAGG-3') and the B-MDM2-309R (5'- GACCAGCTCAAGAGGAAA-3') oligoprimers, designed using the Beacon Designer 2.0 software . PCR reactions were performed in a 50-μl reaction mixture containing 100-200 ng of target DNA, 20 pmol of each primer, 2.5 mM MgCl2, 50 μM of each dNTP and 1.25 U of HotMaster Taq DNA polymerase . DNA was amplified with the following steps: an initial 1-min denaturation at 94°C, followed by 32 cycles of 55°C for 45 s, 68°C for 1 min, 94°C for 30 s and a final annealing at 55°C for 30 s with 5 min elongation at 72°C. Ten-microliter aliquots of MDM2 PCR products were digested with MspA1 restriction enzyme, fractionated by electrophoresis on a 7% polyacrylamide gel in Tris-borate-EDTA running buffer and stained with ethidium bromide for DNA band visualization by UV transillumination.The TP53 PCR amplified products from the 61 cases were further subjected to direct nucleotide sequencing by Primm Srl Laboratories  using the fluorescent dye terminator technology and ABI 3730 DNA sequencers (Applied BioSystems). Nucleotide sequences were edited with Chromas Lite 2.01 (http://www.technelysium.com.au/chromas.html).2 test was used, as appropriate, to compare the proportions of TP53 and MDM2 genotypes between cases of HCC and healthy control population. An unpaired Student's t-test was used to evaluate differences between the mean age of cases and control group. All analyses were performed with Epi Info 6 Statistical Analysis System Software .The observed and expected genotype frequencies among the study groups were analysed using the Hardy-Weinberg equilibrium theory. A Fisher's exact test or χp-values were less than 0.05.Differences were considered to be statistically significant when TP53 codon 72 genotypes has been estimated using an allele specific PCR that specifically detects either the TP53 proline or arginine 72- encoding codons. A slight higher frequency of arginine allele was observed in HCC cases compared to the controls, however, the difference did not reach statistical significance using Yates corrected χ2 test (P = 0.696). The distribution of polymorphic alleles in cases and controls are summarized in Table TP53 gene polymorphism at codon 72 among the 61 HCC cases were 4.9% (n = 3), 32.8% (n = 20) and 62.3% (n = 38) proline homozygous, heterozygous and arginine homozygous, respectively, and the corresponding figures among controls were 7.4% (n = 9), 34.4% (n = 42), and 58.2% (n = 71), respectively. The genotype frequencies of TP53 codon 72 of were found in Hardy-Weinberg equilibrium both in HCC cases and healthy controls .In this study were included 61 cases of hepatocellular carcinoma  and 122 cancer-free control subjects . Table MDM2 SNP309 genotypes were analyzed using a PCR-RFLP based assay , 52.5% (n = 32) and 26.2% (n = 16), and among controls were 45.1% (n = 55), 39.3% (n = 48) and 15.6% (n = 19), respectively. The MDM2 SNP309 genotype frequencies were found in Hardy-Weinberg equilibrium among cases and controls group . The frequencies of MDM2 SNP309 G/G and G/T genotypes were significantly more prevalent in HCC cases (52.5%) compared with controls (36.9%), p = 0.006. When the homozygous MDM2 SNP309 T/T genotype was used as the reference group, the MDM2 SNP309 G/G and T/G genotypes were associated with a significantly increased risk for HCC .The y Figure . The freThe combined effect of the TP53 codon72 and the MDM2 SNP309 polymorphisms was also analysed. However, no additional cancer risk effect associated with TP53 codon 72 polymorphism was identified when the risk of combined genotypes was analyzed.TP53 codon 72 and MDM2 SNP309 polymorphisms and risk of or survival from several types of cancer [Several epidemiological studies have evaluated the association of f cancer -24, but TP53 codon 72 and MDM2 nucleotide 309 polymorphisms were investigated in a series of 61 cases of hepatocellular carcinoma, mainly associated with HCV infection, and in 122 population-matched controls in order to verify the impact of these gene variants on the risk of tumour development. The TP53 codon 72 and MDM2 SNP309 genotypes were distributed in accordance with the Hardy-Weinberg equilibrium in controls and their frequencies were in broad agreement with those observed in previously published studies of Caucasian subjects [TP53 codon 72 polymorphisms between cases and controls and no increase in the frequency of the arginine or proline alleles among HCC cases. This observation is in agreement with previous studies reporting no association between codon 72 genotypes and disease severity or liver cancer in a group of 340 HCV-infected Italian subjects at different stages of disease, including 84 HCC patients, and in a group of 97 Spanish HCC patients [Pro/Pro genotype [TP53 codon 72 Pro allele and HCC was apparent. Indeed, in patients with chronic liver disease there were synergistic effects between the Pro allele and HCC family history in first-degree relatives which conferred a risk of 7.60 (95% CI = 2.28-25.31) for HCC development compared with subjects without the Pro allele and without chronic liver disease [TP53 codon 72 polymorphism in the HCC pathogenesis in different populations.In the current study the subjects -27. The patients ,29. Convgenotype . A case- disease . These oMDM2 SNP309 T/G heterozygous (52.5%) and G/G homozygous (26.2%) genotypes were significantly higher among HCC cases compared to healthy controls . The homozygous MDM2 SNP309 T/T genotype in HCC, on the other hand, was lower (21.3%) than that observed in controls (45.1%). The possible role of MDM2 SNP309 polymorphism in HCC development has been analyzed in few other geographical regions. In Eastern Asia MDM2 SNP309 G allele has been associated with higher risk of HCC in Japanese patients with chronic hepatitis C [et al. (2011) performed a metanalysis on these studies, including a total of 738 cases and 1062 controls, to evaluate the reliability of the associations of MDM2 SNP309 G allele with HCC by using false-positive report probability analysis and the Venice guidelines on genetic epidemiology [In this study the frequencies of atitis C , in Koreatitis C , and in atitis C . Similaratitis C  and in Tatitis C . Jin et emiology . The pooemiology .Overexpression of the MDM2 protein has an oncogenic effect through the reduction of p53 levels with the consequent attenuation of the p53 DNA damage response that allows increased cell proliferation and inhibition of apoptosis, providing advantageous signals for tumor cell survival -40. The TP53 and MDM2 polymorphisms in the control group, it may be excluded that age and sex limitations have introduced significant bias in the study.This study has limitations, including the modest sample size and the fact that cases were not well matched for age and sex with the control subjects. Although Italian blood donors may be good representatives of the general population, they are younger than HCC patients due to age limit and health requirements for blood donation. However, being fulfilled the Hardy Weinberg equilibrium for the allelic frequencies of both TP53 polymorphism at codon 72 seems not to be a potential risk factor for development of HCC in Italian patients.In conclusion, these results provide the first evidence that the MDM2 SNP309 polymorphism represents a risk factor for viral hepatitis-related HCC in the Italian population. Conversely, the MDM2: mouse double minute-2; HBV: hepatitis B virus; HCV: hepatitis C virus.HCC: Hepatocellular carcinoma; The authors declare that they have no competing interests.MLT and FMB were responsible for the overall planning and coordination of the study. FI was involved in patients enrollment and specimen collection. LB contributed to the data analysis. SL and GB carried out the histopathology evaluation of the cases. VDV was responsible for specimen processing, DNA analysis and with MLT compiled and finalized the manuscript. All authors read and approved the final manuscript.


To analyze the association between −1082A/G polymorphism in interleukin-10 (IL-10) gene and ischemic stroke (IS) risk by meta-analysis.We carried out a systematic electronic search in PubMed, BIOSIS Previews, Science Direct, Chinese National Knowledge Infrastructure, Chinese Biomedical Database, Weipu database and WANGFANG Database. Pooled odds ratios (ORs) with 95% confidence intervals (95%CIs) were calculated to assess the strength of the association.P = 0.0008; AA+AG vs. GG OR 1.54, 95% CI 1.18–2.01, P = 0.001).7 studies were included. There was no significant association between IL-10 −1082A/G polymorphism and IS risk under all genetic models in overall estimates . Similarly, no associations were found in subgroup analysis based on ethnicity and source of controls. However, removing the study deviating from Hardy–Weinberg equilibrium (HWE) produced statistically significant associations for overall estimates under recessive model and among Asians in all genetic models . Even after Bonferroni correction, the associations were observed still significantly in Asians under the two models (AA vs. GG OR1.91, 95% CI 1.31–2.80, This meta-analysis indicates that IL10 −1082 A/G polymorphism is associated with IS susceptibility in Asians and the −1082 A allele may increase risk of IS in Asians. Considering the sample size is small and between-study heterogeneity is remarkable, more studies with subtle design are warranted in future. Ischemic stroke (IS) is a major cause of adult disability and death in the worldIL-10(Gene ID: 3586) is a multifunctional cytokine with anti-inflammatory properties, which has been showed involving in the inflammatory process of ISEmerging studies have reported the associations between −1082G/A polymorphism in IL-10 gene and IS riskhttp://sinomed.imicams.ac.cn/index.jsp), Weipu database (http://cstj.cqvip.com/), Chinese National Knowledge Infrastructure (http://dlib3.cnki.net/kns50/),and WANFANG Database (http://g.wanfangdata.com.cn/) up to 10th October 2013 to identify relevant studies, using the following key words: (“interleukin-10” OR “interleukin 10” OR “IL-10” OR “IL10”) and  and . In addition, hand searching of the references in selected literatures and the abstracts presented at relevant conferences were performed for other potential related studies. Languages were limited to English and Chinese.This meta-analysis conformed to the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) criteria Studies meeting the following criteria were included:(1) the study should evaluate the relationship between IL-10 gene −1082A/G polymorphism and IS risk;(2) the study had to be a case-control design; (3) genotype distributions in both cases and controls were available for calculating an odds ratio (OR) with 95% confidence interval (CI); (4)Computed tomographic(CT) or magnetic resonance imaging(MRI) were used to assess the diagnosis of IS. The following were exclusion criteria: (1) reviews, abstracts or animal studies; (2)studies were not relevant to IL-10 gene −1082A/G polymorphism or IS; (3) studies did not report genotype frequencies;(4) the design were based on sibling pairs or family. If studies were repeated or overlapped publications, the most complete one was included. If studies did not report detailed data, we would get in touch with authors to obtain the relevant information.Two investigators (Jin and Li) reviewed and extracted data independently in accordance with the inclusion criteria. The results were compared, and if any disagreement appeared, a third investigator (Peng) was invited to evaluate such studies, then the discrepancy was resolved by discussion. The following information were extracted: the name of first author, year of publication, country (ethnicity), diagnostic criteria of IS, study design, sample size, allele numbers and genotype distributions in cases and controls.The quality of included studies were assessed by 2 investigators (Jin and Li) independently on the basis of Newcastle-Ottawa Scale (NOS)P<0.05 was considered as statistically significant. The Q-test and I2-statistics were employed for evaluating the between-study heterogeneity, which was considered as significant when QP≤0.10 or I2>50%QP≤0.10 or I2>50%) or absence (QP>0.10 or I2≤50%) of heterogeneity, respectively. Furthermore,to explore the sources of heterogeneity, we conducted subgroup analysis based on ethnicity, source of controls,respectively. Moreover, Bonferroni method, controlling for false positive error rate, was utilized to adjust for multiple comparisons. As we performed multiple comparisons in this meta-analysis for 25 times, the P value which was less than 0.05/25 (0.002) indicated statistical significance after Bonferroni correction. To validate the reliability of the results, sensitivity analysis was performed though omitting one case–control study each time, as well as limiting this meta-analysis to studies which were conformed to HWE. HWE of genotype distribution in the controls of included studies was conducted using an online program (http://ihg2.helmholtz-muenchen.de/cgi-bin/hw/hwa1.pl), and P<0.05 was considered significantly deviating from HWE. Publication bias was evaluated by visual inspection of symmetry of Begg's funnel plot and assessment of Egger's testP<0.05 was regarded as representative of statistical significance). All statistical analyses were performed using software RevMan 5.1 and STATA 11.0.Pooled ORs with corresponding 95% CIs were calculated to evaluate the strength of relationship between IL 10 gene −1082 A/G polymorphism and IS risk under the following five genetic models: the allele model (A vs. G), the homozygote model (AA vs. GG), the heterozygote model (AG vs. GG), the dominant model (AA+AG vs. GG), and the recessive model (AA vs. AG+GG). Z-test was used to assess the significance of the pooled OR, in which The detailed characteristics of the included studies are listed in All included studies were of high quality as the NOS score of each one was higher than 5 points and the genotype distributions in all controls were consistent with HWE except 1 studyAs shown in Notably, there was no GG genotype in either case or control group of both the Lin's study and Zhang's studyTo explore the sources of heterogeneity, we performed further subgroup analyses by ethnicity and source of controls respectively. Similarly, there were no significant associations in the subgroup analyses, and significant heterogeneity in most of the comparison models still existed. P = 0.02; AA vs. GG: OR 1.91, 95% CI 1.31–2.80,P = 0.0008; AG vs. GG: OR1.44, 95% CI 1.09–1.91,P = 0.01 AA+AG vs. GG: OR1.54, 95% CI 1.18–2.01,P = 0.001,AA VS. AG+GG: OR 1.79, 95% CI 1.07–3.00, P = 0.03, whereas the corresponding pooled ORs of total available studies which meant overall analysis for all ethnicities were not materially altered (data not shown), except in recessive model. However, after Bonferronic correction for multiple testing, the associations revealed still significantly in Asian subgroup under homozygote and dominant models . The results indicated that the homozygote AA and A allele carriers (AA+AG) had nearly a 91% and 54% increased risk of IS respectively, when compared with the homozygote GG in Asians. In particular, removing the study which was deviated from HWE eliminated the heterogeneity in subgroup analysis of Asians in homozygote, heterozygote, and dominant models, but not in the allele and recessive models and the pooling analysis   .It is now accepted that genetics and environmental factors contribute to IS susceptibility and outcome. Similarly, the inflammation reaction is also relevant to IS. IL-10 is a potent anti-inflammatory cytokine with multiple functions taking part in inflammation reaction as well as the development of IS. Recently, the associations between IL-10 gene −1082 A/G polymorphism and the risk of IS have been intensively investigated, however, the results are inconsistent. Therefore, we designed this meta-analysis to draw a more precise conclusion for the association between IL-10 −1082 A/G polymorphism and IS risk.In this meta-analysis, no association of the IL-10 −1082A/G polymorphism with IS risk was found under all comparisons, and in subgroup analysis by ethnicity or source of controls. However, for Asians, after excluding the study deviating from HWESignificant between-study heterogeneity displayed among all comparison models. Considering that the diversity in design, difference of ethnicity, sample sizes, and measurement errors may contribute to common sources of heterogeneity As far as we know, this is the first comprehensive meta-analysis exploring the association between IL-10 −1082 A/G polymorphism and IS risk up to now, which involved Caucasian, Indian and Chinese populations. In addition, more studies were included in our study than a recently published meta-analysis concerning −1082A/G polymorphism and IS risk only in South AsiansDespite of the advantages mentioned above, the current study has some inevitable limitations that should be acknowledged. First, there was significant heterogeneity among included studies. Even though we used the random-effects model to calculate pool ORs, the precision of outcome would be affected. Second, owing to limiting detailed information such as lacking of subtyping of the ischemic strokes in individual study, we failed to perform further subgroup analysis to adjust these possible confounders. Third, the sample size in individual included studies was respectively small, especially in homozygote, heterozygote and dominant models. Forth, only English and Chinese language studies were included in this meta-analysis which might have led to bias. Despite of conducting an exhaustive search for eligible studies, some relevant studies so called “grey literatures” might be still missed. Fifth, similar to a case–control study, meta-analysis was a retrospective study, which might lead to recall bias. Last but not the least, this study only explored one variation in IL-10 gene, which ignored such gene–gene and gene-environmental interactions.In conclusion,this meta-analysis indicates that IL- 10 gene −1082 A/G polymorphism is associated with IS susceptibility in Asians and the −1082 A allele may increase risk of IS in Asian populations. However, considering the limitations mentioned above, more well designed studies with adequately sized populations are needed in future.Checklist S1PRISMA 2009 Checklist.(DOC)Click here for additional data file.


Existing literature indicates that ADRB2 gene is associated with health and longevity, but none of previous studies investigated associations of carrying the ADRB2 minor alleles and interactions between ADRB2 genotypes and social/behavioral factors(GxE) with health outcomes at advanced ages. This study intends to fill in this research gap.We conducted an exploratory analysis, using longitudinal survey phenotype/genotype data from 877 oldest-old aged 90+. To estimate association of GxE interactions with health outcome, adjusted for the potential correlation between genotypes and social/behavioral factors and various other potentially confounding factors, we develop and test an innovative three-step procedure which combines logistic regression and structural equation methods.rs1042718 minor allele is significantly and positively associated with good cognitive function; interaction between regular exercise and carrying rs1042718 or rs1042719 minor allele is significantly and positively associated with self-reported good health; and interaction between social-leisure activities and carrying rs1042719 minor allele is significantly and positively associated with self-reported good health. Carrying rs1042718 or rs1042719 minor alleles is significantly and negatively associated with negative emotion, but the ADRB2 SNPs are not significantly associated with cognitive function and self-reported health. Our structural equation analysis found that, adjusted for the confounding effects of correlation of the ADRB2 SNPs with negative emotion, interaction between negative emotion and carrying rs1042718 or rs1042719 minor allele is significantly and negatively associated with cognitive function. The positive association of regular exercise and social-leisure activities with cognitive function and self-reported health, and negative association of negative emotion with cognitive function, were much stronger among carriers of rs1042718 or rs1042719 alleles, compared to the non-carriers.Interaction between regular exercise and carrying rs1042718 or rs1042719 minor alleles with negative emotion, at advanced ages in China. Our findings are exploratory rather than causal conclusions. This study implies that near-future health promotion programs considering individuals’ genetic profiles, with appropriate protection of privacy/confidentiality, would yield increased benefits and reduced costs to the programs and their participants.The results indicate significant positive associations of interactions between social/behavioral factors and the ADRB2 genotypes with health outcomes of cognitive function and self-reported health, and negative associations of carrying  ADRB2 gene appears to be associated with a broad range of aging-associated phenotypes, including cancers at different sites, myocardial infarction, intermittent claudication, and longevity  and cognitive function. To our knowledge, this is the first such attempt based on our innovative three-step procedure which combines the logistic regression and structural equations methods  or 5% (p < 0.05) may not need to be corrected, because the two outcome indicators and multiple explanatory factors in our case are complementary and not independent of each other. For example, both SNPs of rs1042718 and rs1042719 are highly correlated and in strong linkage disequilibrium (70%) [The biostatistics literature states that measuring multiple health outcomes and explanatory factors can lead to multiple comparisons and p values may need to be adjusted by the Bonferroni method or other similar method; however, if the null hypotheses are not independent and the multiple comparisons are complementary, then the p values may not need to be corrected ,60. Our um (70%) . Obviousrs1042718 minor allele is significantly associated with cognitive function score (p <0.05); GxE interactions between regular exercise and carrying rs1042718 or rs1042719 minor allele are significantly associated with self-reported good health (p <0.05 or p <0.01); and GxE interactions between social-/leisure activities and carrying rs1042719 minor allele are significantly associated with self-reported good health (p <0.05) . We also found that carrying the rs1042718 minor allele or rs1042719 minor allele is significantly associated with negative emotion (p <0.05). The rGE correlation between carrying the rs1042718 or rs1042719 minor allele and negative emotion is indirectly associated with cognitive function, but the estimates are not statistically significant  and cognitive function or self-reported health are much stronger among carriers of the ADRB2 minor alleles and their interactions with social/behavioral factors may affect health outcome at old ages. Further studies need to extend the analysis to explore the effects of GxG interactions between ADRB2 genotypes and the other genotypes associated with health and longevity at old ages (e.g. APOE4 and FOXO genotypes), and to cover all elderly age groups  to fully understand the process of healthy aging in a life course perspective. We also hope that other studies on healthy longevity involving similar or substantially larger samples of the oldest-old aged 90+ will replicate and validate our exploratory findings.As detailed in the discussion section, while we are satisfied with the interesting and unique findings reported in this article, our study also has important limitations and unanswered questions; our findings are typically exploratory rather than any kind of causal conclusions. Clearly, additional in-depth epigenetics studies with more sophisticated psychological indicators are needed to develop a deeper understanding of the biological mechanisms and causalities of how and why carrying the aThese 22 provinces are: Liaoning, Jilin, Heilongjiang, Hebei, Beijing, Tianjing, Shanxi, Shaanxi, Shanghai, Jiangsu, Zhejiang, Anhui, Fujian, Jiangxi, Shangdong, Henan, Hubei, Hunan, Guangdong, Guangxi, Sichuan, Chongqing.bIn a dominant model, any genotype that contains 1 or 2 copies of the minor allele is coded as 1; and otherwise the genotype that does not contain copy of the minor allele is coded as 0 . For the recessive model, the homozygous mm genotype is coded as 1 and 0 otherwise . In an additive model, genotype MM is coded as 0, Mm, 1, mm, 2.cNote that Zhao et al. [o et al.  basicallo et al. . HoweverdMMSE is used internationally in healthy aging investigations and works well in studies that are not specialized psychological research.eWe also tried the continuous MMSE scores, the four categories  of MMSE scores and the dichotomized MMSE scores. The results of the continuous and the alternative classifications of MMSE scores are generally consistent with the estimates using the three categories of the MMSE scores, while the significance level reduced moderately in the continuous model and slightly in the four categories and dichotomized models, as compared to the three-category model we adopted.fAbout 78% of the Chinese oldest-old aged 90+ had no schooling.gParticipation in the social-leisure activities depends on senior adults’ interests and it is also subject to time limit. Thus, it is not necessarily the case that the larger number of social-leisure activities an elder participated in, the more active he/she is, and it may not make sense to use a continuous variable to measure the social-leisure activities. After trying different combinations, we found that dichotomization of the social-leisure activities score as ≥2 vs. <2 is the best choice.hNote that in the structural equation model (SEM) and M-Plus software used for this study, one needs to create a new variable for the interaction term between the genetic variant and social/behavioral factor [l factor . We use iWe conducted the statistical analysis models by pooling males and females together, while controlling for gender as a covariate. We also tried the logistic regression analysis for males and females separately and the results are basically consistent with our presented two-genders combined models; the statistical significance levels were reduced substantially due to sample size reduction.The authors declare that they have no competing interests.YZ designed the research; supervised data collection and data analysis, and wrote the paper; XLT designed the genetic research and supervised the lab work; LGC, QSF, HSC and KS analyzed data; LJX, LGC, FYZ, HSC, SK performed data collection; LZ and HQC performed lab tests; ZY, JG and WT contributed to sample storage; QHT, SGG, ZY, JG, WT, XLT and ERH suggested revisions of the paper. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2318/13/91/prepub


Areca nut chewing has been reported to be associated with obesity, metabolic syndrome, hypertension, and cardiovascular mortality in previous studies. The aim of this study was to examine whether chewing areca nut increases the risk of coronary artery disease (CAD) in Taiwanese men.This study is a hospital-based case-control study. The case patients were male patients diagnosed in Taiwan between 1996 and 2009 as having a positive Treadmill exercise test or a positive finding on the Thallium-201 single-photon emission computed tomography myocardial perfusion imaging. The case patients were further evaluated by coronary angiography to confirm their CAD. Obstructive CAD was defined as a ≥ 50% decrease in the luminal diameter of one major coronary artery. The patients who did not fulfill the above criteria of obstructive CAD were excluded.The potential controls were males who visited the same hospital for health check-ups and had a normal electrocardiogram but no history of ischemic heart disease or CAD during the time period that the case patients were diagnosed. The eligible controls were randomly selected and frequency-matched with the case patients based on age. Multiple logistic regression analyses were used to estimate the odds ratio of areca nut chewing and the risk of obstructive CAD.A total of 293 obstructive CAD patients and 720 healthy controls, all men, were analyzed. Subjects who chewed areca nut had a 3.5-fold increased risk (95% CI = 2.0-6.2) of having obstructive CAD than those without, after adjusting for other significant covariates. The dose-response relationship of chewing areca nut and the risk of obstructive CAD was also noted. After adjusting for other covariates, the 2-way additive interactions for obstructive CAD risk were also significant between areca nut use and cigarette smoking, hypertension and dyslipidemia.Long-term areca nut chewing was an independent risk factor of obstructive CAD in Taiwanese men. Interactive effects between chewing areca nut and cigarette smoking, hypertension, and dyslipidemia were also observed for CAD risk. Further exploration of their underlying mechanisms is necessary. Ischemic heart disease, including coronary artery disease (CAD), is the leading cause of death worldwide . CAD patAreca nut chewing is the fourth most popular substance abuse habit in the world . PreviouThis study is a hospital-based case-control study. The case patients were male patients diagnosed as having a positive treadmill exercise test or a positive finding on the Thallium-201 (Tl-201) single-photon emission computed tomography (SPECT) myocardial perfusion imaging (MPI) at the Kaohsiung Medical University Hospital (KMUH), a medical center in the southern Taiwan, between 1996 and 2009. The positive Treadmill exercise test was defined as an ST segment elevation or depression ≥ 1 mm in three consecutive beats in ≥ 2 consecutive leads. A positive finding on the Tl-201 SPECT MPI was defined as reversible defects in at least one vascular territory. Case patients were also performed a coronary angiography (CAG) using the Judkins technique. Then, the angiograms were analyzed by two experienced cardiologists who were blinded to the results of the questionnaire. The severity of stenosis was assessed by the quantitative coronary angiography  of the major coronary arteries and their major branches. Obstructive CAD was defined as a ≥ 50% decrease in the luminal diameter of one major coronary artery ,13. If aThe potential controls were males who visited the same hospital for health check-ups during the same time period as the case patients. The potential controls had a normal electrocardiogram record during their examination and did not have any history of ischemic heart disease or CAD. The eligible controls were randomly selected and frequency-matched with the case patients based on age . In total, 293 case patients and 720 healthy controls were included in this study. This study was approved by the ethics committee of KMUH, and all study subjects provided written informed consent.A standardized questionnaire was used by a well trained research assistant to interview study subjects. The questionnaire collected detailed information about the subjects' demographic characteristics and history of using three major substances . The detailed information has been described elsewhere ,15. In bTwo kinds of chewing areca nut commonly used in Taiwan were also included on the questionnaire: Lao-hwa and betel leaf. The composite of the Lao-hwa regimen includes areca nut, betel palm inflorescence, and slaked lime, whereas the composite of betel leaf regimen includes areca nut, betel leaf, and slaked lime .p = 0.0028) [The items on the questionnaire regarding the use of these three major substances were validated in our previous study ,15. In b 0.0028) .We quantified the cumulative amount of substance use by "drink-years" for alcohol consumption and "pack-years" for cigarette smoking and areca nut chewing. The number of "drink-years" and "pack-years" were calculated by multiplying the amount of the substances consumed per day, such as 15.75 g-alcohol drinks , 20-cigarette packs or 10-areca nut packs, by the years of substance use.Personal disease histories including hypertension (HTN), diabetes mellitus (DM), dyslipidemia, stroke, CAD, and arrhythmia were also recorded. HTN was defined as having a history of HTN or anti-HTN drug use. DM was defined as having a history of DM, using an oral glucose-lowering agent or insulin, or having a fasting blood sugar greater than 126 mg/dL. Dyslipidemia was defined as having a history of dyslipidemia, using a lipid-lowering agent or having an abnormal high level of plasma cholesterol (>200 mg/dL) during health exams.Demographic characteristics, histories of substance use and prevalence of disease were compared between healthy controls and case patients (obstructive CAD patients) by Chi-Square test or Fisher's exact test when appropriate. Because our main study aim was to investigate whether chewing areca nut is a risk factor for CAD, we used unconditional logistic regression to analyze the effect of different levels of areca nut use on CAD risk, after adjusting for other covariates that were statistically significant in the crude analyses. These covariates included body mass index (BMI), educational level, age, cigarette smoking, alcohol consumption, DM, HTN, and dyslipidemia.p value less than 0.05. All statistical analyses were carried out by using SAS software .To assess the interactive effect of areca nut use and other main risk factors  for CAD among subjects, Rothman's synergism index (SI) was used to assess the empirical deviation from the additive interaction relation . A compuThe percentages of CAD patients in the ≤ 40, 41-60, and > 60 year old age groups were 3.4% (10/293), 37.2% (109/293) and 59.4% (174/293), respectively. These percentages were similar to those of the healthy controls. Compared to healthy controls, the obstructive CAD patient groups had a significantly higher percentage of DM, HTN, dyslipidemia, cigarette smoking, and areca nut chewing. In addition, obstructive CAD patients were less educated, and more likely to be obese than the healthy controls  of having obstructive CAD  and 8.0-fold higher risks (95% CI = 3.6-17.9) of CAD for 1-20 pack-years and > 20 pack-years of areca nut chewing, respectively  = 1.19, 95% CI: 1.05, 1.35) and cerebrovascular disease mortality , compared with those who never chewed areca nut . In the To minimize outcome misclassification, the present study used CAG to confirm the diagnosis of obstructive CAD. In addition, the exposure of interest (areca nut chewing) in this study was validated in our previous research . The preWe also found a dose-dependent relationship between areca nut chewing and CAD risk, as a higher amount of areca nut chewing was associated with a higher risk of obstructive CAD. The additive interactions in the risk of obstructive CAD between "areca nut chewing and cigarette smoking," "areca nut chewing and HTN" and "areca nut chewing and dyslipidemia" were also observed. CAD, which is similar to other atherosclerotic diseases, has been previously associated with risk factors such as DM, HTN, dyslipidemia, age and obesity . Areca nThe present study found that the Lao-hwa regimen was more potent than the betel leaf regimen by comparing the ORs for their associations with obstructive CAD. Betel leaf may have some beneficial effects for cardiovascular disease through its vasorelaxation, antioxidant effects and inhibition of platelet aggregation -27, whicin-vitro studies [There are several possible mechanisms to explain the link between areca nut chewing, CAD and atherosclerosis. Atherosclerosis, especially CAD, was related to chronic inflammation in previous studies ,28. Arec studies ,30. Hydr studies , increas studies  and redu studies ,34. One  studies . Subsequ studies . The exp studies . The ext studies . BecauseThere were several limitations in the study. The healthy controls were drawn from healthy volunteers attending general health check-ups; thus, this group may not be representative of the general population. The detailed lifestyle information about vegetable consumption and exercise status was not routinely collected in our questionnaire, which might influence the risk of CAD studied here.In conclusion, we have shown that long-term areca nut chewing is an independent risk factor of CAD in Taiwanese men, especially among subjects who smoke cigarettes. A dose-response effect of areca nut chewing in relation to the risk of CAD was also found in this study. There is an additive effect of areca nut chewing and cigarette smoking, HTN, and dyslipidemia on the risk of CAD. Further studies are needed to clarify the mechanisms of areca nut chewing leading to CAD.BMI: Body mass index; CAD: Coronary artery disease; CAG: Coronary angiography; CI: Confidence intervals; DM: Diabetes mellitus; HTN: Hypertension; IL: Interleukin; LDL: Low-density lipoprotein; MPI: Myocardial perfusion imaging; OR: Odds ratio; SI: Synergism index; SPECT: Single-photon emission computed tomography; Tl-201: Thallium-201.The authors declare that they have no competing interests.TWC, WMT and LWT designed the study; WGJ, LKT, LYH, YHW, CCS, CYT, LTH, SHM, HPC, CKH, DTH, KYC and SSH conducted the research; LCH, WMT and TWC analyzed data; and TWC and WMT wrote the paper. TWC, WMT and LWT had the primary responsibility for the final content. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2458/12/162/prepubTable 1. Odds ratio for coronary artery disease associated with areca nuts chewing in male smoking subjects.Click here for fileTable 2. Odds ratio for obstructive coronary artery disease associated with areca nut use.Click here for fileTable 3. The additive interaction between conventional risk factors and areca nuts use for coronary artery disease.Click here for file


Objectives: A number of studies conducted to assess the association between Clara cell 10-kDa protein (CC10) +38A/G polymorphism and susceptibility to asthma have yielded inconsistent and inconclusive results. In the present study, the possible association was assessed by a meta-analysis.Methods: Relevant articles were identified for the period ranging from Jan 1998 up to March 2013. Pooled odds ratios (OR) with 95% confidence intervals (CI) were appropriately derived from fixed effects or random-effects models.Results: Ten case-control studies with a total of 1529 asthma cases and 2399 controls were included in this meta-analysis. The association between CC10 +38A/G polymorphism and asthma risk was determined in dominant model, recessive model, additive model, and codominant model. In dominant model, CC10 +38A/G polymorphism seemed to be associated with elevated asthma risk . Subgroup analyses by ethnicity also found significant associations between this polymorphism and asthma risk in Asians and Caucasians. Results from other genetic models further identified this possible association.Conclusion: This meta-analysis suggests that CC10 +38A/G polymorphism confers asthma risk. Although environmental factors are important determinants of asthma, numerous studies have revealed that asthma has a strong genetic component.2 The CC10 gene is located on chromosome 11q12-13, a region that has been associated with asthma and atopy in several genome-wide linkage studies.3,4 A single nucleotide polymorphism, the +38A/G polymorphism (rs3741240), has been identified in this gene.5 Some papers have reported that this polymorphism was associated with asthma risk, whereas others found no such association.5-14 It was possible that most of these studies only included a modest sample size, and thus each of them might not achieve a reliable conclusion. Meta-analysis is a useful method for investigating associations between genetic factors and diseases, because a quantitative approach is used to combine the results from different studies on the same topic, thereby providing more reliable conclusions.15 Here, we use meta-analysis to clarify the association of CC10 +38A/G polymorphism with asthma. To our knowledge, this is the first genetic meta-analysis of the association between CC10 +38A/G polymorphism and the risk of asthma.Clara cell 10-kDa protein  is a 10-kDa protein produced by non-ciliated Clara cells and is one of the most abundant proteins in the fluids lining the airways.Search for publications: We performed a literature search using the MEDLINE and China National Knowledge Infrastructure (CNKI) databases to identify articles that examined associations between CC10 +38A/G polymorphism and asthma. The following key words and subject terms were searched: Clara cell 10-kDa protein, CC10, CC16, SCGB1A1, uteroglobin, asthma, genetic, and polymorphism. Reference lists of articles retained for review were inspected for relevant publications. No publication date or language restrictions were imposed.Inclusion and exclusion criteria: The following inclusion criteria were used: (i) the study should have evaluated the association between the +38A/G polymorphism and asthma risk; (ii) the study should have had a case-control design; (iii) sufficient data should have been provided in order to calculate odds ratios (OR) and 95% confidence intervals (CI). Studies were excluded if any of the following conditions applied: (i) only abstracts or reviews were available, without sufficient data; (ii) genotype frequencies were not reported; (iii) studies were repeated or publications overlapped.Data extraction: The following information was extracted from each study: author, year of publication, ethnicity of the study population, numbers of cases and controls, and CC10 +38A/G genotype numbers.Statistical analysis: Meta-analyses were performed using dominant model (AA + AG vs GG). Heterogeneity among studies was assessed using the chi-square based Q and I2 statistics. Heterogeneity was considered to be significant for I2 > 50%. Fixed-effects or random-effects models were used to pool the data according to heterogeneity. Other comparative genetic models were also used to assess the association between the polymorphism and the risk of asthma . For the subgroup analysis by ethnicity, the study populations were stratified into two groups: Asian and Caucasian. Galbraith plot was used to spot the outliers which were the sources of heterogeneity. Funnel plot and Egger’s test were used to detect publication bias.16 Analyses were performed using Revman 5.1 and STATA 11.0 software.Characteristics of studies: A total of 10 relevant articles on CC10 +38A/G polymorphism in asthma met the study inclusion criteria , and the Z-test value for overall effect was 3.47 (P = 0.0005) for the AA + AG vs GG. Subgroup analysis by ethnicity was performed. For ethnicity, the populations were stratified into two groups: Asian (908 cases and 784 controls), Caucasian (621 cases and 1615 controls). There were significant associations with asthma risk in these populations: Asian  and Caucasian . Publication bias was assessed graphically by using funnel plots and assessed statistically by using Egger’s test. The shape of the Begg’s funnel plots appeared symmetrical for the AA + AG vs GG model (P = 0.99). Summary results from the other comparisons are presented in GG model . Egger’sHeterogeneity analysis: The between-study heterogeneity was obvious in dominant model (I2 = 65%). Galbraith plots spotted Sharma’s study and Chen’s study as the outliers and the major source of the heterogeneity (I2 = 43%). Besides, a significant association was also observed after excluding Sharma’s study and Chen’s study .ogeneity . The het17,18 Deficiency of CC10 aggravates pulmonary allergic inflammation through augmentation of the TH2 response, and reconstitution of CC10 in CC10-deficient mice is able to reverse the altered phenotypes.19 These results suggested that CC10 played an important role in the development of asthma. Although asthma can have many causes, genetic factors are considered strong determinants, and therefore, researchers have searched for the genes responsible. Several studies have investigated the role of CC10 +38A/G polymorphism in asthma susceptibility both in Asians and Caucasians, with contrasting results. Meta-analysis has been recognized as an important tool to more precisely define the effect of selected genetic polymorphisms on risk of disease. Therefore, we performed the meta-analysis specifically to assess this association.CC10 with primary expression in the uterus and non-ciliated bronchiolar cells, has an anti-inflammatory effect on the urogenital and respiratory tracts.CC10 +38A/G polymorphism and asthma risk . In addition, subgroup analysis by ethnicity showed that this polymorphism was significantly associated with increased asthma risk in both the Caucasians  and Asians . Moreover, other genetic models also found positive results. Thus, meta-analysis of available data suggests CC10 +38A allele contributes to increased asthma risk.The present meta-analysis found a significant association between the I2 = 65%). Galbraith plot was used to spot the outliers as the possible major sources of heterogeneity. Two studies were spotted as the outliers. After excluding those two studies, the between-study heterogeneity decreased and there was no obvious heterogeneity among the left 8 studies (I2 = 43%), which suggested the heterogeneity might come from those two studies. Meta-analysis of the left 8 studies also showed CC10 +38A/G polymorphism was associated with increased risk of asthma .The importance of heterogeneity should be mentioned, as it is an important issue when interpreting the results of meta-analyses. In this present meta-analysis, we found obvious heterogeneity (Some possible limitations in this meta-analysis should be acknowledged. First, only published studies that were included in the selected electronic databases were identified; it is possible that some relevant published or unpublished studies that supported the null hypothesis may have been missed, which may have biased the results, and this may not have been detected by the statistical tests that were performed. Second, the effect of gene-gene and gene-environment interactions was not addressed in this meta-analysis status, because of limited available data. Third, all the data were used without adjustment by detailed individual information such as age, sex, and lifestyle in our meta-analysis. Finally, the number of included studies was moderate.CC10 +38A/G polymorphism confers asthma risk. Further studies can assess the possible gene-environmental and gene-gene interactions in the association between this polymorphism and asthma risk.To sum up, this meta-analysis suggests that Guangri Zhao, Xiaodan Lin, Ming Zhou and Jian Zhao: All the authors had substantial contributions to conception and design, acquisition of data, analysis and interpretation of data, drafting the manuscript and final approval of the version to be published.


It is well known that the effects of exercise training gives inter individual differences which might be due to genetic diversity. This study aims to explore the probable relation between angiotensin converting enzyme (ACE) alleles and physical fitness parameters in elite athletes.2 max, muscular endurance, flexibility and power. DNA was extracted from blood using standard phenol-chloroform method. Genotyping was done using PCR specific for ACE (I/D) polymorphism, followed by agarose gel electrophoresis method. Variation of the parameters among different genotypes was compared. Genotype frequencies of triathletes were compared with the control group as well.Twenty-nine national level Indian Army Triathletes who volunteered for the study were taken as subjects and 101 healthy age matched control group subjects were taken for comparison of genotype frequencies. The following parameters were checked in triathletes: blood pressure, body mass index, VONo difference was observed between fitness parameters of three genotype groups’ triathletes, while the frequency of I allele was found to be very high in triathletes compared with the control group.It can be concluded that there might be a positive association between I allele of ACE gene and endurance. This individual variability in exercise responses has been described as a normal biological phenomenon that may reflect genetic diversity. One of such genetic factors affecting response to training is angiotensin converting enzyme gene polymorphism which has been widely studied , 4. Angiotensin-converting enzyme  is associated with the regulation of blood pressure and maintenance of salt and water homeostasis of the body. Renin which is secreted from juxtaglomerular cells in the kidney acts on angiotensinogen and converts it into angiotensin I. Angiotensin converting enzyme (ACE) converts angiotensin I into angiotensin II, which is the final active material. Such physiological response is predominantly mediated by Angiotensin II specific receptors (AT1 and AT2), located on the cellular surface.It is well known that the effects of exercise training gives inter individual differences. There is a 287 bp insertion (I) / deletion (D) polymorphism in intron 16 of ACE gene that occurs commonly and accounts for a substantial portion of the variance in serum ACE levels. D allele has been associated with a higher ACE activity in the serum and tissue than the I allele.ACE gene (21 Kbp) is located on the chromosome 17 q23 and is composed of 26 exons, 10–12 and D allele with greater strength, 14. Yet, there have been other studies which have found no association between these variables–17.Some studies have associated I allele with greater endurance2max), power, muscular endurance, flexibility and body mass index (BMI).Several studies have been done in various parts of the world but not on Indian population. We have made an attempt to bridge this gap and contribute to the worldwide data being generated on the association between ACE polymorphism and physical fitness phenotypes. The purpose of this study was to find if any association exists between the elite endurance athlete status and ACE polymorphism in Indian population. We also wanted to find whether any difference exists between the three genotype groups of the endurance athletes concerning the physical fitness parameters, such as maximum volume of oxygen uptake  allele and that of a 490 bp fragment indicated the presence of Insertion (I) allele. The individuals homozygous for the insertion allele were designated as II, heterozygous as ID and those homozygous for deletion allele were designated as DD. In order to increase the specificity of DD genotyping, all samples identified as DD after initial amplification were reconfirmed with an insertion-specific primer pair.. The genotypes of all subjects were recorded.The presence of insertion sequence was revealed by the amplification of a 275 bp fragment, while DD homozygotes failed to be amplified due to the lack of annealing site2 max:Measurement of VO VO2 max measurements were taken using Queen's college step test.Procedure followed for step test: A 16.25 inches high stepper was used. Subjects had to step up and down for 3 min at a rate of 24 steps per min. Rate was regulated using a metronome. The subject immediately stopped on completion of the test, and the heart beats were counted for 15 seconds from 5-20 seconds of recovery and recorded. The following formula was used to calculate VO2max:Measurement of muscular endurance: Muscular endurance was assessed using one minute push-up and curl-up test.Procedure for push-up test: Subjects performed this test in the standard ‘on toes’ position. The subject lowered the body until the chin touched the mat (It was instructed that abdomen should not touch the mat). Subject was reminded to keep his back straight at all times and to push up to a straight-arm position. The maximum number of push-ups performed consecutively without rest was counted.Procedure for Curl-ups: The subject assumed a supine position with knees bent so that the heels are positioned approximately 18 inches from the buttocks. The arms were held at the side, with a strip of masking tape placed on the floor at the fingertips. A second strip of tape was placed exactly 12 cm beyond the first strip (towards heels). The subject performed slow, controlled curl-ups, lifting the inferior border of scapula off the mat and returning to the start position after each repetition. To successfully complete a repetition, the subject had to touch the fingertips to the second strip of tape, with the trunk making a 30-degree angle with the mat and the low back flattened before each curl-up. At the ‘go’ signal, the subject performed as many curl-ups as possible in 1 min.Measurement of muscle strength and power: Strength of upper body was assessed using push-ups. Vertical jump was used to assess lower body strength.Harman formula was used to convert vertical jump scores into peak power and average power.Procedure: A measuring tape and chalk for marking the wall was used. The athlete stood with side on to a wall and reached up with the dominant hand closest to the wall. Keeping the feet together and flat on the ground, the point of the fingertips was marked and recorded as the ‘standing reach’. The athlete then performed the vertical jump using the countermovement jump technique described by Adams. Each subject was then given three opportunities to jump. Shuffling of feet or steps was not allowed. The difference between the standing reach and the higher jump to the nearest 1cm was recorded as their vertical jump. The difference in distances between the standing reach height and the jump height was the score. The best of three attempts was recorded. Later jump height was converted into a power score.Vertical Jump Power Calculation: We have used the Harman Formula. Harman et al established equations for both peak and average power through multiple regression procedures. The two equations are listed below:Measurement of flexibility: Flexibility was assessed using modified sit and reach test.This test measures the flexibility of the lower back and hamstring muscles.Procedure: Three trials were given following a warm-up. The test involved sitting on the floor with back and head against a wall, legs fully extended, with the bottoms of the feet (shoes off) against the sit-and-reach box. Hands were placed on top of each other, stretching the arms forward while the head and back kept against the wall. The distance from the fingertips to the box edge was measured with a yardstick. This represented the zero, or starting point. Subject was asked to slowly bend forward and reach as far as possible, sliding the fingers along the yardstick; the final position was held for 2 seconds. Total distance reached to the nearest ¼ inch represented the final score.BMI: BMI was measured using height and weight.Population studied and sample size: The study subjects were volunteers, 29 male national level Indian army Triathletes with their age ranging from 20 to 25 years (after permission of the authorities) and 101 healthy aged matched subjects as a control group (for comparing genotype frequencies). The sample has been selected from Triathalon that is an event which requires very high endurance. Hence, we wish to study the relation between physical fitness and ACE polymorphism. The subjects belonged to heterogenous genotype groups, while being exposed to similar training and food.2max, peak power, average power, number of curl-ups, number of push-ups and score on sit and reach test between the three groups. Consequently, a Post Hoc Tukey's HSD Test was applied. The two groups (triathletes and controls) were compared according to allele and genotype frequencies by using χ2 analysis.SPSS version 17 was used for analysis of the data. Levene's test was applied to check the homogeneity of variance. One- way ANOVA was used to compare all variables including BMI, VOP>0.05)  .2 =5.36, df:1, P=0.02). The same trend was found in the genotypic distributions, as triathletes showed an excess of the II genotype . Distribution of I and D alleles and their genotypes for the control subjects are listed in Distributions of I and D alleles and ACE genotypes are given in Genotype frequencies were in Hardy Weinberg equilibrium in both triathletes and controls, making selection bias less likely. Our results indicate I allele dominance in triathletes compared with the control group. On the other hand, no statistically significant difference was found between the three genotypes  in any of the physical fitness parameters measured.2 max:ACE and VO There was no statistically significant difference between the VO2 max of triathletes of the three genotype groups (2 max of three genotypes triathletes. The sample size of our study is also too small to be able to make such a definite assumption. Various studies have suggested an association between ACE I/D polymorphism and fitness parameters. A significant excess of II and ID ACE genotypes was demonstrated in a study involving 25 elite high-altitude British mountaineers. Moreover, a positive association was found between I allele and endurance performance among British Olympic standard runners . In another study done on postmenopausal women ACE II genotype carriers had a greater VO2max than ACE ID or ACE DD genotype carrier. Montogmery et al. also reported that I allele is associated with improved endurance capability. Hypoxia induced increased ventilation was shown to be significantly greater among type II genotype carriers. Tsianos and colleagues investigated the frequency of the ACE I/D polymorphism among 35 elite endurance swimmers. They found an excess of I allele in swimmers participating in 25-km races requiring more endurance and a significant excess of D allele among subjects competing in 1- to 10-km races, which may require more strength or power. This may suggest a relation between duration of event and I/D polymorphism, with I allele favouring long duration events requiring endurance.e groups  while a e groups . This ma, 17, 26 have failed to show effects of ACE genotype on VO2 max or clustering of ACE genotype II in elite athletes participating in sports with high aerobic fitness demands, 16, 29. Additionally, a recent study by Bouchard et al. found no linkage between chromosome 17genetic markers, location of the ACE gene, and either baseline VO2 max or the VO2 max response to training.Other recent investigations. This may explain different responses shown by different studied population groups. Furthermore, although VO2 max is clearly important for aerobic events performance, it only accounts for 80-90% of the variance in subjects’ running times, while lactate threshold and running economy account for most of the rest. This may explain why no association was found in some studies.The effect of ACE genotype may be eclipsed in young adults by the importance of other biological factors, such as the hormonal environment, 33. Overall, the preponderance of evidence suggests that some sorts of associations exist between ACE genotypes and endurance. Larger intervention based studies on various homogenous population groups are required to evaluate, validate and consolidate such claims.Researches which have linked D allele with endurance are in contrast to our study ACE and Strength or Power: In our study no association was found regarding peak power and average power between three genotypes ( conducted a study on 33 healthy males involving 9 weeks of quadriceps strength training to examine the effect of ACE polymorphism on physical performance and finding greater strength gains in subjects with D- allele. Williams et al. in their study found D allele to be associated with isometric and isokinetic quadriceps muscle strength. Furthermore, D allele has recently been associated with left ventricular hypertrophy in response to physical training. In contrast, Woods et al. in a study on post- menopausal women found that I allele was associated with greater strength gains. In another study ACE ID genotype was found to be associated with the contralateral effects of unilateral resistance training.enotypes . There h that are favoured in power performance. On the contrary our study showed no relation between ACE polymorphism and power. According to the results it can be inferred that either there is no relation or the statistical power of the study is not adequate to detect it. Results may also be confounded by the fact that studied athletes belonged to a high endurance sport.It is interesting to know that angiotensin II may be important in blood flow redirection from type I muscle fibers to type II fibersACE and Muscular endurance: The results obtained in our study showed no significant association between number of performed curl-ups and number of push-ups and any genotype group ( who employed similar tests in their study on US army recruits.pe group . Though ACE and Flexibility: No statistically significant association has been found between genotype groups and flexibility (xibility . This isACE and BMI: We found no statistically significant difference between the genotypes in terms of BMI ( investigated the metabolic effects of the human local renin-angiotensin system within subjects involved in an intensive exercise program and showed that those with II genotype had a greater fat mass anabolic response than did those with either ID or DD genotypes. This obtained data contradicts the findings of Katsuya and colleagues, who found an association between the ACE I allele and an increased body mass index.s of BMI , althougA significant strength of our study was the fact that a healthy age matched control group was taken for comparison of genotype frequencies. Besides, high motivation levels of army triathletes ensured that they performed their best on all the tests.One of the limitations of our study was the fact that a heterogenous population was enrolled. Vast genetic dissimilarity can be considered as a potential confounding factor of the findings. Due to practical limitations field tests, which are not as sensitive as direct lab tests, were employed for measuring various physical fitness parameters. However, as direct lab tests are more sensitive, they should be used in future studies to improve the reliability of obtained results.Furthermore, the very small sample size of triathletes which was available for the present study is not enough to generalise the findings. A larger study done at different centres across the country can take the advantages of a small sample of a homogenous group and yet the combined result would be applicable for a large heterogeneous population. Similar studies on different population groups in terms of age, physical fitness and disease profile can help in better understanding of the role of ACE in physical performance.Since a positive association is found between genotype and athletic ability, we recommend a larger longitudinal study to screen the children with athletic ability and study their response to training and its association with genetic polymorphism.Based on the findings of this study it can be concluded that there is an association between I allele of ACE and endurance. There may be separate and complementary actions of I and D alleles. The finding of allele I dominance in triathletic group, and even a suggestion of II homozygote advantage is consistent with this idea.We also conclude that there is no association between ACE polymorphism and power, muscular endurance, flexibility and BMI in this group of national level Indian army triathletes.


Objective: To correlate the clinico-pathological aspects of Oral Squamous Cell Carcinoma (OSCC) with risk factors to determine the present status and variations in the profile.Methodology: One hundred patients of OSCC and one hundred age and sex matched controls were selected. Detailed demographic data, regarding age, gender, marital status, ethnicity, religion, socio-economic status along with habits, betel quid, tobacco chewing / smoking, alcohol and dietary habits was recorded. Detailed oral examination was carried out for the site of involvement and associated pathology. Histological grade was determined on microscopic examination of Hemotoxylin & Eosin (H&E) stained slides. One hundred age and sex matched controls were also evaluated for this study.Results: Ages of patients ranged from 25 to 80 years with mean age being 47.84 ± 12.18(SD). Maximum cases were detected in the fifth decade. Male: Female ratio was 2.8:1. Age in controls ranged from 22 -73 with male to female ratio being 3.54:1. In patients, most tumors were seen in buccal cavity (54%) followed by tongue (24%). Histologically 60% cases were well differentiated. Strong association with tobacco smoking and chewing, betel quid and its substitutes was detected, with smoking being more prevalent in males and betel quid in females. Significantly less number of controls were observed to be involved in these habits, with almost half having no such addictions.Conclusion: The present clinico-pathological status of oral cancer still emphasizes primary prevention by creating awareness against the devastating effects of tobacco use, betel quid, its substitutes and areca nut, which can go a long way in decreasing the incidence of this disfiguring and lethal condition. It is a cause of great concern all over the world and a major threat to public health in Pakistan, though a wide geographical variation in incidence and mortality is observed. It is the eighth most common malignancy internationally but in Pakistan it is the second commonest as per recent records of an established and well maintained cancer registry of Shaukat Khanum Memorial Hospital.4Most cases have been reported in middle and older age groups but in recent years a number of studies have shown earlier age of incidence. More than 90% of cases of oral cancer are Squamous cell carcinomas (OSCC).5OSCC has a multifactorial etiology with contributions of both genetic and environmental influences, suggesting an overwhelming role of the latter. Majority of the cases are attributable to separate and combined habits of tobacco use (smoked and chewed), alcohol consumption, betel quid, areca nut and betel quid substitutes. A number of recent studies are confirming the putative role of Human Papilloma Virus, especially in the western world, which still needs to be investigated in our population.6,7 Developing Asian countries present a different scenario with greater prevalence of tobacco, betel quid, areca nut and its substitutes as major carcinogenic influences.8Tobacco and alcohol are globally accepted and well documented strongest risk factors, most prevalent in the western countries with a multiplicative synergistic effect that has been shown in a number of studies.9 Cigarettes are the most common of the various forms of smoked tobacco. Smokers have 27 fold more chances of developing OSCC in comparison with non-smokers. Reduction or cessation in smoking leads to a decrease in its incidence and mortality.10Almost 90% of all oral cancers are caused by tobacco.11Nitrosamines are again the most important group of carcinogens in smoked and chewed tobacco. Susceptibility of a person depends not only on dose but also on uptake, metabolism, activation and detoxification of these substances. Chewable tobacco increases the risk of OSCC in proportion to its frequency and duration of use.12 Betel quid (Paan) chewing, on the other hand is a socially and culturally supported habit in Pakistan and rest of Central and South East Asia.13 Areca nut is highly addictive and chronic use leads to Sub mucus fibrosis and OSCC due to its mutagenic and genotoxic effects mediated by areca nut alkaloids and free radicals. Their easy availability, in form of cheap, attractive sachets is responsible for popularizing its use in our country in all age groups, even in primary school children.14In our country alcohol use is abhorred as it is not available in open market due to religious and social influences.15Betel quid substitutes (BQS) like gutka, pan masala, etc which are becoming popular in Indo- Pakistan are linked to a higher genotoxic and carcinogenic potential with major contributions from tobacco nitrosamines and areca nut alkaloids.Frequently accompanying poor oral hygiene with chronic irritative sepsis can also contribute to the carcinogenesis of established risk factors by elaboration of free radicals and cytokines. Nitrosamine formation may be augmented by the bacterial enzymes.16 Therefore vegetables and fruits have been shown to have a beneficial effect, though it may be attributable to vitamins found in abundance in these fruits.7Epidemiological studies have described a role of hot and spicy foods and irritants in oral cancer development. Moreover deficiencies of vitamins A,C and E and micronutrients as selenium, folates are associated with increased risk of malignant transformation.Rationale: Numerous studies have been carried out in Western countries on etiological risk factors and their association to clinico-pathological aspects of Oral Cancer in order to determine the pathways of carcinogenesis. OSCC is much more common in Pakistan with a very high incidence and mortality and established risk factors which need to be re-evaluated due to changing lifestyle and hence risk exposure patterns.This is a cross sectional study. It was carried out on 100 cases of clinically diagnosed and histopathologically proven cases of OSCC. The cases were collected from July 2009 – June 2012 from ENT ward of Civil Hospital, Karachi. Hundred age and sex matched controls were also selected from relatives and attendants accompanying the patients to correlate the etiologic risk factors. After informed consent all the patients were interviewed and examined in detail for local pathology.The information regarding their age, gender, marital status, ethnicity, religion, socio-economic status and habits, especially betel quid, betel nut chewing, betel quid substitutes like gutka and pan masala, tobacco, naswar chewing, smoking and alcohol was recorded on a proforma. Dietary habits were also taken into account. Detailed general and local clinical examination was carried out along with condition of oral hygiene and dental status. The types of smoked and chewed products were noted but information regarding duration and frequency was subject to recall bias and variability over time.The tissues obtained at time of biopsy or surgery were fixed in 10% neutral buffered formalin and sent to DDRRL for routine histopathological diagnosis. The representative sections were subjected to routine embedding, processing and staining with Hemotoxylin & Eosin (H&E).The cases of OSCC were graded into Well Differentiated, Moderately and Poorly Differentiated tumors. SPSS version 16 was used for all statistical calculations.to female ratio being 2.84:1. Among controls 78 were males and 22 were females with ages ranging from 22 -73 years. Cheek  was the most common site involved in OSCC patients in both sexes, followed by tongue and alveolar sulcus , followed by Sindhis (28%), Pushtoons (15%), Punjabis (12%) and Baluchis (8%).Information regarding risk factors revealed tobacco smoking, chewing, betel quid and its substitutes to be extensively associated with oral cancer cases. Minimal number of patients showed no habit association. Only one patient admitted to be habituated for taking alcohol. Tobacco smoking alone or in combination with other habits was dominantly found in males whereas BQ was determined as the principal risk factor in females. Betel quid use was associated with tobacco in almost all cases . Majorit17 In our study the maximum numbers of cases were detected in younger age mostly affecting 41-50 years age group. This contrasts with older studies by Mirza et al18 showing maximum number of cases in slightly advanced age of 51-60 years. Study by Bhurgri et al12 in 2006 shows oral cancer in young adults conforming to the change in pattern observed in this present study.12,18 The reason seems to be attributable to the earlier exposure of our population to risk factors like betel quid, areca nut and its substitutes. These are popular in all age groups in both sexes due to attractive presentation, cheap prices and easy availability supported by lack of knowledge about its devastating effects. This trend for oral cancer development in younger ages is noted in the Western world as well but the cases are mostly attributable to oral HPV infection which is more prevalent there due to promiscuous sexual habits.19 Present study shows male predominance with male: female ratio of 2.82:1. Majority of research all over the world reveals a general male dominance.20OSCC is found to be more common in slightly older males all over the world but newer studies are revealing its development in younger age groups.13 The most common site involved in oral cancer cases of this study was the buccal mucosa (cheek), followed by tongue which are in accordance with the risk factors rampant in our region of the world as supported by Ayaz B.21 The chewable forms of tobacco, betel quid, areca nut and gutka are kept in the buccal pouch for long periods and its juices are sucked in. This leads to greater exposure of the subjected mucosa to carcinogens in these habit forming substances. Western countries show tongue to be the commonly affected site in oral cancers followed by floor of mouth as their tobacco exposure is more related to smoking rather than chewing.22The customary risk factors as tobacco, betel quid and its substitutes show a significant association with OSCC in our subjects, conforming to the previous literature available from Indo-Pakistan Subcontinent . Recent 23, but only one case of habitual alcohol intake was detected in current study. In our country alcohol intake is a taboo and it is possible that its full disclosure may have been subject to this bias and the possibility of hidden exposure in a few cases cannot be excluded.Alcohol is a major risk factor in the Western regions17 The present study also showed a deficient intake of fruits and vegetables in oral cancer patients. Fruits and vegetables may exert their influence indirectly through antioxidant effect of various nutrients as Vitamins A, C and E and micronutrients as iron, folic acid etc., contained therein. Independent assessment of these vitamins and micronutrients was beyond the scope of this study and requires a comprehensive epidemiological survey; however supportive evidence is still generated. Furthermore most patients evaluated belonged to the lower social strata. This could further have had a bearing on low intake of fruits and vegetables in this class which can barely afford two square meals a day for the family.Role of other risk factors like fruits and vegetables has been investigated which shows to have an inverse relationship with oral carcinogenesis.24Effects of poor dentition and oral sepsis have shown equivocal results on comparing cases and controls in the present study. Detailed appraisal with microbiologic cultures for polymicrobial supragingival infections is advocated as chronic inflammation with cytokines could obviously fuel the fire for carcinogenic agents.Outcomes of the present study revealed differences in the risk factor exposure pattern between genders and various age groups with tendency for oral cancer development in younger ages. The aggressive role played by tobacco, betel quid, and areca nut and betel quid substitutes in our population reinforces the commitment for primary prevention and strong implementation of rules to curb these disguised social evils prevalent in our society at large, with immediate effect.There is a dire need for creating a campaign regarding awareness in all age groups, especially children against the use of these addictive carcinogenic substances, in order to bring down the incidence of this aggressive, disfiguring, and rapidly fatal condition.SA conceived, designed, acquired and analyzed data and did writing of manuscript.TM designed and did review, editing and final approval of manuscript.MA did data collection and reviewed and did final approval of manuscript.MQ did review and final approval of manuscript.


Deficits in recalling the past and imagining fictitious and future scenarios have been documented in patients with hippocampal damage and amnesia that was acquired in adulthood. By contrast patients with very early hippocampal damage and developmental amnesia are not impaired relative to control participants when imagining fictitious/future experiences. Recently, however, a patient (HC) with developmental amnesia, resulting from bilateral hippocampal atrophy, was reported to be impaired, thus raising a question about the true nature of event construction in the context of developmental amnesia. Here, we assessed HC on a test of imagination which explored her ability to construct fictitious events or personal plausible future events. Her scenario descriptions were analysed in detail along a range of parameters, using two different scoring methods. HC's performance was comparable to matched control participants on all measures relating to the imagination of fictitious and future scenarios. We then considered why she was reported as impaired in the previous study. We conclude that various features of the previous testing methodology may have contributed to the underestimation of HC's ability in that instance. Patients like HC with developmental amnesia may be successful at future-thinking tasks because their performance is not based on true visualisation or scene construction supported by the hippocampus, but rather on preserved world knowledge and semantic representations. One study by n = 21) who had experienced neonatal hypoxia/ischaemia, and consequent bilateral hippocampal damage of varying extent. They too were able to successfully construct fictitious scenarios, despite being impaired at recalling recent autobiographical events. This was the case even for the patients with the most severe memory impairment and the greatest hippocampal volume loss. These results from the developmental patients are in clear contrast to impaired scene construction in patients with adult-acquired hippocampal damage. It has been suggested that a relatively intact semantic memory store and/or some functionality in residual hippocampal tissue may underpin this intact scene construction ability in DA patients  has been examined. Such patients sustain selective bilateral hippocampal damage as a result of hypoxic/ischaemic episodes that occur perinatally or early in childhood. They typically have intelligence in the normal range, and relatively preserved semantic memory, but are amnesic for recalling autobiographical events and have impaired spatial navigation . Maguirepatients .Findings to date seem to suggest a difference in the effects of hippocampal pathology on future-thinking depending on whether damage occurs in adulthood or much earlier in life. However, another DA patient, HC DA6 in  was receWe did this by testing HC when she visited London. We administered the scene construction test used in our previous studies of adult-acquired hippocampal damage  and DA . On the 22.1HC is right-handed, female, a native English-speaker, and was 22 years old at the time of testing. She was born prematurely at 32 weeks of gestation, and experienced significant respiratory distress requiring intubation and ventilation as a neonate. After discharge from the hospital, HC achieved her developmental milestones normally. Memory difficulties and motor coordination problems were noted around the age of 4 years, before school entry. Throughout her primary and secondary education, HC attended mainstream schools, successfully graduating from high school; she went on to complete a year in technical college, and is currently working towards a culinary degree.n = 12) of comparable age were 48.1% on the left, and 43.4% on the right (n = 65) were 45.3% on the left, and 44.8% on the right . All participants gave informed written consent to participation in the study in accordance with the local research ethics committee.Fourteen healthy females, who were native English-speakers, and matched to HC on age, IQ (Wechsler Test of Adult reading – WTAR) and handedness also participated in the experiment. The mean age of the control group was 21.5 years  and mean estimated full-scale IQ was 108.5 . There was no significant difference between the control participants and HC on either of these background characteristics  and the participant was instructed to vividly imagine the situation suggested by the cue and describe it in as much  detail as possible. Participants were explicitly told not to recount an actual memory or any part of one but rather create something new. A printed text card was placed on the desk in front of the participant containing the cue sentence to act as a reminder if needed. Participants were allowed to continue with their descriptions until they came to a natural end or they felt nothing else could be added. A probing protocol dictated the appropriate use of statements used by the examiner during the session. These mostly took the form of general probes encouraging further description , or asking for further elaboration on a theme introduced by the participant . It was strictly prohibited for the examiner to introduce any concept, idea, detail or entity that had not already previously been mentioned by the participant. After each scenario, participants were asked to rate their constructions on a number of different parameters  and perceived salience/vividness .Spatial Coherence Index: As part of the feedback on each scenario, participants were presented with a set of 12 statements each providing a possible qualitative description of the newly constructed experience. Participants were instructed to indicate the statements they felt accurately described their construction. They were free to identify as many or as few as they thought appropriate. Of the 12 statements, 8 were ‘integrated’ and indicated that aspects of the scene were contiguous  and 4 were ‘fragmented’ and indicated that aspects of the scene were not contiguous . One point was awarded for each integrated statement selected and one point taken away for each fragmented statement. This yielded a score between −4 and +8 that was then normalised around zero to give final Spatial Coherence Index score ranging between −6  and +6 (completely integrated). Any construction with a negative Spatial Coherence Index was considered to be incoherent and fragmented.Quality judgement: The final scoring component was the scorer's assessment of the overall quality of the construction. Scorers were instructed to rate how well they felt the description evoked a detailed ‘picture’ of the experience in their own mind's eye. Quality ratings could range from 0 (indicating the construction was completely devoid of details and with no sense of experiencing) to 10 (indicating an extremely rich and highly evocative construction that appeared to emerge from an extremely vivid imagining).Several other ratings were also taken. After imagining each new experience, participants rated how difficult they found this on a scale from 1 to 5 (1 – very easy … 5 – very difficult). They were also asked to rate on a scale of 1 to 5 its similarity to an actual memory, in whole or in part .α = 0.97).A series of scenarios (15%) were randomly chosen and scored blindly by a second scorer. To measure inter-rater reliability, the Cronbach's Alpha test of internal consistency was performed on the two raters’ scores, which revealed excellent inter-rater reliability over all sub-components, both with and without the quality judgement included .Each scenario also received qualitative ratings from the scorer for episodic richness (0–6), time (0–3), place (0–3), perception (0–3), and emotion/thoughts (0–3)  −0.18, p = 0.86), entities present (t(13) −1.01, p = 0.33), sensory descriptions (t(13) −1.15, p = 0.27) and thoughts/emotions/actions (t(13) −0.63, p = 0.54).Analysis of uncapped measures of content  also showed that HC's scores were unimpaired in all four categories–spatial references (t(13) 0.33, p = 0.75), showing this subjective scorer rating did not influence the Experiential Index significantly.HC's overall quality judgement score (given by the scorers) was not significantly different from the controls see . When thHC's score on the Spatial Coherence Index was also indistinguishable from controls see .t(13) 0.30, p = 0.77). This was also the case for the Spatial Coherence Index  There were two types of scenario, fictitious and personal future. Mean Experiential Index scores for the fictitious experiences  were not significantly different (t(13) −1.43, p = 0.18). This was also the case for the Spatial Coherence Index . Mean Experiential Index scores for the personal future experiences  were also not significantly different . In the case of the future scenarios, the cues were non-specific . There was no time limit, although typically no trial lasted beyond 5 min. Scoring was as described in 4.2Perhaps the difference in trial number is the reason for the discrepancy between our findings and those of The next obvious difference between the two tasks is how they were scored. Another difference between the two tasks that may be particularly critical is the nature of the cues. While the cues for our general fictitious scenarios typically contained a specific spatial context and primary content cues, this was clearly not the case for our future-oriented scenarios. In those trials the cues did not contain any spatial context or any key content cues and were clearly non-specific . Not only does the participant have to generate a specific scenario idea, but she then has to construct it, and subsequently elaborate upon it. Nevertheless HC was still able to perform comparably to the control participants on these non-specific cue trials. Incidentally, if the argument is that provision of specific cues somehow helps to circumvent the construction problem in hippocampal amnesia, then why are patients with bilateral hippocampal damage and amnesia acquired in adulthood so impaired on our task ? The speInstead, we suggest that the key issue may be with 4.3HC was unimpaired at the construction of fictitious and future experiences using our paradigm. This finding is consistent with other reports in the literature of patients with early hippocampal damage . It is pTo conclude, in this study we attempted to understand why patient HC, with developmental amnesia, was reported to be impaired at imagining the future , contrar