Insertion of an SVA element in MSH2 as a novel cause of Lynch syndrome.

Germline mutations in the DNA mismatch repair (MMR) genes cause Lynch syndrome (LS). In this study, we identified and characterized a novel SINE-VNTR-Alu (SVA) insertion in exon 12 of MSH2 in an individual with early-onset colorectal cancer and a very strong LS family history. RT-PCR analysis indicated a larger aberrant MSH2 transcript in one of the family members. MSK-IMPACT next-generation sequencing and long-range PCR analyses revealed an insertion in MSH2 exon 12 at the c.1972 position in an antisense orientation. The insertion was further characterized as an SVA element approximately 3 kb in length, belonging to the SVA_F1 family of retrotransposons. This variant also segregated with LS related cancers in four affected family members in this family. Based on this evidence, this MSH2 SVA insertion is considered pathogenic.

An alternative approach to establishing unbiased colorectal cancer risk estimation in Lynch syndrome.

PURPOSE: Biallelic pathogenic variants in the mismatch repair (MMR) genes cause a recessive childhood cancer predisposition syndrome known as constitutional mismatch repair deficiency (CMMRD). Family members with a heterozygous MMR variant have Lynch syndrome. We aimed at estimating cancer risk in these heterozygous carriers as a novel approach to avoid complicated statistical methods to correct for ascertainment bias. METHODS: Cumulative colorectal cancer incidence was estimated in a cohort of PMS2- and MSH6-associated families, ascertained by the CMMRD phenotype of the index, by using mutation probabilities based on kinship coefficients as analytical weights in a proportional hazard regression on the cause-specific hazards. Confidence intervals (CIs) were obtained by bootstrapping at the family level. RESULTS: The estimated cumulative colorectal cancer risk at age 70 years for heterozygous PMS2 variant carriers was 8.7% (95% CI 4.3-12.7%) for both sexes combined, and 9.9% (95% CI 4.9-15.3%) for men and 5.9% (95% CI 1.6-11.1%) for women separately. For heterozygous MSH6 variant carriers these estimates are 11.8% (95% CI 4.5-22.7%) for both sexes combined, 10.0% (95% CI 1.83-24.5%) for men and 11.7% (95% CI 2.10-26.5%) for women. CONCLUSION: Our findings are consistent with previous reports that used more complex statistical methods to correct for ascertainment bias. These results underline the need for MMR gene-specific surveillance protocols for Lynch syndrome.

High-risk family colorectal cancer screening service in Ireland: Critical review of clinical outcomes.

BACKGROUND: We present the 15-year experience of a family colorectal cancer screening service in Ireland with emphasis on real life experience and outcomes. METHODS: Questionnaires were used to assess family cancer history and assign patients to risk categories; 'Moderate Risk', HNPCC, (suspected) genetic syndrome (non-HNPCC), 'Low Risk'. Screening was by full colonoscopy. We report neoplastic yield, examining effect of risk category, age, gender, and index colonoscopy findings. RESULTS: Between 1998 and 2013, 2242 individuals were referred; 57.3% female, 42.7% male, median age 46 years (range9-85yrs). Median follow up time was 7.9yrs (range 0.5-15.3yrs). Follow up data after exclusion (non-compliance, known CRC) was available in 1496 (66.7%): 'Moderate risk' 785 (52.5%), HNPCC 256 (17.1%), (suspected) genetic syndrome (non-HNPCC) 85 (5.7%), 'Low Risk' 370 (24.7%). Screening was performed in 1025(68.5%) patients; colonoscopy data available for 993 (96.9%); total 1914 colonoscopies. At index colonoscopy, 178 (18.0%) patients had adenomas; 56 (5.5%) advanced adenoma. During the entire study period, 240 (24.2%) had an adenoma; 69 (7.0%) advanced adenoma. Cancers were diagnosed on screening in 2 patients. Older age and male gender were associated with higher adenoma detection rate; p<0.001, p=0.01, respectively. Risk category did not affect adenoma yield. Adenoma and advanced adenoma detection at index colonoscopy were associated with detection of same at follow up screening; p<0.001. CONCLUSION: Male gender and age (>50) were the core identifiable risk factors for neoplasia at screening colonoscopy in this family screening setting. Our results would support less intensive surveillance in younger patients (<50), particularly where index colonoscopy is normal.

Application of a 5-tiered scheme for standardized classification of 2,360 unique mismatch repair gene variants in the InSiGHT locus-specific database.

The clinical classification of hereditary sequence variants identified in disease-related genes directly affects clinical management of patients and their relatives. The International Society for Gastrointestinal Hereditary Tumours (InSiGHT) undertook a collaborative effort to develop, test and apply a standardized classification scheme to constitutional variants in the Lynch syndrome-associated genes MLH1, MSH2, MSH6 and PMS2. Unpublished data submission was encouraged to assist in variant classification and was recognized through microattribution. The scheme was refined by multidisciplinary expert committee review of the clinical and functional data available for variants, applied to 2,360 sequence alterations, and disseminated online. Assessment using validated criteria altered classifications for 66% of 12,006 database entries. Clinical recommendations based on transparent evaluation are now possible for 1,370 variants that were not obviously protein truncating from nomenclature. This large-scale endeavor will facilitate the consistent management of families suspected to have Lynch syndrome and demonstrates the value of multidisciplinary collaboration in the curation and classification of variants in public locus-specific databases.

Clinical correlation and molecular evaluation confirm that the MLH1 p.Arg182Gly (c.544A>G) mutation is pathogenic and causes Lynch syndrome.

Approximately 25 % of mismatch repair (MMR) variants are exonic nucleotide substitutions. Some result in the substitution of one amino acid for another in the protein sequence, so-called missense variants, while others are silent. The interpretation of the effect of missense and silent variants as deleterious or neutral is challenging. Pre-symptomatic testing for clinical use is not recommended for relatives of individuals with variants classified as 'of uncertain significance'. These relatives, including non-carriers, are considered at high-risk as long as the contribution of the variant to disease causation cannot be determined. This results in continuing anxiety, and the application of potentially unnecessary screening and prophylactic interventions. We encountered a large Irish Lynch syndrome kindred that carries the c.544A>G (p.Arg182Gly) alteration in the MLH1 gene and we undertook to study the variant. The clinical significance of the variant remains unresolved in the literature. Data are presented on cancer incidence within five kindreds with the same germline missense variant in the MLH1 MMR gene. Extensive testing of relevant family members in one kindred, a review of the literature, review of online MMR mutation databases and use of in silico phenotype prediction tools were undertaken to study the significance of this variant. Clinical, histological, immunohistochemical and molecular evidence from these families and other independent clinical and scientific evidence indicates that the MLH1 p.Arg182Gly (c.544A>G) change causes Lynch syndrome and supports reclassification of the variant as pathogenic.

Silver-enhanced in situ hybridization for detection of polyomavirus DNA in patients with BK virus nephropathy.

BK virus nephropathy is not an infrequent complication of renal transplantation associated with high rates of graft loss. Although antibodies against SV40 antigen detect different viruses of the polyomavirus family, immunohistochemistry is widely used to confirm the diagnosis of BK virus nephropathy in renal biopsies. Here we aimed to validate the novel silver-enhanced in situ hybridization (SISH) technique for the automated detection of BK virus in renal transplant biopsies. Two different patient cohorts were included. Twenty-nine consecutive patients suspicious for BK virus infection were investigated by SISH and chromogenic in situ hybridization. An additional 26 renal biopsies positive by SV40 immunohistochemistry from 19 patients were analyzed by SISH. Polyomavirus DNA serum levels, as determined by nested PCR analysis, were available for all of these patients. The presence of BK virus DNA in renal tubular cells was identified in 5 of the suspicious cases by both, SISH and chromogenic in situ hybridization . One additional patient was only positive in the SISH. In the second cohort, SISH was positive in all SV40 positive biopsies, but SISH signals were less extensive than SV40 immunohistochemistry. Our results show that the BK virus SISH is an ancillary tool for the detection of polyomavirus DNA in renal biopsies using bright-field microscopy. However, its diagnostic value in comparison with standard immunohistochemistry seems to be limited.

Athletic participation and seatbelt omission among u.s. High school students.

Although seatbelts save lives, adolescents may be disproportionately likely to omit their use. Using data from the 1997 Youth Risk Behavior Survey, a national survey of more than 16,000 U.S. public and private high school students, the authors employed a series of logistic regression analyses to examine cross-sectional associations between past year athletic participation and regular seatbelt omission. Controlling for the effects of gender, age, race, parental education, and school urbanicity, student athletes were significantly less likely than nonathletes to report seatbelt omission. Separate gender-specific analyses showed that this effect was significant for girls but only marginally significant for boys; in addition, the effect was strongest for adolescents who participated on three or more school or community sports teams. Possible explanations for the relationship between athletic participation and seatbelt omission, including Jessor's problem behavior syndrome, prosocial sport subcultures, and sensation seeking, are considered.

Athletic Involvement and Adolescent Delinquency.

Athough conventional wisdom suggests that organized sport deters delinquency by building character, structuring adolescents' time, and providing incentives for socially approved behavior, the empirical evidence to date has been mixed. Based on a sample of approximately 600 Western New York adolescents, the present study examined how self-reported jock identity, school athlete status, and frequency of athletic activity differentially influenced a range of delinquent behaviors. Neither athlete status nor frequency of athletic activity predicted these behaviors; however, jock identity was associated with significantly more incidents of delinquency. This finding was robust across both gender and race. Follow-up analyses indicated that jock identity facilitated both minor and major delinquency, with major delinquency effects for white but not black adolescents.

Jocks, gender, binge drinking, and adolescent violence.

Previous research has suggested a link between athletic involvement and elevated levels of adolescent violence outside the sport context. The present study expanded on this literature by positing differences in the sport-violence relationship across dimensions of athletic involvement (athletic participation vs. jock identity), type of violence (family vs. nonfamily), and gender as well as by examining the impact of binge drinking on the sport-violence relationship. Regression analyses using a sample of 608 Western New York adolescents indicated that (a) jock identity (but not athletic participation) was associated with more frequent violence, (b) jock identity predicted nonfamily violence (but not family violence), and (c) the link between jock identity and nonfamily violence was stronger for boys than for girls. Binge drinking predicted family violence among nonjocks only.

Adolescent anabolic steroid use, gender, physical activity, and other problem behaviors*.

To test the comparative value of strain theory and problem behavior theory as explanations of adolescent anabolic steroid use, this study examined gender-specific relationships among steroid use, physical activity, and other problem behaviors. Based on the United States Centers for Disease Control and Prevention's 1997 Youth Risk Behavior Survey, a nationally representative sample of over 16,000 U.S. public and private high school students, binge drinking, cocaine use, fighting, and sexual risk-taking were associated with higher odds of lifetime steroid use. In gender-specific analyses, steroid use was strongly associated with female fighting and smokeless tobacco use as well as male sexual risk. Neither athletic participation nor strength conditioning predicted odds of steroid use after controlling for problem behaviors, nor did steroid-using athletes report more frequent use than steroid-using nonathletes. The study's limitations and policy implications were noted. These data suggest that other problem behaviors such as substance use, fighting, and sexual risk are better predictors of adolescent steroid use than physical activity. Interventions to prevent steroid use should not be limited to male participants in organized sports programs, but should also target adolescents identified as at risk for other problem behaviors.

HIGH SCHOOL ATHLETIC PARTICIPATION AND ADOLESCENT SUICIDE: A Nationwide US Study.

Suicide is the third leading cause of death among US adolescents aged 15-24, with males incurring higher rates of completion than females. This study used hierarchical logistic regression analysis to test whether athletic participation was associated with lower rates of suicidal ideation and behavior among a nationally representative sample of over 16,000 US public and private high school students. Net of the effects of age, race/ethnicity, parental educational attainment, and urbanicity, high school athletic participation was significantly associated with reduced odds of considering suicide among both females and males, and reduced odds of planning a suicide attempt among females only. Though the results point to favorable health outcomes for athletes, athletic participation was also associated with higher rates of injury to male athletes who actually attempted suicide.

Untangling the Links among Athletic Involvement, Gender, Race, and Adolescent Academic Outcomes.

Although previous research has established that high school sports participation may be associated with positive academic outcomes, the parameters of the relationship remain unclear. Using a longitudinal sample of nearly 600 Western New York adolescents, this study examined gender- and race-specific differences in the impact of two dimensions of adolescent athletic involvement ("jock" identity and athlete status) on changes in school grades and school misconduct over a two-year interval. Female and black adolescents who identified themselves as "jocks" reported lower grades than those who did not, whereas female athletes reported higher grades than female nonathletes. Jocks also reported significantly more misconduct (including skipping school, cutting classes, having someone from home called to the school for disciplinary purposes, and being sent to the principal's office) than nonjocks. Gender moderated the relationship between athlete status and school misconduct; athletic participation had a less salutary effect on misconduct for girls than for boys.

Gender/Racial Differences in Jock Identity, Dating, and Adolescent Sexual Risk.

Despite recent declines in overall sexual activity, sexual risk-taking remains a substantial danger to US youth. Existing research points to athletic participation as a promising venue for reducing these risks. Linear regressions and multiple analyses of covariance were performed on a longitudinal sample of nearly 600 Western New York adolescents in order to examine gender- and race-specific relationships between "jock" identity and adolescent sexual risk-taking, including age of sexual onset, past-year and lifetime frequency of sexual intercourse, and number of sexual partners. After controlling for age, race, socioeconomic status, and family cohesion, male jocks reported more frequent dating than nonjocks but female jocks did not. For both genders, athletic activity was associated with lower levels of sexual risk-taking; however, jock identity was associated with higher levels of sexual risk-taking, particularly among African American adolescents. Future research should distinguish between subjective and objective dimensions of athletic involvement as factors in adolescent sexual risk.

Thymidylate synthase inhibitors as anticancer agents: from bench to bedside.

Thymidylate synthase (TS) is a folate-dependent enzyme that catalyzes the reductive methylation of 2'-deoxyuridine-5'-monophosphate to 2'-deoxythymidine-5'-monophosphate. This pathway provides the sole intracellular de novo source of 2'-deoxythymidine-5'-triphosphate; therefore, TS represents a critical target in cancer chemotherapy. 5-Fluorouracil (5-FU) was synthesized in 1957 and represents the first class of antineoplastic agents to be developed as inhibitors of TS. While 5-FU has been widely used to treat various human malignancies, its overall clinical efficacy is limited. Therefore, significant efforts have focused on the design of novel, more potent inhibitor compounds of TS. These agents fall into two main categories: folate analogs and nucleotide analogs. Five antifolate analogs are currently being evaluated in the clinic: raltitrexed, pemetrexed, nolatrexed, ZD9331, and GS7904L. Our laboratory has identified a novel mechanism of resistance that develops to TS inhibitor compounds, namely drug-mediated acute induction of new TS synthesis; this mechanism is directly controlled at the translational level. The ability of cancer cells to acutely induce the expression of TS may represent a novel mechanism for the development of cellular drug resistance. The future success of TS inhibitor compounds in the clinic may depend on novel strategies to selectively inhibit TS and on novel combination therapies to overcome cellular drug resistance.

Jocks, gender, race, and adolescent problem drinking.

Alcohol remains the drug of choice for many adolescents; however, the nature of the relationship between athletic involvement and alcohol misuse remains ambiguous. In this article, we used a longitudinal sample of over 600 Western New York adolescents and their families to explore the gender-specific and race-specific relationships between identification with the "jock" label and adolescent alcohol consumption, specifically problem drinking. Operationalization of problem drinking included frequency measures of heavy drinking, binge drinking, and social problems related to alcohol (e.g., trouble with family, friends, school officials over drinking). Self-identified adolescent "jocks" were more likely to engage in problem drinking than their non-jock counterparts, even after controlling for gender, age, race, socioeconomic status, physical maturity, social maturity, and frequency of athletic activity. Jock identity was strongly associated with higher binge drinking frequency in Black adolescent girls. This study underscores the need to distinguish between objective and subjective meanings of athletic involvement when assessing the relationship between sport and adolescent health-risk behavior.

Thymidylate synthase: a critical target for cancer chemotherapy.

Thymidylate synthase (TS) is a key enzyme in the synthesis of 2'-deoxythymidine-5'-monophosphate, an essential precursor for DNA biosynthesis. For this reason, this enzyme is a critical target in cancer chemotherapy. As the first TS inhibitor in clinical use, 5-fluorouracil (5-FU) remains widely used for the treatment of colorectal, pancreatic, breast, head and neck, gastric, and ovarian cancers. The reduced folate, leucovorin, has been shown to enhance the activity of 5-FU in colorectal cancer. However, response rates of the combination remain in the 25%-30% range, and much effort has been focused on designing new, more potent TS inhibitors. Raltitrexed is a folate analogue that is approved as first-line therapy for advanced colorectal cancer in Europe, Australia, Canada, and Japan, although it remains an investigational agent in the United States. Pemetrexed is an antifolate analogue that has shown promising activity in several solid tumor types, including mesothelioma. ZD9331, a highly specific TS inhibitor that dose not require polyglutamation for its activation, has shown activity in patients with refractory ovarian and colorectal cancer. Capecitabine is an oral fluoropyrimidine carbamate that was designed to generate 5-FU preferentially in tumor cells; this agent was recently approved by the US Food and Drug Administration as first-line therapy for patients with advanced colorectal cancer. As the number of TS inhibitors available for general clinical use increases, further research is needed to elucidate the critical molecular and biochemical elements that determine the efficacy and tumor specificity of each compound.

AKT2 is frequently upregulated in HER-2/neu-positive breast cancers and may contribute to tumor aggressiveness by enhancing cell survival.

Amplification or overexpression of the HER-2/neu gene in breast cancers is associated with aggressive behavior and resistance to therapeutic regimens. The molecular mechanisms that contribute to therapeutic resistance/survival of HER-2/neu-overexpressing tumor cells have not been well defined. To determine if phosphatidylinositol 3-kinase/AKT signaling contributes to cell survival in HER-2/neu-positive breast cancers, we performed immunohistochemical analyses to evaluate expression of HER-2/neu and AKT in a series of 52 breast carcinomas. Elevated expression of HER-2/neu was found to correlate with overexpression of AKT2 protein and activation of AKT kinase. HER-2/neu-overexpressing breast cancer cell lines were resistant to apoptosis induced by UV treatment and hypoxia, which was suppressed in the presence of the phosphatidylinositol 3-kinase inhibitors LY294002 and wortmannin, indicating a link between AKT activation and stress resistance in HER-2/neu-overexpressing cells. These observations suggest that AKT signaling augments resistance to stress-induced apoptosis in breast cancer cells overexpressing HER-2/neu.

Gender and racial/ethnic differences in predicting adolescent sexual risk: athletic participation versus exercise.

Though often conflated, informal physical exercise and organized athletic participation have very different implications for adolescent sexual risk outcomes. The purpose of this research is to disaggregate strenuous exercise from sports, examine how each is associated with sexual risk, and explain the observed differences using the conceptual lens of cultural resource theory. Using a nationally representative sample of over 16,000 public and private high school students, we employ logistic regression to test hypotheses about the gender-specific and race-specific effects of strenuous exercise and athletic participation on adolescent sexual risk behavior. The results show that both forms of physical activity buffer sexual risk for girls. Strenuous exercise is associated with increased odds of sexual risk for boys. Sports and race interact to influence boys' sexual risk outcomes: Athletic participation is associated with lowered odds of sexual risk for white male adolescents, but heightened odds of sexual risk for black male adolescents.