Chromogenic detection of telomere lengths in situ aids the identification of precancerous lesions in the prostate.

BACKGROUND: Telomeres are terminal chromosomal elements that are essential for the maintenance of genomic integrity. The measurement of telomere content provides useful diagnostic and prognostic information, and fluorescent methods have been developed for this purpose. However, fluorescent-based tissue assays are cumbersome for investigators to undertake, both in research and clinical settings. METHODS: A robust chromogenic in situ hybridization (CISH) approach was developed to visualize and quantify telomere content at single cell resolution in human prostate tissues, both frozen and formalin-fixed, paraffin-embedded (FFPE). RESULTS: This new assay (telomere chromogenic in situ hybridization ["Telo-CISH"]) produces permanently stained slides that are viewable with a standard light microscope, thus avoiding the need for specialized equipment and storage. The assay is compatible with standard immunohistochemistry, thereby allowing simultaneous assessment of histomorphology, identification of specific cell types, and assessment of telomere status. In addition, Telo-CISH eliminates the problem of autofluorescent interference that frequently occurs with fluorescent-based methods. Using this new assay, we demonstrate successful application of Telo-CISH to help identify precancerous lesions in the prostate by the presence of markedly short telomeres specifically in the luminal epithelial cells. CONCLUSIONS: In summary, with fewer restrictions on the types of tissues that can be tested, and increased histologic information provided, the advantages presented by this novel chromogenic assay should extend the applicability of tissue-based telomere length assessment in research and clinical settings.

Chromogenic detection of telomere lengths in situ aids the identification of precancerous lesions in the prostate.

Telomeres are terminal chromosomal elements that are essential for the maintenance of genomic integrity. The measurement of telomere content provides useful diagnostic and prognostic information, and fluorescent methods have been developed for this purpose. However, fluorescent-based tissue assays are cumbersome for investigators to undertake, both in research and clinical settings. Here, a robust chromogenic in situ hybridization (CISH) approach was developed to visualize and quantify telomere content at single cell resolution in human prostate tissues, both frozen and formalin-fixed, paraffin-embedded (FFPE). This new assay ("Telo-CISH") produces permanently stained slides that are viewable with a standard light microscope, thus avoiding the need for specialized equipment and storage. The assay is compatible with standard immunohistochemistry, thereby allowing simultaneous assessment of histomorphology, identification of specific cell types, and assessment of telomere status. In addition, Telo-CISH eliminates the problem of autofluorescent interference that frequently occurs with fluorescent-based methods. Using this new assay, we demonstrate successful application of Telo-CISH to help identify precancerous lesions in the prostate by the presence of markedly short telomeres specifically in the luminal epithelial cells. In summary, with fewer restrictions on the types of tissues that can be tested, and increased histologic information provided, the advantages presented by this novel chromogenic assay should extend the applicability of tissue-based telomere length assessment in research and clinical settings.

The Impact Oncoplastic Reduction Has on Long-Term Recurrence in Breast Conservation Therapy.

BACKGROUND: The use of oncoplastic reduction techniques have many proven benefits over lumpectomy alone in the management of women with breast cancer. The impact it has on tumor recurrence is unclear. The purpose of this review was to evaluate the incidence of recurrence in patients who underwent oncoplastic reduction techniques compared to lumpectomy alone. METHODS: A prospectively maintained database of patients at Emory Hospital who underwent oncoplastic reduction techniques at the time of tumor resection was queried. These patients were compared to a series of patients who had lumpectomy alone over a similar period. For inclusion in the study, patients were at least 10 years since the time of the tumor resection. The main outcome of interest was tumor recurrence. RESULTS: There were 97 patients in the lumpectomy-only group and 95 patients in the oncoplastic reduction group, with an average follow-up of 7.8 years and 8.5 years, respectively. Patients in the oncoplastic group were younger (lumpectomy only, 61.4 years; oncoplastic reduction, 51.6 years; p < 0.001) and had larger tumors (lumpectomy only, 1.1 cm; oncoplastic reduction, 1.6 cm; p < 0.001). Local recurrence was 13 percent in the lumpectomy-only group and 9 percent in the oncoplastic reduction group (p = 0.34), and overall recurrence rates were similar (lumpectomy only, 15 percent; oncoplastic reduction, 24 percent; p = 0.13). Overall, surgical intervention (lumpectomy alone versus oncoplastic reduction) was not associated with local recurrence or any recurrence on univariate and multivariate analyses. CONCLUSION: Despite the oncoplastic reduction patients having a higher risk of recurrence and a more generous tumor resection, the long-term recurrence rates were equivalent when compared to breast-conserving therapy alone. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, III.

Pulp De-epithelialization for Venous Outflow Augmentation after Digital Replantation.

Traumatic amputation injuries account for a substantial portion of emergency department visits. This includes digital amputations that may be considered for replantation. Following surgery, venous congestion is the most common cause of replant failure. To address this, several methods have been proposed to augment venous outflow. In this article, a simple and straightforward method that can be utilized to establish or augment venous outflow in cases of venous insufficiency is described. This method entails de-epithelization of the replanted digit pulp skin with use of postoperative anticoagulation. The area can be further expanded or stimulated to increase bleeding as needed and is allowed to heal by secondary intention. This method allows for reliable venous outflow with relative ease of implementation.

School-age social behavior and pragmatic language ability in children with prenatal serotonin reuptake inhibitor exposure.

Studies examining associations between fetal serotonin reuptake inhibitor (SRI) exposure and child autism spectrum disorder (ASD) diagnoses or delayed language remain mixed and rarely prospectively follow children or employ gold-standard assessments. We prospectively followed a cohort of mother-child dyads from pregnancy through early elementary school (N = 178), and obtained maternal and alternate-caregiver ratings of behaviors related to ASD (N = 137), as well as direct, gold-standard assessments of child ASD symptoms and pragmatic language among dyads who experienced prenatal depression and either took SRIs or were medication free during pregnancy (N = 44). Prenatal SRI exposure was related to maternal ratings of ASD-related behaviors (ß = 0.24 95% confidence interval; CI [0.07, 0.48]), and, among boys, alternative caregiver ratings (males-only ß = 0.28 95% CI [0.02, 0.55], females-only ß = -0.21 95% CI [-0.63, 0.08]). However, results of our direct assessments suggest an association between SRI exposure and reduced pragmatic language scores (ß = -0.27, 95% CI [-0.53, -0.01], but not ASD (Autism Diagnostic Observation Schedule ß = 0.14 95% CI [-0.15, 0.41]; Social Responsiveness Scale ß = 0.08 95% CI [-0.25, 0.40]). These discrepancies point to issues regarding how ASD is assessed, and the possibility that SRIs may be more strongly associated with language or other broader behaviors that coincide with ASD. Larger prospective studies that incorporate thorough, gold-standard assessments of ASD, language, and other ASD-related behaviors are needed.

Examining the Overlap between Autism Spectrum Disorder and 22q11.2 Deletion Syndrome.

22q11.2 deletion syndrome (22q11.2DS) is a genomic disorder reported to associate with autism spectrum disorders (ASDs) in 15-50% of cases; however, others suggest that individuals with 22q11.2DS present psychiatric or behavioral features associated with ASDs, but do not meet full criteria for ASD diagnoses. Such wide variability in findings may arise in part due to methodological differences across studies. Our study sought to determine whether individuals with 22q11.2DS meet strict ASD diagnostic criteria using research-based guidelines from the Collaborative Programs of Excellence in Autism (CPEA), which required a gathering of information from three sources: the Autism Diagnostic Interview-Revised (ADI-R), the Autism Diagnostic Observational Schedule (ADOS), and a clinician's best-estimate diagnosis. Our study examined a cohort of children, adolescents, and young adults (n = 56) with 22q11.2DS, who were ascertained irrespective of parents' behavioral or developmental concerns, and found that 17.9% (n = 10) of the participants met CPEA criteria for an ASD diagnosis, and that a majority showed some level of social-communication impairment or the presence of repetitive behaviors. We conclude that strictly defined ASDs occur in a substantial proportion of individuals with 22q11.2DS, and recommend that all individuals with 22q11.2DS be screened for ASDs during early childhood.

Variation in the oxytocin receptor gene moderates the protective effects of a family-based prevention program on telomere length.

INTRODUCTION: Parent-child relationships with high conflict and low warmth and support are associated with later adverse behavioral and physiological child outcomes. These outcomes include shorter telomere lengths, the repetitive sequences at the ends of chromosomes that have been utilized as a biomarker for chronic stress. Our research group furthered this by exploring telomere length outcomes following a family-based prevention program and identified reduced telomere shortening 5 years post intervention among those originally exposed to nonsupportive parenting and randomized to the intervention condition. However, not all individuals respond equally, and a growing literature suggests genetic sensitivity to one's environment, with variations in the oxytocin receptor gene (OXTR) potentially influencing this sensitivity. METHODS: We utilized data from African American youths (mean age 17) randomized to intervention (n = 100) or control condition (n = 91) with baseline assessments of genetic status and nonsupportive parenting, and 5-year follow-up assessments of telomere length. RESULTS: We found a significant three-way interaction between nonsupportive parenting, intervention condition, and OXTR rs53576 genotype. OXTR GG individuals, who are suggested to be more sensitive to their social environment, exhibited significantly more variability, evidencing the shortest telomeres when exposed to nonsupportive parenting and randomized to the control condition, and similar telomere lengths to non at-risk groups when randomized to the intervention. In contrast, those with the A allele showed no statistical difference in telomere lengths across parental and intervention conditions. Subsequent analyses suggest that these findings may be mediated through chronic anger, whereby GG individuals exposed to nonsupportive parenting and randomized to the control condition had a greater increase in chronic anger by study follow-up, compared to those in the intervention, and this change associated with greater telomere shortening. CONCLUSIONS: These findings highlight the importance of individual differences and potential role of genetic status in moderating the relationship between environmental contexts and biological outcomes.

Oxytocin Receptor Genetic and Epigenetic Variations: Association With Child Abuse and Adult Psychiatric Symptoms.

Childhood abuse can alter biological systems and increase risk for adult psychopathology. Epigenetic mechanisms, alterations in DNA structure that regulate the gene expression, are a potential mechanism underlying this risk. While abuse associates with methylation of certain genes, particularly those in the stress response system, no study to date has evaluated abuse and methylation of the oxytocin receptor (OXTR). However, studies support a role for OXTR in the link between abuse and adverse adult outcomes, showing that abuse can confer greater risk for psychiatric symptoms in those with specific OXTR genotypes. This study therefore sought to (a) assess the role of epigenetics in the link between abuse and psychopathology and (b) begin to integrate the genetic and epigenetic literature by exploring associations between OXTR genotypes and DNA CpG methylation. Data on 18 OXTR CpG sites, 44 single nucleotide polymorphisms, childhood abuse, and adult depression and anxiety symptoms were assessed in 393 African American adults (age = 41 ± 12.8 years). Overall, 68% of genotypes were associated with methylation of nearby CpG sites, with a subset surviving multiple test correction. Child abuse associated with higher methylation of two CpG sites yet did not survive correction or serve as a mediator of psychopathology. However, abuse interacted with CpG methylation to predict psychopathology. These findings suggest a role for OXTR in understanding the influence of early environments on adult psychiatric symptoms.

Genetic sensitivity to emotional cues, racial discrimination and depressive symptoms among African-American adolescent females.

Psychosocial stress, including stress resulting from racial discrimination (RD), has been associated with elevated depressive symptoms. However, individuals vary in their reactivity to stress, with some variability resulting from genetic differences. Specifically, genetic variation within the linked promoter region of the serotonin transporter gene (5-HTTLPR) is related to heightened reactivity to emotional environmental cues. Likewise, variations within this region may interact with stressful life events (e.g., discrimination) to influence depressive symptoms, but this has not been empirically examined in prior studies. The objective of this study was to examine whether variation in the 5-HTTLPR gene interacts with RD to predict depressive symptoms among a sample of African-American adolescent females. Participants were 304 African-American adolescent females enrolled in a sexually transmitted disease prevention trial. Participants completed a baseline survey assessing psychosocial factors including RD (low vs. high) and depressive symptomatology (low vs. high) and provided a saliva sample for genotyping the risk polymorphism 5-HTTLPR (s allele present vs. not present). In a logistic regression model adjusting for psychosocial correlates of depressive symptoms, an interaction between RD and 5-HTTLPR group was significantly associated with depressive symptomatology (AOR = 3.79, 95% CI: 1.20-11.98, p = 0.02). Follow-up tests found that high RD was significantly associated with greater odds of high depressive symptoms only for participants with the s allele. RD and 5-HTTLPR status interact to differentially impact depressive symptoms among African-American adolescent females. Efforts to decrease depression among minority youth should include interventions which address RD and strengthen factors (e.g., coping, emotion regulation, building support systems) which protect youth from the psychological costs of discrimination.

Social stress and the oxytocin receptor gene interact to predict antisocial behavior in an at-risk cohort.

Polymorphisms in the oxytocin receptor gene are commonly associated with prosocial behaviors in the extant literature, yet their role in antisocial behaviors has rarely been explored, particularly during the transition from adolescence to early adulthood. We examined a prospective cohort (N = 404), collecting youth, mother, and clinician reports of conduct-disordered and antisocial behavior at ages 15 and 20. The oxytocin receptor gene rs53576 polymorphism was hypothesized to interact with social stress to predict antisocial outcomes. Structural equation modeling results revealed a significant main effect at age 15 (p = .025); those with the G allele exhibited higher levels of conduct problems. Structural equation modeling revealed a significant Gene × Environment interaction at age 20 (p = .029); those with the G allele who experienced high social stress exhibited higher levels of antisocial behavior. Heterozygous (AG) grouping models were compared, and parameter estimations supported G dominant groupings. These novel findings suggest that rs53576 polymorphisms may influence social salience and contribute to risk for antisocial outcomes, particularly under conditions of high social stress.

Associations Between a Dopamine D4 Receptor Gene, Alcohol Use, and Sexual Behaviors among Female Adolescent African Americans.

Adolescent African-American females are disproportionately impacted by HIV, thus there is a clear need to understand factors associated with increased HIV-risk behaviors among this vulnerable population. We sought to explore the association between a dopamine D4 receptor gene (DRD4), a genetic marker associated with natural variations in rewarding behaviors, and self-reported alcohol-use and sexual risk-behaviors, while controlling for other known correlates of risk-taking such as impulsivity, sensation seeking, and peer norms among a group of high-risk African American female adolescents to evaluate whether this biological factor enhances our understanding of patterns of risk in this vulnerable group.

Socioeconomic-related risk and sexually transmitted infection among African-American adolescent females.

PURPOSE: Virtually no studies have examined the potential role that chronic stress, particularly the stress associated with socioeconomic status (SES) strain, may play on sexually transmitted infection (STI) risk. This study examined the association between SES-related risk at baseline to STI acquisition and reinfection over 36 months of follow-up. METHODS: Six hundred twenty-seven African-American female adolescents, ages 14-20 years, recruited from sexual health clinics in Atlanta, GA, participated in a randomized controlled HIV prevention trial and returned for at least one follow-up assessment. Following baseline assessment, six waves of data collection occurred prospectively over 36 months. Chronic SES-related risk was assessed as a sum of yes-no exposure to seven risk indicators. Laboratory-confirmed tests for Chlamydia trachomatis and Neisseria gonorrhoeae were performed at each follow-up. RESULTS: In multivariable regression analysis, SES-related risk significantly predicted STI acquisition over 36 months (adjusted odds ratio = 1.22) and STI reinfection (adjusted odds ratio = 1.16) above and beyond other known correlates of STI. CONCLUSIONS: Findings demonstrate that SES-related risk was predictive of both STI acquisition and reinfection among young African-American females. They are consistent with propositions that some health disparities observed in adulthood may be linked to earlier chronically stress-inducing life experiences, particularly experiences associated with low SES conditions. Although various explanations exist for the observed connection between SES-related risk and subsequent STI acquisition and/or reinfection across 36 months of follow-up, these findings highlight the need for further research to elucidate the exact pathway(s) by which SES-related risk influences later STI acquisition to refine STI prevention interventions for this population.

Factors associated with sexual arousal, sexual sensation seeking and sexual satisfaction among female African American adolescents.

BACKGROUND: Sexuality-related constructs, such as sexual arousal, sexual sensation seeking (SSS) and sexual satisfaction, have been related to sexual behaviours that place one at risk of adverse consequences, such as sexually transmissible infections, HIV and unintended pregnancy. The biopsychosocial model posits an array of factors, ranging from social environmental factors to biological and psychological predispositions, that may be associated with these sexuality constructs in adolescents. METHODS: Female African Americans aged 14-20 years were recruited from reproductive health clinics for an HIV intervention. Baseline survey and follow-up DNA data (n=304) were used to assess biological, psychological and social environmental associations with the sexuality constructs of arousal, SSS and sexual satisfaction. RESULTS: Multivariate linear regression analysis revealed that a higher depressive symptom rating was associated with higher arousability, whereas short serotonin transporter gene allele(s) status was associated with lower arousability. Impulsivity and perceived peer norms supportive of unsafe sexual behaviours were associated with increased SSS, whereas short serotonin transporter gene allele(s) status was associated with lower SSS. Higher social support was associated with higher levels of sexual satisfaction, whereas short serotonin transporter gene allele(s) status was associated with lower satisfaction. The sexuality constructs were also significantly related to the number of sex partners, the frequency of vaginal sex and the number of unprotected vaginal sex acts in the past 6 months. CONCLUSIONS: The findings emphasise the importance of understanding biopsychosocial factors, including the role of serotonin as an indicator of natural variations in sexual inclination and behaviours, that influence sexuality constructs, which, in turn, are associated with sexual behaviours, to allow further refinement of sexual health clinical services and programs and promote the development of healthy sexuality.