Stability of the human sperm DNA methylome to folic acid fortification and short-term supplementation.

STUDY QUESTION: Do short-term and long-term exposures to low-dose folic acid supplementation alter DNA methylation in sperm? SUMMARY ANSWER: No alterations in sperm DNA methylation patterns were found following the administration of low-dose folic acid supplements of 400 µg/day for 90 days (short-term exposure) or when pre-fortification of food with folic acid and post-fortification sperm samples (long-term exposure) were compared. WHAT IS KNOWN ALREADY: Excess dietary folate may be detrimental to health and DNA methylation profiles due to folate's role in one-carbon metabolism and the formation of S-adenosyl methionine, the universal methyl donor. DNA methylation patterns are established in developing male germ cells and have been suggested to be affected by high-dose (5 mg/day) folic acid supplementation. STUDY DESIGN, SIZE, DURATION: This is a control versus treatment study where genome-wide sperm DNA methylation patterns were examined prior to fortification of food (1996-1997) in men with no history of infertility at baseline and following 90-day exposure to placebo (n = 9) or supplement containing 400 µg folic acid/day (n = 10). Additionally, pre-fortification sperm DNA methylation profiles (n = 19) were compared with those of a group of post-fortification (post-2004) men (n = 8) who had been exposed for several years to dietary folic acid fortification. PARTICIPANTS/MATERIALS, SETTING, METHODS: Blood and seminal plasma folate levels were measured in participants before and following the 90-day treatment with placebo or supplement. Sperm DNA methylation was assessed using the whole-genome and genome-wide techniques, MassArray epityper, restriction landmark genomic scanning, methyl-CpG immunoprecipitation and Illumina HumanMethylation450 Bead Array. MAIN RESULTS AND THE ROLE OF CHANCE: Following treatment, supplemented individuals had significantly higher levels of blood and seminal plasma folates compared to placebo. Initial first-generation genome-wide analyses of sperm DNA methylation showed little evidence of changes when comparing pre- and post-treatment samples. With Illumina HumanMethylation450 BeadChip arrays, no significant changes were observed in individual probes following low-level supplementation; when compared with those of the post-fortification cohort, there were also few differences in methylation despite exposure to years of fortified foods. LARGE SCALE DATA: Illumina HumanMethylation450 BeadChip data from this study have been submitted to the NCBI Gene Expression Omnibus under the accession number GSE89781. LIMITATIONS, REASONS FOR CAUTION: This study was limited to the number of participants available in each cohort, in particular those who were not exposed to early (pre-1998) fortification of food with folic acid. While genome-wide DNA methylation was assessed with several techniques that targeted genic and CpG-rich regions, intergenic regions were less well interrogated. WIDER IMPLICATIONS OF THE FINDINGS: Overall, our findings provide evidence that short-term exposure to low-dose folic acid supplements of 400 µg/day, over a period of 3 months, a duration of time that might occur during infertility treatments, has no major impact on the sperm DNA methylome. STUDY FUNDING/COMPETING INTERESTS: This work was supported by a grant to J.M.T. from the Canadian Institutes of Health Research (CIHR: MOP-89944). The authors have no conflicts of interest to declare.

Temporal and geographic clustering of polyomavirus-associated olfactory tumors in 10 free-ranging raccoons (Procyon lotor).

Reports of primary nervous system tumors in wild raccoons are extremely rare. Olfactory tumors were diagnosed postmortem in 9 free-ranging raccoons from 4 contiguous counties in California and 1 raccoon from Oregon within a 26-month period between 2010 and 2012. We describe the geographic and temporal features of these 10 cases, including the laboratory diagnostic investigations and the neuropathologic, immunohistochemical, and ultrastructural characteristics of these tumors in the affected animals. All 9 raccoons from California were found within a localized geographic region of the San Francisco Bay Area (within a 44.13-km radius). The tight temporal and geographic clustering and consistent anatomic location in the olfactory system of tumor types not previously described in raccoons (malignant peripheral nerve sheath tumors and undifferentiated sarcomas) strongly suggest either a common cause or a precipitating factor leading to induction or potentiation of neuro-oncogenesis and so prompted an extensive diagnostic investigation to explore possible oncogenic infectious and/or toxic causes. By a consensus polymerase chain reaction strategy, a novel, recently reported polyomavirus called raccoon polyomavirus was identified in all 10 tumors but not in the normal brain tissue from the affected animals, suggesting that the virus might play a role in neuro-oncogenesis. In addition, expression of the viral protein T antigen was detected in all tumors containing the viral sequences. We discuss the potential role of raccoon polyomavirus as an oncogenic virus.

A focus group study of DES daughters: implications for health care providers.

A focus group study of women exposed to diethylstilbestrol (DES) in utero (DES daughters) was conducted to gain understanding about exposure to this drug from a patient perspective. Focus group participants reported that learning about their DES exposure was devastating; they experienced strains in their family relationships, emotional shock, a feeling that their health concerns were not appreciated by others and, to some degree, a sense of social isolation. Although many were aware of the need for special gynecological exams and high-risk prenatal care, they were frustrated by what they felt was a lack of reliable and clear information about the effects of DES exposure. Most expressed questions and anxiety about their health. Many found their communication with physicians about their DES exposure unsatisfying. They felt that physicians lacked information about the long-term health effects of DES exposure and as a result did not give them accurate information. Furthermore, they felt that physicians were dismissive of their concerns and often gave what they felt to be false reassurances. Consequently, the women developed an enduring distrust of the medical profession. The results of the study suggest implications for the delivery of health care to DES daughters.

Awake intubation of the adult trachea using the Bullard laryngoscope.

Awake intubation using the Bullard laryngoscope can be comfortably and easily performed in the adult. Five cases are presented in which tracheal intubation was performed under topical anaesthesia with light intravenous sedation. In each case, topical anaesthesia was performed by insertion of a Guedel oral airway, with lidocaine ointment applied to the inferior and posterior surfaces. In one case, Bullard intubation was successful where direct laryngoscopy and multiple attempts at bronchoscopic intubation by three different operators had failed. We conclude that the Bullard laryngoscope can be easily used in awake patients and may be a useful alternative where other methods for awake intubation have failed.

CATCH: process evaluation of environmental factors and programs.

The Child and Adolescent Trial for Cardiovascular Health (CATCH) is a multicenter trial designed to test the effectiveness of school and family-based cardiovascular health promotion for preadolescents. CATCH interventions target multiple cardiovascular health behaviors such as dietary intake of fat and sodium, physical activity, and tobacco use. Evaluation includes physiological, psychosocial, behavioral, and process measures. An important aspect of the process evaluation is the assessment of environmental factors and "secular events" in both intervention and control schools that may affect outcomes independently of the CATCH interventions. With such information, CATCH investigators are able to isolate the impact of the CATCH intervention from competing (non-CATCH) factors as well as "track" proximal (i.e., immediate and short term) changes related to the intervention that may in turn lead to "distal" (long-term) behavior change. The School Health Questionnaire, the major process evaluation tool for monitoring secular and environmental changes in the schools, is described in detail, and data describing tobacco-related environmental factors and secular events are presented.

Sociocultural factors associated with smoking behavior by Puerto Rican adolescents in Boston.

This study examined cigarette smoking among Puerto Rican adolescents in the Boston area in its sociocultural context by describing who uses cigarettes, and by identifying factors that encourage or discourage cigarette use. The study, conducted from 1986-87, used two approaches. The first was to survey 605 Puerto Rican households with adolescents 11-20 years old. Households were identified from a random sample of census blocks in five neighborhoods. In each household, interviews were completed with one adolescent and his or her female caretaker. The second approach was to conduct ethnographic interviews with 40 adolescents selected through a purposive sample of survey respondents from three of the study neighborhoods. Bivariate and multivariate statistical analyses were used to examine the relationships between smoking behavior and characteristics of the adolescents. The results of these analyses were examined in light of the ethnographic data. Overall, 13.7% reported smoking in the month preceding the interview. Smoking was most prevalent among males 17-20. Patterns of smoking were associated with teens' social networks. The Puerto Rican adolescents in this study were more likely to smoke when their friends and household members smoked, if they participated in recreational activities such as sports or unsupervised games, and if they were not in school. These effects varied according to the gender of the respondent.

Evaluation of two community-wide smoking cessation contests.

The Pawtucket (Rhode Island) Heart Health Program is designed to effect a community-wide change in heart disease morbidity and mortality by reducing smoking prevalence and other behavioral risks for this disease. The initial emphasis of the Pawtucket Heart Health Program was on the development of risk factor programs within specific churches, work sites, and other organizations in the community. At the end of 9 months of programming, however, only one organization had elected to begin a stop-smoking program. Given the community smoking prevalence of 43.4%, it was decided that larger-scale interventions would be required if a significant public health impact was to be realized. Therefore, a community-wide campaign was mounted to recruit as many participants as possible into the "Up in Smoke" cessation program. A lottery was attached to this program, with the contingency based on program attendance rather than cessation per se. One hundred three participants, including residents of neighboring communities, enrolled in three Up in Smoke lottery groups. At a 3-month follow-up, only 11 (7%) of smokers from the Up in Smoke lottery reported that they were not smoking (10% of those actually contacted). For a variety of reasons, the "Quit and Win" approach was later adopted by the Pawtucket Heart Health Program as the primary smoking intervention. One month after the end of the contest, 20% of those contacted reported not smoking. The percentage of quitters for the Up in Smoke program increased while the Quit and Win rate decreased over longer periods of follow-up. These and other data were compared with those of participants of a screening program conducted concurrently. Lotteries in general and face-to-face recruitment in large crowds were shown to be effective recruiting methods for large-scale smoking cessation programs.

Organizational and community approaches to community-wide prevention of heart disease: the first two years of the Pawtucket Heart Health Program.

The Pawtucket Heart Health program (PHHP) is a federally funded research and demonstration project for the primary prevention of coronary heart disease (CHD) in a community. This article presents a discussion of the first 26 months of this intervention, divided into its three phases. PHHP staff initially approached the intervention city through local organizations to accomplish risk-factor behavior change in the population. After 11 months, PHHP complemented its programs in organizations with activities open to all city residents, in order to accelerate participation by the population. Seven months into this phase, it was decided that community activities should be the major focus of the intervention approach to assure a level of participation adequate to make a measurable impact. The third has shown the greatest percentage of public participation, demonstrating the complementary nature of organization and community interventions and of the translation of social learning theory into principles for primary prevention in a community.

Community intervention to lower blood cholesterol: the "Know Your Cholesterol" campaign in Pawtucket, Rhode Island.

Sufficient data exist to support approaches to lowering cardiovascular disease risk which target population-wide reductions in elevated blood cholesterol levels. Within the context of a multifactorial cardiovascular disease prevention program, a 2-month cholesterol education campaign was launched to encourage citizens to "Know Your Cholesterol" by attending screening, counseling, and referral events (SCOREs) held throughout the community. These events featured assessment of dietary fat and cholesterol, rapid blood cholesterol analyses with a fingerstick procedure and the Kodak Ektachem DT60, and immediate dietary counseling by trained volunteers. A total of 1439 persons attended 39 SCOREs: nearly 60% were found to have blood cholesterol levels that exceeded recommended goal levels. At a 2-month follow-up of 1040 of these same individuals, 600 had lowered their blood cholesterol level by an average of 29.1 mg/dl. This experience suggests that similarly modeled campaigns may have a significant impact on the distribution of blood cholesterol levels in the entire U.S. population.

Laryngeal papillomatosis: anesthetic management.

We reviewed the anesthetic management of all children admitted with laryngeal papillomatosis during a six-year period. This included 147 endoscopies on 16 patients, most referred with advanced disease after initial care in other hospitals. Onset of the disease before age 2 was associated with the most severe upper airway involvement. Eight patients required tracheostomy at some time during treatment, and management of the shared airway during surgical manipulation was particularly difficult in these children. Inhalational induction with halothane was used rountinely, with surprisingly few instances of laryngospasm or upper airway obstruction. Topical lidocaine spray and orotracheal intubation were preferred; insufflation technics were frequently abandoned when attempted. Extubation under deep anesthesia after clearing the glottis of secretions was usually followed by smooth emergence and minimal airway difficulties. There was only one instance of postoperative stridor suggesting croup. A standard anesthetic approach can be recommended, on the basis of the experience in this series, but possible variations in technic are also discussed for particular clinical applications.