Examination of ATM, BRCA1, and BRCA2 promoter methylation in patients with pancreatic cancer.

BACKGROUND: Pancreatic cancer is a lethal disease with a poor 5-year survival rate. Pathogenic germline variants in the coding regions of ATM, BRCA1, and BRCA2 are found in up to 4.8% of pancreatic cancer patients. Germline promoter methylation and gene silencing arising from a germline variant or through other mechanisms have been described as a cause of tumor suppressor gene inactivation. METHODS: We measured the level of promoter methylation of the ATM, BRCA1, and BRCA2 genes in peripheral blood lymphocytes from 655 patients with pancreatic cancer using real-time PCR. RESULTS: No evidence of germline promoter methylation of any of these genes was found. Promoter methylation levels were minimal with no patient having promoter methylation greater than 3.4%, 3.3%, and 7.6% for ATM, BRCA1 and BRCA2, respectively, well below levels found in patients who have inherited promoter methylation (∼50%). CONCLUSIONS: We found no evidence of germline promoter methylation for the pancreatic susceptibility genes ATM, BRCA1 and BRCA2 in patients with pancreatic cancer. This study reveals that constitutive germline methylation of promoter CpG islands is rare in pancreatic cancer.

A Novel Scoring System for Humane Endpoints in Mice with Cecal Ligation and Puncture-Induced Sepsis.

Animal-based research is essential to the study of sepsis pathophysiology, diagnostics, and therapeutics. However, animal models of sepsis are often associated with high mortality because of the difficulty in predicting imminent death based on premortem assessment of the animals. The use of validated visual scoring would allow researchers to systematically identify humane endpoints but visual approaches require high interobserver agreement for accurate results. The objective of this study was to establish a scoring system for mice undergoing cecal ligation and puncture (CLP)-induced sepsis based on 3 visual parameters: respiratory status, activity and response to stimulus (ASR), and eye appearance, with scores ranging from 0 to 3. In the first study, we evaluated interobserver agreement. Veterinary and investigative staff assessed 283 mice with CLP and had substantial to near-perfect agreement for all 3 parameters as evaluated using weighted Cohen κ statistic. The second study assessed the ability of the scoring system and temperature to predict death. The scoring system and subcutaneous transpond- ers were used to monitor C57BL/6J mice (n = 80, male and female) until death or for 7 days after CLP. Results showed that the scoring system discriminates between surviving (n = 26) and nonsurviving (n = 54) septic mice. The scoring system was accurate in predicting death, with an AUC of 0.8997. The sensitivity and specificity of the ASR parameter were 96% and 92%, respectively, and for the eye parameter were 94% and 73%. A sum of the ASR and eye scores that was 5 or more was also predictive of death. Temperature was a quantitative predictor, with sensitivity and specificity of 93% and 92%, respectively. This scoring system refines the CLP model by allowing identification of humane endpoints and avoidance of spontaneous death.

Dietary resveratrol increases mid-life fecundity of female Nothobranchius guentheri.

The decline of female reproductive function is an early phenotype of aging in humans, occurring only midway through the lifespan. Yet the number of women delaying pregnancy continues to rise in industrialized societies due to personal or socioeconomic circumstances, often resulting in subfertility or difficulty conceiving. There are few defined mechanisms associated with this etiology, and equally few effective therapies. To combat this problem, we used a novel emerging model, Nothobranchius guentheri, that recapitulates the age-associated spectrum of changes that adversely affect human fertility. We hypothesized that resveratrol (RSV), which activates SirT1 as an oxidative stress sensor and longevity assurance enzyme, would improve female fecundity in mid-life. RSV, a polyphenol found in grapes and red wine, has been touted as an anti-aging dietary supplement due to its ability to prolong both lifespan and health span. SirT1 is an NAD+ dependent histone deacetylase, whose activity is regulated by the nicotinamide to NAD+ salvage pathway, especially the rate-limiting enzyme NAMPT. We found that female N. guentheri fed 600µgRSV/g food into mid-life (~20weeks), beginning at sexual maturity, showed increased embryo production compared to those on Control diet. Furthermore, the RSV-fed fish had significantly increased NAMPT. This suggests that dietary RSV has a positive effect on female fertility, and that it may become an effective therapy to regulate sirtuin activity and combat reproductive senescence.

Mismatched concepts in a neonatal intensive care unit (NICU): further issues for computer decision support?

BACKGROUND: Common concepts and definitions are important for the effective practice of medicine. In an intensive care unit clear understanding of terminology and communication between different staff groups may be critical for optimal care. If computerised decision support tools are to be successfully deployed in these high intensity environments, all staff must understand the concepts and information that is to be portrayed. OBJECTIVE: To examine the similarity of language and concepts related to newborn infants and their care in staff groups with different experience. METHODS: An experimental study in a tertiary regional neonatal intensive care unit involved 32 staff in 4 groups with varying experience (junior and senior / nurses and doctors). A psychologist developed a lexicon of clinical actions and a second lexicon of possible patient descriptors applicable to newborn infants receiving all degrees of neonatal care by conducting interviews with staff on the unit. Card sorting experiments were performed on the terms in the action and descriptor lexicons, the staff being asked to group words that were related to the same or similar concepts. The card sort data were analysed using conventional cluster analysis to produce tree-diagrams or dendrograms and then by distance matrix analysis to give cumulative probability plots. RESULTS: Differences were shown in the way various classes of staff and staff with different experience mentally map clinical concepts. Clinical actions were grouped more randomly by nurses and by those with less experience with a polarisation between senior doctors and junior nurses. Descriptors were classed more definitively and similarly by junior and senior nurses and senior doctors but more randomly and quite differently by junior doctors. Thus there were differences seen between nurses and doctors with different roles within the unit and differences related to experience. CONCLUSIONS: Concepts are used differently by various staff groups in a neonatal unit: this may have an impact on the effectiveness of computerised decision aids unless it is taken into account during their development.

Role and experience determine decision support interface requirements in a neonatal intensive care environment.

The aim of this paper is to describe a novel approach to the analysis of data obtained from card-sorting experiments. These experiments were performed as a part of the initial phase of a project, called NEONATE. One of the aims of the project is to develop decision support tools for the neonatal intensive care environment. Physical card-sorts were performed using clinical "action" and patient "descriptor" words. Thirty-two staff (eight junior nurses, eight senior nurses, eight junior doctors, and eight senior doctors) participated in the actions card-sorts and the same number of staff participated in separate descriptors card-sorting experiments. To check for consistency, the card-sorts were replicated for nurses during the action card-sorts. The card-sort data were analysed using hierarchical cluster analysis to produce tree-diagrams or dendrograms. Differences were shown in the way various classes of staff with different levels of experience mentally map clinical concepts. Clinical actions were grouped more loosely by nurses and by those with less experience, with a polarisation between senior doctors and junior nurses. Descriptors were classed more definitively and similarly by nurses and senior doctors but in a less structured way and quite differently by junior doctors. This paper presents a summary of the differences in the card-sort data for the various staff categories. It is shown that concepts are used differently by various staff groups in a neonatal unit and that this may diminish the effectiveness of computerised decision aids unless it is explored during their development.