The effects of elevated potassium, acidosis, reduced oxygen levels, and temperature on the functional properties of isolated myocardium from three elasmobranch fishes: clearnose skate (Rostroraja eglanteria), smooth dogfish (Mustelus canis), and sandbar shark (Carcharhinus plumbeus).

Elevated plasma potassium levels (hyperkalemia), reduced plasma pH (acidosis), reduced blood oxygen content, and elevated temperatures are associated with species-specific rates of at-vessel and post-release mortality in elasmobranch fishes. The mechanism linking these physiological disturbances to mortality remains undetermined however, and we hypothesize that the proximate cause is reduced myocardial function. We measured changes in the functional properties of isolated ventricular myocardial strips from clearnose skate (Rostroraja eglanteria), smooth dogfish (Mustelus canis), and sandbar shark (Carcharhinus plumbeus) when subjected to the following stressors (both in isolation and in combination): hyperkalemia (7.4 mM K+), acidosis (from 7.9 to 7.1), and reduced oxygen (to 31% O2 saturation) applied at temperatures 5 °C above and below holding temperatures. We selected these species based on phylogenetic distance, diverse routine activity levels, and their tolerance to capture and transport. Stressors had a few significant species-specific detrimental impacts on myocardial function (e.g., a 33-45% decrease in net force under acidosis + low O2). Net force production of myocardial strips from clearnose skate and smooth dogfish approximately doubled following exposure to isoproterenol, demonstrating that these species possess beta-adrenergic receptors and that their stimulation could provide a mechanism for preservation of cardiac function during stress. Our results suggest that disruption of physiological homeostasis associated with capture may fatally impair cardiac function in some elasmobranch species, although research with more severe stressors is needed.

Connecting post-release mortality to the physiological stress response of large coastal sharks in a commercial longline fishery.

Bycatch mortality is a major factor contributing to shark population declines. Post-release mortality (PRM) is particularly difficult to quantify, limiting the accuracy of stock assessments. We paired blood-stress physiology with animal-borne accelerometers to quantify PRM rates of sharks caught in a commercial bottom longline fishery. Blood was sampled from the same individuals that were tagged, providing direct correlation between stress physiology and animal fate for sandbar (Carcharhinus plumbeus, N = 130), blacktip (C. limbatus, N = 105), tiger (Galeocerdo cuvier, N = 52), spinner (C. brevipinna, N = 14), and bull sharks (C. leucas, N = 14). PRM rates ranged from 2% and 3% PRM in tiger and sandbar sharks to 42% and 71% PRM in blacktip and spinner sharks, respectively. Decision trees based on blood values predicted mortality with >67% accuracy in blacktip and spinner sharks, and >99% accuracy in sandbar sharks. Ninety percent of PRM occurred within 5 h after release and 59% within 2 h. Blood physiology indicated that PRM was primarily associated with acidosis and increases in plasma potassium levels. Total fishing mortality reached 62% for blacktip and 89% for spinner sharks, which may be under-estimates given that some soak times were shortened to focus on PRM. Our findings suggest that no-take regulations may be beneficial for sandbar, tiger, and bull sharks, but less effective for more susceptible species such as blacktip and spinner sharks.

Communicating effectively about donation: an educational intervention to increase consent to donation.

CONTEXT: Families' refusal to consent to solid organ donation is a major contributor to the organ deficit in the United States. Previous research has identified organ procurement coordinators as best able to obtain consent from families; however, few studies have examined the effects of coordinator training programs on consent rates. OBJECTIVE: To test the effects of the Communicating Effectively About Donation intervention on the rate of family consent to solid organ donation. DESIGN: A nonrandomized repeated measures design. SETTING AND PARTICIPANTS: Participants included 17 hospitals, 502 donor-eligible patients and their families, and 22 coordinators from an organ procurement organization in Ohio. INTERVENTION: Coordinators were given in-service training on the use of effective relational and affective communication techniques through a day-long interactive workshop and simulated donation scenarios. MAIN OUTCOME MEASURES: Families' final donation decision and coordinators' donation-related behaviors. RESULTS: Training of coordinators was associated with increases in coordinators' comfort speaking with patients' families about donation and answering donation-related questions, in the amount of time coordinators spent discussing donation with family members, and in the number of donation-related topics discussed with families. Consent rates increased from 46.3% to 55.5% after the intervention. CONCLUSIONS: The results suggest that improving coordinators' communication skills may be a fruitful avenue for increasing the rate of family consent to donation; however, a more definitive test of the training is needed to confirm the intervention's effectiveness.

Modulating target site selection during human immunodeficiency virus DNA integration in vitro with an engineered tethering factor.

The mechanisms controlling retroviral integration have been the topic of intense interest, in part because of adverse clinical events that occurred during retrovirus-mediated human gene therapy. Here we investigate the use of artificial tethering interactions to constrain retroviral integration site selection in an in vitro model. During normal infection, HIV DNA integration is favored in active cellular transcription units. One component of the targeting mechanism is the cellular LEDGF/p75 protein. LEDGF/p75 binds tightly to HIV integrase (IN) protein, and depletion of LEDGF/p75 from target cells results in reduced integration in transcription units, suggesting integration targeting by a tethering mechanism. We constructed and analyzed fusions of LEDGF/p75 or its IN-binding domain (IBD) to the DNA-binding domain of phage lambda repressor protein (lambdaR). In the presence of the lambdaR-LEDGF/p75 fusions, increased strand transfer by IN was seen in target DNA near lambdaR-binding sites in vitro . These data support the idea that a direct interaction between LEDGF/p75 and IN can mediate targeting via a tethering mechanism, and provide proof of concept for the idea that protein-protein interactions might be engineered to constrain integration site selection during human gene therapy.

The rapid assessment of hospital procurement barriers in donation: assessing hospitals for change.

Assessment of the hospital environment is integral to improving the organ donation process in healthcare organizations. This paper introduces a novel approach to the assessment and improvement of donation processes: the Rapid Assessment of hospital Procurement barriers in Donation (RAPiD). The RAPiD is a qualitative needs assessment tool for identifying barriers to donor identification and referral, and family requests for donation. Improving the donation process has become a national priority and can potentially save or improve the lives of thousands of Americans each year. The RAPID yields a rich description of the hospital environment that is readily translated into action-oriented recommendations for change.

Comparing college students' value-, outcome-, and impression-relevant involvement in health-related issues.

The purpose of this study was to examine the role of receiver involvement in the context of health communication. Students (N = 277) completed Cho and Boster's (2005) measures of value-, outcome-, and impression-relevant involvement across 6 health behaviors, including cigarette smoking, organ and tissue donation, sunscreen use, alcohol use, sexually transmitted disease testing, and nutrition. Confirmatory factor analyses across all 6 health topics provided evidence of the 3-factor structure conceptualized by Johnson and Eagly (1989) and measured by Cho and Boster (2005). When health behaviors were regressed onto value-, outcome-, and impression-relevant involvement, outcome- and value-involvement, generally speaking, emerged as significant predictors. Results and implications of considering health campaign audience members' levels of involvement are discussed in the domain of preventive medicine.

Role of PSIP1/LEDGF/p75 in lentiviral infectivity and integration targeting.

BACKGROUND: To replicate, lentiviruses such as HIV must integrate DNA copies of their RNA genomes into host cell chromosomes. Lentiviral integration is favored in active transcription units, which allows efficient viral gene expression after integration, but the mechanisms directing integration targeting are incompletely understood. A cellular protein, PSIP1/LEDGF/p75, binds tightly to the lentiviral-encoded integrase protein (IN), and has been reported to be important for HIV infectivity and integration targeting. METHODOLOGY: Here we report studies of lentiviral integration targeting in 1) human cells with intensified RNAi knockdowns of PSIP1/LEDGF/p75, and 2) murine cells with homozygous gene trap mutations in the PSIP1/LEDGF/p75 locus. Infections with vectors derived from equine infections anemia virus (EIAV) and HIV were compared. Integration acceptor sites were analyzed by DNA bar coding and pyrosequencing. CONCLUSIONS/SIGNIFICANCE: In both PSIP1/LEDGF/p75-depleted cell lines, reductions were seen in lentiviral infectivity compared to controls. For the human cells, integration was reduced in transcription units in the knockdowns, and this reduction was greater than in our previous studies of human cells less completely depleted for PSIP1/LEDGF/p75. For the homozygous mutant mouse cells, similar reductions in integration in transcription units were seen, paralleling a previous study of a different mutant mouse line. Integration did not become random, however-integration in transcription units in both cell types was still favored, though to a reduced degree. New trends also appeared, including favored integration near CpG islands. In addition, we carried out a bioinformatic study of 15 HIV integration site data sets in different cell types, which showed that the frequency of integration in transcription units was correlated with the cell-type specific levels of PSIP1/LEDGF/p75 expression.