Genome Editing Technology, CRISPR Cas-9, Provides a Potential Future for Xenotransplantation: Can Pigs Become Our New Organ Donors?

Organ shortage has been a significant issue in United States for many decades. There were over 116,000 people on the transplant waiting list as of August 2017, with a new person added every 10 minutes. Animals have been considered as potential organ donors for humans, with the pig being the most ideal candidate because of its excellent breeding profile, low maintenance costs, large litters, rapid growth and organ similarity. A notable challenge that comes with porcine organs are endogenous retroviruses that establish themselves in the DNA of offspring in utero, thereby allowing for no time to prevent infection and integration. Porcine endogenous retroviruses (PERVs) have the ability to infect human cells and therefore must be removed or inactivated before considering transplantation. Genome editing technology, CRISPR-cas9, has been recently successful in targeting and inactivating PERVs in porcine fetal cell lines. Using PERVinactivated cells, reproductive technologies have allowed for fertilization, implantation and the successful birth of 37 PERV-inactivated piglets from 17 sows. At this time, 15 piglets remain alive with the oldest piglet being 4 months of age. The success of this technology takes us one step closer to finding potential organ donors outside of our own species and instilling hope back into those who may need an organ transplant in the future

Correlation functions quantify super-resolution images and estimate apparent clustering due to over-counting

We present an analytical method to quantify clustering in super-resolution localization images of static surfaces in two dimensions. The method also describes how over-counting of labeled molecules contributes to apparent self-clustering and how the effective lateral resolution of an image can be determined. This treatment applies to clustering of proteins and lipids in membranes, where there is significant interest in using super-resolution localization techniques to probe membrane heterogeneity. When images are quantified using pair correlation functions, the magnitude of apparent clustering due to over-counting will vary inversely with the surface density of labeled molecules and does not depend on the number of times an average molecule is counted. Over-counting does not yield apparent co-clustering in double label experiments when pair cross-correlation functions are measured. We apply our analytical method to quantify the distribution of the IgE receptor (Fc{\epsilon}RI) on the plasma membranes of chemically fixed RBL-2H3 mast cells from images acquired using stochastic optical reconstruction microscopy (STORM) and scanning electron microscopy (SEM). We find that apparent clustering of labeled IgE bound to Fc{\epsilon}RI detected with both methods arises from over-counting of individual complexes. Thus our results indicate that these receptors are randomly distributed within the resolution and sensitivity limits of these experiments.Comment: 22 pages, 5 figure

Small business owners' health and safety intentions: A cross-sectional survey

BACKGROUND: Little is known about the variables underlying small business owners' behavioural intentions toward workplace health and safety. This project explores the relationship between three mediating variables (Attitude Toward Safety, Subjective Norm and Perceived Behavioural Control) and owners' Intentions Toward Safety, following the Theory of Planned Behaviour. We also investigate the role of beliefs underlying each mediating variable. METHODS: Seven hundred businesses (5–50 employees) were randomly selected from 4084 eligible companies in a manufacturing business database (SIC codes 24 to 39). The 348 respondents are on average 51 yrs of age, 86% male, 96% white and have 2 to 4 years of post-secondary school. RESULTS: All three mediator variables are significantly correlated with Intentions Toward Safety; Attitude Toward Safety shows the strongest correlation, which is confirmed by path analysis. Owners with higher attitudes toward safety have a higher probability of believing that improving workplace health and safety will make employees' healthier and happier, show that they care, increase employee productivity, lower workers' compensation costs, increase product quality and lower costs. CONCLUSION: These results suggest that interventions aimed at increasing owners' health and safety intentions (and thus, behaviours) should focus on demonstrating positive employee health and product quality outcomes

Evolution of sex-specific pace-of-life syndromes: genetic architecture and physiological mechanisms

Sex differences in life history, physiology, and behavior are nearly ubiquitous across taxa, owing to sex-specific selection that arises from different reproductive strategies of the sexes. The pace-of-life syndrome (POLS) hypothesis predicts that most variation in such traits among individuals, populations, and species falls along a slow-fast pace-of-life continuum. As a result of their different reproductive roles and environment, the sexes also commonly differ in pace-of-life, with important consequences for the evolution of POLS. Here, we outline mechanisms for how males and females can evolve differences in POLS traits and in how such traits can covary differently despite constraints resulting from a shared genome. We review the current knowledge of the genetic basis of POLS traits and suggest candidate genes and pathways for future studies. Pleiotropic effects may govern many of the genetic correlations, but little is still known about the mechanisms involved in trade-offs between current and future reproduction and their integration with behavioral variation. We highlight the importance of metabolic and hormonal pathways in mediating sex differences in POLS traits; however, there is still a shortage of studies that test for sex specificity in molecular effects and their evolutionary causes. Considering whether and how sexual dimorphism evolves in POLS traits provides a more holistic framework to understand how behavioral variation is integrated with life histories and physiology, and we call for studies that focus on examining the sex-specific genetic architecture of this integration

Intraoperative Syndesmotic Instability Test

Category: Ankle, Trauma Introduction/Purpose: Precise diagnosis of distal tibiofibular syndesmotic injury is challenging and a gold standard diagnostic test has still not been established. Tibiofibular clear space identified on radiographic imaging is considered the most reliable indicator of the injury. The Cotton test is the most widely used intraoperative technique to evaluate the syndesmotic integrity although it has its limitations. We advocate for a novel intra operative test using a 3.5 mm cortical tap. Methods: Tibiofibular clear space was assessed in nine cadaveric specimens using three sequential fluoroscopic images. The first image was taken prior to the application of the tap test representing the intact and non-stressed state. Then, a 2.5 mm hole was drilled distally on the lateral fibula, and a 3.5 mm cortical tap was then threaded in the hole. The tap test involved gradually advancing the blunt tip against the lateral tibia, providing a tibiofibular separation force (intact, stressed). This same stress was then applied after all syndesmotic ligaments were released (injured, stressed). Measurements were compared by one-way ANOVA and paired Student’s t-test. Intra and inter-observer agreements were evaluated by intraclass correlation coefficient (ICC). P-values 6 mm as diagnostic for syndesmotic instability, the tap test demonstrated a 96.3% sensitivity and specificity, a 96.3% PPV and NPV and a 96.3% accuracy in diagnosing syndesmotic instability. Conclusion: Our cadaveric study showed that this novel syndesmotic instability test using a 3.5 mm blunt cortical tap is a simple, accurate and reliable technique able to demonstrate significant differences in the tibiofibular clear space when injury was present. It could represent a more controlled and stable low-cost alternative to the most used Cotton test