From bacteria to humans: the CRISPR/Cas9 editing system, a novel therapeutic

CRISPR (clustered regulatory interspaced short palindromic repeats) is a bacterial adaptive immune system, which can target specific sequences of foreign nucleic acids. The system is made up of unique spacer sequences flanked by short, repetitive, palindromic sequences, and sequences encoding Cas (CRISPR associated) proteins. The CRISPR-Cas9 system has been utilized from S. pyrogenes for use in eukaryotes for gene editing. Many years were spent uncovering the components of the system and its mechanism. Now the RNA-guided endonuclease abilities of the system are utilized to induce double-strand breaks in targeted DNA sequences. Through the advent of double-strand breaks researchers can now induce mutations, knock out genes through non-homologous end joining, and knock-in DNA sequences through homologous directed repair. Additionally, the system has been utilized in gene regulation to activate or repress protein expression. Studies have shown that CRISPR-Cas9 as a gene-editing tool is highly efficient and specific. The system only requires a Cas9 protein and guideRNA to function and can alter multiple genes with the addition of different guideRNA. Most scientists have reported minimal off-target mutations caused by Cas9, and many strategies have been initiated to reduce off-target effects even further. Already, the system has been shown to have therapeutic applications both in vitro in human cells and in vivo in animals (including non-human primates). Future applications in therapeutics and gene-editing have the potential to change research and medicine

“Catch 22”: biosecurity awareness, interpretation and practice amongst poultry catchers

Campylobacter contamination of chicken on sale in the UK remains at high levels and has a substantial public health impact. This has prompted the application of many interventions in the supply chain, including enhanced biosecurity measures on-farm. Catching and thinning are acknowledged as threats to the maintenance of good biosecurity, yet the people employed to undertake this critical work (i.e. ‘catchers’) are a rarely studied group. This study uses a mixed methods approach to investigate catchers’ (n = 53) understanding of the biosecurity threats posed by the catching and thinning, and the barriers to good biosecurity practice. It interrogated the role of training in both the awareness and practice of good biosecurity. Awareness of lapses in biosecurity was assessed using a Watch-&-Click hazard awareness survey (n = 53). Qualitative interviews (n = 49 catchers, 5 farm managers) explored the understanding, experience and practice of catching and biosecurity. All of the catchers who took part in the Watch-&-Click study identified at least one of the biosecurity threats with 40% detecting all of the hazards. Those who had undergone training were significantly more likely to identify specific biosecurity threats and have a higher awareness score overall (48% compared to 9%, p = 0.03). Crucially, the individual and group interviews revealed the tensions between the high levels of biosecurity awareness evident from the survey and the reality of the routine practice of catching and thinning. Time pressures and a lack of equipment rather than a lack of knowledge appear a more fundamental cause of catcher-related biosecurity lapses. Our results reveal that catchers find themselves in a ‘catch-22′ situation in which mutually conflicting circumstances prevent simultaneous completion of their job and compliance with biosecurity standards

Differential function and maturation of human stem cell-derived islets after transplantation

Insulin-producing stem cell-derived islets (SC-islets) provide a virtually unlimited cell source for diabetes cell replacement therapy. While SC-islets are less functional when first differentiated in vitro compared to isolated cadaveric islets, transplantation into mice has been shown to increase their maturation. To understand the effects of transplantation on maturation and function of SC-islets, we examined the effects of cell dose, transplantation strategy, and diabetic state in immunocompromised mice. Transplantation of 2 and 5, but not 0.75 million SC-islet cells underneath the kidney capsule successfully reversed diabetes in mice with pre-existing diabetes. SQ and intramuscular injections failed to reverse diabetes at all doses and had undetectable expression of maturation markers, such as MAFA and FAM159B. Furthermore, SC-islets had similar function and maturation marker expression regardless of diabetic state. Our results illustrate that transplantation parameters are linked to SC-islet function and maturation, providing ideal mouse models for preclinical diabetes SC therapy research

Barrow and Gilt Vocalizations during a Human Approach Test

The objective of this study was to investigate differences between barrow and gilt vocalizations during a fear test. Twenty barrows and 20 gilts were tested over two consecutive weeks between 1300 and 1900 hours using a human approach test (HAT). Throughout the test, vocalizations were recorded. Gilts expressed a greater number of low calls compared to barrows; however, barrows expressed a greater number of high calls compared to gilts. Further research should be done to better understand vocalization differences between barrows and gilts during a HAT

Association of Vocalizations and Swine Behavior during a Human Approach Test

The objective of this study was to determine if the duration and total number of pig vocalizations when divided into low and high call categories was related to pig behaviors during a fear test. Twenty barrows and 20 gilts were tested over two consecutive weeks between 1300 and 1900 hours using a human approach test (HAT). Throughout the test, vocalizations and behavior were recorded. These results suggest that while high calls are typically the primary measure of stress vocalizations, low calls are also meaningful measures during the stressor of HAT

Identification of unique cell type responses in pancreatic islets to stress

Diabetes involves the death or dysfunction of pancreatic β-cells. Analysis of bulk sequencing from human samples and studies using in vitro and in vivo models suggest that endoplasmic reticulum and inflammatory signaling play an important role in diabetes progression. To better characterize cell type-specific stress response, we perform multiplexed single-cell RNA sequencing to define the transcriptional signature of primary human islet cells exposed to endoplasmic reticulum and inflammatory stress. Through comprehensive pair-wise analysis of stress responses across pancreatic endocrine and exocrine cell types, we define changes in gene expression for each cell type under different diabetes-associated stressors. We find that β-, α-, and ductal cells have the greatest transcriptional response. We utilize stem cell-derived islets to study islet health through the candidate gene CIB1, which was upregulated under stress in primary human islets. Our findings provide insights into cell type-specific responses to diabetes-associated stress and establish a resource to identify targets for diabetes therapeutics

Single-nucleus multi-omics of human stem cell-derived islets identifies deficiencies in lineage specification

Insulin-producing β cells created from human pluripotent stem cells have potential as a therapy for insulin-dependent diabetes, but human pluripotent stem cell-derived islets (SC-islets) still differ from their in vivo counterparts. To better understand the state of cell types within SC-islets and identify lineage specification deficiencies, we used single-nucleus multi-omic sequencing to analyse chromatin accessibility and transcriptional profiles of SC-islets and primary human islets. Here we provide an analysis that enabled the derivation of gene lists and activity for identifying each SC-islet cell type compared with primary islets. Within SC-islets, we found that the difference between β cells and awry enterochromaffin-like cells is a gradient of cell states rather than a stark difference in identity. Furthermore, transplantation of SC-islets in vivo improved cellular identities overtime, while long-term in vitro culture did not. Collectively, our results highlight the importance of chromatin and transcriptional landscapes during islet cell specification and maturation

Public safety personnel feedback from a remote trial of Goal Management Training for post-traumatic stress during Covid-19

Purpose: This paper explores participants’ perspectives on the acceptability, utility, and perceived therapeutic effects of a virtual group cognitive remediation program, Goal Management Training (GMT)™, during the COVID-19 pandemic. The advantages and drawbacks of these groups are considered as part of an online research study protocol exploring cognitive remediation among first responders (police, firefighters, paramedics, emergency dispatchers, corrections and parole officers, and nurses) who have been impacted by trauma. Methods: We qualitatively examined the results of an anonymous participant feedback survey collected from 20 first responders who took part in the first round of our online therapy groups. A thematic analysis approach was taken to highlight key themes and recommendations. Results: Survey results indicated that participants found our online protocol effective in terms of group facilitation, the utility of online platforms, and perceived therapeutic effects. Further, some participants preferred participating online versus attending in-person groups. Conclusion: This early data suggests that providing virtual options for research and treatment among trauma-impacted public safety personnel may increase accessibility and overall participation among this population

Restaurant cooking trends and increased risk for Campylobacter infection

In the United Kingdom, outbreaks of Campylobacter infection are increasingly attributed to undercooked chicken livers, yet many recipes, including those of top chefs, advocate short cooking times and serving livers pink. During 2015, we studied preferences of chefs and the public in the United Kingdom and investigated the link between liver rareness and survival of Campylobacter. We used photographs to assess chefs’ ability to identify chicken livers meeting safe cooking guidelines. To investigate the microbiological safety of livers chefs preferred to serve, we modeled Campylobacter survival in infected chicken livers cooked to various temperatures. Most chefs correctly identified safely cooked livers but overestimated the public’s preference for rareness and thus preferred to serve them more rare. We estimated that 19%-52% of livers served commercially in the United Kingdom fail to reach 70°C and that predicted Campylobacter survival rates are 48%-98%. These findings indicate that cooking trends are linked to increasing Campylobacter infections

Estimating the prevalence of food risk increasing behaviours in UK kitchens

© 2017 Jones et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Foodborne disease poses a serious threat to public health. In the UK, half a million cases are linked to known pathogens and more than half of all outbreaks are associated with catering establishments. The UK Food Standards Agency (FSA) has initiated the UK Food Hygiene Rating Scheme in which commercial food establishments are inspected and scored with the results made public. In this study we investigate the prevalence of food risk increasing behaviours among chefs, catering students and the public. Given the incentive for respondents to misreport when asked about illegal or illicit behaviours we employed a Randomised Response Technique designed to elicit more accurate prevalence rates of such behaviours. We found 14% of the public not always hand-washing immediately after handling raw meat, poultry or fish; 32% of chefs and catering students had worked within 48 hours of suffering from diarrhoea or vomiting. 22% of the public admitted having served meat “on the turn” and 33% of chefs and catering students admitted working in kitchens where such meat was served; 12% of the public and 16% of chefs and catering students admitted having served chicken at a barbeque when not totally sure it was fully cooked. Chefs in fine-dining establishment were less likely to wash their hands after handling meat and fish and those who worked in award winning restaurants were more likely to have returned to work within 48 hours of suffering from diarrhoea and vomiting. We found no correlation between the price of a meal in an establishment, nor its Food Hygiene Rating Score, and the likelihood of any of the food malpractices occurring