Bugs, Bumps, and Bacteria: The Role of Virtual Collaborative Learning Network in Pediatric Dermatology.

Patients in rural and underserved areas face significant barriers in accessing specialty care due to unavailability of services, geographic isolation, travel burden, and other cultural and socioeconomic factors.1 Pediatric dermatology is among the top three subspecialties that provides routine care for pediatric patients, however, shortage and maldistribution of pediatric dermatologists have remained a major hurdle for those living in remote and isolated areas.2 Pediatric dermatologists cluster in urban areas with high-patient volume and estimated wait times for new patients that often exceed 13 weeks, making access one of the major drivers of inequity for rural patients.2-4.

Gene Correction Recovers Phagocytosis in Retinal Pigment Epithelium Derived from Retinitis Pigmentosa-Human-Induced Pluripotent Stem Cells.

Hereditary retinal dystrophies (HRD) represent a significant cause of blindness, affecting mostly retinal pigment epithelium (RPE) and photoreceptors (PRs), and currently suffer from a lack of effective treatments. Highly specialized RPE and PR cells interact mutually in the functional retina, therefore primary HRD affecting one cell type leading to a secondary HRD in the other cells. Phagocytosis is one of the primary functions of the RPE and studies have discovered that mutations in the phagocytosis-associated gene Mer tyrosine kinase receptor (MERTK) lead to primary RPE dystrophy. Treatment strategies for this rare disease include the replacement of diseased RPE with healthy autologous RPE to prevent PR degeneration. The generation and directed differentiation of patient-derived human-induced pluripotent stem cells (hiPSCs) may provide a means to generate autologous therapeutically-relevant adult cells, including RPE and PR. However, the continued presence of the MERTK gene mutation in patient-derived hiPSCs represents a significant drawback. Recently, we reported the generation of a hiPSC model of MERTK-associated Retinitis Pigmentosa (RP) that recapitulates disease phenotype and the subsequent creation of gene-corrected RP-hiPSCs using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9. In this study, we differentiated gene-corrected RP-hiPSCs into RPE and found that these cells had recovered both wild-type MERTK protein expression and the lost phagocytosis of fluorescently-labeled photoreceptor outer segments observed in uncorrected RP-hiPSC-RPE. These findings provide proof-of-principle for the utility of gene-corrected hiPSCs as an unlimited cell source for personalized cell therapy of rare vision disorders.

Unraveling the Developmental Roadmap toward Human Brown Adipose Tissue.

Increasing brown adipose tissue (BAT) mass and activation is a therapeutic strategy to treat obesity and complications. Obese and diabetic patients possess low amounts of BAT, so an efficient way to expand their mass is necessary. There is limited knowledge about how human BAT develops, differentiates, and is optimally activated. Accessing human BAT is challenging, given its low volume and anatomical dispersion. These constraints make detailed BAT-related developmental and functional mechanistic studies in humans virtually impossible. We have developed and characterized functionally and molecularly a new chemically defined protocol for the differentiation of human pluripotent stem cells (hPSCs) into brown adipocytes (BAs) that overcomes current limitations. This protocol recapitulates step by step the physiological developmental path of human BAT. The BAs obtained express BA and thermogenic markers, are insulin sensitive, and responsive to ß-adrenergic stimuli. This new protocol is scalable, enabling the study of human BAs at early stages of development.

Risk-based early detection system of African Swine Fever using mortality thresholds.

African swine fever (ASF) is an infectious disease of swine causing major losses in the swine industry worldwide. Early detection of ASF is challenging because of the wide range of non-specific clinical signs produced and its relatively low contagiousness. Monitoring pig mortality is a promising approach for early detection of ASF, but such approach has been associated with delay in disease detection in large pig farms. The purpose of this study was to compare the effectiveness and suitability of early detection strategies for ASF in large commercial pig farms using mortality monitoring at the pen, room or barn level. The within-barn spread of the disease was modelled including the non-homogeneous probabilities of transmission within pens, between pens and between rooms. The performances of early detection surveillance based on mortality thresholds established for different epidemiological units were compared in terms of sensitivity, time to detection and number of false alarms per year. A barn with a capacity of 3,200 pigs divided into 8 rooms with 10 pens each containing 40 pigs per pen was used as an example. Our results show that using room- or pen-based mortality thresholds provided a time to detection of 8 days post-disease introduction. Similar detection performances could be achieved with barn-level mortality threshold but at the cost of an increased number of pigs to be tested each year. The different scenarios tested also show that barn characteristics such as baseline mortality rate and pen size had a limited impact on the pen-level mortality thresholds required for disease early detection. These results offer strong support for using mortality data for early detection of ASF not only in small pig herds but also in large commercial barns. Furthermore, the mortality thresholds defined in this study might be relevant to a wide range of pig production sites.

The association between vitamin D deficiency and hospitalization outcomes in pediatric patients with sickle cell disease.

BACKGROUND: While there is a known association between low vitamin D levels and increased chronic pain in patients with Sickle Cell Disease (SCD), there are no reported studies evaluating the relationship of vitamin D levels and hospitalization outcomes in this population. The aim of this study was to assess this relationship with hospitalization outcomes defined as the number of emergency room (ER) visits, hospital admissions for pain crisis, and length of hospital stay. DESIGN: A retrospective chart review of all pediatric patients with SCD (1-21 years old) was performed from January 2015 to January 2016 in an urban-based hospital setting (n = 134). Those with at least one reported Vitamin D level who maintained follow up during the time studied were enrolled (n = 90). Patient hospitalizations rates were compared between vitamin D deficiency (<20 ng/ml) and sufficiency (>20 ng/ml). RESULTS: Patients with both SCD and vitamin D deficiency were more likely to have at least one Emergency Room visit (p < 0.01), at least one admission for pain crisis (p < 0.01), and a longer length of admission (p < 0.0001) when compared to patients with SCD and sufficient vitamin D levels. CONCLUSION: There is a significant association between vitamin D levels of <20 ng/ml and the number of ER visits, hospital admissions for pain crisis, and length of stay in patients with SCD. Further research is required to assess if correcting vitamin D levels may improve hospitalization outcomes in this population.

Delaying the First Newborn Bath and Exclusive Breastfeeding.

PURPOSE: The purpose of this study was to determine if changing the timing of the initial newborn bath would have an impact on exclusive breastfeeding during hospitalization. The first newborn bath had been routinely done within 2 hours of age; practice was changed to the first bath being delayed until at least 12 hours of age. A secondary purpose was to examine the nurses' perceived benefits and challenges to such a change. STUDY DESIGN AND METHODS: Through a retrospective design, exclusive breastfeeding rates among mother-infant couplets prepractice change (cohort A) were compared with two postchange cohorts (B and C). Cohorts B and C were from the first 5 months and second 5 months, respectively after the practice change. Demographic information, birth type, bath timing, and feeding data were collected. Comparative statistics were applied to the three cohorts to examine differences in exclusive breastfeeding rates. Postpartum nurses were asked two open-ended questions on concerns and benefits of this change via an anonymous survey. Content analysis was completed on responses. RESULTS: There were 1,463 mother-infant couplets included in three cohorts (A: n = 564; B: n = 468; C: n = 431). There were no significant increases in the exclusive breastfeeding rates (baseline 74.1%) in both the first postimplementation delayed bath cohort (70.1%, p = .207) and the second "sustainability" cohort (79.4%, p = .060). Fifteen of the 60 postpartum nurses completed the survey, for a response rate of 25%. Themes generated from survey responses included concerns (infection control, work distribution), as well as benefits (perceived breastfeeding success, decreased workload) with delaying the first newborn bath. CLINICAL IMPLICATIONS: Delaying the first newborn bath may be one factor that can influence exclusive breastfeeding rates during postpartum hospitalization. Results have been mixed based on recent literature. In our study, the exclusive breastfeeding rate was already above average, as would be expected in a Baby-Friendly designated hospital and may be a reason we did not see a significant change in the rate among mother-infant couplets in our study. Randomized trials are needed for a rigorous evaluation of timing of the newborn bath and possible link to exclusive breastfeeding in the hospital and beyond.

Editing the Genome of Human Induced Pluripotent Stem Cells Using CRISPR/Cas9 Ribonucleoprotein Complexes.

Genome editing using the CRISPR/Cas9 system has rapidly established itself as an essential tool in the genetic manipulation of many organisms, including human cell lines. Its application to human induced pluripotent stem cells (hiPSCs) allows for the generation of isogenic cell pairs that differ in a single genetic lesion, and therefore the identification and characterization of causal genetic variants. We describe a simple, effective approach to perform delicate manipulations of the genome of hiPSCs through delivery of Cas9 RNPs along with ssDNA oligonucleotide repair templates that can generate mutations in up to 98% of single cell clones and introduce single nucleotide changes at an efficiency of up to 40%. We describe our use of a T7 endonuclease assay to identify active guide RNAs, and a high-throughput sequencing genotyping strategy that allows the identification of correctly edited clones. We also present our experiences of generating single nucleotide changes at 15 sites, which show considerable variability between both guides and target sites in the efficiency at which such changes can be introduced.

GPR35 promotes glycolysis, proliferation, and oncogenic signaling by engaging with the sodium potassium pump.

The sodium potassium pump (Na/K-ATPase) ensures the electrochemical gradient of a cell through an energy-dependent process that consumes about one-third of regenerated ATP. We report that the G protein-coupled receptor GPR35 interacted with the α chain of Na/K-ATPase and promotes its ion transport and Src signaling activity in a ligand-independent manner. Deletion of Gpr35 increased baseline Ca2+ to maximal levels and reduced Src activation and overall metabolic activity in macrophages and intestinal epithelial cells (IECs). In contrast, a common T108M polymorphism in GPR35 was hypermorphic and had the opposite effects to Gpr35 deletion on Src activation and metabolic activity. The T108M polymorphism is associated with ulcerative colitis and primary sclerosing cholangitis, inflammatory diseases with a high cancer risk. GPR35 promoted homeostatic IEC turnover, whereas Gpr35 deletion or inhibition by a selective pepducin prevented inflammation-associated and spontaneous intestinal tumorigenesis in mice. Thus, GPR35 acts as a central signaling and metabolic pacesetter, which reveals an unexpected role of Na/K-ATPase in macrophage and IEC biology.

Generation of gene-corrected human induced pluripotent stem cell lines derived from retinitis pigmentosa patient with Ser331Cysfs*5 mutation in MERTK.

The human induced pluripotent stem cell (hiPSC) line RP1-FiPS4F1 generated from the patient with autosomal recessive retinitis pigmentosa (arRP) caused by homozygous Ser331Cysfs*5 mutation in Mer tyrosine kinase receptor (MERTK) was genetically corrected using CRISPR/Cas9 system. Two isogenic hiPSCs lines, with heterozygous and homozygous correction of c.992_993delCA mutation in the MERTK gene were generated. These cell lines demonstrate normal karyotype, maintain a pluripotent state, and can differentiate toward three germ layers in vitro. These genetically corrected hiPSCs represent accurate controls to study the contribution of the specific genetic change to the disease, and potentially therapeutic material for cell-replacement therapy.

White Chick Syndrome Associated with Chicken Astrovirus in Ontario, Canada.

Sixty-four cases of white chick syndrome (WCS) in broiler breeders producing affected progeny were reported from seven hatcheries in Ontario, Canada, between 2009 and 2016, with 43 of those originating from two hatcheries owned by a single company. WCS cases were identified by the presence of typical chicks in the hatchery that were generally weak with pale to white down, enlarged abdomens, and occasionally brown wiry fluff on the dorsum of the neck. Affected embryos and chicks had characteristic gross and histologic liver lesions, and livers were positive for chicken astrovirus (CAstV) RNA by real-time reverse transcriptase PCR. Affected broiler breeder flocks experienced egg production drops of 0% to 21% and hatchability drops of 0% to 68.4%. The amino acid sequence of the region encoding the capsid gene of WCS viruses demonstrated all Ontario CAstV to be in Group B, Subgroup Bii.

Intermembrane crosstalk drives inner-membrane protein organization in Escherichia coli.

Gram-negative bacteria depend on energised protein complexes that connect the two membranes of the cell envelope. However, ß-barrel outer-membrane proteins (OMPs) and α-helical inner-membrane proteins (IMPs) display quite different organisation. OMPs cluster into islands that restrict their lateral mobility, while IMPs generally diffuse throughout the cell. Here, using live cell imaging of Escherichia coli, we demonstrate that when transient, energy-dependent transmembrane connections are formed, IMPs become subjugated by the inherent organisation of OMPs and that such connections impact IMP function. We show that while establishing a translocon for import, the colicin ColE9 sequesters the IMPs of the proton motive force (PMF)-linked Tol-Pal complex into islands mirroring those of colicin-bound OMPs. Through this imposed organisation, the bacteriocin subverts the outer-membrane stabilising role of Tol-Pal, blocking its recruitment to cell division sites and slowing membrane constriction. The ordering of IMPs by OMPs via an energised inter-membrane bridge represents an emerging functional paradigm in cell envelope biology.

Economic Impacts of White Chick Syndrome in Ontario, Canada.

A cluster of 12 cases of White Chick Syndrome (WCS) in broiler breeder flocks producing affected progeny occurred from June to November 2015 in two broiler chicken hatcheries owned by a single company in Ontario, Canada. Cases were identified by the presence of typical chicks in the hatchery characterized by pale to white down, enlarged abdomens, and occasionally brown wiry fluff on the dorsum of the neck that were generally weak. Affected broiler breeder flocks experienced egg production drops of 0% to 15% and hatchability drops of 1.8% to 49.1%. Some flocks experienced increased feed clean-up duration and/or reduced hatching egg weight. The financial impacts of WCS to affected hatching egg producers averaged $5,912 CAD (US$4,417) per 10 000 hens and were as great as $16,788 CAD (US$12,544) per 10 000 hens. The financial impacts of WCS to the affected hatcheries averaged $1,723 CAD (US$1,287) per 10 000 broiler breeder hens and were as great as $4,096 (US$3,060) per 10 000 hens.

Factors related to nurse comfort when caring for families experiencing perinatal loss: evidence for bereavement program enhancement.

As nurses provide holistic support, their own comfort in caring for parents and families experiencing perinatal loss must be considered. Study results showed that, although education is essential, experience independently predicted comfort in delivering perinatal bereavement care. Evidence from this study promotes the discussion of how nurse educators can structure professional development programs to best transfer the experience and confidence of perinatal nurses who are already comfortable with bereavement care to nurses who are not.

The impact of weight loss among seniors on Medicare spending.

OBJECTIVE: To examine the impact of temporary and permanent weight loss of 10% and 15% on 10-year and lifetime Medicare spending among adults with overweight and obesity aged 65 years and older. Weight loss of this magnitude is consistent with next generation anti-obesity medications recently approved by the Food and Drug Administration. METHODS: We follow the approach of a longitudinal dynamic aging process model developed by our research team. This model considers the dynamic relationships between weight, chronic disease, acute medical events, functional status, mortality, health care utilization and spending among Medicare beneficiaries from age 65 until death. Using this model, we estimate baseline Medicare spending over the next decade and then over the lifetime of seniors with a body mass index (BMI) ≥ 27 with at least one weight-related comorbidity (overweight), and seniors with obesity having a BMI ≥ 30 and ≥ 35. We then estimate Medicare spending for this population between ages 65 and 70 over the course of a year, assuming 10% and 15% weight loss under alternative scenarios: with and without weight regain. (Weight regain is assumed to be 90% over a 10-year period.) The difference in spending between baseline (no weight-loss intervention) and the alternative scenarios represent potential gross savings to the Medicare program. RESULTS: Permanent weight loss of 10 to 15% will yield $9,445 to $15,987 in gross per capita savings throughout their lifetime, and $8,070 to $13,474 over ten years. Similarly, initial weight loss of 10 to 15% followed by 90% weight regain will result in gross per capita savings of $7,556 to $11,109 over their lifetime, and $6,456 to $8,911 over ten years. Targeting weight loss medications to adults with obesity (BMI ≥ 30) produces greater savings to the Medicare program. CONCLUSION: Medicare can realize significant cost savings through anti-obesity medications that produce substantial weight loss, and as a result, reduce the progression to type 2 diabetes, and improve blood pressure and glycemic indicators in hypertensive and diabetic patients, respectively. Medications are currently excluded from coverage in the Medicare program, however, in light of potential savings and health benefits, may warrant consideration.

Nursing PhD consortia: a model for maximizing scarce resources.

Doctoral (PhD) education in nursing is costly and requires scarce resources: qualified faculty, qualified students, research funding, and infrastructure. This article discusses the development and implementation of a five-school consortium for delivery of an established PhD in Nursing Science program throughout north Florida. Factors that contributed to the success of the Consortium, including communication, history of shared work, collaborative approaches, and a formal agreement, are described. Challenges, such as maintaining curricular integrity across settings and selecting web-based formats, are considered. Results to date have been a viable consortium with a 4-year history, three PhD consortium graduates, 22 PhD students enrolled via the consortium, and success in attracting both federal and private funding. Consortia are proposed as a strategy for the effective use of limited resources, and suggestions are provided for the development of successful consortium models capable of delivering high-quality PhD nursing education.