Induction of Nitric Oxide Synthase in the J774.7 Macrophage Cell Line

When used alone, LPS and IFN-gamma were weak stimuli for inducing production of nitrite by J774.7 macrophages, but when used together, a powerful synergistic rise in nitrite accumulation was seen. Nitrite was derived from the L-arginine/NO pathway since its production was reduced by the NO synthase inhibitors, L-NMMA and L-NAME. L-NMMA was much more effective than L-NAME but the reason for this was obscure. The peptide polymyxin B also reduced nitrite accumulation by inhibiting the action of LPS on the cells. It is likely that NO was synthesised by the inducible form of NO synthase since nitrite production was low in the absence of stimuli and was reduced by dexamethasone in the presence of stimuli. Dexamethasone reduced nitrite accumulation, though whether via a direct action on the transcription process or through the formation of lipocortin 1 is unknown. Elevation of cyclic AMP levels reduced the production of nitrite by the cells, but only by a maximum of around 30%. Most agents had little effect on nitrite accumulation when added after LPS and IFN-gamma, and produced most of their effects when added before the induction of NO synthase by LPS and IFN-gamma. Thus, cyclic AMP can only regulate the induction process, probably at the level of transcription, to a slight degree. Elevation of cyclic GMP levels reduced accumulation of nitrite but only by around 30%. The effects of GTN may be mediated both via toxic effects on the NO synthase enzyme by NO and via a cyclic GMP-dependent mechanism. It is unknown if elevated cyclic GMP levels can induce phosphorylation of the NO synthase enzyme and so modulate its activity. It appears that both PKC and tyrosine kinase play a role in the induction of NO synthase by LPS and IFN-gamma, although evidence for the latter is more straightforward. The smooth muscle relaxant released from J774.7 cells following stimulation by LPS and EIFN-gamma was sensitive to attack from superoxide anions and blocked by haemoglobin. Furthermore, production of the relaxant was blocked following incubation with the inhibitor of NO synthase, L-NMMA. Thus, in all respects, this relaxant substance behaves like free NO. The mechanism by which macrophages protect themselves from the toxic effects of the high concentrations of NO they produce is unknown. The release of superoxide anions from J774.7 cells was undetectable, although the cells appear to release spontaneously a powerful oxidant. The nature of this oxidant is unknown, but it appears not to be hydrogen peroxide, peroxynitrite, hydroxyl radical or hypochlorous acid

Advancing Cancer Prevention Practice Facilitation Work in Rural Primary Care During COVID-19

COVID-19 and the response to slow the virus spread in West Virginia (WV), including a statewide stay-at-home order, presented challenges to rural primary care clinics on the frontlines. These challenges affected critical quality improvement work, including cancer screening services. In this commentary, the authors present the results of a survey of WV primary care practices that highlight potential long-term implications and identifies opportunities for practice facilitators to partner with rural primary care clinics to address them

Equine Multiple Congenital Ocular Anomalies maps to a 4.9 megabase interval on horse chromosome 6

<p>Abstract</p> <p>Background</p> <p>Equine Multiple Congenital Ocular Anomalies (MCOA) syndrome consists of a diverse set of abnormalities predominantly localized to the frontal part of the eye. The disease is in agreement with a codominant mode of inheritance in our horse material. Animals presumed to be heterozygous for the mutant allele have cysts originating from the temporal ciliary body, peripheral retina and/or iris. In contrast, animals predicted to be homozygous for the disease-causing allele possess a wide range of multiple abnormalities, including iridociliary and/or peripheral retinal cysts, iridocorneal angle abnormalities, cornea globosa, iris hypoplasia and congenital cataracts. MCOA is most common in the Rocky Mountain horse breed where it occurs at a high frequency among Silver colored horses. The Silver coat color is associated with mutations in <it>PMEL17 </it>that resides on ECA6q23. To map the <it>MCOA </it>locus we analyzed 11 genetic markers on ECA6q and herein describe a chromosome interval for the <it>MCOA </it>locus.</p> <p>Results</p> <p>We performed linkage analysis within 17 paternal half-sib families of the Rocky Mountain horse breed. More than half of the 131 offspring had the Cyst phenotype and about one third had MCOA. Segregation data were obtained by genotyping 10 microsatellite markers most of which are positioned on ECA6q22-23, as well as the missense mutation for the Silver phenotype in <it>PMEL17</it>. Significant linkage was found between the <it>MCOA </it>locus and eight of the genetic markers, where marker <it>UPP5 </it>(Theta = 0, z = 12.3), <it>PMEL17ex11 </it>(Theta = 0, z = 19.0) and <it>UPP6 </it>(Theta = 0, z = 17.5) showed complete linkage with the <it>MCOA </it>locus. DNA sequencing of <it>PMEL17 </it>in affected and healthy control individuals did not reveal any additional mutations than the two mutations associated with the Silver coat color.</p> <p>Conclusion</p> <p>The <it>MCOA </it>locus can with high confidence be positioned within a 4.9 megabase (Mb) interval on ECA6q. The genotype data on <it>UPP5</it>, <it>PMEL17ex11 </it>and <it>UPP6 </it>strongly support the hypothesis that horses with the Cyst phenotype are heterozygous for the mutant allele and that horses with the MCOA phenotype are homozygous for the mutant allele.</p

Case Study of a Comprehensive Team-Based Approach to Increase Colorectal Cancer Screening

Introduction: Colorectal cancer is the second leading cause of cancer deaths among men and women in West Virginia. In addition, 51% of all colorectal cancers diagnosed in West Virginia from 2012 to 2016 were detected at either regional (31%) or distant (20%) stages indicating a need for improved early detection. Methods: West Virginia University Cheat Lake Physicians participated in the West Virginia Program to Increase Colorectal Cancer Screening, a program of Cancer Prevention and Control at the WVU Cancer Institute. As a result, Cheat Lake Physicians assembled a team of health care professionals to implement evidence-based interventions and system changes including provider assessment and feedback, patient reminders, accurate data capture, and tracking of CRC screening tests. Results: These efforts resulted in a 15.8% increase in colorectal cancer screening rates within one year of implementation. Additionally, the clinic achieved a 66% return rate for Fecal Immunochemical Test kits, an inexpensive, stool-based colorectal cancer screening test. Implications: The utilization of a team-based approach to patient care yields positive results that can be carried over to other cancer and disease prevention efforts in primary care clinics

Leveraging Electronic Health Records Data for Enhanced Colorectal Cancer Screening Efforts

Introduction: Colorectal cancer is the third most common type of cancer in the United States for men and women combined. While the current threat of disease nationally is significant, the majority of colorectal cancer cases and deaths could be prevented through established screening tests and guidelines. Within the Appalachian region and West Virginia in particular, colorectal cancer is a significant public health problem. A more systematic, comprehensive approach to preventing and controlling cancer is essential. Methods: Through the West Virginia Program to Increase Colorectal Cancer Screening, primary care systems across the state received data-informed practice facilitation designed to increase screening rates. Results: Year-1 cohort health systems had an overall baseline screening rate of 28.4% during calendar year 2014. This rate increased and remained steady during the three follow-up measurement time periods, with a rate of 49.5% during calendar year 2018. This increase is notably greater than comparable health systems not part of the initiative. Implications: Lessons learned in increasing colorectal cancer screening rates are applicable to other priority health needs as well

Driving digital health transformation in hospitals:a formative qualitative evaluation of the English Global Digital Exemplar programme

BACKGROUND: There is currently a strong drive internationally towards creating digitally advanced healthcare systems through coordinated efforts at a national level. The English Global Digital Exemplar (GDE) programme is a large-scale national health information technology change programme aiming to promote digitally-enabled transformation in secondary healthcare provider organisations by supporting relatively digitally mature provider organisations to become international centres of excellence. AIM: To qualitatively evaluate the impact of the GDE programme in promoting digital transformation in provider organisations that took part in the programme. METHODS: We conducted a series of in-depth case studies in 12 purposively selected provider organisations and a further 24 wider case studies of the remaining organisations participating in the GDE programme. Data collected included 628 interviews, non-participant observations of 190 meetings and workshops and analysis of 9 documents. We used thematic analysis aided by NVivo software and drew on sociotechnical theory to analyse the data. RESULTS: We found the GDE programme accelerated digital transformation within participating provider organisations. This acceleration was triggered by: (1) dedicated funding and the associated requirement for matched internal funding, which in turn helped to prioritise digital transformation locally; (2) governance requirements put in place by the programme that helped strengthen existing local governance and project management structures and supported the emergence of a cadre of clinical health informatics leaders locally; and (3) reputational benefits associated with being recognised as a centre of digital excellence, which facilitated organisational buy-in for digital transformation and increased negotiating power with vendors. CONCLUSION: The GDE programme has been successful in accelerating digital transformation in participating provider organisations. Large-scale digital transformation programmes in healthcare can stimulate local progress through protected funding, putting in place governance structures and leveraging reputational benefits for participating provider organisations, around a coherent vision of transformation

Interorganizational Knowledge Sharing to Establish Digital Health Learning Ecosystems:Qualitative Evaluation of a National Digital Health Transformation Program in England

BACKGROUND: The English Global Digital Exemplar (GDE) program is one of the first concerted efforts to create a digital health learning ecosystem across a national health service. OBJECTIVE: This study aims to explore mechanisms that support or inhibit the exchange of interorganizational digital transformation knowledge. METHODS: We conducted a formative qualitative evaluation of the GDE program. We used semistructured interviews with clinical, technical, and managerial staff; national program managers and network leaders; nonparticipant observations of knowledge transfer activities through attending meetings, workshops, and conferences; and documentary analysis of policy documents. The data were thematically analyzed by drawing on a theory-informed sociotechnical coding framework. We used a mixture of deductive and inductive methods, supported by NVivo software, to facilitate coding. RESULTS: We conducted 341 one-on-one and 116 group interviews, observed 86 meetings, and analyzed 245 documents from 36 participating provider organizations. We also conducted 51 high-level interviews with policy makers and vendors; performed 77 observations of national meetings, workshops, and conferences; and analyzed 80 national documents. Formal processes put in place by the GDE program to initiate and reinforce knowledge transfer and learning have accelerated the growth of informal knowledge networking and helped establish the foundations of a learning ecosystem. However, formal networks were most effective when supported by informal networking. The benefits of networking were enhanced (and costs reduced) by geographical proximity, shared culture and context, common technological functionality, regional and strategic alignments, and professional agendas. CONCLUSIONS: Knowledge exchange is most effective when sustained through informal networking driven by the mutual benefits of sharing knowledge and convergence between group members in their organizational and technological setting and goals. Policy interventions need to enhance incentives and reduce barriers to sharing across the ecosystem, be flexible in tailoring formal interventions to emerging needs, and promote informal knowledge sharing