The ilv2 gene, encoding acetolactate synthase for branched chain amino acid biosynthesis, is required for plant pathogenicity by Leptosphaeria maculans.

BACKGROUND: Control of blackleg disease of canola caused by the fungus Leptosphaeria maculans relies on strategies such as the inhibition of growth with fungicides. However, other chemicals are used during canola cultivation, including fertilizers and herbicides. There is widespread use of herbicides that target the acetolactate synthase (ALS) enzyme involved in branched chain amino acid synthesis and low levels of these amino acids within leaves of Brassica species. In L. maculans the ilv2 gene encodes ALS and thus ALS-inhibiting herbicides may inadvertently impact the fungus. METHODS AND RESULTS: Here, the impact of a commercial herbicide targeting ALS and mutation of the homologous ilv2 gene in L. maculans was explored. Exposure to herbicide had limited impact on growth in vitro but reduced lesion sizes in plant disease experiments. Furthermore, the mutation of the ilv2 gene via CRISPR-Cas9 gene editing rendered the fungus non-pathogenic. CONCLUSION: Herbicide applications can influence disease outcome, but likely to a minor extent.

Genetic Transformation of Cryptococcus Species with Agrobacterium Transfer DNA.

Transformation of foreign DNA into Cryptococcus species is a powerful tool for exploring gene functions in these human pathogens. Agrobacterium tumefaciens-mediated transformation (AtMT) has been used for the stable introduction of exogenous DNA into Cryptococcus for over two decades, being particularly impactful for insertional mutagenesis screens to discover new genes involved in fungal biology. A detailed protocol to conduct this transformation method is provided in the chapter. Scope for modifications and the benefits and disadvantages of using AtMT in Cryptococcus species are also presented.

Comparison of ChatGPT knowledge against 2020 consensus statement on ankyloglossia in children.

OBJECTIVE: This paper evaluates ChatGPT's accuracy and consistency in providing information on ankyloglossia, a congenital oral condition. Assessing alignment with expert consensus, the study explores potential implications for patients relying on AI for medical information. METHODS: Statements from the 2020 clinical consensus statement on ankyloglossia were presented to ChatGPT, and its responses were scored using a 9-point Likert scale. The study analyzed the mean and standard deviation of ChatGPT scores for each statement. Statistical analysis was conducted using Excel. RESULTS: Among the 63 statements assessed, 67 % of ChatGPT responses closely aligned with expert consensus mean scores. However, 17 % (11/63) were statements in which the ChatGPT mean response was different from the CCS mean by 2.0 or greater, raising concerns about ChatGPT's potential influence in disseminating uncertain or debated medical information. Variations in mean scores highlighted discrepancies, with some statements showing significant deviations from expert opinions. CONCLUSION: While ChatGPT mirrored medical viewpoints on ankyloglossia, alignment with non-consensus statements raises caution in relying on it for medical advice. Future research should refine AI models, address inaccuracies, and explore diverse user queries for safe integration into medical decision-making. Despite potential benefits, ongoing examination of ChatGPT's power and limitations is crucial, considering its impact on health equity and information access.

"Ultraviolet C phototherapy as an adjunct wound therapy in full-thickness wound of a rat model".

Ultraviolet C (UVC) phototherapy is a potential modality to improve wound healing due to its well-known antimicrobial properties, and it promotes the expression of certain growth factors. However, limited data exist to show the clinical effect of UVC in wound healing compared with other advanced modern dressings. This animal preclinical study involved 56 Sprague-Dawley female rats aged 3 months old, weighing between 250 and 300 g, which were acclimatized for one week. Following the creation of a 2 × 2-cm-square full-thickness wound over the dorsum of each rat, they were divided into four treatment groups, namely, the control, UVC, hydrofiber silver, and UVC/hydrofiber silver groups. On Days 2, 4, 7, 10, 14, 21, and 28 postwounding, two rats from each group underwent wound assessment via wound measurement (mm2), calculation of the percentage of wound contracture and percentage of epithelization. Wound specimens were obtained for histological examination of inflammatory cells (neutrophils, lymphocytes and macrophages) and fibroblast cell counts. There was a relationship between wound size reduction and time to heal (P<0.05, R2=0.70) among the four treatment groups. The UVC/hydrofiber silver group had a significantly smaller wound size given the time to heal compared with the control group (P = 0.01) and UVC group (P = 0.02). There were no significant differences in terms of wound contracture and epithelization percentage among the four treatment groups. Histopathological examination revealed a significantly lower mean fibroblast count in the UVC/hydrofiber silver group than in the other groups (P = 0.025). These data suggested that UVC phototherapy did not increase the rate of healing but maintained the integrity of the wound by providing antimicrobial properties and preventing overproduction of fibroblasts. UVC was also safe, as no overt inflammatory response was discovered.

A large transposable element mediates metal resistance in the fungus Paecilomyces variotii.

The horizontal transfer of large gene clusters by mobile elements is a key driver of prokaryotic adaptation in response to environmental stresses. Eukaryotic microbes face similar stresses; however, a parallel role for mobile elements has not been established. A stress faced by many microorganisms is toxic metal ions in their environment. In fungi, identified mechanisms for protection against metals generally rely on genes that are dispersed within an organism's genome. Here, we discover a large (∼85 kb) region that confers tolerance to five metal/metalloid ions (arsenate, cadmium, copper, lead, and zinc) in the genomes of some, but not all, strains of a fungus, Paecilomyces variotii. We name this region HEPHAESTUS (Hφ) and present evidence that it is mobile within the P. variotii genome with features characteristic of a transposable element. HEPHAESTUS contains the greatest complement of host-beneficial genes carried by a transposable element in eukaryotes, suggesting that eukaryotic transposable elements might play a role analogous to bacteria in the horizontal transfer of large regions of host-beneficial DNA. Genes within HEPHAESTUS responsible for individual metal tolerances include those encoding a P-type ATPase transporter-PcaA-required for cadmium and lead tolerance, a transporter-ZrcA-providing tolerance to zinc, and a multicopper oxidase-McoA-conferring tolerance to copper. In addition, a subregion of Hφ confers tolerance to arsenate. The genome sequences of other fungi in the Eurotiales contain further examples of HEPHAESTUS, suggesting that it is responsible for independently assembling tolerance to a diverse array of ions, including chromium, mercury, and sodium.

Community Health Center Residency Training: Improving Staffing, Service, and Quality.

BACKGROUND AND OBJECTIVES: Community-based residency programs are an important strategy to address rural and underserved primary care shortages, however, health centers report both benefits and challenges to training. This study aims to understand the impact of new Teaching Health Center (THC) residency programs on health center staffing, patient service, quality of care, and provider productivity. METHODS: Using the Uniform Data System, we used inverse propensity score weighting to create a balanced sample of new THC and non-THC health centers in 2010. Using 2018 data, we applied propensity score weighted regressions to examine changes in staffing, service, quality of care, and productivity in THC versus non-THC health centers. RESULTS: In 2018, health centers with new THC programs were associated with increased physician (16.40, P<.01) staffing, yet decreased physician visits per full-time equivalent (-425.3, P<.01) relative to non-THC centers. New THC centers had increased delivery visits (231.0, P<.05), and had a greater rate of early entry into prenatal care (4.90%, P<.01). CONCLUSIONS: New residency programs are associated with increased provider recruitment, expanded patient service, and some improved health outcomes, but also with potential decreased provider productivity in health centers.

Coronavirus Disease 2019 Planning and Response: A Tale of 2 Health Workforce Estimator Tools.

BACKGROUND: As coronavirus disease 2019 (COVID-19) rapidly progressed throughout the United States, increased demand for health workers required health workforce data and tools to aid planning and response at local, state, and national levels. OBJECTIVE: We describe the development of 2 estimator tools designed to inform health workforce planning for COVID-19. RESEARCH DESIGN: We estimated supply and demand for intensivists, critical care nurses, hospitalists, respiratory therapists, and pharmacists, using Institute for Health Metrics and Evaluation projections for COVID-19 hospital care and National Plan and Provider Enumeration System, Provider Enrollment Chain and Ownership System, American Hospital Association, and Bureau of Labor Statistics Occupation Employment Statistics for workforce supply. We estimated contact tracing workforce needs using Johns Hopkins University COVID-19 case counts and workload parameters based on expert advice. RESULTS: The State Hospital Workforce Deficit Estimator estimated the sufficiency of state hospital-based clinicians to meet projected COVID-19 demand. The Contact Tracing Workforce Estimator calculated the workforce needed based on the 14-day COVID-19 caseload at county, state, and the national level, allowing users to adjust workload parameters to reflect local contexts. CONCLUSIONS: The 2 estimators illustrate the value of integrating health workforce data and analysis with pandemic response planning. The many unknowns associated with COVID-19 required tools to be flexible, allowing users to change assumptions on number of contacts and work capacity. Data limitations were a challenge for both estimators, highlighting the need to invest in health workforce data and data infrastructure as part of future emergency preparedness planning.

The Use of Community Health Workers in Community Health Centers.

BACKGROUND: Until 2016, community health centers (CHCs) reported community health workers (CHWs) as part of their overall enabling services workforce, making analyses of CHW use over time infeasible in the annual Uniform Data System (UDS). OBJECTIVE: The objective of this study was to examine changes in the CHW workforce among CHCs from 2016 to 2018 and factors associated with the use of CHWs. RESEARCH DESIGN, SUBJECTS, MEASURES: The two-part model estimated separate effects for the probability of using any CHW and extent of CHW full-time equivalents (FTEs) reported in those CHCs, using a total of 4102 CHC-year observations from 2016 to 2018. To estimate the extent to which increases in CHW workforce are attributable to real growth or rather are a consequence of a change in reporting category, we also conducted a difference-in-differences analysis to compare non-CHW enabling services FTEs between CHCs with and without CHWs before (2013-2015) and after (2016-2018) the reporting change in 2016. RESULTS: The rate of CHCs that employed CHWs rose from 20.04% in 2016 to 28.34% in 2018, while average FTEs stayed relatively flat (3.32 FTEs). Patient visit volume (larger CHCs) and grant funding (less reliant on federal but more reliant on private funding) were significant factors associated with CHW use. However, we found that a substantial portion of this growth was attributable to a change in UDS reporting categories. CONCLUSION: While we do not address the reasons why CHCs have been slow to use CHWs, our results point to substantial financial barriers associated with CHCs' expanding the use of CHWs.

Independent freestanding emergency departments and implications for the rural emergency physician workforce in Texas.

OBJECTIVE: Independent freestanding emergency departments (IFEDs) have proliferated over the last decade, largely in Texas. We examined the IFED physician workforce composition and changes in emergency physician workforce supply across states and in rural Texas over the period of IFED proliferation following a 2009 legislation allowing the licensing of these sites. DATA SOURCES: IFED websites, Texas Medical Board lookup tool, National Plan & Provider Enumeration System (NPPES), Provider Enrollment and Chain/Ownership System (PECOS), Medicare Physician Shared Patient Patterns, CareSet DocGraph Hop Teaming, Healthcare Provider Database. STUDY DESIGN: Descriptive analysis of the IFED physician workforce; quasi-experimental difference-in-difference analysis of Texas emergency physician movement into and out of the state; and difference-in-difference-in-difference analysis of the change in emergency physician supply between rural and urban areas in Texas compared with other states. DATA EXTRACTION METHODS: Using the NPIs obtained through Texas IFED websites and Texas Medical Board data, we examined NPPES/PECOS files, Medicare Physician Shared Patient Patterns, and CareSet DocGraph Hop Teaming for IFED physician practice locations from 2009 to 2017. We extracted all active emergency physicians from a Healthcare Provider Database, derived from a 5% Medicare claims (1999-2017). PRINCIPAL FINDINGS: In 2019, 545 physicians practiced in Texas IFEDs, of which 515 (94.5%) were emergency physicians. We located 533 in previous practice, of whom 522 (97.9%) previously practiced in Disproportionate Share Hospitals and 100 (18.8%) in rural areas. Following legislation to begin licensing IFEDs in 2009, there were on average 42.1 (P < .01) moving into Texas and 17.0 (P < .01) fewer moving out compared with all other states. Our results also indicated that the difference in emergency physician supply between rural and urban Texas was 1,002 (P < .01) fewer than for other states. CONCLUSIONS: New models of health care organizations such as IFEDs have workforce implications that may further exacerbate rural and underserved workforce and access challenges.

Constant-potential environment for activating and synchronizing cardiomyocyte colonies with on-chip ion-depleting perm-selective membranes.

In this study, an ion depleted zone created by an ion-selective membrane was used to impose a high and uniform constant extracellular potential over an entire ∼1000 cell rat cardiomyocyte (rCM) colony on-a-chip to trigger synchronized voltage-gated ion channel activities while preserving cell viability, thus extending single-cell voltage-clamp ion channel studies to an entire normalized colony. Image analysis indicated that rCM beating was strengthened and accelerated (by a factor of ∼2) within minutes of ion depletion and the duration of contraction and relaxation phases was significantly reduced. After the initial synchronization, the entire colony responds collectively to external potential changes such that beating over the entire colony can be activated or deactivated within 0.1 s. These newly observed collective dynamic responses, due to simultaneous ion channel activation/deactivation by a uniform constant-potential extracellular environment, suggest that perm-selective membrane modules on cell culture chips can facilitate studies of extracellular cardiac cell electrical communication and how ion-channel related pathologies affect cardiac cell synchronization. The future applications of this new technology can lead to better drug screening platforms for cardiotoxicity as well as platforms that can facilitate synchronized maturation of pluripotent stem cells into colonies with high electrical connectivity.

WWOSC 2014: research needs for better health resilience to weather hazards.

The first World Weather Open Science Conference (WWOSC, held from 17-21 August 2014 in Montreal, Québec), provided an open forum where the experience and perspective of a variety of weather information providers and users was combined with the latest application advances in social sciences. A special session devoted to health focused on how best the most recent weather information and communication technologies (ICT) could improve the health emergency responses to disasters resulting from natural hazards. Speakers from a plenary presentation and its corresponding panel shared lessons learnt from different international multidisciplinary initiatives against weather-related epidemics, such as malaria, leptospirosis and meningitis and from public health responses to floods and heat waves such as in Ontario and Quebec, Canada. Participants could bear witness to recent progress made in the use of forecasting tools and in the application of increased spatiotemporal resolutions in the management of weather related health risks through anticipative interventions, early alert and warning and early responses especially by vulnerable groups. There was an agreement that resilience to weather hazards is best developed based on evidence of their health impact and when, at local level, there is a close interaction between health care providers, epidemiologists, climate services, public health authorities and communities. Using near real time health data (such as hospital admission, disease incidence monitoring…) combined with weather information has been recommended to appraise the relevance of decisions and the effectiveness of interventions and to make adjustments when needed. It also helps appraising how people may be more or less vulnerable to a particular hazard depending on the resilience infrastructures and services. This session was mainly attended by climate, environment and social scientists from North American and European countries. Producing a commentary appears to be an effective way to share this session's conclusions to research institutions and public health experts worldwide. It also advocates for better linking operational research and decision making and for appraising the impact of ICT and public health interventions on health.

Isolation of human lymphatic malformation endothelial cells, their in vitro characterization and in vivo survival in a mouse xenograft model.

Human lymphatic vascular malformations (LMs), also known as cystic hygromas or lymphangioma, consist of multiple lymphatic endothelial cell-lined lymph-containing cysts. No animal model of this disease exists. To develop a mouse xenograft model of human LM, CD34(Neg)CD31(Pos) LM lymphatic endothelial cells (LM-LEC) were isolated from surgical specimens and compared to foreskin CD34(Neg)CD31(Pos) lymphatic endothelial cells (LECs). Cells were implanted into a mouse tissue engineering model for 1, 2 and 4 weeks. In vitro LM-LECs showed increased proliferation and survival under starvation conditions (P < 0.0005 at 48 h, two-way ANOVA), increased migration (P < 0.001, two-way ANOVA) and formed fewer (P = 0.029, independent samples t test), shorter tubes (P = 0.029, independent samples t test) than foreskin LECs. In vivo LM-LECs implanted into a Matrigel™-containing mouse chamber model assembled to develop vessels with dilated cystic lumens lined with flat endothelium, morphology similar to that of clinical LMs. Human foreskin LECs failed to survive implantation. In LM-LEC implanted chambers the percent volume of podoplanin(Pos) vessels was 1.18 ± 2.24 % at 1 week, 6.34 ± 2.68 % at 2 weeks and increasing to 7.67 ± 3.60 % at 4 weeks. In conclusion, the significantly increased proliferation, migration, resistance to apoptosis and decreased tubulogenesis of LM-LECs observed in vitro is likely to account for their survival and assembly into stable LM-like structures when implanted into a mouse vascularised chamber model. This in vivo xenograft model will provide the basis of future studies of LM biology and testing of potential pharmacological interventions for patients with lymphatic malformations.

Non-physician clinician provided HIV treatment results in equivalent outcomes as physician-provided care: a meta-analysis.

INTRODUCTION: A severe healthcare worker shortage in sub-Saharan Africa is inhibiting the expansion of HIV treatment. Task shifting, the transfer of antiretroviral therapy (ART) management and initiation from doctors to nurses and other non-physician clinicians, has been proposed to address this problem. However, many health officials remain wary about implementing task shifting policies due to concerns that non-physicians will provide care inferior to physicians. To determine if non-physician-provided HIV care does result in equivalent outcomes to physician-provided care, a meta-analysis was performed. METHODS: Online databases were searched using a predefined strategy. The results for four primary outcomes were combined using a random effects model with sub-groups of non-physician-managed ART and -initiated ART. TB diagnosis rates, adherence, weight gain and patient satisfaction were summarized qualitatively. RESULTS: Mortality (N=59,666) had similar outcomes for non-physicians and physicians, with a hazard ratio of 1.05 (CI: 0.88-1.26). The increase in CD4 levels at one year, as a difference in means of 2.3 (N=17,142, CI: -12.7-17.3), and viral failure at one year, as a risk ratio of 0.89 (N=10,344, CI: 0.65-1.23), were similar for physicians and non-physicians. Interestingly, loss to follow-up (LTFU) (N=53,435) was reduced for non-physicians with a hazard ratio of 0.72 (CI: 0.56-0.94). TB diagnosis rates, adherence and weight gain were similar for non-physicians and physicians. Patient satisfaction appeared higher for non-physicians in qualitative components of studies and was attributed to non-physicians spending more time with patients as well as providing more holistic care. CONCLUSIONS: Non-physician-provided HIV care results in equivalent outcomes to care provided by physicians and may result in decreased LTFU rates.