Intracellular pH differentially regulates transcription of metabolic and signaling pathways in normal epithelial cells.

Intracellular pH (pHi) dynamics regulate normal cell function, and dysregulated pHi dynamics is an emerging hallmark of cancer (constitutively increased pHi) and neurodegeneration (constitutively decreased pHi). However, the molecular mechanisms by which pHi dynamics regulate cell biology are poorly understood. Here, we discovered that altering pHi in normal human breast epithelial cells triggers global transcriptional changes. We identified 176 genes differentially regulated by pHi, with pHi-dependent genes clustering in signaling and glycolytic pathways. Using various normal epithelial cell models, we showed pH-dependent Notch homolog 1 protein expression, with increased protein abundance at high pHi. This resulted in pH-dependent downstream signaling, with increased Notch homolog 1 signaling at high pHi. We also found that high pHi increased the expression of glycolytic enzymes and regulators of pyruvate fate, including lactate dehydrogenase and pyruvate dehydrogenase kinase. These transcriptional changes were sufficient to alter lactate production, with high pHi shifting these normal epithelial cells toward a glycolytic metabolism and increasing lactate production. Thus, pHi dynamics transcriptionally regulate signaling and metabolic pathways in normal epithelial cells. Our data reveal new molecular regulators of pHi-dependent biology and a role for increased pHi in driving the acquisition of cancer-associated signaling and metabolic changes in normal human epithelial cells.

Comparison of Alabama Nurse Experiences Between Practice Areas During the Early COVID-19 Pandemic.

BACKGROUND: According to the Total Worker Health® framework, safety culture including a reasonable workload among healthcare workers is essential to the security and well-being of patients, staff, and healthcare organizations. Evaluating the impact of the pandemic on the nursing workforce in different practice areas is critical for addressing workforce health and sustainability. The purpose of this study was to compare work and selfcare experiences among Alabama nurses between practice areas and the early pandemic years (2020 vs. 2021). METHODS: A secondary analysis of cross-sectional Alabama State Nurses Association (ASNA) survey data was conducted. Kruskal-Wallis analysis of variance, Wilcoxon rank, and false discovery rates were examined. RESULTS: There were 1,369 and 2,458 nurse survey responses in 2020 and 2021, respectively. By 2021, nurses reported worsening staff shortages, a greater need for retired and new graduate nurses to help with the workload burden, and perceptions of heavier emergency department workloads. Lower proportions of nurses reported the ability to engage in self-care activities and satisfaction with state and federal crisis management. Intensive care nurses were more likely to report staffing shortages while also reporting the lowest ability to engage in self-care. CONCLUSIONS: Overall, the Alabama nursing workforce perceived worsening work conditions in 2021 compared to when the pandemic began. Practice areas varied greatly in their responses, with acute and intensive care areas perceiving more difficult work conditions. Total Worker Health® programs should be designed to promote and support nurses' well-being based on their experience and the needs of specific practice areas.

KRT17High/CXCL8+ tumor cells display both classical and basal features and regulate myeloid infiltration in the pancreatic cancer microenvironment.

PURPOSE: Pancreatic ductal adenocarcinoma (PDAC) is generally divided in two subtypes, classical and basal. Recently, single cell RNA sequencing has uncovered the co-existence of basal and classical cancer cells, as well as intermediary cancer cells, in individual tumors. The latter remains poorly understood; here, we sought to characterize them using a multimodal approach. EXPERIMENTAL DESIGN: We performed subtyping on a single cell RNA sequencing dataset containing 18 human PDAC samples to identify multiple intermediary subtypes. We generated patient-derived PDAC organoids for functional studies. We compared single cell profiling of matched blood and tumor samples to measure changes in the local and systemic immune microenvironment. We then leveraged longitudinally patient-matched blood to follow individual patients over the course of chemotherapy. RESULTS: We identified a cluster of KRT17-high intermediary cancer cells that uniquely express high levels of CXCL8 and other cytokines. The proportion of KRT17High/CXCL8+ cells in patient tumors correlated with intra-tumoral myeloid abundance, and, interestingly, high pro-tumor peripheral blood granulocytes, implicating local and systemic roles. Patient-derived organoids maintained KRT17High/CXCL8+cells and induced myeloid cell migration in an CXCL8-dependent manner. In our longitudinal studies, plasma CXCL8 decreased following chemotherapy in responsive patients, while CXCL8 persistence portended worse prognosis. CONCLUSIONS: Through single cell analysis of PDAC samples we identified KRT17High/CXCL8+ cancer cells as an intermediary subtype, marked by a unique cytokine profile and capable of influencing myeloid cells in the tumor microenvironment and systemically. The abundance of this cell population should be considered for patient stratification in precision immunotherapy.

DNP Team Projects: A Reflection of Team Science.

BACKGROUND: Concern has been raised related to the rigor of DNP team projects due to the potential lack of individual opportunity for growth. However, team science, the scientific collaboration conducted by more than one individual in an interdependent fashion, is becoming standard practice for scientific inquiry and dissemination. DNP team projects provide an opportunity to demonstrate competencies related to collaboration, communication, organization, planning, reliability, accountability and acknowledgement of other opinions, expertise, and contributions. Faculty working with student teams may encounter challenges related to team dynamics and individual student evaluation. Thoughtful application of team science principles can assist in minimizing these challenges. METHOD: The purpose of this paper is to describe two school's combined experiences and lessons learned in application of team science to DNP team projects. CONCLUSION: When undertaken with an informed and organized approach, DNP team projects are an ideal strategy to enhance collaborative skills and position nurse leaders to positively impact health outcomes.

Multimodal Mapping of the Tumor and Peripheral Blood Immune Landscape in Human Pancreatic Cancer.

Pancreatic ductal adenocarcinoma (PDA) is characterized by an immune-suppressive tumor microenvironment that renders it largely refractory to immunotherapy. We implemented a multimodal analysis approach to elucidate the immune landscape in PDA. Using a combination of CyTOF, single-cell RNA sequencing, and multiplex immunohistochemistry on patient tumors, matched blood, and non-malignant samples, we uncovered a complex network of immune-suppressive cellular interactions. These experiments revealed heterogeneous expression of immune checkpoint receptors in individual patient's T cells and increased markers of CD8+ T cell dysfunction in advanced disease stage. Tumor-infiltrating CD8+ T cells had an increased proportion of cells expressing an exhausted expression profile that included upregulation of the immune checkpoint TIGIT, a finding that we validated at the protein level. Our findings point to a profound alteration of the immune landscape of tumors, and to patient-specific immune changes that should be taken into account as combination immunotherapy becomes available for pancreatic cancer.

Compliance with ivory trade regulations in the United Kingdom among traders.

Global demand for elephant ivory is contributing to illegal poaching and significant decline of African elephant (Loxondonta africana) populations. To help mitigate decline, countries with legal domestic ivory markets were recommended by the Convention on International Trade in Endangered Species of Wild Fauna and Flora to close domestic markets for commercial trade. However, implementing stricter regulations on wildlife trade does not necessarily mean compliance with rules will follow. Using an online questionnaire, we examined the relationship between self-reported compliance with ivory trade regulations among 115 ivory traders in the United Kingdom and 4 dimensions (control, deterrence, legitimacy, and social norms) hypothesized to influence compliance with conservation regulations. Although most traders supported regulations, a large number did not always check that they comply with them when trading objects containing ivory. The main factors influencing compliance with ivory trade regulations were traders' ability to comply and their perceptions of the regulations and punishments to deter illegal trade. These findings demonstrate the utility of conservation criminology to improve wildlife trade regulations and identify opportunities to reduce illegal ivory entering the market in the United Kingdom. Compliance could be improved by clearer regulations that facilitate easier detection of illegal ivory products and stronger prosecution of violations.

Cumplimiento de las Regulaciones del Mercado de Marfil entre Comerciantes del Reino Unido Resumen La demanda mundial por el marfil de elefante está contribuyendo a la caza ilegal y a la declinación significativa de la población del elefante africano (Loxondonta africana). Para ayudar a mitigar la declinación, la Convención sobre el Comercio Internacional de Especies Amenazadas de Fauna y Flora Silvestres recomendó a los países que tienen mercados domésticos legales de marfil que cierren estos mercados al intercambio comercial. Sin embargo, la implementación de regulaciones más estrictas sobre el mercado de fauna no significa necesariamente que después existirá un cumplimiento de las reglas establecidas. Examinamos con un cuestionario en línea la relación entre el cumplimiento auto-reportado y las regulaciones del mercado de marfil de 115 comerciantes de marfil en el Reino Unido y cuatro dimensiones (control, disuasión, legitimidad y normas sociales) que creemos influyen sobre el cumplimiento de las regulaciones de conservación. Aunque la mayoría de los comerciantes respaldó las regulaciones, un gran número no marcó en la encuesta si cumplían con ellas cuando comercializaban con marfil. Los factores principales que influyen sobre el cumplimiento de las regulaciones del mercado de marfil son la aptitud del comerciante para cumplir y las percepciones que tiene de las regulaciones y los castigos para disuadir el comercio ilegal. Estos resultados demuestran la utilidad de la criminología de la conservación para mejorar las regulaciones del comercio de fauna e identificar las oportunidades para reducir la entrada de marfil ilegal al mercado del Reino Unido. El cumplimiento de las regulaciones podría incrementarse por medio de regulaciones más claras que faciliten la detección de productos ilegales de marfil y seguimiento más fuerte de las violaciones.

Use, knowledge, and attitudes toward evidence-based practice among nursing staff.

BACKGROUND: Little information is known about the use, knowledge, and attitudes toward evidence-based practice (EBP) among nurses in a large academic hospital. This cross-sectional, descriptive study examined the knowledge, attitudes, and use of EBP by nurses at a large academic, Magnet(®)-designated medical center. METHODS: Data were collected from 593 nurses who completed the Clinical Effectiveness and Evidence Based Practice Questionnaire between November 2011 and March 2012. Statistical analyses included correlations and multivariate analysis of covariance. RESULTS: Most nurses (96%) reported that they were aware that an EBP and Research Council existed. The average scores were highest on the Attitudes subscale, followed by the Knowledge/Skills and Practice subscales. CONCLUSION: Continuing education for nurses makes a difference in nurses' attitudes, knowledge, and use of EBP in practice. Participation in EBP and research educational activities or Council meetings may affect EBP culture in a large academic medical center.

Pediatric endophthalmitis. A 10-year consecutive series.

PURPOSE: To report the etiology, culture results, management techniques, and visual outcomes for pediatric endophthalmitis at Emory University for the past 10 years. METHODS: Retrospective chart review based on a computer generated database of all cases of endophthalmitis occurring in patients younger than 18 years from 1995 to 2005. RESULTS: Sixteen cases of pediatric endophthalmitis were identified. Ocular trauma and previous ocular surgery were the most common etiologies, accounting for 44% and 38% of cases respectively. Nine cases (56%) had positive vitreous cultures. Sixty-six percent of cases with positive vitreous cultures involved Gram-positive species. All patients were treated with intravitreal antibiotics. Primary vitrectomy was performed in nine cases. Subsequent to the intraocular infection, 6 eyes (38%) developed retinal detachments. At last follow-up, visual acuity of 20/400 or better was obtained in only 6 eyes (38%). CONCLUSIONS: Pediatric endophthalmitis in this series most commonly resulted from penetrating ocular trauma or from ocular surgery. Retinal detachment commonly occurred as a complication. Despite aggressive management with antibiotics and in many cases vitrectomy, visual outcomes were poor in this series.

Ultrasound biomicroscopy (UBM) characteristics of scleral tunnels created with suture needles commonly used during strabismus surgery.

PURPOSE: To enhance the safety and efficacy of surgical treatment of strabismus, we sought to measure and determine the ultrasound biomicroscopy (UBM) profile of scleral tunnels created with needles commonly used during strabismus surgery, to determine which needles are less likely to create the complication of scleral perforation. METHODS: Adult cadaver eyes were secured in a styrofoam head. Intraocular pressure was maintained between 15 and 21 mm Hg. Then S14, S24, S28 and TG100 needles were used to create scleral tunnels simulating those created during routine strabismus surgery. Ten scleral tunnels were created with each needle type at 3 different sites on the globe, for a total of 120 passes. The thickness of the sclera and the maximum depth and length of each scleral tunnel were measured using UBM. RESULTS: The mean tunnel depth below the scleral surface (+/- SD) was 0.43 +/-0.11 mm, 0.37 +/-0.09, 0.40 +/-0.08 and 0.34 +/-0.07 mm, for the S14, S24, S28 and TG100 needles, respectively (P=0.002, One way ANOVA). For both the S14 and S28 needles, there was a "statistically significant" P 0.05) linear trend of an increase in the depth of the pass as the length of the pass increased (P=0.01 for the S14 and P=0.02 for the S28 {Pearson Correlation 2 tailed test}). A similar trend was found with the S24 needle but the trend was not "statistically significant" (P=0.35). No such trend was found with the TG100 needle. CONCLUSIONS: Needle design had a definite impact on the characteristics of scleral tunnels created to simulate those made during strabismus surgery and may influence needle selection by the surgeon for different or various surgical circumstances, but the differences were not such as to predicate for or against the general use of any of these four needles for strabismus surgery.

The prevalence of pain and pain interference in a general population of older adults: cross-sectional findings from the North Staffordshire Osteoarthritis Project (NorStOP).

Although pain is experienced at all ages, there is uncertainty about the pattern of its occurrence in older people. We have investigated the prevalence of three aspects of self-reported pain-occurrence of any recent pain, number and location of pain sites, and interference with daily life-to determine their association with age in older people. A cross-sectional postal survey of all adults aged 50 years and over registered with three general practices (n = 11230) in North Staffordshire using self-complete questionnaires was conducted. Respondents' gender, age, employment status, socio-economic classification, and general health status were gathered to characterise the population under study. The location of any recent pain (past 4 weeks) was recorded on a full-body manikin and pain interference was based on a single question. Completed questionnaires were received from 7878 respondents (adjusted response of 71.3%). The 4-week prevalence of any pain was 72.4%; similar across 10-year age-groups, and higher in females than males. In those with pain the median number of painful areas (from 44) was 6, and 12.5% of the responding population were classified as having widespread pain, both figures similar across age-groups. Most regional pains showed a decline in prevalence in the older age-groups, the exceptions being the lower limb regions (hip, knee, foot). Pain that interfered with daily activities was reported by 3002 (38.1%) respondents overall. There was a clear age-related rise in this prevalence with age up to and including the oldest group. Within each regional pain subgroup, the proportion of people who also reported pain interference rose with age. Our study has provided evidence that increasing age in the elderly population is not associated with any change in the overall prevalence of pain, although, as previous studies have suggested, the pattern of pain prevalence in different body regions does change with age. More importantly the extent to which pain interferes with everyday life increases incrementally with age up to the oldest age-group in the community-dwelling general population.

Keratoconus-like topographic changes in keratoconjunctivitis sicca.

PURPOSE: To describe a case of inferior corneal steepening with a keratoconus-like pattern in a patient with nocturnal lagophthalmos and aqueous tear deficiency (ATD). METHODS: Axial curvature mapping was performed with the Tomey TMS-2N videokeratoscopy instrument. Keratoconus-like topography patterns were identified with the Klyce Software package of this instrument. RESULTS: Axial videokeratoscopy showed asymmetric inferior corneal steepening in the right eye. The surface regularity index (SRI), surface asymmetry index (SAI), and the simulated keratometric cylinder change (CYL) were 1.05, 5.05, and 0.75, respectively. The keratoconus screening indices were as follows for the right eye: Klyce/Maeda index of 95% similarity (clinical keratoconus interpreted) and Smolek/Klyce index of 23.09% severity (keratoconus suspect interpreted). Ultrasound pachymetric mapping showed a normal central corneal thickness. We found a similar topographic pattern of keratoconus in seven eyes of 74 dry eye patients who were previously evaluated with the Tomey TMS-2N. CONCLUSION: Chronic ocular desiccation and aqueous tear deficiency can produce inferior corneal steepening and high astigmatism resembling keratoconus.