p53-Bad* Fusion Gene Therapy Induces Apoptosis In Vitro and Reduces Zebrafish Tumor Burden in Hepatocellular Carcinoma.

With few curative treatments and a global yearly death rate of over 800,000, hepatocellular carcinoma (HCC) desperately needs new therapies. Although wild-type p53 gene therapy has been shown to be safe in HCC patients, it has not shown enough efficacy to merit approval. This work aims to show how p53 can be re-engineered through fusion to the pro-apoptotic BH3 protein Bcl-2 antagonist of cell death (Bad) to improve anti-HCC activity and potentially lead to a novel HCC therapeutic, p53-Bad*. p53-Bad* is a fusion of p53 and Bad, with two mutations, S112A and S136A. We determined mitochondrial localization of p53-Bad* in liver cancer cell lines with varying p53 mutation statuses via fluorescence microscopy. We defined the apoptotic activity of p53-Bad* in four liver cancer cell lines using flow cytometry. To determine the effects of p53-Bad* in vivo, we generated and analyzed transgenic zebrafish expressing hepatocyte-specific p53-Bad*. p53-Bad* localized to the mitochondria regardless of the p53 mutation status and demonstrated superior apoptotic activity over WT p53 in early, middle, and late apoptosis assays. Tumor burden in zebrafish HCC was reduced by p53-Bad* as measured by the liver-to-body mass ratio and histopathology. p53-Bad* induced significant apoptosis in zebrafish HCC as measured by TUNEL staining but did not induce apoptosis in non-HCC fish. p53-Bad* can induce apoptosis in a panel of liver cancer cell lines with varying p53 mutation statuses and induce apoptosis/reduce HCC tumor burden in vivo in zebrafish. p53-Bad* warrants further investigation as a potential new HCC therapeutic.

When Do Anisotropic Magnetic Susceptibilities Lead to Large NMR Shifts? Exploring Particle Shape Effects in the Battery Electrode Material LiFePO4.

Materials used as electrodes in energy storage devices have been extensively studied with solid-state NMR spectroscopy. Due to the almost ubiquitous presence of transition metals, these systems are also often magnetic. While it is well known that the presence of anisotropic bulk magnetic susceptibility (ABMS) leads to broadening of resonances under magic angle spinning, we show that for monodisperse and nonspherical particle morphologies the ABMS can also lead to considerable shifts, which vary substantially as a function of particle shape. This, on one hand, complicates the interpretation of the NMR spectrum and means that different samples of the same nominal material may no longer give rise to the same measured shift. On the other hand, the ABMS shift provides a mechanism with which to derive the particle shape from the NMR spectrum. In this work, we present a methodology to model the ABMS shift and relate it to the shape of the studied particles. The approach is tested on the 7Li NMR spectra of single crystals and powders of LiFePO4. The results show that the ABMS shift can be a major contribution to the total NMR shift in systems with large magnetic anisotropies and small hyperfine shifts, 7Li shifts for typical LiFePO4 morphologies varying by as much as 100 ppm. The results are generalized to demonstrate that the approach can be used as a means with which to probe the aspect ratio of particles. The work has implications for the analysis of NMR spectra of all materials with anisotropic magnetic susceptibilities, including diamagnetic materials such as graphite.

Crystallites of magnetic charges in artificial spin ice.

Artificial spin ice is a class of lithographically created arrays of interacting ferromagnetic nanometre-scale islands. It was introduced to investigate many-body phenomena related to frustration and disorder in a material that could be tailored to precise specifications and imaged directly. Because of the large magnetic energy scales of these nanoscale islands, it has so far been impossible to thermally anneal artificial spin ice into desired thermodynamic ensembles; nearly all studies of artificial spin ice have either treated it as a granular material activated by alternating fields or focused on the as-grown state of the arrays. This limitation has prevented experimental investigation of novel phases that can emerge from the nominal ground states of frustrated lattices. For example, artificial kagome spin ice, in which the islands are arranged on the edges of a hexagonal net, is predicted to support states with monopolar charge order at entropies below that of the previously observed pseudo-ice manifold. Here we demonstrate a method for thermalizing artificial spin ices with square and kagome lattices by heating above the Curie temperature of the constituent material. In this manner, artificial square spin ice achieves unprecedented thermal ordering of the moments. In artificial kagome spin ice, we observe incipient crystallization of the magnetic charges embedded in pseudo-ice, with crystallites of magnetic charges whose size can be controlled by tuning the lattice constant. We find excellent agreement between experimental data and Monte Carlo simulations of emergent charge-charge interactions.

GROW: Green Organic Vegetable Gardens to Promote Older Adult Wellness: a Feasibility Study.

Protecting older adult's cognitive health is a public health priority. Wellness behaviors within 6 domains have demonstrated effectiveness in protecting older adult's cognitive abilities. Interventions targeted to low-income older adults are needed because these populations experience greater social and physical health disparities compared to adults in higher socioeconomic statuses. This study examined the feasibility of engaging independent, community-dwelling older adults living in low-income senior housing in cultivating raised-bed gardens and reviewed the improvements in cognition and nutrition. Ten participants received ergonomic garden tools, seeds, waist-height garden beds, and weekly garden education. Participants planted vegetables of their choosing and tended to their garden beds for 17 weeks. Cognition and nutrition outcomes were measured before and after the intervention. Adults successfully engaged in the intervention throughout the duration and experienced improvement in cognitive and nutrition outcomes.

Maternal Buprenorphine Dose at Delivery and Its Relationship to Neonatal Outcomes.

BACKGROUND/AIMS: To determine whether there is a dose-response relationship between maternal dose of buprenorphine at delivery and neonatal outcomes. METHODS: This retrospective cohort study of 155 maternal-infant dyads exposed to buprenorphine during pregnancy examines the relationship between maternal dose of buprenorphine at delivery and gestational age, birthweight, method of delivery, Apgar scores at 1 and 5 min, duration of infant hospital stay, peak neonatal abstinence syndrome (NAS) score, duration of NAS and incidence of pharmacologic treatment of NAS. RESULTS: Analyses failed to support any relationship between maternal dose of buprenorphine at delivery and any of the 9 clinical outcomes (all p values >0.093). CONCLUSIONS: This study failed to provide any evidence to support limiting or reducing maternal dose of buprenorphine during pregnancy in order to reduce possible adverse outcomes to the infant. Findings suggest that healthcare providers can focus medication decisions on maternal opioid cravings to reduce the risk of relapse to illicit opioid use rather than out of concern for adverse infant outcomes.

Assessment of trunk muscle density using CT and its association with degenerative disc and facet joint disease of the lumbar spine.

OBJECTIVE: The purpose of this study was (1) to evaluate the association of trunk muscle density assessed by computed tomography (CT) with age, gender, and BMI and (2) to evaluate the association between trunk muscle CT density and degenerative disc and facet joint disease of the lumbar spine. MATERIAL AND METHODS: The study was IRB approved and HIPAA compliant. The study group comprised 100 subjects (mean age 44.4 ± 22.2 years, 51 % male) who underwent CT of the abdomen and pelvis without intravenous contrast. Exclusion criteria included prior abdominal or spine surgery, active malignancy and scoliosis. CTs were reviewed and the attenuation of the rectus abdominis, transverse abdominis, internal and external obliques, psoas, multifidus, longissimus and gluteus maximus were measured bilaterally at consistent levels. Degenerative disc and bilateral facet joint disease were scored using established methods. Univariate analyses were performed using linear regression. Multivariate linear regression was performed to adjust for age, gender and BMI. RESULTS: CT density of each trunk muscle correlated inversely with age (p < 0.001) and BMI (p < 0.001). CT density of each trunk muscle correlated inversely with degenerative disc and facet joint disease in the univariate analyses (p < 0.001); however, only the gluteus maximus and the transverse abdominis remained significant predictors of degenerative disc and facet joint disease respectively in the multivariate analysis. CONCLUSION: Fatty infiltration of trunk musculature increases with age and BMI. Fatty infiltration of the gluteus maximus and transverse abdominis are associated with degenerative disc and facet joint disease, independent of age, gender and BMI.

Characterization of Human Engraftment and Hemophagocytic Lymphohistiocytosis in NSG-SGM3 Neonate Mice Engrafted with Purified CD34+ Hematopoietic Stem Cells.

Immunodeficient mice bearing human immune systems, or "humanized" chimeric mice, are widely used in basic research, along with the preclinical stages of drug development. Nonobese diabetic-severe combined immunodeficiency (NOD-SCID) IL2Rγnull (NSG) mice expressing human stem cell factor, granulocyte-macrophage colony stimulating factor, and interleukin-3 (NSG-SGM3) support robust development of human myeloid cells and T cells but have reduced longevity due to the development of fatal hemophagocytic lymphohistiocytosis (HLH). Here, we describe an optimized protocol for development of human immune chimerism in NSG-SGM3 mice. We demonstrate that efficient human CD45+ reconstitution can be achieved and HLH delayed by engraftment of neonatal NSG-SGM3 with low numbers of human umbilical cord-derived CD34+ hematopoietic stem cells in the absence of preconditioning irradiation.

Are there gender related differences in neonatal abstinence syndrome following exposure to buprenorphine during pregnancy?

AIM: To determine whether infant gender influences the course of neonatal abstinence syndrome (NAS) following exposure to buprenorphine during pregnancy. METHODS: A retrospective cohort study was performed in which maternal and infant data were collected for 46 male and 44 female infants. All infants were born to women enrolled in a buprenorphine treatment program from December 2007 until October 2012. Maternal and infant characteristics and outcomes were compared by infant gender. RESULTS: Male infants had a significantly higher mean peak NAS score (10.04 vs. 7.98, P=0.028) and were more likely to require pharmacologic treatment for NAS (39.1% vs. 11.4%, P=0.005). CONCLUSIONS: These data indicate that, following exposure to buprenorphine during pregnancy, male infants experience a more severe withdrawal syndrome and are more likely to require pharmacologic treatment for NAS.

Wellness in Nursing Education to Promote Resilience and Reduce Burnout: Protocol for a Holistic Multidimensional Wellness Intervention and Longitudinal Research Study Design in Nursing Education.

BACKGROUND: The United States faces a nursing shortage driven by a burnout epidemic among nurses and nursing students. Nursing students are an integral population to fuel the nursing workforce at high risk of burnout and increased rates of perceived stress. OBJECTIVE: The aim of this paper is to describe WellNurse, a holistic, interdisciplinary, multidimensional longitudinal research study that examines evidence-based interventions intended to reduce burnout and increase resilience among graduate and undergraduate nursing students. METHODS: Graduate and undergraduate nursing students matriculated at a large public university in the northeastern United States are eligible to enroll in this ongoing, longitudinal cohort study beginning in March 2021. Participants complete a battery of health measurements twice each semester during the fourth week and the week before final examinations. The measures include the Perceived Stress Scale, the Satisfaction with Life Scale, the Oldenburg Burnout Inventory, the Brief Resilience Scale, and the Pittsburgh Sleep Quality Index. Participants are eligible to enroll in a variety of interventions, including mindfulness-based stress reduction, mindful eating, fitness training, and massage therapy. Those who enroll in specific, targeted interventions complete additional measures designed to target the aim of the intervention. All participants receive a free Fitbit device. Additional environmental changes are being implemented to further promote a culture that supports academic well-being, including recruiting a diverse student population through evidence-based holistic admissions, inclusive teaching design, targeted resilience and stress reduction workshops, and cultural shifts within classrooms and curricula. The study design protocol is registered at Open Science Framework (DOI 10.17605/OSF.IO/NCBPE). RESULTS: The project was funded on January 1, 2022. Data collection started in March 2022. A total of 267 participants have been recruited. Results will be published after each semester starting in December 2023. WellNurse evaluation follows the Rapid Cycle Quality Improvement framework to continuously monitor ongoing project processes, activity outcomes, and progress toward reducing burnout and increasing resilience. Rapid Cycle Quality Improvement promotes the ability to alter WellNurse interventions, examine multiple interventions, and test their effectiveness among the nursing education population to identify the most effective interventions. CONCLUSIONS: Academic nursing organizations must address student burnout risk and increase resilience to produce a future workforce that provides high-quality patient care to a diverse population. Findings from WellNurse will support evidence-based implementations for public baccalaureate and master's nursing programs in the United States. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/49020.

Indirect Calorimetry to Measure Metabolic Rate and Energy Expenditure in Psychiatric Populations: A Systematic Review.

Psychiatric and metabolic disorders are highly comorbid and the relationship between these disorders is bidirectional. The mechanisms underlying the association between psychiatric and metabolic disorders are presently unclear, which warrants investigation into the dynamics of the interplay between metabolism, substrate utilization, and energy expenditure in psychiatric populations, and how these constructs compare to those in healthy controls. Indirect calorimetry (IC) methods are a reliable, minimally invasive means for assessing metabolic rate and substrate utilization in humans. This review synthesizes the extant literature on the use of IC on resting metabolism in psychiatric populations to investigate the interaction between psychiatric and metabolic functioning. Consistently, resting energy expenditures and/or substrate utilization values were significantly different between psychiatric and healthy populations in the studies contained in this review. Furthermore, resting energy expenditure values were systematically overestimated when derived from predictive equations, compared to when measured by IC, in psychiatric populations. High heterogeneity between study populations (e.g., differing diagnoses and drug regimens) and methodologies (e.g., differing posture, time of day, and fasting status at measurement) impeded the synthesis of results. Standardized IC protocols would benefit this line of research by enabling meta-analyses, revealing trends within and between different psychiatric disorders.

Potential physiological responses contributing to the ergogenic effects of acute ischemic preconditioning during exercise: A narrative review.

Ischemic preconditioning (IPC) has been reported to augment exercise performance, but there is considerable heterogeneity in the magnitude and frequency of performance improvements. Despite a burgeoning interest in IPC as an ergogenic aid, much is still unknown about the physiological mechanisms that mediate the observed performance enhancing effects. This narrative review collates those physiological responses to IPC reported in the IPC literature and discusses how these responses may contribute to the ergogenic effects of IPC. Specifically, this review discusses documented central and peripheral cardiovascular responses, as well as selected metabolic, neurological, and perceptual effects of IPC that have been reported in the literature.

Peripartum and Postpartum Analgesia and Pain in Women Prescribed Buprenorphine for Opioid Use Disorder Who Deliver by Cesarean Section.

Aim: Little is known about whether pain can be effectively managed in pregnant women with opioid use disorder (OUD) during delivery hospitalization, particularly those undergoing surgery and taking buprenorphine as medication for OUD (MOUD). To address this question, we compared pain scores and opioid analgesic utilization during delivery hospitalization in women taking their pre-hospital dose of buprenorphine who delivered by cesarean section to matched controls. To inform future research efforts, we also began to explore opioid analgesic utilization and pain scores by type of anesthesia as this variable is often not included in related literature. Methods: Retrospective matched cohort study of 46 women prescribed buprenorphine during pregnancy who delivered by cesarean section during a 7-year period. Results: When compared to matched controls, women taking their pre-hospital dose of buprenorphine undergoing cesarean section utilized more opioid analgesics as measured by morphine milligram equivalents (MME) (mean MME first 48 hours 153.0 mg vs 175.1 mg, respectively, P < .01) but had similar pain scores during delivery hospitalization. There was no difference in MME utilization by maternal dose of buprenorphine though sample sizes were small. Women on buprenorphine who received spinal anesthesia with morphine had mean pain scores that were 1.4 points lower (P = .01) during the first 48 hours than women on buprenorphine receiving other methods of anesthesia. Discussion And Conclusions: Pregnant women taking their pre-hospital dose of buprenorphine throughout their surgical delivery hospitalization were able to achieve pain relief similar to women not on MOUD but had higher MME requirements. Our results add to the emerging body of evidence suggesting that individuals on MOUD can achieve adequate post-surgical pain management without adjusting their pre-hospital dose of buprenorphine. Further research is required to fully understand the optimal buprenorphine dosing regimen during surgical hospitalizations. Our results also provide important preliminary evidence that spinal anesthesia containing opioids can be used effectively in individuals with OUD requiring surgical intervention.

Statistical adjustments for brain size in volumetric neuroimaging studies: some practical implications in methods.

Volumetric magnetic resonance imaging (MRI) brain data provide a valuable tool for detecting structural differences associated with various neurological and psychiatric disorders. Analysis of such data, however, is not always straightforward, and complications can arise when trying to determine which brain structures are "smaller" or "larger" in light of the high degree of individual variability across the population. Several statistical methods for adjusting for individual differences in overall cranial or brain size have been used in the literature, but critical differences exist between them. Using agreement among those methods as an indication of stronger support of a hypothesis is dangerous given that each requires a different set of assumptions be met. Here we examine the theoretical underpinnings of three of these adjustment methods (proportion, residual, and analysis of covariance) and apply them to a volumetric MRI data set. These three methods used for adjusting for brain size are specific cases of a generalized approach which we propose as a recommended modeling strategy. We assess the level of agreement among methods and provide graphical tools to assist researchers in determining how they differ in the types of relationships they can unmask, and provide a useful method by which researchers may tease out important relationships in volumetric MRI data. We conclude with the recommended procedure involving the use of graphical analyses to help uncover potential relationships the ROI volumes may have with head size and give a generalized modeling strategy by which researchers can make such adjustments that include as special cases the three commonly employed methods mentioned above.

Impact of Maine's statewide nutrition policy on high school food environments.

INTRODUCTION: We assessed the effect on the food environments of public high schools of Maine's statewide nutrition policy (Chapter 51), which banned "foods of minimal nutritional value" (FMNV) in public high schools that participated in federally funded meal programs. We documented allowable exceptions to the policy and describe the school food environments. METHODS: We mailed surveys to 89 high school food-service directors to assess availability pre-Chapter 51 and post-Chapter 51 of soda, other sugar-sweetened beverages, and junk food. Frequency data were tabulated pre-Chapter 51 and post-Chapter 51, and Fisher exact test was used to assess significance in changes. We conducted food and beverage inventories at 11 high schools. RESULTS: The survey return rate was 61% (N = 54). Availability of soda in student vending significantly decreased pre-Chapter 51 versus post-Chapter 51 (P = .04). No significant changes were found for other sugar-sweetened beverages and junk food. Exceptions to Chapter 51 were permitted to staff (67%), to the public (86%), and in career and technical education programs (31%). Inventories in a subset of schools found no availability of soda for students, whereas other sugar-sweetened beverages and junk food were widely available in à la carte, vending machines, and school stores. Candy, considered a FMNV, was freely available. Soda advertisement on school grounds was common. CONCLUSION: Student vending choices improved after the implementation of Chapter 51; however, use of FMNV as the policy standard may be limiting, as availability of other sugar-sweetened beverages and junk food was pervasive. School environments were not necessarily supportive of the policy, as advertisement of soda was common and some FMNV were available. Furthermore, local exceptions to Chapter 51 likely reduced the overall effect of the policy.

Methodological Variations Contributing to Heterogenous Ergogenic Responses to Ischemic Preconditioning.

Ischemic preconditioning (IPC) has been repeatedly reported to augment maximal exercise performance over a range of exercise durations and modalities. However, an examination of the relevant literature indicates that the reproducibility and robustness of ergogenic responses to this technique are variable, confounding expectations about the magnitude of its effects. Considerable variability among study methodologies may contribute to the equivocal responses to IPC. This review focuses on the wide range of methodologies used in IPC research, and how such variability likely confounds interpretation of the interactions of IPC and exercise. Several avenues are recommended to improve IPC methodological consistency, which should facilitate a future consensus about optimizing the IPC protocol, including due consideration of factors such as: location of the stimulus, the time between treatment and exercise, individualized tourniquet pressures and standardized tourniquet physical characteristics, and the incorporation of proper placebo treatments into future study designs.

Implications of perinatal buprenorphine exposure on infant head circumference at birth.

Objective: To determine the potential impact of prenatal buprenorphine exposure on head circumference at birth and analyze whether head circumference may be related to maternal buprenorphine dose at delivery, delayed maternal entry into buprenorphine treatment or exposure to a variety of other medications and substances.Methods: A retrospective cohort study was performed of 137 full-term infants exposed to buprenorphine during pregnancy from January 2013 to December 2017. Pearson's correlation was calculated to investigate the potential relationship between head circumference and maternal dose of buprenorphine at delivery. t-tests were conducted to analyze head circumference in relationship to dichotomous variables.Results: Head circumference in infants exposed to buprenorphine during pregnancy was not significantly different from national norms for either male infants (95% CI 28.2-33.5 cm, norm 31.5 cm, and 28.5-34.9 cm, norm 33.1 cm, for the 3rd and 10th percentile, respectively) or female infants (95% CI 28.7-32.8 cm, norm 31.9 cm, and 29.1-34.3, norm 32.8 cm for the 3rd and 10th percentile, respectively). Head circumference was not associated with delayed maternal entry into buprenorphine treatment (t = -1.0715, p = .287) or exposure to psychotropic medications during pregnancy (t = 0.4194, p = .677). There was no relationship between infant head circumference and maternal buprenorphine dose at delivery (r = 0.004, p = .967). Head circumference was not associated with maternal smoking (t = 0.003, p = .998) or exposure to marijuana (t = 0.7277, p = .468), illicit opioids (t = -0.6701, p = .504), illicit amphetamines (t = -0.4062, p = .687) or illicit benzodiazepines (t = -0.6288, p = .535) during pregnancy.Conclusions: Exposure to buprenorphine prenatally does not appear to be associated with reduced head circumference at birth. Head circumference at birth also does not appear related to either maternal buprenorphine dose at delivery or delayed entry into treatment. As previous literature suggests that high dose methadone exposure during pregnancy may be associated with smaller head circumference and that smaller head circumference may be associated with risk of neurocognitive disorders, our results further support the use of buprenorphine as a first line treatment for opioid use disorders during pregnancy.

Mapping the time course of semantic activation in mediated false memory: Immediate classification, naming, and recognition.

We evaluated the time course of persistent automatic spreading activation from a mediated list of indirect associates (e.g., meow, day, and basement) that all converged upon a non-presented critical item (CI; e.g., black). Mediated lists were related to CIs through non-presented mediators (e.g., cat, night, and bottom). Three speeded tasks were used to evaluate the time course of semantic activation of the CI: a continuous semantic classification task (concrete/abstract decisions), a naming task (reading words aloud), or a recognition test (old/new memory decisions). Test lists were presented immediately following the mediated lists, and CIs were presented in the first, third, or eighth positions. The results revealed that in both the classification and naming tasks, CI priming was greatest in the first test position and declined across the remaining test positions. Importantly, priming was statistically reliable in the late test positions, providing evidence for long-term semantic priming (i.e., across positions on immediate tasks). False recognition, however, was stable across test positions. Collectively, these patterns suggest that spreading-activation processes decline, consistent with implicit spreading activation, and these processes may contribute to long-term false recognition.

Human CD141+ dendritic cells (cDC1) are impaired in patients with advanced melanoma but can be targeted to enhance anti-PD-1 in a humanized mouse model.

BACKGROUND: The conventional type 1 dendritic cell subset (cDC1) is indispensable for tumor immune responses and the efficacy of immune checkpoint inhibitor (ICI) therapies in animal models but little is known about the role of the human CD141+ DC cDC1 equivalent in patients with melanoma. METHODS: We developed a flow cytometry assay to quantify and characterize human blood DC subsets in healthy donors and patients with stage 3 and stage 4 metastatic melanoma. To examine whether harnessing CD141+ DCs could improve responses to ICIs in human melanoma, we developed a humanized mouse model by engrafting immunodeficient NSG-SGM3 mice with human CD34+ hematopoietic stem cells (HSCs) from umbilical cord blood followed by transplantation of a human melanoma cell line and treatment with anti-programmed cell death protein-1 (anti-PD-1). RESULTS: Blood CD141+ DC numbers were significantly reduced in patients with stage 4 melanoma compared with healthy controls. Moreover, CD141+ DCs in patients with melanoma were selectively impaired in their ability to upregulate CD83 expression after stimulation with toll-like receptor 3 (TLR3) and TLR7/8 agonists ex vivo. Although DC numbers did not correlate with responses to anti-PD-1 and/or anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) ICIs, their numbers and capacity to upregulate CD83 declined further during treatment in non-responding patients. Treatment with anti-PD-1 was ineffective at controlling tumor growth in humanized mice but efficacy was enhanced by indirectly expanding and activating DCs in vivo with fms-like tyrosine kinase-3 ligand (Flt3L) and a TLR3 agonist. Moreover, intratumoral injections of CD141+ DCs resulted in reduced tumor growth when combined with anti-PD-1 treatment. CONCLUSIONS: These data illustrate quantitative and qualitative impairments in circulating CD141+ DCs in patients with advanced melanoma and that increasing CD141+ DC number and function is an attractive strategy to enhance immunogenicity and response rates to ICIs.