Pyrimidine depletion enhances targeted and immune therapy combinations in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a fatal disease characterized by the accumulation of undifferentiated myeloblasts, and agents that promote differentiation have been effective in this disease but are not curative. Dihydroorotate dehydrogenase inhibitors (DHODHi) have the ability to promote AML differentiation and target aberrant malignant myelopoiesis. We introduce HOSU-53, a DHODHi with significant monotherapy activity, which is further enhanced when combined with other standard-of-care therapeutics. We further discovered that DHODHi modulated surface expression of CD38 and CD47, prompting the evaluation of HOSU-53 combined with anti-CD38 and anti-CD47 therapies, where we identified a compelling curative potential in an aggressive AML model with CD47 targeting. Finally, we explored using plasma dihydroorotate (DHO) levels to monitor HOSU-53 safety and found that the level of DHO accumulation could predict HOSU-53 intolerability, suggesting the clinical use of plasma DHO to determine safe DHODHi doses. Collectively, our data support the clinical translation of HOSU-53 in AML, particularly to augment immune therapies. Potent DHODHi to date have been limited by their therapeutic index; however, we introduce pharmacodynamic monitoring to predict tolerability while preserving antitumor activity. We additionally suggest that DHODHi is effective at lower doses with select immune therapies, widening the therapeutic index.

Inhibition of Enhancer of Zeste Homolog 2 Induces Blast Differentiation, Impairs Engraftment and Prolongs Survival in Murine Models of Acute Myeloid Leukemia.

BACKGROUND: Acute myeloid leukemia (AML) is the malignant proliferation of immature myeloid cells characterized by a block in differentiation. As such, novel therapeutic strategies to promote the differentiation of immature myeloid cells have been successful in AML, although these agents are targeted to a specific mutation that is only present in a subset of AML patients. In the current study, we show that targeting the epigenetic modifier enhancer of zeste homolog 2 (EZH2) can induce the differentiation of immature blast cells into a more mature myeloid phenotype and promote survival in AML murine models. METHODS: The EZH2 inhibitor EPZ011989 (EPZ) was studied in AML cell lines, primary in AML cells and normal CD34+ stem cells. A pharmacodynamic assessment of H3K27me3; studies of differentiation, cell growth, and colony formation; and in vivo therapeutic studies including the influence on primary AML cell engraftment were also conducted. RESULTS: EPZ inhibited H3K27me3 in AML cell lines and primary AML samples in vitro. EZH2 inhibition reduced colony formation in multiple AML cell lines and primary AML samples, while exhibiting no effect on colony formation in normal CD34+ stem cells. In AML cells, EPZ promoted phenotypic evidence of differentiation. Finally, the pretreatment of primary AML cells with EPZ significantly delayed engraftment and prolonged the overall survival when engrafted into immunodeficient mice. CONCLUSIONS: Despite evidence that EZH2 silencing in MDS/MPN can promote AML pathogenesis, our data demonstrate that the therapeutic inhibition of EZH2 in established AML has the potential to improve survival.

Differences in health care provider screening for tobacco use among youth in the United States: The National Youth Tobacco Survey, 2021.

OBJECTIVE: Health care providers (HCP) are encouraged to screen youth for tobacco product use as a key step in preventing such use and associated health outcomes. However, recent data examining differences in HCP tobacco screening by sociodemographic characteristics and tobacco use is scant. METHODS: Data from the 2021 National Youth Tobacco Survey (N = 14,685) were analyzed. Three types of HCP screening were examined: no screening, any e-cigarette use (e-cigarette only, e-cigarette and other tobacco), and non-e-cigarette tobacco product use. Differences by HCP screening were examined using multinomial logistic regression adjusted for age, sex, gender identity, sexual orientation, race/ethnicity, and tobacco use (non-current, sole, dual/poly). RESULTS: Among the sample, 42.8% were screened for any tobacco use, with 30.6% screened for any e-cigarette use and 12.2% for non-e-cigarette tobacco product use only. Youth who were older (vs. younger) (OR = 5.98, 95% CI: 4.78-7.49) and gay/lesbian (vs. heterosexual) (OR = 1.47, 95% CI: 1.02-2.12) were more likely to be screened for e-cigarette use. Youth who were non-Hispanic Black (vs. non-Hispanic White) were less likely to be screened for e-cigarette use (OR = 0.53, 95% CI: 0.42-0.67) and more likely to be screened for non-e-cigarette tobacco use (OR = 1.34, 95% CI: 1.10-1.63). Current sole tobacco use (vs. non-current use) and dual/poly tobacco use (vs. non-current use) increased the likelihood for HCP screening for e-cigarette use. CONCLUSIONS: The majority of U.S. youth continue to not be screened for tobacco use by their HCP. Evidence of disparities in tobacco use screening suggest the need for policies and training that promote equity in screening.

Differences in health care provider advice on abstaining from tobacco use: Findings from the 2020 National Youth Tobacco Survey.

BACKGROUND: Health care providers' (HCP) advice on tobacco prevention and cessation is critical in addressing the tobacco use epidemic among adolescents. However, examination of whether receiving advice from HCPs differs by adolescent sociodemographic characteristics and tobacco use is limited. METHODS: HCP advice to abstain from using tobacco was examined using the 2020 National Youth Tobacco Survey (N = 12,483). Sociodemographic (age, sex, sexual orientation, race/ethnicity, and region) and tobacco use (non-current, sole, dual/poly) differences by HCP advice were evaluated using adjusted logistic regression models. RESULTS: Among the sample, 37.26% of adolescents received HCP advice to abstain from using tobacco products, and 31.35% received HCP advice to abstain from using e-cigarettes specifically. Adolescents who were non-Hispanic Black (vs. non-Hispanic White) were more likely not to receive HCP advice to abstain from all tobacco products (OR = 1.31, 95% CI: 1.08-1.59). Adolescents who were non-Hispanic Black (vs. non-Hispanic White) (OR = 1.41, 95% CI: 1.16-1.73) or sexual minority (vs. heterosexual) (OR = 1.16, 95% CI: 1.02-1.33) were more likely not to receive HCP advice to abstain from e-cigarettes. Adolescents who were aged 16-18 (vs. aged 9-12) (OR = 0.68, 95% CI: 0.56-0.83) or currently use dual/poly tobacco products (vs. adolescents who do not currently use tobacco) (OR = 0.56, 95% CI: 0.43-0.74) were more likely to receive HCP advice to abstain from using e-cigarettes). CONCLUSIONS: Many U.S. adolescents do not receive HCP advice to abstain from using tobacco. HCPs should increase tobacco prevention and cessation advice across adolescent groups, particularly racial/ethnic and sexual minorities. HCP training and public health policies that improve delivery of e-cigarette advice to adolescents are essential.

Harm perceptions of secondhand e-cigarette aerosol among youth in the United States.

E-cigarette use among youth remains a public health concern. Although extant literature has examined the perceived harms of cigarette use and secondhand smoke, perceptions of harms associated with secondhand e-cigarette aerosol (SHA) are not well understood. Therefore, we used data from the 2020 U.S. National Youth Tobacco Survey (n = 13,292) in which participants indicated whether SHA caused no harm, little harm, some harm, or a lot of harm. We dichotomized SHA harm perceptions as harmless vs harmful. We included sociodemographics (i.e., age, sex, race/ethnicity, sexual orientation, urbanicity), e-cigarette use characteristics, and SHA exposure as covariates and estimated associations between SHA harm perceptions and each covariate using adjusted logistic regression. Most youth perceived SHA as harmful (87.9 %) compared to harmless (12.1 %). Older youth (vs younger youth) had higher odds of perceiving SHA as harmless, whereas male (vs female) youth had 49 % higher odds (95 % CI: 1.29-1.72) of perceiving SHA as harmless. As the number of days of e-cigarette use in the past 30 days increased (vs non-users), odds of perceiving SHA as harmless increased. Youth exposed to SHA (vs no exposure) in the past 30 days had 35 % higher odds of perceiving SHA as harmless (95 % CI: 1.16-1.57). To conclude, youth SHA harm perceptions varied overall and by sociodemographic characteristics, e-cigarette use, and SHA exposure. Educational campaigns to inform youth of the health risks associated with e-cigarettes and SHA are needed to reduce overall nicotine intake and disparities in nicotine exposure.

Workplace Culture and Biomarkers of Health Risk.

Workplace culture has been studied for impact on health risk; however, connections with robust biologic markers of health remain to be established. We examined associations between the work environment and urinary levels of catecholamines and their metabolites as biomarkers of sympathetic nervous system activity, indicative of stress. We recruited participants (n = 219; 2018-2019) from a cardiovascular risk cohort to investigate workplace culture, well-being, and stress. Participants completed seven questionnaires. Urine samples were used to measure catecholamines and their metabolites by LC/MS/MS. Pearson correlation and linear regression models were used after adjusting for demographics and creatinine. Participants reporting higher well-being had lower urinary levels of dopamine, serotonin, and 3-methoxytyramine. Participants reporting a more engaged and more positive workplace had lower levels of dopamine and 3-methoxytyramine. Reported workplace isolation was correlated with higher levels of dopamine and 3-methoxytyramine. Given correlations between catecholamines, we used 3-methoxytyramine for linear regression. In fully adjusted models, in environments with a more positive culture, levels of 3-methoxytyramine remained lower (ß = -0.065 ± 0.025, p = 0.01) and indicated a positive association between workplace isolation and 3-methoxytyramine (ß = 0.064 ± 0.030, p = 0.04). These findings are consistent with an important relationship between workplace environment and sympathetic nervous system activity.

Tobacco Use Status and Temptation to Try E-Cigarettes among a Sample of Appalachian Youth.

E-cigarettes are commonly used tobacco products among youth populations, including Appalachian youth. However, knowledge of the extent to which tobacco use status relates to temptation to try e-cigarettes is limited. Data from the Youth Appalachian Tobacco Study (n = 1047) were used. Temptation to try e-cigarettes was derived from a 12-item situational inventory. Tobacco use status was defined as never, ever non-e-cigarette, and ever e-cigarette use. A factorial ANOVA was used to estimate the adjusted association between tobacco use status and the e-cigarette use temptation scale. Two-way interaction terms between tobacco use status and gender, and tobacco use status and race/ethnicity, were plotted to depict effect modification. Approximately 10% of youth were ever non-e-cigarette users and 24% were ever e-cigarette users. Never and ever non-e-cigarette user middle schoolers had higher temptation to try e-cigarettes than their high school counterparts. The same relationship was found among never and ever e-cigarette users living in households with tobacco users. The ANOVA results suggest a positive, monotonic relationship between tobacco use status and temptation to try e-cigarettes, and that the adjusted group means differ by gender and race/ethnicity. The findings can inform tobacco prevention interventions for youth at higher risk for e-cigarette use, especially youth who have not yet tried e-cigarettes.

Microbiome data analysis with applications to pre-clinical studies using QIIME2: Statistical considerations.

Diversity analysis and taxonomic profiles can be generated from marker-gene sequence data with the help of many available computational tools. The Quantitative Insights into Microbial Ecology Version 2 (QIIME2) has been widely used for 16S rRNA data analysis. While many articles have demonstrated the use of QIIME2 with suitable datasets, the application to pre-clinical data has rarely been talked about. The issues involved in the pre-clinical data include the low-quality score and small sample size that should be addressed properly during analysis. In addition, there are few articles that discuss the detailed statistical methods behind those alpha and beta diversity significance tests that researchers are eager to find. Running the program without knowing the logic behind it is extremely risky. In this article, we first provide a guideline for analyzing 16S rRNA data using QIIME2. Then we will talk about issues in pre-clinical data, and how they could impact the outcome. Finally, we provide brief explanations of statistical methods such as group significance tests and sample size calculation.

P027 Brain Fog in Patients With Inflammatory Bowel Disease, and Association With Use of Probiotics.

BACKGROUND: Brain fog has been minimally studied in patients with inflammatory bowel disease (IBD). IBD patients frequently consume probiotics, whether sanctioned by a physician or not. However, probiotic consumption in itself has been shown to increase the incidence of brain fog. We aimed to study the association between brain fog in IBD patients with or without probiotic use. METHODS: We conducted a cross-sectional study among patients visiting a busy IBD clinic. Patients aged >18 with a biopsy-proven diagnosis of inflammatory bowel disease, without pre-existing psychiatric illness or current use of opioid medications were included. They were divided into 2 groups: those using probiotics and those who did not. Patients were given a questionnaire that included details about symptoms of brain fog. Groups were analyzed by chi-square test for differences in baseline demographics, and Mann Whitney U test to compare outcomes between groups. A p-value < 0.05 was considered statistically significant. RESULTS: Of the 66 patients included (mean age 44±2 years), 35 (53%) were female and 59 (89.4%) were Caucasian. Among these patients, 31.8% (n = 21) took probiotics as dietary supplements with the majority (67%, n = 14) taking probiotics for over a year. Overall, there was a trend for an association between probiotic use and brain fog in all patients (p = 0.080) but no statistical significance was attained. However, brain fog was significantly associated with probiotic use among Caucasian patients (p = 0.044). Furthermore, there was a statistically significant association between brain fog and male patients using probiotics (p = 0.004). Duration of probiotic use was also associated with brain fog (p = 0.038). CONCLUSION: Consumption of probiotics was independently associated with brain fog in men, as well as Caucasian patients with IBD respectively. Given the high prevalence of probiotic use in IBD patients, prospective studies are warranted to examine the causal relationship between probiotics and IBD-associated brain fog to guide prescription of probiotic supplements for IBD.

Associations between Appalachian youth tobacco consumption and communication channel use.

INTRODUCTION: Youth tobacco use rates in Appalachia exceed the US national average, and e-cigarette use has increased. Thus, further research is needed to understand how these youth receive and share product information. METHODS: Middle and high school students in rural Appalachia were surveyed (N=1103). The primary outcome, tobacco use, was categorized as: never users, cigarette-only users, smokeless-only users, e-cigarette-only users, and polytobacco users. Associations between receiving or sharing conventional tobacco-related or e-cigarette-related information via specific communication channels and tobacco use were assessed. RESULTS: Participants who received tobacco information from friends and family (FF) (OR=1.87; 95% CI: 1.35-2.57), public displays (PUB) (OR=1.49; 95% CI: 1.09-2.05), and digital media (DIG) (OR=1.95; 95% CI: 1.41-2.70) and e-cigarette information via the same communication channels, FF (OR=2.65; 95% CI: 1.93-3.65), PUB (OR=1.62; 95% CI: 1.17-2.26), and DIG (OR=2.24; 95% CI: 1.61-3.12), had greater odds of being polytobacco users, compared to never users. Participants who received e-cigarette-related information from FF (OR=2.42; 95% CI: 1.42-4.13) and PUB (OR=2.13; 95% CI: 1.25-3.65) had greater odds of being e-cigarette-only users compared to never users. Participants who shared e-cigarette-related information with FF had greater odds of being e-cigarette-only users (OR=3.16; 95% CI: 1.80-5.58) and polytobacco users (OR=4.48; 95% CI: 3.16-6.35) compared to never users. CONCLUSIONS: Receiving and sharing tobacco-related and e-cigarette-related information via multiple communication channels is associated with e-cigarette and polytobacco use among Appalachian youth. Several channels may need to be utilized in health campaigns to influence youth.

Tobacco use and harm perceptions among Appalachian youth.

Appalachian youth tobacco use rates exceed the national average. Additional inquiry is needed to better understand youth product perceptions and use patterns. This study examined tobacco harm perceptions and their relationship with tobacco use among Appalachian youth. From 2014 to 2016, a survey of Appalachian middle and high school students (N = 1,136) was conducted. Tobacco harm perceptions were assessed by tobacco use status, categorized as never user, cigarette only user, smokeless only user, e-cigarette only user, or polytobacco user. Descriptive characteristics were compared by tobacco use status and harm perceptions. Adjusted multivariable logistic regression models assessed the relationship between tobacco use status and harm perceptions. Over one-third of participants were tobacco users (34.6%; 4.7% cigarette only users, 3.3% smokeless only users, 5.6% e-cigarette only users, and 21.0% polytobacco users). Approximately half agreed that e-cigarettes cause health problems (54.4%), and 64.7% agreed that e-cigarettes are addictive. Most participants (83.4-92.3%) agreed that smoking and smokeless tobacco cause health problems and are addictive. Tobacco users more often disagreed that tobacco products cause health problems than did never users. Compared to never users, e-cigarette only users were more likely to disagree that smoking (AOR: 2.99, 95% CI: 1.30-6.90) and e-cigarettes cause health problems (AOR: 2.79, 95% CI: 1.64-4.75) and that e-cigarettes cause addiction (AOR: 2.48, 95% CI: 1.48-4.16). Most youth were aware of health dangers associated with smoking, but perceptions were split on whether e-cigarettes were associated with health problems or addiction. The findings indicate the need for additional youth tobacco use prevention efforts.

Pyoderma Gangrenosum in Inflammatory Bowel Disease: A Systematic Review and Meta-Analysis.

BACKGROUND: Pyoderma gangrenosum (PG) is an uncommon but severe extra-intestinal manifestation (EIM) of inflammatory bowel disease (IBD). The incidence and risk factors for PG are disputed. AIMS: To assess the incidence of PG and identify factors associated with PG in IBD patients. METHODS: A search of electronic databases (Ovid and PubMed) was conducted between 1966 and 2019. Studies that calculated the incidence of PG in IBD patient cohorts were included. Patient demographics, IBD subtype, and EIM presence were recorded. A review of our institutional database of 1057 IBD patients was conducted. A multivariate regression model and meta-analysis were conducted to identify risk factors for PG. A random effects model was used to combine the data of included studies. RESULTS: Fourteen studies were included in addition to 1057 IBD patients and 26 PG cases from the Louisville cohort. In total, there were 379 cases of PG in the cumulative cohort of 61,695 IBD patients. The PG incidence in individual studies ranged from 0.4 to 2.6%. In the institutional cohort, ocular EIMs and a permanent stoma were significant risk factors for PG. In the meta-analysis, PG was associated with female gender (RR = 1.328, 95% CI 1.161-1.520), Crohn's disease (RR = 1.193, 95% CI 1.001-1.422), erythema nodosum (RR = 9.281, 95% CI 6.081-14.164), and ocular EIM (RR = 4.55, 95% CI 3.04-6.81). There was study heterogeneity when assessing IBD subtype, ocular, and joint EIMs. CONCLUSIONS: There are conflicting data on the incidence and risk factors for PG. This meta-analysis confirms an association between PG and female gender, Crohn's disease, erythema nodosum, and ocular EIM that have been described in smaller studies.

A Comparative Evaluation of Conventional and Staircase Modification of the Intraoral Mandibular Vestibular Incision Approach in Symphysis and Parasymphysis Fractures.

AIM: The aim of this study was therefore to evaluate the conventional intraoral mandibular vestibular incision approach in symphysis and parasymphysis fractures and compare prognosis of the incision site, fracture healing, and associated complications with the staircase modification of the intraoral mandibular vestibular incision approach for symphysis and parasymphysis fractures. MATERIALS AND METHODS: This study was conducted on 34 healthy individual of age 18-60 years, reporting to the department with a history of trauma having mandibular symphysis or parasymphysis fractures and underwent open reduction and internal fixation under either local or general anesthesia. The treated patients were prospectively followed and examined for the postoperative complications such as pain, swelling, infection, dehiscence, sensory disturbances, and nonunion/malunion of the fracture site. Patients were followed up at the intervals of 2nd postoperative day, 1 week, 2 weeks, and 6 weeks postoperatively and were evaluated for any of the above complications. RESULTS: There was no statistically significant difference in the assessment parameters between the conventional intraoral mandibular vestibular approach and the staircase modification of the same. CONCLUSION: The conventional method and the staircase modification of the intraoral mandibular vestibular approach have similar treatment outcomes in terms of osteosynthesis and soft tissue healing but the staircase modification fairs better in terms of healing till the 6th week. CLINICAL SIGNIFICANCE: This study contributes to the understanding of the comparatively treatment outcomes of the conventional and staircase modification of the intraoral mandibular vestibular approach with respect to postoperative complications such as pain, swelling, infection, dehiscence, sensory disturbances, and nonunion/malunion of the fracture site, which may influence the choice by the dental surgeon.

Mitochondrial matrix metalloproteinase activation decreases myocyte contractility in hyperhomocysteinemia.

Cardiomyocyte N-methyl-d-aspartate receptor-1 (NMDA-R1) activation induces mitochondrial dysfunction. Matrix metalloproteinase protease (MMP) induction is a negative regulator of mitochondrial function. Elevated levels of homocysteine [hyperhomocysteinemia (HHCY)] activate latent MMPs and causes myocardial contractile abnormalities. HHCY is associated with mitochondrial dysfunction. We tested the hypothesis that HHCY activates myocyte mitochondrial MMP (mtMMP), induces mitochondrial permeability transition (MPT), and causes contractile dysfunction by agonizing NMDA-R1. The C57BL/6J mice were administered homocystinemia (1.8 g/l) in drinking water to induce HHCY. NMDA-R1 expression was detected by Western blot and confocal microscopy. Localization of MMP-9 in the mitochondria was determined using confocal microscopy. Ultrastructural analysis of the isolated myocyte was determined by electron microscopy. Mitochondrial permeability was measured by a decrease in light absorbance at 540 nm using the spectrophotometer. The effect of MK-801 (NMDA-R1 inhibitor), GM-6001 (MMP inhibitor), and cyclosporine A (MPT inhibitor) on myocyte contractility and calcium transients was evaluated using the IonOptix video edge track detection system and fura 2-AM. Our results demonstrate that HHCY activated the mtMMP-9 and caused MPT by agonizing NMDA-R1. A significant decrease in percent cell shortening, maximal rate of contraction (-dL/dt), and maximal rate of relaxation (+dL/dt) was observed in HHCY. The decay of calcium transient amplitude was faster in the wild type compared with HHCY. Furthermore, the HHCY-induced decrease in percent cell shortening, -dL/dt, and +dL/dt was attenuated in the mice treated with MK-801, GM-6001, and cyclosporin A. We conclude that HHCY activates mtMMP-9 and induces MPT, leading to myocyte mechanical dysfunction by agonizing NMDA-R1.