Inhibition of GSK3α,ß rescues cognitive phenotypes in a preclinical mouse model of CTNNB1 syndrome.

CTNNB1 syndrome is a rare monogenetic disorder caused by CTNNB1 de novo pathogenic heterozygous loss-of-function variants that result in cognitive and motor disabilities. Treatment is currently lacking; our study addresses this critical need. CTNNB1 encodes ß-catenin which is essential for normal brain function via its dual roles in cadherin-based synaptic adhesion complexes and canonical Wnt signal transduction. We have generated a Ctnnb1 germline heterozygous mouse line that displays cognitive and motor deficits, resembling key features of CTNNB1 syndrome in humans. Compared with wild-type littermates, Ctnnb1 heterozygous mice also exhibit decreases in brain ß-catenin, ß-catenin association with N-cadherin, Wnt target gene expression, and Na/K ATPases, key regulators of changes in ion gradients during high activity. Consistently, hippocampal neuron functional properties and excitability are altered. Most important, we identify a highly selective inhibitor of glycogen synthase kinase (GSK)3α,ß that significantly normalizes the phenotypes to closely meet wild-type littermate levels. Our data provide new insights into brain molecular and functional changes, and the first evidence for an efficacious treatment with therapeutic potential for individuals with CTNNB1 syndrome.

E-cigarette and combustible cigarette cessation patterns, reasons, and methods among adolescents, young adults, and adults.

Research is limited regarding adolescents' and young adults' (AYA) patterns, methods of, and reasons for cigarette or e-cigarette cessation. Further, while adults may try to use e-cigarettes to quit combustible cigarettes, little is known about how adults then quit e-cigarettes. This study utilizes a national, cross-sectional online survey of 6131 diverse participants aged 13-40 years to examine reasons for quitting e-cigarettes or cigarettes, quit methods, and quit attempt outcomes among AYAs and adults. In our sample, 3137 (51.2%) had ever used an e-cigarette, of whom 2310 (37.7%) were aged 13-24 years and 827 (13.5%) were 25-40 years old; 2387 (38.9%) had ever used a combustible cigarette (1440 [23.5%] were 13-24 years old and 947 [15.4%] were 25-40 years old). Among e-cigarette ever-users, 39.4% of 13-24-year-olds intended to quit in the next 6 months, and 36.9% had a serious plan for quitting in the next 30 days; 25.2% wanted to decrease the amount they used while 34.8% wanted to quit completely. Similar rates were found among e-cigarette ever users aged 25-40 years, as well as past 30-day e-cigarette users, cigarette ever-users, and past 30-day cigarette users across all ages. "Cold turkey" (41.0%) followed by "tried to cut down slowly by vaping/smoking less often or fewer puffs" (25.5%) was the most common quit method among e-cigarette ever-users and cigarette ever-users of all ages. Further study of effective tobacco cessation methods to help both AYAs and adults successfully quit nicotine, whether from cigarettes or e-cigarettes, is urgently needed.

Reconstituting Spore Cortex Peptidoglycan Biosynthesis Reveals a Deacetylase That Catalyzes Transamidation.

Some bacteria survive in nutrient-poor environments and resist killing by antimicrobials by forming spores. The cortex layer of the peptidoglycan cell wall that surrounds mature spores contains a unique modification, muramic-δ-lactam, that is essential for spore germination and outgrowth. Two proteins, the amidase CwlD and the deacetylase PdaA, are required for muramic-δ-lactam synthesis in cells, but their combined ability to generate muramic-δ-lactam has not been directly demonstrated. Here we report an in vitro reconstitution of cortex peptidoglycan biosynthesis, and we show that CwlD and PdaA together are sufficient for muramic-δ-lactam formation. Our method enables characterization of the individual reaction steps, and we show for the first time that PdaA has transamidase activity, catalyzing both the deacetylation of N-acetylmuramic acid and cyclization of the product to form muramic-δ-lactam. This activity is unique among peptidoglycan deacetylases and is notable because it may involve the direct ligation of a carboxylic acid with a primary amine. Our reconstitution products are nearly identical to the cortex peptidoglycan found in spores, and we expect that they will be useful substrates for future studies of enzymes that act on the spore cortex.

Use, marketing, and appeal of oral nicotine products among adolescents, young adults, and adults.

IMPORTANCE: Oral nicotine products such as pouches, lozenges, tablets, gums, and toothpicks are gaining popularity, especially among adolescents and young adults, with increased marketing. OBJECTIVE: To estimate use patterns of oral nicotine products and likelihood of buying and liking products based on marketing, using a large group of adolescents, young adults, and adults. DESIGN, SETTING, AND PARTICIPANTS: A cross-sectional, online survey among U.S. participants (n = 6,131; ages 13-40 years) was conducted in November-December 2021. MAIN OUTCOMES AND MEASURES: Ever, past-30-day, and past-7-day use, behaviors, and flavors of oral nicotine products. Liking marketing and likelihood of buying specific oral nicotine products (Zyn pouches and Lucy gum) from marketing. RESULTS: Our sample included 2,025 (33.0%) ever-users, 1,191 (19.4%) past-30-day users, and 998 (16.3%) past-7-day users of any oral nicotine product. Use patterns by age (in years): ever-users (<21: 816 (22.3%); 21-40: 1,209 (48.9%)); past-30-day users (<21: 458 (12.5%); 21-40: 733 (29.7%)); and past-7-day users (<21: 383 (10.5%); 21-40: 615 (24.9%)). Across products, 10-18% of participants reported using nicotine strength ranging from 6-10 mg. Fruit, sweet/dessert, alcohol, coffee, and mint were the most used flavors. When shown marketing, ever-users liked and were likely to buy Zyn pouches compared to never users, and participants under 21 years felt equally targeted by Lucy and Zyn marketing. Liking Zyn marketing even a little bit compared to not at all increased the likelihood of buying Zyn pouches across age groups. After observing marketing, participants < 21 years were more likely to buy Zyn if they perceived marketing to contain messages about good tasting flavors (AOR 1.43, 1.09-1.87; 0.009) and helping to feel comfortable in social situations (AOR 1.38, 1.02-1.87; 0.033), and were more likely to buy Lucy if they felt it could be used anywhere (AOR 1.57, 1.05-2.33; 0.026). CONCLUSIONS: This study provides a foundation for estimating use, behaviors, flavors, and marketing influence of oral nicotine products in the US and globally. Adolescent and young adult use of oral nicotine products and likelihood of buying products when exposed to marketing highlights the need for expanded tobacco use surveillance, marketing regulations, and counter marketing and educational efforts.

Nicotine Dependence from Different E-Cigarette Devices and Combustible Cigarettes among US Adolescent and Young Adult Users.

E-cigarettes, the most popular tobacco product among adolescents, vary widely in design and nicotine composition; thus, different devices may have different addictive potential. However, few studies examine levels of nicotine dependence across devices among adolescent and young adult (AYA) e-cigarette users. To assess the extent of nicotine dependence among US AYA (ages 13-24) by e-cigarette device type, we conducted a large, national, cross-sectional survey (n = 4351) and used the Hooked on Nicotine Checklist (HONC) to assess levels of nicotine dependence among those who had used disposable, pod-based, and/or mods/other e-cigarette devices in the past 30 days. We also examined HONC scores among those who had used combustible cigarettes in the past 30 days, whether with or without using e-cigarettes. Patterns of nicotine dependence were comparable across those who had used a combustible cigarette and/or e-cigarette in the past 30 days, with 91.4% of combustible cigarette users, 80.7% of disposable e-cigarette users, 83.1% of pod-based e-cigarette users, and 82.5% of mods/other e-cigarette users showing signs of nicotine dependence, as measured by endorsing at least one HONC symptom. This pattern persisted when analyses were restricted to e-cigarette only users, with more than 70% of all e-cigarette only past-30-day users endorsing at least one HONC symptom, across all types of devices. A thorough understanding of the extent and presentation of nicotine dependence among AYA will help researchers, public health officials, and clinicians recognize and manage AYA nicotine dependence.

Use Patterns, Flavors, Brands, and Ingredients of Nonnicotine e-Cigarettes Among Adolescents, Young Adults, and Adults in the United States.

Importance: Nonnicotine e-cigarettes contain chemicals, flavorants, and solvents that have known health harms and/or have not been proven safe for inhalation. Objective: To evaluate nonnicotine e-cigarette use patterns, including common flavors, brands, and ingredients. Design, Setting, and Participants: This cross-sectional study included a convenience sample of US residents aged 13 to 40 years who completed an online survey in November and December 2021. Quota sampling was used for an equal proportion of participants aged 13 to 17 years, 18 to 20 years, and 21 to 40 years, balanced for sex, race, and ethnicity per the latest US Census. Main Outcomes and Measures: Nonnicotine e-cigarette use (ever, past 30- and past 7-day, number of times used, time taken to finish); co-use with nicotine e-cigarettes; age at first try; and flavors, brands, and ingredients used. Results: Overall, 6131 participants (mean [SD] age, 21.9 [6.8] years; range, 13-40 years; 3454 [56.3%] identifying as female) completed the survey (55.1% completion rate). Among all participants, 1590 (25.9%) had ever used a nonnicotine e-cigarette, 1021 (16.7%) used one in the past 30 days, and 760 (12.4%) used one in the past 7 days. By age group, 227 of 1630 participants aged 13 to 17 years (13.9%), 497 of 2033 participants aged 18 to 20 years (24.4%), 399 of 1041 participants aged 21 to 24 years (38.3%), and 467 of 1427 participants aged 25 to 40 years (32.7%) had ever used nonnicotine e-cigarettes. Among 1590 participants who had ever used a nonnicotine e-cigarette, 549 (34.5%) had used one more than 10 times; 1017 (63.9%) finished 1 nonnicotine e-cigarette in less than 1 week. Co-use of nonnicotine with nicotine e-cigarettes was reported by 1155 participants (18.8%), 1363 (22.2%) exclusively used nicotine e-cigarettes, and 431 (7.0%) exclusively used nonnicotine e-cigarettes. Most-used flavors were sweet, dessert, or candy (578 [36.3%]); fruit (532 [33.4%]); and mint or menthol (321 [20.2%]); similar flavor patterns were observed for the top 2 flavors among those who used nonnicotine e-cigarettes in the past 30 days, followed by combinations of coffee, alcohol, flower, plant, and mint or menthol flavors by age group. Participants most reported using tetrahydrocannabinol (587 [36.9%]), cannabidiol (537 [33.7%]), melatonin (438 [27.5%]), caffeine (428 [26.9%]), and essential oils (364 [22.9%]) in their nonnicotine e-cigarettes. Conclusions and Relevance: In this study of adolescents, young adults, and adults, a sizeable proportion reported having used nonnicotine e-cigarettes and co-using them with nicotine e-cigarettes. Surveillance studies should further assess nonnicotine e-cigarette use patterns and regulations, and prevention should be developed to address youth appeal, unsubstantiated health claims, and possible health harms.

Entice With Procedures, Inspire With Primary Care: A Preclerkship Pipeline Course.

BACKGROUND AND OBJECTIVES: The growing demand for primary care clinicians in the United States continues to outstrip their dwindling supply. Many allopathic medical schools, including Stanford University School of Medicine, are not adequately meeting this shortage. We sought to develop a preclerkship elective to increase the visibility and desirability of primary care at our institution. METHODS: A novel 9-week preclerkship elective titled "Primary Care Defined: Perspectives and Procedures," was designed as a series of procedural workshops followed by interactive sessions with local primary care clinicians. A total of 36 medical and physician assistant students were enrolled. We administered a questionnaire pre- and postcourse to evaluate the impact of the elective on learner interest and attitudes toward primary care. RESULTS: Twenty-four enrolled and 10 nonenrolled learners completed the questionnaire both pre- and postcourse. A one-way analysis of covariance controlling for gender, program (medical doctor versus physician assistant), and precourse responses demonstrated that enrollees had a significantly increased interest in primary care compared to nonenrollees after the course (F 1,32=9.22, P=.005). Enrollees also more positively rated their attitudes toward compensation, scope of practice, and job fulfillment than nonenrollees. Both groups had high levels of agreement on statements concerning patient-physician interactions and the importance of primary care to the health care system. CONCLUSION: The design and content of this elective offers a framework for other institutions looking to promote the value of primary care specialties, particularly family medicine. Creating opportunities for experiential learning and early student-faculty engagement may encourage preclerkship learners to consider a career in primary care.

Longitudinal trends in e-cigarette devices used by Californian youth, 2014-2018.

The rate of adolescent and young adult (AYA) e-cigarette usage has increased in recent years, possibly due to the introduction of sleek new e-cigarette devices such as JUUL. This study analyzed data from 400 California AYA to examine trends in e-cigarette usage by device type (disposables, large-size rechargeables, vape/hookah pens, JUUL/pod-based). Participants were asked about their ever, past 30-day, and past 7-day use of e-cigarettes; their usual e-cigarette device used; and co-use of devices in seven surveys administered approximately biannually from 2014 to 2018. During this time period, total e-cigarette ever-usage in our cohort increased linearly from 14.1% to 46.2% (ptrend < 0.001). JUUL/pod-based e-cigarette ever-usage increased from 14.9% to 22.5% in just six months in 2018. Furthermore, a majority of new e-cigarette users at the time of the survey endorsed using JUUL/pod-based devices (58.3% in Wave 6, 73.0% in Wave 7). With newer device options, AYA were also increasingly less likely to endorse older models such as disposables (19.1% to 6.9% from 2014 to 2018, ptrend < 0.01) and rechargeables (69.1% to 26.2% from 2014 to 2018, ptrend < 0.001) as their usual e-cigarette device. Participants who used JUUL/pod-based only as their usual device were more likely to endorse using only JUUL/pod-based devices during follow-up survey (70%), and none switched to a new device completely. Overall, this study provides a snapshot of how AYA's e-cigarette preferences appear to respond to new devices entering the market.

The Effect of Improving Cycleway Environment on the Recreational Benefits of Bicycle Tourism.

Bicycle tourism is one of the popular physical activities for sport tourists. Since the physical environment may affect bicycling behavior, it becomes an important determinant for cyclists to choose a cycleway. Exploratory factor analysis is performed to extract the perception of environmental quality of cyclists into five main factors, including safety, light facilities, lane design, landscape, and environment cleanliness. The contingent behavior method (CBM) is adopted to measure the quality improvement projects in different scenarios of light facility and landscape improvement. The results showed that the improvement projects increased the intended number of trips and the recreational benefits of cyclists.

Pre-diagnostic circulating insulin-like growth factor-I and bladder cancer risk in the European Prospective Investigation into Cancer and Nutrition.

Previous in vitro and case-control studies have found an association between the insulin-like growth factor (IGF)-axis and bladder cancer risk. Circulating concentrations of IGF-I have also been found to be associated with an increased risk of several cancer types; however, the relationship between pre-diagnostic circulating IGF-I concentrations and bladder cancer has never been studied prospectively. We investigated the association of pre-diagnostic plasma concentrations of IGF-I with risk of overall bladder cancer and urothelial cell carcinoma (UCC) in a case-control study nested within the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort. A total of 843 men and women diagnosed with bladder cancer between 1992 and 2005 were matched with 843 controls by recruitment centre, sex, age at recruitment, date of blood collection, duration of follow-up, time of day and fasting status at blood collection using an incidence density sampling protocol. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated using conditional logistic regression with adjustment for smoking status. No association was found between pre-diagnostic circulating IGF-I concentration and overall bladder cancer risk (adjusted OR for highest versus lowest fourth: 0.91, 95% CI: 0.66-1.24, ptrend = 0.40) or UCC (n of cases = 776; 0.91, 0.65-1.26, ptrend = 0.40). There was no significant evidence of heterogeneity in the association of IGF-I with bladder cancer risk by tumour aggressiveness, sex, smoking status, or by time between blood collection and diagnosis (pheterogeneity > 0.05 for all). This first prospective study indicates no evidence of an association between plasma IGF-I concentrations and bladder cancer risk.

Possible Facilitative Effects of Repeated Anodal Transcranial Direct Current Stimulation on Functional Outcome 1 Month Later in Schizophrenia: An Open Trial.

Recent research on neuromodulation techniques, such as transcranial direct current stimulation (tDCS), for the treatment of schizophrenia has mainly focused on psychotic symptoms. We aimed to determine whether repetitive tDCS is efficacious in improving determinants of outcome, such as cognitive function, daily living skills, and depressive mood in patients with schizophrenia. Twenty-eight patients underwent tDCS (2 mA × 20 min) two times per day for 5 consecutive days. The anodal electrode was placed over the left dorsolateral prefrontal cortex while the cathodal electrode was placed over the right supraorbital region. One month after the last stimulation, there was a significant improvement on cognitive function, measured by the brief assessment of cognition in schizophrenia (d = 0.49). Significant effects were also shown on daily living skills (functional capacity), measured by the UCSD performance-based skills assessment-brief (d = 0.70). Depressive symptoms, measured by the Calgary depression rating scale, as well as psychotic symptoms measured by on the positive and negative syndrome scale positive and general psychopathology subscales also responded to the treatment (d = 0.38, d = 0.48, and d = 0.50, respectively). This is the first study to suggest that tDCS with the anodal electrode on the left prefrontal cortex improves functional capacity and depressive symptoms in patients with schizophrenia. These results may add to the concept that tDCS provides a strategy to enhance functional outcomes in patients with schizophrenia. TRIAL REGISTRATION: https://upload.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000018556, UMIN000015953.

Macrophage Blockade Using CSF1R Inhibitors Reverses the Vascular Leakage Underlying Malignant Ascites in Late-Stage Epithelial Ovarian Cancer.

Malignant ascites is a common complication in the late stages of epithelial ovarian cancer (EOC) that greatly diminishes the quality of life of patients. Malignant ascites is a known consequence of vascular dysfunction, but current approved treatments are not effective in preventing fluid accumulation. In this study, we investigated an alternative strategy of targeting macrophage functions to reverse the vascular pathology of malignant ascites using fluid from human patients and an immunocompetent murine model (ID8) of EOC that mirrors human disease by developing progressive vascular disorganization and leakiness culminating in massive ascites. We demonstrate that the macrophage content in ascites fluid from human patients and the ID8 model directly correlates with vascular permeability. To further substantiate macrophages' role in the pathogenesis of malignant ascites, we blocked macrophage function in ID8 mice using a colony-stimulating factor 1 receptor kinase inhibitor (GW2580). Administration of GW2580 in the late stages of disease resulted in reduced infiltration of protumorigenic (M2) macrophages and dramatically decreased ascites volume. Moreover, the disorganized peritoneal vasculature became normalized and sera from GW2580-treated ascites protected against endothelial permeability. Therefore, our findings suggest that macrophage-targeted treatment may be a promising strategy toward a safe and effective means to control malignant ascites of EOC.

Interleukin-32 increases human gastric cancer cell invasion associated with tumor progression and metastasis.

PURPOSE: The proinflammatory cytokine interleukin-32 (IL-32) is a novel tumor marker highly expressed in various human carcinomas, including gastric cancer. However, its effects on prognosis of patients with gastric cancer and cancer metastasis are virtually unknown at present. The main aim of this study was to explore the clinical significance of IL-32 in gastric cancer and further elucidate the molecular mechanisms underlying IL-32-mediated migration and invasion. EXPERIMENTAL DESIGN: Gastric cancer cells with ectopic expression or silencing of IL-32 were examined to identify downstream molecules and establish their effects on cell motility, invasion, and lung metastasis in vivo. RESULTS: IL-32 was significantly upregulated in gastric cancer and positively correlated with aggressiveness of cancer and poor prognosis. Ectopic expression of IL-32 induced elongated morphology and increased cell migration and invasion via induction of IL-8, VEGF, matrix metalloproteinase 2 (MMP2), and MMP9 expression via phosphor-AKT/phospho-glycogen synthase kinase 3ß/active ß-catenin as well as hypoxia-inducible factor 1α (HIF-1α) signaling pathways. Conversely, depletion of IL-32 in gastric cancer cells reversed these effects and decreased lung colonization in vivo. Examination of gene expression datasets in oncomine and staining of gastric cancer specimens demonstrated the clinical significance of IL-32 and its downstream molecules by providing information on their coexpression patterns. CONCLUSIONS: IL-32 contributes to gastric cancer progression by increasing the metastatic potential resulting from AKT, ß-catenin, and HIF-1α activation. Our results clearly suggest that IL-32 is an important mediator for gastric cancer metastasis and independent prognostic predictor of gastric cancer.

Thyroid hormone regulation of miR-21 enhances migration and invasion of hepatoma.

Thyroid hormone (T(3)) signaling through the thyroid hormone receptor (TRα1) regulates hepatoma cell growth and pathophysiology, but the underlying mechanisms are unclear at present. Here, we have shown that the oncomir microRNA-21 (miR-21) is activated by T(3) through a native T(3) response element in the primary miR-21 promoter. Overexpression of miR-21 promoted hepatoma cell migration and invasion, similar to that observed with T(3) stimulation in hepatoma cells. In addition, anti-miR-21-induced suppression of cell migration was rescued by T(3). The Rac-controlled regulator of invasion and metastasis, T-cell lymphoma invasion and metastasis 1 (TIAM1), was identified as a miR-21 target additionally downregulated by T(3). Attenuation and overexpression of miR-21 induced upregulation and downregulation of TIAM1, respectively. TIAM1 attenuation, in turn, enhanced migration and invasion via the upregulation of ß-catenin, vimentin, and matrix metalloproteinase-2 in hepatoma cells. Notably, correlations between TRα1, miR-21, and TIAM1 expression patterns in animal models paralleled those observed in vitro. In the clinic, we observed a positive correlation (P = 0.005) between the tumor/nontumor ratios of TRα1 and miR-21 expression, whereas a negative correlation (P = 0.019) was seen between miR-21 and TIAM1 expression in patients with hepatoma. Our findings collectively indicate that miR-21 stimulation by T(3) and subsequent TIAM1 suppression promotes hepatoma cell migration and invasion.

Thyroid hormone actions in liver cancer.

The thyroid hormone 3,3',5-triiodo-L-thyronine (T3) mediates several physiological processes, including embryonic development, cellular differentiation, metabolism, and the regulation of cell proliferation. Thyroid hormone receptors (TRs) generally act as heterodimers with the retinoid X receptor (RXR) to regulate target genes. In addition to their developmental and metabolic functions, TRs have been shown to play a tumor suppressor role, suggesting that their aberrant expression can lead to tumor transformation. Conversely, recent reports have shown an association between overexpression of wild-type TRs and tumor metastasis. Signaling crosstalk between T3/TR and other pathways or specific TR coregulators appear to affect tumor development. Since TR actions are complex as well as cell context-, tissue- and time-specific, aberrant expression of the various TR isoforms has different effects during diverse tumorigenesis. Therefore, elucidation of the T3/TR signaling mechanisms in cancers should facilitate the identification of novel therapeutic targets. This review provides a summary of recent studies focusing on the role of TRs in hepatocellular carcinomas (HCCs).

Criterion and construct validity of the CogState Schizophrenia Battery in Japanese patients with schizophrenia.

BACKGROUND: The CogState Schizophrenia Battery (CSB), a computerized cognitive battery, covers all the same cognitive domains as the Measurement and Treatment Research to Improve Cognition in Schizophrenia (MATRICS) Consensus Cognitive Battery but is briefer to conduct. The aim of the present study was to evaluate the criterion and construct validity of the Japanese language version of the CSB (CSB-J) in Japanese patients with schizophrenia. METHODOLOGY/PRINCIPAL FINDINGS: Forty Japanese patients with schizophrenia and 40 Japanese healthy controls with matching age, gender, and premorbid intelligence quotient were enrolled. The CSB-J and the Brief Assessment of Cognition in Schizophrenia, Japanese-language version (BACS-J) were performed once. The structure of the CSB-J was also evaluated by a factor analysis. Similar to the BACS-J, the CSB-J was sensitive to cognitive impairment in Japanese patients with schizophrenia. Furthermore, there was a significant positive correlation between the CSB-J composite score and the BACS-J composite score. A factor analysis showed a three-factor model consisting of memory, speed, and social cognition factors. CONCLUSIONS/SIGNIFICANCE: This study suggests that the CSB-J is a useful and rapid automatically administered computerized battery for assessing broad cognitive domains in Japanese patients with schizophrenia.