Weakened APC/C activity at mitotic exit drives cancer vulnerability to KIF18A inhibition.

The efficacy of current antimitotic cancer drugs is limited by toxicity in highly proliferative healthy tissues. A cancer-specific dependency on the microtubule motor protein KIF18A therefore makes it an attractive therapeutic target. Not all cancers require KIF18A, however, and the determinants underlying this distinction remain unclear. Here, we show that KIF18A inhibition drives a modest and widespread increase in spindle assembly checkpoint (SAC) signaling from kinetochores which can result in lethal mitotic delays. Whether cells arrest in mitosis depends on the robustness of the metaphase-to-anaphase transition, and cells predisposed with weak basal anaphase-promoting complex/cyclosome (APC/C) activity and/or persistent SAC signaling through metaphase are uniquely sensitive to KIF18A inhibition. KIF18A-dependent cancer cells exhibit hallmarks of this SAC:APC/C imbalance, including a long metaphase-to-anaphase transition, and slow mitosis overall. Together, our data reveal vulnerabilities in the cell division apparatus of cancer cells that can be exploited for therapeutic benefit.

Quantifying full-length circular RNAs in cancer.

Circular RNAs (circRNAs) are abundantly expressed in cancer. Their resistance to exonucleases enables them to have potentially stable interactions with different types of biomolecules. Alternative splicing can create different circRNA isoforms that have different sequences and unequal interaction potentials. The study of circRNA function thus requires knowledge of complete circRNA sequences. Here we describe psirc, a method that can identify full-length circRNA isoforms and quantify their expression levels from RNA sequencing data. We confirm the effectiveness and computational efficiency of psirc using both simulated and actual experimental data. Applying psirc on transcriptome profiles from nasopharyngeal carcinoma and normal nasopharynx samples, we discover and validate circRNA isoforms differentially expressed between the two groups. Compared with the assumed circular isoforms derived from linear transcript annotations, some of the alternatively spliced circular isoforms have 100 times higher expression and contain substantially fewer microRNA response elements, showing the importance of quantifying full-length circRNA isoforms.

The changes in biethnic acceptance across daily settings and biethnic adolescents' psychological well-being during middle school transition.

INTRODUCTION: Patterns of biethnic adolescents' perceived biethnic acceptance across families, peers, and school contexts were examined during the transition from elementary to middle school in South Korea. We also examined how the transition patterns were related to their psychological outcomes during this period. METHODS: Utilizing 2-wave data (2017 and 2019) from the Panel Survey of Korean Multicultural Youth Adjustment, a latent transition analysis was conducted. Participants were biethnic adolescents who were in 5th or 6th grade at Wave 1 (N = 245; 51.02% female; Mage = 11.38). Their fathers were Korean, and mothers were immigrants from neighboring countries. Familial ethnic socialization, peer discrimination, and school multicultural climate scores were used as indicators of biethnic acceptance. Outcomes of self-esteem, depression, and biethnic affirmation were also examined. RESULTS: Latent profile and transition analyses yielded two groups (i.e., high acceptance and low acceptance) at each wave and four transition patterns (i.e., high-high, low-high, low-low, and high-low). Compared to high-high group, which was the most prevalent group, low-low and high-low groups reported lower self-esteem and ethnic affirmation, and greater depression at Wave 3. CONCLUSIONS: While for the majority of participants, their daily settings continued to be high in biethnic acceptance across the transition period, most at risk were those who perceived a decrease in biethnic acceptance in their daily settings. Results shed light on the need for support to maintain the context of high biethnic acceptance surrounding biethnic adolescents for their psychological well-being in school transitions.

Role of nutrient-driven O-GlcNAc-post-translational modification in pancreatic exocrine and endocrine islet development.

Although the developing pancreas is exquisitely sensitive to nutrient supply in utero, it is not entirely clear how nutrient-driven post-translational modification of proteins impacts the pancreas during development. We hypothesized that the nutrient-sensing enzyme O-GlcNAc transferase (Ogt), which catalyzes an O-GlcNAc-modification onto key target proteins, integrates nutrient-signaling networks to regulate cell survival and development. In this study, we investigated the heretofore unknown role of Ogt in exocrine and endocrine islet development. By genetic manipulation in vivo and by using morphometric and molecular analyses, such as immunofluorescence imaging and single cell RNA sequencing, we show the first evidence that Ogt regulates pancreas development. Genetic deletion of Ogt in the pancreatic epithelium (OgtKOPanc) causes pancreatic hypoplasia, in part by increased apoptosis and reduced levels of of Pdx1 protein. Transcriptomic analysis of single cell and bulk RNA sequencing uncovered cell-type heterogeneity and predicted upstream regulator proteins that mediate cell survival, including Pdx1, Ptf1a and p53, which are putative Ogt targets. In conclusion, these findings underscore the requirement of O-GlcNAcylation during pancreas development and show that Ogt is essential for pancreatic progenitor survival, providing a novel mechanistic link between nutrients and pancreas development.

Dietary intakes of branched-chain amino acids and plasma lipid profiles among filipino women in Korea: the Filipino Women's Diet and Health Study (FiLWHEL).

BACKGROUND: The potential role of dietary branched-chain amino acids (BCAA) in metabolic health, including cardiovascular disease and diabetes, is evolving, and it is yet to be understood if dietary BCAA intakes are associated with plasma lipid profiles or dyslipidaemia. This study tested the association of dietary BCAA intakes with plasma lipid profiles and dyslipidaemia among Filipino women in Korea. METHODS: Energy-adjusted dietary BCAA intakes (isoleucine, leucine, valine, and total BCAA) and fasting blood profiles of triglycerides (TG), total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), and low-density lipoprotein-cholesterol (LDL-C) were determined in a sample of 423 women enrolled in the Filipino Women's Diet and Health Study (FiLWHEL). The generalized linear model was applied to estimate least-square (LS) means and 95% confidence intervals (CIs) and compare plasma TG, TC, HDL-C, and LDL-C across tertile distribution of energy-adjusted dietary BCAA intakes at P < 0.05. RESULTS: Mean of energy-adjusted dietary total BCAA intake was 8.3 ± 3.9 g/d. Average plasma lipid profiles were 88.5 ± 47.4 mg/dl for TG, 179.7 ± 34.5 mg/dl for TC, 58.0 ± 13.7 mg/dl for HDL-C, and 104.0 ± 30.5 mg/dl for LDL-C. LS means, and 95% CIs across tertiles of energy-adjusted total BCAA intakes were 89.9 mg/dl, 88.8 mg/dl and 85.8 mg/dl (P-trend = 0.45) for TG, 179.1 mg/dl, 183.6 mg/dl and 176.5 mg/dl (P-trend = 0.48) for TC, 57.5 mg/dl, 59.6 mg/dl and 57.1 mg/dl (P-trend = 0.75) for HDL-C and 103.6 mg/dl, 106.2 mg/dl and 102.3 mg/dl (P-trend = 0.68) for LDL-C. Furthermore, the multivariable-adjusted prevalence ratios and 95% confidence intervals for dyslipidaemia across increasing tertile distribution of energy-adjusted total BCAA intake were; 1.00, 0.67 (0.40, 1.13) and 0.45 (0.16, 1.27; P-trend = 0.03) for the first, second and third tertile, respectively. CONCLUSIONS: Higher dietary intakes of BCAA presented a statistically significant inverse trend with the prevalence of dyslipidaemia among Filipino women in this study and testing these associations in longitudinal studies may be necessary to confirm these findings.

Comparison of cardiovascular disease risk factors among FiLWHEL (2014-2016), NNS (2013) and KNHANES (2013-2015) women.

OBJECTIVES: This study assessed the CVD risk factors among Filipino women (FW) in Korea and compared them with FW in the Philippines and women in Korea (KW). METHODS: A cohort of 504 women from the Filipino Women's Health and Diet Study (FiLWHEL) aged 20-57 years old were age-matched (1:1 ratio) with women from the 2013 National Nutrition Survey in the Philippines and the 2013-2015 Korean National Health and Nutrition Examination Survey. Anthropometric data, blood pressure (BP), lipid and glucose levels were compared across the four populations by calculating the odds ratio (OR)s and 95% confidence interval (CI)s using conditional logistic regression models. RESULTS: Compared to KW, FW in Korea and FW in the Philippines were more than 2 and 3 times higher odds of having obesity for BMI ≥ 30 kg/m2 and waist circumference ≥ 88 cm, respectively. However, FW in Korea had the highest odds (OR 5.51, 95% CI 3.18-9.56) of having hypertension compared to KW. FW in the Philippines had the highest odds of having dyslipidemia (compared to KW, total cholesterol ≥ 200 mg/dL: OR 8.83, 95% CI 5.30-14.71; LDL-C ≥ 130 mg/dL: OR 3.25, 95% CI 2.13-4.98; and triglyceride ≥ 150 mg/dL: OR 2.59, 95% CI 1.59-4.22), but FW in Korea and KW had similar prevalence of dyslipidemia. CONCLUSIONS: FW in Korea had higher prevalence of obesity and hypertension, with similar prevalence of dyslipidemia compared to KW in this sample. FW in the Philippines had higher prevalence of dyslipidemia compared to FW in Korea. Further prospective studies are warranted to examine the CVD risk factors among continental and native-born Filipino women.

Aspirin for Secondary Prevention of Cardiovascular Disease in 51 Low-, Middle-, and High-Income Countries.

Importance: Aspirin is an effective and low-cost option for reducing atherosclerotic cardiovascular disease (CVD) events and improving mortality rates among individuals with established CVD. To guide efforts to mitigate the global CVD burden, there is a need to understand current levels of aspirin use for secondary prevention of CVD. Objective: To report and evaluate aspirin use for secondary prevention of CVD across low-, middle-, and high-income countries. Design, Setting, and Participants: Cross-sectional analysis using pooled, individual participant data from nationally representative health surveys conducted between 2013 and 2020 in 51 low-, middle-, and high-income countries. Included surveys contained data on self-reported history of CVD and aspirin use. The sample of participants included nonpregnant adults aged 40 to 69 years. Exposures: Countries' per capita income levels and world region; individuals' socioeconomic demographics. Main Outcomes and Measures: Self-reported use of aspirin for secondary prevention of CVD. Results: The overall pooled sample included 124 505 individuals. The median age was 52 (IQR, 45-59) years, and 50.5% (95% CI, 49.9%-51.1%) were women. A total of 10 589 individuals had a self-reported history of CVD (8.1% [95% CI, 7.6%-8.6%]). Among individuals with a history of CVD, aspirin use for secondary prevention in the overall pooled sample was 40.3% (95% CI, 37.6%-43.0%). By income group, estimates were 16.6% (95% CI, 12.4%-21.9%) in low-income countries, 24.5% (95% CI, 20.8%-28.6%) in lower-middle-income countries, 51.1% (95% CI, 48.2%-54.0%) in upper-middle-income countries, and 65.0% (95% CI, 59.1%-70.4%) in high-income countries. Conclusion and Relevance: Worldwide, aspirin is underused in secondary prevention, particularly in low-income countries. National health policies and health systems must develop, implement, and evaluate strategies to promote aspirin therapy.

Interactions between the apolipoprotein E4 gene and modifiable risk factors for cognitive impairment: a nationally representative panel study.

BACKGROUND: Few studies using rigorous clinical diagnosis have considered whether associations with cognitive decline are potentiated by interactions between genetic and modifiable risk factors. Given the increasing burden of cognitive impairment (CI) and dementia, we assessed whether Apolipoprotein E ε4 (APOE4) genotype status modifies the association between incident CI and key modifiable risk factors . METHODS: Older adults (70+) in the US were included. APOE4 status was genotyped. Risk factors for CI were self-reported. Cognitive status (normal, CI, or dementia) was assigned by clinical consensus panel. In eight separate Cox proportional hazard models, we assessed for interactions between APOE4 status and other CI risk factors. RESULT: The analytical sample included 181 participants (mean age 77.7 years; 45.9% male). APOE4 was independently associated with a greater hazard of CI in each model (Hazard Ratios [HR] between 1.81-2.66, p < 0.05) except the model evaluating educational attainment (HR 1.65, p = 0.40). The joint effects of APOE4 and high school education or less (HR 2.25, 95% CI: 1.40-3.60, p < 0.001), hypertension (HR 2.46, 95% CI: 1.28-4.73, p = 0.007), elevated depressive symptoms (HR 5.09, 95% CI: 2.59-10.02, p < 0.001), hearing loss (HR 3.44, 95% CI: 1.87-6.33, p < 0.0001), vision impairment (HR 5.14, 95% CI: 2.31-11.43, p < 0.001), smoking (HR 2.35, 95% CI: 1.24-4.47, p = 0.009), or obesity (HR 3.80, 95% CI: 2.11-6.85, p < 0.001) were associated with the hazard of incident CIND (compared to no genetic or modifiable risk factor) in separate models. The joint effect of Apolipoprotein ε4 and type 2 diabetes was not associated with CIND (HR 1.58, 95% CI: 0.67-2.48, p = 0.44). DISCUSSION: The combination of APOE4 and selected modifiable risk factors conveys a stronger association with incident CI than either type of risk factor alone.

Dietary branched-chain amino acids and odds of obesity among immigrant Filipino women: the Filipino women's diet and health study (FiLWHEL).

BACKGROUND: The dietary environment promoting adiposity keeps evolving and of interest is the significance of dietary branched-chain amino acids (BCAA). This study assessed the association between dietary BCAA intakes and odds of obesity among immigrant Filipino women in Korea. METHOD: We included 423 immigrant Filipino women enrolled in the Filipino Women's diet and health study in the Republic of Korea. Dietary BCAA intakes were estimated from 24 hour recalls and adjusted for energy intake using the residual method. General obesity was derived from direct anthropometric measurements (height, weight and waist circumference - WC) and defined as body mass index (BMI) ≥25 kg/m2 and abdominal obesity as WC ≥80 cm. Odds ratios (OR) and 95% confidence intervals (CI) by tertile distribution of energy-adjusted BCAA intakes were estimated using multivariable logistic regression with a two-sided P < 0.05. RESULTS: Median (interquartile range) for BCAA intakes in g/day were; 7.9 (5.8, 10.3) g/day for total BCAA; 2.0 (1.5, 2.6) g/day for isoleucine, 3.5(2.5, 4.6) g/day for leucine and 2.4 (1.8, 3.1) g/day for valine. Mean BMI and WC were 23.6 ± 3.8 kg/m2 and 79.8 ± 9.3 cm, respectively. Also, 30.2% (128) had BMI ≥25 kg/m2 and 42.0% (178) had WC ≥80 cm. ORs (95%CIs) of general obesity across tertile distribution of energy-adjusted total BCAA intakes were 1.00, 0.81 (0.47, 1.37) and 0.62 (0.36, 1.07; P for trend = 0.08). A similar trend was observed across tertile distribution of energy-adjusted isoleucine, leucine and valine intakes. However, there was a statistically significant inverse association between total BCAA intake and odds of general obesity in a subset of non-smokers; 1.00, 0.68 (0.39, 1.20) and 0.55 (0.31, 0.98; P for trend = 0.04). CONCLUSION: We found a suggestive inverse association between higher dietary BCAA intake and odds of obesity in this sample of immigrant Filipino women, particularly among non-smokers. Prospective cohort studies among the immigrant population will be necessary to verity these findings.

Effect of preemptive topical application of lidocaine or prilocaine-lidocaine on successful catheterization of ear veins in New Zealand White rabbits.

OBJECTIVE: To compare 5% lidocaine ointment applied for 10 minutes before intravenous (IV) catheterization to a eutectic mixture of local anesthetics (2.5% lidocaine and 2.5% prilocaine; EMLA) cream applied for 10 or 60 minutes prior and a water-based placebo for 60 minutes for successful marginal ear IV catheterization in rabbits. STUDY DESIGN: Randomized, blinded, complete crossover, controlled trial. ANIMALS: A total of 10 intact male, New Zealand White rabbits aged 4 months. METHODS: After application of one of the treatments on the marginal ear vein, an operator unaware of the treatment attempted IV catheter placement. Successful IV catheterization was considered as a catheter inserted at the first attempt, safely secured, with appropriate patency. Generalized linear mixed models were developed to identify relevant predictors of successful catheter placement. RESULTS: Ears that had EMLA cream applied 60 minutes before the catheterization attempt had more than 10 times the odds of successful catheter placement [odds ratio (OR)=10.75; 95% confidence interval (CI), 1.92-60.16; p < 0.008] compared with placebo. Both the application of EMLA cream or lidocaine ointment 10 minutes before the catheterization attempt resulted in approximately three times the odds of successful catheter placement compared with placebo, but were not statistically significant. CONCLUSIONS AND CLINICAL RELEVANCE: Application of EMLA cream on the marginal ear vein 60 minutes before catheterization increases the chance of successful IV catheter placement in conscious rabbits.

Student Experience and Ethnic Diversity: The Experiences of Underrepresented Minority Students at a Veterinary University in the United Kingdom.

Despite a lack of diversity, studies on the experience of ethnically diverse (ED) students have not focused on veterinary education. This study used focus groups to explore the experience of ED veterinary students, their challenges, and their sources of support in this setting. Focus groups were held using two formats: a traditional, in-person approach, and online, via a social media group. Recruitment was by invitation and focus group allocation according to participant preference. Conversations were transcribed or downloaded, anonymized, and analyzed using a two-part narrative analysis, the aim of which was to understand the experiences of ED veterinary students at an in-depth level. Students experienced identity conflicts at multiple levels (self, others, profession), which appeared to relate to their ethnicity. Conflict between self and others resulted in a feeling of otherness arising from a White student majority, both within the university and on external placements, and professional identity conflicts arose between students' personal cultural values and their values as a veterinarian. Internal conflicts arose when students felt a wish to integrate but also perceived a need to segregate with similar others to obtain support and a sense of belonging to a group. These challenges have potential implications for mental well-being and career opportunities. For veterinary medicine to adapt to changing client demographics in an increasingly globalized world, a deeper understanding of the ED student experience may offer advantages in areas such as recruitment and profession retention, which will eventually support greater diversity within the professional population.

Pharmacy stakeholder reports on ethical and logistical considerations in anti-opioid vaccine development.

BACKGROUND: As opioid use disorder (OUD) incidence and its associated deaths continue to persist at elevated rates, the development of novel treatment modalities is warranted. Recent strides in this therapeutic area include novel anti-opioid vaccine approaches. This work compares logistical and ethical considerations surrounding currently available interventions for opioid use disorder with an anti-opioid vaccine approach. METHODS: The opinions of student pharmacists and practicing pharmacists assessing knowledge, perceptions, and attitudes toward current and future OUD management strategies were characterized using a staged, multi-modal research approach incorporating a focus group, pilot survey development and refinement, and final survey deployment. Survey responses were assessed using one- and two-way parametric and non-parametric analyses where appropriate, and multi-dimensional matrix profiles were compared using z-tests following an exhaustive combinatorial sum of differences calculation between items within each compared matrix. RESULTS: Focus group content analysis revealed a high level of agreeableness among participants regarding anti-opioid vaccine technology and a sense of shared ownership regarding solutions to the opioid epidemic at large. Pilot survey results demonstrated subject ability to consider both pragmatic and ethical considerations related to current therapeutics and novel interventions in a single instrument, with high endurance amongst engaged subjects. Access inequality was the most concerning ethical consideration identified for anti-opioid vaccines. Support for anti-opioid vaccine implementation across various clinical scenarios was strongest for voluntary use amongst individuals in recovery, and lowest for mandatory use in at-risk individuals. CONCLUSIONS: Ethical and logistical concerns surrounding anti-opioid vaccines were largely similar to those for current OUD therapeutics overall. Anti-opioid vaccines were endorsed as helpful potential additions to current OUD therapeutic approaches, particularly for voluntary use in the later stages of clinical progression.

Adolescents' Daily Experience of Fathers' Emotional Support and Self-Evaluation in Korea.

Recently, fathers' role in children's development has been recognized internationally. In Asian countries, similar conversations have emerged but there has been a lack of empirical studies that considered the unique cultural contexts. As a response, based on Bronfenbrenner's process-person-context-time model, the present study examined the daily dynamics and individual differences in the experience of fathers' emotional support and self-evaluation in social roles among 283 Korean adolescents in 5th and 8th grade. Through Hierarchical Linear Modeling, within-person associations between adolescents' experience of fathers' emotional support and self-evaluation in social roles on the same day and the next day were found, with individual differences by grade level and family affluence. Theoretical and practical implications within the relevant cultural context are discussed.

Context of biethnic acceptance, biethnic affirmation, and life satisfaction among Korean biethnic adolescents.

INTRODUCTION: This study aimed to identify latent profiles based on familial ethnic socialization, peer discrimination, and school multicultural climate among biethnic adolescents in South Korea and to examine how the emerged profiles were related to their life satisfaction.We also examined how biethnic affirmation interacted with the profiles in predicting life satisfaction. METHODS: Questionnaire data from the first wave of the Panel Survey of Korean Multicultural Youth Adjustment was used. The sample consisted of 358 biethnic adolescents in 5th and 6th grade in South Korea. The meanage was 11.41 years old. Approximately 60% of the participants were in 5th grade and 51.68% were girls. RESULTS: The results of LPA showed that familial ethnic socialization, peer discrimination, and school multicultural climate formed two distinct profiles in the current sample. These profiles were characterized as contexts of high and low biethnic acceptance. Biethnic adolescents in the context of high biethnic acceptance were significantly more satisfied with their lives than those in the context of low biethnic acceptance.Further, the positive association between biethnic iaffirmation and life satisfaction was more pronounced among adolescents in the context of low biethnic acceptance. CONCLUSIONS: Findings suggest the importance of promoting biethnic acceptance across family, peer, and school settings for adolescents' biethnic affirmation and life satisfaction.

Gestational Diabetes Mellitus: A Harbinger of the Vicious Cycle of Diabetes.

Gestational diabetes mellitus (GDM), characterized by a transitory form of diabetes induced by insulin resistance and pancreatic ß-cell dysfunction during pregnancy, has been identified as one of the major obstacles in achieving improved maternal and child health. Approximately 9-25% of pregnancies worldwide are impacted by the acute, long-term, and transgenerational health complications of this disease. Here, we discuss how GDM affects longstanding maternal and neonatal outcomes, as well as health risks that likely persist into future generations. In addition to the current challenges in the management and diagnosis of and the complications associated with GDM, we discuss current preclinical models of GDM to better understand the underlying pathophysiology of the disease and the timely need to increase our scientific toolbox to identify strategies to prevent and treat GDM, thereby advancing clinical care.

Oncogenic S1P signalling in EBV-associated nasopharyngeal carcinoma activates AKT and promotes cell migration through S1P receptor 3.

Undifferentiated nasopharyngeal carcinoma (NPC) is a cancer with high metastatic potential that is consistently associated with Epstein-Barr virus (EBV) infection. In this study, we have investigated the functional contribution of sphingosine-1-phosphate (S1P) signalling to the pathogenesis of NPC. We show that EBV infection or ectopic expression of the EBV-encoded latent genes (EBNA1, LMP1, and LMP2A) can up-regulate sphingosine kinase 1 (SPHK1), the key enzyme that produces S1P, in NPC cell lines. Exogenous addition of S1P promotes the migration of NPC cells through the activation of AKT; shRNA knockdown of SPHK1 resulted in a reduction in the levels of activated AKT and inhibition of cell migration. We also show that S1P receptor 3 (S1PR3) mRNA is overexpressed in EBV-positive NPC patient-derived xenografts and a subset of primary NPC tissues, and that knockdown of S1PR3 suppressed the activation of AKT and the S1P-induced migration of NPC cells. Taken together, our data point to a central role for EBV in mediating the oncogenic effects of S1P in NPC and identify S1P signalling as a potential therapeutic target in this disease. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

MicroRNA-183 suppresses cancer stem-like cell properties in EBV-associated nasopharyngeal carcinoma.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV)-associated epithelial malignancy that exhibits distinct geographical and ethnic prevalence. Although the contemporary therapeutic approach of radio-/chemotherapy provides excellent results for patients with early-stage disease, it is far from satisfactory for those with disease remission and distant metastasis. Promising therapeutic strategies for advanced and relapsed NPC are still lacking. We recently identified and characterized a cancer stem-like cell (CSC) subpopulation in NPC that appeared to play an important role in tumor progression. Microarray analysis revealed downregulation of several stemness-inhibiting miRNAs in these CSC cells. Among these miRNAs, miR-96 and miR-183 showed the highest fold change and were selected to elucidate their role in repressing NPC CSC properties. METHODS: MiR-96 and miR-183 expression in NPC CSCs was detected by qRT-PCR. Transient and stable transfection was performed in EBV-positive NPC C666-1 cells to examine the effects of ectopic expression of miR-96 and miR-183 on repressing cell growth and CSC properties. Anchorage-dependent (colony formation) and anchorage-independent (tumor sphere formation) growths of these miR-96 and miR-183 expressing cells were determined. Expression of multiple CSC markers and related molecules were accessed by flow cytometry and Western blotting. The tumorigenicity of the stable miR-96- and miR-183-transfected NPC cells was examined in an in vivo nude mice model. RESULTS: Downregulation of miR-96 and miR-183 was confirmed in NPC spheroids. Using transient or stable transfection, we showed that ectopic expression of miR-96 and miR-183 suppressed cell growth and tumor sphere formation in NPC. Reduced NICD3 and NICD4 in miR-96- and miR-183-expressing NPC cells suggests the involvement of the NOTCH signaling pathway in their tumor suppressive function. Finally, we showed that the tumorigenicity of cells stably expressing miR-183 was significantly inhibited in the in vivo nude mice model. CONCLUSIONS: miR-183 is a tumor-suppressive miRNA in EBV-associated NPC. Its abilities to suppress CSC properties in vitro and effectively reduce tumor growth in vivo shed light on its role as a potential therapeutic target.

US Physicians' Opinions about Distinctions between Withdrawing and Withholding Life-Sustaining Treatment.

Decisions to withhold or withdraw life-sustaining treatment (LST) precede the majority of ICU deaths. Although professional guidelines generally treat the two as ethically equivalent, evidence suggests withdrawing LST is often more psychologically difficult than withholding it. The aim of the experiment was to investigate whether physicians are more supportive of withholding LST than withdrawing it and to assess how physicians' opinions are shaped by their religious characteristics, specialty, and experience caring for dying patients. In 2010, a survey was mailed to 2016 practicing US physicians. Physicians were asked whether physicians should always comply with a competent patient's request to withdraw LST, whether withdrawing LST is more psychologically difficult than withholding it, and whether withdrawing LST is typically more ethically problematic than withholding it. Of 1880 eligible physicians, 1156 responded to the survey (62%); 93% agreed that physicians should always comply with a competent patient's request to withdraw LST. More than half of the physicians reported that they find withdrawing LST more psychologically difficult than withholding it (61%), and that withdrawing LST is typically more ethically problematic (59%). Physician religiosity was associated with finding withdrawal more ethically problematic, but not with finding it more psychologically difficult. Physicians working in an end-of-life specialty and physicians with more experience caring for dying patients were less likely to endorse either a psychological or an ethical distinction between withdrawing and withholding LST. Most US physicians find withdrawing LST not only more psychologically difficult, but also more ethically problematic than withholding such treatment. Physicians' opinions are to some extent shaped by their religious characteristics, specialty, and levels of experience caring for dying patients.

Copy number gain of granulin-epithelin precursor (GEP) at chromosome 17q21 associates with overexpression in human liver cancer.

BACKGROUND: Granulin-epithelin precursor (GEP), a secretory growth factor, demonstrated overexpression in various human cancers, however, mechanism remain elusive. Primary liver cancer, hepatocellular carcinoma (HCC), ranks the second in cancer-related death globally. GEP controlled growth, invasion, metastasis and chemo-resistance in liver cancer. Noted that GEP gene locates at 17q21 and the region has been frequently reported to be amplified in subset of HCC. The study aims to investigate if copy number gain would associate with GEP overexpression. METHODS: Quantitative Microsatellite Analysis (QuMA) was used to quantify the GEP DNA copy number, and fluorescent in situ hybridization (FISH) was performed to consolidate the amplification status. GEP gene copy number, mRNA expression level and clinico-pathological features were analyzed. RESULTS: GEP DNA copy number determined by QuMA corroborated well with the FISH data, and the gene copy number correlated with the expression levels (n = 60, r = 0.331, P = 0.010). Gain of GEP copy number was observed in 20% (12/60) HCC and associated with hepatitis B virus infection status (P = 0.015). In HCC with increased GEP copy number, tight association between GEP DNA and mRNA levels were observed (n = 12, r = 0.664, P = 0.019). CONCLUSIONS: Gain of the GEP gene copy number was observed in 20% HCC and the frequency comparable to literatures reported on the chromosome region 17q. Increased gene copy number contributed to GEP overexpression in subset of HCC.