skNAC, a Smyd1-interacting transcription factor, is involved in cardiac development and skeletal muscle growth and regeneration.

Cardiac and skeletal muscle development and maintenance require complex interactions between DNA-binding proteins and chromatin remodeling factors. We previously reported that Smyd1, a muscle-restricted histone methyltransferase, is essential for cardiogenesis and functions with a network of cardiac regulatory proteins. Here we show that the muscle-specific transcription factor skNAC is the major binding partner for Smyd1 in the developing heart. Targeted deletion of skNAC in mice resulted in partial embryonic lethality by embryonic day 12.5, with ventricular hypoplasia and decreased cardiomyocyte proliferation that were similar but less severe than in Smyd1 mutants. Expression of Irx4, a ventricle-specific transcription factor down-regulated in hearts lacking Smyd1, also depended on the presence of skNAC. Viable skNAC(-/-) adult mice had reduced postnatal skeletal muscle growth and impaired regenerative capacity after cardiotoxin-induced injury. Satellite cells isolated from skNAC(-/-) mice had impaired survival compared with wild-type littermate satellite cells. Our results indicate that skNAC plays a critical role in ventricular cardiomyocyte expansion and regulates postnatal skeletal muscle growth and regeneration in mice.

Role of healthcare in Korean long-term care insurance.

With the rapid aging of the population, Korea introduced public long-term care insurance for older people in 2008. The long-term care insurance was designed as a separate scheme from the national health insurance, with eligibility qualifications and the certification process based on functional disability, benefits and coverage of community-based and institutional care, and a financing structure through multi-party contributions. Delivering appropriate health services to long-term care beneficiaries who manifest a high prevalence of comorbid chronic conditions with rising healthcare costs, however, presents a particular challenge. The lack of coordination between the health and long-term care sectors, limited consideration of physicians' assessments in the certification process, inadequate provision of health services in long-term care facilities, and overlapping and inefficient use of care resources act as barriers to providing comprehensive healthcare for older beneficiaries. Through active participation in the long-term care system, health professionals can help older patients navigate through the complex long-term care terrain to obtain quality healthcare.

Identification of a transporter complex responsible for the cytosolic entry of nitrogen-containing bisphosphonates.

Nitrogen-containing-bisphosphonates (N-BPs) are a class of drugs widely prescribed to treat osteoporosis and other bone-related diseases. Although previous studies have established that N-BPs function by inhibiting the mevalonate pathway in osteoclasts, the mechanism by which N-BPs enter the cytosol from the extracellular space to reach their molecular target is not understood. Here, we implemented a CRISPRi-mediated genome-wide screen and identified SLC37A3 (solute carrier family 37 member A3) as a gene required for the action of N-BPs in mammalian cells. We observed that SLC37A3 forms a complex with ATRAID (all-trans retinoic acid-induced differentiation factor), a previously identified genetic target of N-BPs. SLC37A3 and ATRAID localize to lysosomes and are required for releasing N-BP molecules that have trafficked to lysosomes through fluid-phase endocytosis into the cytosol. Our results elucidate the route by which N-BPs are delivered to their molecular target, addressing a key aspect of the mechanism of action of N-BPs that may have significant clinical relevance.

Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation.

We recently found that nucleosomes directly block access of CRISPR/Cas9 to DNA (Horlbeck et al., 2016). Here, we build on this observation with a comprehensive algorithm that incorporates chromatin, position, and sequence features to accurately predict highly effective single guide RNAs (sgRNAs) for targeting nuclease-dead Cas9-mediated transcriptional repression (CRISPRi) and activation (CRISPRa). We use this algorithm to design next-generation genome-scale CRISPRi and CRISPRa libraries targeting human and mouse genomes. A CRISPRi screen for essential genes in K562 cells demonstrates that the large majority of sgRNAs are highly active. We also find CRISPRi does not exhibit any detectable non-specific toxicity recently observed with CRISPR nuclease approaches. Precision-recall analysis shows that we detect over 90% of essential genes with minimal false positives using a compact 5 sgRNA/gene library. Our results establish CRISPRi and CRISPRa as premier tools for loss- or gain-of-function studies and provide a general strategy for identifying Cas9 target sites.

Smyd1 facilitates heart development by antagonizing oxidative and ER stress responses.

Smyd1/Bop is an evolutionary conserved histone methyltransferase previously shown by conventional knockout to be critical for embryonic heart development. To further explore the mechanism(s) in a cell autonomous context, we conditionally ablated Smyd1 in the first and second heart fields of mice using a knock-in (KI) Nkx2.5-cre driver. Robust deletion of floxed-Smyd1 in cardiomyocytes and the outflow tract (OFT) resulted in embryonic lethality at E9.5, truncation of the OFT and right ventricle, and additional defects consistent with impaired expansion and proliferation of the second heart field (SHF). Using a transgenic (Tg) Nkx2.5-cre driver previously shown to not delete in the SHF and OFT, early embryonic lethality was bypassed and both ventricular chambers were formed; however, reduced cardiomyocyte proliferation and other heart defects resulted in later embryonic death at E11.5-12.5. Proliferative impairment prior to both early and mid-gestational lethality was accompanied by dysregulation of transcripts critical for endoplasmic reticulum (ER) stress. Mid-gestational death was also associated with impairment of oxidative stress defense-a phenotype highly similar to the previously characterized knockout of the Smyd1-interacting transcription factor, skNAC. We describe a potential feedback mechanism in which the stress response factor Tribbles3/TRB3, when directly methylated by Smyd1, acts as a co-repressor of Smyd1-mediated transcription. Our findings suggest that Smyd1 is required for maintaining cardiomyocyte proliferation at minimally two different embryonic heart developmental stages, and its loss leads to linked stress responses that signal ensuing lethality.

Public attitude and knowledge on a new health policy for pharmaceutical care in Korea.

OBJECTIVE: to assess college student's attitude and knowledge of the 'separation of dispensing and prescribing' policy in Korea. DESIGN: a self-administered questionnaire survey of 700 college students. MAIN OUTCOME MEASURES: the attitude was assessed by the degree of interest in the policy, agreement to the policy need, expectation for the policy effect, and perceptions of motivation for physician's strike. The knowledge level was measured using four questions describing the goal/motivation of the policy and eight describing its operational rules. RESULTS: the level of interest (2.60 on a four-point scale), and agreement to the need (2.66) and the potential effect of the policy (2.29-2.91) were not very high. Concern for economic loss was perceived as the strongest motivation for physician's strike. While relatively well understood for the goals/motivations of the policy (mean score: 69.58 out of 100), the operational details of the policy were not well-informed (32.52). Interest and agreement with the policy need were the most significant factors affecting the knowledge level (P<0.01). CONCLUSION: For other public policies in the future, policy makers in Korea need to ensure public consent for the necessity of the policy and to develop more effective strategies to inform the public of the practical details of the policy.

[The emergence of Korean modern hospitals: hospitals in the late period of Chosun Dynasty].

Hospitals are confronting in the transforming or reforming period to cope with the rapid social and environmental changes worldwide. By the researches in the history of Korean health, we could understand the context of the introduction of Western medicine and institutions to Korea. However there have been few studies on the historical review of hospitals in relations to their roles in the modern medicine. This article is to review the issues around the rise of modern hospitals in Korean history of health affairs. The introduction of Western medicine in Korea was on the road with the establishment of Kwanghyewon, the Royal Hospital, which was possible due to favorable conditions under the Korean socio-political background for the emerging and accepting the entirely new medical system. And also the emergence of modern and transformed the Korean traditional health system from the fundamentals through the corruption of the old dynasty to nowadays. Most national health affairs including medical services, prevention of diseases, health promotion, and the training of health personnels have performed along with the development of modern hospitals, which have the roots in the period after the end of 19th century. Thus the Korean history of health care around the end of 19th century and beginning of 20th century could be defined as a period of emergence of modern hospitals. The hospitals also have played core roles in establishing the Korean modern health system and culture. Compared to the cases of Western countries, Korean modern hospitals were emerged with the exogenous factors in the turbulence of political and cultural changes in the world system. In sum, Korean modern hospitals in the period of late Chosun have the great meaning in that they are the beginning point to shape the current Korean health care system and the driving forces or carriers of this new system.

Partially penetrant postnatal lethality of an epithelial specific MicroRNA in a mouse knockout.

MicroRNAs are small noncoding RNAs thought to have pivotal roles in numerous diseases and developmental processes. However, a growing body of literature indicates that in vivo elimination of these tiny RNAs usually has little to no observable consequence, suggesting functional redundancy with other microRNAs or cellular pathways. We provide an in-depth analysis of miR-205 expression and define miR-205 as an epithelial-specific microRNA, and for the first time show that ablation of this microRNA knockout exhibits partially penetrant lethality in a constitutive mouse knockout model. Given the role of this microRNA in cancer and development, this mouse model will be an incredible reagent to study the function and mechanisms of miR-205 in epithelial tissue development and disease.

Association between total sleep duration and suicidal ideation among the Korean general adult population.

STUDY OBJECTIVES: Examine the association between sleep duration and suicidal ideation in Korean adults. DESIGN: Cross-sectional survey. SETTING: Data obtained by the Korea National Health and Nutrition Examination Survey IV (2007-2009) using a rolling sampling design involving a complex, stratified, multistage, and probability-cluster survey of civilian non-institutionalized Korean residents. PARTICIPANTS: A total of 15,236 subjects (6,638 males and 8,598 females) ≥ 19 years old. MEASUREMENTS AND RESULTS: The weighted prevalence of self-reported short sleep duration (≤ 5 h/day) was 11.7% in males and 15% in females, and of long sleep duration (≥ 9 h/day) was 6.7% in males and 8.9% in females. A U-shaped relationship existed, with both short and long sleep durations associated with a higher suicidal ideation risk. Multiple logistic regression analysis was used to analyze the relationship between sleep duration and suicidal ideation, adjusting for sociodemographic factors, health behavior, and health status. After controlling for covariates, people with short sleep were 38.1% more likely to have suicidal ideation (OR = 1.381, 95% CI 1.156-1.650) than people with sleep duration of 7 h/day. Suicidal ideation was 1.196 times higher (95% CI: 0.950-1.507) in long-sleeping people than people sleeping 7 h/day, although statistically not significant. Inclusion of depressive mood (a potential confounder) in multiple logistic regression models attenuated but did not eliminate the sleep duration/suicidal ideation association. LIMITATIONS: Sleep duration and suicidal ideation were assessed only by self-report. CONCLUSIONS: The sleep duration/suicidal ideation relationship is U-shaped in the Korean adult population. Self-reported habitual sleep duration may be a useful behavioral indicator for both individual and societal suicidal ideation risk.

Eps15R is required for bone morphogenetic protein signalling and differentially compartmentalizes with Smad proteins.

Transforming growth factor ß superfamily members signal through Smad transcription factors. Bone morphogenetic proteins (BMPs) act via Smads 1, 5 and 8 and TGF-ßs signal through Smads 2 and 3. The endocytic adaptor protein Eps15R, or 'epidermal growth factor (EGF) receptor pathway substrate 15-related protein' is a component of EGF signal transduction, mediating internalization of the EGF receptor. We show that it interacts with Smad proteins, is required for BMP signalling in animal caps and stimulates Smad1 transcriptional activity. This function resides in the Asp-Pro-Phe motif-enriched 'DPF domain' of Eps15R, which activates transcription and antagonizes Smad2 signalling. In living cells, Eps15R segregates into spatially distinct regions with different Smads, indicating an unrecognized level of Smad compartmentalization.

A resource for the conditional ablation of microRNAs in the mouse.

The importance of miRNAs during development and disease processes is well established. However, most studies have been done in cells or with patient tissues, and therefore the physiological roles of miRNAs are not well understood. To unravel in vivo functions of miRNAs, we have generated conditional, reporter-tagged knockout-first mice for numerous evolutionarily conserved miRNAs. Here, we report the generation of 162 miRNA targeting vectors, 64 targeted ES cell lines, and 46 germline-transmitted miRNA knockout mice. In vivo lacZ reporter analysis in 18 lines revealed highly tissue-specific expression patterns and their miRNA expression profiling matched closely with published expression data. Most miRNA knockout mice tested were viable, supporting a mechanism by which miRNAs act redundantly with other miRNAs or other pathways. These data and collection of resources will be of value for the in vivo dissection of miRNA functions in mouse models.

Tumor necrosis factor-receptor-associated factor-4 is a positive regulator of transforming growth factor-beta signaling that affects neural crest formation.

The transforming growth factor (TGF)-beta superfamily regulates cell proliferation, apoptosis, differentiation, migration, and development. Canonical TGFbeta signals are transduced to the nucleus via Smads in both major signaling branches, bone morphogenetic protein (BMP) or Activin/Nodal/TGFbeta. Smurf ubiquitin (Ub) ligases attenuate these pathways by targeting Smads and other signaling components for degradation by the 26S proteasome. Here, we identify tumor necrosis factor (TNF)-receptor-associated factor-4 (TRAF4) as a new target of Smurf1, which polyubiquitylates TRAF4 to trigger its proteasomal destruction. Unlike other TRAF family members, which mediate signal transduction by TNF, interleukin, or Toll-like receptors, we find that TRAF4 potentiates BMP and Nodal signaling. In the frog Xenopus laevis, TRAF4 mRNA is stored maternally in the egg animal pole, and in the embryo it is expressed in the gastrula marginal zone, neural plate, and cranial and trunk neural crest. Knockdown of embryonic TRAF4 impairs signaling, neural crest development and neural folding, whereas TRAF4 overexpression boosts signaling and expands the neural crest. In human embryonic kidney 293 cells, small interfering RNA knockdown of Smurf1 elevates TRAF4 levels, indicating endogenous regulation of TRAF4 by Smurf1. Our results uncover new functions for TRAF4 as a Smurf1-regulated mediator of BMP and Nodal signaling that are essential for neural crest development and neural plate morphogenesis.

Economic cost of dementia patients according to the limitation of the activities of daily living in Korea.

BACKGROUND: Dementia is expected to become a significant social burden in the future. However, there are few reports that have estimated the total economic cost of dementia, particularly according to the limitation in the activities of daily living (ADL) in Korea. OBJECTIVES: This study is to analyze the health care expenditures and cost per dementia patient, and estimated the total economic cost of dementia. METHODS: Caregivers of 609 dementia patients, who were randomly selected from a nationwide claim database of the Korean National Health Insurance Corporation, were interviewed using structured questionnaire in the period of September 2005. The total cost including the direct and indirect costs during a year was calculated. The patients were stratified into three limitation groups according to their ADL score. The cost according to the three limitation groups was analyzed. The national cost of dementia patients was then estimated. RESULTS: The total cost per dementia patient in Korea was $7,462. The direct and indirect costs were $6,626 and $836 per patient, respectively. The cost increased with increasing degree of limitation in the patient's ADL. Over a 1-year period, the total cost per patient in the low limitation group was lower ($3,698) than that of the moderate ($6,064) and high ($11,428) limitation group. CONCLUSIONS: This study demonstrates that the direct and indirect costs of dementia are considerately small for patients with lower limitation in their ADL. The total economic cost of dementia per year was estimated to be in the range of 1.3 to 3.3 billion dollars on assumptions in Korea.