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The fluorescence excimer emission of 1 is found to be enhanced upon the addition of K+ ion which is due to a conformational change of the ligand to have a more efficient π-π* interaction of two pyrenes. By contrast, no fluorescence change of 1 was observed in the anion binding test with F- , Cl- , Br- , I- , [image omitted] , [image omitted] , and [image omitted] as their tetrabutylammonium salts in MeCN. Interestingly, it was observed that the excimer emission of the two pyrenes decreases upon the addition of [image omitted] or [image omitted] in the 1√K+ complex solution. This is presumably due to a conformational change of the ligand by K+ ion binding, which induces the better conformation of two facing pyrene amides to selectively adopt [image omitted] or [image omitted] ion. This allosterical anion complexation could be applicable for NOR and INHIBIT (INH) logic gates. [ABSTRACT FROM AUTHOR]

This study aims to develop an optimized method for cryopreserving the germ cells of Lissachatina fulica ( L. fulica ) using vitrification, as an alternative approach for conserving endangered snail species. First, we isolated several key reproductive organs, including the sperm oviduct, albumen gland, hermaphrodite gland (ovotestis), and hermaphrodite duct from L. fulica . When the ovotestis was finely chopped, numerous sperm with long tails and distinct heads were observed. The staining of sperm nuclei was confirmed using Hoechst 33342 dye. Since the hermaphrodite gland, referred to as the ovotestis, contains both male and female germ cells, we performed tissue staining on the ovotestis using hematoxylin and eosin (H&E) dye. H&E staining of the ovotestis revealed numerous oval-shaped acini containing sperm and early germ cells. Spermatocytes and spermatids were observed within distinct boundaries, with mature sperm appearing following spermatogenesis. To preserve the species of the L. fulica , we introduced vitrification technology to cryopreserve its reproductive organs. The non-vitrification group showed an average cell viability of 96.6%, while the vitrification group had 86.8% after thawing. This study presents a reliable cryopreservation protocol for L. fulica , with potential applications for other endangered snails, supporting conservation efforts to preserve genetic resources and biodiversity.

This study is the first to identify bovine blastocysts through in vitro fertilization (IVF) of matured oocytes with a large quantity of high-quality sperm separated from a biomimetic cervix environment. We obtained high-quality sperm in large quantities using an IVF sperm sorting chip (SSC), which could mimic the viscous environment of the bovine cervix during ovulation and facilitates isolation of progressively motile sperm from semen. The viscous environment-on-a-chip was realized by formulating and implementing polyvinylpyrrolidone (PVP)-based solutions for the SSC medium. Sperm separated from the IVF-SSC containing PVP 1.5% showed high motility, normal morphology and high DNA integrity. As a result of IVF, a higher rate of hatching blastocysts, which is the pre-implantation stage, were observed, compared to the conventional swim-up method. Our results may significantly contribute to improving livestock with superior male and female genetic traits, thus overcoming the limitation of artificial insemination based on the superior genetic traits of existing males.

An increasing number of patients are receiving rehabilitation after stroke. But the impact of intensive rehabilitation on the long-term prognosis of patients with stroke remains to be elucidated. The purpose of this study was to identify the impact of intensive rehabilitation on the long-term prognosis of patients with stroke using data from the National Health Insurance Service database. This is a register-based, retrospective cohort study. Using data from the National Health Insurance Service database, we included the patients who received rehabilitation for stroke from 2006 to 2013. Of the 14,984 patients diagnosed with stroke, 2483 died within 1 year, and 2866 did not receive rehabilitation; hence, they were also excluded. The final sample included 9635 (49.2% men, 50.8% women) patients. After correcting for covariates, the Cox model was used to evaluate the effects of physical therapy (PT) and occupational therapy (OT) on survival. We estimated the independent contribution of each factor to the risk of death from the initiation of rehabilitation. Significant differences in mortality were observed according to age, Charlson comorbidity index (CCI), income level, and stroke type. Patients with stroke who received both PT and OT had a better long-term prognosis than those who received either treatment alone. Therapy performed by a physical therapist with more than 120 hours of training effectively improved the patients' long-term prognosis. Intensive PT and OT will help improve the long-term prognosis of patients with stroke. This study emphasizes the importance of intensive rehabilitation in these patients., Competing Interests: The authors have no funding and conflicts of interest to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Background: Large-scale epidemiological studies on pancreatic cancer in non-Western populations are insufficient. We investigated the risk factors for pancreatic cancer., Methods: Using the Korean National Health Insurance database, subjects who participated in the health examination program between 2005 and 2006 were identified and followed up until 2017. Adjusted hazard ratios (HRs) for pancreatic cancer risk were estimated using a Cox proportional hazards model., Results: During 11.5 years follow-up, 22,543 of 7,445,947 participants were newly diagnosed with pancreatic cancer. Compared with normal-weight subjects, pancreatic cancer risk was increased in those with severe obesity (BMI ≥ 28 kg/m 2 ) (HR = 1.16; 95% CI, 1.11-1.23). Subjects with diabetes had an increased risk compared with those without diabetes (HR = 1.48; 95% CI, 1.43-1.53). Current smokers had a higher risk than never smokers (HR = 1.43; 95% CI, 1.38-1.48). Current smoking combined with diabetes increased the risk compared with never smokers without diabetes (HR = 2.13; 95% CI, 2.00-2.28). Current smoking combined with BMI ≥ 25 kg/m 2 had an increased risk compared with never smokers with BMI < 23 kg/m 2 (HR = 1.55; 95% CI, 1.46-1.65)., Conclusion: Smoking, obesity, and diabetes are significant risk factors for pancreatic cancer in Koreans. Lifestyle modifications for smoking and obesity would be beneficial for pancreatic cancer prevention., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Cancer stem cells (CSCs) are a tumor cell subpopulation that drives tumor progression and metastasis, leading to a poor overall survival of patients. In colorectal cancer (CRC), the hyper-activation of Wnt/β-catenin signaling by a mutation of both adenomatous polyposis coli (APC) and K-Ras increases the size of the CSC population. We previously showed that CPD0857 inactivates Wnt/β-catenin signaling by promoting the ubiquitin-dependent proteasomal degradation of β-catenin and Ras proteins, thereby decreasing proliferation and increasing the apoptosis of CRC lines. CPD0857 also decreased the growth and invasiveness of CRC cells harboring mutant K-Ras resistant to EGFR mAb therapy. Here, we show that CPD0857 treatment decreases proliferation and increases the neuronal differentiation of neural progenitor cells (NPCs). CDP0857 effectively reduced the expression of CSC markers and suppressed self-renewal capacity. CPD0857 treatment also inhibited the proliferation and expression of CSC markers in D- K-Ras MT cells carrying K-Ras , APC and PI3K mutations, indicating the inhibition of PI3K/AKT signaling. Moreover, CPD0857-treated xenograft mice showed a regression of tumor growth and decreased numbers of CSCs in tumors. We conclude that CPD0857 could serve as the basis of a drug development strategy targeting CSCs activated through Wnt/β-catenin-Ras MAPK-PI3K/AKT signaling in CRCs.

Organoid cryopreservation method is one of key step in the organoid culture. We aimed to establish a simple and efficient cryopreservation method for mouse small intestinal organoids (MIOs) and colon organoids (MCOs) using various concentrations of cryoprotectant. Based on the theoretical simulation, we optimized the dimethyl sulfoxide (DMSO) concentration by pretreating the organoids with 5, 7.5, and 10% DMSO for 30 min at 4 °C to allow penetration into the organoids and evaluated their viability, proliferation, and function after cryopreservation. Gene expression in the MIOs and staining of lineage markers were examined real-time PCR. The organoids in the DMSO-treated groups as well as the control, expressed ChrgA, Ecad, Muc2, Lyz, villin, and Lgr5, and there are no significant. A forskolin-induced swelling assay for MIOs was performed to confirm normal cystic fibrosis transmembrane conductance regulator (CFTR) activity. Similar forskolin-induced swelling was observed in the DMSO-treated groups and the control. In addition, MCOs were transplanted into mouse colon for confirmation of regeneration therapy efficacy. Thawing organoids were cultured for two and four sequential passages after cryopreservation with 5% DMSO to confirm any changes in the gene expression of lineage markers after subculture. We developed a simple and efficient organoid freezing method using 5% DMSO with low potential toxicity and validated our findings with theoretical simulation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Inc.)

When ejaculated sperm travels through the vagina to the uterus, mucus secreted by the cervical canal generally filters out sperm having low motility and poor morphology. To investigate this selection principle in vivo, we developed a microfluidic sperm-sorting chip with a viscous medium (polyvinylpyrrolidone: PVP) to imitate the biophysical environment mimic system of the human cervical canal. The material property of the PVP solution was tuned to the range of viscosities of cervical mucus using micro-viscometry. The selection of high-quality human sperm was experimentally evaluated in vitro and theoretically analyzed by the convection-diffusion mechanism. The convection flow is shown to be dominant at low viscosity of the medium used in the sperm-sorting chip when seeded with raw semen; hence, the raw semen containing sperm and debris convectively flow together with suppressed relative dispersions. Also, it was observed that the sperm selected via the chip not only had high motilities but also normal morphologies and high DNA integrity. Therefore, the biomimetic sperm-sorting chip with PVP medium is expected to improve male fertility by enabling the selection of high-quality sperm as well as uncovering pathways and regulatory mechanisms involved in sperm transport through the female reproductive tract for egg fertilization.

Purpose: The study aimed to investigate the current status and prognostic factors for overall survival in patients who had undergone pulmonary metastasectomy for colorectal cancer., Materials and Methods: The data of 2,573 patients who had undergone pulmonary metastasectomy after surgery for colorectal cancer between January 2009 and December 2014 were extracted from the Korean National Health Insurance Service claims database. Patient-, colorectal cancer-, pulmonary metastasis-, and hospital-related factors were analyzed using the Kaplan-Meier method, log-rank test, and Cox proportional hazards analysis to identify prognostic factors for overall survival after pulmonary metastasectomy., Results: The mean age of the patients was 60.9±10.5 years; 66.2% and 79.1% of the participants were male and had distally located colorectal cancer, respectively. Wedge resection (71.7%) was the most frequent extent of pulmonary resection; 21.8% of the patients underwent repeated pulmonary metastasectomies; 73% of pulmonary metastasectomy cases were performed in tertiary hospitals; 53.9% of patients were treated in Seoul area; 82% of patients received chemotherapy in conjunction with pulmonary metastasectomy. The median survival duration was 51.8 months. The 3- and 5-year overall survival rates were 67.7% and 39.4%, respectively. In multivariate analysis, female sex, distally located colorectal cancer, pulmonary metastasectomy-only treatment, and high hospital volume (> 10 pulmonary metastasectomy cases/yr) were positive prognostic factors for survival., Conclusion: Pulmonary metastasectomy seemed to provide long-term survival of patients with colorectal cancer. The female sex, presence of distally located colorectal cancer, and performance of pulmonary metastasectomy in high-volume centers were positive prognostic factors for survival.

In vitro spermatogenesis can be performed for marine medaka (Oryzias dancena) via whole testis organ cultures, but spermatogenesis could only be maintained during the early phase of culturing, suggesting that the culture conditions can be further optimized. To improve the culture conditions, we examined the effects of culture temperature, basal media, and medium supplements on spermatogonial proliferation levels during whole testis organ culturing by BrdU incorporation assays. Our results show that a 30°C culture temperature negatively affected spermatogonial proliferation compared to 26°C and 28°C and that the use of Dulbecco's Modified Eagle Medium and Minimum Essential Medium α (α-MEM) was more effective for spermatogonial proliferation than the use of Leibovitz's L-15 Medium (L15). When fetal bovine serum (FBS) was replaced with KnockOut Serum Replacement (KSR), a significantly positive effect was observed for the maintenance of spermatogonial proliferation. However, supplementation of the medium with 17α, 20β-dihydroxy-4-pregnen-3-one did not show any significant effect. Gene expression analyses of four genes, including Nanos2, SCP3, AMH, and StAR, indicated that the optimized culture conditions consisting of α-MEM and KSR had the most positive influence on the maintenance of spermatogonial proliferation levels in whole testis organ cultures compared to the original culture conditions consisting of L15 and FBS by maintaining the function of Sertoli and Leydig cells. The results from this study will provide useful information for the study of in vitro spermatogenesis in fish., (© 2021. The Society for In Vitro Biology.)

This study aimed to identify the association between moving to a high-volume hospital and the mortality of patients with cancer living in the district. The study population comprised participants diagnosed with cancer within the past nine years (2004-2012). The final sample included 8197 patients with cancer, 3939 were males (48.1%), and 4258 were females (51.9%). A Cox proportional hazard model was used to estimate the hazard ratio (HR) for death. Confounding variables including sex, age, type of social security, income level, disability, and utilization volume were incorporated into the model. Among patients with cancer living in the district, 2874 (35.1%) used healthcare services in Seoul. About 10% ( n = 834) of patients died during the follow-up period. The HR for death in females (HR: 0.68, 95% CI: 0.58-0.81) was lower than that in males. Additionally, the HR for the death of patients using healthcare services in Seoul (HR: 1.30, 95% CI: 1.11-1.53) was higher than those patients who did not use healthcare services in Seoul. Among patients utilizing services in the province, wealthier patients' survival probability was significantly higher than that of others. The cause of income differences should be identified, and accessibility to medical use of low-income families should be enhanced to prevent mortality of patients from cancer disparities.

Introduction  The association between tonsillectomy with adenoidectomy (T&A) with appendicitis is controversial, and the association of T&A with pneumonia has not been investigated. Objective  To investigate the associations of T&A with pneumonia and appendicitis using data from the Korean National Health Insurance Service National Sample Cohort. Methods  We selected patients between the ages of 3 and 10 years who had undergone T&A in 2005 and were monitored since the performance of the T&A until 2013. The control group was established to have similar propensities for demographic characteristics compared to the T&A group. For eight years after the T&A, the number of patients with a diagnosis of pneumonia, patients who were admitted due to pneumonia, and those who underwent appendectomy were analyzed. The risk factors for pneumonia and appendectomy were analyzed. Results  The number of pneumonia diagnoses was significantly higher in the T&A group than in the control group ( p  = 0.023), but there were no significant differences in the number of admissions due to pneumonia between the 2 groups ( p  = 0.155). Younger age and T&A were significant risk factors for the development of pneumonia. There were no significant differences in the number of appendectomies between the T&A and the control groups ( p  = 0.425), neither were there significant risk factors for appendectomy. Conclusion  Tonsillectomy with adenoidectomy was associated with an increase in pneumonia diagnoses, but it was not associated with the number of appendectomies. The associations of T&A with pneumonia and appendicitis were analyzed in this population-based study., Competing Interests: Conflict of Interests The authors have no conflict of interests to declare., (Fundação Otorrinolaringologia. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commecial purposes, or adapted, remixed, transformed or built upon. ( https://creativecommons.org/licenses/by-nc-nd/4.0/ ).)

Background: The effect of tonsillectomy with adenoidectomy (T&A) on otitis media has been investigated, but there have been no reports of the relationship between T&A and medical services used in association with otitis media. We investigated the effect of T&A on otitis media with regard to the number and cost of medical services used., Methods: From the National Health Insurance Service National Sample Cohort data in Korea, we selected patients 7 years old or younger in 2002 who had T&A in 2005 while between the ages of three and ten. A control group was established matching the patient group with similar propensities of demographic characteristics. The number and cost of medical services used in association with otitis media were analyzed for 3 years before T&A through 8 years after T&A., Results: The total number of patients was 1,338, with 227 in the T&A group and 1,111 in the non-T&A group. The number of medical services used was not significantly different between the T&A and non-T&A groups before and after surgery. The cost of medical services used was significantly higher in the T&A group than in the non-T&A group one year before surgery. The cost of medical services used was not significantly different between the two groups after surgery., Conclusions: There were no significant differences between the T&A and non-T&A groups in the number and cost of medical services used in association with otitis media after surgery., Competing Interests: Competing interestsThe authors declare that they have no competing interests., (© The Author(s) 2020.)

Background: Oncogenic K-Ras mutations in colorectal cancer (CRC) combined with APC mutations worsen CRC prognosis and lower drug effectiveness. Thus, inhibition of both Wnt/β-catenin and Ras-MAPK signaling may be a rational strategy to improve the treatment of this cancer., Objective: To identify a novel compound inhibiting both Wnt/β-catenin and Ras-MAPK signaling in CRC., Methods and Patients: We developed a two-part screening system consisting of analysis of TOP flash reporter cells and then potential toxicity effects on primary neural stem cells (NSCs). We then screened 2000 chemical compounds and tested efficacy of candidates against isogenic colon cancer cells harboring wild-type or mutant K-Ras. We employed immunohistochemistry and immunocytochemistry to determine marker signatures associated with development of disease phenotypes., Results: We identified CPD0857, a compound that inactivates Wnt/β-catenin signaling and promotes ubiquitin-dependent proteasomal degradation of β-catenin and Ras proteins. CPD0857 effectively decreased proliferation and increased apoptosis of CRC cell lines, and overcame resistance of CRC harboring APC and K-Ras mutations to treatment with an EGFR monoclonal antibody (mAb). Moreover, CPD0857 attenuated invasiveness of highly migratory CRC cells in vitro. Accordingly, xenograft mice treated with CPD0857 showed slower tumor growth and significant decreases in both β-catenin and Ras protein expression., Conclusions: CPD0857 may be a potential drug for treating aggressive CRC carrying mutations that aberrantly activate Wnt/β-catenin and Ras-ERK pathways.

Introduction: Relatively few, mainly cross-sectional, studies have examined the relationship between smoking cessation and metabolic syndrome (MetS). In particular, information on smoking cessation after MetS is limited. This study aimed to investigate the probability of smoking cessation after the onset of MetS., Methods: In this study we used cohort data from a rural area of Korea and extracted the data of 1054 smokers who were identifiable at baseline and were followed up. Of these, 1041 individuals were selected. Descriptive statistical analyses were performed to identify the basic characteristics of smokers. Multiple logistic regression was performed to determine the association between changes in MetS and smoking cessation., Results: The probability of smoking cessation was 1.84 times higher in the newly developed MetS cohort than in the reference group (without MetS at any time point), and it was 1.61 times higher in the persistent MetS cohort than in the reference group, with both probabilities being significant., Conclusions: We found that patients with MetS were more likely to quit smoking than those without MetS. However, intervention is still needed, as numerous patients with MetS continued to smoke. Interventions that actively involve medical institutions or organizations are among the most effective approaches to promote smoking cessation in patients with MetS. In particular, women, farmers and current drinkers should be prioritized., Competing Interests: The authors have completed and submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest and none was reported., (© 2020 Park M.B. et al.)

Gd 3+ -based contrast agents monopolize in the clinical MR imaging-based diagnosis of hepatic tumors, however, the inherent toxicity by the released Gd 3+ -ions triggered an urgent demand for safer alternatives. Here, we demonstrate hollow manganese silicate nanoparticles (HMS), which exert burst-release of Mn 2+ -ions by switching to physiological acidic condition, exhibiting high effectiveness in hepatic tumor characterization as liver-specific MR contrast agent through the in-depth in vivo MR imaging study and immunohistochemical investigations with three hepatic tumor models (e.g., hepatocellular carcinoma, neuroendocrine carcinoma, adenocarcinoma). Their characteristic time-sequential enhancement patterns in HMS-enhanced MR imaging with improved conspicuity reflect their biological features such as vascularity, cellularity, mitochondrial activity and hepatocellular specificity, and thus allow the disease-specific characterization of various hepatic tumors. HMS-enhanced MR imaging with necrotic hepatocellular carcinoma model suggested the good correlation of the extent of tumor necrosis with residual mitochondrial activity. Such multi-responsive spatio-biological distribution and function of HMS resulting in time-dependent bioimaging coupled with low systemic toxicity sets the clinical potential to accurate diagnosis and therapeutic response in various hepatic tumors., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Objective: To evaluate whether the surface layer (S-layer) protein of Lactobacillus brevis serves as a self-aggregating protein tag for cost-effective separation of human and yeast D-amino acid oxidases (hDAAO and yDAAO) expressed in E. coli., Results: In aqueous two-phase (PEG-phosphate) system, the S-layer:DAAO fusion proteins (shDAAO and syDAAO) were separated at the interface with a recovery of 82 ± 10.6% for shDAAO and 95 ± 1.9% for syDAAO. Some shDAAO proteins were separated as precipitates with a recovery of 41 ± 0.5% in phosphate (9%, w/w) using PEG 3000 and PEG 4000 (16%, w/w), while some syDAAO proteins were also isolated as precipitates with a recovery of 75 ± 17.5% in phosphate (9%, w/w) using PEG 4000 and PEG 8000 (16%, w/w)., Conclusions: The S-layer of L. brevis was applied to a self-assembled protein tag to enable cost-effective separation of human and yeast D-amino acid oxidases expressed in E. coli cells. Because of the self-assembling properties of S-layer proteins, human and yeast D-amino acid oxidases fused with S-layer proteins could be easily separated by aggregates at the interface and/or in a few conditions by precipitates to the bottom of the PEG-phosphate aqueous system.

Purpose: This study investigated the association between health status and tooth loss based on data from the National Health Insurance Service-Health Examinee Cohort in 2002-2015., Methods: Multivariate Cox proportional hazards regression analyses were applied to a longitudinal retrospective database, which was updated and newly released in 2018, to assess the association between health status and tooth loss while adjusting for potential confounders among sociodemographic and economic factors (sex, age, household income, insurance, and presence of disability), general and oral health status (body mass index [BMI], smoking and drinking status, periodic dental visits and scaling, and brushing before sleep), and comorbid disease (hypertension, diabetes mellitus [DM], and Charlson comorbidity index [CCI])., Results: Among 514,866 participants from a South Korean population, 234,247 (45.5%) participants satisfying the inclusion criteria were analyzed. In the adjusted multivariate analysis, sex, age, household income, insurance, presence of disability, BMI, smoking and drinking status, periodic scaling, tooth brushing before sleep, DM, and CCI showed statistically significant associations with the loss of at least 1 tooth. The risk of experiencing a loss of ≥4 teeth was associated with an increase in age (in those 50-59 years of age: hazard ratio [HR], 1.98; 95% confidence interval [CI], 1.93-2.03; in those 60-69 years of age: HR, 2.93; 95% CI, 2.85-3.02; and in those 70-79 years of age: HR, 2.93; 95%, CI 2.81-3.05), smoking (HR, 1.69; 95% CI, 1.65-1.73), and DM (HR, 1.43; 95% CI, 1.38-1.48)., Conclusions: The results of this study showed that the risk of experiencing tooth loss was related to multiple determinants. DM and smoking were especially significantly associated with tooth loss., Competing Interests: Conflict of Interest: No potential conflict of interest relevant to this article was reported.

Competing Interests: The authors have no potential conflicts of interest to disclose.

Introduction: The goal of this study was to evaluate the differences in urine cotinine (UC) concentration based on the use of conventional cigarettes, e-cigarettes (ECs), and dual product use, and determine the use of ECs in the real world., Methods: In total, 15099 subjects were classified into non-smokers, cigarette smokers (c-smokers), e-cigarette smokers (e-smokers), and dual users, and their UC (a classical biomarker of smoking) values were compared. Analysis of covariance (ANCOVA) was performed after adjusting for age, sex and job status to analyze the differences in UC concentration in relation to type of smoking. The reasons for using ECs and the experience of cigarette use before using ECs were analyzed., Results: Of the 15099 people, 76.4% were non-smokers, 20.9% c-smokers, 2.3% dual users, and 0.4% were e-smokers. There were significant differences in UC concentration among the groups (p<0.001). The geometric mean (GM) UC concentration was 4.45 ng/mL. UC concentration was the highest among dual users (GM: 1030.5, median: 1258.9 ng/mL), followed by c-smokers (GM: 842.5, median: 1163.0 ng/mL), e-smokers (GM: 119.5, median: 309.7 ng/mL), and non-smokers (GM: 0.8, median: 0.8 ng/mL). Among the EC users, the rate of using ECs for health or social convenience was 81.9%. Among e-smokers, 11.4% had never smoked previously., Conclusions: The UC concentration was the highest among dual users. However, for the female population, the UC concentration was the highest among e-smokers. The vast majority of EC users were dual users. In addition, there were no differences in the frequency of cigarette smoking between the dual user and c-smoker groups. Consequently, EC use did not lead to a decrease in cigarette use, but did lead to an increase in UC concentration. Therefore, in the real world, dual users have higher cotinine levels than the other groups, which could indicate that they take more nicotine by cigarettes or ECs, or are more addicted than others., Competing Interests: Authors have completed and submitted the ICMJE Form for Disclosure of Potential Conflicts of Interest and none was reported., (© 2019 Park M. B.)

It is necessary to confirm the effect of long-term care insurance (LTCI) by identifying changes in mortality, whether benefits are used or not, as well as the effects of in-home and institutional services on mortality. The goal of this study was to identify the association between service use and the mortality rate in elderly Koreans. We used Cox proportional hazard regression models and the Kaplan-Meier survival curve method to estimate the hazard ratio and survival probability for death while adjusting for covariates. We detected a 27.8% mortality rate at the 40-month follow-up period. Male gender, advanced age and activities of daily living were risk factors for mortality. In all models, the hazard ratio of participant death of those using long-term care services was significantly lower than for those who did not use these services. Among the service users, the hazard ratio for participant death of institutional service users was significantly higher than it was for in-home service users. This study also identified the impact of the transition from in-home services to institutional services. A primary goal of LTCI is to promote health and life stabilization in the elderly. To both delay and prevent institutionalization, it is necessary to develop assistive devices and effective in-home services and ensure access of these for elderly patients., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Mammalian oocytes have a proteinaceous hydrogel-like outer shell known as the zona pellucida (ZP) that semi-encloses their plasma membrane and cytoplasm. In this study, we cryopreserved mouse oocytes either with or without ZP by vitrification. Our results show that the presence of an intact ZP could significantly improve the post-vitrification survival of oocytes to 92.1% from 13.3% for oocytes without ZP. Moreover, there was no significant difference in embryonic development between fresh and cryopreserved oocytes with ZP after in vitro fertilization (IVF). Further atomic force microscopy (AFM) analysis showed that the intact oocytes with ZP have an elastic modulus that is more than 85 times higher than that of oocytes without ZP. This may partially explain the important role of ZP in protecting the oocytes by resisting the mechanical stress due to possible ice formation during cryopreservation by vitrification. Collectively, this study reveals a new biophysical role of ZP during vitrification of oocytes and suggests microencapsulation of the many mammalian cells without a ZP in ZP-like hydrogel is an effective strategy to improve their survival post cryopreservation by vitrification., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Cryopreservation by low-cryoprotectant (CPA) vitrification has the potential to combine all the advantages of the conventional high-CPA vitrification and slow-freezing approaches while avoiding their drawbacks. However, current low-CPA vitrification protocol for cryopreservation of oocytes requires a lengthy and multi-step procedure for unloading CPAs. In this study, we report a much-simplified procedure of using quartz microcapillary (QMC) for low-CPA vitrification of mouse oocytes with only one step for unloading CPAs. The immediate viability of oocytes after the improved low-CPA vitrification was determined to be more than 90%. Moreover, no significant difference was observed in terms of embryonic development from the two-cell to blastocyst stages between the fresh and vitrified oocytes after in vitro fertilization (IVF). This improved low-CPA vitrification technology has the potential for efficient cryopreservation of oocytes to preserve the fertility of mammals including humans for assisted reproductive medicine, maintenance of animal resource and endangered species, and livestock management., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Contemporary systems for in vitro culture of ovarian follicles do not recapitulate the mechanical heterogeneity in mammalian ovary. Here we report microfluidic generation of biomimetic ovarian microtissue for miniaturized three-dimensional (3D) culture of early secondary preantral follicles by using alginate (harder) and collagen (softer) to fabricate the ovarian cortical and medullary tissues, respectively. This biomimetic configuration greatly facilitates follicle development to antral stage. Moreover, it enables in vitro ovulation of cumulus-oocyte complex (COC) from the antral follicles in the absence of luteinizing hormone (LH) and epidermal growth factor (EGF) that are well accepted to be responsible for ovulation in contemporary literature. These data reveal the crucial role of mechanical heterogeneity in the mammalian ovary in regulating follicle development and ovulation. The biomimetic ovarian microtissue and the microfluidic technology developed in this study are valuable for improving in vitro culture of follicles to preserve fertility and for understanding the mechanism of follicle development and ovulation to facilitate the search of cures to infertility due to ovarian disorders., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

We report the chiroptical signature and binding interactions of cationic (meso-tetrakis(4-N-methylptridyl)porphyrin, 2HT4) and anionic (meso-tetrakis(4-sulfonatophenyl)porphyrin, 2HTPPS) porphyrins and their zinc(II) and nickel(II) derivatives (ZnT4, ZnTPPS, NiT4, and NiTPPS) with right-handed B-form and two forms of left-handed Z-form of alternating guanine-cytosine polydeoxynucleotide poly(dG-dC)2. NiTPPS is able to spectroscopically discriminate between spermine-induced Z-DNA and Co(III)-induced Z-DNA via new induced circular dichroism signal in the visible region of the electromagnetic spectrum., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The left-handed Z-DNA form of the short unmodified alternating guanine-cytosine oligonucleotides, 5'-(dGdC)(24) and 5'-(dGdC)(18), was selectively detected under physiological ionic strength and pH conditions using the anionic nickel(II) porphyrin, NiTPPS. No spectroscopic signal was observed for NiTPPS with any right-handed oligonucleotides under identical conditions. The 48mer 5'-(dGdC)(24) Z-form was detected at concentrations as low as 100nM. The binding of NiTPPS to the B- and Z-oligonucleotides was studied quantitatively by UV-vis absorption and circular dichroism spectroscopies. NiTPPS was found to be a universal DNA binder, with binding affinity and geometry depending on the ionic composition of the solution, rather than on the DNA helical twist. This is the first example of a successful spectroscopic detection of the Z-DNA of short unmodified oligonucleotides under physiological pH and ionic strength conditions., (Copyright (c) 2010 Elsevier Inc. All rights reserved.)