Potassium channels in human skin fibroblast have been studied as a possible site of Alzheimer disease pathogenesis. Fibroblasts in Alzheimer disease show alterations in signal transduction pathway such as changes in Ca2+ homeostasis and/or Ca2+-activated kinases, phosphatidylinositol cascade, protein kinase C activity, cAMP levels and absence of specific K+ channel. However, little is known so far about electrophysiological and pharmacological characteristics of large-conductance Ca2+-activated K+ (BKCa) channel in human fibroblast (CRL-1474). In the present study, we found Iberiotoxin- and TEA-sensitive outward rectifying oscillatory current with whole-cell recordings. Single channel analysis showed large conductance K+ channels (106 pS of chord conductance at +40 mV in physiological K+ gradient). The 106 pS channels were activated by membrane potential and [Ca2+]i, consistent with the known properties of BKCa channels. BKCa channels in CRL-1474 were positively regulated by adenylate cyclase activator (10microM forskolin), 8-Br-cyclic AMP (300microM) or 8-Br-cyclic GMP (300microM). These results suggest that human skin fibroblasts (CR-1474) have typical BKCa channel and this channel could be modulated by c-AMP and c-GMP. The electrophysiological characteristics of fibroblasts might be used as the diagnostic clues for Alzheimer disease.
Adenylyl Cyclases ; Alzheimer Disease ; Fibroblasts* ; Homeostasis ; Humans ; Membrane Potentials ; Nucleotides, Cyclic* ; Patch-Clamp Techniques ; Phosphatidylinositols ; Phosphotransferases ; Potassium Channels ; Protein Kinase C ; Second Messenger Systems ; Signal Transduction ; Skin*

Hydrogen peroxide (H2O2) causes oxidative stress and is considered as an inducer of cell death in various tissues. Two-pore domain K+ (K2p) channels may mediate K+ efflux during apoptotic volume decreases (AVD) in zygotes and in mouse embryos. In the present study, we sought to elucidate linkage between K2p channels and cell death by H2O2. Thus K2p channels (TASK-1, TASK-3, TREK-1, TREK-2) were stably transfected in HEK-293 cells, and cytotoxicity assay was preformed using cell counting kit-8 (CCK-8). Cell survival rates were calculated using the cytotoxicity assay data and dose-response curve was fitted to the H2O2 concentration. Ionic currents were recorded in cell-attached mode. The bath solution was the normal Ringer solution and the pipette solution was high K+ solution. In HEK-293 cells expressing TREK-1, TREK-2, TASK-3, H2O2 induced cell death did not change in comparison to non-transfected HEK-293. In HEK-293 cells expressing TASK-1, however, dose-response curve was significantly shifted to the left. It means that H2O2 induced cell death was increased. In cell attached-mode recording, application of H2O2 (300micrometer) increased activity of all K2P channels. However, a low concentration of H2O2 (50micrometer) increased only TASK-1 channel activity. These results indicate that TASK-1 might participate in K+ efflux by H2O2 at low concentration, thereby inducing AVD.
Animals ; Apoptosis* ; Baths ; Cell Count ; Cell Death ; Cell Survival ; Embryonic Structures ; Hydrogen Peroxide* ; Hydrogen* ; Mice ; Oxidative Stress ; Zygote

Purpose: This study aimed to identify the job characteristics of the integrated nursing care wards. Methods: For 388 nurses working in the integrated nursing wards of 30 hospitals, the importance, performance frequency, and difficulty of nursing tasks were analyzed using 31 job categories (678 items). Nursing tasks were analyzed using ImportancePerformance Analysis by hospital type. Results: Tertiary hospitals and general hospitals were analyzed using Importance-Performance Analysis, and the categories of general nursing intervention, spiritual and end of life nursing, and nursing during examination differed by hospital type. Other tasks into the same categories. 'Keep up the Good Work' includes 12 tasks: nursing assessment, medication and blood transfusion, admission or discharge management, and cooperation and coordination. 'Concentrate Here' had three tasks: emergency care, education to nurses, self-development. 'Possible Overkill' included supportive contact, environmental management, and product management. 'Low Priority' has ten tasks, including administration and organization management, facility management. Conclusion Nurses had different perceptions of importance and difficulty according to the tasks.Nursing during the examination, general nursing intervention, spiritual nursing, and end-of-life nursing were placed in different domains according to hospital type. Therefore, it is necessary to establish the nursing tasks and plan to improve workforce management, reflecting these differences.

Purpose: The purpose of this study was to identify factors influencing job satisfaction among ambulatory care nurses. Methods: Data were collected through a self-evaluation survey of 129 nurses in one tertiary hospital. Data were analyzed using paired t-test, independent t-test, one-way ANOVA, Importance-Performance Analysis, Pearson correlation coefficients, and multiple regression analysis. Results: The mean scores were: for importance, 3.18 out of 4, for performance, 2.24 out of 4, for professional self-concept, 2.92 out of 4, and for job satisfaction 2.77 out of 5. There were statistically significant positive correlations between professional self-concept and job satisfaction (r=.37, p<.001). Stepwise multiple regression analysis showed that satisfaction with professional self-concept, clinical career, and age explained 47.0% of the job satisfaction of ambulatory care nurses (F=37.51, p<.001). Satisfaction with professional self-concept, clinical career, and age were statistically significant predictors of job satisfaction. Conclusion Findings indicate motivation programs according to clinical career would be useful to improve job satisfaction of ambulatory care nurses. Additionally, continuous education should be provided to strengthen the professional self-concept of nursing professionals in both undergraduate curricula and clinical fields.

Exogenous carbon monoxide (0.2%) increases L-type calcium (Ca2+) current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type Ca2+ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type Ca2+ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type Ca2+ current was partially blocked by N- (3- (Amino-methyl) benzyl) acetamidine-2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or N5- (1-Iminoethyl)-L-ornithine dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type Ca2+ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.
Barium ; Calcium Channels, L-Type ; Calcium* ; Carbon Monoxide ; Carbon* ; Hand ; Humans* ; Membranes ; Muscle, Smooth* ; Myocytes, Smooth Muscle* ; Nitric Oxide Synthase* ; Nitric Oxide* ; Protein Isoforms

Polyploidization is a process by which cells are induced to possess more than two sets of chromosomes. Although polyploidization is not frequent in mammals, it is closely associated with development and differentiation of specific tissues and organs. The liver is one of the mammalian organs that displays ploidy dynamics in physiological homeostasis during its development. The ratio of polyploid hepatocytes increases significantly in response to hepatic injury from aging, viral infection, iron overload, surgical resection, or metabolic overload, such as that from non-alcoholic fatty liver diseases (NAFLDs). One of the unique features of NAFLD is the marked heterogeneity of hepatocyte nuclear size, which is strongly associated with an adverse liver-related outcome, such as hepatocellular carcinoma, liver transplantation, and liver-related death. Thus, hepatic polyploidization has been suggested as a potential driver in the progression of NAFLDs that are involved in the control of the multiple pathogenicity of the diseases. However, the importance of polyploidy in diverse pathophysiological contexts remains elusive. Recently, several studies reported successful improvement of symptoms of NAFLDs by reducing pathological polyploidy or by controlling cell cycle progression in animal models, suggesting that better understanding the mechanisms of pathological hepatic polyploidy may provide insights into the treatment of hepatic disorders.

The concept of cognitive frailty has recently been proposed by an International Consensus Group as the presence of physical frailty and cognitive impairment [defined using the Clinical Dementia Ratings (CDR)=0.5], without concurrent dementia. However, CDR is difficult to implement and not often available in epidemiologic studies or busy clinical settings, and an alternative to CDR is required. We suggest an alternative definition of cognitive frailty as: 1) physical frailty, 2) more than 1.5 standard deviation below the mean for age-, gender-, and education-adjusted norms on any cognitive function test (e.g., the Montreal Cognitive assessment test, the Alzheimer’s disease assessment scale-cognitive subscale, verbal learning test, Digit Span, Boston Naming Test, Trail Making Test, and Frontal Assessment Battery), and 3) no dependency in instrumental activities of daily living. The redefined criteria for cognitive frailty would be more feasible to implement and thus more applicable in epidemiologic studies and busy clinical settings.
Activities of Daily Living ; Cognition ; Cognition Disorders ; Consensus ; Dementia ; Epidemiologic Studies ; Mild Cognitive Impairment ; Trail Making Test ; Verbal Learning

BACKGROUND: The plasma levels of cell-free DNA (cfDNA) are known to be elevated under inflammatory or apoptotic conditions. Increased cfDNA levels have been reported in hemodialysis (HD) patients. The aim of this study was to investigate the clinical significance of cfDNA in HD patients. METHODS: A total of 95 patients on HD were enrolled. We measured their predialysis cfDNA levels using real-time EIF2C1 gene sequence amplification and analyzed its association with certain clinical parameters. RESULTS: The mean plasma cfDNA level in the HD patients was 3,884 +/- 407 GE/mL, and the mean plasma cfDNA level in the control group was 1,420 +/- 121 GE/mL (P < 0.05). Diabetic patients showed higher plasma cfDNA levels compared with nondiabetic patients (P < 0.01). Patients with cardiovascular complications also showed higher plasma cfDNA levels compared with those without cardiovascular complication (P < 0.05). In univariable analysis, the cfDNA level was associated with 3-month mean systolic blood pressure (SBP), white blood cell, serum albumin, creatinine (Cr), normalized protein catabolic rate in HD patients. In diabetic patients, it was significantly correlated with SBP, hemoglobin A1c, and serum albumin. In multivariate analysis, SBP was the independent determinant for the cfDNA level. In diabetic patients, cfDNA level was independently associated with hemoglobin A1c and SBP. CONCLUSIONS: In patients with HD, cfDNA is elevated in diabetic patients and patients with cardiovascular diseases. Uncontrolled hypertension and poor glycemic control are independent determinants for the elevated cfDNA. Our data suggest that cfDNA might be a marker of vascular injury rather than proinflammatory condition in HD patients.
Blood Pressure* ; Cardiovascular Diseases ; Creatinine ; Diabetes Mellitus ; DNA* ; Humans ; Hypertension ; Leukocytes ; Multivariate Analysis ; Plasma* ; Renal Dialysis* ; Serum Albumin ; Vascular System Injuries

Pruritus (itching) is classically defined as an unpleasant cutaneous sensation that leads to scratching behavior. Although the scientific criteria of classification for pruritic diseases are not clear, it can be divided as acute or chronic by duration of symptoms. In this study, we investigated whether skin injury caused by chemical (contact hypersensitivity, CHS) or physical (skin-scratching stimulation, SSS) stimuli causes initial pruritus and analyzed gene expression profiles systemically to determine how changes in skin gene expression in the affected area are related to itching. In both CHS and SSS, we ranked the Gene Ontology Biological Process terms that are generally associated with changes. The factors associated with upregulation were keratinization, inflammatory response and neutrophil chemotaxis. The Kyoto Encyclopedia of Genes and Genomes pathway shows the difference of immune system, cell growth and death, signaling molecules and interactions, and signal transduction pathways. Il1a , Il1b and Il22 were upregulated in the CHS, and Tnf, Tnfrsf1b, Il1b, Il1r1 and Il6 were upregulated in the SSS. Trpc1 channel genes were observed in representative itching-related candidate genes. By comparing and analyzing RNA-sequencing data obtained from the skin tissue of each animal model in these characteristic stages, it is possible to find useful diagnostic markers for the treatment of itching, to diagnose itching causes and to apply customized treatment.
Animals ; Biological Processes ; Chemotaxis ; Classification ; Cytokines ; Dermatitis, Contact* ; Gene Expression ; Gene Ontology ; Genome ; Hypersensitivity ; Immune System ; Interleukin-6 ; Mice* ; Models, Animal ; Neutrophils ; Pruritus* ; RNA* ; Sensation ; Sequence Analysis, RNA* ; Signal Transduction ; Skin* ; Transcriptome ; Transient Receptor Potential Channels ; Up-Regulation ; Wound Healing

Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipient's plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patient's clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.
Biopsy ; Creatinine ; DNA ; Humans ; Kidney Transplantation* ; Kidney* ; Monitoring, Physiologic ; Plasma ; Polymerase Chain Reaction* ; Polymorphism, Single Nucleotide ; Tissue Donors ; Transplant Recipients ; Transplants