Cellular cardiomyoplasty has recently emerged as a potential new treatment of ischemic heart disease. Combining cellular cardiomyoplasty with gene therapy using myogenic transcription factor might facilitate myocardial regeneration. In this study, we engineered H9c2, L6 using plasmid vector to overexpress the transcription factor MEF2c, Nkx2.5 involved in cardiomyogenesis. We investigated 1) formation of intercellular junction in mono-culture and co-culture with cardiomyocyte for functional and structural synchronous contraction after transplantation, 2) differentiation into cardiomyocyte, 3) resistance to hypoxic condition. Each cell overexpressing MEF2 and Nkx2.5 was generated by gene transfection and clonal selection. CO-culture was performed that each cell line added over cultured cardiomyocyte. H9c2-MEF2c and H9c2-Nkx2.5 became long, spindle shape like cardiomyocyte. Troponin T, cardiac specific marker, was found spot-like pattern in H9c2-Nkx2.5. However, co-culture with cardiomyocyte did not induce differentiation all kinds of cells into cardiomyocyte. Connexin43, which is gap junction marker was increased in H9c2-MEF2c, H9c2-Nkx2.5, L6-MEF2c and L6-Nkx2.5. Especially, co-culture with cardiomyocyte resulted in elevation of connexin43 levels more than monoculture. Ultrastructurally, formations of gap junction and desmosome were found apparently in L6-Nkx2.5. Long-standing, strong, regular and more frequent contraction were observed in cardiomyocyte co-cultured with H9c2-MEF2c, H9c2-Nkx2.5, L6-MEF2c, L6-Nkx2.5, respectively. Neverthless, any cell did not have active contraction itself, but passive movement except cardiomyocyte. H9c2-MEF2c, L6-MEF2c and L6-Nkx2.5 had resistance to hypoxia compared with other groups. These results suggested that co-culture and overexpressions of MEF2c and Nkx2.5 induced differentiation into cardiomyocyte and played an important role on intercellular junction formation and hypoxic resistance. This would be a promising source of cellular cardiomyoplasty. Therefore, much more research would be essential for clinical application of cellular cardiomyoplasty and this study would be a basic source for further study of MEF2c and Nkx2.5 in cellular cardiomyoplasty.
Animals ; Anoxia ; Cardiomyoplasty ; Cell Line ; Cell Transplantation ; Coculture Techniques* ; Connexin 43 ; Desmosomes ; Gap Junctions ; Genetic Therapy ; Intercellular Junctions ; Myoblasts ; Myoblasts, Cardiac* ; Myocardial Ischemia ; Myocytes, Cardiac* ; Plasmids ; Rats* ; Regeneration ; Transcription Factors ; Transfection ; Troponin T

BACKGROUND AND OBJECTIVE: Even though they are identified on the basis of their induction by elevated temperatures, heat shock proteins are induced by various stimuli. Such inducers include viral infection, exposure to ethanol, sodium arsenite, steroid hormones and heavy metals. They become involved under pathological conditions, such as anoxia/ischemia, reperfusion injury and oxidative stress in the heart. In this research, the cellular damages and changes of Hsp25 and alphaB crytallin were investigated under oxidative stress induced H2O2 treatment in a cultured cardiomyoblast cell line, H9c2. MATERIALS AND METHODS: The morphological changes of the H9c2, and the distributions of Hsp25 and alphaB crytallin, were examined with phase-contrast, confocal and electron microscopy. The levels of Hsp25 and alphaB crytallin expressions within the cells were also checked. RESULTS: After exposure to a low concentration (10 micrometer) of H2O2, the amounts of Hsp25 and alphaB crytallin were increased more than in the control and heat shock groups. They were particularly localized in the perinuclear region and along the actin filament. When exposed to a high concentration (more than 100 micrometer) of H2O2, many cells developed apoptotic features, such as vacuolization, blebbing and apoptotic body formations. The amounts of Hsp25 and alphaB crytallin were decreased more than in the control group. However, a weak reaction in the perinuclear region and along the actin filaments still remained when observed under confocal microscopy. CONCLUSION: These results suggest that the elevation of the expressions of Hsp25 and alphaB crytallin were dependent on the concentration of H2O2. Additionally, the redistribution of these small Hsps is closely associated with the actin filaments, as well as the nuclei. Therefore, much more research will be essential for the elucidation of roles of Hsp25 and alphaB crytallin in oxidative stress, such as ischemia or ischemic-reperfusion injury.
Actin Cytoskeleton ; Actins* ; Blister ; Cell Line* ; Ethanol ; Heart ; Heat-Shock Proteins* ; Hot Temperature* ; Ischemia ; Metals, Heavy ; Microscopy, Confocal ; Microscopy, Electron ; Oxidative Stress ; Reperfusion Injury ; Shock ; Sodium

BACKGROUND: Topoisomerase II (topo II) is a major target of anthracyclines and epipodophyllotoxins for anticancer treatment. The expression of topo II is low in drug resistant cell lines. High levels of glutathione S-transferase (GST)pi have been associated with emergence of cell lines resistant to alkylating agents or adriamycin. METHODS: By immunostaining with paraffin embedded bone marrow tissues, the expression of topo II alpha and GSTpi was investigated in 51 patients with acute myeloid leukemia (AML), and the relation of topo II alpha and GSTpi expression to treatment response in 29 patients with AML following induction chemotherapy was also evaluated. RESULTS: Topo II positive cells varied from less than 1% to 60% of leukemic cells and 20 (39.2%) were negative for topo II (positive cells<10%). Treatment response following chemotherapy was not related to topo II. 26 (51.0%) were positive for GSTpi. GSTpi expression was related to treatment resistance of the patients following chemotherapy. In the patients who showed both topo II alpha negative and GSTpi positive, the frequency of treatment resistance following chemotherapy was high. CONCLUSIONS: This study suggests that immunostaining of topo II alpha and GSTpi with the bone marrow paraffin sections of AML patients can be useful to predict the treatment response following chemotherapy and that further study including more patients with prospective study may substantiate topo II alpha and GSTpi as multidrug resistant markers.
Alkylating Agents ; Anthracyclines ; Bone Marrow ; Cell Line ; DNA Topoisomerases, Type II* ; Doxorubicin ; Drug Resistance, Multiple ; Drug Therapy ; Glutathione S-Transferase pi* ; Glutathione Transferase* ; Glutathione* ; Humans ; Immunohistochemistry ; Induction Chemotherapy ; Leukemia, Myeloid, Acute* ; Paraffin ; Podophyllotoxin

OBJECTIVES: The purpose of this study was to assess physical illnesses, psychological difficulties, and daily life function of victims' family members one year after the Sewol ferry disaster, in order to establish a basis of strategies to promote their mental health and resilience. METHODS: We statistically analyzed self-reported data collected from a survey conducted between March 22 and 28, 2015 with 139 family members of victims. RESULTS: All respondents complained about one or more psychological difficulties, and 95.4% of them had newly diagnosed or exacerbated physical illnesses. An increased number of psychological difficulties was associated with a greater tendency to present physical illnesses including chronic headache, gastrointestinal, musculoskeletal, obstetrico-gynecological, genitourinary, and cardiovascular disease. In addition, the majority of respondents reported a decrease in life functioning. Specifically, 71.9% reported interpersonal avoidance and 63.9% reported difficulty in returning to work. CONCLUSIONS: Not only psychological trauma but also physical illness and daily functioning should be evaluated in detail and actively addressed in the bereaved family members after a catastrophe.
Cardiovascular Diseases ; Disasters ; Headache Disorders ; Humans ; Mental Health ; Psychological Trauma ; Surveys and Questionnaires

Fetomaternal hemorrhage (FMH) is due to the entry of fetal blood into the maternal circulation. Although very rare, FMH complicates pregnancies, presents with severe symptoms, and leads to fetal death. Majority of FMH cases are idiopathic and difficult to diagnose. The known used diagnostic tests are Kleihauer-Betke Test (KBT) and flow cytometry, which can detect fetal hemoglobin in the maternal blood. However, such methods have limited use because of low sensitivity, labor-intensive and error-susceptible procedures, poor reproducibility, and tendency to overestimate the FMH volume. Other tests include high performance liquid chromatography (HPLC) and alpha-fetoprotein (AFP) tests, which can be as favorable to confirm FMH as KBT. However, in case of acute FMH, the diagnostic results of KBT, flow cytometry, and HPLC may be false negative. AFP test is a noninvasive, fast, easily assessable, adjuvant, and confirmatory diagnostic test. Published Korean articles show confirmed FMH by KBT or HPLC in singleton late-preterm and term neonates. Herein, we report a case of monochorionic diamniotic twin neonates (birth weight <1,500 g) who presented borderline fetal hemoglobin level because of acute FMH and were diagnosed with FMH by maternal AFP. Our experience of diagnosing FMH rapidly by AFP test will be very helpful to clinicians for the prevention and treatment of FMH during pregnancy.

Fetomaternal hemorrhage (FMH) is due to the entry of fetal blood into the maternal circulation. Although very rare, FMH complicates pregnancies, presents with severe symptoms, and leads to fetal death. Majority of FMH cases are idiopathic and difficult to diagnose. The known used diagnostic tests are Kleihauer-Betke Test (KBT) and flow cytometry, which can detect fetal hemoglobin in the maternal blood. However, such methods have limited use because of low sensitivity, labor-intensive and error-susceptible procedures, poor reproducibility, and tendency to overestimate the FMH volume. Other tests include high performance liquid chromatography (HPLC) and alpha-fetoprotein (AFP) tests, which can be as favorable to confirm FMH as KBT. However, in case of acute FMH, the diagnostic results of KBT, flow cytometry, and HPLC may be false negative. AFP test is a noninvasive, fast, easily assessable, adjuvant, and confirmatory diagnostic test. Published Korean articles show confirmed FMH by KBT or HPLC in singleton late-preterm and term neonates. Herein, we report a case of monochorionic diamniotic twin neonates (birth weight <1,500 g) who presented borderline fetal hemoglobin level because of acute FMH and were diagnosed with FMH by maternal AFP. Our experience of diagnosing FMH rapidly by AFP test will be very helpful to clinicians for the prevention and treatment of FMH during pregnancy.

Essential oils are mixtures of volatile, lipophilic compounds originating from plants. Some essential oils have useful biological activities including antimicrobial, spasmolytic, antiplasmodial, and insect-repelling activities. In this study, we tested the antimicrobial activity of essential oil prepared from the aromatic plant, Cymbopogon citrates, against three important plant pathogenic and medical microorganisms, Pectobacterium carotovorum, Colletotrichum gloeosporioides, and Aspergillus niger. It effectively inhibited the growth of the bacterium, Pectobacterium carotovorum, in a dose-dependent fashion, and 0.5% of the oil inhibited the growth of bacteria completely. Similarly, the essential oil inhibited the growth of plant pathogenic fungus, Colletotrichum gloeosporioides, and the addition of 1% of essential oil completely inhibited the growth of fungus even after 5 days of culture. Finally, it effectively inhibited the growth of the medically and industrially important fungal species, Aspergillus spp. These results suggest that the essential oil from Cymbopogon citrates may be an environmentally safe alternative to inhibit antimicrobial agents for various uses.
Anti-Infective Agents ; Aspergillus ; Aspergillus niger ; Bacteria ; Citrates ; Colletotrichum ; Cymbopogon ; Fungi ; Oils, Volatile ; Pectobacterium carotovorum ; Plants

PURPOSE: The purpose of this cross-over experimental study was to examine effects of music intervention on maternal anxiety, fetal heart rate pattern and testing time during non-stress tests (NST) for antenatal fetal assessment. METHODS: Sixty pregnant women within 28 to 40 gestational weeks were randomly assigned to either the experimental group (n=30) or control group (n=30). Music intervention was provided to pregnant women in the experimental group during NST. Degree of maternal anxiety and fetal heart rate pattern were our primary outcomes. State-trait anxiety inventory, blood pressure, pulse rate, and changes in peripheral skin temperature were assessed to determine the degree of maternal anxiety. Baseline fetal heart rate, frequency of acceleration in fetal heart rate, fetal movement test and testing time for reactive NST were assessed to measure the fetal heart rate pattern. RESULTS: The experimental group showed significantly lower scores in state anxiety than the control group. There were no significant differences in systolic blood pressure and pulse rate between the two groups. Baseline fetal heart rate was significantly lower in the experimental group than in the control group. Frequency of acceleration in fetal heart rate was significantly increased in the experimental group compared to the control group. There were no significant differences in fetal movement and testing time for reactive NST between the two groups. CONCLUSION: Present results suggest that music intervention could be an effective nursing intervention for alel viating anxiety during non-stress test.
Acceleration ; Anxiety* ; Blood Pressure ; Cardiotocography ; Female ; Fetal Heart* ; Fetal Movement ; Heart Rate ; Heart Rate, Fetal* ; Humans ; Music* ; Nursing ; Pregnancy ; Pregnant Women ; Skin Temperature

BACKGROUND: Fatigue is one of the most common presenting symptoms in primary care in Korea. In this study, we aimed to determine the effect of exercise intervention on the severity of fatigue of unknown medical cause during a period of follow-up. METHODS: We used the data collected from an outpatient fatigue clinic in Seoul National University Bundang Hospital. The study was conducted from March 3, 2010 to May 31, 2014. We measured the body mass index of each patient and evaluated variables including lifestyle factors (smoking, alcohol consumption, and regular exercise), quality of sleep, anxiety, depression, stress severity, and fatigue severity using questionnaires. A total of 152 participants who completed questionnaires to determine changes in fatigue severity and the effect of exercise for each period were evaluated. We used univariate analysis to verify possible factors related to fatigue and then conducted multivariate analysis using these factors and the literature. RESULTS: Of 130 patients with the complaint of chronic fatigue for over 6 months, over 90 percent reported moderate or severe fatigue on the Fatigue Severity Scale and Brief Fatigue Inventory questionnaires. The fatigue severity scores decreased and fatigue improved over time. The amount of exercise was increased in the first month, but decreased afterwards. CONCLUSION: There was no significant relationship between changes in the amount of exercise and fatigue severity in each follow-up period. Randomized controlled trials and a cohort study with a more detailed exercise protocol in an outpatient setting are needed in the future.
Alcohol Drinking ; Anxiety ; Body Mass Index ; Cohort Studies ; Depression ; Fatigue* ; Follow-Up Studies* ; Humans ; Korea ; Life Style ; Metabolic Equivalent ; Motor Activity ; Multivariate Analysis ; Outpatients ; Primary Health Care ; Seoul

Recently, new treatments for human heart disease such as ischemia, infarction, cardiomyopathy, coronary heart disease have been developed. transplantation various kinds of cells from skeletal muscle, endothelium, mesenchyme, hemopoietic tissue to injured area after infarction were challenged. It's so called 'Cell Transplantation'. This therapeutic strategy already adopted and got a good result in clinical trial. But several limitations are still remained, including ethics, donor cell numbers, side effects, therapeutic efficiency. In this research, we investigated the formation of intercellular junction and synchronous contraction between cardiomyocyte and H9c2 cell line in co-culture to establish experimental model in vitro for cell transplantation. For this purpose, two kinds of cells, primary cultured cardiomyocyte and H9c2 (cardiomyoblast cell line) were used. Cultured cardiomyocytes had repetitive contraction-relaxation pattern along longitudinal axis both in single and coculture. But their contractions were slower, less regular, less strong in co-culture than in cardiomyocyte culture only. H9c2 cells did not contracted actively themselves, but moved toward cardiomyocyte passively coincided with contraction. In contact region between two kinds of cells, there was no signal after immunocytochemical staining labeled with connexin43 (gap junction), desmoplakin (desmosome), N-cadherin (adherent junction) even though they had membrane contact. Moreover, F-actin and striation were less developed. These results suggested that co-culture system interfere with remodelling of contractile apparatus, intercellular junction formation as well as contraction-relaxation. Furthermore cardiomyocyte could not induce H9c2 cells differentiation into cardiomyocyte. Therefore, much more research would be essential for clinical application of cell transplantation and this study would be the basic source for further study of new therapy of myocardial disease and building up in vitro model.
Actins ; Axis, Cervical Vertebra ; Cadherins ; Cardiomyopathies ; Cell Count ; Cell Line* ; Cell Transplantation ; Coculture Techniques* ; Connexin 43 ; Coronary Disease ; Desmoplakins ; Endothelium ; Ethics ; Heart Diseases ; Humans ; Infarction ; Intercellular Junctions* ; Ischemia ; Membranes ; Mesoderm ; Models, Theoretical ; Muscle, Skeletal ; Myocytes, Cardiac* ; Tissue Donors ; Transplants