A beta-glucan synthase gene was isolated from the genomic DNA of polypore mushroom Sparassis crispa, which reportedly produces unusually high amount of soluble beta-1,3-glucan (beta-glucan). Sequencing and subsequent open reading frame analysis of the isolated gene revealed that the gene (5,502 bp) consisted of 10 exons separated by nine introns. The predicted mRNA encoded a beta-glucan synthase protein, consisting of 1,576 amino acid residues. Comparison of the predicted protein sequence with multiple fungal beta-glucan synthases estimated that the isolated gene contained a complete N-terminus but was lacking approximately 70 amino acid residues in the C-terminus. Fungal beta-glucan synthases are integral membrane proteins, containing the two catalytic and two transmembrane domains. The lacking C-terminal part of S. crispa beta-glucan synthase was estimated to include catalytically insignificant transmembrane alpha-helices and loops. Sequence analysis of 101 fungal beta-glucan synthases, obtained from public databases, revealed that the beta-glucan synthases with various fungal origins were categorized into corresponding fungal groups in the classification system. Interestingly, mushrooms belonging to the class Agaricomycetes were found to contain two distinct types (Type I and II) of beta-glucan synthases with the type-specific sequence signatures in the loop regions. S. crispa beta-glucan synthase in this study belonged to Type II family, meaning Type I beta-glucan synthase is expected to be discovered in S. crispa. The high productivity of soluble beta-glucan was not explained but detailed biochemical studies on the catalytic loop domain in the S. crispa beta-glucan synthase will provide better explanations.
Agaricales ; Cell Wall ; Classification ; Clone Cells* ; Cloning, Organism* ; DNA ; Efficiency ; Exons ; Glycogen Synthase ; Humans ; Introns ; Membrane Proteins ; Open Reading Frames ; RNA, Messenger ; Sequence Analysis

A chemical mutagenesis technique was employed for development of mutant strains of Sparassis crispa targeting the shortened cultivation time and the high beta-glucan content. The homogenized mycelial fragments of S. crispa IUM4010 strain were treated with 0.2 vol% methyl methanesulfonate, an alkylating agent, yielding 199 mutant strains. Subsequent screening in terms of growth and beta-glucan content yielded two mutant strains, B4 and S7. Both mutants exhibited a significant increase in beta-glucan productivity by producing 0.254 and 0.236 mg soluble beta-glucan/mg dry cell weight for the B4 and S7 strains, respectively, whereas the wild type strain produced 0.102 mg soluble beta-glucan/mg dry cell weight. The results demonstrate the usefulness of chemical mutagenesis for generation of mutant mushroom strains.
Agaricales ; Efficiency ; Mass Screening ; Mesylates ; Methyl Methanesulfonate ; Mutagenesis ; Sprains and Strains

There are four principles of medical ethics; Beneficence, Respect for autonomy, Non-maleficence, and Justice. It is not easy to apply to principles of medical ethics in the special circumstances of obstetrics and gynecology. Student doctors must learn to be familiar with principles of medical ethics tailored to the special circumstances while the obstetrics and gynecology practice.
Beneficence ; Education, Medical* ; Ethics* ; Ethics, Medical ; Gynecology* ; Humans ; Obstetrics* ; Social Justice

PURPOSE: This study was undertaken to observe the pattern of methylation of the p16INK4a and human telomerase reverse transcriptase (hTERT) genes and the p16 and hTERT protein expressions in invasive ductal carcinoma of the breast. In addition, we evaluated the relationship between the methylation status of the two genes and their protein expressions. METHODS: We performed methylation-specific PCR (MSP) and immunohistochemical staining in 63 breast cancer specimens. RESULTS: There was no statistical association between p16INK4a gene methylation and the histological grade (tumor grade, tumor size and lymph node status). Methylation of the hTERT promoter did show significant differences according to the histological tumor grade and tumor size, but there was no clinical significance. Methylation of the p16INK4a and hTERT genes was found in 22.2% and 31.8% of the specimens, respectively. A negative p16 protein expression (0-10% expression rate) was observed in 38.1% of the specimens (24 of 63). A positive hTERT expression (more than a 25% expression rate) was observed in 73.0% of the specimens (46 of 63). There was no statistical significance in the relationship between the methylation status and the protein expression. CONCLUSION: Our data suggest that methylation of the p16 and hTERT genes is not associated with their protein expressions according to Immunohistochemisty. There seemed to be another complicated mechanism for p16 inactivation and hTERT activation in breast cancer.
Breast Neoplasms* ; Breast* ; Carcinoma, Ductal ; Genes, p16 ; Humans* ; Lymph Nodes ; Methylation ; Polymerase Chain Reaction ; Telomerase

Chemical mutagenesis of basidiospores of Hypsizygus marmoreus generated new mushroom strains. The basidospores were treated with methanesulfonate methylester, an alkylating agent, to yield 400 mutant monokaryotic mycelia. Twenty fast-growing mycelia were selected and mated each other by hyphal fusion. Fifty out of the 190 matings were successful (mating rate of 26.3%), judged by the formation of clamp connections. The mutant dikaryons were cultivated to investigate their morphological and cultivation characteristics. Mutant strains No. 3 and No. 5 showed 10% and 6% increase in fruiting body production, respectively. Eight mutant strains showed delayed and reduced primordia formation, resulting in the reduced production yield with prolonged cultivation period. The number of the fruiting bodies of mutant No. 31, which displayed reduced primordial formation, was only 15, compared to the parental number of 65. Another interesting phenotype was a fruiting body with a flattened stipe and pileus. Dikaryons generated by mating with the mutant spore No. 14 produced flat fruiting bodies. Further molecular biological studies will provide details of the mechanism. This work shows that the chemical mutagenesis approach is highly utilizable in the development of mushroom strains as well as in the generation of resources for molecular genetic studies.
Agaricales ; Breeding ; Fruit ; Humans ; Mesylates ; Molecular Biology ; Mutagenesis ; Parents ; Phenotype ; Spores

PURPOSE: inhaled nitric oxide(iNO) is an excellent method for the postoperative pulmonary hypertension in congenital heart disease. But more detailed care is needed because of the development of rebound pulmonary hypertension after NO Withdrawal. We performed this study in order to discontinue the iNO successfully by way of presenting the adequate weaning and supplying methods. METHODS: Between January, 1998 and August, 1999 we sudied 10 patients who had rebound pulmonary hypertension(RPH) after iNO withdrawal. We completed the iNO in these patween the first the second trial of the weaning process. We tried to discover the differences between the first and second weaning process. We measured NO concentration at the start and just before NO withdrawal and during the period of weaning process. Moreover, to identify the iNO effects during the weaning of the iNO, we counted the degree of the change of PaO2/FiO2and mean PAP/SAP beween initial and at half of the initial NO concentration. RESULTS: Second weaning had a longer duration weaning process(11+/-0 cersus 5+/- hours, P<0.05), lower NO concentration just before NO withdrawal(2+/-.6 versus 4+/-ppm, P<0.05). In the change of the mean PAP/SAP and PaO2/FiO2as iNO was weaning from the initial iNO concentration to a half of the initial iNO concentration, the degree of increase in mean PAP/SAP(0.026+/-.07 versus 0.054+/-.07, P<0.05) and the degree of decrease in PaO2/FiO2(49+/-4 versus 65+/-2, P<0.05) were smaller in the second in the second weaning process than the first weaning process. CONCLUSION: A successful weaning of iNO can be performed with a low iNO concentration at the start and just before withdrawal and with the long duration iNO weaning process. Moreover, We speculate that the degree of change in the mean PAP/SAP and PaO2/FiO2at the half of the iNO weaning process are an indicator for the development of RPH.
Heart Defects, Congenital ; Humans ; Hypertension, Pulmonary ; Nitric Oxide* ; Weaning*

PURPOSE: The effect of PTK inhibitors (herbimycin A and genistein) on the induction of radiation-induce d apoptosis in Ph-positive K562 leukemia cell line was investigated. MATERIALS AND METHODS: K562 cells in exponential growth phase were irradiated with a linear accelerator at room temperature. For 6 MV X-ray irradiation and drug treatment, cultures were initiated at 2x10' cells/mL. The cells were irradiated with 10 Gy. Stock solutions of herbimycin A and genistein were prepared in dimethyl sulphoxide (DMSO). After incubation at 37C for 0-48 h, the extent of apoptosis was determined using agarose gel electrophoresis and TUNEL assay. The progression of cells throughth the cel l cycle after irradiation and drug treatment was also determined with flow cytometry. Western blot analysis was used to monitor bcl-2, bcl-X and bax protein levels. RESULTS: Treatment with 10 Gy X-irradiation did not result in the induction of apoptosis. The HMA alone (500 nM) also failed to induce apoptosis. By contrast, incubation of K562 cells with HMA after irradiation resulted in a substantial induction of nuclear condensation and fragmentation by agarose gel electro-phoresis and TUNEL assay. Genistein failed to enhance the ability of X-irradiation to induce DN A fragmentation. Enhancement of apoptosis by H MA was not attributable to downregulation of the bcl-2 or bcl-X anti-apoptotic proteins. When the cells were irradiated and maintained with HMA, the percentage cf cells in G2/M phase decreased to 30-40% at 48 h. On the other hand, cells exposed to 10 Gy X-irradiation alone or maintained with genistein did not show marked cell cycle redistribution. CONCLUSION: We have shown that nanomolar concentrations of the PTK inhibitor HMA synergize with X-irradiation in inducing the apoptosis in Ph (+) K562 leukemia cell line. While, genistein, a PTK inhibitor which is not selective for p2 10""'' failed to enhance the radiation induced apoptosis in K562 cells. It is unlikely that the ability of HMA to enhance apoptosis in K562 cells is attributable to bcl-2 family. It is plausible that the relationship between cell cycle delays and cell death is essential for drug development based on molecular targeting designed to modify radiation-induced apoptosis.
Apoptosis Regulatory Proteins ; Apoptosis* ; bcl-2-Associated X Protein ; Blotting, Western ; Cell Cycle ; Cell Death ; Cell Line ; Dimethyl Sulfoxide ; Down-Regulation ; Electrophoresis, Agar Gel ; Flow Cytometry ; Genistein ; Hand ; Humans ; Hydrogen-Ion Concentration ; In Situ Nick-End Labeling ; K562 Cells* ; Leukemia ; Particle Accelerators ; Sepharose

Fractures, Comminuted ; Humans ; Incidence ; Mandible ; Mandibular Fractures ; Osteomyelitis ; Postoperative Complications ; Retrospective Studies ; Treatment Outcome

BACKGROUND: It is well known that patients with neurological disorders are vulnerable to depression. However, in Korea, National Health Insurance services had banned non-psychiatrists from prescribing antidepressants for more than 2 months until January 2017. Now, neurologists are able to prescribe antidepressants to patients with only four neurological disorders. Due to this recent change in national health insurance policy, there will be a large change in the prescription pattern of antidepressants. In this study, we performed an analysis of antidepressant prescription patterns in Korea prior to this recent policy change. METHODS: The source population of this retrospective cohort study is the Health Insurance Review & Assessment Service database. We analyzed the claim database for patients who have one of four major neurologic disorders and had healthcare documentation submitted by healthcare providers between January 1, 2011 and December 31, 2016. RESULTS: During 2012–2016, antidepressant prescription rates of 6.21% (127,192 of a total 2,048,165 patients), 9.93% (81,861 out of 824,290), 10.12% (173,582 of 1,714,776), and 13.36% (48,530 of 363,347) were found for cerebrovascular disease, epilepsy, dementia, and Parkinson's disease respectively. The most frequently prescribed antidepressant in cerebrovascular disease and epilepsy was tricyclic antidepressants (TCAs). In Parkinson's disease and dementia, the most frequently used antidepressant was selective serotonin reuptake inhibitors. CONCLUSIONS: The overall prescription rate of antidepressants was much lower than the estimated rates reported in other countries. TCAs were the primarily prescribed antidepressant. It is now expected that TCAs will be replaced by newer antidepressants.
Antidepressive Agents ; Antidepressive Agents, Tricyclic ; Cerebrovascular Disorders ; Cohort Studies ; Delivery of Health Care ; Dementia ; Depression ; Epilepsy ; Health Personnel ; Humans ; Insurance, Health ; Korea ; National Health Programs ; Nervous System Diseases ; Parkinson Disease ; Prescriptions ; Retrospective Studies ; Serotonin Uptake Inhibitors