Studies on coat protein I (COPI) have contributed to a basic understanding of how coat proteins generate vesicles to initiate intracellular transport. The core component of the COPI complex is coatomer, which is a multimeric complex that needs to be recruited from the cytosol to membrane in order to function in membrane bending and cargo sorting. Previous structural studies on the clathrin adaptors have found that membrane recruitment induces a large conformational change in promoting their role in cargo sorting. Here, pursuing negative-stain electron microscopy coupled with single-particle analyses, and also performing CXMS (chemical cross-linking coupled with mass spectrometry) for validation, we have reconstructed the structure of coatomer in its soluble form. When compared to the previously elucidated structure of coatomer in its membrane-bound form we do not observe a large conformational change. Thus, the result uncovers a key difference between how COPI versus clathrin coats are regulated by membrane recruitment.
ADP-Ribosylation Factor 1 ; chemistry ; metabolism ; Animals ; Coatomer Protein ; chemistry ; metabolism ; Cytosol ; chemistry ; metabolism ; GTPase-Activating Proteins ; chemistry ; metabolism ; Humans ; Membranes, Artificial ; Rats

We have previously isolated epsilon-COP, the alpha-COP interactor in COPI of Aspergillus nidulans, by yeast two-hybrid screening. To understand the function of epsilon-COP, the aneA+ gene for epsilon-COP/AneA was deleted by homologous recombination using a gene-specific disruption cassette. Deletion of the epsilon-COP gene showed no detectable changes in vegetative growth or asexual development, but resulted in decrease in the production of the fruiting body, cleistothecium, under conditions favorable for sexual development. Unlike in the budding yeast Saccharomyces cerevisiae, in A. nidulans, over-expression of epsilon-COP did not rescue the thermo-sensitive growth defect of the alpha-COP mutant at 42degrees C. Together, these data show that epsilon-COP is not essential for viability, but it plays a role in fruiting body formation in A. nidulans.
Aspergillus nidulans* ; Coatomer Protein* ; Fruit ; Homologous Recombination ; Mass Screening ; Saccharomyces cerevisiae ; Saccharomycetales ; Sexual Development ; Yeasts

The aim of this study was to applicate and evaluate a SYBR Green real-time PCR for the specific detection of Salmonella spp. Specificity of the PCR method was confirmed with 48 Salmonella spp. and 5 non-Salmonella strains using invA gene primer. The average threshold cycle (C(T)) of Salmonella spp. was 11.83 +/- 0.78 while non-Salmonella spp. was 30.86 +/- 1.19. Correlation coefficients of standard curves constructed using C(T) versus copy number of Salmonella Enteritidis ATCC 13076 showed good linearity (R2 = 0.993; slope = 3.563). Minimum level of detection with the method was > 10(2) colony forming units (CFU)/mL. These results suggested that the SYBR Green real-time PCR might be applicable for the specific detection of Salmonella spp. isolates.
Coat Protein Complex I ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Salmonella ; Salmonella enteritidis ; Sensitivity and Specificity ; Stem Cells

OBJECTIVE: To investigate the effect of brain-derived neurotrophic factor (BDNF) Val66Met polymorphism on the recovery after subcortical stroke, using the modified Rankin Scale (mRS). METHODS: Subcortical stroke patients with copies of BDNF Val66Met polymorphism (n=7) were compared to their controls (n=7) without a copy of BDNF Val66Met polymorphism after matching for initial severity, location and type of stroke. The mRS scores at 1 and 3 months after discharge from the neurorehabilitation unit were compared between the groups. RESULTS: A repeated measures ANOVA for mRS revealed significant interaction between time and group (F(2, 24) =37.2, p<0.001) and a significant effect of time (F(2, 24)=10.8, p<0.001), thereby reflecting significant differences between the Met allele (+) group and the Met allele (-) group. There was a significant difference in mRS scores at 3 months post-discharge between the two groups (p=0.01) although no difference was evident in mRS scores at 1 month post-discharge between the two groups. There were significant improvements between mRS scores on admission and mRS scores at 1 month post-discharge (p=0.02), and between mRS scores at 1 month post-discharge and mRS scores at 3 months post-discharge (p=0.004) in the Met allele (-) group. CONCLUSION: BDNF Val66Met polymorphism may be associated with worse functional outcome in Korean patients with subcortical stroke. Therefore, BDNF Val66Met polymorphism should be considered as an important prognostic factor for recovery and responses to rehabilitation therapies after stroke in Korean patients. There is a need for developing different rehabilitation strategies for the population with BDNF Val66Met polymorphism. Further studies assessing different outcomes for various functional domains of stroke recovery are needed to clarify the role of BDNF Val66Met polymorphism.
Alleles ; Brain-Derived Neurotrophic Factor ; Coat Protein Complex I ; Humans ; Stroke

BACKGROUND/AIMS: It is suggested that the hepatic lipid composition is more important than lipid quantity in the pathogenesis of non-alcoholic steatohepatitis. We examined whether lipoic acid (LA) could alter intrahepatic lipid composition and free cholesterol distribution. METHODS: HepG2 cells were cultured with palmitic acid (PA) with and without LA. Apoptosis, changes of the mitochondrial structure, intracellular lipid partitioning, and reactive oxygen species (ROS) activity were measured. RESULTS: Free fatty acid (FA) increased apoptosis, and LA co-treatment prevented this lipotoxicity (apoptosis in controls vs PA vs PA+LA, 0.5% vs 19.5% vs 1.6%, p<0.05). LA also restored the intracellular mitochondrial DNA copy number (553+/-33.8 copies vs 291+/-14.55 copies vs 421+/-21.05 copies, p<0.05) and reversed the morphological changes induced by PA. In addition, ROS was increased in response to PA and was decreased in response to LA co-treatment (41,382 relative fluorescence unit [RFU] vs 43,646 RFU vs 41,935 RFU, p<0.05). LA co-treatment increased the monounsaturated and polyunsaturated FA concentrations and decreased the total saturated FA fraction. It also prevented the movement of intracellular free cholesterol from the cell membrane to the cytoplasm. CONCLUSIONS: LA opposes free FA-generated lipotoxicity by altering the intracellular lipid composition and free cholesterol distribution.
Apoptosis ; Cell Membrane ; Cholesterol ; Coat Protein Complex I ; DNA, Mitochondrial ; Fatty Liver ; Fluorescence ; Hep G2 Cells ; Liver Cirrhosis ; Palmitic Acid ; Reactive Oxygen Species ; Thioctic Acid

Tetrasomy 18p, one of the most commonly observed isochromosomes, consists of two copies of the p arms on chromosome 18[i(18p)]. It is known as a de novo occurrence of non-disjunction or centromeric mis-division during meiosis II in the vast majority of cases. It has a prevalence of 1/140,000-180,000 live births and affects both genders equally. A 28-year-old woman was referred at 33+2 weeks gestation to rule out fetal congenital heart disease. Her prenatal ultrasonography showed intrauterine growth retardation, cardiomegaly, and imperforate anus. Doppler ultrasonographic finding showed fetal anemia. Tetrasomy 18p was confirmed by conventional karyotyping and fluorescence in situ hybridization. Because of its very low prevalence rate, only several cases of tetrasomy 18p has been reported worldwide and it has not yet been reported in Korea before. Therefore, we report a case of prenatally diagnosed tetrasomy 18p.
Anemia ; Aneuploidy ; Anus, Imperforate ; Arm ; Cardiomegaly ; Chromosomes, Human, Pair 18 ; Coat Protein Complex I ; Female ; Fetal Growth Retardation ; Fluorescence ; Heart Diseases ; Humans ; In Situ Hybridization ; Isochromosomes ; Karyotyping ; Korea ; Live Birth ; Meiosis ; Pregnancy ; Prenatal Diagnosis ; Prevalence ; Tetrasomy ; Ultrasonography, Prenatal

PURPOSE: This study was conducted to identify awareness levels and influencing factors of sexual harassment and gender egalitarianism among college students. METHODS: This study was based on a questionnaire survey with a total of 1,244 college students. The copies of the questionnaire were collected from November 2 to December 20, 2011. RESULTS: The average level of sexual harassment and gender egalitarianism among students from the Youngnam province was higher than those of other province. The average level of the sexual harassment and gender egalitarianism among those students who were majoring in healthcare sciences was higher than that among those who were majoring in other subjects than arts and sports sciences. The influencing factors on the recognition of gender egalitarianism were genders and sexual experiences. The influencing factors on the recognition of sexual harassment were genders and ages. CONCLUSION: This study identified college students' awareness of sexual harassment, gender egalitarianism and influencing factors associated with them. It is expected that this study will be helpful for developing strategies for the prevention of sexual harassment for college students.
Coat Protein Complex I ; Delivery of Health Care ; Humans ; Sexual Harassment ; Sports ; Surveys and Questionnaires

PURPOSE: The prescription drug list for primary treatment by community health practitioners has been maintained for 30 years without any modification. Thus, this study will suggest an improvement scheme of prescription drug list for primary health care posts through an analysis of drug use in those posts. METHODS: A questionnaire survey was implemented with community health practitioners from April to June in 2012. A total of 1,249 copies were analyzed. As for the databases of drug use in the integrated information, a total of 154,229 diagnoses selected in the method of stratified cluster sampling from 39 primary health care posts'data were analyzed. We consulted some experts about the prescription medication list, and referred to the medication information on-line home page for up-to-date drug information. RESULTS: This study ultimately suggests 77 prescription drug items for primary health care posts by eliminating 35 items and replacing 1 item from the original list, and adding 4 items to it. CONCLUSION: This study will provide basic data for revising the prescription drug list in primary health care posts by periodically reflecting adverse effects in the existing drugs, demographic and environmental changes, and development of new drugs.
Coat Protein Complex I ; Prescriptions ; Primary Health Care ; Rural Health ; Surveys and Questionnaires

Although physiological function of alpha-synuclein is not yet clearly understood, accumulating evidence strongly suggests it plays a crucial role in the pathogenesis of Parkinson disease. Pathologically, alpha-synuclein is a major component of Lewy bodies, which is the pathological hallmark of Parkinson disease. Alpha-synuclein pathology is observed in the brainstem nuclei, including the dorsal motor nucleus of the vagus nerve, the locus ceruleus, and the substantia nigra in the early phase of Parkinson disease and it may 'spread' to cerebral cortical areas in the advanced Parkinson disease and appears to have a role in the cognitive decline in Parkinson disease. Recently, it is suggested that alpha-synuclein pathology in Parkinson disease starts in the olfactory bulb or enteric nervous system and then spreads to the brainstem. In accordance with this hypothesis, alpha-synuclein pathology has been found in gastric mucosa and colonic mucosa of patients with Parkinson disease. Genetically, SNCA mutations including point mutation and copy number variation are known to cause familial Parkinson disease, further supporting the assumption that alpha-synuclein plays a crucial role in Parkinson disease pathogenesis. In addition, recent GWAS studies consistently show that the SNPs in SNCA genes are associated with risk for sporadic Parkinson disease. It is also known that variations in the promoter region or 3'UTR of SNCA, which increases the expression of SNCA, are associated with the risk for Parkinson disease. Collectively, these findings suggest that further studies on alpha-synuclein will lead to the elucidation of the mechanism of and therapy for Parkinson disease.
3' Untranslated Regions ; alpha-Synuclein ; Brain Stem ; Coat Protein Complex I ; Colon ; Enteric Nervous System ; Gastric Mucosa ; Humans ; Lewy Bodies ; Locus Coeruleus ; Mucous Membrane ; Olfactory Bulb ; Parkinson Disease ; Point Mutation ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic ; Substantia Nigra ; Vagus Nerve

This clinical report presents the reconstruction of a maxillary arch with a cement retained implant supported fixed prosthesis using a monolithic zirconia generated by CAD/CAM system on eight osseointegrated implants. The prosthesis was copy milled from an interim prosthesis minimizing occlusal adjustments on the definitive prosthesis at the time of delivery. Monolithic zirconia provides high esthetics and reduces the number of metal alloys used in the oral cavity.
Alloys ; Coat Protein Complex I ; Esthetics ; Mouth ; Occlusal Adjustment ; Prostheses and Implants ; Zirconium