41 results on '"Kohji Murakami"'
Search Results
2. Relationship between mortality and Council of Nutrition Appetite Questionnaire scores in Japanese nursing home residents
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Yutaka Watanabe, Jun Yasuda, Kohji Murakami, Junichi Furuya, Yurie Mikami, Hirohiko Hirano, Yu Taniguchi, Maki Shirobe, Keiko Motokawa, Masaharu Murakami, and Ayako Edahiro
- Subjects
Male ,0301 basic medicine ,Gerontology ,Clinical Dementia Rating ,Barthel index ,Endocrinology, Diabetes and Metabolism ,media_common.quotation_subject ,Appetite ,Nutritional Status ,030209 endocrinology & metabolism ,Cohort Studies ,03 medical and health sciences ,0302 clinical medicine ,Japan ,Surveys and Questionnaires ,Homes for the Aged ,Humans ,Medicine ,Medical history ,Mortality ,Geriatric Assessment ,media_common ,Aged, 80 and over ,030109 nutrition & dietetics ,Nutrition and Dietetics ,business.industry ,Mean age ,Nursing Homes ,Nutrition Assessment ,Mortality data ,Female ,business ,Nursing homes ,Cohort study - Abstract
Objective This 1-y cohort study examined whether Council of Nutrition Appetite Questionnaire (CNAQ) scores predicted mortality in 316 elderly Japanese residents of five nursing homes (60 men, 256 women; mean age: 84.9 ± 8.3 y). Methods The baseline survey included participant characteristics (e.g., age, sex, height, weight, and medical history), and Barthel Index (BI), Clinical Dementia Rating (CDR), Mini Nutritional Assessment-Short Form (MNA®-SF), CNAQ, Simplified Nutritional Appetite Questionnaire (SNAQ; simplified CNAQ), and SNAQ for the Japanese elderly (SNAQ-JE) scores. Results Following the baseline survey, mortality data were collected for 1 y; during this time, 62 participants (19.6%) died. The deceased group's CNAQ scores (25.1 ± 4.8) were significantly lower than those of the survival group (28 ± 3.6; P Conclusions This study showed that CNAQ scores were inversely associated with 1-y mortality. Furthermore, appetite assessment using the CNAQ predicted the death of Japanese nursing home residents. Similarly, the SNAQ and SNAQ-JE scores were inversely associated with 1-y mortality.
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- 2019
3. Effects of lactoferrin and lactoperoxidase-containing food on the oral microbiota of older individuals
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Fumiaki Abe, Kohji Murakami, Shouji Hironaka, Manabu Nakano, Kentaro Ishikawa, Toshitaka Odamaki, Hiroyuki Wakabayashi, Jin-zhong Xiao, Hirosuke Sugahara, and Koji Yamauchi
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0301 basic medicine ,Saliva ,Lactoferrin ,Immunology ,Lactoperoxidase ,Physiology ,030206 dentistry ,Biology ,Dental plaque ,medicine.disease ,Microbiology ,stomatognathic diseases ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,medicine.anatomical_structure ,Tongue ,Virology ,Oral microbiology ,biology.protein ,medicine ,Ingestion ,Anaerobic bacteria - Abstract
The oral microbiota influences health and disease states. Some gram-negative anaerobic bacteria play important roles in tissue destruction associated with periodontal disease. Lactoferrin (LF) and lactoperoxidase (LPO) are antimicrobial proteins found in saliva; however, their influence on the whole oral microbiota currently remains unknown. In this randomized, double-blinded, placebo-controlled study, the effects of long-term ingestion of LF and LPO-containing tablets on the microbiota of supragingival plaque and tongue coating were assessed. Forty-six older individuals ingested placebo or test tablets after every meal for 8 weeks. The relative abundance of bacterial species was assessed by 16S rRNA gene high-throughput sequencing. Most of the bacterial species in supragingival plaque and tongue coating that exhibited significant decreases in the test group were gram-negative bacteria, including periodontal pathogens. Decreases in the total relative abundance of gram-negative organisms in supragingival plaque and tongue coating correlated with improvements in assessed variables related to oral health, such as oral malodor and plaque accumulation. Furthermore, there was significantly less microbiota diversity in supragingival plaque at 8 weeks in the test group than in the placebo group and low microbiota diversity correlated with improvements in assessed variables related to oral health. These results suggest that LF and LPO-containing tablets promote a shift from a highly diverse and gram-negative-dominated to a gram-positive-dominated community in the microbiota of supragingival plaque and tongue coating. This microbial shift may contribute to improvements in oral health, including oral malodor and state of the gingiva.
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- 2017
4. Factors affecting masticatory function of community‐dwelling older people: Investigation of the differences in the relevant factors for subjective and objective assessment
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Yuki Ohara, Yutaka Watanabe, Kohji Murakami, Shoji Shinkai, Shouji Hironaka, Masaharu Murakami, Akihiko Kitamura, Daisuke Takagi, Yu Taniguchi, Hirohiko Hirano, and Ayako Edahiro
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Male ,medicine.medical_specialty ,Oral Health ,Bite Force ,Objective assessment ,03 medical and health sciences ,Health examination ,Cognition ,0302 clinical medicine ,030502 gerontology ,Activities of Daily Living ,Humans ,Medicine ,Gait ,General Dentistry ,Aged ,Hand Strength ,Depression ,business.industry ,030206 dentistry ,Odds ratio ,Confidence interval ,Gait speed ,Masticatory force ,Physical therapy ,Mastication ,Female ,Independent Living ,Geriatrics and Gerontology ,0305 other medical science ,business ,Older people - Abstract
Objective This study aimed to examine relevant factors for subjective and objective assessment of masticatory functions and elucidate any differences between the two methods. Background Previous studies have reported that the results of subjective and objective assessment of masticatory function in older people do not necessarily agree. Materials and methods This study included 487 community-dwelling Japanese older people (205 male and 282 female; mean age 74.1±6.3 years) who participated in a comprehensive geriatric health examination. Basic information (gender and age), higher level of competence in daily living, depression, subjective masticatory function (SMF) and objective masticatory function (OMF) assessments, cognitive function, skeletal muscle mass, handgrip strength, gait speed and oral status (number of remaining and functional teeth, mouth dryness and occlusal force) were recorded. Results Multiple logistic regression analysis showed that depression (odds ratio [OR]: 1.181, 95% confidence interval [CI]: 1.094-1.275), mouth dryness (OR: 2.037, CI: 1.212-3.423) and occlusal force (OR: 0.997, CI: 0.996-0.999) were significantly associated with SMF, whereas higher level of competence in daily living (OR: 0.730 CI: 0.586-0.910), skeletal muscle mass (OR: 0.521 CI: 0.283-0.960), number of functional teeth (OR: 0.862 CI: 0.775-0.959), number of remaining teeth (OR: 0.868 CI: 0.810-0.930) and occlusal force (OR: 0.994, CI: 0.991-0.998) were associated with OMF. Conclusion Subjective masticatory functionSMF and OMF were associated with different factors, suggesting that both mental and physical factors should be taken into consideration when treating decreased masticatory function.
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- 2017
5. Examination of factors affecting the intraoral perception of object size: a preliminary study
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Takafumi Ooka, Shouji Hironaka, Kaori Tomita, Mari Takahashi, and Kohji Murakami
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Adult ,Male ,medicine.medical_specialty ,genetic structures ,media_common.quotation_subject ,Illusion ,Object (grammar) ,Oropharynx ,Dentistry ,Audiology ,Oral cavity ,Young Adult ,03 medical and health sciences ,0302 clinical medicine ,Perception ,Sensory threshold ,Humans ,Medicine ,Particle Size ,Set (psychology) ,General Dentistry ,Size Perception ,media_common ,Mouth ,business.industry ,030206 dentistry ,Healthy Volunteers ,Deglutition ,Sensory function ,Food ,Sensory Thresholds ,030220 oncology & carcinogenesis ,Mastication ,Female ,business - Abstract
Oral sensory function is essential for the successful performance of a range of ingestion. Although the perception of object size is important in determining the degree of manipulation, evidence suggests that people does not always perceive the size of the object in oral cavity accurately. The purpose of this study was to investigate the factors affecting the intraoral perception of object size. Twenty-three healthy young volunteers detected the size of cylinders inserted into oral cavity blindly and identified it from the reference set. The diameter of cross section varied from 10 to 14 mm in a gap of 1 mm and had three different temperatures (5, 36, 50 °C) in each. The perceived size was recorded, and the difference with the actual size was assessed. The required time for identification was also measured and compared between gender and between temperatures. The results demonstrated that the direction of size illusion was significantly affected by the required time for identification. Long manipulation led to overestimation, and short manipulation led to underestimation of object size irrespective of temperature and size. Gender was the other factor affecting intraoral size perception. The rate of overestimation was low in female participants comparing with male participants in this experiment, although the number of participants was limited. We therefore concluded that in order to detect the other factors affecting intraoral size perception, regulating oral manipulation time is indispensable.
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- 2017
6. Relationship between skeletal muscle mass and swallowing function in patients with Alzheimer's disease
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Hunkyung Kim, Yutaka Watanabe, Kohji Murakami, Daisuke Takagi, Hirohiko Hirano, Shouji Hironaka, Ayako Edahiro, Hideyo Yoshida, and Yuki Ohara
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medicine.medical_specialty ,business.industry ,Clinical Dementia Rating ,Odds ratio ,medicine.disease ,Confidence interval ,03 medical and health sciences ,0302 clinical medicine ,Swallowing ,030502 gerontology ,Internal medicine ,Sarcopenia ,Physical therapy ,Medicine ,Dementia ,030212 general & internal medicine ,Alzheimer's disease ,0305 other medical science ,business ,Body mass index - Abstract
Aim The present study verified the hypothesis that decreased skeletal muscle in older adults with Alzheimer's disease is related to Alzheimer's disease progression and decreased oral or swallowing function. Methods We investigated 232 patients with Alzheimer's disease (31 men, 201 women, average age 85.4 ± 5.9 years) in two regions in Japan. The patients provided basic information (sex and age), and were assessed for skeletal muscle index, dementia severity (clinical dementia rating), activities of daily living, nutritional status, oral status and swallowing function. Results Stratification by clinical dementia rating was as follows: clinical dementia rating 0.5:21 patients (9.0%), clinical dementia rating 1:85 patients (36.6%), clinical dementia rating 2:88 patients (37.9%) and clinical dementia rating 3:38 patients (16.3%). Alzheimer's disease severity was significantly related to skeletal muscle index. Logistic regression analysis showed that clinical dementia rating 3 (odds ratio 11.68, 95% confidence interval 4.52–30.20), body mass index
- Published
- 2016
7. Clinical Statistics for Dysphagia Patients ≤18 Years of Age in the Center of Special Needs Dentistry, April 2012–March 2013
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Kazumi Kubota, Yoshiharu Mukai, Akemi Utsumi, Toshikazu Ishikawa, Keigo Ishida, Takuya Asami, Kohji Murakami, Masahiro Watanabe, Asari Ebuchi, Luna Osakabe, Kaori Tomita, Mari Takahashi, Daisuke Takagi, Kentaro Ishikawa, Shouji Hironaka, Yu Morita, Takafumi Ooka, Akiko Ishizaki, and Ayano Ogawa
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Clinical statistics ,medicine.medical_specialty ,Pathology ,business.industry ,General surgery ,medicine ,Special needs ,medicine.symptom ,business ,Dysphagia - Published
- 2015
8. Effects of lactoferrin and lactoperoxidase-containing food on the oral microbiota of older individuals
- Author
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Manabu, Nakano, Hiroyuki, Wakabayashi, Hirosuke, Sugahara, Toshitaka, Odamaki, Koji, Yamauchi, Fumiaki, Abe, Jin-Zhong, Xiao, Kohji, Murakami, Kentaro, Ishikawa, and Shouji, Hironaka
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Aged, 80 and over ,DNA, Bacterial ,Male ,Bacteria ,Microbiota ,Microbial Consortia ,Dental Plaque ,Gingiva ,Oral Health ,Biodiversity ,Lactoferrin ,Double-Blind Method ,Tongue ,RNA, Ribosomal, 16S ,Humans ,Female ,Lactoperoxidase ,Saliva ,Aged - Abstract
The oral microbiota influences health and disease states. Some gram-negative anaerobic bacteria play important roles in tissue destruction associated with periodontal disease. Lactoferrin (LF) and lactoperoxidase (LPO) are antimicrobial proteins found in saliva; however, their influence on the whole oral microbiota currently remains unknown. In this randomized, double-blinded, placebo-controlled study, the effects of long-term ingestion of LF and LPO-containing tablets on the microbiota of supragingival plaque and tongue coating were assessed. Forty-six older individuals ingested placebo or test tablets after every meal for 8 weeks. The relative abundance of bacterial species was assessed by 16S rRNA gene high-throughput sequencing. Most of the bacterial species in supragingival plaque and tongue coating that exhibited significant decreases in the test group were gram-negative bacteria, including periodontal pathogens. Decreases in the total relative abundance of gram-negative organisms in supragingival plaque and tongue coating correlated with improvements in assessed variables related to oral health, such as oral malodor and plaque accumulation. Furthermore, there was significantly less microbiota diversity in supragingival plaque at 8 weeks in the test group than in the placebo group and low microbiota diversity correlated with improvements in assessed variables related to oral health. These results suggest that LF and LPO-containing tablets promote a shift from a highly diverse and gram-negative-dominated to a gram-positive-dominated community in the microbiota of supragingival plaque and tongue coating. This microbial shift may contribute to improvements in oral health, including oral malodor and state of the gingiva.
- Published
- 2017
9. A comparison of vasodilation mode among selexipag (NS-304; [2-{4-[(5,6-diphenylpyrazin-2-yl)(isopropyl)amino]butoxy}-N-(methylsulfonyl)acetamide]), its active metabolite MRE-269 and various prostacyclin receptor agonists in rat, porcine and human pulmonary arteries
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Michiko Oka, Junko Homan, Kohji Murakami, Kazuya Kuramoto, Keiichi Kuwano, Keiji Kosugi, Koyuki Tajima, and Chiaki Fuchikami
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0301 basic medicine ,Agonist ,Male ,medicine.medical_specialty ,medicine.drug_class ,Swine ,Vasodilator Agents ,Vasodilation ,030204 cardiovascular system & hematology ,Selexipag ,Acetates ,Pulmonary Artery ,Nitric Oxide ,Receptors, Epoprostenol ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Internal medicine ,medicine.artery ,Acetamides ,medicine ,Cyclic AMP ,Animals ,Humans ,Receptor ,Prostacyclin receptor ,Pharmacology ,Rats ,030104 developmental biology ,Endocrinology ,chemistry ,Pyrazines ,Pulmonary artery ,Endothelium, Vascular ,Iloprost ,medicine.drug ,Treprostinil - Abstract
Selexipag (NS-304; [2-{4-[(5,6-diphenylpyrazin-2-yl)(isopropyl)amino]butoxy}-N- (methylsulfonyl)acetamide]) is a novel, orally available non-prostanoid prostacyclin receptor (IP receptor) agonist that has recently been approved for the treatment of pulmonary arterial hypertension (PAH). We examined the effect of the active metabolite of selexipag, MRE-269, and IP receptor agonists that are currently available as PAH therapeutic drugs on the relaxation of rat, porcine and human pulmonary artery. cAMP formation in human pulmonary artery smooth muscle cells was induced by all test compounds (MRE-269, epoprostenol, iloprost, treprostinil and beraprost sodium) and suppressed by IP receptor antagonists (CAY10441 and 2-[4-(1H-indol-4-yloxymethyl)-benzyloxycarbonylamino]-3-phenyl-propionic acid). MRE-269 induced endothelium-independent vasodilation of rat extralobar pulmonary artery (EPA). In contrast, endothelial denudation or the addition of a nitric oxide synthase inhibitor markedly attenuated the vasodilation of EPA induced by epoprostenol, treprostinil and beraprost sodium but not iloprost. The vasorelaxant effects of MRE-269 on rat small intralobar pulmonary artery (SIPA) and EPA were the same, while the other IP receptor agonists induced less vasodilation in SIPA than in EPA. Furthermore, a prostaglandin E receptor 3 antagonist enhanced the vasodilation induced by all IP receptor agonists tested except MRE-269. We also investigated the relaxation induced by IP receptor agonists in pulmonary arteries from non-rodent species and found similar vasodilation modes in porcine and human as in rat preparations. These results suggest that MRE-269, in contrast to other IP receptor agonists, works as a selective IP receptor agonist, thus leading to pronounced vasorelaxation of rat, porcine and human pulmonary artery.
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- 2016
10. Condensation Energy Density Properties of Ba-122 Pnictide Superconductor with Columnar Defects Introduced by Heavy-ion Irradiation
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Masaru Kiuchi, Kazuaki Myose, J. Ge, Tsuyoshi Tamegai, Kohji Murakami, Baorong Ni, Teruo Matsushita, Yasuyuki Nakajima, and Edmund Soji Otabe
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Superconductivity ,Materials science ,Flux pinning ,Number density ,Condensed matter physics ,Condensation energy density ,Condensation ,Physics and Astronomy(all) ,Heavy-ion irradiation ,Condensed Matter::Superconductivity ,Cuprate ,Irradiation ,Ba-122 single crystal ,Anisotropy ,Pinning force - Abstract
It is important to investigate condensation energy density of superconducting material, since it determines its pinning property. According to the heavy-ion irradiation, the columnar defects are introduced to the superconductor which act as the strong pinning centers, and the critical current density is enhanced by the new pinning centers. Since the number density and the size of the columnar defects can be observed, the condensation energy density is estimated by using the pinning summation theory. In the present study, we prepared Ba(Fe0.93Co0.07)2As2 (Ba-122) pnictide superconductors by self-flux method. 200 MeV Au ions were irradiated into the specimens along c-axis and the matching field was 2 T. After the irradiation, the critical current density was 6 times larger than that before the irradiation. The estimated condensation energy density is in the order of 104 J/m3 and is slightly smaller than those of cuprate superconductors. The temperature dependence is similar to that of YBa2Cu3Ox oxide superconductors, since the anisotropy parameter is small in Ba-122. Therefore, it is expected to use Ba-122 at high temperatures near the critical temperature., Superconductivity Centennial Conference 2011 - EUCAS–ISEC–ICMC, Sep 18 - 23, 2011, Hague, Netherlands
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- 2012
- Full Text
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11. Discovery of a novel class of 1,3-dioxane-2-carboxylic acid derivatives as subtype-selective peroxisome proliferator-activated receptor α (PPARα) agonists
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Makoto Todo, Tomiyoshi Aoki, Tetsuo Asaki, Shinji Ohmachi, Kenji Kuwabara, Kohji Murakami, Yukiteru Sugiyama, and Taisuke Hamamoto
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Blood Glucose ,Agonist ,Peroxisome proliferator-activated receptor gamma ,Magnetic Resonance Spectroscopy ,medicine.drug_class ,Lipoproteins ,Carboxylic acid ,Clinical Biochemistry ,Pharmaceutical Science ,Peroxisome proliferator-activated receptor ,Mice, Inbred Strains ,Biochemistry ,Dioxanes ,Mice ,Structure-Activity Relationship ,Drug Discovery ,medicine ,Animals ,Humans ,Hypoglycemic Agents ,Structure–activity relationship ,PPAR alpha ,Receptor ,Oxazoles ,Molecular Biology ,Triglycerides ,Hypolipidemic Agents ,chemistry.chemical_classification ,Molecular Structure ,Chemistry ,Organic Chemistry ,Disease Models, Animal ,Diabetes Mellitus, Type 2 ,Nuclear receptor ,Molecular Medicine ,Peroxisome proliferator-activated receptor alpha - Abstract
A new series of 1,3-dioxane-2-carboxylic acid derivatives was synthesized and evaluated for agonist activity at human peroxisome proliferator-activated receptor (PPAR) subtypes. Structure-activity relationship studies led to the identification of 2-methyl-c-5-[4-(5-methyl-2-phenyl-1,3-oxazol-4-yl)butyl]-1,3-dioxane-r-2-carboxylic acid 4b as a potent PPARalpha agonist with high subtype selectivity at human receptor subtypes. This compound exhibited a substantial hypolipidemic effect in type 2 diabetic KK-A(y) mice.
- Published
- 2008
12. Relationship between skeletal muscle mass and swallowing function in patients with Alzheimer's disease
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Daisuke, Takagi, Hirohiko, Hirano, Yutaka, Watanabe, Ayako, Edahiro, Yuki, Ohara, Hideyo, Yoshida, Hunkyung, Kim, Kohji, Murakami, and Shouji, Hironaka
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Aged, 80 and over ,Male ,Nutritional Status ,Organ Size ,Prognosis ,Severity of Illness Index ,Statistics, Nonparametric ,Body Mass Index ,Cohort Studies ,Disability Evaluation ,Logistic Models ,Japan ,Alzheimer Disease ,Multivariate Analysis ,Confidence Intervals ,Disease Progression ,Odds Ratio ,Humans ,Female ,Deglutition Disorders ,Muscle, Skeletal ,Geriatric Assessment ,Retrospective Studies - Abstract
The present study verified the hypothesis that decreased skeletal muscle in older adults with Alzheimer's disease is related to Alzheimer's disease progression and decreased oral or swallowing function.We investigated 232 patients with Alzheimer's disease (31 men, 201 women, average age 85.4 ± 5.9 years) in two regions in Japan. The patients provided basic information (sex and age), and were assessed for skeletal muscle index, dementia severity (clinical dementia rating), activities of daily living, nutritional status, oral status and swallowing function.Stratification by clinical dementia rating was as follows: clinical dementia rating 0.5:21 patients (9.0%), clinical dementia rating 1:85 patients (36.6%), clinical dementia rating 2:88 patients (37.9%) and clinical dementia rating 3:38 patients (16.3%). Alzheimer's disease severity was significantly related to skeletal muscle index. Logistic regression analysis showed that clinical dementia rating 3 (odds ratio 11.68, 95% confidence interval 4.52-30.20), body mass index 18.5 (odds ratio 3.18, 95% confidence interval 1.27-8.00), calf circumference30.5 cm (odds ratio 9.33, 95% confidence interval 2.01-43.27) and poor swallowing function (odds ratio 4.93, 95% confidence interval 1.10-22.04) had a significant effect on decreased skeletal muscle index.Therefore, decreased skeletal muscle mass in patients with Alzheimer's disease requires strategies to manage swallowing dysfunction. Geriatr Gerontol Int 2016; 17: 402-409.
- Published
- 2015
13. Relationship between swallowing function and the skeletal muscle mass of older adults requiring long-term care
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Hirohiko Hirano, Daisuke Takagi, Hunkyung Kim, Yutaka Watanabe, Ayako Edahiro, Yuki Ohara, Hideyo Yoshida, Shouji Hironaka, and Kohji Murakami
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Male ,medicine.medical_specialty ,Nutritional Status ,Motor Activity ,Body Mass Index ,Swallowing ,Japan ,Tongue ,Risk Factors ,Internal medicine ,Medicine ,Humans ,Risk factor ,Muscle, Skeletal ,Aged ,Aged, 80 and over ,Univariate analysis ,business.industry ,Odds ratio ,Dysphagia ,Long-Term Care ,Confidence interval ,Deglutition ,medicine.anatomical_structure ,Physical therapy ,Female ,medicine.symptom ,business ,Deglutition Disorders ,Body mass index - Abstract
Aim The present study investigated the risk factors for dysphagia among older adults who require long-term care, and also examined their systemic decrease in skeletal muscle mass. Methods We evaluated 399 people who required long-term care and who were residing in Omori town, Yokote city, Akita prefecture, Japan. We then analyzed data from 255 participants who had complete information available regarding their sex, age, case history (stroke, Parkinson's disease and dementia), Barthel Index, Skeletal Muscle Mass Index, oral function test and modified water swallowing test results. Participants' water swallowing test results were used to create groups with good or poor swallowing function, and a univariate analysis was carried out for each parameter. Parameters with a P-value of
- Published
- 2015
14. DHCR24-Knockout Embryonic Fibroblasts Are Susceptible to Serum Withdrawal-Induced Apoptosis Because of Dysfunction of Caveolae and Insulin-Akt-Bad Signaling
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Hisao Seo, Takahide Kaji, Takehisa Ishii, Xiuli Lu, David Zadworny, Kohji Murakami, Taemi Yoshida, Sachiko Ohmori, Xia Cao, and Fukushi Kambe
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Oxidoreductases Acting on CH-CH Group Donors ,medicine.medical_specialty ,Programmed cell death ,Cell Survival ,medicine.medical_treatment ,Caveolin 1 ,Apoptosis ,Nerve Tissue Proteins ,Caveolae ,Culture Media, Serum-Free ,Mice ,chemistry.chemical_compound ,Endocrinology ,Desmosterol ,Internal medicine ,medicine ,Animals ,Insulin ,Protein kinase B ,Cells, Cultured ,Mice, Knockout ,biology ,beta-Cyclodextrins ,Receptor, Insulin ,Mice, Inbred C57BL ,Insulin receptor ,Cholesterol ,chemistry ,biology.protein ,bcl-Associated Death Protein ,Signal transduction ,Proto-Oncogene Proteins c-akt ,Signal Transduction - Abstract
The DHCR24 gene encodes an enzyme catalyzing the last step of cholesterol biosynthesis, the conversion of desmosterol to cholesterol. To elucidate the physiological significance of cholesterol biosynthesis in mammalian cells, we investigated proliferation of mouse embryonic fibroblasts (MEFs) prepared from DHCR24(-/-) mice. Both DHCR24(-/-) and wild-type MEFs proliferated in the presence of serum in culture media. However, the inhibition of external cholesterol supply by serum withdrawal induced apoptosis of DHCR24(-/-) MEFs, which was associated with a marked decrease in the intracellular and plasma membrane cholesterol levels, Akt inactivation, and Bad dephosphorylation. Insulin is an antiapoptotic factor capable of stimulating the Akt-Bad cascade, and its receptor (IR) is enriched in caveolae, cholesterol-rich microdomains of plasma membrane. We thus analyzed the association of IR and caveolae in the cholesterol-depleted MEFs. Subcellular fractionation and immunocytochemical analyses revealed that the IR and caveolin-1 contents were markedly reduced in the caveolae fraction of the MEFs, suggesting the disruption of caveolae, and that large amounts of IR were present apart from caveolin-1 on plasma membrane, indicating the uncoupling of IR with caveolae. Consistent with these findings, insulin-dependent phosphorylations of insulin receptor substrate-1, Akt, and Bad were impaired in the cholesterol-depleted MEFs. However, this impairment was partial because treatment of the MEFs with insulin restored Akt activation and prevented apoptosis. Cholesterol supply also prevented apoptosis. These results demonstrate that the cellular cholesterol biosynthesis is critical for the activation and maintenance of the Akt-Bad cell survival cascade in response to growth factors such as insulin.
- Published
- 2006
15. Prognosis of Breast Cancer Patients Treated with Sentinel Node Biopsy in Japan
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Takahiro Hasebe, Noriaki Wada, Satoshi Ebihara, Atsushi Ochiai, Shigeru Imoto, and Kohji Murakami
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Adult ,Cancer Research ,medicine.medical_specialty ,Breast Neoplasms ,Breast cancer ,Japan ,Biopsy ,medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,Survival rate ,Retrospective Studies ,Frozen section procedure ,medicine.diagnostic_test ,Sentinel Lymph Node Biopsy ,business.industry ,General surgery ,Axillary Lymph Node Dissection ,General Medicine ,Sentinel node ,Prognosis ,medicine.disease ,Survival Rate ,Axilla ,medicine.anatomical_structure ,Oncology ,Lymphatic Metastasis ,Lymph Node Excision ,Female ,Lymph Nodes ,Radiology ,Lymph ,business - Abstract
Background: Sentinel node biopsy predicts accurate pathological nodal staging. The survival of node-negative breast cancer patients should be evaluated between the patients treated with sentinel node biopsy alone and those treated with axillary lymph node dissection. Methods: Ninety-seven patients with negative axillary nodes underwent sentinel node biopsy immediately followed by axillary lymph node dissection between January 1998 and June 1999 (the ALND group). Since then, if sentinel lymph nodes were negative on the frozen-section diagnosis, 112 patients underwent sentinel node biopsy alone without axillary lymph node dissection between July 1999 and December 2000 (the SNB group). We retrospectively observed the outcome of the two study groups. Results: Median follow-up was 52 months in all patients. Relapse-free survival rates at 3 years in the ALND and SNB groups were 94% and 93%, respectively. Five of the 112 patients in the SNB group had overt axillary metastases. Three of them with axillary metastases alone were treated with delayed axillary lymph node dissection. These three patients have been free of other events for 3 years after local salvage treatment. Conclusions: Sentinel node biopsy will emerge as a standard method to diagnose axillary nodal staging for clinically node-negative breast cancer patients.
- Published
- 2004
16. Structure of asparagine-linked oligosaccharides of an aspartic proteinase from the zygomycete fungus Rhizomucor pusillus
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Sueharu Horinouch, Teruhiko Beppu, Kohji Murakami, and Kyoko Takeuchi
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Glycosylation ,Stereochemistry ,Molecular Sequence Data ,Oligosaccharides ,Mannose ,Microbiology ,Rhizomucor pusillus ,chemistry.chemical_compound ,Residue (chemistry) ,Carbohydrate Conformation ,Aspartic Acid Endopeptidases ,Asparagine ,Chromatography, High Pressure Liquid ,chemistry.chemical_classification ,biology ,Glycopeptides ,Oligosaccharide ,biology.organism_classification ,Rhizomucor ,Carbohydrate Sequence ,Biochemistry ,chemistry ,Mucor ,Carbohydrate conformation - Abstract
The zygomycete fungus Rhizomucor pusillus (previously called Mucor pusillus) secretes an aspartic proteinase containing two asparagine-linked, high-mannose type oligosaccharide chains at Asn79 and Asn188. For structural elucidation of the carbohydrate moieties, the protein was divided into two portions, an N-terminal portion containing Asn79 and a C-terminal portion containing Asn188, by a specific autocatalytic cleavage under alkaline conditions. Each of the asparagine-linked oligosaccharides was then released by peptide-N-glycosidase F digestion and pyridylaminated with a fluorescent reagent, 2-aminopyridine, at the reducing end. High-performance liquid chromatography analyses showed that the structure of the asparagine-linked oligosaccharide chain attached to residue Asn79 was Man5 GlcNAc2, and that bound to residue Asn188 was Man5 GlcNAc2 and Man5GlcNAc2. These observations suggest that the processing of mannose residues in asparagine-linked oligosaccharides in the Golgi apparatus of Rhizomucor resembles that in mammalian cells.
- Published
- 1998
17. Effects of Thermal Aging on Leucoemeraldine Form of Polyaniline
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Kohji Murakami and Hiroshi Tsubakihara
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chemistry.chemical_compound ,Materials science ,Polymers and Plastics ,chemistry ,Materials Science (miscellaneous) ,Polyaniline ,Chemical Engineering (miscellaneous) ,Thermal aging ,Composite material ,General Environmental Science - Abstract
還元脱ドープ状態のポリアニリンを種々の温度で大気中熱処理し, その電子, 分子構造を可視紫外, 赤外吸収スペクトルから推定した. 190℃までの熱処理では酸化により比較的短いキノンジイミン構造をポリアニリン分子内につくる. 高温熱処理ではイミンはその数を減少し, 残ったイミンも分子鎖内, 鎖間結合のもとで不活発となる. このことが硫酸水溶液に浸漬したときのドーピングされにくさの原因であると考えられる.
- Published
- 1997
18. Blockade of voltage-gated calcium channel Cav1.2 and α1-adrenoceptors increases vertebral artery blood flow induced by the antivertigo agent difenidol
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Asami Hashino, Kumiko Noda, Koyuki Tajima, Naoki Inoue, Michiko Oka, Kohji Murakami, Hisashi Fukui, and Chiaki Fuchikami
- Subjects
Male ,medicine.medical_specialty ,Calcium Channels, L-Type ,medicine.drug_class ,Calcium channel blocker ,CHO Cells ,Cav1.2 ,Calcium in biology ,Phentolamine ,Cricetulus ,Dogs ,Nifedipine ,Piperidines ,Internal medicine ,Cricetinae ,Receptors, Adrenergic, alpha-1 ,medicine ,Animals ,Humans ,Phenylephrine ,Vertebral Artery ,Pharmacology ,biology ,Voltage-dependent calcium channel ,Chemistry ,Calcium channel ,Calcium Channel Blockers ,Rats ,Endocrinology ,HEK293 Cells ,biology.protein ,Adrenergic alpha-1 Receptor Antagonists ,Vertigo ,Blood Flow Velocity ,medicine.drug - Abstract
Difenidol (1,1-diphenyl-4-piperidino-1-butanol hydrochloride) is an effective drug for the treatment of vertigo and dizziness. This drug is known to improve the blood flow in vertebral arteries, though the precise mechanism underlying this action remains unclear. In the present study, we investigated the effect of difenidol on voltage-gated calcium channel Ca(v)1.2 and α(1)-adrenoceptor subtypes that regulate the intracellular calcium concentration ([Ca(2+)](i)), as well as their possible involvement in the action of difenidol on vertebral artery relaxation and blood flow in dogs. In vitro binding assays demonstrated that difenidol at micromolar concentrations bound to the α(1A)-, α(1B)- and α(1D)-adrenoceptor subtypes. Difenidol inhibited the phenylephrine-induced increase in [Ca(2+)](i) in Chinese hamster ovary cells expressing human α(1A)-, α(1B)- or α(1D)-adrenoceptor subtypes with similar IC(50) values in the low micromolar range. In an electrophysiological assay, difenidol inhibited L-type calcium channel (Ca(v)1.2 subunit). In dogs, i.v. difenidol preferentially enhanced vertebral over femoral arterial blood flow. Phenylephrine and potassium induced contraction of dog vertebral arterial rings, and difenidol inhibited this action. Inhibition of phenylephrine-induced contraction by difenidol was mimicked by the α(1)-adrenoceptor antagonist phentolamine, the α(1A)-adrenoceptor antagonist RS 17,053 (N-[2-(2-cyclopropylmethoxyphenoxy)ethyl]-5-chloro-α,α-dimethyl-1H-indole-3-ethanamine hydrochloride) and the α(1D)-adrenoceptor antagonist BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4,5]decane-7,9-dione dihydrochloride). In addition, the L-type calcium channel blocker nifedipine, like difenidol, attenuated the potassium-induced contraction. These findings suggest that the difenidol-induced increase in vertebral arterial blood flow may be due to vascular relaxation mediated by mixed blocking actions at α(1)-adrenoceptors and voltage-gated calcium channel Ca(v)1.2.
- Published
- 2012
19. Characterization of an Aspartic Proteinase of Mucor pusillus expressed in Aspergillus oryzae
- Author
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Kohji Murakami, Jun-ichi Aikawa, Teruhiko Beppu, and Sueharu Horinouchi
- Subjects
Glycosylation ,Aspergillus oryzae ,Blotting, Western ,Molecular Sequence Data ,Mannose ,Microbiology ,law.invention ,chemistry.chemical_compound ,Transformation, Genetic ,law ,Gene expression ,Genetics ,Aspartic Acid Endopeptidases ,Cloning, Molecular ,DNA, Fungal ,Molecular Biology ,Southern blot ,chemistry.chemical_classification ,Base Sequence ,biology ,food and beverages ,Protoplast ,biology.organism_classification ,Recombinant Proteins ,Blotting, Southern ,Enzyme ,chemistry ,Biochemistry ,Mucor ,Recombinant DNA ,Carbohydrate Metabolism - Abstract
The aspartic proteinase (MPP) gene from the zygomycete fungus Mucor pusillus was introduced into an ascomycete fungus, Aspergillus oryzae, by protoplast transformation using the nitrate reductase (niaD) gene as the selective marker. Southern blot analysis indicated that the MPP gene was integrated into the resident niaD locus at a copy number of 1-2. MPP secreted by the recombinant A. oryzae was correctly processed but was more highly glycosylated than that produced in the original M. pusillus strain. Treatment with endo-beta-N-acetylglucosaminidase H and analysis of the carbohydrate composition of the secreted MPP revealed that the extra glycosylation of the MPP secreted by the recombinant A. oryzae was due to altered processing of mannose residues. The extra glycosylation of MPP affected its enzyme properties including its milk-clotting and proteolytic activities.
- Published
- 1993
20. Isolation and Characterization of the Alkaline Protease Gene ofAspergillus oryzae
- Author
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Hiroshi Motai, Yutaka Ishida, Hiroki Tatsumi, Eiichi Nakano, Atsushi Masaki, Hirofumi Arimura, Haruhide Kawabe, Seiji Murakami, and Kohji Murakami
- Subjects
biology ,TATA box ,CAAT box ,Nucleic acid sequence ,biology.organism_classification ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Primer extension ,genomic DNA ,Biochemistry ,Aspergillus oryzae ,Coding region ,General Agricultural and Biological Sciences ,Gene - Abstract
The genomic DNA for the alkaline protease (Alp) of the fungus Aspergillus oryzae was isolated using synthetic oligonucleotides as hydridization probes, and the complete nucleotide sequence was identified. The Alp gene is 1374 nucleotides long and contains three introns, one of which is in the pro region and two in the mature coding region. Sequences related to the TATA box (TATAAAT) and the CAAT box (CCAAAT) were found in the 5'-noncoding region. Primer extension analysis showed that three transcriptional start points are present.
- Published
- 1991
21. Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease
- Author
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Hiroki Tatsumi, Ryohei Tsuji, Atusi Masaki, Kohji Murakami, Eiichi Nakano, Haruhide Kawabe, Hiroshi Motai, Hirofumi Arimura, Seiji Murakami, and Yutaka Ishida
- Subjects
Sequence analysis ,Aspergillus oryzae ,medicine.medical_treatment ,Genes, Fungal ,Molecular Sequence Data ,Saccharomyces cerevisiae ,Gene Expression ,Transformation, Genetic ,Sequence Homology, Nucleic Acid ,Complementary DNA ,Genetics ,medicine ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,chemistry.chemical_classification ,Protease ,Base Sequence ,biology ,Nucleic acid sequence ,Metalloendopeptidases ,biology.organism_classification ,Molecular biology ,Yeast ,Amino acid ,Biochemistry ,chemistry ,Plasmids - Abstract
The neutral protease II (NpII) from Aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. To elucidate its structure, we isolated a full-length cDNA clone for NpII. Sequence analysis reveals that NpII has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. As compared with other microbial metalloproteases, NpII is found to be unique in that it shares only a limited homology with them around two zinc ligand His residues and that the positions of the other zinc ligand (Glu) and the active site (His) cannot be established by homology. When a plasmid designed to express the prepro NpII cDNA was introduced into Saccharomyces cerevisiae and the transformant was cultured in YPD medium (2% glucose, 2% polypeptone, 1% yeast extract), it secreted a proNpII. However, in a culture of the same medium containing 0.2 mM ZnCl2, it secreted a mature NpII with a specific activity and N-terminus identical to those of native NpII. This observation suggests that either an autoproteolytic activity or a yeast protease effected the processing.
- Published
- 1991
22. Secretory Expression of the Human Serum Albumin Gene in the Yeast, Saccharomyces cerevisiae
- Author
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Ken Okabayashi, Hirabayashi Kazumoto, Yukimitsu Nakagawa, Shimizu Miho, Takao Ohmura, Masami Miura, Haruhide Kawabe, Yutaka Ishida, Hideyuki Ohi, Naofumi Hayasuke, Akinori Sumi, Atsushi Masaki, Kohji Murakami, Hajime Horii, and Hitoshi Minamino
- Subjects
Genetic Vectors ,Molecular Sequence Data ,Saccharomyces cerevisiae ,Serum albumin ,Gene Expression ,Biochemistry ,Plasmid ,Complementary DNA ,Gene expression ,medicine ,Humans ,Prealbumin ,Amino Acid Sequence ,Cloning, Molecular ,Promoter Regions, Genetic ,Molecular Biology ,Serum Albumin ,Base Sequence ,biology ,Promoter ,DNA ,General Medicine ,Human serum albumin ,biology.organism_classification ,Molecular biology ,Yeast ,body regions ,embryonic structures ,biology.protein ,Plasmids ,medicine.drug - Abstract
We have fused a cDNA gene encoding mature human serum albumin (HSA) to several secretory leader-encoding sequences. The hybrid genes were cloned into an episomal vector under the control of several yeast promoters and then introduced into yeast cells. The GAL1 promoter in combination with either the native HSA pre-sequence or a modified HSA pre-sequence gave the highest production of immunoreactive HSA, 90 mg/liter being reached in a shake flask culture. The invertase pre-sequence, the mating factor alpha 1 prepro-sequence, and the modified HSA pre-sequence directed accurate processing. In contrast, the chicken lysozyme pre-sequence and the native HSA pre-sequence directed incorrect processing. Episomal vectors were unstable within the host cells under non-selective culture conditions. To improve the plasmid stability, the hybrid genes were incorporated into an integrative vector. Transformants carrying multicopies of the plasmid integrated at the LEU2 locus stably secreted HSA. The highest yield of 65 mg/liter in a shake flask culture was obtained with the combination of the yeast glyceraldehyde-3-phosphate dehydrogenase promoter and the modified HSA pre-sequence. By constructing transformed strains containing multicopies of plasmids integrated at both the chromosome LEU2 and HIS4 loci, we have obtained a stable strain that continuously secretes as much as 85 mg HSA per liter of culture medium.
- Published
- 1991
23. Isolation and Characterization of the Alkaline Protease Gene of Aspergillus oryzae
- Author
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Kohji MURAKAMI, Yutaka ISHIDA, Atsushi MASAKI, Hiroki TATSUMI, Seiji MURAKAMI, Eiichi NAKANO, Hiroshi MOTAI, Haruhide KAWABE, and Hirofumi ARIMURA
- Subjects
General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology - Published
- 1991
24. Secretion ofAspergillus oryzaeAlkaline Protease in an Osmophilic Yeast,Zygosaccharomyces rouxii
- Author
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Yoshihiro Ogawa, Hiroki Tatsumi, Seiji Murakami, Yutaka Ishida, Kohji Murakami, Atsushi Masaki, Haruhide Kawabe, Hirofumi Arimura, Eiichi Nakano, Hiroshi Motai, and Akio Toh-e
- Subjects
General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology - Published
- 1990
25. Secretion of Aspergillus oryzae alkaline protease in an osmophilic yeast, Zygosaccharomyces rouxii
- Author
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Seiji Murakami, Haruhide Kawabe, Yutaka Ishida, Akio Toh-e, Atsushi Masaki, Eiichi Nakano, Kohji Murakami, Hiroki Tatsumi, Hirofumi Arimura, Hiroshi Motai, and Yoshihiro Ogawa
- Subjects
musculoskeletal diseases ,chemistry.chemical_classification ,Expression vector ,biology ,musculoskeletal, neural, and ocular physiology ,food and beverages ,Dehydrogenase ,Fungi imperfecti ,musculoskeletal system ,biology.organism_classification ,General Biochemistry, Genetics and Molecular Biology ,Yeast ,Plasmid ,Enzyme ,stomatognathic system ,Aspergillus oryzae ,Biochemistry ,chemistry ,Complementary DNA ,General Agricultural and Biological Sciences - Abstract
To produce Aspergillus oryzae alkaline protease (Alp) in an osmophilic yeast Zygosaccharomyces rouxii, we constructed an expression plasmid consisting of the Z. rouxii glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter, the prepro-Alp cDNA of A. oryzae, the whole sequence of Z. rouxii plasmid pSR1, and the G418 resistant gene. The resulting plasmid, when introduced into Z. rouxii cells, directed the secretion of a large amount (about 300 mg/l) of Alp into the culture medium. The N-terminus and specific activity of the enzyme were identical to those of A. oryzae Alp.
- Published
- 1990
26. Structure-activity studies on 1,3-dioxane-2-carboxylic acid derivatives, a novel class of subtype-selective peroxisome proliferator-activated receptor alpha (PPARalpha) agonists
- Author
-
Makoto Todo, Yukiteru Sugiyama, Tomiyoshi Aoki, Shinji Ohmachi, Kohji Murakami, Tetsuo Asaki, Kenji Kuwabara, and Taisuke Hamamoto
- Subjects
Agonist ,medicine.drug_class ,Carboxylic acid ,Clinical Biochemistry ,Pharmaceutical Science ,Peroxisome proliferator-activated receptor ,Biochemistry ,Dioxanes ,chemistry.chemical_compound ,Transactivation ,Mice ,Structure-Activity Relationship ,Drug Discovery ,medicine ,Structure–activity relationship ,Animals ,Combinatorial Chemistry Techniques ,Humans ,PPAR alpha ,Molecular Biology ,Oxazoles ,Oxazole ,chemistry.chemical_classification ,Molecular Structure ,Cholesterol ,Organic Chemistry ,chemistry ,Nuclear receptor ,Molecular Medicine ,lipids (amino acids, peptides, and proteins) - Abstract
A series of 1,3-dioxane carboxylic acid derivatives was synthesized and evaluated for human PPAR transactivation activity. Structure-activity relationships on the phenyloxazole moiety of the lead compound 3 revealed that the introduction of small hydrophobic substituents at the 4-position of the terminal phenyl ring increased the PPARalpha agonist activity, and that the oxazole heterocycle was essential to the maintenance of both potency and PPARalpha subtype-selectivity. This investigation led to the identification of 14d (NS-220) and 14i as highly potent and selective human PPARalpha agonists. In KK-A(y) type 2 diabetic mice, these compounds significantly lowered plasma triglyceride and very-low-density plus low-density lipoprotein cholesterol levels while simultaneously raising HDL cholesterol levels. Our results suggest that highly potent and subtype-selective PPARalpha agonists will be promising drugs for the treatment of metabolic disorders in type 2 diabetes.
- Published
- 2007
27. DHCR24 gene knockout mice demonstrate lethal dermopathy with differentiation and maturation defects in the epidermis
- Author
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Fukushi Kambe, Sachiko Ohmori, Hisao Seo, Yoshiharu Murata, Kazuko Maki, David Zadworny, Yoshiko Takagishi, Michiyo Yamamoto, Kohji Murakami, Shizu Hayasaka, Takahide Kaji, and Rusella Mirza
- Subjects
Keratinocytes ,Pathology ,medicine.medical_specialty ,Oxidoreductases Acting on CH-CH Group Donors ,Caveolin 1 ,Apoptosis ,Nerve Tissue Proteins ,Dermatology ,Biology ,Fatty Acids, Nonesterified ,Ceramides ,Biochemistry ,Skin Diseases ,Permeability ,chemistry.chemical_compound ,Mice ,Desmosterol ,Keratin ,medicine ,Animals ,Molecular Biology ,Gene knockout ,Cell Proliferation ,Skin ,chemistry.chemical_classification ,Mice, Knockout ,Epidermis (botany) ,integumentary system ,Cell Differentiation ,Cell Biology ,medicine.disease ,Molecular biology ,Immunohistochemistry ,Desmosterolosis ,Mice, Inbred C57BL ,Microscopy, Electron ,Cholesterol ,chemistry ,Knockout mouse ,Loricrin ,Keratins ,Filaggrin - Abstract
Desmosterolosis is an autosomal recessive disorder due to mutations in the 3beta-hydroxysterol-Delta24 reductase (DHCR24) gene that encodes an enzyme catalyzing the conversion of desmosterol to cholesterol. To date, only two patients have been reported with severe developmental defects including craniofacial abnormalities and limb malformations. We employed mice with targeted disruption of DHCR24 to understand the pathophysiology of desmosterolosis. All DHCR24-/- mice died within a few hours after birth. Their skin was wrinkleless and less pliant, leading to restricted movement and inability to suck (empty stomach). DHCR24 gene was expressed abundantly in the epidermis of control but not of DHCR24-/- mice. Accordingly, cholesterol was not detected whereas desmosterol was abundant in the epidermis of DHCR24-/- mice. Skin histology revealed thickened epidermis with few and smaller keratohyaline granules. Aberrant expression of keratins such as keratins 6 and 14 suggested hyperproliferative hyperkeratosis with undifferentiated keratinocytes throughout the epidermis. Altered expression of filaggrin, loricrin, and involcrin were also observed in the epidermis of DHCR24-/-. These findings suggested impaired skin barrier function. Indeed, increased trans-epidermal water loss and permeability of Lucifer yellow were observed in DHCR24-/- mice. DHCR24 thus plays crucial role for skin development and its proper function.
- Published
- 2006
28. A novel selective peroxisome proliferator-activated receptor alpha agonist, 2-methyl-c-5-[4-[5-methyl-2-(4-methylphenyl)-4-oxazolyl]butyl]-1,3-dioxane-r-2-carboxylic acid (NS-220), potently decreases plasma triglyceride and glucose levels and modifies lipoprotein profiles in KK-Ay mice
- Author
-
Tomiyoshi Aoki, Junichi Yano, Makoto Todo, Kohji Murakami, Masatoshi Murai, Tetsuo Asaki, and Kenji Kuwabara
- Subjects
Agonist ,Blood Glucose ,Male ,medicine.medical_specialty ,medicine.drug_class ,Lipoproteins ,Alpha (ethology) ,Peroxisome proliferator-activated receptor ,Receptors, Cytoplasmic and Nuclear ,Dioxanes ,Mice ,Fenofibrate ,Internal medicine ,Hyperlipidemia ,medicine ,Animals ,Humans ,Hypoglycemic Agents ,Receptor ,Oxazoles ,Cells, Cultured ,Triglycerides ,Hypolipidemic Agents ,Pharmacology ,chemistry.chemical_classification ,Bezafibrate ,Binding Sites ,medicine.disease ,Endocrinology ,chemistry ,Molecular Medicine ,medicine.drug ,Lipoprotein ,Transcription Factors - Abstract
2-Methyl-c-5-[4-[5-methyl-2-(4-methylphenyl)-4-oxazolyl]butyl]-1,3-dioxane-r-2-carboxylic acid (NS-220) was newly synthesized and demonstrated to be a novel potent peroxisome proliferator-activated receptor alpha (PPARalpha) agonist with high subtype selectivity. In cell-based reporter gene assays, the EC(50) values of NS-220 for human PPARalpha, PPARgamma, and PPARdelta were 1.9 x 10(-8), 9.6 x 10(-6), and >10(-4) M, respectively, and for mouse PPARalpha, PPARgamma, and PPARdelta were 5.5 x 10(-8), 3.3 x 10(-5), and >10(-4) M, respectively. In addition, [(3)H]NS-220 bound to the ligand-binding domain of human PPARalpha with a K(D) value of 1.85 x 10(-7) M. Fenofibric acid and bezafibrate showed weak agonist activity for PPARalpha (EC(50), 2-8 x 10(-5) M), with poor subtype selectivity. NS-220 (0.1-3 mg/kg p.o.) decreased plasma triglyceride levels in ddY mice in a dose-dependent manner, but its hypolipidemic activity was abolished in PPARalpha-deficient mice. In KK-A(y) mice, an animal model of type-2 diabetes, NS-220 (0.3-1 mg/kg p.o.; 4 days) and fenofibrate (100-300 mg/kg p.o.; 4 days) decreased plasma triglyceride and glucose levels in a dose-dependent manner. In a 2-week repeated administration test, NS-220 (0.3-1 mg/kg p.o.) decreased plasma glucose levels markedly without increasing in plasma insulin levels. Furthermore, NS-220 increased high-density lipoprotein levels and decreased triglyceride-rich lipoprotein levels. In conclusion, a newly synthesized dioxanecarboxylic acid derivative, NS-220, is a potent and highly selective PPARalpha agonist that ameliorates metabolic disorders in diabetic mice. These results strongly suggest that it will be a promising drug for the treatment of hyperlipidemia or metabolic disorders in type-2 diabetes.
- Published
- 2004
29. Kinetic analysis of enhanced thermal stability of an alkaline protease with engineered twin disulfide bridges and calcium-dependent stability
- Author
-
Kouichi Yamanouchi, Shigetoshi Sugio, Kohji Murakami, and Kazuo Ikegaya
- Subjects
Models, Molecular ,Protein Denaturation ,Hot Temperature ,Protein Conformation ,Aspergillus oryzae ,Enthalpy ,Kinetics ,chemistry.chemical_element ,Bioengineering ,Calcium ,Applied Microbiology and Biotechnology ,Ion ,Reaction rate ,Protein structure ,Enzyme Stability ,Thermal stability ,Cysteine ,Disulfides ,Chemistry ,Serine Endopeptidases ,Temperature ,Biochemistry ,Biophysics ,Thermodynamics ,Genetic Engineering ,Biotechnology ,Half-Life ,Protein Binding - Abstract
The thermal stability of a cysteine-free alkaline protease (Alp) secreted by the eukaryote Aspergillus oryzae was improved both by the introduction of engineered twin disulfide bridges (Cys-69/Cys-101 and Cys-169/Cys-200), newly constructed as part of this study, and by the addition of calcium ions. We performed an extensive kinetic analysis of the increased thermal stability of the mutants as well as the role of calcium dependence. The thermodynamic activation parameters for irreversible thermal inactivation, the activation free energy (deltaG), the activation enthalpy (deltaH), and the activation entropy (deltaS) were determined from absolute reaction rate theory. The values of deltaH and deltaS were significantly and concomitantly increased as a result of introducing the twin disulfide bridges, for which the increase in the value of deltaH outweighed that of deltaS, resulting in significant increases in the value of deltaG. The enhancement of the thermal stability obtained by introducing the twin disulfide bridges is an example of the so-called low-temperature stabilization of enzymes. The stabilizing effect of calcium ions on wild-type Alp is similar to the results we obtained by introducing the engineered twin disulfide bridges.
- Published
- 2002
30. Pilot study on sentinel node biopsy in breast cancer
- Author
-
Hiroyoshi Fukukita, Noriyuki Moriyama, Shigeru Imoto, Kohji Murakami, and Hiroshi Ikeda
- Subjects
medicine.medical_specialty ,Biopsy ,Mammary gland ,Breast Neoplasms ,Pilot Projects ,Scintigraphy ,Indigo Carmine ,Lymphatic mapping ,Metastasis ,Breast cancer ,medicine ,Humans ,Lymph node ,Neoplasm Staging ,medicine.diagnostic_test ,business.industry ,General Medicine ,Sentinel node ,medicine.disease ,Surgery ,body regions ,medicine.anatomical_structure ,Oncology ,Axilla ,Female ,Radiology ,Lymph Nodes ,business - Abstract
Background and Objectives Sentinel node biopsy (SNB) in breast cancer using indigo carmine was started in January 1998, and this method has proved feasible and reliable. From our initial experience, sentinel lymph nodes (SLNs) were identified in 65 of 88 cases of breast cancer (74%). Methods Lymphatic mapping in breast cancer was assessed using radionuclide, technetium-99m human serum albumin or technetium-99m tin colloid. A pilot study on SNB with dye or a combined method was performed between August 1998 and January 1999. Results SLNs were identified in 55 of 59 cases (93%). False-negative SLNs were found in 2 cases. The sensitivity and accuracy in all cases were 92% and 96%. SLNs in 52 cases were also diagnosed by immediate frozen sectioning. The sensitivity and accuracy were 89% and 96%. Conclusions SNB in the combined method was the best way to identify SLNs in breast cancer. J. Surg. Oncol. 2000;73:130–133. © 2000 Wiley-Liss, Inc.
- Published
- 2000
31. Autoproteolytic Processing and Processing-dependent Secretion ofAspergillus oryzaeAlkaline Protease in Yeast
- Author
-
Hiroki Tatsumi, Seiji Murakami, Yoshihiro Ogawa, Atsushi Masaki, Yutaka Ishida, Kohji Murakami, Haruhide Kawabe, Hirofumi Arimura, Eiichi Nakano, and Hiroshi Motai
- Subjects
General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology - Published
- 1991
32. Autoproteolytic Processing and Processing-dependent Secretion of Aspergillus oryzae Alkaline Protease in Yeast
- Author
-
Eiichi Nakano, Kohji Murakami, Yutaka Ishida, Atsushi Masaki, Hirofumi Arimura, Hiroshi Motai, Yoshihiro Ogawa, Hiroki Tatsumi, Haruhide Kawabe, and Seiji Murakami
- Subjects
chemistry.chemical_classification ,biology ,Chemistry ,Alkaline protease ,Fungi imperfecti ,biology.organism_classification ,General Biochemistry, Genetics and Molecular Biology ,Yeast ,Microbiology ,Enzyme ,Aspergillus oryzae ,Biochemistry ,Secretion ,General Agricultural and Biological Sciences ,Gene - Published
- 1991
33. Substitution of amino acid residue in influenza A virus hemagglutinin affects recognition of sialyl-oligosaccharides containing N-glycolylneuraminic acid
- Author
-
Toshihiro Ito, Takashi Suzuki, Daisei Miyamoto, Tetsuya Toyoda, Yasuo Suzuki, Akira Ishihama, Yoshikuni Sugiyama, Masanobu Ohuchi, Kyoko Fukunaga, Makoto Kiso, Hiroyuki Masuda, Yoshihiro Kawaoka, Kazuya I.P.J. Hidari, Hiroshi Kida, Goh Horiike, and Kohji Murakami
- Subjects
Models, Molecular ,Sialoglycoproteins ,Molecular Sequence Data ,Biophysics ,Hemagglutinin (influenza) ,Biology ,medicine.disease_cause ,Biochemistry ,Microbiology ,chemistry.chemical_compound ,Viral Proteins ,Structural Biology ,Cell surface receptor ,N-Glycolylneuraminic acid ,Genetics ,Influenza A virus ,medicine ,Humans ,Amino Acid Sequence ,Hemagglutinin ,Sugar ,Molecular Biology ,chemistry.chemical_classification ,Sequence Homology, Amino Acid ,Sialyl-oligosaccharide ,Nucleic acid sequence ,Cell Biology ,Sialic acid ,Amino acid ,Hemagglutinins ,chemistry ,Carbohydrate Sequence ,biology.protein ,Neuraminic Acids ,Chromatography, Thin Layer ,Influenza virus ,Glycoconjugates - Abstract
Sialic acids are essential components of cell surface receptors used by influenza viruses. To determine the molecular mechanisms of viral recognition of two major species of sialic acids, N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc), we tested the binding reactivity of nine human H3 influenza A viruses to sialylglycolipids containing type II sugar chain and different molecular species of terminal sialic acids. All human H3 viruses tested except A/Memphis/1/71 bound both Neu5Ac and Neu5Gc. Nucleotide sequence analysis suggests that amino acids at 143, 155, and 158 are linked to the viral recognition of Neu5Gc.
- Published
- 1999
34. Mammary lymphoscintigraphy with various radiopharmaceuticals in breast cancer
- Author
-
Noriyuki Moriyama, Hiroyoshi Fukukita, Hiroshi Ikeda, Kohji Murakami, and Shigeru Imoto
- Subjects
Adult ,medicine.medical_specialty ,Pathology ,Time Factors ,Biopsy ,Sentinel lymph node ,Serum albumin ,Breast Neoplasms ,Breast cancer ,Parenchyma ,medicine ,Humans ,Radiology, Nuclear Medicine and imaging ,Radionuclide Imaging ,Technetium Tc 99m Aggregated Albumin ,Aged ,medicine.diagnostic_test ,biology ,business.industry ,Axillary Lymph Node Dissection ,Tin Compounds ,General Medicine ,Sentinel node ,Middle Aged ,medicine.disease ,body regions ,Technetium Compounds ,Lymphatic Metastasis ,biology.protein ,Technetium Tc 99m Pentetate ,Female ,Radiology ,Lymph ,Lymph Nodes ,Radiopharmaceuticals ,business - Abstract
Sentinel node biopsy (SNB) in breast cancer is a promising surgical technique that avoids unnecessary axillary lymph node dissection. To optimize lymphatic mapping with radiopharmaceuticals, mammary lymphoscintigraphy with 30-50 MBq of technetium-99m-diethylenetriamine pentaacetic acid human serum albumin (99mTc-HSAD), technetium-99m-human serum albumin (99mTc-HSA), or technetium-99m-tin colloid (99mTc-TC) were investigated in 69 cases of primary breast cancer. Dynamic early images were obtained during the first 30 or 40 minutes, and static delayed images were obtained 6 hours after tracer injection. Hot spots as sentinel lymph nodes (SLNs) appeared in 51 of 69 cases (74%): in early images in 27 cases and in delayed images in 24 cases. SLNs were visualized more frequently in 23 of the 26 cases (88%) treated with 99mTc-HSAD and in 21 of the 24 cases (88%) treated with 99mTc-HSA than in only 7 of the 19 cases (37%) treated with 99mTc-TC. In 26 of the 51 cases, SLNs were identified as faint spots in delayed images. There was a significant difference in the first appearance of SLNs on the lymphoscintiscan between 43 cases of dense breast parenchyma and 26 cases of fatty breast parenchyma. These results suggest that 99mTc-HSAD or 99mTc-HSA is acceptable for lymphatic mapping, but in cases which have faint spots in delayed images or fatty breast parenchyma, gamma probe-guided SNB may result in failure or misleading false-negative SLNs.
- Published
- 1999
35. A Mucor pusillus mutant defective in asparagine-linked glycosylation
- Author
-
Kohji Murakami, Jun-ichi Aikawa, M. Wada, Sueharu Horinouchi, and Teruhiko Beppu
- Subjects
Glycosylation ,1-Deoxynojirimycin ,Glycoside Hydrolases ,Mutant ,Oligosaccharides ,Regulatory Sequences, Nucleic Acid ,Microbiology ,chemistry.chemical_compound ,Glycoside hydrolase ,Asparagine ,Molecular Biology ,Mucor ,chemistry.chemical_classification ,biology ,Genetic Complementation Test ,Indolizines ,Oligosaccharide ,biology.organism_classification ,Molecular biology ,Pepsin A ,Enzyme ,chemistry ,Biochemistry ,Castanospermine ,Mutation ,Protein Processing, Post-Translational ,Research Article - Abstract
A Mucor pusillus mutant defective in asparagine-linked glycosylation was found in our stock cultures. This mutant, designated 1116, secreted aspartic proteinase (MPP) in a less-glycosylated form than that secreted by the wild-type strain. Analysis of enzyme susceptibility, lectin binding, and carbohydrate composition indicated that this mutant secreted three glycoforms of MPPs, one of which contained no carbohydrate; the other two had truncated asparagine-linked oligosaccharide chains such as Man0-1GlcNAc2. Further analysis using oligosaccharide processing inhibitors, such as castanospermine, 1-deoxynojirimycin and N-methyldeoxynojirimycin, suggested that MPPs in the mutant were glycosylated through a transfer of the truncated lipid-linked oligosaccharides, Man0-1GlcNAc2, to the MPP protein but not through an aberrant processing. In addition, genetic studies with forced primary heterokaryons indicated that the mutation in strain 1116 was recessive.
- Published
- 1994
36. Enhancement of the Thermostability of the Alkaline Protease from Aspergillus oryzae by Introduction of a Disulfide Bond
- Author
-
Seiji Murakami, Kohji Murakami, Yutaka Ishida, Hiroshi Motai, Haruhide Kawabe, Atsushi Masaki, Yoshihiro Ogawa, Hiroki Tatsumi, Eiichi Nakano, Naritoshi Sugio, Kazuo Takechi, and Kazuo Ikegaya
- Subjects
Hot Temperature ,Aspergillus oryzae ,Molecular Sequence Data ,Biology ,Applied Microbiology and Biotechnology ,Biochemistry ,Analytical Chemistry ,Drug Stability ,Amino Acid Sequence ,Disulfides ,Site-directed mutagenesis ,Molecular Biology ,Thermostability ,chemistry.chemical_classification ,Base Sequence ,Serine Endopeptidases ,Organic Chemistry ,Disulfide bond ,food and beverages ,Alkaline protease ,General Medicine ,biology.organism_classification ,Enzyme ,chemistry ,Biotechnology - Abstract
(1992). Enhancement of the Thermostability of the Alkaline Protease from Aspergillus oryzae by Introduction of a Disulfide Bond. Bioscience, Biotechnology, and Biochemistry: Vol. 56, No. 2, pp. 326-327.
- Published
- 1992
37. A novel selective peroxisome proliferator-activated receptor alpha agonist, 2-methyl-c-5-[4-[5-methyl-2-(4-methylphenyl)-4-oxazolyl]butyl]-1,3-dioxane-r-2-carboxylic acid (NS-220), potently decreases plasma triglyceride and glucose levels and modifies lipoprotein profiles in KK-Ay mice.
- Author
-
Kenji, Kuwabara, Kohji, Murakami, Makoto, Todo, Tomiyoshi, Aoki, Tetsuo, Asaki, Masatoshi, Murai, and Junichi, Yano
- Abstract
2-Methyl-c-5-[4-[5-methyl-2-(4-methylphenyl)-4-oxazolyl]butyl]-1,3-dioxane-r-2-carboxylic acid (NS-220) was newly synthesized and demonstrated to be a novel potent peroxisome proliferator-activated receptor alpha (PPARalpha) agonist with high subtype selectivity. In cell-based reporter gene assays, the EC(50) values of NS-220 for human PPARalpha, PPARgamma, and PPARdelta were 1.9 x 10(-8), 9.6 x 10(-6), and >10(-4) M, respectively, and for mouse PPARalpha, PPARgamma, and PPARdelta were 5.5 x 10(-8), 3.3 x 10(-5), and >10(-4) M, respectively. In addition, [(3)H]NS-220 bound to the ligand-binding domain of human PPARalpha with a K(D) value of 1.85 x 10(-7) M. Fenofibric acid and bezafibrate showed weak agonist activity for PPARalpha (EC(50), 2-8 x 10(-5) M), with poor subtype selectivity. NS-220 (0.1-3 mg/kg p.o.) decreased plasma triglyceride levels in ddY mice in a dose-dependent manner, but its hypolipidemic activity was abolished in PPARalpha-deficient mice. In KK-A(y) mice, an animal model of type-2 diabetes, NS-220 (0.3-1 mg/kg p.o.; 4 days) and fenofibrate (100-300 mg/kg p.o.; 4 days) decreased plasma triglyceride and glucose levels in a dose-dependent manner. In a 2-week repeated administration test, NS-220 (0.3-1 mg/kg p.o.) decreased plasma glucose levels markedly without increasing in plasma insulin levels. Furthermore, NS-220 increased high-density lipoprotein levels and decreased triglyceride-rich lipoprotein levels. In conclusion, a newly synthesized dioxanecarboxylic acid derivative, NS-220, is a potent and highly selective PPARalpha agonist that ameliorates metabolic disorders in diabetic mice. These results strongly suggest that it will be a promising drug for the treatment of hyperlipidemia or metabolic disorders in type-2 diabetes.
- Published
- 2004
38. Renal cell carcinoma with intracaval tumor thrombus extending to the diaphragm: ultrasonography and surgical management
- Author
-
Shohzoh Nakagawa, Masanobu Manabe, Takayasu Yoh, Norihide Taniuchi, Hideki Komatsu, Mario Miyamoto, and Kohji Murakami
- Subjects
Male ,medicine.medical_specialty ,Urology ,Celiac axis ,Vena Cava, Inferior ,Inferior vena cava ,Tumor thrombus ,Renal cell carcinoma ,medicine.artery ,medicine ,Humans ,cardiovascular diseases ,Thrombus ,Carcinoma, Renal Cell ,Aged ,Ultrasonography ,Aorta ,business.industry ,medicine.disease ,Kidney Neoplasms ,Diaphragm (structural system) ,Surgery ,medicine.vein ,cardiovascular system ,Radiology ,business ,Tomography, X-Ray Computed - Abstract
We describe the successful management of a patient with intracaval extension of renal cell carcinoma to the level of the diaphragm. Ultrasonography delineated clearly the upper extent of the tumor thrombus and revealed no invasion of the vena caval wall at or above the entrance of the main hepatic veins. To prevent severe hypotension caused by intrapericardial control of the inferior vena cava, the aorta was clamped temporarily just above the celiac axis. The tumor was removed completely en bloc with the thrombus. Blood loss was minimal and the patient recovered promptly.
- Published
- 1985
39. A full length cDNA clone for the alkaline protease from Aspergillus oryzae: structural analysis and expression in Saccharomyces cerevisiae
- Author
-
Yutaka Ishida, Yoshihiro Ogawa, Hiroki Tatsumi, Haruhide Kawabe, Hiroshi Motai, Eiichi Nakano, Atsushi Masaki, Hirofumi Arimura, Seiji Murakami, and Kohji Murakami
- Subjects
Aspergillus oryzae ,Blotting, Western ,Molecular Sequence Data ,Restriction Mapping ,Gene Expression ,Saccharomyces cerevisiae ,Biology ,Complementary DNA ,Sequence Homology, Nucleic Acid ,Genetics ,Consensus sequence ,Amino Acid Sequence ,Subtilisins ,Cloning, Molecular ,DNA, Fungal ,Molecular Biology ,Peptide sequence ,chemistry.chemical_classification ,Enzyme Precursors ,Base Sequence ,Serine Endopeptidases ,Protein primary structure ,Subtilisin ,Nucleic acid sequence ,biology.organism_classification ,Molecular biology ,Amino acid ,Aspergillus ,chemistry ,Biochemistry ,Plasmids - Abstract
We have cloned and determined the nucleotide sequence of a cDNA fragment for the entire coding region of the alkaline protease (Alp) from a filamentous ascomycete Aspergillus oryzae. According to the deduced amino acid sequence, Alp has a putative prepro region of 121 amino acids preceding the mature region, which consists of 282 amino acids. A consensus sequence of a signal peptide consisting of 21 amino acids is found at the N-terminus of the prepro region. The primary structure of the mature region shares extensive homology (29%-44%) with those of subtilisin families, and the three residues (Asp 32, His 64 and Ser 221 in subtilisin BPN') composing the active site are preserved. The entire cDNA, coding for prepro Alp, when introduced into the yeast Saccharomyces cerevisiae, directed the secretion of enzymatically active Alp into the culture medium, with its N-terminus and specific activity identical to native Aspergillus Alp.
- Published
- 1989
40. Cloning and sequencing of the alkaline protease cDNA from Aspergillus oryzae
- Author
-
Haruhide Kawabe, Kohji Murakami, Yutaka Ishida, Atsushi Masaki, Eiichi Nakano, Hiroshi Motai, Hirofumi Arimura, Hiroki Tatsumi, Manabu Ohsawa, Ryohei Tsuji, and Seiji Murakami
- Subjects
Cloning ,chemistry.chemical_classification ,biology ,Chemistry ,Nucleic acid sequence ,Fungi imperfecti ,Molecular cloning ,biology.organism_classification ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Enzyme ,Aspergillus oryzae ,Biochemistry ,Complementary DNA ,General Agricultural and Biological Sciences ,Gene - Published
- 1988
41. Cloning and Sequencing of the Alkaline Protease cDNA fromAspergillus oryzae
- Author
-
Hiroki Tatsumi, Manabu Ohsawa, Ryohei F. Tsuji, Seiji Murakami, Eiichi Nakano, Hiroshi Motai, Atsushi Masaki, Yutaka Ishida, Kohji Murakami, Haruhide Kawabe, and Hirofumi Arimura
- Subjects
General Agricultural and Biological Sciences ,General Biochemistry, Genetics and Molecular Biology - Published
- 1988
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