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Biomedical subjects

M Kubota

Publications and source records attributed to M Kubota.

At least 73 records · Page 4Linked to original sources
PubMed2001

[Analysis of inquiries about prescriptions at the pharmacy of Gifu Pharmaceutical University].

The Ministry of Health, Labour and Welfare explains the objectives of promotion of separation of the pharmacy and clinics as the elimination of duplicated prescription of similar drugs and drug interactions caused by treatment at several departments or hospitals, and sufficient guidance in the use of drugs by pharmacists. The Pharmacy of Gifu Pharmaceutical University has dispensed prescriptions by outside medical organizations as its routine activity. In this study, the contents of question inquiries handled in routine activities were compiled and analyzed, and their meaning was evaluated. The contents of question inquiries were accumulated using data cards. The data obtained during 2 and a half years (562 cases) were analyzed. The percentage of the number of inquiries relative to the number of prescriptions was highest at 1.86% during the first 3 months and was 1.05-1.71% per 3 months thereafter. The inquiries were most frequently about "the dosage/regimen" (153 cases), followed by "discrepancy between the contents of prescription and understanding of the patient" (88 cases) and "problems about insurance coverage" (80 cases). There were also 16 inquiries about "the possibility of contraindications and adverse reactions" and 15 inquiries about "duplicated prescription", which may have exerted serious effects on the patients. Eighty nine % of the inquiries have led to changes in the prescriptions, and about half of these cases were discovered by consultation with the patients or a review of the drug history. Proper and positive execution of these operations in routine pharmacy work is considered to lead to appropriate inquiries about problems with prescriptions and, thus, contribute to the proper use of drugs and prevention of malpractice.

Drug Administration Schedule↗
PubMed2001

Trehalose-producing operon treYZ from Arthrobacter ramosus S34.

Arthrobacter ramosus S34, which produces trehalose from maltooligosaccharide, was isolated. A trehalose-producing operon, treYZ, was cloned from the genome. Expression experiments with treY and treZ confirmed that they coded malto-oligosyltrehalose synthase and malto-oligosyltrehalose trehalohydrolase, respectively. The amino acid sequence of TreY from A. ramosus S34 and that from Arthrobacter sp. Q36 did not show high identity, nor did those of TreZ.

Amino Acid Sequence↗
PubMed2001

Purification and substrate specificity of honeybee, Apis mellifera L., alpha-glucosidase III.

Alpha-glucosidase III, which was different in substrate specificity from honeybee alpha-glucosidases I and II, was purified as an electrophoretically homogeneous protein from honeybees, by salting-out chromatography, DEAE-cellulose, DEAE-Sepharose CL-6B, Bio-Gel P-150, and CM-Toyopearl 650M column chromatographies. The enzyme preparation was confirmed to be a monomeric protein and a glycoprotein containing about 7.4% of carbohydrate. The molecular weight was estimated to approximately 68,000, and the optimum pH was 5.5. The substrate specificity of alpha-glucosidase III was kinetically investigated. The enzyme did not show unusual kinetics, such as the allosteric behaviors observed in alpha-glucosidases I and II, which are monomeric proteins. The enzyme was characterized by the ability to rapidly hydrolyze sucrose, phenyl alpha-glucoside, maltose, and maltotriose, and by extremely high Km for substrates, compared with those of alpha-glucosidases I and II. Especially, maltotriose was hydrolyzed over 3 times as rapidly as maltose. However, maltooligosaccharides of four or more in the degree of polymerization were slowly degraded. The relative rates of the k0 values for maltose, sucrose, p-nitrophenyl alpha-glucoside and maltotriose were estimated to be 100, 527, 281 and 364, and the Km values for these substrates, 11, 30, 13, and 10 mM, respectively. The subsite affinities (Ai's) in the active site were tentatively evaluated from the rate parameters for maltooligosaccharides. In this enzyme, it was peculiar that the Ai value at subsite 3 was larger than that of subsite 1.

Animals↗
PubMed2001

Crystallization and structural analysis of intact maltotetraose-forming exo-amylase from Pseudomonas stutzeri.

The intact maltotetraose-forming exo-amylase from Pseudomonas stutzeri (G4-1), which has a raw starch binding domain, has been crystallized. The structure was identified (PDB entry 1GCY) by the molecular replacement method using the structure of its catalytic domain (G4-2). The result showed that the raw starch binding domain is in a disordered state, the corresponding electron densities being almost invisible. Superposition of these two enzyme forms showed evidence for the possible location of the raw starch binding domain (SBD). This crystal is a novel case, in that it forms a regular lattice incorporating flexibly bound SBD in the channel of crystal packing of the catalytic domains.

Binding Sites↗
PubMed2001

The polymorphism in the caudal-related homeodomain protein Cdx-2 binding element in the human vitamin D receptor gene.

The major physiological activity of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is the regulation of calcium absorption in the small intestine, and the level of vitamin D receptor (VDR) is an important factor in this regulation. In a previous study, we indicated-that the caudal-related homeodomain Cdx-2 played an important role in the intestine-specific transcription of the human VDR gene. In this study, the polymorphism was identified in the core sequence 5'-ATAAAAACTTAT-3' in the Cdx-2 binding site in the VDR gene promoter. In 261 Japanese women with genotyped VDR polymorphisms, 48 were genotype Cdx-A (adenine at -3731 nucleotides [nt] relative to the transcription start site of human VDR gene 5-ATAAAAACTTAT), 82 were genotype Cdx-G (guanine at -3731 nt, 5'-GTAAAAACTTAT-3'), and 131 were genotype Cdx-A/G (heterozygote). In postmenopausal Japanese women, the bone mineral density (BMD) in the lumbar spine (L2-L4) with the Cdx-G homozygote was 12% lower than that with the Cdx-A homozygote (p < 0.05). In electrophoretic gel mobility shift assay (EMSA), the oligonucleotide with Cdx-G allele markedly decreased the binding to Cdx-2 compared with that in the Cdx-A allele. The transcriptional activity of the VDR promoter with Cdx-G allele was decreased to 70% of the Cdx-A allele. In addition, in the herpes simplex virus thymidine kinase promoter, the Cdx-2 binding element with the G allele showed significantly lower transcriptional activity than that of the A allele. Thus, the polymorphism in the Cdx-2 binding site of the VDR gene (Cdx-polymorphism) would affect the expression of VDR in the small intestine. In addition, this polymorphism may modulate BMD in postmenopausal Japanese women.

Adult↗
PubMed2001

Infarction followed by hemorrhage in pituitary adenoma due to endocrine stimulation test.

A 63-year-old man, who presented with visual field loss due to pituitary tumor, received an intravenous bolus injection of thyrotropin and gonadotropin releasing hormones and insulin as a preoperative evaluation. He complained of severe headache and nausea 2 hours after injection. Emergent CT scan showed no evidence of intratumoral hemorrhage. The next day, his visual field became null. MR images revealed heterogeneous mixed intensity lesions. Under diagnosis of pituitary apoplexy, he underwent transsphenoidal tumor removal 30 hours after onset. Intraoperative and pathological findings showed tumor hemorrhage and adjacent necrotic change. Fourteen cases with sufficient clinical detail in the literature are reviewed: All of the cases had macroadenoma with suprasellar extension. Testing agents were gonadotropin and thyrotropin releasing hormones in 92.9% and 85.7% of cases, respectively. Headache was an initial symptom and started within two hours in all cases but one. Half of the cases showed no change on CT scan. However, tumor hemorrhage was evidenced in 92.9% of cases with or without necrosis due to ischemic change, intraoperatively or pathologically. It is speculated that pituitary apoplexy often starts with infarction possibly due to vasoactive effect of testing agents and later develops into hemorrhage. Therefore, it is necessary to observe patients closely at least a few hours after endocrine stimulation test, and MR imaging may make an earlier diagnosis for the pituitary apoplexy since CT scan often shows no density change in the pituitary adenoma.

Adenoma↗
PubMed2001

Somatic-cell mutation induced by UVA and monochromatic UV radiation in repair-proficient and -deficient Drosophila melanogaster.

Near-ultraviolet light (UVA: 320-400 nm) constitutes a major part of sunlight UV. It is important to know the effect of UVA on the biological activities of organisms on the earth. We have previously reported that black light induces somatic-cell mutation in Drosophila larvae. To investigate which wavelength of the UVA is responsible for the mutation we have now carried out a series of monochromatic irradiations (310, 320, 330, 340, 360, 380 and 400 nm) on Drosophila larvae, using the large spectrograph of the National Institute for Basic Biology (Okazaki National Research Institutes, Okazaki, Japan). Mutagenic activity was examined by the Drosophila wing-spot test in which we observe mutant wing hair colonies (spots) on the wings of adult flies obtained from the treated larvae. The induction of mutation was highest by irradiation at 310 nm and decreased as the wavelength became longer. Neither the 380 nor the 400 nm light was mutagenic. Excision repair is known to protect cells from UV damage. In the excision-repair-deficient Drosophila, the mutagenic response induced by 310 nm irradiation was 24-fold higher than that of the wild-type (7.2 +/- 1.5 spots/wing/kJ vs 0.3 +/- 0.08 spots/wing/kJ), and at 320 nm the difference of the response was 14-fold (0.21 vs 0.015 +/- 0.005). In the case of irradiation at 330 and 340 nm the difference of the response was only two-fold (at 330 nm, 6.9 +/- 2.9 x 10(-3) vs 3.1 +/- 1.1 x 10(-3) spots/wing/kJ; at 340 nm, 3.5 +/- 0.9 x 10(-3) vs 2.0 +/- 0.7 x 10(-3). These results suggest that the lesion caused in the larvae by 320 nm irradiation may be similar to the damage induced by 310 nm and that the lights of 330 and 340 nm may induce damage different from the lesions induced by shorter-wavelength lights.

Animals↗
PubMed2001

[Childhood t(8;21) acute myelocytic leukemia: a comparison of clinical features and risk factors with adult cases].

Despite the abundance of reports describing adult cases of t(8;21) acute myelocytic leukemia (AML), childhood cases have received little attention. We retrospectively investigated 14 childhood cases of t(8;21) AML, and compared their clinical characteristics with those of adult cases, focusing on the risk factors for poor prognosis. Seventy-one percent of the patients had fever. Their mean leukocyte count was 12,700/microliter, and they showed decreased NAP activity. The cell surface showed positivity for CD13, 33, 19, 34, and HLA-DR. The complete remission rate was 100%, and relapse was observed in three of the patients. Bone marrow eosinophilia was present in a smaller proportion of the childhood cases than in the adult cases. Although an increased leukocyte count, tumor formation, and other risk factors have been reported in adults, there was no correlation between these factors and prognosis in our childhood cases. As children who showed AML relapse had TdT-positive blasts, detectable blast TdT activity may be a risk factor for relapse in childhood cases of t(8;21) AML. However, to confirm this, a study with a larger subject base should be conducted.

Adolescent↗
PubMed2001

A scoring system for ultrasonographic differentiation between cervical malignant lymphoma and benign lymphadenitis.

The purpose of our study is to assess the usefulness of a scoring system for ultrasonography using a high-frequency transducer as a non-invasive examination in the differential diagnosis of cervical malignant lymphoma from benign lymphadenitis. Ultrasonography was performed with a real-time linear scanner using 7.5 and/or 11 MHz probes in 66 patients who were presented with cervical lymphadenopathy and were suspected to have malignant lymphoma. Thereafter, a surgical biopsy was performed for histopathological diagnosis, revealing 49 cases of malignant lymphoma and 17 cases of benign lymphadenitis. Ultrasonographic findings were reviewed retrospectively. The incidence of following ultrasonographic features was found to be significantly higher in malignant lymphoma than in benign lymphadenitis. These were; (1) longitudinal and transverse diameter ratio(L/T) < 2.0, (2) a delineation of multiple nodes, (3) a tendency of fusion, (4) an irregular margin, (5) hypoechoic masses with heterogeneous internal echoes, (6) a presence of line echoes and (7) an absence of hilum. These 7 ultrasonographic features were used in combination for a scoring system to discriminate malignant lymphoma from benign lymphadenitis. When ultrasonographic findings of lymph nodes fulfilled more than 3 of the 7 features, malignant lymphoma was diagnosed with a sensitivity of 98.0% and a specificity of 76.5%. Ultrasonographic evaluation of swollen cervical lymph nodes using a high-frequency transducer would be informative and should be performed prior to a surgical biopsy.

Adolescent↗
PubMed2001

Heavily T2-weighted MR imaging of white matter tracts in the hypothalamus: normal and pathologic demonstrations.

BACKGROUND AND PURPOSE: The MR appearance of white matter tracts in the hypothalamus and the role of the hypothalamus as a memory mechanism have not been sufficiently described in clinical settings. Heavily T2-weighted black-and-white reversed (T2R) images were assessed to reveal their visualization and clinical significance. METHODS: One hundred healthy subjects and three patients with hypothalamic lesions underwent fast spin-echo MR imaging to reveal the postcommissural fornix (PF) and mammillothalamic tract (MT). RESULTS: The PF was identifiable in axial and/or coronal sections in all healthy subjects. No remarkable asymmetry of its size or course was evident. Both anteroposterior and vertical dimensions ranged from 10.5 to 14 mm. The MT was visible in one or two axial sections above the mammillary body in 64% of healthy subjects and in a coronal section in 36%. Two patients with glioblastoma multiforme and lacunar infarct at the hypothalamus presented with anterograde amnesia; T2R imaging revealed involvement of both the PF and MT. The third patient had a suprasellar craniopharyngioma with PF injury sparing the MT resulting from surgical manipulation and was free of memory deficit. Anterograde amnesia was evident only when both the PF and MT were injured. CONCLUSION: T2R images have made a high rate of detection of the PF and MT possible and could provide a more detailed correlation of hypothalamic neuroanatomy and memory mechanism in clinical settings.

Adolescent↗
PubMed2001

[Hematopoietic stem cell transplantation with busulfanthiotepa-cyclophosphamide conditioning for pediatric patients with high-risk acute lymphoblastic leukemia].

Twelve children with high-risk acute lymphoblastic leukemia underwent stem cell transplantation (SCT) with a conditioning regimen consisting of busulfan, cyclophosphamide and thiotepa. Eight of them underwent SCT while in complete remission (CR) and the other 4 while not in CR. Three children underwent HLA-matched related bone marrow transplantation (BMT), 7 HLA-matched unrelated BMT, 1 HLA one-locus-mismatched unrelated cord blood cell transplantation, and 1 autologous peripheral blood stem cell transplantation. Grade II-IV acute GVHD was observed in 3 of the 11 allo-SCT cases, while chronic GVHD was seen in 3 of 9 evaluable cases. None of the 12 cases showed thrombotic microangiopathy, and veno-occlusive disease (VOD) was observed in 3. Nine of the patients are alive and disease-free 6-45 months after diagnosis. The event-free survival rate at 3 years was 72.2% for the 12 patients, including 8 of the 9 who received SCT during CR, and 2 of the 4 who did so while not in CR. The other 3 patients died: 2 of disease progression and 1 of VOD with pneumonia. All of those who died had undergone unrelated BMT.

Adolescent↗
PubMed2001

Implantation of presternal catheter using Moncrief technique: aiming for fewer catheter-related complications.

OBJECTIVES: To reduce catheter-related complications, we developed a new technique of catheter implantation, combining a presternal catheter with the Moncrief technique. METHODS: The presternal catheter, consisting of 2 catheters joined by a titanium extender, was surgically implanted. Its end was left embedded in the presternal wall. A few weeks after implantation, the embedded subcutaneous catheter was exteriorized, exiting in the 4th intercostal space, and peritoneal dialysis (PD) was commenced. RESULTS: Using the new technique, 9 catheters were implanted (3 in women and 6 in men). Exteriorization was performed 30.6 +/- 14.3 days after implantation of the catheter. Total observation period was 70 patient-months. Average hospitalization was 4.4 +/- 1.3 days for catheter implantation, and 2.6 +/- 2.6 days for exteriorization. Peritoneal dialysis commenced on the day of exteriorization with an exchange volume of 1.8 +/- 0.3 L, using 4 exchanges daily. During the observation period, none of the patients experienced a catheter infection or dialysate leak. One non infectious complication was observed (a catheter wrapped in omentum). CONCLUSIONS: Our approach of combining a presternal catheter and the Moncrief technique had some advantages not only in regard to catheter infection and dialysate leakage, but also in regard to quality of life and hospitalization for the patient.

Adult↗
PubMed2000

Association between two types of vitamin d receptor gene polymorphism and bone status in premenopausal Japanese women.

Polymorphisms in the vitamin D receptor (VDR) gene using ultrasound (US) bone mass and bone metabolic markers were investigated as potential genetic markers for osteoporosis in 126 premenopausal Japanese women aged 27.2 +/- 10.1 (mean +/- SD) years. The relationship between their VDR gene polymorphisms and bone states was determined. VDR genotypes were based on the absence (B) or presence (b) of the Bsm I restriction site (B polymorphism), and ATG (the M allele) and ACG (the m allele) sequences at the translation initiation site (M polymorphism). Genotype frequencies were 73.8%, bb; 24.6%, Bb; 1.6%, BB; 15.1%, MM; 51.6%, Mm and 33.3%, mm. The stiffness index of calcaneal bone minerals measured by an US bone densitometer was significantly higher in the mm types (P <0.05 versus MM) than in the Mm types (P <0.01 versus MM) and MM types. There was no significant difference between in B polymorphisms. Furthermore, bone mass was correlated with serum bone type alkaline phosphatase (ALP) activity and urinary deoxypyridinoline concentration in M polymorphisms. Because the distribution of B polymorphisms in each M polymorphism genotype did not differ, M polymorphisms were affected independently from B polymorphisms to bone mass or bone metabolic markers. No significant difference was observed in nutritional intake and food consumption among genotypes. In the MM and Mm types, the bone mass was closely related to the frequency of milk intake during the periods of elementary and junior high school. In contrast, bone mass was not associated with nutritional intake or the frequency of past milk intake in B polymorphisms. Therefore, the M polymorphism of the VDR gene is a stronger genetic indicator of osteoporosis than the B polymorphism in premenopausal Japanese woman.

Adult↗
PubMed2000

Lethal forebrain ischemia stimulates sphingomyelin hydrolysis and ceramide generation in the gerbil hippocampus.

Ceramide, a hydrolyzed product of sphingomyelin, is reported to play an important role in apoptosis. In this study, we measured the sphingomyelin and ceramide levels in the hippocampus of the gerbil after transient forebrain ischemia for 5 min (lethal) or 2 min (sublethal). The aim was to examine alterations in the sphingomyelin cycle during delayed neuronal death, which we considered could be due to apoptosis. Sphingolipids were separated on high-performance thin-layer chromatography plates and analyzed by gas-liquid chromatography. At 30 min and 24 h after lethal ischemia, sphingomyelin levels were decreased and ceramide levels were increased compared with control levels. No significant changes were observed after sublethal ischemia. These results suggest that the sphingomyelin cycle may have a role in neuronal death.

Animals↗
PubMed2000

Crystal structure of NADH-dependent ferredoxin reductase component in biphenyl dioxygenase.

Oxidative biodegradation of aromatic compounds by bacteria usually begins with hydroxylation of the aromatic ring by multi-component dioxygenases like benzene dioxygenase, biphenyl dioxygenase, and others. These enzymes are composed of ferredoxin reductase, ferredoxin, and terminal oxygenase. Reducing equivalents that originate from NADH are transferred from ferredoxin reductase to ferredoxin and, in turn, to the terminal oxygenase, thus resulting in the activation of a dioxygen. BphA4 is the ferredoxin reductase component of biphenyl dioxygenase from Pseudomonas sp. strain KKS102. The amino acid sequence of BphA4 exhibits significant homology with the putidaredoxin reductase of the cytochrome P450cam system in Pseudomonas putida, as well as with various other oxygenase-coupled NADH-dependent ferredoxin reductases (ONFRs) of bacteria. To date, no structural information has been provided for the ferredoxin reductase component of the dioxygenase systems. In order to provide a structural basis for discussing the mechanism of electron transport between ferredoxin reductase and ferredoxin, crystal structures of BphA4 and its NADH complex were solved. The three-dimensional structure of BphA4 is different from those of ferredoxin reductases whose structures have already been determined, but adopts essentially the same fold as the enzymes of the glutathione reductase (GR) family. Also the three-dimensional structure of the first two domains of BphA4 adopts a fold similar to that of adrenodoxin reductase (AdR) in the mitochondrial cytochrome P450 system. Comparing the amino acid sequence with what is known of the three-dimensional structure of BphA4 strongly suggests that the other ONFRs have secondary structural features that are similar to that of BphA4. This analysis of the crystal structures of BphA4 suggests that Lys53 and Glu159 seem to be involved in the hydride transfer from NADH to FAD. Since the amino acid residues around the active site, some of which seem to be important to electron transport, are highly conserved among ONFRs, it is likely that the mechanism of electron transport of BphA4 is quite applicable to other ONFRs.

Amino Acid Sequence↗
PubMed2000

Visualization, quantification and therapeutic evaluation of angiogenic vessels in cancer by synchrotron microangiography.

The usefulness of a synchrotron microangiography system for depicting, quantitating and therapeutically evaluating angiogenic vessels in cancer is illustrated. In 20 mice transplanted with murine colon cancer, sequential changes in the angiogenic vessels were determined by using synchrotron microangiography, using changes in tumor volume for reference. This system allowed the depiction and quantification of angiogenic vessels in the period from one to four weeks after transplantation. The effects of antiangiogenic therapy were evaluated by using a neutralizing antibody against vascular endothelial growth factor. The neutralizing antibody partially suppressed angiogenesis and tumor growth. Synchrotron microangiography is shown to be useful for the depiction, quantification and evaluation of angiogenic vessels in cancer.

Journal Article↗