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Biomedical subjects

Y Nagata

Publications and source records attributed to Y Nagata.

At least 145 records · Page 8Linked to original sources
PubMed1999

Identification of the catalytic triad in the haloalkane dehalogenase from Sphingomonas paucimobilis UT26.

The haloalkane dehalogenase from Sphingomonas paucimobilis UT26 (LinB) is the enzyme involved in the gamma-hexachlorocyclohexane degradation. This enzyme hydrolyses a broad range of halogenated aliphatic compounds via an alkyl-enzyme intermediate. LinB is believed to belong to the family of alpha/beta-hydrolases which employ a catalytic triad, i.e. nucleophile-histidine-acid, during the catalytic reaction. The position of the catalytic triad within the sequence of LinB was probed by a site-directed mutagenesis. The catalytic triad residues of the haloalkane dehalogenase LinB are proposed to be D108, H272 and E132. The topological location of the catalytic acid (E132) is after the beta-strand six which corresponds to the location of catalytic acid in the pancreatic lipase, but not in the haloalkane dehalogenase of Xanthobacter autotrophicus GJ10 which contains the catalytic acid after the beta-strand seven.

Binding Sites↗
PubMed1999

D-Aspartate stimulation of testosterone synthesis in rat Leydig cells.

D-Aspartate increases human chorionic gonadotropin-induced testosterone production in purified rat Leydig cells. L-Aspartate, D-,L-glutamate or D-,L-asparagine could not substitute for D-aspartate and this effect was independent of glutamate receptor activation. Testosterone production was enhanced only in cells cultured with D-aspartate for more than 3 h. The increased production of testosterone was well correlated with the amounts of D-aspartate incorporated into the Leydig cells, and L-cysteine sulfinic acid, an inhibitor of D-aspartate uptake, suppressed both testosterone production and intracellular D-aspartate levels. D-Aspartate therefore is presumably taken up into cells to increase steroidogenesis. Intracellular D-aspartate probably acts on cholesterol translocation into the inner mitochondrial membrane, the rate-limiting process in steroidogenesis.

Animals↗
PubMed1999

A longitudinal study of the effect of vitamin K2 on bone mineral density in postmenopausal women a comparative study with vitamin D3 and estrogen-progestin therapy.

OBJECTIVES: To investigate the effect of vitamin K2 treatment for a year on spinal bone mineral density (BMD) in postmenopausal women, comparing with vitamin D3 hormone replacement therapy and to determine the factors which affect the efficacy of vitamin K2 therapy. SUBJECTS AND METHODS: Seventy-two postmenopausal women were randomized into four groups and treated with respective agents. Before the therapy, 6 and 12 months after the treatment, their lumbar spine BMD were measured by dual energy X-ray absorptiometry. The rates of change in BMD (delta BMD) were calculated. Correlations of BMD with age, year since menopause and the initial BMD were determined. RESULTS: Vitamin K2 suppressed the decrease in spinal BMD as compared with no treatment group. BMD in women treated with vitamin K2 was inversely correlated with their age (r = -0.54; P < 0.05). CONCLUSIONS: Vitamin K2 therapy may be a useful method for preventing postmenopausal spinal bone mineral loss. In addition, the therapy should be started early in postmenopausal period.

Bone Density↗
PubMed1999

Bone mineral density in postmenopausal women with endometrial cancer.

OBJECTIVE: The aim of the study was to investigate the relationship between endometrial cancer and bone mineral density (BMD). MATERIALS AND METHODS: A total of 55 postmenopausal Japanese women with well-differentiated adenocarcinoma and 284 age-matched healthy women were studied. Baseline characteristics including age, age at menopause, years since menopause (YSM), weight, height, body mass index (BMI), prior menstrual history, parity, and fertility were recorded for each subject. Lumbar spine BMD (L2-4), and body fat indices including body fat mass amount and percent body fat were measured by dual-energy X-ray absorptiometry. These variables were compared between the two groups. In all subjects (n = 339), correlations of BMD with the presence of endometrial cancer, baseline characteristics, and body fat indices were investigated, using univariate and multiple regression analyses. RESULTS: BMD, weight, BMI, body fat mass amount, and percent body fat were significantly higher in women with endometrial cancer. Other baseline characteristics did not differ between the two groups. Age at menopause, height, weight, BMI, percent body fat, and body fat mass amount were positively correlated with BMD, while age and YSM were inversely correlated with BMD. After adjusting for age, YSM, and height, the presence of endometrial cancer was still correlated with BMD (P < 0.05). However, after adding body fat mass amount to these three adjusted variables, there was no correlation between the presence of endometrial cancer and BMD. CONCLUSION: Women with endometrial cancer have a high BMD. This is attributable to high body fat mass amount in this disease.

Adenocarcinoma↗
PubMed1999

Construction and characterization of histidine-tagged haloalkane dehalogenase (LinB) of a new substrate class from a gamma-hexachlorocyclohexane-degrading bacterium, Sphingomonas paucimobilis UT26.

The linB gene product (LinB), which is involved in the degradation of gamma-hexachlorocyclohexane in Sphingomonas paucimobilis UT26, is a member of haloalkane dehalogenases with a broad range of substrate specificity. Elucidation of the factors determining its substrate specificity is of interest. Aiming to facilitate purification of recombinant LinB protein for site-directed mutagenesis analysis, a 6-histidyl tail was added to the C-terminus of LinB. The His-tagged LinB was specifically bound with Ni-NTA resin in the buffer containing 10 mM imidazole. After elution with 500 mM imidazole, quantitative recovery of protein occurred. The steady-state kinetic parameters of the His-tagged LinB for four substrates were in good agreement with that of wild-type recombinant LinB. Although the His-tagged LinB expressed in an average of 80% of the activity of the wild type LinB for 10 different substrates, the decrease was very similar for different substrates with the standard deviation of 5.5%. The small activity reduction is independent of the substrate shape, size, or number of substituents, indicating that the His-tagged LinB can be used for further mutagenesis studies. To confirm the suitability of this system for mutagenesis studies, two mutant proteins with substitution in putative halide binding residues (W109 and F151) were constructed, purified, and tested for activity. As expected, complete loss in activity of W109L and sustained activity of F151W were observed.

Base Sequence↗
PubMed1999

A double-masked multicenter comparative study between alendronate and alfacalcidol in Japanese patients with osteoporosis. The Alendronate Phase III Osteoporosis Treatment Research Group.

To evaluate the efficacy and safety of alendronate, a double-masked, active (alfacalcidol) controlled comparative study for 48 weeks was carried out in a total of 210 Japanese patients with osteoporosis. The doses of alendronate and alfacalcidol were 5 mg/day and 1 microgram/day, respectively. The lumbar bone mineral density (LBMD) values observed at 12, 24, 36 and 48 weeks after the initiation of alendronate treatment were 3.53 +/- 0.53%, 5.37 +/- 0.62%, 5.87 +/- 0.74% and 6.21 +/- 0.59% (mean +/- SE), respectively, higher than the baseline value. Corresponding values in the alfacalcidol group were 1.50 +/- 0.43%, 0.69 +/- 0.63%, 1.12 +/- 0.60% and 1.36 +/- 0. 63%, respectively. There was a significant difference between the two groups at each time point (p<0.05 or p<0.001). The bone turnover markers were depressed during treatment in the alendronate group: -32.2% for alkaline phosphatase, -53.7% for N-terminal osteocalcin and -45.0% for urinary deoxypyridinoline compared with the corresponding baseline values. On the contrary, no notable changes in these parameters were observed in the alfacalcidol group. Treatment with alendronate caused a transient decrease in serum calcium concentrations associated with an increase in the serum level of intact parathyroid hormone. In contrast, treatment with alfacalcidol resulted in a tendency of these parameters to change in the opposite direction. No difference in fracture incidence between the two groups was observed. The overall safety of alendronate was comparable to that of alfacalcidol. In conclusion, although it was a relatively short-term study of 48 weeks, the results of the present study indicate that alendronate at the daily dose of 5 mg was effective in increasing LBMD and that no serious drug-related adverse events were observed in the alendronate-treated patients. Alendronate is more efficacious than alfacalcidol in increasing bone mineral density, although the mechanisms of the actions of the two drugs are apparently different.

Aged↗
PubMed1999

Measurement of obstetric conjugate by ultrasonic tomography and its significance.

OBJECTIVE: The purpose of this study was to investigate the clinical usefulness of the measurement of ultrasonic obstetric conjugate. STUDY DESIGN: In 209 pregnant women, ultrasonic obstetric conjugate was measured twice at 28 and 36 weeks of pregnancy. In 26 of these patients the measurement of obstetric conjugate by x-ray pelvimetry was also performed at 36 weeks of pregnancy because of medical indications. Obstetric outcome was investigated in association with ultrasonic obstetric conjugate. The relationship between ultrasonic obstetric conjugate and x-ray pelvimetry was investigated. RESULTS: Ultrasonic obstetric conjugate ranged from 10.7 to 15.1 cm. In 12 patients (5.7%) ultrasonic obstetric conjugate was less than 12 cm. Six of them (50.0%) underwent cesarean section because of dystocia. This percentage was significantly higher than 7.1% of the cesarean section rate (14 of 197) in patients with ultrasonic obstetric conjugate more than 12 cm (P <.001). A close positive correlation was observed between ultrasonic obstetric conjugate and x-ray pelvimetry (r = 0.91; P <.0001). CONCLUSIONS: We conclude that ultrasonic obstetric conjugate measurement is a safe and useful procedure in the prediction of dystocia.

Adult↗
PubMed1999

Pharmacokinetics of exogenous gonadotropin and ovarian response in in vitro fertilization.

OBJECTIVE: To assess the diffusion of gonadotropin into the follicular fluid (FF) and its relation to the results achieved in a human IVF-ET program. DESIGN: Retrospective pharmacokinetic study. SETTING: Fukuoka University Hospital, Japan. PATIENT(S): Eighty-seven infertile patients underwent 137 cycles of IVF-ET. INTERVENTION(S): Serum and FF were collected at the time of oocyte recovery. The hCG ratio (between follicular hCG and serum hCG concentrations, measured by time-resolved fluoroimmunoassay) was evaluated as an index of the diffusion of exogenous gonadotropin. MAIN OUTCOME MEASURE(S): Relation between hCG ratio and the results and outcome of the IVF-ET program. RESULT(S): The hCG ratio decreased with the total dosage of hMG and increased with the serum E2 level, the number of oocytes recovered, and the number of oocytes fertilized. Patients with a poor response showed a low hCG ratio, which was associated with a complete lack of fertilization. The mean hCG ratio in the pregnant cycles was significantly higher than that in the nonpregnant cycles. An hCG ratio > 0.46 was seen in all pregnant cycles. CONCLUSION(S): The diffusion of exogenous gonadotropin into the FF may be an important predictor of IVF outcome.

Chorionic Gonadotropin↗
PubMed1999

Serum androgen levels and muscle mass in women with polycystic ovary syndrome.

OBJECTIVE: To relate serum androgen levels and muscle size in women with polycystic ovary syndrome (PCOS). METHODS: Forty-eight women with PCOS were studied. Baseline characteristics included age, height, weight, and hirsutism. Total-body lean mass was measured by wholebody scanning with dual-energy x-ray absorptiometry, and the lean mass index (total-body lean mass/height2, kg/m2) was calculated. Trunk-leg fat mass ratio (trunk-leg fat ratio) was assessed by dual-energy x-ray absorptiometry. Serum testosterone, dehydroepiandrosterone sulfate, and androstenedione levels were measured by radioimmunoassay. Subjects were arbitrarily divided into two groups according to lean mass index (lean mass index at least 14 kg/m2, n = 24; or less than 14 kg/m2, n = 24). Baseline characteristics and serum androgen levels were compared between the groups. In all subjects, lean mass index was correlated with age, height, trunk-leg fat ratio, and serum androgen levels. RESULTS: Although trunk-leg fat ratio, serum testosterone and androstenedione levels, and the prevalence of hirsutism were significantly higher in women with a lean mass index at least 14 kg/m2, age and height did not differ significantly between the groups. Trunk-leg fat ratio and serum testosterone and androstenedione levels correlated significantly with lean mass index (r = .68, .75, and .54, respectively). Hirsutism also correlated with lean mass index (standardized regression coefficient = .49; P<.05). CONCLUSION: Muscle size in women with PCOS correlated positively with serum androgen levels and upper body-fat distribution, independent of height.

Adult↗
PubMed1999

Apoptosis in placentas from human T-lymphotropic virus type I-seropositive pregnant women: a possible defense mechanism against transmission from mother to fetus.

OBJECTIVE: The mechanism by which the placenta serves as the barrier against mother-to-fetus transmission of microorganisms remains to be elucidated. Programmed cell death, apoptosis, is considered a cellular defense mechanism against infection. The hypothesis of this study is that apoptosis of human T-lymphotropic virus type I (HTLV-I)-infected placental villous cells is involved in the defense mechanism against mother-to-fetus transmission of HTLV-I. METHODS: Apoptosis was compared in term placentas from eight HTLV-I-seropositive pregnant women and eight HTLV-I-seronegative pregnant women by the terminal deoxynucleotidyl transferase-mediated deoxyuridine nick end-labeling method. In addition, an in vitro cocultivation with an HTLV-I-infected lymphocyte cell line (MT-2 cells) was performed to examine whether placental villous cells were infected with HTLV-I and apoptosis was induced. RESULTS: The incidence of apoptosis-positive cells (nuclei) in placentas from the HTLV-I-seropositive pregnant women was higher than in the HTLV-I-seronegative pregnant women (P < .02). Cocultivation with MT-2 cells showed that trophoblast cells were able to be infected with HTLV-I and that apoptosis was induced in the placental villous cells. CONCLUSION: HTLV-I infection induces apoptosis in the placenta. We speculate that apoptosis may be involved in the defense mechanism of the placenta against mother-to-fetus transmission of HTLV-I.

Adult↗
PubMed1999

Differences in target outline delineation from CT scans of brain tumours using different methods and different observers.

PURPOSE: To assess errors resulting from manual transfer of contour information for three-dimensional (3-D) target reconstruction, and to determine variations in target volume delineation of brain tumours by different radiation oncologists. MATERIALS AND METHODS: Images of 18 patients with intracranial astrocytomas were used for retrospective treatment planning by five radiation oncologists. In this study, the target outline was delineated on sequential CT slices by an experienced radiation oncologist. Thereafter, the target outline was manually reconstructed by five radiation oncologists onto an A-P or lateral scout film. The same target outline was also reconstructed as a projection using the Beam's-eye view capability on a CT simulator unit. The two target outlines were compared by encompassing each shape with the smallest rectangle. The manually-reconstructed radiation field was termed 'Field manually established on X-ray film (F-X)', and the automatically-established field was termed 'Field established by CT simulator (F-CT)'. In a second part of this study, four radiation oncologists defined contours from contrast enhanced CT images of nine patients with intracranial astrocytomas. The CT images of these nine cases included five pre-operative cases and four post-operative cases. Both gross tumour volume (GTV) and clinical target volume (CTV) were outlined on sequential CT slices. The target outlines for the four radiation oncologists were compared by identifying the smallest rectangular field surrounding the projection of these contours. The field established by each radiation oncologist was termed 'Field of target volume (F-TV)', and the overlapping portion of the four F-TVs for each case was termed 'Overlapped field of the target volume (Fo-TV)'. RESULTS: The average distance between the isocentres of F-X and F-CT was 0.6 +/- 0.4 cm (mean +/- SD). The average ratio of the area of F-X divided by the area of F-CT was 1.04 +/- 0.12. The area of F-X was wider than the area of F-CT for four of the five oncologists. The ratio of the area of F-TV divided by the area of Fo-TV was calculated. The average ratio was relatively greater for CTV (2.07 in pre-operative cases and 2.11 in post-operative cases) than for GTV (1.12 in pre-operative cases and 1.41 in post-operative cases). Among radiation oncologists, variations in the delineation of GTV were smaller than those of CTV. CONCLUSIONS: When using an X-ray simulator in treatment planning, errors resulting from the manual transfer of CT contour information to planar radiographs must be considered. When computer techniques are used to project contours onto radiographs errors resulting from individual variations when performing the contouring must be considered.

Astrocytoma↗
PubMed1999

A novel panoramic machine for photofluorography.

OBJECTIVE: To describe the main features of a prototype panoramic X-ray machine modified for photofluorography. METHODS: The prototype PTM 2001 (Asahi Roentgen, Kyoto, Japan) consists of a conventional panoramic machine with a modified X-ray generator and incorporating a 9-inch image intensifier and a photofluorographic camera. RESULTS: The photofluorographic panoramic radiograph is reduced in size by 60% compared with conventional images. Using 10 cm x 45.7 m photoradiographic film 250 panoramic images can be continuously obtained. Examination time is shorter since cassette changes are not necessary. The radiation dose-in-air, measured in front of the secondary collimator, is 174 muGy. CONCLUSIONS: The advantages of the photofluorographic panoramic radiography include cost economy, shorter examination time and lower radiation exposure.

Equipment Design↗
PubMed1999

Complete analysis of genes and enzymes for gamma-hexachlorocyclohexane degradation in Sphingomonas paucimobilis UT26.

gamma-Hexachlorocyclohexane (gamma-HCH; also called BHC or lindane) is one of the highly chlorinated pesticides which can cause serious environmental problems. Sphingomonas paucimobilis UT26 degrades gamma-HCH under aerobic conditions. The unique degradation pathway of gamma-HCH in UT26 is revealed. In the upstream pathway, gamma-HCH is transformed to 2,5-dichlorohydroquinone (2,5-DCHQ) by two different dehalogenases (LinA and LinB) and one dehydrogenase (LinC) which are expressed constitutively. In the downstream pathway, 2,5-DCHQ is reductively dehalogenated, and then ring-cleaved by enzymes (LinD and LinE, respectively) whose expressions are regulated. We have cloned and sequenced five structural genes (linA, linB, linC, linD, and linE) directly involved in this degradation pathway. The linD and linE genes form an operon, and its expression is positively regulated by the LysR-type transcriptional regulator (LinR). The genes linA, linB, and linC are constitutively expressed, and are present separately from each other in the UT26 genome. Cell fractionation analysis, Western blotting, and immuno electron microscopy revealed that LinA and LinB are localized in the periplasmic space of UT26.

Journal Article↗
PubMed1999

Evaluation of endoscopic ultrasonography as an indicator for surgical treatment of gastric cancer.

BACKGROUND AND METHODS: Clinicopathological analysis of 346 patients with gastric cancer was made retrospectively and new criteria for the indication of a limited operation using endoscopic ultrasonography (EUS) was developed. Suggested new criteria for selecting gastric cancer patients for the limited operation were: (i) the cancer is located in the mucosa and the lymph nodes are not involved as indicated by EUS examination; (ii) the maximum size of the tumour is less than 2.0 cm; (iii) there are no multiple gastric cancers or simultaneous abdominal cancers; and (iv) the mucosal cancer of elevated type less than 2.0 cm is excluded because there are good indications for endoscopic mucosal resection. RESULTS AND CONCLUSIONS: We applied these new criteria to 262 patients and found that the patients who had limited operation had the same prognosis and a better quality of life compared with patients who had the conventional operation.

Adult↗
PubMed1999

Identification and characterization of a new intermediate in the ribosylative inactivation pathway of rifampin by Mycobacterium smegmatis.

Mycobacterium smegmatis DSM 43756 inactivates rifampin by ribosylation. To study this process of rifampicin, all possible inactivated forms of the antibiotic were extracted and purified. Structural studies showed the presence of a new inactivation product, designated RIP-TAp(23-phosphoribosyl-rifampin). Formation of 23-(O-ADP-ribosyl)rifampin (RIP-TAs) is the first step, followed by removal of AMP to give rise to the newly identified compound. Lastly, dephosphorylation leads to formation of 23-ribosyl-rifampin (RIP-Mb). Feeding experiments with the ADP-ribosylated antibiotic obtained from the cell homogenates of an Escherichia coli strain carrying the cloned M. smegmatis gene confirmed this rifampin inactivation process.

Antibiotics, Antitubercular↗
PubMed1999

Clinical results of transcatheter arterial infusion for uterine cervical cancer.

The effects of transcatheter intraarterial infusion of anticancer drugs on the prognosis of cervical cancer were retrospectively studied. Two or three sessions of transcatheter arterial infusion therapy were performed for 68 patients with primary uterine cervical cancer. The number of patients with stage I, II, III, or IV disease were 13, 22, 24, and 9, respectively. Patients with squamous cell carcinoma comprised 3, 17, 17, and 5 of the respective groups, and the patients with stage I and II disease had either adenocarcinoma or adenosquamous carcinoma, or bulky tumor (>4 cm). The drugs infused were cisplatin (60-70 mg/m2), doxorubicin hydrochloride (30-40 mg/m2), mitomycin (15 mg/m2), and 5-fluorouracil (500 mg/body). They were infused via the bilateral internal iliac arteries. Fifty-eight of the 68 patients (85%) received a radical hysterectomy after transcatheter arterial infusion: 12 of 13 with stage I disease, 21 of 22 with stage II disease, 20 of 24 with stage III disease, and five of nine with stage IV disease. Two patients with stage III disease received radical radiotherapy. The other eight patients (one with stage I disease, one with stage II disease, two with stage III disease, and four with stage IV disease) did not receive an operation after transcatheter arterial infusion because they had distant metastases at the time of operation. Thirty-two of 58 patients (56%) received postoperative radiotherapy. The complete histologic response rates (no active cancer cells) after transcatheter arterial infusion were: 2 of 12 patients with stage I disease, 3 of 21 patients with stage II disease, 5 of 20 patients with stage III disease, and one of five patients with stage IV disease. Tumors with squamous cell carcinoma disappeared at a significantly better rate (10/36, 28%) than did tumors with adenocarcinoma or adenosquamous cell carcinoma (1/22, 5%; p < 0.05). The overall 5-year survival rates of the patients with stages I, II, and III disease were 92.3%, 62.2%, and 71%, respectively. The 5-year survival rates of the patients who underwent surgery with stage I, II, and III disease were 100%, 66.3%, and 71.5%, respectively. Leukocytopenia and thrombocytopenia occurred as an acute complication in 75% and 79% of the patients, respectively. As a late complication, ileus occurred in 7%. Transcatheter arterial infusion may improve the prognosis of patients with cervical cancer without increasing the incidence of late complications.

Antineoplastic Combined Chemotherapy Protocols↗
PubMed1999

Crystallization and preliminary X-ray diffraction analysis of haloalkane dehalogenase LinB from Sphingomonas paucimobilis UT26.

Haloalkane hydrolytic dehalogenase LinB from Sphingomonas paucimobilis UT26, an enzyme which releases chloride or bromide anion from n-halogenated alkanes and has a broad range of substrate specificity, was crystallized using the hanging-drop vapour-diffusion method at 278 K. The best crystals were obtained by microseeding with a precipitant containing 18-20%(w/v) PEG 6000, 0.2 M calcium acetate and 0.1 M Tris-HCl pH 8.9. The crystals diffract to at least 1.60 A using synchrotron X-ray under cryogenic (100 K) conditions. They belong to the orthorhombic space group P21212 with unit-cell parameters a = 50.29, b = 71.70, c = 72.73 A. The asymmetric unit contains one molecule of the enzyme.

Crystallization↗