Biomedical subjects

M Wada

Publications and source records attributed to M Wada.

At least 253 records · Page 14Linked to original sources

PubMed1998

HPLC with fluorescence detection of urinary phenol, cresols and xylenols using 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride as a fluorescence labeling reagent.

A simple and sensitive HPLC method for the determination of phenolic compounds, i.e., phenol (Phe), cresols (Cres) and xylenols (Xyls), was developed. After a pre-column fluorescence derivatization of these compounds with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl) at 60 degrees C for 30 min, 11 DIB derivatives were successfully separated within 50 min with an ODS column using CH3CN-H2O-CH3OH (25 + 22 + 53, v/v) as the eluent. The detection limits of DIB derivatives at a signal-to-noise ratio of 3 ranged from 0.15 to 1.09 microM (0.2-1.6 pmol per 20 microliters). The precision of the proposed method for both within- and between-day assays of free and total phenol related compounds was satisfactory (RSD < 9.5%). By the proposed method, Phe and p-Cre could be detected in normal urine samples, and the calculated concentrations of free Phe and p-Cre in unhydrolysed urine samples were 1.5 +/- 1.3 and 23.9 +/- 24.3 microM and those of total Phe and p-Cre in hydrolysed urine samples were 87.3 +/- 81.2 and 200.7 +/- 195.4 microM (n = 21), respectively.

Benzoates↗

PubMed1998

NPS R-568 halts or reverses osteitis fibrosa in uremic rats.

Osteitis fibrosa is a common bone injury associated with secondary hyperparathyroidism (2(o)HPT). NPS R-568 is a phenylalkylamine derivative that acts as an agonist at the cell-surface Ca2+ receptor ("calcimimetic") and inhibits parathyroid hormone (PTH) secretion. In the present study, we tested whether NPS R-568 could ameliorate osteitis fibrosa in partially nephrectomized (Nx) rats with 2(o)HPT. Six months after surgery, Nx rats had developed mild but progressive 2(o)HPT and osteitis fibrosa. Two groups of Nx rats received NPS R-568 (3 and 30 mg/kg body wt x day) by daily gavage for 30 days, which led to a dose-related decrease in serum PTH levels and to a marked reduction in peritrabecular fibrosis (0.96 +/- 0.49% to < 0.1%). Furthermore, 2(o)HPT was associated with decreases in volumetric cortical bone mineral density (vCtBMD) and in cortical bone stiffness at the femoral midshaft. NPS R-568 significantly restored the deficits in vCtBMD and stiffness. These results indicate that NPS R-568 has beneficial effects on bones with osteitis fibrosa by normalizing serum PTH levels.

Aniline Compounds↗

PubMed1998

Effects of dietary n-3-to-n-6 polyunsaturated fatty acid ratio on mammary carcinogenesis in rats.

We investigated the effects of the dietary n-3-to-n-6 polyunsaturated fatty acid (PUFA) ratio (n-3/n-6 ratio) on mammary carcinogenesis induced by 7,12-dimethylbenz[a]anthracene in rats by feeding them several types of dietary fat with a fixed PUFA-to-saturated fatty acid ratio. Dietary fat was fed to the rats as 10% of the total feed weight, starting two weeks before the initiation. An increase in the n-3/n-6 ratio did not suppress the incidence or reduce the latency of mammary tumor development. The number and weight of mammary tumors per tumor-bearing rat tended to be large in the group with an n-3/n-6 ratio of 7.84 compared with those in the other groups. As the n-3/n-6 ratios were elevated, the total number and weight of tumors increased gradually. The prostaglandin E2 (PGE2) concentration in mammary tumor tissue was markedly low in the group with an n-3/n-6 ratio of 1.03 compared with the group with an n-3/n-6 ratio of 0.01. In addition, PGE2 concentrations were almost constant when n-3/n-6 ratios were > 1.03. These results suggested that the increase in the n-3/n-6 ratio of dietary fat with the fixed PUFA-to-saturated fatty acid ratio cannot suppress the mammary carcinogenesis but can promote development of tumors, despite reduced PGE2 concentration in the tumor.

9,10-Dimethyl-1,2-benzanthracene↗

PubMed1998

Mutations in the canilicular multispecific organic anion transporter (cMOAT) gene, a novel ABC transporter, in patients with hyperbilirubinemia II/Dubin-Johnson syndrome.

Members of the ATP-binding cassette (ABC) transporter superfamily are mutated to cause diseases that include cystic fibrosis, hyperinsulinemia, adrenoleukodystrophy, Stargardt disease and multidrug resistance. We recently isolated a novel human member of ABC transporter superfamily as the candidate transporter for the glucuronide and glutathione-conjugated antitumor agents, and found it highly homologous to the rat cmoat gene. consistent with recent findings of defects in the homologous cmoat gene in two rat models of hyperbilirubinemia (TR- and Eisai), we report two deletions and a missense mutation in the active transport family signature region in the gene in patients with hyperbilirubinemia II/Dubin-Johnson syndrome (DJS; MIM 237500), respectively. These results strongly implicate the cMOAT gene as responsible for the defects in DJS patients.

Amino Acid Sequence↗

PubMed1998

Relationship between gait and clinical results after high tibial osteotomy.

Thirty-two patients with primary osteoarthritis of the medial compartment of the knee were studied prospectively to assess the relationship between clinical results, limb alignment, and adduction moment of the knee. Clinical and radiographic examination and gait analyses were performed preoperatively and repeated at 6 months and at 1, 3, and 6 years after high tibial osteotomy. The preoperative peak adduction moment was high in 25 patients and low in seven. In both groups, the adduction moment of the knee decreased at 6 months after surgery but increased after that period. Alignment of the affected knee in both groups remained valgus after surgery (average femorotibial angle, 167 degrees-169 degrees). Clinical outcome in both groups improved after surgery and remained unchanged after 1 year. The peak adduction moment of the knee for the whole group significantly correlated with alignment and foot angle before and 6 years after surgery but did not correlate with stride length and walking velocity. In addition, only alignment was associated significantly with clinical results at 6 years. These results suggest that the preoperative peak adduction moment of the knee does not correlate with clinical or radiographic outcomes of high tibial osteotomy, provided sufficient valgus alignment is achieved at surgery.

Aged↗

PubMed1998

Sequences of ten circular ssDNA components associated with the milk vetch dwarf virus genome.

Milk vetch dwarf virus (MDV) is a member of the proposed genus Nanovirus, and its genome is composed of multiple, circular ssDNA components of about 1 kb. We have cloned and sequenced ten ssDNA components and designated them MDV-C1 to C10. Each DNA component contains one potential major open reading frame, and contains a putative stem-loop structure in the non-coding region. Notably, four components (C1, C2, C3 and C10) encode distinct replication-associated (Rep) proteins of 33 kDa, which show only limited (42-57%) amino acid identity. The six other components encode proteins with calculated molecular masses ranging from 12.7 to 19.7 kDa. Comparison of the sequences with those of other nanoviruses reveals that MDV is closely related to faba bean necrotic yellows virus (FBNYV) and subterranean clover stunt virus (SCSV). Six putative MDV genome products, including one Rep and five non-Rep proteins, show high (70.4-90.9%) amino acid identity to the corresponding six FBNYV proteins, whereas two other Rep proteins encoded by MDV-C2 and C3 are 82.3% and 73.0% identical to those encoded by SCSV-C2 and C6, respectively. These results indicate that MDV, FBNYV and SCSV have diverged from a common origin, which had multiple Rep components. In addition, the putative proteins encoded by MDV-C4 and its homologues contain a consensus retinoblastoma-binding motif, suggesting that they may be involved in controlling the host cell cycle.

Amino Acid Sequence↗

PubMed1998

Postoperative adjuvant radiotherapy and 5-fluorouracil chemotherapy for rectal carcinoma.

Postoperative combined modality therapy with radiotherapy and 5-fluorouracil (5FU) chemotherapy is an effective adjuvant approach that reduces locoregional and distant metastatic disease in patients with high-risk rectal carcinoma. However, this approach results in a treatment regimen of at least 6 months' duration. The present prospective study investigates the integration of radiotherapy and 5FU chemotherapy in a protocol designed to minimize toxicity and reduce the overall treatment time. A total of 40 patients with TNM stage II or III disease received postoperative radiotherapy at four fractions per week with weekly 5FU bolus injections delivered on the fifth non-radiotherapy day. Patients also received systemic chemotherapy with leucovorin both before and after pelvic irradiation, with the total treatment duration extending for only 18 weeks. Patients were able to complete radiotherapy in 90% of cases, while the delivery of full-dose chemotherapy was achievable in the vast majority. The incidence of haematologic and gastrointestinal toxicities requiring the cessation of treatment was acceptable. With a median follow-up of 20.9 months among surviving patients, the estimated progression-free and overall survival at 2 years were 71% and 79%, respectively.

Adenocarcinoma↗

PubMed1998

The P2 protein of rice dwarf phytoreovirus is required for adsorption of the virus to cells of the insect vector.

Intact particles of rice dwarf phytoreovirus adsorbed to and entered monolayer-cultured cells of the insect vector Nephotettix cincticeps and multiplied within the cells. Particles that lacked the P2 protein neither attached to nor infected such cells. Furthermore, P2-free particles obtained from a transmission-competent isolate of the virus were unable to infect insect vectors that had been allowed to feed on these virus particles through a membrane. However, when such virus particles were injected into insects via a glass capillary tube they successfully infected the insects, which became able to transmit the virus. These results support the hypothesis that, while P2-free particles can neither interact with nor infect cells in the intestinal tract of the insect vector, they do retain the ability to infect such cells when physically introduced into the hemolymph by injection.

Adsorption↗

PubMed1998

The 20-kilodalton (kDa) human growth hormone (hGH) differs from the 22-kDa hGH in the complex formation with cell surface hGH receptor and hGH-binding protein circulating in human plasma.

In spite of recent advance in understanding of the stoichiometry of 22-kDa human GH (22K-hGH) with cell surface hGH receptor (hGHR) and hGH-binding protein (hGH-BP) circulating in human plasma, that of 20-kDa hGH (20K-hGH) is poorly understood. To clarify this, mouse pro-B Ba/F3 cells stably expressing the full-length hGHR (Ba/F3-hGHR) and both recombinant and native hGH-BP were used in this study. Cell proliferation assay revealed that the two hGH isoforms increased Ba/F3-hGHR cells to the same extent in a dose-dependent manner at 0.1 pM-10 nM. However, the self-inhibition observed in 20K-hGH at 5 microM was significantly less than that in 22K-hGH. Furthermore, addition of 1 and 10 nM recombinant hGH-BP caused a slight inhibition in 20K-hGH, but a drastic inhibition in 22K-hGH. Gel filtration chromatography of mixtures of 20K-hGH with recombinant hGH-BP clearly demonstrated that 20K-hGH formed a 1:2 (hGH:hGH-BP) complex efficiently but no detectable 1:1 complex in any conditions. Supporting data were also obtained with native hGH-BP. Computer-aided homology modeling of 20K-hGH has provided speculative data that the conformational change caused by deletion of 15 residues may occur only in the loop between helix 1 and helix 2, resulting in the reduction of its site 1 affinity. In conclusion, 20K-hGH possesses a unique property for forming a 1:2 complex to the same extent as 22K-hGH but has difficulty in forming a 1:1 complex, which might be attributed to the conformational change restricted to its site 1 region.

Animals↗

PubMed1998

Escherichia coli transformant expressing the glucose dehydrogenase gene from Bacillus megaterium as a cofactor regenerator in a chiral alcohol production system.

Escherichia coli JM109 (pGDA2) overexpressing the glucose dehydrogenase (GDH) gene from Bacillus megaterium IWG3 was examined for use as a cofactor regenerator. In the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate by E. coli JM109 (pKAR) which is an aldehyde reductase-overproducing transformant, E. coli JM109 (pGDA2) can act as an NADPH regenerator with NADP+ and glucose, similarly to commercially available GDH.

Alcohols↗

PubMed1998

Purification and characterization of NADPH-dependent carbonyl reductase, involved in stereoselective reduction of ethyl 4-chloro-3-oxobutanoate, from Candida magnoliae.

A NADPH-dependent carbonyl reductase was purified to homogeneity from Candida magnoliae AKU4643 through four steps, including Blue Sepharose affinity chromatography. The enzyme catalyzed the stereoselective reduction of ethyl 4-chloro-3-oxobutanoate to the corresponding (S)-alcohol with a 100% enantiomeric excess, which is a useful chiral building block for the chemical synthesis of pharmaceuticals. The relative molecular mass of the enzyme was estimated to be 76,000 on high performance gel filtration chromatography and 32,000 on SDS polyacrylamide gel electrophoresis. The enzyme reduced alpha-, beta-keto esters and conjugated diketones in addition to ethyl 4-chloro-3-oxobutanoate. The enzyme activity was inhibited by quercetin and HgCl2, but not by EDTA. The N-terminal amino acid sequence of the enzyme showed no apparent similarity with those of other oxidoreductases.

Acetoacetates↗

PubMed1998

Growth hormone-induced tyrosine phosphorylation of EGF receptor as an essential element leading to MAP kinase activation and gene expression.

GH binding to its receptor, which belongs to the cytokine receptor superfamily, activates Janus kinase (JAK) 2 tyrosine kinase, thereby activating a number of intracellular key proteins such as STAT (signal transducers and activators of transcription) proteins and mitogen-activated protein (MAP) kinases, which finally lead to GH's biological actions including gene expression. In contrast to receptor tyrosine kinases, the signalling pathways leading to MAP kinase activation by GH are poorly understood but appear to involve Grb2 and Shc. We now show that GH stimulated tyrosine phosphorylation of epidermal growth factor receptor (EGFR) and its association with Grb2, and concomitantly stimulated MAP kinase activity in liver, a major target tissue. Expression of EGFR and its mutants into CHO-GH receptor (GHR) cells revealed that GH-induced full activation of MAP kinase and c-fos expression required tyrosine phosphorylation sites of EGFR but not its intrinsic tyrosine kinase activity. Moreover, by also using dominant negative JAK2 and in vitro kinase assay, we demonstrated that tyrosine 1068 of EGFR was evidently one of the major phosphorylation and Grb2 binding sites stimulated by GH via JAK2. These data suggest that the role of EGFR in GH signalling is to be phosphorylated by JAK2, thereby providing docking sites for Grb2 and activating MAP kinases and gene expression. This novel cross talk pathway may provide the first example of the hormone and cytokine receptor superfamily transducing signals via associated nonreceptor tyrosine kinase by phosphorylating growth factor receptor and utilizing it as a docking protein independent of its receptor tyrosine kinase activity.

Adaptor Proteins, Signal Transducing↗

PubMed1998

Changes in circulating LH, sex steroid hormones, thyroid hormones and corticosterone in relation to breeding and molting in captive humboldt penguins (Spheniscus Humboldti) kept in an outdoor open display.

Penguins are highly adapted to marine life. Their hydrodynamic efficiency depends on feathers which wear with age and need to be replaced regularly. During molting, penguins can not enter the sea to forage and are forced to fast. Therefore the duration of molting is necessarily brief. To better understand molting in penguins, we collected plasma samples from 16 (8 pairs) Humboldt penguins kept in an open display pen at Tokyo Sea Life Park from May to September, 1994 and estimated circulating concentrations of LH, testosterone, estradiol, thyroxine (T4), triiodothyronine (T3) and corticosterone. Body mass was also measured at each blood sampling. Throughout the year, reproductive activities (egg laying, incubation, hatching and rearing) and molting were observed and recorded. Humboldt penguins maintained reproductive activity from January to December except during molting. Each pair started molting between the end of July and early August; usually males started earlier. The duration of molting was 13.4 +/- 0.8 days for males and 12.9 +/- 0.3 days for females. Body masses were highest just before the start of molting in both sexes. Plasma concentrations of LH were high, (> 2 ng/ml) in May in both sexes, then gradually decreased, to 0.53 +/- 0.38 ng/ml in males and 0.72 +/- 0.11 ng/ml in females by the end of July. Testosterone and estradiol concentrations in plasma decreased and were lowest during molting. On the other hand, plasma concentrations of T4 were low until early July (ca. 20 ng/ml) and then doubled within 10 days; the high levels were maintained for one month and then decreased greatly in males and slightly in females. When the plasma concentrations of T4 started to decrease, plasma concentrations of LH increased. Changes in plasma T3 were not consistent with molting. These results indicate that the decrease of plasma levels of sex steroid hormones and the sharp increase of T4 induced molting, which lasted only for a short period.

Adaptation, Physiological↗

PubMed1998

New apparatus stimulating the region of tongue innervated by glossopharyngeal nerve and its application to monosodium glutamate (MSG) solution.

Some apparatuses stimulating the region of tongue innervated by chorda tympani nerve were well known and they were used popularly, but the apparatuses for glossopharyngeal nerve were not produced yet. In the study, the apparatus for the stimulations of glossopharyngeal nerve was trially produced. It was known that the substance of "Umami" was responded well in the tongue region innervated by glossopharyngeal nerve. The evoked potential for the stimulations of monosodium glutamate (MSG) solution which was known as substance of Umami was measured with the use of the trial stimulating apparatus. The evoked potential was detected by monopolar lead for the positions between Cz and A1 (left lobe) with the ground at Fpz. The evoked potential of MSG consisted of a positive peak wave with the latency at approximately 250 ms. The amplitudes increased and the latency decreased as the concentration of MSG increased. The evoked potential stimulating the tongue innervated by this glossopharyngeal nerve could be detected for the first time by the use of the trial apparatus.

Evoked Potentials↗

PubMed1998

Effect of indigestible saccharides on B lymphocyte response of intestinal mucosa and cecal fermentation in rats.

The effects of water-soluble and -insoluble indigestible saccharides (IDS) on immune responses of the intestinal tract were studied. Male 4-week-old Sprague Dawley rats were fed for three weeks on diets containing several kinds of IDS at 5%. The results revealed that the proportion of kappa-light chain and IgA-presenting lymphocytes in small intestinal and cecal mucosa differed in increased number depending on the type of IDS. The response of colonic mucosa was not pronounced. The amounts of short-chain fatty acid (SCFA) and lactic acid in the cecal contents of the other test groups except the celfur group tended to be higher than those in the cellulose group, particularly in the lactulose group where many acids showed significant increases. The correlation between the proportion of kappa-light chain and IgA-presenting lymphocytes in the cecal mucosa and lactic acid in the cecal contents was significant, but that between the proportion of both lymphocytes and SCFA was not. Based on the above, we concluded that the oral administration of IDS induces the proliferation of kappa-light chain and IgA-producing B lymphocytes in small intestinal and cecal mucosa, but the degree of response differs depending on the type of IDS. It is thus suggested that IDS are involved in the intestinal immune system of rats.

Animals↗

PubMed1998

Changes in platelet aggregation and lipid metabolism in rats given dietary lipids containing different n-3 polyunsaturated fatty acids.

We compared the effects of different n-3 polyunsaturated fatty acids (PUFA) on platelet aggregation and lipid metabolism in rats. alpha-Linolenic acid (ALA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were used as n-3 PUFA sources. The rats were fed diets containing 10% lipids (polyunsaturated/saturated fatty acid (P/S) ratio = 1.0; n-3/n-6 = 0.02 for the control group, 0.2 for the test groups) for two weeks. The platelet counts, platelet aggregation, and production of thromboxane A2 (TXA2), plasma total cholesterol (TC) and triacylglycerols (TG) were not different between the ALA group and the control group, but showed a decreasing tendency for the EPA group and significant decreases for the DHA group. The production of prostacyclin in the aorta was significantly decreased in all of the n-3 PUFA groups when compared with that in the control group. Liver TC and TG concentrations were significantly decreased in the DHA group when compared with those in the control group. Based on the above, it is assumed that the physiological action exerted by n-3 PUFA differs by type and that DHA is a more effective n-3 PUFA, both for suppressing platelet aggregation and for modulating lipid metabolism in the plasma and liver of rats.

Animals↗

PubMed1998

Functional evaluation of telemedicine with super high definition images and B-ISDN.

In order to determine whether a super high definition (SHD) image running at a series of 2048 resolution x 2048 line x 60 frame/sec was capable of telemedicine, we established a filing system for medical images and two experiments for transmission of high quality images were performed. All images of various types, produced from one case of ischemic heart disease were digitized and registered into the filing system. Images consisted of plain chest x-ray, electrocardiogram, ultrasound cardiogram, cardiac scintigram, coronary angiogram, left ventriculogram and so on. All images were animated and totaled a number of 243. We prepared a graphic user interface (GUI) for image retrieval based on the medical events and modalities. Twenty one cardiac specialists evaluated quality of the SHD images to be somewhat poor compared to the original pictures but sufficient for making diagnoses, and effective as a tool for teaching and case study purposes. The system capability of simultaneously displaying several animated images was especially deemed effective in grasping comprehension of diagnosis. Efficient input methods and creating capacity of filing all produced images are future issue. Using B-ISDN network, the SHD file was prefetched to the servers at Kyoto University Hospital and BBCC (Bradband ISDN Business chance & Culture Creation) laboratory as an telemedicine experiment. Simultaneous video conference system, the control of image retrieval and pointing function made the teleconference successful in terms of high quality of medical images, quick response time and interactive data exchange.

Computer Communication Networks↗

PubMed1998

Change of Chemosensory Event-Related Potentials on Olfactory Stimulation as a Function of Odorant Concentrations.

Change of chemosensory event-related potentials on olfactory stimulation as a function of odorant concentrations is discussed. According to the ascending method, from undetectable level to clearly detectable level, the dependency of shortening of the peak latency on the concentration of odorant was recognized as a rapid decrease of amplitude from the threshold level when results of our ascending trial were averaged. However, a much higher concentration of odorant did not always evoke a positive response and a shorter peak latency.

Journal Article↗