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Biomedical subjects

S Inoue

Publications and source records attributed to S Inoue.

At least 631 records · Page 35Linked to original sources
PubMed1996

Substrate specificity of rainbow trout testis CMP-3-deoxy-D-glycero-D-galacto-nonulosonic acid (CMP-Kdn) synthetase: kinetic studies of the reaction of natural and synthetic analogues of nonulosonic acid catalyzed by CMP-Kdn synthetase.

In this report we present kinetic data of the activation reaction of several synthetic 3-deoxy-D-glycero-D-galacto-nonulosonic acid (Kdn) and N-acetylneuraminic acid (Neu5Ac) analogues catalyzed by the rainbow trout testis CMP-Kdn synthetase. This enzyme showed broad substrate specificity in terms of substitutions at C4 or C5 position of Kdn and Neu5Ac. In contrast, calf brain CMP-N-acylneuraminic acid synthetase had narrow substrate specificity, being active only on various N-acyl analogues of Neu5Ac and only slightly active on Kdn derivatives. Usefulness of the trout testis enzyme for synthesis of various CMP-sialate analogues, which could be donor substrates for sialyltransferases, was demonstrated.

Animals↗
PubMed1996

Identification of a novel isoform of estrogen receptor, a potential inhibitor of estrogen action, in vascular smooth muscle cells.

Clinical and experimental studies showed that estrogen has antiatherogenic effects. We previously demonstrated that the estrogen receptor (ER) mRNA and protein are expressed in vascular smooth muscle cells (VSMC) derived from rat aorta. Here, the expression of isoforms of the ER was examined in VSMC. Reverse transcriptase-polymerase chain reaction using specific primers for rat ER cDNA was performed from RNA of rat VSMC. This revealed the existence of ER cDNA that is shorter than the wild-type ER cDNA. Sequencing of the amplified products identified three isoforms of the ER and the wild-type ER. These ER mRNA isoforms lacked the region corresponding to exon 4, exon 4 and 5, and exon 3 and 4. Therefore, they were designated as ERdelta4 isoform, ERdelta4/5 isoform and ERdelta3/4 isoform, respectively. Chloramphenicol acetyltransferase assay was performed with these ER isoforms constructed into the expression vector and the reporter plasmid containing the estrogen responsive element. The assay showed that these ER isoforms lost estrogen-dependent transactivation activities and that ERdelta4/5 isoform has a inhibitory effect on normal estrogen action when it was cotransfected with the wild-type ER. These ER isoforms might be involved in the regulation of VSMC by estrogen.

Amino Acid Sequence↗
PubMed1996

Induction, localization, and purification of a novel sialidase, deaminoneuraminidase (KDNase), from Sphingobacterium multivorum.

Recently, we reported the discovery of a new type of sialidase, KDNase, which specifically hydrolyzes the ketosidic linkages of 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid (KDN), but not N-acylneuraminyl linkages. We now report that this enzyme, designated KDNase SM, is an inducible enzyme that is localized in the periplasm of Sphingobacterium multivorum. Growth of S. multivorum in the presence of KDN-containing oligosaccharide alditols, KDNalpha2-->3Galbeta1-->3GalNAc alpha1-->3[KDNalpha2--> (8KDN alpha2-->)n-->6]GalNAcol, as a sole carbon source induced KDNase SM activity 15 40-fold, compared with growth in the absence of inducer. KDN, Neu5Ac, or Neu5Ac oligomers were ineffective as inducers. The enzyme was released from the periplasm of induced cells by cold osmotic shock and purified 700-fold to homogeneity. The specific activity of the pure enzyme was 82,100 units/mg of protein. KDNase SM activity resided in a single polypeptide chain with an estimated molecular weight of approximately 47,500. Enzyme activity was maximal at near neutral pH. The availability of pure KDNase will now make it possible to study the structure and functional role of KDN-glycoconjugates and to determine the molecular mechanism whereby the enzyme can discriminate between KDN and N-acylneuraminic acid.

Chromatography, Ion Exchange↗
PubMed1996

Construction of highly extensive polymorphic DNA libraries by in-gel competitive reassociation procedure.

Differential genomic DNA libraries between two mouse strains and from two human individuals were constructed by means of the in-gel competitive reassociation (IGCR) procedure, a procedure developed for cloning altered anonymous restriction fragments. The libraries were highly enriched in RFLP fragments, approximately 60 and 40% for the mouse and human libraries, respectively, and, more importantly, maintained most of the original complexities of the RFLP fragments. Therefore, differential genomic DNA libraries constructed by the IGCR procedure, particularly for human genomic DNA, should offer highly extensive sources for polymorphic DNA sequences necessary for a variety of genome analyses, including studies on the origin and mechanism of biological diversity among the same species.

Animals↗
PubMed1996

Ultrastructural characterization of internal basement membrane of junctional epithelium at dentogingival border.

BACKGROUND: Although a number of electron microscopic observations of the dentogingival border have been reported, observation of the detailed ultrastructure of the internal basement membrane of the junctional epithelium at the border has not yet been done. METHODS: The internal basement membrane of a rhesus monkey (Macaca mulatta) was ultrastructurally characterized with high-resolution electron microscopy. The head and neck region of a 5-year-old animal was perfused with a fixative containing 1.25% glutaraldehyde, 0.75% formaldehyde, and 0.04% CaCl2 in 0.1 M phosphate buffer, pH 7.4. Isolated mandibular segments were further fixed with 2.5% glutaraldehyde, 2% formaldehyde, and CaCl2 (300 mg/l) in 0.1 M sodium cacodylate buffer, pH 7.4, for 24 hr at 4 degrees C. The segments were then demineralized and cut into small pieces that were postfixed with 1% osmium tetroxide and further processed for electron microscopy. RESULTS: The internal basement membrane takes the form of both a common basement membrane and, in places, a multilayered basement membrane. This basement membrane was found to have fundamental ultrastructural features common to other basement membranes. That is, the bulk of the basement membrane is composed of a network of irregular anastomosing strands referred to as "cords." Heparan sulfate proteoglycan-containing ribbonlike 4.5-5-nm-wide "double tracks" were also present in this basement membrane. However, the lamina densa of this basement membrane was unusually thick (160 nm) compared with the thickness (30-60 nm) in other basement membranes, and the cords were also much thicker (8.3 +/- 1.9 nm vs. the usual 3-5 nm). CONCLUSIONS: From these results, it is concluded that this particular basement membrane is specialized for mechanical strength. This specialization is likely to be suitable for the purpose of the tight sealing of the periodontal tissues from the oral environment.

Animals↗
PubMed1996

Association of bone mineral density with polymorphism of the estrogen receptor gene.

PvuII and XbaI restriction fragment length polymorphisms (RFLPs) of the estrogen receptor (ER) gene and its relation to bone mineral density (BMD) were examined in 238 postmenopausal healthy women aged 45-91 years (66.3 +/- 0.6 years, mean +/- standard error of the mean [SEM]) in Japan. The RFLPs were represented as Pp (PvuII) and Xx (XbaI), with capital letters signifying the absence of and small letters the presence of restriction sites. In the PPxx genotype (n = 18), Z score values of BMD were significantly lower than those for other genotypes (n = 220) (lumbar spine, -0.746 vs. -0.065 [p = 0.022]; total body, -0.482 vs. 0.308 [p = 0.002]). We classified the subjects into three genotypes with allelic haplotype: homozygote of the Px haplotype was expressed as the 11 genotype, heterozygote of the Px haplotype as the 10 genotype, and the one lacking the Px haplotype as the 00 genotype. The PpXx genotype was not included in this analysis because the allelic haplotypes are uncertain. The Px haplotype was associated with a low BMD in postmenopausal women (Z score for the lumbar spine, -0.746 vs. -0.279 vs. 0.083, for the 11, 10, 00 genotypes, respectively [p = 0.029]; Z score for the total body, -0.482 vs. 0.164 vs. 0.427, respectively [p = 0.003]). We suggest that some variation of the ER gene linked to these RFLPs is associated with low BMD and that this at least partly explains the cause of postmenopausal osteoporosis in Japanese women.

Aged↗
PubMed1996

Nephritogenicity and alpha-chain composition of NC1 fractions of type IV collagen from bovine renal basement membrane.

Nephritogenicity (anti-GBM-nephritis-inducing activity) and alpha-chain composition of globular-do-main (NCI) fractions of type IV collagen from bovine renal, pulmonary, and placental basement membranes (BMs) was examined by injecting these fractions with adjuvant into WKY/NCrj rats and by Western blotting using epitope-defined monoclonal antibodies to the six different alpha chains of type IV collagen. A purified nephritogenic fraction from renal BM contained alpha 1-alpha 6(IV)NCI, whereas a non-nephritogenic fraction contained only alpha 1-alpha 2(IV)NCI. Renal and pulmonary NCI had strong nephritogenic activity: placental NCI had weak activity. The renal and pulmonary fractions contained alpha 1-alpha 6(IV)NCI, and the placental fraction had a large amount of alpha 1-alpha 2(IV)NCI and a very small amount of alpha 3-alpha 6(IV)NCI. Immunohistochemical study of bovine renal BM with the monoclonal antibodies revealed that bovine glomerular BM contained alpha 1-alpha 5(IV) chains, but not the alpha 6(IV) chain. The absence of alpha 6(IV) chain in glomerular BM in bovine and in humans indicates that alpha 6(IV) chain is not a target antigen of anti-GBM nephritis. Nephritogenicity is apparently a property of alpha 3-alpha 5(IV)NCI.

Animals↗
PubMed1996

Isolation and identification of novel sulfated and nonsulfated oligosialyl glycosphingolipids from sea urchin sperm.

Novel sulfated and nonsulfated oligosialylglycosphingolipids were isolated from sperm of the sea urchin, Hemicentrotus pulcherrimus, and their structures were established as follows: [formula: see text] This provides the first evidence for the natural occurrence of a tetrasialic acid structure in glycosphingolipids. The finding of sulfated oligosialyl chains is especially noteworthy in that the sulfate group exclusively resides on the C-8 of the nonreducing terminal residues of oligo/polysialyl chains and that sulfation appears to be a termination signal for elongation of oligosialyl chains. Sulfation at the nonreducing terminal Neu5Ac residues of oligosialyl chains was also found to facilitate the formation of an inter-residue lactone between the carboxyl group at the nonreducing terminal sulfated Neu5Ac and the hydroxyl group at C-9 of the penultimate Neu5Ac residue. The long chain base was 4-hydroxysphinganine (t18:0) and the major fatty acid species were identified as C20:1, C21:1, and C22:1.

Animals↗
PubMed1996

Extrapancreatic nerve plexus invasion by carcinoma of the head of the pancreas. Diagnosis with intraportal endovascular ultrasonography.

CONCLUSION: The intraportal endovascular ultrasonography (IPEUS) could diagnose the second portion of the extrapancreatic nerve plexus invasion and provide precise information in operative strategy. But, the first portion was not visualized clearly owing to poor tissue penetration of the ultrasound beam, which may have reduced diagnostic accuracy. Improvement of the scanning area is expected to make intraportal endovascular US even more useful. BACKGROUND: Pancreatic cancer easily invades the retroperitoneal tissue, especially the extrapancreatic nerve plexus. We evaluated the extrapancreatic nerve plexus invasion of the pancreatic cancer with IPEUS. IPEUS was performed intraoperatively in 20 consecutive resected cases with carcinoma of the head of the pancreas. METHODS: IPEUS was performed with an 8-French, 20 MHz intravascular ultrasound catheter. IPEUS visualized the inferior pancreaticoduodenal artery (IPDA) in the extrapancreatic nerve plexus. The high-echoic area around the IPDA corresponds to the second portion of the extrapancreatic nerve plexus. The sonographic criterion for detection of the extrapancreatic nerve plexus invasion is low-echoic infiltration around the IPDA. RESULTS: Extrapancreatic nerve plexus invasion was confirmed with resected specimens in 10 patients. The IPDA could not be visualized in two patients. In 18 patients, the diagnostic accuracy of invasion was evaluated. For diagnosis of extrapancreatic nerve plexus invasion with intraportal endovascular US, the sensitivity, specificity, and overall accuracy were 87.5, 90, and 88.7%, respectively.

Adult↗
PubMed1996

Failed spinal anaesthesia: cause identified by MRI.

PURPOSE: To determine the cause of failed spinal anaesthesia, magnetic resonance images of the lumbar spine were examined. CLINICAL FUTURES: A 28-yr-old woman, scheduled for closure of a rectal fistula under spinal anaesthesia, hyperbaric tetracaine 12.5 mg injected into the subarachnoid space at L3-4 repeatedly resulted in restricted sacral spread. Magnetic resonance imaging showed that the cylindrical dimension of the lumbar spinal canal was wider than in normal subjects, suggesting a larger volume of CSF within the dural sac below the termination of the cord. The summit of the lumbar convexity was located at L3, which was more cephalad than the L3-4 puncture site. CONCLUSION: The uncommon anatomical characteristics of the lumbar spinal canal identified by magnetic resonance imaging were considered to be the reason for the restricted sacral spread observed repeatedly in this patient.

Adult↗
PubMed1996

Sevoflurane anaesthesia for a patient with adult polyglucosan body disease.

PURPOSE: Adult polyglucosan body disease (APBD) is a rare neurological disorder of unknown cause characterized by four manifestations: upper motor neuron signs, peripheral neuropathy with motor and sensory loss, urinary incontinence, and dementia. The purpose of this report is to present a patient with APBD anaesthetized successfully with sevoflurane and nitrous oxide. CLINICAL FEATURE: A 51-yr-old man with APBD was scheduled for haemorrhoidectomy. Paraesthesia, dysaesthesia, distal muscular atrophy and fasciculation were recognized in the extremities. Dementia, bulbar paralysis and respiratory insufficiency were basent. Anaesthesia was induced with inhalation of sevoflurane and nitrous oxide, and the trachea was intubated without the use of muscle relaxants. Maintenance of anaesthesia was performed with sevoflurane (inspired concentration: 1.5-2.5%) and nitrous oxide (50%). Emergence from anaesthesia and the postoperative course were uneventful, and no exacerbation of neurological signs and symptoms was recognized. No postoperative analgesia was required. CONCLUSION: General anaesthesia and tracheal intubation with sevoflurane and nitrous oxide provided safe anaesthesia for a patient with APBD.

Anesthesia↗
PubMed1996

Immunolocalization of transforming growth factor-beta in the bone tissue.

Fetal mouse calvarias were cultured in a medium containing fetal calf serum, 1,25(OH)2D3 and parathyroid hormone in 5% CO2 incubator at 37 degrees C for 5 days. The calvarias were then fixed in a buffered paraformaldehyde solution and the periostea were removed. A polyclonal antibody against transforming growth factor-beta (TFG-beta) and a second antibody labeled with fluorescein isothiocyanate were used to detect TGF-beta. As a result, a specific staining was observed only in the shielding zone covered by osteoclasts in the bone resorption lacunae and in the extracellular matrix adjacent to the osteoclasts. These findings suggested the local delivery and possible activation of TGF-beta by osteoclasts in tissue.

Animals↗
PubMed1996

Inhibition by triptoquinone-A of LPS- and IL-1 beta-primed induction of NO synthase in rat thoracic aorta.

We investigated the effect of triptoquinone-A (TQA), an active principal of Triptergium wilfordii, on the induction of nitric oxide synthase (NOS) promoted by endotoxin (LPS) and interleukin-1 beta (IL-1 beta). Prophylactic application of TQA selectively prevented LPS-primed initiation of L-arginine (Arg)-induced relaxation, and cGMP formation of rat thoracic aorta, and LPS-stimulated nitrite production by cultured aortic smooth muscle cells, which appear to be mediated by NOS expressed by LPS in vascular smooth muscle. TQA also prevented IL-1 beta-triggered initiation of Arg-induced relaxation and nitrite accumulation. These results suggest that TQA prevents LPS-or IL-1 beta-promoted induction of NOS in vascular smooth muscle, thus inhibiting development of Arg-induced vasorelaxation.

Animals↗
PubMed1996

Expression of CD26/dipeptidyl peptidase IV in adult T cell leukemia/lymphoma (ATLL).

The association of CD26/dipeptidyl peptidase IV (DPPIV) and human T lymphotropic virus type I (HTLV-I) was studied by two approaches. First, we examined the expression of CD26 in peripheral blood mononuclear cells (PBMC) from the patients with adult T cell leukemia/lymphoma (ATLL), an HTLV-I-related malignancy. The expression of CD26 on the surface of PBMC was decreased in all 20 patients with ATLL compared with those from normal individuals (P < 0.01) and the expression of the CD26 gene transcript was not detectable in seven out of eight patients with ATLL. Then we compared the quantity of viral DNA in CD26-negative (CD26-) and CD26-positive (CD26+) cells obtained from 17 HTLV-I healthy carries by using a polymerase chain reaction method. The CD26-cells had a higher copy number of viral DNA than CD26+ cells. These findings indicate that HTLV-I has in vivo tropism to CD26- cells, suggesting that some phenotypes of ATLL cells reflect the in vivo cellular tropism of HTLV-I.

Adult↗
PubMed1996

Reduced hydrogen peroxide production in neutrophils from patients with diabetes.

We evaluated the effects of granulocyte colony-stimulating factors (G-CSF), granulocyte-macrophage colony-stimulating factors (GM-CSF) and phorbol myristate acetate (PMA) on H2O2 production in purified neutrophils from patients with diabetes mellitus (DM), using flow cytometry. Twenty-two age-matched male subjects were selected: 11 were normal volunteers and the remainder had DM. No significant differences in intracellular H2O2 production per 60 min was observed in the "resting' neutrophils from DM patients (37.2 +/- 20.4 A.U.) compared with those from normal volunteers (24.9 +/- 8.4 A.U.). The PMA-stimulated neutrophils from normal volunteers generated approximately 4-fold increases in H2O2 per 60 min compared with those from DM patients. Under similar culture conditions, G-CSF caused 1.6-fold increases of H2O2 in neutrophils from normal volunteers compared with those of DM patients. Increases after GM-CSF stimulation were 2-fold higher in volunteer neutrophils compared with those from DM patients. The levels of G-CSF- or GM-CSF-stimulated H2O2 production in neutrophils from DM patients were low and were little different from non-stimulated resting cells. These data showed that H2O2 production in neutrophils induced by PMA is impaired in patients with DM, and neither G-CSF nor GM-CSF enhances its production.

Adult↗
PubMed1996

Sedation for symptom control in Japan: the importance of intermittent use and communication with family members.

We reviewed the circumstances surrounding the use of sedation for symptom control in a Japanese hospice. Of 143 inpatients, 69 (48.3%) received sedation and died an average 3.9 days after sedation was begun. Symptoms requiring sedation included dyspnea, pain, general malaise, agitation, and nausea. In 83% of cases, those symptoms were escalating as death approached. In about one-half of the cases, sedation was carried out intermittently until the patient died. Sedation was gained by such sedatives as midazolam, morphine, and haloperidol. Side effects included suppression of the respiratory and/or circulatory system in nine cases (in four cases it caused death), and delirium in one case; tolerance and dependence were also observed in two cases. We also examined the explanation to and understanding of the patients and their family members about sedation. This experience suggested the type of communication methods that are likely to be useful in Japan. It stresses the importance of intermittent use of sedation and communication with family members. We propose criteria for sedation to improve symptom control that would be acceptable in Japan.

Aged↗
PubMed1996

L-arabinose selectively inhibits intestinal sucrase in an uncompetitive manner and suppresses glycemic response after sucrose ingestion in animals.

The objective of this study was to investigate the effects of L-arabinose on intestinal alpha-glucosidase activities in vitro and to evaluate its effects on postprandial glycemic responses in vivo. L-Arabinose inhibited the sucrase activity of intestinal mucosa in an uncompetitive manner (Ki, 2 mmol/L). Neither the optical isomer D-arabinose nor the disaccharide L-arabinobiose inhibited sucrase activity, whereas D-xylose was as potent as L-arabinose in inhibiting this activity. L-Arabinose and D-xylose showed no inhibitory effect on the activities of intestinal maltase, isomaltase, trehalase, lactase, and glucoamylase, or pancreatic amylase. In contrast, a known alpha-glucosidase inhibitor, acarbose, competitively inhibited (Ki, 1.1 mumol/L) sucrase activity and also inhibited intestinal maltase, glucoamylase, and pancreatic amylase. L-Arabinose suppressed the increase of blood glucose after sucrose loading dose-dependently in mice (ED50, 35 mg/kg), but showed no effect after starch loading. The suppressive effect of D-xylose on the increase of blood glucose after sucrose loading was 2.4 times less than that of L-arabinose, probably due to intestinal absorption of the former. Acarbose strongly suppressed glycemic responses in both sucrose loading (ED50, 1.1 mg/kg) and starch loading (ED50, 1.7 mg/kg) in mice. L-Arabinose suppressed the increase of plasma glucose and insulin in rats after sucrose loading, the suppression of the former being uninterruptedly observed in mice for 3 weeks. Thus, the results demonstrated that L-arabinose selectively inhibits intestinal sucrase activity in an uncompetitive manner and suppresses the glycemic response after sucrose ingestion by inhibition of sucrase activity.

Acarbose↗