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Biomedical subjects

Shigeo Suzuki

Publications and source records attributed to Shigeo Suzuki.

35 records · Page 2Linked to original sources
PubMed2003

Existence of novel beta-1,2 linkage-containing side chain in the mannan of Candida lusitaniae, antigenically related to Candida albicans serotype A.

The antigenicity of Candida lusitaniae cells was found to be the same as that of Candida albicans serotype A cells, i.e. both cell wall mannans react with factors 1, 4, 5, and 6 sera of Candida Check. However, the structure of the mannan of C. lusitaniae was significantly different from that of C. albicans serotype A, and we found novel beta-1,2 linkages among the side-chain oligosaccharides, Manbeta1-->2Manbeta1--> 2Manalpha1-->2Manalpha1-->2Man (LM5), and Manbeta1-->2Man-beta1-->2Manbeta1-->2Manalpha1-->2Manalpha1-->2Man (LM6). The assignment of these oligosaccharides suggests that the mannoheptaose containing three beta-1,2 linkages obtained from the mannan of C. albicans in a preceding study consisted of isomers. The molar ratio of the side chains of C. lusitaniae mannan was determined from the complete assignment of its H-1 and H-2 signals and these signal dimensions. More than 80% of the oligomannosyl side chains contained beta-1,2-linked mannose units; no alpha-1,3 linkages or alpha-1,6-linked branching points were found in the side chains. An enzyme-linked immunosorbent inhibition assay using oligosaccharides indicated that LM5 behaves as factor 6, which is the serotype A-specific epitope of C. albicans. Unexpectedly, however, LM6 did not act as factor 6.

Antigens, Fungal↗
PubMed2003

Coxsackievirus and adenovirus receptor, a tight junction membrane protein, is expressed in glomerular podocytes in the kidney.

In nephrosis, filtration slits of podocytes are greatly narrowed, and slit diaphragms are displaced by junctions with close contact. Freeze-fracture studies have shown that the newly formed junctions consist of tight junctions and gap junctions. Several tight-junction proteins are known as integral membrane components, including occludin and claudins; but none of them have been found in podocytes. Coxsackievirus and adenovirus receptor (CAR) has recently been identified as a virus receptor that is a 46-kDa integral membrane protein with two Ig-like domains in the extracellular region. In polarized epithelial cells, CAR is expressed at the tight junction, where it associates with ZO-1 and plays a role in the barrier to the movement of macromolecules and ions. In the present study, we investigated the expression and localization of CAR in rat kidneys treated with puromycin aminonucleoside (PAN) and in rat kidneys perfused for 15 minutes with protamine sulfate (PS). Both the experimental models have been used to induce tight junctions in podocytes. Ribonuclease protection assay and Western blot analysis revealed a distinct increase of CAR transcript and protein in glomeruli during PAN nephrosis but no increase in glomeruli by PS perfusion. Immunohistochemistry revealed a significant increase in CAR staining intensity along the glomerular capillary wall in PAN nephrosis and after PS perfusion. Immunoelectron microscopy demonstrated in both the models that the immunogold particles for CAR along the capillary wall were found predominantly at close cell-cell contact sites of podocytes but were rarely found at slit diaphragms. In cultured podocytes, CAR was localized at cell-cell contact sites. CAR distribution was identical to that of ZO-1 and different from that of a gap junction protein, connexin43. These findings indicate that CAR is an integral membrane component of tight junction in podocytes and that CAR expression in podocytes is regulated at the transcriptional level and in the redistribution of protein.

Amino Acid Sequence↗
PubMed2003

Comparative study of protective effects of chitin, chitosan, and N-acetyl chitohexaose against Pseudomonas aeruginosa and Listeria monocytogenes infections in mice.

We conducted a comparative study of the protective effects of chitin, chitosan, and N-acetyl chitohexaose (NACOS-6) against mice infected intravenously or intraperitoneally with Pseudomonas aeruginosa and Listeria monocytogenes. Mice pretreated with chitin, chitosan, and NACOS-6 showed resistance to intraperitoneal infections by both microbes. Only mice pretreated with chitin and chitosan showed resistance to intravenous infections by both microbes. The number, active oxygen generation, and myeloperoxidase activity of peritoneal exudate cells (PEC) in the chitin, chitosan, and NACOS-6-treated mice were greater than those of the untreated mice. Also, these PEC factors from mice pretreated with chitin and chitosan were greater than those from the NACOS-6-treated mice.

Adjuvants, Immunologic↗
PubMed2003

Efficacy of methylprednisolone and urokinase pulse therapy for severe Henoch-Schönlein nephritis.

OBJECTIVE: To evaluate the efficacy of methylprednisolone and urokinase pulse therapy (MUPT) for severe Henoch-Schönlein nephritis, we examined the clinical manifestation and prognosis of patients with MUPT on long-term observation. METHODS: We enrolled 56 patients with Henoch-schönlein nephritis who had been diagnosed with at least type IIIb from 1980 to 1998 on long-term observation and had been treated with MUPT. The clinical features, laboratory data, and pathologic findings between "pre-MUPT" and "post-MUPT," and the prognosis of these patients on long-term observation were retrospectively investigated. RESULTS: The mean urinary protein excretion after 6 months of treatment had decreased significantly compared with "pre-MUPT." Hypercoagulant state in "after the completion of urokinase pulse therapy" improved compared with "pre-MUPT." First renal biopsies were performed in all patients and second biopsies were performed in 27 patients. The activity index decreased significantly from 4.1 +/- 1.9 at first biopsy to 2.5 +/- 1.7 at second biopsy, while the chronicity index did not differ between first and second biopsy. None had renal insufficiency and renal survival rate was 100% for the decade. CONCLUSIONS: Although uncontrolled, our study suggested that MUPT is effective for those patients with the risk of progression of their nephropathy, especially if started early during the course of the disease before the crescents become fibrous.

Child↗
PubMed2003

Oral disodium cromoglycate and ketotifen for a patient with eosinophilic gastroenteritis, food allergy and protein-losing enteropathy.

We present a case report of a 10 years old boy with protein-losing enteropathy and eosinophilic gastroenteritis who had positive histamine release tests, increased allergen-specific IgE antibodies to some food items, and low levels of total serum protein and albumin. Upper gastrointestinal endoscopy revealed a number of polyps and diffuse gastritis. Biopsy specimens of the stomach and duodenum showed widespread eosinophilia and neutrophilia. Although a restricted diet was recommended, a diet which excluded foods with positive results to both histamine release test and allergen-specific IgE antibodies was poorly tolerated, and the patient rejected systemic administration of corticosteroids. Thus, we initiated an oral disodium cromoglycate (DSCG) and ketotifen therapy. After oral DSCG and ketotifen administration, the patient's condition improved gradually. Therefore, oral DSCG and ketotifen therapy might be considered as treatment option in patients with eosinophilic gastroenteritis and protein-losing enteropathy caused by food allergy.

Administration, Oral↗
PubMed2002

Simultaneous estimation of the association constants of glycoprotein glycoforms to a common protein by capillary electrophoresis.

The efficacy of our capillary electrophoresis method for simultaneous estimation of the association constants of glycoprotein glycoforms to a common target protein was demonstrated using ribonuclease and ovalbumin glycoforms as glycoform models and Lens culinaris agglutinin (LCA) as a protein model. The ribonuclease glycoforms were fairly well separated in the absence of LCA at pH 5.8, but the peaks were retarded without any change of separation profile in the presence of LCA, the retardation becoming greater as LCA concentration increased. The estimated values of apparent association constant (K(a)) were at the 10(6)M(-1) level for all the ribonuclease glycoforms, and there was no significant difference among glycoforms. The high-mannose-type N-glycans released from a mixture of ribonuclease glycoforms gave lower values of K(a) at the 10(4)-10(5)M(-1) level to the same protein, and the glycans having a larger number of the mannose residue gave larger K(a) values. These results imply that the glycan moiety in this glycoprotein might contribute to its binding to the protein, but the polypeptide core played the major role. In contrast, ovalbumin glycoforms gave poorly resolved peaks in the absence of LCA, but they were separated into several peaks in the presence of LCA, which were tentatively assigned based on the knowledge of affinity to this lectin, and K(a) values were estimated simultaneously. The estimated K(a) values were smaller than those of the ribonuclease glycoforms, suggesting the major role of the N-glycan moiety. Thus, capillary electrophoresis allowed simultaneous estimation of K(a) values under common conditions using small amounts of glycoform mixtures and proteins without prior isolation and purification. Comparison of the obtained values will provide useful information on the glycan structure-affinity correlation.

Amino Acid Sequence↗
PubMed2002

Delayed lethal response to Aspergillus fumigatus infection in sarcoma 180 tumor-bearing mice.

A longer survival and a decrease in the number of fungal cells in kidneys and brain were observed in groups of mice inoculated with Aspergillus fumigatus conidia 2-3 weeks (especially 3 weeks) after sarcoma 180 tumor transplantation compared to groups of non-tumor-bearing (control) mice inoculated with fungal cells only. The 3-4-week tumor-bearing mice had significantly decreased levels of serum iron and increased levels of unbound iron binding capacity in the serum compared to those of the non-tumor-bearing mice.

Animals↗
PubMed2002

Colominic acid: a novel chiral selector for capillary electrophoresis of basic drugs.

We introduced colominic acid as a new chiral selector for capillary electrophoresis of basic drugs. Use of a low concentration phosphate buffer containing this polysaccharide and a Polybrene/colominic acid double coated capillary allowed excellent separation of the enantiomers of primaquine, chloroquine and tryptophan. Other drugs giving partial enantioseparation include laudanosine and salbutamol. Capillary coating with Polybrene followed by colominic acid eliminated the problems of peak tailing and low reproducibility of migration time in uncoated capillaries. The optimum pH was in the acidic region but varied among drugs. A low capillary temperature of 16 degrees C and a colominic acid concentration of 9 w/v% are recommended for practical analysis of these drugs. Colominic acid preparations having higher molecular masses gave better enantioseparation, and N-acetylneuraminic acid, the component monosaccharide, did not give any enantioseparation.

Electrophoresis, Capillary↗
PubMed2002

Estimation of sialic acid in a sialoglycan and a sialoglycoprotein by capillary electrophoresis with in-capillary sialidase digestion.

An example of application of in-capillary derivatization for CE, obtained by using the throughout-capillary format, is presented. Introduction of a sialoglycan (N-acetylneuraminyllactose) or a sialoglycoprotein (bovine serum fetuin) sample to a running buffer (pH 5.0) containing N-acetylneuraminidase followed by application of a voltage resulted in the release of N-acetylneuraminic acid (NANA) which could be estimated by CE with UV detection. Two-step application of voltages (5 and 20 kV) was proved to be more effective for rapid estimation of the released NANA. This format (modified throughout-capillary format) allowed differential estimation of the NANA present in the sample as an impurity and the NANA released from the substrate at the picomol level, and thereby reliable micro assay of the sialidase activity. It also allowed estimation of the rate constant of this enzymatic reaction.

Electrophoresis, Capillary↗
PubMed2002

Effect of structure modification of chondroitin sulfate C on its enantioselectivity to basic drugs in capillary electrophoresis.

The effect of structure modification of chondroitin sulfate C on its enantioselectivity to several representative basic drugs in capillary electrophoresis was investigated. Chemical desulfation showed no remarkable decrease in selectivity, whereas depolymerization with chondroitinase ABC resulted in complete loss of selectivity. Comparison with chondroitin sulfate A indicated considerable decrease in selectivity with this isomer. The great retention of enantioselectivity in the desulfated derivative suggests that the selectivity comes from the difference of the magnitude of an interaction in the multipoint mechanism between a part of the drug molecule and a functional group in chondroitin sulfate C other than the sulfate group. The sulfate group is not considered to play a major role for chiral separation. The complete loss of selectivity by depolymerization is consistent with a general tendency of lower selectivity in smaller saccharides, and the priority of chondroitin sulfate C to chondroitin sulfate A suggests the importance of the hydroxyl group at C4 in the galactosamine residue. During the course of this work we observed heavy tailing of the peaks of basic drugs in some batches of uncoated fused-silica capillaries under acidic conditions and solved this problem by doubly coating capillaries with Polybrene followed by chondroitin sulfate C. On the other hand, we demonstrated the usefulness of a special technique which uses a short, wider bore PTFE tube-attached capillary for the study of the effect of depolymerization, in order to minimize sample amount.

Carbohydrate Conformation↗
PubMed2002

Granulocyte colony-stimulating factor exacerbates the acute lung injury and pulmonary fibrosis induced by intratracheal administration of bleomycin in rats.

We investigated the effects of granulocyte colony-stimulating factor (G-CSF) on lung injury induced by intratracheal administration of bleomycin (BLM, 2 mg/200 micro1) in rats. In experiment 1, G-CSF (10, 30 and 100 microg/kg/day, s.c.) was administered to rats treated with BLM or saline for 7 days starting immediately after BLM administration. In rats receiving G-CSF alone, a large number of neutrophils were noted in the pulmonary capillaries, although there were no lung lesions. In rats receiving BLM alone, diffuse alveolar damage was observed. The administration of G-CSF to BLM-treated rats increased the total lung lesion per unit of pulmonary parenchyma (total lung lesion %) along with increases in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion in a dose-dependent fashion. In experiment 2, 100 microg/kg/day of G-CSF was administered to rats treated with BLM or saline for up to 28 days starting immediately after BLM administration. The administration of 100 microg/kg/day of G-CSF to BLM-treated rats showed no effects at 14 days but it increased the lung lesion % and the score of lung fibrosis along with the increase in the number of neutrophils infiltrating in the pulmonary lesion at 28 days. These findings suggest that G-CSF administration to BLM-treated rats influenced and exacerbated the BLM-induced acute lung injury, and also exacerbated pulmonary fibrosis in a dose-dependent fashion. The exacerbation of lung injury coincided with the marked increase in the peripheral neutrophil count and the number of neutrophils infiltrating in the pulmonary lesion.

Acute Disease↗
PubMed2002

Prediction of relapse by plasma lipoprotein(a) concentration in children with steroid-sensitive nephrotic syndrome.

AIM: To clarify whether plasma lipoproteins, including Lp(a), can predict relapse pattern in the first years after diagnosis of nephrotic syndrome (NS), we evaluated them in patients with steroid-sensitive NS. METHODS: We analyzed the medical records of 35 patients with steroid-sensitive NS who were seen by us from January 1992 to December 1999 followed for at least 1 year. These patients were divided into two groups. Group 1 consisted of 20 patients who infrequently relapse (IR: <2 in 6 months or <3 in a year), group 2 consisted of 15 patients who frequently relapse (FR: > or =2 in 6 months or > or =3 in a year). Clinical and laboratory findings such as age at onset, gender, urinalysis, serum level of total protein, albumin, and concentrations of serum lipid including lipoprotein(a) (Lp(a)) were investigated between group 1 and group 2. RESULTS: The concentration of plasma Lp(a) in group 2 was higher than that in group 1 (81.0 +/- 35.2 vs. 35.9 +/- 26.5 mg/dl, p < 0.01). On multivariate analysis using logistic regression model, the concentration of plasma Lp(a) was an independent risk factor for relapse of NS. CONCLUSIONS: Our findings suggest that of all the laboratory data examined, high values of Lp(a) can predict future relapse of NS and should be well documented.

Albumins↗
PubMedDate not supplied

Rapid and sensitive screening of N-glycans as 9-fluorenylmethyl derivatives by high-performance liquid chromatography: a method which can recover free oligosaccharides after analysis.

There are a large number of labeling methods for asparagine-type oligosaccharides with fluorogenic and chromophoric reagents. We have to choose the most appropriate labeling method based on the purposes such as mass spectrometry, high-performance liquid chromatography and capillary electrophoresis. Asparagine-type glycans are released from core proteins as N-glycosylamine at the initial step of the releasing reaction when glycoamidase F is employed as the enzyme. The N-glycosylamine-type oligosaccharides thus released by the enzyme are subjected to hydrolysis or mutarotation to form free-form oligosaccharides. In the detailed studies on the enzyme reaction, we found a condition in which the released N-glycosylamine-type oligosaccharides were exclusively present at least during the course of enzyme reaction, and developed a method for in situ derivatization of the glycosylamine-type oligosaccharides with 9-fluorenylmethyl chloroformate (Fmoc-Cl). The Fmoc labeled sialo- and asialo- (or high-mannose and hybrid) oligosaccharides were successfully analyzed on an amine-bonded polymer column and amide-silica column, respectively. The present method showed approximately 5 times higher sensitivities than that using 2-aminobenzoic acid (2-AA). The separation profile was similar to that observed using 2-AA method as examined by the analyses of carbohydrate chains derived from several glycoproteins including complex-type, high-mannose type and hybrid type of N-linked oligosaccharides. The labeled oligosaccharides were stable at least for several months when stored at -20 degrees C. Furthermore, it should be emphasized that the Fmoc-derivatized oligosaccharides could be easily recovered as free reducing oligosaccharides simply by incubation with morpholine in dimethylformamide solution. We obtained a pure triantennary oligosaccharide with 3 sialic acid residues as a free reducing form from fetuin in good yield after isolation of the corresponding Fmoc oligosaccharide followed by removing reaction of the Fmoc group. The proposed method will be useful for preparation of free oligosaccharides as standard samples at pmol-nmol scale from commercially available glycoproteins.

Asparagine↗