Search PubMed⌕ Search

Biomedical subjects

H Kamiya

Publications and source records attributed to H Kamiya.

At least 145 records · Page 8Linked to original sources

Purification and characterization of Microcystis aeruginosa (freshwater cyanobacterium) lectin.

Microcystis aeruginosa, strain M228, a laboratory culture of freshwater cyanobacterium, showed hemagglutinating activity against rabbit, horse and human ABO erthrocytes. Crossed absorption tests revealed the presence of a single type of lectin in the extract of M228 strain cells. The lectin, termed MAL, was purified in combination with the affinity chromatography on acid-treated agarose gel and the gel permeation chromatography in an electrophoretically pure form. MAL was a glycoprotein containing 7.8% neutral sugars and was composed of a single polypeptide having a molecular weight of 57 kDa. Isoelectric point was estimated to be pH 6.4. Hemagglutinating activity of the lectin was inhibited effectively by N-acetyl-D-galactosamine and by glycoproteins. D-galactose and lactose also showed moderate inhibitory activity. The destruction of the hemagglutinating activity by a 2-mercaptoethanol treatment suggests the presence of intra-chain disulfide bond(s) essential for the activity in the molecule. The sequence of the amino-terminal region of MAL was determined as Val-Leu-Ala-Ser-Leu-Val-Ser-Thr-Ser-Gln-Ala-Gly-Ser-Leu-Glu-Leu-Leu- Ala [corrected].

ABO Blood-Group System↗

Ostrich intestinal glycohydrolases: distribution, purification and partial characterisation.

Intestinal glycohydrolases are enzymes involved in assimilating carbohydrate for nutrition. The avian forms of these enzymes, in particular the maltase-glucoamylase complex (MG), are not well characterised. This study encompassed characterisation of these enzymes from ostrich intestines, and the first kinetic analysis of an avian MG. Proteolytically solubilised MG from ileal brush border membrane vesicles was purified by Sephadex G-200 gel filtration and Tris-affinity-chromatography, while jejunal sucrase-isomaltase (SI) and MG were purified by Toyopearl-Q650 and phenyl-Sepharose chromatography. Amino acid sequences and compositions of enzyme subunits, resulting from SDS-PAGE, were determined. Kinetics of hydrolysis of linear oligosaccharides was studied. Ostrich MG and SI showed the highest activity in the jejunum, followed by the ileum and duodenum. No lactase or trehalase activity could be detected. The jejunal MG and SI, resulting from brush-border membrane vesicles, could not be separated during purification. However, a minor form of ileal MG was purified using Sephadex G-200 chromatography. Ileal MG contained three subunits of M(r) 145,000, 125,000 and 115,000. Although the N-terminal amino acid sequences bear no homology to SI, the M(r) 115,000 subunit shows homology to porcine MG in both sequence and amino acid composition. The pH optimum of maltose-, starch- and isomaltose-hydrolysing activity was 6.5 and that of sucrose-hydrolysing activity 5.5. The glycohydrolases were most active at 58 degrees C, but were quickly denatured above 60 degrees C. Sucrose- and starch-hydrolysing activities were more thermostable than maltose- and isomaltose-hydrolysing activities. Kinetic parameters (K(m), kcat and kcat/K(m)) for the hydrolysis of maltooligosaccharides, starch and glycogen are reported for ileal MG. Maltotriose and maltotetraose displayed partial inhibition of ileal MG. The study revealed large similarities between ostrich SI and MG in charge, size, shape and hydrophobicity, based on their inseparability by several methods. Measurement of the specificity constants for maltooligosaccharide hydrolysis by ileal MG revealed less efficient hydrolysis of longer substrates as compared to maltose and maltotriose.

Amino Acid Sequence↗

Pemphigus with features of both vulgaris and foliaceus variants, associated with antibodies to 160 and 130 kDa antigens.

We report an unusual variant of pemphigus in a 44-year-old man. He presented with scaly and crusted erosions associated with pruritic vesicles and erythema mainly on the chest, abdomen, back and face. Histology showed acantholysis with neutrophilic spongiosis in the granular layer and subcorneal region of the epidermis. Intercellular IgG in the epidermis was positive on direct immunofluorescence. Indirect immunofluorescence showed intercellular antibodies at a titre of 1 : 2 in the suprabasal epidermis. Circulating autoantibodies to 130 kDa and 160 kDa antigens were detected in the patient's serum by immunoblot analysis using epidermal extracts. These two antibodies eluted from individual nitrocellulose membranes reacted with the intercellular space in the epidermis on indirect immunofluorescence. This observation suggests that these antibodies correspond to desmogleins 3 and 1, respectively. The clinical symptoms almost completely disappeared after 28 days treatment with oral prednisolone (30 mg/day), leaving brown pigmented flecks on the lesional sites. These findings suggest that this patient's pemphigus has features of both the vulgaris and foliaceus variants, with antibodies against desmogleins 3 and 1.

Adult↗

Clinical utility of serum levels of eosinophil cationic protein (ECP) for monitoring and predicting clinical course in childhood asthma.

BACKGROUND: The concentration of ECP in serum has been proposed as a marker of airway inflammation in asthma. However, its clinical significance is still to be determined. OBJECTIVES: This study was performed to determine whether concentration of ECP in serum reflects clinical status in asthma and can serve as a predictive parameter. METHODS: Cross-sectional analysis was performed in 28 children with asthma. A total of 91 blood samples was obtained to determine levels of ECP in serum and eosinophil counts. Forced expiratory volume in 1 s was also determined at the time of the sampling. Data were analysed on the basis of asthma symptoms in the 4 weeks before and the 4 weeks after sampling. RESULTS: Serum levels of ECP were significantly lower in patients who had been asymptomatic for 3 or 4 weeks before sampling than in patients who had been symptomatic or asymptomatic for only 1 or 2 weeks. In the former group, serum levels of ECP were higher when patients became symptomatic after sampling than when they remained stable, a finding that suggests that serum levels of ECP may have a predictive value in certain situations. Although the concentration of ECP in serum was not proved to be predictive in the latter symptomatic group, the concentration of ECP was significantly lower when measured again 4 weeks later when the patients' symptoms had resolved. In contrast, levels of ECP were unchanged when patients remained symptomatic, a finding that suggests serum levels of ECP may reflect the clinical response to therapy. CONCLUSIONS: Serum ECP may be a useful marker for monitoring and predicting the clinical course in asthma.

Adolescent↗

Activation of lymphocytes by HHV-6 antigen in normal children and adults.

We investigated the immune reaction of lymphocytes in response to human herpes virus-6 (HHV-6) in normal children and adults. Cell proliferation was assayed by measurement of the incorporation of 5-bromo-2'-deoxyuridine (BrdU) by enzyme-linked immunosorbent assay (ELISA), and changes in expression of interleukin-2 receptors (IL-2Rs) (alpha-, beta3-, and gamma-chain) were assayed by flow cytometry. Incorporation of BrdU and expression of IL-2Rs (alpha-, beta-, and gamma-chain) in CD4+, CD8+, and CD45RO+ lymphocytes were increased when peripheral blood mononuclear cells (PBMC) from HHV-6 seropositive children aged 3 to 12 years and adults were cultured with HHV-6 antigen compared with control antigen. In contrast, cord blood mononuclear cells (CBMC) and PBMC from seronegative children did not show cell proliferation and changes in expression of IL-2Rs. In seropositive children less than 2 years of age, the magnitude of cell proliferation was low and IL-2Rs (alpha-, beta-, and gamma-chain) in CD8+ cells and IL-2Rs (alpha-chain) in CD45RO+ cells were increased. These data suggest that children below the age of 2 had immature lymphocytic response to HHV-6 antigen. Deletion of monocytes from PBMC and the addition of a mixture of anti-IL-2Rs (alpha-, beta-, and gamma-chain) antibodies reduced cell proliferation in response to HHV-6, suggesting the requirement of the presence of monocytes and expression of IL-2Rs.

Adult↗

Childhood bacterial meningitis in Japan.

Haemophilus influenzae type b (Hib) vaccines currently are not used in Japan, but interest in preventing H. influenzae disease by immunization has grown. We performed a retrospective survey for bacterial meningitis in 6 prefectures of Japan. Questionnaires requested the age, sex, clinical outcome and identity of the etiologic organism, if known, of all patients with meningitis younger than 16 years of age who were admitted during calendar year 1994. Of 876 hospitals within the 6 study prefectures, 363 (41.4%) returned a completed questionnaire. There were 1769 cases of meningitis reported, of which 160 (9%) were considered bacterial in origin. H. influenzae was the most common cause of bacterial meningitis, accounting for 68 cases (43%). Sixty-six cases (97%) of H. influenzae meningitis occurred in children 4 years of age or younger, and 27 (40%) occurred in children <1 year of age. Calculated incidence rates based on the population of children 4 years of age or less for each prefecture ranged from 3.4 to 9.9 cases per 100000 (mean, 4.7 cases/100000). H. influenzae is the most common cause of meningitis in Japan, and the estimated incidence rates from this study are very similar to those previously reported from Japan. More comprehensive, prospective surveillance studies will be needed to define better the incidence of Hib meningitis and to aid in making rational decisions regarding the use of Hib vaccination in Japan.

Adolescent↗

Accelerated influx of dendritic cells into the lymph nodes draining skin sites exposed to attenuated cercariae of Schistosoma mansoni in guinea-pigs.

We compared temporal changes in the cell composition of the skin-draining lymph nodes (SLN) in guinea-pigs exposed percutaneously to normal or attenuated cercariae of Schistosoma mansoni. Different populations were analysed by flow cytometry of double-stained cells by monoclonal antibodies to the major histocompatibility complex class II molecule and lymph node cells of guinea-pigs. Exposure to S. mansoni caused a marked increase in the proportion of B cells and dendritic cells (DC) on day 2, reaching a peak number on day 4. These changes were comparable in both infected and vaccinated animals exposed to normal or attenuated parasites, respectively. Total number of DC, however, showed different kinetics; in infected animals, the number peaked on day 2 and then gradually declined, whereas it reached a higher peak on day 4 in vaccinated animal. Daily injection of bromo-deoxyuridine after exposure to the parasite reduced the total number of DC in the SLN on day 4. A reduction in DC counts in the contralateral side SLN was also evident in vaccinated animals. Our results indicate that a significant number of newly formed DC are recruited to the skin by 4th day of vaccination, followed by increased efflux to the SLN. It is possible that retention of attenuated S. mansoni in the skin may cause accelerated recruitment of newly formed DC from the bone marrow, and facilitate transport and processing of antigens highly expressed on attenuated parasites.

Animals↗

Diagnosis of bacterial enteric infections in children in Zambia.

BACKGROUND: The bacterial pathogens commonly responsible for diarrhea in children under the age of 5 in Zambia were identified and the most effective methods of diagnosis of such infections in laboratories with limited resources, such as those in developing countries, are recommended. METHODS: Stool samples were collected from children under the age of 5 years who visited the Diarrhoea Training Unit (DTU) of Zambia University Teaching Hospital in Lusaka, Zambia, between May 1992 and May 1993. A total of 639 children were evaluated for the presence of bacterial infection using standard culture media. The prevalence of bacterial pathogens was compared with that reported from other developing countries. RESULTS: Pathogenic strains of Escherichia coli were isolated from 95 (14.9%) children, Shigella species from 65 (10.2%) children, and Vibrio cholerae from 21 (3.3%) children. The presence of visible blood in the feces was an early indicator of the presence of shigellosis. CONCLUSIONS: E. coli, Shigella species and Vibrio cholerae were the major causes of bacterial diarrhea in the Zambian children studied. Research is required to determine the prevalence of such enteropathogenic strains. The use of adequate diagnostic procedures is indispensable to appropriate management. The recommendations have been prepared as a manual for the identification of enteropathogenic bacteria to be used in laboratories with limited resources, such as in developing countries.

Bacteriological Techniques↗

Changes in adrenocorticotropic hormone (ACTH) release from the cultured anterior pituitary cells of streptozotocin-induced diabetic rats.

We examined ACTH release from cultured anterior pituitary cells of streptozotocin (STZ)-induced diabetic rats. Rats 1 week after the injection of STZ (55 mg/kg, i.v.) were used as a short-term diabetic model, while rats 8 weeks after the injection of STZ were used as a long-term diabetic model. ACTH release induced by corticotropin-releasing factors (CRF) was significantly increased in the short-term diabetic rats, whereas ACTH release decreased in the long-term diabetic rats. A stimulator of adenylate cyclase, forskolin, caused marked increases in ACTH release in short-term diabetic rats, whereas it did not affect the long-term diabetic rats. An L-type Ca2+ channel agonist, BAY K 8644, did not affect ACTH release in the short-term diabetic rats, although it decreased ACTH release in long-term diabetic rats. In addition, CRF-induced ACTH release also increased in normal anterior pituitary cells cultured under high glucose conditions (25 mmol/L) for 5 d. These results suggest that the increase in the CRF-induced ACTH release from cultured anterior pituitary cells of short-term diabetic rats appears to involve the cAMP system in part. In contrast, the decrease in the CRF-induced ACTH release from the cultured anterior pituitary cells of long-term diabetic rats may indicate a change in the properties of the L-type Ca2+ channel coupled with the CRF receptor, or in the CRF receptor itself.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

[Effects of arginine-vasopressin on neuronal interaction from the area postrema to the nucleus tractus solitarii in rat brain slices].

The effects of vasopressin (AVP) on area postrema (AP) neurons and the neuronal connection between the AP and nucleus tractus solitarii (NTS) were investigated electrophysiologically in slices preparation of the medulla oblongata of rats. In the AP, 27.9% of 129 neurons were excited by AVP and 20.5% were inhibited. The excitation was blocked by a V1 receptor antagonist. Synaptic transmission of the AP to the NTS was mainly mediated by non-NMDA receptors. Local application of AVP to the AP activated the NTS neurons. This activation was blocked by an NMDA antagonist. These results suggest that the excitation originating in the AP is conveyed to the NTS via non-NMDA receptors and modified by NMDA receptor activation secondly. These processes may be important in regulation of the arterial baroreceptor reflex.

Animals↗

Immunization of laboratory animals with ultraviolet-attenuated larvae against homologous challenge infection with Trichinella britovi.

Immunogenicity of Trichinella britovi (Japanese isolate, cord ISS 408) muscle larvae irradiated with 5 mJ/cm2 of short wavelength ultraviolet (UV) was studied using homologous challenge infections. SCID mice vaccinated with UV-attenuated larvae showed no resistance, although its congenic CB-17 mice exhibited remarkable immunity. Following challenge vaccinated Mongolian gerbils, Meriones unguiculatus, did not show significant immunity. Use of these animal models vaccinated with UV-attenuated larvae will contribute to the investigation of immune mechanisms against Trichinella infection and also to the development of a vaccine of trichinellosis.

Animals↗

Schistosoma mansoni infection in IgE-producing and IgE-deficient mice.

The immunoglobulin E (IgE) response, a hallmark of helminthic infection, is generally considered a major host defense against schistosomiasis mansoni. In support, it was reported that mice with a null mutation of the Ce gene, which are thus incapable of making IgE, developed Schistosoma mansoni worm burdens 2-fold greater than wild-type mice. However, in another study, reduction of the IgE response in mice to a primary S. mansoni infection by anti-IgE treatment resulted in decreased worm burden and fecundity, suggesting that IgE plays a detrimental, rather than beneficial, role for the host in schistosomiasis. In a third study, S. mansoni worm burden and egg production in normal and in IL-4-deficient mice that produce negligible IgE levels did not differ significantly, and it appeared that IgE did not affect parasite survival or fecundity. In an attempt to resolve these controversies, we examined hepatic worm load and egg production in the liver and small intestine of IgE-deficient (SJA/9) and control IgE-producing (SJL/J) mice, 8 wk after S. mansoni infection. No differences were observed in worm burden, total egg production, and number of eggs produced per female worm in the 2 mouse strains, confirming the data that imply that IgE does not play an essential role in primary S. mansoni infection.

Animals↗

[Duplex scanning of the internal thoracic artery for coronary artery surgery].

Ultrasonic duplex scanning was used to examine 77 internal thoracic arteries (ITA). The investigated vessels were classified as normal, stenosed (peak systolic velocity was faster than 150 cm/sec and doppler signals showed turbulent pattern), small (diameter measurements were smaller than 1.4 mm and mean flow velocities were slower than 20 cm/sec in distal part of the vessels), and occluded (no flow characteristics could be detected) by the doppler spectrum analysis and the two-dimensional B-mode images. The results of the duplex examination were compared with operative findings and angiograpic findings. Diameter measurements by duplex scanning compared with operative findings showed virtually no differences. One stenosed ITA, a 75% stenosis in the proximal part of the vessel, and two small ITAs were detected by duplex scanning. We performed coronary artery bypass grafting using distal part of the stenosed ITA as a free graft, and used small ITAs as individual bypass grafts. Two normal ITAs were misjudged as small, and one small ITA was misjudged as occluded. All normal ITAs of duplex examination showed normal findings of angiography. Duplex scanning is a reliable, sensitive, and noninvasive technique for the preoperative assessment of the ITA, and is a suitable screening for coronary artery surgery.

Coronary Artery Bypass↗

[Fundamental and clinical studies on tazobactam/piperacillin in the pediatric field].

Pharmacokinetics, efficacy and safety of tazobactam/piperacillin (TAZ/PIPC), a new beta-lactamase inhibitor combined with a penicillin antibiotic, were studied in pediatrics and the following results were obtained. 1) Serum concentration and urinary excretion of TAZ/PIPC after intravenous administration at a dose of 46 mg/kg to one child were investigated. Serum half-lives were 0.5 hours for TAZ and 0.6 hours for PIPC, while the urinary recovery rates in the first 6 hours after administration, were 61.6% and 56.3% respectively. 2) TAZ/PIPC was administered at a dose of 46.0-73.5 mg/kg t.i.d. for 3-13 days to 10 patients, 6 with respiratory infection, 1 with pharyngotonsillitis, 1 with pertussis and 2 with soft tissue infection of skin. Clinical response were excellent in 5 cases and good in 5 cases, and the efficacy rate was 100%. Bacteriologically, 2 strains (Staphylococcus aureus and Streptococcus pyogenes) isolated from 2 patients were eradicated after the treatment. As for adverse reaction, mild skin rash was observed in 1 case but disappeared within 2 days after withdrawal of the drug. As for abnormal laboratory test results decrease in neutrophiles and increase in eosinophiles in each 1 case were observed, which were, however, normalized after administration. From the above results, tazobactam/piperacillin was considered to be a useful antibiotic in the pediatric field.

Bacterial Infections↗

Mutations induced by 2-hydroxyadenine on a shuttle vector during leading and lagging strand syntheses in mammalian cells.

An oxidatively damaged base, 2-hydroxyadenine (2-OH-Ade), was incorporated into a predetermined site of one of the strands {(+)- or (-)-strand} of the double-stranded shuttle vector, pSVK3, and the modified DNAs were transfected into simian COS-7 cells. The nucleotide sequences in which the modified base was incorporated were 5'-GTCGA*C and 5'-CTTA*AG (A* represents 2-OH-Ade). The former is the recognition site for the restriction enzyme SalI, and the latter is that for AflII. The DNAs replicated in the cells were recovered and were transfected again into Escherichia coli. The DNAs recovered from the COS-7 cells transfected with a plasmid containing 2-OH-Ade at either site of the (+)-strand (a template strand for lagging strand synthesis) formed colonies about 50%-70% as frequently as the unmodified DNA. This indicated that the base weakly blocked DNA replication during lagging strand synthesis. On the other hand, the base in the (-)-strand did not appear to affect the efficiency of leading strand synthesis in COS-7 cells. The mutation frequencies of 2-OH-Ade in COS-7 cells were 0.6%-0.1%, depending on the sequence and the strand location. Although the mutation spectra of 2-OH-Ade also differed with sequences and strands, the base elicited substitution and deletion mutations in mammalian cells, as in E. coli. These results indicate that 2-OH-Ade is mutagenic in eukaryotic cells as well as in prokaryotic cells.

Animals↗

Mutational specificity of glyoxal, a product of DNA oxidation, in the lacI gene of wild-type Escherichia coli W3110.

To determine the mutation spectrum of glyoxal, which is produced from DNA by oxygen free-radicals, we analyzed the chromosomal lacI gene of mutants induced by treatment of a wild-type Escherichia coli strain with glyoxal. The cell death and the mutation frequency increased according to the concentration of glyoxal added to the culture medium. The majority of the spontaneous mutations (82%) and that of the glyoxal-induced mutations (50%) were the addition or deletion of a 5'-TGGC-3' sequence at positions 623-634, which was reported to be a mutational hot spot in the lacI gene. In the glyoxal-induced mutants, however, the ratio of base-pair substitutions was increased (35%). Although all types of base-pair substitutions were detected, 78% of the base-pair substitutions occurred at G:C sites. Among them, G:C-->A:T transitions were predominant, followed by G:C-->T:A transversions. These mutations appeared to be distributed randomly within the lacI gene. These results suggest that glyoxal-induced mutations may correlate to mutations induced by oxygen free-radicals.

Bacterial Proteins↗

Glyoxal, a major product of DNA oxidation, induces mutations at G:C sites on a shuttle vector plasmid replicated in mammalian cells.

Glyoxal is a major product of DNA oxidation in which Fenton-type oxygen free radical-forming systems are involved. To determine the mutation spectrum of glyoxal in mammalian cells and to compare the spectrum with those observed in other experimental systems, we analyzed mutations in a bacterial suppressor tRNA gene (supF) in the shuttle vector plasmid pMY189. We treated pMY189 with glyoxal and immediately transfected it into simian COS-7 cells. The cytotoxicity and mutation frequency increased according to the dose of glyoxal. The majority of glyoxal-induced mutations (48%) were single-base substitutions. Eighty three percent of the single-base substitutions occurred at G:C base pairs. Among them, G:C-->T:A transversions were predominant, followed by G:C-->C:G transversions and G:C-->A:T transitions. A:T-->T:A transversions were also observed. Mutational hotspots within the supF gene were detected. These results suggest that glyoxal may play an important role in mutagenesis induced by oxygen free radicals.

Animals↗