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Biomedical subjects

K Shiomi

Publications and source records attributed to K Shiomi.

At least 109 records · Page 6Linked to original sources

Establishment of radioimmunoassay for human neutrophil peptides and their increases in plasma and neutrophil in infection.

Neutrophil peptide, also called defensin, is an antimicrobial peptide localized in azurophil granules in neutrophils. Using a synthetic human neutrophil peptide-1 (HNP-1), we prepared an antiserum against the peptide and established a highly sensitive radioimmunoassay (RIA). This RIA equally recognized HNP-1, -2, and -3 on a molar basis. Analyses of leukocyte and blood extracts obtained from normal individuals and patients with bacterial infection by reverse-phase high-performance liquid chromatography coupled with the RIA showed that the antiserum specifically detected HNP molecules. Concentrations of HNP 1-3 in total in normal human blood and plasma are 13.2 +/- 1.4 (mean +/- SEM) ng/mu 1 and 403.8 +/- 38.9 pg/mu 1, respectively. The values increased in infection and returned to within the normal levels after the recovery of the disorder. Granulocyte colony-stimulating factor (G-CSF) increased the plasma and blood concentrations of HNP 1-3. The RIA established here provides a useful tool to elucidate the pathophysiological significance of HNP and to evaluate the neutrophil functions in infection and inflammation.

Acute Disease↗

A basic transthyretin variant (Glu61-->Lys) causes familial amyloidotic polyneuropathy: protein and DNA sequencing and PCR-induced mutation restriction analysis.

A new mutation of transthyretin (TTR) has been identified in a patient with late-onset familial amyloidotic polyneuropathy (FAP) of Japanese origin. Peptide mapping by reverse-phase high performance liquid chromatography to compare the patient's TTR with normal TTR showed the presence of an abnormal peptide. Amino acid sequence analysis of the peptide (residues 49-61) showed a lysine-for-glutamic acid substitution at position 61. This substitution, verified by direct DNA sequencing, was the result of a guanine to adenine change on exon 3 of the TTR gene. A polymerase chain reaction-induced mutation restriction analysis (PCR-IMRA) system was established to rapidly detect the missense mutation. TTR-Lys61 is the first variant TTR with a replacement of the acidic with basic amino acid to be found in the amyloid precursor proteins of FAP.

Aged↗

Differences of intravenous nitroglycerin responses in left ventricular systolic and end-diastolic pressures and coronary artery diameters during long-term treatment with cutaneous nitroglycerin patches.

The differences of intravenous nitroglycerin responses in left ventricular (LV) systolic and end-diastolic pressures and in coronary artery diameters (cross-tolerance) were investigated in patients receiving nitroglycerin patches. During diagnostic cardiac catheterization, graded doses of 50, 100, and 150 mcg of intravenous nitroglycerin were given. Left ventricular systolic and end-diastolic pressures and left coronary arteriograms were obtained during each dose. Twenty patients with coronary artery disease were studied. Before cardiac catheterization, 10 received nitroglycerin patch (patch group), and 10 did not (control group). In the control group, graded intravenous nitroglycerin doses of 50, 100, and 150 mcg caused decrease in LV systolic pressure of 18 +/- 7%, 20 +/- 5%, and 23 +/- 6%, respectively. In the patch group, the same intravenous nitroglycerin doses decreased LV systolic pressure by 12 +/- 6% (p < 0.05), 19 +/- 7% (NS), and 18 +/- 6% (p < 0.05), respectively, (p value: vs. control group). At the same intravenous nitroglycerin doses, LV end-diastolic pressures were decreased by 48 +/- 14%, 52 +/- 17%, and 56 +/- 9%, respectively, in the control group. However, there were no significant differences in LV end-diastolic pressure between the two groups for any of the three intravenous nitroglycerin doses. The same intravenous nitroglycerin doses caused increase in diameter of the left anterior descending coronary artery and circumflex coronary artery in the control group, which was attenuated significantly in the patch group. Tolerance may develop in LV systolic pressure and coronary artery diameters, whereas it may not develop in LV end-diastolic pressure.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Cutaneous↗

Affinity chromatographic purification of tetrodotoxin by use of tetrodotoxin-binding high molecular weight substances in the body fluid of shore crab (Hemigrapsus sanguineus) as ligands.

A purification method for tetrodotoxin (TTX), based on affinity chromatography using the TTX-binding high mol. wt substances in the body fluid of shore crab (Hemigrapsus sanguineus) as ligands, was developed. This method was particularly useful for analysis of TTX in biological samples with low concentrations of TTX. The affinity gel prepared was highly specific for TTX, having no ability to bind 4-epi-TTX and anhydro-TTX as well as saxitoxin.

Ammonium Sulfate↗

Zollinger-Ellison syndrome and pheochromocytoma. Report of a case.

A 49-year-old woman was diagnosed in 1985 as having pheochromocytoma because of hypertension with high levels of plasma catecholamine concentration and 24-hour urine excretion of vanillyl-mandelic acid and metanephrine together with a right adrenal mass. The excised tumor cells had fine granular basophilic cytoplasm with argyrophilic granules by Grimelius' method. Four years later, she was diagnosed as having a duodenal bulb ulcer. Serum gastrin showed an abnormally high level of 1900 pg/ml. Abdominal echogram and computed tomography revealed a hypoechoic lesion in the pancreas and intrahepatic multiple tumors. A needle biopsy specimen of the liver tumor was compatible with the histology of metastatic islet cell tumor. A diagnosis of Zollinger-Ellison syndrome was made due to malignant gastrinoma with multiple liver metastases. The patient had no family history of endocrinological or neoplastic disorders. The present case indicates the possibility that pheochromocytoma and gastrinoma, that is, endocrine tumors characteristic of multiple endocrine neoplasia (MEN) I and MEN II, may be coincident even in a person without MEN. A continued awareness of previously rare or undescribed manifestations is important in patients with islet cell tumors or pheochromocytoma.

Adrenal Gland Neoplasms↗

Identification of a Drosophila homologue of alpha-catenin and its association with the armadillo protein.

The cadherin cell adhesion system plays a central role in cell-cell adhesion in vertebrates, but its homologues are not identified in the invertebrate. alpha-Catenins are a group of proteins associated with cadherins, and this association is crucial for the cadherins' function. Here, we report the cloning of a Drosophila alpha-catenin gene by low stringent hybridization with a mouse alpha E-catenin probe. Isolated cDNAs encoded a 110-kD protein with 60% identity to mouse alpha E-catenin, and this protein was termed D alpha-catenin. The gene of this protein was located at the chromosome band 80B. Immunostaining analysis using a mAb to D alpha-catenin revealed that it was localized to cell-cell contact sites, expressed throughout development and present in a wide variety of tissues. When this protein was immunoprecipitated from detergent extracts of Drosophila embryos or cell lines, several proteins co-precipitated. These included the armadillo product which was known to be a Drosophila homologue of beta-catenin, another cadherin-associated protein in vertebrates, and a 150-kD glycoprotein. These results strongly suggest that Drosophila has a cell adhesion machinery homologous to the vertebrate cadherin-catenin system.

Amino Acid Sequence↗

Mutation of twins encoding a regulator of protein phosphatase 2A leads to pattern duplication in Drosophila imaginal discs.

The Drosophila gene twins was identified through a P-element-induced mutation that caused overgrowth in posterior regions of the wing imaginal disc. Analyses using position-specific markers showed that the inactivation of this locus induced the formation of extra wing blade anlagen in the posterior compartment of the disc. The duplication was mirror symmetrical, and the line of the symmetry did not correspond to any of the known compartment borders. We isolated the twins gene and found that it encoded one of the regulatory subunits of protein phosphatase 2A (PP2A). These results suggest a novel aspect of physiological roles of protein dephosphorylation; that is, the control of PP2A activity is crucial for specification of tissue patterns.

Amino Acid Sequence↗

Autooxidation of alkylhydrazones and mutagenicity of the resulting hydroperoxides.

Acetone alkylhydrazones were readily autooxidized to 2-alkylazo-2-propyl hydroperoxides, which were directly mutagenic in Salmonella typhimurium TA1535, TA100, TA102 and Escherichia coli WP2hcr. The mechanism of this mutagenicity presumes that the hydroperoxides in aqueous solution decompose to alkyl diazonium ions which were observed in the alkylation of 4-(p-nitrobenzyl)pyridine, and also to hydroxyl radical which was detected by ESR.

Alkylation↗

A new antibiotic, cypemycin. Taxonomy, fermentation, isolation and biological characteristics.

A new peptide antibiotic, cypemycin, with a molecular weight of 2,097 (M+H), was isolated from the culture broth of Streptomyces sp. OH-4156. The antibiotic possesses cytocidal activity against P388 leukemia cells in vitro at a concentration of 1.3 microgram/ml (IC50 values), and the antibiotic showed antimicrobial activities against Micrococcus luteus (MIC, 0.2 microgram/ml).

Amino Acid Sequence↗

Establishment of hypersensitive radioimmunoassay for islet amyloid polypeptide using antiserum specific for its N-terminal region.

Using a synthetic N-terminal hexadecapeptide of islet amyloid polypeptide (IAPP), we prepared an antiserum specific for IAPP[1-16] and established an extremely sensitive radioimmunoassay (RIA) for the peptide with a minimum detection level of 0.26 fmol/tube. Since the N-terminal sequence of IAPP is 100% conserved in many mammalian species, the RIA is widely applicable in quantifying their IAPP. Analyses of pancreatic extracts of human and hamster using reverse-phase high performance liquid chromatography coupled with the RIA revealed that almost all pancreatic IAPP consisted of IAPP[1-37]. On the other hand, rat and mouse pancreata contained substantial amounts of IAPP[1-16] and IAPP[1-17] in addition to IAPP[1-37] as a major molecular form. In human plasma, IAPP[1-37] is the major molecular form secreted into the circulation in response to glucose administration. The RIA established in this study is promising in elucidating the physiological functions and the pathophysiological significance of IAPP in diabetes mellitus.

Adult↗

Identification of a novel transthyretin variant (Val30----Leu) associated with familial amyloidotic polyneuropathy.

A novel variant transthyretin which contains a leucine-for-valine substitution at position 30 was isolated and identified in the serum of a patient with familial amyloidotic polyneuropathy (FAP). The amino acid substitution was proven to result from a guanine-to-cytosine change at the first base of codon 30 located in exon 2 in the mutated transthyretin gene by restriction fragment length analysis on the amplified transthyretin gene using Cfr13 I. The study indicates that the point mutation of the transthyretin gene is a cause of the disorder.

Amino Acid Sequence↗

Metabolic bone disease following gastrectomy: assessment by dual energy X-ray absorptiometry.

Although disorders in bone metabolism have long been recognized as typical sequelae of gastrectomy, there has until now been no reliable method of providing precise assessment of bone mass, resulting in a variation of reported incidence. In this study, metabolic bone disease was evaluated in a well characterized population: 34 men 2-5 years after gastrectomy; 11 men 6-10 years after gastrectomy; eight men 2-5 years after colonic resection; and 115 healthy men. The age range was 50-69 years. An innovative reliable method of dual energy X-ray absorptiometry was used which allows quantitative assessment of bone mineral content. Measurement of lumbar spine bone mineral density revealed that the mean(s.e.m.) bone mineral density in patients who had had a gastrectomy (2-5 years 0.84(0.03) g/cm2, 6-10 years 0.85(0.05) g/cm2) was significantly lower than that of patients who had undergone colonic resection (0.96(0.04) g/cm2) or healthy men (0.96(0.03) g/cm2). This study demonstrates that a high incidence of bone atrophy is induced at the relatively early period of 2-5 years after gastrectomy.

Absorptiometry, Photon↗

Islet amyloid polypeptide plasma concentrations in individuals at increased risk of developing type 2 (non-insulin-dependent) diabetes mellitus.

To study whether abnormal secretion of islet amyloid polypeptide is involved in the development of insulin resistance and impaired insulin secretion in Type 2 (non-insulin-dependent) diabetes mellitus, we measured islet amyloid polypeptide concentrations in 56 first-degree relatives of Type 2 diabetic subjects and in 10 healthy control subjects. Fasting islet amyloid polypeptide concentrations were similar in control subjects, glucose-tolerant and glucose-intolerant relatives (8 +/- 1, 9 +/- 1 and 11 +/- 2 fmol/ml; p = NS). The area under the islet amyloid polypeptide curve measured during an oral glucose load was larger in glucose-intolerant relatives (115 +/- 13 fmol/ml) compared to glucose tolerant relatives and control subjects (88 +/- 3 and 79 +/- 12 fmol/ml; p less than 0.05). The insulin response during the oral glucose load was inversely correlated with the rate of glucose disposal measured during a euglycaemic hyperinsulinaemic clamp (r = -0.725; p less than 0.01), while no significant correlation was observed between the corresponding values for islet amyloid polypeptide and glucose disposal (r = -0.380; p = NS). Hypersecretion of islet amyloid polypeptide is observed in glucose-intolerant first-degree relatives of patients with Type 2 diabetes. Since these patients are characterized by insulin resistance and abnormal first-phase insulin secretion, the putative role of islet amyloid polypeptide in the development of these abnormalities remains to be established. It is however, unlikely that islet amyloid polypeptide is involved in the development of insulin resistance as insulin-resistant relatives with normal glucose-tolerance showed normal islet amyloid polypeptide concentrations.

Adult↗

Occurrence of tetrodotoxin-binding high molecular weight substances in the body fluid of shore crab (Hemigrapsus sanguineus).

The shore crab (Hemigrapsus sanguineus) is highly resistant to tetrodotoxin (TTX) although it contains no detectable amount of TTX (less than 5 MU/g, where 1 MU is defined as the amount of TTX killing a 20 g mouse in 30 min). Its body fluid was examined for neutralizing effects against the lethal activity of TTX. When the mixture of the body fluid and TTX was injected i.p. into mice, the lethal activity of TTX was significantly reduced; 1 ml of the body fluid was evaluated to neutralize 3.6-4.0 MU of TTX. Higher neutralizing activity (7.2-12.5 MU/ml of the body fluid) was exhibited by i.v. administration of the body fluid into mice before or after i.p. challenge of TTX. The lethal effect of paralytic shellfish poisons was not counteracted by the body fluid. Analysis by gel filtration on Sepharose 6B revealed that the body fluid contained TTX-binding high mol. wt substances (> 2,000,000) responsible for the neutralizing activity of the body fluid against TTX, which accounts for the high resistibility of the crab to TTX. When the crude toxin extracted from the liver of puffer (Takifugu niphobles) was mixed with the body fluid and chromatographed on Sepharose 6B, almost pure TTX was obtained from the fractions containing the TTX-binding high mol. wt substances, suggesting that the TTX-binding high mol. wt substances could be useful in purification of TTX from biological samples.

Animals↗

Molecular forms of islet amyloid polypeptide (IAPP/amylin) in four mammals.

Using reverse-phase high performance liquid chromatography combined with radioimmunoassays for human and rat/mouse islet amyloid polypeptide (IAPP), we identified molecular forms of IAPPs in pancreata of four mammals including species in which islet amyloid deposition occurs (human and cat) and those in which amyloid deposition does not occur (rat and mouse). In human pancreas, IAPP (1-37) was the major molecular form, and IAPP (17-37), IAPP (24-37) and four IAPP-immunoreactive peptides were detected as minor components. In rat, mouse and cat pancreata, IAPP (1-37) and IAPP (19-37) were identified with the latter being the major molecular form. Major processing takes place at a single arginine residue at position 18 of rat/mouse and cat IAPPs, but not at the histidine at position 18 of human IAPP, indicating that arginine could yield different processing of IAPP between the 3 species and human. Different processing of IAPP by species suggests that processing of IAPP in pancreas is not responsible for islet amyloid formation. Identification of molecular forms of IAPP is helpful in elucidating the physiological function of the IAPP molecule and in determining the type of system regulating biosynthesis and catabolism of the peptide.

Amino Acid Sequence↗

Clinical features of 125 patients with sarcoidosis: Okayama University Hospital review of a recent 10-year period.

Clinical features were studied in 125 patients with sarcoidosis (72 females and 53 males) diagnosed at Okayama University Hospital during a recent 10-year period. The age distribution had two peaks in patients in their 20s and the 50s. Over half of the patients were detected at health screening check and were asymptomatic, while the remaining were symptomatic. Twelve patients were in stage 0, 41 were in stage I, 54 were in stage II, 16 were in stage III, and 2 were in stage IV according to the chest x-ray findings. Serum angiotensin converting enzyme levels and serum lysozyme levels were elevated in 60% and 76% of the patients, respectively. The bronchoalveolar lavage fluid showed lymphocytosis, especially of helper T-cells. The clinical features of sarcoidosis appear to depend on the duration of the disease.

Adolescent↗