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Biomedical subjects

K Inagaki

Publications and source records attributed to K Inagaki.

At least 91 records · Page 5Linked to original sources
PubMed1994

Inhibition of tumor cell growth by low-boiling-point-saturated fatty acids isolated by molecular distillation and reversed phase liquid chromatography of hydrolysates of uncytotoxic wool grease secreted from sheep sebaceous gland.

We showed that massive growth of mouse Ehrlich ascites carcinoma (EAC) cells was not inhibited by wool grease secreted from the sheep sebaceous gland, whereas wool fatty acids separated by saponification of wool grease was growth-inhibitory. We then fractionated wool fatty acids into 9 fractions using molecular distillation (80-200 degrees C; 1 x 10(-2) mmHg) and found a marked antitumor activity in a low-boiling-point (< 80 degrees C) fraction (MW 200-300; C10-C20), which was further separated by reversed phase liquid chromatography on an octadecylisilica gel column, resulting in 5 fractions. The second most hydrophobic fraction (C8Si-4) obtained was the most growth-inhibitory to EAC cells cultured or implanted into mice, more marked than the antitumor glycopeptide bleomycin. C8Si-4 was suggested to be a mixture of a normal-chain C16-saturated fatty acid and two branched-chain kinds of saturated C16-iso- and C19-anteiso-fatty acids without hydroxyl groups according to gas chromatography-mass spectrographic analysis. Thus low-boiling-point saturated fatty acid moieties in some wool grease molecules were shown to become growth-inhibitory in vitro and in vivo only after released in the free acid form by esterolysis.

Animals↗
PubMed1993

Bacteriological investigation of bile in patients with cholelithiasis.

The microflora in bile from the gallbladder and common bile duct was investigated in 303 patients who underwent surgery for cholelithiasis. The purpose of this study was to identify current bacteria and bacterial casts in the biliary tract and also to analyze the relationship between bactericholia at the time of operation and postoperative infection. Bile cultures were positive in 38% of all patients, although a higher incidence of positive bile cultures occurred in patients over 70 years of age (77%), those with common duct stones (83%), those with pigment stones (65%), and those who underwent gastrectomy (71%). The predominant organisms were Escherichia coli (22%), Klebsiella (18%), and Enterococcus (15%). Obligate anaerobes were less frequently seen (4%), being found only in patients with pigment stones and always mixed with aerobes. Four patients developed postoperative infections (1.3%) which were all caused by biliary bacteria. The following two factors may contribute to this low incidence of postoperative infections: our policy of operating electively whenever possible, and the prophylactic use of antibiotics to which the organisms cultured from bile are sensitive.

Adult↗
PubMed1993

MEN type 1 associated with mediastinal carcinoid producing parathyroid hormone, calcitonin and chorionic gonadotropin.

A 34-year-old Japanese woman was admitted to our hospital with a mediastinal tumor. Laboratory data showed high levels of calcitonin (CT), human chorionic gonadotropin (hCG) and parathyroid hormone (PTH) in the serum. Surgery was performed, but the tumor could not be completely resected. The patient died in the 10th postoperative month due to systemic metastasis and cachexia. At autopsy, diffuse parathyroid hyperplasia and multiple islet cell tumors of the pancreas were found. A pathological diagnosis of multiple endocrine neoplasia (MEN) type 1 associated with mediastinal atypical carcinoid tumor was made. Immunohistochemical study indicated the tumor cells of the mediastinum to be positively stained for hCG beta but not for PTH or CT. The tumor tissue contained significantly high levels of CT, PTH and whole hCG. This is the first case report on MEN type 1 associated with mediastinal carcinoid tumor producing PTH, CT and hCG beta.

Adenoma, Islet Cell↗
PubMed1993

3-Isopropylmalate dehydrogenase from chemolithoautotroph Thiobacillus ferrooxidans: DNA sequence, enzyme purification, and characterization.

3-Isopropylmalate dehydrogenase encoded by the Thiobacillus ferrooxidans leuB gene was purified to homogeneity from Escherichia coli cells harboring a recombinant plasmid containing the leuB gene. The native enzyme molecule is a dimer of molecular weight 38,000. The Km value for 3-isopropylmalate was estimated to be 26 microM and that for NAD+ 0.8 mM. The presence of K+ or NH4+ is essential for the enzyme reaction. The enzyme is activated about 4-fold by the addition of 1.0 mM Mg2+ or Co2+. The optimum pH and temperature for the activity are 9.0 and 60 degrees C, respectively. The properties of the enzyme are similar to those of the Salmonella typhimurium and Thermus thermophilus enzymes, except for substrate specificity. T. ferrooxidans 3-isopropylmalate dehydrogenase is able to utilize D- and L-malate as substrates in addition to 3-isopropylmalate. Sequencing of subcloned DNA revealed that the leuB gene consists of a 1,074 bp open reading frame and encodes 358 amino acid residues corresponding to the subunit (38,462 Da). The amino acid sequence of 3-isopropylmalate dehydrogenase from T. ferrooxidans and those of some heterotrophic microorganisms have high homology.

3-Isopropylmalate Dehydrogenase↗
PubMed1993

Restriction endonuclease Aor13HI from Acidiphilium organovorum 13H, a new isoschizomer of BspMII: purification and characterization.

A restriction endonuclease, Aor13HI, an isoschizomer of BspMII, was purified to homogeneity from cell extracts of Acidiphilium organovorum strain 13H. The enzyme has a molecular mass of 60,000 daltons and consists of two subunits identical in molecular mass of 30,000 daltons. Aor13HI endonuclease, like BspMII, recognizes the palindromic six-base sequence 5'-TCCGGA-3', and cleaves between the T and C to produce a four-base 5' extension. Aor13HI is not inhibited by dam-dependent methylation. The isoelectric point of the enzyme is 5.7. Aor13HI activity was maximum at pH 7.5, 100 mM KCl, 7.5-10 mM MgCl2, and 55 degrees C. The enzyme was stable up to 60 degrees C. The N-terminal amino acid sequence (30 residues) of Aor13HI did not show any similarity with the sequence of other restriction endonucleases reported.

Amino Acid Sequence↗
PubMed1993

Transformation of the acidophilic heterotroph Acidiphilium facilis by electroporation.

We constructed a cloning vector for use in the acidophilic heterotroph Acidiphilium facilis. The vector pAH101 (8.8 kb) was constructed from a 6.1 kb restriction fragment of the Acidiphilium plasmid pAH1 and a pUC19 carrying a beta-lactamase gene. The antibiotic resistance gene was efficiently expressed in A. facilis. Several factors which influenced the transformation efficiency were optimized, resulting in a transformation efficiency of up to 3 x 10(3) transformants per microgram of plasmid DNA at a field strength of 10 kV/cm with a 7.0 ms pulse.

Electroporation↗
PubMed1993

Young children's understanding of the mind-body distinction.

A experiments investigated children's understanding of the mind-body distinction. Children of ages 4 and 5 recognized not only the differential modifiability of changeable versus unchangeable human properties and bodily versus mental properties, but also the independence of activities of bodily organs from a person's intention (Experiment 1). When presented 3 types of causal explanations (i.e., intentional, vitalistic, mechanical), 6-year-olds chose most often as most plausible for bodily functions vitalistic explanations (i.e., those ascribing the phenomena to a relevant bodily organ's initiative and effortful engagement in activity); 8-year-olds chose the vitalistic explanations second most often, following mechanical ones (Experiment 2). However, 6-year-olds, as well as 8-year-olds and adults, did not always choose vitalistic explanations over intentional explanations (Experiment 3); whereas they tended to prefer vitalistic explanations for biological phenomena, they predominantly accepted intentional ones for psychological phenomena (Experiment 3A). These results suggest that children as young as 6 years of age have acquired a form of biology as an autonomous domain which is separate from that of psychology.

Adolescent↗
PubMed1992

Cytosolic and intranuclear calcium signals in rat basophilic leukemia cells as revealed by a confocal fluorescence microscope.

A confocal fluorescence microscope with an argon-ion laser (488 nm) and a He-Cd laser (325 nm) was used to study spatial heterogeneity of the calcium signals in rat basophilic leukemia 2H3 cloned cell line (RBL-2H3). After stimulation with antigen (2,4-dinitrophenol-conjugated bovine serum albumin), fluo-3-fluorescence intensities increased in individual RBL-2H3 cells with different lag times. Time-dependent profiles of the fluo-3-fluorescence intensities resembled closely the patterns of the sequential fluorescence-ratio images of fura-2, which were used to measure the intracellular free-calcium concentration ([Ca2+]i) in individual RBL-2H3 cells using a conventional fluorescence microscope. The present results obtained using the confocal fluorescence microscope showed spatial heterogeneities of fluo-3-fluorescence intensities, suggesting the existence of spatial heterogeneity of [Ca2+]i in RBL-2H3 cells. That is, the results showed that calcium signals first occurred transiently at pseudopodia in RBL-2H3 cells, then the signals transferred to the central parts of the cells. In addition, from the fluorescence images of co-loaded Hoechst 33342 (bisbenzimide H 33342, a DNA-specific probe) which were produced by excitation with a He-Cd laser, it was found that the fluorescence images of the nucleus were quite similar to those of the calcium signals mentioned above. This suggested that the receptor-mediated calcium signals were transferred not only to the cytoplasm but also to the nucleus.

Animals↗
PubMed1992

A fluorescent molecular rotor probes the kinetic process of degranulation of mast cells.

A confocal fluorescence microscope was used to study the exocytotic secretory processes of mast cells in combination with an fluorescent molecular rotor, 9-(dicyanovinyl)julolidine (DCVJ). DCVJ is known to be an unique fluorescent dye which increases its quantum yield with decreasing intramolecular rotation. Here, DCVJ-loaded peritoneal rat mast cells were stimulated with compound 48/80 and their fluorescence images were compared with fluorescence calcium images of fluo-3-loaded mast cells. Subsequent to transient increases in intracellular free calcium ion concentration, DCVJ fluorescence increased dramatically in the cytoplasm and formed a ring-like structure around the nucleus, suggesting the possibility that the dye bound to the proteins composing the cytoskeletal architecture. Furthermore, the increases of DCVJ fluorescence intensities were mostly blocked in the presence of cytochalasin D (10 microM). However, fluo-3 fluorescence intensities still increased after addition of compound 48/80.

Aniline Compounds↗
PubMed1992

Conjunctival penetration of insulin and peptide drugs in the albino rabbit.

An in vitro model was used to evaluate the conjunctival penetration of three peptides, [D-ala2]metenkephalinamide (YAGFM, MW 647), substance P (MW 1348), and insulin (MW 5778), in comparison with two nonpeptides, atenolol (MW 266) and timolol (MW 433). All three peptides were hydrolyzed to varying extents during penetration across the conjunctiva. The permeability coefficient for intact YAGFM and insulin was 4.5 +/- 0.3 and 4.6 +/- 0.7 microns sec-1, respectively. These values were about two to five times lower than those for atenolol and timolol. No permeability coefficient could be calculated for substance P, since its transconjunctival flux never reached steady state. The conjunctival penetration of YAGFM and insulin was improved by about two and three times, respectively, with the addition of 1% Na glycocholate. Increasing the Na glycocholate concentration was more effective than changing the type of bile salt in improving the conjunctival penetration of insulin. The maximum factor of improvement was 12, as the Na glycocholate concentration was raised to 4%. The way in which Na deoxycholate, glycocholate, and taurocholate affected the conjunctival penetration of atenolol, timolol, and insulin suggests that these three bile salts improved mainly the transcellular penetration of the compounds studied.

Administration, Topical↗
PubMed1992

Pharmacokinetic population parameters for aminoglycosides in cholecystitis patients.

We report the use of a new method to determine patient population pharmacokinetic parameters (nonparametric expected maximum or NPEM). Our purpose was to develop and then analyze the utility of these parameters, compared to a more traditional approach. Nineteen patients with acute cholecystitis made up the control group for defining the parameters via NPEM. The standard of practice was to use a model created from a different intraabdominal infection group (appendicitis), referred to as "surgical patient model." These two models were compared with a group of 23 patients receiving gentamicin for acute cholecystitis. We concluded that the NPEM model was superior to the surgical patient model in predicting gentamicin trough and peak levels with less bias and better precision.

Adult↗
PubMed1992

Determination of fluconazole in human serum by solid-phase extraction and reversed-phase high-performance liquid chromatography.

A simple, rapid, and accurate reversed-phase octadecylsilyl high-performance liquid chromatographic method using solid-phase column extraction is described for measuring fluconazole in human serum. The column eluent was monitored by ultraviolet absorption at 210 nm. Fluconazole was extracted from diluted serum by adsorption on a small Bond-Elut C18 cartridge after the addition of UK48,134 as the internal standard and recovered by elution with methanol. The methanol was then evaporated to dryness and the residue reconstituted in 200 microliters of mobile phase and filtered prior to injecting an aliquot (50 microliters) onto an Adsorbosphere C18 column (4.6 x 250 mm, 5 microns particle size), using a mobile phase of 25 mM tris(hydroxymethyl)aminomethane-phosphate buffer (pH 7.0):acetonitrile (75:25, vol/vol). The retention times were 6.6 min for fluconazole and 9.0 min for the internal standard. The assay was precise, with inter- and intraassay coefficients of variation of less than or equal to 2.9% and less than or equal to 2.1%, respectively, and with good linearity (r = 1.000) in the range of 0.1 to 25 micrograms/ml. The duration of each analysis was 15 min and the minimum detectable serum concentration was 0.1 microgram/ml.

Administration, Oral↗
PubMed1992

Molybdenum oxidation by Thiobacillus ferrooxidans.

Thiobacillus ferrooxidans AP19-3 oxidized molybdenum blue (Mo) enzymatically. Molybdenum oxidase in the plasma membrane of this bacterium was purified ca. 77-fold compared with molybdenum oxidase in cell extract. A purified molybdenum oxidase showed characteristic absorption maxima due to reduced-type cytochrome oxidase at 438 and 595 nm but did not show absorption peaks specific for c-type cytochrome. The optimum pH of molybdenum oxidase was 5.5. The activity of molybdenum oxidase was completely inhibited by sodium cyanide (5 mM) or carbon monoxide, and an oxidized type of cytochrome oxidase in a purified molybdenum oxidase was reduced by molybdenum blue, indicating that cytochrome oxidase in the enzyme plays a crucial role in molybdenum blue oxidation.

Journal Article↗