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Biomedical subjects

K Matsuda

Publications and source records attributed to K Matsuda.

At least 433 records · Page 24Linked to original sources

Glycosphingolipid compositions of human T-lymphotropic virus type I (HTLV-I) and human immunodeficiency virus (HIV)-infected cell lines.

Glycolipid compositions of cells infected by human retroviruses (human immunodeficiency virus, HIV and/or human T-cell lymphotropic virus type I, HTLV-I) have been studied. Eight cell lines, comprising two HTLV-I-infected T-cell lines (MT-2 and MT-4), two HTLV-I-negative T-cell lines (Jurkat and MF), a macrophage cell line (U937), and three HIV-infected counterpart cell lines (MT-4/HIV, Jurkat/HIV and U937/HIV) were used. The neutral glycolipids and gangliosides isolated from these cell lines were compared. Among them, the HTLV-I-infected T-cell lines, MT-2 and MT-4, showed similar patterns for both neutral glycolipids and gangliosides. Neutral glycolipids (GlcCer and LacCer) of MT-2 and MT-4 cells were markedly decreased, and a ganglioside, GM3, of theirs was decreased to only a trace amount compared to that in other cell lines. Gangliosides of MT-4 and MT-4/HIV were further separated on an Iatrobeads column, and were identified as GM2, GM1a and GD1a by methylation and liquid secondary ion mass spectrometric analyses. Since the patterns of neutral glycolipids and gangliosides of MT-2 and MT-4 are unique, as compared to those of HTLV-I-negative cells, it is suggested that these changes are related to HTLV-1 infection. No prominent differences in the ganglioside compositions between HIV-infected and non-infected cell lines could be observed. But it is noteworthy that the contents of asialo-GM2 in Jurkat/HIV and MT-4/HIV cells were increased as compared to those in the parental cell lines.

Carbohydrate Sequence↗

Phorbol ester stimulates the activity of a protein tyrosine phosphatase containing SH2 domains (PTP1C) in HL-60 leukemia cells by increasing gene expression.

The affinity-purified antibody to a protein tyrosine phosphatase (PTP) containing two src homology 2 domains (PTP1C) was generated. The antibody recognized two types of PTP1C (PTP1C-alpha and -beta) of which the molecular sizes were 66 (alpha) and 62 kDa (beta), respectively, and these two types were expressed differentially in various cell types. The immune complex phosphatase assay using the antibody demonstrated that 12-O-tetradecanoylphorbol-13-acetate (TPA) and a vitamin D metabolite increased the PTP activity of immunoprecipitated PTP1C to 230 and 150% of control, respectively. By contrast, neither dimethyl sulfoxide nor retinoic acid significantly affected the PTP activity of PTP1C in HL-60 cells. The time course increment by TPA of PTP1C activity was closely correlated with that of the acquisition by HL-60 cells of a macrophage-like phenotype. In addition, TPA increased the amount of PTP1C detected by immunoblotting and immunoprecipitation and raised the level of expression of PTP1C mRNA in HL-60 cells. The increase of PTP1C mRNA induced by TPA treatment was inhibited by cycloheximide, suggesting that new protein synthesis is required for the increase by TPA of PTP1C mRNA expression. Furthermore, TPA increased the rate of transcription of the PTP1C gene without affecting the stability of PTP1C mRNA. These results suggest that (i) two subtypes of PTP1C may exist and function in various cell types, and (ii) TPA stimulates the PTP activity of PTP1C by increasing the transcription rate of PTP1C gene expression. The possible role of PTP1C in the macrophage differentiation will be also discussed.

Base Sequence↗

Isolation and characterization of two forms of Xenopus prolactin.

Two forms of highly purified prolactin (PRL) were obtained from pituitary glands of Xenopus laevis by extraction of acetone-dried powder with acid acetone and high-performance liquid chromatography on anion exchange, gel filtration, and reverse-phase columns. Purification was monitored by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis employing antiserum against bullfrog PRL. The Xenopus prolactins (xPRL-I and xPRL-II) thus obtained were shown to have similar molecular weights of 23,000 as determined by SDS-PAGE. The isoelectric points of xPRL-I and xPRL-II determined by isoelectric focusing were 5.6 and 5.3, respectively. Both hormones blocked T4-induced shrinkage of Xenopus tadpole tail fin in vitro. The amino acid compositions of the xPRLs resembled that of bullfrog PRL. The partial amino acid sequences of xPRL-I and of xPRL-II showed 78 and 68% homology with the comparable portion of the sequence of bullfrog PRL, respectively. Homology between xPRL-I and xPRL-II was 90%.

Amino Acid Sequence↗

Immunocytochemical identification of growth hormone (GH) cells in the pituitary of three anuran species using an antiserum against purified bullfrog GH.

An antiserum was prepared against the recently purified bullfrog (bf) growth hormone (GH); it was applied to sections of brain and pituitary of three urodele (Ambystoma, Pleurodeles and Cynops) and three anuran (Xenopus, Bufo vulgaris and B. japonicus) species. No immunostaining was obtained in the urodele pituitary, being consistent with the results of immunoblot analysis of the pituitary homogenate. In the three anuran species, strong immunoreactivity was observed in GH cells that were concentrated in the posterodorsal region of the pars distalis. No GH-like immunoreactivity was detectable in the brain of any of the species. A comparison using adjacent sections stained with anti-bf prolactin (PRL) confirmed the anteroventral localization of PRL cells. Colocalization of GH and PRL was not apparent. These data suggest that the molecular structure of amphibian GHs is considerably different between anurans and urodeles. The antiserum used in the present work shows a high species specificity, recognizing only anuran GHs. In contrast anti-bfPRL labeled PRL cells in all the amphibian species studied in the present work, suggesting that PRLs possess common amino acid sequences recognized by the anti-bfPRL.

Animals↗

Idiopathic chronic calcifying pancreatitis with diabetes mellitus. Analysis of pancreatic stone protein gene.

We present a case of a 27-year-old female suffering from chronic calcifying pancreatitis with diabetes mellitus. Radiographic examinations and exocrine pancreatic function tests revealed considerable dilatation of pancreatic ducts with large intraductal calculi and exocrine pancreatic insufficiency, respectively. Recent literature indicates that a decrease in the activity of pancreatic stone protein (PSP), which inhibits CaCO3 crystal formation in pancreatic juice, is closely related to the development of chronic calcifying pancreatitis. The patient had no apparent cause or family history of pancreatitis. We therefore investigated the possibility that alterations in the PSP gene might explain the chronic pancreatitis seen in this patient. Six exons of the PSP gene amplified by polymerase chain reaction were directly sequenced, but there was no apparent base mutation observed. Furthermore, Southern blot analysis revealed neither rearrangement nor deletion of the PSP gene in the genomic DNA of this case. However, this genetic approach will be useful for future study of the etiology of hereditary pancreatitis.

Adult↗

PGE2 protects isolated cells against injury through multiple mechanisms.

In order to clarify how PGE2 regulates gastric mucosal integrity, we examined the effects of PGE2 on ethanol-caused injury of isolated gastric chief cells, cultured gastric mucous cells from guinea pigs and Balb/c 3T3 fibroblasts. Pretreatment of these cells with PGE2 reduced ethanol-caused injury of the cells. Furthermore, pretreatment of gastric mucous cells with indomethacin enhanced ethanol-caused injury, suggesting that endogenous PGE2 may be involved in the cell protection. PGE2 stimulated an increase in diacylglycerol (DG) accumulation in chief cells and treatment of chief cells with synthetic DG reduced the injury of the cells. However, DG accumulation was not observed in gastric mucous cells treated with PGE2. Therefore PGE2 may protect the cells from injury through a variety of mechanisms. In addition, PGE2 enhanced the survival of the quiescent fibroblasts cultured in the absence of serum, while PGE2 had no survival enhancing effect on gastric mucous cells. These results suggest that the mechanism by which PGE2 preserves the cell viability may depend on not only cell types used but also how the cells are injured.

Animals↗

Re-freeze dried bilayer artificial skin.

A bilayer artificial skin composed of a silicone sheet and a collagen sponge sheet was developed by us in 1986, modifying Yannas and Burke's technique. It was used in experimental animals and clinically, both with success; but the artificial skin was inconvenient for clinical use in terms of disinfection and preservation. In an attempt to reduce the drawbacks, we developed a re-freeze dried artificial skin. Experiments were conducted to examine whether the re-freeze dried material is equivalent to the prototype one. A significant difference between the two was found in tensile tests but not in experimental and clinical effects.

Adult↗

Influence of anesthetic regimens on intestinal absorption in rats.

We compared the influence of anesthetic regimens using urethane (U), pentobarbital (P), ether (E), and ketamine/midazolam (K) on the intestinal absorption of several probes using a single-pass perfusion technique in rats. The selected probes were D-glucose (1 mM) for the resistance of the unstirred water layer (UWL), D-glucose (100 mM) for the capacity of carrier-mediated D-glucose transport, L-glucose, and urea for membrane-limited passive transport, and tritiated water (3H2O) for blood flow at the absorption site. The absorbed fraction of D-glucose (1 mM) was the smallest for U and the largest for P, suggesting that the resistance of UWL is the largest for U and the smallest for P. The absorbed fraction of D-glucose (100 mM) was the largest for P (U = E = K < P), suggesting a higher capacity of carrier-mediated D-glucose transport for P. The absorbed fraction of urea was similar for all anesthetics, while that of L-glucose was the smallest for K (U = P = E > K). Although the results for these two markers of membrane-limited passive transport were inconsistent, the passive permeability of the intestinal membrane may be lower when treating with K. The intestinal absorptions of D-glucose (1 and 100 mM), L-glucose, and urea were, in general, lower with any of the anesthetics than under nonanesthesia (N), suggesting increased resistance of UWL and decreased intestinal membrane permeability by carrier-mediated and passive transport under anesthesia. The only exception was the absorption of D-glucose (100 mM) under P, which was comparable to that under N.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthetics↗

Synergic activity of imipenem/cilastatin combined with cefotiam against methicillin-resistant Staphylococcus aureus.

The synergic activity of imipenem/cilastatin combined with cefotiam was studied in a mouse bacteraemia model. Combinations of imipenem plus cefotiam in ratios from 1:5 to 1:160 were more effective than either imipenem alone or cefotiam alone (P < 0.05). Synergy was observed against both beta-lactamase producing and beta-lactamase non-producing MRSA. Staggered combinations of imipenem with cefotiam (each drug was administered at a different time) were studied in an in-vitro pharmacokinetic system to clarify relationships between killing kinetics and pharmacodynamics of the combinations. In the in-vitro system, cefotiam (1 g over 30 min) administered 2 h after imipenem administration (250 mg over 30 min) reduced viable cell counts to an undetectable level and maintained this for 4 h, while the simultaneous administration of imipenem and cefotiam maintained an undetectable cell count for only 2 h. Furthermore, imipenem administered after cefotiam showed no synergy. These results indicate that the timing of dosing of each antibiotic influences synergy, and administration of cefotiam 2 h after imipenem is more effective than the other regimens.

Animals↗

Localizing value of seizure manifestations of temporal lobe epilepsies and the consequence of analyzing their sequential appearance.

We investigated the localizing and lateralizing value of principal seizure manifestations in temporal lobe epilepsies (signal symptoms, oroalimentary automatisms, somatomotor manifestations, unilateral dystonic posturing, ictal speech, motionless stare) of 223 complex partial seizures in 50 patients. All the patients had invasive long-term monitoring with the combined implantation of intracerebral electrodes in and subdural electrodes on the bilateral temporal lobes. Postoperative freedom from seizures was ascertained for longer than one year. We found that 35 patients had amygdalohippocampal seizures and 15 had lateral temporal seizures. The value of the manifestations was established in relation to the site and side of seizure origin and to the progression of seizure discharges within the unilateral temporal lobe or to the contralateral cerebral hemisphere. Several signs among the manifestations were found to be reliable in predicting the site or side of the temporal lobe seizure focus. We emphasized the importance of investigating sequential changes of seizure manifestations in relation to ictal EEG findings by means of simultaneous recording.

Amygdala↗

The effect of nicardipine on maternal and fetal hemodynamics and uterine blood flow in chronically instrumented pregnant goats.

This study was designed to clarify the maternal and fetal hemodynamics of nicardipine (0.02 mg/kg) administered intravenously to unanesthetized, chronically instrumented pregnant goats. Nicardipine produced a significant increase in maternal heart rate and cardiac output, and a significant decrease in maternal mean arterial blood pressure and systemic vascular resistance. These changes persisted for about 30 minutes. There were no consistent changes in maternal pulmonary hemodynamics. Uterine arterial blood flow (UBF) decreased significantly (about 15%) after 5 minutes of the injection and returned to the control value within 30 minutes. A significant correlation between the decreases in UBF and maternal diastolic blood pressure was observed after 5 minutes of the injection. The fetal heart rate, blood pressure and acid-base status did not change after the injection of nicardipine. These observations of pregnant goats suggest that nicardipine might be a useful agent for the treatment of acutely hypertensive crises in pregnant patients.

Animals↗

In vitro activity of a new carbapenem antibiotic, BO-2727, with potent antipseudomonal activity.

BO-2727, a new 1-beta-methyl-carbapenem, was active at concentrations of 6.25 micrograms/ml or less against gram-positive and gram-negative bacteria, including some imipenem- and/or meropenem-resistant (MICs, > or = 12.5 micrograms/ml) Pseudomonas aeruginosa strains, against which it proved generally fourfold more active than imipenem and meropenem. BO-2727's antipseudomonal activity and its broad spectrum merit further investigation for clinical use by itself, since it was stable in the presence of renal dehydropeptidase I.

Anti-Bacterial Agents↗

Human T lymphotropic virus type I associated myelopathy with pulmonary and cutaneous lesions.

A necropsy case of human T lymphotropic virus I (HTLV-I) associated myelopathy (HAM) in a 64 year old man with serological and genetical confirmation of HTLV-I infection is reported. The spinal cord, lung, and skin were mainly affected. Severe degeneration had occurred in the spinal cord, not only in the lateral columns but also in the anterior and posterior columns. The degenerate lesions showed proliferation of capillaries, loss of myelin and axon, and perivascular and parenchymal infiltration with T lymphocytes and foamy macrophages in the white matter. T lymphocytes had infiltrated the lung and there was vascular proliferation in the peribronchus. OPD4 positive cells predominated in the lung. The patient also had erythrodermia where dense and bandlike HTLV-I infected lymphoid cell infiltration was observed, with mild atypia and epidermotropism. HTLV-I may cause multiorganic inflammatory disorders, although the definitive role of HTLV-I in the pathogenesis is still unknown.

Bronchial Diseases↗

Androgen regulation of thromboxane A2/prostaglandin H2 receptor expression in human erythroleukemia cells.

Thromboxane A2 (TxA2), a platelet aggregator and vasoconstrictor, has been implicated as a potential mediator of cardiovascular diseases. Abuse of androgenic steroids has been associated with thrombotic cardiovascular diseases. Human erythroleukemia (HEL) cells, a megakaryocyte-like cell line, express functional TxA2/prostaglandin H2 (PGH2) receptors with characteristics similar to those seen in platelets. This study characterized testosterone regulation of HEL cell TxA2/PGH2 receptors. TxA2/PGH2 receptor affinity (Kd) and density (Bmax) were determined via equilibrium binding experiments using the radiolabeled TxA2 mimetic (1S-[1 alpha,2 beta(5Z),3 alpha(1E,3R*),4 alpha])-7-(3-[3-hydroxy-4-(4'- iodophenoxy)-1-butenyl]-7-oxabicyclo[2.2.1]heptan-2-yl)-5-he ptenoic acid (125I-labeled BOP). Testosterone (200 nM) but not estradiol increased Bmax from 108 +/- 9 fmol/mg protein to 157 +/- 9 fmol/mg protein (n = 7 experiments; P < 0.01) without any significant change in Kd. Testosterone had no significant effect on alpha 2-adrenergic receptor density. The maximum increase in intracellular free calcium induced by the TxA2 agonists I-BOP or U-46619 was significantly (P < 0.005) greater in testosterone-treated cells compared with controls. Hydroxyflutamide (1 microM), an androgen-receptor antagonist, completely blocked the effect of testosterone (P < 0.01). Dihydrotestosterone, the active metabolite of testosterone, also increased Bmax in a concentration-dependent manner and was more potent than testosterone. The effect of testosterone to increase Bmax was significantly (P < 0.01) inhibited by coincubation with cycloheximide (0.1 microgram/ml) or actinomycin D (10 ng/ml). These results indicate that androgenic steroids regulate the expression of functional TxA2/PGH2 receptors in HEL cells. These findings may have relevance to cardiovascular disease.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Cloning and characterization of K562 cells on hemoglobin synthetic activity.

Two clones of the K562 human leukemic cell line were isolated according to hemoglobin (Hb) expression. One clone was expressed less than 5% (K562-L) and the other more than 90% (K562-H). The two clones did not exhibit any difference in cell growth or cell cycle. However, the Hb expression of K562-H cells was reduced by succinylacetone (S.A.). The above results suggested that the difference in the Hb production of K562-L and K562-H cells depended on the heme synthetic activity. On the other hand, glycophorin A was expressed to a greater extent on K562-L cells than on K562-H cells. These findings suggested that heme synthesis and the expression of glycophorin A on K562 cells were not always related. The CD11b and the CD61 were also expressed to a greater extent on K562-L cells than on K562-H cells, but the CD34 was not expressed on these cells.

Antigens, CD↗

Relationship between molecular state of serum HBV-DNA and clinical features of hepatitis B virus carriers.

Molecular species of serum hepatitis B virus (HBV)-DNA in HBV carriers were classified by Southern blot hybridization into three types; type I with two bands of 4.0 kb and 3.2 kb, type II with the two bands of type I plus the smear between 4.0 kb and 3.2 kb, and type III with a broad band below 4.0 kb. A total of 51 HBV carriers were classified into three groups (group I, n = 19 with type I; group II, n = 12 with type II; and group III, n = 20 with type III). Serum aminotransferase levels of group I were significantly lower than those of groups II and III. Liver pathology revealed that 18 of the 19 (94.7%) group I patients showed nonspecific reactive hepatitis (NSRH), while 11 of the 12 (91.7%) group II patients and 19 of the 20 (95.0%) group III patients showed chronic persistent hepatitis (CPH) or chronic active hepatitis (CAH). Immunohistochemical study showed that hepatitis B core antigen (HBcAg) was localized in the nucleus of hepatocytes in most of patients with type I while it was localized in both the nucleus and cytoplasm in those with types II and III. Since the smear between 4.0 kb and 3.2 kb is specifically found in groups II and III, HBV-DNA with this smear may be related to active hepatitis.

Adult↗