Sequential production of CEA and CEA-related antigens in three-dimensional culture of human gastric carcinoma cells.
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Animals↗
Biomedical subjects
M Matsuda
Publications and source records attributed to M Matsuda.
DNA fragmentation and CEA production in collagen gel culture of human colon carcinoma cells.
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Carcinoembryonic Antigen↗
The enterotoxin of Clostridium perfringens type A binds to the presynaptic nerve endings in neuromuscular junctions of mouse phrenic nerve-diaphragm.
The enterotoxin of Clostridium perfringens type A, a channel-pore forming protein toxin, inhibited neuromuscular transmission in isolated mouse phrenic nerve-diaphragm preparation at low concentrations of calcium. We investigated immunohistochemically the localization of the binding sites of the enterotoxin in the preparation under the conditions in which the enterotoxin reduced maximally the amplitudes of the twitch tension elicited by electrical stimulations to the phrenic nerve. Under the conditions, double immunohistochemical staining of the preparation with (1) rabbit anti-enterotoxin IgG-rhodamine-labeled goat anti-rabbit IgG and (2) mouse anti-synaptophysin (one of the synaptic vesicle-specific membrane proteins)-fluorescein isothiocyanate (FITC)-labeled goat anti-mouse IgG showed that the enterotoxin binds specifically to most of the sites which were stained with anti-synaptophysin exactly in the same configurations having the shapes of the nerve endings in the endplates. The thin section electron micrographs of the enterotoxin-intoxicated preparation showed no alterations in the ultrastructure of the neuromuscular junction and the nerve endings filled with numerous synaptic vesicles. The present results, together with our previous electrophysiological findings, indicate that the enterotoxin binds specifically to the presynaptic nerve endings and inhibits neurotransmitter release at the neuromuscular junction.
Animals↗
Coronary vasoconstriction is the most probable cause of death of rats intoxicated with botulinolysin, a hemolysin produced by Clostridium botulinum.
The pathophysiology of lethal intoxication by botulinolysin (Blyn) was studied using anesthetized rats and isolated rat organs. Intravenous injection of 10,000 and 1000 hemolytic units (HU) of Blyn killed rats rapidly while 100 HU of the toxin did not. Congestion and edema of lungs were observed at autopsies of the rats killed by intoxication. Hemoglobinemia was obvious in rats injected with 1000 HU of Blyn but not in rats with 10,000 HU. Electrocardiograms of the intoxicated rats showed depression of T waves but not changes characteristic of hyperpotassemia. All the rats injected with the above doses of Blyn showed a rapid fall in arterial blood pressure (BP) immediately after the toxin injection, and BP soon recovered in rats injected with 100 HU, partially and transiently in rats with 1000 HU, and not in rats with 10,000 HU of Blyn. Perfusion of Blyn (1 HU/ml) to isolated rat hearts caused a rapid and marked increase in perfusion pressure and cessation of spontaneous heart beat. Acetylsalicylic acid (10(-3) M) and quinacrine dihydrochloride (10(-5) M) did not essentially influence the effects of Blyn on the isolated hearts, but verapamil (10(-6) M) inhibited at least the initial increase in perfusion pressure elicited by Blyn. Spontaneous contractions of the isolated atria were little influenced by Blyn (60 HU/ml). Perfusion pressures of isolated kidneys, lungs and livers were also increased by Blyn (1 HU/ml). The results indicate that Blyn caused vasoconstriction but had little direct effect on myocardium. Based on the above findings, we conclude that coronary vasoconstriction elicited by direct action of Blyn causes acute cardiac dysfunction leading to systemic hypotension and death of the intoxicated animals.
Animals↗
In vitro antiviral activities of sulfated polysaccharides from a marine microalga (Cochlodinium polykrikoides) against human immunodeficiency virus and other enveloped viruses.
A marine microalga, Cochlodinium polykrikoides, produces extracellular sulfated polysaccharides. Isolation and purification of the polysaccharides were accomplished by precipitation with ethanol and Cetavlon, followed by DEAE-cellulose column chromatography (polysaccharides A1 and A2). These polysaccharides, which were homogeneous when analysed by both ultracentrifugal and electrophoretic methods, were composed of mannose, galactose, glucose and uronic acid, together with sulfate groups (S = 7-8% w/w). Both A1 and A2 inhibited the cytopathic effect of influenza virus types A and B in MDCK cells, that of respiratory syncytial virus types A and B in HEp-2 cells, that of human immunodeficiency virus type 1 in MT-4 cells; and, except A1 for herpes simplex virus type 1 and A2 for parainfluenza virus type 2 in HMV-2 cells, the cochlodinium polysaccharides showed no antiviral activity against parainfluenza virus types 2 and 3, measles virus, mumps virus or herpes simplex virus type 1 in HMV-2 cells. No cytotoxicity for host cells was observed with these polysaccharides at a concentration of 100 micrograms ml-1. Inhibitory effects on various viruses were achieved at concentrations that were not markedly inhibitory to the blood coagulation process.
Animals↗
Effects of erythropoietin, IL-3, IL-6 and LIF on a murine megakaryoblastic cell line: growth enhancement and expression of receptor mRNAs.
We examined the effects of recombinant human erythropoietin (rhEPO), recombinant murine interleukin 3 (rmIL-3), recombinant human interleukin 6 (rhIL-6), recombinant human interleukin 11 (rhIL-11), recombinant murine leukemia inhibitory factor (rmLIF) and recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) on the growth of murine megakaryocytic cell lines. In serum-free methylcellulose culture supplemented with bovine serum albumin (BSA), the addition of rhEPO (0.1-10 U/ml), rmIL-3 (10-500 U/ml), rhIL-6 (100-10,000 U/ml), rmLIF (100-10,000 U/ml), or rmGM-CSF (10-1000 U/ml) enhanced colony growth in L8057Y5 cells, which had been maintained in protein-free culture, mostly in a dose-dependent fashion; rhIL-11 did not have any stimulatory effect at the tested doses (10-1000 U/ml). In addition, colony growth of L8057 cells, which had been maintained in serum-containing culture, was enhanced, but to a lesser extent, by the addition of these cytokines except rhEPO (the cultures were supplemented with 1% fetal bovine serum. Among the cytokines that showed growth-enhancing effects on L8057 cells, the expression of mRNAs encoding receptors for EPO, IL-6 and IL-3 was examined by northern blot analysis or reverse transcription polymerase chain reaction (RT-PCR). In both cell lines, mRNAs for EPO-R, IL-6R, gp130, IL-3R alpha and beta chains were constitutively expressed. The results suggest that L8057 and L8057Y5 cell lines have characteristics of megakaryoblastic cells in their biological responses to cytokines, as well as in the expression of cytokine receptor mRNAs, and that the growth-enhancing effects of these cytokines on the cell lines may be achieved through specific receptors. Our findings show the value of these cell lines for investigating the mechanisms of growth signal transduction in megakaryopoiesis.
Animals↗
Thrombomodulin in the sera of patients with multiple sclerosis and human lymphotropic virus type-1-associated myelopathy.
Damage to the blood-brain barrier, which mainly consists of cerebral endothelial cells, has been demonstrated in multiple sclerosis (MS) clinically and histochemically. To investigate the endothelial cell damage, we evaluated the presence of soluble thrombomodulin in the sera of patients with MS and human T lymphotropic virus type-1-associated myelopathy (HAM) using an enzyme-linked immunosorbent assay. Serum thrombomodulin levels were significantly increased in patients with acute relapsing MS during an exacerbation and chronic progressive MS as compared with those of controls (P < 0.001, respectively). Patients with HAM also had higher serum levels of thrombomodulin than did controls (P < 0.001). There was significant difference between patients with HAM and seropositive non-HAM carriers (P < 0.01). These results suggest that the detection of serum thrombomodulin could be used as a marker of endothelial cell damage in inflammatory diseases such as MS and HAM.
Adult↗
Increased levels of soluble vascular cell adhesion molecule-1 (VCAM-1) in the cerebrospinal fluid and sera of patients with multiple sclerosis and human T lymphotropic virus type-1-associated myelopathy.
We evaluated the relationship between the soluble form of vascular cell adhesion molecule-1 (sVCAM-1) and disease activity in patients with multiple sclerosis (MS) or with human T lymphotropic virus type 1-associated myelopathy (HAM), and measured levels of sVCAM-1 in their cerebrospinal fluid (CSF) and sera. Serum and CSF levels of sVCAM-1 were significantly increased in patients with acute relapsing MS during an exacerbation (P < 0.01 and P < 0.001), as well as in chronic progressive MS (P < 0.05 and P < 0.001), compared with healthy individuals and patients with other neurological diseases, respectively. Patients with acute relapsing MS during an exacerbation also exhibited significantly higher serum and CSF levels of sVCAM-1 vs. patients with acute relapsing MS in remission (P < 0.001). Significantly higher serum levels of sVCAM-1 were observed in patients with HAM vs. either healthy individuals (P < 0.01) or non-HAM carriers (P < 0.01). These results suggest that the determination of sVCAM-1 in the sera and CSF may be useful in monitoring the activity of MS and HAM.
Adult↗
Long-term developmental expression and inductivity of cytochrome P450s within intrasplenically transplanted fetal hepatocytes from spontaneously hypertensive rats.
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Animals↗
Production of monoclonal antibodies specific to the carboxyl terminal region of the 85 kDa subunit of phosphatidylinositol 3-kinase: use of the antibodies in recognition of mutant p85.
We have established two hybridomas producing mAb to the carboxyl terminal region of phosphatidylinositol-3 kinase 85 kDa subunit type alpha (p85 alpha). Analysis using deletion mutants of p85 revealed that epitopes for the two mAb were located on the border of the src homology 2 (SH2) sequence located at the carboxyl end of p85. They immunoprecipitated free p85 efficiently, but reactivity to p85 bound to p110 was very weak. Together with the mAb which we have reported previously, a panel of mAb that covered the various parts of p85 alpha was obtained. Using this panel, we characterized two mutants of p85 (70 and 50 kDa) expressed in the human colon carcinoma cell line, HCC2998. No wild-type p85 was detected in these cells. A mAb specific to the carboxyl terminal region detected p70 but not p50, suggesting that this region is missing in p50. The panel of mAb is a useful tool to use to analyse mutant forms of p85.
Animals↗
An ultrasonically powered instrument for laparoscopic surgery: a brief technical report of preliminary success.
We report the use of an ultrasonically powered instrument (CUSA) for laparoscopic surgery. A total of 105 patients underwent laparoscopic or laparoscopic assisted surgical procedures. Ninety-one laparoscopic cholecystectomies (LC), 9 laparoscopic appendectomies (LA), 3 laparoscopic colon resections (LCR), and 2 laparoscopic partial gastrectomies (LPG) were done using CUSA. In LC, CUSA separates the areolar connective tissue between gallbladder and liver bed without dividing any sizable vessels or injuring the liver. In LA, LCR, and LPG, CUSA makes mesenteric vessel identification and division rapid and safe. We concluded that CUSA is useful for laparoscopic surgery.
Adult↗
A novel mutation causing an aberrant splicing in the protein 4.2 gene associated with hereditary spherocytosis (protein 4.2Notame).
We investigated a Japanese patient with protein 4.2 deficiency. SDS-PAGE showed a complete deficiency of protein 4.2, while Western blot analysis revealed a marked decrease in the amount of protein 4.2, and the existence of a doublet of 74 and 72 kDa bands. Direct sequencing and dot-blot hybridization with allele-specific oligonucleotide probes indicated that the proband was compound heterozygous for a missense mutation in codon 142 with Ala-->Thr (GCT-->ACT) and a single nucleotide substitution (G-->A) of the first base of intron 6 (G-->A) of the protein 4.2 gene. The former is the commonest mutation observed in cases of protein 4.2 deficiency, whereas the latter is a novel mutation, located within the consensus sequence of the 5' splicing site (AGGU) (Protein 4.2Notame). RT-PCR analysis using total RNA isolated from reticulocytes of the proband revealed that the intron 6 donor site mutation causes an abnormal splicing; exon 6 is spliced out with intron 6. The abnormal mRNA has a premature termination codon, as the result of a frameshift, and this instability may lead to degradation. Thus, there is a close relation between this mutation and the molecular pathogenesis of protein 4.2 deficiency.
Base Sequence↗
Activation of prothrombin by factor Xa bound to the membrane surface of human umbilical vein endothelial cells: its catalytic efficiency is similar to that of prothrombinase complex on platelets.
Upon incubation of human prothrombin with factor Xa bound to human umbilical vein endothelial cells (HUVEC) (0.5-0.6 fmol factor Xa/10(5) cells), three bonds at Arg273-Thr274, Arg286-Thr287, and Arg322-Ile323 were cleaved, yielding and releasing fragment 1-2 and a degraded form of alpha-thrombin, but not meizothrombin, into the fluid phase. The apparent Km for prothrombin and the Vmax were 0.25 +/- 0.07 microM and 210 +/- 40 fmol thrombin/min/10(5) cells, respectively. For the maximally bound factor Xa, the calculated catalytic efficiency (kcat = 6-7 s-1) was similar to those reported for the prothrombinase complex formed on the phospholipid vesicles and natural membrane surfaces. The prothrombin derivatives lacking the 10 gamma-carboxyglutamic acid (Gla) residues-containing region were not activated by the cell-bound factor Xa. The activation rate of prothrombins with Gla residues variously modified to gamma-methyleneglutamic acids was reduced in accordance with the number of modified residues. For the inhibition of prothrombin activation, intact fragment 1 was needed; the Gla-domain alone did not affect the reaction. Binding of monoclonal antibodies to the region of 1-48 or the kringle 1 region of prothrombin also interfered with the prothrombin activation. Prothrombin activation on the surface of HUVEC appeared to proceed via formation of a cellular prothrombinase complex composed of phospholipids of HUVEC membrane, endogenous factor Va, factor Xa, and prothrombin. The Gla-domain and kringle 1 regions are indispensable for the molecule to serve as an effective substrate for the cell-bound factor Xa.
Amino Acid Sequence↗
Laparoscopic omental patch repair for perforated peptic ulcer.
OBJECTIVE: The authors' initial experience with laparoscopic omental patch repair for perforated peptic ulcer is documented. Its results are compared with those of other procedures and follow-up study is reviewed. SUMMARY BACKGROUND DATA: Since the advent of H2-antagonists, the usefulness of simple closure of a perforated peptic ulcer is increasing, and improvements in laparoscopic surgery have made possible minimally invasive surgery for perforated ulcer. METHODS: From December 1992 to February 1994, laparoscopic omental patch repair followed by use of H2-antagonists was performed successfully in 11 patients. Fifty-five patients underwent other surgical procedures for perforated peptic ulcers (conventional open omental patch: 4, selective vagotomy in combination with antrectomy: 24, distal gastrectomy: 27). RESULTS: The average operation time was 135 minutes. Administration of postoperative pain medication was reduced remarkably (0.9 times per patient), and all patients recovered rapidly. No serious postoperative complications were recorded. After a mean period of 11 months, the postoperative evaluation was satisfactory for all patients, and no ulcer recurrence was found. CONCLUSIONS: In perforated peptic ulcer disease, laparoscopic omental patch repair offers a number of advantages. Because no upper abdominal incision is made, there is decreased postoperative pain, and the patient rapidly recovers with fewer and less severe complications. Although the procedure requires a surgeon with particular expertise in endoscopic suturing technique, surgeons familiar with laparoscopic cholecystectomy can readily perform it after some practice. The authors' preliminary experience suggests that this is a minimally invasive procedure for perforated peptic ulcer that offers an attractive alternative to open surgery.
Adult↗
A rapid latex immunoassay for the detection of plasmin-alpha 2-plasmin inhibitor complex. Utilization of two monoclonal antibodies differentially recognizing respective components of the complex.
Among six monoclonal antibodies raised against the human plasmin-alpha 2-plasmin inhibitor complex (PPI), three antibodies were found to recognize the plasmin part (group 1) and another three the alpha 2-plasmin inhibitor (alpha 2-PI) part (group 2) of the complex. One of the group-1 monoclonal antibodies, designated JIPPI-3, specifically reacted with a segment of plasmin containing kringles 2 and 3. Although all three group 2 antibodies reacted with both alpha 2-PI and PPI on immunoblotting and ELISA, one of them, JIPPI-50, was unable to react with alpha 2-PI, when the antibody had been covalently conjugated to Sepharose 4B gels and tested for reactivity against the antigens in solution. The results indicated that the epitope for this antibody had been buried in nascent alpha 2-PI, but had been exposed by complex formation with plasmin or by possible conformational changes induced in the alpha 2-PI molecule on insolubilization to nitrocellulose membranes or immunoplates. By utilizing a set of JIPPI-3 and JIPPI-50, individually coated onto latex beads, PPI could be measured in plasma in the range of 0.8-100 micrograms/ml without interference by coexisting plasminogen (120-200 micrograms/ml) or alpha 2-PI (70 micrograms/ml). This measurable range seems to cover the level of PPI clinically observed under hyperfibrinolytic states.
Animals↗
Level structure of 114Te.
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Journal Article↗
Effects of Bordetella bronchiseptica dermonecrotizing toxin on bone formation in calvaria of neonatal rats.
The effects of Bordetella bronchiseptica dermonecrotizing toxin on bone formation were investigated using a purified toxin preparation. Single injection of 4.3 ng of the toxin into the subcutaneous tissue overlying the calvariae of neonatal rats necrotized periosteum of parietal bone and degenerated osteoblasts within two days. Nine days after the injection, the lesion of the bone tissue became severe; the bone matrix became thin and fragmented. These observations indicate that dermonecrotizing toxin without other factors produced by the organisms impairs bone formation.
Animals↗
Topical aldose reductase inhibitor for correcting corneal endothelial changes in diabetic patients.
BACKGROUND: Marked variations in cell size (polymegethism) and shape (pleomorphism) are characteristic of the corneal endothelium in diabetic patients and animals. METHOD: Wide field specular microscopy was used to evaluate the clinical efficacy of treating the diabetic corneal endothelium with topical instillation of 0.5% aldose reductase inhibitor, CT-112. RESULTS: Morphological variations (polymegethism and pleomorphism) of the endothelium in eight eyes from eight patients receiving CT-112 resolved within 3 months after initiation of treatment. In contrast, no change in endothelial morphology was noted in five eyes from five patients who received placebo. CONCLUSION: These observations suggest that aldose reductase may be involved in the aetiology of corneal endothelial variations in diabetic patients.
Administration, Topical↗