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Biomedical subjects

E Yoshida

Publications and source records attributed to E Yoshida.

At least 91 records · Page 5Linked to original sources

The utility of an anti-fos ribozyme in reversing cisplatin resistance in human carcinomas.

The results presented here demonstrate that expression of a fos ribozyme limits Fos protein synthesis and enhances sensitivity of A2780DDP cells to antineoplastic agents, including cisplatin. Moreover, the reversal of this resistance is associated with down-regulation of dTMP synthase, DNA polymerase beta, topoisomerase I and hMTII-A, genes previously linked to DNA synthesis and repair. Thus these studies further implicate the role of the c-fos gene in DNA synthesis through modulation of expression of dTMP syntase, DNA polymerase beta and topoisomerase I. Finally, the use of ribozymes to circumvent drug resistance suggests their potential utility as agents to inhibit tumor cell growth.

Antineoplastic Agents↗

Reversal of the malignant phenotype by an anti-ras ribozyme.

In this study a ribozyme (catalytic RNA) was designed to site specifically cleave the mRNA of the activated H-ras gene expressed in human bladder carcinoma EJ cells. The optimal conditions for catalytic cleavage by the ribozyme were demonstrated in vitro. A synthetic DNA encoding the ribozyme was cloned into a mammalian expression vector (pH beta APr-1) and transfected into EJ cells. The expressed ribozyme significantly altered the morphology and suppressed the growth of EJ cells in vitro. These cell lines were examined for their malignant potential in athymic (nude) mice by an orthotopic (transurethral) implantation model, which recapitulates the invasive potential of various bladder carcinomas. EJ tumors expressing the H-ras ribozyme were characterized by a marked reduction in tumor take and invasion compared to those formed by control EJ cells. These differences resulted in almost a twofold increase in survival of mice implanted with ribozyme-containing EJ cells. These results further elucidate the role of ras genes in tumorigenicity and invasion, as well as introduce ribozymes as a new class of anticancer agents.

Animals↗

The induction of cataracts by HIV-1 in transgenic mice.

OBJECTIVE: To elucidate the tissue specificity of the expression of HIV-1 genes in an animal and its pathological effects on these tissues. DESIGN AND METHODS: Transgenic mice carrying a defective HIV-1 genome were bred in order to overcome the host-range barrier of this virus. RESULTS: mRNA specific to the transgene was detected in the eyes and the spleen, and, in smaller quantities, in the thymus and the brain. Interestingly, many of the transgenic mice developed cataracts at 3-6 months of age. Swelling and vacuolation of the lens fiber cells were marked, but the epithelial cells of the lens were less affected. HIV antigens were detected in the lens fiber cells and the retina by immunological staining. Accumulation of large amounts of p24 Gag antigen was demonstrated in the affected lens by immunoblot analysis, while negligible Env or other viral proteins was detected. Although accumulation of the Gag protein was also detected in the skin and the brain, no apparent abnormality was observed in these tissues. CONCLUSIONS: Preferential expression of the HIV genes in the eyes, skin, brain and lymphoid tissues was demonstrated. The accumulation of the Gag protein is suggested to have detrimental effects on lens fiber cells, causing cataracts.

Acquired Immunodeficiency Syndrome↗

Fragmentation hemolysis in a patient with hypertrophic obstructive cardiomyopathy and mitral valve prolapse.

We encountered a 65-year-old female with hypertrophic obstructive cardiomyopathy and mitral valve prolapse who had infective endocarditis and hemolytic anemia. The infecting organism of endocarditis was group A streptococci. With regard to the etiology of the hemolytic anemia, fragmentation hemolysis was considered because fragmented red cells and elevated lactic dehydrogenase were observed. Haptoglobin was markedly decreased. Coombs' test, Ham's test and abnormal hemoglobin were negative. She had not had a hemolytic attack in the past. Ultrasonic cardiography showed asymmetrical septal hypertrophy, mitral valve prolapse and 285 mmHg of calculated pressure gradient in the left ventricle. Cardiac catheterization showed 115 mmHg of left intraventricular pressure gradient and mitral regurgitation (grade 2). Hemolysis was slightly improved after treatment with propranolol. Thus, fragmentation of the normal red cells seemed to be due to shear stress.

Aged↗

[Three cases of young adults advanced pulmonary tuberculosis due to patient's and doctor's delay].

Three young adults advanced pulmonary tuberculosis due to delay of therapy were reported. In case 1 and 3, their delays resulted from difficulty in diagnosis, in case 2, from neglecting medical counseling. Each cases revealed bilateral diffuse shadows on chest roentgenograms on admission, which were typical shadows of advanced pulmonary tuberculosis. Malnutrition might contribute to the development of the diseases, which were improved by anti-tuberculosis therapy and hyperalimentation therapy. These cases were suggested some clinical problems characteristic of pulmonary tuberculosis of young adults.

Adult↗

[A case of severe juvenile pulmonary tuberculosis associated with malnutrition in special reference to nutritional assessment].

We reported a case who had suffered from severe pulmonary tuberculosis in association with severe malnutrition. A 19-year-old man was admitted complaining of high fever, productive cough and body weight loss. Before admission, he worked as a cook for long hours a day and had meals irregularly. On admission, he was cachexic. Coarse crackles were auscultated on the both lung field, and the liver was enlarged. Chest X-ray revealed multiple ringed and mottled shadows mainly in bilateral upper and middle lung field. Laboratory examination revealed inflammatory reaction defined by an increase of white blood cells, accelerated erythrocyte sedimentation rate and positive CRP, impairment of liver function and cell-mediated immunity, hypoxemia and restrictive lung dysfunction. Ziehl-Neelsen stain of sputum showed a large number of acid fast bacilli. The assessment of his nutritional status showed decrease in anthropometric measurements, visceral proteins and the Fischer ratio, which suggested that he was in the status of protein-calorie malnutrition in association with amino acid imbalance. He was treated with nutritional therapy in addition to antituberculous drugs, which improved his clinical symptoms, laboratory data and nutritional assessment. These results suggested the importance of nutritional assessment and nutritional therapy based on it.

Adult↗

Induction of inflammatory arthropathy resembling rheumatoid arthritis in mice transgenic for HTLV-I.

Human T cell leukemia virus type-I (HTLV-I) is the etiologic agent of adult T cell leukemia and has also been suggested to be involved in other diseases such as chronic arthritis or myelopathy. To elucidate pathological roles of the virus in disease, transgenic mice were produced that carry the HTLV-I genome. At 2 to 3 months of age, many of the mice developed chronic arthritis resembling rheumatoid arthritis. Synovial and periarticular inflammation with articular erosion caused by invasion of granulation tissues were marked. These observations suggest a possibility that HTLV-I is one of the etiologic agents of chronic arthritis in humans.

Animals↗

Demonstration of an active component of inter-alpha-trypsin inhibitor in the brains of Alzheimer type dementia.

The putative precursor of A4 amyloid protein associated with Alzheimer's disease is known to have a domain with an amino acid sequence characteristic of a Kunitz-type serine protease inhibitor. Human serum inter-alpha-trypsin inhibitor (ITI) is the most similar inhibitor. We screened brain tissues with senile dementia of the Alzheimer type in an attempt to detect ITI immunoreactivity employing immunohistochemical methods. For this purpose, we used the antibody raised against acid-stable proteinase inhibitor (ASPI) which is an active component of ITI. ASPI immunoreactivity was found to be localized in diffuse type senile plaques, the perivascular area and subpial layer. Reactive astrocytes with intense ASPI immunoreactivity were present in the pyramidal layer of the parahippocampus, where loss of neurons was observed. These findings suggest that ITI may be related to the pathogenesis of Alzheimer type dementia.

Aged↗

Acid-stable protease inhibitor in chronic phase of carrageenin-induced inflammation in rats.

The activity and kinetics of acid-stable protease inhibitor (ASPI) were investigated in the chronic phase of carrageenin-induced inflammation in rats. The ASPI activity was 19.6 +/- 3.1 units/ml in the plasma and 15.4 +/- 2.1 units/ml in the inflammatory exudate. The plasma value was significantly higher than that of the control (11.6 +/- 1.3 units/ml). A kinetics study was performed using purified and radiolabeled rat plasma ASPI, whose NH2-terminal amino acid sequence was Ala-Val-Leu-Pro-Gln-Glu-Asn-Glu-Gly-X-Gly-Ser-Glu-Pro-Leu-Ile-Thr-Gly-Th r-Leu- Lys-Lys-Glu-Asp-Ser-Asn-Gln-Leu-Lys-Tyr-Ser-Glu-Gly-Pro. The half-life of the distributive phase was 4.3 +/- 0.4 min and that of the postdistributive phase (biological half-life) was 42.2 +/- 9.2 min in inflammation. There was no significant difference compared with the values in the control (3.9 +/- 0.4 min and 40.7 +/- 6.5 min, respectively). It appeared that the increase in ASPI in inflammation was not due to prolonged excretion of the inhibitor but to an increased production of it, and ASPI was rapidly distributed to the fluids and tissues.

Amino Acid Sequence↗

Distribution and localization of inter-alpha-trypsin inhibitor and its active component acid-stable proteinase inhibitor: comparative immunohistochemical study.

Inter-alpha-trypsin inhibitor (ITI) is a complex that consists of three components. One of these is the acid-stable proteinase inhibitor (ASPI), which is an acute-phase reactant and a broad-spectrum inhibitor. The tissue distribution of ITI and ASPI were investigated and compared using immunohistochemical methods. ITI immunoreactivity was revealed only in the liver and plasma, while ASPI immunoreactivity was found to be distributed in the brain, liver, kidney, gastrointestinal tract, plasma, and urine. Both immunoreactivities were demonstrated in Kupffer cells of the liver, which is thought to be an ITI-producing organ. From these results, it seems unlikely that ASPI is distributed as a part of the ITI molecule. The residual component of ITI may act as a carrier protein of ASPI, or ASPI in the tissues may be produced independently of ITI.

Alpha-Globulins↗

Activated fibrinolytic enzymes in the synovial fluid during acute arthritis induced by urate crystal injection in dogs.

To identify the relationship of the severity of inflammation and fibrinolytic activity in arthritis, the fibrinolytic activity of synovial fluid was studied in acute experimental arthritis induced by injecting monosodium urate crystals into dogs' knee joints. The maximum activity in the synovial fluid was observed 6 h after crystal injection. It was inferred that the fibrinolytic activity was mainly due to plasminogen activator based on fibrin plate assays, substrate specificity, inhibitor effects and zymography. On the other hand, the activity of lysosomal enzymes (beta-glucuronidase and cathepsin G) reached a peak in the synovia after 12 h. Histological examination of the synovial membrane after 12 h also showed greater inflammation than at 6 h. The peak in fibrinolytic activity preceded the peak of lysosomal enzymes and histological changes. These results suggest that an increase in fibrinolytic activity by plasminogen activator may contribute to the development of an acute inflammatory response.

Acute Disease↗

Characterization of two bacteriocins produced by Clostridium perfringens.

Two types of bacteriocins were shown to be produced in succession by a strain of Clostridium perfringens SN-17. They were separated by diethylaminoethyl cellulose (DEAE) column chromatography at pH 8.5 with a linear concentration gradient of NaCl. One type of bacteriocin (named SN-a) was eluted at 0.07 M and the other type (named SN-b) was at 0.12 M. Each of these was partially purified in a series of column chromatographies: DEAE, Sephadex G-200 (or Bio Gel P-150), and hydroxyapatite. Specific activities of SN-a and SN-b after the last chromatography were at most 30- to 50-fold that of culture filtrate of the organisms. Chromatographed SN-a migrated as a single zone in polyacrylamide gel electrophoresis (PAGE) and the zone showed high biological activity. On the other hand, PAGE pattern of SN-b revealed the presence of a few contamination materials. The activity of SN-b after the last chromatography was hardly recovered from the gel but inactivated SN-b was identified in the gel by examining bacteriocin activity of the DEAE fractions recovered from the gel. The molecular weight of the SN-a and SN-b was determined to be about 70,000 and 100,000, respectively, by molecular sieve chromatography. These bacteriocins were very sensitive to protease but insensitive to DNase and RNase. Bacteriocins were both completely inactivated at 55 C and they were more stable in alkaline pH than in acidic pH. SN-a and SN-b were adsorbed in different ways on the surface of the producer and insensitive strains. Several differences and similarities between these 2 bacteriocins are discussed with special reference to the relationship between them.

Bacteriocins↗

Conversion of biological and immunological properties during a process of decapsulation in a strain of Staphylococcus hyicus.

Repeated subculture at 42 degrees C of Staphylococcus hyicus strain ST67P, which exhibits streaming-type growth in a soft-agar medium, yielded three variants, ST67L, ST67S and ST67C, which had different colonial morphologies; small compact colonies possessing long and short tails and perfect compact colonies. The parent strain and ST67L respectively gave strong and weak positive intensity when stained by rabbit antisera prepared by capsular type I and II strains of Staph. epidermidis conjugated with fluorescein isothiocyanate. Variant ST67L gave a positive result with antiserum prepared by capsular type I strain and no staining was observed with variants ST67S and ST67C against these antisera preparations. Strain ST67C had the lowest virulence although no remarkable difference was shown between the parent strain and variants ST67L and ST67S. The cell volume index of the parent strain was 1.35, 2.43 and 3.71 times larger than those of ST67L, ST67S and ST67C, respectively. The converting activity of rabbit anti-ST67P strain serum absorbed by strain ST67C required four times more of the organisms than strain ST67P, changing the colonial morphology of the strain from diffuse to compact type by the addition of antiserum to soft agar medium. Positive coagulase and false positive clumping factor reaction were shown in variants ST67C, but no remarkable alteration was observed with 19 biochemical properties determined by a conventional identification kit. In ulta-thin sections of the parent strain labelled with rabbit anti-strain ST67P serum conjugated with ferritin, large capsule surrounded by ferritin granules were demonstrated by electron microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Epidermoid cyst derived from an accessory spleen in the pancreas. A case report with literature survey.

A rare case of splenic epidermoid cyst (SEC) of the pancreas discovered in a 32-year-old Japanese female is reported. The lesion, 5 x 6 cm in size including caseous material and serous fluid in the lumen, was discovered by ultrasonography and computed tomography at the tail of the pancreas and was easily removed. Histopathologically, the cystic wall consisted of three components: the inside was lined by mature squamous epithelium with keratinization, the middle layer consisted of splenic pulp with a sinus structure, and the peripheral layer was dense fibrous connective tissue in which some involutional pancreatic ducts and islets were recognized. The literature about SEC of the pancreas is discussed in comparison with other types of epidermoid cyst including lymphoepithelial cyst and dermoid cyst in the pancreas.

Adult↗

Rapid accumulation of plasma acid-stable trypsin inhibitor in experimental acute renal injury.

Acid-stable trypsin inhibitor (ASTI) activity was measured during experimental acute renal tubular dysfunction and glomerulonephritis in rats. A marked elevation of ASTI activity occurred at a very early stage of acute renal tubular damage, and the changes were observed prior to histological abnormalities or elevation of blood creatinine. No alteration in ASTI activity was observed at an early stage of experimental glomerulonephritis. The data obtained confirm that ASTI is excreted through the renal tubules and that the plasma ASTI concentration is very sensitive to renal tubular dysfunction.

Animals↗

Induction of interferon-alpha by interferon-beta, but not of interferon-beta by interferon-alpha, in the mouse.

The antigenicity of the interferon (IFN) produced in transgenic mice carrying an extra mouse IFN-beta gene under the control of mouse metallothionein-I enhancer-promoter was examined after induction with Cd2+. Unexpectedly, IFN-alpha in addition to IFN-beta was detected in the serum. Induction of IFN-alpha was also observed when recombinant mouse IFN-beta was injected into normal mice. IFN-alpha was first detected in the circulation 6-10 hr after the administration of IFN-beta, and after 12 hr, IFN-alpha became the major component of serum IFN. On the other hand, when IFN-alpha was injected, no production of IFN-beta was observed. Messenger RNAs specific for IFN-alpha and endogenous IFN-beta were detected in the spleen, though the amount of IFN-beta mRNA was much less than that of IFN-alpha mRNA. These mRNAs were not detected in other organs including the liver where exogenous IFN-beta gene was markedly expressed. These observations showed that the expression of IFN-alpha is inducible by IFN-beta in the mouse, and the spleen was suggested to be the main site of production. Possible mechanisms of the induction are discussed.

Animals↗

Synthesis of mu-conotoxin GIIIA: a chemical probe for sodium channels.

mu-Conotoxin GIIIA, a 22 amino acid peptide paralytic toxin which inhibits the muscle voltage-activated sodium channels, was synthesized by a solid phase method. No purification of intermediates was necessary for the synthesis, and a simple air oxidation of the deprotected crude peptide gave the desired toxin. By all criteria applied, the synthetic material was indistinguishable from the authentic natural toxin.

Amino Acid Sequence↗