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Biomedical subjects

T Ide

Publications and source records attributed to T Ide.

At least 181 records · Page 10Linked to original sources
PubMed1996

[Trials and analysis of umbilical cord blood collection, separation and cryopreservation methods for transplantation].

Placental and umbilical cord blood, as an alternative course of haematopoietic stem cells for bone marrow reconstitution, have recently been showed to yield successful sibling-donor cord blood grafts children. The advantages of using cord blood are related to the high number of haematopoietic progenitors in circulation at birth. In our study there was a remarkable heterogeneity of volume, number of nucleated cells and the number of progenitors from sample to sample. Seven out of 40 samples of more than 61 ml blood volume contained 1.8 (+/- 1.0) x 10(5) CFU-GM or 6.0 (+/- 4.8) x 10(8) nucleated cells. 10 ml of whole blood was necessary for laboratory tests including ABO blood type, screening of infectious markers, HLA typing, as well as frozen sera and cells for the possible future tests. Collected cord blood of more than 70ml in volume may have sufficient numbers of CFU-GM for engraftment to pediatric patients weighing about 20 kg. A cord blood bank project is now going on by collecting blood with informed consent of the mother in cooperation with obstetrics and gynecology staff.

ABO Blood-Group System↗
PubMed1996

Telomerase in hepatocellular carcinogenesis.

Despite the recent advances in diagnostic techniques of HCC, diagnosis of HCC is still difficult and ambiguous when HCC is small and of the well differentiated type. The results presented here demonstrated that strong telomerase activity was frequently detected in HCC irrespective of the stage or size of the nodules but neither in non-tumor diseased liver nor in normal liver. Telomerase activity determination can be a useful additional tool for the diagnosis of early well-differentiated HCC.

Biomarkers, Tumor↗
PubMed1995

Increase in expression levels of interferon-inducible genes in senescent human diploid fibroblasts and in SV40-transformed human fibroblasts with extended lifespan.

The normal human fibroblast line, TIG-3 which senesces at around 80 population doubling levels (PDLs), expressed interferon (IFN)-inducible genes such as 6-16, 2', 5'-oligoadenylate synthetase (2,5-A) and HLA B7 near the end of the proliferative lifespan. Other normal fibroblast line such as MRC-5 also expressed IFN-inducible genes when senesced. Clones transformed with SV40 T-antigen, which extended their proliferative lifespan by about 20-30 PDLs, also expressed IFN-inducible genes during their extended life. Anti-IFN-beta antibodies added in culture medium repressed the expression of IFN-inducible gene in both normal senescent and life-extended SV40-transformed cells. IFN-beta repressed DNA synthesis in normal TIG-3 and induced IFN-inducible genes in both normal and SV40-transformed TIG-3. Conditioned medium recovered from life-extended SV40-transformed cells contained IFN-beta, but not IFN-alpha, IFN-gamma or TNF-alpha and possessed an activity that inhibited DNA synthesis of young TIG-3. Addition of anti-IFN-beta antibodies into the medium enhanced the serum-induced DNA synthesis of near senescent (91% lifespan completed) TIG-3, while it neither induced DNA synthesis in fully senescent TIG-3 nor extended the proliferative lifespan of TIG-3. These results suggest that normal and SV40-transformed human fibroblasts increase expression of IFN-beta with increasing proliferative age especially near the end of their lifespan resulting in induction of IFN-inducible genes and possibly in growth repression.

Cell Line↗
PubMed1995

Inhibitors of cGMP-dependent protein kinase block senescence induced by inactivation of T antigen in SV40-transformed immortal human fibroblasts.

Immortal human fibroblasts isolated following transfection with thermolabile simian virus 40 T antigen lost division potential upon shift up in temperature due to heat inactivation of the antigen. Such cells showed a concomitant change in the distribution of a mortality marker, mortalin, from a juxtanuclear cap like distribution of immortal cells to a uniform cytosolic distribution of mortal cells. We made an attempt to modulate the above inducible system of cellular senescence using various protein kinase inhibitors. Among the indolocarbazole type inhibitors tested, only KT5823, defined as a specific inhibitor of cGMP-dependent protein kinase, blocked the loss of division potential as determined by cell growth and colony forming ability. This inhibitor also prevented the above change in mortalin distribution due to temperature shift. In addition, the isoquinoline sulfonamide derivatives H8, H9, H88 and H89, all shown to inhibit cGMP-dependent protein kinase, suppressed the senescence. Inhibitors specific to other types of protein kinases, protein phosphatases or tyrosine kinases tested had no effect. Since there was no difference between the effective and non-effective inhibitors in their effects on cell cycle progression, cell cycle arrest by itself cannot account for the above phenomenon. These results suggest that a signaling pathway possibly mediated by cGMP-dependent protein kinase is involved in the induction of cellular senescence.

Alkaloids↗
PubMed1995

An anion channel from transverse tubular membranes incorporated into planar bilayers.

Transverse tubular (TT) vesicles from rabbit skeletal muscle were incorporated into planar lipid bilayers to characterize the chloride channel. The single channel conductance of the channel was 40 pS in choline-Cl solution (cis, 300 mM/100 mM, trans). The gating rate of the channel does not depend on membrane voltage. The channel was blocked by stilbene derivatives (DIDS and SITS), which are known as inhibitors of voltage-dependent Cl- channels of the Torpedo electric organ, from both sides of the membrane. An inhibitor of voltage-dependent Cl- channels of skeletal muscles, 9-anthracene carboxylic acid (9-AC) inhibited the channel from the cis side of the membranes, which corresponded to the cytoplasmic space. Ethacrynic acid (EA), which is reported to inhibit Cl- conductance of the kidney and trachea, decreased the open probability of the TT Cl- channel concentration dependently. Indanyloxyacetic acid (IAA), which is also reported to be an inhibitor of kidney and trachea Cl- channels, decreased the single channel current without affecting open probability of the TT Cl- channel.

Animals↗
PubMed1995

Enhanced expression of cyclin D1 in senescent human fibroblasts.

When human fibroblast, TIG-1, was growth-stimulated with fetal bovine serum, the induction level of cell cycle-dependent genes was generally much lower in senescent cells than in young counterparts. Exceptionally, the expression level of cyclin D1 in senescent cells was constitutively higher than in young cells and further increased after serum stimulation, which was confirmed by Northern and Western blots and immunoprecipitation. This was also true in other human diploid fibroblast lines, TIG-3 and MRC-5. However, cyclin D1-dependent kinase activity was not detected in senescent cells. When sense- or antisense-cyclin D1 cDNA driven by beta-actin promoter was transfected into young TIG-1 cells, the number of appeared colonies from sense-strand transfected cultures was lower than that from antisense-strand-transfected ones. However, clones expressing cyclin D1 at low or undetectable level which were isolated after transfection with antisense-cyclin D1 proliferated up to the same division limit as untransfected and sense-strand transfected cells. Four clones of SV40-transformed TIG-1 expressed cyclin D1 at moderate levels during their extended proliferative lifespan. It appears that, if the extremely overexpressed cyclin D1 could cause an inhibition of cell proliferation at senescent stage, cellular senescence occurs regardless of overexpression of cyclin D1.

Animals↗
PubMed1995

Telomerase activity in human liver tissues: comparison between chronic liver disease and hepatocellular carcinomas.

Telomerase activity was examined in 105 frozen samples from human normal liver tissues, chronic liver disease, and hepatocellular carcinoma (HCC). Telomerase activity was positive in 28 of 33 HCC tissues regardless of tumor stage or size. Telomerase was expressed in 15 of 18 differentiated HCC nodules smaller than 3 cm. HCC tissues from all eight hepatitis B virus-positive patients were telomerase positive, while telomerase activity was not detected in normal liver tissues (0 of 4). Weak telomerase activity was only detected in 1 of 22 nontumor liver tissues from HCC patients. Interestingly, in 19 of 38 hepatitis tissues and 6 of 8 cirrhotic liver tissues from apparently cancer-free patients, very weak telomerase activity was detected. These results indicate that the expression of telomerase may play a crucial role in hepatocarcinogenesis.

Base Sequence↗
PubMed1995

Increase in expression level of p21sdi1/cip1/waf1 with increasing division age in both normal and SV40-transformed human fibroblasts.

p21sdi1/cip1/waf1 gene, whose product inhibits Cyclin/Cdk kinase and blocks cell proliferation, is known to be up-regulated by p53 and to be expressed at high levels in senescent human fibroblasts. The present paper reports that p21 expression level was much lower in SV40tsT-antigen transformed cells than in normal cells and that after inactivation of tsT-antigen it recovered to that of normal cells. However, cell division age-dependent increase in expression level of p21 occurred not only in normal human fibroblasts but also in SV40tsT-antigen transformed fibroblasts with active T-antigen. These results suggest that cell age-dependent increase in p21 occurs independently upon the function of T-antigen binding proteins including p53, while the basal expression level of p21 is up-regulated by p53 as previously reported.

Antigens, Polyomavirus Transforming↗
PubMed1995

Effects of hydroxyapatite and alumina sheaths on postoperative peritendinous adhesions in chickens.

Two experimental methods for restoring flexor tendon sheath integrity and preventing adhesions around traumatized flexor tendons utilizing artificial tendon sheaths made of either hydroxyapatite (HAp) or alumina were studied in a flexor tendon-trauma model and compared to a standard tendon sheath repair and a control. Eighty toes were divided equally into a control group, a sheath repair group, an HAp group, and an alumina group. Profundus tendons in zone II were divided and repaired after sublimis excision in all groups. In the sheath repair group, the flexor sheath was also repaired after suturing the tendon. In artificial sheath groups, sheaths made of HAp and alumina were placed over the repair sites to protect them from the surrounding tissues. In the control group, after repairing the tendon, the flexor sheath was excised and no artificial sheaths were used. Each toe was immobilized in a plaster cast for 3 weeks. After three weeks, the plaster cast was removed followed by the removal of the sheaths in the artificial sheath groups through a small incision in the skin in zone II. Active mobilization was encouraged in each group. Postoperative adhesions were examined at 3, 6, 9, and 12 week intervals by using light microscopic techniques. To further explore the effects of artificial sheaths on tendon healing, transmission electron microscopy was done for the HAp and alumina groups at 3, 6, and 12 week intervals. Results demonstrated decreased severity of postoperative adhesions in the HAp as well as in the alumina groups in comparison with the sheath repair and controls. A space resembling the fibro-osseous canal was formed around the tendon after removing the sheaths. This space remained patent until 12 weeks, 9 weeks after removing the sheaths, and a newly formed tendon sheath-like structure lined by synovial cells and with a peritenon-like structure over the tendon surface was observed. In the sheath repair and control groups, the severity of adhesions was decreased with the passage of time, to some extent due to unrestricted mobility. However, a newly formed tendon sheath or peritenon-like structure was not observed. Electron microscopic studies confirmed good healing at the suture in the HAp and alumina groups with no evidence of necrosis. These results are qualitative in nature as no statistical tests were performed. From these results we conclude that if the tendon is separated from the surrounding granulation tissue by a barrier with good biocompatibility, the tendon can heal with fewer adhesions.(ABSTRACT TRUNCATED AT 400 WORDS)

Aluminum Oxide↗
PubMed1995

Transforming growth factor-beta 1 has both promoting and inhibiting effects on induction of DNA synthesis in human fibroblasts.

Growth-arrested human normal fibroblasts, TIG-1, initiated DNA synthesis following addition of epidermal growth factor (EGF). Transforming growth factor-beta 1 (TGF-beta 1) by itself had no effect on induction of DNA synthesis. When EGF and TGF-beta 1 were added simultaneously to growth-arrested TIG-1 cells, induction of DNA synthesis was enhanced compared with that by EGF alone. Contrarily, when TGF-beta 1 was added earlier than 2 h or later than 2 h of EGF addition, induction of DNA synthesis was prevented. Induction of DNA synthesis by EGF was insensitive to pertussis toxin (PT, an inhibitor of Gi protein) and to staurosporine (a protein kinase inhibitor). The promoting effect of TGF-beta 1 on DNA synthesis was PT-insensitive and staurosporine-insensitive. Contrarily, inhibitory activity of TGF-beta 1 on DNA synthesis was PT-sensitive and staurosporine-insensitive. These studies suggest that the effect of TGF-beta 1 is to promote or to inhibit induction of DNA synthesis by EGF expressed through different signal transduction processes in the same cell.

Alkaloids↗
PubMed1995

Induction of gadd153 gene in G0-specific ts mutant cells at nonpermissive temperature.

tsJT60 is a temperature-sensitive (ts) mutant of the cell cycle isolated from a rat cell line. It does not enter the cell cycle from the G0 phase at a nonpermissive temperature (40 degrees C) but proliferates normally in log phase at both permissive (34 degrees C) and nonpermissive temperatures (G0-specific ts mutant). Here we examined whether tsJT60 fails to downregulate growth arrest-specific genes after growth stimulation from G0 phase at 40 degrees C. Among growth arrest-specific genes examined, an expression of gadd153, which decreased after serum stimulation at 34 degrees C as expected, increased markedly at 40 degrees C. It is plausible to assume that induction of gadd153 at 40 degrees C is correlated with inability of tsJT60 to enter S phase from G0 phase at 40 degrees C, because it also occurred in tsJT366 (another ts mutant clone in the same complementation group) but did not occur in parental 3Y1 and in other ts mutant lines of different complementation groups. However, gadd153 was also induced after the temperature upshift in tsJT60 cells growing in log phase or progressing S phase.

Animals↗
PubMed1995

A cDNA clone encoding an IgE-binding protein from Brassica anther has significant sequence similarity to Ca(2+)-binding proteins.

Thirteen cDNA clones encoding IgE-binding proteins were isolated from expression libraries of anthers of Brassica rapa L. and B. napus L. using serum IgE from a patient who was specifically allergic to Brassica pollen. These clones were divided into two groups, I and II, based on the sequence similarity. All the group I cDNAs predicted the same protein of 79 amino acids, while the group II predicted a protein of 83 amino acids with microheterogeneity. Both of the deduced amino acid sequences contained two regions with sequence similarity to Ca(2+)-binding sites of Ca(2+)-binding proteins such as calmodulin. However flanking sequences were distinct from that of calmodulin or other Ca(2+)-binding proteins. RNA-gel blot analysis showed the genes of group I and II were preferentially expressed in anthers at the later developmental stage and in mature pollen. The recombinant proteins produced in Escherichia coli was recognized in immunoblot analysis by the IgE of a Brassica pollen allergic patient, but not by the Ige of a non-allergic patient. The cDNA clones reported here, therefore, represent pollen allergens of Brassica species.

Amino Acid Sequence↗
PubMed1995

Octadecatrienoic acids as the substrates for the key enzymes in glycerolipid biosynthesis and fatty acid oxidation in rat liver.

The activities of key enzymes in glycerolipid biosynthesis and fatty acid oxidation were compared using CoA esters of naturally occurring positional isomers of octadecatrienoic acids (18:3) as the substrates. The trienoic acids employed were 9,12,15-18:3 (alpha-18:3), 6,9,12-18:3 (gamma-18:3), and 5,9,12-18:3 (pinolenic acid which is a fatty acid contained in pine seed oil, po-18:3). The activities of microsomal glycerol 3-phosphate acyltransferase obtained with various 18:3 were only slightly lower than or comparable with those obtained with palmitic (16:0), oleic (18:1), and linoleic (18:2) acids. Mitochondrial glycerol 3-phosphate acyltransferase was exclusively specific for saturated fatty acyl-CoA. The activities of microsomal diacylglycerol acyltransferase measured with various polyunsaturated fatty acyl-CoAs were significantly lower than those obtained with 16:0- and 18:1-CoAs. Among the polyunsaturated fatty acids, gamma-18:3 gave the distinctly low activity. The Vmax values of the mitochondrial carnitine palmitoyltransferase I were significantly higher with alpha-18:3 and po-18:3 but not gamma-18:3, than with 16:0 and 18:2, while the apparent Km values were the same irrespective of the types of acyl-CoA used except for the distinctly low value obtained with gamma-18:3. The response to an inhibitor of the acyltransferase reaction, malonyl-CoA, was appreciably exaggerated with 18:2, alpha-18:3, and po-18:3 more than with 16:0 and 18:1. However, the response with gamma-18:3 was the same as with 16:0. Thus, some of glycerolipid biosynthesis and fatty acid oxidation enzymes could discriminate not only the differences in the degree of unsaturation of fatty acids but also the positional distribution of double bond among the naturally occurring 18:3 acids.

Acyltransferases↗
PubMed1995

Effect of aminophylline on high-energy phosphate metabolism and fatigue in the diaphragm.

BACKGROUND: Diaphragmatic fatigue causes respiratory failure, for which aminophylline has been used as therapy. Because the mechanism of action of aminophylline in reversing diaphragmatic fatigue is unclear, we used in vivo 31P magnetic resonance spectroscopy (MRS) to determine the relation between diaphragmatic activation, force output, and aerobic metabolism. METHODS: Bilateral phrenic stimulation was used to pace the diaphragm in pentobarbital-anesthetized piglets (6-10 weeks old; n = 44). Esophageal and abdominal pressures were measured to calculate transdiaphragmatic pressure (Pdi) (Pdi = abdominal pressure-esophageal pressure) as an index of force output. Activation was determined by the amplitude of the compound action potential of the diaphragmatic electromyogram. Aerobic metabolism was assessed with a 31P MRS surface coil on the right hemidiaphragm with the animal in a 4.7-T magnet. The animals were divided into four groups based on aminophylline loading dose: saline, aminophylline 10 mg/kg (A10), aminophylline 20 mg/kg (A20), and aminophylline 40 mg/kg (A40). After aminophylline loading the diaphragm was paced for 25 min followed by a 10-min recovery. RESULTS: Aminophylline concentrations were 12.2 +/- 0.7, 21.9 +/- 2.4, and 44.9 +/- 3.6 mg/l in the A10, A20, and A40 groups, respectively. Compound action potential amplitude decreased in all groups by 30% after 25 min of pacing. Conversely, Pdi remained at 100 +/- 3% of the initial value after 5 min of pacing in the A40 group but decreased to 75 +/- 3% in the saline group. Pdi recovered completely (103 +/- 17%) in the A40 group but remained depressed (72 +/- 6%) in the saline group. Pdi values were intermediate in the A10 and A20 groups. MRS data revealed inadequate energy supply/demand ratio in the saline group such that the ratio of inorganic phosphate to phosphocreatine (Pi/PCr) increased to 1.01 +/- 0.09 after 5 min of pacing. Pi/PCr remained unchanged in the A40 group and was intermediate in the A10 and A20 groups. beta-Adenosine triphosphate and intracellular pH did not differ among groups or as a function of pacing. Diaphragmatic blood flow increased from a resting value of 35-60 to 300-410 ml.min-1 x 100 g-1 during pacing in all groups and was not affected by aminophylline dose. CONCLUSIONS: Aminophylline, in a dose-dependent fashion, delays the onset of fatigue and improves recovery from fatigue. Delayed fatigue is associated with improved aerobic metabolism as reflected in a low Pi/PCr ratio.

Action Potentials↗
PubMed1995

Restoration of the cellular senescence program and repression of telomerase by human chromosome 3.

Telomeres, at the end of chromosomes, shorten with each cell division, resulting in cellular senescence. Tumor cells, unlike normal somatic cells, express a telomerase that maintains the telomere length. Deletion of a gene(s) on chromosome 3 is common in human renal cell carcinoma (RCC) and reintroduction of a normal chromosome 3 into an RCC immortal cell line restored the program of cellular senescence. The loss of indefinite growth potential was associated with the loss of telomerase activity and shortening of telomeres in the RCC cells with a normal chromosome 3. However, microcell hybrids that escaped from senescence and microcell hybrids with an introduced chromosome 7 or 11 maintained telomere lengths and telomerase activity similar to those of the parental RCC23. Thus, restoration of the cellular senescence program by chromosome 3 is associated with repression of telomerase function in RCC cells.

Base Sequence↗