Biomedical subjects

T Deguchi

Publications and source records attributed to T Deguchi.

At least 253 records · Page 14Linked to original sources

PubMed1990

[The expression of ras p21 product in hepatocellular carcinoma].

To elucidate the cell biological significance of ras oncogene, the expression of ras-p21 was analyzed in 53 cases of liver tissues including 34 cases of hepatocellular carcinoma (HCC), by using immunohistochemical method. In result, 22 (65%) cases of 34 HCC and 34 (79%) cases of 43 liver cirrhosis were positive for p21, whereas all of chronic hepatitis and normal livers were negative. Especially, comparative study between the expression of p21 and clinicopathological background of HCC revealed that p21 was prominently expressed in well differentiated form, nodular type, small liver cancer, and the cases showing AFP levels below 400 ng/ml. From these results, it was indicated that ras oncogene might play an important role in malignant transformation of hepatocytes or differentiation of HCC.

Carcinoma, Hepatocellular↗

PubMed1990

Immunogenicity and safety of recombinant yeast-derived hepatitis B vaccine in haemodialysis patients.

The immune response of 32 haemodialysis patients vaccinated with a recombinant yeast-derived hepatitis B vaccine (YHB vaccine) was compared with that of healthy adults and of another haemodialysis patient group vaccinated with the plasma-derived vaccine (PHB vaccine). Twenty-two patients were immunized 3 times (months 0, 1 and 6) intramuscularly with a 20 micrograms dose, and ten patients with a 40 micrograms dose of the YHB vaccine. All members of the former group (100%) and nine of the latter group (90%) were anti-HBs positive after 3 vaccinations (months 7); anti-HBs concentrations with geometric mean titres (GMTs) of 82.3 mIU/ml and 242.1 mIU/ml, respectively, the difference being statistically in significant. The 87 healthy control group participants were vaccinated according to the same schedule but with 10 micrograms per dose, and 98% of them were anti-HBs positive by month 7, with a GMT of 198.3 mIU/ml. The patients responses were higher at each time point than those of 53 dialysis patients vaccinated in an earlier study with either 20 micrograms or 40 micrograms PHB vaccine. Two additional inoculations led to a substantial elevation of the anti-HBs titre in most of the haemodialysis patients; GMTs by month 12 being 506.8 mIU/ml and 979.5 mIU/ml for the 20 micrograms and the 40 micrograms dose respectively. No serious side-effects were observed over the one-year period of study. From these results, it was concluded that the YHB vaccine is highly immunogenic and could replace the PHB vaccine even in dialysis patients.

Adult↗

PubMed1990

Effects of L-2-oxothiazolidine-4-carboxylate, a cysteine pro-drug, on teratogenicity of 5-fluorouracil in mice.

Embryotoxicity and teratogenicity of 5-fluorouracil (5-FU) and modulation of its effect by L-2-oxothiazolidine-4-carboxylate (OTC), a cysteine pro-drug, were evaluated in mice. Pregnant ICR mice were intraperitoneally (i.p.) injected with 25 mg/kg of 5-FU on day 11 of gestation (vaginal plug = day 0). Mice were pretreated i.p. with 950 mg/kg of OTC 4 hours before dosing with 5-FU. Dams were killed on day 17 of gestation. Fetuses were examined for external malformations, especially limb malformations. Pretreatment with OTC decreased the frequency and severity of oligodactyly induced by 5-FU, although the differences were not significant statistically. There was little difference in either liver glutathione levels, or body weight gain during gestation of dams between the 5-FU group and the 5-FU plus OTC group. Fetal mortality and fetal weight of the group treated with 5-FU alone were comparable with those of the group pretreated with OTC. In the present study, teratogenicity of 5-FU seemed to be slightly mitigated with OTC pretreatment.

Animals↗

PubMed1989

Expression and characterization of calmodulin-dependent protein kinase II from cloned cDNAs in Chinese hamster ovary cells.

cDNAs containing the entire coding regions of the alpha and beta subunits of calmodulin-dependent protein kinase II (CaM kinase II) were isolated from a rat cerebrum cDNA library, ligated into an expression vector under the control of SV40 early promoter and introduced into Chinese hamster ovary (CHO) cells. To investigate the role of the alpha and beta subunits and their functional domains in CaM kinase II activity, the properties of the kinases expressed in the transfected cells were studied. CaM kinase II activity was detected in the transfected cells when the alpha and beta cDNAs were introduced into CHO cells simultaneously. RNA transfer blot and protein immunoblot analyses demonstrated the expression of the mRNAs and proteins of both alpha and beta subunits in the cloned cells. When alpha or beta cDNA was introduced into CHO cells separately, a significant level of the enzyme activity was also expressed, indicating that the alpha and beta subunits exhibited enzyme activity individually. The apparent Km values for ATP and MAP 2 were almost the same for the alpha subunit, beta subunit, alpha beta complex, and brain CaM kinase II. However, there was a slight difference in the affinity for calmodulin between the expressed proteins. The alpha and beta subunits expressed in the same cells polymerized to form alpha beta complex of a size similar to that of brain CaM kinase II. The alpha subunit also polymerized to form an oligomer, which showed almost the same S value as that of alpha beta complex and brain CaM kinase II. In contrast, the beta subunit did not polymerize. The alpha subunit, beta subunit, alpha beta complex, and brain CaM kinase II were autophosphorylated with [gamma-32P]ATP in the presence of Ca2+ and calmodulin, which resulted in the appearance of Ca2+-independent activity. The Ca2+-independent activity was 60-75% of the total activity as measured in the presence of Ca2+ plus calmodulin. To examine the functional relationship of peptide domains of the subunits of CaM kinase II, deleted cDNAs were introduced into CHO cells and the properties of the expressed proteins were studied. In cells transfected with alpha or beta cDNA from which the association domain was deleted, a significant level of kinase activity was expressed. However, the expressed proteins showed hardly any autophosphorylation and the appearance of Ca2+-independent enzyme activity was very low, indicating that the association domain was essential for the autophosphorylation and for the appearance of the Ca2+-independent activity.

Animals↗

PubMed1989

Two arylamine N-acetyltransferases from chicken pineal gland as identified by cDNA cloning.

A cDNA library prepared from the poly(A)-rich RNA of the chicken pineal gland obtained at night was screened with the 32P-labeled cDNA of arylamine N-acetyltransferase from the chicken liver recently isolated in this laboratory. Two positive clones (p-NAT-3 and p-NAT-10) that cross-hybridized with the liver cDNA were isolated. The cDNAs did not cross-hybridize each other under a high stringency, indicating that they corresponded to different mRNAs. When the cDNAs were inserted into an expression vector pcDL1 under the control of the early promoter of simian virus 40 and introduced into Chinese hamster ovary cells, both cDNAs expressed arylamine N-acetyltransferase activity in the transfected cells. The nucleotide sequences of the cDNAs were determined, from which amino acid sequences were deduced. Both cDNAs coded for 290 amino acids. Similarities in amino acid sequences were about 60% between p-NAT-3, p-NAT-10 and liver N-acetyltransferases. Poly(A)-rich RNA blot hybridization analysis indicated that p-NAT-3 cDNA detected a 2.2-kb band with the poly(A)-rich RNAs from the brain, gut and, less intensively, spleen, liver and kidney, while p-NAT-10 cDNA hybridized only with the poly(A)-rich RNA from the kidney. Neither cDNA detected any hybridization band with the poly(A)-rich RNA from the pineal gland, suggesting that the contents were low. Genomic Southern blot hybridization analysis showed that p-NAT-3, p-NAT-10 and liver N-acetyltransferases were encoded in a separate single gene. The properties of the enzymes expressed in the transfected cells were compared with N-acetyltransferases from the pineal gland, brain and kidney. On a DEAE-cellulose column, the kidney and p-NAT-10 enzymes appeared in the effluent fraction, whereas the brain and p-NAT-3 enzymes were eluted from the column with a gradient elution at 0.08 M NaCl. The supernatant of the pineal gland obtained in the daytime showed two peaks appearing in the effluent fraction and the eluate fraction at 0.08 M NaCl. The substrate specificity of these enzymes were examined with p-phenetidine, 2-aminofluorene, tryptamine and phenylethylamine as substrates. All the enzymes preferred arylamines to arylalkylamines, indicating that both p-NAT-3 and p-NAT-10 cDNAs encoded arylamine N-acetyltransferases.

Acetyltransferases↗

PubMed1989

Angiotensin-converting enzyme inhibitors: synthesis and structure-activity relationships of potent N-benzyloxycarbonyl tripeptide inhibitors.

A new series of gamma-D-Glu-containing N-benzyloxycarbonyl (Z) tripeptide inhibitors of angiotensin-converting enzyme (ACE) was synthesized. The effect of varying the antepenultimate amino acid residue in this series on the biological activity was studied. Introduction of Lys and Orn residues at the P1 position provided the most potent inhibitors, 25a and 25b (IC50: 3.5 and 4.9 x 10(-9) M, respectively), which exhibited an oral antihypertensive activity. This result suggests that basic amino acid residues at the P1 position play an important role in binding with the S1 subsite of ACE in this series. Oral antihypertensive activity of selected compounds was evaluated.

Angiotensin-Converting Enzyme Inhibitors↗

PubMed1989

[Correlation of serum gamma-seminoprotein and serum prostate-specific antigen in patients with prostate cancer].

Enzyme immunoassays (EIAs) using rabbit anti-gamma-seminoprotein (gamma-Sm) antibody and rabbit anti-prostate-specific antigen antibody (PA) for measurements of gamma-Sm and PA were developed in order to determine the correlation between serum gamma-Sm levels and serum PA levels in patients with prostate cancer. Each EIA for measurement of gamma-Sm or PA was revealed to be sensitive and reproducible by fundamental analysis of the quality of EIAs. A significant correlation was established between serum gamma-Sm levels and serum PA levels which were measured by these assays and expressed as protein concentration of seminal plasma (r = 0.99). This result coincided with our previous observation that gamma-Sm and PA are immunologically identical materials. We conclude, therefore, that gamma-Sm and PA should be regarded as tumor markers identical to each other for serodiagnosis of prostate cancer.

Antigens, Neoplasm↗

PubMed1989

[Analysis of antigens recognized by anti-gamma-seminoprotein antibody and anti-prostate-specific antigen antibody by means of Ouchterlony method and western blotting method].

Antigens recognized by anti-gamma-seminoprotein (gamma-Sm) antibody and anti-prostate-specific antigen (PA) antibody were analyzed immunologically by Ouchterlony method and Western blotting method. The double immunodiffusion test demonstrated that anti-gamma-Sm antibody and anti-PA antibody reacted against an identical antigen in seminal fluid or prostate homogenate. In the immunoblotting examination, anti-gamma-Sm antibody and anti-PA antibody formed a single and identical band, which gave a molecular weight of 33,000, in seminal fluid and prostate homogenate. This study demonstrated that anti-gamma-Sm antibody and anti-PA antibody recognized a single and identical antigen in seminal fluid and prostate homogenate and suggested that an identical material could be measured in sera of patients with prostate cancer by immunoassay systems using anti-gamma-Sm antibody and anti-PA antibody.

Antigens, Neoplasm↗

PubMed1989

[Evaluation of plasma PIVKA-II as a new marker for hepatocellular carcinoma].

We have measured the plasma PIVKA-II levels in 188 cases of various liver disease with HCC and malignant diseases in other organs by an EIA, using a monoclonal antibody (E-1023 kit, Eisai), and also have measured the plasma vitamin K levels in cases of HCC and cholestasis by an HPLC. Plasma PIVKA-II was detected in many cases of HCC (67%, 35 of 52 cases) and cholestasis (60%, 6 of 10 cases). In contrast, the positivities of PIVKA-II in the other diseases including benign liver diseases were very low. Combination assays of PIVKA-II and vitamin K revealed that PIVKA-II correlates with vitamin K in cholestasis but not in HCC, suggesting that PIVKA-II in HCC does not depend on a systemic deficiency of vitamin K. From these results, it was concluded that PIVKA-II is a reliable marker which can reflect the clinical course of HCC.

Adult↗

PubMed1989

[A case of localized amyloidosis of the bladder manifesting post-renal failure].

We report a case of primary localized amyloidosis of the bladder which manifested post-renal failure. A 79-year-old woman with diabetes mellitus complained of anorexia and oliguria. Computed tomographic (CT) scan showed bilateral hydronephrosis. Cystoscopic examination revealed a broad-based nonpapillary tumor in the trigonum of the bladder and CT scan demonstrated thickening of the posterior wall of the bladder. Pathological examination of the transurethral biopsy specimen revealed amyloid deposits in the submucosa, but no malignant changes were found. Cytodiagnosis of washing fluid of the bladder revealed amyloid deposits around the exfoliative cells. Serum electrophoresis showed a normal pattern. Urinary Bence-Jones protein was not detected. Amyloid deposits were not found in rectal mucosa. Systemic or secondary amyloidosis was ruled out from these findings, and primary localized amyloidosis of the bladder was diagnosed. The mass of the bladder was transurethrally resected and pig-tail stents were indwelt. These procedures gave a satisfactory result.

Acute Kidney Injury↗

PubMed1988

Arylamine N-acetyltransferase from chicken liver. I. Monoclonal antibodies, immunoaffinity purification, and amino acid sequences.

Five monoclonal antibodies against arylamine acetyltransferase (EC 2.3.1.5) from the chicken liver were established by immunizing a mouse with a partially purified enzyme preparation. None of the antibodies cross-reacted with arylamine N-acetyltransferase from the livers of cow, rabbit, and rat, nor with arylalkylamine N-acetyltransferase from the chicken pineal gland, indicating a high specificity of the antibodies. By using the antibodies, two immunoaffinity purification procedures were elaborated: A partially purified enzyme preparation was incubated with the monoclonal antibody, and the resulting enzyme-IgG complex was separated by a protein A-Sepharose column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single protein band with a molecular mass of 34 kDa in addition to the heavy and light chains of IgG. Secondly, an immunoaffinity column was prepared by immobilizing a monoclonal antibody to Sepharose 4B. After a partially purified enzyme preparation was absorbed on the column, N-acetyltransferase activity was eluted with 1 M NaCl and 1 M urea. The eluted sample contained a single 34-kDa protein. The purified enzyme preferred arylamines to arylalkylamines as substrates, indicating that it was arylamine N-acetyltransferase. The purified protein was subjected to digestion by lysylendopeptidase and separated by high performance liquid chromatography. Partial amino acid sequences of three peptides were determined by a gas-phase sequence analyzer.

Acetyl-CoA C-Acetyltransferase↗

PubMed1988

Arylamine N-acetyltransferase from chicken liver II. Cloning of cDNA and expression in Chinese hamster ovary cells.

A cDNA clone encoding the full coding region of chicken liver arylamine acetyltransferase (EC 2.3.1.5) was isolated from lambda gt10 cDNA library by screening with the 32P-labeled oligonucleotide deduced from the amino acid sequence reported in the preceding paper. The complete nucleotide sequence of the cDNA was determined, from which the amino acid sequence of N-acetyltransferase was deduced. It consisted of 1,302 nucleotides including a 861-nucleotide region coding for 287 amino acids with a molecular weight of 32,914. Two methods were applied to confirm that the cDNA encoded arylamine N-acetyltransferase. First, mRNA was synthesized in vitro by Bluescript and subsequently translated in vitro. The molecular mass of the translation product was 34 kDa consistent with that of the enzyme purified from the chicken liver. The translated protein was immunoprecipitated by a monoclonal antibody to N-acetyltransferase. Second, the cDNA was inserted into an expression vector pcDL1 and introduced into Chinese hamster ovary cells. The supernatant of the homogenate of transfected cells showed a high level of N-acetyltransferase activity with a substrate specificity comparable to that of the liver enzyme. Northern blot analysis revealed three mRNAs corresponding to 4.7, 2.0, and 1.4 kilobases. The levels of N-acetyltransferase mRNAs were relatively high in the liver and distributed in various tissues. Genomic Southern blot analysis indicated the presence of only one gene in the chicken haploid genome.

Acetyl-CoA C-Acetyltransferase↗

PubMed1988

In situ hybridization demonstrating coexpression of urotensins I, II-alpha, and II-gamma in the caudal neurosecretory neurons of the carp, Cyprinus carpio.

In order to examine the coexpression of different urotensins in the same caudal neurosecretory neurons, in situ hybridization with synthetic oligonucleotide probes specific for mRNAs for urotensins I, II-alpha, and II-gamma (UI, UII-alpha, and UII-gamma) was applied to the caudal neurosecretory system of the carp, Cyprinus carpio, together with immunohistochemistry. Almost all identifiable caudal neurosecretory neurons were labeled with the oligonucleotide and cDNA probes for UI mRNA. Further, any two of the probes for UI, UII-alpha, and UII-gamma mRNAs labeled the same neurons in serial sections. These results suggest that essentially all caudal neurosecretory neurons of the carp coexpress UI, UII-alpha, and UII-gamma. Although most neurons were densely labeled with oligonucleotide probes, only a small portion of the neurons were intensely immunoreactive. This suggests that most caudal neurosecretory neurons actively synthesize all three hormones, but that in some neurons, all or some of the hormones are rapidly transported to the urophysis, resulting in low or undetectable immunoreactivity in the perikarya.

Animals↗

PubMed1988

[A case of female urethral diverticulum calculi].

A 53-year-old nulliparous woman complained of terminal pain on urination and bended micturition. Previously, she had been treated unsuccessfully with antibiotics for chronic cystitis. On examination, a mass of approximately a thumb's head-size, was palpated at the left portion of urethral meatus. Plain film of the pelvis revealed stones under the margin of symphysis pubis. Positive pressure urethrography revealed a diverticulum slightly larger than the stones. Transvestibular diverticulectomy and urethrolithotomy were performed without any postoperative complications. In Japan, 38 cases of diverticulum calculi of the female urethra have been reported.

Diverticulum↗

PubMed1987

Monoclonal antibodies to monkey brain choline acetyltransferase: production and immunohistochemistry.

Two monoclonal antibodies to monkey brain choline acetyltransferase (ChAT) were obtained. Immunoblot analysis indicated that both antibodies revealed two adjacent protein bands on a sodium dodecyl sulfate-polyacrylamide gel electrophoresis. ChAT was demonstrated immunohistochemically by one of the antibodies in the motoneurons in the monkey brainstem and spinal cord. This antibody also revealed ChAT-positive terminal-like structures in the neuropils of lamina IX of the cervical spinal cord and interpeduncular nucleus.

Animals↗

PubMed1987

Molecular cloning and nucleotide sequence of cDNA encoding hydroxyindole O-methyltransferase of bovine pineal glands.

Messenger RNA for hydroxyindole O-methyltransferase (EC 2.1.1.4) was partially purified from poly(A)+ RNA isolated from bovine pineal glands by sucrose density gradient centrifugation. The enriched mRNA was used to prepare a cDNA library by use of expression vector lambda gt11. The library was screened with monoclonal antibodies to the enzyme, and three cDNA clones were isolated. These cloned cDNAs cross-hybridized with one another, and their fusion proteins reacted to the monoclonal antibodies with different binding properties. Hydroxyindole O-methyltransferase enzymatic activity was demonstrated in the bacteria lysate infected with lambda HIOMT-A16, the clone that contained the longest insert. An almost full-length cDNA clone was isolated from lambda gt10 cDNA library by use of the lambda HIOMT-A16 cDNA as a probe. The primary structure of hydroxyindole O-methyltransferase was determined by analyzing the nucleotide sequence of the cDNAs. It consisted of 1939 nucleotides including a 1050-nucleotide region coding for 350 amino acids. RNA transfer blot analysis indicated that mRNA encoding hydroxyindole O-methyltransferase was present only in the pineal gland and not in the brain, retina, and liver of cow.

Acetylserotonin O-Methyltransferase↗

PubMed1987

Expression and characterization of hydroxyindole O-methyltransferase from a cloned cDNA in Chinese hamster ovary cells.

We have used two kinds of expression systems to test whether the cloned cDNA encoding hydroxyindole O-methyltransferase (HIOMT) of the bovine pineal gland was functional or not. First, when mRNA was synthesized in vitro by the SP6 system and injected into Xenopus oocytes, the enzymatic activity was expressed in the oocytes. Second, the cloned cDNA was recombined to a vector under the control of the simian virus 40 early promoter and transfected to Chinese hamster ovary (CHO) cells. The enzymatic activity of the crude supernatant of transfected cells (CHO-HT2) reached to 400 pmol melatonin formed per min per mg of protein, which value was approximately 9% of that of bovine pineal supernatant. The amounts of enzyme protein estimated by immunoblotting were proportional to the enzymatic activity in both CHO and pineal gland. The content of HIOMT protein was 8- to 30-fold larger in pineal gland compared to CHO cells. On the other hand, the content of mRNA encoding the enzyme measured by dot hybridization with [32P]cDNA, was in the same range in both CHO cells and pineal glands. These data suggest that the 11-fold higher enzymatic activity in pineal gland is due to an accumulation of the enzyme protein, not to a high level of the mRNA and also indicate that the cloned cDNA can express an intact hydroxyindole O-methyltransferase enzyme in CHO cells.

Acetylserotonin O-Methyltransferase↗

PubMed1987

Glutamate receptors expressed in Xenopus oocyte by messenger RNA from invertebrate muscle.

Glutamate induced responses in the surface membrane of Xenopus oocyte by injection with mRNA from crustacea and insect muscle. Glutamate (10(-5) to 10(-4) M) elicited smooth current responses which resembled those induced by kainate. This smooth current reversed direction at about 0 mV and was blocked by a spider toxin (JSTX). Higher concentrations (more than 1 x 10(-3) M) of glutamate induced JSTX-insensitive oscillatory current responses which may be caused by Cl- activation.

Animals↗