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Biomedical subjects

M Takasugi

Publications and source records attributed to M Takasugi.

At least 19 recordsLinked to original sources
PubMed1992

Reduction and inactivation of the sarcoplasmic Ca(2+)-ATPase by 2-mercaptoethanol--contrast to the (Na+,K+) ATPase.

The sarcoplasmic Ca(2+)-ATPase was reduced with 300 mM 2-mercaptoethanol at elevated temperatures (40-45 degrees C) with a concomitant loss of ATPase activity. The reduction and inactivation of the Ca(2+)-ATPase proceeded rapidly in the absence of Ca2+. The Ca(2+)-ATPase was also inactivated with 2-mercaptoethanol in the presence of diluted SDS (0.4 mg/ml) even at 20 degrees C. In contrast to the (Na+, K+) ATPase, the inactivated Ca(2+)-ATPase in the presence of diluted SDS was sedimented by the centrifugation at 100,000 x g for 30 min.

Animals↗
PubMed1992

[Studies on purine-pyrimidine metabolism (1)--Quantitation of purine-pyrimidine metabolites and allopurinol-oxipurinol in biological fluids].

A reversed-phase high-performance liquid-chromatography method for determining simultaneous quantitation of purine-pyrimidine metabolites, allopurinol and oxipurinol in plasma and urine samples was studied. Separation was optimal with phosphate buffer (10 mmol/l, pH 5.0) containing 1% methanol as an eluent and mu Bondapak C18 as a column. An isocratic separation of a standard mixture of 13 compounds was achieved within 40 minutes with adequate reproducibilities (coefficient of variation: 2.49% for 1.63 mumol/l orotidin-0.12% for 50 mumol/l uridine). A simple ultrafiltration of plasma yielded quantitative recoveries (uric acid: 101.7-107.5%, hypoxanthine: 90.4-102.8%, xanthine: 95.9-99.5%, oxipurinol: 104.4-107.1%, allopurinol: 97.4-103.4%). Compounds were identified by their retention times, absorbance ratios, co-elution with standards and enzymic shifts. In addition to the above compounds, simultaneous quantitation of pseudouridine, uridine, adenine and inosine in the plasma would be possible under the same conditions.

Allopurinol↗
PubMed1992

[Dynamic process of colonization by Haemophilus influenzae in children--is H. influenzae normal flora in the throat?].

Eighty-seven oropharyngeal isolates of Haemophilus influenzae were obtained by two time cultures six months apart from a total of 288 children who attend a kindergarten. We analyzed the strains by comparing their serotypes, biotypes, beta-lactamase activity and by performing electrophoresis of outer membrane proteins on polyacrylamide gels. Only nineteen strains were not identical, the rest of the 68 strains were classified into 23 types. During 6 months at least 15 types of strains lost from this group and 21 types of new strains were obtained. There were no children who had identical pairs of H. influenzae in their oropharynx during the 6 months. The classification of strains in oropharynx suggested that person-to-person transmission of nontypable H. influenzae can occur. We concluded that oropharyngeal colonization by nontypable H. influenzae is not a normal flora in children.

Bacterial Outer Membrane Proteins↗
PubMed1992

Expression and assembly of Torpedo californica (Na,K)ATPase alpha-subunit truncated at N-terminal end in Xenopus oocytes.

cDNAs for mutant alpha-subunits of Torpedo californica (Na,K)ATPase variously truncated at the N-terminal end were constructed and transcribed in vitro. Each of the mRNAs thus synthesized was co-injected into Xenopus oocytes together with mRNA for wild-type beta-subunit. Truncation of the alpha-subunit at trypsin accessible site T2(removal of the N-terminal 36 residues, alpha delta K37) led to a decrease in ouabain-sensitive ATPase activity and ouabain-binding capacity, leaving the amount of immunoprecipitable alpha-subunit unchanged. The Km values for Na+ and K+ of alpha delta K37 were about 10mM and 2mM, respectively, and fall in the same range for the wild-type ATPase. Truncation of the alpha-subunit leaving lysine-54(alpha delta K54) or alanine-79(alpha delta A79) resulted in the loss of the ATPase activity as well as a substantial decrease in the amount of immunoprecipitable alpha-subunit. Since the beta-subunit assembles with and thereby stabilizes the alpha-subunit, which is otherwise degraded rapidly, these results suggest that the segment of the alpha-subunit between lysine-37 and lysine-54 is involved in the assembly with the beta-subunit leading to the formation of the stable and active alpha beta complex.

Animals↗
PubMed1992

[A case of hyperparathyroid bone disease and the review of renal osteodystrophy].

We report here a typical case of hyperparathyroid bone disease associated with CRF on maintenance HD and review on Renal Osteodystrophy. A 39 year-old female patient was admitted because of polyarthralgia and pruritus. She had a history of HD due to CGN for about 13 years. Laboratory data showed an increase in serum PTH and Alkaline phosphatase level. The evidence of osteitis fibrosa was revealed by bone Xp and scintigraphy. Enlarged solid masses were found in her neck by echogram and parathyroid scintigraphy. She was diagnosed as hyperparathyroid bone disease and total parathyroidectomy c autoplantation was done. Shortly after the surgical treatment, subjective symptoms were relieved and PTH level was normalized. The bone Xp findings improved gradually.

Adult↗
PubMed1992

Comparative effects of loop diuretics on AVP-receptor binding and AVP-sensitive adenylate cyclase activity.

The effects of loop diuretics (azosemide, ethacrynic acid and furosemide) on arginine vasopressin (AVP) receptor-adenylate cyclase components were compared in rat renal basolateral membranes. AVP binding was inhibited by these loop diuretics at concentrations above 10(-4) M. At the IC50 of azosemide and ethacrynic acid, the Kd values were significantly increased, while the Bmax values remained unchanged. These findings indicate an inhibitory effect of loop diuretics at high concentrations on the AVP binding to its receptors. Both the basal (AVP-unstimulated) and AVP-stimulated cyclic AMP productions were also inhibited by addition of these drugs. The inhibitions of the AVP binding and AVP-sensitive adenylate cyclase activity were dose-dependent. The above findings suggest that loop diuretics, especially azosemide and ethacrynic acid, can inhibit the basal and AVP-sensitive adenylate cyclase activities directly and also indirectly via the AVP receptor, at least in part. Comparing the loop diuretics, azosemide exerts a similar effect to ethacrynic acid, and they have a more potent antagonistic effect than furosemide with respect to AVP adenylate cyclase activation.

Adenylyl Cyclases↗
PubMed1991

Sequence-specific photo-induced cross-linking of the two strands of double-helical DNA by a psoralen covalently linked to a triple helix-forming oligonucleotide.

On the basis of the structure of DNA-psoralen bis adducts (formed by psoralen with two thymines on opposite strands), a psoralen-oligonucleotide conjugate was designed to photoinduce a cross-link between the two DNA strands at a specific sequence. Psoralen was attached via its C-5 position to a 5'-thiophosphate group of an 11-mer homopyrimidine oligonucleotide. The 11-mer binds to an 11-base-pair homopurine.homopyrimidine sequence of a DNA fragment, where it forms a triple helix. Upon near-UV-irradiation, the two strands of DNA are crosslinked at the TpA step present at the triplex-duplex junction. The reaction is specific for the homopurine.homopyrimidine DNA sequence and requires both oligonucleotide recognition of the DNA major groove and intercalation of psoralen at the triplex-duplex junction. The yield of the photo-induced cross-linking reaction is quite high (greater than 80%). Such psoralen-oligonucleotide conjugates are probes of sequence-specific triple-helix formation and could be used to selectively control gene expression or to induce site-directed mutations.

Base Sequence↗
PubMed1991

[A case of sarcoid granulomatous interstitial nephritis improved by steroid therapy].

We report a case of sarcoid granulomatous tubulointerstitial nephritis diagnosed by renal biopsy. A 60-year-old man presented with productive cough, and exertional dyspnea of 3 months duration. A chest X-ray film revealed diffuse reticulonodular infiltrates in both lung fields. A transbronchial lung biopsy specimen showed inflammation of the alveolar septum associated with non-caseating granulomas. The patient also had tubular proteinuria and glucosuria. Ga-scintigraphy demonstrated an abnormal accumulation of gallium in both lungs and kidneys. Renal function tests revealed tubular dysfunction. Tubulointerstitial nephropathy was suspected. A renal biopsy specimen exhibited tubulointerstitial nephritis associated with numerous non-caseating granulomas, similar to the findings of the lung biopsy specimen. No glomerular abnormalities were evident. Later, a scalene node biopsy confirmed the diagnosis of sarcoidosis. Prednisolone therapy yielded a favorable outcome for both the renal and pulmonary involvement. During the corticosteroid therapy, measurement of the urinary beta-2-microglobulin concentration proved a valuable monitoring tool for assessing the recovery of the tubular impairment.

Humans↗
PubMed1990

Variations in intrathyroidal lithium content and their effect on the iodide uptake in mouse thyroid.

Lithium (Li) is accumulated in the thyroid but the mechanism of Li accumulation is not known. In the present study, the causes of variation in Li concentration in the thyroid and the relation between cellular Li and iodide were examined. This was done by using mice treated with Li (0.01% as Li2CO3) for 4 weeks, co-administered with propylthiouracil (PTU, 0.5 mg/ml daily p.o.) or thyroxine (T4, 0.5 micrograms per day i.p.) for last 10 days. The total content of Li in a whole thyroid (ng/thyroid) was not changed through treatment with PTU or T4. But the Li concentration in terms of mg/kg of the gland was reduced with PTU and was unchanged or slightly increased with T4, due to the change in the mass of thyroid. Furthermore, short-term (3 h) Li uptake in the thyroid was not affected by pre-treatment with PTU or T4. These results indicate that the variation in thyroidal Li concentration was not due to a direct effect of PTU or T4 on Li transport, but to a thyroidal condition caused by the secondary influence of drugs. A measurement of the thyroid: serum iodide concentration ratio (T/S) of 125I showed that the iodide uptake was reduced when intracellular Li concentration was high, and that PTU alone elevated the T/S, but Li + PTU brought it back to a normal level; whereas, T4 alone diminished the T/S, Li + T4 made it rise significantly more than did T4 alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

[Inhalation therapy of antibiotics].

We produced experimental murine Klebsiella pneumoniae pneumonia by air-borne infection using exposure apparatus (LD50: 9.7 X 10 C.F.U./lung). The MIC value of cefazolin was 1.56 micrograms/ml, and that of gentamicin was 0.39 micrograms/ml. We treated this murine pneumonia with aerosolized antibiotics. The infected mice received inhalation of 50 mg (5 mg/ml), 100 mg (10 mg/ml), 200 mg (20 mg/ml) and 400 mg (40 mg/ml) gentamicin were alive. Survival rate of the infected mice treated with inhalation of 1000 mg (100 mg/ml) cefazolin was low, but that of them received inhalation of 500 mg (50 mg/ml) cefazolin was high. On the basis of these experiments it is suggested that aerosol of 50 mg/ml cefazolin gets to alveoli, and inhalation therapy of low level of cephems is proper for bacterial respiratory infections. Inhalation therapy of 50 mg/ml cephem for chronic bronchitis had a marked clinical effect. This proves that aerosol of cephem gets to infected bronchi.

Administration, Inhalation↗
PubMed1989

[Respiratory tract infections caused by Branhamella catarrhalis in outpatients with pneumoconiosis].

To investigate the occurrence of Branhamella catarrhalis respiratory tract infections in 109 outpatients with pneumoconiosis, clinical and bacteriological studies were performed during a 4-year period from April 1984 to March 1988. B. catarrhalis was isolated in 26 patients; only three of these received continuous corticosteroid treatment. The incidence of B. catarrhalis respiratory tract infections increased gradually during the years 1984-1986, but decreased for the first time in 1987 compared with the previous year. There was a seasonal variation in isolations with a peak incidence during the winter, a pattern in contrast to Haemophilus influenzae. Almost all isolates produced beta-lactamase. B. catarrhalis found in mixed culture was usually in association with H. influenzae or Streptococcus pneumoniae. The isolation rates for B. catarrhalis in sputum of patients with pneumoconiosis followed those of H. influenzae and S. pneumoniae, and almost all strains were positive for beta-lactamase, so B. catarrhalis should be admitted that it is a primary pathogen.

Adult↗
PubMed1989

Effect of lithium carbonate administration singly or in combination with some psychotropic drugs on the radioiodide uptake by mouse thyroid.

The present study dealt with two objects; the first object was to examine whether or not lithium uptake in thyroid is modified by the thyroid state and how the intrathyroidal Li affects iodide uptake by the thyroid. Male and female mice were given lithium carbonate (Li) with propylthiouracil (PTU) or thyroxine (T4). Li was measured by a flameless atomic absorption spectrometry. The total Li content in thyroid was unaffected with PTU or T4, however, Li concentration per unit mass was reduced by PTU but unaffected by T4. The thyroid: serum ratio (T/S) of 125I resulted that the T/S became higher when Li concentration per unit mass was lower and vice versa, suggesting that Li uptake is controlled by thyroid states and Li in the gland interferes with the iodide uptake. Serum triiodothyronine (T3) and T4 by radioimmunoassay showed that PTU alone and in combination with Li lowered serum T4, while a high level of T4 by its supplement was suppressed by co-administration of Li. T3 level was lowered by Li alone, but not severely affected by other drugs. The results suggest that Li enhances T4 clearance without T4-T3 conversion. The second object was to examine the effect of combined psychotropic drugs on thyroid function. Carbamazepine (CBZ), haloperidol (HLP) and imipramine (IPA) were given singly or in combination with Li to examine their effects on the T/S of 125I. Only CBZ reduced the T/S but CBZ plus Li had no summative effect. Neither HLP nor IPA affected the T/S, singly or in combination with Li, suggesting that HLP or IPA does not interfere with an iodide pumping machinery. No distinct sex difference was observed in drug effects.

Animals↗
PubMed1988

Site-specific cleavage of single-stranded and double-stranded DNA sequences by oligodeoxyribonucleotides covalently linked to an intercalating agent and an EDTA-Fe chelate.

An oligodeoxythymidylate, oligo [d(T8)], was covalently linked to an acridine derivative via its 3' end and to EDTA via its 5' end. The octathymidylate was targeted to a single-stranded DNA fragment 27 nucleotides in length containing an octadeoxyadenylate sequence. In the presence of Fe(II) and a reducing agent (dithiothreitol) cleavage reactions were induced in the nucleotide sequence. The extent of the reaction was dependent on oligo concentration, salt concentration and temperature. Dissociation of the complexes at high temperature or low salt concentration abolished the site-specific cleavage reactions. Treatment of the reacted DNA with piperidine or piperidine-formiate strongly enhanced the yield of cleavage reactions demonstrating that damages were induced on nucleic acid bases by the EDTA-Fe complex covalently linked to the octathymidylate. At high salt concentration (1 M NaCl) or in the presence of spermine and ethylene-glycol a triple helix was formed involving the 27-mer DNA fragment and two oligo[d(T8)]. One of the oligo[d(T8)] was bound parallel and the other antiparallel to the oligo[d(A8)] complementary sequence. Cleavage reactions were induced on both sides of this oligo[d(A8)] target sequence. When a 27-mer duplex was used as a target the oligo[d(T8)] was bound in a parallel orientation with respect to the oligo[d(A8)]-containing strand in the major groove of the double helix. Cleavage reactions were induced on the oligo[d(A8)]-containing strand by the EDTA-Fe chelate attached to the 5' end of the oligo[d(T8)].

Acridines↗