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Biomedical subjects

T Katayama

Publications and source records attributed to T Katayama.

At least 163 records · Page 9Linked to original sources

[Rapid detection of acid-fast bacilli with Mycobacteria Growth Indicator Tube (MGIT)].

The BBL MGIT Mycobacteria Growth Indicator Tube is a novel broth based culture system for the detection of mycobacteria from clinical specimens. The tubes consist of a fluorescent indicator embedded in silicone on the bottom of a 16x100 mm round-bottom tube, filled with 4ml of an enriched BBL Middlebrook 7H9 broth base, with 0.25% glycerol. Actively growing mycobacteria consume the oxygen dissolved in the medium and fluorescence will occur when the tubes are observed with a 365nm transilluminator. The purpose of this study is to evaluate comparatively MGIT with 1% Ogawa egg medium by using two hundred and forty-five clinical specimens. The samples were digested, decontaminated and concentrated for culture using N-acetyl-L-cysteine-sodium hydroxide method. Fifty-nine of 245 (24%) clinical samples were culture positive for mycobacteria (43 M. tuberculosis complex, 12 M. avium complex and 4 other species) by one or both test systems. The MGIT detected 4 isolates of M. tuberculosis complex and 6 isolates of M. avium complex not recovered by the Ogawa egg medium, respectively. The mean time of detection of M. tuberculosis complex was 13 days (range: 2-26 days) and 19 days (range: 8-31 days) for MGIT and Ogawa egg medium, respectively, and that of M. avium complex was 5 days (range: 2-8 days) and 16 days (range: 6-22 days) for the MGIT and Ogawa egg medium, respectively. Overall, the MGIT is a sensitive culture system for the detection of mycobacteria from clinical specimens, is easy to use and may be applicable to clinical laboratories.

Culture Media↗

[Detection of Mycobacterium tuberculosis in clinical specimens other than sputum by the Mycobacterium Tuberculosis Direct Test (MTD)--assessment of sample preparation methods and clinical evaluation].

The Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test (MTD) has been widely used as a rapid test for the identification of Mycobacterium tuberculosis complex in clinical samples, and several research groups have verified its clinical usefulness. However, most of the specimens they tested were sputum, and there have been few reports on other specimens. In particular, there have been no reports on assessments of methods of preparing samples other than sputum for the MTD. We assessed methods of preparing samples other than sputum and the influence of a local anesthetic and an anticoagulant that may be present in samples, and also evaluated the MTD as a means of detecting M. tuberculosis in pleural fluid, bronchial lavage cerebrospinal fluid, urine and ascitic fluid. 1. Assessment of three sample preparation methods, i.e., the NALC-NaOH method GuSCN-Diatom nucleic acid extraction method, and the ultrasonication method, revealed that the combination of the NALC-NaOH method and the ultrasonication method, widely used to prepare sputum samples, is also a valid method of preparing other samples. 2. The local anesthetic and the anticoagulant used clinically and remained in specimens did not affect the results of the MTD. 3. Seven (36.8%) of 19 pleural fluid samples from patients diagnosed as tuberculous pleurisy were positive of M. tuberculosis by the MTD, while five (27.8%) of 18 pleural fluid samples cultured for bacteria were positive for M. tuberculosis complex. None of the 20 pleural fluid samples from patients diagnosed as non-tuberculous pleurisy were positive for M. tuberculosis complex either by MTD or culture. 4. Eight (32.0%) of 25 bronchial lavage samples from patients diagnosed as pulmonary tuberculosis were positive for M. tuberculosis complex by the MTD, while 3 (12.0%) were positive by culture. None of the 18 bronchial lavage samples from patients diagnosed as non-tuberculous disease were positive for M. tuberculosis complex either by the MTD or culture. Based on these results, it is concluded that the MTD is a very useful method of detecting M. tuberculosis in clinical samples other than sputum because it is more sensitive than culture on Ogawa's egg medium in detecting M. tuberculosis complex in pleural fluid samples, bronchial lavage samples, and so on, with the same preparation method as used for sputum.

Bacteriological Techniques↗

[Home oxygen therapy (HOT) in patients with pulmonary tuberculosis sequelae--comparison between patients medically treated and those surgically treated].

In Japan there are about 40,000 patients under home oxygen therapy (HOT), of whom about 30 to 40% are pulmonary tuberculosis sequelae (TBS). These patients can be divided into three groups depending on the treatments they had, Group 1: those who had medical treatments only, Group 2: those who had artificial pneumothorax, and Group 3: those who had thoracoplasties or other surgical treatments. The purpose of this study was to observe the distributions and possible differences in the survival rates among these groups. The study included 1537 patients with TBS under HOT followed at National Hospitals and Sanatoriums nationwide in Japan. In 819 patients the treatments were specified and of those 354 were in Group 1, 29 in Group 2, and 436 in Group 3, so that the proportion of surgically treated patients in PTS was estimated between 28.4% (436/ 1537) to 53.2% (436/819). The ages at the onset of tuberculosis, at the start of HOT and the intervals in between were 36.6, 66.2 and 29.8 in Group 1, and 26.8, 65.5, and 38.1 in Group 3 respectively. Though the ages at the start of HOT were the same, those at the onset of tuberculosis were about ten years younger in Group 3 than in Group 1. Comparing Group 1 and 3, the survival rates after the initiation of HOT (Kaplan-Meier method) was better in Group 2 (surgically treated) than in Group 1 (medically treated). It is speculated that the reason could be a better preservation of the function of the remaining lung in the surgically treated and a higher incidence of obstructive impairments in the medically treated patients.

Adolescent↗

Inhibitor of protein synthesis phase-shifts the circadian oscillator and inhibits the light induced-phase shift of the melatonin rhythm in pigeon pineal cells.

Our recent study showed that dissociated pigeon pineal cells expressed a circadian oscillation of melatonin release which entrained to light-dark cycle and persisted under constant darkness in vitro, suggesting that pigeon pineal cells contain the circadian oscillator and photoreceptors. Six-hour pulses of anisomycin, an inhibitor of protein synthesis that acts at hte 80S ribosomal subunit, induced steady state and phase depended phase shifts of the circadian oscillation of melatonin release. The phase advances and delays were produced at CT 7.9 h and between CT 18.6 h and CT 4.5 h, respectively. The magnitudes of phase shifts were dose dependent and correlated with the magnitudes of inhibition of protein synthesis determined at CT 4.5 h. Furthermore, anisomycin blocked the light-induced phase advance. Two dimensional electrophoresis revealed that synthesis of two proteins with Mr of 17,600 and less than 5000 are stimulated by a 3-h light pulse at CT 18.6 h which corresponds to the light-induced phase advance region. These results suggest that 80S ribosomal protein synthesis is involved in normal or light-entrainment functions of the circadian oscillator in pigeon pineal cells.

Animals↗

Development of regulation of melatonin release in pineal cells in chick embryo.

Melatonin release in a pineal cell culture from 13- and 14-day-old chick embryos increased during the dark phase and decreased during the light phase of a 12 h light:12 h dark cycle. When the light-dark cycle was reversed, the pattern of melatonin release in the culture also reversed. 8-Bromo cyclic-AMP stimulated melatonin release in both the light and dark phases. However, no rhythm of melatonin release was detected under constant dark (DD) conditions in a cell culture from 14-day-old chick embryos. In 18-day-old chick embryos, the pineal cell culture expressed a circadian rhythm of melatonin release under DD conditions. These results indicate that mechanisms regulating melatonin synthesis in the avian pineal gland are established during embryonic life.

8-Bromo Cyclic Adenosine Monophosphate↗

Release of biological activities from quiescent fibronectin by a conformational change and limited proteolysis by matrix metalloproteinases.

We reported that specific biological activities are confined to three domains of the fibronectin (Fn) molecule [Fukai et al. (1991) J. Biol. Chem. 266, 8807; Fukai et al. (1993) Biochemistry 32, 5746]: the potent ability to stimulate the adipocyte differentiation of ST-13 cells is in the amino-terminal fibrin-binding (Fib 1) domain (referred to as Fib 1 domain activity); the RGD-dependent activities that stimulate NIH-L13 cell migration and inhibit adipocyte differentiation are in the central cell-binding (Cell) domain (Cell domain activity); and the activity that stimulates cell migration in a RGD-independent manner is in the carboxyl-terminal fibrin-binding (Fib 2) domain (Fib 2 domain activity). Human plasma Fn which was purified without exposure to a denaturant, such as urea, exhibited no Fib 1, Fib 2, or Cell domain activity. By exposure to urea or surface adsorption, Fn showed Cell domain activity but not those of the Fib 1 and Fib 2 domains. Whether the cryptic domain activities are disclosed or not depended on whether or not the responsible domains were irreversibly exposed from confined environments of Fn structure as confirmed by light-scattering measurement and enzyme immunoassay using domain-specific monoclonal antibodies. We then investigated the action of matrix metalloproteinases (MMPs) in liberating the Fib 1, Cell, and Fib 2 domain activities. Matrilysin released only the Cell domain activity. In contrast, stromelysin, collagenase, and especially gelatinase A additionally liberated the Fib 1 and Fib 2 domain activities. The Fib 1, Fib 2, and Cell domains acquired much higher activities when they were freed from linkage with adjacent domains.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

Isolation and characterization of inhibitory factors of DNA polymerase III holoenzyme from Escherichia coli.

We isolated fractions by Mono Q chromatography that inhibited the activity of Escherichia coli DNA polymerase III holoenzyme using an assay system with a primed single-stranded DNA template coated with single-stranded DNA binding protein (SSB). The inhibitory activities were inactivated by heat-treatment at 100 degrees C for 10 min, suggesting that they are proteins. The factors did not inhibit the activity of RNA polymerase of Escherichia coli. The inhibitory effects were less potent for the activities of the large (Klenow) fragment of DNA polymerase I and T4 DNA polymerase than for DNA polymerase III holoenzyme. No degradation of single- or double-stranded DNA was observed in the fractions, indicating that inhibition was not due to degradation of the DNA.

DNA↗

Sequence analysis of the MAGE gene family encoding human tumor-rejection antigens.

The MAGE multigene family, which includes the MAGE-1 and -3 genes that encode tumor-rejection antigens on HLA-A1 recognized by cytotoxic T-lymphocytes (CTL), is preferentially expressed at the mRNA level on human malignant cells, but not on normal cells. However, little is known about the MAGE-4, -41 and -6 genes. In this study, we have amplified 1040 bp (MAGE-1), 1061 bp (MAGE-3 and -6) and 1064 bp (MAGE-4 and -41) cDNA fragments, including the entire coding sequences (927-951 bp), using the reverse transcription-polymerase chain reaction (RT-PCR) method followed by nucleotide (nt) sequencing. One member had greater than 80 or 66% homology with the other members at the nt or deduced amino acid (aa) levels, respectively. Higher homology was found between MAGE-3 and -6 (98% at the nt level) and also between MAGE-4 and -41 (98%). The results of this investigation demonstrated high homology, as well as the clear differences between the members of the MAGE family at the coding sequence level.

Amino Acid Sequence↗

Effects of benzoic acid and its analogues on insulin and glucagon secretion in sheep.

The effects of benzoic acid and its analogues on insulin and glucagon secretion were investigated in conscious sheep. Intravenous injections of benzoic acid increased plasma insulin and glucagon concentrations in a dose-dependent manner between 39-1250 mumol/kg, with ED50s for increasing both hormones of about 625 mumol/kg. Various derivatives of benzoic acid (625 mumol/kg) were administered and structure-activity relationships were examined. A single carboxylic group was essential for stimulating insulin and glucagon secretion, since both hormone responses were abolished with compounds in which the carboxylic group was replaced by sulfonic or phosphoric groups, or in which another carboxylic element was introduced (phthalic acids). Most of the compounds which introduced other elements (amino and hydroxy groups, and halogens) onto the benzene ring had an altered stimulating activity. Thus the pancreatic endocrine system can recognize the chemical structure of benzoic acid and its derivatives in detail and induce insulin and glucagon secretion in sheep.

Animals↗

DnaA protein is sensitive to a soluble factor and is specifically inactivated for initiation of in vitro replication of the Escherichia coli minichromosome.

DnaA protein loses the capacity to initiate chromosomal replication when treated with a soluble cell extract. This inactivation depends upon DNA and hydrolyzable ribonucleoside triphosphate. The extract does not affect the activities of other replicative proteins or the ability of DnaA to initiate replication of single-stranded DNA that contains a DnaA-binding hairpin, indicating that the inhibitory effect is specific for the action of DnaA at oriC. Gel filtration experiments implicate a 150-kDa factor as being responsible. Mutant DnaAcos protein, which causes overinitiation in vivo, is insensitive to the inactivating factor, suggesting a requirement for this negative control in vivo. We propose that a soluble factor controls initiation through down-regulation of DnaA protein.

Adenosine Triphosphate↗

Modified one-layer microsurgical vasovasostomy in vasectomized patients.

Bilateral vasovasostomies were performed in 7 previously vasectomized patients. The method employed was a modification of a one-layer anastomotic technique. There was no correlation between the presence of absence of sperm in the vas fluid, presence or absence of sperm granuloma, and site of vasectomy. No correlation was revealed between the presence or absence of sperm in the vas fluid and the duration of obstructive period. After vasovasostomy, sperm was observed in the ejaculate in 86% of the patients. Only one patient's partner became pregnant. However, the partner of one patient with short postoperative period and good seminal finding after vasovasostomy was expected to become pregnant. This modified method of one-layer microsurgical vasovasostomy can be performed more easily and quickly, but requires further clinical experience and evaluation of usefulness.

Adult↗

Clinical study on prognosis of metastatic prostate cancer based on extent of disease on pretreatment bone scintigraphy.

The pretreatment bone scintigrams of 58 patients with prostate cancer with bone metastasis were reviewed and the prognostic value of the initial extent of bone involvement was compared with that of other pretreatment characteristics. The extent of bone metastasis revealed by the scan was related to survival. The serum level of alkaline phosphatase at pretreatment and pathological grade also had some predictive value. Although the pathological grade of primary tumours was related to prognosis, the survival of patients with the same histological grade differed according to the initial extent of disease; patients with extensive disease along with raised serum alkaline phosphatase (twice or more as high as the cut-off value) had poorer prognosis than did those with lower alkaline phosphatase or smaller extent of disease. The extent of bone involvement in combination with serum alkaline phosphatase level therefore apparently has higher prognostic value than does disease extent alone.

Acid Phosphatase↗

Protein kinase C inhibitor potentiates the agonist-induced GTPase activity in COS cell membranes expressing delta-opioid receptor.

In COS-7 cell membranes expressing cloned delta-opioid receptor, [D-Ser2, Leu5]enkephalin-Thr6, an opioid delta-agonist, showed no significant stimulation of high-affinity GTPase, while this agonist binding showed a guanine nucleotide sensitivity. Significant stimulation of GTPase activity by this agonist was observed only when the cells were pretreated with 0.1 microM calphostin C, a protein kinase C inhibitor, and when this inhibitor was further added to the reaction mixture at 1 microM. These findings suggest that protein kinase C is involved in the heterologous desensitization of delta-opioid receptor in the cells.

Analgesics↗

A novel proteinase, glucagon-degrading enzyme, secreted by a human pancreatic cancer cell line, HPC-YO.

Sixty-four kinds of cell lines were examined as to their ability to degrade glucagon using conditioned-media obtained from their protein-free cultures. Two human tumor cell lines were shown to produce this activity, and the cell line, HPC-YO, established from a human pancreatic carcinoma was shown to produce the highest level of activity. The glucagon-degrading enzyme (GDE) was purified from HPC-YO conditioned-medium by a combination of ion-exchange, gel filtration, and hydroxylapatite column chromatographies. The purified GDE also degraded vasoactive intestinal polypeptide (VIP) and secretin, however, it did not cleave EGF, gastrin, insulin, somatostatin, substance P, neurotensin, or growth hormone. The molecular weight of GDE is 83,000, as determined on SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of GDE was blocked, and the five partial amino acid sequences obtained on lysyl-endopeptidase digestion were determined to be N-L-T-E-E-Y-D-V-S-D-G-E-I-E-L-L-Y-E-K, V-E-T-Y-Y-D-L-L-F-E-K, L-Y-W-F-L-D-E-A-K, S-N-S-T-S-Y-V-K, and Y-Y-A-S-T-S-Y-D-D-T-Y-K. The same or homologous amino acid sequences have not been found in known proteins, demonstrating that GDE is a novel peptidase that degrades the secretin family: glucagon, VIP, and secretin.

Amino Acid Sequence↗

Characterization of Escherichia coli DnaAcos protein in replication systems reconstituted with highly purified proteins.

Excessive initiation of chromosomal replication occurs in the dnaAcos mutant at 30 degrees C. Whereas purified wild-type DnA protein binds ATP and ADP tightly, DnaAcos protein is defective for such nucleotide binding. As initiation is a multistep reaction and DnaA protein functions at each step, activities of DnaAcos protein need to be examined precisely. DnaAcos protein specifically bound a DNA fragment containing the chromosomal replication origin with an affinity similar to that seen with the wild-type protein. In a system reconstituted with purified proteins at 30 degrees C, the mutant protein initiated replication of single-stranded DNA that contains a DnA-binding hairpin structure. Thus, DnaAcos protein basically sustains affinity to a DnaA-binding sequence and functions in the loading of DnaB helicase onto single-stranded DNA. Thermal stabilities of wild-type DnA and DnaAcos activities were comparable. Unlike wild-type DnaA protein, DnaAcos protein was inactive for minichromosomal replication in systems reconstituted with purified proteins in which the ATP-bound form of DnaA protein is required for initiation. Taken together, the data indicate that the prominent defect in DnaAcos protein appears to be the inability to bind nucleotide.

Bacterial Proteins↗

Specific amplification of Rickettsia japonica DNA from clinical specimens by PCR.

The gene encoding the 17,000-molecular-weight genus-common antigen (17K genus-common antigen) has been cloned and sequenced from Rickettsia japonica. The primer pair used for PCR was designed from this sequence. A 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and also the DNA from blood clots of patients with spotted fever group rickettsiosis. The results indicated that this method is suitable for the diagnosis of spotted fever group rickettsiosis in Japan.

Antibodies, Bacterial↗

Immunopathological mechanisms of human T cell lymphotropic virus type 1 (HTLV-I) uveitis. Detection of HTLV-I-infected T cells in the eye and their constitutive cytokine production.

The immunopathology of human T cell lymphotropic virus type 1 (HTLV-I) uveitis was addressed by using T cell clones (TCC) established from the intraocular fluid of patients with HTLV-I uveitis. Proviral DNA of HTLV-I was identified in 55 out of 94 (59%) or 13 out of 36 (36%) TCC from the ocular fluid or the peripheral blood of these patients, respectively. Most of HTLV-I-infected TCC had a CD3+ CD4+ CD8- phenotype. HTLV-I infection on TCC was confirmed by analysis of the viral mRNA, nucleotide sequence, virus-associated proteins, and virus particles. HTLV-I-infected TCC, but not HTLV-I negative TCC, constitutively produced high amounts of IL-6 (1,336 +/- 1,050 pg/ml) and TNF-alpha (289 +/- 237 pg/ml) in the absence of any stimuli. HTLV-I-infected TCC from the ocular lesion also constitutively produced high amounts of IL-1 alpha (12,699 pg/ml), IL-2 (61 pg/ml), IL-3 (428 pg/ml), IL-8 (1,268 pg/ml), IL-10 (28 pg/ml), IFN-gamma (5,095 pg/ml), and GM-CSF (2,886 pg/ml). Hydrocortisone, a drug effective in vivo for the treatment of HTLV-I uveitis, severely depressed cytokine production in vitro in most cases. In summary, the results demonstrated direct evidence of HTLV-I infection of the eye and suggest that cytokines produced by HTLV-I-infected T cells are responsible for the intraocular inflammation in patients with HTLV-I uveitis.

Adult↗

Cytogenetical studies on the genus Oryza. XIV. Intergeneric hybridizations between tetraploid Oryza species and diploid Leersia species.

Intergeneric hybrids involving Oryza punctata (2n = 48, BBCC) with Leersia tisseranti (2n = 24), O. punctata with L. perrieri (2n = 24), O. latifolia (2n = 48, CCDD) with L. tisseranti and O. latifolia with L. perrieri were produced at frequencies varying from 0.11% to 0.23% of the pollinated spikelets. Plant morphologies of the hybrids strongly resembled the tetraploid Oryza species. Five hybrids obtained from the four cross-combinations had the expected chromosome number of 2n = 36 (trihaploid) in the somatic cells. The average meiotic chromosome pairings per cell were 0.23II + 35.58I + 0.14(1)/2I (dividing univalent) for O. punctata x L. tisseranti; 0.11II + 35.51I + 0.22(1)/2I for O. punctata x L. perrieri; 0.17II + 35.63I + 0.11(1)/2I for O. latifolia x L. tisseranti; and 0.25II + 35.49I + 0.15(1)/2I for O. latifolia x L. perrieri. From these results, it seems that most of the bivalents observed at MI with a low frequency had originated from the autosyndetic association of the chromosomes of O. punctata or O. latifolia. The results described above suggest that there is no genomic relationship between the parental species in each cross combination.

Chromosomes↗