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Biomedical subjects

Y Hashimoto

Publications and source records attributed to Y Hashimoto.

At least 829 records · Page 46Linked to original sources
PubMed1994

Purification, characterization, and crystallization of two types of lipase from Rhizopus niveus.

The purification and some properties of two types of lipase (Lipase I and Lipase II) from Rhizopus niveus are described. The enzymes were purified to homogeneity by column chromatographies on DEAE-Toyopearl (1 pass) and CM-Toyopearl (2 passes). Lipase I consists of two polypeptide chains [a small peptide with sugar moiety (A-chain) and a large peptide of molecular weight 34,000 (B-chain)]. Lipase II has a molecular weight of 30,000 consisting of a single polypeptide chain. Lipase I appeared to be converted to Lipase II by limited proteolysis by a specific protease a small amount of which is in the culture supernatant from Rh. niveus, because one of the peptides formed has the same N-terminal sequence and C-terminal amino acid as Lipase II, as well as the molecular mass estimated by SDS-PAGE. Lipase I had a pH optimum of 6.0-6.5 and a temperature optimum of 35 degrees C, while, for Lipase II these values were pH 6.0 and 40 degrees C. Both enzymes were obtained in the crystalline state using the hanging drop method of vapor diffusion and PEG as the precipitating agents.

Amino Acid Sequence↗
PubMed1994

Nitrile hydratase gene from Rhodococcus sp. N-774 requirement for its downstream region for efficient expression.

For improvement of the production of nitrile hydratase (NHase) from Rhodococcus sp. N-774 by recombinant DNA techniques, several plasmids, each of which had a deletion of the upstream or downstream region of the genes encoding the alpha and beta subunits of NHase, were constructed. Enzyme assays of recombinant R. rhodochrous and Escherichia coli cells showed that a downstream region of the NHase genes was indispensable for the production of active NHase in both cells, but for the production of the active amidase, no genes other than the amidase structural gene were required. The nucleotide sequence of the downstream region contained a single open reading frame (Orf1188) with 396 amino acids. Orf1188 showed similarity in amino acid sequence to P47K, an open reading frame found downstream of the NHase genes from Pseudomonas chlororaphis B23, and also to the cobW gene product, which may be involved in cobalamin biosynthesis in Pseudomonas denitrificans. Because the distance between the TGA stop codon for the NHase beta-subunit and the ATG codon for Orf1188 is only 98 bp, and because production of both Orf1188 and NHase is dependent on a promoter upstream of the amidase gene, these genes appear to be co-transcribed in a polycistronic manner, forming an operon. By optimization of the culture conditions of R. rhodochrous carrying pKRNH2, which contained the amidase, NHase, and Orf1188 genes, the transformant showed the NHase activity 6-fold higher than that of the original strain, Rhodococcus sp. N-774.

Amino Acid Sequence↗
PubMed1994

Alternative splicing of bovine terminal deoxynucleotidyl transferase cDNA.

Two cDNA fragments of terminal deoxynucleotidyl transferase, which contained new insertion sequences, were found from bovine thymus cDNA. One of the clones encoded 18 new amino acids and the other encoded 9 new amino acids. In the results of genomic structure analysis around the new insertion sequences, alternative splicing of the bovine terminal deoxynucleotidyl transferase gene was suggested.

Alternative Splicing↗
PubMed1994

The carcinogenicity of methoxyl derivatives of 4-aminoazobenzene: correlation between DNA adducts and genotoxicity.

To elucidate the cause of the difference in genotoxic activity between carcinogenic 3-methoxy-4-aminoazobenzene (3-MeO-AAB) and noncarcinogenic 2-methoxy-4-aminoazobenzene (2-MeO-AAB), we analyzed DNA adducts in the livers of rats exposed to either of these chemicals and studied the resulting biologic potential with the aid of in vitro modified M13 phage DNA. 32P-Postalbeling analysis revealed that the carcinogen 3-MeO-AAB produced 20-fold higher amounts of adducts than did 2-MeO-AAB. Five adducts were formed in the 3-MeO-AAB case whereas only one adduct was apparent in 2-MeO-AAB-treated rat. Studies of in vitro DNA replication using N-hydroxy (N-OH)-aminoazo dye-modified M13 phage DNA as a template demonstrated inhibition by 3-MeO-AAB adducts to be substantially greater than in the 2-MeO-AAB-adducts. The specificity of mutagenesis induced in M13mp9 phage DNA by these chemicals also was analyzed after transfection into SOS-induced Escherichia coli JM103, mutation frequencies being higher with N-OH-3-MeO-AAB- than N-OH-2-MeO-AAB-modified DNA. The mutation spectra differed in each case. Our data suggest that the difference in hepatocarcinogenic activity between the two chemicals depends not only on qualitative and quantitative variation in adduct formation but also on conformation changes in modified DNA.

Animals↗
PubMed1994

Perforin-like immunoreactivity in feline globule leukocytes and their distribution.

Distributional and immunohistochemical characteristics of the feline globule leukocyte (GL) were investigated by light microscopy. The GL, which contained eosinophilic large granules in the cytoplasm, was frequently found in the epithelia of the intestine and gall bladder, and less frequently found in those of the gastric pit, intrahepatic bile duct, and interlobular secretory duct of the pancreas. No GL was seen in the respiratory and urogenital organs. The GLs composed a homogeneous cell population including no mast cells according to the following histochemical stainings; phosphotungstic acid hematoxylin, alcian blue and peroxidase. The feline GL showed perforin-like immunoreactivity to anti-human perforin monoclonal antibody, but did not show histamine-immunoreactivity to anti-histamine polyclonal antibody. The results suggest that the feline GL is a lineage of large granular lymphocytes. The epitheliotropism and characteristics as granular lymphocytes of the feline GLs were similar to those of the intestinal gamma delta T cells of the mouse.

Animals↗
PubMed1994

Respiratory muscle strength and hemodynamics in chronic heart failure.

To examine whether respiratory muscle weakness is associated with cardiac function and/or exercise capacity in chronic heart failure (CHF), 23 patients with CHF were evaluated with respiratory muscle strength, pulmonary function tests, cardiac catheterization, and exercise test. The subjects were divided into three groups on their New York Heart Association (NYHA) functional class. Group A consisted of 13 patients with NYHA functional classification class 3 or 4, group B consisted of 10 patients with NYHA classification class 2, and group C consisted of 15 age-matched normal controls. Respiratory muscle strength was assessed with maximal static inspiratory mouth pressure at residual volume level and expiratory mouth pressure at total lung capacity level (PImax, PEmax, respectively). Pulmonary functions in patients with CHF showed almost normal. PImax in group A was significantly less than that in group B or C, although PImax in group B was not significantly different from that in group C. In the patients with CHF, PImax correlated positively with cardiac index and maximal oxygen consumption (r = 0.460 and r = 0.503, p < 0.05, respectively). These findings suggest that inspiratory muscle strength, which was impaired in patients with severe CHF, may be dependent on cardiac function and may be one of the limiting factors on impaired exercise capacity in the patients with CHF.

Aged↗
PubMed1994

Intravascular bronchioloalveolar tumor with skin metastases.

Intravascular bronchioloalveolar tumor (IVBAT) is a rare pulmonary tumor that occurs in the younger age groups. In the present article, we describe a patient who manifested systemic growth after the resection of the primary tumor. Skin metastasis, which has never been reported, is histologically examined.

Adult↗
PubMed1994

Derivation and application of monoclonal antibodies recognizing several epitopes on bovine serum albumin.

Three (AB-3, AB-4 and AB-6) monoclonal antibodies (mAb) to bovine serum albumin (BSA) were derived and characterized for their physicochemical and immunological properties. AB-3 recognized an epitope distinct from epitopes recognized by AB-4 and AB-6 as determined by binding inhibition assay. AB-4 and AB-6 mAbs recognized similar but not identical epitopes on BSA. Based on the antigenic specificity, we applied these mAbs to quantitative analysis of BSA in medium and to depletion of BSA from culture medium containing fetal calf serum (FCS). For quantitative analysis, we employed a sandwich enzyme-linked immunosorbent assay (ELISA) using biotinylated AB-3, solid-phase of AB-6 and an avidin-biotin-peroxidase complex system. This assay was highly sensitive and quantitative in the range of BSA concentration at 10 to 1,500 ng/ml. To deplete BSA from medium, we prepared affinity-gel coupled to AB-6. Repeated treatment of FCS-containing medium with the affinity-gel efficiently depleted BSA from the medium. The depletion capacity was 0.74 to 1.0 moles of BSA/mole of coupled mAb.

Animals↗
PubMed1994

A DNA-flow cytometric analysis of trichilemmal carcinoma, proliferating trichilemmal cyst and trichilemmal cyst.

DNA-flow cytometric analyses were performed on paraffin-embedded sections of trichilemmal carcinoma, proliferating trichilemmal cyst, and trichilemmal cyst. Nine out of 10 cases of trichilemmal carcinoma and 2 out of 5 cases of proliferating trichilemmal cyst showed aneuploidy. No aneuploidy was detected in 6 cases of benign trichilemmal cyst. The proliferating index (percentage of cells that exceeded the diploid peak (G0/G1) plus 4 standard deviations) of trichilemmal carcinoma, proliferating trichilemmal cyst, and trichilemmal cysts was 43.5 +/- 3.8%, 32.5 +/- 7.5%, and 23.1 +/- 2.0%, respectively. In one out of the 6 cases of trichilemmal carcinoma that were large enough for us to investigate regional differences, DNA ploidy heterogeneity was detected. The incidence of aneuploidy of trichilemmal carcinoma was significantly higher than that of squamous cell carcinoma (14%), previously described by us using the same method.

Adolescent↗
PubMed1994

Adjuvant activity of the cell wall of Bifidobacterium infantis for in vivo immune responses in mice.

We examined the adjuvant activity of the Bifidobacterial Cell Wall preparation (WPG) for in vivo immune responses in mice. We studied three classical immune responses, which are thought to be T-cell mediated responses, to evaluate the adjuvant activity of WPG. The delayed type hypersensitivity (DTH) responses of sheep blood red cell (SRBC)-sensitized mice were significantly augmented by WPG, although the enhancement varied with the timing, route and dosage of injection. The adjuvant activity of WPG was also confirmed by using a glutaraldehyde treated- and Concanavalin A associated- tumor vaccine (G-Con A tumor vaccine) system. BALB/c mice sensitized with G-Con A tumor vaccine and WPG improved synergistically in survival time and cure rate compared with those given G-Con A vaccine alone. Spleen cells of Meth A tumor-bearing mice induced antitumor neutralizing activity with the growth of tumor but the activity declined and disappeared at the late stage of tumor growth (over 28 days after tumor transplantation). On the other hand, antitumor neutralizing immunity was prolonged for as long as 33 days in mice inoculated with Meth A tumor and WPG. The requirement of a T-cell subpopulation in the spleen cells of tumor plus WPG treated mice was confirmed using anti-Thy 1.2 antiserum + complement to deplete them. The adjuvant activities of the Bifidobacterial cell wall demonstrated by the in vivo immune responses predict that Bifidobacteria may play a role as an immunomodulator in human and animal intestines.

Adjuvants, Immunologic↗
PubMed1994

[A case report of interstitial pneumonia caused by granulocyte colony-stimulating factor].

Several clinical trials have demonstrated that granulocyte colony-stimulating factor (G-CSF) accelerates the recovery of neutropenia in chemotherapy-induced bone marrow suppression. In this report, we describe a 46-year-old female with glioblastoma multiforme who developed interstitial pneumonia due to administration of G-CSF during the phase of immunochemoradiotherapy-induced neutropenia. Thirty-three days after starting immunochemoradiotherapy (ACNU, VCR, IFN -beta, radiation), she developed neutropenia (1,000/microliters). Administration of G-CSF at doses of 125-250 micrograms/day led to an increase of peripheral neutrophil counts. Eleven days later, the patient developed sudden severe respiratory failure and cyanosis with worsening of lung shadows. Blood gas levels on room air were PaO2 49.3mmHg, PaCO2 28.0mmHg, and pH 7.46. At this time, her neutrophil count had risen to 26,080/microliters. LDH and alpha - HBD had also increased to 1,439 IU/l and 1,117IU/l respectively. Chest radiograph and CT scan demonstrated interstitial pneumonia. After treatment with methyl prednisolone, her respiratory symptoms were gradually resolved. A number of side-effects have been reported with granulocyte-macrophage colony-stimulating factor (GM-CSF). These include fluid retention with pericardial and pleural effusion, fever, bone pain, fatigue, and rash. This report also suggests that G-CSF might be a cause of interstitial pneumonia during the phase of immunochemoradiotherapy-induced neutropenia.

Brain Neoplasms↗
PubMed1994

Stimulatory guanine nucleotide binding protein in pig epidermis: transient increase of the 45KDA cholera toxin substrate (Gs alpha) in the tape stripping-induced hyperproliferative state.

Cholera toxin catalyzed the transfer of ADP-ribose from [alpha-32P] NAD to 45kDa protein in pig epidermis. Western blot analysis using anti-Gs alpha antibody identified the 45kDa protein to be Gs alpha. In contrast to pertussis toxin-catalyzed ADP-ribosylation of Gi alpha, the cholera toxin-catalyzed ADP-ribosylation was enhanced by the presence of Mg2+ in the reaction mixture. The cholera toxin-catalyzed ADP-ribosylation of the epidermal 45kDa membrane protein was significantly decreased, when samples were prepared from the cholera toxin-pretreated epidermis. The results, coupled with our previous report (Tsutsui and Iizuka 1990), indicate that pig epidermis contains functional G proteins (Gs and Gi), that affect the epidermal adenylate cyclase activity. Tape stripping-induced hyperproliferative epidermis showed an increased cholera toxin-catalyzed ADP-ribosylation of the 45kDa protein (Gs alpha) at 12-24 h following the tape stripping. Immunoblot analysis, however, showed no remarkable change in the level of Gs alpha compared with non-stripping controls. There was no significant difference in the level of the pertussis toxin-induced ADP-ribosylation of 40kDa protein (Gi alpha) in the tape-stripped epidermis. Immunoblot analysis showed no change in Gi content, either. Forskolin-induced cyclic AMP accumulation was markedly increased in the tape stripping-induced hyperproliferative epidermis. Cholera toxin-induced cyclic AMP accumulation was slightly increased, but this was not statistically significant. These results indicate that the alteration of Gs that is documented by cholera toxin-catalyzed ADP-ribosylation, is among the functional derangements of adenylate cyclase of tape stripping-induced hyperproliferative epidermis.

Adenosine Diphosphate Ribose↗
PubMed1994

[A comparison between neuromuscular blocking effects of pipecuronium and vecuronium; a double blind controlled study in collaboration with 5 departments of anesthesiology].

Neuromuscular blocking effects of pipecuronium bromide 0.04 mg.kg-1 were compared those of vecuronium bromide 0.08 mg.kg-1 in a double blind study. Fifty surgical patients (ASA I or II) were allocated randomly to two groups of each 25 cases and they were anesthetized with nitrous oxide 4 l.min-1, oxygen 2 l.min-1 and isoflurane 1 MAC. Neuromuscular blockade was monitored by using mechanical twitch responses of the thumb to electrical stimulations of the ulnar nerve. Adequate neuromuscular relaxation for surgery of 85.8% to 100% block was obtained by this dose of pipecuronium. The duration of action and recovery time from 75% to 25% block were longer than those produced by twice the dosage of vecuronium (62.3 +/- 37.15 vs. 40.4 +/- 16.09 min, and 48.1 +/- 22.0 vs. 19.9 +/- 10.8 min, P < 0.05). The blocks by both drugs responded to neostigmine. Cardiovascular side effects of the both agents were not found. From these results, it is concluded that pipecuronium is a useful nondepolarizing relaxant with a long duration of action and negligible side effects.

Adult↗
PubMed1994

[Intrabullar suction-adhesion in a case of giant bulla of the lung with steroid--department intractable bronchial asthma].

We successfully reduced a giant bulla using an intrabullar suction-adhesion method in a case of steroid-dependent intractable bronchial asthma. A 53-year-old woman was hospitalized with an asthma attack, and suffered pneumonia in the area surrounding a giant bulla after remission of two episodes of asthma. After the pneumonia, we found that the giant bulla had increased in volume. It was readily apparent that the bulla should be reduced. At that time a right side pneumothorax occurred by accident, which improved the chest drainage. Immediately after that, we inserted a chest drainage tube directly into the bulla and instilled fibrin glue through the tube into the bulla. Intrabullar suction and bronchial embolization via bronchofiberscopy has been used to patients with inoperable giant bullae of the lung. If intrabullar suction with fibrin glue alone can reduce giant bulla, it would offer another option to patients with inoperable giant bullae without the risk of a surgical procedure.

Asthma↗
PubMed1994

[An evaluation of the hemodynamics in metastatic liver tumor by using ultrasound color Doppler flow mapping examination].

Eighty-one patients with metastatic liver tumors were studied by ultrasound color Doppler flow mapping examination. Primary origins consisted of gastric (19 cases), colorectal (40 cases), breast (7 cases), pancreatic (3 cases), gallbladder (3 cases) cancers and the others (9 cases). Metastatic lesions originated from breast, gastric and colorectal cancers had higher detection rates, however no blood flow could be observed within metastatic liver cancers from pancreatic and gallbladder cancers. There were no differences among the maximum blood flow velocity (Vmax), the resistance index (RI) and the pulsatile index (PI) in various metastatic liver tumors. In case of colorectal cancers, metastatic lesions originated from moderately differentiated adenocarcinoma had significantly lower RI (p < 0.05) and PI (p < 0.01) than that from well differentiated adenocarcinoma. In conclusion, ultrasound color Doppler flow mapping examination is a useful method for evaluation of the blood flow within metastatic liver tumors and could offer the histological differentiation of the primary origins, in case of metastatic liver cancers originated from colorectal cancers.

Adult↗
PubMed1994

[A case of emergency carotid endarterectomy to cope with vertebrobasilar insufficiency].

This is a report of an emergency carotid endarterectomy preformed on a 59 year-old male. He had been suffering from TIA of vertebrobasilar insufficiency since 1 day prior to admission. Angiography revealed severe stenosis and ulceration of the left cervical carotid bifurcation and occlusion of the bilateral vertebral arteries. Six hours after angiography, he showed hemiparesis, decerebrated posture and consciousness disturbance. These neurological signs were considered to be derived from occlusion of the left carotid bifurcation and disappearance of the collateral circulation via the left posterior communicating artery. Emergency carotid endarterectomy was performed to prevent brain infarction of the anterior and posterior circulation. Postoperative angiography showed disappearance of the stenosis of the carotid bifurcation and the recanalization of the left vertebral artery, and of the atherosclerotic stenosis of the left intracranial vertebral artery. It was considered that the left vertebral artery had been occluded by growth of a local thrombosis at the left intracranial vertebral artery.

Arterial Occlusive Diseases↗