Biomedical subjects

H Masaki

Publications and source records attributed to H Masaki.

At least 163 records · Page 9Linked to original sources

PubMed1995

[A case of thymic squamous cell carcinoma successfully treated with curative resection].

A case of rare thymic squamous cell carcinoma was reported. A 46-year-old female was admitted to our hospital because of an abnormal shadow on chest X-ray. Chest CT showed anterior mediastinal tumor and histological diagnosis of squamous cell carcinoma was made by needle biopsy under CT guide. Esophagus and lung were no abnormal findings, we thought the primary region was thymus. On mid-sternotomy, anterior mediastinal tumor was resected with thymus, right phrenic nerve and pericardium, however it was not direct invasion to heart, great vessels, lung and chest wall. Resection of peripheral fatty tissue and dissection of mediastinal lymph nodes as much as possible, it was not capsular invasion and mediastinal lymph nodes metastasis and complete curative resection was able to perform. Additional radiation therapy was done, post-operative course was uneventful.

Carcinoma, Squamous Cell↗

PubMed1995

An electron microscopic study of the processes of nerve regeneration after lyophilized nerve homografting.

The objective of the present study was to investigate the role of the Schwann cell basal lamina in nerve regeneration. To achieve this goal, we observed the process of axonal regeneration within a lyophilized nerve graft, in which only the basal lamina of the Schwann cell persisted. Sciatic nerves were removed from rats and lyophilized to kill the Schwann cells and other components. These grafts were transplanted to rat sciatic nerve defects. The rats were then killed after lapses of time. We observed the processes of axonal regeneration using a transmission electron microscope. Regeneration of axons along the inner surface of the Schwann cell basal lamina was clearly seen. These results suggest that, if tubular basal laminae persist, Schwann cells are not always necessary, and axonal regeneration can be induced in the direction toward the basal lamina.

Animals↗

PubMed1995

Beta 2-adrenergic receptor regulation of the cardiac L-type Ca2+ channel coexpressed in a fibroblast cell line.

To characterize the functional coupling of the beta 2-AR to the cardiac Ca2+ channel in a system with a single receptor subtype, we stably cotransfected a Chinese hamster fibroblast (CHW) cell line, which lacks beta 2-ARs and Ca2+ channels, with the rabbit cardiac Ca2+ channel alpha 1 and beta 2 subunits and the human beta 2-AR cDNAs. The effects of beta 2-AR stimulation on the expressed Ca2+ channel current were examined using the whole-cell patch-clamp technique. CHW cells transfected with the Ca2+ channel subunits displayed a voltage-dependent inward current having properties typical of native cardiac L-type Ca2+ channels. The expressed current was increased by a phosphorylation-dependent mechanism. CHW cells cotransfected with the Ca2+ channel subunits and the beta 2-AR were responsive to isoproterenol (Iso) in a dose-dependent manner. Iso (10 microM) increased peak Ca2+ channel current to 172 +/- 5% (n = 17) of control amplitude, indicating that the expressed Ca2+ channels are functionally coupled to the beta 2-AR. The results demonstrate unequivocally that beta 2-ARs can modulate the activity of cardiac Ca2+ channels, independent of beta 1-ARs. The results also demonstrate the usefulness of the CHW heterologous expression system, the first to reconstitute physiological modulation of an L-type Ca2+ channel by the beta 2-AR, for studying receptor subtype-specific regulation of the Ca2+ channel.

Adrenergic beta-Agonists↗

PubMed1994

Purification and analysis of a flavoprotein functional as NADH oxidase from Amphibacillus xylanus overexpressed in Escherichia coli.

The gene encoding the Amphibacillus xylanus flavoprotein has been cloned into pTTQ18 and overexpressed in Escherichia coli. The recombinant enzyme has been purified to homogeneity yielding 15 mg of pure enzyme/liter of cell culture. Recombinant flavoprotein is fully active and has an absorption spectrum identical to that of the enzyme purified from A. xylanus. The N-terminal sequence analysis and analytical gel filtration data confirm the structural identity of recombinant and A. xylanus enzymes. The Km value for oxygen and the Km value for NADH are 1.7 mM and 33.3 microM, respectively. In the presence of free additional FAD, however, the Km value for oxygen decrease dramatically. The NADH oxidase activity is accelerated markedly in the presence of additional FAD. The intracellular free FAD concentration of A. xylanus is calculated about 13 microM. This FAD concentration would be enough to accelerate the NADH oxidase activity of flavoprotein in cells of A. xylanus. Two-electron reduction of the enzyme FAD by the strong reductant dithionite occurs during the total uptake of 6 electrons. Such behavior usually indicates the presence of non-flavin redox centers. The high degree of homology between this enzyme and alkyl hydroperoxide reductase F52a protein and thioredoxin reductase suggests that these centers are the redox-active disulfide adjacent to the FAD and another disulfide, which is able to slowly interchange with the redox-active disulfide. The presence of two disulfides has been demonstrated.

Amino Acid Sequence↗

PubMed1994

A model system for the continuous production of a heterologous protein using a novel secretion promoting factor which operates in Escherichia coli.

The 'PAS factor' whose gene has been cloned from a species of Vibrio, is a novel protein secretion factor which is functional in Escherichia coli cells. To demonstrate that practical use of the PAS factor gene is possible, we have constructed a model secretion vector 'pAS23'. Using this system, beta-lactamase was produced and secreted into the medium of a continuous culture system, after optimization of culture conditions.

Bacterial Proteins↗

PubMed1994

A novel protein secretion factor from a Vibrio species which operates in Escherichia coli.

A DNA fragment specific to a Vibrio species was found to promote extracellular secretion of proteins, when cloned into Escherichia coli. Cells harboring a plasmid carrying this fragment secreted significant amounts of periplasmic beta-lactamase and alkaline phosphatase into the medium, however most cytoplasmic beta-galactosidase was retained within the cell. The DNA sequence essential for this property was found to be a gene encoding 76 amino acids, which was designated as the 'PAS factor'. Highly expressed PAS factor is harmful to the cell, this may be due to a disruption of the membrane structure and/or function.

Amino Acid Sequence↗

PubMed1994

Replacement of an amino acid residue of cyclodextrin glucanotransferase of Bacillus ohbensis doubles the production of gamma-cyclodextrin.

Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) produces cyclodextrin (CD) from starch through an intramolecular transglucosylation reaction. To obtain a better understanding of the amylolytic and cyclization mechanisms of CGTase, and furthermore to improve the production of gamma-CD, mutant CGTases were constructed by site-directed mutagenesis of the CGTase gene of Bacillus ohbensis replacing Tyr at position 188 by 19 other amino acids. All mutant enzymes retained both starch-degrading and CD synthesizing activities to various extents. Among them, a mutant enzyme having Trp instead of Tyr-188 produced 15% of gamma-CD from soluble starch, which is about twice as much as the amount produced by the wild-type enzyme.

Amino Acid Sequence↗

PubMed1994

Construction of squalene-accumulating Saccharomyces cerevisiae mutants by gene disruption through homologous recombination.

Saccharomyces cerevisiae synthesizes ergosterol via squalene, but squalene is hardly detected in aerobically grown cells. To obtain a stable squalene-accumulating yeast strain, we attempted to disrupt a gene required in the conversion of squalene to ergosterol, by homologous recombination with a short piece of the gene fragment conjugated with an integration plasmid vector carrying the LEU2 gene. Two mutants that required ergosterol at least for fast growth were isolated. In an aerobic cultivation and with ergosterol supplementation, the two mutants accumulated squalene up to 5 mg/g dry cells. Southern hybridization analysis indicated that both mutants had acquired the vector DNA integrated in the same gene, or nearby genes, on chromosome 12.

Biotechnology↗

PubMed1994

Expression of fimbriae and host response in Branhamella catarrhalis respiratory infections.

Sputum during the acute exacerbation of chronic respiratory diseases were observed under the electron microscope, to determine the in vivo expression of surface structures of Branhamella catarrhalis (B. catarrhalis), the polymorphonuclear neutrophil (PMN) response to B. catarrhalis infections, and the composition of sputum. It was found that during infection fimbriae are expressed in B. catarrhalis. However, there were sparsely to densely fimbriated bacteria in each sputum sample. The length of the fimbriae were from 50 to 76 nm. In the sparsely fimbriated B. catarrhalis, external to the cell wall, a thin, granular, electron-dense layer was observed. Due to the presence of fimbriae, this layer was not seen in densely fimbriated B. catarrhalis. Blebs were also found in B. catarrhalis. PMNs were found to phagocytose both B. catarrhalis and debris. Evidence was found that debris were formed mainly by the destruction of PMNs. Bacteria as well as debris were phagocytosed by PMNs.

Adult↗

PubMed1994

[Clinical significance of positive blood cultures in the Nagasaki Rosai Hospital during the past 6 years].

In this clinical study we tried to find out the clinical significance of positive blood cultures between Jan. 1984 and Dec. 1989 in the Nagasaki Rosai Hospital. In this period the association between the isolated bacteria from blood cultures (398 specimens: 17.9%) and clinical background were evaluated. Each year the positive cultures were seen in about 18-20%. The distribution of bacteria showed increase percentage of the gram-positive organisms such as coagulase-negative Staphylococcus. Gram-negative organisms in included P. aeruginosa, A. calcoaceticus and E. coli. A. calcoaceticus was frequently found from the dept. of neurosurgery. Fungus, mostly Candida, was isolated in about 10-20%. The specimens were provided from department of internal medicine (62.9%) and neurosurgery (22.8%). An analysis of 213 episodes of 188 bacteremic cases was made. The onset of bacteremia had two peaks, 24 h or 1 month, after admission. The primary infected sites were the urinary tract (26.7%), respiratory system (14.5%) and central nervous system (6.1%). The central venous catheter was an important infected site. The major underlying diseases included acute cerebrovascular diseases, cardiac failure due to valvular heart diseases or acute myocardial infarction and/or iatrogenic immunocompromised state. The major complications were disseminated intravascular coagulation (DIC) (24.4%) and multiple organ failure (MOF) (8.9%). Half of the DIC episodes was caused by gram-positive cocci such as CNS, E. faecalis and MRSA. 68.4% of MOF episodes was associated with gram-negative bacteria. While the remainder was caused by gram-positive cocci including CNS and MRSA.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

PubMed1994

Cloning, sequencing, and expression of thermophilic Bacillus sp. strain TB-90 urease gene complex in Escherichia coli.

The urease of thermophilic Bacillus sp. strain TB-90 is composed of three subunits with molecular masses of 61, 12, and 11 kDa. By using synthetic oligonucleotide probes based on N-terminal amino acid sequences of each subunit, we cloned a 3.2-kb EcoRI fragment of TB-90 genomic DNA. Moreover, we cloned two other DNA fragments by gene walking starting from this fragment. Finally, we reconstructed in vitro a 6.2-kb DNA fragment which expressed catalytically active urease in Escherichia coli by combining these three DNA fragments. Nucleotide sequencing analysis revealed that the urease gene complex consists of nine genes, which were designed ureA, ureB, ureC, ureE, ureF, ureG, ureD, ureH, and ureI in order of arrangement. The structural genes ureA, ureB, and ureC encode the 11-, 12-, and 61-kDa subunits, respectively. The deduced amino acid sequences of UreD, UreE, UreF, and UreG, the gene products of four accessory genes, are homologous to those of the corresponding Ure proteins of Klebsiella aerogenes. UreD, UreF, and UreG were essential for expression of urease activity in E. coli and are suggested to play important roles in the maturation step of the urease in a co- and/or posttranslational manner. On the other hand, UreH and UreI exhibited no significant similarity to the known accessory proteins of other bacteria. However, UreH showed 23% amino acid identity to the Alcaligenes eutrophus HoxN protein, a high-affinity nickel transporter.

Alcaligenes↗

PubMed1994

Dynamic arterial baroreflex in rabbits with heart failure induced by rapid pacing.

Excessive sympathetic nerve activity in heart failure could be attributable to impaired arterial baroreflex function. Employing transfer function analysis, we evaluated the arterial baroreflex in control rabbits (n = 8) and in rabbits with rapid pacing-induced heart failure (n = 10) in a dynamic manner. Rabbits in the heart-failure group showed elevated filling pressures, depressed first derivative of left ventricular pressure, pulmonary congestion, and an increased level of plasma norepinephrine. Varying aortic pressure pseudorandomly and recording responses in renal nerve activity, we calculated the transfer function from aortic pressure to renal nerve activity. The gain of the transfer function was similar between control and heart-failure rabbits over 0.04-0.4 Hz as well as the phase and the coherence, indicating that the dynamic arterial baroreflex was preserved in our rabbit heart-failure model. Vagotomy increased the gain of the arterial baroreflex over 0.04-0.4 Hz in control (P < 0.05) but not in heart-failure rabbits, indicating that vagal afferents, which normally inhibit the dynamic arterial baroreflex, no more did so in heart failure. We conclude that excessive sympathetic nerve activity in heart failure may not be due to impaired dynamic arterial baroreflex, but that this apparently preserved arterial baroreflex in heart failure may be due to impaired cardiopulmonary baroreflex.

Animals↗

PubMed1994

cAMP responsive element-mediated regulation of the gene transcription of the alpha 1B adrenergic receptor by thyrotropin.

To elucidate the molecular mechanism of the stimulatory effect of thyrotropin on the gene regulation of alpha 1B adrenergic receptor in functioning rat thyroid (FRTL-5) cells, we established a competitive reverse-transcriptase (RT) polymerase chain reaction (PCR) and nuclear run-off assay to quantify changes in mRNA levels and transcription rates. A binding assay showed that FRTL-5 cells predominantly expressed alpha 1B adrenergic receptor and that thyrotropin increased its expression sevenfold. By means of RT-PCR, we found that thyrotropin induced an 11-fold increase in alpha 1B receptor mRNA abundance. The nuclear run-off assay demonstrated that thyrotropin caused a ninefold increase at the gene transcriptional level, which occurred in the presence of the protein synthesis inhibitor cycloheximide. The half-life of the alpha 1B receptor mRNA in cells incubated with thyrotropin for 1 h increased 1.5-fold but returned to the original value after 12 h. Dibutyryl cAMP and forskolin mimicked the stimulatory effects of thyrotropin on the gene transcriptional level. The 5'-flanking region of the rat alpha 1B receptor gene contained a putative cAMP responsive element (CRE) at nucleotide -438 relative to the translation start site. The promoter analysis using the reporter gene indicated that the CRE motif confers the cAMP sensitivity to the transcription of the rat alpha 1B receptor gene. These results demonstrated that a CRE-mediated mechanism is involved in the transcriptional regulation of the alpha 1B receptor gene by thyrotropin without requiring new protein synthesis.

Animals↗

PubMed1994

Influence of arotinolol hydrochloride on heart rate spectrum in hypertensive subjects.

Influence of arotinolol hydrochloride and atenolol on the balance between the sympathetic and parasympathetic nervous systems was evaluated in 8 hypertensive subjects by spectral analysis of heart rate (HR) and systemic blood pressure (BP). Before and after administration of either arotinolol (n = 7) or atenolol (n = 7) for 2 weeks, BP was continuously and non-invasively monitored by a finger-cuff manometry (Finapres). A time series of instantaneous HR was constructed from the BP signal. A time series of mean BP was also constructed. Spectral analysis was performed by the use of an autoregressive algorithm on these time series (approximately 180 sec). Each spectrum was subdivided into low-(0.05-0.15 Hz, LF) and high-frequency (0.15-0.4 Hz, HF) components, and each component was divided by the sum of the two for normalization. As a measure of the balance between the sympathetic and parasympathetic nervous systems, the ratio of LF to HF (LF/HF) was evaluated. Arotinolol increased fractional HF in the HR spectrum from 0.45 +/- 0.12 to 0.73 +/- 0.08 (p < 0.01) and decreased fractional LF from 0.55 +/- 0.12 to 0.27 +/- 0.08 (p < 0.01); consequently, it decreased LF/HF from 1.4 +/- 0.5 to 0.4 +/- 0.2 (p < 0.01). Atenolol had similar effects on these parameters. Neither of these beta-adrenergic blockades produced a discernible decrease in LF/HF in the BP spectrum. In conclusion, these beta-adrenergic blockades decreased LF/HF in the HR spectrum in hypertensive subjects, which suggests that they improved the balance between the sympathetic and parasympathetic nervous systems.

Adrenergic beta-Antagonists↗

PubMed1994

Cloning of a new cryIA(a) gene from Bacillus thuringiensis strain FU-2-7 and analysis of chimaeric CryIA(a) proteins for toxicity.

We cloned the cryIA(a) gene from Bacillus thuringiensis strain FU-2-7, one of the toxin genes encoding lepidopteran-specific protoxins. Sequence analysis of the gene showed two amino acid differences (Pro77 to Leu and Phe965 to Ser) from the CryIA(a) of B. thuringiensis strain HD-1. We constructed chimaeric cryIA(a) genes using FU-2-7 and HD-1 cryIA(a) genes and isolated the chimaeric protoxins, as well as the parental ones, from Escherichia coli cells harboring the recombinant plasmids to examine the effects of the two amino acid variations on the toxicity. FU-2-7 CryIA(a) protein was about half as toxic against the smaller tea tortrix, Adoxophyes sp., and one-third as toxic against the silkworm, Bombyx mori, as that of HD-1. On the other hand, a chimaeric CryIA(a) protein with a single replacement of Phe965 to Ser had nearly the same toxicity as the HD-1 CryIA(a) against the smaller tea tortrix and one-third the toxicity against silkworm as that of HD-1. This improved property of the chimaeric CryIA(a) protoxin may be useful for widening its application to crop protection in sericultural countries.

Amino Acid Sequence↗

PubMed1994

[Reoperation after open mitral commissurotomy for mitral stenosis].

Between 1975 and 1993, 16 of 95 patients who received open mitral commissurotomy for mitral stenosis required reoperation for recurrent mitral lesions with a mean duration of 11 years after the initial operation at Kawasaki Medical School Hospital. The mitral lesions necessitating reoperation involved restenosis in eight, stenoinsufficiency in six and regurgitation in two. In 13 patients, mitral commissure was well separated, and the mitral restenosis and regurgitation were caused by progressions of valvular and subvalvular lesions. Significant tricuspid valve regurgitation was also seen in nine patients, and in seven out of eight patients who were in NYHA functional class III or IV, tricuspid regurgitation of grade 3 was observed. The combined tricuspid regurgitation aggravated the patient's symptoms and became a major risk factor of the reoperation after open mitral commissurotomy.

Catheterization↗

PubMed1994

[The structural paradigm of nursing care for patients with chronic illness: clinical experience with diabetic outpatients (III)].

The purpose of this study was to clarify a nursing paradigm of care for patients with chronic illness. Data were derived from fieldnotes of direct patient care for 111 diabetic outpatients. These fieldnotes included nursing intervention, conversation process, patient response to the nurse, the nurse's decision making process, and the nurse's reflections after intervention. Interpretive analysis identified nursing goals, nursing interventions, clinical judgment, and the nature of the patient-nurse relationship. Five nursing goals of patient-oriented care were identified: compliance, self-management, self-acceptance, independence, and self-actualization. Five approaches for nursing intervention also were identified: directed, facilitating, acknowledging, mutually responding, and mutually cooperating. From these findings, a nursing paradigm of care was conceptualized. As nursing goals became more "holistic", the patient-nurse relationship became more intimate, and the patients' competency to participate in the management of their own diabetic care became greater. This paradigm should assist nurses in judging and selecting the proper care for the needs of each patient.

Chronic Disease↗

PubMed1994

Detection of minimal residual disease using clonospecific primers for CDRIII in patients with acute B lymphocytic leukemia with or without Philadelphia chromosome: possibility of clinical application as a tool for improving prognosis.

We attempted to identify the minimal residual leukemic clone as related to the clinical course in patients with acute B lymphocytic leukemia (B-ALL). DNA was extracted from stored bone marrow slides, and the third complementarity determining region (CDRIII) was amplified by polymerase chain reaction (PCR) using primers with consensus sequences for VH and JH. After amplification of the CDRIII band, the DNA fragment of CDRIII was inserted into the cloning vector PUC118. After cloning, the DNA sequences for CDRIII were determined. Clonospecific DNA sequences in CDRIII were selected, and clonospecific primers for each patient were synthesized. Using the clonospecific primers, we carried out second-round PCR to detect minimal residual disease (MRD) during several stages of the clinical course. Basically, the sensitivity of detection for MRD was between 10(-4) and 10(-5) cells. Even when leukemic cells were not detected in the morphologic study, with this detection system, the MRD was identified as an amplified CDRIII band stained with ethidium bromide on agarose gel. After bone marrow transplantation (BMT), MRD was detected for at least 4 months. In this article, we discuss the difference in sensitivity of detection for MRD between the BCR-ABL fusion gene and CDRIII in Philadelphia chromosome-positive (Ph+) B-ALL, as well as the possible clinical application of this method to predict relapse and prognosis.

Adolescent↗