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Biomedical subjects

H Takemura

Publications and source records attributed to H Takemura.

At least 37 records · Page 2Linked to original sources

Quantitative determination of gelatinase activity among enterococci.

Gelatinase, hemolysin and aggregation substance have all been reported to be virulence factors of enterococci. In this study, gelatinase production was investigated in isolates of Enterococcus faecalis (n=93), E. faecium (n=49) and E. avium (n=36) recovered from hospitalized patients. Gelatinase was detected in 45% of E. faecalis isolates, but could not be detected in E. faecium and E. avium. Gelatinase activity was then measured by radial diffusion for the 42 gelatinase-positive E. faecalis isolates. To convert gelatinase activity into proteinase K activity, a standard curve was produced by placing different concentrations of proteinase K into wells in the gelatine plate. Gelatinase activity per E. faecalis colony ranged from 2.6 x 10(-7) to 2.2 x 10(-5) microg/ml, proportionate to the activity of proteinase K. An approximately 84-fold difference in gelatinase concentration was observed between the colony producing the highest amount and that producing the lowest amount. This method may be useful for determining the virulence of given isolates in relation to gelatinase production as it is quick, easy and inexpensive to perform.

Adult↗

Relatedness between the coagulase gene 3'-end region and coagulase serotypes among Staphylococcus aureus strains.

The 3'-end region of the coagulase gene from 22 strains of Staphylococcus aureus including 10 standard serotype strains was sequenced, and five subgroups with 4-8 tandem repeating units were distinguished among the tested strains. Phylogenetic analysis of the 3'-end region of the coagulase gene indicated that strains belonging to the same serotype were clustered in the same branch. A phylogenetic tree of the deduced amino acid sequences revealed that the C-terminal region might not be responsible for the epitope of the coagulase protein.

Bacteriological Techniques↗

PCR-based method for isolation and detection of Chlamydia pneumoniae DNA in cerebrospinal fluids.

Since current studies indicate the possible involvement of Chlamydia pneumoniae in the pathogenesis of multiple sclerosis (MS), demonstration of C. pneumoniae in the cerebrospinal fluid (CSF) of patients with MS is highly desirable. However, there is controversy concerning the detection of C. pneumoniae in CSFs from MS patients due to the lack of a standard protocol for extraction and detection of C. pneumoniae DNA. In this regard, we attempted to establish a highly effective extraction protocol for C. pneumoniae DNA from CSFs utilizing a commercial kit and a PCR detection method. The extraction and PCR detection protocol established in this study succeeded in detecting as few as 20 C. pneumoniae organisms in 200 microl of mock CSF. The use of this protocol to detect C. pneumoniae DNA in CSFs revealed that 68% of CSF samples obtained from patients with MS were positive (11 out of 16 samples) for chlamydia DNA. Thus, the protocol established here is sensitive enough to detect chlamydia DNA from CSFs and can be used by other laboratories for evaluation of the presence of chlamydiae in CSFs because the protocol is based on the use of a commercial kit.

Chlamydophila Infections↗

[Clinical study on renal trauma].

A total of 64 patients with renal trauma were treated at the Department of Urology, St. Marianna University School of Medicine, from January 1988 to December 1996. Patients were between 3 and 66 years old (mean 28.2); 56 (87.5%) of them were male and 8 (12.5%) were female. The main cause of injury was a traffic accident in 33 cases (51.6%). According to the Classification of Renal Injury by the Japanese Association for the Surgery of Trauma in Japan, there were 27 cases (42.2%) of Type I (subcapsular injury), 12 cases (18.8%) of Type II (superficial injury), 16 cases (25.0%) of Type III (Deep injury), 6 cases (9.4%) of Type IV (Pedicle injury). Surgical treatment was performed in 6 cases (9.3%). The major associated injuries were bone fracture in 33 cases (51.6%), lung injury in 20 cases (31.3%) and liver injury in 16 cases (25.0%). The literature is also reviewed briefly.

Accidents, Traffic↗

The C-F...cation interaction: an ammonium complex of a hexafluoro macrocyclic cage compound.

An ammonium complex of the hexafluoro cage compound 1 was isolated and its structure was elucidated by X-ray crystallographic analysis. The C-F bonds are elongated by the complexation, which is clear evidence of C-F...cation interaction. The driving force of NH4(+) inclusion is the C-F cation interaction, but the C-F...HN+ hydrogen bond does not contribute to this complexation. The crystal structure of the NH4+ complex 1 shows short C-F...HN+ contacts (2.286-2.662 A). Furthermore, it shows that closer F...H(+)(-N) distances give a larger F...H(+)-N angle. Although such structural features seem to indicate the existence of C-F...HN+ hydrogen bonds, the spectral data (1H NMR, 19F VT-NMR, and IR spectroscopy) did not support the existence of hydrogen bonds. Thermodynamic parameters, log K(s) (4.6 +/- 0.1, 298 K), deltaH (-5.3 +/- 0.1 kcal mol(-1)), and deltaS (3.2 +/- 0.3 cal mol(-1) K(-1)), of the complexation were obtained in CDCl3/CD3CN mixture.

Cations↗

Synthesis of macrocyclic cage compounds by diamine-dihalide one-step coupling reaction

Macropolycyclic cage compounds were synthesized by a direct reaction between diamines and bis(bromomethyl) compounds. The procedure for constructing the polycyclic cage structure is simple and straightforward. The macropolycyclic compounds obtainable from this cyclization procedure are three-dimensional cage compounds, and any other isomers were not obtained except for two examples. Benzene, pyridine, and aliphatic units could be introduced into the cage structure. The macrocycles that have strong cation affinity were obtained as their potassium complexes.

Journal Article↗

Light chain determines the binding property of human anti-dsDNA IgG autoantibodies.

We have previously prepared human anti-double-stranded (ds) DNA IgG Fab clones using phage-display technology. Nucleotide sequence analysis of genes of immunoglobulin (Ig) heavy and light chain variable regions in these Fab clones suggested that the DNA-binding activity of the clones depended on light chain usage. To confirm the role of the light chain in antibody binding to DNA, we constructed in the present study's new recombined Fab clones by heavy and light chain shuffling between the original anti-dsDNA Fab clones. Clones constructed by pairing Fdgamma fragments with the light chain from a high DNA-binding clone showed high DNA-binding activities, whereas other constructed clones using light chains from low DNA-binding clones showed low DNA-binding activities. Our results indicate that light chains in anti-dsDNA antibodies can determine the DNA-binding activity of the antibodies. Ig chain shuffling of phage-display antibodies may be useful for investigating the molecular mechanisms for antigen-antibody binding of human autoantibodies.

Amino Acid Sequence↗

Evaluation of the activity of antimicrobial agents against Legionella pneumophila multiplying in a human monocytic cell line, THP-1, and an alveolar epithelial cell line, A549.

The intracellular activity of three antimicrobial agents, erythromycin, clarithromycin, and ciprofloxacin, against Legionella pneumophila was examined in the human monocyte-derived cell line, THP-1, and the human alveolar epithelial cell line, A549. L. pneumophila multiplied by three- to four-log in THP-1 and by two- to three-log in A549 after 48-h incubation. The activity of the two macrolides was markedly greater in A549 than in THP-1, while ciprofloxacin exhibited similar activity in both cell lines. The intracellular concentrations of the two macrolides were markedly higher in A549 than in THP-1, while those of ciprofloxacin were almost equal in both types of cell lines. The intracellular activity of antimicrobial agents and the intracellular growth of L. pneumophila vary with different types of host cells.

Anti-Bacterial Agents↗

Comparative uptake of grepafloxacin and ciprofloxacin by a human monocytic cell line, THP-1.

The present study was designed to compare the uptake of grepafloxacin by a human monocytic cell line, THP-1, with that of ciprofloxacin. THP-1 cells were incubated with 20 microg/ml of either drug, and the entry of the drugs into the cells was determined using a velocity gradient centrifugation technique and HPLC assay. Antibiotic uptake by the cells was expressed as the ratio of the intracellular to the extracellular drug concentration (IC/EC). Grepafloxacin entered THP-1 cells readily within 5 min, and at steady-state (37 degrees C; 60 min), the IC/EC ratio of grepafloxacin (11.9 +/- 1.7; n = 13) was about 2.4-fold higher than that of ciprofloxacin (5.0 +/- 1.3; n = 13). The ratios decreased at low incubation temperature (4 degrees C), in paraformaldehyde-treated dead cells, and at low extracellular pH (pH 6.0), but were not influenced by high extracellular pH (pH, 9.0). Characterization of fluoroquinolone uptake suggests that these drugs penetrate the THP-1 membrane by passive diffusion, and also, in part, via an active transport system. We also examined the uptake of the two fluoroquinolones in phorbol 12 myristate 13-acetate (PMA)-stimulated adherent THP-1 cells (THP-1 macrophages). The IC/EC ratios for both fluoroquinolones in the THP-1 macrophages were significantly higher than those in the THP-1 monocytes. Further the uptake of three other fluoroquinolones, levofloxacin, tosufloxacin, and sparfloxacin, by THP-1 monocytes was examined in comparative studies. The IC/EC ratio of grepafloxacin was comparable to that of sparfloxacin and significantly higher than that of the other fluoroquinolones. Our results indicate that grepafloxacin exhibits better intracellular accumulation than ciprofloxacin and other fluoroquinolones in human monocytic and macrophage-like cells.

Anti-Infective Agents↗

Therapeutic angiogenesis with intramyocardial administration of basic fibroblast growth factor.

BACKGROUND: Basic fibroblast growth factor (bFGF) induces endothelial cell and smooth muscle cell proliferation and stimulates angiogenesis. This study was designed to evaluate the effects of intramyocardial administration of bFGF on myocardial blood flow, angiogenesis, and ventricular function in a canine acute infarction model. METHODS: Myocardial infarction was induced in 12 dogs by ligation of the left anterior descending coronary artery. Within 5 minutes after coronary occlusion, 100 microg of human recombinant bFGF in 1 mL of saline was injected into the infarct and border zone in 6 dogs, whereas saline alone was used in 6 control dogs. Myocardial blood flow was determined with colored microspheres before and immediately after coronary ligation and again 3, 7, 14, and 28 days after treatment and it was expressed as percent of normal. Angiogenesis was evaluated by immunohistochemical studies 28 days later. Cardiac function was evaluated by repeated echocardiographic measurement. RESULTS: Treatment with bFGF significantly increased the endocardial blood flow in the border zone (7 days after infarction, 75%+/-7% and 41% +/-7% in the bFGF and control groups, respectively, p<0.01) as well as epicardial blood flow in the infarcted zone. Treatment with bFGF significantly increased the capillary density (39.7+/-2.3 and 22.7+/-1.1 vessels per visual field in the bFGF and control groups, respectively, p<0.01) as well as arteriolar density in the border zone. Treatment with bFGF significantly reduced the change in ratio of thickness of the infarcted wall to the normal wall (44%+/-6% and 26% +/-5% in the bFGF and control groups, respectively, p<0.05). It improved the left ventricular ejection fraction (7 days after infarction, 0.54+/-0.02 and 0.37+/-0.03 in the bFGF and control groups, respectively, p<0.01). CONCLUSIONS: Intramyocardial administration of bFGF increased the regional myocardial blood flow, reduced thinning of the infarcted region, and improved ventricular function in acute myocardial infarction. Intramyocardial administration of bFGF may be a new therapeutic approach for patients with acute myocardial infarction.

Animals↗

Effect of PSC 833 on the cytotoxicity and pharmacodynamics of mitoxantrone in multidrug-resistant K562 cells.

We examined the effect of PSC 833, a nonimmunosuppressive cyclosporin analogue, on the cytotoxicity, accumulation and retention of an anthraquinone antileukemia drug mitoxantrone (MIT). This was done in P-glycoprotein (PGP)-overexpressing multidrug-resistant K562/D1-9 cells and compared with the effect of cyclosporin A (CsA). We also compared MIT with the effect of PSC 833 on the cytotoxicity of daunorubicin (DNR) and doxorubicin (DOX). While PSC 833 and CsA had no effect on the cytotoxicity, accumulation and retention of MIT in the parent K562 cells, PSC 833 and CsA restored accumulation and retention of MIT in K562/D1-9 cells dose-dependently. Consequently, there was increased sensitivity of K562/D1-9 cells to MIT. The reversing activity of PSC 833 on the cytotoxicity of MIT was stronger than that of CsA, and was almost the same as the reversing activity of PSC 833 on the cytotoxicity of DNR and DOX. The resistance index of MIT decreased from 43.9-fold to 2.8-fold by 0.4 microM PSC 833, which is a clinically achievable plasma concentration. These results suggest that the combination of PSC 833 with MIT could be a promising treatment in reversing PGP-mediated MDR in leukemia patients.

Antineoplastic Agents↗

Evaluation of a human monocytic cell line THP-1 model for assay of the intracellular activities of antimicrobial agents against Legionella pneumophila.

We examined the intracellular activities of 11 antimicrobial agents against Legionella pneumophila using a human monocyte-derived cell line, THP-1. Colony counting and microscopic examination of L. pneumophila co-incubated with THP-1 cells (5 x 105 cells/well) were performed. Both extra- and intra-cellular multiplication of L. pneumophila were observed and were dependent on the inoculum of L. pneumophila in the culture; L. pneumophila did not grow in the cell culture medium alone. Light microscopic examination confirmed that extracellular L. pneumophila originated from THP-1 cells disrupted by bacterial multiplication. L. pneumophila multiplied by 3-4 logs after 24 h incubation with THP-1 cells and their number remained stable at 106-107 cfu/mL until 72 h. The results of viability studies using four antimicrobial agents-ciprofloxacin, erythromycin, minocycline and rifampicin-demonstrated that our system was suitable for the intracellular activity assay. We used a concept of 'minimum extracellular concentration inhibiting intracellular multiplication' (MIEC) to evaluate the intracellular activity of antimicrobial agents. The MIECs of three beta-lactams were markedly higher than their conventional MICs while those of macrolides, quinolones, rifampicin and minocycline were similar to their MICs. Our results suggest that evaluation of the clinical efficacy of drugs against L. pneumophila should include determination of their intracellular activity against the bacteria, which could be measured using our assay system in THP-1 cells.

Anti-Bacterial Agents↗

Palmar fasciitis and polyarthritis associated with gastric carcinoma: complete resolution after total gastrectomy.

Palmar fasciitis and polyarthritis (PFA) is a rare paraneoplastic rheumatic syndrome characterized by flexion contractures of both hands and thickening of palmar fascia. Several reports have suggested that this syndrome is a tumor-associated autoimmune disorder. We report a 44-year-old Japanese man who presented with flexion contractures of both hands associated with thickening of palmar fascia and polyarthritis. These clinical pictures were suggestive of PFA associated with occult neoplasm. Upper gastrointestinal endoscopic examination revealed advanced gastric cancer. Resection of the cancer resulted in a gradual resolution of palmar fasciitis and polyarthritis. This clinical course suggests an underlying tumor-related immunologic process in this syndrome.

Adenocarcinoma↗

[A case report of Binswanger type encephalopathy manifested after coronary artery bypass grafting].

A 68-year-old man showed signs of dementia after undergoing triple CABG with cardiopulmonary bypass. Brain CT revealed widespread atrophy and lucency in white matter without any sign of stroke. No critical stenosis in cervical vessels was found in duplex scanning. He kept politeness well, but had severe memory impairment. These findings were characteristic of Binswanger type encephalopathy which must have been latent preoperatively. This disease is caused by ischemic damage in brain stem and white matter due mainly to atherosclerotic micro cerebrovasculopathy, and highly related to hypertension. We have to be aware of intracranial cerebrovascular disease when assessing the risk of brain damage in candidates for CABG.

Aged↗

The relationship between serum immunoglobulin levels and pulmonary involvement in systemic sclerosis.

OBJECTIVE: To examine the relationship between serum immunoglobulin (Ig) levels and pulmonary function in patients with systemic sclerosis (SSc). METHODS: Twenty-four patients with SSc who had at least 2 sets of pulmonary function tests (PFT) at intervals of more than one year were eligible. Multiple linear regression models were constructed for prediction of the annualized rates of change of forced vital capacity (FVC), carbon monoxide diffusing capacity (DL(CO)), and DL(CO) per unit alveolar volume (K(CO)). RESULTS: The rates of change of FVC and K(CO) correlated with the annualized rate of change of IgG (p < 0.001 and p = 0.005, respectively), and the rate of change of DL(CO) correlated with the serum IgM level at the first PFT (p = 0.020) and with the annualized rate of change of IgG (p = 0.007). CONCLUSION: The rates of change of serum Ig levels are associated with those of pulmonary function in SSc. Use of this model may assist investigation of pulmonary involvement.

Adult↗

[Clinical study of transurethral surgery for benign prostatic hyperplasia using a thick loop (VaporTome)].

From December 1996 to June 1998, we performed transurethral surgery of the prostate using a thick loop, VaporTome developed by Circon ACMI on 23 patients with symptomatic benign prostatic hyperplasia (BPH). The mean international prostate symptom score decreased significantly, from 22.6 at baseline to 9.0, 6.5, 4.9 and 5.3 at 1, 3, 6 and 12 months after operation, respectively. The mean quality of life index also decreased significantly, from 5.5 to 1.8, 1.8, 1.4 and 1.2, respectively. The mean peak flow rate increased significantly, from 6.2 preoperatively to 15.8, 17.1, 16.6 and 17.4 ml/sec at 1, 3, 6 and 12 months after operation, respectively. The mean prostate volume decreased significantly, from 66.9 to 24.8, 17.3, 19.0 and 18.2 ml. The mean residual urine decreased significantly, from 167.9 to 11.4, 9.4, 12.8 and 20.4 ml. As for overall efficacy, the rate of excellent and good cases at 1, 3, 6 and 12 months was 80.9, 89.4, 94.7 and 85.8%, respectively. No serious complications were observed. Our clinical results suggest that transurethral surgery for BPH using VaporTome has several potential advantages including high efficacy, minimal morbidity compared with standard transurethral resection of the prostate and lower cost compared with other minimally invasive procedures.

Aged↗

[A case of recurrent spontaneous vesical rupture subsequent to irradiation for uterine cancer].

The patient, a 68-year-old woman, had undergone radical hysterectomy and post-operative radiotherapy for uterine cancer in 1974. She was admitted to our hospital complaining of abdominal pain in February, 1996. Contrast-enhanced computed tomographic scan and cystography showed leakage of contrast medium around the bladder. We diagnosed her with spontaneous vesical rupture, and performed conservative therapy. Two months later, she was re-admitted with recurrent vesical rupture. We again performed conservative therapy. We consider that conservative therapy can be indicated for spontaneous vesical rupture with good general condition and no severe urinary tract infection.

Aged↗

[Outcomes and predictors of mortality in elderly patients requiring artificial ventilation].

We retrospectively examined the outcomes and the predictors of mortality in 97 patients aged 70 years and over (mean: 79.3 years) who required artificial ventilation for more than 3 hours. The median duration of artificial ventilation was 16 days (range: 1-85). Of these patients, 61% survived ventilator weaning and 37% were discharged from hospital alive. We performed univariate and logistic regression analysis to determine the predictors of dying before weaning and hospital discharge using severity of illness data. The predictors of hospital mortality were examined in 86 patients, excluding those who had malignant disease, all of whom died in hospital. Activities of daily living (ADL) were ranked as "bedridden", "in wheelchair", or "independent". In the three age groups-up to 70 years, 75 to 84 years and 85 years and over-the respective survival rates were 63% (weaned) and 67% (discharged), 69% (weaned) and 39% (discharged), and 33% (weaned) and 12% (discharged); the overall p values being 0.026 (weaned) and 0.003 (discharged). The predictors of dying before weaning according to univariate analysis were as follows: age (p = 0.026), respiratory or cardiac arrest on admission (p = 0.003), acute physiology score (APS) of 25 or more on admission (p = 0.000), systolic blood pressure below 90 mmHg on admission (p = 0.001), hemoglobin less than 11 g/dl (p = 0.044), and total protein less than 6 g/dl (p = 0.007). The predictors of hospital mortality by univariate analysis were as follows: age (p = 0.003), limited ADL (p = 0.001), respiratory or cardiac arrest on admission (p = 0.011), APS 25 or more on admission (p = 0.049), systolic blood pressure less than 90 mmHg on admission (p = 0.002), hemoglobin less than 11 g/dl (p = 0.028), and GOT or GPT more than 50 IU (p = 0.038). The relative risk of dying before weaning decreased in the order: respiratory or cardiac arrest on admission, systolic blood pressure less than 90 mmHg on admission, total protein less than 6 g/dl (Odds ratios: 6.04, 3.90, 3.51, respectively), or, respiratory or cardiac arrest on admission, APS more than 25 in admission, total protein less than 6 g/dl (Odds ratio: 6.94, 3.99, 3.76, respectively). The relative risk of hospital mortality decreased in the order: "bedridden", systolic blood pressure less than 90 mmHg on admission, "with wheel chair" (Odds ratios: 11.76, 6.44, 3.57, respectively). In the older patients, successful ventilator weaning was not indicative of hospital discharge. Ventilator weaning depended mainly on acute health status on admission, but hospital discharge depended also on the presence of limited ADL and preexisting malignant disease.

Activities of Daily Living↗