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Biomedical subjects

H Kobayashi

Publications and source records attributed to H Kobayashi.

At least 253 records · Page 14Linked to original sources

Induction of unscheduled DNA synthesis in hairless mouse epidermis by 8-methoxypsoralen plus ultraviolet A (PUVA).

Induction of unscheduled DNA synthesis (UDS) by 8-methoxypsoralen (8-MOP) plus ultraviolet A (UV-A) (PUVA) was investigated in the epidermis of female hairless mice by means of an in vivo--in vitro assay using a liquid scintillation counting method. Groups of three to five 8-week-old female hairless mice had 8-MOP applied once onto two areas of the back after stripping of the stratum corneum with adhesive tape to enhance skin penetration, and were irradiated with UV-A. Skin samples were taken and cultured in a medium containing [3H]thymidine with or without hydroxyurea (HU) for 2 hr. DNA of the epidermis was extracted, and the incorporation of [3H]thymidine into DNA and the DNA content were determined with a liquid scintillation counter and a fluorescence spectrophotometer, respectively. Induction of UDS was judged in terms of the UDS index [(the ratio of DNA synthesis in the presence of HU to that in its absence) x 100]. In a time-course study, the UDS index was increased at 1, 2 and 24 hr after 1 x 10(5) J/m2 UV-A irradiation with 0.001% 8-MOP, reaching the maximum level at 24 hr. In a dose-response study, it was significantly increased at the dose of 1 x 10(5) J/m2 of UV-A at 24 hr with 0.001% 8-MOP, but showed no significant change at the doses of 0.5 x 10(5), 2 x 10(5) and 4 x 10(5) J/m2. In a further study on the effect of varying the dose of 8-MOP, the UDS index was significantly increased at 0.001 and 0.002% 8-MOP at 24 hr after 1 x 10(5) J/m2 UV-A irradiation, reaching the maximum level with 0.002% 8-MOP. The increase of the UDS index in these studies was less than 3-fold. These results show that PUVA causes a small induction of UDS, which might be due to slow DNA excision repair over a long period.

Animals↗

Malignant lymphoma of the bone associated with systemic sarcoidosis.

A 57-year-old woman was hospitalized with malignant lymphoma of the right talus. After treatment, complete remission was obtained. Gallium-67 scintigraphy to confirm the remission demonstrated increased uptake in the whole body skeletal muscle, especially in her thighs. Biopsy of right gastrocnemius muscle showed epithelioid granuloma. Serum angiotensin-converting enzyme activity (ACE) and lysozyme had increased to several times the normal range. We diagnosed her disease as bone-associated sarcoidosis-lymphoma syndrome. Human herpes virus 8 (HHV-8) genome was examined in the bone marrow specimen, and the relationship between sarcoidosis-lymphoma syndrome and HHV-8 was discussed.

Antineoplastic Combined Chemotherapy Protocols↗

Primary racemose hemangioma of the bronchial artery.

A 44-year-old woman was admitted to our hospital for repeated hemoptysis. Selective angiography of the bilateral bronchial arteries showed them to be enlarged and convoluted, and revealed shunts between the bronchial and pulmonary arteries. The patient was diagnosed as having racemose hemangioma of the bronchial artery. Although embolization of the bronchial artery with gelatin sponges was performed, hemoptysis emerged again about one year later. Finally, segmentectomy of the right lower lung was performed and primary racemose hemangioma of the bronchial artery was histopathologically confirmed.

Adult↗

IgA-kappa type multiple myeloma affecting proximal and distal renal tubules.

A 45-year-old male was admitted because of chest pain, lumbago, and bilateral ankle pain. Examination disclosed hypophosphatemic osteomalacia, acquired Fanconi syndrome, and abnormalities in distal nephron such as distal renal tubular acidosis and renal diabetes insipidus. Further exploration revealed IgA kappa multiple myeloma excreting urinary Bence Jones protein (kappa-light chain). Renal biopsy revealed thick basement membranes and elec-tron-dense crystals in proximal tubular epithelial cells. Immunofluorescent studies revealed deposition of kappa-light chain in renal tubular epithelial cells that caused the renal tubular damage. Although the osteomalacia was relieved by medical treatment, the urinary Bence Jones protein and the renal tubular defects were not improved by the chemotherapy for the myeloma. The patient died of exacerbation of multiple myeloma at 50 years of age.

Biopsy↗

Lupus anticoagulant in myasthenia gravis associated with IgM gammopathy.

The plasma of a patient with myasthenia gravis had strong lupus anticoagulant activity and his IgM paraprotein displayed non-specific inhibition to coagulation factors IX, XI, XII, prekallikrein, and high molecular weight kininogen. He was placed on prednisolone, which resulted in improvement in his myasthenic symptoms, but the prolongation of APTT and macroglobulinemia remained. Double filtration plasmapheresis successfully decreased the serum IgM level from 1,190 mg/dl to 375 mg/dl and APTT improved from 58 s to 38 s. Myasthenia gravis is frequently associated with other autoimmune diseases, but the association with lupus anticoagulant and IgM gammopathy is rare.

Humans↗

Acute changes in bronchoconstriction influences exhaled nitric oxide level.

In previous studies the exhaled nitric oxide (NO) level of asthma patients was investigated only in association with bronchial inflammation, and whether the degree of bronchoconstriction itself influences the exhaled NO level has never been investigated. We therefore evaluated the effect of inhalation of a bronchoconstrictor (methacholine) or a bronchodilator (salbutamol) on the exhaled NO level of healthy volunteers and asthma patients. The exhaled NO level of the healthy volunteers decreased after methacholine inhalation. The exhaled NO level of patients with mild or moderate persistent asthma, who had no asthma attacks on the day of measurement, increased after salbutamol inhalation, and the exhaled NO level of asthma patients during asthma attacks increased after salbutamol inhalation followed by intravenous drip infusion of aminophylline. It is suspected that large amounts of NO are trapped in the lung distal to the constricted airway, contributing little to the exhaled NO level at the mouth. However, we expect that the trapped NO is exhaled at a larger fraction after the dilatation of the constricted small airway, thereby increasing the exhaled NO level at the mouth. In conclusion, the results of this study suggest that acute changes in bronchoconstriction themselves influence the exhaled NO level independently of the change in NO synthase activity associated with airway inflammation.

Adult↗

Separation of DNA by free flow electrophoresis in space.

Free flow electrophoresis of a nematode C. elegans DNA was carried out on the space shuttle flight in International Microgravity Laboratory No. 2 (IML-2)., 1994. We selected the C. elegans DNA as the sample of the space experiment for free flow electrophoresis separation. This worm is a useful animal for the study of development and behavior by genetic analysis, and is a good candidate for a complete DNA sequence analysis because the haploid genome size consists of approximately 100 Mb (megabase) distributed on the six chromosomes, only 1/30 of human genome size. (Recently, the entire genomic DNA (97 Mb) sequences of C. elegans have been completed at the last December, 1998, as the first organism of multicellular system. In addition, many of the genes in C. elegans have extensive similarity to their mammalian counterparts. It may be possible to use the technology of free flow electrophoresis to contribute to the DNA analysis of the eukaryote. In this mission we attempted to make real-time communication between the space shuttle and on the ground during the electrophoresis processing. Because the separation tendency of the sample in space was not predicted so perfectly, the requests to the crews of the space shuttle was needed for selecting the fractions to be separated. According to the three dimensional electropherogram (3DEP) figured out the electrophoresis behavior, we were able to recover those fractions and kept in the deep freezer until landing. This real-time monitoring of the electrophoresis was the first evidence during space electrophoresis experiments which was started at Apollo 14, 1974. Unfortunately, some troubles occurred by contamination of bubbles in space nevertheless the post flight analyses of the fractionated samples were succeeded. The DNAs estimated by the DNA probes were not isolated one but the migration tendencies were differ from each other. It was clear that certain parts of the process in this study are advanced; that is, the precise monitoring of the electrophoresis process in space was accomplished, in real-time, by using 3DEP. In addition, the detection of DNA of recovered samples indicates that all the processes during the study were kept to be free from biological contamination. These results were published in following paper and reviewed in the book.

Animals↗

Neuroprotectins A and B, bicyclohexapeptides protecting chick telencephalic neuronal cells from excitotoxicity. I. Fermentation, isolation, physico-chemical properties and biological activity.

Glutamate, an excitatory amino acid, is known to induce neurotoxicity in central nervous system under abnormal conditions such as ischemia, hypoglycemia, epilepsy, Huntington's chorea, Parkinson's disease and Alzheimer's disease. In our search for neuroprotective agents of microbial origin against excitatory neurotoxins, we have isolated two new bicyclohexapeptides, neuroprotectins A and B, together with a known compound complestatin, from the fermentation broth of Streptomyces sp. Q27107. Neuroprotectins protected primary cultured chick telencephalic neurons from glutamate- and kainate-induced excitotoxicities in a dose-dependant fashion.

Animals↗

Neuroprotectins A and B, bicyclohexapeptides protecting chick telencephalic neurons from excitotoxicity. II. Structure determination.

In the course of our search for neuroprotective agents of microbial origin against kainate-induced neurotoxicity, we have succeeded in the isolation of two new bicyclohexapeptides, neuroprotectins A and B, together with a known compound, complestatin, from the fermentation broth of Streptomyces sp. Q27107. They are closely related in structure to complestatin and possess an oxindolylalanine moiety in place of the tryptophan residue present in complestatin.

Molecular Structure↗

[Development of three-dimensional stereo viewer for high-resolution data].

In order to visualize high-resolution three-dimensional (3D) data as a stereogram, a real-time volume-rendering system using a hardware graphic board and conventional PC was developed. A 256(3) data set could be visualized at a redrawing rate of 12 Hz, and a 512(3) data set at a rate of 2.5 Hz. It was demonstrated that stereogram visualization using volume graphic hardware architecture potentially enables rapid examination of high-resolution 3D data by changing visualization parameters such as level, window, transfer function for opacity, and color map or coordinate direction.

Computer Graphics↗

[Examination of validity of the subjective fatigue scale for young adults].

OBJECTIVE: The purpose of this study was to examine the validity of a new type of Subjective Fatigue Scale for young adults (SFS-Y). METHODS: The SFS-Y consisted of 24 item questions representing 6 sub-scales of difficulty of concentrated thinking, languor, reduced activation, reduced motivation, drowsiness and feeling of physical disintegration. The SFS-Y, subjective symptoms index (SSI), chronic fatigue symptoms index (CFSI), multidimensional fatigue index (MFI) and the Chalder fatigue scale (CFS) were administered to 5435 students aged 15-20 yr. RESULTS: It was inferred that SFS-Y can evaluate individual differences from the viewpoints of score distribution and discrimination power. It was considered that the SFS-Y has high generality because it includes many question items with a similar content to existing fatigue scales and it can do multiple evaluations covering almost all aspects of existing scales. The relationships between sub-scales in the SFS-Y and existing fatigue scales were high in sub-scales with similar names and was low in ones with different names. CONCLUSION: The Subjective Fatigue Scale developed to evaluate subjective fatigue for youths was considered to be excellent in validity and to be effective scale.

Adolescent↗

Up-regulation of cell surface sodium channels by cyclosporin A, FK506, and rapamycin in adrenal chromaffin cells.

Treatment of cultured bovine adrenal chromaffin cells with cyclosporin A (CsA) increased cell surface [(3)H]saxitoxin ([(3)H]STX) binding by 56% in a time (t(1/2) = 15.2 h)- and concentration (EC(50) = 2.9 microM)-dependent manner but did not change the K(d) value. In CsA-treated cells, veratridine-induced (22)Na(+) influx was augmented with no change in the EC(50) of veratridine; also, alpha- and beta-scorpion venom and Ptychodiscus brevis toxin-3 enhanced veratridine-induced (22)Na(+) influx in a more than additive manner, as in nontreated cells. CsA treatment for 1 to 24 h inhibited calcineurin activity, measured by the in vitro assay, with the IC(50) of 0.6 microM but did not alter cellular level of calcineurin. FK506 or rapamycin elevated [(3)H]STX binding by 36 or 25%, whereas GPI-1046, an immunophilin ligand incapable to inhibit calcineurin, or okadaic acid, an inhibitor of protein phosphatases 1 and 2A, had no increasing effect. The rise of [(3)H]STX binding by CsA was attenuated by the coincident treatment with brefeldin A (BFA), an inhibitor of vesicular exit from the trans-Golgi network. The internalization rate of cell surface Na(+) channels, as determined in the presence of BFA, was decreased in CsA (but not rapamycin)-treated cells (t(1/2) = 20.3 h), compared with nontreated cells (t(1/2) = 13.7 h). CsA treatment, however, did not elevate cellular levels of Na(+) channel alpha-subunit and Na(+) channel alpha- and beta(1)-subunit mRNAs. In CsA-treated cells, veratridine-induced (45)Ca(2+) influx via voltage-dependent Ca(2+) channels and catecholamine secretion were enhanced, whereas high K(+)-induced (45)Ca(+) influx was not. Thus, the inhibition of calcineurin or rapamycin-binding protein causes up-regulation of cell surface functional Na(+) channels via modulating externalization and internalization of Na(+) channels, thus enhancing Ca(2+) channel gating and catecholamine secretion.

Adrenal Glands↗

[Optimal setting of focus points for myocardial contrast echocardiography with intravenous Optison (FS-69): an experimental study].

OBJECTIVES: Myocardial contrast echocardiography is useful to assess myocardil perfusion, but myocardial opacification is affected by the setup of the ultrasonic equipment. The optimal setting of focus points for adequate opacification of the myocardium was examined in myocardial contrast echocardiography. METHODS: Myocardial contrast echocardiography was performed in six dogs using triggered second harmonic technology following intravenous administration of Optison(FS-69). The short-axis view was recorded and baseline subtracted video intensity(peak intensity: PI) was calculated at three regions of the left ventricular wall, the anterior, septum, and posterior walls, to evaluate myocardial opacification. The focus point was set at near(2 cm), middle(4 cm) and far(6 cm)points. The myocardial opacification was evaluated at each focus setting. The effect of the acoustic power was also examined by changing the mechanical index to 1.6, 1.2 or 0.8. RESULTS: Myocardial opacification was recognized at all focus points, and segments near the focus points had high PI. However, the PI of the posterior wall was lowest at near focus, whereas the PI of the anterior wall was lowest at far focus. The difference of PI between the anterior and posterior walls was significant (p < 0.05, p < 0.01, respectively) at either focus point. Adequate myocardial opacification of all segments was observed when the focus was set at the middle point. The PI of the whole left ventricle increased relative to mechanical index. CONCLUSIONS: Setting up of focus points at the middle of the left ventricle provides more homogeneous myocardial opacification of the whole left ventricle in myocardial contrast echocardiography, and high acoustic power possibly improves myocardial opacification. Inadequate setting of focus points leads to inadequate estimation of myocardial perfusion by myocardial contrast echocardiography.

Albumins↗

[Usefulness of intravenous myocardial contrast echocardiography in rats].

OBJECTIVES: Few intravenous myocardial contrast echocardiography trials have evaluated myocardial perfusion in small animals. The feasibility of using intravenous myocardial contrast echocardiography to assess the ischemic area in rats was investigated. METHODS: Fourteen open chest Wister male rats were examined. Intravenous myocardial contrast echocardiography was performed by fundamental and intermittent mode using a high frequency (5-12 MHz) transducer (SONOS 5500) with injection of NC100100 (20% dilution) into the femoral vein. The mechanical index was set to 1.6. Baseline-subtracted video intensity (256 level) was measured in the anterior, posterior, septal and lateral walls of the left ventricle. The left anterior descending artery was ligated in 16 rats. The area at risk was evaluated by myocardial contrast echocardiography and compared to the area of defect by Evans Blue staining. RESULTS: All wall segments were clearly opacified (anterior 63.8 +/- 24.7, posterior 27.0 +/- 11.0, septal 44.5 +/- 11.6, lateral 52.3 +/- 19.1), although the baseline-subtracted video intensity of the posterior wall was low. The area at risk was clearly observed, and there was a good correlation with the nonperfused area by Evans Blue staining (y = 1.13x-3.54, r = 0.98). CONCLUSIONS: Intravenous myocardial contrast echocardiography can detect the perfusion defect even in rats using a high frequency transducer and suitable setup of equipment.

Animals↗

[Relationship between the anti-metastatic effect of UFT and in vitro chemosensitivity to 5-FU in metastatic tumors from orthotopic implanted colon cancer in nude rats].

We have investigated the correlation between the in vitro chemosensitivity to 5-FU, measured using the collagen gel droplet embedded culture drug sensitivity test (CD-DST), and the anti-tumor effect of UFT, a prodrug of 5-FU, in metastatic tumors from orthotopic implanted colon cancer in nude rats. Human colon cancer cells (KM12SM) were injected into the cecal wall of the nude rats. Five weeks later, the implanted cecal tumors were removed. Oral UFT (a daily dose of 30 mg/kg) was administered postoperatively for four weeks. After the UFT administration period, the lung and lymph nodes were analyzed macroscopically and microscopically. In vitro chemosensitivity to 5-FU in the lung and lymph node metastases was tested using CD-DST, and the enzymatic activities of thymidine synthetase (TS) and dihydropyrimidine dehydrogenase (DPD) in the lung and lymph node metastases were measured. A daily administration of UFT produced an inhibitory effect on lung metastasis compared with the control group. However, there was no difference in the frequency of lymph node metastasis. The inhibition rate produced by 5-FU in CD-DST was significantly higher for lung metastases than for lymph node metastases. There was no difference in the TS and DPD activities between the metastatic tumoral tissues. These results suggest that the organ specificity of the anti-tumor effects of UFT on colon metastases may be determined by CD-DST of 5-FU for individual tumors. The TS and DPD activity in the tumoral tissues may not affect the organ specificity of the anti-tumor effect of UFT on colon metastases.

Animals↗

Bladder transplantation in rats using FK-506.

PURPOSE: We created viable bladder tissue by transplantation with immunosuppression. MATERIALS AND METHODS: For bladder transplantation the bladder of newborn Brown-Norway rats was excised and each was transplanted into a pouch created in the distal omentum of a 5-week-old Lewis rat. In 15 group 1 rats no immunosuppressive agent was used. In 20 group 2 rats 0.6 mg./kg. FK-506 daily were given intramuscularly until a predetermined day of harvest. Recipient rats were sacrificed on day 3, 5, 7 or 14 after bladder transplantation, and the bladder grafts were harvested and formalin fixed. Hematoxylin and eosin staining was done to examine bladder graft survival and the degree of rejection, and immunohistochemical testing was performed for assessing the vesical nervous system. In 5 rats in the control group bladder augmentation was performed by anastomosing the bladder graft to the native bladder. Each augmented bladder was harvested 21 days later for histopathological assessment. RESULTS: Overall bladder graft survival was 96.4%. Each successfully transplanted bladder graft appeared macroscopically as a thin walled cyst. In group 1 all bladder grafts showed rejection with cellular infiltration. In group 2 there was mild rejection in 5 rats and no evidence of rejection in the remaining 15. All group 2 bladder grafts had intact nerve distribution. Bladder augmentation was successful in all 5 cases and the mucosa was normal throughout each augmented bladder. CONCLUSION: Because FK-506 successfully prevents rejection, our technique would appear to have the potential for creating viable bladder tissue that may be used for bladder augmentation in cases of vesical exstrophy or neurogenic bladder.

Animals↗

Excretion of bikunin and its fragments in the urine of patients with renal stones.

PURPOSE: Bikunin is a Kunitz-type protease inhibitor found in serum and urine. It has been implicated in urinary stone formation. This study was designed to investigate the role of urinary bikunin in stone formation. MATERIALS AND METHODS: Urinary concentrations of bikunin were measured in 18 male formers of urinary stones 28 to 74 years old and in 77 healthy controls, including 39 males and 38 females, without urological abnormality. A sensitive competitive solid phase enzyme immunoassay was established for urinary bikunin. Bikunin was also qualitatively assessed by Western blot analysis. RESULTS: The mean urinary bikunin-to-creatinine ratio plus or minus standard deviation in stone formers was significantly elevated compared with that in healthy male and female controls (52.9 +/- 46.0 microg./mg. creatinine versus 28.0 +/- 30.4 and 26.5 +/- 21.7, p = 0.005 and 0.006, respectively). By Western blot analysis all urine samples contained authentic 40 kDa. bikunin species. However, a significantly higher proportion of patients was found to have aberrant 25 kDa. bikunin species compared with controls (10 of 18 or 55.6% versus 15 of 77 or 19.5%, p = 0.002). Experiments on de-glycosylation with chondroitinase ABC, amino acid sequencing of the aberrant bikunin species and calcium oxalate crystal growth inhibition assay demonstrated that the 25 kDa. bikunin fragment was identical to de-glycosylated bikunin and less inhibitory on calcium oxalate crystal growth. CONCLUSIONS: If urinary bikunin is important in the pathogenesis of urolithiasis, its effect is probably attributable to the concentration and degree of glycosylation.

Adult↗