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Biomedical subjects

J Kobayashi

Publications and source records attributed to J Kobayashi.

At least 163 records · Page 9Linked to original sources

Modulation of multidrug resistance by taxuspine C and other taxoids from Japanese yew.

Taxuspine C (1), a new taxoid from the Japanese yew Taxus cuspidata, increasing the cellular accumulation of vincristine (VCR) in multidrug-resistant tumor cells as potent as verapamil enhanced the chemotherapeutic effect of VCR in P388/VCR-bearing mice. When taxuspine C (1) was given i.p. daily at 200 mg/kg with 0.2 mg/kg VCR for 5 days, a treated/control (T/C) value of 138% was obtained. The other new taxoids, taxezopidines G (8) and H (9), from the yew also increased the VCR accumulation as potent as verapamil. These results suggest that some taxoids may be useful for overcoming multidrug resistance in tumor cells.

Animals↗

Sulfated glycoglycerolipid from archaebacterium inhibits eukaryotic DNA polymerase alpha, beta and retroviral reverse transcriptase and affects methyl methanesulfonate cytotoxicity.

A sulfated glycoglycerolipid, 1-O-(6'-sulfo-alpha-D-glucopyranosyl)-2,3-di-O-phytanyl- sn-glycerol (KN-208), a derivative of the polar lipid isolated from an archaebacterium, strongly inhibited DNA polymerase (pol) alpha and pol beta in vitro among 5 eukaryotic DNA polymerases (alpha, beta, gamma, delta, and epsilon). It also inhibited Escherichia coli DNA polymerase I Klenow fragment (E. coli pol I) and human immunodeficiency virus reverse transcriptase (HIV RT). The mode of inhibition of these polymerases was competitive with the DNA template primer and was non-competitive with the substrate dTTP. KN-208 inhibited pol beta most strongly, with a Ki value of 0.05 microM, 10-fold lower than that for pol alpha (0.5 microM) and 60- or 140-fold lower than that for HIV RT (3 microM) or for E. coli pol I (7 microM), respectively. The loss of sulfate on the 6'-position of glucopyranoside of this compound completely abrogated inhibition. However, the hydrophilic part of KN-208, glucose 6-sulfate alone, showed no inhibition. Other sulfated compounds containing different hydrophobic structures, such as dodecyl sulfate and cholesterol sulfate, exhibited a much weaker inhibition. Our results suggest that the whole molecular structure of KN-208 is required for inhibition. KN-208 was shown to be modestly cytotoxic for the human leukemic cell line K562. Interestingly, a subcytotoxic dose of KN-208 increased the sensitivity of the human leukemic cells to an alkylating agent, methyl methanesulfonate, while it did not potentiate the effects of ultraviolet light or of cisplatin.

Archaea↗

Differential contribution of Na+-K+ pump and K+ conductance to the post-tetanic hyperpolarization of the subtypes of tetrodotoxin-resistant C-fibers in the isolated bullfrog sciatic nerve.

The post-tetanic hyperpolarizations (PTHPs) of the fast and slow tetrodotoxin-resistant (TTX-R) C-fibers were recorded from isolated bullfrog sciatic nerves using a modified vaseline gap method. Under control solutions, the monophasic fast and slow TTX-R C-fibers compound action potentials (C-CAPs) increased in amplitude up to 133.9+/-17.9% (mean+/-SD; n = 9) and 167.0+/-41.5% (n = 7), respectively, shortly after tetanic stimulation. The recovery time constants of the fast and slow ones were 66.5 and 101.2 s, respectively. Significant differences between the rates of amplitude augmentation of the fast and slow TTX-R C-CAPs were found for the most pairs of measured points (P < 0.05 or P < 0.01). After application of ouabain solutions, the augmentation of the fast TTX-R C-CAP was inhibited to an insignificant level, while that of the slow TTX-R C-CAP remained significant with the recovery time constant being 90.9 s. After additional application of TEA, the post-tetanic augmentation of the slow TTX-R C-CAP was blocked. These results suggest that Na+-K+ pump contributes to the PTHPs of the fast and slow TTX-R C-fibers and that K+ conductance contributes only to the PTHP of the slow TTX-R C-fibers, indicating differential modulation of the firing rate of each TTX-R C-fibers group.

Action Potentials↗

Endothelial and serum factors which include apolipoprotein A1 tether elastin to smooth muscle cells inducing serine elastase activity via tyrosine kinase-mediated transcription and translation.

We previously reported that serine elastase activity is induced in cultured porcine pulmonary artery (PA) smooth muscle cells (SMC) following serum stimulation by a mechanism involving adhesion of elastin to an elastin binding protein and tyrosine kinase activity. The present study demonstrates that a PA endothelial cell factor also promotes a fourfold increase in elastin adhesion to PA SMC and a twofold increase in serine elastase activity. The mechanism involves tethering of the factor to SMC, since [3H]-elastin pre-incubated with serum or endothelial cell (EC)-conditioned medium or SMC pre-treated with serum accelerates binding of elastin and tyrosine-kinase related elastase activity. The serum factor appears to interact with integrins as elastase induction is partially inhibited by RGD peptides. The elastase-inducing properties of serum could not, however, be attributed to several RGD-containing proteins. While a 120 kD fibronectin fragment partially reproduced the effect, it was not found in the serum fraction containing elastase-inducing activity. Instead, a 27 kD serum protein was enriched by elastin affinity chromatography, identified as apolipoprotein (Apo) A1 by microsequence analysis, and found to have about 50% of the elastase-inducing activity of serum. Elastase induction is inhibited by actinomycin and cycloheximide, suggesting a requirement for mRNA transcription and protein synthesis. Our results suggest a novel cell-extracellular matrix interaction whereby a soluble factor, in this case a lipoprotein, binds and tethers a matrix component to the cell surface and induces tyrosine kinase-dependent transcription of mRNA culminating in substrate proteolysis.

Animals↗

Rapid detection of male-specific DNA sequence in bovine embryos using fluorescence in situ hybridization.

An accurate, reliable, and quick (less than an hour) method for determining the sex of bovine embryos was developed using a fluorescence in situ hybridization (FISH), with a probe designed from a bovine Y chromosome specific DNA (BC1.2). First, to improve a protocol of FISH and evaluate an accuracy of the method, lymphocyte nuclei prepared from three bulls, two cows, and one freemartin were tested. We found that 5 min was enough for hybridization. The washing solution adequate for posthybridization was 0.5x SSC at 72 degrees C for 5 min. The whole procedure for FISH can be accomplished in less than an hour. A male-specific signal was detected, on average, as 97, 0.5, and 83%, respectively, of lymphocytes in males, females, and a freemartin. Using the rapid FISH protocol developed, 28 embryos were divided. According to the presence of the digoxigenin signal, 16 embryos (57.1%) were predicted as male, and 12 embryos (42.9%), predicted as female.

Animals↗

The effects of FR167653 in extended liver resection with ischemia in dogs.

Interleukin-1 (IL-1) and tumor necrosis factor (TNF) are cytokines commonly associated with inflammatory conditions such as hepatic injury after ischemia-reperfusion. FR167653 has been characterized as a potent suppressant of IL-1beta and TNF-alpha production. In this study, we evaluated the effect of FR167653 in an extended liver resection with ischemia in a dog model. The right portal pedicle was clamped for 60 minutes, while the left portal branch was patent to avoid portal congestion. Following reperfusion, 75% of the liver (including the right central, quadrate, left central, left lateral, and papillary lobes) were resected. Animals were divided into two groups: a control group (n = 10), and a FR-treated group (n = 6) in which FR167653 was administered via the portal vein. Hepatic venous blood was collected to measure alanine transaminase (ALT), aspartate transaminase (AST), lactate dehydrogenase (LDH), purine nucleoside phosphorylase (PNP), and hyaluronic acid (HA) levels, and IL-1beta expression was also measured by reverse-transcriptase polymerase chain reaction (RT-PCR). ALT, AST, LDH, PNP, and HA levels after reperfusion were significantly lower (P < .05) in the FR-treated group than in the control group, and the FR-treated group showed inhibited IL-1beta expression. Liver tissue blood flow, measured by a laser Doppler flow meter, was kept higher in the FR-treated group than in the control group. Histologically, tissue damage was mild in the FR-treated group. The 2-day survival rate was statistically better (P < .05) in the FR-treated group than in the control group. We conclude that FR167653 provides a protective effect for liver parenchyma and sinusoidal endothelial cells in extended liver resection with ischemia.

Alanine Transaminase↗

[Surgical treatment for mitral regurgitation associated with secundum atrial septal defect].

We reviewed 25 patients who underwent a mitral valvuloplasty, from 1984 to 1996, for mitral regurgitation (MR) associated with atrial septal defect (ASD). Mean grade of MR was 2.3 +/- 0.7. The locations of mitral valve lesion were as follows; Postero-medial side of the anterior leaflet (AML) (11 patients: 44%), posteromedial side to center of the AML (7 patients: 28%), whole of the AML (5 patients: 20%), center of the AML (1 patient: 4%), posteromedial side of the posterior leaflet (PML) (1 patient: 4%). In summary, the mitral valve lesion was located in the AML in 96% patients and were seen in the postero-medial side of 96% patients. Mitral valve repair was performed as follows; chordae shortening only (3 patients: 12%), chordae shortening + Kay's annuloplasty (9 patients: 36%), Kay's anuloplasty (10 patients: 40%), using artificial chordae only (1 patient: 4%), using artificial chordae + Kay's annuloplasty (1 patient: 4%), using artificial chordae + ring annuloplasty (1 patient: 4%). In 24 patients, the grade of MR was less than 2/4 in the early postoperative period. In one patient, the grade of 3/4 MR was still remained. Reoperation were required in 2 patients, because of gradual increase of MR, 9 years and 10 years after the initial operation, respectively. In another patient, the grade 3/4 MR recurrently occurred at 6 months after the operation. He has been well maintained medically. In all 4 patients who had more than the grade 3/4 MR postoperatively, the annuloplasty was performed with Kay's method and the cause of MR was poor coaptation around the center of the AML. The mitral valve lesion associated with ASD seemed to be the dislocation of the AML which cause the discrepancy of the coaptation zone between both leaflets, without any prominent prolapse and chordae elongation. We put a particular emphasis on that the mitral valve repair should be performed with the recognition of the etiology of the mitral valve lesion. Especially, if the lesion extends around the center of the AML, sufficient coaptation area of both leaflets at the center of the AML should be obtained by anuloplasty.

Adult↗

Intramucosal pH and intestinal mucosal damage in ischemia-reperfusion injury.

Small bowel transplantation (SBT) has become an increasingly promising treatment for short bowel syndrome. The evaluation of graft viability after SBT, however, has not been established, except by mucosal biopsy. We monitored intestinal mucosal acidity in order to detect small intestinal ischemia-reperfusion injury. Mongrel dogs were used in this study. After laparotomy, the small bowel was isolated with a vascular pedicle. A tonometer to measure intramucosal pH (pHi) was then positioned in the terminal ileum. The superior mesenteric artery was occluded with or without concomitant superior mesenteric vein occlusion for 60 or 120 min. The value of pHi was determined from laparotomy (baseline) to 12 h after reperfusion. Whole-thickness specimens of the ileum were taken before ischemia, just before reperfusion, and 1 h afterward. Mucosal injury was graded histopathologically. pHi decreased from baseline in relation to the degree of histopathological mucosal injury. There was a significant correlation between histological findings and the change in pHi. We conclude that monitoring intestinal mucosal acidity is a reliable way of determining graft viability after SBT.

Animals↗

Anastomosis of the left juxtaposed atrial appendages in a patient with tricuspid atresia.

A 9-month-old boy with left juxtaposition of the atrial appendages, tricuspid atresia, pulmonary atresia, and ventriculoarterial discordance underwent anastomosis between the atrial appendages after failure of balloon/blade atrial septostomy because of restrictive atrial septal defect. For surgical creation of atrial communication in patients with juxtaposed atrial appendages, anastomosis between the atrial appendages seemed to be safer, more effective, and less invasive than septectomy by Blalock-Hanlon technique or inflow occlusion technique.

Anastomosis, Surgical↗