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Biomedical subjects

T Wakabayashi

Publications and source records attributed to T Wakabayashi.

At least 145 records · Page 8Linked to original sources

[Studies on 5-lipoxygenase inhibitors. Synthesis of and structure--activity relationship in p-hydroxyarylalkenylbenzoazoles].

In a previous paper we reported that 2-(p-hydroxyarylbutadienyl)benzoxazoles are highly potent 5-lipoxygenase inhibitors. We synthesized their ethenyl homologues and benzothiazole derivatives, and evaluated their 5-lipoxygenase inhibitory activity in vitro with cell-free rat basophilic leukemia (RBL-1). In most cases the replacement of benzoxazolyl with benzothiazolyl resulted in an enhancement of the activity. All compounds with butadienyl spacers tested herein exhibited strong inhibitory activities. While most of the ethenyl homologues showed weaker activities than their corresponding butadienyl homologues, some ethenyl compounds in the benzothiazole derivatives were found to be as potent as their corresponding butadienyl homologues. The inhibitory activity was also affected by the variation in the p-hydroxyaryl functionality.

Animals↗

Abducens nerve neurinoma--case report.

A 61-year-old female presented with an abducens nerve neurinoma manifesting as diplopia and facial numbness. The tumor was totally removed, and the exact origin was confirmed intraoperatively. Neuroradiological imaging and clinical history are frequently insufficient to obtain an accurate preoperative diagnosis of intracranial neurinoma.

Abducens Nerve↗

Enzyme immunoassay of glioma cell tenascin secretion and augmentation by tumor necrosis factor-alpha.

Expression of tenascin, an extracellular matrix glycoprotein, was measured in glioma cell lines using a newly established enzyme immunoassay. Secreted tenascin was found at concentrations greater than 800 ng/ml in eight of 14 glioma, three small cell lung carcinoma, two melanoma, and one sarcoma cell lines. The remaining six glioma and other carcinoma cell lines, and cell lines originating from normal tissues demonstrated low levels or no secretion into the supernatant. The glioma cell line, U-251-MG nu/nu, which has almost 100% transplantability in nude mice, had the highest expression level of tenascin among the glioma cell lines examined. Even low secretor glioma cell lines released high concentrations of tenascin, detectable by assaying the NP-40 solubilized cell lysates. Flow cytometric analysis revealed that tenascin was located on both the cell surface and primarily in the cytoplasm of glioma cells. When the glioma cell lines were exposed to tumor necrosis factor-alpha (TNF-alpha), levels of secreted tenascin increased between 36% and 380%, whereas transforming growth factor-beta induced only minimal changes. These results suggest that glioma cell lines may be classified according to the degree of tenascin secretion/expression: high secretor type, low secretor type, and non-expressing type. The increase in tenascin secretion by TNF-alpha suggests that the expression of tenascin in glioma growth and development may be mediated through a cytokine network.

Animals↗

Antitumor activity of recombinant human tumor necrosis factor-alpha (rH-TNF alpha) and liposome-entrapped rH-TNF alpha.

The antitumor activities of recombinant human tumor necrosis factor-alpha (rH-TNF alpha) and liposome-entrapped rH-TNF alpha were evaluated in various glioma cell lines and a rat brain T9 gliosarcoma model. rH-TNF alpha had a direct cytotoxic activity against various glioma cell lines in vitro, and indirect cytotoxic activity against gliosarcoma (T9) in vivo. Liposome-entrapped rH-TNF alpha had increased direct cytotoxic activity in vitro, and against experimentally induced brain tumors in vivo. The effects in vivo were probably due to vascular damage of the tumor vessels as shown by histological examination and activation of cytotoxic macrophages as shown in vitro. These results indicate that the general or local administration of liposome-entrapped rH-TNF alpha may become a useful adjunct treatment for malignant brain tumor.

Animals↗

[Measurement of relative plasma volume and plasma refilling rate during ultrafiltration of hemodialysis by measuring apparatus of plasma colloidal osmotic pressure].

BACKGROUND: The objective of this study is the prevention of unexpected hypotension during hemodialysis caused by unsuitable filtration rate. METHODS: The plasma colloidal osmotic pressure (COP) was measured continuously during ultra-filtration, and relative plasma volume (%PV) and plasma refilling rate (PRR) were calculated during 12 ECUMs and one dialysis on twelve patients in early stage of chronic renal failure. Values of %PV and PRR calculated from COP were drawn in graphic curves, and analysed to obtain characteristic pattern. Minimum value of %PV (%PV min), maximum value of PRR (PRRmax) and time for PRR to decrease to 95% (95% PVt) on the curves were documented simultaneously. RESULTS: At the initial stage of %PV curve, obvious fall was observed on 6 of 13 estimation (initial fall of %PV). On the contrary at the initial stage of PRR curve, obvious rise was observed on 5 estimation (initial rise of PRR). A close relationship was indicated between the two phenomenons. In 6 cases with initial fall of %PV and in 5 cases with initial rise of PRR body weight, PRRmax and 95%PVt were lower than another cases without it. The ultra-infiltration velocity was estimated to be relatively high in these groups. In these 8 cases, filtrated water volume was judged as adequate clinically. In 4 of 5 cases without final fall of PRR, it was judged as inadequate and needed to evacuate more 0.9 to 3.0 kg (average of 1.8 +/- 1.07) of water from the patients. CONCLUSIONS: From the above, we concluded that our method is useful for deciding suitable velocity of ultrafiltration and dry weight in hemodialysis therapy.)

Adult↗

Borrelidin is an angiogenesis inhibitor; disruption of angiogenic capillary vessels in a rat aorta matrix culture model.

Borrelidin, an antibiotic from Streptomyces rochei, was found to be an angiogenesis inhibitor in a rat aorta matrix culture model which forms capillary vessels in vitro. Borrelidin strongly inhibited capillary tube formation with a 50%-inhibitory concentration value of 0.8 nM, and decreased the number of capillary tubes within 24 hours when added after maturation of tube formation. Borrelidin remarkably disrupted capillary tubes in a dose-dependent manner, by inducing apoptosis of the tube-forming cells.

Animals↗

[The studies of extracorporeal shock wave lithotripsy (ESWL) for pancreatic ductal stones].

30 patients with main pancreatic duct stones were treated by ESWL. In 18 of 22 patients who had not previously undergone endoscopic pancreatic sphincterotomy (EPST) or endoscopic sphincterotomy (EST), the stone fragments disappeared after ESWL. The fragments were removed endoscopically in the remaining 4 cases. Complete clearance was achieved in 8 cases with endoscopically unextractable stones by ESWL. After the ESWL procedure, absolute relief from pain was reported by in 19 of 22 patients with abdominal complaints. Serum amylase levels decreased significantly, and dilatation of the main pancreatic duct (MPD) was reduced. In the medium-term follow-up period, pancreatic exocrine function and endocrine function had a possibility to be preserved. One case of pancreatic cancer and one case of an intraductal papillary tumor of the pancreas were found, indicating that careful observation is necessary even after complete removal of pancreatic stones. In cases of Santorini duct dominant, multiple stones, or stricture of the MPD, ESWL should be combined with EPST and endoscopic stenting for preventing recurrence of acute pancreatitis and pancreatic stones. In conclusion, ESWL is the first choice of treatment for pancreatolithiasis and useful procedure and the limited complications.

Adult↗

Genetic profiles of newly inbred Dahl/Iwai salt-sensitive and salt-resistant rats.

Dahl-Iwai salt-sensitive (S) and salt-resistant (R) rat strains were established as inbred strains at Brookhaven National Laboratory, NY, and were introduced into Eisai Co., Ltd., Japan, and designated DIS/Eis and DIR/Eis. To examine whether there are different allele distributions among the substrains of inbred Dahl S and R rats, we determined biochemical and immunological alleles of DIS/Eis and DIR/Eis, and SS/Sea and SR/Sea, which were derived from SS/Jr and SR/Jr, which were developed by Rapp and Dene. Several differences of allele distribution were observed, indicating that the substrains have different genetic backgrounds. The phenotypic differences between the substrains, such as the severity of the hypertension induced, could be ascribed to the different genetic backgrounds.

Alleles↗

New form of platyspondylic lethal chondrodysplasia.

We report on a sporadic case of hitherto unknown lethal skeletal dysplasia. The cardinal clinical manifestations consisted of frontal bossing, cloudy corneae, low nasal ridge, and micrognathia, hypoplastic thorax, and rhizomelic micromelia. Laryngoscopy and neck CT disclosed laryngeal stenosis, and brain CT demonstrated hypoplasia of the corpus callosum. Skeletal survey demonstrated hypoplasia of facial bones and short skull base, extremely severe platyspondyly, hypoplastic ilia, and delayed epiphyseal ossification and rhizomelic shortness of tubular bones. The long bones appeared overtubulated with exaggerated metaphyseal flaring. The humeri were particularly short and bowed. Bowing of the radii and ulnae with subluxation of radial heads presented as a Madelung-like deformity. Unlike the long bones, the short tubular bones were not short and normally modeled. The skeletal changes were superficially similar to those in a group of lethal platyspondylic chondrodysplasias, but were inconsistent with any known subtypes of this group or other lethal skeletal dysplasias.

Abnormalities, Multiple↗

Tropomyosin-binding site(s) on the Dictyostelium actin surface as identified by site-directed mutagenesis.

To identify tropomyosin-binding site(s) on the surface of actin molecule, we examined the effect of mutagenesis introduced to subdomain 4 of actin. Because the sequence of Gln228-Ser232 of Dictyostelium actin differs from that of Tetrahymena actin that does not bind tropomyosin, the Dictyostelium/Tetrahymena chimeric actin was produced. Also, Lys238 and Glu241 were replaced with alanine (mutant 645) to study the role of charged residues which are located at both ends of a beta-sheet. As a control experiment, a negative charge was introduced near to the N-terminus (mutant 663). To facilitate the separation of mutant actins without affecting the normal function, Glu360 was replaced with histidine. As a control mutant to such mutants, the mutant 647 (E360H) was produced. Mutant actins were expressed in Dictyostelium cells. All mutant actins were functional: they (i) polymerize and (ii) activate ATPase activity of rabbit skeletal myosin subfragment-1 (S1). The mutant 663 (G2E) showed tropomyosin binding and activated myosin ATPase almost as well as rabbit skeletal actin. However, the tropomyosin binding of the mutant 645 (K238A/E241A/E360H) became magnesium dependent. The chimeric actin (mutant 646: QTAAS-to-KAYKE replacement and E360H) showed decreased tropomyosin binding even in the presence of magnesium ions. These results indicate that the tropomyosin-binding sites of "on"-state actin are on subdomain 4. Surprisingly, the chimeric actin showed more cooperative calcium regulation than rabbit skeletal actin in the presence of tropomyosin-troponin. The mutant actin 645 can hardly activate S1 ATPase irrespective of calcium concentration in the presence of tropomyosin-troponin, even though this actin by itself can activate S1 ATPase. The steric blocking or cooperative/allosteric mechanism of thin filament regulation is discussed.

Actins↗

Monoclonal antibody C38 labels surviving retinal ganglion cells after peripheral nerve graft in axotomized rat retina.

C38 is a monoclonal antibody that labels retinal ganglion cells in both intact and axotomized rat retinas. We report here that C38 labeled retinal ganglion cells that survived after optic nerve section and peripheral nerve graft in rats. Furthermore, with combination of the retrograde labeling, we succeeded to distinguish surviving retinal ganglion cells without axonal regeneration from those with regenerating axon.

Animals↗

Suppression of CD44 expression decreases migration and invasion of human glioma cells.

We have reported high expression of CD44H in human glioma cells. To investigate the role of CD44H in the invasion of human glioma, we established a CD44-anti-sense-gene-expression glioma cell line named U-251A1. The expression of CD44H in the G-418-selected U-251A1 cells was reduced to 20% of that in the parental U-251SP cells, as determined by flow-cytometry analysis. We first examined the migratory responses of U-251A1 cells in vitro by time-lapse video-microscopic sparse cell-migration assay on hyaluronic acid or on chondroitin 6 sulfate. U-251A1 cells did not show significant differences in motility on any substrate, while U-251SP and other CD44H-positive cells showed dose-dependent increase of migration specifically on hyaluronic acid. To examine the physiologic function of CD44H in gliomas in vivo, U-251A1 and its control cells, U-251S1, which retain CD44-sense-expression vector, were injected stereotactically into the brains of nude mice. U-251A1 cells were localised in the region of the injection site, with relatively well demarcated borders between tumour and brain tissue, while the control cells demonstrated a cell-infiltration pattern. Our data suggest that CD44H may be required for infiltration of glioma cells through its interaction with hyaluronic acid, a major component of the brain extracellular matrix.

Animals↗

Monoclonal antibody C38 recognizes retinal ganglion cells in cats and rats.

We developed monoclonal antibody C38 which specifically recognizes retinal ganglion cells (RGCs) in flatmount preparations of cat and rat retinas. We first induced immunological tolerance in Balb/c mice against axotomized rat retinas which lack most of the RGCs. Then the mice were immunized with intact rat retinas to produce antibodies against RGCs. Monoclonal antibody C38 appeared to be specific for cat RGCs based on immunoreactivities seen in flatmounts and vertical sections of the retina. In rats, we verified that over 90% of retrogradely labeled RGCs were immunoreactive for C38 antibody. In axotomized rat retinas, surviving RGCs were labeled with C38 without erroneous labeling of glial cells. The antigen that C38 recognized was 24 kDa in molecular weight and found in cerebrum, cerebellum, and spinal cord as well as retina. It is suggested that monoclonal antibody C38 is a useful label for RGCs.

Animals↗

Mechanism of the formation of megamitochondria in the mouse liver induced by chloramphenicol.

Correlation between chloramphenicol-induced formation of megamitochondria in the mouse liver and oxidative stress was studied by lipid peroxidation analysis and electron microscopic technique. Chloramphenicol suppressed increases in the body weight and liver weight of experimental animals and at the same time induced a remarkable increase in lipid peroxidation in the liver during the formation of megamitochondria. A spin trapping agent, 4-hydroxy-2,2,6,6-tetramethyl-piperidine-1-oxyl, abolished all these changes induced by chloramphenicol. Namely, both the body weight and liver weight of chloramphenicol-treated animals stayed at the same levels as those of the control, and the formation of megamitochondria was completely suppressed. Allopurinol, a xanthine oxidase (EC 1.2.3.2) inhibitor, partly inhibited the changes induced by chloramphenicol, as described above. These results suggest that chloramphenicol-induced formation of megamitochondria is not simply ascribed to the suppression of the dividing process of mitochondria due to lowered protein synthesis in mitochondria but is intimately related to oxidative stress. Furthermore, the results obtained with allopurinol may indicate that enhanced levels of lipid peroxidation observed in chloramphenicol-treated animals are partly due to enhanced rate of the degradation of purine nucleotides catalyzed by xanthine oxidase.

Allopurinol↗

Severe hypertension and cardiac failure associated with neuroblastoma: a case report.

The authors report on 3-year-old-girl with neuroblastoma complicated by severe hypertension and cardiac failure. She had cardiomegaly and pleural and pericardial effusions. Echocardiogram showed left ventricular hypertrophy and decrease of the left ventricular ejection fraction to 0.36 (normal > .40). Abdominal computed tomographic scan indicated a 7 x 7-cm tumor in the left suprarenal area. There was a marked increase in catecholamines and metabolites in her body fluids. After hypertension was controlled with doxazosin (a long-acting alpha 1 adrenergic blocker), her cardiac function gradually improved. A tumor was surgically removed and diagnosed as a poorly differentiated ganglioneuroblastoma. Preoperative differentiation between neuroblastoma and pheochromocytoma was not possible on the basis of catecholamine analysis or imaging studies including computed tomography scan and magnetic resonance imaging. It is important to control hypertension quickly in the patients with catecholamine-induced cardiomyopathy to facilitate surgical intervention for diagnosis and treatment.

Adrenergic alpha-Antagonists↗

New approach for the treatment of medulloblastoma by transfection with glial fibrillary acidic protein gene.

Glial fibrillary acidic protein (GFAP) is one of the intermediate filaments found in mature normal astrocytes and differentiated glioma cells. It seems to be able to stabilize the cytoskeleton of the astrocyte and may play a role in maintaining astrocyte cell shape, in association with other cytoskeletal components such as microfilaments and microtubles. However, its tissue-specificity remains unclear. To clarify the effect of GFAP expression in brain tumour cells, transfer of the GFAP gene into the human medulloblastoma cell line, DAOY-1 (which does not express GFAP) was carried out using liposomes. Upon transfection, we observed the alterations in the characteristics of GFAP transfected cells. Cell growth, morphology and sensitivity to anticancer drugs were compared between GFAP gene transfected DAOY-1 and control DAOY-1 cells. Growth inhibition and increase of sensitivity to anticancer drugs were observed with GFAP expression in GFAP gene-transfected DAOY-1 cells. However, no morphological changes were noted.

Brain Neoplasms↗

Detection of K-ras point mutations at codon 12 in pure pancreatic juice for the diagnosis of pancreatic cancer by PCR-RFLP analysis.

The present study was undertaken to detect K-ras point mutations at codon 12 in pure pancreatic juice (PPJ) for the diagnosis of pancreatic cancer (PC) using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. PPJ was collected through a cannula under a duodenal fiberscope from 26 patients with PC and 32 patients with chronic pancreatitis (CP). DNA was extracted from PPJ and was used as the template for PCR. Analysis of PPJ by PCR-RFLP with BstNI revealed that the incidence of K-ras point mutations at codon 12 was 81% (21/26) in patients with PC and 6% (2/32) in those with CP. With reference to the location of PC, the incidence of K-ras mutations was 79% (11/14) in the head, 86% (6/7) in the body, and 80% (4/5) in the tail of the pancreas. The incidence of K-ras mutants was 50% (1/2) in tumor size 1 (TS1; < or = 2.0 cm in size), 71% (5/7) in TS2 (2.1 to < or = 4.0 cm), 89% (8/9) in TS3 (4.1 to < or = 6.0 cm), and 88% (7/8) in TS4 (> 6.1 cm). These results suggested that analysis of K-ras point mutations at codon 12 in PPJ using the PCR-RFLP method is a promising new genetic test for the diagnosis of PC.

Adult↗