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Biomedical subjects

M Sugimoto

Publications and source records attributed to M Sugimoto.

At least 73 records · Page 4Linked to original sources
PubMed2000

Proteomic analysis of Epstein-Barr virus-transformed human B-lymphoblastoid cell lines before and after immortalization.

Infection of human B lymphocytes with Epstein-Barr virus (EBV) induces proliferative B-lymphoblastoid cell lines (LCLs). However, the majority of EBV-transformed LCLs are mortal and unable to avoid cellular senescence. In our previous experiment, three immortalized LCLs were established by passages of EBV-transformed LCLs for nearly five years accompanied by strong telomerase activity. In the present study, proteomic profiles of these three LCLs were analyzed comparatively at the early and the late passages of cell culture, and a protein spot was found which most significantly decreased with the immortalization in two LCLs. The expression of the protein in the third LCL was suppressed at 17 population doubling level (PDL), already suggesting that part of the immortalization process had been initiated before 17 PDL. The protein was assigned to ssp7001 (16.3 kDa, pI 6.0) by referring to our TMIG-2DPAGE proteome database. The protein was transferred onto a polyvinylidene difluoride (PVDF) membrane and digested with lysilendopeptidase to perform peptide mass fingerprinting by nanoelectrospray ionization mass spectrometry (nano-ESI-MS). Subsequent MS-Fit database search indicated that ssp7001 is a phosphoprotein stathmin. This speculation was confirmed by the tandem MS (MS/MS) analysis in a Q-Tof system and by Edman degradation microsequencing.

Amino Acid Sequence↗
PubMed2000

Apoptosis in skin pigment cells of the medaka, Oryzias latipes (Teleostei), during long-term chromatic adaptation: the role of sympathetic innervation.

Many teleost fish can adapt their body color to a background color by changing the morphology and density of their skin pigment cells. Melanophore density in fish skin decreases during long-term adaptation to a white background. Although cell death, especially apoptosis, is thought to be involved in these morphological changes, there are no data clearly supporting this mechanism. Using medaka fish, Oryzias latipes, we observed that, on a white background, melanophore size was reduced first and this was followed by a decrease in melanophore density caused by gradual cell death. The process of cell death included loss of cell activity, cell fragmentation, phagocytosis of the fragments, and clearance via the epidermis. Apoptosis was assessed by the appearance of phosphatidylserine on the cell surface of melanophores that had lost motile activity, and DNA fragments involved in cell fragmentation were detected by the TUNEL (TDT-mediated dUTP-biotin nick end-labeling) assay. However, when chemically denervated fish were used, although melanophore size was reduced as expected, cell death was suppressed even on a white background. In skin tissue culture, apoptosis in melanophores was stimulated significantly by norepinephrine, but not by melanin-concentrating hormone. These results indicate that melanophore density decreases by apoptosis, and suggest that sympathetic innervation has an important role in the regulation of apoptosis in melanophores. In analogous fashion, leucophores showed a significant decrease in density with an increase of cell death on a black background. We suggest that apoptosis regulates the balance of pigment cells in the skin of medaka fish to adapt their body color to a particular background.

Adaptation, Physiological↗
PubMed2000

Attachment of bacterial cells to carbon electrodes.

Anodic stripping method was applied to analyze the process of bacterial attachment to the surface of carbon-paste electrodes (CPE). The electrode was immersed for various times in a bacterial cell suspension to allow the cells to attach to its surface. The number of bacterial cells attached to the electrode surface increased along with time. On the other hand, the current derived from the oxidation of a dye, Hoechst, which was adsorbed to the surface after attaching the bacterial cells, decreased along with time. It was considered that the current output, correlated with the amount of dye, adsorbed onto regions where no bacterial cell attached. These results indicate that the bacterial-attachment process can be analyzed by measuring the electric current derived from the dye instead of counting the number of attached cells.

Bacterial Adhesion↗
PubMed2000

Participation of the conjugated diene part for potent cytotoxicity of callystatin A, a spongean polyketide.

5-epi, 10-epi, 8-Deethyl, and 10-demethyl analogues of callystatin A, a potent cytotoxic spongean polyketide, were synthesized to elucidate structure-requirement for cytotoxic potency. Inversion of the asymmetric center at C-10 in callystatin A minimally affected the activity, while lack of the 10-methyl group in callystatin A decreased cytotoxicity. In addition, the C-5 epimer and the 8-deethyl analogue of callystatin A showed weaker cytotoxicity.

Animals↗
PubMed2000

Production of 105Rh-EDTMP and its bone accumulation.

105Rh has favorable physical characteristics as a radiotherapeutic nuclide. Carrier-free 105Rh can be produced by the neutron activation of 104Ru followed by beta decay of 105Ru. It was clarified that carrier-free 105Rh can be produced in quantities and the purity necessary for chemical and clinical investigations of its use as a nuclide for radiotherapy. 105Rh-EDTMP was simply obtained from 105Rh3+ and EDTMP by heating for 30 min in boiling water, giving a radiochemical yield of > 99%. Dissociation of radioactivity assessed by paperchromatography was negligible for up to 5 days after its preparation. In animals, 105Rh-EDTMP showed rapid blood clearance and selective uptake in the bone. Hence, 105Rh-EDTMP is thought to be a promising therapeutic agent for the treatment of pain due to bone metastases.

Animals↗
PubMed2000

Potent in vivo antimalarial activity of 3,15-di-O-acetylbruceolide against Plasmodium berghei infection in mice.

The antimalarial activity of the O-acylated bruceolide derivative, 3,15-di-O-acetylbruceolide, was evaluated against Plasmodium berghei in vivo. The concentration of 3,15-di-O-acetylbruceolide required for 50% suppression (ED50) of P. berghei in mice was 0.46 +/- 0.06 mg/kg/day, whereas bruceolide was only half as effective as 3,15-di-O-acetylbruceolide. Two antimalarial drugs used clinically, chloroquine and artemisinin, demonstrated only low activity corresponding to 1/4 and 1/12 of the ED50 value of 3,15-di-O-acetylbruceolide, respectively. These results may be helpful in the design of better chemotherapeutic bruceolides against falciparum malaria.

Animals↗
PubMed2000

Effects of the mycelial extract of cultured Cordyceps sinensis on in vivo hepatic energy metabolism and blood flow in dietary hypoferric anaemic mice.

The beneficial effects of a traditional Chinese medicine, Cordyceps sinensis (Cs), on mice with hypoferric anaemia were evaluated by NMR spectroscopy. Experimental hypoferric anaemia was induced in mice by feeding with an Fe-free diet for 6 weeks. They were then given extract from cultured Cs (200 mg/kg body weight daily, orally) and were placed on an Fe-containing recovery diet (35 mg Fe/kg diet) for 4 weeks. In vivo 31P and 2H NMR spectra acquired noninvasively and quantitatively at weekly intervals were used to evaluate hepatic energy metabolism and blood flow in the mice. During the 4-week Cs-extract treatment, consistent increases were observed in liver beta-ATP: inorganic phosphate value by liver 31P NMR spectroscopy, representing the high energy state, and in blood-flow rate as determined by 2H NMR spectroscopy of deuterated water (D2O) uptake after intravenous injection of D2O. The haematological variables (the packed cell volume and the haemoglobin level) and the hepatic intracellular pH, which was determined from the NMR chemical shift difference between the inorganic phosphate peak and the alpha-phosphate peak of ATP, were not significantly different between Cs-extract-treated and control mice. As blood flow and energy metabolism are thought to be linked, the Cs-extract-increased hepatic energy metabolism in the dietary hypoferric anaemic mice was concluded to be due to increased hepatic blood flow.

Adenosine Triphosphate↗
PubMed2000

Cytochalasin A inhibits the binding of phenylalanine ammonia-lyase mRNA to ribosomes during induction of phytoalexin in pea seedlings.

Cytochalasin A (CA) blocked the accumulation of phytoalexin and phenylalanine ammonia-lyase (PAL)-protein in pea tissues treated with a fungal elicitor but scarcely affected the PAL-mRNA content. Further analysis showed that CA decreased the PAL-mRNA bound to ribosomes. These results indicate that actin filaments are tightly associated with the translational process of the PAL gene.

Benzopyrans↗
PubMed2000

Activation of cerebral function by CS-932, a functionally selective M1 partial agonist: neurochemical characterization and pharmacological studies.

A newly synthesized agonist for muscarinic acetylcholine (ACh) receptors CS-932, (R)-3-(3-iso-xazoloxy)-1-azabicyclo-[2.2.2]octane hydrochloride, showed a relatively higher affinity for M1 than M2 receptors expressed in Chinese hamster ovary (CHO)-cells in comparison with ACh. CS-932 elevated the intracellular Ca2+ level only in M1-CHO cells, although ACh increased the level in both M1- and M3-CHO cells. CS-932 and ACh reduced forskolin-stimulated accumulation of cAMP in M2-CHO cells by 20% and 80%, respectively. This neurochemical profile of CS-932 indicates that the compound can activate M1-receptor-mediated functions selectively. CS-932 increased firing of cholinoceptive neurons in rat hippocampal slices, and this excitation was antagonized by pirenzepine, but not by AF-DX 116. CS-932 increased awake and decreased slow wave sleep episodes of daytime EEG in free-moving rats. It counteracted scopolamine-induced slow waves in rat cortical EEG. CS-932 also increased the power of alpha- and beta-waves, but decreased delta-wave of the cortical EEG in anesthetized monkeys. It ameliorated scopolamine-induced impairment of working memory in rats. Orally administered CS-932 had the best penetration into the brain among the muscarinic agonists tested and caused the least salivary secretion among the cholinomimetics examined. These results indicate that CS-932 has potential as a cognitive enhancer with fewer side effects in therapy for Alzheimer disease.

Animals↗
PubMed2000

Surgical repair of a depressed fracture in a green sea turtle, Chelonia mydas.

Sea turtles are considered to be endangered species. A depressed fracture of a 35 kg green sea turtle was treated surgically. Isoflurane was used for induction and maintenance of anesthesia. Slow induction of and slow recovery from anesthesia was remarkable. After the operation, there was an improvement of general status, but head tilt and weakness of the left limbs persisted. As the turtle did not eat, force feeding using stomach tube was performed. The turtle died at about 6 months after the surgery.

Anesthesia, Inhalation↗
PubMed2000

Proficient repair of potentially lethal damage sensitive to hypertonic treatment in osteosarcoma cells.

Fast-repairing potentially lethal damage (PLD) in seven osteosarcoma cell lines was analyzed after treatment with a hypertonic 0.5 M NaCl solution for 20 min and compared to that in seven human fibroblast strains. Fixation of PLD after exposure to ionizing radiation was observed without exception in both the osteosarcoma cells and the fibroblast strains. The percentages by which the D(o)'s of the osteosarcoma cells decreased were significantly higher than the percentage decreases in the C(o)'s of the fibroblast strains (P < 0.01). Hypertonic treatment resulted in radiosensitization due to fixation of PLD in all of the osteosarcoma cell lines, demonstrating that osteosarcoma cells can repair PLD better than normal fibroblast cells. The radiobiological response of the osteosarcoma cells, with enhanced killing after hypertonic treatment, was similar to that of normal untreated fibroblast cells.

Bone Neoplasms↗
PubMed2000

X chromosome inactivation revealed by the X-linked lacZ transgene activity in periimplantation mouse embryos.

Using H253 mouse stock harboring X-linked HMG-lacZ transgene, we examined X chromosome inactivation patterns in sectioned early female embryos. X-gal staining patterns were generally consistent with the paternal X inactivation in the trophectoderm and the primitive endoderm cell lineages and random inactivation in the epiblast lineages. The occurrence of embryonic visceral endoderm cells apparently at variance with the paternal X chromosome inactivation in 7.5 dpc embryos was explained by the replacement of visceral endoderm cells with cells of epiblast origin. The frequency of cells negative for X-gal staining in 4.5-5.5 dpc XmXp* embryos fluctuated considerably especially in the extraembryonic ectoderm and the primitive endoderm, whereas it was less variable in the embryonic ectoderm. We could not, however, determine whether it is a normal phenomenon revealed for the first time by the use of HMG-lacZ transgene or an abnormality caused by the multicopy transgene.

Animals↗
PubMed2000

[A novel in vivo 31P-nuclear magnetic resonance technique for assessment of teratogenicity induced by environmental endocrine disrupters in mice].

Novel in vivo 31P-nuclear magnetic resonance spectoroscopy(NMR) and NMR-imaging techniques for accurate and noninvasive assessment of teratogenicity induced by environmental endocrine disrupters(EDs) were developed. Mice with pregnancy were administered ED at extremely low dose, and then in vivo 31P-NMR spectra of embryos were acquired noninvasively and quantitatively to evaluate the energy metabolism. A significant decrease in embryo ATP level was seen, but no significant changes were detected by conventional histological and biochemical methods. In conclusion, in vivo NMR techniques are highly sensitive(at least 100-fold more sensitive than conventional methods) and are useful for toxicological assessment of environmental pollutants.

Abnormalities, Drug-Induced↗
PubMed2000

Stable earthworm serine proteases: application of the protease function and usefulness of the earthworm autolysate.

The fibrinolytic enzymes from Lumbricus rubellus [Nakajima, N. et al., Biosci. Biotechnol. Biochem., 57, 1726-1730 (1993), 60, 293-300 (1996), and 63, 2031-2033 (1999)] were further characterized to exploit their catalytic functions. These enzymes are stable in solution for long periods at room temperature and strongly resistant to organic solvents, even toluene and n-hexane. The serine proteases can act on various protein substrates such as elastin and hemoglobin as well as fibrin, and also catalyzed the hydrolysis of esters such as ethyl acetate and a bioplastic, poly[(R)-3-hydroxybutyrate] film. The enzymes, in the absence of microbial degradation, contributed to the production of the earthworm autolysate possessing antioxidant ability and protease activity, whose components were similar to those of soy sauce. The extract of the earthworm autolysate could be used as a peptone substitute in media for the cultivation of microorganisms.

Journal Article↗
PubMed1999

Regulation of CDK4 activity by a novel CDK4-binding protein, p34(SEI-1).

The p16(INK4a) tumor suppressor inhibits cyclin-dependent kinases (CDK4 and CDK6). Here we report the isolation of a novel gene, SEI-1, whose product (p34(SEI-1)) appears to antagonize the function of p16(INK4a). Addition of p34(SEI-1) to cyclin D1-CDK4 renders the complex resistant to inhibition by p16(INK4a). Expression of SEI-1 is rapidly induced on addition of serum to quiescent fibroblasts, and ectopic expression of p34(SEI-1) enables fibroblasts to proliferate even in low serum concentrations. p34(SEI-1) seems to act as a growth factor sensor and may facilitate the formation and activation of cyclin D-CDK complexes in the face of inhibitory levels of INK4 proteins.

Animals↗