Search PubMed⌕ Search

Biomedical subjects

M Sakai

Publications and source records attributed to M Sakai.

At least 289 records · Page 16Linked to original sources
PubMed1996

Lectin affinity electrophoretic demonstration of tissue specificity and malignant alteration of human alpha-fetoprotein isoforms produced in transgenic mice.

Transgenic mouse which produces human alpha-fetoprotein (AFP) ubiquitously was used to study carbohydrate structures of AFP produced by various tissues as well as that in the serum. A series of tissues from the transgenic mouse were cultured in vitro and the AFP produced was analyzed by affinity electrophoreses with 4 kinds of lectins. Variable electrophoretic profiles of them suggested that the fine structures of the carbohydrate of human AFPs expressed in the mouse tissues were different. The characterization of human AFP produced by mouse hepatoma which developed in offspring between the AFP transgenic mouse and hepatoma-developing transgenic mouse indicated that the hepatoma AFP was distinct from the liver AFP and that the malignancy-specific phenotypic alteration was common to human and mouse.

Animals↗
PubMed1996

Activation of acyl-coenzyme A:cholesterol acyltransferase activity by cholesterol is not due to altered mRNA levels in HepG2 cells.

Many studies have shown that sterols can stimulate acyl-coenzyme A:cholesterol acyltransferase (ACAT) activity in cells. To elucidate this mechanism, effects of sterol-mediated induction on both the enzyme activity of ACAT and its mRNA levels were studied in human hepatoblastoma cell line, HepG2 cells. When HepG2 cells were loaded with cholesterol and 25-hydroxycholesterol, both the whole-cell ACAT activity and the microsomal ACAT activity were increased by 85.1% and 41.3%. In contrast, cholesterol depletion of HepG2 cells with compactin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, resulted in a decrease in both the whole-cell and the microsomal ACAT activity by 46.4% and 58.3%. Under identical conditions, RT-PCR and Northern blotting analyses revealed that neither cholesterol loading nor cholesterol depletion of HepG2 cells altered the amounts of ACAT mRNA. Moreover, these treatments had no effect on the enzymatic ACAT activity determined by the reconstituted assay in which HepG2 cell homogenate had been supplemented in vitro with a saturating level of exogenous cholesterol. These results indicate that cholesterol-induced up-regulation of ACAT activity in HepG2 cells does not occur at the level of transcription, but rather at a posttranscriptional level.

Base Sequence↗
PubMed1996

Structure around C6-C7 bond of the chromophore in bathorhodopsin: low-temperature spectroscopy of 6s-cis-locked bicyclic rhodopsin analogs.

To elucidate the structural changes near the beta-ionone ring region of the chromophore during the photobleaching process of rhodopsin, the photochemical and subsequent thermal reactions of rhodopsin analogs, whose retinylidene chromophores were fixed in a 6s-cis form with a five-membered ring (6,5-rhodopsin) and a seven-membered ring (6,7-rhodopsin), respectively, were investigated by low-temperature spectroscopy. Like rhodopsin, both the rhodopsin analogs convert to the respective batho-intermediates upon absorption of light at -190 degrees C. The extinction coefficient of batho-intermediate of 6,5-rhodopsin is similar to that of bathorhodopsin, while that of 6,7-rhodopsin is considerably smaller than that of bathorhodopsin. Like bathorhodopsin, the batho-intermediate of 6,5-rhodopsin directly converts to lumi-intermediate, while that of 6,7-rhodopsin first converts to a blue-shifted intermediate and then to lumi-intermediate. These results strongly suggest that the structure around the beta-ionone ring region of the bathorhodopsin chromophore resembles 6,5-retinal rather than 6,7-retinal. From the comparison of the structural features among retinal, 6,5-retinal, and 6,7-retinal, a possible conformation around C6-C7 bond of the bathorhodopsin chromophore is discussed.

Animals↗
PubMed1996

Characterization of single chain urokinase-type plasminogen activator with a novel amino-acid substitution in the kringle structure.

ECV304 is a cell line established by a spontaneous transformation of endothelial cells of a human umbilical vein. It was shown that ECV304 secretes single chain urokinase-type plasminogen activator (scu-PA). A subclone, ECV304 clone 15, was obtained by acclimatization of parental clone to serum-free medium followed by limiting dilution. The clone was found to produce approximately five times as much scu-PA (approximately 20 IU/10(6) cells per day) as the parental clone after a 40 days' culture. Though the biochemical characteristics of the purified scu-PA were indistinguishable from those of the native scu-PA, it had a lower affinity for fibrin clots under the employed conditions. Molecular cloning of a cDNA encoding the scu-PA has identified a novel substitution from C to T in the nucleotide sequence encoding the kringle structure. The substitution resulted in an alteration from Pro (CCG) to Leu (CTG) at amino-acid position 121, which may be directly or indirectly involved in the decrease in the apparent affinity.

Amino Acid Sequence↗
PubMed1996

Cloning of the cDNA encoding the mouse ATBF1 transcription factor.

We have isolated a mouse ATBF1 cDNA which is 12-kb long and capable of encoding a 406-kDa protein containing four homeodomains and 23 zinc-finger motifs. Mouse ATBF1 is 94% homologous to the human ATBF1-A transcription factor. Northern blot and RNase protection analysis showed that levels of ATBF1 transcripts were low in adult mouse tissues, but high in developing brain, consistent with a role for ATBF1 in neuronal differentiation.

Amino Acid Sequence↗
PubMed1996

Reconstituted high density lipoprotein reduces the capacity of oxidatively modified low density lipoprotein to accumulate cholesteryl esters in mouse peritoneal macrophages.

Oxidized low density lipoprotein (ox-LDL) was incubated with discoidal complexes of apolipoprotein A-I (apo A-I) and dimyristoylphosphatidylcholine (DMPC) (DMPC/apo A-I) in a cell-free system and re-isolated on Sephacryl S-400 gel filtration chromatography. Analyses of re-isolated ox-LDL showed that apo A-I was transferred from DMPC/apo A-I to ox-LDL, which accounted for 10% of the total protein of ox-LDL. Re-isolated ox-LDL also showed a 2.2-fold increase in phospholipid and a 14% decrease in cholesterol content on an apo B basis. The electrophoretic mobility of re-isolated ox-LDL was markedly reduced almost to that of native LDL. Moreover, the amounts of re-isolated ox-LDL to be degraded by mouse peritoneal macrophages as well as the capacity of re-isolated ox-LDL to accumulate cholesteryl esters (CE) in these cells were markedly reduced (60% and 80% reduction, respectively), suggesting that the ligand activity of ox-LDL for the scavenger receptor was significantly reduced upon treatment with DMPC/apo A-I. Parallel incubation of ox-LDL with free apo A-I led to a similar incorporation of apo A-I into ox-LDL. However, it had no effects on the ligand activity of ox-LDL. Thus, it is likely that the reduction in the ligand activity of ox-LDL by DMPC/apo A-I is explained by the change in the lipid moiety (mainly phospholipid) of ox-LDL. Since discoidal high density lipoprotein (HDL) is known to occur in vivo, this phenomenon might explain one of the anti-atherogenic functions of HDL.

Animals↗
PubMed1996

Correlation between macroscopic morphologic features and malignant potential of colorectal sessile adenomas.

PURPOSE AND METHODS: To clarify malignant potential of colorectal sessile adenomas, we investigated 46 colorectal sessile adenomas surgically resected from 44 patients. Lesions were divided into three types according to their macroscopic morphologic features: nodular-type (19 adenomas), villous-type (7), and creeping-type (20). Clinicopathologic features were investigated and compared among these three types. RESULTS: Frequency of focal carcinoma in the nodular-type, villous-type, and creeping-type adenoma was 37, 29, and 55 percent, respectively. Frequency of submucosal invasion in these three types were 11, 14, and 20 percent, respectively. Irrespective of the type, large lesions (> 3 cm) tended to have a higher frequency of submucosal invasion than did smaller ones (60 vs. 20 percent; P = 0.063). Large creeping-type adenomas (> 3 cm) had a definitely higher frequency of submucosal invasion than smaller ones (36 vs. 0 percent; P < 0.05). Lymphatic or vascular invasion of carcinoma was present only in the lesion with submucosal invasion: 5 percent in the nodular-type; 14 percent in the villous-type; 20 percent in the creeping-type. Lymph node metastasis was present only in the creeping-type lesion. CONCLUSIONS: The large creeping-type adenomas (> 3 cm) have a definitely high frequency of submucosal invasive carcinoma. Endoscopic polypectomy or local resection may be adequate for most sessile adenomas. However, we recommend segmental resection of the colon with regional lymph node dissection for a creeping-type adenoma that is larger than 3 cm because of increased risk of submucosal carcinomatous invasion and lymph node involvement.

Adenoma↗
PubMed1996

Discrimination performance and resolution capacity of uncrossed visual pathways in one-eyed albino rats.

Our recent study has demonstrated that adult rats with one eye removed at birth (OEB) relearn a discrimination between alternating black and white stripes, 30 mm wide each, oriented horizontally and vertically faster than control rats monocularly enucleated in adulthood (OET), when relearning is carried out after lesions of the visual cortex and transections of the optic tract contralateral to the remaining eye. Yet, we could not obtain the corresponding results in the acquisition of the discrimination following the same surgical treatments: OEBs did master the discrimination, whereas OETs did not. We hypothesized that a large discrepancy in OETs' performance between the acquisition and relearning occurred because the stripes were close to the limit of the resolution capacity of the uncrossed visual pathways (UXVPs), and hence that a better performance of OEBs was to indicate an increase in the resolution capacity, which resulted from reorganization of the UXVPs brought about by monocular enucleation at birth. To test the hypothesis we tried to approximate the limit of the resolution capacity of the UXVPs in OEBs and OETs using seven different sized test stripes ranging from 20 to 5 mm in width after both OEBs and OETs had relearned the discrimination of the 30 mm training stripes following the same surgical treatments mentioned above. It was found that the median width of the smallest stripes for OETs to discriminate was 10 mm, and that of OEBs 7.5 mm. Although OETs could not discriminate the smallest stripes which OEBs could, they were able to discriminate stripes one third smaller than those hypothesized. Based on these findings the possibility was discussed that the acquisition as well as the relearning of the discrimination of the 30 mm stripes mediated by the UXVPs in OEBs and OETs might not be influenced by the resolution capacity, but mostly, if not entirely, by the size of the visual field.

Animals↗
PubMed1996

Differential expression of c-fos mRNA in rat prefrontal cortex, striatum, N. accumbens and lateral septum after typical and atypical antipsychotics: an in situ hybridization study.

The regional difference in the expression of c-fos mRNA induced by typical and atypical antipsychotics was determined in prefrontal cortex, striatum, N. accumbens and lateral septum in rats by in situ hybridization. Two typical antipsychotics, haloperidol (2 mg/kg) and fluphenazine (2 mg/kg), and three atypical antipsychotics, (-)sulpiride (100 mg/kg), clozapine (20 mg/kg) and OPC-14597 (40 mg/kg), were used. Brains were fixed with 4% paraformaldehyde 45 min after drug administration (i.p.). Brain sections of 30 microns-thickness were made in a cryostat and hybridized with 35S-labelled for c-fos oligonucleotide probe. These sections were apposed to X-ray films and the autoradiograms were semi-quantitatively analysed by computer-assisted densitometry. All antipsychotics used increased c-fos mRNA expression in N. accumbens shell, a region of the forebrain associated with limbic systems. On the other hand, two typical antipsychotics (haloperidol and fluphenazine) that cause a high incidence of acute motor side effects increased the expression of c-fos mRNA in the dorsolateral striatum, an extrapyramidal region primarily involved in motor control. Only clozapine induced c-fos mRNA in the medial prefrontal cortex and lateral septum. These results strongly suggest that the shell region of N. accumbens may be a common site of therapeutic action of antipsychotics.

Animals↗
PubMed1996

Scavenging effects of tea catechins and their derivatives on 1,1-diphenyl-2-picrylhydrazyl radical.

The scavenging effects of tea catechins and their epimerized, acylated, and glucostylated derivatives on 1,1-diphenyl-2-picrythydrazyl (DPPH) radical were evaluated by electron spin resonance spectrometry. Tea catechins and their epimers were shown to have 50% radical scavenging ability in the concentration range of 1 to 3 microM. No significant differences were observed between the scavenging activities of tea catechins and their epimers, and, hence, the scavenging effects of catechins are not dependent on their sterical structure. The relationship between scavenging ability and the structure of tea catechins was also examined with acylated and glucosylated catechin derivatives. It is suggested that the galloyl moiety attached to flavan-3-ol at 3 position has a strong scavenging ability on the DPPH radical as well as the ortho-trihydroxyl group in the B ring, which elevates the radical scavenging efficiency above that of the ortho-dihydroxyl group; as has been recognized in other flavonoids such as flavones. The results obtained from the reactivity of tea catechins with the DPPH radical at different pHs suggest not only that the ortho-trihydroxyl group and the galloyl moiety contribute to maintaining the DPPH radical scavenging ability more effectively in a wide range of conditions from acidic to alkaline, but also that the radical scavenging efficiency of the ortho-dihydroxyls in the B ring is limited in neutral to alkaline regions. The difference between the scavenging abilities of the trihydroxyls (probably in the galloyl moiety) and the dihydroxyls can be explained in terms of redox potentials. It is concluded that the ortho-trihydroxyl group in the B ring and the galloyl moiety at 3 position of flavan-3-ol skeleton are the most important structural features for displaying an excellent scavenging ability on the DPPH radical.

Bepridil↗
PubMed1996

A possible involvement of Fas-Fas ligand signaling in the pathogenesis of murine autoimmune gastritis.

BACKGROUND & AIMS: A Th1 clone, II-6, established from an autoimmune gastritis BALB/c mouse that underwent thymectomy 3 days after birth, recognized a 15 mer peptide constructing the alpha subunit of H+, K(+)-adenosine triphosphatase as antigen and induced gastritis in nu/nu mice by adoptive transfer. The aim of this study was to examine the molecular mechanism of target (parietal cells) destruction in either thymectomized or II-6 cell-transferred nu/nu mice. METHODS: Expression of Fas, major histocompatibility complex class II, and intercellular adhesion molecule 1 molecules on the gastric mucosa of these mice were immunohistochemically examined. In situ DNA fragmentation in these thymectomized or nu/nu mice was tested by the terminal deoxynucleotidyl transferase-mediated digoxigenin-deoxyuridine triphosphate nick end label (TUNEL) method. Moreover, activity of II-6 cells to induce apoptosis was tested by using the 15 mer peptide-pulsed B lymphoma cells, A20.2J, as the target. RESULTS: A portion of parietal cells in gastritis-bearing thymectomized or nu/nu mice at an early stage expressed Fas, major histocompatibility complex class II, and intercellular adhesion molecule 1 molecules and was TUNEL positive. Fas-ligand message was induced on activated II-6 cells and caused DNA fragmentation of the antigen-pulsed A20.2J cells. CONCLUSIONS: Cognate interaction between Fas antigen on the target and Fas ligand on the effector seems to be one possible mechanism for the target cell destruction in organ-specific autoimmune gastritis.

Animals↗
PubMed1996

Endosonographic characterization of duodenal elevated lesions.

BACKGROUND: Endoscopic diagnosis of duodenal elevated lesions is problematic for two reasons. Endoscopic biopsy often fails to confirm a histologic diagnosis of submucosal lesions. Moreover, a biopsy specimen is often insufficient to verify a differential diagnosis of mucosal lesions. In this study, we evaluated the usefulness of endosonography in the resolution of these problems. METHODS: The endoscopic and endosonographic features of 15 duodenal elevated lesions that had been confirmed histologically in our hospital were reviewed retrospectively. RESULTS: Of the 15 cases, 8 were submucosal lesions (lipoma, Brunner's gland hyperplasia, lymphangioma, carcinoid tumors, leiomyoma, and malignant lymphoma); the rest were mucosal lesions. A correct histologic diagnosis based on endoscopic biopsies was obtained in only 6 cases (three submucosal lesions and three mucosal lesions). On the other hand, ultrasonography was useful in the characterization of all submucosal lesions based on their echo level, layer of origin, and tissue homogeneity. As for mucosal lesions, the depth of infiltration was correctly estimated with endosonography. Either endoscopic resection or surgery was selected on the basis of endosonographic information. CONCLUSIONS: We conclude that endosonography is useful in the differential diagnosis of submucosal lesions and in determining suitable treatment methods for duodenal mucosal and submucosal lesions.

Biopsy↗
PubMed1996

Endoscopic resection of gastrointestinal submucosal lesions: a comparison between strip biopsy and aspiration lumpectomy.

BACKGROUND: Endoscopic diagnosis and treatment of gastrointestinal submucosal lesions is still not established. We evaluated the clinical usefulness of two resection methods for submucosal lesions, using a "nonrandomized surgeon" design. METHODS: The strip biopsy method was evaluated at Tenri Hospital and the aspiration lumpectomy method was used at Kyoto University Hospital. The inclusion criteria for selecting patients were endosonographic findings indicating a tumor location within the submucosa. RESULTS: Seventy-seven patients were treated. The size of the specimens (mean +/- SEM) was 20.7 +/- 0.9 mm for the aspiration lumpectomy group and 14.0 +/- 0.8 mm for the strip biopsy group (p < 0.01). Aspiration lumpectomy was adequate for a definitive histologic diagnosis in 95% of the cases (36 of 38) as compared with 77% (30 to 39) of the cases treated by strip biopsy (p = 0.047). The eradication rate was 87% for the aspiration lumpectomy group, and 74% for the strip biopsy group (p = 0.274). The complication rate resulting from these procedures (hemorrhage, mean 5%) was acceptable. Repeated endoscopic examinations revealed no recurrence except for one lymphangioma. CONCLUSIONS: Endoscopic resection, especially aspiration lumpectomy, provides a significant benefit for accurate final diagnosis and eradication of submucosal lesions.

Adolescent↗
PubMed1996

Developmental changes in expression of the ATBF1 transcription factor gene.

The expression of the ATBF1 gene in developing brain was analyzed in mice from 13 days of gestation to 28 days after birth using RNase protection and in situ hybridization methods. The level of ATBF1 transcripts was the highest on embryonic day 13-15 and then progressively decreased to a hardly detectable level on postnatal day 28. Throughout the period examined, ATBF1 mRNA was expressed consistently in the basal telencephalon, diencephalon, and mesencephalon, with the highest levels in the inferior colliculus and thalamus. On the other hand, no significant expression was observed in cerebellum, neocortex, hippocampus, and olfactory bulb. These results illustrate significant regional differences in the ATBF1 expression and suggest a role of the ATBF1 gene in the formation of some specific cell populations in developing central nervous system.

Animals↗
PubMed1996

Immunological and virological study of sudden deafness.

Thirty-three patients with sudden deafness and 11 controls were selected from the patients admitted to the Department of Otolaryngology, Tokai University Hospital from November 1990 to October 1991. Viral titers were measured for mumps, adenovirus, rubella, measles, herpes simplex virus (HSV), varicella zoster virus (VZV), rhinosyncytial virus, cytomegalo-virus (CMV), and mycoplasma pneumoniae in 33 sudden deafness patients and 11 controls at a 2-week interval. In 20 of 33 sudden deafness patients and 5 of 11 controls, autoantibodies of rheumatoid factor (RF), anti-mitochondrial antibody (AMA), anti-nuclear antibody (ANA), anti-parietal cell antibody (APA), anti-smooth muscle antibody (ASA), and anti-type II collagen antibody were studied. Viral titer study did not reveal any significant change either in the patients or in the controls, whereas autoantibody study revealed a relatively high incidence for ASA in the patients as compared with the controls. The relatively high incidence for ASA suggests that immune-mediated processes may be involved in the etiology of sudden deafness.

Adenoviridae↗
PubMed1996

Crystal structure of the macrophage migration inhibitory factor from rat liver.

The tertiary structure of the macrophage migration inhibitory factor (MIF) from rat liver (12,300 Mr) is presented at 2.2 A resolution. Each monomer consists of two beta/alpha/beta motifs aligned in quasi two-fold symmetry, comprising a domain consisting of a four-stranded mixed beta-sheet and two antiparallel alpha-helices. The protein exists as a trimer in the crystal. An extra beta-strand that is almost perpendicular to the other beta-strands joins to the beta-sheet of the neighbouring monomer in the trimer. Unexpected similarities were detected between MIF and two kinds of isomerase.

Amino Acid Sequence↗