Search PubMed⌕ Search

Biomedical subjects

K Saeki

Publications and source records attributed to K Saeki.

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

Arsenic accumulation in three species of sea turtles.

Arsenic in the liver, kidney and muscle of three species of sea turtles, e.g., green turtles (Chelonia mydas), loggerhead turtles (Caretta caretta) and hawksbill turtles (Eretmochelys imbricata), were determined using HG-AAS, followed by arsenic speciation analysis using HPLC-ICP-MS. The order of arsenic concentration in tissues was muscle > kidney > liver. Unexpectedly, the arsenic concentrations in the hawksbill turtles feeding mainly on sponges were higher than the two other turtles primarily eating algae and mollusk which accumulate a large amount of arsenic. Especially, the muscles of the hawksbill turtles contained remarkably high arsenic concentrations averaging 153 mg kg(-1) dry weight with the range of 23.1-205 mg kg(-1) (n = 4), even in comparison with the data from other organisms. The arsenic concentrations in the tissues of the green turtles were significantly decreased with standard carapace length as an indicator of growth. In arsenic compounds, arsenobetaine was mostly detected in the tissues of all the turtles. Besides arsenobetaine, a small amount of dimethylarsinic acid was also observed in the hawksbill turtles.

Animals↗
PubMed2000

Bcl-2 down-regulation causes autophagy in a caspase-independent manner in human leukemic HL60 cells.

To understand the roles of bcl-2 for the survival of leukemic cells, we constructed human leukemic HL60 transformant lines in which full length bcl-2 antisense message was conditionally expressed by a tetracycline-regulatable expression system. Cell growth was completely inhibited after antisense message induction and massive cell death was induced. Electron microscopic examinations show that cells died by autophagy, but not by apoptosis. The morphology and the function of mitochondria remained intact: neither the reduction in mitochondrial membrane potential nor the nuclear translocation of AIF, a mitochondrial protein that translocates to nuclei in cases of apoptosis, was observed. Caspase inhibitors did not rescue bcl-2-antisense-mediated autophagy. Thus, bcl-2 is essential for leukemic cell survival and its down-regulation results in autophagy. Cell Death and Differentiation (2000) 7, 1263 - 1269.

Anti-Bacterial Agents↗
PubMed2000

Induction of apoptosis in human hematopoietic U937 cells by granulocyte-macrophage colony-stimulating factor: possible existence of caspase 3-like pathway.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) induced apoptosis in human hematopoietic U937 cells by itself and in a synergistic manner with tumor necrosis factor (TNF). GM-CSF-induced apoptosis was not inhibited by caspase inhibitors YVAD-CMK, DEVD-CHO and z-VAD-FMK, under the condition that these inhibitors potently suppressed TNF-induced apoptosis. Both GM-CSF and TNF induced caspase 3-like activity in this cell line though the time course was distinct between two cytokines, and combined stimulation of cells with GM-CSF plus TNF induced additive or synergistic activation of caspase 3-like activity. Amount of immunoreactive cleaved forms of caspase 3 recognized by specific antibody was completely dissociated with its enzymatic activity when the cells were stimulated with GM-CSF, but not with TNF. These results indicate that GM-CSF induces apoptosis of U937 cells via unknown pathway, which seems to be mediated by caspase 3-like activity, yet not caspase 3 itself, resistant to the caspase inhibitors, and synergistically interacts with conventional caspase 3 pathway of TNF. Possible involvement of caspases 1 and 8 (-like activity) but not caspase 7 in this pathway was also suggested.

Apoptosis↗
PubMed2000

Neuronal and vascular localization of histamine N-methyltransferase in the bovine central nervous system.

Histamine N-methyltransferase (HMT) (EC 2.1.1.8) plays a crucial role in the inactivation of the neurotransmitter histamine in the CNS. However, the localization of HMT remains to be determined. In the present study, we investigated immunohistochemical localization of HMT in the bovine CNS using a polyclonal antibody against bovine HMT. The HMT-like immunoreactivity was observed mainly in neurons. Strongly immunoreactive neurons were present in the oculomotor nucleus and ruber nucleus in the midbrain, the facial nucleus in the pons, the dorsal vagal nucleus and hypoglossal nucleus in the medulla oblongata and in the anterior horn as well as intermediolateral zone of the spinal cord. Intermediately immunoreactive neurons were present in the piriform cortex and the inferior olivary nucleus. The grey matter of the forebrain regions was diffusely and faintly stained. In the cerebellum and the striatum, the nerve fibres in the white matter were positive. The tuberomammillary nucleus, where histaminergic neurons are present, were weakly positive. The other immunoreactive structures in the CNS were blood vessels. Almost all of the blood vessel walls, irrespective of whether they were arterial or venous, were variably stained. The glial fibrillary acidic protein- (GFAP-) immunoreactive astrocytes were not stained. These findings indicated that histamine released from histaminergic nerve terminals or varicose fibres is methylated mainly in postsynaptic or extrasynaptic neurons rather than in astrocytes. The localization of HMT in the blood vessel wall may mean that blood-borne histamine and histamine released from mast cells associated with the blood vessels are catabolized in this structure.

Animals↗
PubMed2000

Purification and characterization of a trypsin-like serine proteinase from rat brain slices that degrades laminin and type IV collagen and stimulates protease-activated receptor-2.

A trypsin-like serine proteinase was purified from the incubation medium of rat brain slices by gelatin zymography. The purification consisted of ammonium sulfate precipitation, benzamidine-Sepharose 6B affinity chromatography, and carboxymethyl-cellulose and gel filtration chromatographies. The gelatinolytic activity, identified at 22 kDa (P22) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under nonreducing conditions, was eluted as one active peak throughout the purification, and the final preparation gave a single protein peak on reverse-phase HPLC. Diisopropyl fluorophosphate, benzamidine, p-toluenesulfonyl-L-lysine chloromethyl ketone, and aprotinin completely inhibited the activity of P22, whereas phenanthroline, p-toluene-sulfonyl-L-phenylalanine chloromethyl ketone, and elastinal did not. P22 efficiently digested the extracellular matrix proteins laminin and type IV collagen. P22 produced an increase in intracellular Ca2+ concentration in A172 glioblastoma, which was desensitized through prior stimulation with protease-activated receptor-2 agonist peptide SLIGKV, indicating that P22 can stimulate protease-activated receptor-2. Rat brain penetration injury induced gelatinolytic activity in the lesioned area whose molecular size was consistent with that of P22. These results indicated that on incubation of rat brain slices, a trypsin-like serine proteinase was secreted into the medium that was capable of digesting extracellular matrix and stimulating protease-activated receptor-2. It is suggested that the gelatinolytic activity induced by brain injury might be that of P22.

Animals↗
PubMed2000

Apoptosis-inducing activity of galloyl monosaccharides in human histiocytic lymphoma U937 cells.

Three galloyl monosaccharides contained in medicinal plants were examined for apoptosis-inducing activity in human histiocytic lymphoma U937 cells. Tetragalloyl glucose (TgG) induced apoptosis as found by chromatin condensation, DNA ladder formation, and inhibition by a caspase inhibitor. Digalloyl hamamelose had moderate activity, while monogalloyl glucose was only marginally active. These findings suggest that the number and disposition of their phenolic groups are important for apoptosis induction. TgG induced apoptosis in human colon and stomach cancer cell lines as well, indicating it is potentially useful as an anti-cancer agent.

Antineoplastic Agents, Phytogenic↗
PubMed2000

Improvement in development of porcine embryos reconstituted with cells from blastocyst-derived cell lines and enucleated oocytes by optimization of reconstruction methods.

The present study was conducted to establish the most suitable system for producing porcine reconstructed embryos by transferring cells from blastocyst-derived cell lines into enucleated oocytes. When the cells were fused to preactivated metaphase II oocytes, or the cells and arrested metaphase II oocytes were fused in medium without CaCl(2) and MgSO(4), the percentages (43-53%) of fused embryos were significantly lower than those (72-79%) produced by fusing the cells to arrested metaphase II oocytes in medium containing CaCl(2) and MgSO(4). High productive efficiency (7%) of blastocysts was obtained when reconstituted embryos produced by the last method were activated again at 3 hours after fusion (F/A --> Activation). Pronuclear formation was observed in 80-91% of the reconstructed embryos produced by F/A --> Activation, with no significant differences between different culture periods in the medium containing cytochalasin B. When cultured in the medium containing cytochalasin B for 0-1 h, almost all (83-85%) the embryos had one pronucleus and one polar body. However, the number of embryos with two pronuclei and no polar bodies was increased significantly by culturing in the medium containing cytochalasin B for 2-4 h. The cleavage rate (34-48%) of reconstructed embryos was not affected by the presence of cytochalasin B for 2 h after activation. However, the percentage of embryos that developed to the blastocyst stage was significantly higher in the presence (23%) than absence (5%) of cytochalasin B. The results indicate that F/A --> Activation and cytochalasin B treatment are effective for the production of porcine embryos reconstituted with cells from blastocyst-derived cell lines and enucleated oocytes.

Animals↗
PubMed2000

Significant overexpression of metallothionein and cyclin D1 and apoptosis in the early process of rat urinary bladder carcinogenesis induced by treatment with N-butyl-N-(4-hydroxybutyl)nitrosamine or sodium L-ascorbate.

Effects of a genotoxic bladder carcinogen, N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) and a non-genotoxic bladder promoter, sodium L-ascorbate (Na-AsA), on protein expression, cell proliferation and apoptosis of the bladder epithelium with or without the influence of testicular castration were investigated. Male F344 rats were divided into six groups (groups 1-6). BBN was given with 0.05% drinking water to groups 1 and 4 for 8 weeks, groups 2 and 5 received diet with 5% Na-AsA. Then the animals were treated without any chemicals. Groups 3 and 6 were non-treated controls. Testicular castration was carried out 2 weeks before commencement of chemical treatment on groups 4-6. The total observation period was 18 weeks. Overexpression of cyclin D1 was induced by BBN but not Na-AsA and the degree of overexpression was higher in the order simple hyperplasia, papillary or nodular hyperplasia, papilloma and carcinoma. Metallothionein (MT) was also overexpressed in bladder epithelium treated with BBN but not Na-AsA, but was decreased in papillomas and never found in a carcinoma. Cyclin D1-positive cells were essentially MT-negative. Therefore, it is speculated that MT protects genes from insult by genotoxic carcinogens and its lack is associated with tumor development. Apoptotic cell death occurred during treatment with BBN and Na-AsA and after their withdrawal. Chromatin condensation of many G0/G(1) cells was particularly marked on flow cytometry analysis 1 week after cessation of treatment, this being considered as an early apoptotic change. Although testicular castration had no influence on the above events, it resulted in decreased tumor formation as compared with the case of similarly treated intact animals. Our data demonstrate that overexpression of MT and cyclin D1 is specific for treatment with a genotoxic carcinogen, and suggest that MT overexpression may play an important suppressive role in the early stages of rat urinary bladder carcinogenesis.

Animals↗
PubMed2000

Functional medical ligament balancing in total knee arthroplasty.

Function of the anterior and posterior oblique portions of the medial collateral ligament and the posterior capsule in flexion and extension was evaluated in eight knee specimens after posterior cruciate retaining total knee arthroplasty. The posterior oblique portion of the medial collateral ligament was released subperiosteally in four specimens, and the anterior portion was released in four specimens. The medial posterior capsule was released in each group, then the remaining portion of the medial collateral ligament was released. Release of the posterior oblique portion produced moderate laxity at full extension and at 30 degrees flexion, and posterior capsule release produced additional laxity in full extension. Release of the anterior portion produced major laxity at 60 degrees and 90 degrees flexion. Complete medial collateral ligament release increased laxity significantly in both groups in flexion and extension. This rationale was tested in a clinical study of 82 knees (76 patients) in which 62 (76%) required medial collateral ligament release to correct varus deformity during posterior cruciate retaining total knee arthroplasty. Twenty-two knees (35.5%) were tight medially in extension only, and were corrected by releasing the posterior oblique portion. Thirty-one knees (50%) were tight medially in flexion only, and were corrected by releasing the anterior portion. Nine knees (14.5%) were tight medially in flexion and extension and required complete medial collateral ligament release, but three knees (4.8%) remained tight in extension and required medial posterior capsule release to correct flexion contracture and medial ligament contracture. Seventeen (27%) had partial posterior cruciate ligament release to correct excessive rollback of the femoral component on the tibial surface.

Arthroplasty, Replacement, Knee↗
PubMed2000

Immunohistochemical localization of histamine N-methyltransferase in guinea pig tissues.

Histamine plays important roles in gastric acid secretion, inflammation, and allergic response. Histamine N-methyltransferase (HMT; EC 2.1.1.8) is crucial to the inactivation of histamine in tissues. In this study we investigated the immunohistochemical localization of this enzyme in guinea pig tissues using a rabbit polyclonal antibody against bovine HMT. The specificity of the antibody for guinea pig HMT was confirmed by Western blotting and the lack of any staining using antiserum preabsorbed with purified HMT. There was strong HMT-like immunoreactivity (HMT-LI) in the epithelial cells in the gastrointestinal tract, especially in the gastric body, duodenum, and jejunum. The columnar epithelium in the gallbladder was also strongly positive. Almost all the myenteric plexus from the stomach to the colon was stained whereas the submucous plexus was not. Other strongly immunoreactive cells included the ciliated cells in the trachea and the transitional epithelium of the bladder. Intermediately immunoreactive cells included islets of Langerhans, epidermal cells of the skin, alveolar cells in the lung, urinary tubules in the kidney, and epithelium of semiferous tubules. HMT-LI was present in specific structures in the guinea pig tissues. The widespread distribution of HMT-LI suggests that histamine has several roles in different tissues.

Animals↗
PubMed2000

Apoptosis-inducing activity of lipid derivatives of gallic acid.

Gallic acid derivatives with a lipophilic group (hydrogenated farnesyl gallate, lauryl gallate, gallic acid laurylamide and cholesteryl gallate) were examined for their ability to induce apoptosis in human monoblastic leukemia U937 cells. Farnesyl ester derivative is the most potent apoptosis inducer among the compounds examined. The results suggest that lipid derivatives can augment the apoptosis-inducing activity of gallic acid depending on the structure. These findings will provide useful information in developing anti-cancer agents.

Apoptosis↗
PubMed2000

Clastogenicity of quinoline derivatives tested by micronucleus induction in vivo in the hepatocytes of partially hepatectomized mice.

The induction of micronucleated liver cells (MN-liver cell) was examined with halogenated and hydroxylated quinolines using partially hepatectomized mice. Among the chloroquinolines, 8-chloroquinoline demonstrated a significantly higher level of induction than the control. All the fluorinated derivatives examined, except for 6-fluoroquinoline, induced significantly higher levels, and there were no appreciable differences in MN-liver cell induction among the fluorinated quinolines, regardless of their mutagenic potencies in the Ames test. Of the hydroxylated quinolines examined, 2- and 4-isomers, which are not mutagenic, induced MN-liver cells to the same extent as a mutagenic isomer, 8-hydroxyquinoline. It seems that clastogenicity was not satisfactorily correlated with mutagenicity in the Ames test as far as this class of compounds is concerned.

Animals↗
PubMed2000

[Anti-carcinogenic structural modification by fluorine-substitution in aza-polycyclic aromatic hydrocarbons].

One of the major goals of a series of our studies is to explore the availability of a method for anti-genotoxic modification of carcinogens by fluorine-substitution. Quinoline, a hepatocarcinogen, mutates bacterial tester strains in the presence of rat liver microsomal enzymes and induces GST-P (placental glutathione S-transferase)-positive foci in a medium-term bioassay system for hepatocarcinogenesis. On the other hand, 3-fluorinated quinoline (3-FQ) was neither mutagenic nor carcinogenic in the same assay system, whereas 5-fluoroquinoline (5-FQ) was mutagenic and carcinogenic. Quinoline, 3-FQ, and 5-FQ were also tested in an in vivo mutagenicity assay system using a lacZ-transgenic mouse (Muta Mouse). Mutation was induced by quinoline and 5-FQ only in the liver, the target organ of carcinogenesis by quinoline, but not in the other organs examined. 3-FQ was non-mutagenic in all of the organs. The results strongly indicate that fluorine-substitution at the position-3 of quinoline could be an anti-genotoxic structural modification of quinoline in a wide range of its genotoxic end-points. Additionally, seventeen mono- and di-fluorinated derivatives of 1,7-phenanthroline, 1,10-phenanthroline, benzo[h]quinoline, and benzo[f]quinoline were subjected to analysis of their structure-mutagenicity relationship. The results support that the enamine epoxide structure of the pyridine moiety, as well as the bay-region epoxide structure, is responsible for mutagenicity. These results suggest that the introduction of a fluorine atom to the molecule in question may be a useful tool to modify their mutagenic potency and to better understand the mechanism of mutation.

Animals↗
PubMed2000

Distance education for health centre staff in rural Japan.

We have evaluated a course delivered by videoconferencing to rural health centre staff in Hokkaido. The course focused on the planning and evaluation of community health interventions. It included four 90 min sessions and two follow-up sessions. Fourteen professional staff members (public health nurses, nutritionists and dental assistants) attended each class. Knowledge of community health-care planning and evaluation was higher at post-education testing than pre-education testing. Ratings for 'using a computer', 'using some computer software', 'using the Internet' and 'interest in telehealth' increased significantly in post-education testing compared with pre-education testing. The course had an additional benefit in increasing the collaboration between community health workers and university staff.

Adult↗
PubMed2000

Upgrading rural Japanese nurses' respiratory rehabilitation skills through videoconferencing.

We examined the effect of distance learning on nurses' clinical skills in a rural Japanese hospital. The subject matter was respiratory rehabilitation. After one face-to-face session, who 30 min sessions were delivered by videoconferencing to staff nurses working in a 100-bed rural hospital 250 miles (400 km) from Sapporo Medical University. A self-rating questionnaire was distributed before and after the sessions. Responses were collected from 15 out of 32 nurses participating in the face-to-face session (47%). Before the first videoconferencing session, 'always use' and 'sometime use' the new skills were rated by 67% of nurses, but after the second videoconferencing session 'always use' and 'sometimes use' were rated by 73% and 'never use' at 0%. This implies that there was increased use of new skills after the second session, although the difference was not significant. The nurses' opinions about the effectiveness for patients increased from 8% to 27% after the second session, which was significant. The pilot project indicated the usefulness of distance learning for upgrading nurses' clinical practice in one rural Japanese hospital and suggested ways in which videoconferencing can be used in future.

Attitude of Health Personnel↗
PubMed2000

Japanese scrapie cases.

Worldwide attention has been given to scrapie, because bovine spongiform encephalopathy (BSE) could be experimentally transmitted to sheep. This ovine form of BSE was clinically identical to scrapie. In Japanese scrapie cases, a majority of the diseased sheep were from Suffolk, while 8 cases were from Corriedale. It is very likely that sheep-to-sheep transmission of scrapie has taken place in Obihiro, Hokkaido. Normal prion protein may play a role in the morphoregulatory signaling pathway, which orchestrates the specificity of a particular cellular response. Over-expression of normal prion protein in mice cause neurodegenerative disorders. Recently, Prnd was identified downstream of the mouse prion protein gene (Prnp), and encodes 179 amino acids and a prion protein (PrP)-like protein designated doppel (Dpl). Dpl was upregulated in the central nervous system of two PrP-deficient lines of mice, as well as in prionless cell lines. Dpl caused neurodegeneration similar to that caused by PrP. Linked expression of Prnp and Prnd may cause several neurodegenerative disorders.

Animals↗
PubMed1999

Aberrant expression of cAMP-response-element-binding protein ('CREB') induces apoptosis.

We have reported previously that cAMP-response-element-binding protein (CREB) was phosphorylated in a cell-cycle-dependent manner, showing that it was phosphorylated at early S-phase at casein kinase II target sites. To assess the possible involvement of CREB in cell cycle progression, CREB expression vector was transiently transfected into various cells. Unexpectedly we found that transfection with CREB expression vector resulted in an abundance of dead cells. Morphological examination revealed that these cells had undergone apoptosis. The coincidence of CREB overexpression and apoptosis induction at the individual cell level was confirmed by a immunohistochemical study. To confirm that overexpression of CREB was the cause of apoptosis, a dominant-negative mutant of CREB, KCREB, was co-expressed with the wild type. The co-existence of KCREB effectively rescued CREB-mediated apoptosis in a dose-dependent manner, verifying that apoptosis was truly a specific effect of overexpressed CREB and not an artifact of the transfection procedure. Deletion analysis indicates that neither the Q1 transactivation domain, which functions in transcription, nor the kinase-inducible domain, in which a cluster of various kinase targets exists, is necessary; however, the Q2 transactivation domain is required for the induction of apoptosis. A more precise study indicates that the four-residue stretch Glu-Glu-Ala-Ala at the most C-terminal region of the Q2 domain is especially important for the induction of apoptosis. Thus overexpressed CREB induces apoptosis by transmitting certain signals from the C-terminal portion of the Q2 domain. Possible roles of cell-cycle-regulated phosphorylation and also an elevation of the intracellular cAMP level in CREB-induced apoptosis are suggested.

Amino Acid Sequence↗