Biomedical subjects

K Haraguchi

Publications and source records attributed to K Haraguchi.

At least 73 records · Page 4Linked to original sources

PubMed1998

Catalepsy induced by calcium channel blockers in mice.

It is known that calcium channel blockers induce Parkinsonism. In this study, amlodipine-, diltiazem-, and verapamil-induced catalepsy was investigated in mice. All of these three calcium channel blockers induced catalepsy. Dopamine D1, D2, and mACh receptor occupancies were estimated under the same conditions, and the affinities of these drugs for each receptor were also estimated in vitro. Intensity of catalepsy was predicted by dopamine D1, D2, and mACh receptor occupancies with the dynamic model which had already been constructed and was compared with the observed values. The predicted and the observed values were comparable (r = 0.98, p < 0.001). In conclusion, the dynamic model considering D1, and D2, and mACh receptor occupancy may be useful for quantitative prediction of drug-induced catalepsy.

Amlodipine↗

PubMed1998

Inhibition of cell-cell communication by methylsulfonyl metabolites of polychlorinated biphenyl congeners in rat liver epithelial IAR 20 cells.

The effects of three polychlorinated biphenyl (PCB) congeners and their six methylsulfonyl (MeSO2)-metabolites on cell communication have been investigated in the scrape-loading/dye-transfer assay in IAR 20 rat liver epithelial cells. The results demonstrated that at non-cytotoxic concentrations 2,2',4',5-tetrachlorobiphenyl, 2,2',4',5,5'-pentachlorobiphenyl (2,2',4',5,5'-pentaCB), 2,2',4',5,5',6-hexachlorobiphenyl (2,2',4',5,5', 6-hexaCB), and their 3- and 4-MeSO2 derivatives completely inhibited the cell communication within 1 h. 4-MeSO2-2,2',4',5,5'-pentaCB and 4-MeSO2-2,2',4',5, 5',6-hexaCB appeared to inhibit the cell communication at slightly lower concentration than their parental PCB congeners and 3-MeSO2 derivatives. The results show that 3- and 4-MeSO2 derivatives of the PCB congeners tested inhibit gap junction intercellular communication at about the same potency as their parental compounds. Since inhibition of cell communication is often observed after treatment with many tumor promoters, our findings suggest that the metabolites may also act as tumor promoters.

Alkanesulfonates↗

PubMed1998

Methyl sulphone metabolites of polychlorinated biphenyls (PCBs) in cetaceans from the Irish and the Aegean Seas.

The levels of PCBs and PCB methyl sulphone metabolites were determined in the blubber of six species of cetaceans that originated from the Irish Sea and the Aegean Sea (Mediterranean). Burdens ranging from 2.80 to 27.80 microg g-1 (lipid weight) total PCBs and 0.03 to 0.58 microg g-1 (lipid weight) total methyl sulphones were recorded. The highest cetacean methyl sulphone burden (0.58 microg g-1) was in an Irish Sea harbor porpoise (Phocoena phocoena). Methyl sulphone isomer formation capacity, as determined by PCB:MSF ratios, varied considerably between species as follows: harbour porpoise (1:10) > pilot whale (Globicephalus melas), white-sided dolphin (Lagenorhyncus acutus) (1:50) > common dolphin (Delphinus delphis), Risso's dolphin (Grampus griseus), and striped dolphin (Stenella coerueoalba), (1:100), and was related to interspecific variation in PCB methyl sulphone formation capacity.

Adipose Tissue↗

PubMed1998

Hydroxylation and methylthiolation of mono-ortho-substituted polychlorinated biphenyls in rats: identification of metabolites with tissue affinity.

The metabolism of three mono-ortho-substituted congeners, 2,3,3',4, 4'-pentachlorobiphenyl (CB105), 2,3',4,4',5-pentachlorobiphenyl (CB118), and 2,3,3',4,4',5-hexachlorobiphenyl (CB156), was investigated with regard to the identification of hydroxy- and sulfur-containing metabolites and their tissue retention in rats. Hydroxylation proceeded primarily at the meta or para position either via an arene oxide, involving NIH shift and dechlorination, or by direct insertion of a hydroxyl group. CB105 was hydroxylated preferably in the 2,3,4-trichlorinated ring to yield 4-OH-2,3,3',4', 5-pentaCB, whereas CB118 was hydroxylated in the 2,4, 5-trichlorinated ring to yield the same hydroxy metabolite to a similar extent. The concentration of 4-OH-2,3,3',4',5-pentaCB in blood was >3 times higher than that in liver, lung, or kidney. The ratios of 4-OH-2,3,3',4',5-pentaCB to unchanged CB in blood were 11:1 for CB105 and 7:1 for CB118. The other two metabolites, 4'-OH-2, 3',4,5,5'-pentaCB from CB118 and 4'-OH-2,3,3',4,5,5'-hexaCB from CB156, also exhibited a high blood affinity. Another metabolism of mono-ortho-PCBs PCBs involved methylthiolation in the vicinal ortho and meta unsubstituted positions to give methylthio metabolites, which were detected as methylsulfonyl metabolites in liver and adipose tissue. The tissue retention of these metabolites might contribute to the toxic and biologic effects of mono-ortho-substituted PCBs.

Animals↗

PubMed1998

Effects of cytokines on expression of thyrotropin receptor mRNA in rat preadipocytes.

Cultured rat preadipocytes express thyrotropin receptor (TSHR) during their differentiation. To evaluate the effects of inflammatory cytokines on the expression of TSHR in cultured rat preadipocytes, we cultured those cells in the presence of recombinant human tumor necrosis factor (rhTNF)-alpha, recombinant human interferon (rhIFN)-gamma, and human transforming growth factor (hTGF)-beta1. The effects on the level of TSHR mRNA and signal transduction were evaluated. Addition to the medium of 1 ng/mL TNF-alpha, 1 ng/mL rhIFN-gamma, and 1 ng/mL hTGF-beta1 during the differentiation of rat preadipocytes inhibited the expression of TSHR mRNA. The decrease in TSHR mRNA was accompanied by a decrease in TSH-stimulated cyclic adenosine monophosphate (cAMP) production. Histochemical analysis showed that these cytokines inhibited the morphological differentiation of the cells. These cytokines also decreased the expression of mRNA for such fat-specific proteins as lipoprotein lipase and aP2. Results indicate that the loss of expression and function of the TSHR is closely related to the inhibition of differentiation. This confirms the close relation between the expression of the TSHR and the differentiation of the rat preadipocytes.

Adaptor Protein Complex 2↗

PubMed1998

The glycosylphosphatidylinositol-anchored phosphatase from Spirodela oligorrhiza is a purple acid phosphatase.

We recently presented clear evidence that the major low-phosphate-inducible phosphatase of the duckweed Spirodela oligorrhiza is a glycosylphosphatidylinositol (GPI)-anchored protein, and, to our knowledge, is the first described from higher plants (N. Morita, H. Nakazato, H. Okuyama, Y. Kim, G.A. Thompson, Jr. [1996] Biochim Biophys Acta 1290: 53-62). In this report the purified 57-kD phosphatase is shown to be a purple metalloenzyme containing Fe and Mn atoms and having an absorption maximum at 556 nm. The phosphatase activity was only slightly inhibited by tartrate, as expected for a purple acid phosphatase (PAP). Furthermore, the protein cross-reacted with an anti-Arabidopsis PAP antibody on immunoblots. The N-terminal amino acid sequence of the phosphatase was very similar to those of Arabidopsis, red kidney bean (Phaseolus vulgaris), and soybean (Glycine max) PAP. Extracts of S. oligorrhiza plants incubated with the GPI-specific precursor [3H]ethanolamine were treated with antibodies raised against the purified S. oligorrhiza phosphatase. Radioactivity from the resulting immunoprecipitates was specifically associated with a 57-kD band on sodium dodecyl sulfate-polyacrylamide gels. These results, together with previous findings, strongly indicate that the GPI-anchored phosphatase of S. oligorrhiza is a PAP.

Acid Phosphatase↗

PubMed1998

Analysis of differentiation-induced expression mechanisms of thyrotropin receptor gene in adipocytes.

Rat adipose tissue, as well as differentiated 3T3-L1 cells, has been shown to express TSH receptor (TSHR) mRNA in amounts approaching those in the thyroid. We investigated the molecular mechanisms of TSHR gene expression in adipose cells. Primer extension and cloned cDNA sequences showed that transcription of the TSHR gene in rat adipose tissue was from multiple start sites clustered between -89 to -68 bp and almost identical to those in FRTL-5 thyroid cells. By transient expression analysis, we localized, between -146 and -90 bp, a positive regulatory element, the activity of which was markedly increased after the differentiation of 3T3-L1 cells. Deoxyribonuclease I protection showed that nuclear extracts from differentiated 3T3-L1 cells strongly protected two sequences, from -146 to -127 bp, including a cAMP response element-like sequence and from -112 to -106 bp containing a putative Ets-binding sequence. In differentiated 3T3-L1 cells, disruption or deletion of either sequence was found to result in the loss of enhancer activity, suggesting both elements may synergistically activate the TSHR promoter. Electrophoretic mobility shift analysis revealed the induction of new protein/DNA complexes formed either with the cAMP response element-like site or with putative Ets elements after the differentiation into adipocytes. In contrast, nuclear proteins, whose binding to DNA was diminished after the differentiation of 3T3-L1 cells, were found to interact with the site contiguous to the 5'-end of the putative Ets-binding sequence. Mutations of this binding site, which reduced the protein/DNA complex formation, increased TSHR promoter activity in undifferentiated cells. These observations suggested that differentiation-induced diminution of suppressor interactions may allow the enhancers to synergistically activate the transcription of TSHR gene in adipocytes.

Adipocytes↗

PubMed1998

Transient isolated thyrotropin deficiency associated with cavernous sinus syndrome.

The cause and course of isolated thyrotropin (TSH) deficiency are not well understood. We report a 65-year-old man with a transient, probable isolated TSH deficiency associated with cavernous sinus syndrome secondary to tympanitis. On his admission, serum TSH and triiodothyronine levels were very low. No TSH response to thyrotropin releasing hormone (TRH) was observed. However, 6 years later, TSH response to TRH was restored. The present case showed that inflammation in the cavernous sinus could be one of the causes of TSH deficiency. Further, it demonstrated that TSH deficiency is not always permanent and the reevaluation of pituitary function is necessary.

Aged↗

PubMed1998

[Effective splenectomy in a patient with HIV-associated thrombocytopenia].

A 31-year-old man presented with a 3-month history of petechial hemorrhages. Physical examination revealed no splenomegaly. The patient's platelet count was 1.0 x 10(9)/l and bone marrow aspiration showed an elevated number of megakaryocytes. A diagnosis of HIV-associated thrombocytopenia was made on the basis of HIV seropositive results. The CD4 cell count was 400 x 10(6)/l. No opportunistic infections indicating AIDS were detected. Initially the patient was treated with predonisolone, but showed only a transient response. He also failed to respond to zidovudine, lamivudine, or indinavir. Following splenectomy, however, his platelet count rose above 80 x 10(9)/l (normal level: 150-350 x 10(9)/l).

Adult↗

PubMed1998

[The Osaka concept. "Dome osteotomy" with of without labrum resection].

Acetabular dysplasia represents the most common aetiology for secondary osteoarthritis of the hip joint in Japan. On radiographs progress of the disease can be classified in four stages (I to IV). Labral lesions are common accompanying findings in acetabular dysplasia, representing an important prognostic factor. Therefore we routinely use conventional arthrography to classify labral lesions in three types (normal, torn and detached). From 1978 to 1983 a modified Chiari osteotomy of the pelvis (dome osteotomy) was performed in 64 hip joints without arthrotomy. The preoperative radiography showed labral lesions in 66%. After an average follow-up of 4 years, 43 of the 44 patients with a normal or torn labrum showed excellent or good results. On contrast, 50% of the 20 patients with a detached labrum showed fair and poor results only. From 1984 to 1989 an arthrotomy was combined with dome osteotomy in 29 hip joints and labral lesions had to be resected in 16 cases. The results were generally good in patients with arthrosis grade I and II, but in stage III they were only fair. Thus, the success of labral surgery can only be achieved in early stages of dysplasia. Based on our findings dome osteotomy combined with arthrotomy and labral surgery can be recommended in patients with labral lesions and hip dysplasia grade I and II.

Acetabulum↗

PubMed1997

Activation of serum response factor in the liver of Long-Evans Cinnamon (LEC) rat.

We have studied the DNA binding activities of transcription factors in the liver of Long-Evans Cinnamon (LEC) rats, an animal model of Wilson's disease. Owing to a genetic defect, this strain of rats accumulates excessive copper in the liver and develops severe hepatitis and hepatocellular carcinoma. We found that the DNA binding activity of the serum response factor (SRF) was higher in the liver of LEC rats (approximately 2-fold) than in that of Wistar rats. There was a close correlation between the intensity of the activity and the concentrations of copper in the nuclear protein. The DNA binding activity of Sp1, on the other hand, showed similar levels in both LEC and Wistar rats. SRF may play an important role in the development of hepatocellular carcinoma in LEC rats by mediating the proto-oncogene c-fos induction. We suggest that the copper in nuclear protein may be involved in the activation of SRF.

Animals↗

PubMed1997

Socioeconomic factors and dental caries in developing countries: a cross-national study.

The purpose of the present study was to analyze statistically correlations between socioeconomic factors and the prevalence of dental caries in developing countries. The DMFT index, which evaluates the incidence of dental caries, showed a positive correlation (P < 0.01) with several socioeconomic factors, such as life expectancy, adult literacy rate, school attendance rate, population employed in the service sector, population aged 15-64 years, and urban population. According to multiple regression analysis, population aged 15-64 years, population employed in the service sector, and urban population were the most influential independent socioeconomic variables, in descending order, with a regression coefficient of 0.635 and a coefficient of determination of 0.404 (P < 0.001). This finding suggests that the prevalence of dental caries in developing countries increases with the degree of urbanization.

Adolescent↗

PubMed1997

Metabolism of 3,3',4,4'-tetrachlorobiphenyl via sulphur-containing pathway in rat: liver-specific retention of methylsulphonyl metabolite.

1. Single doses of 3,3',4,4'-tetrachlorobiphenyl (TCB) were administered intraperitoneally to the male Wistar rat for metabolism studies. 2. Seven sulphur-containing metabolites of TCB were isolated from faeces, in addition to previously reported 4-hydroxy-3,3',4',5-tetrachlorobiphenyl and 5-hydroxy-TCB. Major sulphur-containing metabolites were 5- and 6-methylthio-TCBs, and 6-methylsulphonyl-3-methylthio-3',4,4'-trichlorobiphenyl. 3. The faecal excretions of hydroxy metabolites, methylthio metabolites and unchanged TCB accounted for 7.1, 0.5 and 0.3% of the dose respectively within 4 days after administration. 4. 5-Methylsulphonyl-TCB was detected and selectively retained in liver. The concentration ratio of 5-methylsulphonyl-TCB and unchanged TCB in liver was 1:4. 5. Following administration of 5- and 6-methylsulphonyl-TCBs to rat, 5-methylsulphonyl-TCB was localized in liver, whereas 6-methylsulphonyl-TCB was rapidly biotransformed to 6-methylsulphonyl-3-methylthio-3',4,4'-trichlorobiphenyl and excreted in the faeces.

Adipose Tissue↗

PubMed1997

Epstein-Barr virus-positive multiple early gastric cancers and dysplastic lesions: a case report.

Epstein-Barr virus (EBV) has been implicated as a causal virus of gastric cancer with episomal monoclonality, elevated antibodies and a unique morphologic expression in the early intramucosal stage, but the infection mechanisms have not been demonstrated. EBV has been shown only in the cancerous lesions by the highly sensitive EBV-encoded small RNA in situ hybridization (EBER-ISH) method, not in the dysplastic mucosa adjacent to the cancer. A case is presented of multiple EBV-positive gastric cancer and dysplastic epithelium observed in a 52-year-old man. Serial cut sections of the gastrectomy specimen showed four small cancerous lesions, three of which were EBER-positive, and three EBER-positive, minute, non-cancerous dysplastic lesions. The three cancerous lesions were intramucosal cancer, with one having minimal submucosal invasion forming a lymphoepithelioma-like histology. All of these EBER-positive cancerous and dysplastic lesions showed intense CD8 T-lymphocytic infiltration. There was no such findings in the EBV-negative cancerous lesion. It was concluded that EBV infection may occur in the epithelial cells of atrophic gastric mucosa, and progress to cancer with monoclonal expansion through the EBV-positive dysplastic change. Cytotoxic T-lymphocytic reactions can occur even in the dysplastic lesions. Multifocal EBV infection in the gastric mucosa may occur and, if necessary, total gastrectomy is recommended in such a case.

Herpesviridae Infections↗

PubMed1997

Iodide uptake and experimental 131I therapy in transplanted undifferentiated thyroid cancer cells expressing the Na+/I- symporter gene.

131I therapy is a widely accepted treatment for differentiated thyroid cancers which can accumulate iodide. We evaluated the efficiency of 131I therapy against tumors which are transfected with the Na+/I- symporter (NIS) gene. We transfected the rat NIS cDNA expression vector into malignantly transformed rat thyroid cells (FRTL-Tc) which do not concentrate iodide. The resultant cell line (Tc-rNIS) accumulated 125I 60-fold in vitro. The FRTL-Tc cells formed solid tumors after injection of cells into subcutaneous tissues of Fischer 344 rats. Tumors formed with Tc-rNIS cells accumulated up to 27.3% of total 125I administered, and concentrated 125I 11 to 27-fold in the tumors. Extracorporeal measurement of radioactivity in the tumors revealed that 125I accumulation peaked at 90 min, and decreased to half levels 6 h after the injections. To investigate the effect of 131I administration on the tumor growth, we injected Na131I 2 and 3 weeks after the transplantation of the cells. The Na131I did not change the tumor volume significantly in either the FRTL-Tc or the Tc-rNIS-induced tumors. The short (6 h) effective half life of 131I in the tumors diminished the radiation dose to the tumor cells. However, this approach may prove beneficial in the treatment of radiosensitive cancers, and could be employed diagnostically.

Animals↗

PubMed1997

Regulation of thyrotropin receptor gene expression in 3T3-L1 adipose cells is distinct from its regulation in FRTL-5 thyroid cells.

We previously have demonstrated that rat adipose tissue expresses TSH receptor (TSHR) messenger RNAs (mRNAs) at levels approaching those detected in the thyroid. Furthermore, we recently reported that TSHR mRNA is detected in fibroblast-like 3T3-L1 cells after their hormone-induced differentiation into adipocytes. TSH induces cAMP formation and lipolysis in differentiated 3T3-L1 cells. We now show that, in Northern blot analyses, TSH-induced down-regulation of TSHR mRNA levels, which can be duplicated by forskolin and dibutylyl cAMP, i.e. which is cAMP-mediated. We also have demonstrated that a beta-adrenergic stimulant, which stimulates cAMP formation in adipocytes, induces a down-regulation of TSHR mRNA levels in 3T3-L1 adipocytes. Nuclear run-on assays show that the ability of TSH/cAMP to decrease TSHR mRNA levels in 3T3-L1 cells reflects transcriptional regulation. This report also demonstrates that TSHR gene expression in 3T3-L1 adipocytes is regulated in a manner distinct from that observed in thyroid cells. Thus, in fully differentiated 3T3-L1 adipocytes, TSH-induced down-regulation of TSHR mRNA levels is evident within 1 h and is near maximum within 4 h after addition of TSH. A transient increase of TSHR gene expression, which has been demonstrated in FRTL-5 thyroid cells, was not observed in 3T3-L1 adipocytes. The down-regulation of TSHR gene expression induced by TSH/cAMP in 3T3-L1 cells is cycloheximide-insensitive, suggesting that continuous protein synthesis is not required for this process. In contrast, the down-regulation of TSHR gene expression observed in FRTL-5 cells is sensitive to cycloheximide. In both FRTL-5 thyroid cells and 3T3-L1 adipocytes, insulin or serum increased TSHR mRNA levels. Although insulin or serum was required for the TSH-induced down-regulation of TSHR mRNA levels in FRTL-5 thyroid cells, neither insulin nor serum was required for TSHR down-regulation in 3T3-L1 adipocytes. These findings demonstrate that TSH/cAMP regulates TSHR mRNA levels in adipocytes via a regulatory system distinct from that used in FRTL-5 cells. This report further demonstrates that adipose cells do not express thyroid transcription factor-1, which interacts with the TSHR promoter region in FRTL-5 cells, and that 3T3-L1 nuclear extracts exhibit a different binding activity to the cAMP-response element-like element in the TSHR promoter region compared with extracts from FRTL-5 cells.

3T3 Cells↗

PubMed1997

Purification and characterization of the acid soluble 26-kDa polypeptide from soybean seeds.

Whey proteins from soybean seeds of Japanese varieties were analyzed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Among 11 varieties of soybean, three green and one black soybeans lacked a 26-kDa band that was found in all yellow soybeans. In this paper, the 26-kDa protein was named AS26k (acid soluble 26-kDa protein) temporarily. The AS26k protein was purified from Glycine max cv. Nattosyoryu, which is yellow soybean, through four purification steps: 30-35% saturated ammonium sulfate fractionation, ion exchange chromatography on S Sepharose Fast Flow, gel filtration on Sephadex G-100, and hydrophobic chromatography on phenyl Sepharose CL-4B. Purified AS26k was cleaved with V8 proteinase from Staphylococcus aureus or CNBr. The cleaved polypeptide contained two typical dehydrin motif sequences: DEYGNPV and (M)DKIKEKLPG, and a 19 amino acids sequence similar to a pea dehydrin. Native AS26k had a molecular mass of 32 kDa on gel filtration and a pl of 7.2 on two-dimensional PAGE. Similarly to other dehydrins and late embryogenesis abundant (LEA) proteins, AS26k was rich in hydrophilic amino acids, and highly heat stable. These results showed that AS26k was a dehydrin, a group II LEA protein in soybean seeds.

Amino Acid Sequence↗

PubMed1997

Prediction of drug-induced catalepsy based on dopamine D1, D2, and muscarinic acetylcholine receptor occupancies.

It is known that catalepsy serves as an experimental animal model of parkinsonism. In this study, the relationship between in vivo dopamine D1 and D2 receptor occupancies and catalepsy was investigated to predict the intensity of catalepsy induced by drugs that bind to D1 and D2 receptors nonselectively. 3H-SCH23390 and 3H-raclopride were used for the labeling of D1 and D2 receptors, respectively. The ternary complex model consisting of agonist or antagonist, receptor, and transducer was developed, and the dynamic parameters were determined. After coadministration of SCH23390 and nemonapride, catalepsy was stronger than sum of the values predicted by single administration of each drug, and it was intensified synergistically. This finding suggested the existence of interaction between D1 and D2 receptors, and the necessity for constructing the model including this interaction. To examine the validity of this model, catalepsy and in vivo dopamine receptor occupancy were measured after administration of drugs that induce or have a possibility to induce parkinsonism (haloperidol, flunarizine, manidipine, oxatomide, hydroxyzine, meclizine, and homochlorcycilzine). All of the tested drugs blocked both dopamine D1 and D2 receptors. Intensity of catalepsy was predicted with this dynamic model and was compared with the observed values. In contrast with haloperidol, flunarizine, manidipine, and oxatomide (which induced catalepsy), hydroxyzine, meclizine, and homochlorcyclizine failed to induce catalepsy. Intensities of catalepsy predicted with this dynamic model considering the interaction between D1 and D2 receptors overestimated the observed values, suggesting that these drugs have catalepsy-reducing properties as well. Because muscarinic acetylcholine (mACh) receptor antagonists inhibit the induction of catalepsy, the anticholinergic activities of the drugs were investigated. After SCH23390, nemonapride and scopolamine were administered simultaneously; catalepsy and in vivo mACh receptor occupancy were measured to evaluate quantitatively the anticholinergic activity. Relationship between mACh receptor occupancy and change in catalepsy was used as the measure of catalepsy-reducing effects of the drugs. Measurement of in vivo mACh receptor occupancy revealed a significant blockade of mACh receptor by all of the tested drugs except for haloperidol. The predicted values of catalepsy, when corrected for the mACh receptor-related reduction, approached the observed values. This finding indicates the possibility that mACh receptor antagonism of drugs may contribute to the reduction of catalepsy. In conclusion, the dynamic model considering D1, D2, and mACh receptor occupancies and synergism between D1 and D2 receptors may be useful for quantitative prediction of drug-induced catalepsy.

Animals↗