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Biomedical subjects

T Harada

Publications and source records attributed to T Harada.

At least 541 records · Page 30Linked to original sources
PubMed1993

An efficient expression vector for stable expression in human liver cells.

The expression of the bacterial neomycin resistance (NmR)-encoding gene was examined under control of the promoter region of the gene encoding the human polypeptide chain elongation factor 1 alpha (EF-1 alpha). In a human liver cell line, HepG2, the plasmid pEF321-neo directed higher expression of the bacterial neo gene than the NmR gene expression vectors using other promoters, like the SV40 early region or thymidine kinase of Herpes simplex virus and SV40 early region promoter linked to human T-cell leukemia virus-1 enhancer sequences.

Cell Line↗
PubMed1993

Possible pathogenic role of cationic anti-DNA autoantibodies in the development of nephritis in patients with systemic lupus erythematosus.

Extensive studies in murine models of lupus nephritis have shown that cationic anti-DNA autoantibodies have nephritogenic potential. We have investigated whether cationic anti-DNA antibodies of IgG class are also produced in vivo in patients with active lupus nephritis. Antibodies against DNA in the sera from patients with SLE were purified by affinity chromatography on DNA-cellulose, followed by nonequilibrium pH gradient electrophoresis and SDS-PAGE. The highly cationic anti-DNA antibodies of IgG class were prominent in the nonequilibrium pH gradient electrophoresis-immunoblots of the antibodies from the patients with active lupus nephritis. Decreased proteinuria after successful treatment with prednisolone was associated with disappearance of the cationic anti-DNA antibodies in the circulation. The cationic anti-DNA antibodies did bind to heparan sulfate, which is a major glycosaminoglycan in glomerular basement membrane, much better than did neutral anti-DNA antibodies. The results suggest that the cationic anti-DNA autoantibodies may play a certain role in the development of lupus nephritis. Our study demonstrates the strong association between the presence of cationic anti-DNA antibody and the development of lupus nephritis in humans.

Adolescent↗
PubMed1993

Feedback regulation mechanisms for the control of GTP cyclohydrolase I activity.

Guanosine triphosphate (GTP) cyclohydrolase I, the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin (BH4), is subject to feedback inhibition by BH4, a cofactor for phenylalanine hydroxylase. Inhibition was found to depend specifically on BH4 and the presence of another protein (p35). The inhibition occurred through BH4-dependent complex formation between p35 protein and GTP cyclohydrolase I. Furthermore, the inhibition was specifically reversed by phenylalanine, and, in conjunction with p35, phenylalanine reduced the cooperativity of GTP cyclohydrolase I. These findings also provide a molecular basis for high plasma BH4 concentrations observed in patients with hyperphenylalaninemia caused by phenylalanine hydroxylase deficiency.

Animals↗
PubMed1993

Relationship between osteopenia and clinical characteristics of Parkinson's disease.

Pathological bone changes affect locomotor activity and may influence the outcome and prognosis of Parkinson's disease (PD). In this study, we determined the relationship between bone changes and PD. Bone study was performed by multiple scanning x-ray photodensitometry (MD/MS) in 64 patients with PD and 42 age-matched controls. We then compared the results with the clinical characteristics of PD. Osteopenia was detected in 22 (53.6%) of the 41 female and 6 (26%) of the 23 male patients, and in 6 (26%) of the 23 female and 2 (10.5%) of the 19 male controls. The frequency of osteopenia was significantly greater in the female patients than in the male patients or the female controls. Osteopenia was related to the duration of PD in the men, but not in the women. Twenty of 40 PD patients' hands showed side-related differences in the analysis of both hands. In 19 of the 20 patients, the side of more severe osteopenia coincided with that of parkinsonian symptoms, suggesting that osteopenia is related to the pathophysiology of PD.

Absorptiometry, Photon↗
PubMed1993

The effect of spermine on the disaccharidase activities in suckling rats of different age.

The influence of the age of the rat on the maturation of disaccharidase activities induced by spermine was studied. Three-day- and 9-day-old rats were used in the experiment. Spermine was administered orally or directly into the stomach using thin tubing daily for 3 days, and disaccharidase activities in the jejunum were measured. While spermine caused maturation of not only lactase activity but also maltase and sucrase activities in 9-day-old rats, it only caused maturation of lactase activity in 3-day-old rats. Histological studies showed no significant changes in the jejunum of 3-day-old rats treated with spermine.

Age Factors↗
PubMed1993

Quantitative analysis of neuronal damage induced by tri-ortho-cresyl phosphate in Wistar rats.

A quantitative analysis of neuronal damage was performed on the fasciculus gracilis (FG) of the cervical spinal cord in male Wistar rats that received orally a single dose of tri-ortho-cresyl phosphate (TOCP) at 1500 mg/kg. FG tissues were sampled at 1, 2, and 3 weeks after treatment and examined histopathologically. Wallerian degeneration of myelinated nerve fibers was observed in FG at 2 weeks. Morphological changes were most evident at 3 weeks after treatment and the number of fibers was reduced. Ultrastructurally, axonal swelling due to the accumulation of cytoplasmic contents was observed near the node of Ranvier in the affected animals, indicating paranodal degeneration. Axonal atrophy and swelling in organophosphorus-induced delayed neuropathy (OPIDN) were evaluated quantitatively using a computer-assisted image analyzer. Morphometric examinations on semi-thin sections and frozen sections stained with Nauta's method were demonstrated to be useful for objective evaluation of OPIDN in the rat.

Animals↗
PubMed1993

Arachidonic acid metabolism by isolated and cultured middle ear epithelial cells from the chinchilla.

Thin-layer chromatography was used to examine the metabolism of arachidonic acid and prostaglandins (PGs) in freshly isolated and cultured middle ear epithelial cells from the chinchilla. The freshly isolated cells converted arachidonic acid predominantly to PGE2, while those cells grown in culture for 10 days acquired the ability to convert arachidonic acid to 6-keto-PGF1 alpha, PGD2, and PGE2. Incubation of the isolated cells and primary cultures with acetylsalicylic acid and indomethacin inhibited the formation of these PGs. These findings suggest that studies on the factors regulating arachidonic acid metabolism in middle ear epithelium may help to explain the role of eicosanoids in middle ear secretions, particularly in relation to the pathophysiology of otitis media.

6-Ketoprostaglandin F1 alpha↗
PubMed1993

Biochemical characterization of mucous glycoproteins secreted by in vitro chinchilla middle ear epithelial cells.

A method for middle ear epithelial (CMEE) cell culture with active mucus secretory function has been successfully developed, using the chinchilla as an animal model. CMEE cells were dissociated by protease digestion from the middle ear mucosa. The CMEE cells grown in primary culture incorporated [3H]glucosamine into a glycoconjugate after its release into medium. This substance was characterized biochemically as mucin, although the production of mucin by the cells required growth on a substratum of collagen gel. These cultures provide an excellent model for studying factors that regulate synthesis and secretion of glycoproteins in CMEE cells.

Animals↗
PubMed1993

An electrocochleographic study of acute low-tone sensorineural hearing loss.

Twenty-four patients with acute low-tone sensorineural hearing loss (ALHL) were examined using electrocochleography. The negative summating potential (SP) amplitude and the summating potential/action potential (AP) ratio were significantly greater in the ALHL patients than in normals. The SP/AP ratio was smaller in the ALHL patients than in patients with known Meniere's disease and moderate hearing loss, although the SP amplitude was somewhat greater in the former. An abnormal increase in the SP amplitude following click stimuli was found in 54% of the ALHL patients, while the SP/AP ratio was increased abnormally in 63% of these patients. These findings suggest that the pathophysiology of ALHL may be similar to that for endolymphatic hydrops.

Acoustic Stimulation↗
PubMed1993

Relationship between immunological parameters and survival of patients with liver metastases from breast cancer given immuno-chemotherapy.

We treated 33 patients with liver metastases from breast cancer by immuno-chemotherapy including adoptive cell transfer between 1987 and 1992. In this study, we examined the change of immunological parameters in the peripheral blood lymphocytes and interleukin-2 (IL-2)-cultured lymphocytes, in primary vs. metastatic breast cancer patients and before vs. after treatment. Moreover, we examined their correlation with therapeutic response and survival after treatment. The immunological parameters used were in vitro natural killer cell activity (% lysis of K562), in vitro autologous tumor-killing activity (% lysis against autologous freshly isolated tumor cells), and proliferation of lymphocytes stimulated with IL-2 and autologous sonicated tumor extract antigen in mixed culture (IL-2-enhanced MLTR). When compared with primary breast cancer patients, patients with liver metastases showed a significant decrease in % lysis of K562 and autologous tumor cells. After treatment, the stimulation index in IL-2-enhanced MLTR increased significantly from the pretreatment level and correlated with survival after treatment. Moreover, non-specific immunological parameters (performance status, lymphocyte count, and transferred cell count and proliferation rate of cultured lymphocytes) were significantly associated with response and prognosis.

Adult↗
PubMed1993

Does critical swimming velocity represent exercise intensity at maximal lactate steady state?

The purpose of this investigation was to determine whether the critical swimming velocity (vcrit), which is employed in competitive swimming, corresponds to the exercise intensity at maximal lactate steady state. vcrit is defined as the swimming velocity which could theoretically be maintained forever without exhaustion and expression as the slope of a regression line between swimming distances covered and the corresponding times. A total of eight swimmers were instructed to swim two different distances (200 m and 400 m) at maximal effort and the time taken to swim each distance was measured. In the present study, vcrit is calculated as the slope of the line connecting the two times required to swim 200 m and 400 m. vcrit determined by this new simple method was correlated significantly with swimming velocity at 4 mmol.l-1 of blood lactate concentration (r = 0.914, P < 0.01) and mean velocity in the 400 m freestyle (r = 0.977, P < 0.01). In the maximal lactate steady-state test, the subjects were instructed to swim 1600 m (4 x 400 m) freestyle at three constant velocities (98%, 100% and 102% of vcrit). At 100% vcrit blood lactate concentration showed a steady-state level of approximately 3.2 mmol.l-1 from the first to the third stage and at 98% of vcrit lactate concentration had a tendency to decrease significantly at the fourth stage. On the other hand, at 102% of vcrit, blood lactate concentration increased progressively and those of the third and fourth stages were significantly higher than those at 100% of vcrit (P < 0.05). These data suggest that vcrit, which can be calculated by performing two timed, maximal effort swimming tests, may correspond to the exercise intensity at maximal lactate steady state.

Adolescent↗
PubMed1993

The therapeutic effect of OK-432-combined adoptive immunotherapy against liver metastases from gastric or colorectal cancers.

Twenty-four patients with liver metastases from gastric or colorectal cancer were treated with OK-432-combined adoptive immunotherapy (AIT). Lymphocytes isolated from regional lymph nodes or peripheral blood were cultured with medium containing T cell growth factor and sonicated tumor extract antigen (SE-Ag) for 9-13 days. The cultured lymphocytes were transferred mainly through the hepatic artery after the administration of OK-432, a streptococcal preparation. Sixteen of the 24 patients received a low dose of anti-cancer agents between the OK-432 injection and cell transfer. When cultured without SE-Ag, regional lymph node lymphocytes (RLNL) showed significantly (P < 0.05) higher cytotoxic activity against autologous tumor cells and, on the contrary, lower cytotoxic activity against K562 than peripheral blood lymphocytes (PBL). When cultured with SE-Ag, cytotoxicity of RLNL against autologous tumor cells was nearly equivalent to that of PBL. The blastogenesis of fresh PBL to SE-Ag was significantly (P < 0.05) augmented after the OK-432-combined AIT. Two patients showed complete response and 4 patients showed partial response among 19 patients who had evaluable lesions. Five patients whose liver metastases were resected were treated with OK-432-combined AIT as an adjuvant therapy. To date they are alive without recurrence in the liver.

Adult↗
PubMed1993

Cellular interaction against autologous tumor cells between IL-2-cultured lymphocytes and fresh peripheral blood lymphocytes in patients with breast cancer given immuno-chemotherapy.

In patients with Stage II or III breast cancer and in patients with liver metastases from breast cancer, we examined cellular interaction in the cytotoxicity against autologous tumor cells by interleukin-2(IL-2)-cultured lymphocytes (CL) and fresh peripheral blood lymphocytes (FPBL) treated with immunochemotherapy including OK-432 and cyclophosphamide. In flow cytometric analysis, CD8+CD11b+ and CD16+ cells significantly decreased after immuno-chemotherapy in both groups of patients. A protocol study in Stage II or III breast cancer patients showed suppressive activity of FPBL on the cytotoxic activity of CL in 3/9 of the non-treatment group but no suppressive activity and enhancing activity in 3/7 in the immuno-chemotherapy group. Moreover, in 19 patients with liver metastases from breast cancer treated with immuno-chemotherapy including adoptive immunotherapy, FPBL in 6/19 showed enhancing activity, and in 8/19 suppressive activity in the lysis of autologous tumor cells. In assays in vitro using autologous and allogeneic tumor cells, FPBL showed a partial specificity in cellular interaction against autologous tumor cells. CD4-depleted FPBL inhibited cytotoxicity of CL, while CD8-depleted FPBL enhanced cytotoxicity of CL in patients with liver metastases. These results suggest that immuno-chemotherapy eliminates the suppressive population in FPBL and may induce tumor regression if combined with adoptive immunotherapy using CL.

Adjuvants, Immunologic↗
PubMed1993

The induction of murine tumor infiltrating lymphocytes (TIL) by interleukin-2 or T cell growth factor.

Mice were injected in the foot pad with either 5 x 10(5) syngeneic plasmacytoma (MOPC104E) or fibrosarcoma cells (Meth A). Lymph nodes containing tumor cells were harvested 14 days later and cultured. In the presence of recombinant interleukin-2 (r-IL-2) predominantly tumor cells proliferated. Culture with T cell growth factor (TCGF) resulted in the growth of lymphoid cells. Concanavalin A (Con A) had only a modest effect on elimination of tumor cells in the culture. Tumor-infiltrating lymphocytes (TIL) prepared from the lymph nodes showed specific tumor-neutralizing activity when grown in the presence of TCGF. In vitro examination revealed that Meth A cells could not be lysed by TIL, while TIL from MOPC tumors showed tumor specific activity. This study may explain negative results in human trials with TIL induced by IL-2 alone.

Animals↗
PubMed1993

Relationship between biliary excretion of bilirubin and glutathione disulfide.

The effects of two glutathione-oxidizing agents, t-butyl hydroperoxide and diamide, on biliary excretion of bilirubin and glutathione disulfide were investigated in anesthetized male Sprague-Dawley rats. Bilirubin (unconjugated) was infused at a constant rate of 100 nmol/kg/min through the jugular vein. When biliary excretion of bilirubin was stabilized, either of the glutathione-oxidizing agents was administered via the mesenteric vein. Biliary excretion of glutathione disulfide increased temporarily after the administration and returned to its basal levels within 20 min. The biliary excretion of bilirubin decreased during the same period and returned to the former levels thereafter. Changes in bile flow rates remained within 20% of the basal levels. A linear correlation was found between the increments in the bile concentration of glutathione disulfide and the decrements in that of bilirubin. Furthermore, separate experiments revealed that reduction of hepatocellular glutathione per se had little effect on biliary excretion of bilirubin. The results thus indicate that the reduction of biliary excretion of bilirubin by glutathione-oxidizing agents was due to the increase in biliary excretion of glutathione disulfide, and suggest that a common biliary excretory mechanism is shared, at least partially, by bilirubin and glutathione disulfide in Sprague-Dawley rats.

Animals↗
PubMed1993

Antioxidant protection against oxidant-induced damage in cultured gastric mucosal cells.

Gastric epithelium is exposed not only to oxidants generated within the lumen, but also to those produced by ischemia/reperfusion. This study examined the mechanism(s) of oxidant-induced injury to cultured rat gastric mucosal cells, and characterized the antioxidant profile of these cells. Hydrogen peroxide (H2O2), generated by glucose oxidase, damaged cells dose-dependently, as assessed by increased leakage of labeled 51Cr. Glucose oxidase-induced damage was prevented by exogenous catalase (but not by exogenous superoxide dismutase). Chelation of cellular iron with desferrioxamine or phenanthroline specifically protected cells against H2O2, whereas binding of extracellular iron with apotransferrin failed to. Disruption of the glutathione redox cycle at three independent sites rendered cells less resistant to H2O2, whereas inhibition of cellular catalase did not result in sensitization of cells to H2O2. In conclusion, (1) oxidant injury induced by extracellular H2O2 is mediated by intracellular iron; (2) extracellular superoxide is not involved in the damaging process; and (3) the glutathione redox cycle plays a principal role in detoxifying H2O2 as a cellular antioxidant in cultured gastric mucosal cells.

Animals↗