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Biomedical subjects

H Saeki

Publications and source records attributed to H Saeki.

At least 127 records · Page 7Linked to original sources
PubMed1994

Gene expression of protein phosphatases in rat ascites hepatoma cell lines.

Gene expression of protein phosphatases 1 alpha, 1 gamma 1, 2A alpha, and 2C alpha in 14 rat ascites hepatoma cell lines was studied by Northern blot hybridization. The expression of PP1 alpha and PP2C alpha was increased and decreased, respectively, in all of the ascites hepatoma (AH) cells compared to rat liver, whereas the expression of PP1 gamma 1 and PP2A alpha was increased in about 50% of them. Relative gene expression was affected by several factors, such as harvest time, transplantation rate, percentage of free cells, and sex; the first factor was more important than the others. Relative gene expression of PP1 alpha had a negative correlation with harvest time, whereas gene expression of PP1 gamma 1 and PP2A alpha had a nonlinear (hyperbolic) correlation with harvest time. We suggest that there is a relationship between growth rate and expression of protein phosphatase genes. Our data also suggest that PP1 gamma 1 mRNA is positively controlled by PP2A alpha mRNA.

Animals↗
PubMed1994

[Studies on hypogeusia in hyperemesis gravidarum].

We measured changes in gustatory sensitivities and gustation threshold levels and those of some trace elements, vitamin E and lipids in the serum of 38 patients suffering from hyperemesis gravidarum, 12 normal pregnant women in the first trimester and 22 healthy non-pregnant women by means of filter discs and an electrogusto meter (EGM) and we obtained the following results. 1. Gustatory sensitivity to sweetness was found to be significantly lowered in the patients with hyperemesis gravidarum when tested with filter discs (p < 0.05). 2. Gustation threshold levels were significantly lowered in the patients with hyperemesis gravidarum and the normal pregnant women tested with the EGM (p < 0.01). 3. There was considerable heterogeusia of women in the patients with hyperemesis gravidarum (p < 0.05). 4. Zn and Mg in serum decreased (p < 0.01), but Cu increased significantly in the patients with hyperemesis gravidarum and the normal pregnant women (p < 0.01). 5. Total cholesterol and phospholipid in serum significantly decreased, but lipid peroxide in serum significantly increased in the patients with hyperemesis gravidarum (p < 0.01). These results suggested that the gustatory test was helpful in investigating hyperemesis gravidarum and that biological defense systems (scavenger systems) to free radicals were damaged in patients with hyperemesis gravidarum.

Adult↗
PubMed1993

A novel human monoclonal antibody directed to a tumor-associated antigen.

Twelve human monoclonal antibodies (HuMAb) were established by the fusion of (mouse x human) heteromyeloma cells with B-lymphoblastoid cells derived from the regional lymph nodes of three patients with squamous cell carcinoma of the lung. They were tested for reactivity to two kinds of proteins (purified protein derivatives and bovine serum albumin) by ELISA, Sq-19 (squamous cell carcinoma) culture cells by indirect membrane immunofluorescence tests, and Sq-19 tumor xenograft by immunohistological study. Among them, one HuMAb 904F (IgM, lambda) was selected. In indirect membrane immunofluorescence tests, this 904F antibody reacted with various kinds of cell lines, e.g. lung cancer, esophageal cancer, endometrial cancer, and stomach cancer. It did not react with malignant hematopoietic and diploid fibroblast cell lines. Immunohistologically, it stained the tumor nests of squamous cell carcinoma, adenocarcinoma, and large cell carcinoma of the lung. It also stained those of esophagus and colon, but not those of small cell carcinoma of lung, or stomach. On frozen-section specimens of normal tissues from various organs, it showed only limited areas of positive staining. Limited positive findings were observed at a reticular zone of the adrenal gland, at the esophagus as weak staining, and at islets of the pancreas as very weak staining. Western blotting analysis demonstrated that it recognized a 54 kDa trypsin-sensitive molecule which is expressed on the surface of tumor cells. These results suggest the 904F monoclonal antibody detects a novel tumor-associated antigen which is recognized by the human immune system.

Animals↗
PubMed1993

Production of a human monoclonal antibody to a synthetic peptide by active in vivo immunization using a SCID mouse grafted with human lymphocytes.

In order to meet the present increased demands, we have tried to improve the methods to produce human monoclonal antibodies by using a SCID mouse grafted with human mononuclear cells. Initially, we gave an anti-asialo GM1 antibody to a SCID mouse to suppress the NK activity, as a pretreatment. Then, fifty million human mononuclear cells (MNC) from a healthy volunteer were injected intraperitoneally to the SCID mouse so as to construct a human immune system in the mouse (PBL-SCID mouse). We immunized the mouse with a synthetic peptide (pep 190) conjugated with KLH four times. The spleen cells taken from the immunized PBL-SCID mouse were fused with (mouse x human) heteromyeloma cells. The hybridoma cells were selected in GIT medium containing HAT and IL-6. Among 68 hybridoma-growing wells, we obtained one hybridoma clone (#41-1-4) which secreted a specific antibody to pep 190. The reactivity of this monoclonal antibody was tested by ELISA and the specificity of this antibody was confirmed by an absorption test with different kinds of proteins. This paper is the first report of the successful production of peptide-specific human monoclonal antibody by active in vivo immunization using a PBL-SCID mouse. By this active in vivo immunization system using a PBL-SCID mouse, human monoclonal antibodies for any sort of peptide antigens may easily be made available.

Amino Acid Sequence↗
PubMed1993

Prognostic significance of proliferating cell nuclear antigen (PCNA) expression in non-small cell lung cancer.

Formalin-fixed, paraffin-embedded tissue sections of resected tumors from 90 patients were immunohistochemically studied to assess the prognostic value of proliferating cell nuclear antigen (PCNA) expression in non-small cell lung cancer. The individual tumors were classified into groups of high, moderate or low proliferative grade, and 38 (42.2%) patients had a high grade of proliferation. No statistically significant correlations were observed between PCNA grade and TNM status, pathological stage, resectability, histological type, degree of histological differentiation. Only vascular invasion significantly correlated with proliferative grade (p < 0.05). Survival analysis showed that patients with low proliferative grade tumors survived significantly longer (a 5-year survival rate of 83.3%) than those with high proliferative grade tumors (39.4%, p < 0.005). Cox's multivariate analysis revealed that PCNA grade was a significant prognostic determinant of survival. These results suggest that PCNA expression provides an independent prognostic variable for patients with non-small cell lung cancer and that it may be useful to consider this factor in treatment planning.

Aged↗
PubMed1993

[Measurement and its fluctuation of urinary glucaric acid in newborns].

It is known that urinary excretion of glucaric acid(GA) is an indirect index of hepatic P-450 microenzyme induction. We measured and analyzed urinary excretion of GA in 56 newborns and 26 mothers by a new method for the inhibition of beta-glucuronidase activity and obtained the following results. 1. The concentration of urinary GA was correlated with that of urinary creatinine and total bilirubin in newborns. 2. There were no significant correlations between gestational age, sex, body weight at birth, placental weight and the urinary GA concentration. 3. The urinary excretion of GA in newborns was decreased the in first few days after birth, but a transitional increase was observed on the fifth day after birth. 4. The concentration of urinary GA was correlated with that of direct bilirubin in serum on the fifth day after birth. 5. There was a negative correlation between the urinary GA concentration on the first day after birth and that of direct bilirubin in serum on the fifth day after birth. These results suggested that hepatic P-450 microsomal enzyme was induced by bilirubin in newborns and it was possible to estimate the clinical course of jaundice by measuring the urinary excretion of GA.

Bilirubin↗
PubMed1992

Surgical repair versus medical treatment of varicocele in the rat: pharmacological manipulation of the varicocelized testicle.

Human chorionic gonadotropin, kallikrein, indomethacin, and hydralazine were administered to different groups of varicocelized rats, while surgical repair of the varicocele was performed in another group of rats. The effects of conservative and surgical treatment on epididymal sperm content and motility, the weights of the testes, epididymis, and male accessory genital glands, and fertility were compared between each group and a sham-treated group of rats. Surgical repair significantly improved all the evaluated parameters and all the conservative regimens, except hydralazine, resulted in a significant improvement in most parameters. Our results indicate that stimulation of the Leydig or/and Sertoli cells of a varicocelized testicle can counteract some of the detrimental consequences of the varicocele itself.

Animals↗
PubMed1992

New strategies to establish human monoclonal antibodies.

In order to establish human monoclonal antibodies to any sort of antigens efficiently, we have made following two approaches. Our first approach is to improve cell fusion frequency. By improving our previous method for production of human hybridomas, we obtained higher frequency (1/700 vs. 1/5500) compared with our previous method by adding irradiated myeloma cells to culture of fusion cells and modifying the selective medium. Our second approach is to use a SCID-hu mouse for immunization. Since the injection of human PBL can result in the stable long-term reconstitution of a human immune system in SCID mouse, we tried to immune SCID-hu mouse with KLH. In the serum of immunized SCID-hu mouse, we obtained human IgG antibodies to KLH. Additionally, we succeeded in establishing human B lymphoblastoid cell lines which produced antibodies specific to KLH. These methods will open new prospects for the detection and therapy of cancer.

Absorption↗
PubMed1992

[Case report of Farmer's lung disease in siblings in Kagoshima Prefecture].

Two cases of farmer's lung disease in siblings are reported. A 54-year-old male farmer, who had been engaged in stock work for 20 years, presented to our clinic for the second episode of fever, productive cough and shortness of breath. Chest roentgenogram revealed diffuse micronodular pattern, and mild hypoxemia was recognized on arterial blood gas analysis. Cytology obtained from BALF showed lymphocytosis, with especially increased OKT3, OKT4 positive cells and OKT4/8 ratio. The diagnosis was confirmed by highly positive reaction of precipitins to Thermoactinomyces vulgaris and granulomatous interstitial pneumonitis on histopathological examination. The second case was a 51-year-old female patient, the sister of the first case, who also worked as a stock farmer of another farm for 20 years. She presented with an episode of similar symptoms to the first case, one and a half years after the onset of her brother's symptoms. The findings of roentgenogram, BALF analysis, precipitins to T. Vulgaris and pathology were similar to those of the first case. The finding of high OKT4/8 ratio on BALF analysis in both cases is characteristic of formers lung disease, in contrast to the summer-type hypersensitivity pneumonitis which usually shows low OKT4/8 ratio in BALF. To our knowledge, this is the first report of farmer's lung disease in an area other than the northern part of Japan. Thus we conclude that farmer's lung disease may occur in Japan under any environmental conditions, and that some genetic factor may take part in the onset of this disease.

Bronchoalveolar Lavage Fluid↗
PubMed1991

A simple and improved method to generate human hybridomas.

By improving our previous method for production of human hybridomas, we developed a simple and remarkably efficient method for production of human hybridomas. We reformed our previous method in the following three points: (1) we added irradiated (30 Gy) myeloma cells as feeder cells to culture of fusion cells; (2) ouabain concentration in the selective medium was reduced from 2 x 10(-6) M to 1 x 10(-6) M; (3) selective medium (GIT-HAT-OL) was added to the culture after overnight cultivation of fusion cells. Consequently, we obtained higher fusion frequency (1/700 vs. 1/5500) compared with our previous method. By our present method, only so small number of human B cells (EBV-LCL) is required, so that the time necessary to establish human hybridomas is reduced.

Cell Fusion↗
PubMed1991

Prostaglandin E1 receptor from mouse mastocytoma P-815 cells couples to 60 kDa GTP-binding protein.

The stable [3H]prostaglandin E1 (PGE1)-bound receptor, which couples to 60 kDa GTP-binding protein, from membranes of mouse mastocytoma P-815 cells has been purified and characterized. When the membranes were preincubated with [3H]PGE1 for 60 min at 37 degrees C, the dissociation of the ligand from the receptor was remarkably decreased, even in the presence of GTP gamma S. The stable [3H]PGE1-bound receptor complex was solubilized with 6% digitonin. The solubilized [3H]PGE1 receptor was eluted with [35S]GTP gamma S bindings activity from an Ultrogel AcA44 column. The fractions containing activities of both [3H]PGE1 and [35S]GTP gamma S bindings were further purified by column chromatographies on wheat germ agglutinin (WGA)-agarose and phenyl-Sepharose CL-4B. The partially purified [3H]PGE1-bound receptor was affinity-labeled with [14C]5'-p-fluorosulfonylbenzoylguanosine and a protein with a molecular mass of 60 kDa was detected. These results suggest that the ligand-bound PGE1 receptor of P-815 cells associates with a novel GTP-binding protein with a molecular mass of 60 kDa.

Adenylyl Cyclases↗
PubMed1991

Immunological changes in Babesia rodhaini infected BALB/c mice after treated with anti-babesial drug; diminazene diaceturate.

A model system capable of investigating immunological changes was first established in Babesia rodhaini infected mice with an aid of a drug, diminazene diaceturate (DD). Intraperitoneal (ip) inoculation with B. rodhaini resulted in acute death in euthymic (nu/+) and athymic (nu/nu) BALB/c mice. Treatment with DD at an early stage of infection saved both mice from acute death. Parasitemia recurred in some of them but resulted in death only in nu/nu mice. A re-challenge with 10(5) parasitized erythrocytes (PE) on the surviving mice on day 28 post infection revealed resistance in nu/+ but not in nu/nu mice. The results suggested a participation of the thymus in the protective mechanisms. Immunological changes were then observed on nu/+ and nu/nu mice which were inoculated ip with 10(4)PE and treated with the drug, and then challenged with 10(5)PE ip on day 28. An antibody response was measured with immediate reaction by footpad injection of a soluble antigen of B. rodhaini and by ELISA of serum antibody using the antigen and protein A, on day 10 and later, and further a pronounced response was detected after re-challenge in nu/+ mice. No response was detected by ELISA in nu/nu mice. Delayed footpad reaction was seen in nu/+ mice by day 14 and later but it was suppressed after the re-challenge.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1990

[Intraperitoneal administration of recombinant interferon-beta for prevention in peritoneal metastasis of gastric cancer].

Recombinant interferon-beta (IFN-beta) shows a high anti-tumor effect on cancerous ascites, has little local irritative action and can safely be administered intraperitoneally. A randomized trial was conducted to evaluate the efficacy of intraperitoneal administration of IFN-beta for the purpose of preventing peritoneal metastasis of gastric cancer. The subjects selected were patients under 75 years of age who were macroscopically positive for serosal invasion and had undergone curative operation. Comparison was made by the envelope method between the cases given IFN-beta i.v. at 6 x 10(6) units continuously for 14 days from the day of operation and from 1 week after operation. MMC, 20 mg (i.v.) on the day of operation and 5-FU, 200 mg (p.o.) for 1 year from 2 weeks after operation were administered, respectively, in the two groups. The number of cases registered from March, 1987 to September, 1988 was 33, with 17 in the administration group and 16 in the control group. No difference was noted between the two groups in 3-year survival rate and disease free survival rate. As to the recurrence involving the peritoneum, however, there was a difference between the two groups, with 1 out of 6 cases in administration group and 3 out 4 cases in control group. Intraperitoneal chemotherapy can be an effective adjuvant therapy for gastric cancer, if cases are selected.

Adult↗
PubMed1990

[Effect of medroxyprogesterone acetate on the cell kinetics in primary breast cancer].

Effect of medroxyprogesterone acetate (MPA) on the breast cancer cell kinetics was investigated by flow cytometry on paraffin-embedded tissues. Nuclear DNA contents were measured in 67 primary cases. MPA, 1,2000 mg/day, was administered orally for two weeks in 12 cases (MPA group) and the remaining cases (n-MPA group) served as the controls, until the day before operation. The DNA histograms were compared between both groups. The mean percentage of G0 + G1 phase was higher and that of S-phase and G2 + M phase, lower, in the MPA group than those in the n-MPA group. Especially in estrogen receptor positive and premenopausal cases, a significant difference was present between both groups. These results suggest that MPA could inhibit DNA synthesis with a delay of the cell cycle progression in human breast cancer.

Antineoplastic Agents↗
PubMed1989

Studies on route of immunization with a mixture of killed parasites and adjuvants against Babesia rodhaini infection in mice.

A series of experiments were undertaken to determine the most effective route of immunization with a mixture of killed Babesia rodhaini antigen (S antigen) and formalin-fixed Corynebacterium parvum (Propionibacterium acnes) bacterin (CPB) against challenge infection with B. rodhaini 3 weeks later. The mice pretreated with S antigen and CPB mixture intraperitoneally, but not intramuscularly, were significantly resistant to intraperitoneal (IP) or intravenous (IV) challenge with 10(6) organisms. The survival rates were 70.0 (IP challenge) and 60.0% (IV challenge) respectively. Fairly protective activities were equally produced in mice intravenously pretreated with S antigen and CPB with survival rates of 60.0% against IV challenge, but 30% against IP. These results indicated that the IP injection of S antigen and CPB mixture is desirable route for immunization against subsequent IP or IV challenge with B. rodhaini. On the other hand, lower protective effect was reconfirmed in the mice treated with S antigen and Freund's Complete adjuvant, regardless of immunization routes in the additional experiment. The survival rates were 33.3, 14.3 and 11.8% in the intraperitoneally, intramuscularly and subcutaneously-treated mice respectively against IP challenge with 10(6) organisms.

Animals↗
PubMed1989

Artificial infection by transplantation of juvenile Fasciola worms into the abdominal cavity of rats.

Rats were successfully infected with Japanese Fasciola sp. by transplantation of juvenile worms (JW) or metacercariae (MC) into the abdominal cavity. Moreover, the rat was investigated on its suitability for different experiments with liver flukes. JWs or MCs transplanted intraperitoneally (IP) matured in the bile duct of rats. Moreover, more stable infections were established by inoculation of JWs than MCs. About 3 of 10-15 JWs transplanted into the abdominal cavity of a rat matured and laid eggs in the bile duct. The mean prepatent period was 63.5 days in the JW inoculated group. EPG values were kept constant at a level of 10(2)-10(3) about 100 to 230 days after the transplantation of JWs. The life span of Japanese liver flukes was estimated to be about 400 days in rats. From these results, it was concluded that the rat is suitable for various experiments with Fasciola sp.

Animals↗
PubMed1989

[Two cases of advanced breast cancer with distant metastasis showing long-term survival with multidisciplinary treatment including intra-arterial infusion chemotherapy].

Two cases of advanced breast cancer are reported that have had no recurrence for more than five years after multidisciplinary treatment including intra-arterial infusion chemotherapy. The first case is a 62-year-old housewife who had brain metastases. The primary lesion and distant metastases were completely responsive to irradiation combined with chemoendocrine therapy. The other one is a 38-year-old housewife with ovarian metastasis that was revealed after oophorectomy. In this case chemoendocrine therapy was added without irradiation after intra-arterial infusion chemotherapy. These cases indicate that intra-arterial infusion chemotherapy can be recommended as a component of multidisciplinary treatment even in advanced breast cancer with distant metastasis.

Adenocarcinoma, Scirrhous↗