Search PubMed⌕ Search

Biomedical subjects

Y Kitamura

Publications and source records attributed to Y Kitamura.

At least 523 records · Page 29Linked to original sources

Survey of Hanganutziu and Deicher antibodies in operated patients.

The appearance of Hanganutziu and Deicher (HD) antibody in the sera of patients suffering from various diseases, including malignancies of some organs and liver disorders, was investigated by enzyme-linked immunosorbent assay using N-glycolylneuraminyl-lactosylceramide (HD3) and 4-O-acetyl-HD3 as the antigenic molecules. More than 25% of sera from patients suffering from malignancies, cholelithiasis and liver cirrhosis had HD antibody, whereas none of 41 sera from healthy persons had HD antibody. The percentage of HD antibody-positive patients was similar in stages I, II and III of gastric cancer and recurrence cases. Antibody titers of the positive patients in each stage were also not different from those in each other stage. These results indicated that HD antigenic expression on cancerous tissue is not dependent on the cancerous malignancy. The HD antibody level was elevated after surgical removal of cancerous tissues in 5 of 6 patients examined, indicating that tumor growth absorbed the serum antibody. Serum antibody against 4-O-acetyl-HD3 was detected independently of HD3 antibody in some cases; however, in most cases, correlation between the two antibody titers was observed.

Aged↗

Biologically active kit ligand growth factor is produced by mouse Sertoli cells and is defective in SId mutant mice.

In order to define the role of Kit ligand (KL) growth factor encoded at the mouse steel (SI) locus in spermatogenesis, we have examined its production in Sertoli cells. As a measure KL growth factor bioactivity, the ability to support proliferation and maintenance of mast cells was used in co-culture with primary mouse Sertoli cells. On the sertoli cells derived from +/+ and Wv/Wv mice, +/+ mast cells proliferated and were supported for more than 2 weeks, but not W/Wv mast cells. In contrast, Sertoli cells from SId/SId mice could not support +/+ mast cell proliferation under similar conditions. The supportive effect required close-range interaction of Sertoli cells with cultured mast cells. These results indicate that Sertoli cells derived from +/+ and Wv/Wv but not SId/SId mutant mice produce biologically active KL growth factor as a membrane-bound form. The biologically active KL of Sertoli cells may also play an important role in germ cell growth and differentiation.

Animals↗

Effects of nebracetam (WEB 1881 FU), a novel nootropic, as a M1-muscarinic agonist.

We here investigated the effects of nebracetam (WEB 1881 FU 4-aminomethyl-1-benzylpyrrolidin-2-one-hemifumarate), a novel pyrrolidinone nootropic, on the rise of intracellular Ca2+ concentration ([Ca2+]i) in Jurkat cells, a human leukemic T cell line. Nebracetam induced a rise of [Ca2+]i in the medium with 1 mM Ca2+ and without Ca2+ (plus 1 mM EGTA). The nebracetam-induced [Ca2+]i rise was blocked by atropine greater than pirenzepine greater than AF-DX 116. From these results, nebracetam seems to act as an agonist for human M1-muscarinic receptors.

Animals↗

Effects of bifemelane hydrochloride, a brain function improver, on muscarinic receptors in the CNS of senescence-accelerated mouse.

Senescence-accelerated mouse (SAM-P/8) is known as a murine model of aging and memory dysfunction, compared with control mouse (SAM-R/1). In the hippocampus of 9-month-old SAM-P/8, the Bmax of [3H]QNB binding was decreased compared with that of SAM-R/1 at the same age. Single and repeated administrations of bifemelane to SAM-P/8 induced an increase in the Bmax of [3H]QNB binding in the hippocampus. From these results, bifemelane seems to exert pharmacological effects through possible activation of the cholinergic system in the hippocampus of SAM.

Aging↗

Bone marrow-derived cultured mast cells and peritoneal mast cells as targets of a growth activity secreted by BALB/3T3 fibroblasts.

When fibroblast cell lines were cultured in contact with bone marrow-derived cultured mast cells (CMC), both NIH/3T3 and BALB/3T3 cell lines supported the proliferation of CMC. In contrast, when contact between fibroblasts and CMC was prohibited by Biopore membranes or soft agar, only BALB/3T3 fibroblasts supported CMC proliferation, suggesting that BALB/3T3 but not NIH/3T3 cells secreted a significant amount of a mast cell growth activity. Moreover, the BALB/3T3-derived growth activity induced the incorporation of [3H]thymidine by CMC and the clonal growth of peritoneal mast cells in methylcellulose. The mast cell growth activity appeared to be different from interleukin 3 (IL-3) and interleukin 4 (IL-4), because mRNAs for these interleukins were not detectable in BALB/3T3 fibroblasts. Although mast cells are genetically deficient in tissues of W/Wv mice, CMC did develop when bone marrow cells of W/Wv mice were cultured with pokeweed mitogen-stimulated spleen cell-conditioned medium. Because BALB/3T3 fibroblast-conditioned medium (BALB-FCM) did not induce the incorporation of [3H]thymidine by W/Wv CMC, the growth activity in BALB-FCM appeared to be a ligand for the receptor encoded by the W (c-kit) locus. Because CMC and peritoneal mast cells are obtained as homogeneous suspensions rather easily, these cells may be potentially useful as targets for the fibroblast-derived mast cell growth activity.

Animals↗

Effects of culture matrix on differentiation of murine mast cells.

The effects of culture matrix (agar, collagen, and methylcellulose) on differentiation of mast cells were investigated. Because berberine sulfate-positive colonies in agar were composed of macrophages but not of mast cells, the naphthol AS-D chloroacetate esterase reaction was used to identify mast-cell colonies. When bone marrow cells of WBB6F1 mice were cultured with conditioned media containing interleukin 3 (IL-3), numbers of mast-cell colonies were greater in collagen cultures than in agar and methylcellulose cultures. Electron microscopic examination revealed that mast cells developing in collagen and agar cultures were more mature than those developing in methylcellulose cultures. However, when bone marrow-derived cultured mast cells or purified peritoneal mast cells were plated, the efficiency of colony formation and size of colonies were comparable among agar, collagen, and methylcellulose cultures. Therefore, all three matrices tested had similar effects on the proliferation of mast cells. Collagen appeared to be suitable for differentiation of bone-marrow precursors and their maturation. Agar appeared to be suitable only for maturation.

Agar↗

Proliferative potential of murine peritoneal mast cells after degranulation induced by compound 48/80, substance P, tetradecanoylphorbol acetate, or calcium ionophore A23187.

Proliferative potential of degranulated mast cells was investigated. Mast cells were collected from the peritoneal cavity of mice, and degranulation was induced by compound 48/80, substance P, 12-O-tetradecanoylphorbol 13-acetate (TPA), or calcium ionophore A23187. The potentiality of colony formation in methylcellulose was not reduced by treatment of various concentrations of compound 48/80, substance P and TPA. When degranulation was induced by compound 48/80, substance P or TPA, proportion of highly degranulated mast cells containing less than five granules was rather small. In contrast, considerable proportion of highly degranulated mast cells was obtained after the treatment with the low concentration (0.1 microgram/ml) of A23187. These highly degranulated mast cells, which were individually picked up by the micromanipulator, proliferated not only in methylcellulose but also in the skin of mast cell-deficient WBB6F1-W/Wv mice. Inasmuch as we have already shown the proliferation of IgE-sensitized and Ag-stimulated mast cells, degranulated mast cells appear to retain the proliferative potential in general.

Animals↗

Production, purification and characterization of biologically active recombinant human nerve growth factor.

The human NGF gene was isolated and inserted downstream from murine leukemia virus LTR in a plasmid having dihydrofolate reductase cDNA. The expression plasmid was introduced into CHO cells. Selection of the transformants for the resistance to methotrexate gave a CHO cell line which produced human NGF at a level of 4 mg/L in the culture medium. The recombinant human NGF was purified to near homogeneity from the culture supernatant. The NH2-terminal amino acid sequence, the COOH-terminal amino acid (Ala), and the amino acid composition of the human NGF were identical to those deduced from the nucleotide sequence of the human NGF gene. The recombinant human NGF was composed of 120 amino acid residues. Three disulfide linkages were determined to be Cys15-Cys80, Cys-58-Cys108, and Cys68-Cys110; the locations were identical to those in the mouse 2.5S NGF molecule. The specific biological activity of the recombinant human NGF was comparable with that of authentic mouse 2.5S NGF as determined by stimulation of neurite outgrowth from PC12 cells.

Amino Acid Sequence↗

Possible involvement of different GTP-binding proteins in noradrenaline- and thrombin-stimulated release of arachidonic acid in rabbit platelets.

Noradrenaline (NA) stimulated the release of arachidonic acid (AA) from the [3H]AA-labelled rabbit platelets via alpha 2-adrenergic receptors, since the effect of NA was inhibited by yohimbine. The stimulatory effect of NA in digitonin-permeabilized platelets was completely dependent on the simultaneous presence of GTP and Ca2+. The NA- and thrombin-stimulated releases of AA were markedly decreased by the prior ADP-ribosylation of the permeabilized platelets with pertussis toxin. Antiserum directed against the pig brain Go (a GTP-binding protein of unknown function), recognizing both alpha 39 and beta 35,36 subunits, but not alpha 41, of pig brain, reacted with 41 kDa and 40 kDa bands, with not one of 39 kDa, in rabbit platelet membranes. Anti-Go antiserum inhibited guanosine 5'-[gamma-thio]triphosphate-, A1F4(-)-, NA- and thrombin-stimulated AA releases in the membranes. Although the effect of thrombin was inhibited by low concentrations of anti-Go antiserum, high concentrations of the antiserum was needed for inhibition of the NA effect. Antiserum directed against the pig brain G1 (inhibitory G-protein), recognizing both alpha 41 and beta 35,36 subunits, but not alpha 39, of pig brain, reacted with the 41 kDa band in platelets. Anti-G1 antiserum inhibited only the effect of NA. Reconstitution of the platelet membranes ADP-ribosylated by pertussis toxin with Go, not Gi, purified from pig brain restored the thrombin-stimulated release of AA. In contrast, reconstitution of those membranes with Gi, not Go, restored the NA-stimulated release of AA. These results indicate that different GTP-binding proteins, Gi- and Go-like proteins, may be involved in the mechanism of signal transduction from alpha 2-adrenergic receptors and thrombin receptors to phospholipase A2 in rabbit platelets.

Aluminum↗

Inhibitory effect of androgen on cell death of mouse uterine epithelium.

The protective effect of androgen against the cell death of mouse uterine epithelium was evaluated by examining the retention of 5'-[125I]iodo-2'-deoxyuridine ([125I]IdUrd) incorporated into the whole uterus and the apoptotic index (percentage of the apoptotic cells to the total cells) which is a good index of physiological cell death. Castrated adult female mice were daily injected with oestradiol-17 beta for 3 days, followed by the injection of [125I]IdUrd. Thereafter, these mice were daily injected with only the vehicle or 5 alpha-dihydrotestosterone (DHT), and the 125I-radioactivity retained in the whole uterus was determined. When only the vehicle was injected, the 125I-radioactivity retained in the whole uterus rapidly decreased but injections of DHT reduced the loss of 125I-radioactivity. The effect of DHT on the retention of 125I-radioactivity depended on doses of DHT and was abolished by the pure antiandrogen, flutamide. The apoptotic index of uterine cells was examined by a similar experimental protocol, but without an injection of [125I]IdUrd. Injections of only the vehicle caused marked increases in the apoptotic indices of both luminal and glandular epithelia, but injections of DHT decreased them significantly. The apoptotic index of stroma was not affected by the injection of DHT. The present results indicated that androgen reduces the cell death of mouse uterine epithelium through the androgen receptor.

Animals↗

Mechanism of mast cell deficiency in mutant mice of mi/mi genotype: an analysis by co-culture of mast cells and fibroblasts.

Mutant mice of mi/mi genotype are osteopetrotic and are deficient in mast cells. The osteopetrosis of mi/mi mice can be cured by bone marrow transplantation from congenic normal (+/+) mice, and therefore, the cause of the osteopetrosis is attributed to a defect of osteoclasts. Since both osteoclasts and mast cells are the progeny of multipotential hematopoietic stem cells, we examined whether mast cells were defective in mi/mi mice. In spite of the deficiency of mast cells in tissues of mi/mi mice, mast cells did develop when spleen cells of mi/mi mice were cultured with pokeweed mitogen-stimulated spleen cell conditioned medium (PWM-SCM). The proliferative response of cultured mast cells (CMC) derived from mi/mi mice to PWM-SCM was comparable with that of CMC from +/+ mice. In contrast, when CMC were co-cultured with the NIH/3T3 fibroblast cell line in culture medium lacking PWM-SCM, only +/+ CMC entered into the S phase of the cell cycle and were maintained; mi/mi CMC gradually disappeared. Moreover, fibroblasts derived from the skin of mi/mi mice normally supported the proliferation of +/+ CMC. Thus, the mast cell deficiency of mi/mi mice appears to be due to the inability of mi/mi mast cells to respond to the proliferative stimulus presented by fibroblasts.

Animals↗

Necessity of IgE antibodies and mast cells for manifestation of resistance against larval Haemaphysalis longicornis ticks in mice.

Genetically mast cell-deficient WBB6F1-W/Wv mice showed an apparent defect in manifestation of the resistance against larval Haemaphysalis longicornis ticks, but their serum IgE levels increased more than 100-fold after the second tick infestation. Immune sera obtained from the WBB6F1-W/Wv mice were adoptively transferred to the other WBB6F1-W/Wv mice which had received intracutaneous injections of WBB6F1-+/+ mouse-derived cultured mast cells. Because the resistance against ticks was detectable only when both mast cells and IgE antibodies were available, immediate hypersensitivity reaction appeared to have a physiologic role in the manifestation of the resistance against H. longicornis ticks.

Animals↗

Homologous recombination in a mammalian plasmid.

Bovine papillomavirus (BPV) shuttle vectors replicate as a circular plasmid in mouse cell nuclei without impairing host cell viability. We used these vectors to analyze homologous recombination in mammalian cells. When several BPV-based plasmids carrying direct repeats were introduced into C127 cells, we detected many recombinant plasmid molecules that have lost the sequence between the repeats. Many recombinant type molecules as well as parental type molecules were detected in all the cell clones isolated for analysis. Sequencing after rescue of the plasmid in Escherichia coli showed that most of the recombinants were from accurate homologous recombination. When the repeats on the plasmid were in inverted orientation, no crossing-over type products were detected. We discuss possible mechanisms that explain these features.

Animals↗

Administration of 8-methoxypsoralen and ultraviolet A irradiation (PUVA) induces turnover of mast cells in the skin of C57BL/6 mice.

Administration of 8-methoxypsoralen followed by ultraviolet A irradiation (PUVA treatment) has been used as a therapy for urticaria pigmentosa. The effect of PUVA treatment on cutaneous mast cells in mice was investigated by using giant granules of mast cells from C57BL/6-bgJ/bgJ (Chediak-Higashi syndrome) mice as a marker. C57BL/6-(+)/+ mice were lethally irradiated and rescued by bone marrow transplantation from C57BL/6-bgJ/bgJ mice. In the radiation chimeras, mast cells in the skin were of +/+ type and mast-cell precursors migrating in the bloodstream were bgJ/bgJ. When PUVA treatment was applied to the skin of the radiation chimeras, the total number of mast cells continued to decrease until the third week after the treatment and then recovered to pre-treatment levels. The initial reduction was attributed to the decrease of +/(+)-type mast cells, and the subsequent recovery to be as a result of the increase of bgJ/bgJ-type mast cells. This observation may explain the fact that the therapeutic effect of PUVA treatment is transient. Symptoms of urticaria pigmentosa become manifest after the cessation of PUVA treatment probably because new mast cells differentiate from bone marrow-derived precursors.

Animals↗

Squamous cell carcinoma of the esophagus with cartilaginous metaplasia at metastatic lesions.

Subdermal metastatic nodules in a 62-year old male patient with esophageal carcinoma contained both carcinomatous and chondroid areas. The carcinomatous areas showed the histology of poorly differentiated squamous cell carcinoma, and light microscopically an apparent transition could be traced from carcinomatous cells to chondroid cells. In the chondroid cells, the characteristics of chondrocytes were demonstrated by light microscopic, electron microscopic, histochemical and immunohistochemical studies, although nuclear atypism was evident, suggesting their malignancy. Furthermore, immunohistochemical studies showed that some chondroid cells contained both keratin proteins and squamous cell carcinoma antigen, which were also found in the carcinomatous cells. These findings together with the light microscopic observations suggest that chondroid cells are derived from squamous cell carcinoma cells.

Carcinoma, Squamous Cell↗