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Biomedical subjects

T Suda

Publications and source records attributed to T Suda.

At least 775 records · Page 43Linked to original sources

[Immuno-enhancing actions of carnosine and homocarnosine].

Immuno-enhancing actions of carnosine, beta-alanine, homocarnosine, and gamma-aminobutyric acid were studied in ddY mice by evaluating plaque-forming cell reaction against sheep red blood cells. Animals were administered the test agents in prior to, or simultaneously with, various treatments that are known to reduce immune function such as administration of the anti-tumor agents, mitomycin C and 5-fluorouracil, immunosuppressant cyclophosphamide, antiinflammatory agent hydrocortisone, or cancer implantation and gamma-irradiation. Experiments were performed also in aged mice with reduced immune function. The administration of these drugs showed non-specific immuno-enhancing effects under all conditions examined and on all cell groups that may have been affected by these immunosuppressive stimulus.

Adjuvants, Immunologic↗

[Clinical evaluation of adriamycin ointment in advanced or local recurrent breast cancer].

For the purpose of local therapy for advanced and recurrent breast cancer, we have applied a new Adriamycin (ADR) ointment. This new ADR ointment has been prepared by the technique of a two-factor composite experimental design and includes PEG: 25%, CVP: 0.65%, HPC: 1.35% and ADR: 0.04%. We have applied it to three patients with advanced breast cancer and six patients with locally recurrent breast cancer. By using this ointment, the skin ulcers have become dry, bleeding has stopped and the sense of heat has been lost. In some cases, the neoplastic ulcers have diminished in size. Some problems with this ointment are easy bleeding at the time of removing the gauze and an unstable effect in diminishing the size of the neoplasm. We therefore think that the use of this ointment alone is very effective for controlling the symptoms of the local lesion, but not so effective for diminishing the size of the neoplasm.

Administration, Topical↗

In vitro study of immunoreactive corticotropin-releasing factor release from the rat hypothalamus.

Immunoreactive corticotropin-releasing factor (I-CRF) release from rat hypothalami was studied in vitro utilizing a perifusion of rat hypothalami and a rat CRF RIA. Basal release of I-CRF from the hypothalamus of adrenalectomized or hypophysectomized rats was higher than in that of normal rats. K+-induced I-CRF release was completely suppressed by omission of Ca++ from the medium. Dexamethasone suppressed I-CRF release from hypothalami, but not from median eminence (ME). C-AMP and angiotensin II had mild stimulatory effects on I-CRF release. These results suggest that 1) the feedback mechanism acts mainly on a higher level than ME, and 2) c-AMP and angiotensin II may be involved in CRF-releasing mechanism(s).

Adrenalectomy↗

The mechanism of end-organ resistance to 1 alpha,25-dihydroxycholecalciferol in the common marmoset.

The common marmoset, a New World monkey, requires a large amount of cholecalciferol (110 i.u./day per 100g body wt.) to maintain its normal growth. In a previous report, we demonstrated that the circulating levels of 1 alpha, 25-dihydroxycholecalciferol [1 alpha,25(OH)2D3] in the marmosets are much higher than those in rhesus monkeys and humans, but the marmosets are not hypercalcaemic [Shinki, Shiina, Takahashi, Tanioka, Koizumi & Suda (1983) Biochem. Biophys. Res. Commun. 14, 452-457]. To compare the effect of the daily intake of cholecalciferol, two rhesus monkeys were given a large amount of cholecalciferol (900 i.u./day per 100g body wt). Their serum levels of calcium, 25-hydroxycholecalciferol and 24R,25-dihydroxycholecalciferol were markedly elevated, but the serum 1 alpha,25(OH)2D3 levels remained within a range similar to those in the rhesus monkeys fed the normal diet (intake of cholecalciferol 5 i.u./day per 100g body wt). Intestinal cytosols prepared from both monkeys contained similar 3.5 S macromolecules to which 1 alpha,25(OH)2D3 was bound specifically. However, the cytosols from the marmosets contained only one-sixth as many 1 alpha,25(OH)2D3 receptors as those from the rhesus monkeys. Furthermore, the activity of the 1 alpha,25(OH)2D3-receptor complex in binding to DNA-cellulose was very low in the marmosets. These results suggest that the marmoset possesses an end-organ resistance to 1 alpha,25(OH)2D3 and is a useful animal model for studying the mechanism of vitamin D-dependent rickets, type II.

Animals↗

Induction of spermidine N1-acetyltransferase by 1 alpha,25-dihydroxyvitamin D3 as an early common event in the target tissues of vitamin D.

We have reported that the duodenal ornithine decarboxylase activity and the tissue content of putrescine increase markedly after a single intravenous injection of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) into vitamin D-deficient chicks (Shinki, T., Takahashi, N., Miyaura, C., Samejima, K., Nishii, Y., and Suda, T. (1981) Biochem. J. 195, 685-690). In the present study, we examined in the same experimental system the effect of 1 alpha,25(OH)2D3 on the activity of duodenal spermidine N1-acetyltransferase, a rate-limiting enzyme catalyzing the conversion from spermidine to putrescine. The duodenal spermidine N1-acetyltransferase activity began to increase 30 min after a single intravenous injection of 625 ng of 1 alpha,25(OH)2D3 and attained a maximum in 2 h. As little as 1.25 ng of 1 alpha,25(OH)2D3 induced a small but significant increase in the spermidine N1-acetyltransferase activity, and the maximal response was obtained by 125 ng of the vitamin. The dose levels of 1 alpha,25(OH)2D3 required to induce duodenal spermidine N1-acetyltransferase activity were only one-tenth as much as those required to induce ornithine decarboxylase activity. The spermidine N1-acetyltransferase activity was induced commonly in the target tissues of vitamin D, whereas ornithine decarboxylase activity occurred only in intestine. The 1 alpha,25(OH)2D3-induced spermidine N1-acetyltransferase activity was greatly inhibited by prior administration of actinomycin D or cycloheximide. When 30,000 X g intestinal supernatants were incubated with [14C]spermidine, [14C]putrescine was formed. These results clearly indicate that the induction of spermidine N1-acetyltransferase activity is the 1 alpha,25(OH)2D3-induced earliest de novo synthesis in several proteins induced by the vitamin reported to date and that the 1 alpha,25(OH)2D3-induced duodenal synthesis of putrescine occurs by the pathways from both ornithine and spermidine.

Acetyltransferases↗

Proliferation and differentiation in culture of mast cell progenitors derived from mast cell-deficient mice of genotype W/Wv.

Mice of genotype W/Wv have less than 1% of normal mast cells in the skin, stomach, and cecum. In order to further clarify the mechanism of this deficiency, we studied committed mast cell progenitors and multipotent progenitors, which are capable of mast cell differentiation in clonal culture. The relative concentration of mast cell progenitors in the bone marrow, spleen, and peripheral blood of W/Wv mice was similar to that of +/+ mice. However, the cellularity of the marrows of W/Wv mice was 54% of that of their normal littermates. Identification of mast cells was established by metachromatic staining with toluidine blue, transmission electron microscopy, and demonstration of membrane receptors for immunoglobulin E. The time course of colony formation and the morphology of W/Wv mast cell colonies in culture was identical to that of normal littermates. The percentages of mast cells in individual multi-lineage colonies were extremely variable. The histamine content of mast cells derived from W/Wv mice was similar to that of mast cells from +/+ mice. These studies demonstrated the normal capacity for differentiation and proliferation in culture of mast cell progenitors from W/Wv mice.

Animals↗

Permissive role of interleukin 3 (IL-3) in proliferation and differentiation of multipotential hemopoietic progenitors in culture.

We studied the effects of interleukin-3 (IL-3) on colony formation by hemopoietic progenitors in methylcellulose cultures of spleen cells from 5-fluorouracil (FU)-treated mice. Purified IL-3 supported the growth of various types of multilineage colonies including blast cell colonies. The types of colonies were similar to those supported by pokeweed-mitogen spleen cell conditioned medium (PWM-SCM), except that IL-3 supported eosinophil and neutrophil expression better. Delayed addition of IL-3 to cultures 7 days after cell plating decreased the number of colonies to one-half the number in cultures with IL-3 added on day 0. It did not alter the proliferative and differentiation characteristics of late emerging multipotential blast cell colonies. These observations suggest that IL-3 does not trigger hemopoietic progenitors into active cell proliferation but is necessary for their continued proliferation. This permissive role of IL-3 is consistent with a stochastic model of stem cell proliferation which features random entry into cell cycle. IL-3 also supported the growth of multilineage colonies from single cells isolated from blast cell colonies by micromanipulation. This result shows that IL-3 acts directly on multipotential progenitors. Analysis of colonies derived from paired progenitors revealed disparate lineage expression and was in accordance with the stochastic model of stem cell differentiation.

Animals↗

1 alpha,25-dihydroxyvitamin D3 receptors and their action in embryonic chick chondrocytes.

The role of vitamin D in the maturation of epiphyseal chondrocytes was investigated in the developing chick embryo. Cartilage tissues were divided into two parts: resting cartilage and growth cartilage. A cytosol component to which 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) is specifically bound first appeared in the growth cartilage on day 15, rapidly increased, and attained a maximum on day 19. The calcium content of the growth cartilage also began to increase on day 15 and continued to increase in parallel with the 1 alpha,25(OH)2D3 receptor levels. Glycosaminoglycan (GAG) synthesis by the growth cartilage cells increased from day 11-17 and rapidly declined thereafter reciprocally with the increase in calcium and receptor levels. In the resting cartilage, no cytosol receptor for 1 alpha,25(OH)2D3 was detected up to hatching time. The calcium content and GAG synthesis in the resting cartilage were very low and did not change appreciably throughout development. No receptor-like macromolecule for 24R,25-dihydroxyvitamin D3 (24R,25(OH)2D3) was recognized in either the resting or growth cartilage. 1 alpha,25(OH)2D3 added to the culture of chondrocytes from the epiphyseal growth cartilage inhibited GAG synthesis and stimulated its release from the cell layer into the medium in a dose-dependent manner. These in vitro effects of 1 alpha,25(OH)2D3 were not observed in chondrocytes obtained from 13-day-old growth cartilage and 19-day-old resting cartilage. 25-Hydroxyvitamin D3 and 24R,25(OH)2D3 had no effect on chondrocytes in any of the preparations. These results suggest that 1 alpha, 25 (OH)2D3 is directly involved in the maturation of chondrocytes and possibly in the calcification of growth cartilage.

Animals↗

Capillary permeability-increasing enzyme from the venom of Agkistrodon caliginosus (kankoku-mamushi): activity due to the release of peptide material from a protein in bovine plasma.

When a mixture of the purified capillary permeability-increasing enzyme from A. caliginosus venom and bovine plasma or heated bovine plasma was injected into the depilated skin of the back of a rabbit, the capillary permeability-increasing activity was much greater than that induced by injection of the enzyme alone. The substance which increases capillary permeability was extracted from the incubated mixture of bovine plasma and enzyme with 50-70% ethanol. Its activity was lost when treated with carboxypeptidase A. Thus, it is supposed that the increase in capillary permeability induced by the enzyme is due to a low molecular weight peptide released from a protein in bovine plasma by the action of the enzyme. No liberation by the enzyme of histamine or anaphylatoxins of the complement system was found.

Animals↗

Effects of aging on the lipid order and composition of rat adipocyte ghosts.

An analysis of the cholesterol/phospholipid ratio of adipocyte ghosts from rat epididymal fat pads shows a significant increase with age (P less than 0.005). An attempt to correlate these changes with the order of the lipid matrix was made using the stearic acid spin label 2-(3-carboxypropyl)-4, 4-dimethyl-2-tridecyl-3-oxazolidinyloxyl [I(12,3)]. Although order was negatively correlated with temperature in preparations from both 6- and 24-month-old rats, no effect of age could be detected.

Adipose Tissue↗

Differences in the side-chain metabolism of vitamin D3 between chickens and rats.

In vitro metabolism of 25-hydroxy-24-oxovitamin D3 was studied in kidney homogenates from vitamin D-supplemented chickens and rats. In chicken homogenates, 25-hydroxy-24-oxovitamin D3 was converted predominantly to 23,25-dihydroxy-24-oxovitamin D3, 24,25-dihydroxyvitamin D3, and 23,24,25-trihydroxyvitamin D3. In rat homogenates, 25-hydroxy-24-oxovitamin D3 was not converted to either 24,25-dihydroxyvitamin D3 or 23,24,25-trihydroxyvitamin D3, but it was converted to 23,25-dihydroxy-24-oxovitamin D3 and 23-hydroxy-24,25,26,27-tetranorvitamin D3. The latter metabolite was not produced by the chicken preparations. The stereochemical configuration at C-24 of the 24,25-dihydroxyvitamin D3 produced by chicken homogenates was determined to be S. This contrasts with the R configuration of 24,25-dihydroxyvitamin D3 produced by 24-hydroxylation of 25-hydroxyvitamin D3. These results suggest that chickens have an enzyme that can reduce the 24-oxo group to 24S-hydroxyl group, whereas rats do not.

Animals↗

Immunoreactive corticotropin-releasing factor in human plasma.

Plasma immunoreactive corticotropin-releasing factor (I-CRF) levels were determined by using a human CRF radioimmunoassay and an immunoaffinity procedure. The basal plasma I-CRF level in normal subjects was 6 +/- 0.5 pg/ml (mean +/- SD). We found that most plasma I-CRF levels were affected by stress, negative feedback, and circadian rhythm. Basal I-CRF levels were high in patients with Addison's disease, Nelson's syndrome, hypopituitarism stemming from pituitary macroadenoma, and CRF- and adrenocorticotropic hormone-producing tumors. A very low, but significant, amount of I-CRF was detected (1-3 pg/ml) in patients with Cushing's syndrome, in corticosteroid-treated patients, and in a patient with hypothalamic hypopituitarism. These results suggest that a major component of plasma I-CRF is of hypothalamic origin, however, other extrahypothalamic tissues cannot be ruled out as a minor source of plasma I-CRF.

Adrenal Gland Diseases↗

The use of the corticotropin-releasing hormone test to monitor the recovery of patients with Cushing's disease or Cushing's syndrome due to an adrenal adenoma after adenomectomy.

Six patients with Cushing's disease and three with Cushing's syndrome due to an adrenal adenoma were monitored after their adenomectomy with the corticotropin-releasing hormone test to evaluate the progress of recovery of their pituitary adrenal function. Before surgery the patients with Cushing's disease showed either high, normal or low responses of plasma ACTH and cortisol to 100 micrograms synthetic ovine corticotropin-releasing hormone (CRH) administered intravenously, whereas all three patients with Cushing's syndrome due to an adrenal adenoma showed no response of plasma ACTH or cortisol to CRH. One or two months after surgery, the patients who had Cushing's disease had low levels of basal plasma ACTH and cortisol and their responses to CRH were extremely low. However, the same patients were tested later, it was found that their responses to CRH gradually increased and reached normal ranges approximately within one year after tumor removal, which coincided with the overall improvement in their clinical signs and symptoms due to adrenal insufficiency. In contrast, the recovery of the pituitary adrenal function in patients who had Cushing's syndrome due to an adrenal adenoma was not complete even one year after surgery. Thus the corticotropin-releasing factor test is a useful criteria to evaluate the recovery of the pituitary adrenal function in these patients after surgery, since the responses of plasma ACTH and cortisol to the administered CRH are parallel with the improvements in clinical signs and symptoms due to adrenal insufficiency in patients with Cushing's disease.

Adenoma↗

Clonal analysis of basophil differentiation in bone marrow cultures from a Down's syndrome patient with megakaryoblastic leukemia.

We present the in vitro differentiation of marrow cells from a patient with Down's syndrome accompanied by megakaryoblastic leukemia into basophils in the presence of phytohemagglutinin-stimulated leukocyte conditioned medium, using a liquid culture and methylcellulose culture system. Identification of basophils was established by metachromatic staining with toluidine blue, transmission electron microscopy, and the presence of histamine. However, these basophils did not release histamine in response to calcium ionophore or chemotactic peptide. Samples from suspension cultures that contained 90% basophils showed chromosomal markers characteristic of leukemic cells (48, XY, +11, +21, t(1;15)) in all examined mitoses. The cellular composition of leukemic colonies grown in methylcellulose culture from single cells was studied using the micromanipulation technique. High plating efficiency and extreme predominance of basophil colonies were observed. In a total 137 cultures, 79 revealed colony growth. Of 59 colonies that were analyzed by cytologic examination, 46 were pure basophil colonies. These basophil colonies showed disperse morphology, similar to that of a normal basophil colony. The clonality of the basophil colonies and skewing of lineage expression were documented from leukemic single-cell cultures. These data showed that leukemic cells have the capacity for differentiation into some lineages that are not expressed in vivo.

Basophils↗

Regulation of melanin synthesis of B16 mouse melanoma cells by 1 alpha, 25-dihydroxyvitamin D3 and retinoic acid.

Melanin synthesis of B16 mouse melanoma cells was found to be stimulated dose and time dependently by 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], the hormonal form of vitamin D3. The stimulation of melanogenesis resulted from an increase in the activity of tyrosinase, a key enzyme in melanin synthesis. The minimum dose required for this stimulation was as low as 0.05 ng/ml, or 0.12 nM, a physiological level of plasma 1 alpha,25(OH)2D3. The stimulation by 1 alpha,25(OH)2D3 was specific; other derivatives of vitamin D3 caused no stimulation at a concentration of 500 ng/ml. When the cells were plated on agar plates, the proportion of dark or black colonies was not increased by the exposure to 1 alpha,25(OH)2D3. Furthermore, this compound did not induce melanin synthesis of an amelanotic variant. Thus, its stimulatory effect seemed to be due to stimulation of melanin synthesis of melanotic cells, rather than to conversion of amelanotic clones to melanotic ones. 1 alpha,25(OH)2D3 did not induce intracellular cyclic adenosine 3':5'-monophosphate, while cholera toxin induced cyclic adenosine 3':5'-monophosphate and stimulated melanin synthesis and tyrosinase activity much more than did 1 alpha,25(OH)2D3, suggesting that 1 alpha,25(OH)2D3 stimulates melanin synthesis by a cyclic adenosine 3':5'-monophosphate-independent mechanism. B16 melanoma cells contained specific receptors for 1 alpha,25(OH)2D3. Scatchard plot analysis revealed two types of receptor; the high-affinity receptor had a Kd of 18.3 pM and an Nmax of 10.6 fmol/mg of protein. The specificity of receptor binding was demonstrated by studies showing that, for 50% displacement of 1 alpha,alpha,25(OH)2D3 binding, other derivatives were required at 500 times higher concentrations or more. In contrast to 1 alpha,25(OH)2D3, retinoic acid inhibited melanin synthesis and tyrosinase activity of B16 melanoma cells dose and time dependently. On simultaneous treatment, 1 alpha,25(OH)2D3 and retinoic acid caused mutual interference, and a balance between their respective stimulating and inhibitory effects was obtained at a molar ratio of 10:1; i.e., with 10 nM 1 alpha,25(OH)2D3 and 1 nM retinoic acid.

Animals↗

Alternative differentiation of human promyelocytic leukemia cells (HL-60) induced selectively by retinoic acid and 1 alpha,25-dihydroxyvitamin D3.

Induction of hematopoietic differentiation was investigated in human promyelocytic leukemia cells [HL-60] using two lipophilic vitamins, retinoic acid and 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3]. Both vitamins suppressed proliferation and induced differentiation of HL-60 cells, but 1 alpha,25(OH)2D3 was 70- to 100-fold more potent than was retinoic acid on a molar basis. Simultaneous treatment with suboptimal concentrations of 1 alpha,25(OH)2D3 (0.12 to 1.2 nM) and retinoic acid (10 to 100 nM) showed additive effects in reducing nitroblue tetrazolium, a common marker for monocyte-macrophage and granulocyte differentiation. For the study of alternative differentiation of the cells by the two vitamins, we used monoclonal antibodies specific for either human monocyte-macrophages or granulocytes and other markers specific for macrophage differentiation such as alpha-naphthyl acetate esterase activity and adherence to the dish surface. HL-60 cells were induced to differentiate alternatively into macrophages by 1 alpha,25(OH)2D3 or into granulocytes by retinoic acid. When HL-60 cells were treated with various concentrations of 1 alpha,25(OH)2D3 (1.2 to 120 nM) in the presence of 1000 nM retinoic acid which is a concentration sufficient to induce maximal granulocyte differentiation, the appearance of the markers for monocyte-macrophage differentiation by 1 alpha,25(OH)2D3 was not at all affected by the retinoic acid. These results indicate that 1 alpha,25(OH)2D3 and retinoic acid have additive effects in inducing differentiation of HL-60 cells, but monocyte-macrophage differentiation by 1 alpha,25(OH)2D3 occurs much more readily than does granulocyte differentiation by retinoic acid.

Calcitriol↗