[Myelodysplastic syndromes as preleukemic states].
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Biomedical subjects
Publications and source records attributed to S Kitagawa.
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The effects of long-chain unsaturated fatty acids such as linoleic acid on bovine platelets were examined. Not only linoleic acid, but also oleic and linolenic acid, at just below the concentrations causing marked cell lysis, induced an absorbance decrease of the platelet suspension in the presence of Ca2+. Since this absorbance decrease was reversed by the addition of EDTA and moreover aggregate formation was found by macroscopic and microscopic observation, it was concluded that unsaturated fatty acids at just below their lytic concentrations caused platelet aggregation. Unsaturated fatty acids also caused release of adenine nucleotides, but there was a lag time between the release and the aggregation, just as with ADP-induced release, suggesting that the aggregation was independent of the release of ADP. It was revealed that this activation of platelets by unsaturated fatty acids was caused by marked Ca2+ uptake into the cytoplasm, resulting from significant membrane perturbation.
The inhibitory effects of saturated fatty acids with 4 to 18 carbon atoms on ADP-induced aggregation of bovine platelets were investigated. The inhibitory effects of the acids increased with increase of their alkyl chain length up to C14. On the other hand, from C16 the inhibitory effects tended to decrease with increase of chain length, and stearic acid (C18) was not inhibitory. There was a linear relationship between the inhibitory effects and alkyl chain lengths up to C12. This linear relation and the slope of the linear regression line suggested that the inhibitory effects of the acids depended on their partition into the membrane. The fatty acids decreased the fluorescence of the surface charge probe 2-p-toluidinylnaphthalene-6-sulfonate, indicating that they increased the negative charge on the membrane surface. The relative effects of the acids on the fluorescence were consistent with their relative inhibitory effects on aggregation. These results suggest that the inhibition of platelet aggregation by saturated fatty acids is due to a change in the membrane surface charge of the platelet plasma membrane.
The effects of four types of reagents--a stimulant analog (ATP), reagents increasing cAMP (theophylline and (-)-isoproterenol), Ca2+ blockers (chlorpromazine, procaine, dibucaine and tetracaine) and nonspecific membrane-reactive reagents (n-butanol, n-hexanol and linoleate) - on ADP-induced Ca2+ mobilization and aggregation of platelets were investigated. All the reagents tested inhibited the aggregation. Of these reagents, those increasing cAMP and the stimulant analog inhibited the aggregation at least partly by inhibiting Ca2+ mobilization, whereas Ca2+ blockers and nonspecific membrane-reactive reagents must have inhibited the aggregation by different mechanisms, because they had: (1) no effect on ADP-induced Ca2+ mobilization, (2) accelerated it, or (3) themselves stimulated Ca2+ mobilization. The results showed that the inhibitory effects of Ca2+ blockers were at least partly due to competition with Ca2+ for binding sites on the outside of the membrane, whereas the effects of the nonspecific membrane-reactive reagents tested were due to membrane perturbation.
The effects of four alcohols--n-propyl, n-butyl, n-amyl and n-hexyl alcohol--on the ADP-induced aggregation of gel-filtered bovine platelets were examined. All four alcohols inhibited the aggregation, the order of their effects being n-propyl less than n-butyl less than n-amyl less than n-hexyl. Comparison of the inhibitory effects of the alcohols with their physico-chemical properties showed that their degrees of inhibition depended on their hydrophobicities. Moreover, it was suggested that their interaction with the lipid layer of the membrane was important for the inhibition. Studies on the effects of alcohols on the fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene-labeled platelets showed that the membrane fluidity of the platelets increased in the same concentration range in which aggregation inhibition was observed. Since the alcohols inhibited aggregation without affecting Ca2+ mobilization in the platelets, as revealed in this study, it was concluded that inhibition of platelet aggregation was due to perturbation of membrane lipids by the alcohols. This hypothesis is supported by several recent studies on the effects of cholesterol and cations, which suggest that a relatively rigid membrane favors platelet aggregation.
Regenerating areas of human lungs in pulmonary fibrosis were observed electron microscopically, and peroxidatic activity of catalase in lung peroxisomes were demonstrated cytochemically. Proliferation of Type II cells was prominent there, and some of the cells extended their cytoplasms to cover the denuded basement membrane. Unusual intermediate cells between Type II and Type I cells were observed. The extension of cytoplasmic processes with new generation of pinocytotic vesicles strongly suggested a Type I cell profile. However, catalase-positive peroxisomes were found in these cells simultaneously. From these results it was concluded that Type I cells may originate from Type II cells in human lungs as they do in experimental animals.
The natural killer (NK)-interferon (IFN) system was investigated in patients with preleukaemic states. Endogenous NK cell activity was markedly reduced in all 12 patients studied. No significant correlation was observed between the activity of NK cells and the percentage of blast cells in the peripheral blood or bone marrow. The frequency of large granular lymphocytes (LGL) in lymphocytes of the peripheral blood was not reduced in most patients. The percentage of target binding cells in patients was essentially the same as that in normal controls, and the activity of NK cells from normal donors was not affected by the coexistence of mononuclear cells obtained from the peripheral blood of patients. Furthermore, the alpha-IFN production in response to HeLa cells persistently infected with measles virus was reduced in all patients studied, except for one case with acquired idiopathic sideroblastic anaemia. The augmented activity of NK cells induced by alpha-IFN was variable, but remained at lower levels than the endogenous activity of NK cells in normal controls. These findings suggest that not only the intrinsic defect and reduced number of NK cells but also the dysfunction of the NK-IFN system may be responsible for the reduced activity of NK cells in patients with preleukaemic states.
The alterations of stimulus-induced membrane potential changes, superoxide (O2-)-producing capacity and phagocytic activity during differentiation of human granulocytes were investigated in the human leukemia cell lines HL-60 and KG-1 differentiating in vitro and in human leukemic granulocytes obtained from chronic myelogenous leukemia patients. HL-60 cells incubated with dimethyl sulfoxide or with retinoic acid showed progressively increasing O2- production as well as membrane potential changes (depolarization) on contact with phorbol myristate acetate or the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine, with a concomitant increase in the proportion of mature cells of the granulocytic type. Phagocytosis of latex particles, yeast, and oil droplets appeared 24 h after incubation with dimethyl sulfoxide and anteceded the increment of O2- production and membrane potential changes, both of which appeared concomitantly 3 d after incubation with dimethyl sulfoxide. Similar findings were observed when immature and mature granulocytes obtained from chronic myelogenous leukemia patients were stimulated by phorbol ester, the chemotactic peptide, or calcium ionophore A23187, and the amount of O2- production was parallel to the magnitude of membrane potential changes. HL-60 and KG-1 cells incubated for 1-6 d with phorbol myristate acetate showed neither O2- production nor membrane potential changes on contact with phorbol ester, chemotactic peptide, or A23187, although such cells resembled macrophages morphologically, and their phagocytic activity was significantly increased. O2- production and membrane potential changes in normal granulocytes induced by phorbol ester, chemotactic peptide and A23187 were inhibited by 2-deoxyglucose. These findings indicate that the O2--producing system and the system provoking membrane potential changes may develop concomitantly as human granulocytes mature and differentiate, and that the development of these systems and of phagocytic activity may be independently regulated.
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Five patients with multiple myeloma were treated with human lymphoblastoid interferon (HLBI). HLBI, 3 X 10(6) IU/day, was administered daily for more than two weeks by intramuscular injection. Out of four evaluable patients, a minor response was obtained in 3 patients. In these responders, one patient developed pleural effusion due to the infiltration of myeloma cells during the administration of HLBI, and drug resistance was observed in another patient during the re-administration of HLBI. Therefore, out of six evaluable courses, a minor response was obtained in 3 courses of HLBI treatment. No severe side effects were observed. Thrombocytopenia, general malaise, liver dysfunction and anorexia were the main reasons for discontinuation of HLBI administration. On the basis of the preliminary study, it is concluded that HLBI is worth trying in the management of refractory multiple myeloma.
The effect of DIDS, a specific inhibitor of anion transport in the erythrocyte membrane, on the ADP-stimulated aggregation of gel-filtered bovine blood platelets was examined. Marked inhibition of aggregation was observed at concentrations of more than 5 x 10(-5)M DIDS. On preincubation with platelets for 30 min, DIDS was more potent and significant inhibition was observed at concentrations of over 2 x 10(-7)M. Since ADP-stimulated aggregation of bovine gel-filtered platelets precedes the release reaction, these results suggest that an anion transport system in the plasma membrane is involved in platelet aggregation.
The reverse permeation of salicylate ion and the effect of bovine serum albumin on the permeation were studied in a sodium salicylate-sodium oxalate-water system. In passive transport the permeation flux of an ion is expressed by the linear combination of the two terms which represent the concentration and electric potential gradients. Because the mobility of the sodium ion is greater than the oxalate ion, salicylate ion moves against the concentration gradient, and follows the electric potential gradient in the initial stage of permeation. The reverse permeation of salicylate ion through a cellulose membrane was accelerated with a high concentration ratio of oxalate to salicylate ions and reached a maximum value after 10 hr in the absence of bovine serum albumin. After reaching a maximum value, the salicylate ion permeated along the concentration gradient. The maximum concentration efficiency was 11.2%. In the presence of bovine serum albumin, the reverse permeation of salicylate ion reached a maximum value after 3 hr.
Various chemical inducers have effects on the induction of terminal differentiation of human myelogenous leukemia cell lines. We studied morphological and functional changes of human leukemia cells freshly obtained from patients using 12-0-tetradecanoyl phorbol-13-acetate (TPA), retinoic acid (RA) or dimethyl sulphoxide (DMSO). The myeloid leukemia cells cultured with TPA became adherent to plastic culture dishes, and then developed macrophage-like morphology with long filamentous pseudopods within 48 h incubation. They showed marked enhancement of the ability to phagocytose latex particles. But these acquired properties did not always parallel each other, suggesting that the mechanism of functional maturation of leukemic cells induced by chemical agents was not identical with that of morphological changes. On the other hand, the lymphoid leukemia cells did not show morphological and functional changes when cultured with the above inducers. It is suggested that exposure of leukemic cells to TPA for relatively short times (12-24 h) may be useful for determining whether they are of myeloid or lymphoid origin. These characteristic changes were also observed in leukemic cells from the myeloid or lymphoid crisis of chronic myelogenous leukemia.
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