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Biomedical subjects

A Shirahata

Publications and source records attributed to A Shirahata.

At least 91 records · Page 5Linked to original sources
PubMed1995

Inhibition of cell growth by combination of alpha-difluoromethylornithine and an inhibitor of spermine synthase.

The inhibitory effect on cell growth of a combination of alpha-difluoromethylornithine (DFMO) and an inhibitor of aminopropyl transferase was examined. N-(3-aminopropyl)cyclohexylamine (APCHA) and trans-4-methylcyclohexylamine (4MCHA) were used as inhibitors of spermine and spermidine synthases, respectively. Combination of DFMO and APCHA showed strong inhibitory effects on the growth of FM3A cultured cells and P388 leukemia cells in mice, compared with DFMO alone. The prolongation of survival time of P388 leukemia-bearing mice by DFMO (1,500 mg/kg) was 1.12-fold, while that by DFMO (1,500 mg/kg) plus APCHA (25 mg/kg) was 1.30-fold. The prolongation of survival time nearly paralleled the decrease of P388 leukemia cells in mice. However, the antiproliferative effect of DFMO was not strengthened by 4MCHA in the above two experimental systems. In the FM3A cell culture system, both putrescine and spermidine contents were decreased by DFMO, but spermine content did not decrease significantly. When APCHA was added to the medium with DFMO, spermine content was decreased greatly but a compensatory increase in spermidine was observed. Spermidine content in P388 leukemia cells was also decreased by DFMO, the increase in spermine was suppressed but a compensatory increase in spermidine was observed. Nevertheless, the spermidine content remained significantly low compared with the value in non-treated P388 leukemia cells. Thus, the results indicate that the antiproliferative effect of DFMO was strengthened by APCHA due to the decrease in spermine content, and that the decrease in total amount of spermidine and spermine, especially the decrease in spermine, is necessary for inhibition of cell growth.

Animals↗
PubMed1995

Enzymatic aminopropylation of certain secondary amines.

Two unusual aminopropyl acceptors found in a survey of putrescine binding sites of mammalian spermidine synthase, N-methylputrescine (I) and 4-aminomethylpiperidine (II), were examined for their aminopropyl derivatives. Studies under in vitro incubation conditions suggested that the aminopropyl derivatives of the secondary amine of I and II, N4-methylspermidine (Is) and 1-N-(3-aminopropyl)-4-aminomethylpiperidine (IIs), and of the primary amine of I and II, N8-methylspermidine (Ip) and 4-[N-(3-aminopropyl)aminomethyl]piperidine (IIp), respectively, were biosynthesized by rat spermidine synthase. Studies on the cell culture system of cultured rat hepatoma (HTC) cells treated with alpha-difluoromethylornithine, an ornithine decarboxylase inhibitor, clearly showed the presence of Is and Ip when I was administered, and IIs and IIp when II was administered, with no detection of putrescine or spermidine. These results suggested that mammalian spermidine synthase can transfer the aminopropyl moiety of decarboxylated S-adenosylmethionine to certain secondary amines in living cells.

Animals↗
PubMed1995

Structural requirement for neurotrophic activity of spermine in cultured rat hippocampal neurons.

We investigated the structure-activity relationship for the neurotrophic activity of spermine by comparing the effects of several synthetic spermine analogues on the survival of cultured rat hippocampal neurons. N,N'-bis(3-aminopropyl)-1,6-hexanediamine and N,N'-bis(3-aminopropyl)-1,3-propanediamine did not promote the neuronal survival, suggesting that the central butanediamine structure is essential for the neurotrophic activity. Furthermore, N1,N12-bis(ethyl)spermine significantly promoted the neuron survival, but its maximum effect was smaller than that of spermine, indicating that two terminal primary amines are required for full agonist activity. The structural requirement for the neurotrophic activity of spermine was different from those for any other reported activities of polyamines.

Animals↗
PubMed1995

Antagonistic effect of N-(3-Aminopropyl)cyclohexylamine on neurotrophic action of spermine in primary cultured rat hippocampal and cerebellar neurons.

We previously found that spermine potently promotes the neuronal survival and regeneration of primary cultured brain neurons. N-(3-Aminopropyl)cyclohexylamine (APCHA) was originally developed as a spermine synthase inhibitor. To test if endogenous spermine biosynthesis contributes to neuronal survival and morphogenesis, we examined the effects of APCHA in primary cultured rat hippocampal and cerebellar neurons. APCHA at concentrations up to 10(-6) M did not affect the neuronal survival, but significantly blocked the survival-promoting effect of spermine (10(-8) M). APCHA also blocked the spermine-induced promotion of neurite regeneration following axotomy. Unlike APCHA, another cyclohexylamine derivative trans-4-methylcyclohexylamine did not affect the neurotrophic effect of spermine. These results suggest that in primary cultured brain neurons, APCHA works as a spermine antagonist rather than as a spermine synthesis inhibitor.

Animals↗
PubMed1995

Transforming growth factor beta 1 and beta 2 induce down-modulation of thrombomodulin in human umbilical vein endothelial cells.

To investigate the effects of transforming growth factor-betas (TGF-betas) on endothelial anticoagulant activity, we assayed thrombomodulin (TM) activity and antigen levels of human umbilical vein endothelial cells (HUVECs) incubated with TGF-betas in vitro. TGF-beta 1 suppressed surface TM activity and surface TM antigen levels maximally 12 h after incubation in dose-dependent manners. TGF-beta 2 was almost equipotent with TGF-beta 1 for the suppression of them. Both TGF-betas suppressed total TM antigen level in HUVECs, and the time course of the suppression was similar to that of the cell surface TM antigen level. The maximal reductions of TM mRNA levels by TGF-betas were observed at several hours ahead of those observed in both surface and total TM antigens levels, suggesting that the TGF-beta-mediated suppression of TM antigen of HUVECs is primarily regulated at the TM mRNA level. Our present work suggests that the down-modulation of TM level induced by TGF-betas in HUVECs contributes in vivo to promoting the thrombogenesis either at the sites of injury of vessel walls, such as atherosclerotic lesions where TGF-beta 1 is released from platelets, smooth muscle cells and monocytes, or at neovascular walls in tumors secreting TGF-beta 2.

Cells, Cultured↗
PubMed1995

[Hand-mirror cells acute lymphoblastic leukemia (L3)].

To our knowledge, this report represents the first confirmed case in Japan of a 15-year-old boy with acute lymphoblastic leukemia (ALL).L3 with hand-mirror cells (HMC) in the bone marrow. HMC lymphoid leukemia is an unusual variant of ALL in which the bone marrow lymphoblasts manifest distinctive hand-mirror morphologic features. HMC lymphoblast is characterized by an asymmetric foot-like cytoplasmic process that extends from the portion of the cell, thus giving it the light-microscopic appearance responsible for its name. Besides ALL, HMC has been reported in acute myeloblastic leukemia (AML), blastic crisis of chronic myelogenous leukemia, non-Hodgkin's lymphoma, and infectious mononucleosis. HMC has been reported to be prevalent in ALL.L1 and L2 as compared with L3.

Adolescent↗
PubMed1995

[Vitamin K].

Explore the source record for details and available documents.

Humans↗
PubMed1994

N-(3-aminopropyl)-cyclohexylamine blocks facilitation by spermidine of N-methyl-DL-aspartate-induced seizure in mice in vivo.

The facilitating or antagonizing effects of polyamine analogues on N-methyl-DL-aspartate (NMDLA)-induced seizures were investigated using mice. Intracerebroventricular injection of spermidine and spermine, but not putrescine, shortened the latency to appearance of clonic convulsion induced by subcutaneous administration of NMDLA. Injection of N-(3-aminopropyl)cyclohexylamine (APCHA) alone did not affect the NMDLA-induced seizure. However, APCHA, when administered together with spermidine, clearly antagonized the facilitating effect of spermidine on the NMDLA-induced seizure. Another cyclohexylamine derivative, trans-4-methylcyclohexylamine, did not block the effect of spermidine. APCHA also antagonized the facilitation by D-serine of NMDLA-induced seizure, although the blocking effect for D-serine was weaker than that for spermidine. APCHA should be useful as a new tool for pharmacological studies on the neuromodulatory action of polyamines.

Animals↗
PubMed1994

Correlation between the inhibition of cell growth by bis(ethyl)polyamine analogues and the decrease in the function of mitochondria.

The antiproliferating effect of nine kinds of bis(ethyl)polyamine analogues [three kinds each of bis(ethyl)triamine, bis(ethyl)tetraamine and bis(ethyl)pentaamine] was compared using FM3A cells. The inhibitory effect was in the order BE4444 > BE3443 > BE4334 > or = BE444 > BE343 > BE333 > BE44 > BE34 > BE33. Our results indicate that not only polyamine deficiency but also the accumulation of polyamine analogues is involved in the inhibition of cell growth. Accumulation of bis(ethyl)polyamine analogues caused the inhibition of protein synthesis and the decrease in the ATP content. The protein synthetic system in mitochondria was more strongly inhibited by bis(ethyl)polyamine analogues than that in the cytoplasm. Under conditions such that cytoplasmic protein synthesis was inhibited by 50% by bis(ethyl)polyamine analogues, mitochondrial protein synthesis was almost completely inhibited. Mitochondrial Ile-tRNA formation was inhibited by bis(ethyl)polyamine analogues at the concentrations that cytoplasmic Ile-tRNA formation was stimulated. This may be one of the reasons for the selective inhibition of mitochondrial protein synthesis. This inhibition was followed by the decrease in ATP content, swelling of mitochondria and depletion of mitochondrial DNA. These results suggest that the early event of metabolic change caused by bis(ethyl)polyamine analogues in cells is the inhibition of protein synthesis, especially of mitochondrial protein synthesis.

Adenosine Triphosphate↗
PubMed1993

Correlation between the inhibition of cell growth by accumulated polyamines and the decrease of magnesium and ATP.

The mechanism of the antiproliferation effect of spermidine and spermine was studied using a cell culture system of mouse FM3A cells. The addition of either 10 mM spermidine or 2 mM spermine to the growth medium containing 0.9 mM Mg2+ greatly inhibited cell growth (more than 90%). A decrease in the Mg2+ concentration to 50 microM in the growth medium, but without the polyamine addition, did not influence cell growth. However, the concentrations of spermidine and spermine necessary for the inhibition of cell growth when cells were cultured in the presence of 50 microM Mg2+ were much smaller (2 mM spermidine and 0.15 mM spermine). Nevertheless, the amount of polyamines accumulating in cells which could cause the inhibition of cell growth was almost the same, regardless of the large difference in the added polyamine concentrations. At the early stage of polyamine accumulation, the inhibition of cell growth correlated with the decrease of Mg2+ content, but not with a decrease of the ATP content. The decrease in Mg2+ content correlated well with the inhibition of macromolecular synthesis, especially protein synthesis. Thus, the inhibition of cell growth at the early stage of polyamine accumulation was thought to be due to the inactivation of ribosomes through the replacement of Mg2+ on magnesium-binding sites by polyamines. The decrease in Mg2+ content was mainly caused by the inhibition of Mg2+ transport by polyamines. At the later stage of polyamine accumulation, a decrease in ATP content was also observed. This was followed by swelling of the mitochondria, which may be a symptom of the subsequent cell death.

Adenosine Triphosphate↗
PubMed1993

Interaction of a polyamine analogue, 1,19-bis-(ethylamino)-5,10,15- triazanonadecane (BE-4-4-4-4), with DNA and effect on growth, survival, and polyamine levels in seven human brain tumor cell lines.

Computer graphics modeling and physicochemical studies of spermine-DNA interactions, as well as experiments in cell culture, indicate that a polyamine analogue with strong affinity for nucleic acids but poor ability to condense and aggregate DNA in vitro should act as an antiproliferative agent if it can enter cells. On the basis of our studies of polyamine-DNA interactions, we designed a pentamine, 1,19-bis(ethylamino)-5,10,15- triazanonadecane (BE-4-4-4-4), that had these characteristics. Measurement of melting temperature and ultraviolet light scattering studies show that the affinity of this analogue for calf-thymus DNA is about 4 times higher than that of spermine, whereas its ability to aggregate DNA is slightly poorer than that of spermine. Studies in U-87 MG, U-251 MG, SF-126, SF-188, SF-763, SF-767, and DAOY human brain tumor cells in tissue culture showed that treatment for more than 96 h with concentrations of 5 microM BE-4-4-4-4 or greater inhibited growth; decreased levels of putrescine, spermidine, and spermine; and decreased colony-forming ability in all cell lines. The cytotoxicity of the analogue varied among cell lines; DAOY and SF-767 were the most sensitive and the most resistant lines, respectively. In SF-763 cells, growth inhibition by BE-4-4-4-4 could be partially reversed by the addition of putrescine, spermidine, or spermine 1 day after BE-4-4-4-4 addition, but in U-251 MG cells, growth inhibition was reversed only by spermine and not by other polyamines. When any of the naturally occurring polyamines was added simultaneously with BE-4-4-4-4, growth inhibition was completely blocked. The data suggest that a threshold intracellular concentration of BE-4-4-4-4 is needed to manifest the growth-inhibitory and cytotoxic effects. In most cell lines, once that threshold level is reached, the growth-inhibitory and cytotoxic properties of the analogue are manifest irrespective of cellular polyamine levels. Further increases in the BE-4-4-4-4 concentration or incubation time reduce the intracellular polyamine levels but do not significantly increase growth inhibition. In U-87 MG and DAOY cells, however, prolonged incubation with higher concentrations of BE-4-4-4-4 causes additional growth inhibition along with depletion of intracellular polyamines.(ABSTRACT TRUNCATED AT 400 WORDS)

Brain Neoplasms↗
PubMed1993

Overproduction of S-adenosylmethionine decarboxylase in ethylglyoxal-bis(guanylhydrazone)-resistant mouse FM3A cells.

A variant cell line, termed SAM-1, which overproduced S-adenosylmethionine decarboxylase (AdoMetDC), was isolated by treatment of mouse FM3A cells with N-methyl-N'-nitro-N-nitrosoguanidine and subsequent incubation with ethylglyoxal bis(guanylhydrazone), an inhibitor of the enzyme. The cells were resistant to ethylglyoxal bis(guanylhydrazone), and showed AdoMetDC activity approximately five-times higher than control cells. The rate of AdoMetDC synthesis and the amount of AdoMetDC existing in SAM-1 cells were about five-times those in control cells. The amount of AdoMetDC mRNA existing in SAM-1 cells was five-times more than that in control cells. The amount of 5'-([(Z)-4-amino-2-butenyl]methylamino)-5'-deoxyadenosine, an irreversible inhibitor of AdoMetDC, necessary to inhibit cell growth was also five-times more in SAM-1 cells than in control cells. However, the following were the same in both SAM-1 and control cells; the amount of genomic DNA for AdoMetDC, the size and nucleotide sequence of 5' untranslated region of AdoMetDC mRNA, the deduced amino acid sequence (334 residues) from the nucleotide sequence of AdoMetDC cDNA and the degradation rate (t1/2 = about 4 h) of AdoMetDC. In addition, AdoMetDC mRNA in control cells was slightly more stable than that in SAM-1 cells. The results indicate that the overproduction of AdoMetDC in SAM-1 cells was caused by the increase of AdoMetDC mRNA. The variant cell line is convenient for studying the regulation of AdoMetDC and the physiological function of polyamines.

Adenosylmethionine Decarboxylase↗
PubMed1993

Effects of inhibitors of spermidine synthase and spermine synthase on polyamine synthesis in rat tissues.

Several inhibitors of aminopropyltransferases, developed recently in this laboratory, were tested for their specificity by measuring their effects on six enzyme activities related to polyamine biosynthesis and interconversion. Two of them, trans-4-methylcyclohexylamine (4MCHA) and N-(3-aminopropyl)cyclohexylamine (APCHA), selectively and potently inhibited the activities of spermidine synthase and spermine synthase, respectively. They were subjected to in vivo studies using rats. Oral administration of 4MCHA or APCHA dissolved in drinking water (0.02 and 0.1%) available ad lib. for a period of 10 days or 4 months caused a specific and marked decrease in spermidine or spermine in tissues (such as a 95% decrease) with a compensatory increase of spermine or spermidine, respectively, but without any observable change in the growth of the treated rats. Also, with extreme reduction of spermidine or spermine, when their sum was approximately constant, the activity of S-adenosyl-methionine decarboxylase in these tissues was enhanced significantly with no change in the activity of ornithine decarboxylase. These results suggested a separate role for spermidine or spermine in the in vivo enhancement of S-adenosylmethionine decarboxylase activity.

Adenosylmethionine Decarboxylase↗
PubMed1993

[New approach to the diagnosis of disseminated intravascular coagulation in childhood].

The peak incidence of DIC in the pediatric age group is in the neonatal period. The newborn infant is particularly susceptible to DIC because of several handicaps, such as physiological hypofunction of anticoagulant and fibrinolytic systems, an underdeveloped capacity in the reticuloendothelial system and a tendency to develop acidosis, hypothermia, hypoxia and shock. Although some criteria have been reported for the diagnosis of DIC in adults, based on clinical and laboratory findings, these are not necessarily applicable to the diagnosis of DIC in newborn infants. This is because a large blood sample is required, a long period of time is necessary for assay and difference in several coagulation and fibrinolysis factors exist between newborn infants and adults. We therefore established a criteria for the diagnosis of DIC in newborn infants, based on data obtained from newborn infants with DIC. Diagnostic procedures of many molecular markers of hemostasis have been developed from this. Some of them, such as FDP-D dimer are valuable for the diagnosis in children but others are not necessarily useful because of the susceptibility to the venipuncture technique. Our criteria for the diagnosis of DIC in newborn infants must be modified in the diagnosis of DIC in very low birth weight infants.

Age Factors↗
PubMed1993

[Clinical application of fat soluble vitamins to retinopathy of prematurity and intracranial hemorrhage].

More than ten years ago, vitamin E supplementation was proposed as a prophylaxies against a syndrome that might be caused by oxygen toxicity and/or membrane instability such as retinopathy of prematurity (ROP), bronchopulmonary dysplasia (BPD), and periventricular/intraventricular hemorrhage (PVH/IVH). The results obtained from several studies to date does not support the usefulness of vitamin E supplements in the prevention of BPD. However, several reports have suggested the utility of vitamin E in the prevention and/or inhibition of progress of ROP, although the use of vitamin E in premature infants for the possible prevention of severe ROP should still be considered investigational. On the other hand, the aggregate of evidence to date support the efficacy of tocopherol in preventing IVH of extremely premature infants. It is postulated that oxidative damage to capillary endothelial membranes of the subependymal layer predisposed the premature infant to subependymal bleeding and that vitamin E might act as an antioxidant to prevent such damage, thereby reducing the risk of hemorrhage.

Cerebral Hemorrhage↗
PubMed1993

[Vitamin K].

In this paper, highly sensitive methods for measuring of vitamin K in serum, feces and milk are described. Vitamin K1 and the menaquinone family from the serum, feces and milk were separated by high performance liquid chromatography. The separated K1 and menaquinone families were detected by a fluorescence spectrophotometer after their reduction with sodium borohydride or platinum catalyst column. In these methods, minimum detectable quantities of K1, menaquinone-4 and menaquinone-7 were 10, 10 and 50 pg, respectively. More than 95% of supplemented vitamin K1, menaquinone-4 and menaquinone-7 were recovered in the serum and feces. Furthermore, the method of Hirauchi et al. for measurement of vitamin K1 2,3-epoxide in human plasma is introduced.

Chromatography, High Pressure Liquid↗
PubMed1992

Mechanism of the inhibition of cell growth by N1,N12-bis(ethyl)spermine.

The mechanism of the antiproliferation effect of N1,N12-bis(ethyl)spermine (BESPM) was studied in detail using mouse FM3A cells, since this polyamine analogue mimics the functions of spermine in several aspects [Igarashi, K., Kashiwagi, K., Fukuchi, J., Isobe, Y., Otomo, S. & Shirahata, A. (1990) Biochem. Biophys. Res. Commun. 172, 715-720]. Our results indicate that not only the decrease in sperimine and spermine caused by BESPM but also its accumulation play important roles on the inhibition of cell growth by BESPM, since BESPM accumulated in cells at a concentration fivefold that of spermidine in control cells. In comparison with the polaymine-deficient cells caused by alpha-difluoromethylornithine, an inhibitor of ornithine decarboxylase, and ethylglyoxal bis(guanylhydrazone), an inhibitor of S-adenosylmethionine decarboxylase, the behavior of polyamine-deficient cells caused by BESPM was different as follows: the inhibition of cell growth by BESPM was not abrogated by spermine or spermidine; polyamine uptake, which is stimulated during polyamine deficiency, was greatly inhibited, while spermidine/spermine N1-acetyltransferase activity, which is inhibited during polyamine deficiency, was enhanced in BESPM-treated cells; thymidine kinase activity did not decrease in BESPM-treated cells; inhibition of cell growth and macromolecule synthesis by BESPM correlated with the swelling of mitochondria and the decrease in ATP content; BESPM caused cell death when incubated together for several days. The role of BESPM accumulation on inhibition of cell growth is discussed.

Adenosine Triphosphate↗