[Sinus pause, sinus arrest].
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Electrocardiography↗
Biomedical subjects
T Wakatsuki
Publications and source records attributed to T Wakatsuki.
Efficacy of coronary angioplasty with a cutting balloon in a case of repeated coronary stenosis.
A 59-year-old man with a 90% stenosis of the left anterior descending coronary artery was treated with percutaneous transluminal coronary angioplasty (PTCA) using a conventional balloon. The stenosis recurred repeatedly and was retreated by PTCA three times. The stenosis again recurred, so coronary angioplasty using a cutting balloon was performed. During the present admission, we planned to place a coronary stent in the LAD to prevent restenosis. Anticoagulation is necessary after coronary stenting. However, anticoagulation was contraindicated in this patient because of ulcerative colitis. Therefore, coronary angioplasty using a cutting balloon was performed instead. The stenosis decreased from 90% to 25%, but a coronary dissection occurred. An angiogram performed 3 months later showed no stenosis, with resolution of the coronary dissection. The use of a cutting balloon preventing coronary artery restenosis following balloon angioplasty is a promising new technique which warrants further study.
Aortic Dissection↗
Increased growth of brown adipose tissue but its reduced thermogenic activity in creatine-depleted rats fed beta-guanidinopropionic acid.
To study the responses of thermogenic activity in brown adipose tissue (BAT) to creatine depletion, male Wistar rats were fed creatine analogue beta-guanidinopropionic acid (beta-GPA) for about 10 weeks. Compared to control rats, a marked decrease in the levels of high-energy phosphates, such as phosphocreatine and ATP, was noted in BAT of beta-GPA rats. Conversely, upward trends in other chemical components (DNA, glycogen, and total protein) in BAT as well as an increase in BAT mass were observed in beta-GPA rats, suggesting a tendency to hyperplasia of the BAT. The thermogenic activity (which was assessed by guanosine 5'-diphosphate binding to BAT mitochondria) in the mitochondria recovered from BAT of beta-GPA rats, however, was not increased in response to such changes but rather decreased. Moreover, uncoupling protein (UCP) content in the mitochondrial fraction of beta-GPA rats was significantly lower than that in control rats (the relative amounts were 77 +/- 6 and 100 +/- 4%, respectively). Nevertheless, surprisingly, the level of UCP mRNA was remarkably greater in beta-GPA rats than in control rats. These observations indicate that there is a discordance between BAT growth and activity in beta-GPA rats, thereby suggesting that a failure on and after UCP translation may be involved in the impairment of BAT thermogenic activity with creatine depletion. The impairment of BAT thermogenic activity, that is, UCP activity may indicate that uncoupling or heat production was inhibited in order to increase the ATP synthesis in BAT of beta-GPA rats in compensation for a reduction in the levels of high-energy phosphates (including ATP), with resultant hypothermia.
Adipose Tissue, Brown↗
Metal oxidoreduction by microbial cells.
For many organisms, some heavy metals in external media are essential at low concentrations but are toxic at high concentrations. Strongly toxic heavy metals are toxic even at low concentrations. Recently, it was proven that changes of valencies of Fe, Cu and Mn were necessary for these metals to be utilized by organisms, especially microorganism. The valencies of Hg and Cr are changed by reducing systems of cells in the process of detoxifying them. Thus, the processes of oxidoreduction of these metals are important for biological systems of metal-autoregulation and metal-mediated regulation. Metal ion-specific reducing enzyme systems function in the cell surface layer of microorganisms. These enzymes require NADH or NADPH as an electron donor and FMN or FAD as an electron carrier component. Electron transport may be operated by transplamsa-membrane redox systems. Metal ion reductases are also found in the cytoplasm. The affinities of metal ions to ligand residues change with the valence of the metal elements and mutual interactions of various metal ions are important for regulation of oxidoreduction states. Microorganisms can utilize essential metal elements and detoxify excess metals by respective reducing enzyme systems and by regulating movement of heavy metal ions.
Bacteria↗
Responses of brown adipose tissue activity to unloading in rats.
To study the responses of thermogenic activity in brown adipose tissue (BAT) to unloading, male Wistar rats were hindlimb suspended for 10 days. Compared with control rats, a significant increase in the BAT-to-body mass ratio and considerable differences in chemical components in BAT were observed in the hindlimb-suspended rats. These findings indicate a marked increase in the thermogenic capacity in BAT of the experimental group. Likewise, the thermogenic activity (which was assessed by guanosine 5'-diphosphate binding to BAT mitochondria) was markedly greater in the mitochondria recovered from BAT of the hindlimb-suspended rats than in those from the control rats (1,610 +/- 450 vs. 202 +/- 132 pmol recovered). Moreover, the uncoupling protein content in the BAT mitochondrial fraction of the hindlimb-suspended rats was significantly higher (1.6-fold) than that in the control rats. As was expected, the uncoupling protein mRNA expression was greater in hindlimb-suspended rats than in control animals. These results suggest that chronic hindlimb suspension leads to an increase in both the thermogenic capacity and the activity in BAT of rats.
Adipose Tissue, Brown↗
Changes of contractile properties of extensor digitorum longus in response to creatine-analogue administration and/or hindlimb suspension in rats.
Changes of contractile properties of extensor digitorum longus in response to hindlimb suspension and/or altered high-energy phosphate contents were studied in rats. A reduction of high-energy phosphates, especially phosphocreatine, was seen in rats fed creatine analogue beta-guanidinopropionic acid (beta-GPA), but they were elevated after 10-d supplementation of creatine. The one-half relaxation time was increased by feeding beta-GPA, but was normalized by creatine supply. The fatigue resistance of creatine-depleted muscle was significantly improved, but tended to decrease by suspension and creatine supply, although it was still better than that in the control diet group. It is indicated that the contractile properties of muscle are influenced by the high-energy phosphate content. It is also suggested that the endurance capacity may be influenced by the mitochondrial respiratory capacity, but not necessarily by the levels of high-energy phosphates.
Animals↗
A distinct mRNA encoding a soluble form of ICAM-1 molecule expressed in human tissues.
A soluble form of ICAM-1 (sICAM-1) have been observed in normal human serum (Rothlein et al., J. Immunol. 147, 3788-3793) and at elevated levels in inflammatory and tumor bearing status (Seth et al., Lancet, 338, 83-84; Giavazzi et al., Canc. Res. 52, 2628-2630; Harning et al., Canc. Res., 51, 5003-5005). However, the mechanism to produce the sICAM-1 has been still unknown. In this report we presented evidence for the presence of the mRNA specifically encoding sICAM-1, which is probably generated by alternative splice donor site selection. A 19-base deletion occurred right upstream of the transmembrane region gave rise to reading frameshift and eliminate the entire transmembrane and cytoplasmic domains, resulting in incapability of ICAM-1 molecules to reside in the membrane. A reverse transcription-polymerase chain reaction (RT-PCR) using a primer pair specific to sICAM-1 revealed a positive expression in all tissues analyzed, though the amount and the ratio to the conventional species varied slightly from tissue to tissue. Inflammatory cytokines displayed a complex pattern in the ICAM-1 mRNA expression depending on the combination of cytokines and the cultured cell lines used.
Alternative Splicing↗
A variant mRNA species encoding a truncated form of Fas antigen in the rat liver.
A variant form of the Fas antigen cDNA was isolated from the rat liver. The cDNA contains an insertion sequence of 25 nucleotide residues and codes for a protein consisting of 65 amino acids representing a signal peptide and an N-terminal portion of the extracytoplasmic domain of Fas antigen. The expression of a variant mRNA species is less abundant in the liver compared with that of an ordinary mRNA species. The variant mRNA, though its function is unknown, is plausibly produced by an alternative splicing.
Amino Acid Sequence↗
Metabolic adaptation of skeletal muscles to gravitational unloading.
Responses of high-energy phosphates and metabolic properties to hindlimb suspension were studied in adult rats. The relative content of phosphocreatine (PCr) in the calf muscles was significantly higher in rats suspended for 10 days than in age-matched cage controls. The Pi/PCr ratio, where Pi is inorganic phosphate, in suspended muscles was less than controls. The absolute weights of soleus and medial gastrocnemius (MG) were approximately 40% less than controls. Although the % fiber distribution in MG was unchanged, the % slow fibers decreased and the % fibers which were classified as both slow and fast was increased in soleus. The activities (per unit weight or protein) of succinate dehydrogenase and lactate dehydrogenase in soleus were unchanged but those of cytochrome oxidase, beta-hydroxyacyl CoA dehydrogenase, and citrate synthase were decreased following unloading. None of these enzyme activities in MG changed. However, the total levels of all enzymes in whole muscles decreased by suspension. It is suggested that shift of slow muscle toward fast type by unloading is associated with a decrease in mitochondrial biogenesis. Further, gravitational unloading affected the levels of muscle proteins differently even in the same mitochondrial enzymes.
Animals↗
Responses of beta-adrenoceptor in rat soleus to phosphorus compound levels and/or unloading.
Responses of beta-adrenoceptor (beta-AR) in rat soleus to gravitational unloading and/or changes in the levels of phosphorus compounds by feeding either creatine or its analogue beta-guanidinopropionic acid (beta-GPA) were studied. A decrease in the density of beta-AR (about -35%) was induced by 10 days of hindlimb suspension, but the affinity of the receptor was unaffected. Suspension unloading tended to increase the levels of adenosine triphosphate and phosphocreatine and decrease inorganic phosphate. Even without unloading, the beta-AR density decreased after an oral creatine supplementation (about -20%), which also tended to elevate the high-energy phosphate levels in muscle. However, an elevation of beta-AR density was induced (about +36%) after chronic depletion of high-energy phosphates by feeding beta-GPA (about +125%). Data suggest that the density of beta-AR in muscle is elevated if the high-energy phosphate contents are chronically decreased and vice versa. However, it may not be directly related to the degree of muscle contractile activity.
Animals↗
Effects of vesnarinone (OPC-8212) on Ca(2+)-activated K channels and cytosolic Ca2+ in cultured smooth muscle cells from porcine coronary artery.
Vesnarinone is a new, non vasodilating cardiotonic agent. This study compared the effects of vesnarinone and amrinone, a phosphodiesterase (PDE) inhibitors with vasodilating actions, on cultured smooth muscle cells from the porcine coronary artery. Application of vesnarinone (10(-4) M) or amrinone (10(-4) M) to the bath solution in cell-attached patches activated the KCa channel having a conductance of 133 pS (bath 2.7 mM K, pipette 140 mM K). Application of vesnarinone to the cytosolic side had no direct effect on KCa channel activities in inside-out patches. Activation of the KCa channel was suppressed when the intracellular production of cAMP was suppressed by preincubation with carbachol (10(-6) M). Amrinone, but not vesnarinone, lowered [Ca2+]i in the K(+)-depolarized smooth muscle cells (K+ = 70 mM). These results suggest that vesnarinone exerts an additional effect on [Ca2+]i that is independent of PDE inhibition. The difference in the effects on [Ca2+]i in vascular smooth muscle cells may explain in part the differing actions of these agents on vascular relaxation.
Amrinone↗
Responses of contractile properties in rat soleus to high-energy phosphates and/or unloading.
Responses of contractile properties of soleus to unloading and/or changes in high-energy phosphate contents were studied in rats. Reduction of high-energy phosphates, especially phosphocreatine, in ankle extensors was induced by feeding beta-guanidinopropionic acid (beta-GPA). The major finding in the study was that the fatigability and speed-related contractile properties responded to unloading and creatine supplementation in a similar manner. The high-energy phosphate contents tended to be elevated after 10-d supplementation of creatine and hindlimb suspension. The shift toward slow-type, mainly due to an increased one-half relaxation time, was seen in rats fed beta-GPA. Such a shift was reversed by feeding creatine or by hindlimb suspension; however, the suspension-induced shift of contractile properties toward fast-type was not prevented completely by beta-GPA feeding. Although the muscle fatigue resistance did not change by beta-GPA feeding alone, the decrease in fatigue resistance following suspension and creatine supply was less in the beta-GPA group. It is suggested that the levels of high-energy phosphates and tension production play important roles in the regulation of contractile properties of the soleus muscle.
Animals↗
Non-insulin and non-exercise related increase of glucose utilization in rats and mice.
The effects of high-energy phosphate contents in muscles on glucose tolerance and glucose uptake into tissues were studied in rats and mice. Enhanced glucose tolerance associated with depleted high-energy phosphates and elevated glycogen content in muscles and liver was observed in animals fed creatine analogue beta-guanidinopropionic acid (beta-GPA). Distribution of infused 2-[1-14C]deoxy-D-glucose in tissues especially in the soleus muscle, kidney, and brain was greater in mice fed beta-GPA than controls. The glucose uptake was decreased when the contents of ATP and glycogen were normalized following creatine supplementation. Plasma insulin in animals at rest was lower and its concentration after intraperitoneal glucose infusion tended to be less in animals fed beta-GPA than controls (p > 0.05), although the pattern of insulin response to glucose loading was similar to the control. The daily voluntary activity in beta-GPA fed mice was also less than controls. These results suggest that improved glucose tolerance is not related to elevated insulin concentration and/or decreased glycogen following exercise. Such improvement may be due to an increased mitochondrial energy metabolism caused by depletion of high-energy phosphates.
Animals↗
[A rapid, simple and inexpensive preparation method of small amount, but large numbers of E. coli plasmids].
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Cloning, Molecular↗
Regulation of hsp70 induction in thermotolerant HeLa cells.
Upon exposure to heat shock, non-thermotolerant (NT) HeLa cells transiently synthesize a large amount of 70-kDa heat-shock protein (hsp70), whereas thermotolerant (TT) cells synthesize a small amount of hsp70. When the hsp70 mRNA of HeLa cells was analyzed, it became apparent that hsp70 mRNA in TT cells did not increase following heat shock, whereas hsp70 mRNA in NT cells did increase dramatically. A further analysis of the activation of the heat-shock transcription factor (HSF) showed that significant activation of HSF was observed immediately after heat shock in both NT and TT cells. However, activated HSF was rapidly repressed in the TT cells, but not in the NT cells. Thus, the decreased induction of hsp70 synthesis observed in the TT HeLa cells may be due to the immediate repression of activated cellular HSF, which probably results in the reduced induction of hsp70 mRNA. The hsp70 content in the TT cells was usually higher than in the NT cells. However, after heat-shock treatment, the hsp70 content of the NT cells increased to nearly the level of the TT cells concomitant with the repression of hsp70 synthesis. The association of activated HSF with hsp70 was observed in both NT and TT cells, and the amount of HSF-hsp70 complex within the cell increased in proportion to the increase in hsp70 in the cells. These findings strongly suggest that the activity of HSF is negatively regulated by the intracellular content of hsp70 in these cells. Furthermore, in vitro experiments on the activation of HSF suggest that HSFs of NT and TT cells may have different properties, or an unknown factor may exist which regulates HSF activation in these cells.
Base Sequence↗
Regulation of hsp70 synthesis induced by cupric sulfate and zinc sulfate in thermotolerant HeLa cells.
Upon exposure to the heavy metals copper and zinc, a large amount of 70,000-Da heat shock protein (hsp70) was synthesized in normal, non-thermotolerant (NT) HeLa cells, whereas only a little increase of hsp70 synthesis was observed in thermotolerant (TT) cells. To determine the inhibition mechanism of hsp70 induction in the TT cells, we first analyzed the hsp70 mRNA of these cells. Hsp70 mRNA in the NT cells increased immediately after exposure to these metals. In TT cells, however, the increase of hsp70 mRNA was delayed, even though it eventually increased to a similar or slightly lower level compared with NT cells. Further analysis of the activation of heat shock transcription factor (HSF) showed that a significant activation of HSF was observed immediately after exposure to these metals in the NT cells, whereas the activation of HSF was initially repressed in the TT cells. Thus, the decreased induction of hsp70 synthesis observed in the TT cells seemed to be due to the reduced translation of hsp70 mRNA and also in part to the reduced activation of HSF. Furthermore, by the gel mobility shift assay using anti-hsp70 antibody, the association of copper- and zinc-activated HSF with hsp70 was observed in both NT and TT cells. The amount of HSF-hsp70 complex was prominent in TT cells, in which the hsp70 content was 5 to 10 times higher than that in the NT cells. These findings strongly suggest that the activity of HSF is negatively regulated by hsp70 in the TT cells.
Base Sequence↗
Effects of culture temperature on the expression of heat-shock proteins in murine ts85 cells.
The murine temperature-sensitive cell-cycle mutant, ts85, shows an abnormal induction of heat-shock proteins which is different from the wild type FM3A cells. This paper explores the effect of culture temperature on the expression of heat shock proteins in ts85 cells. When ts85 cells were maintained at 33 or 37 degrees C, these cells synthesized heat-shock protein (hsp) 70 following continuous heating at 39 degrees C or subsequent incubation after heating at 42 degrees C for 15 min. In contrast, these conditions are not conducive for hsp70 synthesis by FM3A cells. Moreover, ts85 cells which were maintained at 37 degrees C synthesized hsp70 following continuous heating at 42 degrees C or subsequent incubation after heating at 45 degrees C for 15 min. The synthesis of hsp70 in these cells corresponded to an increased level of hsp70 mRNA. Furthermore, the constitutive hsp105 level of cells maintained at 33 degrees C was only half of that of cells which were maintained at 37 degrees C, and cells maintained at 33 degrees C were more sensitive to subsequent heat treatment than those maintained at 37 degrees C. These results indicate that culture temperature not only affects the induction of hsp70 mRNA, but also cellular levels of hsp105 and the resulting thermal sensitivity of ts85 cells. These findings suggest that the other phenotypic characteristic of the mutant ts85 cells is also affected by culture temperature.
Animals↗
Characteristic induction of 70,000 da-heat shock protein and metallothionein by zinc in HeLa cells.
The synthesis of a 70,000 dalton-heat shock protein (hsp70) is one of several heat shock proteins induced in HeLa cells during the incubation in medium containing zinc sulphate. The synthesis of hsp70 was increased in the presence of 200 microM zinc sulphate and above, but not at 100 microM zinc sulphate. On the other hand, the synthesis of metallothionein was activated in the presence of 100 microM zinc sulphate and above. Uptake of zinc into the cells depended on the concentration of zinc sulphate in the medium. The separation of intracellular zinc into three fractions by gel filtration chromatography; high molecular, metallothionein, and low molecular fractions, showed that zinc in the low molecular weight and metallothionein fractions was elevated in the presence of 100 microM zinc sulphate in the medium, whereas increase in the zinc content of the high molecular weight fraction occurred at 200 microM zinc sulphate and above. Inhibition of cell growth and cellular protein synthesis was also observed at 200 microM zinc sulphate and above, but not at 100 microM. From these findings, since the induction of hsp70 synthesis and inhibition of cell growth occurred concomitantly with the increase of zinc in the high and low molecular weight fractions, hsp70 seemed not to function in the detoxification of zinc, but it may participate in the repair of zinc-induced damage.
Cell Division↗