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In the first half, this paper reviews the so-called “Plasma Gene Transfection” and “Plasma Molecular Introduction” which are the technology to introduce molecules such as plasmid DNA, proteins, and dye molecules into cells by discharge plasma treatment. In the latter half, the authors’ method with the micro discharge plasma is reviewed. This method induces the cell’s spontaneous uptake of external molecules by endocytosis which is triggered by the complex of weak stimuli produced by the micro discharge plasma treatment less than 10 ms and enables high introduction efficiency and high cell viability simultaneously. There is almost no damage to the cells and external introducing molecules, and there is almost no unintended integration of the introduced gene into the chromosome. The authors would like to call this feature “Random Genome Integration-Free.” This feature would enable safe and clean gene and molecular introduction for practical applications such as medicine and breeding.

The gene/molecule introduction method using surface discharge enables transfection into a wider area, compared with the micro-plasma method. In this research, as the first step in elucidating the gene transfer mechanism in the surface discharge method, it was revealed that endocytosis is the main introduction mechanism in the surface discharge method, and that 80% of the introduction is due to endocytosis. Furthermore, the relationship between the size of the target molecule and the transfection efficiency was investigated. As a result, it was made clear that when target molecules with a molecular weight of up to 10 000 bp, the transfection efficiency is proportional to the collision frequency between the target molecule and cell. On the other hand, it became clear that when the molecular weight exceeds 10 000 bp, the transfection efficiency decreases with increasing molecular weight, and that almost no introduction is made at 15 000 bp or more.

Heated scallop-shell (HSS) nano-particles, prepared using a wet grinding mill, and microparticles were examined for their antibacterial activity against vegetative bacterial cells and spores. The median diameters of the nano-particles and micro-particles were approximately 20 nm and 30 µm, respectively. The antibacterial activity of HSS against Escherichia coli increased with an increase in concentration, regardless of particle size; however, the antibacterial activity of the nano-particles was much higher than that of micro-particles. The sporicidal activity of the nano-particles was also much higher than that of micro-particles, with HSS nano-particles able to kill Bacillus subtilis spores. A reduction of more than three orders of magnitude for B. subtilis spores was confirmed following a 30 min treatment at 5 mg/ml and 60℃, showing that the combination of HSS nano-particle treatment with mild heating was particularly effective for controlling bacterial spores.

Selective peroxisome proliferator-activated receptor gamma (PPARγ) modulators are expected to be a novel class of drugs improving plasma glucose levels without PPARγ-related adverse effects. As a continuation of our studies for (-)-Cercosporamide derivatives as selective PPARγ modulators, we synthesized substituted naphthalene type compounds and identified the most potent compound 15 (EC(50) = 0.94 nM, E(max) = 38%). Compound 15 selectively activated PPARγ transcription and did not activate PPARα and PPARδ. The potassium salt of compound 15 showed a high solubility and a good oral bioavailability (58%). Oral administration of the potassium salt remarkably improved the plasma glucose levels of female Zucker diabetic fatty rats at 1 mg/kg. Moreover, it did not cause a plasma volume increase or a cardiac enlargement in Wistar-Imamichi rats, even at 100 mg/kg.

Peroxisome proliferator-activated receptor gamma (PPARγ) is a potential drug target for treating type 2 diabetes. The selective PPARγ modulators (SPPARMs), which partially activate the PPARγ transcriptional activity, are considered to improve the plasma glucose level with attenuated PPARγ related adverse effects. However, the relationships between desired pharmacological profiles and ligand specific PPARγ transcriptional profiles have been unclear. And there is also little knowledge of how to control ligand specific PPARγ transcriptional profiles. Herein, we present synthesis of novel derivatives containing substituent at naphthalene C3 position of compound 1. The novel derivatives showed various maximal efficacies as PPARγ partial agonist.

The effects of strength-mismatch between the base and weld metal in welded joints on CTOD and Charpy impact toughness of heat-affected zone (HAZ) are investigated in this study. Both toughness properties are compared, in consideration of the effect of the local brittle zone (LBZ) in HAZ, and strength-mismatch effects are discussed analytically. Two types of welded joints for 490 MPa strength class structural steel with the same thickness of 25 mm, which were strength-matched and overmatched joints, are prepared under the same welded conditions (heat input = 4 KJ/mm) with different welding consumables. The critical CTOD of the HAZ for the overmatched joint exhibits the lower values in wide temperature range of ductile-to-brittle fracture transition than that for the matched joint, whereas microstructures and total LBZ size along crack-tip are consistent with each other. On the contrary, significant difference between the matched and overmatched joints in the lower temperature range cannot be observed in the Charpy impact toughness. The reason why the strength-mismatch effects on the fracture toughness are different between the CTOD and Charpy impact tests is discussed.

A sphingosine-1-phosphate receptor 1 (S1P1) antagonist is expected to be an anti-angiogenic compound; however, there are few reports that demonstrated that a S1P1 inhibitor improved the disease state in an angiogenic animal model. Since we determined that a prototype S1P1 antagonist was an in vivo angiogenesis inhibitor, we developed the derivatives to acquire more effective compounds. In this report, we show the S1P1 antagonistic activity of some representatives, especially compound 5 {sodium 4-[(4-butoxyphenyl)thio]-2'-[{4-[(heptylthio)methyl]-2-hydroxyphenyl}(hydroxy)methyl]biphenyl-3-sulfonate}. The IC50 values calculated from an intracellular cyclic AMP measurement assay and a [33P]sphingosine-1-phosphate (Sph-1-P)/S1P1 binding assay were 38 and 200 nM, respectively. A subtype specificity test for the other Sph-1-P receptors showed that compound 5 was the S1P1-directional antagonist. It also inhibited the proliferation, migration, and tube formation of human umbilical vein endothelial cells stimulated by Sph-1-P with the IC50 values of 18, 650, and 230 nM, respectively. A cytotoxicity assay concurrently performed with a tube formation assay supported the hypothesis that these biological effects were not due to its cytotoxicity. Furthermore, administration (10 mg/kg, intravenously) to anesthetized Sprague-Dawley rats inhibited Sph-1-P-induced hypotension by 100-90% for 30 min. This is presumably through the inhibition of Sph-1-P-induced vasorelaxation, mainly by the blocking of S1P1 and/or S1P3. Taken together, these results show that compound 5 is an inhibitor of in vitro and in vivo Sph-1-P signaling, and that it will be useful to elucidate the in vivo effect of Sph-1-P on vascular endothelial cells.

This study was conducted to investigate whether or not a food substitute (Dr. BAANs(R)) containing three bioactive components L-arginine, omega-3 polyunsaturated fatty acid, and ribonucleic acid, supplied orally to 15 overweight patients, may have efficacy to prevent or improve the metabolic syndrome of these patients. To provide supporting data for this clinical study, the in vivo fatty acid metabolism of obese mice was analyzed using (125)I labeled 15-(p-iodophenyl)-9-methylpentadecanoic acid (9MPA) in the tissues' lipid pool. After 3 months of intervention, the results showed that there were improvements observed in liver functions, lipid profiles and metabolic syndrome marker. Significant differences were also found in the values of blood pressure, body weight, percentage of body fat, and body mass index. In the animal study, the tissue uptake of (125)I-9MPA at 10 min after injection was higher in obese mice than in the control mice and the treatment with Dr. BAANs(R) in obese mice decreased the uptake significantly. The final product metabolite of p-iodophenylacetic acid in obese mice was increased significantly by the treatment. In conclusion, this food substitute may have a beneficial effect for the prevention or improvement of metabolic syndrome.

Aptamers are an attractive candidate for a molecular-targeted drug since they can show ability to precisely recognize proteins with a large affinity similar to antibodies. A drawback of aptamers is...

We examined the effect of trimidox-induced apoptosis involvement of p53 in the NALM-6 cell line of acute lymphoblastic leukemia. Trimidox has been shown to increase the induction of p53. Phosphorylation of p53 protein at Ser-15 and Ser-20 residues was activated earlier than the obvious increase in p53 expression. Pifithrin-α, a p53 inhibitor, significantly prevented trimidox-induced apoptotic characteristics, as detected by nuclear morphological observation and DNA fragmentation. Trimidox-induced apoptosis was enhanced or attenuated by transfection with wild-type or dominant-negative p53 containing expression vectors, respectively. These results indicate that one of the induction mechanisms of apoptosis by trimidox is the mediated augmentation of p53. Keywords:: trimidox, apoptosis, p53

A series of Sodium 4-[(4-butoxyphenyl)thio]-2'-substituted-1,1'-biphenyl-3- sulfonates were identified as functional sphingosine-1-phosphate (S1P) antagonists with selectivity for the S1P(1) receptor subtype starting from chemical lead 2, which was found while screening our in-house compound library. We performed chemical modifications on each regional structure of compound 2, for example, on the three ring compartments, the benzyl substituents, and the long alkyl chain part. The introduction of a biphenyl skeletal structure and the installation of a hydroxyl group onto the terminal carbon in the side-chain region resulted in the potent derivative 35c, which showed >500-fold more potent S1P(1) inhibitory activity than lead compound 2. We report herein the synthesis and structure-activity relationships of structurally novel S1P(1) receptor antagonists.

Intrathecal (i.t.) administration into mice of S -(+)-fenfluramine (0.01–0.1 nmol), a serotonin (5-hydroxytryptamine, 5-HT) releaser, produced a behavioral response consisting of scratching, biting and licking. Here, we report the behavioral characteristics and the involvement of interactions between 5-HT and substance P (SP) systems in the S -(+)-fenfluramine-induced behavioral response. The S -(+)-fenfluramine-induced behavioral response peaked at 5–15 min and almost disappeared at 20 min after injection. The behavior induced by S -(+)-fenfluramine (0.1 nmol) was dose-dependently inhibited by an intraperitoneal injection of morphine (0.02–0.5 mg/kg), suggesting that the behavioral response is related to nociception. The S -(+)-fenfluramine-induced nociceptive behavior was significantly inhibited by pretreatment with 5-HT antiserum and co-administration of ketanserin, a selective 5-HT2 receptor antagonist. However, WAY-100635, a selective 5-HT1A receptor antagonist, and ramosetron, a selective 5-HT3 receptor antagonist, were not active. On the other hand, SP antiserum and RP67580, a selective neurokinin-1 (NK1) receptor antagonist, significantly inhibited S -(+)-fenfluramine-induced nociceptive behavior. These results suggest that i.t.-administered S -(+)-fenfluramine releases SP through the activation of 5-HT2 receptors subsequent to 5-HT release, and, as a result, produces nociceptive behavior.

The engineering method for fracture toughness evaluation of beam-to-column weld HAZ using one-bead welds, called one-bead weld method, has been examined in this study. Materials used are 26 structural steels of 400 to 490 MPa strength class produced by different milling conditions. One-bead welds are made by CO2 arc welding with single bevel groove under heat inputs of 4.0 to 5.0 kJ/mm. From these welded joints, the Charpy specimens and 3-point bend CTOD specimens with the same thickness of 10mm are extracted. The notch is located at the weld HAZ on a straight bond side perpendicular to the plate surface. Charpy tests and CTOD tests are carried out at 0°C and -40°C, respectively, where the CTOD test temperature is determined on the basis of WES2808 correlation formula. It has been found that the Charpy absorbed energy is affected by the chemical composition of the base steel, such as C, Mn, P, S, N, and Ti. Based on similar test results, an engineering parameter, fHAZ, is proposed by the Building Center of Japan for estimating the HAZ toughness. However, the present study indicates that the total amount of N is not a proper measure, but the free N controls the HAZ toughness. From the fracture mechanical point of view, the correlation between the Charpy energy and the critical CTOD is useful. It is noted that the CTOD test results are sensitive to the length of CGHAZ along the crack front. The Charpy-CTOD correlation with respect to the HAZ toughness should be made with attention to the base-metal chemical compositions as well as the CGHAZ size along the crack front.

This study has been done to know what kind of factors in plasmas and processes on cells induce plasma gene transfection. We evaluated the contribution weight of three groups of the effects and processes, i.e. electrical, chemical and biochemical ones, inducing gene transfection. First, the laser produced plasma (LPP) was employed to estimate the contribution of the chemical factors. Second, liposomes were fabricated and employed to evaluate the effects of plasma irradiation on membrane under the condition without biochemical reaction. Third, the clathrin-dependent endocytosis, one of the biochemical processes was suppressed. It becomes clear that chemical factors (radicals and reactive oxygen/nitrogen species) do not work by itself alone and electrical factors (electrical current, charge and field) are essential to plasma gene transfection. It turned out the clathrin-dependent endocytosis is the process of the transfection against the 60% in all the transfected cells. The endocytosis and electrical poration are dominant in plasma gene transfection, and neither permeation through ion channels nor chemical poration is dominant processes. The simultaneous achievement of high transfection efficiency and high cell survivability is attributed to the optimization of the contribution weight among three groups of processes by controlling the weight of electrical and chemical factors.

Intrathecal (i.t.) administration into mice of N-ethylmaleimide (NEM), a cysteine protease inhibitor, produced a characteristic behavioral response, the biting and/or licking of the hindpaw and the tail along with slight hindlimb scratching directed toward the flank. The behavior induced by NEM was inhibited by the intraperitoneal injection of morphine. We have recently reported that dynorphin A and, more potently big dynorphin, consisting of dynorphins A and B, produce the same type of nociceptive response whereas dynorphin B does not [Tan-No K, Esashi A, Nakagawasai O, Niijima F, Tadano T, Sakurada C, Sakurada T, Bakalkin G, Terenius L, Kisara K. Intrathecally administered big dynorphin, a prodynorphin-derived peptide, produces nociceptive behavior through an N-methyl-d-aspartate receptor mechanism. Brain Res 2002;952:7-14]. The NEM-induced nociceptive behavior was inhibited by pretreatment with dynorphin A- or dynorphin B-antiserum and each antiserum also reduced the nociceptive effects of i.t.-injected synthetic big dynorphin. The characteristic NEM-evoked response was not observed in prodynorphin knockout mice. Naloxone, an opioid receptor antagonist, had no effects on the NEM-induced behavior. Ifenprodil, arcaine and agmatine, antagonists at the polyamine recognition site on the N-methyl-D-aspartate (NMDA) receptor ion-channel complex, and MK-801, an NMDA ion-channel blocker inhibited the NEM-induced effects. Ro25-6981, an antagonist of the NMDA receptor subtype containing NR2B subunit was not active. NEM completely inhibited degradation of dynorphin A by soluble and particulate fractions of mouse spinal cord. Collectively, the results demonstrate that endogenous prodynorphin-derived peptides are pronociceptive in uninjured animals, and required for the NEM-induced behavior. The NEM effects may be mediated through inhibition of the degradation of endogenous dynorphins, presumably big dynorphin that in turn activates the NMDA receptor ion-channel complex by acting on the polyamine recognition site.

The inhibitory effect of repetitiously administered loperamide, a peripheral mu-opioid receptor agonist and well-recognized antidiarrheal agent, on mouse gastrointestinal transit was compared with that of morphine in order to examine the development of tolerance to mu-opioid receptor agonist-induced constipation (antitransit effect). When administered subcutaneously 15 min before the oral injection of charcoal meal, loperamide (0.1-30 mg/kg) and morphine (1-8 mg/kg) dose-dependently and significantly inhibited gastrointestinal transit of charcoal with the ID(50) values of 1.6 (0.3-7.1) mg/kg and 3.6 (1.5-8.5) mg/kg, respectively. When loperamide (30 mg/kg) was administered twice daily for 2 days, the antitransit effect was significantly reduced. On the other hand, morphine did not develop tolerance in even more severe conditions than those of loperamide. It is known that P-glycoprotein, an ATP-dependent drug efflux pump, is involved in the development of tolerance to morphine analgesia. The tolerance observed with loperamide was significantly prevented by cyclosporin (30 mg/kg, i.p.), a P-glycoprotein inhibitor, thus the ID(50) value in loperamide-tolerant mice was markedly reduced from >1000 mg/kg to 40 (2.7-603.0) mg/kg by cyclosporin. These results indicate that loperamide, different from morphine, readily develops tolerance to the inhibitory effect on mouse gastrointestinal transit, and the P-glycoprotein may be involved in the development of tolerance to the antitransit effect of loperamide.

A series of our studies have shown that formation of cholesterol-supersaturated bile in patients with cholesterol gallstone disease is causatively related to decreased gallbladder contractility and mucin hypersecretion by the gallbladder. Supersaturated bile may modify the composition of gallbladder membranes so that the transduction of smooth muscle regulatory signals is impaired, and it may enhance the inflammation-induced mucin secretion by the gallbladder. To achieve a better understanding of the mechanism by which supersaturated bile impairs the contractility, we studied changes in the expression levels of gallbladder cholecystokinin (CCK-A) receptor messenger ribonucleic acid (mRNA) in prairie dogs fed a high-cholesterol diet. Levels of pathobiological determinants in arachidonate metabolism which are important for mucin secretion were also measured in their bile. Adult male prairie dogs were randomly assigned to receive either a semisynthetic diet (SSD) or an SSD plus 1.2% cholesterol (a high-cholesterol diet) for 2-, 4-, and 6-week periods. The contractile force in response to CCK-octapeptide (CCK-8) was measured by using gallbladder muscle strips. The mRNA levels of the CCK-A receptor were determined by reverse-transcription polymerase chain reaction (RT-PCR). Parallel to the increase in the cholesterol saturation index, the contractile responses to CCK-8 decreased in the animals fed a high-cholesterol diet for 4 weeks and markedly decreased in the animals with gallstone formation. However, in contrast to the decreased contractility, the steady-state mRNA levels of the gallbladder CCK-A receptor were significantly increased in the animals fed a high-cholesterol diet in comparison with the corresponding control animals. In the bile, a high-cholesterol diet caused an increase in the proportion of arachidonyl-phosphatidylcholine species, where phospholipase A(2) activity, prostaglandin E(2), and mucin concentrations were increased parallel to the feeding period. Up-regulation of the CCK-A receptor mRNA in the gallbladder of animals fed a high-cholesterol diet associated with decreased contractility may be due to an impairment of CCK signaling related to increased membrane cholesterol contents and its related reaction of biological compensation in order to increase the receptor concentration. The results of the present study suggest that in prairie dogs fed a high-cholesterol diet both a decrease in gallbladder contractility related to impairment of CCK signaling and phospholipase A(2) (PLA(2))-induced mucosal inflammation in the gallbladder with associated biliary alterations favoring cholesterol crystal formation pathogenetically contribute to the formation of cholesterol gallstones.

The highly sensitive and precise determination method for trace amounts of Sb and B in high-purity iron and steel has been established by the isotope dilution/Inductively coupled plasma mass spectrometry. For the determination of Sb, the iron matrix was separated by anion-exchange chromatography using Dowex I-X8 in hydrofluoric acid solution, and the isotope ratio ( 121 Sb/ 123 Sb) of the HNO 3 /H 2 O 2 eluate was measured by ICP-MS. The isobaric interference of 123 Te was corrected by subtracting the intensity of 123 Te obtained by the relative intensity of 123 Te and 125 Te. For the determination of B, after the treatment of sulfuric acid-phosphoric acid fuming for the complete decomposition of boron nitride, B was separated by the anion-exchange chromatography using Amberlite IRA-743 at pH 8. The CyDTA was added to prevent the hydrolysis of iron. The isotope ratio ( 11 B/ 10 B) of the HCI eluate was measured by ICP-MS. By these methods, Sb and B in the range of μg/g to sub-μg/g could be determined with good precision. The limit of detection is 5.8 ng/g for Sb and 16 ng/g for B in steel.

大腸脂肪腫は比較的稀な粘膜下腫瘍であるが径が大きくなるにつれ,腸重積を合併して発見される頻度が高い.今回われわれは術前にCT,内視鏡検査にて脂肪腫の診断を得て腹腔鏡補助下手術を選択できた症例を経験したので報告する.症例は77歳,男性.左下腹部痛にて近医を受診,注腸造影にて下行結腸下端に表面平滑な円形腫瘤とcoiled spring signを認め,腸重積の診断にて当院紹介となる. CTにて下行結腸内腔に径約3cmの内部が-93HDと脂肪組織に近い濃度のCT値を示す円形の腫瘤を認めた.内視鏡検査ではcushionsignを示す柔らかな粘膜下腫瘍の所見を示しており大腸脂肪腫と診断,腹腔鏡補助下に結腸部分切除を行った.病理組織検査では5×2.5×2.7cm大の成熟した脂肪腫で,漿膜下より発生し腸管内腔に向かって亜有茎性,気球状の発育を示していた.巨大な大腸脂肪腫で術前に質的診断が得られた症例は侵襲の少ない腹腔鏡補助下手術の良い適応であると考えられる.

Microstructure change in high-purity irons was investigated at various cooling rates after single pass hot-rolling. Large columnar grains, which are named mono-layer honeycomb grains, developed through the thickness when high-purity iron (C:1.5 mass ppm, N:0.8 mass ppm, S:1.5 mass ppm, Si + Mn + P < 1.0 mass ppm) was hot-rolled at 1273 K and cooled in a furnace with the average cooling rate of about 8.5 × 10 -3 K/s. Mono-layer honeycomb grains developed in neither high-purity iron water-quenched after hot-rolling, nor middle-purity (C: 1.5 mass ppm, N: 9 mass ppm, S: 10 mass ppm, Si+Mn+P: 42 mass ppm) and low-purity (C: 20 mass ppm, N: 8 mass ppm, S: 10 mass ppm. Si +Mn+ P: 270 mass ppm) irons. The crystal orientation of mono-layer honeycomb grains was determined by electron back scattering pattern method. The crystal orientation of mono-layer honeycomb grains does not have any specific α-texture. Grain growth behavior was investigated by using the cold-rolled and annealed irons. The grains in the high-purity iron grew faster and larger than those in middle-purity and low-purity irons. It is considered that the hot-rolled γ-grains in the high-purity iron recrystallize and coarsen in cooling, and then the γ-grains transform into α-grains. The α-grains grow larger due to the effect of purification. Consequently, mono-layer honeycomb grains are developed in high-purity iron.

We have developed a new micro-discharge plasma (MDP)-based gene transfection method, which transfers genes into cells with high efficiency and low cytotoxicity; however, the mechanism underlying the method is still unknown. Studies revealed that the N-acetylcysteine-mediated inhibition of reactive oxygen species (ROS) activity completely abolished gene transfer. In this study, we used laser-produced plasma to demonstrate that gene transfer does not occur in the absence of electrical factors. Our results show that both electrical and chemical factors are necessary for gene transfer inside cells by microplasma irradiation. This indicates that plasma-mediated gene transfection utilizes the synergy between electrical and chemical factors. The electric field threshold required for transfection was approximately 1 kV m−1 in our MDP system. This indicates that MDP irradiation supplies sufficient concentrations of ROS, and the stimulation intensity of the electric field determines the transfection efficiency in our system. Gene transfer by plasma irradiation depends mainly on endocytosis, which accounts for at least 80% of the transfer, and clathrin-mediated endocytosis is a dominant endocytosis. In plasma-mediated gene transfection, alterations in electrical and chemical factors can independently regulate plasmid DNA adhesion and triggering of endocytosis, respectively. This implies that plasma characteristics can be adjusted according to target cell requirements, and the transfection process can be optimized with minimum damage to cells and maximum efficiency. This may explain how MDP simultaneously achieves high transfection efficiency with minimal cell damage.

Acquired drug resistance is a major factor in the failure of doxorubicin-based cancer chemotherapy. We determined the ability of cinchonine to reverse doxorubicin drug resistance in a doxorubicin-resistant leukaemia cell line (mouse P388/DOX). A non-cytotoxic concentration of cinchonine (10 μM) increased the sensitivity to doxorubicin of multidrug-resistant P388/DOX cells and significantly enhanced the doxorubicin-induced apoptosis and DNA fragmentation in resistant cells, but had no effect in parent cells. Time-course studies demonstrated that DNA fragmentation was present 24 h after incubation with doxorubicin and cinchonine, indicating that DNA degradation was a preceding event. In cultured cells, cinchonine increased the intracellular accumulation of doxorubicin in the resistant cells in a dose-dependent manner. Using flow cytometry to measure the inhibition of the P-glycoprotein (P-gp) dependent efflux of rhodamine 123, cinchonine was found to be considerably more effective than quinine. The results with cinchonine suggest that there may be quinine derivatives with a similar capacity to inhibit drug transport by P-gp. Additionally, the G2/M phase cell population in resistant cells is increased by doxorubicin/cinchonine treatment. Exposure of resistant cells to 1 μM doxorubicin and 10 μM cinchonine resulted in the expression of Fas (APO-1/CD95) in cells after 6 h. These studies demonstrate that the cell killing effects of doxorubicin and cinchonine in resistant cells are mediated, at least in part, by the induction of apoptosis.

Anesthetized open-chest dogs were subjected to 15-min myocardial ischemia followed by 2-h reperfusion to induce myocardial stunning. A novel Na(+)/H(+) exchange inhibitor 6,7,8,9-tetrahydro-2-methyl-5H-cyclohepta[b]pyridine-3-carbonylguanidine maleate (TY-12533), administered 10 min before or 10 min after start of ischemia (3 mg/kg/10 min, i.v.), did not affect reductions in regional myocardial wall thickening, blood flow and pH during ischemia, but it significantly improved recovery of the wall thickening and blood flow after reperfusion. These results indicate that TY-12533, even when administered during ischemia, could prevent myocardial stunning without affecting myocardial dysfunction or acidosis induced by brief ischemia.

We elucidated the interaction of small-conductance Ca2+-activated K+(SKCa) channels and L-type Ca2+channels in muscarinic receptor-mediated control of catecholamine secretion in the isolated perfused rat adrenal gland. The muscarinic agonist methacholine (10–300 μM) produced concentration-dependent increases in adrenal output of epinephrine and norepinephrine. The SKCachannel blocker apamin (1 μM) enhanced the methacholine-induced catecholamine responses. The facilitatory effect of apamin on the methacholine-induced catecholamine responses was not observed during treatment with the L-type Ca2+channel blocker nifedipine (3 μM) or Ca2+-free solution. Nifedipine did not affect the methacholine-induced catecholamine responses, but it inhibited the responses during treatment with apamin. The L-type Ca2+channel activator Bay k 8644 (1 μM) enhanced the methacholine-induced catecholamine responses, whereas the enhancement of the methacholine-induced epinephrine and norepinephrine responses were prevented and attenuated by apamin, respectively. These results suggest that SKCachannels are activated by muscarinic receptor stimulation, which inhibits the opening of L-type Ca2+channels and thereby attenuates adrenal catecholamine secretion.

To investigate the effect of angiotensin (ANG) II on aldosterone (ALDO) secretion, we measured arterial and adrenal venous plasma aldosterone concentrations in anesthetized dogs. The intraadrenal arterial infusion of ANG II (0.3 ng/kg/min) or potassium chloride (KCl) (0.6 mg/min) increased ALDO secretion. The changes in ALDO secretion in response to ANG II were tested during the concomitant arterial infusion of two graded doses of losartan (10 and 100 ng/kg/min), PD 123319 (50 and 500 ng/kg/min), nifedipine (25 and 250 ng/kg/min), or TMB-8 (2 and 20 microg/kg/min). All of these test drugs except PD123319 inhibited the ANG II-induced increase in ALDO secretion. Losartan did not affect the KCl-induced increase in ALDO secretion. These results indicate that ANG II acts on ANG II type 1 receptors in the adrenal gland and enhances ALDO secretion. They also suggest the involvement of both intracellular and extracellular calcium in the aldosterone response to stimulation by ANG II. Under these in vivo experimental conditions, the KCl-stimulated ALDO secretion does not appear to involve ANG II formation in the adrenal gland.

We examined the participation of endothelin ET A and ET B receptors in modulation by endothelin-1 of adrenal catecholamine secretion during cholinergic activation in pentobarbital-anesthetized dogs. Drugs were infused intra-arterially into the adrenal gland. Splanchnic nerve stimulation (1 and 3 Hz) increased adrenal catecholamine output in a frequency-dependent manner. Endothelin-1 (0.2, 0.6, and 2 ng/kg/min) enhanced the catecholamine response induced by the 3-Hz nerve stimulation. Under pretreatment with an endothelin ET A receptor antagonist ( R )-2-[( R )-2-[( S )-2-[[1-(hexahydro-1 H -azepinyl)]carbonyl]amino-4-methylpentanoyl]amino-3-(2-pyridyl) propionic acid (FR139317) (1 μg/kg/min), endothelin-1 suppressed the 1- and 3- Hz nerve stimulation-induced catecholamine response in a dose-dependent manner. No inhibitory or facilitatory effect of endothelin-1 was observed under simultaneous pretreatment with FR139317 and an endothelin ET B receptor antagonist N - cis 2,6-dimethylpiperidinocarbonyl- l -γ-methylleucyl- d -1-methoxycarbonyltryptophanyl- d -norleucine (BQ-788) (1 μg/kg/min) or under pretreatment with BQ-788 alone. These results suggest that in the dog adrenal gland, endothelin-1 facilitates and inhibits adrenal catecholamine secretion during cholinergic activation by stimulating endothelin ET A and ET B receptors, respectively.

The microstructure change of three kinds of high-purity irons (HP, MP and LP irons) by hot-rolling at various temperatures was investigated. Columnar grains in high-purity irons develop along the normal direction after hot-rolling in the γ-phase region. The growth of columnar grains depends on the purity of the iron: the width and height of columnar grains increase with the increase of the purity of the iron. Giant columnar grains of HP-iron grow to the center of the thickness. Columnar grains do not develop by hot-rolling in the α-phase region nor by heating in γ-phase region without hot-rolling. Apparently, the γ → a transformation after hot-rolling in the γ-phase region plays an important role in the growth of giant columnar grains. The crystal orientation of giant columnar grains was determined by an electron back scattering pattem method. The transformed α-texture was calculated from the y-texture with {112} by using the Krudjumov-Sachs relationship. The calculated α-texture is in good agreement with the measured crystal orientations except for {100} . Furthermore, the formation mechanism of giant columnar grains has been proposed.

We evaluated the effects of an adenylate cyclase activator, N, N-dimetyl-beta-alanine[3R-(3alpha,4alphabeta,5beta+ ++,6beta,6aalpha, 10alpha,10abeta,10balpha)]-5(acetyloxy)-3-eth enyldodecahydro-10, 10b-dihydroxy-3,4a,7,7,10a-pentamethyl-1-oxo-1H-naphtho[2, 1-b]pyran-6-yl ester hydrochloride (NKH477), on neural control of renal functions in anesthetized dogs. Renal nerve stimulation (2 Hz) increased renal norepinephrine efflux and reduced renal blood flow, glomerular filtration rate, urine flow rate, urinary Na(+) excretion and fractional Na(+) excretion. Intrarenal arterial infusion of NKH477 (300 ng/kg/min) suppressed the stimulation-induced reductions in renal blood flow and glomerular filtration rate and attenuated the reductions in urine flow rate and urinary Na(+) excretion but not the changes in renal norepinephrine efflux and fractional Na(+) excretion. Infusion of NKH477 did not affect the urinary responses induced by renal nerve stimulation at a lower frequency (0.5-1 Hz) which had little influence on renal blood flow and glomerular filtration rate. The present results demonstrate that NKH477 inhibits renal vasoconstriction and hypofiltration but not the enhanced tubular Na(+) reabsorption during activation of the renal sympathetic nervous system.

We examined effects of gamma-aminobutyric acid (GABA) on vasoconstriction and noradrenaline (NA) release induced by electrical renal nerve stimulation (RNS) in the isolated pump-perfused rat kidney. RNS (1 and 2 Hz for 2.5 min each, 0.5-ms duration, supramaximal voltage) increased renal perfusion pressure (PP) and renal NA efflux. GABA (3, 10 and 100 microM) attenuated the RNS-induced increases in PP by 10-40% (P

Although the renal nitric oxide (NO)-cyclic guanosine monophosphate (cGMP) system plays an important role in maintaining urinary sodium and water excretion, effects of an authentic NO donor sodium nitroprusside (SNP) on urine formation have been controversial. In this study, we examined whether SNP increases renal NO release and cGMP production and induces natriuresis in the denervated kidney of anesthetized dogs. The intrarenal arterial infusion of SNP at 10, 30, and 100 ng/kg/min did not affect renal function or NO-cGMP production. The higher dose of SNP (1.000 ng/kg/min) reduced systemic blood pressure and urine flow rate. The antidiuresis was observed also in the contralateral control kidney, the degree of which was larger than that observed in the ipsilateral SNP-infused kidney. During the SNP infusion, reductions in urinary Na + excretion, fractional Na + excretion. and urinary nitrite + nitrate excretion occurred in the control kidney but not in the SNP-infused kidney. Urinary cGMP excretion and renal venous plasma cGMP concentration were significantly increased during the SNP infusion in the SNP-infused kidney but not in the control kidney. These renal effects of SNP were similar to those obtained by intrarenal arterial infusion of a specific NO donor, NOC 7 (300 ng/kg/min). These results suggest that SNP can produce nitric oxide and increase cGMP levels in the kidney and suppress sodium reabsorption. but the natriuretic property of SNP may be masked by its counteracting effects including the systemic hypotension in anesthetized dogs.

C-glycosylated aryl tins have been prepared as versatile building blocks of physiologically stable glycomimetics for glycoepitopes that have been recognized to serve biologically important roles in cell biochemistry.

C-glycosylated biphenyls were prepared by the palladium-mediated cross-coupling reaction of C-glycosylated aryl tins with variously substituted aryl bromides, which will provide physiologically stable glycomimetics of various glycoepitopes. A C-sialylated biphenyl, a glycomimetic of biologically significant sialosides, is also available by this method.

IL-1 and TNF-alpha are known to be pleiotropic cytokines associated with various inflammatory conditions such as small intestinal injury after ischemia-reperfusion. FR167653 has been characterized as a potent suppressant of IL-1 and TNF-alpha production. The effect of FR167653 on intestinal reperfusion injury was investigated in a warm ischemia model of the canine gut. Sixteen mongrel dogs were divided into two groups: a control group and a FR group to which FR167653 was administered. Both the superior mesenteric artery and vein were clamped for 2 hr. Arterial pH, hepatic venous hemoglobin oxygen saturation, intramucosal pH, and the survival rate were well maintained in the FR group in comparison with the control group after reperfusion. FR167653 inhibited the expression of IL-1beta mRNA. Histologically, ischemia-reperfusion injury was more severe in the control group than the FR group. This study suggests that FR167653 inhibits proinflammatory cytokines and ameliorates ischemia-reperfusion injury of the small intestine.

C-glycosylated diphenylmethanes and C-glycosylated benzophenones were prepared by the palladium-catalyzed cross-coupling reaction of C-glycosylated aryl tins with various benzyl bromides or acid chlorides to provide physiologically stable glycomimetics of miscellaneous glycoepitopes.

In the course of the search for physiologically stable, structurally simple, and low molecular weight sLeX mimetics, aryl C-glycosides with carboxylic acid functionality 2 were found to be extremely potent inhibitors against L- and P-selectins with IC50 in the low microM range.

SUMMARY 1. We examined whether zaprinast, a putative cGMP-specific phosphodiesterase inhibitor, affects neural control of renal function in pentobarbital-anaesthetized dogs. 2. Renal nerve stimulation (1Hz, 1ms duration) reduced urine flow rate, urinary Na+ excretion (UNaV) and fractional excretion of Na+ (FENa) with little change in either renal blood flow (RBF) or glomerular filtration rate (GFR). 3. Intrarenal arterial infusion of zaprinast (10 and 100 μg/kg per min) increased basal urine flow rate, UNaV and FENa but not RBF or GFR. Zaprinast infusion (100 μg/kg per min) also increased renal venous plasma cGMP concentration and urinary cGMP excretion. 4. Renal nerve stimulation-induced reductions in UNaV and FENa were attenuated during zaprinast infusion, whereas the reduction in urine flow rate was resistant to zaprinast. 5. Renal nerve stimulation increased the renal venous plasma noradrenaline concentration and renal noradrenaline efflux, which remained unaffected during infusion of zaprinast (100 μg/kg per min). 6. The results of the present study suggest that zaprinast induces natriuresis and counteracts adrenergically induced anti-natriuresis by acting on renal tubular sites in the dog kidney in vivo.

We investigated the effect of cilnidipine, a novel dihydropyridine Ca2+ channel blocker possessing blocking actions on N-type and L-type voltage-dependent Ca2+ channels (VDCCs), in comparison with the L-type VDCC blocker nifedipine, on adrenal catecholamine secretion in response to splanchnic nerve stimulation (SNS), acetylcholine (ACh), the nicotinic receptor stimulant 1,1-dimethyl-4-phenyl-piperazinium (DMPP), and muscarine in anesthetized dogs. Ca2+ channel blockers and cholinergic agonists were infused and injected, respectively, into the adrenal gland through the phrenicoabdominal artery. Cilnidipine (0.3-3 microg/min) inhibited increases in both epinephrine (EPI) and norepinephrine (NE) output induced by SNS (2 Hz), ACh (1.5 microg), and DMPP (0.2 microg). However, cilnidipine inhibited increase in NE output induced by muscarine (1 microg) without affecting increase in EPI output. Nifedipine (0.3-3 microg/min) inhibited the ACh- and DMPP-induced increases in EPI and NE output without affecting the SNS- and muscarine-induced increases in EPI and NE output. From these results, it seems likely that the inhibition by cilnidipine of the SNS-induced EPI and NE secretion and of the muscarine-induced NE secretion is related to its blocking action on N-type VDCCs.