Kim, Yong-An;Yang, Seong-Hun;Lee, Seok-Hyeong;Lee, Gyeong-U;Park, Jong-Wan
Korean Journal of Materials Research
/
v.9
no.6
/
pp.583-588
/
1999
The characteristics of copper thin films and pattern filling capability were investigated by ECD. Prior to deposition of copper film, seed-Cu/Ta(TaN)/$SIO_2$(BPSG)/Si structure was manufactured. Copper deposition was performed with various current waveforms(DC/PC, 1~10,000Hz) and current densities(10~60 mA/$\textrm{cm}^2$) after pretreatment (Oxident removal, wetting) of seed-layer. Conformal pattern filling was performed using PC method with fast deposition rate of 6,000~8,000$\AA$/min. Heat-treated($450^{\circ}C$, 30min) copper films showed good resistivities of 1.8~2.1$\mu$$\Omega$.cm. According to the XRD analysis, (111)-preferred orientation of copper film was found in ECD-Cu/seed-Cu/Ta/$Sio_2$/Si structure. Also, we have successfully achieved to fill via holes with 0.35$\mu\textrm{m}$ width and 4:1 aspect ratio.
This study aimed at observing the effect of diazepam on the contractility of trachealis muscle isolated from canine trachea, possible involvement of central or peripheral type benzodiazepine receptor, and the calcium related mechanism of action of diazepam. Trachealis muscle strips of 15 mm long were suspended in an isolated organ bath containing 1 ml of physiologic salt solution maintained at $37^{\circ}C$, and aerated with 95% $O_2$ /5% $CO_2$. Isometric myography was performed. Diazepam reduced the basal tone concentration dependently, and this inhibitory action was not affected by neither flumazenil, a central benzodiazepine receptor antagonist, nor PK11195, a peripheral benzodiazepine receptor antagonist. Pretreatment with diazepam showed the inhibitory effect on the concentration-response curves to agonists such as bethanechol, 5-hydroxytryptamine and histamine. Diazepam also caused concentration-related inhibition of contraction with potassium chloride 30 mM. The effect of diazepam on the basal tone and potassium chloride-induced contraction with calcium channel blockers were compared. Similar results were obtained in canine trachealis with verapamil, nifedipine and diltiazem. These results suggest that diazepam relax an airway muscle not via specific receptors but by a similar action as calcium channel blockers in canine trachealis muscle.
In order to induce somatic embryogenesis from the stem explants and anther of Kalanchoe daigremontiana, the explants were cultured on MS medium supplemented with auxin (2,4-D, IAA, NAA) and/or cytokinin (BAP) for 8 weeks. Callus from explants was induced most efficiently on MS medium containing. 2.0mg/L NAA and 0.2mg/L BAP. Somatic embryogenesis in stem callus was formed by transfering embryogenic callus from induction media containing growth regulators to medium without growth regulators and then to the medium containing auxin and cytokinin (0.1 mg/L IAA and 1.0mg/L BAP). Callus formation occurred actively in the anthers at early uninucleate stage, and by low temperature pretreatment at $4^{\circ}C$ for 3days. Somatic embryogenesis from the anther callus was induced on MS medium containing 1.0mg/L NAA and 1.0mg/L BAP, 2.0mg/L NAA and 0.2mg/L BAP. The tetraploid of 5.4% was obtained among plants regenerated from anthers.
Photocurable sulfonated polyimide (SPI) polyelectrolyte containing chalcone group was prepared and fabricated on an alumina electrode pretreated with chalcone-containing silane-coupling agent. SPI films with bis(tetramethyl)ammonium 2,2'-benzidinedisulfonate ($Me_4N$-BDS)/4,4'-diaminochalcone (DAC)/pyromellitic dianhydride (PA)= 90/10/100 possessed very linear response(Y = -0.04528X+7.69446, $R^2=0.99675$) and showed resistance changing from 4.48 to $2.1k{\Omega}$ between 20 and 95 %RH. The response time for absorption and desorption measurements between 33 and 94 %RH% was about 79 s, which affirmed the high efficiency of crosslinked SPI film for rapid detection of humidity. A negative temperature coefficient showing $-0.49%RH/^{\circ}C$ was found and proper temperature compensation should be considered in future applications. Moreover, pretreatment of the substrates with chalcone-containing silane-coupling agent was performed to improve the water durability and the stability of the humidity sensors at a high humidity and a high temperature and long-term stability for 480 h. The crosslinked SPI films anchored to electrode substrate could be a promising material for the fabrication of efficient humidity sensors with superior characteristics compared to the commercially available sensors.
The contractile mechanisms of serotonin were investigated in the renal artery of a rabbit. The helical strips of isolated renal artery were immersed in the normal or $Ca^{2+}$-free tris-buffered Tyrode's solution, which was equilibrated with 100% $O_{2}$ at $35^{\circ}C$. The contraction by serotonin or norepinephrine (NE) began at $1{\times}10^{-7}\;M$ and reached the maximal contraction at $1{\times}10^{-5}\;M$. The maximal contraction by serotonin corresponded to $58.1{\pm}4.2%$ of maximal contraction by NE. Cyproheptadine, a serotonin receptor blocker, shifted the concentration-response curve to the right without any reduction in the maximum response but shifted that of NE to the right with reduction in maximum response. And phentolamine, an ${\alpha}-receptor$ blocker, shifted the concentration-response curve of serotonin or NE without any reduction in maximum responses. The $pA_{2}$ values for cyproheptadine against serotonin and NE were $10.35{\pm}0.04$ and $8.45{\pm}0.13$, respectively. The $pA_{2}$ values for phentolamine against serotonin and NE were $6.87{\pm}0.04$ and $8.14{\pm}0.08$, respectively. after the pretreatment with 6-hydroxydopamine, the contraction induced by 100 mM $K^{+}$, tyramine and serotonin reduced to $83.0{\pm}2.0$, $26.8{\pm}6.2$ and $82.0{\pm}3.5%$ of control, respectively. The contraction by serotonin in the $Ca^{2+}$-free Tyrode's solution was increased and sustained with the addition of $Ca^{2+}$ extracellulary. The serotonin-sensitive intracellular $Ca^{2+}$ pool was depleted completely by the pretreatment with NE, but the NE-sensitive intracellular $Ca^{2+}$ pool was depleted partially by the pretreatment with serotonin. From the above results, it is suggested that the contraction induced by serotonin in the renal artery of a rabbit may be due to mechanisms in which serotonin acts directly on specific serotonin receptors and also acts indirectly on ${\alpha}-adrenoceptors$ by displacing NE from neuronal stores.
This paper describes the development of $^{99m}Tc$ tricarbonyl cysteine as potential renal function diagnostic radiopharmaceutical and evaluation of its biological characteristics using experimental animals. l-Cysteine was labeled efficiently with $^{99m}Tc$ tricarbonyl precursor $([^{99m}Tc(CO)_3(H_2O)_3)]^{+})$ under 30 min heating at ${75^{\circ}C}$. Labeling yield and stability were analyzed by high performance liquid chromatography (HPLC). The biodistribution property of $^{99m}Tc$ tricarbonyl cysteine in mice and its dynamic imaging profiles in rabbits were carried out. To investigate the excretion mechanism of $^{99m}Tc$ tricarbonyl cysteine, tubular transport inhibition test with probenecid was adopted. $^{99m}Tc$ tricarbonyl cysteine was obtained with a high labeling yield under the moderate condition. The results of biodistribution experiments of $^{99m}Tc$ tricarbonyl cysteine in ICR mice at 3 and 90 min provided that $^{99m}Tc$ tricarbonyl cysteine was very highly accumulated in the kidney and bladder, thereby almost 99% of $^{99m}Tc$ tricarbonyl cysteine was excreted within 90 min post injection. The same results were confirmed by the whole body dynamic images for 30 minutes and static images in rabbits at given time intervals after injection. Renogram of $^{99m}Tc$ tricarbonyl cysteine in rabbits showed that its $T_{max}$ and $T_{1/2}$ of $^{99m}Tc$ tricarbonyl cysteine were $2.33{\pm}0.56$ and $4.30{\pm}0.79$ min, respectively. The $T_{max}$ of $^{99m}Tc$ tricarbonyl cysteine with probenecid pretreatment was $2.30{\pm}0.17$ min, whereas $T_{1/2}$ of that with probenecid pretreatment was $17.0{\pm}32.47$ min. $T_{1/2}$ of $^{99m}Tc$ tricarbonyl cysteine with probenecid pretreatment was significantly different, as compared to the result without probenecid (p<0.0001). The results showed that the excretion of $^{99m}Tc$ tricarbonyl cysteine was extremely affected by probenecid. Therefore, $^{99m}Tc$ tricarbonyl cysteine was rapidly excreted from the kidney principally by the tubular secretion.
Enzyme substitution with recombinant phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) is currently being explored for treatment of phenylketonuria (PKU), an autosomal recessive genetic disorder with mutations of the gene encoding phenylalanine-4-hydroxylase (EC 1.14.16.1). However, oral administration of PAL is limited because of proteolytic digestion in the gastrointestinal tract. The aim of this study was to determine the biochemical properties of PAL and delinate the susceptibility of wild-type PAL to pancreatic proteolysis by exploring several mutants, and to develop therapeutic drugs with PAL for PKU. The specific activity of PAL was assayed and its optimal pH, temperature stability, and intestinal protease susceptibility were investigated. Its $V_{max}$ values for phenylalanine and tyrosine were 1.77 and $0.47{\mu}mol$/ min/mg protein, respectively, and its $K_m$ values were $4.77{\times}10^{-4}$ and $4.37{\times}10^{-4}\;M$, respectively. PAL showed an optimal pH at 8.5, corresponding to the average pH range of the small intestine. It showed no loss of activity at $-80^{\circ}C$ for 5 months and possessed 93.4% of its activity under $4^{\circ}C$ for 4 wks. PAL was susceptible to chymotrypsin digestion and, to a lesser extent, to trypsin, elastase, carboxypeptidase A, and B. The trypsin and chymotrypsin cleaving sites were mutated to investigate protection from pancreatic digestion and the specific activities of these mutants were evaluated. The six mutants displayed low specific activities compared to the wild-type, suggesting that the primary trypsin and chymotrypsin cleaving sites may be essential for catalytic reaction. The PAL mutants could therefore be applied as a pretreatment modality without susceptibility to proteolytic attack, however, additional modification for enhancing enzymatic activity is needed to reduce the Phe levels effectively.
The inhibition/influences of adenine compounds on the heart have been described repeatedly by many investigators, since the first report by Druny and Szent-Gyorgyi [1929]. These studies have shown that adenosine and adenine nucleotides have an over-all effect similar to that of acetylcholine [ACh] by slowing and weakening the heartbeat. The basic cellular and membrane events underlying the inhibitory action of adenosine on sinus rate, however, are not well understood. Furthermore, the physiological role of adenosine in regulation of the heartbeat remains still to be elucidated. Therefore, this study was undertaken in order to examine the response of rabbit SA node to adenosine and to compare the response to that of ACh. Isolated SA node preparation, whole atrial pair, or left atrlal strip was used in each experiment. Action potentials of SA node were recorded through the intracellular glass microelectrodes, which were filled with 3M KCI and had resistance of 30-50 M. All experiments were performed in a bicarbonate-buffered Tyrode solution which was aerated with 3% $CO_2-97%$$O_2$ gas mixture and kept at $35^{\circ}C$. Spontaneous firing rate of SA node at 35C [Mean + SEM, n=16] was 154 + 3.3 beats/min. The parameters of action potentials were: maximum astolic potential [MDP], -731.7mV: overshoot [OS], 9 + 1.4mV; slope of pacemaker potential [SPP], 94 3.0mV/sec.Adenosine suppressed the firing rate of SA node in a dose dependent manner. This inhibitory effect appeared at the concentration of $10^{-6}M$ and was potentiated in parallel with the increase in adenosine concentration. Changes in action potential by adenosine were dose-dependent increase of MDP and decrease of SPP until $10^{-4}$. Above this concentration, however, the amplitude of action potential decreased markedly due to the simultaneous decrease of both MDP and OS. All these effects of adenosine were not affected by pretreatment of atropine [2mg/l] and propranolol [$5{\times}10^{-6}M$]. ACh [$10^{-6}M$] responses on action potential were similar to those of adenosine by increasing MDP and decreasing SPP. These effects of ACh disappeared by pretreatment of atropine [2mg/1]. Inhibition/effects of adenosine and ACh on sinus rate were enhanced synergistically with the simultaneous administration of adenosine and ACh. Marked decrease of overshoot potential was the most prominent feature on action potential. Dipyridamole [DPM], which is known to block the adenosine transport across cell membrane, definitely potentiated the action of adenosine . Adenosine suppressed the sinus rate and atrial contractility in the same dosage range, even in the reserpinized preparation. Above` results suggest that adenosine suppresses pacemaker activity, like ACh, by acting directly on the membrane of SA node, increasing MDP and decreasing SPP.
In order to control postharvest spoilage of satsuma mandarin fruits, rhizobacteria were isolated from soil samples. The Pantoea agglomerans strain 59-4 (Pa 59-4) which suppresses the decay of mandarin fruit by green and blue mold, was tested for the control efficacy and its mode of action was investigated. Pa 59-4 inhibited infection by green and blue mold on wounded mandarins, which were artificially inoculated with a spore suspension of Penicillium digitatum and P. italicum with control efficacies of 85-90% and 75-80%, respectively. The biocontrol efficacy was increased by raising the concentration of cells to between $10^8$ and $10^9\;cfu/ml$, and pretreatment with the antagonist prevented subsequent infection by green mold. The population of Pa 59-4 was increased more than 10 fold during the 24 hr incubation at $20^{\circ}C$, indicating that colonization of the wound site might prevent the infection by green mold. Despite poor antifungal activity, the Pa 59-4 isolate completely inhibited the germination and growth of P. digitatum spores at $1{\times}10^8\;cfu/ml$. We argue that the control efficacy was mediated by nutrient competition. Overall, the effective rhizobacterium, Pa 59-4, was shown to be a promising biocontrol agent for the postharvest spoilage of mandarin fruits by green and blue mold.
In order to investigate the effects of low temperature pretreatment of floral bud and plant growth regulators on anther-derived callus and shoot differentiation, anthers were cultured on 1/2 MS medium supplemented with 2,4-D, NAA, BA and TDZ. This plant depends on the plant growth regulators, for these anthers couldn't respond on 1/2 MS medium without plant growth regulators. 2,4-D was a prerequisite substance in this experiment, especially 52.6% of callus formation on MS medium with 2.0mg/L 2,4-D alone. However, the optimum medium was on 1/2 MS medium with 0.1 mg/L 2,4-D and 1.0mg/L BA for continuous growth and shoot differentiation from the anther. Calli derived from on MS medium with 2.0mg/L 2,4-D transferred to the 1/2MS medium with TDZ and BA. TDZ were less superior to BA, only one anther could produce shoot on MS media with 1.0mg/L TDZ. On the other hand, when the calli transferred to the medium with 3.0mg/L BA, adventitious shoots were proliferated, subsequently, regenerated shoots elongated from the embryogenic calli. After floral buds of one week before anthesis were incubated at $5^{\circ}C$ refrigerator for eight or fifteen days, anthers seperated from floral buds were cultured on 1/2MS medium supplemented with 0.1mg/L 2,4-D and 1.0mg/L BA. Callusing and shoot differentiation on anthers from treated at $5^{\circ}C$ for eight days were more effective than those of fifteen days or control.
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