• The Korean Journal of Physiology and Pharmacology
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• v.1 no.1
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• pp.27-34
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• 1997

In the present study, it was aimed to further indentify the intracellular action mechansm of cromakalim and levcromakalim in the porcine coronary artery. In intact porcine coronary arterial strips loaded with fura-2/AM, acetylcholine caused an increase in intracellular free $Ca^{2+}$ $([Ca^{2+}]_i)$ in association with a contraction in a concentration-dependent manner. Cromakalim (1 ${\mu}M$) caused a reduction in acetylcholine-induced increased $[Ca^{2+}]_i$ not only in the mormal physiological salt solution (PSS) but also in $Ca^{2+}$-free PSS (containing 1 mM EGTA). In the skinned strips prepared by exposure of tissue to 20 .${\mu}M$ B-escin, inositol 1,4,5-trisphosphate ($IP_3$) evoked an increase in $[Ca^{2+}]_i$, but it was without effect on the intact strips. The $IP_3$-induced increase in $[Ca^{2+}]_i$ was inhibited by cromakalim by 78% and levcromakalim by 59% (1 .${\mu}M$, each). Pretreatment with glibenclamide (a blocker of ATP-sensitive $K^+$ channels, 10 .${\mu}M$) and apamin (a blocker of small conductance $Ca^{2+}$-activated $K^+$ channels, 1 .${\mu}M$) strongly blocked the effect of cromakalim and levcromakalim. However, charybdotoxin (a blocker of large conductance $Ca^{2+}$-activated $K^+$ channels, 1 .${\mu}M$) was without effect. In addition, cromakalim inhibited the $GTP{\gamma}S$ (100 .${\mu}M$, non-hydrolysable analogue of GTP)-induced increase in $[Ca^{2+}]_i$. Based on these results, it is suggested that cromakalim and levcromakalim exert a potent vasorelaxation, in part, by acting on the $K^+$ channels of the intracellular sites (e.g., sarcoplasmic reticulum membrane), thereby, resulting in decrease in release of $Ca^{2+}$ from the intracellular storage site.

• Korean Journal of Environment and Ecology
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• v.30 no.1
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• pp.130-134
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• 2016

How does $CO_2$ affect on the stomatal mechanism? The mechanism of stomatal opening by $CO_2$ is not clear as it is difficult to see $CO_2$ effect on light-induced stomatal opening. Furthermore, stomata may react differently according to the concentration of $CO_2$. The significance of the possible endogenous rhythms must consider to understand on $CO_2$-related response. It is clear that $CO_2$ has an effect on the accumulation of osmotic materials which determines the degree of stomatal apertures because it is known that stomata open in the condition of the reduced $CO_2$ concentration. However, it is not fully understood how $CO_2$ leads to the stomatal opening. It has been thought that $CO_2$ can not affect on the ion fluxes which determines the increase of osmotic potential in guard cells. However, in this study, the changes of guard cell membrane permeability by $CO_2$ have been focused on. There are many reports that $CO_2$ related reactions are dominant when the leaf is exposed to certain a mount of $CO_2$. The hypothesis of the stomatal opening by light is based on the increase of osmotic materials in guard cells including $K^+$, $Cl^-$, sucrose and $malate^{2-}$. It was reported that $CO_2$ induced a big hyperpolarization indicating that $H^+$ was extruded to the cell outside. It was also found that $CO_2$ caused guard cell membrane hyperpolarization in the intact leaf up to 3 or 4 times higher than that of light induced membrane hyperpolarization. These results represent that $CO_2$ can affect on the change of physical characteristics which affects on the change of the membrane permeability.

• Molecules and Cells
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• v.24 no.3
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• pp.378-387
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• 2007

The Bcl-2 family of proteins interacts at the mitochondria to regulate apoptosis. However, the anti-apoptotic Bcl-2 and $Bcl-X_L$ are not completely localized to the mitochondria. In an attempt to generate Bcl-2 and $Bcl-X_L$ chimeras that are constitutively localized to the mitochondria, we substituted their C-terminal transmembrane tail or both the C-terminal transmembrane tail and the adjacent loop with the equivalent regions from Bak or Bax mutant (BaxS184V) as these regions determine the mitochondrial localization of Bak and Bax. The effects of these substitutions on subcellular localization and their activities were assessed following expression in HeLa and CHO K1 cells. The substitution of the C-terminal tail or the C-terminal tail and the adjacent loop of Bcl-2 with the equivalent regions from Bak or the Bax mutant resulted in its association with the mitochondria. This change in subcellular localization of Bcl-2 chimeras triggered cells to undergo apoptotic-like cell death. The localization of this Bcl-2 chimera to the mitochondria may be associated with the disruption of mitochondrial membrane potential. Unlike Bcl-2, the loop structure adjacent to the C-terminal tail in $Bcl-X_L$ is crucial for its localization. To localize the $Bcl-X_L$ chimeras to the mitochondria, the loop structure next to the C-terminal tail in $Bcl-X_L$ protein must remain intact and cannot be substituted by the loop from Bax or Bak. The chimeric $Bcl-X_L$ with both its C-terminal tail and the loop structure replaced by the equivalent regions of Bak or Bax mutant localized throughout the entire cytosol. The $Bcl-X_L$ chimeras that are targeted to the mitochondria and the wild type $Bcl-X_L$ provided same protection against cell death under several death inducing conditions.

• Journal of Environmental Science International
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• v.6 no.2
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• pp.125-131
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• 1997

The mechanism of stomatal closing in response to $O_2$ was indirectly investigated by using $H_2O_2$ which is the intermediate product of $O_2$ metabolites. Stomata in epidermal strips close in response to $H_2O_2$. The effect of $H_2O_2$ on stomatal closing was dependent on the concentration of $H_2O_2$. 10 ppm $H_2O_2$ showed a clear effect on stomatal closing and 1000 ppm $H_2O_2$ induced complete stomatal closing after the treatment of 3 hours. Stomatal closing by $H_2O_2$ in intact leaf was also observed by measuring the diffusion resistance with porometer. It was found that the stomatal closing by $H_2O_2$ was not mediated by $Ca^{2+}$, and that was a different result observed in stomatal closing by water stress. Reversely, $Ca^{2+}$ showed a great inhibition on stomatal closing. The leakage of K+ in epidermal strips was doubled in response to $H_2O_2$ when it was campared to the control. 10 ppm $H_2O_2$ decreased photosynthetic activity. Fv/Fm representing the activity of Photosystem II was reduced about 4 % in 10 ppm $H_2O_2$ and 8 % in 100 ppm $H_2O_2$ In the treatment of 1.5 hour. However, stomatal closing by 10 ppm $H_2O_2$ was reduced about 56 %. According1y, it can be suggested that stomatal closing by $H_2O_2$ is related with the decrease of photosynthetic activity, but it was chiefly induced by the change of the membrane permeability. Key words Commelina communis, stomatal closing, $H_2O_2$, $Ca^{2+}$, photosynthesis.

• Archives of Plastic Surgery
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• v.34 no.6
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• pp.765-770
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• 2007

Purpose: Peritendinous adhesion is one of the most notorious complication after the flexor tendon injury. In this study, $Alloderm^{(R)}$(LifeCell Corp., Branchburg, N.J.), which is the decellularized human dermal analogue with its intact native basement membrane components, was used for the prevention of peritendinous adhesions following flexor tendon repair. Methods: Thirty New Zealand white male rabbits were divided equally into 3 groups. In all groups, the flexor digitorum profundus of the third finger of the right back foot was cut totally and repaired by modified Kessler suture technique. Following tendon repair, $Alloderm^{(R)}$ was wrapt around the repaired tendon in the first group and sodium hyaluronate gel was sprayed to the operation field in the second group. In the control group, no external material was applied. The right back foot were immobilized for 6 weeks to optimize the formation of adhesion ingrowth. After death, the third finger that repaired tendons and sheaths was removed en bloc. We checked range of motion. and studied histologically for all groups. Results: The experimental groups had better range of motion than the control group. We checked that the range of motion was 73.5 degrees in $Alloderm^{(R)}$ group, 55.9 degrees in the hyaluronic acid group, and 38.3 degrees in the control group. in the histological study, the experimental group had less adhesions compared with the control group. Conclusion: This study concludes that $Alloderm^{(R)}$ can decrease peritendinous adhesions following flexor tendon repairs in rabbits. We think the method could be used in clinical cases.

• Journal of Veterinary Clinics
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• v.31 no.2
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• pp.112-116
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• 2014

A 2-month-old intact male Pitbull terrier (weighing 1.01 kg) were referred with primary complaints of severe abdominal distension, exercise intolerance, retarded growth, yellow nasal discharge and anorexia. Diagnostic imaging studies found enlarged right atrium partitioned by abnormal membrane, severe pulmonic systolic jets (5.66 m/s of peak velocity) and right-to-left blood shunt at the inter-atrial septum. Based on clinical and diagnostic findings, the case was diagnosed as cor triatriatum dexter complicated with severe pulmonic stenosis and right-to-left shunted patent foramen ovale. Either surgical or interventional therapy has not been attempted, because of unstable patient's condition. Using blood transfusion, oxygen supply and cardiac medications (i.e., sildenafil, spironolactone, enalapril), the dog was recovered.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.179-190
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• 1998

Prolactin (PRL) surge in cycling rats at proestrous afternoon has previously been reported as an inducer of apoptotic cell death of luteal cells. This death-inducing action of PRL seeins unusual, because PRL can he categorized as a cell-survival factor, if other known physiological functions of PRL are taken into account. In this study, the apoptotic action of PRL was assessed in cultured cells prepared from rat luteal tissue and underlying molecular /cellular mechanism of PRL-induced luteolysis was analyzed. The latest crop of corpora lutea (CLs) were enucleated from rat ovaries at 18:00 h on the proestrous day before the next ovulation. Donor rats were pretreated with CB154, a dopamine agonist, in order to he exempted from the endogenous PRL surge. The harvested GLs were dispersed and cultured with or without PRL (2$\mu$g /ml) for 24 or 48 h. An addition of PRL to the culture medium changed the parameters indicative of cell death via apoptosis: a decrease in cell viability (MTT) and an increase in chromatin condensation. Most of the DNA breakdown in nuclei induced by PRL occurred in steroidogenic cells which were identified by 3$\beta$-HSD activity staining, and the number of 3$\beta$-HSD-positivecells were significantly decreased. Interestingly, most of the cells with an apoptotic nucleus adhered to one or more intact and seemingly non-steroidogenic cells. Because the expression of Fas has heen shown to be abundant in murine ovary, and Fas is known to have an exact physiological role in occurrence of apoptotic cell death, the membrane form-Fas ligand (rnFasL) was quantified in the cell lysate. An addition of PRL increased expression of mFasL. Moreover, an addition of concanavalin A (ConA), a T-cell specific activator, in place of PRL, enhanced the apoptotic parameters. Cumulatively, the apoptotic PRL action was addressed to cells unknown than steroidogenic lute~ cells. The most prohable candidate for the direct target cells is Tcells in the luteal tissue that can express mFasL in response to PRL.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.175-175
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• 1996

The spinal dorsal horn is the area where primary afferent fibers terminate and cutaneous sensory information is Processed. A number of putative neurotransmitter substances, including excitatory and inhibitory amino acids and peptides, are present in this region and sites and cellular mechanisms of their actions have been a target of numerous studies. In this study, single neurons were acutely isolated and the properties of whole cell current and responses to excitatory and inhibitory neurotransmitters were studied by the patch clamp method. Young rats (7-14 days) were anesthetized with diethyl-ether, and the lumbar spinal cord was excised and cut transversely at a thickness of 30$\mu\textrm{m}$ by Vibroslicer. The treatment of spinal slices with low concentration of proteases (pronase and thermolysin 0.75 mg/$m\ell$) and mechanical dissociation yielded isolated neurons with near intact morphology. Multipolar, ellipsoidal and bipolar, and pyramidal cells were shown. By applying step voltage pulses to neurons held at -70 mV, two types of inward currents and one outward currents observed. The fast activating and inactivating inward current was the Na$\^$+/ current because of its fast kinetics and blocking by 0.5${\mu}$M TTX, a specific blocker of Na$\^$+/ channel. The second type of inward currents were sustained. Based on their kinetics and current-voltage relations, it was likely that the second type of inward current was the voltage-dependent Ca$\^$2+/ current. In the presence of TTX, the steady-state currents mainly represented outward K$\^$+/ current which looked like the delayed rectifier K$\^$+/ current. In addition, the membrane currents produced by agonist of excitatory amino acid (EAA) receptor and the endogenous transmitter candidate L-glutamate were recorded in isolated whole-cell voltage clamped neurons as well as responses to inhibitory amino acids (${\gamma}$-amino butyric acid, glycine). Drugs were applied by a method that allows complete exchange of the solution within 1 sec; an infinite number of solutions can be applied to a single cell.

• Journal of Microbiology and Biotechnology
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• v.1 no.1
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• pp.22-30
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• 1991

In order to secrete the Bacillus subtilis cytidine deaminase (CDase, cytidine/2'-deoxycytidine deaminase) encoded by the B. subtilis cdd gene in E. coli by the aid of signal sequences, the cdd gene was fused in-frame to either amyE or penP signal sequences and the gene expression and CDase localization were examined. For the penP signal sequence::cdd fusion, the cdd gene with 9 amino acids truncated from the 5'-terminus was fused in-frame to the signal sequence, then the $cdd^{+}$ colonies were not occurred from the minimal plate by cdd complementation. The result suggests that 9 amino acids on the $NH_2-terminal$ of CDase have an essential function in the enzyme activity. The hybrid protein obtained by fused gene amyE signal sequence::cdd structural gene gave $cdd^{+}$ phenotype and about half of the total CDase activity was found to be secreted in the periplasm of E. coli transformant JF611/pSO202. The periplasmic CDase activity of JF611 harboring pSO52 containing the intact cdd gene was considerablely lower than that of the cells harboring pSO202 carrying the hybrid cdd gene. This suggests that the CDase was secreted to the periplasm through the cytoplasmic membrane by the aid of the amyE signal sequence in the E. coli transformant.

• Journal of Embryo Transfer
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• v.31 no.4
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.