• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.743-748
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• 2014

The objective of this study was to determine the molecular characteristics of the horse vascular endothelial growth factor alpha gene ($VEGF{\alpha}$) by constructing a phylogenetic tree, and to investigate gene expression profiles in tissues and blood leukocytes after exercise for development of suitable biomarkers. Using published amino acid sequences of other vertebrate species (human, chimpanzee, mouse, rat, cow, pig, chicken and dog), we constructed a phylogenetic tree which showed that equine $VEGF{\alpha}$ belonged to the same clade of the pig $VEGF{\alpha}$. Analysis for synonymous (Ks) and non-synonymous substitution ratios (Ka) revealed that the horse $VEGF{\alpha}$ underwent positive selection. RNA was extracted from blood samples before and after exercise and different tissue samples of three horses. Expression analyses using reverse transcription-polymerase chain reaction (RT-PCR) and quantitative-polymerase chain reaction (qPCR) showed ubiquitous expression of $VEGF{\alpha}$ mRNA in skeletal muscle, kidney, thyroid, lung, appendix, colon, spinal cord, and heart tissues. Analysis of differential expression of $VEGF{\alpha}$ gene in blood leukocytes after exercise indicated a unimodal pattern. These results will be useful in developing biomarkers that can predict the recovery capacity of racing horses.

• International Journal of Oral Biology
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• v.43 no.1
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• pp.43-51
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• 2018

$K^+$ channels are key components of the primary and secondary basolateral $Cl^-$ pump systems, which are important for secretion from the salivary glands. Paroxetine is a selective serotonin reuptake inhibitor (SSRI) for psychiatric disorders that can induce QT prolongation, which may lead to torsades de pointes. We studied the effects of paroxetine on a human $K^+$ channel, human ether-a-go-go-related gene (hERG), expressed in Xenopus oocytes and on action potential in guinea pig ventricular myocytes. The hERG encodes the pore-forming subunits of the rapidly-activating delayed rectifier $K^+$ channel ($I_{Kr}$) in the heart. Mutations in hERG reduce $I_{Kr}$ and cause type 2 long QT syndrome (LQT2), a disorder that predisposes individuals to life-threatening arrhythmias. Paroxetine induced concentration-dependent decreases in the current amplitude at the end of the voltage steps and hERG tail currents. The inhibition was concentration-dependent and time-dependent, but voltage-independent during each voltage pulse. In guinea pig ventricular myocytes held at $36^{\circ}C$, treatment with $0.4{\mu}M$ paroxetine for 5 min decreased the action potential duration at 90% of repolarization ($APD_{90}$) by 4.3%. Our results suggest that paroxetine is a blocker of the hERG channels, providing a molecular mechanism for the arrhythmogenic side effects of clinical administration of paroxetine.

• The Korean Journal of Physiology
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• v.22 no.1
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• pp.1-12
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• 1988

The changes in membrane potential and intracellular $Ca^{2+}$ activities along with adenosine triphosphate (ATP) and creatine phosphate (CP) contents of guinea pig hearts were studied during the cardioplegia and recovery phase. The results obtained were summarized as follows: 1) A low concentration of $Ca^{2+}$ in the cardioplegic solution caused significant changes in membrane potential during the cardioplegia and action potential configurations of the first beat after cardioplegia. 2) Intracellular $Ca^{2+}$ activities were decreased significantly during cardioplegia and increased when the cardiac tissue was reperfused with the normal Tyrode solution. The magnitude of increase in intracellular $Ca^{2+}$ activities was larger in the 0.1 mM $Ca^{2+}-cardioplegic$ solution than the 1.8 mM $Ca^{2+}-solution$. 3) ATP and CP contents of cardiac ventricular tissue were decreased during the cardioplegia, regardless of $Ca^{2+}$ concentration in the cardioplegic solutions, and CP contents were recovered with the reperfusion of normal Tyrode solution faster than those of ATP. And, there were no significant differences in the recovery of CP contents with different concentrations of $Ca^{2+}$ in the cardioplegic solutions tested, while the recovery of ATP contents was faster with $15mM\;K^{+},\;0.1mM\;Ca^{2+}$ cardioplegic solution. These results suggest that the $Ca^{2+}$ concentration in the cardioplegic solution is one of major factors influencing the recovery of cardiac tissue from the cardioplegia.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.3
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• pp.353-359
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• 1998

In gastrointestinal smooth muscle, muscarinic stimulation by carbachol (CCh) activates nonselective cation channel current ($I_{CCh}$) which is facilitated by intracellular [$Ca^{2+}$] increase. Caffeine is widely used in experiments to mobilize $Ca^{2+}$ from intracellular stores. This study shows a strong inhibitory effect of caffeine on $I_{CCh}$ in guinea-pig gastric myocyte. In this study, the underlying mechanism of the inhibitory effect of caffeine was investigated. $I_{CCh}$ was completely suppressed by the addition of caffeine (10 mM) to the superfusing solution. Inhibition of $I_{CCh}$ by caffeine was not related to the intracellular cAMP accumulation which was expected from the phosphodiesterase-inhibiting effect of caffeine. The blockade of $InsP_3-induced$ $Ca^{2+}$ release by heparin had no significant effects on the activation of $I_{CCh}$. When the same cationic current had been induced by intracellular dialysis of $GTP[{\gamma}S]$ in order to bypass the muscarinic receptor, the inhibitory effect of caffeine was significantly attenuated. The results of this study indicate that both intracellular signalling pathways for $I_{CCh}$, proximal and distal to G-protein activation, are suppressed by caffeine. A major inhibition was observed at the proximal level.

• Korean Journal of Agricultural Science
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• v.48 no.2
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• pp.353-365
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• 2021

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

• Journal of Animal Science and Technology
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• v.65 no.2
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• pp.365-376
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• 2023

Pig breeding management directly contributes to the profitability of pig farms, and pregnancy diagnosis is an important factor in breeding management. Therefore, the need to diagnose pregnancy in sows is emphasized, and various studies have been conducted in this area. We propose a computer-aided diagnosis system to assist livestock farmers to diagnose sow pregnancy through ultrasound. Methods for diagnosing pregnancy in sows through ultrasound include the Doppler method, which measures the heart rate and pulse status, and the echo method, which diagnoses by amplitude depth technique. We propose a method that uses deep learning algorithms on ultrasonography, which is part of the echo method. As deep learning-based classification algorithms, Inception-v4, Xception, and EfficientNetV2 were used and compared to find the optimal algorithm for pregnancy diagnosis in sows. Gaussian and speckle noises were added to the ultrasound images according to the characteristics of the ultrasonography, which is easily affected by noise from the surrounding environments. Both the original and noise added ultrasound images of sows were tested together to determine the suitability of the proposed method on farms. The pregnancy diagnosis performance on the original ultrasound images achieved 0.99 in accuracy in the highest case and on the ultrasound images with noises, the performance achieved 0.98 in accuracy. The diagnosis performance achieved 0.96 in accuracy even when the intensity of noise was strong, proving its robustness against noise.

• Journal of Animal Science and Technology
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• v.47 no.1
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• pp.19-28
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• 2005

The objectives of the current study were to develop polyclonal antibodies in sheep against adipocyte plasma membrane(APM) proteins isolated from swine, to investigate tissue specificity, and to determine cytotoxic effects of antiserum on swine adipocytes. Plasma membrane proteins from adipocyte, brain, heart, kidney, liver, and spleen were isolated using a 32% sucrose gradient. Adult male sheep was immunized three times at three week interval with the purified swine APM proteins. Antiserum was taken from immunized sheep at 10, 12, and 14 days after the third immunization. Antiserum expressed strong reactivity with APM proteins determined by enzyme-linked immunosorbent assay(ELISA), and the reactivity could be detected at dilutions in excess of 1 : 81,000. Antiserum showed very low binding affinity with proteins isolated from brain, heart, kidney, liver, or spleen. Tissue specificity of the antiserum was reconfirmed by Western immunoblotting using anti-sheep immunoglobulin G•alkalinephosphatase conjugate as a secondary antibody. The reactivity of antiserum to the external surface of fixed swine adipocytes was confmned by an immunohistochemical technique using anti-sheep immunoglobulin G-FITC. Confluent swine adipocytes in culture were lysed by antiserum treatment and cytosolie lactate dehydrogenase(LDH) was released as a dose-dependent patterns while adipocytes treated with normal sheep serum maintained their integrity and expressed low level of LDH. These results implicate that fat contents in the pigs can be reduced by immunological methods.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.1
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• pp.23-30
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• 2019

Objective: Recent studies have demonstrated that lin-28 homolog B (LIN28B)/miRNA let-7 (let-7) plays a role in the regulation of pubertal onset in mammals. However, the role of LIN28B/let-7 in the onset of ovine puberty remains unknown. We cloned the Duolang sheep Lin28B cDNA sequence, detected the expression change of LIN28B, let-7a and let-7g in hypothalamus, pituitary and ovary tissues at three different pubertal stages. Methods: The reverse transcriptase polymerase chain reaction (RT-PCR) was used to clone the cDNA sequence of LIN28B gene from Duolang sheep and the bioinformatics methods were applied to analyze the amino acid sequence of LIN28B protein. The mRNA expression levels of the LIN28B gene at different pubertal stages were examined by real time RT-PCR. Results: LIN28B cDNA of Duolang sheep was cloned, and two transcripts were obtained. The amino acid sequence of transcript 1 shares 99.60%, 98.78%, and 94.80% identity with those of goat, wild yak and pig, respectively. Strong LIN28B mRNA expression was detected in the hypothalamus, pituitary, ovary, oviduct and uterus, while moderate expression was found in the liver, kidney, spleen and heart, weak expression was observed in the heart. No expression was found in the lungs. Quantitative real-time PCR (QPCR) and western-blot analysis revealed that the LIN28B was highly expressed in the hypothalamus and ovary at prepuberty stages, and this expression significantly decreased from the prepuberty to puberty stages (p<0.05). Markedly increased levels of mRNA expression were detected in the pituitary from prepuberty to puberty (p<0.05) and then significantly decreased from puberty to post-puberty (p<0.05). The expression levels of let-7a and let-7g showed no significant changes among different pubertal stages (p>0.05). Conclusion: These results provided a foundation for determining the functions of LIN28B/let-7 and their role in the onset of sheep puberty.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.3
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• pp.452-458
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• 2019

Objective: The study was carried to determine the interaction effects of pen enrichment and sex on behavioral activities, skin lesions and physiology of Windsnyer pigs. Methods: Forty-eight growing Windsnyer pigs of both sex, with an average initial body weight of 21.6 (${\pm}9.01$) kg were used. Four pigs were randomly assigned to either enriched or barren pens at a stocking density of $0.35m^2/pig$. Enriched pens contained 2 L bottles filled with stones and suspended at head level on ropes stretching across the pens. In addition, two plastic balls (90 mm in diameter) and 500 mL bottles (235 mm long) were placed on the floor of each enriched pen. Results: Pigs in barren environments had higher heart rates (p<0.001) than those in enriched pens. There was an interaction of pen environment and sex on rectal temperature (p<0.001). Females in enriched pens had higher rectal temperatures (p<0.05) than females in barren pens. There was no interaction of pen environment and sex on time spent eating and drinking (p>0.05). Time spent bullying was influenced (p<0.05) by pen environment and sex. Female pigs in barren environment spent more time on bullying than females in enriched pens. There was an interaction of pen environment and sex on time spent lying down and walking (p<0.05). Female pigs in enriched pens spent more time lying down than females in barren pens. Males in barren pens spent more time walking than males in enriched pens while no effect of pen environment was observed in females. There was an interaction of pen environment and sex on the number of skin lesions in the head, neck and shoulder region and other parts of the body (p<0.05). Conclusion: It was concluded that pen enrichment reduced the number of skin lesions and anti-social behaviors, especially for female pigs. There is a need, therefore of housing indigenous pigs under confinement.

• The Korean Journal of Physiology
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• v.23 no.1
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• pp.109-117
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• 1989

The changes in adenosine triphosphate (ATP), creatine phosphate (CP) and lactic acid (LA) contents of guinea pig hearts were studied during the cardioplegia and recovery phase. 1) ATP and CP contents in cardiac ventricular tissue were decreased during the cardioplegia, regardless of $Ca^{2+}$ concentration in the cardioplegic solutions, and CP contents were recovered with the reperfusion of normal Tyrode solution faster than those of ATP. And there were no significant differences in the recovery of CP contents with different concentration of $Ca^{2+}$ in the cardioplegic solutions tested, while the recovery of ATP contents was faster with 15 mM $K^{+}$, 0.1 mM $Ca^{2+}$ cardioplegic solutions. 2) LA contents were increased during the cardioplegia and decreased with the reperfusion of normal Tyrode solution. 3) The more recovery time (up to 3 hrs), the more CP contents were recovered with the reperfusion of normal Tyrode solution faster than those of ATP. And LA contents were decreased as the duration of recovery time. These results suggest that $Ca^{2+}$ and $K^{+}$ concentration in the cardioplegic solution is one of the major factors influencing the recovery of cardiac tissue from the cardioplegia.