• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.1
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• pp.143-151
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• 2010

In Vivo and In vitro antispasmodic effects of Jun-Si-Baek-Chul-San, a Traditional Korean Polyherbal Medicineconsisted of 7 types of herbs were observed in the present study. To clarify the effects of Jun-Si-Baek-Chul-San, on accelerating small intestinal movement induced by the stimulation of cholinergic neurotransmission, we evaluated the effects of Jun-Si-Baek-Chul-San on In vivo carbachol (an acetylcholinergic agent)-accelerated mice small intestinal transit and on In vitro contractions induced by low-frequency electrostimulation, KCl, histamine or acetylcholine using isolated guinea pig ileum. To induce the acceleration of mice small intestinal transit, Carbachol 1 mg/kg was once subcutaneously dosed 15min before last administration of the test drugs. In the present study, Jun-Si-Baek-Chul-San 500, 250 and 125 mg/kg or domperidone 20 mg/kg were orally pretreated on the carbachol-accelerated mice small intestinal transit once a day for 7 days and the small intestinal transit rateof activated charcoal powder were monitored. In vitro assays, Jun-Si-Baek-Chul-San1, 0.1, 0.01 and 0.001 mg/ml or domperidone $2{\times}10^{-5}M$ were treated 10min before ileal contraction was induced by filed stimulation, acetylcholine, KCl and histamine, and the % changes of contractions were observed compared to the treatment of inducer alone. In spontaneous contraction, the % changes of contractions were observed compared to treatment of vehicle alone at 10min after Jun-Si-Baek-Chul-San or domperidone treatment. The efficacy of Jun-Si-Baek-Chul-San was compared to those of domperidone. High concentration, 1 mg/ml of Jun-Si-Baek-Chul-San was found to decrease the spontaneous contraction of the isolated guinea-pig ileum. In addition, Jun-Si-Baek-Chul-San decrease contractions induced by electrostimulation, acetylcholine, histamine and KCl in the isolated guinea-pig ileum. In addition, Jun-Si-Baek-Chul-San effectively inhibited the accelerated small intestinal movement induced by carbachol stimulation of cholinergic neurotransmission in In vivo. Based on the results, although the exact molecular or action mechanism and which herbs or compound in Jun-Si-Baek-Chul-San are responsible for actions, it was concluded that Jun-Si-Baek-Chul-San normalization in the accelerated intestinal motility might be interfere with a variety of muscarinic, adrenergic and histaminic receptor activities or with the mobilization of calcium ions required for smooth muscle contraction non-specificly. Therefore, it is expected that Jun-Si-Baek-Chul-San will be promising as a prescription of clinical treatment of digestive tract disorders such as accelerated the motility of intestine, diarrhea or intestinal painful contractions.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1742-1750
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• 2015

As a novel approach for disease control and prevention, nutritional modulation of the intestinal health has been proved. However, It is still unknown whether branched-chain amino acid (BCAA) is needed to maintain intestinal immune-related function. The objective of this study was to determine whether BCAA supplementation in protein restricted diet affects growth performance, intestinal barrier function and modulates post-weaning gut disorders. One hundred and eight weaned piglets ($7.96{\pm}0.26kg$) were randomly fed one of the three diets including a control diet (21% crude protein [CP], CON), a protein restricted diet (17% CP, PR) and a BCAA diet (BCAA supplementation in the PR diet) for 14 d. The growth performance, plasma amino acid concentrations, small intestinal morphology and intestinal immunoglobulins were tested. First, average daily gain (ADG) (p<0.05) and average daily feed intake (ADFI) (p<0.05) of weaned pigs in PR group were lower, while gain:feed ratio was lower than the CON group (p<0.05). Compared with PR group, BCAA group improved ADG (p<0.05), ADFI (p<0.05) and feed:gain ratio (p<0.05) of piglets. The growth performance data between CON and BCAA groups was not different (p>0.05). The PR and BCAA treatments had a higher (p<0.05) plasma concentration of methionine and threonine than the CON treatment. The level of some essential and functional amino acids (such as arginine, phenylalanine, histidine, glutamine etc.) in plasma of the PR group was lower (p<0.05) than that of the CON group. Compared with CON group, BCAA supplementation significantly increased BCAA concentrations (p<0.01) and decreased urea concentration (p<0.01) in pig plasma indicating that the efficiency of dietary nitrogen utilization was increased. Compared with CON group, the small intestine of piglets fed PR diet showed villous atrophy, increasing of intra-epithelial lymphocytes (IELs) number (p<0.05) and declining of the immunoglobulin concentration, including jejunal immunoglobulin A (IgA) (p = 0.04), secreted IgA (sIgA) (p = 0.03) and immunoglobulin M (p = 0.08), and ileal IgA (p = 0.01) and immunoglobulin G (p = 0.08). The BCAA supplementation increased villous height in the duodenum (p<0.01), reversed the trend of an increasing IELs number. Notably, BCAA supplementation increased levels of jejunal and ileal immunoglobulin mentioned above. In conclusion, BCAA supplementation to protein restricted diet improved intestinal immune defense function by protecting villous morphology and by increasing levels of intestinal immunoglobulins in weaned piglets. Our finding has the important implication that BCAA may be used to reduce the negative effects of a protein restricted diet on growth performance and intestinal immunity in weaned piglets.

• Journal of Life Science
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• v.22 no.10
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• pp.1307-1315
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• 2012

We evaluated the effect of dietary supplements with benzoic acid on intestinal beneficial bacteria concentration and immune response of weaning pigs. Supplementation with benzoic acid at 0.5% or control diet for 35 days resulted in a higher Lactobacillus casei concentration in the cecum. Supplementation with benzoic acid at 0.5% increased concentration of L. plantarum in the cecum. Pigs with the control diet and 0.5% benzoic acid had significantly increased concentration of B. subtillis in the cecum compared to the antibiotic group, while the concentration of B. subtillis in the rectum increased in pigs given 0.3 and 0.5% benzoic acid (p<0.05). Compared with the control group, the level of interleukin-$1{\beta}$ mRNA showed a significant decrease in the proximal small intestine in pigs fed diets supplemented with benzoic acid at 0.5% or antibiotic. Feeding 0.5% benzoic acid resulted in a marked reduction in the expression of IL-6 mRNA in the middle small intestine (p<0.05). Supplementation with benzoic acid at 0.5% or antibiotic resulted in a lower level of tumor necrosis factor-mRNA in the middle intestine. Up to 0.5% benzoic acid may be included in weaning diets for improvement of intestinal beneficial bacteria, thus modulating genes of pro-inflammatory cytokines in the gastrointestinal tract.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1473-1477
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• 2007

Four-and-a-half LIM-only protein 3 (FHL3) is a member of the LIM protein superfamily and can participate in mediating protein-protein interaction by binding one another through their LIM domains. In this study, the 5'- and 3'- cDNA ends were characterized by RACE (Rapid Amplification of the cDNA Ends) methodology in combination with in silico cloning based on the partial cDNA sequence obtained. Bioinformatics analysis showed FHL3 protein contained four LIM domains and four LIM zinc-binding domains. In silico mapping assigned this gene to the gene cluster MTF1-INPP5B-SF3A3-FHL3-CGI-94 on pig chromosome 6 where several QTL affecting intramuscular fat and eye muscle area had previously been identified. Transcription of the FHL3 gene was detected in spleen, liver, kidney, small intestine, skeletal muscle, fat and stomach, with the greatest expression in skeletal muscle. The A/G polymorphism in exon II was significantly associated with birth weight, average daily gain before weaning, drip loss rate, water holding capacity and intramuscular fat in a Landrace-derived pig population. Together, the present study provided the useful information for further studies to determine the roles of FHL3 gene in the regulation of skeletal muscle cell growth and differentiation in pigs.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.12
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• pp.1729-1735
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• 2004

A total of 140 weaning pigs were used to determine the effects of digestive enzyme supplementation to corn-soybean meal diets on growth performance, physiological changes of small intestine, microorganisms and pH in the gastrointestinal tract. Two kinds of enzyme complex (A, B) were used in this experiment. Pigs were allotted in a completely random design (CRD) to five replicates with four pigs per pen. Diets and water were provided for ad libitum consumption. Treatments included 1) Control: without enzyme supplementation, 2) Enzyme A 0.05%, 3) Enzyme A 0.10%, 4) Enzyme A 0.15%, 5) Enzyme B 0.05%, 6) Enzyme B 0.10%, 7) Enzyme B 0.15% in the diets. A total of 24 crossbred barrows 25.78${\pm}$0.55 kg BW fitted with simple ileal T-cannulas were used to evaluate the effect the enzyme addition on the nutrient digstibility. Pigs were allotted 4 treatments (No enzyme, enzyme A 0.05%, enzyme A 0.1%, enzyme A 0.15%), 6 replicates according to a completely random design (CRD). Another digestibility trial was followed for enzyme complex B. Twenty pigs, average 31.92${\pm}$0.37 kg BW, fitted with simple ileal T-cannulas for digestibility trial. Neither enzyme A nor enzyme B affected on fecal or ileal digestibility of dry matter, gross energy, crude protein, crude fat and crude ash (p>0.05). The apparent fecal digestibilities of all the nutrients were higher in total feces collection method than in indirect method. At the end of feeding trial, 21 pigs were slaughtered for examining the morphological changes of small intestine and the concentration of microorganisms in the ileum and the colon. Growth performance, intestinal morphology and pH of ileum and colon were not affected by the either enzyme complex supplementation (p>0.05). These results suggested that enzyme complex A and enzyme complex B were of no benefit to early-weaned pigs when corn-soybean meal based diet was provided.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.2
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• pp.272-278
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• 2010

In this study, we cloned, sequenced and characterized porcine y+L Amino Acid Transporter-1 (y+LAT1). By screening a translated EST database with the protein sequence of the human $y^{+}$LAT1 and by using rapid amplification of cDNA ends (RACE), the full-length cDNA encoding porcine $y^{+}$LAT1 was isolated from porcine intestine RNA. It was 2,111 bp long, encoding a 511 amino acid trans-membrane glycoprotein composed of 12 transmembrane domains. The predicted amino acid sequence was found to be 91%, 90%, 87% and 87% identical to those of cattle, human, mouse and rat $y^{+}$LAT1 respectively. Real-time RT-PCR results indicated that the small intestine had the highest $y^{+}$LAT1 mRNA abundance and the lung had the lowest $y^{+}$LAT1 mRNA abundance. Baby hamster kidney (BHK) cells transfected with green fluorescent protein (GFP) tagged porcine $y^{+}$LAT1 cDNA indicated that the cellular localization of the gene product in BHK was on the plasma membrane.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1575-1581
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• 2004

A total of 96 growing barrows (Duroc${\times}$Landrace${\times}$Yorkshire) at an average BW of 20.2 kg were used to investigate the effects of montmorillonite (MMT) or copper-bearing montmorillonite (Cu-MMT) on growth performance, intestinal microflora, digestive enzyme activities of pancreas and small intestinal contents, and the apparent nutrient digestion. The pigs were allocated to three groups with 32 pigs per treatment for 42 days and the average BW at the end of the experiment was 49.7 kg. The three dietary treatments were basal diet only (control group), basal diet +1.5 g/kg MMT, and basal diet +1.5 g/kg Cu-MMT. The results showed that supplementation with Cu-MMT significantly improved growth performance as compared to control and pigs fed with Cu-MMT had higher average daily gain than those fed with MMT. As compared to control, supplementation with Cu-MMT significantly reduced the total viable counts of Escherichia coli and Clostridium in the small intestine and proximal colon. Supplementation with MMT had no significant influence on intestinal microflora, although there was a tendency for Escherichia coli and Clostridium to be lower than the control. Pigs fed with Cu-MMT had lower viable counts of Escherichia coli in colonic contents than those fed with MMT. Although supplementation with MMT improved the activities of the digestive enzymes in the small intestinal contents, the tendency was not significant. Supplementation with Cu-MMT significantly improved the activities of total protease, amylase and lipase in the small intestinal contents. Supplementation with MMT or Cu-MMT improved the apparent nutrient digestion.

• Korean Journal of Veterinary Pathology
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• v.6 no.1
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• pp.19-26
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• 2002

The purpose of this study was to investigate to potective effects against porcine epidemic diarrhea virus (PEDV) infection in piglets by administration of the PEDV antiserum orally at 2 hrs, 24hrs and 36hrs after birth. six piglets administered the antiserum were experimentally infected with PEDV at five-day-old. Control group were four piglets infected with PEDV only. Serum antibody titers against PEDV were examined by serum neutralization (SN) test, dectection for PEDV or PEDV antigen from feces and small intestines was tested by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the PEDV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of PEDV in feces and small intestines by RT-PCR were 26.2% and 16.7% in PEDV antiserum treated group and 48.1 % and 75.0% in control group, respectively. 3. The detection rate of PEDV antigen in the small intestine by IFA were 0% in PEDV antiserum treated group and 50.0% in control group, respectively. It was concluded that oral administration of antiserum against PEDV to piglets was effective in preventing PEDV infection.

• Korean Journal of Veterinary Pathology
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• v.6 no.1
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• pp.11-18
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• 2002

The purpose of this study was to investigate to potective effects against transmissible gastyoenteritis virus (TGEV) infection in piglets by administration of the TGEV antiserum orally at 5 hrs, 24hrs and 36hrs after birth. five piglets administered the antiserum were experimentally infected with TGEV at four-day-old. Control group were four piglets infected with TGEV only. Serum antibody titers against TGEV were examined by serum neutralization(SN) test, dectection for TGEV or TGEV antigen from feces and small intestines was tested by reverse transcrption-polymerase chain reaction (RT-PCR) and indirect immunoflurescence (IFA). The results obtained were as follows; 1. The piglets administered the TGEV antiserum showed higher antibody titers than those of control group and sustained during the experimental period. 2. The detection rate of TGEV in feces and small intestines by RT- PCR were 24.5% and 20.0% in TGEV antiserum treated group and 44.0% and 75.0% in control group, respectively. 3 The detection rate of TGEV antigen in the small intestine by IFA were 26.7% in TGEV antiserum treated group and 75.0% in control group, respectively. It was concluded that oral administration of antiserum against TGEV to piglets was effective in preventing TGEV infection.

• Journal of Veterinary Science
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• v.24 no.4
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• pp.53.1-53.12
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• 2023

Background: Mammalian orthoreovirus type 3 (MRV3), which is responsible for gastroenteritis in many mammalian species including pigs, has been isolated from piglets with severe diarrhea. However, the use of pig-derived cells as an infection model for swine-MRV3 has rarely been studied. Objectives: This study aims to establish porcine intestinal organoids (PIOs) and examine their susceptibility as an in vitro model for intestinal MRV3 infection. Methods: PIOs were isolated and established from the jejunum of a miniature pig. Established PIOs were characterized using polymerase chain reaction (PCR) and immunofluorescence assays (IFAs) to confirm the expression of small intestine-specific genes and proteins, such as Lgr5, LYZI, Mucin-2, ChgA, and Villin. The monolayered PIOs and three-dimensional (3D) PIOs, obtained through their distribution to expose the apical surface, were infected with MRV3 for 2 h, washed with Dulbecco's phosphate-buffered saline, and observed. Viral infection was confirmed using PCR and IFA. We performed quantitative real-time reverse transcription-PCR to assess changes in viral copy numbers and gene expressions linked to intestinal epithelial genes and antiviral activity. Results: The established PIOs have molecular characteristics of intestinal organoids. Infected PIOs showed delayed proliferation with disruption of structures. In addition, infection with MRV3 altered the gene expression linked to intestinal epithelial cells and antiviral activity, and these effects were observed in both 2D and 3D models. Furthermore, viral copy numbers in the supernatant of both models increased in a time-dependent manner. Conclusions: We suggest that PIOs can be an in vitro model to study the infection mechanism of MRV3 in detail, facilitating pharmaceutical development.