• Asian-Australasian Journal of Animal Sciences
• /
• v.23 no.5
• /
• pp.640-648
• /
• 2010

Insulin-responsive glucose transporter 4 (GLUT4) is a member of the glucose transporter family and mainly presents in skeletal muscle and adipose tissue. To clarify the molecular structure of porcine GLUT4, RACE was used to clone its cDNA. Several cDNA clones corresponding to different regions of GLUT4 were obtained by amplifying reverse-transcriptase products of total RNA extracted from Landrace porcine skeletal muscles. Nucleotide sequence analysis of the cDNA clones revealed that porcine GLUT4 cDNA was composed of 2,491 base pairs with a coding region of 509 amino acids. The deduced amino acid sequence was over 90% identical to human, rabbit and cattle GLUT4. The tissue distribution of GLUT4 was also examined by Real-time RT-PCR. The mRNA expression abundance of GLUT4 was heart>liver, skeletal muscle and brain>lung, kidney and intestine. The developmental expression of GLUT4 and insulin receptor (IR) was also examined by Real-time RT-PCR using total RNA extracted from longissimus dorsi (LM), semimembranosus (SM), and semitendinosus (SD) muscle of Landrace at the age of 1, 7, 30, 60 and 90 d. It was shown that there was significant difference in the mRNA expression level of GLUT4 in skeletal muscles of Landrace at different ages (p<0.05). The mRNA expression level of IR also showed significant difference at different ages (p<0.05). The developmental change in the mRNA expression abundance of GLUT4 was similar to that in IR, and both showed a higher level at birth and 30 d than at other ages. However, there was no significant tissue difference in the mRNA expression of GLUT4 or IR (p>0.05). These results showed that the nucleotide sequence of the cDNA clones was highly identical with human, rabbit and cattle GLUT4 and the developmental change of GLUT4 mRNA in skeletal muscles was similar to that of IR, suggesting that porcine GLUT4 might be an insulin-responsive glucose transporter. Moreover, the tissue distribution of GLUT4 mRNA showed that GLUT4 might be an important nutritional transporter in porcine skeletal muscles.

• Asian-Australasian Journal of Animal Sciences
• /
• v.32 no.2
• /
• pp.170-175
• /
• 2019

Objective: Uncoupling protein 3 gene (UCP3) is a candidate gene associated with the meat quality of pigs. The aim of this study was to explore the regulation mechanism of UCP3 expression and provide a theoretical basis for the research of the function of porcine UCP3 gene in meat quality. Methods: Bisulfite sequencing polymerase chain reaction (PCR) and quantitative real-time PCR (Q-PCR) were used to analyze the methylation of UCP3 5′-flanking region and UCP3 mRNA expression in the adipose tissue or skeletal muscle of three pig breeds at different ages (1, 90, 210-day-old Putian Black pig; 90-day-old Duroc; and 90-day-old Dupu). Results: Results showed that two cytosine-guanine dinucleotide (CpG) islands are present in the promoter region of porcine UCP3 gene. The second CpG island located in the core promoter region contained 9 CpG sites. The methylation level of CpG island 2 was lower in the adipose tissue and skeletal muscle of 90-day-old Putian Black pigs compared with 1-day-old and 210-day-old Putian Black pigs, and the difference also existed in the skeletal muscle among the three 90-day-old pig breeds. Furthermore, the obvious changing difference of UCP3 mRNA expression was observed in the skeletal muscle of different groups. However, the difference of methylation status and expression level of UCP3 gene was not significant in the adipose tissue. Conclusion: Our data indicate that UCP3 mRNA expression level was associated with the methylation status of UCP3 promoter in the skeletal muscle of pigs.

• BMB Reports
• /
• v.30 no.2
• /
• pp.106-112
• /
• 1997

The effect of cryopreservation on extracellular matrix was studied with the ultimate objective of permiting a prediction of the tendency of aorta conduit tissue to calcify following transplantation. Cryopreserved and fresh porcine aorta conduit tissues were extracted using guanidine-hydrochloride (Gdn-HCl) followed by sequential digestion of the tissues with collagenase, elastase, and papain. Glycosaminoglycans (GAGs) of the proteoglycans (PGs) were isolated and quantitated. Gdn-HCl extracted about 61% and 62% of the total GAG (proteoqlycan) material from cryopreserved and fresh tissues, respectively. Collagenasesolubilized proteoglycans from Gdn-HCl extracted tissue represented 20% and 13%, respectively, of the total GAGs present in cryopreserved and fresh tissues. Subsequent elastase hydrolysis of collagenase-digested tissue released about 11% of total GAGs from cryopreserved tissue and 16% from fresh tissue. The remaining 8%, from cryopreserved tissue, and 9%, from fresh tissue, of the total GAGs were obtained after using a papain hydrolysis. There was essentially no difference between fresh and cryopreserved tissues in the relative distribution of proteoglycans in the extracts and digestions except in the initial digestion step where more proteoglycans were obtained from collagenase solubilization of cryopreserved tissue than fresh tissue (p<0.05). The histologic status of the fresh and cryopreserved porcine aortic conduit did not differ markedly. The normal tissue architecture was not affected markedly by the cryopreservation procedure as neither alteration of elastic structure, fibrous proteins nor alteration of nuclear distribution or smooth muscle cell morphology was detected. Quantitative tissue mineral studies revealed that the mean calcium content of the cryopreserved aorta conduit tissue $(165{\pm}3\;{\mu}g/g\;wet\;tissue)$ was higher than that of the fresh tissue $(105{\pm}4\;{\mu}g/g\;wet\;tissue)$ $(p<0.05)$. The mean phosphorus content was $703{\pm}35\;{\mu}g$ wet tissue from cryopreserved tissue and $720{\pm}26\;{\mu}g$ wet tissue from fresh tissue. The study indicates that there is no significant alteration in the distribution of PGs in properly cryopreserved tissue, but the total calcium level appears to be increased in tissue cryopreserved by the cryopreservation process used in this study.

• Korean Journal of Veterinary Research
• /
• v.60 no.4
• /
• pp.209-213
• /
• 2020

This study examined the residue of dl-methylephedrine hydrochloride (MEP) on the muscle of pigs administered orally with MEP 12 g/ton feed for seven consecutive days. Twenty healthy cross swine were administered MEP. Four treated animals were selected arbitrarily to be sacrificed at 1, 2, 3, 4, and 5 days after treatment. MEP residue concentrations in the muscle were determined by liquid chromatography coupled with tandem mass spectrometry. The drug was extracted from muscle samples using 10 mM ammonium formate in acetonitrile followed by clean-up with n-hexane. The analyte was separated on an XBridgeTM hydrophilic interaction liquid chromatography column using 10 mM ammonium formate in deionized distilled water and acetonitrile. The correlation coefficient (R2) of the calibration curve was 0.9974, and the limits of detection and quantification were 0.05 and 0.15 ㎍/kg, respectively. The recoveries at three spiking levels were 94.5-101.2%, and the relative Standard Deviations was less than 4.06%. In the MEP-treated group, MEP residues on one day post-treatment were below the maximum residue limit in the muscle. The developed method is sensitive and reliable for the detection of MEP in porcine muscle tissues. Furthermore, it exhibits low quantification limits for animal-derived food products destined for human consumption.

• Korean Journal of Food Science and Technology
• /
• v.23 no.1
• /
• pp.6-14
• /
• 1991

Intramuscular lipid of longissimus dorsi muscle(white muscle), soleus muscle(red muscle) and cardiac muscle were autooxidized at $37^{\circ}C$ for 20 days, and the rancidity development and the effect of various factors on rancidity development were compared. Although the myoglobin content of red muscle was about 5 times as high as that of white muscle, the degree of autooxidation occurred in intramuscular lipid did not differ between red muscle and white muscle, when they had the same lipid content. Accordingly, it was suggested that the susceptibility of muscle tissues to lipid oxidation depends mainly on the lipid content of muscle tissue, regardless of muscle types. Lipid oxidation was not a major quality deterioration for dried-pork product, when it contained adequate amount of sodium nitrite and was air-tight vacuum-packed.

• Biomaterials and Biomechanics in Bioengineering
• /
• v.2 no.4
• /
• pp.225-235
• /
• 2015

Treatments for asthma are largely pharmaceutical, with some therapies also utilising alternative breathing techniques. The objective of both medical and alternative methods is to relax contracted airway smooth muscle (ASM). In normal subjects, tidal breathing- and deep inspiration-oscillations are believed to have a bronchodilatory effect. Similarly, application of length oscillations to isolated, contracted ASM also elicits muscle relaxation. As a means of investigating more-effective alternative treatment methods for contracted airways, we analyse the combined effects of bronchodilators and length oscillations on isolated, contracted ASM. The contractile state of the muscle tissue prior to treatment is of primary interest. Thereafter, the effect of applying a combination of small superimposed length oscillations with tidal breathing-like oscillations to ASM is studied alone and in combination with a common bronchodilator, isoproterenol (ISO). This work suggests that relaxation of isolated, contracted ASM following application of combined oscillations and ISO is larger than treatments of either combined oscillations or ISO alone. Further, the observed oscillation-associated relaxation is found to be amplitude- rather than frequency-dependent. This study gives additional insight into the role of oscillations and bronchodilators on contracted airways.

• Archives of Plastic Surgery
• /
• v.38 no.5
• /
• pp.559-566
• /
• 2011

Purpose: The use of tissue expander/implant in breast reconstruction using tissue expander-implant is one of the most common surgical procedures. The use of AlloDerm as a sling to reestablish the lower pole of the pectoralis major muscle results a decrease in morbidity compared with more invasive procedures. However the use of AlloDerm is more expensive than other options. We decided to compare AlloDerm with Permacol, which has been safely used in human body reconstruction and is less costly than AlloDerm. Methods: After mastectomy, the inferolateral origin of the pectoralis major muscle was elevated. Either AlloDerm or Permacol was sutured to the chest wall at the level of the previously marked inframammary fold. The lower border of the pectoralis major muscle and the upper portion of the crescent-shaped piece of either AlloDerm or Permacol was sutured together using a tension free technique, and a tissue expander was subsequently inserted into the subpectoral-subAlloDerm (or Permacol) dual pocket. Results: AlloDerm was used in twenty-one patients (28 breasts) and Permacol was used in six patients (11 breasts) for tissue expander-implant breast reconstruction. During the mean follow-up period of 17 months (8~25 months). Two infections (7%) occurred in AlloDerm cases and four infections (36%) occurred in Permacol cases. Conclusion: This study is the first comparison of tissue expander/implant breast reconstruction using AlloDerm and Permacol. The use of Permacol resulted in more postoperative infection compared with the use of AlloDerm. This report is still limited with the small number of cases studied.

• Reproductive and Developmental Biology
• /
• v.37 no.4
• /
• pp.219-224
• /
• 2013

Muscular satellite cell (SC), which is stem cell of postnatal pig, is an important for study of differentiation into adipogenesis, myogenesis, and osteoblastogenesis. In this study, we isolated and examined from pig muscle tissue to determine capacity in proliferate, differentiate, and expression of various genes. Porcine satellite cells (PSC) were isolated from semimembranosus (SM) muscles of 90~100 days old pigs according to standard conditions. The cell proliferation increased in multi-potent cell by Masson's, oil red O, and Alizarin red staining respectively. We performed the expression levels of differentiation related genes using real-time PCR. We found that the differentiation into adipocyte increased expression levels of both fatty acid binding protein 4 (FABP4) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) genes (p<0.01). Myocyte increased the expression levels of the myosin heavy chain (MHC), myogenic factor 5 (Myf5), myogenic regulatory factor (MyoD), and Myogenic factor 4 (myogenin) (p<0.01). Osteoblast increased the expression levels of alkaline phosphatase (ALP) (p<0.01). Finally, porcine satellite cells were induced to differentiate towards adipogenic, myogenic, and osteoblastogenic lineages. Our results suggest that muscle satellite cell in porcine may influence cell fate. Understanding the progression of PSC may lead to improved strategies for augmenting meat quality.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.7
• /
• pp.954-960
• /
• 2008

MiRNAs (microRNAs) are a class of small non-coding RNA molecules of ~21 nucleotides that down- regulate the expression of target genes at post-transcriptional level. In this study, we first accomplished a preliminary scan of miRNA expression using 65 and 90 day fetal pig skeletal muscle samples by microarray hybridization, and 34 miRNAs showed strong positive signals. Five of these miRNAs were selected for further investigation by real-time RT-PCR. The statistical analyses indicated that three miRNAs exhibited significant differential expression (p<0.05) during porcine muscle development from 65 to 90 days of gestation, e.g., miR-24 and miR-424 were down-regulated while miR-133a was up-regulated. Multi-tissue RT-PCR was performed to detect the expression patterns of the five miRNA precursors. The results showed that most of these precursor miRNAs were ubiquitously expressed in different porcine tissues.

• Asian-Australasian Journal of Animal Sciences
• /
• v.31 no.1
• /
• pp.3-12
• /
• 2018

Objective: DNA methylation plays a major role in regulating the expression of genes related to traits of economic interest (e.g., weight gain) in livestock animals. This study characterized and investigated the functional inferences of genome-wide DNA methylome in the loin (longissimus dorsi) muscle (LDM) of swine. Methods: A total of 8.99 Gb methylated DNA immunoprecipitation sequence data were obtained from LDM samples of eight Duroc pigs (four pairs of littermates). The reference pig genome was annotated with 78.5% of the raw reads. A total of 33,506 putative methylated regions (PMR) were identified from methylated regions that overlapped at least two samples. Results: Of these, only 3.1% were commonly observed in all eight samples. DNA methylation patterns between two littermates were as diverse as between unrelated individuals (p = 0.47), indicating that maternal genetic effects have little influence on the variation in DNA methylation of porcine LDM. The highest density of PMR was observed on chromosome 10. A major proportion (47.7%) of PMR was present in the repeat regions, followed by introns (21.5%). The highest conservation of PMR was found in CpG islands (12.1%). These results show an important role for DNA methylation in species- and tissue-specific regulation of gene expression. PMR were also significantly related to muscular cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism. Conclusion: This study indicated the biased distribution and functional role of DNA methylation in gene expression of porcine LDM. DNA methylation was related to cell development, cell-cell communication, cellular integrity and transport, and nutrient metabolism (e.g., insulin signaling pathways). Nutritional and environmental management may have a significant impact on the variation in DNA methylation of porcine LDM.