• International Journal of Industrial Entomology and Biomaterials
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• v.33 no.2
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• pp.113-120
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• 2016

Bombyx mandarina is widely accepted as ancestor of B. mori. Silkworms are served as well-characterized models for understanding the mechanism for the genetic regulation of development. In this study, we performed RNA-Seq analysis to examine tissue-expression of gloverin isoforms of the silk-gland, mid-gut, and fat body in B. mandarina. BLAST analysis revealed that four gloverin isoform gene sequences of B. mandarina were highly similar to B. mori. To identify the difference between two species, the expression profile of gloverin was measured by semi- RT-PCR analysis. The specific expression of gloverin isoform genes was observed mainly in the fat body from B. mori but not B. mandarina. However, all of tissues in the wild-type silkworm could induce the upregulation of compared with the B. mori. To validate the sudden increase in gloverin gene expression in the mid-gut tissue of B. mandarina, we were using qRT-PCR. Relative mRNA expression rate of gloverin at the wild-type silkworm was much higher than domestic silkworm. Comparative genomics between domesticated and wild silkworms showed different tissue-expression levels in some of immune related genes. These results are suggesting a trend toward decreasing immunity related genes expression during domestication. Further studies are needed to elucidate the silkworm domestication and an invaluable resource for wild silkworm genomics research.

• KSBB Journal
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• v.17 no.3
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• pp.283-289
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• 2002

The present study was aimed at investigating the metabolic regulation of insulin-like growth factor-I(IGF-I) expression in fasting animals. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA from control, 4d-fasting, and 2d-fasting-refed rats. The levels of IGF-I transcripts were reduced in 4d-fasting than in control by decreasing its transcriptional rate, which was measured through nuclear nun-on assay. DNase I footprinting, which was performed using nuclear extracts from fasting rat, demonstrated protein binding to a sequence that extended from +179 to +210 (termed region B). These data suggest that the expression of IGF-I is transcriptionally regulated through DNA-liver enriched protein binding in a sequence which is located downstream from major transcription initiation site of IGF-I gene.

• Development and Reproduction
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• v.21 no.3
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• pp.317-325
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• 2017

Direct communication between neighboring cells through connexin (Cx)-based gap junction is a crucial biological manner to regulate functions of a tissue consisting of multi-cell types. The present research evaluated expressional changes of Cx isoforms in the caput epididymis of adult rat exposed to estradiol benzoate (EB) or flutamide (Flu) at the early postnatal age. A single subcutaneous administration of EB at a low-dose [$0.015{\mu}g/kg$ body weight (BW)] or a high-dose ($1.5{\mu}g/kg\;BW$) or Flu at a low-dose ($500{\mu}g/kg\;BW$) or a high-dose (5 mg/kg BW) was performed to an animal at 1 week of age. Quantitative real-time PCR analysis was employed to determine expressional changes of Cx isoforms. The transcript levels of Cxs30.3 and 37 were decreased by a low-dose EB treatment, while decreases of Cxs31, 31.1, 32, 40, and 45 transcript levels were observed with a low-dose EB treatment. The treatment of a high-dose EB resulted in expressional reduction of Cxs30.3, 31, 31.1, 37, 40, 43, and 45. The Flu treatment at a low dose caused increases of Cxs26, 37, and 40 transcript levels but decreases of Cxs31.1, 43, and 45 transcript levels. Increases of Cxs30.3, 31, 37, and 40 mRNA amounts were induced by a high-dose Flu treatment. However, exposure to a high-dose Flu produced expressional decreases of Cxs31.1, 32, and 43 in the adult caput epididymis. These observations suggest that exposure to EB or Flu at the neonatal period could lead to aberrant expression of Cx isoforms in the adult caput epididymis.

• Biomedical Science Letters
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• v.10 no.1
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• pp.23-29
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• 2004

Nebulin is a (Mr 600∼900 kDa) large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Cardiac muscles of higher vertebrates have been shown earlier to lack nebulin. Recently, full-length nebulin mRNA transcripts have been detected in heart muscle, but at lower levels than in skeletal muscle. Nebulin expression also was detected in the kidney, eye, and otic canal, suggesting that nebulin isoforms may also be expressed in these organs. We have searched for nebulin isoforms in brain of human using PCR and Northern blot. Here, we provide evidence that nebulin mRNA transcripts are expressed in brain. Seven nebulin isoforms (B, C, D, E, F, G and H form) are obtained in human skeletal muscle and four isoforms (B, C, G and H form) in human brain cDNA library. We cloned the 1.3 kb of nebulin fragment from human adult brain library by PCR. The identity of the PCR product was confirmed by sequence analysis. The partial brain nebulin sequence was 99% identical to the skeletal muscle cDNA as determined by Blast alignment. It contains two simple-repeats HR1, HR2 and linker-repeats exon l35∼143 except exon 140. It was different from skeletal muscle B form, which contain HR1 and HR8. These data suggest that nebulin isoform diversity occurs even more extensively than previously known, likely contributing to the distinct thin filament architecture of different striated muscles.

• Biomedical Science Letters
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• v.14 no.2
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• pp.69-76
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• 2008

${\gamma}$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system, and its actions are mediated by subtypes of GABA receptors named as $GABA_A$, $GABA_B,\;and\;GABA_C,\;GABA_A$, receptor consisting of ${\alpha},\;{\beta},\;{\gamma}\;and\;{\delta}$ subunits is a heterooligomeric ligand-gated chloride channel. This study was performed to investigate regulation of $GABA_A$ receptor by protein kinase C(PKC). Ion currents were recorded using gramicidine-perforated patch and whole cell patch clamp. mRNA encoding the subunits of PKC expressed in major pelvic ganglion (MPG) neurons was detected by using RT-PCR. The GABA-induced inward current was increased by PKC activators and decreased by PKC inhibitors, respectively. These effects were not associated with intracellular $Ca^{2+}$ and GAG (1-oleoyl-2-acetyl-sn-glycerol), a membrane permeable diacylglycerol (DAG) analogue. These results mean that the subfamily of PKC participating in activation of $GABA_A$ receptor would be an atypical PKC (aPKC). Among theses, ${\xi}$ isoform of aPKC was detected by RT-PCR. Taking together, we suggest that excitable $GABA_A$ receptor in sympathetic MPG neuron seemed to be regulated by aPKC, particular in ${\xi}$ isoform. The regulatory roles of PKC on excitatory $GABA_A$ receptors in sympathetic neurons of MPG may be an important factor to control the functional activity of various pelvic organs such as bowel movement, micturition and erection.

• Biomedical Science Letters
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• v.2 no.1
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• pp.21-30
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• 1996

MAP kinases are a family of serine/threonine specific protein kinases becoming activated in response to different proliferative stimuli by phosphorylation at both threonine and tyrosine residue. Present study shows that MAP kinase was purified from P388 murine leukema cells by SP sephadex C-50, phenyl superose and Mono Q column chromatography and identified with anti-ERKl antibody by western blotting. Immnublotting analysis to the crude extract of P388 cell lysate shows 44 kD and other minor bands but partial purified fraction eluted from phenyl supherose column have 44kD and 66 kD isoform. Subcloned GST-fusion protein from N-terminal of $p56^{kk}$ was tested as a substrate for MAP kinase phosphorylation. It was showed that the wild type and mutant forms(S42A) were fully phosporylated by purified MAP kinase fraction as com-pare with the other mutant form(S59A). This finding suggest that those GST-fusion proteins may be used as substrate for the in vitro test of MAP kinase.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.10
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• pp.1612-1617
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• 2007

In this study, we have compared the skeletal muscle proteome at various stages of porcine postnatal development. Korean native pigs were divided into five postnatal stages of 30, 70, 130, 170 and 300 d and their loin muscles were analyzed for muscle proteome by using two-dimensional electrophoresis and mass spectrometry. We found 5 proteins showing a consistent pattern during skeletal muscle growth. Four proteins were identified as myosin light chain 1 slow-twitch (MLC1sa) isoform, troponin T, triosephosphate isomerase (TIP) and DJ-1 protein. The remaining protein was not identified. Two muscle fiber proteins of MLC1sa isoform and troponin T showed a high expression level at an early postnatal stage and then their levels were decreased markedly during growth stages. On the other hand, the expression of TIP and DJ-1 protein, which are well known as catalysis enzyme and antioxidant-related protein, respectively, were linearly increased during growth stages. Thus, the stage-related muscle proteins may be useful as parameters for understanding the developmental characteristics of biochemical and physiological properties in Korean native pig skeletal muscle.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.3
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• pp.161-165
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• 2004

Exogenous carbon monoxide (0.2%) increases L-type calcium $(Ca^{2+})$ current in human jejunal circular smooth muscle cells. The stimulatory effect of carbon monoxide (CO) on L-type $Ca^{2+}$ current is inhibited by pre-application of L-NNA, a classical competitive inhibitor of nitric oxide synthase (NOS) with no significant isoform selectivity (Lim, 2003). In the present study, we investigated which isoform of NOS affected CO induced stimulation of L-type $Ca^{2+}$ current in human jejunal circular smooth muscle cells. Cells were voltage clamped by whole-cell mode patch clamp technique, and membrane currents were recorded with 10 mM barium as the charge carrier. Before the addition of CO, cells were pretreated with each inhibitor of three NOS isoforms for 15 minutes. CO-stimulating effect on L-type $Ca^{2+}$ current was partially blocked by N-(3-(Amino-methyl) benzyl) acetamidine 2HCl (1400W, an iNOS inhibitor). On the other hand, 3-bromo-7-nitroindazole (BNI, a nNOS inhibitor) or $N^5-(1-Iminoethyl)-L-ornithine$ dihydrochloride (L-NIO, an eNOS inhibitor) completely blocked the CO effect. These data suggest that low dose of exogenous CO may stimulate all NOS isoforms to increase L-type $Ca^{2+}$ channel through nitric oxide (NO) pathway in human jejunal circular smooth muscle cells.

• Food Science of Animal Resources
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• v.31 no.4
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• pp.506-512
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• 2011

This study was conducted to validate the theoretical feasibility of a technique to identify biomarkers in Korean native black pig (KNP) and a commercial Landrace breed. Using two-dimensional electrophoresis, we found six proteins (NADH dehydrogenase Fe-S protein 1, an unnamed protein product, similar to T-complex protein 1, annexin V = CaBP33 isoform, fatty acid-binding protein, and catechol O-methyltransferase), which appeared in KNP alone. We raised polyclonal antibodies (used as the primary antibody) for Western blotting to confirm the characteristics of the six KNP proteins. As a result, catechol O-methyltransferase, annexin V = CaBP33 isoform, and the unnamed protein product presented thicker bands in KNP than those in Landrace. Moreover, catechol O-methyltransferase was shown to be more feasible as a biomarker for KNP. However, cross-reactivity was observed with the polyclonal antibodies for KNP and the other three proteins (NADH dehydrogenase, a protein similar to T-complex protein 1, and fatty acid-binding protein). This study only showed limited results from a limited number of animals; however, our research suggests possibilities for future studies.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.3
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• pp.1163-1168
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• 2016

Cholangiocarcinoma (CCA) is the bile duct cancer which constitutes one of the important public health problems in Thailand with high mortality rate, especially in the Opisthorchis viverrini (a parasite risk factor for CCA) endemic area of the northeastern region of the country. This study aimed to identify potential biomarkers from the plasma peptidome by CCA patients. Peptides were isolated using 10 kDa cut-off filter column and the flow-through was then used as a peptidome for LC-MS/MS analysis. A total of 209 peptides were obtained. Among these, 15 peptides were concerned with signaling pathways and 12 related to metabolic, regulatory, and biosynthesis of secondary metabolite pathways. Five exclusive peptides were identified as potential biomarkers, i.e. ETS domain-containing transcription factor ERF (P50548), KIAA0220 (Q92617), phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit beta isoform isoform 1 (P42338), LP2209 (Q6XYC0), and casein kinase II subunit alpha (P19784). Three of these biomarkers are signaling related molecules. A combination of these biomarkers for CCA diagnosis is proposed.