• Development and Reproduction
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• v.5 no.2
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• pp.115-122
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• 2001

Production of a mature oocyte is a complex process that requires the close association between oocyte and follicular cells. The present study was conducted to investigate the difference between oocytes with and without close junctional communication with cumulus cells and the involvement of two connexins(CXs) in the interactions. Follicles at different sizes(small: 200～400 ㎛; large:>450 ㎛) were mechanically isolated from PMSG-primed mouse ovaries, and punctured to get cumulus-oocyte complex(COC). Oocytes were released themselves(denuded), with partially attached(partial), or with tightly attached(intact) cumulus cells. Maturation and fertilization capacity of the COCs were measured. Expression of CX 37 and CX 43 was examined by RT-PCR and in situ hybridization. The ratio between intact COC and denude/partial oocytes was 30％(SI) and 70％(SPD) in small follicles, while 55％(LI) and 45％(LPD) in large follicles, respectively. Maturation and fertilization rates of the released oocytes were similar among SI, LPD, LI groups, but those were significantly lower in SPD oocytes. In oocytes, CX 37 was the major CX and CX 43 was not expressed, whereas in the cumulus cells, CX 43 was the major, and CX 37 was the next. Results of the present study suggest that 1) immature oocytes from small follicles with intact cell-to-cell communication with cumulus have the similar quality to that of the oocytes from larger follicles, 2) gap junction between oocyte and cumulus cells may be the heterotypic channel, and 3) we could not explain the difference in the cell-to-cell communication between intact and partial/denuded COCs through the expression of the two CXs.

• Food Science of Animal Resources
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• v.34 no.5
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• pp.656-664
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• 2014

The purpose of this study was to analyze myosin heavy chain (MHC) isoforms in bovine longissimus thoracis (LT) muscle by liquid chromatography (LC) and mass spectrometry (MS). LT muscles taken from Hanwoo (Korean native cattle) steer (n=3) used to separate myosin bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The peptide queries were obtained from the myosin bands by LC-MS/MS analysis following in-gel digestion with trypsin. A total of 33 and 43 queries were identified as common and unique peptides, respectively, of MHC isoforms (individual ions scores >43 indicate identity or extensive homology, p<0.05). MHC-1 (IIx), -2 (IIa), -4 (IIb), and -7 (slow/I) were identified based on the Mowse score (5118, 3951, 2526, and 2541 for MHC-1, -2, -4, and -7, respectively). However, more analysis is needed to confirm the expression of MHC-4 in bovine LT muscle because any query identified as a unique peptide of MHC-4 was not found. The queries that were identified as unique peptides could be used as peptide markers to confirm MHC-1 (14 queries), -2 (8 queries), and -7 (21 queries) in bovine LT muscle; no query identified as a unique peptide of MHC-4 was found. LC-MS/MS analysis is a useful approach to study MHC isoforms at the protein level.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.3
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• pp.350-355
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• 2009

Arachidonic acid (AA) is a polyunsaturated fatty acid that is a normal constituent of membrane lipids in animal cells. In addition to its role as a precursor of prostaglandins, AA itself may play an important role in the regulation of cell function. The effect of AA on functions of granulosa cells was investigated in pre-hierarchical small yellow follicles of laying hens. Immuno-cytochemical staining showed that AA ($10^{-7}-10^{-5}$ M) increased the expression of the extracellular matrix glycoprotein laminin, gap junction connexin 43 and protein kinase C (PKC). Therefore, mediated by the PKC signal pathway, AA may regulate the intercellular communication of granulosa cells and follicular development by increasing the expression of laminin and connexin.

• Applied Biological Chemistry
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• v.41 no.5
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• pp.349-354
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• 1998

The protease produced by Bacillus subtilis YG-95 was purified by precipitating with ammonium sulfate, DEAE-sepharose 6B and Sephadex G-100 column chromatogtaphies and its purified enzymological characteritics were investigated. The molecular weight of purified protease was estimated about 43kilodalton by SDS PAGE The optimum pH and temperature for the purified protease activity were pH 10.0 and $55^{\circ}C$, respectively. The enzyme was stable in broad range of pH 5.0 to 12.0. and at the below $45^{\circ}C$. The purified enzyme activty was inhibited by $Fe^{3+}$ and $Al^{3+}$. The activity was significantly inhibited more than 80% by O-Phenanthroline, PMSF and SDS. The $K_m$ value of the purified enzyme against Soy Protein Isolate as a substrate was 1.28 mg/ml.

• Environmental Mutagens and Carcinogens
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• v.23 no.2
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• pp.56-62
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• 2003

The liver progenitor cells could form a potential target cell population fore both tumor-initiating and -promoting chemicals. Induction of drug-metabolizing and antioxidant enzymes, including AhR-dependent CYP1A1, NQO-1 and AKR1C9, was detected in the rat liver epithelial WB-F344 "stem-like" cells. Additionally, WB-F344 cells express a functional, wild-type form of p53 protein, a biomarker of genotoxic events, and connexin 43, a basic structural unit of gap junctions forming an important type of intercellular communication. In this cellular model, two complementary assays have been established for detection of the modes of action associated with tumor promotion: inhibition of gap junctional intercellular communication (GJIC) and proliferative activity in confluent cells. We found that the PAHs and PCBs, which are AhR agonists, released WB-F344 cells from contact inhibition, increasing both DNA synthesis and cell numbers. Genotoxic effects of some PAHs that lead to apoptosis and cell cycle delay might interfere with the proliferative activity of PAHs. Contrary to that, the nongenotoxic low-molecular-weight PAHs and non-dioxin-like PCB congeners, abundant in the environment, did not significantly affect cell cycle and cell proliferation; however both groups of compounds inhibited GJIC in WB-F344 cells. The release from contact inhibiton by a mechanism that possibly involves the AhR activation, inhibition of GJIC and genotoxic events induced by environmental contaminants are three important modes of action that could play an important role in carcinogenic effects of toxic compounds. The relative potencies to inhibit GJIC, to induce AhR-mediated activity, and to release cells from contact inhibition were determined for a large series of PAHs and PCBs and their metabolites. In vitro bioassays based on detection of events on cellular level (deregulation of GJIC and/or proliferation) or determination of receptor-mediated activities in both ?$stem-like^{\circ}{\times}$ and hepatocyte-like liver cellular models are valuable tools for detection of modes of action of polyaromatic hydrocarbons. They may serve, together with concentration data, as a first step in their risk assessment.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.425-430
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• 2014

Foods contain various nutrients such as carbohydrates, protein, oil, vitamins, and minerals. Among them, carbohydrates, protein, and oil are the main constituents of foods. Usually, these constituents are analyzed by the Kjeldahl and Soxhlet method and so on. However, these analytical methods are complex, costly, and time-consuming. Thus, this study aimed to rapidly and effectively analyze carbohydrate, protein, and oil contents with near-infrared reflectance spectroscopy (NIRS). A total of 517 food samples were measured within the wavelength range of 400 to 2,500 nm. Exactly 412 food calibration samples and 162 validation samples were used for NIRS equation development and validation, respectively. In the NIRS equation of carbohydrates, the most accurate equation was obtained under 1, 4, 5, 1 (1st derivative, 4 nm gap, 5 points smoothing, and 1 point second smoothing) math treatment conditions using the weighted MSC (multiplicative scatter correction) scatter correction method with MPLS (modified partial least square) regression. In the case of protein and oil, the best equation were obtained under 2, 5, 5, 3 and 1, 1, 1, 1 conditions, respectively, using standard MSC and standard normal variate only scatter correction methods with MPLS regression. Calibrations of these NIRS equations showed a very high coefficient of determination in calibration ($R^2$: carbohydrates, 0.971; protein, 0.974; oil, 0.937) and low standard error of calibration (carbohydrates, 4.066; protein, 1.080; oil, 1.890). Optimal equation conditions were applied to a validation set of 162 samples. Validation results of these NIRS equations showed a very high coefficient of determination in prediction ($r^2$: carbohydrates, 0.987; protein, 0.970; oil, 0.947) and low standard error of prediction (carbohydrates, 2.515; protein, 1.144; oil, 1.370). Therefore, these NIRS equations can be applicable for determination of carbohydrates, proteins, and oil contents in various foods.

• The Journal of Korean Medicine
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• v.34 no.3
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• pp.126-142
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• 2013

Objectives: Previous reports have shown that Bogijetong-Tang (BJT) is effective in peripheral neuropathy induced by taxol and crush injury. In this study, we researched the effects of BJT on diabetic neuropathy induced by STZ in the mouse. Methods: We performed both in vitro and in vivo experiments to verify the effects of BJT on diabetic neuropathy induced by STZ in mice. Changes in axonal recovery were observed with immunofluorescence staining using NF-200, Hoechst33258, $S100{\beta}$, caspase 3 and anti-cdc2. Proliferation and degeneration of Schwann cells were investigated by immunofluorescence staining and western blot analyses. Results: BJT showed considerable effects on neurite outgrowth and axonal regeneration in diabetic neuropathy. BJT contributed to the creation of NF-200, GAP-43, Cdc2, phospho-vimentin, ${\beta}1$, active ${\beta}1$, ${\beta}3$ integrin, phospho-Erk1/2 protein. Conclusions: Through this study, we found that BJT is effective for enhanced axonal regeneration via dynamic regulation of regeneration-associated proteins. Therefore, BJT had a pharmaceutical property enhancing recovery of peripheral nerves induced by diabetic neuropathy and could be a candidate for drug development after more research.

• Journal of Sericultural and Entomological Science
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• v.40 no.1
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• pp.43-51
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• 1998

Studies were carried out to investigate phenotypic expression, mortality and biochemical analysis of haemolymph proteins of nm-d, nm-f, nm-k and nmn bib-molting mutants of the silkworm, Bombyx mori. The non-molting mutants characters were expressed in the homozygote of each mutant genes. All strains of non-molting mutants were similar with each other in physiological characteristics, but the expression varied with each strains. The larvae of nm-d, nm-i and nmn died between day 5 and day 9 after hatching without the first molt. The nm-f and nm-k mutants died between day 5 and day 16 with a slight increase of body weight and, more than 90% of the mutants larvae died before the first molt and a few of them survived to the 2nd and the 3rd instar and died. The haemolymph protein components of nm-d, nm-i, and nmn were rapidly reduced, and on the other hand those of nm-f and nm-k consistently until they died. And there were no distinguishable difference in haemolymph components of non-molting mutants, as compared to those normals.

• Biomolecules & Therapeutics
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• v.22 no.2
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• pp.114-121
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• 2014

Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.

• Journal of fish pathology
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• v.14 no.2
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• pp.59-64
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• 2001

A complementary DNA encoding metallothionein(MT), a heavy metal-responsive protein was cloned from a cartilaginous shark species. Scyliorhinus torazame. An expressed sequence tag(EST)from the shark liver, which showed high similarity with a MT gene, was isolated and its full-length sequence(390bp)was determined. The putative shark MT cDNA sequence contained an open reading frame consisting 68 amino acids and 182bp of 3-untranslated region including the poly (A+) signal. The deduced amino acid sequence was 41-54% identical to those of other animals including mammals and fish species. Tiger shark MT cDNA showed high conservation in the Cys regions. however, peculiarly contained not only additional five amino acids just prior to the conserved beta-domain but also a Ser residue at C terminal, which has not been seen in other MT sequences.