• Journal of Embryo Transfer
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• v.21 no.3
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• pp.263-271
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• 2006

The purpose of this study was undertaken to compare ability of frozen-thawed sperm characteristics between two strains (miniature pig and Duroc). The semen was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle. The semen was diluted with same volume extender and added to LEY solution for freezing. The diluted semen was placed in 0.5 ml straws, and freezing was initiated by exposing the straws to liquid nitrogen ($LN_2$) vapours for 10 min before placing them into $LN_2$ for cryopreservation. The frozen-semen straw were thawed at 20, 37 and $50^{\circ}C$ for 1 min, 45 sec and 10 sec within water-bath. The semen sample were evaluated at 0, 3, 6, 9, and 12 h after incubation at $37^{\circ}C$ for analysis of sperm ability. Abnormality of spermatozoa in miniature pig was significantly (p<0.05) higher than that in Duroc at 0, 9 and 12 h of post-thawing incubation after frozen-thawing. The percentage of F-patterned spermatozoa in miniature pig was significantly (p<0.05) lower, while the percentage of AR (acrosome reacted spermatozoa) pattern was higher in the miniature than in the Duroc. On the other hand, there was no significant difference in the viability of spermatozoa thawed at different temperature ($20^{\circ}C\;and\;37^{\circ}C$) between two species, but the viability in miniature pig was higher (p<0.05) than in Duroc when sperm was thawed at $50^{\circ}C$. In conclusion, this study suggest that suitable freezing method for miniature pig semen is required for increasing post-thawing viability and fertilization capacity.

• Journal of Food Hygiene and Safety
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• v.27 no.3
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• pp.317-324
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• 2012

In order to determine an authenticity of food ingredient, we used DNA barcode method by universal primers. For identification of animal food ingredients, LCO1490/HCO2198 and VF2/FISH R2 designed for amplifying cytochrome c oxidase subunit1 (CO1) region and L14724/H15915 for cytochrome b (cyt b) region on mitochondrial DNA were used. Livestock (cow, pig, goat, sheep, a horse and deer) was amplified by LCO1490/HCO 2198, VF2/FISH R2 and L14724/H15915 primers. Poultry (chicken, duck, turkey and ostrich) was amplified by LCO1490/HCO 2198 and VF2/FISH R2 primers. But, Fishes (walleye pollack, herring, codfish, blue codfish, trout, tuna and rockfish) were only amplified by VF2/FISH R2 primers. For plant food ingredients, 3 types of primers (trnH/psbA, rpoB 1F/4R and rbcL 1F/724R) have been used an intergenic spacer, a RNA polymerase beta subunit and a ribulose bisphosphate carboxylase region on plastid, respectively. Garlic, onion, radish, green tea and spinach were amplified by trnH/psbA, rpoB 1F/4R and rbcL 1F/724R. The PCR product sizes were same by rpoB 1F/4R and rbcL 1F/724R but, the PCR product size using trnH/psbA primer was different with others for plants each. We established PCR condition and universal primer selection for 17 item's raw materials for foods and determine base sequences aim to PCR products in this study. This study can apply to determine an authenticity of foods through making an comparison between databases and base sequences in gene bank. Therefore, DNA barcode method using universal primers can be a useful for species identification techniques not only raw materials but also processed foods that are difficult to analyze by chemical analysis.

• Journal of Food Hygiene and Safety
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• v.25 no.4
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• pp.402-409
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• 2010

The study was conducted to estimate the contents of heavy metal in commercial herbal medicines (1047 samples of 132 species) which were collected from markets in Seoul and to analyze the contents of heavy metals of herbal medicines by classifying them by parts. The samples were digested using microwave method. The contents of heavy metal (Pb, Cd, and As) and Hg were determined using Inductively coupled plasma-Mass spectrometer (ICP/MS). And the contents of Hg were obtained by Mercury analyzer. The average values of heavy metal in herbal medicines were as follows [mean (minimum-maximum), mg/kg]; Pb 0.870 (ND-69.200), As 0.148 (ND-2.965), Cd 0.092 (ND-2.010), and Hg 0.007 (ND-0.B7). And the average values of heavy metal by parts in herbal medicines were as follows [mean (minimum-maximum), mg/kg]; Ramulus 2.046 (0.065-4.474), Herba 1.886 (0.048-10.404), Flos 1.874 (0.052-5.393), Cortex 1.377 (0.011-4.837), Radix 1.165 (0.012-70.111), Rhizoma 1.116 (0.016-5.490, Fructus 0.838 (0.017-4.527), Perithecium 0.729 (0.013-4.953), Semen 0.646 (0.006-4.416). The average values of heavy metal of imported herbal medicines except Radix were higher than domestic ones. By decoction of herbal medicines exceeding the tolerances, average intake rates of Pb, As, Cd and Hg were obtained as 6.1%, 40.3%, 4.7%, and 2.2%, respectively.

• Journal of Food Hygiene and Safety
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• v.33 no.5
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• pp.404-411
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• 2018

To evaluate bacteriological and toxicological safety hygienic indicator bacterium and paralytic and diarrhetic shellfish toxins in the shellfish produced in Hansan Geojeman 2013-2017 were investigated. Fecal coliforms were < 18~330 MPN/100 g in 404 oyster samples. But all samples tested, did not exceed 230 E. coli MPN/100 g. Geometric mean of E. coli for oyster samples collected during major shellfish production period was 24.3 MPN/100 g, considerde stable results. Bacteriological quality of oysters collected from Hansan Geojeman meets the standard value based on shellfish hygiene of the Food Sanitation Act of Korea and also meets Grade A, according to classification of shellfish harvesting areas of the European Union. For toxicological evaluation of Hansan Geojeman, 532 oyster samples and 268 mussel samples as an indicator, were analyzed. Paralytic shellfish toxins were detected in the range of 0.42~2.29 mg/kg in eight mussel samples, and exceeded criteria in three samples from early to late April 2013. Diarrhetic shellfish toxin was detected in three of 120 samples, but it was revealed to be under regulation value (0.16 mg Okadaic Acid equ./kg). As a result of toxicological evaluation, paralytic and diarrhetic shellfish toxins were not detected in oyster samples, but it was found that mussel as an indicator species, exceeded the threshold value of paralytic shellfish toxin. Accordingly, sanitary surveys were continuously requested for food safety management of shellfish.

• Journal of Life Science
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• v.21 no.3
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• pp.385-392
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• 2011

The otter (Lutra lutra) in Korea is classified as a first grade endangered species and is managed under state control. We performed a phylogenetic analysis of the otter that inhabits the Changnyeong, Jinju, and Geoje areas in Gyeongsangnamdo, Korea using mtDNA and microsatellite (MS) markers. As a result of the analysis using the 676-bp D-loop sequence of mtDNA, six haplotypes were estimated from five single nucleotide polymorphisms. The genetic distance between the Jinju and Geoje areas was greater than distances within the areas, and the distance between Jinju and Geoje was especially clear. From the phylogenetic tree estimated using the Bayesian Markov chain Monte Carlo analysis by the MrBays program, two subgroups, one containing samples from Jinju and the other containing samples from the Changnyeong and Geoje areas were clearly identified. The result of a parsimonious median-joining network analysis also showed two clear subgroups, supporting the result of the phylogenetic analysis. On the other hand, in the consensus tree estimated using the genetic distances estimated from the genotypes of 13 MS markers, there were clear two subgroups, one containing samples from the Jinju, Geoje and Changnyeong areas and the other containing samples from only the Jinju area. The samples were not identically classified into each subgroup defined by mtDNA and MS markers. It could be inferred that the differential classification of samples by the two different marker systems was because of the different characteristics of the marker systems used, that is, the mtDNA was for detecting maternal lineage and the MS markers were for estimating autosomal genetic distances. Nonetheless, the results from the two marker systems showed that there has been a progressive genetic fixation according to the habitats of the otters. Further analyses using not only newly developed MS markers that will possess more analytical power but also the whole mtDNA are needed. Expansion of the phylogenetic analysis using otter samples collected from the major habitats in Korea should be helpful in scientifically and efficiently maintaining and preserving them.

• Journal of Life Science
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• v.21 no.7
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• pp.961-968
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• 2011

The formation of reactive lipid aldehydes, 4-hydroxynonenal (HNE) is shown to be derived from fatty acid hydroperoxides through the oxidative process. Among its known effects in cytotoxicity, HNE has been implicated in apoptotic cell death. To delineate its putative role as a potential mediator, we investigated the mechanism by which HNE induces apoptosis of endothelial cells (ECs). The anti-proliferative effects of HNE were tested through MTT assay after exposure to various concentrations ($5\sim15\;{\mu}M$) of HNE. We observed apoptotic bodies with propidium iodide staining, and measured the HNE induction of endothelial apoptosis by flow cytometry assay. We observed that cells exposed to HNE for 24 hr resulted in increased poly(ADP-ribose) polymerase cleavage and up-regulation of Bax. Data on the HNE action strongly indicated the involvement of reactive species, namely, intracellular ROS, nitrite, and peroxynitrite. To obtain evidence on the implication of ROS and peroxynitrite in HNE-induced apoptosis, a ROS scavenger, N-acetylcysteine (NAC), and a peroxynitrite scavenger, penicillamine, were tested. Results clearly indicate that the induction of apoptosis by HNE was effectively inhibited by NAC and penicillamine. Based on the present data, we conclude that the endothelial apoptosis induced by HNE involves both ROS generation and peroxynitrite activity. Our new data could lead to a redefinition of HNE action on apoptosis in ECs.

• Journal of Life Science
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• v.27 no.5
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• pp.509-516
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• 2017

Perilla frutescens (L.) Britton var. sprouts (PFS) is a plant of the labiatae family. The purpose of this work was to assess the preventive effects of PFS ethanolic extracts (PFSEs) on cytokine-induced ${\beta}$-cell damage. Cytokines, which are released by the infiltration of inflammatory cells around the pancreatic islets, are involved in the pathogenesis of type 1 diabetes mellitus. The combination of interleukin-$1{\beta}$ (IL-1), interferon-${\gamma}$ (IFN-${\gamma}$), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) induced formation of reactive oxygen species (ROS). Accumulation of intracellular ROS led to ${\beta}$-cell dysfunction and apoptosis. PFSEs possess antioxidant activity and thus lead to downregulation of ROS generation. Cytokines decrease cell viability, stimulate the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and induce the production of nitric oxide (NO). PFSEs prevented cytokine-induced cell viability in a dose-dependent manner. Incubation with PFSE resulted in significant reduction in cytokine-induced NO production that correlated with reduced levels of the iNOS and COX-2 protein expression. Furthermore, PFSE significantly decreased the activation of nuclear factor ${\kappa}B$ (NF-${\kappa}B$) by inhibition of $I{\kappa}B{\alpha}$ phosphorylation in RINm5F cells. In summary, our results suggest that the protective effects of PFSE might serve to counteract cytokine-induced ${\beta}$-cell destruction. Findings indicate that consumption of Perilla frutescens (L.) Britton var. sprouts alleviates hyperglycemia-mediated oxidative stress and pro-inflammatory cytokine-induced ${\beta}$-cell damage and thus has beneficial anti-diabetic effects.

• Journal of Life Science
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• v.27 no.7
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• pp.739-745
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• 2017

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

• Journal of Life Science
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• v.27 no.10
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• pp.1152-1160
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• 2017

A unicellular green alga was axenically isolated from a tidal pool on Ulleung-do, Korea. Morphological, molecular, and biochemical analyses revealed that the isolate belonged to Auxenochlorella protothecoides. The current study is the first record of this species in Korea. The microalgal strain was named as A. protothecoides MM0011 and its growth, lipid and pigment compositions, and biomass properties were investigated. The strain is able to thrive in a wide range of temperatures ($5{\sim}35^{\circ}C$) and to withstand up to 1.5 M NaCl. The results of GC/MS analysis showed that the isolate was rich in nutritionally important polyunsaturated fatty acids (PUFAs). Its major fatty acids were linoleic acid (27.6%) and ${\alpha}-linolenic$ acid (39.6%). Thus, this indigenous microalga has potential as an alternative source of ${\omega}3$ and ${\omega}6$ PUFAs, which currently come from fish and plant oils. Also, the HPLC analysis revealed that the value-added antioxidant, lutein, was biosynthesized as the accessory pigments by the microalga. A proximate analysis showed that the volatile matter content was 85.6% and an ultimate analysis indicated that the gross calorific value was $20.3MJ\;kg^{-1}$. Since 40.5% of total nitrogen and 27.9% of total phosphorus were removed from the medium, respectively, it also has potential as a feedstock for biofuel applications which could be coupled to wastewater treatment. In addition, the biomass may also serve as an excellent animal feed because of its high protein content (51.4%). Therefore, A. protothecoides MM0011 shows promise for application in production of microalgae-based biochemicals and as a biomass feedstock.

• Tuberculosis and Respiratory Diseases
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• v.39 no.4
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• pp.304-309
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• 1992

Background: The alveolar macrophage may metabolize arachidonic acid through cyclooxygenase- and lipoxygenase- catalyzed pathways to produce a variety of metabolites of arachidonic acid. The production of these metabolites of arachidonic acid may enhance the defensive ability of the challenged lung. However, continued stimulation with the consequent production of proinflammtory metabolites of arachidonic acid, may ultimately enhance the disease process by contributing to chronic bronchoconstriction, fibrosis, and the persistent release of toxic oxygen species. Silicosis is an example of a disease process resulting from chronic exposure of the lung to foreign particles. This study was carried out to evaluate the changes of arachidonic acid metabolites from macrophages in experimental silicosis. Methods: We measured $PGE_2$, and $LTB_4$ in cultured macrophages taken from rats by radioimmunoassay at 24 and 48 hours after stimulation by silica dust, natural carbon dust, lipopolysaccharide, calcium ionophore (A23187) and medium (RPMI) as a control. For the experimental silicosis, 50 mg silica in 0.5 ml saline was administered intratracheally into the rat and grown to 20 weeks and measured $PGE_2$, and $LTB_4$ in the cultured macrophages lavaged from that rat. The used stimulants were the same as above. Results: 1) The amount of $PGE_2$ in the cultred macrophages from normal rat was significantly decreased in the group which was stimulated with silica dust for 48 hours compare with control non-stimulated group. 2) In the experimental silicosis group, $PGE_2$, release in cultured macrophages after 48 hours incubation with silica and natural carbon dust tended to be lower than those of non-stimulated group. 3) There were marked changes of $LTB_4$ in the groups of normal rats which were incubated with silica for 24, 48 hours and natural carbon for 48 hours compared with non-stimulated group. 4) In the experimental silicosis group, the release of $LTB_4$ was significantly increased macrophages cultured with silica and natural carbon dust after 24 and 48 hours incubation compared with non-stimulated group. Conclusion: The results of these studies suggest that the in vitro exposure of rat alveolar macrophge to silica and coal dust results in an alteration in alveolar macrophage metabolism of arachidonic acid that may promote an inflammatory reaction in lung tissue.