• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.194-194
• /
• 2017

Camelina (Camelina sativa L.) is a potential bio-energy crop that has short life cycle about 90 days and contains high amount of unsaturated fatty acid which is adequate to bio-diesel production. Enhancing environmental stress tolerance is a main issue to increase not only crop productivity but also big mass production. CsRCI2s (Rare Cold Inducible 2) are cold and salt stress related protein that localized at plasma membrane (PM) and assume to be membrane potential regulation factor. These proteins can be divide into C-terminal tail (CsRCI2D/E/F/G) or no-tail group (CsRCI2A/B/C/H). However, function of CsRCI2s are less understood. In this study, physiological responses and functional characterization of CsRCI2s of Camelina under salt stress were analyzed. Full-length CsRCI2s (A/B/E/F) and CsPIP2;1 sequences were confirmed from Camelina genome browser. Physiological investigations were carried out using one- or four-week-old Camelina under NaCl stress with dose and time dependent manner. Transcriptional changes of CsRCI2A/B/E/F and CsPIP2;1 were determined using qRT-PCR in one-week-old Camelina seedlings treated with NaCl. Translational changes of CsRCI2E and CsPIP2;1 were confirmed with western-blot using the antibodies. Water transport activity and membrane potential measurement were observed by cRNA injected Xenopus laevis oocyte. As results, root growth rate and physiological parameters such as stomatal conductance, chlorophyll fluorescence, and electrolyte leakage showed significant inhibition in 100 and 150 mM NaCl. Transcriptional level of CsPIP2;1 did not changed but CsRCI2s were significantly increased by NaCl concentration, however, no-tail type CsRCI2A and CsRCI2B increased earlier than tail type CsRCI2E and CsRCI2F. Translational changes of CsPIP2;1 was constitutively maintained under NaCl stress. But, accumulation of CsRCI2E significantly increased by NaCl stress. CsPIP2;1 and CsRCI2A/B/E/F co-expressed Xenopus laevis oocyte showed decreased water transport activity as 61.84, 60.30, 62.91 and 76.51 % at CsRCI2A, CsRCI2B, CsRCI2E and CsRCI2F co-expression when compare with single expression of CsPIP2;1, respectively. Moreover, oocyte membrane potential was significantly hyperpolarized by co-expression of CsRCI2s. However, higher hyperpolarized level was observed in tail-type CsRCI2E and CsRCI2F than others, especially, CsRCI2E showed highest level. It means transport of $Na^+$ ion into cell is negatively regulated by expression of CsRCI2s, and, function of C-terminal tail is might be related with $Na^+$ ion influx. In conclusion, accumulation of NaCl-induced CsRCI2 proteins are related with $Na^+$ ion exclusion and prevent water loss by CsPIP2;1 under NaCl stress.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
• /
• v.16 no.4
• /
• pp.196-205
• /
• 2011

To investigate the contribution of macroalgae to biogeochemical nutrients and carbon cycles, we measured the uptake rates of nutrients and $CO_2$ and characteristics of fluorescence of Saccharina japonica (Laminaria japonica Areschoug) using an incubation method in an acrylic chamber. From January to May 2011, S.japonica was sampled at Ilkwang, one of well-known macroalgae culture sites around Korea and ranged 46~288 cm long and 4.8~22.0 cm wide of whole thallus. The production rate of dissolved oxygen by S. japonica (n=25) was about $6.9{\pm}5.8{\mu}mol\;g^{-1}$ fresh weight(FW) $h^{-1}$. The uptake rate of total dissolved inorganic carbon ($TCO_2$), calculated by total alkalinity and pH, was $8.9{\pm}7.9{\mu}mol\;g^{-1}\;FW\;h^{-1}$. Mean nutrients uptake were $175.6{\pm}161.1\;nmol\;N\;g^{-1}\;FW\;h^{-1}$ and $12.7{\pm}10.1\;nmol\;P\;g^{-1}\;FW\;h^{-1}$. There were logarithmic relationships between thallus length and uptake rates of nutrients and $CO_2$, which suggested that younger specimens (<100-150 cm) were much more efficient at nutrients and $CO_2$ uptake than old specimens > 150 cm. There was a positive linear correlation ($r^2$=9.4) existed between the dissolved oxygen production rate and the $TCO_2$ uptake rate, suggesting that these two factors may serve as good indicators of S. japonica photosynthesis. There was also positive linear relationship between maximal quantum yield ($F_v/F_m$) and production/uptake rates of dissolved oxygen, $TCO_2$ and phosphate, suggested that $F_v/F_m$ could be used as a good indicator of photosynthetic ability and $TCO_2$ consumption of macroalgae. Maximum relative electron transport rate ($rETR_{max}$) of S. japonica increased as thallus grew and was high in distal part of thallus which may be resulted from the increase of photosynthetic cell density per area. The annual $TCO_2$ uptake by S. japonica in Gijang area was estimated about $1.0\sim1.7{\times}10^3C$ ton, which was about 0.02-0.03% of carbon dioxide emission in Busan City. Thus, more research should be focused on macroalgae-based biogeochemical cycles to evaluate the roles and contributions of macroalgae to the global carbon cycle.

• Korean Journal of Ichthyology
• /
• v.19 no.2
• /
• pp.107-111
• /
• 2007

Sex differentiation of the brown croaker Miichthys miiuy (Basilewsky) is described from hatching to the 120th day post-hatching (dph) (water temperature $24^{\circ}C$). Primordial germ cells (PGCs) were observed on the 20th dph (10.4 mm total length (TL), 0.14 g body weight (BW), and began to protrude into the peritoneal cavity from the 40th dph (19.4 mm TL, 0.39 g BW). On the 65th dph (31.3 mm TL, 0.93 g BW, $1,560D^{\circ}$ (degree-days)), initial ovarian differentiation was identified by the PGCs with condensed chromatin, and their transformation into meiotic oocytes. By the 120th dph (4.60 mm TL, 1.38 g BW, $2,880D^{\circ}$), the oocytes were in the perinucleolus stage and had increased from 20 to $40{\mu}m$ in diameter. While ovaries gradually grew after sex was differentiated, testes continued to multiply from the 65th dph. On the 80th dph (37.9 mm TL, 1.39 g BW, $1,920D^{\circ}$), the beginning of testis lobule formation was indicated by the occurrence of spermatogonial cysts enveloped by somatic cells in some of the testes. On the 120th dph, the testis lobules of some of the fish contained all germ cell stages through to the spermatocytes. Therefore, the sex differentiation type of the brown croaker is identified as gonochoristic.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.3
• /
• pp.439-445
• /
• 2017

Objective: Production of alpha-1,3-galactosyltransferase (${\alpha}GT$)-deficient pigs is essential to overcome xenograft rejection in pig-to-human xenotransplantation. However, the production of such pigs requires a great deal of cost, time, and labor. Heterozygous ${\alpha}GT$ knockout pigs should be bred at least for two generations to ultimately obtain homozygote progenies. The present study was conducted to produce ${\alpha}GT$-deficient miniature pigs in much reduced time using mitotic recombination in neonatal ear skin fibroblasts. Methods: Miniature pig fibroblasts were transfected with ${\alpha}GT$ gene-targeting vector. Resulting gene-targeted fibroblasts were used for nuclear transfer (NT) to produce heterozygous ${\alpha}GT$ gene-targeted piglets. Fibroblasts isolated from ear skin biopsies of these piglets were cultured for 6 to 8 passages to induce loss of heterozygosity (LOH) and treated with biotin-conjugated IB4 that binds to galactose-${\alpha}$-1,3-galactose, an epitope produced by ${\alpha}GT$. Using magnetic activated cell sorting, cells with monoallelic disruption of ${\alpha}GT$ were removed. Remaining cells with LOH carrying biallelic disruption of ${\alpha}GT$ were used for the second round NT to produce homozygous ${\alpha}GT$ gene-targeted piglets. Results: Monoallelic mutation of ${\alpha}GT$ gene was confirmed by polymerase chain reaction in fibroblasts. Using these cells as nuclear donors, three heterozygous ${\alpha}GT$ gene-targeted piglets were produced by NT. Fibroblasts were collected from ear skin biopsies of these piglets, and homozygosity was induced by LOH. The second round NT using these fibroblasts resulted in production of three homozygous ${\alpha}GT$ knockout piglets. Conclusion: The present study demonstrates that the time required for the production of ${\alpha}GT$-deficient miniature pigs could be reduced significantly by postnatal skin biopsies and subsequent selection of mitotic recombinants. Such procedure may be beneficial for the production of homozygote knockout animals, especially in species, such as pigs, that require a substantial length of time for breeding.

• Reproductive and Developmental Biology
• /
• v.32 no.3
• /
• pp.153-158
• /
• 2008

Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

• Applied Microscopy
• /
• v.40 no.4
• /
• pp.193-200
• /
• 2010

This experiment was performed to evaluate the morphological responses of the duodenal epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of Bacillus Calmette-Guerin (BCG). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8\sim0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of $0.7{\mu}Ci$/g of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab, England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and duodenal tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab, England) in a dark room and dried and were placed in a light-tight box. The number of labeled epithelial cells in the duodenal mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. On the light microscopic study, duodenal mucosae had no severe morphological changes following the injection of BCG. In the tumor control and BCG treated groups, a number of small lymphocytes and eosinophile leucocytes are slightly increased as compared with those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 632.3 (${\pm}14.47$), 761.7 (${\pm}27.65$) and 505.0 (${\pm}17.09$) respectively. From the above results, BCG may suppress the DNA synthesis of the duodenal epithelial cells, but does not results severe structural defect on the duodenal mucosae. And it is suggested that BCG may greatly improve the anticancer therapy on certain kind of cancer.

• Journal of Animal Environmental Science
• /
• v.1 no.1
• /
• pp.9-19
• /
• 1995

This study was carried out to investigate the effect of sawdust bedding barn on manure handing, management labour and physiological aspect of dairy cows, and then to establish the criteria on the optimum utilization method of sawdust bedding barn. 46 tie stall barns and 49 sawdust bedding barns were surveyed to cmopare the milk productivity between two different barns, and also 5 tie stall barns, 15 sawdust bedding barns and 1 rice hull bedding barn were selected to study the utilization situation of sawdust barn in Kyung-Ki province area. The major results obtained were as follows; 1. The roof material of sawdust barn were consisted of 66% vinyl house, 23% PVC light and 11% slate and galvanum coated tin. Most of the floor structure was earth ground with the rate of the approximately 82%. 2. The average occupied area of sawdust bed per cow was 15.2 $m^2$, depth that 30 cm and the utilization period was 12 months. 3. Milk Yield was significantly higher at sawdust barn than at tie stall barn(P<0.01). Bacterial and somatic cell count in raw milk were less at the sawdust barn than in tie stall barn. However, there was not significance difference between two barns. 4. The labour hour needed to cow management in the sawdust barn was approximately 48% of that of tie stall barn. 5. The temperature and moisture content measured in sawdust bed were closely affected by seasonal ambient temperature. The skin and hair of cow were much cleaner at the PVC light roofed sawdust barn than any other roof materials. 6. The additives used for improving of fermentation did not show any effect on temperature and moisture content in sawdust bed. When the ambient temperature was $30.4^{\circ}C$, the surface temperature of measured 1 cm above the sawdust bed was $12.2^{\circ}C$ lower and the temperature of 100 cm above the sawdust bed was $2.4^{\circ}C lower under shading net facility than that of vinyl roofed one.7. The hoof length of miking cow was 7.95 cm in tie stall and 9.19 cm in sawdust barn with high significance (P<0.01). And disease occurence such as mastitis and foot-rot tended to decrease in the sawdust barn. 8. The number of total bacteria and coliform bacteria were less in the sawdust bed compared with earth ground resting area. And a parasite strongyloides papillosus was detected but without any infected cow. 9. The nitrate($NO_3N$) content in non-roofed earth ground resting area and earth ground under the sawdust bed was likely to pollute the ground water. 10. In economic point of view, rice hull bedding barn was the cheapest among different systems. And in the sawdust bedding barn PVC light + slate roofed barn was most desirable, and vinyl roofed one the least.

• Economic and Environmental Geology
• /
• v.51 no.4
• /
• pp.359-370
• /
• 2018

The batch and column experiments were performed to overcome the limitation of the neutralization process using the $scCO_2$-water-recycled aggregate, reducing its treatment time to 3 hour. The waste cement mortar and two kinds of recycled aggregate were used for the experiment. In the extraction batch experiment, three different types of waste mortar were reacted with water and $scCO_2$ for 1 ~ 24 hour and the pH of extracted solution from the treated waste mortar was measured to determine the minimum reaction time maintaining below 9.8 of pH. The continuous column experiment was also performed to identify the pH reduction effect of the neutralization process for the massive recycled aggregate, considering the non-equilibrium reaction in the field. Thirty five gram of waste mortar was mixed with 70 mL of distilled water in a high pressurized stainless steel cell at 100 bar and $50^{\circ}C$ for 1 ~ 24 hour as the neutralization process. The dried waste mortar was mixed with water at 150 rpm for 10 min. and the pH of water was measured for 15 days. The XRD and TG/DTA analyses for the waste mortar before and after the reaction were performed to identify the mineralogical change during the neutralization process. The acryl column (16 cm in diameter, 1 m in length) was packed with 3 hour treated (or untreated) recycled aggregate and 220 liter of distilled water was flushed down into the column. The pH and $Ca^{2+}$ concentration of the effluent from the column were measured at the certain time interval. The pH of extracted water from 3 hour treated waste mortar (10 ~ 13 mm in diameter) maintained below 9.8 (the legal limit). From XRD and TG/DTA analyses, the amount of portlandite in the waste mortar decreased after the neutralization process but the calcite was created as the secondary mineral. From the column experiment, the pH of the effluent from the column packed with 3 hour treated recycled aggregate kept below 9.8 regardless of their sizes, identifying that the recycled aggregate with 3 hour $scCO_2$ treatment can be reused in real construction sites.

• Korean Journal of Environmental Biology
• /
• v.23 no.3 s.59
• /
• pp.293-303
• /
• 2005

Japanese medaka, Oryzias latipes is widely distributed in the North East Asia including Korea, Japan and east China, and commonly used for freshwater toxicity tests and cytotoxicological studies worldwide. In this study, a series of experiments were conducted to identify the potential of the fish as a standard test species for saltwater toxicity evaluation such as marine receiving waters, ocean-dumped materials and sediment pore waters etc. Hatching, growth and mortality rates of the fish were estimated with the wide ranges of salinity from freshwater to seawater (35 psu). Direct exposure of the fertilized eggs in freshwater to the wide ranges of salinity (from 0 to 35 psu) without pre- acclimation to the saltwater revealed no significant differences in hatching rates by salinities (p =0.24). On the other hand, medaka larvae hatched in freshwater and exposed to saltwater directly showed high mortality at > 25 psu treatment groups (p < 0.0001). However, there was no significant difference in mortality of medaka larvae hatched in 13.8 and 14.2 psu at the wide ranges of salinities ($0\~35$ psu). Growth rates of medaka larvae hatched in the above two salinities showed no differences in body length either from 0 to 35 psu treatment groups (p =0.64 for 13.8 psu group and p=0.32 for 14.2 psu group). The number of gill chloride cell in medaka larvae sharply increased when the larvae were exposed to high salinity. Reference tests with zinc chloride revealed 96h $LC_{50}=8.84(7.19\~10.87)mg\;L^{-1}$ using 7~10 day old medaka larvae. These were comparable or better sensitivity in comparison with the other standard test species such as North American sheepshead minnow Cyprinodon variegatus. Based on the results of these experiments, hatching rates and larvalmortality of medaka must be good toxicity parameters for seawater bioassay and the species seems to be a good standard species for both the freshwater and seawater toxicity test.

• Applied Microscopy
• /
• v.32 no.1
• /
• pp.67-80
• /
• 2002

This study was carried out to investigate the effects of deer antler extract on the regeneration of peripheral nerves. Sprague-Dawley male rats weighing about 300 gm were fed deer antler extract for 1, 2, and 3 weeks per oral (1.5 ml/100 gm B.W.), respectively, once a day and transected both sides of sciatic nerve of each leg. After keeping for 6 hours, sciatic nerves taken from proximal part of transected region were treated with conventional transmission electron microscopical method and then observed with electron microscope. The results obtained were summarized as follows; 1. Sciatic nerves of normal control group were not showing any sprouts and electron dense axolemmal projections were frequently observed. 2. Sciatic nerves of saline treated groups were showing axonal sprouts at the nodes of Ranvier. The length of them was usually short, and numerous vesicles, vacuoles and organelles including neurofilament were contained. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 29 (29%) in 1 week treated group, 32 (32%) in 2 weeks treated group, and 30 (30%) in 3 weeks treated group, respectively. 3. Sciatic nerves of deer antler treated groups were showing axonal sprouts at the node of Ranvier as well. Although most of the sprouts were short, some sprouts of 2 weeks and 3 weeks treated groups were quite long. Sprouts usually contained numerous vesicles, vacuoles and cell organelles such as neurofilaments and mitochondria. The number of nodes of Ranvier containing sprouts from 100 longitudinal sectioned nerve fibers was 38 (38%) in 1 week treated group, 46 (46%) in 2 weeks treated group, and 48 (48%) in 3 weeks treated group respectively. The results described above explain pretreatment of deer antler extract improves the sprout formation of transected sciatic nerves, and then it suggests deer antler may be effective for the regeneration of peripheral nerves.