This study showed the proper pigment extract in the Phe. Dyeing which is the basic dye of the natural plants dyeing,. Also it compared and examined the K/S the promotion of the color fastness and the effect of color difference change were attended by mordants and method of mordanting. 1. The most absorbance of Phellodendron. sol-ution is 330nm berberine is 430nm. 2. The best and proper temperature to extrat Phe,. was 80 and dyeing solution for 24 hours. 3. The pH effect was stable in the absorbances and color differences change. 4. Mordanted group increased the absorbances compared to non-mordanted sample. In case depth the most absorbance of Al. Sn. K. Cu and C. A solution were in 3% Fe and Cr were in 5%. In the color difference change they became dark reduced greenish and increased yellowish when their absorbances were in 3% 5. K./S was recoginzed by tainnin and fe, K/S was more increased pre-mordants than post-mordants. 6. Fe is the unchangable mordant in the color difference change and pre-mordants is more stable than post-mordants in the difference color. 7. Generally light C. F indicated more than grade 2 and light C. F, was much improved in post-mordanted by alumium potassium potass-ium dichromate. Washing C.F was improved in mordanted by tainnin and ferrous suulfate mordanted group was better than non-mordanted sample. Perspir-ation C,F was more stable in acid than alkali. Dry cleaning C.F indicated more than grade. 4.
Two 2 and 7 months-old holstin and one 3 months-old native Korean calf suffering from respiratory illness were submitted to the National Veterinary Research Institute for euthanasia and pathologic examination. At necropsy diffuse lobar pneumonia was present in apical cardiac and diaphragmatic lobes in all calves. Microscopically acute multifocal bronchiolitis and interstitial syncytial cell formation were frequently observed. in addition occlusion of bronchiolar lumen due to cellular proliferation and fibrosis accompanied by interlobular septal emphysema were also present. Immunohistochemically bovine respiratory syncytial virus antigen were positively identified in lung lesions including bronchial and bronchiolar epithelium alveolar macrophages and lymphocytes.
The ultimate goal of periodontal therapy is the regeneration of the periodontium that have been destroyed as a result of periodontal disease. This study were done in order to determine the healing status of periodontium under Polytetrafluoroethylene and millipore fillter combined with fibrin and the effect of the guided tissue regeneration procedures were performed as follows : 1) flap operation using PTFE membrane(control group) 2) flap operation using PTFE membrane which was fixed with fibrin(experimental group 1) 3) flap operation using millipore filter which was fixed with suture(experimental group II) 4) flap operation using millipore filter which was fixed with fibrin(experimental group II) After 1, 2, 4, 8, 12 weeks, dogs were sacrificed by perfusion technique and tissue block was excised including the tooth and prepared for light microscope with H-E & Masson’s trichrome staining. The result were as follows : In control and experimental group, there is no siginificant difference on epithelial cell down growth within 1st week, but more epithelial cell downgrowth in millipore or millipore combined with fibrin group. In this experiment, there were no significant difference in new cementum and alveolar bone formation whether PTFE membrane was fixed with suture or fibrin. In control and each experimental group, bone maturation appeared in 4 weeks, bone width increased bucco-lingually in control and experimental 1 group especially. Both control group and experimental group showed mild mew cementum formation on root surface and irregular arrangement of collagen fiber at 4 weeks, that showed obvious increased cementum formation at 8 weeks, and that was observed the functional arrangement of collagen fiber between new cementum and new alveolar bone at 12 weeks.
We investigated gonadal development and sex ratio of artificial seedlings of the oblong rockfish Sebastes oblongus, based on samplings for 370 days just after parturition. The primordial germ cells and genital ridge appeared separately under the mesentery in the yolk-sac stage larva (total length: 7.10-7.77 mm) just after parturition. The primordial germ cells and genital ridge integrated to form primordial gonad in 5-day-old larvae (7.12-9.68 mm), and then proliferation of somatic cell and germ cell occurred in the gonad, which was maintained undifferentiated until 45-days after parturition (18.6-20.4 mm). The ovarian differentiation began in the larva of 50-days old (dab) after parturition (dap) (20.0-24.5 mm). The somatic tissues elongated from the both opposite end-sites of undifferentiated gonad were consequently fused and formed a complete ovarian cavity at 60-days old dap (25.5-32.0 mm). In 80-days old dap (37.3-47.2 mm), meiosis of oogonia occurred to be chromatin nucleolus stage oocyte. The perinucleolus stage oocytes appeared at in 130-days old dap (68.0-86.0 mm), and previtellogenic stage oocytes appeared in 370-days old dap (101.0-116.0 mm). Only female was observed in the artificially produced oblong rockfish in the present study. This result revealed the effect of higher temperature on the sex determination of the oblong rockfish..
This study was conducted to investigate the effect of $Ca^{2+}$ concentration in fusion medium on the fusion, nuclear morphology and the development of bovine somatic cell nuclear transfer embryos. Bovine skin cells were transferred into an enucleated oocyte and fused with cytoplasm in the fusion medium containing with 0.05 to 1.0 mM Cacl$_2$. Nuclear transfer embryos were activated with a combination of A23187 and cycloheximide. Nuclear transfer embryos were fixed at 3 h after fusion or cultured for 7 ~8 days. Fusion rate was significantly (P<0.01) increased by increasing the $Ca^{2+}$ concentrations in the fusion medium from 0.05 mM (56.6%) to 0.5 mM (50.1%) and 1.0 mM (84.3%). More than 80% of reconstituted embryos underwent premature chromosome condensation (PCC) with 0.05, 0.1 mM CaCl$_2$, whereas 54.5% and 59.3% of embryos formed pronucleus (PN) directly without PCC in the 0.5 and 1.0 mM CaCl$_2$, groups. Blastocyst formation rates were significantly (P<0.05) different between 0.1 mM and 1.0 mM CaCl$_2$groups. From the present result, it is suggested that the elevated $Ca^{2+}$ concentrations in fusion medium can enhance the fusion and blastocyst formation rates of bovine nuclear transfer embryos.bryos.
Kim, Young-Sun;Kim, Tack-Soo;Ham, Suon-Kyu;Ban, Su-Won;Lee, Chang-Eun
Asian Journal of Turfgrass Science
/
v.23
no.1
/
pp.111-122
/
2009
Compound fertilizer contained slow release nitrogen was used at golf course management because of promoting turfgrass growth and decreasing fertilizing labor. But, it was hardly published about turf grass growth by its effect and form like isobutylidene diurea(IBDU) and methylene urea(MU) in golf course management of Korea. This study was conducted to investigate the effect of compound fertilizer contained slow release nitrogen such as IBDU and MU on creeping bentgrasss and the change in nitrogen in the root zone after application of fertilizers at AM turfgrass research institute with soil analysis, chlorophyll content index, leaf area index and dry weight during 5 months from July to december in 2007 year. Fertilizer treatments were designed as following; non-fertilizer(NF), straight fertilizer(CF), treatment 1(MU), treatment 2(IBDU 1), and treatment 3(IBDU 2). The every treatments were arranged a randomized complete block design with three replications. Results obtained were summarized as follows: As relative to time after N fertilizers application, ammonium and nitrate in soil were increased, but T-N was decrease. $NO_3$-N and T-N content of slow release fertilizer(SRF), the treatment MU, IBDU 1 and IBDU 2, in soil was higher than that of CF, and the change of ratio of available N and T-N by elapsed time after fertilizing SRFs and CF was higher about 5% in root zone of SF to l5days, but in root zone of SRF from 30days about $5{\sim}10%$. Compared with NF, the turf color index was increased 6.5%, 6.7%, 5.9% and 5.5%, respectively, the chlorophyll content 33.2%, 38.4%, 35.1% and 37.1%, the dry weight 76.2%, 77.7%, 69.5 and 72.3% and shoot number 28%, 52%, 56% and 51%. The chlorophyll content index of MD, IBDU 1 and IBDU 2 was increased $2.6{\sim}5.2%$ than that of CF, shoot number $18{\sim}22%$ but turf color index and dry weight was analogous to that of CF. These results showed that the SRF application was supplied nitrogen for a long time in root zone by increasing an available nitrogen, and so bring turf growth such as chlorophyll content and shoot number in creeping bentgrass.
In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.
This study was carried out to confirm whether the developmental capacity of bovine mature oocytes frozen ultra-rapidly using electron microscopic(EM) grids and EFS30 can be obtained, and whether the cryoprotectants and the freezing method used in this study effect detrimentally to the bovine oocytes by indirect immunocytochemistry. As freezing solution, we used EFS30 which consisted of 30% ethylene glycol, 0.5 M sucrose, 18% ficoll and 10% FBS added in D-PBS. The results obtained in this experiment were summarized as follows: When the effects of cryoprotectant and freezing procedure on the microtuble, micrfilament and chromatin morphology of oocytes were evaluated using indirect immunocytochemistry, the results of freezing as well as exposure group were not different with that of the control oocytes. When the fertilization abnormality after ultrarapid freezing of bovine mature oocytes was examined by Hoechst staining, the rates of total penetration(96.7, 9.0%), normal two pronuclei formation(74.6, 68.9%) and mean number of sperm / oocyte(1.50, 1.44) were not different between control and freezing group. In addition, when the developmental capacity of frozen-thawed of oocytes(85.5%) was survived, 74.5% of them were cleaved and 31.4% of cleaved embryos were developed to blastocyst. These data were similar to those of the control(76.0%, 34.6%) and exposure(74.5%, 33.0%) except survival rates. Also, when the total cell number of blastocysts produced from the each treatment at day after IVF was examined by hoechst staining, there were not different among groups. There results demonstrate that developmental capacity of frozen-thawed bovine mature oocytes can be successfully obtained by ultra-rapid freezing method using EM grid and EFS30 solution.
The dovelopment of the gonads, gametogenesis and the reproductive cycle of the topshell, Turbo cornutus Solander, which is one of valuable food animals fom Korean waters were studied by photomicroscophy. The materials were monthly collected from Bangeojin, Jeongjari and Dangweol, all these places being located in the south-eastern part of Korea, for one year from March 1979 to February 1980. Topshell is dioecious and oviparous. Gonad is situated on the surface of liver, which lies posteriorly. The surface of ovary and testis is covered with a fibrous membrane, membrane of connective and muscular fibers and then an outermost layer of simple-columnar epithelial cells which are composed of cuboidal and columnar mucous gland cells. Primordial germ cells develop on the germinal epithelium of ovarian and testicular lobuli which are originated from the fibrous membrane and extend toward hepatic gland. Undifferentiated mesenchymal tissue and pigment granular cells are abundantly distributed between the growing oocytes and spermatocytes in the early development stages. With the further development of the ovary and testis these tissue and cells gradually disappear. Then the undifferentiated mesenchymal tissue and pigment granular cells are considered to be related to the growing of the oocytes and spermatocytes. Early multiplicating oogonium is ca. $10\mu$ in diameter and nucleushaving a central nucleolus is ra. $8\mu$. As the oocytea grow to ca. $50-60\mu$ by the increase of cytoplasm, the oocytes become look like bunches of grapes which are attached to ovarian lobuli. Mature eggs are ca. $180-210\mu$ in diameter and it is surrounded by a gelatinous membrane of ca. $10\mu$ in thickness. After spawning, undischarged ripe eggs and spermatozoa remain in the ovary and testis respectively for some time. Then they finally degenerate, and proliferation of new oogonia and spermatogonia occur along the germinal epithelia of newly developed ovarian and testicular lobuli. Reprocuctive cycle of Turbo cornutus could be classified into five successive stages: multiplicative, growing, maturer spent and recovery stages. Spawning occurs from August to November with Peak spawning from early September to late October.
This study was to examine whether the in vitro friability, motility and intact acrosome of frozen-thawed bovine and human sperm can be improved by adding Pentoxifylline (PF) or Fertilization Promoting Peptide (FPP). Human semen was frozen ultra-rapidly using Test yolk-buffer (TYB) freezing medium. Additive (PF, FPP) effects in frozen-thawed bovine and human sperm were analyzed by microscopic count for sperm motility and coomassie brilliant blue staining method f3r sperm acrosome intact. The in vitro motility of frozen-thawed bovine sperm with 5 mM PF treatment group (50.0%) was significantly higher than that of control (34.0%) (P<0.05). In the frozen-thawed bovine sperm was examined, the intact acrosome rate of 50 nM FPP treatment (49.0%) was significantly higher than those of control (30.0%) and 25 nM FPP (38.0%) treatment groups (P<0.01). In human semen, when in vitro motility of sperm with PF addition prior to freezing was examined, the result of 5 mM treatment group (51.0%) was significantly higher than those of control and 2.5 mM treatment group (39.0, 40.0%) (P<0.01). In addition, 50 nM (75.5%) FPP adding in all treatment procedures for human semen freezing (before freezing, freezing and after thawing) was significant effect on maintenance of the sperm intact acrosome percentage (control: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P<0.01). Also, the intact acrosome rate of human sperm with FPP (65.0%) was significantly higher than that with PF (43.0%) (P<0.05), although sperm motility was slightly higher in PF treatment group. These results suggest that improved sperm motility and intact acrosome of frozen thawed bovine and human sperm can be obtained by addition of PF or FPP, and that the enhanced in vitro viability, motility and intact acrosome can be obtained by addition of FPP in all semen freezing procedures.
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