This study was performed during the four seasons for the production of transgenic pigs containing the human erythropoietin(hEPO) transgene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=42) were used fur the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality for zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9days after PG600 administration followed by superovulation with 1500IU pregnant mares serum gonadotropin (PMSG) and 500IU human chorionic gonadotrophin (hCG). Preparation of recombinant gene for microinjection is mice whey acidic protein promoter (mWAP) linked to human erythropoietin (hEPO) gene. After hormone treatment, 650 embryos were collected from 23 donors and 83.1% (540/650) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 543 DNA microinjected embryos fiom donors were transferred to 19 synchronized recipients, seven of them maintained pregnancy and delivered 47 piglets. One of the 47 offsprings were determined to have transgene by PCR analysis. The overall rate of transgenic production was 2.13% (tansgenic/offspring). This study provides the success and useful information regarding production of transgenic pig for bioreactor research.
This study was performed during the four seasons for the production of transgenic pigs containing the Cellulase Digest Gene. Purebred Landrace gilts and sows approximately 8∼15 months of age (n=126) were used for the collection of 1-cell zygotes for DNA microinjection and transfer. Retrospectively, estrus synchronization and superovulation schemes were evaluated to assess practicality fur zygote collection. Synchronization and superovulation procedures were used that cyclic gilts were synchronized with 20mg altrenogest (ALT) per day for 9 days after PG600 administration followed by superovulation with 1000 IU pregnant mares serum gonadotropin (PMSG) and 750IU human chorionic gonadotrophin (hCG). The cellulase digestion gene for microinjection is rat elasterase promoter (rEl) linked to CelD gene. After hormone treatment, 1,422 embryos were collected from 91 donors and 95.6% (1,359/1,422) embryos were in 1-cell stage which can be visualized the pronuclei for DNA microinjection. A total of 725 DNA microinjected embryos transferred into 35 recipients and produced 65 piglets from 13 litters. Pregnancy rate according to the number of transferred embryos to recipients was higher the group which received 21 to 24 embryos (50.0%) than other groups 20.0% in less and 33.3% in more. A tail tissue was collected from 65 piglets for biopsy. PCR screening was performed on each DNA sample using two separate sets of primers specific for the 5'- and 3'-flanking region of the rEl-CelD gene. Five of the 65 piglets (7.69%) were positive for the transgene. This study provide useful information regarding production of transgenic pig for bioreactor research.
This study was undertaken to investigate the effect of ovulation-synchronization(OVSYNCH) method on conception rate after insemination of dairy cattle. The proportion of conception of normal fertility cows(65%) was higher than that in infertility cows(50%) in dairy cows with OVSYNCH treatment. However, there was no significant difference between experimental groups. In another experiment, when the OVSYNCH method was adapted in multiparous dairy cows, the conception rate of experimental group of $30{\sim}35kg/day$ in milk yield was significantly higher than those in groups of $20{\sim}25kg/day$ in milk yield(p<0.05). In conception rates by OVSYNCH method in experimental groups with different parity, the group of 1st parity was significantly higher than in 3rd parity group(p<0.05). The results suggest that conception rate by OVSYNCH method was affected by various reproductive conditions in dairy cattle, and OVSYNCH method could be used to increase the conception rate for infertility cows with hormone disorder.
Kim, Chung-Hui;Han, Jin;Kim, Na-Ri;Park, Ju-Hee;Yang, Young-Churl;Kim, Eui-Yong
The Korean Journal of Physiology and Pharmacology
/
v.5
no.3
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pp.223-230
/
2001
Melatonin, a pineal gland hormone, is believed to act as an antioxidant via the stimulation of radical detoxifying enzymes and scavenging of free radicals. In this study, effects of in vitro and in vivo treatments of melatonin on the cisplatin-induced lipid peroxidation, LDH release and plasma creatinine were determined in rabbit renal cortical cells. The level of malondialdehyde (MDA) was assayed as an index of lipid peroxidation and the level of LDH release as an indicator of cellular damage. In in vitro studies, cisplatin increased the levels of MDA and LDH release in a concentration-and time-dependent manner. Melatonin inhibited the cisplatin-induced lipid peroxidation and LDH release in a concentration-dependent manner. The minimal effective concentration of melatonin that significantly reduced the $300\;{\mu}M$ cisplatin-induced lipid peroxidation and LDH release was 1 mM. In in vivo studies, the levels of lipid peroxidation and LDH release in renal cortical cells increased significantly 24 or 48 hours after a single injection of cisplatin (6 mg/kg). When the cisplatin-injected rabbits were pretreated with 10 mg/kg of melatonin, a significant reduction in both lipid peroxidation and LDH release was observed. The plasma creatinine level increased from $0.87{\pm}0.07$ mg/dl in control to $6.33{\pm}0.54$ mg/dl in cisplatin-injected rabbits (P<0.05). Melatonin partially prevented the increase in serum creatinine level $(1.98{\pm}0.11\;mg/dl)$ by cisplatin (P<0.05). In the proximal tubules from cisplatin-treated group, tubular cells had microvilli of variable heights. Necrotic debris was seen in tubular lumens. In most of cells, the mitochondria and lysosomes were increased in frequency. The endocytic vacuoles were not prominent and distribution of the brush border was irregular and shortened. These cisplatin-induced morphological changes were moderate in the melatonin-pretreated group. These results suggest that the toxicity of cisplatin is associated with the generation of reactive oxygen free radicals and that melatonin is a powerful antioxidant, which prevents some of the adverse effects of cisplatin.
Endocrine disrupting chemicals (EDCs) can alter hormone regulation that control reproductive system in animals. The effects of endosulfan, molinate, and alachlor that suspected to have examined disruption EDCs effect on a fish species of interest, Xiphophorus helleri (swordtail fish), were studied using vitellogenin (Vtg) and aromatase as diagnostic biomarkers. Induction of Vtg proteins was detected by RT-PCR in male fish treated with alachlor, and mixture of endosulfan and molinate in dose response manner. Also, induction of aromatase was detected by RT-PCR in male fish treated with alachlor, endosulfan, and mixture of endosulfan and molinate in sinlilar manner. In this study, swordtail fish exposed to endosulfan or molinate individually did not show any adverse effects. However, Vtg and aromatase expressions and apoptosis were detected in swordtail. fish exposed to the mixture of endosulfan and molinate. These results suggested that low concentrations of mixture of molinate and endosulfan individually do not affect swordtail fish, but may influence genital system, and induce apoptosis.
The concept of innate immunity in insects which refers to the first line of host defense constitutes the humoral and cellular components which are involved in recognition and actively participate in the elimination of the intruding foreign micro- or macro-organisms. Several recent studies suggest that juvenile hormone (JH) modulates the cellular immune reactions in response to pathogen. In this study, pyriproxyfen (a JH agonist as an insect growth regulator) was tested in its any inhibitory effect on the immune reactions of the beet armyworm, Spodoptera exigua. To this end, five different hemocyte morphotypes of final instar S. exigua were identified by phase contrast microscopy. Plasmatocytes and granular cells, which constitute about 90% of the total hemocyte count, were prominently distinguished based on their basophilic/acidophilic nature using Giemsa stain. The role of pyriproxyfen on the functional ability of hemocytes was analyzed using FITC-labeled Providencia vermicola for the phagocytic potential of the hemocytes. Both granular cells and plasmatocytes exhibited phagocytosis behavior. Pyriproxyfen significantly inhibited the phagocytosis of both cell types, proposing its novel action as an immunosuppressant.
It has been known that the proteome profiles in the period of growth and development of rice are changed by the growth conditions including temperature, soil, and fertilization. In this study, the proteome profiles of Odae polished white rice grown in Chulwon and Chilgog were compared on 2-dimensional(D) gels. The differentially expressed proteins were selected from the 112 identified total proteins and classified into functional groups. The most significantly differentially expressed proteins were stress responsive proteins; Ent-kaur-16-ene synthase, which is responsible for synthesizing a plant hormone gibberellin, was expressed in Chulwon rice and heat shock proteins were in Chilgog rice, respectively. Xylanase inhibitor protein, which inhibits the enzyme xylanase produced by pathogenic fungi and Bacilli, was expressed significantly high in Chilgog rice grown at high temperature. Differential expressions of transporter proteins were observed both in Chulwon and Chilgog rice. Regarding the facts that Chilgog rice contained relatively higher amount of proteins than Chulwon rice and Chulwon rice showed large number of proteins were differentially expressed, it can be concluded that different cultivation conditions could change the protein expression profiles in rice in various ways, including elevation of protein amount or differential expressions of specific proteins, etc. The results suggest that the characteristics of the profiles of the proteome in the polished white rice are definitely changed by the environmental factors including high temperature. The results can be utilized for the development of the proper cultivation conditions for the production of high quality rice with good palatability.
The sea bass, Lateoiabrax japonicus is a highly valued fish in aquaculture in South Korea. For establishment of seedling production of sea bass,1 japonicus, we examined change of gonadal development and plasma steroid levels of sea bass reared in indoor tank. Male matured unsimultaneously faster than females and spawning of females took place between the end of January and March. After the spawning period, and until the following January, all the females were in preyitello genesis and in some males, spermatogenetic activity restarted gradually. In October, under reducing photoperiod, cortical alveoli appeared in growing oocyte and the development of spermatogenesis greatly increased. Between October and february, vitellogenesis and spermatogenesis occurred respectively in female and male and gonadosomatic index increased from 4.31 to $24.07\%$ in female and upper 6o/o in male. Also, two sex hormones were analyzed during the course of a reproductive cycle in the sea bass: plasma levels of the gonadal steroid testosterone (T) and estradiol-l7$\beta$ (E_{2}). Variation of the plasma concentrations of T and E, appeared to depend on gonad stages. Plasma T and E, levels were high from November to January, suggesting that an sufficient gonadal stimulation by both hormones may undergoing a processes for the formation of sperm and oocyte.
Kang Duk Young;Chang Young Jin;Hur Jun Wook;Min Byung Hwa
Korean Journal of Fisheries and Aquatic Sciences
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v.35
no.6
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pp.551-556
/
2002
We have experimentally evaluated the content of thyroid hormones (THs), the growth and activity of larval rockfish (Sebastes schiegeli) born from parturient mother fish injected with 20 $\mu$g/g BW of 3,5,3-triiodo-L-thyronine ($T_{3}$). There was no difference of L-thyroxine ($T_{4}$) levels between controls (sham control and control) and $T_{3}$ groups in yolk-sac larvae just born from broodstock, while $T_{3}$level of $T_{3}$ group was significantly increased compared with controls. In addition, the both larval $T_{3}$ and $T_{4}$ levels in $T_{3}$ group were always higher than those of control during experimental period. Also, there were significant differences in the development of larvae from the two groups. The larval growth in total length and body weight according to the elapsed days after parturition showed the linear and curve equations, respectively, and the slopes of $T_{3}$ group were significantly higher than those of control. The survival rate of larvae in $T_{3}$ group was higher than that of control. Although the survival rate in $T_{3}$ group under the condition of starvation was significantly depressed compared with the control, larval swimming index in $T_{3}$ group was higher than control. Finally, these findings suggest that the exogenous $T_{3}$ could be transferred into hatched larvae in parturient rockfish by maternal injection, and subsequently the exogenous hormone could play some roles on physiological metabolism of larvae after parturition and may confer a distinct advantage to the fragile larvae during the early developmental stage.
Aleurone layer of Black rice (Oryza sativa L.) is enriched with anthocyanin that could increase bone density. This study was conducted to investigate the osteoporosis-preventing effects of the aleurone layer extract (BRE) on bone loss of ovariectomized (OVX) rats. OVX (or sham-operated) rats were assigned to three groups (n=8 per group): sham operated group (Sham); OVX control group (OVX); OVX-BRE group, OVX rats treated with BRE at 90 mg/kg B.W. The deionized water alone or deionized water with BRE was orally administrated to Sham, OVX or OVX-BRE groups, respectively for 12 weeks. High fat diet with 45 kcal% fat and water were fed to all rats ad libitum. Body weight was significantly decreased in the OVXBRE group compared to the OVX group (p<0.05). The bone mineral density and bone length of tibia were significantly higher in the OVX-BRE group compared to the OVX group and breaking force was significantly higher for the both tibia and femur bones. Serum estradiol concentration and calcium concentration of femur were higher in the OVX-BRE group than those of OVX group. However, serum alkaline phosphatase activity and parathyroid hormone concentration were decreased in the OVXBRE group compared to the OVX group. The results suggest that aleurone layer of Black rice is a potentially useful ingredient to protect against estrogen deficiency or menopause related osteoporosis.
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