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  • ISSN: 2379-061X
    Current Issue
    Volume 5, Issue 1
    Mini Review
    Chrenek P*
    Although, the first transgenic animal was produced more than 30 years ago, there are still many factors limiting efficiency of transgenesis. One of them is the low rate of transgene incorporation into the genome of microinjected eggs. This mini-review summarizes recent research methods based on pronuclear microinjection of rabbit fertilized eggs in order to reach higher efficiency of transgene integration and expression.
    Research Article
    Kharche SD*, Juhi Pathak, Anuj Kumar Singh Sikarwar, Chetna Gangwar, Priyadharsini R, Ravi Ranjan, Goel AK, Jindal SK, and Chauhan MS
    The aim of this study to observe in vivo development of a chimeric embryo (4n/2n) produced by tetraploid complementation. Cumulus oocyte complexes (COCs, 585) were collected and matured in maturation media. Out of which 524 matured oocyes were randomly divided into two groups. Group 1(n= 296) were activated with 5M Calcium Ionophore for 5-7 minutes followed by treatment with 2.0 mM DMAP for 4 hr in mCR2aa medium while Group 2 (n= 228) matured oocytes were used for in vitro fertilization. Furthermore, inner cell mass (ICM) from hatched blastocysts of parthenogenetic activated embryos were used to produce ES cell-like cells while 2 cell embryos obtained from IVF were used for their production of tetraploid embryos. Chimeric embryos were produced by microinjection of parthenogenetic ESCs in to in-vitro fertilized tetraploid caprine embryos. The percentage of 2 cell, 4 cell, 8-16 cell, morula, hatched blastocyst and cleavage rate of parthenogenetic goat embryos were 20.03 5.09%, 20.02 6.30%, 24.41 4.12%, 25.61 7.63%, 9.92 3.97% and 72.78 13.39%, respectively while the cleavage rate following IVF was 37.08 4.34%. Furthermore, 75.55 12.37% embryonic stem cell colonies were formed from hatched blastocysts. The IVF two cell embryo used for tetraploid embryo production, resulting in 75.09 7.86% fusion rate and 86.5 13.02% cleavage rate. A total of eight microinjected chimeric embryo were transfer in two naturally synchronized goat. The result indicated that caprine ES-like cells can be incorporated into IVF tetraploid embryos by microinjection so as to produce chimeric embryos. However, in vivo development of chimeric embryos could not achieve.
    Yong Zhang, Sheng Ya Guo, Xiao Yu Zhu, Juan Zhou, Wen han Liao, Gao Li Zheng, Ye Zuguang, and Chun Qi Li*
    Thrombosis is a prominent cause of many cardiovascular diseases, novel anti-thrombosis drugs are still in emergency. However, the development of effective and safe therapeutic agents for thrombotic diseases faces various challenges, especially on animal model. Here, we described the development and validation of a new thrombosis model induced by arachidonic acid (AA), a platelet-coagulation agonist, for drug screening and ef?cacy assessment, using larval zebrafish which were transparency in their early life period. Zebra?sh at 3 days post fertilization (dpf) were pre-treated with tested drug for 3 hours (h), followed by AA treatment at a concentration of 80M for 1.5 h. Tested drugs were administered into zebra?sh either by direct soaking or circulation microinjection. Antithrombotic efficacy was quantitatively evaluated based on an image analysis of the heart red blood cells (RBCs) stained with Odianisidine, and this technology has been patented previously. Thrombosis induced by AA in live zebrafish was visually confirmed under a dissecting stereomicroscope and quantified by the image assay. All the six human antithrombotic drugs (aspirin, clopidogrel, diltiazem hydrochloride injection, xueshuantong injection, salvianolate injection, astragalus injection) showed signi?cant anti-thrombotic effects (p < 0.05, p < 0.01 & p < 0.001) in this zebrafish thrombosis model. In this study, we introduced a new larval zebra?sh thrombosis model, which would be an attractive alternative for in vivo thrombosis studies and for rapid screening and efficacy assessment of antithrombotic drugs.
    Short Communication
    Jerome A. Werkmeister, Franca Casagranda, Glenn A. Edwards, Jacinta F. White, Veronica Glattauer, and John AM Ramshaw
    There is a significant unmet need for a biomaterial that is suitable for use as a small diameter vascular prosthesis for coronary by-pass surgery. The Omniflow Vascular Prosthesis? has proved clinically successful at larger diameters, for example for femoral artery replacement. The present study has examined whether type V collagen can provide an improved surface with poor thrombogenic properties that may assist in development of a coronary by-pass device. Biotinylated type V collagen was bound to biotinylated Omniflow Vascular Prosthesis? through use of a streptavidin bridge. The treated prosthesis showed a major reduction in the extent of platelet attachment, indicating that type V collagen coating should assist in the development of small diameter prosthesis.
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