This study was designed to evaluate the possibility of increase through dairy female offspring's ratio by transfer of pre-selected transferrable blastocyst that was produced by pre-selected X-bearing semen with OPU derived oocytes. Elite dairy female cow is demanded strongly compared with male, the so called, farmer wants to produce only an elite female dairy offspring as a candidate female dairy cow for producing milk. In our study, we selected 2 elite dairy bull semen from National Agricultural Cooperative Federation to pre-select X-bearing semen and 5 elite dairy female cows as donor for collecting of OPU derived oocytes. OPU derived embryo production system was carried out an aspiration of immature oocytes from 5 donor cows 2 times per week, total 200 times for 2 to 7 months by an ultrasonographic guided follicular aspiration system and then produced in vitro-produced blastocysts by in vitro maturation, fertilization and culture. Dairy donor semen selected H-319, 320 bull in National Agricultural Cooperative federation was sorted X-bearing semen by flow-cytometer and frozen for using IVF with OPU derived oocytes. Donor cows were selected 5 elite dairy cows from Gyeongju Dairy Cow Community and then disease tests such as 4 kinds of disease before selecting was checked. Oocyte proportion of grade 1 to 3 from total collected oocytes was significantly lower in donor A and B than those in donor C, D and E (82.16 and 70.03% vs. 90.0, 91.78 and 93.57%), respectively (p<0.05). However, number of oocytes per session in donor A, C and E was significantly higher than those in donor B and D ($7.77{\pm}3.26$, $5.85{\pm}2.10$ and $7.03{\pm}2.14$ vs. $4.68{\pm}2.61$ and $5.21{\pm}1.97$ oocytes), but donor A was significantly higher than donor C (p<0.05). Development to blastocyst in donor B, C and E was significantly higher than those in donor A and D (31.0, 25.0 and 25.0% vs. 14.3 and 4.5%), but donor A was not different in donor C and E (p<0.05). Nine out of 10 blastocysts (90.0%) derived from OPU blastocysts were confirmed male embryos that was induced with Y-bearing semen to confirm sex ratio only. Total 96 blastocysts derived from female bearing semen were transferred into synchronized recipients and then confirmed 42 recipients (43.8%) pregnancy rate, 36 offspring (37.5%) and 91.7% female sex ratio (33 female vs. 3 male offspring). Taken together all data, elite dairy female offspring could be produced effectively by in vitro production system between pre-selected x-bearing semen and OPU derived oocytes that would be influential breeder in the breeding of dairy farm to increase effectively elite dairy offspring ratio as well as net income in the dairy farmer.
Purpose: The gluteal artery perforator flaps earned its popularity in buttock reconstruction due to the lower morbidity of the donor site and the flexibility in the design. Speedy and safe reconstruction is important for the success of buttock reconstruction. If a proper design is selected, satisfactory results can be obtained with more simple method of surgery. Methods: Between April 2005 and April 2006, buttock reconstruction by using gluteal artery perforator flaps were performed on sacral sores(6 cases), ischial sores(2 cases) and malignant melanoma on buttock(1 case). Various designs depending on the location and the size of the defect was made. In those designs, perforator was used as an axis for the minimal dissection of the vessel. Donor site from which sufficient amount of soft tissue can be transferred was selected, and also not causing high tension against the recipient site during the donor site closure. In addition, postoperative aesthetics, and the possibility of another design of a second operation which can be necessary in the future, was considered. Results: Patient follow up was for a mean period of 10.8 months. All flaps survived except for one that had undergone partial necrosis. Wound dehiscence was observed in one patient treated by secondary closure. Most patients presented with cosmetically and functionally satisfying results Conclusion: By designing the flap using the perforator as an axis, depending on the defect size and degree, reconstruction can be performed with only a small tension to the donor and the recipient site. And the minimal perforator dissection allowed easier and faster reconstruction. Selection of a proper design is the key procedure which greatly affects operation time and result success.
Cho, Bumrae;Kim, Su Jin;Lee, Eun-Jin;Ahn, Sun Mi;Lee, Jin Seok;Ji, Dal-young;Lee, Sang Hoon;Kang, Jung-Taek
Journal of Embryo Transfer
/
v.33
no.4
/
pp.245-254
/
2018
Pigs are considered as optimal donor animal for the successful xenotransplantation. To increase the possibility of clinical application, genetic modification to increase compatibility with human is an important and essential process. Genetic modification technique has been developed and improved to produce genetically modified pigs rapidly. CRISPR/Cas9 system is widely used in various fields including the production of transgenic animals and also can be enable multiple gene modifications. In this study, we developed new gene targeting vector and enrichment system for the rapid and efficient selection of genetically modified cells. We conducted co-transfection with two targeting vectors for simultaneous inactivation of two genes and enrichment of the genetically modified cells using MACS. After this efficient enrichment, genotypic analysis of each colony showed that colonies which have genetic modifications on both genes were confirmed with high efficiency. Somatic cell nuclear transfer was conducted with established donor cells and genetically modified pigs were successfully produced. Genotypic and phenotypic analysis of generated pigs showed identical genotypes with donor cells and no surface expression of ${\alpha}$-Gal and HD antigens. Furthermore, functional analysis using pooled human serum revealed dramatically reduction of human natural antibody (IgG and IgM) binding level and natural antibody-mediated cytotoxicity. In conclusion, the constructed vector and enrichment system using MACS used in this study is efficient and useful to generate genetically modified donor cells with multiple genetic alterations and lead to an efficient production of genetically modified pigs.
Nuclear transplantation techinque has been found to be the most potential and efficient method for producing a large number of genetically identical animals from a single embryo. The technical development of nuclear transplantation in mammals and its application to the production of cloned animals were reviewed. For the efficient and successful production of cloned embryos by nuclear transplantation, selection and micromanipulation of recipient eggs or embryos as capacious recipient cytoplasm, and benefitial preparation of multiple totipotent embryonic cells as donor nuclei, and also fusion technique are very critical. Recent works approaching to these critical points were introduced and discussed.
Son, Dae Gu;Kim, Hyun Ji;Kim, Jun Hyung;Han, Ki Hwan
Archives of Reconstructive Microsurgery
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v.13
no.1
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pp.43-50
/
2004
One of the major advantages of microsurgical reconstruction for defects of the hand is that these techniques allow for selection of the most ideal tissue to reconstruct a particular defect, thus optimizing the functional and aesthetic outcome. The dorsalis pedis free flap is an excellent reconstructive tool for various hand reconstructions. It has a reliable vasculature with vessels that are relative large on a long pedicle. It provides thin pliable tissue and be innervated by deep peroneal nerve. Coupled with its thinness and pliability, it is ideal for innervated cover of critically sensitive area, especially such as the hand. Thus it can be used as a cutaneotendinous flap, or an osteocutaneous flap. Otherwise, the major criticism with this flap is related to its uncertain vascularity and the donor defect. It is the purpose of this paper to outline our technique of flap elevation and donor site closure and to indicate our current use of this flap in hand reconstruction. We have treated 10 cases (6 burn scar contracture cases, 4 acute hand trauma cases) of hand reconstruction from Dec. 3, 1997 to Mar. 4, 2004 using dorsalis pedis free flap. The key points for sucess in terms of a viable flap and acceptable donor site are the preservation of the critical dorsalis pedis-first dorsal metatarsal vascular axis and the creation of a viable bed for grafting. In addition, we substituted preserved superficial fat skin graft for split thickness skin graft and wet environment was offered for good graft take. Preserved superficial fat skin is defined as composite graft containing epidermis, dermis and superficial fat layer. With sufficient care in flap elevation and donor site closure, a good graft take of preserved superficial fat skin under wet environment can be achieved with no functional disability and minimal cosmetic deformity in donor site. This flap has proved itself to be a best choice for hand reconstruction.
Liver transplantation is widely accepted as an effective therapeutic modality for a variety of irreversible acute and chronic liver diseases for which no satisfactory therapy is available. Following the first unsuccessful efforts at human liver transplantation in 1963, development of the procedure evolved at first slowly and steadily for 20 years and then rapidly over the past two decades. The growth of liver transplantation was facilitated by the conclusion of the national institutes of health consensus development conference in 1983 that liver transplantation is not an experimental procedure but an effective therapy that deserves broader application. The number of liver transplantations increased 2.4-fold(from 1.713 to 4.058) from 1988 to 1996. but the number of patients on the UNOS(united network of organ sharing) liver list increased 12.1-fold(from 616 to 7,467); as would be expected, the number of deaths of listed patients increased 4.9-fold(from 195 to 954), The current supply of donor livers is insufficient to meet this need, and organ donation has been stagnant or increased by only a few percent in recent years. These facts underscore the importance of the appropriate selection of candidates for liver transplantation and the development of operative procedures, such as living donor liver transplant, split liver transplant and auxiliary partial liver transplant.
Bari, Roberto Di;Coronelli, Roberto;Cicconetti, Andrea
Imaging Science in Dentistry
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v.43
no.3
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pp.135-143
/
2013
Purpose: This study was performed to obtain a quantitative evaluation of the cortical and cancellous bone graft harvestable from the mental and canine regions, and to evaluate the cortical vestibular thickness. Materials and Methods: This study collected cone-beam computed tomographic (CBCT) images of 100 Italian patients. The limits of the mental region were established: 5 mm in front of the medial margin of each mental foramen, 5 mm under the apex of each tooth present, and above the inferior mandibular cortex. Cortical and cancellous bone volumes were evaluated using SimPlant software (SimPlant 3-D Pro, Materialize, Leuven, Belgium) tools. In addition, the cortical vestibular thickness (minimal and maximal values) was evaluated in 3 cross-sections corresponding to the right canine tooth (3R), the median section (M), and the left canine tooth (3L). Results: The cortical volume was $0.71{\pm}0.23mL$ (0.27-1.96 mL) and the cancellous volume was $2.16{\pm}0.76mL$ (0.86-6.28 mL). The minimal cortical vestibular thickness was $1.54{\pm}0.41mm$ (0.61-3.25 mm), and the maximal cortical vestibular thickness was $3.14{\pm}0.75mm$ (1.01-5.83 mm). Conclusion: The use of the imaging software allowed a patient-specific assessment of mental and canine region bone availability. The proposed evaluation method might help the surgeon in the selection of the donor site by the comparison between bone availability in the donor site and the reconstructive exigency of the recipient site.
The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.
Agostini, Tommaso;Lo Russo, Giulia;Zhang, Yi Xin;Spinelli, Giuseppe;Lazzeri, Davide
Archives of Plastic Surgery
/
v.40
no.2
/
pp.91-96
/
2013
Background A thinned anterolateral thigh (ALT) flap is often harvested to achieve optimal skin resurfacing. Several techniques have been described to thin an ALT flap including an adipocutaneous flap, an adipofascial flap and delayed debulking. Methods By systematically reviewing all of the available literature in English and French, the present manuscript attempts to identify the common surgical indications, complications and donor site morbidity of the adipofascial variant of the ALT flap. The studies were identified by performing a systematic search on Medline, Ovid, EMBASE, the Cochrane Database of Systematic Reviews, Current Contents, PubMed, Google, and Google Scholar. Results The study selection process was adapted from the Preferred Reporting Items for Systematic Reviews and Meta-Analyses statement, and 15 articles were identified using the study inclusion criteria. These articles were then reviewed for author name(s), year of publication, flap dimensions and thickness following defatting, perforator type, type of transfer, complications, thinning technique, number of cases with a particular area of application and donor site morbidity. Conclusions The adipofascial variant of the ALT flap provides tissue to fill large defects and improve pliability. Its strong and safe blood supply permits adequate immediate or delayed debulking without vascular complications. The presence of the deep fascia makes it possible to prevent sagging by suspending and fixing the flap for functional reconstructive purposes (e.g., the intraoral cavity). Donor site morbidity is minimal, and thigh deformities can be reduced through immediate direct closure or liposuction and direct closure. A safe blood supply was confirmed by the rate of secondary flap debulking.
Ko, Kyung Jai;Kim, Young Hwa;Kim, Mi Hyeong;Jun, Kang Woong;Kwon, Kyung Hye;Kim, Hyung Sook;Kim, Sang Dong;Park, Sun Cheol;Kim, Ji Il;Yun, Sang Seob;Moon, In Sung;Hwang, Jeong Kye
Annals of Surgical Treatment and Research
/
v.95
no.5
/
pp.278-285
/
2018
Purpose: We investigated the clinical outcomes of deceased donor kidney transplantation (KT) using kidneys with terminal acute kidney injury (AKI). Methods: Between February 2000 and December 2013, we performed 202 deceased donor renal transplants from 159 brain dead donors. According to the expanded criteria donor (ECD) and AKI network criteria, we divided 202 recipients into 4 groups: Group I: Non-AKI & standard criteria donor (SCD) (n = 97); group II: Non-AKI & ECD (n = 15); group III: AKI & SCD (n = 52); and group IV: AKI & ECD (n = 38). Results: The incidence of delayed graft function (DFG) was significantly higher in patients with AKI than it was in the non-AKI group (P = 0.008). There were no significant differences among the 4 groups in graft survival (P = 0.074) or patient survival (P = 0.090). However, the long-term allograft survival rate was significantly lower in group IV than it was in other groups (P = 0.024). Conclusion: Allografts from deceased donors with terminal AKI had a higher incidence of DGF than did those from donors without AKI. However, there is no significant difference in graft and patient survival rates among the groups. So, the utilization of renal grafts from ECDs with terminal AKI is a feasible approach to address the critical organ shortage.
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