Jungkeun Park

Cloning endangered gray wolves (Canis lupus) from somatic cells collected postmortem

The objective of the present study was to investigate whether nuclear transfer of postmortem wolf somatic cells into enucleated dog oocytes, is a feasible method to produce a cloned wolf. In vivo-matured oocytes (from domestic dogs) were enucleated and fused with somatic cells derived from culture of tissue obtained from a male gray wolf 6h after death. The reconstructed embryos were activated and transferred into the oviducts of naturally synchronous domestic bitches. Overall, 372 reconstructed embryos were transferred to 17 recipient dogs; four recipients (23.5%) were confirmed pregnant (ultrasonographically) 23-25 d after embryo transfer. One recipient spontaneously delivered two dead pups and three recipients delivered, by cesarean section, four cloned wolf pups, weighing 450, 190, 300, and 490g, respectively. The pup that weighed 190g died within 12h after birth. The six cloned wolf pups were genetically identical to the donor wolf, and their mitochondrial DNA originated from the oocyte donors. The three live wolf pups had a normal wolf karyotype (78, XY), and the amount of telomeric DNA, assessed by quantitative fluorescence in situ hybridization, was similar to, or lower than, that of the nuclear donor. In conclusion, the present study demonstrated the successful cloning of an endangered male gray wolf via interspecies transfer of somatic cells, isolated postmortem from a wolf, and transferred into enucleated dog oocytes. Therefore, somatic cell nuclear transfer has potential for preservation of canine species in extreme situations, including sudden death.

Improved efficiency of canine nucleus transfer using roscovitine-treated canine fibroblasts

The aim of this study was to investigate whether roscovitine (the cyclin-dependent kinase 2 inhibitor) effectively induces synchronization of the donor cell cycle at G0/G1 and to examine the effect of donor cell cycle synchronization protocols on canine somatic cell nucleus transfer. Canine fibroblasts were obtained from skin biopsy cultures taken from a 7-yr-old retriever. The donor cell cycle was synchronized either by culturing cells to reach confluency or by treating cells with 15 microg/mL roscovitine for 24h. Cell cycle stages and apoptosis were analyzed by flow cytometry. After synchronization of the donor cell cycle, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into 18 naturally estrus-synchronized surrogates. There was no significant difference in cell cycle synchronization and apoptosis rates between the confluent and roscovitine groups. After transfer of reconstructed embryos, pregnancy was detected in three of nine surrogates that received cloned embryos reconstructed with roscovitine-treated cells, whereas only one of nine surrogates was pregnant after transfer of cloned embryos reconstructed with confluent cells. One pregnant female from the confluent cell group delivered one live and one dead pup, but the live one died within 5 days after birth. Three pregnant females from the roscovitine-treated cell group delivered eight live pups and one dead pup, and one of eight live pups died within 6 days after birth. In conclusion, the current results demonstrated that reconstructing embryos with roscovitine-treated cells resulted in increased efficiency of canine somatic cell nucleus transfer.

Influence of oocyte donor and embryo recipient conditions on cloning efficiency in dogs

To determine factors that affect the efficiency of dog cloning by somatic cell nuclear transfer, the present study was performed to investigate 1) the effects of surgical history (non-operated/operated) and parity (nullipara/multipara) on the recovery of in vivo canine oocytes; 2) the effects of surgical history and parity of recipients on the pregnancy and delivery; and 3) the effects of synchronization state (AA, advanced asynchrony; SY, synchrony; RA, retarded asynchrony) between oocytes donor and recipient on the pregnancy and delivery. Oocyte recovery rate was significantly higher in non-operated dogs compared to operated dogs (93.8 vs. 89.6%, P < 0.05) and not different between nulliparous dogs and multiparous dogs. Delivery rate was also significantly higher in non-operated dogs compared to operated dogs (2.8 vs. 1.0%, P < 0.05) and in nulliparous dogs than multiparous dogs (3.0 vs. 1.7%, P < 0.05). Even though SY showed increased pregnancy and delivery rate (20.0% and 3.0%) compared to AA (15.0% and 2.0%) and RA (0.0% and 0.0%), there was no significant difference. In conclusion, we recommend non-operated dogs as experimental dogs and nulliparous dogs as recipient dogs to increase delivery rate after transfer of somatic cell nuclear transferred embryos, but further study is needed to find out appropriate synchrony status at the transfer.

Obstacle avoidance for small UAVs using monocular vision

Purpose – The purpose of this paper is to propose that the three‐dimensional information of obstacles should be identified to allow unmanned aerial vehicles (UAVs) to detect and avoid obstacles existing in their flight path.Design/methodology/approach – First, the approximate outline of obstacles was detected using multi‐scale‐oriented patches (MOPS). At the same time, the spatial coordinates of feature points that exist in the internal outline of the obstacles were calculated through the scale‐invariant feature transform (SIFT) algorithm. Finally, the results from MOPS and the results from the SIFT algorithm were merged to show the three‐dimensional information of the obstacles.Findings – As the method proposed in this paper reconstructs only the approximate outline of obstacles, a quick calculation can be done. Moreover, as the outline information is combined through SIFT feature points, detailed three‐dimensional information pertaining to the obstacles can be obtained.Practical implications – The propo...

Vision-Based Indoor Localization for Unmanned Aerial Vehicles

Small unmanned aerial vehicles are cost-effective and easy to operate, and especially suitable in dangerous indoor environments. However, because GPS is not available in an indoor environment, indoor localization is a crucial problem in developing small unmanned aerial vehicles (UAVs). This paper suggests vision-based indoor localization for UAVs in GPS-denied environments. Our approach is based on image matching by applying the scale invariant feature transform algorithm.

Birth of viable puppies derived from breeding cloned female dogs with a cloned male

Since the establishment of production of viable cloned dogs by somatic cell nucleus transfer, great concern has been given to the reproductive abilities of these animals (Canis familiaris). Therefore, we investigated reproductive activity of cloned dogs by (1) performing sperm analysis using computer-assisted sperm analysis and early embryonic development, (2) assessing reproductive cycling by measuring serum progesterone (P4) levels and performing vaginal cytology, and (3) breeding cloned dogs using artificial insemination. Results showed that most parameters of sperm motility in a cloned male dog were within the reference range, and in vivo-matured oocytes from a noncloned female were successfully fertilized by spermatozoa from a cloned male dog and develop normally to the 8-cell stage. Three cloned female dogs displayed normal patterns of P4 levels and morphologic changes of the vaginal epithelium. Two cloned female dogs became pregnant using semen from a cloned male dog and successfully delivered 10 puppies by natural labor. In conclusion, these data demonstrated that both cloned male and female dogs are fertile, and their puppies are currently alive and healthy with normal growth patterns.

Scheduler-Assisted Prefetching: Efficient Demand Paging for Embedded Systems

Embedded systems tend to use demand paging in order to provide more memory to applications in a cost-effective manner. However, demand paging drastically degrades the performance when the page fault rate is high. Prefetching has been known as a common remedy for page fault overhead. Although many prefetching mechanisms have been proposed, they are either effective only for specific page access patterns or too straight-forward to decrease a page fault rate to an acceptable level. We propose a scheduler-assisted prefetching mechanism which does not have such fundamental defects. As a proof of concept, our mechanism was completely implemented in Linux. We have also conducted a series of experiments to show its effectiveness. The experimental results showed a significant improvement: the number of the major page faults and the scheduling latency decreased by 30% and 51%, respectively.

Efficient User-Level I/O in the ARX Real-Time Operating System

User-level I/O gets increasingly important for embedded realtime applications, since it can allow programmers to write flexible and efficient device drivers for proprietary devices. To support user-level I/O for embedded systems, an operating system must provide a mechanism to deliver an external interrupt from an I/O device to a process in a predictable and efficient manner. In this paper, we propose an efficient user-level I/O scheme which is based on a newly designed user-level signal mechanism. This scheme which exploits the multithreading architecture of the kernel such as dynamic stack binding and scheduling event upcalls can overcome the problems of traditional signal implementations. We have fully implemented our scheme on the ARX real-time operating system that we already developed, and performed experiments to demonstrate its performance. The experiments clearly show that our user-level I/O scheme allows for a predictable delivery of external interrupts with a low overhead.

Lessons learned from cloning dogs.

The aim of this article is to review dog cloning research and to suggest its applications based on a discussion about the normality of cloned dogs. Somatic cell nuclear transfer was successfully used for production of viable cloned puppies despite limited understanding of in vitro dog embryo production. Cloned dogs have similar growth characteristics to those born from natural fertilization, with no evidence of serious adverse effects. The offspring of cloned dogs also have similar growth performance and health to those of naturally bred puppies. Therefore, cloning in domestic dogs can be applied as an assisted reproductive technique to conserve endangered species, to treat sterile canids or aged dogs, to improve reproductive performance of valuable individuals and to generate disease model animals.

Effects of Activin A on the In Vitro Development and mRNA Expression of Bovine Embryos Cultured in Chemically-Defined Two-Step Culture Medium

The purpose of the present study was to evaluate the effects of activin A on the developmental competence of in vitro fertilized (IVF) bovine embryos derived from a two-step defined culture system (C1/C2 medium) during the early or later stages of embryo development. To evaluate the effects of activin A on transcriptional levels, we analysed genes related to blastocyst hatching and implantation and to activin signalling pathway in IVF embryos. Cumulus-oocyte complexes were matured for 22 h and fertilized in vitro. Presumptive zygotes were cultured in the presence or absence of activin A during early (0-120 h, C1) or later (120-192 h, C2) stages. Although the developmental competence of embryos cultured with activin A in C1 medium was not significantly different from their corresponding controls, development to blastocysts (22.4% vs 34.7%; p < 0.05) and the blastocyst hatching rate (9.3% vs 22.4%; p < 0.05) in C2 medium supplemented with 100 ng/ml activin A were significantly higher than in the control group. To evaluate the effect of activin A on transcription, the relative expression levels of genes related to blastocyst hatching and implantation (Na/K-ATPase, E-cad and Glut-1) as well as activin signalling pathway (ActRII, ActRIIB and Smad2) were analysed. Compared to control medium, gene expression of Na/K-ATPase, E-cad, Glut-1, ActRII and ActRIIB was increased in medium supplemented with activin A. In conclusion, this study suggests that activin A, during the later stage of in vitro bovine embryo development, can enhance in vitro development of embryos by increasing hatching rates and affecting expression levels of genes related to hatching and implantation in defined culture medium.