Recently, we published a microinjection method for generating transgenic cattle using the DNA transposon system and their analysis by next-generation sequencing (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In that study, we generated transgenic cattle using two different types of DNA transposon system, sleeping beauty (SB) and piggybac (PB), carrying Yellow fluorescent protein with SB (SB-YFP, female) and green fluorescent protein with PB (PB-GFP, male) under the control of the ubiquitous CAG promoter, respectively. The female and male founder cattle have been grown up to date (the female age: 40 months old, the male age: 33 months old) without any health issues. In genomic instability and blood analysis, there was no significant differences between wild type and founder cattle. In the present study, we confirmed germ-line transmission of the transposon-mediated transgene integrations and ubiquitous and persistent expression of transgene in second generation of offspring (F1). The F1 was born without any assistance and expressed GFP in the eyes without UV light. The ubiquitous expression of GFP was detected in skin fibroblast from the ear tissue and confirmed by genomic DNA PCR, which suggest that the transgene from the PB-GFP was successfully transmitted. Unfortunately, no transgene from SB-YFP were identified. To confirm the transgene integration site, the genomic DNA from blood was extracted and performed next-generation sequencing (NGS). The GFP gene was integrated in chromosome 4 (two copies), and 6. As results, a total of two copies of paternal transgene transmitted into the F1. All the integrated position was not related with coding region and there was no significant difference in genomic variants between transgenic and non-transgenic cattle. To our knowledge, this is the first report of germ-line transmission through non-viral transgenic founder cattle. Those transgenic cattle will be valuable resource to many fields of biomedical research and agricultural science.

CRISPR/Cas9-induced knock-out/-in can be occurred at specific locus in the genome by non-homologous end joining (NHEJ) or homology directed repair (HDR). Here, we demonstrate the targeted insertion into the specific loci of embryo fertilized by semen from transgenic cattle via CRISPR/Cas9 system. Recently, we published on the efficient generation of transgenic cattle using the DNA transposon system (Yum et al. Sci Rep. 2016 Jun 21;6:27185). In the study, eight transgenic cattle were born following transposon-mediated gene delivery system (Sleeping Beauty and Piggybac transposon system) via microinjection. In the analysis of their genome stability using next-generation sequencing, there was no significant difference in the number of genetic variants between transgenic and non-transgenic cattle. All the transgenic cattle have grown up to date (the oldest age: 33 months old, the youngest age: 15 months old) without any health issue. One of transgenic male cattle expressing GFP reached puberty and semen was collected. Over 200 frozen semen straws were produced and some were used for in vitro fertilization (IVF). On seven days after IVF, expression of GFP was observed at blastocyst stage and was seen in 80% of the embryos. Another application is to edit the GFP locus of the transgenic cattle because long-term and ubiquitous expression of transgene didn’t affect their health. In one cell stage embryos produced using GFP frozen-thawed semen, microinjection of sgRNA for GFP, Cas9, together with donor DNA that included RFP and homology arms to link the double-strand break of sgRNA target site into fertilized eggs resulted in expression of RFP. This indicated that the GFP locus of transgenic cattle shows potential candidates for stable insertion of the functional transgene. Knock-out/-in for editing GFP locus using CRISPR-Cas9 might be a valuable approach for the next generation of transgenic models by microinjection. In conclusion, we demonstrated P-112 that transgenic cattle via transposon system are healthy to date and germ-line competence was confirmed. The GFP locus will be used as the potential target site for future gene engineering via genome-editing technology. Finally, all those animals could be a valuable agricultural and veterinary science resource for studying the effects of gene manipulation on biomedical research and medicine. This work was supported by BK21 PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU 550-20160004).

This study was purposed to provide basic information on the correct application of a wheelchair's backrest angle by investigating the change in cardiopulmonary function according to backrest angle during propulsion. This study examined the effects of the wheelchair's backrest angle on the cardiopulmonary function by varying the angle to 0°, 10° and 20° and with a propulsion velocity of 60 m/min. The experimental parameters were respiration rate, oxygen consumption rate and oxygen consumption rate/kg which were measured by a portable wireless oxygen consumption meter (COSMED, K4b²). The results of the study were as follows: 1) There were no statistically significant differences in respiration rates due to changes in the wheelchair backrest angle (p>.05). 2) There were statistically significant differences in oxygen consumption rates due to changes in the wheelchair backrest angle (p<.05). 3) There were also statistically significant differences in the oxygen consumption rate/kg due to changes in the wheelchair backrest angle (p<.05). In conclusion, changes in the backrest angle of wheelchairs during propulsion influences oxygen consumption rates and heart rates, while respiration rates are not affected. Therefore, a training program for good seating and posture needs to be provided, and the wheelchair seating system should be equipped with the unadjustable-angle wheelchair to reduce the functional load on the cardiopulmonary system.

In the sea various methods have been conducted to capture wave energy which include the use of pendulums, pneumatic devices, etc. Floating devices, such as a cavity resonance device take advantages of both the water motion and the wave induced motions of the floating body itself. The wave energy converter is known commercially as the WAGB(Wave Activated Generator Buoy) and is used in some commercially available buoys to power navigation aids such as lights and horns. This wave energy converter consists of a circular flotation body which contains a vertical water column that has free communication with the sea. A theoretical analysis of this power generated by a pneumatic type wave energy converter is performed and the results obtained from the analysis are used for a real wave energy converter buoy. This paper is shown to have an optimum value for which maximum power is obtained at a given resonant wave period Also, the length of the internal water column corresponds to that of the water mass in the water column. If designed properly, wave energy converter can take advantage not only of the cavity resonance, but also qf the heaving motion of the buoy. Finally, simulation is performed with a LabVIEW program and the simulation results are applied to a wave energy simulator for modifying design data for a wave energy converter.

We described and illustrated a rare species in Korea, Hypoxis aurea Lour. (Hypoxidaceae) which was rediscovered about 70 years after its first collection from Jeju island in Korea. The members of the family Hypoxidaceae R. Br. are distinguished from the plants of Amaryllidaceae J. St-Hill. by having grass-like leaves, an invisible stem which is modified into a corm or a rhizome, trimerous, and radially symmetric flowers with an inferior ovary developing into a capsule on scapes. Hypoxis aurea Lour. is readily distinguishable from Curculigo orchinoides Gsertn. in Japan by beakless ovary and capsular fruit. The number of somatic chromosome is 2n=54.