Byeong-Woo Kim

Investigation of de novo unique differentially expressed genes related to evolution in exercise response during domestication in Thoroughbred race horses.

Previous studies of horse RNA-seq were performed by mapping sequence reads to the reference genome during transcriptome analysis. However in this study, we focused on two main ideas. First, differentially expressed genes (DEGs) were identified by de novo–based analysis (DBA) in RNA-seq data from six Thoroughbreds before and after exercise, here-after referred to as “de novo unique differentially expressed genes” (DUDEG). Second, by integrating both conventional DEGs and genes identified as being selected for during domestication of Thoroughbred and Jeju pony from whole genome re-sequencing (WGS) data, we give a new concept to the definition of DEG. We identified 1,034 and 567 DUDEGs in skeletal muscle and blood, respectively. DUDEGs in skeletal muscle were significantly related to exercise-induced stress biological process gene ontology (BP-GO) terms: ‘immune system process’; ‘response to stimulus’; and, ‘death’ and a KEGG pathways: ‘JAK-STAT signaling pathway’; ‘MAPK signaling pathway’; ‘regulation of actin cytoskeleton’; and, ‘p53 signaling pathway’. In addition, we found TIMELESS, EIF4A3 and ZNF592 in blood and CHMP4C and FOXO3 in skeletal muscle, to be in common between DUDEGs and selected genes identified by evolutionary statistics such as FST and Cross Population Extended Haplotype Homozygosity (XP-EHH). Moreover, in Thoroughbreds, three out of five genes (CHMP4C, EIF4A3 and FOXO3) related to exercise response showed relatively low nucleotide diversity compared to the Jeju pony. DUDEGs are not only conceptually new DEGs that cannot be attained from reference-based analysis (RBA) but also supports previous RBA results related to exercise in Thoroughbred. In summary, three exercise related genes which were selected for during domestication in the evolutionary history of Thoroughbred were identified as conceptually new DEGs in this study.

Process design in flashless forging of rib/web-shaped plane-strain components by the finite element method

Abstract In this work preforming operations in the forging of rib/web-shaped plane-strain components are designed by the rigid-plastic finite element method to obtain flashless products, the height-to-width ratios of rib the geometry used for the analysis and design being 1.0, 2.0 and 3.0. The two design criteria of flashless geometrical filling and an even distribution of effective strain in the final products are investigated in controlling the preform configuration and establishing a systematic procedure of process design. One preforming operation is designed for flashless forging with a sound distribution of effective strains at ratios of 1.0 and 2.0. The case of a ratio of 3.0 also needs one preform to satisfy flashless geometrical filling, but has high effective strain, a further preforming operation needing to be added to avoid the high value of effective strain: thus the case of a ratio of 3.0 is designed as two preforming operations. The resulting preform configurations in plane-strain forging are compared with those of axisymmetric forging of rib/web components. It is noted that the flashless forging of plane-strain rib/web components requires a unique preforming operation to make the metal flow slower at the central part.

Effects of Palm Kernel Expellers on Growth Performance, Nutrient Digestibility, and Blood Profiles of Weaned Pigs

This experiment was conducted to investigate the effects of palm kernel expellers on growth performance, nutrient digestibility, and blood profiles of weaned pigs. A total of 88 weaned pigs (6.94±0.76 kg body weight [BW]; 28 d old) were randomly allotted to 2 dietary treatments (4 pigs/pen; 11 replicates/treatment) in a randomized complete block design (sex as a block). The dietary treatments were a typical nursery diet based on corn and soybean meal (CON) and CON added with 20% of palm kernel expellers (PKE). Pigs were fed for 6 wk using a 3-phase feeding program with declining diet complexity and with phases of 1, 2, and 3 wk, respectively. Blood was collected from randomly selected 2 pigs in each pen before weaning and on d 7 after weaning. Pigs were fed respective dietary treatments containing 0.2% chromic oxide from d 29 to 35 after weaning. Fecal samples were collected from randomly selected 2 pigs in each pen daily for the last 3 days after the 4-d adjustment period. Measurements were growth performances, digestibility of dry matter, nitrogen and energy, white and red blood cell counts, packed cell volume, and incidence of diarrhea. The PKE increased average daily gain (ADG) (246 vs 215 g/d; p = 0.06) and average daily feed intake (ADFI) (470 vs 343 g/d; p<0.05) and decreased gain-to-feed ratio (G:F) (0.522 vs 0.628 g/g; p<0.05) during phase 2 compared with CON, but did not affect growth performance during phase 1 and 3. During overall experimental period, PKE increased ADG (383 vs 362 g/d; p = 0.05) and ADFI (549 vs 496 g/d; p<0.05) compared with CON, but did not affect G:F. However, no differences were found on digestibility of dry matter, nitrogen, and energy between CON and PKE. The PKE reduced frequency of diarrhea (15% vs 25%; p = 0.08) for the first 2 wk after weaning compared with CON. Similarly, PKE decreased white blood cells (8.19 vs 9.56×103/μL; p = 0.07), red blood cells (2.92 vs 3.25×106/μL; p = 0.09), and packed cell volume (11.1% vs 12.6%; p = 0.06) on d 7 after weaning compared with CON. In conclusion, addition of 20% palm kernel expellers to nursery diet based on corn and soybean meal had no negative effects on growth performance, nutrient digestibility, and blood profiles of weaned pigs.

A Study on Genetic Parameters of Carcass Weight and Body Type Measurements in Hanwoo Steer

This study was conducted to estimate heritabilities and genetic correlations among carcass weight and body type measurements (Wither height, Rump height, Body length, Chest depth, Chest width, Rump length, Rump width, Thurl width, Hipbone width, Chest girth) obtained from 1,587 records of Hanwoo steers in National Hanwoo Contest. Based on EM-REML algorithm, this study estimated genetic parameters of the carcass weight and body type measurements that greatly affect Hanwoos carcass grade and body type. The means and standard deviations of each trait were 377.30±45.59 kg , 136.14±4.51 cm , 137.79±4.62 cm , 157.77±6.97 cm , 77.55±3.62 cm, 51.72±4.30 cm , 55.04±3.19 cm , 50.21±2.96 cm , 48.29±2.94 cm , 29.12±2.04 cm , and 217.31±11.36 cm for Carcass weight, Wither height, Rump height, Body length, Chest depth, Chest width, Rump length, Rump width, Thurl width, Hipbone width and Chest girth, respectively. As a result of analysis on the effects by the birth year, the Carcass weight, Wither height, Rump height, Body length, Chest depth, Chest width, Rump length, Rump width, Thurl width, and Chest girth came out highest as 413.4±37.5 kg , 138.4±4.1 cm , 139.9±4.3 cm , 162.1±5.7 cm , 80.8±2.5 cm , 55.3±3.7 cm , 58.0±3.3 cm , 52.9±2.4 cm , 50.9±2.8 cm and 225.9±8.3 cm , respectively in 2005. As a result of analysis on the effects by the birth season, the carcass weight and body type measurements came out highest, respectively in spring. As a result of the analysis on the effects by the birth location, all of the traits came out highest, respectively in Gangwon. In the majority of carcass weights and body type measurements, statistically significant differences were observed depending on the location of birth. The heritabilities of the carcass weight, chest width and chest girth were estimated as 0.242, 0.221 and 0.264 respectively. These heritabilities of such traits were higher than those of other traits (withers height, rump height, body length etc.). Genetic correlation coefficients were estimated to be 0.944 between wither height and rump height, 0.822 between carcass weight and chest width, 0.769 between carcass weight and chest girth. From the results above, if the carcass weight should be improved by body type measurements of Hanwoo steer, it seemed that such factors as, chest width, chest girth, body length and withers height, which have relatively higher genetic correlations with carcass weight, should be considered.

The effects of artificial activation timing on the development of SCNT-derived embryos and newborn piglets

This study investigated the effects of two different activation regimens on the developmental potential of somatic cell nuclear transfer (SCNT) embryos and postnatal survivability of the cloned piglets. In vitro matured oocytes were enucleated and reconstructed with porcine fetal fibroblasts. On the basis of the activation regimen used, the reconstructed porcine embryos were allocated into two groups: Group 1-simultaneous electrical pulses and activation group (SFA group); and Group 2-electrical fusion without calcium followed by electrical pulses with calcium after colcemid and cytochalasin B treatment for 5h (DA group). Embryonic development in both SFA and DA groups was determined at day 6 of culture in NSCU-23 medium. To investigate the post-implantation development after the two activation methods, embryos were cultured for 1 day and then transferred into the oviducts of estrus-synchronized recipients. DA group had significantly (p<0.05) higher cleavage rates than SFA group. However, the developmental rate to the blastocyst stage and the mean cell number of blastocysts did not differ (p>0.05) between SFA and DA groups. Moreover, the pregnancy rate of SFA group was not significantly different compared to DA group. A total of 20 cloned piglets (SFA group-8 live piglets, DA group-11 live piglets and one stillborn) were obtained in the present study. The birth weight of the cloned piglets (live births) did not differ (p>0.05) between the two groups. Furthermore, no difference was observed in the postnatal survival rates of the cloned piglets obtained using two different activation regimens. These results suggest that the timing of artificial activation and additional chemical treatments do not affect the developmental rate of porcine SCNT embryos. Remarkably, the pregnancy rate and postnatal survivability of the cloned piglets did not vary between SFA and DA groups.

The Effect of Body Measurements Type on Carcass Traits in Hanwoo

This study was carried out to investigate the correlation of body measurement type and carcass traits. This study used body measurement type and carcass traits on 1,312 heads of Hanwoo steers tested(24 months) which was from 1997 to 2002 at Hanwoo Improvement Main Center National Agricultural Cooperatives Federation. Body measurement type used the age of 18 months body measurement type, and carcass traits used the results of decision grade of Animal Products Grading Service. Observing the phenotypic correlation between carcass traits and body measurement type from the result, the weight, withers height, and chest width at age of 18 months and carcass weight showed readings of 0.690, 0.483, and 0.506, respectively. Also, eye muscle area and age of 18 months weight and chest width read 0.356 and 0.279, respectively. According to results, that improvements in weight, withers height, and chest width are likely to enlarge the eye muscle area and meat production level. Not only quality improvement but also quantity augmentation affect income and sales of the farmers. As can be seen in this study, improvements of weight, withers height and chest width should always be put to much effort while other body measurement type should be considered continuously too. In order to attain clearer results further studies should be done on body measurement type, which will contribute to the improvements of body measurement type in Hanwoo.

Influences of somatic donor cell sex on in vitro and in vivo embryo development following somatic cell nuclear transfer in pigs.

Objective The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture (143.8±10.5 to 159.2±14.8) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups (31.4±8.3 to 33.4±11.1). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05) between sexes. Conclusion The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.

Proteomic identification of abnormally expressed proteins in early-stage placenta derived from cloned cat embryos

It is unknown whether gene expression in cloned placenta during pre- and postimplantation is associated with early pregnancy failure in the cat. In this study, protein expression patterns were examined in early-stage (21-day-old) domestic cat placentas of fetuses derived from AI (CP; N = 4) and cloned embryo transfer (CEP; N = 2). Differentially expressed proteins were analyzed by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight (TOF) mass spectrometry (MS). A total of 21 proteins were aberrantly expressed (P < 0.05) by >1.5-fold in CEP compared with CP. Compared with CP, 12 proteins were upregulated in CEP (peptidyl-prolyl cis-trans isomerase A, annexin A2, protein DJ-1, adenylate kinase isoenzyme 1, protein disulfide-isomerase A3, actin cytoplasmic 1, serum albumin, protein disulfide-isomerase A6, and triosephosphate isomerase), and nine proteins were downregulated (triosephosphate isomerase; heterogeneous nuclear ribonucleoprotein H; tropomyosin alpha-4; triosephosphate isomerase 1; 60 kDa heat shock protein, mitochondrial; serum albumin; calumenin; keratin type 1; and prohibitin). The identities of the differentially expressed proteins were validated by peptide mass fingerprinting using matrix-assisted laser desorption/ionization-TOF/TOF MS/MS. The abnormally expressed proteins identified in this study might be associated with impaired development and dysfunction of CEP during early pregnancy. Abnormal protein expression might also induce fetal loss and contribute to failure to maintain pregnancy to term.

Molecular cloning, purification, expression, and characterization of β-1, 4-endoglucanase gene ( Cel5A ) from Eubacterium cellulosolvens sp. isolated from Holstein steers’ rumen

Objective This study was conducted to isolate the cellulolytic microorganism from the rumen of Holstein steers and characterize endoglucanase gene (Cel5A) from the isolated microorganism. Methods To isolate anaerobic microbes having endoglucanase, rumen fluid was obtained from Holstein steers fed roughage diet. The isolated anaerobic bacteria had 98% similarity with Eubacterium cellulosolvens (E. cellulosolvens) Ce2 (Accession number: AB163733). The Cel5A from isolated E. cellulolsovens sp. was cloned using the published genome sequence and expressed through the Escherichia coli BL21. Results The maximum activity of recombinant Cel5A (rCel5A) was observed at 50°C and pH 4.0. The enzyme was constant at the temperature range of 20°C to 40°C but also, at the pH range of 3 to 9. The metal ions including Ca2+, K+, Ni2+, Mg2+, and Fe2+ increased the endoglucanase activity but the addition of Mn2+, Cu2+, and Zn2+ decreased. The Km and Vmax value of rCel5A were 14.05 mg/mL and 45.66 μmol/min/mg. Turnover number, Kcat and catalytic efficiency, Kcat/Km values of rCel5A was 96.69 (s−1) and 6.88 (mL/mg/s), respectively. Conclusion Our results indicated that rCel5A of E. cellulosolvens isolated from Holstein steers had a broad pH range with high stability under various conditions, which might be one of the beneficial characteristics of this enzyme for possible industrial application.

Micellar core-shell-type acrylic-polyurethane hybrid materials with self-polishing property

Abstract Marine fouling can be a serious problem in the shipping industry, since it increases the surface roughness of the hull and hence its frictional resistance to its movement through water. Antifouling paint can be defined as preventing the attachment of marine organisms onto surfaces. However, the most commonly used antifouling coating which is the tributyltin-based self-polishing copolymer causes the severe pollution of marine environment. Ammonium salt-based paints include tertiary amines as biocides which have effective biocidal and biodegradable properties without accumulation in the sea environment. However, ammonium salt-based coatings were too sensitive to seawater and became swollen. In this study, polyurethane-acrylic copolymers were synthesized by radical polymerization. These hybrid materials were found to form core–shell structures in aqueous media. Synthesis and properties of copolymers were investigated by Fourier transform-infrared spectrometer, proton-nuclear magnetic resonance, transmission electron microscopy, and dynamic light scattering. The polishing rate of self-polishing copolymer was determined from the reduction in dry film thickness after artificial seawater immersion under a dynamic condition.