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Biomedical subjects

W S Hwang

Publications and source records attributed to W S Hwang.

At least 19 recordsLinked to original sources
PubMed2006

Effects of combined expression of human complement regulatory proteins and H-transferase on the inhibition of complement-mediated cytolysis in porcine embryonic fibroblasts.

The expression of human complement regulatory proteins (CRP) and H-transferase (HT) in porcine cells is one of the strategies for suppression of hyperacute rejection (HAR) of xenotransplants in human recipients. In this study, we investigated the inhibitory effect of combined expression of human complement regulators and HT on human serum-mediated cytolysis in porcine embryonic fibroblasts. For the combinated expression of human CRPs in transformed pig cells, cDNAs of human DAF, MCP, and CD59 were cloned into the same insertional plasmid under the control of pCMV IE and LTR. The double combination of CRPs, hDAF-hMCP, and hMCP-hCD59 survived over 50% in the presence of 50% human serum, compared to the control. Moreover, the cell viability was increased more than 65% and 80% in the combination of human DAF-CD59 and DAF-MCP-CD59, respectively. In addition, the combination of HT gene to hDAF-hCD59 vector increased the viability close to 80%, similar to the triple combination of CRPs. These observations suggest that the combined expression of human CRPs and HT in the same insertional vector may be more effective in protecting porcine cells from human complement-mediated cytolysis.

Animals↗
PubMed2006

The effects of Goami No. 2 rice, a natural fiber-rich rice, on body weight and lipid metabolism.

OBJECTIVE: Increased intake of dietary fiber reduces the risk of obesity and type 2 diabetes. We assessed the effects of a fiber-rich diet on body weight, adipokine concentrations, and the metabolism of glucose and lipids in non-obese and obese subjects in Korea, where rice is the main source of dietary carbohydrates. RESEARCH METHODS AND PROCEDURES: Eleven healthy, non-obese and 10 obese subjects completed two 4-week phases of individual isoenergetic food intake. During the control diet phase, subjects consumed standard rice; during the modified diet phase, subjects consumed equal proportions of fiber-rich Goami No. 2 rice and standard rice. We used a randomized, controlled, crossover study design with a washout period of 6 weeks between the two phases. RESULTS: After the modified diet phase, body weight was significantly lower in both the non-obese and obese subjects (non-obese, 57.0 +/- 2.9 vs. 56.1 +/- 2.8 kg, p = 0.001; obese, 67.7 +/- 2.1 vs. 65.7 +/- 2.0 kg, p < 0.001 for before vs. after). The BMI was significantly lower in obese subjects (26.9 +/- 0.5 vs. 26.0 +/- 0.6 kg/m2, p < 0.001). The modified diet was associated with lower serum triacylglycerol (p < 0.01), total cholesterol (p < 0.01), low-density lipoprotein cholesterol (p < 0.05), and C-peptide (p < 0.05) concentrations in the obese subjects. DISCUSSION: These results indicate that fiber-rich Goami No. 2 rice has beneficial effects and may be therapeutically useful for obese subjects.

Adiponectin↗
PubMed2005

High oxygen tension during in vitro oocyte maturation improves in vitro development of porcine oocytes after fertilization.

This study was conducted to evaluate the effect of oxygen tension during IVM and/or IVC on developmental competence of porcine follicular oocytes. Prospective, randomized experiments were designed, and oocytes were matured, inseminated and cultured in vitro in the designated condition. In experiment 1, either high (20%) or low (7%) oxygen tension was used for IVM. The high oxygen significantly improved blastocyst formation (23% versus 13%; P<0.01) after IVF than the low oxygen. Such treatment, however, did not significantly (P>0.05) improve the rates of nuclear maturation (89% in each treatment), sperm penetration (62-72%), monospermic fertilization (56-67%), pronuclear formation (90-96%), cleavage (49-53%) and blastocyst cell number (31-32 cells). In experiment 2, the combined effect of oxygen tension during IVM and IVC of embryos was evaluated by a 2 x 2 factorial arrangement. Again, the high oxygen tension during IVM supported blastocyst formation more efficiently (P<0.01) than the low oxygen, and this was independent of oxygen tension during IVC (26-28% versus 15-16%). In oocytes matured under the high oxygen, a tendency to increase blastomere number (P=0.0630) was found, when the low oxygen was used for IVC after insemination (39-45 cells/blastocyst). In conclusion, the use of high oxygen tension (20% maintained by exposure to 5% CO2 in air) for IVM of porcine oocytes promoted blastocyst formation in vitro.

Animals↗
PubMed2005

The analysis of telomere length and telomerase activity in cloned pigs and cows.

Inefficiency in the production of cloned animals is most likely due to epigenetic reprogramming errors after somatic cell nuclear transfer (SCNT). In order to investigate whether nuclear reprogramming restores cellular age of donor cells after SCNT, we measured telomere length and telomerase activity in cloned pigs and cattle. In normal pigs and cattle, the mean telomere length was decreased with biological aging. In cloned or transgenic cloned piglets, the mean telomere length was elongated compared to nuclear donor fetal fibroblasts and age-matched normal piglets. In cloned cattle, no increases in mean telomere length were observed compared to nuclear donor adult fibroblasts. In terms of telomerase activity, significant activity was observed in nuclear donor cells and normal tissues from adult or new-born pigs and cattle, with relatively higher activity in the porcine tissues compared to the bovine tissues. Cloned calves and piglets showed the same level of telomerase activity as their respective donor cells. In addition, no difference in telomerase activity was observed between normal and transgenic cloned piglets. However, increased telomerase activity was observed in porcine SCNT blastocysts compared to nuclear donor cells and in vitro fertilization (IVF)-derived blastocysts, suggesting that the elongation of telomere lengths observed in cloned piglets could be due to the presence of higher telomerase activity in SCNT blastocysts. In conclusion, gathering from the comparative studies with cattle, we were able to demonstrate that telomere length in cloned piglets was rebuilt or elongated with the use of cultured donor fetal fibroblasts.

Animals↗
PubMed2005

Cats cloned from fetal and adult somatic cells by nuclear transfer.

This work was undertaken in order to study the developmental competence of nuclear transfer (NT) into cat embryos using fetal fibroblast and adult skin fibroblast cells as donor nuclei. Oocytes were recovered by mincing the ovaries in Hepes-buffered TCM199 and selecting the cumulus oocyte complexes (COCs) with compact cumulus cell mass and dark color. Homogenous ooplasm was cultured for maturation in TCM199+10% fetal bovine serum (FBS) for 12 h and used as a source of recipient cytoplast for exogenous somatic nuclei. In experiment 1, we evaluated the effect of donor cell type on the reconstruction and development of cloned embryos. Fusion, first cleavage and blastocyst developmental rate were not different between fetal fibroblasts and adult skin cells (71.2 vs 66.8; 71.0 vs 57.6; 4.0 vs 6.1% respectively; P < 0.05). In experiment 2, cloned embryos were surgically transferred into the oviducts of recipient queens. One of the seven recipient queens was delivered naturally of 2 healthy cloned cats and 1 stillborn from fetal fibroblast cells of male origin 65 days after embryo transfer. One of three recipient queens was delivered naturally of 1 healthy cloned cat from adult skin cells of female origin 65 days after embryo transfer. The cloned cats showed genotypes identical to the donor cell lines, indicating that adult somatic cells can be used for feline cloning.

Animals↗
PubMed2005

Omental infarction in children: imaging features with pathological correlation.

INTRODUCTION: Omental infarction is a rare occurrence in the paediatric population. It often presents as an acute abdomen that can mimic acute appendicitis and cholecystitis. METHODS: Six cases of omental infarction in children, proven on histopathology, were retrospectively reviewed for their clinical presentation and imaging findings on ultrasonography and computed tomography. RESULTS: These cases revealed clinical and imaging findings on computed tomography that were suggestive and helpful in the pre-operative diagnosis of omental infarction. Findings on ultrasonography were less specific. Histopathological specimens revealed findings of vasculitis in all cases. CONCLUSION: There are clinical and imaging features that will help in the pre-operative diagnosis of this uncommon condition. We also postulate vasculitis as a possible underlying pathology for omental infarction.

Child↗
PubMed2005

Fetus-in-fetu in the pelvis: report of a case and literature review.

INTRODUCTION: Fetus-in-fetu is an extremely rare condition in which a malformed fetus is found in the body of its twin. To our knowledge, fewer than 100 cases have been reported. Wide variations of presentation have been described, although its embryo-pathogenesis and differentiation from a teratoma have not been well established. CLINICAL PICTURE: We describe a male neonate with a fetoid-like mass in his pelvis associated with bilateral undescended testes. The mass was detected on prenatal ultrasound scans. The diagnosis of fetus-in-fetu was considered prenatally and confirmed on a computed tomography scan after birth. OUTCOME: The mass was successfully excised. Histological examination, accompanied by a review of the literature, confirmed that the mass had features consistent with a fetus-in-fetu. CONCLUSIONS: Although an extremely rare clinical entity, fetus-in-fetu can be diagnosed prior to surgery with current imaging modalities. When it arises in the retroperitoneum of a male infant, it can hinder the descent of the testes. Complete excision is curative.

Cryptorchidism↗
PubMed2004

Incidence of apoptosis in clone embryos and improved development by the treatment of donor somatic cells with putative apoptosis inhibitors.

This study was conducted to promote in vitro-development of clone embryos by the treatment of donor somatic cells with hemoglobin (Hb) and/or beta-mercaptoethanol (ME), based on the analysis of apoptosis after somatic cell nuclear transfer (SCNT). Prospective, randomized study was conducted and, in vitro-matured bovine oocytes and fetal fibroblasts were provided for SCNT. In the first series of experiment, embryo apoptosis after SCNT was monitored by a terminal deoxynucleotidyl transferase-mediated d-UTP nick end-labeling assay. As results, apoptosis occurred more (P < 0.05) frequently after SCNT than after in vitro-fertilization (IVF) of control treatment. Subsequently, donor somatic cells treated with Hb (1 microg/ml) and/or ME (10 microM) were provided for SCNT. Either Hb or ME greatly reduced apoptosis (0.083 +/- 0.006 vs. 0.058-0.068 +/- 0.005), while combined treatment did not. ME was more promotive than Hb; significant increases were found in morula compaction (86%), cell numbers of blastocyst (131.3 +/- 1.3 cells/blastocyst), and inner cell mass (31.9 +/- 0.8 cells/blastocyst) cell, and the ratio of inner cell mass to trophectodermal cell numbers (0.24 +/- 0.01). In conclusion, the treatment of donor somatic cells with ME or Hb could reduce apoptosis after SCNT, resulting improved preimplantation development.

Animals↗
PubMed2004

Comparative analysis on the 5'-untranslated region of bovine viral diarrhea virus isolated in Korea.

Samples of 249 bovine abortuses, one intestine, and four diarrheal stools from 254 cows were collected, and bovine viral diarrhea virus (BVDV) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Subsequently, virus isolation was preformed with PCR-positive samples, and then PCR product of 5(')-untranslated region (UTR) of BVDV isolate was sequenced and analyzed. Among the samples collected, 20 (from 17 bovine abortuses, one intestine, and two diarrheal stools) were positive for BVDV RT-PCR; four BVDVs (from two bovine abortuses, one intestine, and one diarrheal stool) were isolated. When the four isolates were biotyped in cell culture, one BVDV isolate from a bovine abortus was cytopathic and the others were non-cytopathic. In addition, three isolates were genotyped as BVDV-1 and one isolate from a diarrheal stool as BVDV-2. In phylogenetic analysis, it suggested that the BVDV-2 isolate in Korea is closer to the North American strains than Asian strains. This is the first report on the identification and isolation of BVDV-2 in Korea.

5' Untranslated Regions↗
PubMed2004

Effects of amino acids on maturation, fertilization and embryo development of pig follicular oocytes in two IVM media.

This study was conducted to develop a serum-free, defined medium for IVM of pig oocytes. Modified North Carolina State University (mNCSU)-23 media with or without supplementation with both epidermal growth factor (EGF) and gonadotrophin were used as base media. In separate experiments, each base medium was supplemented with porcine follicular fluid (pFF), polyvinyl alcohol (PVA), PVA and essential amino acids (EAA), PVA and nonessential amino acids (NEAA) or PVA with both EAA and NEAA. Averaged across these five treatments, the percentage of blastocyst formation was higher (P < 0.05) in the base medium supplemented with EGF and gonadotrophins. In both base media, the addition of NEAA yielded similar percentages of maturation (81-82% versus 75-80%), sperm penetration (89-93% versus 80-86%) and blastocyst formation (4-18% versus 4-13%) as media supplemented with pFF. Although similar benefits were found after the addition of EAA, their addition was associated with lower (P < 0.05) maturation (66%) and sperm penetration (58%) than when pFF was added to the base medium without EGF and gonadotrophins. However, decreased maturation after EAA addition was not detected in the base medium containing EGF and gonadotrophins. Within the same base medium, monospermy, male pronucleus formation, cleavage and blastocyst formation were not affected by the treatments; and combined addition of EAA and NEAA did not further improve oocyte development. In conclusion, a maturation system using a defined mNCSU-23 medium supplemented with EGF, gonadotrophins and EAA or NEAA was developed which yielded a similar number of blastocysts compared with a pFF-containing medium.

Amino Acids↗
PubMed2003

Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos.

In order to develop a culture system and recipient cytoplasm that could improve the developmental competence of somatic cell nuclear transfer (SCNT) embryos for successful cloning of pigs, we evaluated the effect of donor oocytes and in vitro maturation (IVM) media on maturation of oocytes and developmental competence of SCNT embryos. In Experiment 1, oocytes derived from sows or gilts were matured in two IVM media (TCM-199 versus NCSU-23) and maturation of oocytes was evaluated by the status of chromatin configuration, the diameter of matured oocytes, the thickness of the zona pellucida, and the size of the perivitelline space (PVS). Sow oocytes matured in TCM-199 (S-TCM group) and NCSU-23 (S-NCSU group) showed significantly higher (P<0.05) maturation rates (S-TCM and S-NSCU, 86+/-4 and 82+/-4%, respectively) when evaluated by metaphase-II status than the gilt oocytes matured in TCM-199 (G-TCM group, 71+/-3%) and in NCSU-23 (G-NCSU-23 group, 71+/-3%). Oocyte diameter, the thickness of the zona pellucida, and the perivitelline space of sow oocytes (S-TCM and S-NCSU) were larger than those of gilt oocytes (G-TCM and G-NCSU) after IVM (P<0.05). In Experiment 2, SCNT was performed, using in vitro-matured oocytes from each group as recipient cytoplasm and porcine fetal fibroblasts as karyoplasts. The reconstructed embryos were electrically fused and activated, and cleavage and blastocyst formation were monitored under a stereomicroscope. The total cell number of flattened blastocysts stained with 5 microM bisbenzimide on day 7 were counted. In addition, in vitro matured non-enucleated oocytes were also electrically activated (parthenogenetic activation) and pronuclear formation was monitored. No difference in pronuclear formation rate after parthenogenetic activation and fusion rate after SCNT was observed among experimental groups. A significantly higher cleavage rate (P<0.05) was observed in S-TCM (69+/-4%) when compared with only G-NCSU (58+/-4%), but not with G-TCM (60+/-4%) or S-NCSU (68+/-4%). The rate of blastocyst formation was significantly higher (P<0.05) in sow oocytes (24% in S-TCM and S-NCSU), when compared to that observed in G-TCM (15%), and G-NCSU (14%). When the same source of oocytes was used, there was no significant difference in rate of blastocyst formation in the two culture media. Total cell number of blastocysts were not significantly different among experimental groups. In conclusion, the present study clearly demonstrated that sow oocytes have a greater developmental competence than gilt oocytes, regardless of the maturation medium examined.

Animals↗
PubMed2003

Clinics in diagnostic imaging (90). Childhood nasopharyngeal carcinoma.

An 11-year-old boy presented with a nasopharyngeal mass that was thought to represent a juvenile angiofibroma based on the initial clinical and radiological evaluation. Partial tumour resection was performed. Resected specimen revealed histological diagnosis of undifferentiated carcinoma. Further evaluation of the tumour including MR imaging, radioisotope bone scan, CT thorax and abdomen were performed. Differential diagnoses of childhood nasopharyngeal masses were discussed. The differences between childhood NPC and adult NPC, rhabdomyosarcoma, malignant lymphoma and juvenile nasopharyngeal angiofibroma were also discussed.

Carcinoma↗
PubMed2002

Optimization of culture medium for cloned bovine embryos and its influence on pregnancy and delivery outcome.

This study was conducted to establish an effective culture system for supporting in vitro development of cloned bovine embryos and to evaluate whether improved development in the optimal culture system could contribute to enhancing pregnancy and delivery outcomes after transfer. Enucleated oocytes at the metaphase II stage were reconstructed with serum-starved ear fibroblasts and cloned embryos were subsequently cultured for 168 h in vitro. In Experiment 1, cloned embryos were cultured in either modified Charles Rosenkrans 2 amino acid medium (mCR2aa) or modified synthetic oviduct fluid medium (mSOF). More (P < 0.05) 2-cell embryos (78% versus 92%), morulae (51% versus 69%) and blastocysts (2% versus 39%) were obtained after culture in mSOF than after culture in mCR2aa. In Experiment 2, cloned embryos were successively cultured in mSOF supplemented with various macromolecules during different periods of culture. A successive culture of oocytes in BSA-containing medium for 72 h and then in FBS-containing medium for the next 96 h yielded a higher rate of blastocyst formation (49% versus 25-36%) than other combinations (BSA to BSA or PVA to PVA, BSA or FBS). This macromolecule supplementation also significantly increased the number of total blastomeres (117.3 cells/blastocyst) and inner cell mass cells (ICM, 49.7 cells/blastocyst), and the ratio of ICM cells to trophoblast cells (TB, 0.98). In Experiment 3, a total of 85 blastocysts obtained from each 2-step culture were transferred individually to recipient cows at the end of the culture period and 32 pregnancies (38%) were diagnosed on Day 60 after transfer. However, no (P > 0.05) significant differences due to culture were apparent in the pregnancy outcome. Although six calves were produced using the 2-step culture regime of either BSA-BSA or PVA-FBS, no calves were produced using the successive culture of BSA then FBS, which optimized preimplantation development. In conclusion, mSOF has more potential to support the development of clone embryos than mCR2aa, and successive supplementation of BSA and FBS to mSOF further promotes blastocyst formation. However, enhanced development in vitro might not directly contribute to improving pregnancy outcomes.

Animals↗
PubMed2002

Development of bovine oocytes reconstructed with different donor somatic cells with or without serum starvation.

We conducted this study to examine whether serum starvation in culture contributes to better development of bovine reconstructed oocytes and to evaluate which serum-starved somatic cell is the most effective for cloned calf production. In Experiment 1, donor cells of four different types (cumulus cells, ear fibroblasts, oviduct cells and uterine cells) were either serum-starved or not before fusion with enucleated oocytes, and reconstructed oocytes were further cultured for 168 h. Regardless of serum starvation, cumulus cells or ear fibroblasts yielded higher (P < 0.05) rates of fusion than other cells (62.6-69.3 versus 33.3-38.7%). In the serum-starved group, the first cleavage after reconstruction was significantly increased in cumulus cells and ear fibroblasts, compared with oviduct cells (93.4-94.3 versus 78.8-86.0%), and oocytes reconstructed with either of these yielded more blastocysts than oocytes reconstructed with oviduct or uterine cells (40.6-43.8 versus 20.3-19.0%). We observed a similar pattern in the non-starved group, but we found a significant increase in blastocyst formation was found only in cumulus cells compared with other donor cells (42.6 versus 15.4-27.7%). Overall comparison showed that serum starvation increased the rates of cleavage and development to the blastocyst stage, but we found a statistical significance only in the cleavage rate (80.0 versus 89.5%). In Experiment 2, we transferred randomly selected 59 blastocysts that were developed from oocytes reconstructed with serum-starved cells to 44 synchronised recipients. Of those recipients, 23 became pregnant on Day 60 after transfer (52.3%) and 12 (27.3%) delivered cloned calves. The mean gestation length and birth weight was 275 +/- 8 days and 39.6 +/- 15.6 kg, respectively. Although there was no significant difference among donor cells, blastocysts that were derived from oocytes reconstructed with ear fibroblasts yielded the highest rates of pregnancy (50.0%) and delivery (27.3%). In conclusion, serum starvation is effective for improving preimplantation development of oocytes reconstructed with cumulus or ear fibroblast cells and it may positively influence on obtaining better pregnancy outcome.

Animals↗
PubMed2002

Encephalomyelitis associated with akabane virus infection in adult cows.

Between August and September 2000, five 2-7-year-old cows in Korea exhibited neurologic signs and were diagnosed as infected with Akabane virus based on the results of histopathology, immunohistochemistry, serology, and reverse transcription polymerase chain reaction (RT-PCR) analysis. Immunohistochemistry and RT-PCR were equally effective and sensitive for diagnosing Akabane virus infection during the early stage of infection. Typical lymphohistiocytic inflammation characterized by perivascular mononuclear cell infiltration, gliosis, neuronophagia, and neuronal loss was noted in the brain and the ventral horn gray matter of the spinal cord. The lesions in the brain were most prominent in the pons and medulla oblongata. Akabane virus antigen was detected in the brain and spinal cord, mainly in degenerating neurons and glial cells. RT-PCR analysis revealed a target band of expected size in four cows. This is the first report on an outbreak of natural Akabane virus infection in adult cattle.

Animals↗
PubMed2001

A separate procedure of fusion and activation in an ear fibroblast nuclear transfer program improves preimplantation development of bovine reconstituted oocytes.

This study was conducted to examine whether preimplantation development of bovine ("HanWoo," Bos Taurus corcanae) oocytes reconstituted with ear fibroblasts could be improved by a modified procedure of fusion and activation. In Experiment 1, enucleated oocytes were reconstituted with ear fibroblast by a combined procedure of electric fusion and activation at either 24 or 28 hours after IVM. The 28 hours reconstitution yielded more blastocysts (4% vs. 21%, P = 0.0025) and higher ratio of blastocysts per 2-cell embryos (0.05 vs. 0.25, P = 0.003) than the 24 hours. In Experiment 2, enucleated oocytes were reconstituted by one of the three fusion and activation protocols; 1) a combined procedure of electric fusion and activation at 28 hours after IVM, 2) a combined procedure of electric fusion and chemical activation at 28 hours, and 3) a separate procedure of electric fusion at 24 hours and chemical activation at 28 hours. When compared two combined procedures, chemical activation with 5 microM ionomycin for 4 minutes did not promote embryo development and significantly reduced the fusion rate (42% vs. 53%, P = 0.0395). However, significant (P < 0.0113) increases in the development to 2-cell (90% vs. 70 to 74%) and blastocyst (47% vs. 7 to 13%) stages and in the ratio of blastocysts per 2-cell embryos (0.52 vs. 0.11 to 0.18) were obtained by a separate procedure of electric fusion and chemical activation than by the combined procedures. This separate protocol did not reduce the fusion rate compared with the combined procedures (58%). In conclusion, improved development of oocytes reconstituted with ear fibroblasts was achieved by undergoing a separate procedure of electric fusion and chemical activation 4 hours apart.

Animals↗
PubMed2001

Cryopreservation of ICR mouse oocytes: improved post-thawed preimplantation development after vitrification using Taxol, a cytoskeleton stabilizer.

OBJECTIVE: To establish an effective cryopreservation method. DESIGN: In vitro model study. SETTING: Infertility Medical Center, Pochon CHA University. ANIMAL(S): Four-week-old ICR mice superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin. INTERVENTION(S): Vitrified-thawed oocytes were fertilized and subsequently cultured in vitro. MAIN OUTCOME MEASURE(S): Post-thawed development, chromosome/spindle normalities, and blastocyst quality. RESULT(S): More cumulus-enclosed oocytes were fertilized and developed to the 8-cell stage after vitrification and thawing than denuded oocytes. However, cryopreserved oocytes of both types had lower spindle and chromosome normalities than fresh oocytes, which resulted in reduced developmental competence after thawing. The addition of 1 microM of Taxol, a cytoskeleton stabilizer, to vitrification solution greatly promoted the blastocyst formation of vitrified-thawed oocytes, compared with no addition (24.0% vs. 58.6%). No difference in blastocyst quality, which was evaluated by blastomere and inner cell mass cell numbers and inner cell mass cell per trophoblast ratio, was found between fresh oocytes and oocytes vitrified with Taxol. CONCLUSION(S): A vitrification solution consisting of 5.5 M ethylene glycol, 1.0 M sucrose, 10% fetal bovine serum, and 1 microM Taxol greatly improved post-thawed development of vitrified oocytes.

Animals↗