• Development and Reproduction
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• v.12 no.1
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• pp.19-30
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• 2008

Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

• Journal of Animal Science and Technology
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• v.44 no.4
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• pp.391-398
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• 2002

This study was conducted to determine the polymorphism of transferrin exons 13, 15 and 16 by Single-Strand Conformation Polymorphism(SSCP) analysis and to compare their genotypes of Cheju horse Group I (Cheju Institute), Cheju horse Group II (farms), and Thoroughbred (KRA). SSCP of transferrin exon 13, 15, and 16 showed two (A, B), three (A, B, C) and three (A, B, C) codominant alleles, respectively. The Group I and Thoroughbred showed the similar frequencies of allele A and B in transferrin exon 13, but only allele A was observed in Group Ⅱ. In transferrin exons 15 and 16, the frequencies of each allele were different in each Groups. The multiple allele frequencies in exons 15 and 16 suggested that the genotyping of this locus could be used to identify an individual and to test the parentage of offspring. The probability for parentage exclusion were 0.46 and 0.374 for exons 15 and 16 for Cheju horse Group I. Among the 13 combined genotypes of exons 13, 15 and 16, the genotype AA-AB-AB (0.372) is the most common in Cheju horse Group I, but genotype AA-AA-AA is common in the Cheju horse Group II (0.366) and Thoroughbred (0.767). The present study showed two new SNP, which was at the cDNA position 1626 (A/G) in B allele of the exon 13 and 2075 (C/T) in C allele of the exon 16 resulting in amino acid change (Threonine $\longrightarrow$ Methionine). Result showed that polymorphism of exons 13, 15 and 16 in Cheju horses was as high as in Thoroughbred and there was a differences of transferrin allele frequencies in Cheju horses.

• Journal of Korean Academy of Nursing
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• v.5 no.1
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• pp.1-16
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• 1975

The newborn human is the only mammalian whose mother does not have a food supply ready for it's offspring at birth. From two to four days usually elapse before the mother's supply If milk appears, and during this period, some kind of artificial feeding should be supplied to the infants. Because of this factor, there has been continued debate fog the past hundreds of years as to when the first feeding should be started Accordingly, many experiments were carried out by scholars and because of these, Pre-lateral feedings were believed to be necessary. Many types of pre-lateral feedings were tried and the conclusion was reached that glucose water was the best food for the first infants'feedings. Traditionally, This has been started 12 hours after birth. The causes for the 12 hours delay were thought to (1) provide rest for the infants: (2) prevent regurgitation ana vomiting which tended to be prevalent during this tine: (3) in cases of low weight infants, prevention of aspiration pneumonia. From recent studies of newborn physiology and as pediatric medicine has been rapidly advancing, many studies hare been carried out concerning the improvement of infant nutrition and the early feeding of infants has been emphasized. This author believes it would be very beneficial to try two different kinds of feedings for the infant. (1) experimental feedings ana (2) comparative feeding, and during this period to investigate and compare the infants blood sugar level, hematocrit, gamma globulin level weight changes and to observe the infant reaction ill order to search for a more desirable feeding program. This study was conducted from January to March 1974 with data related to 40 healthy newborn infants (male 21, female 19: weight, 2.79∼4.20㎏ : gestation, 39∼40 weeks) born at Ewha Womens University Hospital and the results obtained were as follows : 1. At time of birth the blood sugar level from the cord sample averaged 88.99㎎/100㎖, but the blood sugar level rapidly dropped after 2 to 3 hours and reached the lowest point after 10 to 11 hours (54.48㎎/100㎖) and rose again by the 24 hour time period (76.80㎎/100㎖). Changes in the blood sugar level of the experiments: groups and the compare-five group was not significantly different until the 6 to 7 hour period, but by the 10 to 11 hour period the blood sugar levels of the experimental group (49,10㎎/100㎖) and the comparative group (49.70㎎/100㎖) were lower than the remainder of the experimental groups. 9. There ware no significant weight changes between the two groups. Average weight at birth was 3.35㎏, but at the 24 hours period birth weight averaged 3.29㎏. (1.8% reduction of birth weight). It continually lowered until at 48 hours, average weight was 3.26㎏ (2.7% reduction from birth weight.) 3. Hematocrit readings showed no significant difference between the groups. Hematocrit, the average value at birth, was 28.07% and abruptly elevated to average 64.35% at the 2 to 3 hour period, then slowly lowered to an average of 59.67% at the 6 to 7 hour period, 55.10% at the 10 to 11 hour period, ana 53.70% at the 24 hour period. 4. At birth, average gamma globulin value averaged 1,39㎎/100㎖. and at the 24 hour period averaged 1,52㎎/100㎖ revealing no significant difference between the two feeding groups. 5. Such factors as voiding, passing of meconium, regurgitation and vomiting showed no significance between the two feeding groups. However, the number of infants voiding and passing meconium in the experimental groups during the first 12 hours was slightly greater. In general there was an increased tendency for regurgitation and. vomiting among a small group of the infants during the first 24 hours which thereafter decreased. 6. Fluid intake averaged 24.38cc at the first feeding and increased to average 30.48cc at the third feeding and further increased to 73. 00cc at the fifteenth feeding. Finally it was suggested that the most reasonable method of early feeding is to give less than 25cc of 5% glucose water and/or 8% powdered milk at 8 to 9 flours after birth in order to prevent hypoglycemia and dehydration.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.135-142
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• 2001

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.105-110
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• 2004

Objectives: Despite severe oligospermia, males with Y chromosome microdeletion can achieve conception through ICSI (Intracytoplasmic Sperm Injection). However, ICSI may not only result in the transmission of microdeletions but also the expansion of deletion to the offspring. The purpose of this study was to screen vertical transmission, expansion of microdeletions and de novo deletion in male fetuses conceived by ICSI. Materials and Methods: A total of 32 ICSI treated patients with their 33 (a case of twin) male fetuses conceived by ICSI were used to make this study group. Sequence-tagged sites (STSs)-based PCR analyses were performed on genomic DNA isolated from peripheral blood of fathers and from the amniocytes of male fetuses. Ten primer pairs namely, sY134, sY138, MK5, sY152, sY147, sY254, sY255, SPGY1, sY269 and sY158 were used. The samples with deletions were verified at least three times. Results: We detected a frequency of 12.5% (4 of the 32 patients) of microdeletions in ICSI patients. In 4 patients with detected deletions, two patients have proven deletions on single STS marker and their male fetuses have the identical deletion in this region. Another two patients have two and three deletions, but their male fetuses have more than 3 deletions which include deletions to their father's. Meanwhile, seven male fetuses, whose fathers were analyzed to have all 10 STS markers present, have deletions present in at least one or more of the markers. Conclusions: Although the majority of deletions on the Y chromosome are believed to arise de novo, in some cases a deletion has been transmitted from the fertile father to the infertile patient. In other cases the deletion was transmitted through ICSI treatment, it is likely that one sperm cell is injected through the oocyte's cytoplasm and fertilization can be obtained from spermatozoa. Our tests for deletion were determined by PCR and our results show that the ICSI treatment may lead to vertical transmission, expansion and de novo Y chromosome microdeletions in male fetuses. Because the sample group was relatively small, one should be cautious in analyzing these data. However, it is important to counsel infertile couples contemplating ICSI if the male carries Y chromosomal microdeletions.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.9
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• pp.4517-4525
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• 2016

Background: Sperm DNA damage is underlying aetiology of poor implantation and pregnancy rates but also affects health of offspring and may also result in denovo mutations in germ line and post fertilization. This may result in complex diseases, polygenic disorders and childhood cancers. Childhood cancer like retinoblastoma (RB) is more prevalent in developing countries and the incidence of RB has increased more than three fold in India in the last decade. Recent studies have documented increased incidence of cancers in children born to fathers who consume alcohol in excess and tobacco or who were conceived by assisted conception. The aetiology of childhood cancer and increased disease burden in these children is lin ked to oxidative stress (OS) and oxidative DNA damage( ODD) in sperm of their fathers. Though several antioxidants are in use to combat oxidative stress, the effect of majority of these formulations on DNA is not known. Yoga and meditation cause significant decline in OS and ODD and aid in regulating OS levels such that reactive oxygen speues meditated signal transduction, gene expression and several other physiological functions are not disrupted. Thus, this study aimed to analyze sperm ODD as a possible etiological factor in childhood cancer and role of simple life style interventions like yoga and meditation in significantly decreasing seminal oxidative stress and oxidative DNA damage and thereby decreasing incidence of childhood cancers. Materials and Methods: A total of 131 fathers of children with RB (non-familial sporadic heritable) and 50 controls (fathers of healthy children) were recruited at a tertiary center in India. Sperm parameters as per WHO 2010 guidelines and reactive oxygen species (ROS), DNA fragmentation index (DFI), 8-hydroxy-2'-deoxy guanosine (8-OHdG) and telomere length were estimated at day 0, and after 3 and 6 months of intervention. We also examined the compliance with yoga and meditation practice and smoking status at each follow-up. Results: The seminal mean ROS levels (p<0.05), sperm DFI (p<0.001), 8-OHdG (p<0.01) levels were significantly higher in fathers of children with RB, as compared to controls and the relative mean telomere length in the sperm was shorter. Levels of ROS were significantly reduced in tobacco users (p<0.05) as well as in alcoholics (p<0.05) after intervention. DFI reduced significantly (p<0.05) after 6 months of yoga and meditation practice in all groups. The levels of oxidative DNA damage marker 8-OHdG were reduced significantly after 3 months (p<0.05) and 6 months (p<0.05) of practice. Conclusions: Our results suggest that OS and ODD DNA may contribute to the development of childhood cancer. This may be due to accumulation of oxidized mutagenic base 8OHdG, and elevated MDA levels which results in MDA dimers which are also mutagenic, aberrant methylation pattern, altered gene expression which affect cell proliferation and survival through activation of transcription factors. Increased mt DNA mutations and aberrant repair of mt and nuclear DNA due to highly truncatred DNA repair mechanisms all contribute to sperm genome hypermutability and persistant oxidative DNA damage. Oxidative stress is also associated with genome wide hypomethylation, telomere shortening and mitochondrial dysfunction leading to genome hypermutability and instability. To the best of our knowledge, this is the first study to report decline in OS and ODD and improvement in sperm DNA integrity following adoption of meditation and yoga based life style modification.This may reduce disease burden in next generation and reduce incidence of childhood cancers.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.7
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• pp.925-937
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• 2016

In the last few decades, transgenic animal technology has witnessed an increasingly wide application in animal breeding. Reproductive traits are economically important to the pig industry. It has been shown that the bone morphogenetic protein receptor type IB (BMPR1B) A746G polymorphism is responsible for the fertility in sheep. However, this causal mutation exits exclusively in sheep and goat. In this study, we attempted to create transgenic pigs by introducing this mutation with the aim to improve reproductive traits in pigs. We successfully constructed a vector containing porcine BMPR1B coding sequence (CDS) with the mutant G allele of A746G mutation. In total, we obtained 24 cloned male piglets using handmade cloning (HMC) technique, and 12 individuals survived till maturation. A set of polymerase chain reactions indicated that 11 of 12 matured boars were transgene-positive individuals, and that the transgenic vector was most likely disrupted during cloning. Of 11 positive pigs, one (No. 11) lost a part of the terminator region but had the intact promoter and the CDS regions. cDNA sequencing showed that the introduced allele (746G) was expressed in multiple tissues of transgene-positive offspring of No.11. Western blot analysis revealed that BMPR1B protein expression in multiple tissues of transgene-positive $F_1$ piglets was 0.5 to 2-fold higher than that in the transgene-negative siblings. The No. 11 boar showed normal litter size performance as normal pigs from the same breed. Transgene-positive $F_1$ boars produced by No. 11 had higher semen volume, sperm concentration and total sperm per ejaculate than the negative siblings, although the differences did not reached statistical significance. Transgene-positive $F_1$ sows had similar litter size performance to the negative siblings, and more data are needed to adequately assess the litter size performance. In conclusion, we obtained 24 cloned transgenic pigs with the modified porcine BMPR1B CDS using HMC. cDNA sequencing and western blot indicated that the exogenous BMPR1B CDS was successfully expressed in host pigs. The transgenic pigs showed normal litter size performance. However, no significant differences in litter size were found between transgene-positive and negative sows. Our study provides new insight into producing cloned transgenic livestock related to reproductive traits.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.9
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• pp.432-439
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• 2020

To improve the relatively low economic efficiency of the Korean native pig, the Korean National Institute of Animal Science developed a novel pig breed, the Woori black pig (W), by crossing Korean native and Duroc (D) pigs. This study was conducted to evaluate the effects of W as a terminal sire on growth performance, body shape, and retail cut yield of crossbred pigs. By using a completely randomized design, 32 crossbred pigs were allotted to one of two treatment groups based on terminal sire. The two groups were LYD [(Landrace × Yorkshire) × D sire] and LYW [(Landrace × Yorkshire) × W sire]. The experimental assessments were conducted over 53 days. The terminal sire breed had no significant effect on body weight (BW) at d 53, or on BW gain, average daily gain, or days to reach a 90 kg body weight. Moreover, there were no significant differences in body length, body height, or chest depth between the two groups. However, there was a significant difference (p < 0.05) in backfat thickness between the LYD (17.29 mm) and LYW (18.96 mm) groups. Loin yield of crossbred pigs in the LYW group (13.11%) was significantly lower (p < 0.05) than that in the LYD group (13.85%). By contrast, the Boston butt yield was significantly higher (p < 0.05) in the LYW group (8.99%) than in the LYD group (8.21%). In conclusion, these results suggest crossbred pigs sired by a Woori black pig had growth performance, shape, and retail cut yield (except loin yield) Ed. Note: I assume the lower loin yield is a negative factor so I included this wording. similar to those sired by a Duroc pig. The results showed no overall negative effect Ed. Note: I assume the lower loin yield is a negative factor so I used this wording. on crossbred offspring, indicating the suitability of the Woori black pig as a terminal sire.

• Journal of Animal Science and Technology
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• v.49 no.6
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• pp.711-718
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• 2007

Most cattle breeds have a coat color pattern that is characteristic for the breed. Korean cattle(Hanwoo) has a coat color ranging from yellowish brown to dark brown including a red coat color. Variation in the Hanwoo coat color is likely to be the effects of modified genes segregating within the Hanwoo breed. MC1R encoded by the Extension(E) locus was almost fixed with recessive red e allele in the Hanwoo, but other gene(s) might be affecting the variation of the Hanwoo coat color into yellowish to red brown. We have analyzed a segregation of coat color in the F2 families generated from two Hanwoo bulls(yellowish brown) mated to six F1 dams(black) derived from Hanwoo and Holstein crosses. Segregation of coat color in the offspring found a ratio of 1(yellowish brown) : 1(black) and this ratio indicates that a single gene may play a major role for the Hanwoo coat color. We further investigated SNPs in MC1R, ASIP and TYRP1 loci to determine genetic cause of the Hanwoo coat color. Several polymorphisms within ASIP intron 2 and TYRP1 exons were found but not conserved within the Hanwoo population. However, the segregation of the MC1R e allele was completely associated with the Hanwoo coat color. Based on this information, it is clear that the MC1R e allele is mainly responsible for the yellowish red Hanwoo coat color. Further study is warrant to identify possible genetic interaction between MC1R e allele and other coat color related gene(s) for the variation of Hanwoo coat color from yellowish brown to dark brown. (Key words : Hanwoo, Coat color, SNP, MC1R, ASIP, TYRP1)

• Journal of Embryo Transfer
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• v.18 no.2
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• pp.97-108
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• 2003

In-vitro fertilized Hanwoo embryos were biopsied for sex determination by PCR. Biopsied embryos were incubated for 1∼2 hours for the recovery. Those sexed Hanwoo embryos were transferred to 49 Hanwoo and 16 Holstein recipients from February 2000 to February 2001. Of 65 recipients, 14 cows(12 Hanwoo and 2 Holstein) delivered the same offspring as sex-determined by PCR, therefore the conception rate was 21.5%. 1. Total 65 embryos(male 35, female 30) were transferred to recipients, and 14 calves (male 6, female 8) were delivered. In comparison between sex by PCR method and sex of calves born after embryo transfer, the accuracy of sex determination was 100.0%. 2. The conception rate after transfer with biopsied embryo between Hanwoo and Holstein was 24.5% and 12.5% 3. The conception rate after transfer with biopsied embryo between fresh and frozen-thawed embryos was 23.5% and 14.3%. 4. The conception rate according to the season when embryo was transferred was 11.8, 29.4, 23.5 and 20.0% for spring, summer, autumn and winter, respectively. 5. The conception rate according to embryo quality after biopsy was 41.7, 30.0 and 0.0% for excellent, good and fair quality. 6. The conception rate according to thickness of uterine horn was 71.4, 18.9, 11.8 and 0.0% for 0, +, ++ and +++ thickness. 7. The conception rate according to the site in the uterine hem where embryo was put was 30.0, 20.0 and 10.0% for cranial, mid, and caudal part of uterine horn. 8. The conception rate according to the quality of corpus luteum ipsilateral to the uterine horn where embryos was transferred was 41.2, 14.3 and 15.4% for excellent, good and fair quality. 9. The conception rate according to the time required for embryo transfer was 18.2, 30.0, 30.0, 0.0 and 25.0% for 10, 15, 20, 25 and 30 minutes. 10. The conception rate according to parity of recipients was 26.5, 19.1, 14.3 and 0.0% for the primiparous, the 2nd parous, the 3rd parous and the 4th parous recipients. These results indicated that fresh embryos are more demanded than frozen-thawed embryos for good conception rate in embryo transfer with biopsied-sexed embryo. Also, it was indicated that we should consider embryo-recovering condition, recipient's uterine thickness, transfer site in uterine horn, quality of corpus luteum, time required for transfer and parity of recipient to achieve good conception rate in ET with biopsied-sexed embryos.