• Journal of Embryo Transfer
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• v.25 no.3
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• pp.171-177
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• 2010

Plasminogen activators (PAs) are serine proteases that convert plasminogen to plasmin. The PA/plasmin system has been associated with a number of physiological processes such as fibrinolysis, ovulation and fertilization. Although correlations have been reported between reactive oxygen species (ROS) and oocyte maturation, the relationship between PA activity and ROS is unknown. The present study was undertaken to determine the effects of cumulus cells on PA activity in matured porcine oocytes under xanthine (X)-xanthine oxidase (XO) system. When oocytes were matured under the X-XO system, the proportion of oocytes remaining GV stage was higher (p<0.05) in oocytes without cumulus cells. The incidence of degenerated oocytes was higher (p<0.05) in the X+XO ($11.1{\pm}6.1$ and $21.6{\pm}3.4%$) than in the control group ($2.9{\pm}1.8$ and $4.0{\pm}1.6%$). The proportion of TUNEL-positive oocytes and activity of caspase-3 were higher (p<0.05) in cumulus-free oocytes and oocytes exposed to ROS. Tissue-type plasminogen activator-plasminogen activator inhibitor (tPA-PAI) and tissue-type plasminogen activator (tPA) activity were detected in oocytes that were separated from cumulus-oocytes complexs (COCs) at 44 h of maturation culture, and only tPA was produced in oocytes that were denuded before the onset of maturation culture. On the other hand, the activities of PA were increased (p<0.05) when oocytes were cultured under the X-XO system. The higher activity of tPA was observed in denuded oocytes (DOs) underwent apoptotic changes by oxidative stress. In COCs, however, tPA-PAI as well as tPA activity was detected and apoptotic changes such as DNA cleavage or caspase-3 activation were not observed. These results suggest that tP A may be relevant to apoptotic cell death in porcine oocytes by oxidative stress.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.4
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• pp.331-336
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• 2005

Objective: To investigate the effects of female age on in vitro maturation and fertilization of immature oocytes from controlled ovarian hyperstimulation (COH) in human IVF-ET program. Method: A total of 96 immature oocytes (GV & metaphase I) obtained from 40 cycles of IVF-ET (29 patients). The mean age of female patients was $31.8{\pm}3.1years$. Ovulation was triggered by urinary or recombinant hCG. Immature oocytes were cultured with YS medium containing 30% of patients' human follicular fluids, LH (1 IU/mL), FSH (1 IU/mL) and EGF (10 ng/mL), and then matured oocytes were fertilized by ICSI. In vitro maturation and fertilization of immature oocytes were analyzed according to age of female (< 34 or ${\geq}34years$). Results: The maturation rate was similar between two groups (68% vs 64%). The fertilization rate of in?vitro-matured oocytes was higher in patients < 34 years old, but there was no statistical significance (64% vs 50%, p=0.347). The fertilization rate of in-vitro-matured oocytes was significantly lower compared with those of in-vivo-matured oocytes in both age groups (64% vs 79%, p=0.035, 50% vs 86%, p=0.007). Conclusion: In older female group, fertilization rate of in-vitro-matured oocytes seems to be decreased. Further investigations should be warranted to increase fertilization potential of in-vitro-matured oocytes.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.122-122
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• 2003

This study examined the viability of canine oocytes following storage at 4 or $38{\circ}C$ for 5 hrs. The ovaries were collected from domestic dog following ovariohysterectomy at a local veterinary clinics and transported to laboratory In two different transport temperature at 4 or $38{\circ}C$ within 5 hrs. The cumulus oocyte complexes (COCs) were recovered after slicing with blade. In Exp. 1, the oocytes collected were matured in DMEM supplemented with l0% FBS, 0.6 mM/mlcysteine, 0.2 mM Pyruvic acid, 20 ng/ml $E_2$ and 1 $\mu g/ml$ rbST at humidified atmosphere containing 5% $CO_2 38{\circ}C$ for 24 or 48 hrs to analysis of survivability. In Exp 2, to assess nuclear development at 38{\circ}C$ group, the oocytes were matured in maturation medium for 24, 48 or 96 hrs. Survivability was judged by a morphological appearance and PI staining. Survivability rates were analyzed by General Linear Models procedure in SAS. The survival rates at 4{\circ}C$ ovary transport group showed significantly lower than at 38{\circ}C$ group (0 vs 72.9% in 48 hrs and 13.2 vs 77 8% in 24 hrs; P<0.05). The nuclear development of oocytes to MI to MII stages at 24, 48 and 96 hrs was 8.3% (6/72), 8.9% (9/101), and 9.5% (8/84). These results showed that the canine oocytes were remarkably sensitive to a low temperature and did not increase nuclear development rate depend on maturation time to 96 hrs.

• Journal of the korean academy of Pediatric Dentistry
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• v.37 no.3
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• pp.359-366
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• 2010

It was easy to find that children of a skeletal anterior crossbite in the early mixed dentition period showed a stark difference in the dental maturity between their maxillary and mandibular teeth, if they have stronger physical characteristics. If the difference of dental age between maxillary and mandibular teeth which can be identified via panoramic radiographs may serve as an early sign of class III malocclusion, this is considered valuable as a tool of early detection diagnosis. We obtained lateral cephalometric radiographs, panoramic radiographs, working model and clinical images of patients of Hellman dental age IIA and IIC who visited the department of pediatric dentistry, Pusan National University Dental Hospital and examined them to select 50 patents for normal occlusion group and skeletal anterior crossbite group, respectively. Their panoramic radiographs were used for the Demirjian's method to figure out dental ages of maxillary and mandibular teeth of each group and the eruption rate of the first molars. Their differences are as follows: 1. In both groups, the dental ages from Demirjian's method were advanced than the chronological ages. No sexual dimorphism was detected for the chronological or dental age in either group (p>0.05). 2. The difference of dental age of maxillary and mandibular teeth between the normal occlusion group and crossbite group was 0.22 and 0.69 years, respectively, with a higher difference in crossbite group(p<0.05). 3. Compared to the normal occlusion group, the crossbite group showed a higher difference in the eruption rate between maxillary and mandibular first molar(p<0.05).

• Journal of dental hygiene science
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• v.8 no.2
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• pp.89-96
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• 2008

Effects of sodium fluoride exposure on the amelogenesis during fetal formation were investigated using 11 days rat incisor of control group and two experimental groups. According to results of morphological analysis using an electron microscope, enamel organ in the rat incisor consisted of presecretory, secretory, and maturation zone, especially maturation zone had ruffle-ended ameloblasts (rAB) that additionally supply inorganic ions and smooth-ended ameloblasts (sAB) that remove water and organic compounds. Such a histological composition was same in fetal and adult rats. According to experimental results using calcein (green fluorescence) in order to reveal the modulation cycle of ameloblast, modulation cycle of experimental group decreased on an average one time than control group, as increase of density of sodium fluoride indicated that thickness of smooth-ended ameloblast decreased. Also ratio of thickness on sagittal total length of sAB increased than rAB in experimental groups than control group. In total length of teeth, an injected 100 ppm sodium fluoride group was similar control group but as injected 200 ppm group became short. In experimental group, thickness of sAB and rAB became narrow to the tip of cutting edge. According to concentration of sodium fluoride grows, the modulation cycle and total length of teeth were decreased, finally it prevented teeth growth.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.4
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• pp.317-326
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• 2006

Purpose : The aim of this study was to evaluate the effect of recombinant human bone morphogenetic protein-7 (rhBMP-7) injected on the rate of new-bone formation for distraction osteogenesis on dogs. Materials & Methods : Twelve adult dogs were randomly selected into two groups of six dogs on each. Unilateral osteotomies were performed on the body of the mandible and an intraoral distractor was mounted to the mandible on dogs. One group was treated with injection of rhBMP-7 and the other group served as the control. RhBMP-7 was administered on the day of surgery by single injection into the medullary bone at the osteotomy gap. Distraction was performed five days after osteotomy as a rate of 0.5 mm twice per day for 10 days. The animals were then sacrificed at 2, 4, and 8 weeks after completion of the distraction. Two dogs in each group, totaling four dogs, were killed at 2 weeks, 4 weeks, and 8 weeks after completion of distraction, respectively. The lengthened mandibles were harvested and processed for radiographic and histological examinations. In addition, immunohistochemical examination using osteocalcin expression was studied. Results : Radiographs showed accelerated regenerate ossification with maturation of new bone in the rhBMP-7 group comparing with the control group at the 4 weeks of the consolidation. There was no significant difference in the radiographic findings at the 2 weeks and 8 weeks of the consolidation period. Histological findings demonstrated increased bone healing pattern in the rhBMP-7-treated group during all observation period. The expression of osteocalcin immunoreactivity was hardly detected in the normal mandible of dog, but the expression was detected in all experimental rhBMP-7 treated specimens. There were also significant increasing in number of positive immunostaining cells and staining intensity of osteocalcin expression in the rhBMP-7 treated group compared with those of the control group on 2-weeks and 4-weeks. There was a significant decreasing in staining intenstiy of all both two groups on 8 weeks of consolidation period, but significant differences of immunostaining was not seen in two groups. Conclusions : A single injection of rhBMP-7 at the time of osteotomy may stimulate the rate of regenerate ossification and increase callus maturation during distraction osteogenesis. In addition, it may shorten the distraction osteogenesis procedure and decrease the prevalence of complications associated with mandibular distraction osteogenesis.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.9
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• pp.1227-1231
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• 2004

Pig oocytes contain high levels of lipids in the ooplasm, which reduces the visibility of the germinal vesicle (GV) under microscopic examination. Therefore, the purposes of this study were to investigate the effects of relative centrifugation force (RCF) on the visibility and maturation rates of the GV stage oocytes after centrifugation. In Experiment 1, cumulus-oocyte-complexes (COCs) were collected from slaughterhouse ovaries and randomly allocated to different RCFs (3,000 rpm: 970 g; 6,000 rpm: 3,900 g; or 10,000 rpm: 10,840 g) for 10 or 20 min. Percentages of visible GV were 76-79% in the oocytes centrifuged with 10,000 rpm, which were significantly higher (p<0.01) than those with 3,000 and 6,000 rpm. No significant differences in GV visibility were observed among oocytes with different lengths of centrifugation (p<0.05) regardless of the RCFs. In esperiment 2, the maturation rate of the oocyte was found significantly lower in the 20 min than in the 10 min group received 10,840 g of RCF (30 vs. 75%, p<0.05). In conclusion, the GV of porcine oocytes can be clearly visible by centrifugation at 10,840 g for 10 min without compromising their subsequent maturation rates and a longer centrifugation time (20 min) had no beneficial influence on the visibility of GV stage pig oocytes.

• Journal of Animal Science and Technology
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• v.47 no.3
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• pp.363-370
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• 2005

Experiments were conducted to determine the effects of beta-mercaptoethanol ($\beta$-ME) supplements to the in vitro maturation (IVM) medium on in vitro fertilization (IVF) and intracellular glutathione (GSH) concentration. Porcine cumulus-intact oocytes were matured in TCM-I99 medium containing porcine follicular fluid, sodium pyruvate, D-glucose, FBS, hormonal supplements, and $\beta$-ME (0, 25, 50 and 100 ${\mu}$M) for 36 to 46h. After culture, cumulus-free matured oocytes were co-incubated with epididymal spermatozoa for 18h. There were no significant differences in the maturation rate among treatment groups. However, increases (P < 0.05) in intracellular GSH concentration before and after. fertilization were observed in 50 ${\mu}$M $\beta$-ME supplements to the IVM medium. Also, increases (P < 0.05) in male pronuclear formation after IVF were observed in same treatment group. In conclusion, supplementing $\beta$-ME into the IVM medium increased intracellular GSH concentrations and increased fertilization in vitro.

• The Journal of Korean Obstetrics and Gynecology
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• v.23 no.2
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• pp.57-70
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• 2010

Purpose: In this study, we investigated the effects of GDB(Gidaebang) on the immune response to establish the treatment mechanism of vaginitis. Methods: We examined the effects of GDB on the DCs(Dendritic cells) phenotypic and functional maturation. iDCs were cultured in the presence of GM-CSF and the generated iDCs were respectively stimulated by GDB or LPS as the control group for 24 hours. To evaluate the DCs phenotypic and functional maturation, we used flow cytometric analysis, RT-PCR and ELISA. Results: 1. GDB upregulated the expression of class II MHC and CD40 on DCs. 2. GDB upregulated the expression of CD80 and CD86 on DCs. 3. GDB induced cytokine IL-12 production and mRNA expression in DCs. Conclusion: These results suggest that GDB is able to improve the antigen-presenting capacity of DCs through the upregulation of their maturation, and might induce proliferation of T cells. In conclusion, this immunomodulatory properties of GDB may be useful in the treatment of vaginitis.

• Journal of Ginseng Research
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• v.29 no.4
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• pp.159-166
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• 2005

Ginsenosides, a group of Panax ginseng saponins, exert the lowering effects of plasma cholesterol levels in animals. We had reported earlier that ginsenoside Rb2 upregulate low-density lipoprotein receptor (LDLR) expression via a mechanism that is dependent of the activation of sterol response element binding protein 2 (SREBP-2) expression. This study was conducted to determine the effects of ginsenoside Rb2 on the expression of the hepatic LDLR expression at cellular levels using HepG2 cells, and to evaluate whether the sterol response element binding protein 1 (SREBP-l) was involved in the regulation of LDLR expression. Incubation of HepG2 cells in serum-free medium supplemented with cholesterol $(10{\mu}g/ml)$ for 8 hours decreased the mRNAs of LDLR mRNA by $12\%$ and SREBP-l mRNA by $35\%$. Ginsenoside Rb2 antagonized the repressive effects of cholesterol and increased both LDLR and SREBP-l mRNA expression to 1.5- and 2-fold, respectively. Furthermore, Western blot and confocal microscopic analyses with SREBP-l polyclonal antibody revealed that ginsenoside Rb2 enhanced the maturation of the SREBP-1 from the inactive precursor form in ER membrane to the active transcription factor form in nucleus. These results suggest that ginsenoside Rb2 upregulates LDLR expression via a mechanism that is dependent of the activation of not only SREBP-2 expression, but also SREBP-1 expression and maturation, and also indicate that the pharmacological value of ginsenoside Rb2 may be distinguished from that of lovastatin which is reported that it upregulate LDLR through SREBP-2 only, not through SREBP-1.