• Development and Reproduction
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• v.13 no.1
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• pp.25-33
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• 2009

The cycle of the seminiferous epithelium and development of spermatids of Apodemus speciosus peninsulae were observed using a light microscope. On the basis of developing spermatocyte and spermatid, the cycle of the seminiferous epithelium was divided into 9 stages. Type Ad spermatogonia were appeared in all stages ($I{\sim}IX$). The Ap, In, and B types of spermatogonia were appeared from stage I, II and III, and IV, respectively. In prophase of first meiosis, the leptotene spermatocytes appeared from stage V and VI, zygotene spermatocytes from stages I, II, VII, VIII, and IX, pachytene spermatocytes from stage III to VII, diplotene spermatocytes in the stage VIII, and secondary spermatocytes in stage IX. On the basis of morphology of spermatid head, developing of nuclear and acrosome and the morphological change of cytoplasm, the developing of spermatids was divided into 12 steps. Considering all the results, A. s. peninsulae displayed very similar result with A. agrarius coreae that is allied species when compare correct characters developing of spermatids with spermatogonia and appearance time of the spermatocyte. Appearance time of the same cell and number of spermatogonial generation was thought that characters of the species, and information may be useful in identifying the species.

• Journal of the korean academy of Pediatric Dentistry
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• v.46 no.3
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• pp.239-246
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• 2019

The purpose of this study was to examine dental complications and to evaluate the effects of initial treatment age, treatment modalities, and treatment duration on the disorder after radiation and chemotherapy in pediatric cancer patients. For 93 children with clinical and radiographic data, the number of teeth, the morphology of teeth, the shape of the roots, and development status of the dentition were evaluated. Dental development disorders were found in 61.3% of the children. The mostly found abnormality was root deformity with the prevalence of 31.2%. In children submitted to the therapy before the age of 6, the number of missing teeth (p = 0.029) and microdontia (p = 0.003) were greater compared to the children who started to receive the treatment after the age of 6. The combination of radiation and chemotherapy showed significantly greater incidences of missing teeth (p = 0.030), microdontia (p = 0.046), and root deformity (p = 0.009) when compared with the sole application of chemotherapy. When the children were submitted to anticancer therapy for 18 months or longer duration, greater number of missing teeth (p = 0.032), microdontia (p = 0.011), root deformity (p = 0.025), and total number of teeth affected (p = 0.036) were observed compared with duration less than 18 months. The number of dental abnormalities increased when the children were treated at earlier ages, with combination of radiation and chemotherapy, and for longer period of time.

• Journal of Animal Reproduction and Biotechnology
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• v.34 no.1
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• pp.10-19
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• 2019

Morphology of cumulus-oocyte-complexes (COCs) at germinal vesicle (GV) stage as one of the evaluation criteria for oocyte maturation quality after in vitro maturation (IVM) plays important roles on the meiotic maturation, fertilization and early embryonic development in pigs. When cumulus cells of COCs are insufficient, which is induced the low oocyte maturation rate by the increasing of reactive oxygen species (ROS) in porcine oocyte during IVM. The ROS are known to generate including superoxide and hydrogen peroxide from electron transport system of mitochondria during oocyte maturation in pigs. To regulate the ROS production, the cumulus cells is secreted the various antioxidant enzymes during IVM of porcine oocyte. Our previous study showed that Mito-TEMPO, superoxide specific scavenger, improves the embryonic developmental competence and blastocyst formation rate by regulating of mitochondria functions in pigs. However, the effects of Mito-TEMPO as a superoxide scavenger to help the anti-oxidant functions from cumulus cells of COCs on meiotic maturation during porcine oocyte IVM has not been reported. Here, we categorized experimental groups into two groups (Grade 1: G1; high cumulus cells and Grade 2: G2; low cumulus cells) by using hemocytometer. The meiotic maturation rate from G2 was significantly (p < 0.05) decreased (G1: $79.9{\pm}3.8%$ vs G2: $57.5{\pm}4.6%$) compared to G1. To investigate the production of mitochondria derived superoxide, we used the mitochondrial superoxide dye, Mito-SOX. Red fluorescence of Mito-SOX detected superoxide was significantly (p < 0.05) increased in COCs of G2 compared with G1. And, we examined expression levels of genes associated with mitochondrial antioxidant such as SOD1, SOD2 and PRDX3 using a RT-PCR in porcine COCs at 44 h of IVM. The mRNA levels of three antioxidant enzymes expression in COCs from G2 were significantly (p < 0.05) lower than COCs of G1. In addition, we investigated the anti-oxidative effects of Mito-TEMPO on meiotic maturation of porcine oocyte from G1 and G2. Meiotic maturation and mRNA levels of antioxidant enzymes were significantly (p < 0.05) recovered in G2 by Mito-TEMPO ($0.1{\mu}M$, MT) treatment (G2: $68.4{\pm}3.2%$ vs G2 + MT: $73.9{\pm}1.4%$). Therefore, our results suggest that reduction of mitochondria derived superoxide by Mito-TEMPO may improves the meiotic maturation in IVM of porcine oocyte.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.281-286
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• 2018

Zona pellucida (ZP), a primarily representative coat of mammalian egg and embryo, has an extremely heterogeneous morphology during different developmental stages. The objective of the present study was to compare the morphological changes of the ZP surface of immature, in vitro and in vivo matured canine oocytes by using scanning electron microscopy (SEM). Canine ovaries were collected from local veterinary hospitals to recover immature oocytes. The ovaries were sliced and the released cumulus oocyte complexes (COCs) were washed with TL-HEPES. The selected COCs were randomly divided into two groups, first group was processed immediately at immature state and the second group was processed 72 h after in vitro maturation, and compared with in vivo derived oocytes. Oocytes were fixed, critical point dried and examined under SEM. The diameters of oocyte and outer holes of the ZP were measured on a total of 249 oocytes; the results were analyzed using One-way ANOVA. Our results showed that, the diameter of immature oocytes significantly differed (p < 0.05) from that of in vivo matured oocytes ($79.60{\pm}0.77{\mu}m$ vs. $101.46{\pm}1.07{\mu}m$, respectively). Similarly, a significant difference (p < 0.05) in the diameters between those of in vitro and in vivo matured oocytes were found ($79.51{\pm}2.36{\mu}m$ vs. $101.46{\pm}1.07{\mu}m$, respectively). Moreover, the diameters of the outer holes of the ZP were significantly (p < 0.05) larger in in vivo matured ($1.48{\pm}0.42{\mu}m$) than in vitro matured for 72 and immature oocytes ($1.10{\pm}0.16$ and $0.43{\pm}0.12{\mu}m$, respectively). Taken together, these data indicates that the ZP surface is related to oocyte maturity in canine.

• Animal Bioscience
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• v.34 no.4
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• pp.546-557
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• 2021

Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

• Journal of Marine Life Science
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• v.5 no.2
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• pp.58-63
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• 2020

The ascidian Halocynthia aurantium (sea peach), which belongs to the phylum Chordata, is thought to be a valuable organism of aquaculture like H. roretzi (sea pineapple), but its biological characteristics such as development and ecology are not well known. In this study, in order to obtain basic data for H. aurantium farming, the development processes of H. aurantium inhabiting the east coast of Gangwon-do were investigated and compared with those of H. roretzi, a related species. As a result, the morphology and developmental stages of the fertilized eggs, embryos and larvae of H. aurantium were very similar to those of H. roretzi. Fertilized eggs of H. aurantium took about 42.1 hours to hatch at 11℃, almost similar to 40.9 hours of H. roretzi. The time required for larvae to metamorphose into juveniles after hatching was very similar between the two species. The hatched larvae of the two species became juveniles with oral and atrial siphons after 23 days at 11℃. Both types of embryos developed slowly in seawater at low temperatures and rapidly developed at high temperatures. Fertilized eggs of H. aurantium hatched in an average of 62.3 hours at 9℃, 42.1 hours at 11℃, and 36.3 hours at 13℃, whereas those of H. roretzi hatched in an average of 60.4 hours, 40.9 hours, and 35.2 hours. Most of H. aurantium embryos did not develop normally above 15℃, so it is thought that attention is needed in the seed production processes.

• Journal of Dental Rehabilitation and Applied Science
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• v.40 no.1
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• pp.31-38
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• 2024

Dens in dente is a developmental anomaly resulting from infolding of the enamel organ into dental papilla prior to calcification of dental tissue. The pulpal tissue of the tooth can be vulnerable for bacterial invasion through direct exposure to the oral cavity or through defective enamel and dentin of the infolding part, thereby increasing the possibility of pulpal necrosis and subsequent apical periodontitis. Treatment planning of teeth with dens invaginatus may be difficult due to the complex root canal morphology. Therefore, thorough knowledge of anatomical variations of dens invaginatus is of great importance for proper treatment planning. The focus of this case report is on Oehler's type II and III dens invaginatus. The infolding of type III dens invaginatus extends beyond the crown and CEJ. Bacterial invasion through the infolding can easily cause inflammation of the pulpal and periradicular tissue. This case report presents endodontic treatment of type II and III dens invaginatus with the aid of CBCT.

• Development and Reproduction
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• v.28 no.3
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• pp.95-108
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• 2024

This study investigated the fin development and morphological characteristics according to larval growth in order to obtain information on behavioral characteristics and optimal stocking density during red seed grouper seed production. To examine the growth and fin development process of the larvae, we randomly sampled at 1, 3, 5, 7, 9, 10, 11, 13, 15, 17, 19, 20, 25, 30, 39, 45, 51, and 72 days after hatching. External morphology was observed and measured using an optical microscope. To observe skeletal development, larvae at 13, 20, 30, and 72 days after hatching were fixed in formalin and stained for cartilage and bone examination. At 9-10 DAH, red spotted grouper larvae (2.74±0.1 to 3.0±0.2 mm TL) exhibited a second dorsal fin spine and pelvic fin spine, which subsequently elongated. At 19-20 DAH, the larvae (5.7±0.1 to 6.1±0.1 mm TL) have the lengths of the second dorsal fin spine and pelvic fin spine average 34% and 31% to total length, respectively. From 30 to 72 DAH (12.6±0.4 to 56.0±0.2 mm TL), the length of the second dorsal fin spine and pelvic fin spine to total length decreased from 27% to 8% for the dorsal fin and 21% to 14% for the pelvic fin, respectively. At 30 DAH (12.6±0.4 mm TL), the larvae reached the complete count of fin rays in each fin. At 39 DAH (20.28±3.07 mm TL), the larvae had fin shapes similar to those of adults. At 13-30 DAH (4.2±0.1 to 12.6±0.9 mm TL), barbs and spinules were distributed along the ridges of the second dorsal and pelvic fin spines. However, at 72 DAH, these barbs and spinules were no longer observed on the fins. During the seed production process, red spotted grouper larvae tend to cluster in the morning, and during this time, entanglement of barbs and spinules on the second dorsal and pelvic fin spines can lead to mortality. Therefore, it is considered essential to focus on managing the behavioral patterns and appropriate rearing density of red spotted grouper larvae from the emergence of barbs and spinules on the second dorsal and pelvic fin spines until they regress and metamorphosis is completed.

• Journal of dental hygiene science
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• v.13 no.2
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• pp.203-212
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• 2013

The tooth morphology and qualitative mineral contents on enamel surface using energy dispersive X-ray spectroscopy, (EDX) were examined in the white-toothed shrew (genus Crocidura ) Crocidura lasiura and C. suaveolens and the red-toothed shrew (genus Sorex) Sorex caecutiens. In the case of C. lasiura and C. suaveolens, dental formula was found I 3/1 C1/1 P1/1 M3/3=28. The upper 1st and 2nd molars had an unequal W-shape formed by 5 cusps on the crown. The 3rd molar was found one-third the size of those of 1st and 2nd molars. The upper 1st incisor had two different sized hook-shapes and the lower 1st incisor was even. In the case of S. caecutiens, dental formula was found to be I3/1 C1/1 P3/1 M3/3=32. The upper 1st and 2nd molars had an equal W-shape on crown. The upper 3rd molar was half the size of those of the other molars. The upper 1st incisor possessed two similar sized hook-shapes and the lower 1st incisor had an uneven and serrated form. A comparison with the dental and cranial measurements revealed C. lasiura to be the largest of the three species (p<0.001) and C. suaveolens and S. caecutiens were similar in size (p>0.05). A qualitative analysis of mineral contents on enamel surface of the lower 1st incisor and lower 1st molar using EDX revealed C, O, P, Ca and Cu in all specimens and Pb was detected in several enamel specimens. No significant differences in the mineral contents (% weight) were observed among the three species (p>0.05). Fe was only detected on enamel surface of S. caecutiens with red pigmented teeth. Therefore, Fe is responsible for the red tip of the teeth. These results suggest that tooth morphological characteristics including the color of the tooth tip might be used as the key classifying species belonging to Crocidura and Sorex.

• Development and Reproduction
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• v.12 no.3
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• pp.221-229
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• 2008

For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.