• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.301-309
• /
• 1997

The objective of this study was to determine the microtubule assembly and chromatin configuration during the first cell cycle in bovine oocytes following injection of spermatozoon, sperm head and tail. The microtubule and chromatin configuration was imaged with fluorescent labeled monoclonal ${\alpha}$-tubulin antibody and propidium iodide under laser scanning confocal microscope. Microtubule and chromatin dynamics in bovine oocytes following intracytoplasmic sperm injection (ICSI) were not different from those observed during in vitro fertilization (IVF). Following ICSI, the microtubular aster was observed around sperm midpiece. During pronuclear formation, the sperm aster was enlarged and seen around male and female pronuclei. At mitotic metaphase, the microtubular spindle assemble astral poles and chromosomes were aligned on the spindle equator. At mitosis, asters were concentrated to each spindle pole and they filled the cytoplasm. After injection of the isolated sperm head, the microtubular aster was not seen around sperm head in any cases (0/18). Instead, microtubules were organized from the cytoplasm, which filled the whole cytoplasm during pronuclear apposition. These microtubules seem to move male and female pronuclei. These results suggest that isolated sperm head can develop into normal pronucleus in mature bovine oocytes, and competent to participate syngamy with the ootid chromatin. The functional microtubules following isolated sperm head injection in bovine oocytes appeared to be organized solely from maternal stores.

• The Korean Journal of Mycology
• /
• v.48 no.2
• /
• pp.151-160
• /
• 2020

The goal of this study was to investigate the diversity present among wild yeasts obtained from soils of spice fields and from mountain soils, and to further, characterize previously unrecorded novel wild yeast strains. In total, 36 strains from 17 different species of wild yeasts were isolated from 35 soil samples obtained from garlic fields of Geumsan, Chungcheongnam-do, Korea. Among these, six yeast strains of Trichosporon moniliiforme, and four strains each of Papiliotrema flavescens and Candida melibiosica species were isolated. Additionally, 22 strains of 18 different species of wild yeasts were isolated from 32 soil samples collected from the ballonflower and ginger fields of Geumsan, Korea. Finally, 46 strains of wild yeasts were isolated from 35 soil samples obtained from Mt. Daedun in Geumsan, Korea. Among the total of 106 isolated wild yeast strains, 10 strains, including Debaryomyces vindobonensis GHY31-3 represented novel yeast strains which were previously unrecorded. All the 10 previously unrecorded yeasts were oval or global in shape, and five strains, including Filobasidium stepposum SFG1-4 formed ascospores. Three strains, including Pseudozyma alboarmeniaca CD 23-5 grew well in vitamin-free medium. Cell-free extract obtained from Filobasidium magnum SFG1-3 indicated 28.6% of xanthine oxidase inhibitory activity.

• Korean Journal of Medicinal Crop Science
• /
• v.2 no.1
• /
• pp.81-85
• /
• 1994

This experiment was carried out to investigate the fertilization and the size of mature female and male gametophytic parts of Pulsatilla koreana after artificial pollination. The size of pollen is $26.5{\mu}m$ at the time of anther dehiscence, that is, about $3{\sim}4$ days after pollination. Synergid nucleus, egg nucleus, and polar nucleus are 10.0, 15.0, and $32.5{\mu}m$ respectively at the time of completing egg apparatus formation, that is, about 2 days after pollination. Poller tubes germinate on stigma about 10 hours, passing lower part of style about 30 hours, penetrating into micropyle about 35 hours after pollination. Sperm nucleus penetrates into polar nucleus about 40 hours and egg cell about 48 hours after pollination. But, there seems to be different among the individuals. Multinuclei and multinucleoli are formed in egg cell, synergid, and polar nucleus about the time of fertilization. Proembryo is formed about 4 days, being changed to large globular form about $6{\sim}8$ days after pollination. Endosperm nuclei divide into free nuclei after fertilization and change to cotyledon in gymnosperm. There seems to be same phenomena in Pulsatilla koreana.

• The Korean Journal of Mycology
• /
• v.38 no.2
• /
• pp.160-166
• /
• 2010

A homothallic filamentous fungus Aspergillus nidulans has been used as the a model organism for studying growth and development for eukaryotic system. Various studies about specific transcription factors have been performed for elucidating the molecular mechanisms of growth, asexual and sexual developmental processes. Among them, the fkhE gene (AN2025.3) is located in chromosome VII and contains an ORF encoding 718 amino acid polypeptide intervening with two short introns. The cDNA sequencing revealed that at least four types of alternative splicing events were occurred when the fkhE gene was transcribed. The putative FkhE polypeptide contains a conserved forkhead domain and a bipartite nuclear localization signal at it's N-terminus and C-terminus, respectively. Deletion of fkhE resulted in impaired conidiophore formation in a solid medium. However, the sexual developmental process or cleistothecia formation was normal. Furthermore, fkhE deletion mutant produced conidiophores and conidia under the submerged culture, indicating that the fkhE gene is involved in asexual developmental process similar to the fkhF gene.

• Radiation Oncology Journal
• /
• v.4 no.2
• /
• pp.99-105
• /
• 1986

Optimal balance between control probability and risk of complication is emphasized even in present time, Although certain incidence of intestinal injury is accepted as an inevitable consequence after abdominopelvic irradiation, these complications still remain as problems. 60 mice were irradiated with 250 kVp orthovoltage x-ray machine and $200rad{\times}5/wk$ regimen. Histpathologic findings of colorectum and the relationship with occult blood test were analyzed and possible tolerable dose which would be safe from permanent complication was also estimated. Followings are the results: Mild mucosal and submucosal edema were observed in 1,000 rad irradiated group. Congestion of small vessels was prominent in 2,000 rad irradiated group and infiltration of inflammatory cells was observed in 3,000 rad irradiated group. Denuded mucosa was observed in 3,000 rad irradiated group. Occult blood test is not a proper indicator for rectal denuding or rectal ulcer, but our results suggest the possibility of using this as a relative scale of intestinal damage. Mitotic figures of crypt cells were observed even in 5,000 rad irradiated group, these suggest that the repair capacity of crypt cells are still functioning.

• Korean Journal of Food Science and Technology
• /
• v.32 no.6
• /
• pp.1389-1395
• /
• 2000

Cervical cancer has been one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papillomaviruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, Rhus, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Rhus inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, Caski) and HaCaT keratinocytes in a dose response manner. In vitro binding assay and ELISA showed that Rhus inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Rhus inhibited the in vitro binding of E7 and Rb which essential tumor suppressor for the control of cell cycle. The level of mRNA for E6 was also decreased by Rhus while that of E7 mRNA was not changed. Our data suggested that Rhus inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas by HPV.

• Journal of Plant Biotechnology
• /
• v.46 no.3
• /
• pp.180-188
• /
• 2019

Auxin plays a crucial regulatory role in plant growth and development processes. Three major classes of auxin-responsive transcription factors controlled by the Auxin/indole-3-acetic acid (Aux/IAA), Gretchen Hagen 3 (GH3), and small auxin up RNA (SAUR) genes regulate auxin signaling. Aux/IAA, in particular, encodes short-lived nuclear proteins that accumulate rapidly in response to auxin signaling. In this study, we isolated a PagAux/IAA1 gene from poplar (Populus alba ${\times}$ P. glandulosa) and investigated its expression characteristics. The PagAux/IAA1 cDNA codes for putative 200 amino acids polypeptide containing four conserved domains and two nuclear localization signals (NLSs). Utilizing Southern blot analysis, we confirmed that a single copy of the PagAux/IAA1 gene was present in the poplar genome. The expression of this gene is specific to leaves and flowers of the poplar. PagAux/IAA1 expressed in the early exponential growth phase of cell-cultured in suspension. PagAux/IAA1 expression level reduced in drought and salt stress conditions, and the presence of plant hormones such as abscisic acid. However, expression enhanced in cold stress, cambial cell division, and presence of plant hormones such as gibberellic acid and jasmonic acid. Thus, these results suggest that PagAux/IAA1 participates in cold stress response as well as developmental processes in the poplar.

• Korean Journal of Animal Reproduction
• /
• v.24 no.4
• /
• pp.375-383
• /
• 2000

The nuclear maturation and developmental competence of immature, oocytes collected from donors at various morphology of corpus luteum (CL) and fertilized in vitro was investigated by comparing the meiotic activity and the yields of embryos. Ovaries were divided and classified into 4 groups as the following criteria : Group 1 ; ovaries showed evidence of recent ovulation (corpus hemorragicum). Group 2 ; apex of CL was red or brown. Vasculization was limited to periphery of CL. Group 3 ; apex of CL was orange or tan. Vasculization was covered over apex of CL. Group 4 ; CL was light yellow to white and firm in texture and the vascular network on the surface of CL had disappeared. Modified TCM 199 was used for maturation in vitro of immature oocytes and development was induced by using TLP-PVA as a basic medium. When oocytes collected from each group of donors had been matured for 4, 14, and 24 hours in vitro, the proportion of oocytes reaching metaphase I and metaphase II were not different among oocytes from 4 group of ovaries. Mature metaphase II stage of oocytes in each group was first observed at 14 hours, whereas completion of maturation of. oocytes in each group was at 24 hours. Luteal morphology of ovaries had little effect on the proportion of embryos reached 2 cells and 8 cell stage. However, the proportion of embryos cleaved to morula and blastocyst stage was significantly higher in the oocytes obtained from group 1 and 3 than in the oocytes from group 2 and 4 (p＜0.05). This data suggest that reproductive status of the donor significantly influence the yield of in vitro embryos.

• Journal of Plant Biotechnology
• /
• v.34 no.4
• /
• pp.331-338
• /
• 2007

One-node stem pieces ca. 1 cm in length containing a axillary bud and a fully expanded leaf were obtained from it in vitro plants of potato (Solanum tuberosum L.). Leaves were removed and the nodes were cultured on the MS medium to investigate the effects of temperature, day length, sucrose, and CCC in microtuber formation and development. The fresh weight of microtubers after 80 days increased significantly at 8% sucrose and $20^{\circ}C$ compared with $28^{\circ}C$. The tuberization and development were reduced at $28^{\circ}C$ except short-day treatment of 8 hours at 8% sucrose. The fresh weight and diameter were increased on the culture medium added CCC 500 mg/L. The potato tuberization was promoted under short daylength, and it showed great effect by treatment with the CCC. Though the tuberization was promoted at low temperature of $20^{\circ}C$ in a histologic change of an axillary bud part cell of a potato, the cells were able to observe the swelling growth. Swelling growth of tissue was stimulated in the darkness and was more remarkable by addition of CCC. In particular, in the visual ratio of cell division for each position in the tissue, the cortex part showed larger ratio of cell expansion than that of the pith part. The effect of CCC was identified at 8% sucrose in the darkness. The effect of CCC was not showed in sucrose 3% under long daylength of 16 hours. As a result, the fact of a substance with AGPase important for starch composition was certified by the result with the inclose of AGPase activity on high concentration of sucrose, CCC, and dark treatment by which tuber formation and development are promoted.

• Applied Microscopy
• /
• v.40 no.2
• /
• pp.73-80
• /
• 2010

Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.