• Korean Journal of Animal Reproduction
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• v.16 no.1
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• pp.55-62
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• 1992

Bovine oocytes obtained from ovarian(2 to 5mm in diameter) of slaughtered cows were cultured in TCM 199 with 10~20% estrous-cow-serum(ECS) for 24~25hr at 39$^{\circ}C$ in 5% CO2-95% air. After culture, some oocytes were examined their maturation. The remainder were used to assess the fertilizability with frozen-thawed spermatozoa in a medium containing caffeine and heparin, and subsequent development in media with bovien cumulus cells(BCC) or bovine oviduct epithelial cells(BOEC). The results obtained were summarized as follows ; 1. The maturation rate of the oocytes in TCM199 with 15% ECS group(76.5%) was higher than that of 10% ECS(69.2%) or 20% ECS group(64.8%). 2. The proportions of the oocytes penetrated and the pronuclear oocytes in the presence of caffeine and heparin were 72.1%(62/86) and 93.5%(58/62), respectively. The rate of polyspermy in the fertilized oocytes was 8.1%. 3. When 73 oocytes recovered from fertilization drop were cultured in TC-199 medium with 10% fetal calf serum(FCS), 41 oocytes(56%) cleaved to 2-cell and further stages of embryos. Among these only one embryo developed upto morula stage. 4. The rate of the cleaved oocytes was higher in medium with BCC(80%:59/74) than BOEC(76%:58/76). However, the rate of developed morulae and blastocysts was higher in the medium with BOEC(40%;23/58) than with BCC(34;20/59).

• Korean Journal of Animal Reproduction
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• v.19 no.1
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• pp.1-7
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• 1995

This study was accomplished to illuminate factors of contamination of microbes in the culture medium and effect of antibiotics on prevention of contamination in the medium when bovine follicular oocyte was matured, fertilized and developed in vitro. 1. When washed or unwashed semen diluted with TCM 199 was incubated for 24∼72hr, contamination was come out. 2. When diluted semen with TCM 199 which has penicillin, streptomycin, gentamycin, kanamycin or nystatin was incubated for 24∼72hr, contamination was not come out only in kanamycin. 3. When imported semen which was diluted in TCM 199 with penicillin, streptomycin, gentamycin, kanamycin or nystatin was incubated for 24∼72hr, contamination was not come out in all treatments. 4. When semen which was diluted in BO, CZB, Ham's F10 or TCM 199 was incubated for 24∼72hr, kanamycin showed no contamination in all treatments, but gentamycin showed contamination in CZB, Ham's F10 and TCM 199. 5. When the semen diluted in BO was moved at 24hr after incubation into BO and incubated for 72hr, contamination was not come out, but when it was moved into the TCM 199 and incubated for 72hr, contamination was come out at 48 to 72hr of incubation. 6. When the semen diluted in BO, BO＋BSA or BO＋FBS containing gentamycin, kanamycin or nystatin was incubated for 24∼72hr, the diluted semen in BO or BO＋BSA showed no contamination in all antibiotics but the diluted semen in BO＋FBS showed no contamination only in kanamycin. 7.The Pseudomonas cepacia, Serratia liquefaciens, Klebsiella pneumaniae was respectively isolated in the semen of A, B, and C bull and the microbes are highly affected by amikacin, tobramycin and kanamycin. 8. When bovine folicular oocyte was in vitro matured, fertilized and developed in the simple medium with kanamycin, 26.6% was developed to over 32cell stage embryo.

• Journal of Acupuncture Research
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• v.18 no.3
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• pp.143-153
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• 2001

Objective : To investigate the relation to the organs, shu points and mu points. The labeled common locations of the spinal cord and brain were observed following injection of pseudorabies virus(PRV) into the the kidney, UB23 and GB25. Methods : After survival times of 96 hours following injection of PRV, The fifteen rats were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by PRV immunohistochemical staining method, and observed with light microscope. Results : In spinal cord, PRV labeled neurons projecting to the kidney, BL23 and GB25 were founded in cervical, thoracic, lumbar and sacral spinal segments. Dense labeled areas of cervical segments were overlap in lateral cervical n. and lamina III-V area. Thoracic segments were overlap in lateral spinal n., intermediolateral n. and lamina V-X areas. Lumbar segments were overlap in lamina I-V areas. Sacral segments were overlap in lamina IV, V and X areas. In brain, PRV labeled areas projecting to the kidney, UB23 and GB25 were overlap in the A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular n./rostroventrolaterai n., raphe obscurus n,, raphe pallidus n., raphe magnus n., gigantocellular reticular n., locus coeruleus, subcoeruleus n., A5 cell group and paraventricular hypothalamic n.. Conclusions : This results suggest that PRV labeled overlap areas of projecting to the kidney may be correlated to shu and mu points related to the kidney. These morphological results provide that organs-shu(transport) and mu(alarm) points interrelationship may be related to the central autonomic pathways.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.4
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• pp.365-370
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• 2006

Viral, bacterial and fungal infections can be transmitted via allografts such as bone, skin, cornea and cardiovascular tissues. Allogenic bone grafts have possibility of transmission of hepatitis C, human immunodeficiency virus (HIV-1), human T-Cell leukaemia virus (HTLV), tuberculosis and other bacterias. The tissue bank should have a policy for obtaining information from the patient's medical report as to whether the donor had risk factors for infectious diseases. Over the past several years, improvements in donor screening criteria, such as excluding potential donor with "high risk" for HIV-1 and hepatitis infection, and donor blood testing result in the reduction of transmission of these diseases. During tissue processing, many allografts are exposed to antibiotics, disinfectants and terminal sterilization such as irradiation, which further reduce or remove the risk of transmitting diseases. Because the effectiveness of some tissue grafts such as, fresh frozen osteochondral grafts, depends on cellular viability, not all can be subjected to sterilization and processing steps and, therefore, the risk of transmission of infectious disease remains. This article is review of the transmission of considering infectious disease in allogenic bone transplantation and the processing steps of reducing the risk. The risk of viral transmission in allografts can be reduced in several standards. The most important are donor-screening tests and the removal of blood and soft tissues by processing steps under the aseptic environment. In conclusion, final sterilizations including the irradiation, can be establish the safety of allografts.

• Archives of Plastic Surgery
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• v.42 no.2
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• pp.143-149
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• 2015

Background Adipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at $-20^{\circ}C$ by using thawing methods used in clinics. Methods The survival rates of adipocytes in the following thawing groups were measured: natural thawing at $25^{\circ}C$ for 15 minutes; natural thawing at $25^{\circ}C$ for 5 minutes, followed by rapid thawing at $37^{\circ}C$ in a water bath for 5 minutes; and rapid thawing at $37^{\circ}C$ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes. Results In the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05). The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant. Conclusions It was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at $37^{\circ}C$. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

• Journal of Chest Surgery
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• v.40 no.11
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• pp.786-788
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• 2007

Primary malignant fibrous histiocytoma of the lung is extremely rare, A 12-year-old child was admitted to the hospital due to an incidentally detected lung mass. A $2.5{\times}2.5 cm$ sized mass located in the right lower lobe was seen on the chest X-ray, the chest computed tomogram and the positron emission tomogram. We resected the mass through a right lateral thoracotomy and the mass was revealed to be a malignant spindle cell neoplasm on the frozen section diagnosis. So, we performed lobectomy of the right lower lobe with systemic dissection of the mediastinal lymph nodes; the final histopathological diagnosis of the mass was malignant fibrous histiocytoma. The patient was discharged on postoperative day 7 and adjuvant chemotherapy was not applied.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.21 no.2
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• pp.97-104
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• 1988

Condensed phosphates have been used in various meat products to enhance the water holding capacity, to improve texture and to prevent the development of off flavors and off odors. This study was intended to observe the effects of poly - and pyrophosphate on the growth of sanitary indicative bacteria and food poisoning bacteria. The bacteriostatic effect of poly - and pyrophosphates against Gram positive bacteria was much stronger than that of against Gram negative organisms. The effective inhibitory concentration of sodium polyphosphate on the growth of bacteria was varied by species such as $0.3\%$ to Staphylococcus aureus, $0.9\%$ to Salmonella, and more than $1.0\%$ to Escherichia coli in nutrient broth. When Staphylococcus aureus suspension was treated with $0.5\%$ sodium polyphosphate at $35^{\circ}C$ for 1 hour, the release of UV-absorbing substances from the organism was confirmed. However no significant effect was observed in Escherichia coli under the similar condition. When alaska pollack fillets were dipped in $ 3.5\%$ sodium polyphospahate at $2^{\circ}C$ for 1 min. prior to freezing, the viable cell count and coliform MPN's of the frozen product were decreased with the range of 30 to $50\%$ in comparison with those of control.

• The Journal of Korean Medicine
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• v.22 no.2
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• pp.31-40
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• 2001

This experimental studies was to investigate location of labeled neurons in CNS following injection of pseudorabies virus(PRV), Bartha strain, into the uterus and Sanyinjiao(Sp6) of rats. After survival times of 4-5 days following the injection of PRV, the rats were perfused, and their brain and spinal cord were frozen sectioned($30\mu\textrm{m}$). These sections were stained by PRV immunohistochemical staining methods, and observed with light microscope. The results were as follows: 1. In the spinal cord, overlap areas of PRV labeled neurons projecting to uterus and Sp6 were observed in lamina VII, IX and X areas of cervical segments. In thoracic segments, overlap areas were observed in lamina IV, VII, X and intermediolateral n.. In lumbar segments, overlap area of PRV labeled neurons were observed in lamina I, V-VII, IX, X and intermediolateral n.. In sacral segments, overlap areas of PRY labeled neurons were observed in lamina N, V, VII, X and sacral parasympathetic n.. 2. In the brain, overlap areas of PR V labeled neurons projecting to the uterus and Sp6 were observed in lateral paragigantocellular n., rostroventrolateral reticular n., raphe obscurus n., raphe pallidus n., raphe magnus n., locus coeruleus n., Barrington's n., A5 cell group, central gray n., paraventricular hypothalamic n. and arcuate n. This results suggest that overlap areas of PRV labeled neurons of the spinal cord projecting to the uterus and Sp6 might be the first-order neurons related to the viscera-somatic sensory and sympathetic preganglionic neurons. PRV labeled neurons of the brain may be the second and third-order neurons response to the movement of smooth muscle of uterus. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitoring the internal environment. These overlap areas of spinal cord and brainmay be related to autonomic centers related to regulation of uterus.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5731-5734
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• 2013

Background: The object of this study was to examine whether a new protocol including step-sectioning and immunohistochemistry (IHC) staining of axillary sentinel nodes (SN) would lead to detection of more metastases in patients with breast cancer. Materials and Methods: Sixty-nine tumor free sentinel lymph nodes were examined. Step frozen sectioning was performed on formalin fixed SN and stained both by hematoxylin and eosin (H and E) and cytokeratin markers using IHC. Any tumoral cell in IHC stained slides were considered as a positive result. Metastases up to 0.2 mm were considered as isolated tumor cells and 0.2 up to 2 mm as micrometastasis. Results: Mean age of the patients was $48.7{\pm}12.2$ years. Step sectioning of the SN revealed 11 involved by metastasis which was statistically significant (p<0.001). Furthermore, 15 (21.7%) of the patients revealed positive results in IHC staining for pan-CK marker and this was also statistically significant (p=0.001). Ten patients had tumoral involvement in lymph nodes harvested from axillary dissection and 4 out of 15 lymph nodes with positive result for CK marker were isolated tumor cells. However, 4 of 10 patients with tumor positive lymph nodes in axillary dissection were negative for CK marker and in contrast 6 of the pan-CK positive SN were in patients with tumor-free axillary lymph nodes. Conclusions: Both IHC and step sectioning improve the detection rate of metastases. Considering the similar power of these two methods, we recommend using either IHC staining or step sectioning for better evaluation of harvested SNs.

• Journal of Embryo Transfer
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• v.32 no.4
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• pp.337-342
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• 2017

The present study was aimed to determine the effects of green tea extract (GTE) and beta-mercaptoethanol (${\beta}-ME$) supplementation in boar sperm freezing extender on in vitro fertilization (IVF) and reactive oxygen species (ROS) and glutathione (GSH) levels of presumptive zygotes (PZs). Experimental groups were allocated into lactose egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/l in LEY) and ${\beta}-ME$ ($50{\mu}M$ in LEY). In freezing, spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM. The final sperm concentration was $1{\times}10^8/ml$. Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. For IVF, oocytes were matured in NCSU-23 medium and co-cultured with spermatozoa following thawing at $37^{\circ}C$ for 25 sec. At 12 h following IVF, IVF parameters (sperm penetration and monospermy) were evaluated. In addition, GSH and ROS levels of PZs were determined by Cell Tracker Blue CMF2HC and DCHFDA, respectively. IVF parameters did not show any significant difference among the experimental groups. GSH and ROS levels of PZs were not significantly different between groups. In conclusion, antioxidant supplementation in boar sperm freezing could not influence IVF parameters, ROS and GSH levels of PZs.