• Journal of Embryo Transfer
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• v.33 no.4
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• pp.321-326
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• 2018

In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to $40{\times}10^6cells/mL$ was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and $2505.2{\times}10^6cells/mL$, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with $89.5{\pm}12.8$ and $91.4{\pm}7.9%$, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.235-243
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• 2000

Objective: Our present studies were conducted to examine more effective isolating method of preantral follicles from mouse ovaries. Methods: ICR mice (3-6 weeks old) were sacrificed through cervical dislocation and their ovaries were removed and put into watch glasses containing Hams F-10 supplemented with 10% fetal bovine serum (FBS). Preantral follicles were isolated by three different methods; 1) enzymatical method and 2) mincing method, and 3) scraping method. Enzymatical method was carried out as following. Ovaries were bisected with a pair of fine 30G needles. Bisected ovaries were incubated at $37^{\circ}C$ and 5% $CO_2$ incubator in 2-well dish containing Hams F-10 supplemented with collagenase 600 lU/ml and DNAse 20 lU/ml. After 20 min., follicles were isolated by repeated pipetting. Isolated preantral follicles were collected, and the remnant of tissues was placed in incubator and previous procedure was repeated. Mincing method was carried out with a pair of fine 30G needles attached to 1 ml syringes and minced ovary. Scraping method was carried out with a pair of fine 30G needles and scratched to surface of ovary. The differences between isolating methods were analyzed using Student's t-test and Chi-square. Results were considered statistically significant when ${\rho}$ value was less than 0.05. Results: In handling time, mincing or scraping method ($28{\pm}3.42$ min or $16{\pm}1.58$ min) were significantly (p<0.00001) shorter than enzymatical method ($72{\pm}1.69$ min), and scraping method was significantly (p<0.01) shorter than mincing method. Total number of isolated follicles was significantly (p<0.0001) higher in enzymatical method ($49.8{\pm}3.91$) than in mincing or scraping method ($25.3{\pm}2.33$ or $20.5{\pm}1.75$). Isolated follicles in ${\leq}$90${\mu}m$ were significantly (p<0.005) higher in enzymatical method ($15{\pm}1.71$) than in mincing or scraping method ($7.8{\pm}0.98$ or $8.1{\pm}1.31$). In 91~130 ${\mu}m$, isolated follicles were significantly (p<0.0005) higher in enzymatical method ($33{\pm}3.27$) than in mincing or scraping method ($16.3{\pm}1.82$ or $10.7{\pm}1.38$). In ${\geq}$ 131 ${\mu}m$, isolated follicles were not significantly differences between all groups. In equal sizes, the rate of isolated follicles in ${\leq}$ 90 ${\mu}m$ was highest in scraping method (39.6% vs. enzymatical method: 30.1%, p<0.05; mincing method: 30.9%, p=0.11719, NS). Rate of follicles in $91{\sim}130$ ${\mu}m$ was significantly (p<0.05) lower in scraping method (52.7%) than in enzymatical or mincing method (66.3% or 64.5%). Rate of follicles in ${\geq}$131 ${\mu}m$ was highest in scraping method (8.3% vs. enzymatical or scraping method: 3.6%, p<0.05 or 4.6%, p=0.19053, NS). Conclusions: This study suggests that scraping method is simple and useful for isolation of preantral follicles, because this method reduced handling time and recovered enough follicles. The recovered rate of isolated follicles in diameter of 91 ~ 130 ${\mu}m$ was highest in all methods.

• Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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• 1997.04a
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• pp.266-271
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• 1997

본 연구는 믹서기의 다지기칼날(Mincing Knife)이 구조적으로 가지고 있는 모멘트불평형(Couple Unbalance) 문제를 해결하기 위한 밸런싱방법을 제시한다. 유한요소법에 의하여 칼날에 작용하는 원심력, 정적 불평형량(Static Unbalance) 및 불평형각위치(Unbalance Angle)를 계산하고, 이를 바탕으로 반고리(Semiannular) 형상의 질량을 설계하여 모멘트 밸런싱을 수행하였다. 밸런싱을 수행한 후의 다지기칼날은 분해칼날(Cutting Knife)과 거의 비슷한 동적거동을 가지게 됨을 확인하였다.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.1
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• pp.91-95
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• 1992

Two experiments were conducted to evaluate the performance of broilers and layers fed locally produced fish waste meal (LFWM) in Western Samoa. LFWM, which was produced by mincing and oven drying of rejects from filleted fish, was included in both broiler and layer diets at the complete expense of imported meat and bone meal (MBM). The experiments were of a completely randomised design. In experiment 1, 120 day-old Strabro broiler chicks were raised to 49 days of ages. In experiment 2, 80 Shaver 288 pullets, 20 weeks of age, were housed in pairs in cages and raised up to 72 weeks of age. There were no differences (p > 0.05) in performance of broilers as measured in terms of final bodyweights, feed intake, feed efficiency and carcass yield between the two diets. Similarly, no differences (p > 0.05) existed in egg production, egg weight and feed required per kilogramme of eggs. It is concluded that LFWM is nutritionally as good as imported MBM and could be used to completely replace MBM in broiler and layer diets.

• Korean Journal of Animal Reproduction
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• v.17 no.3
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• pp.249-255
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• 1993

Canine follicular oocytes were used to establish a reliable system for maturation and fertilization in vitro. Ovaries were obtained from either slaughter house or hormone-primed bitches of mixed breeds. The oocytes were recovered by mincing the ovaries in M2＋BSA. Good quality of oocyte-cumulus complexes (OCCs) were selected and cultured in TCM 199 containing 15% fetal calf serum(FCS) for 24~56 h in an atmosphere of 5% CO2 at 39$^{\circ}C$. Maturation rate of follicular oocytes was >87% showing metaphase I. Unlike other domestic animals the cumulus expansion did not occur fully in canine OCCs although minimum expansion was found between the cumulus cells and corona radiata cells, the clear nuclear morphology was presented for the first time by rapid staining. The IVM system used in this study may be useful to obtain fully maturated metaphase I oocyte in dog.

• Korean Journal of Pharmacognosy
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• v.37 no.2 s.145
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• pp.97-102
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• 2006

Garlic(Allium sativum Linn) is widely used as a common condiment for a variety of foods and beverages. It has been well known that fresh garlic and garlic supplement of commercial preparations have various therapeutic properties including antimicrobial activity, antiplatelet aggregation, antihypertension, and cholesterol-lowering effects, which contribute to its increasing uses for an alternative medicine. Allicin(diallyl thiosulfinate), the major bioactive components of garlic, is formed by alliinase cleavage of the naturally occurring alliin upon crushing or mincing of garlic, and is the progenitor of a number of other products, such as diallyl disulfide. CYP3A4, heme-containing monooxygenase, is a key enzyme responsible for drug metabolism. Therefor, in the present study, we isolated and examined the compounds with CYP3A4-inhibiting activities from garlic. Among EtOAc extracts of garlic, we found that N-p-coumaroyltyramine and N-feruloyltyramine showed remarkable CYP3A4-inhibiting activities, compared to diallyl disulfide. Structures of the isolated active compounds were established by chemical and spectroscopic means.

• Reproductive and Developmental Biology
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• v.37 no.3
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• pp.143-148
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• 2013

Spermatogenesis is initiated from spermatogonial stem cells (SSCs) that has an ability of self-renewal and unipotency to generate differentiating germ cells. The objective of this study is to develop the simple method for derivation of SSCs using non-sorting of both spermatogonia and feeder cells. Simply uncapsulated mouse testes were treated with enzymes followed by surgical mincing, and single cells were cultured in stempro-$34^{TM}$ cell culture media at $37^{\circ}C$. After 5 days of culture, aciniform of SSC colony was observed, and showed a strong alkaline phosphatase activity. Molecular characterization of mouse SSCs showed that most of the mouse SSC markers such as integrin ${\alpha}6$ and ${\beta}1$, CD9 and Stra8. In addition, pluripotency embryonic stem cell (ESC) marker Oct4 were expressed, however Sox2 expression was lowered. Interestingly, expression of SSC markers such as Vasa, Dazl and PLZF were stronger than mouse ESC (mESC). This data suggest that generated mouse SSCs (mSSCs) in this study has at least similar biomarkers expression to mESC and mSSCs derived from other study. Immunocytochemistry using whole mSSC colony also confirmed that mSSCs generated from this study expressed SSC specific biomarkers such as c-kit, Thy1, Vasa and Dazl. In conclusion, mSSCs from 5 days old mouse testes were successfully established without sorting of spermatogonia, and this cells expressed both mESC and SSC specific biomarkers. This simple derivation method for mSSCs may facilitate the study of spermatogenesis.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.26-31
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• 2004

This work was undertaken in order to study the developmental competence of nuclear transfer cat embryo with fetal fibroblast and adult skin fibroblast as donor nuclei. Oocytes wererecovered by mincing the ovaries in Hepes-buffered TCM199 and selected the cumulus oocyte complexes (COCs) with compact cumulus cell mass and dark. Homogenous ooplasm were cultured for maturation in TCM199 + 10% fetal bovine serum (FBS) for 12 hours and used as a source of recipient cytoplast for exogenous somatic nuclei. In Experiment 1, we evaluated the effect donor cell types on the reconstruction and development of cloned embryos. Fusion, first cleavage and blastocyst developmental rate was not different between fetal fibroblast and adult skin cell (71.2 vs. 66.8; 71.0 vs. 57.6; 4.0 vs. 6.1 %, P<0.05). In Experiment 2, cloned embryos were surgically transferred into the oviducts of recipient queens. One of seven recipient queens was delivered naturally 2healthy cloned cats and 1 stillborn from fetal fibroblast cell of male origin after 65 days embryo transfer. One of three recipient queens was delivered naturally 1 healthy cloned cat from adult skin cell of female after 65 days embryo transfer. The cloned cats showed genotypes identical to the donor cell lines, indicating that adult somatic cells can be used for feline cloning.

• BMB Reports
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• v.56 no.4
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• pp.258-264
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• 2023

As a high-grade soft-tissue sarcoma (STS), undifferentiated pleomorphic sarcoma (UPS) is highly recurrent and malignant. UPS is categorized as a tumor of uncertain differentiation and has few options for treatment due to its lack of targetable genetic alterations. There are also few cell lines that provide a representative model for UPS, leading to a dearth of experimental research. Here, we established and characterized new cell lines derived from two recurrent UPS tissues. Cells were obtained from UPS tissues by mincing, followed by extraction or dissociation using enzymes and culture in a standard culture environment. Cells were maintained for several months without artificial treatment, and some cell clones were found to be tumorigenic in an immunodeficient mouse model. Interestingly, some cells formed tumors in vivo when injected after aggregation in a non-adherent culture system for 24 h. The tissues from in vivo study and tissues from patients shared common histological characteristics. Pathways related to the cell cycle, such as DNA replication, were enriched in both cell clones. Pathways related to cell-cell adhesion and cell-cell signaling were also enriched, suggesting a role of the mesenchymal-to-epithelial transition for tumorigenicity in vivo. These new UPS cell lines may facilitate research to identify therapeutic strategies for UPS.

• The Korean Journal of Physiology
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• v.2 no.2
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• pp.63-73
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• 1968

It is well known that mammalian alveolar membrane is covered with a very thin layer of surfactant film which characteristically reduces surface tension of alveolar membrane, and maintains alveolar stability. Since Clements in 1957 demonstrated that the surfactant is extractable by mincing the lung tissue in saline, various studies on the subject have been succeeded by many workers. However, the effect of radiation on the surfactant is not well clarified. Present study was attempted to observe the effect of x-irradiation on the activity of surfactant in rabbits. X-ray in dose of 300 r, 600 r or 900 r was irradiated to the chest of rabbits. The lung was removed from normal or irradiated rabbits sacrificed by arterial blood shedding, and lung-saline extract, adding 3 grams of lung tissue to 50 mili-liters of saline, was prepared by means of Vertis homogenizer. Tension-area diagram of lung extract was recorded automatically by a modified Langmuir-Wilhelmy balance with a synchronized recording system designed in this department. The surface tension of lung extract was measured at 1st, 2 nd, 3 rd, 7 th and 15 th post-radiation day in 300 r irradiated group, at 3 rd, 7 th and 15 th post-radiation day in 600 r irradiated group, 3 rd and 7 th post-radiation day in 900 r irradiated group respectively. For the histo-pathological study, lung tissue preparations were made in all irradiatiated groups on the day of experiment and in normal group. The results obtained are summarized as follows: 1. The minimal surface tension, maximal surface tension and stability index of normal rabbits lung extracts were 7.68 dynes/cm, 38.84 dynes/cm, and 1.39 respectively. 2. The activity of surfactant was depressed prominently by x-irradiation. However, increase in the dose of x-irradiation did not show any significant change in the degree of surfactant activity suppression. The most marked depression in surfactant was observed at the third post-radiation day in all irradiated groups. 3. Activity of surfactant depressed by x-irradiation showed a tendency of recovering to normal on 15 th post-radiation day. 4. The tendency of change in activity of surfactant following x-irradiation was somewhat correlative with histo-pathological changes. But the degree of depression of surfactant by x-irradiation did not correspond to the degree of histo-pathological changes, and recovery of lung tissue from radiation damage, tissue edema and congestion, seemed to be followed by restoration of surfactant activity. 5. The width of the tension-area diagram was measured at the surface area of 40% in lung extract of normal and x-irradiated rabbits. And it was found that the changes of the width corresponded well with that of minimum surface tension and of stability index in all normal and x-irradiated groups.