• Title/Summary/Keyword: fixative

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Cytoskeletal Patterns, In Vitro Maturation and Parthenogenetic Development of Rabbit GV Oocytes

  • Ju, J.C.;Chen, T.H.;Tseng, J.K.;Tsay, C.;Yeh, S.P.;Chou, P.C.;Chen, C.H.;Liu, C.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.12
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    • pp.1695-1701
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    • 2002
  • The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.

Acrosome staining with Coomassie brilliant blue G or R on the horse spermatozoa (Coomassie brilliant blue G 및 R을 이용한 말 정액 첨체 염색 기법)

  • Kim, Sung Woo;Shin, Sang Min;Yu, Yeonhee;Lee, Jae-Yeong;Kim, Chan-Lan;Ko, Yeoung-Gyu
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.9
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    • pp.57-63
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    • 2020
  • To develop simple acrosome staining of horse spermatozoa, this study tested the binding properties of Coomassie brilliant blue G or R on the sperm smears after 3.7% paraformaldehyde (PF) or 35% methanol (MT) fixation. After being fixed with PF and stained with 0.05, 0.1, or 0.2 % of CBB G or R for 2 min, horse spermatozoa were examined for their intact acrosome status. The intact acrosome of fresh horse spermatozoa were 62.6% and 61.5% with 0.05% of the G and R CBB solution, but 80.2 and 79.7% with G type and 78.1 and 76.0% with R type. On the other hand, when MT was used for fixation, the acrosome reacting sperm ratio was 3.5%, but was 9.0% in the case of PF. These results show that the intact acrosome of horse sperm could be judged using a 0.1~0.2% CBB G or R staining technique. PF would be an essential fixative for examining acrosome reacting horse spermatozoa. This method could be used to identify sperm with a damaged acrosome during low-temperature storage or cryopreservation for artificial insemination of horses.

Ultrastructural Study on the Cerebellar Purkinje Cell of the Head-Irradiated Rat (과량의 방사선 국소조사가 흰쥐 소뇌 Purkinje세포의 미세구조에 미치는 영향)

  • Ahn, E-Tay;Yoon, Kyoo-Tae;Yang, Nam-Gil;Ko, Jeong-Sik;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.48-62
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    • 1994
  • The acute irradiation effect on rat Purkinje cell was carried out. Anesthetized rats, weighing 200-250g each, were exposed their heads to the linear accelerator (ML-4MV) with the doses of 3,000 rads or 6,000 rads respectively. Irradiated rats were sacrificed by perfusion fixation under anesthesia, six hours, two days and six days following the irradiations. Rats were perfused with the fixative of 1% glutaraldehyde-1% paraformaldehyde solution (pH 7.4). Small pieces of cerebellar cortices were taken out. Tissue blocks were washed out, and were refixed in the 2% osmium tetroxide solution. After dehydration, tissues were embedded in the araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution, were examined with an electron microscope. The results observed were as follow; 1. Many dark Purkinje cells exhibited most severe cellular alterations on 6 hours. But after the 2 or 6 days, the cells exhibited only some alterations of cytoplasmic organelles. 2. Many granular and agranular endoplasmic reticula exhibited the fusion of cisterns. These reticular alterations were most severe on 6 hours following irradiation. But the alterations were hardly found on 6 days. 3. In the Golgi region, alterations including the adhesion of lamelliform cisterns, enlarged saccules, and increased number of vesicles, etc, were seen on 6 hours. But the Golgi complexes were almost recovered on 6 days. 4. Lysosomes were abundant on 6 hours or 2 days, but some residual bodies were found on 6 days. 5. Mitochondrial changes were also most severe at on hours, and they were recovered thereafter. From the results, it was concluded that the cerebellar Purkinje cells reacted to the high doses of irradiation by hyperactive protein synthesis, autolytic activities and energy metabolism. The reaction was most active in the early stage. It implies that motor-control function of Purkinje cells are severely disturbed in the early stage of irradiation.

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Morphometric study of the Leydig cell in Korean native cattle (Bos taurus coreae) (한우 고환 간질세포의 형태계측학적 연구)

  • Tae, Hyun-Jin;Ahn, Dong-Choon;Lee, Young-Deok;Na, Sung-Jin;Park, Byung-Yong;Kim, In-Shik
    • Korean Journal of Veterinary Service
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    • v.39 no.3
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    • pp.141-149
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    • 2016
  • Changes in the Leydig cell from pre-puberty to adulthood were studied in Korean native cattle. Eight groups of male cattle aged 14, 17, 20, 25, 30, 35, 40 and 104 weeks (n=7 cattle per group) after birth were used. The purpose of this study was to obtain quantitative information on the Leydig cell of the Korean native cattle during postnatal development. Testes of cattle were fixed by perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ section stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. The average diameter of seminiferous tubules gradually increased with age from 14 ($75.56{\mu}m$) to 104 weeks ($298.9{\mu}m$). The volume density of the seminiferous tubules increased with age from 54.2% at week 14 to 76.9% at week 104. The volume density of the interstitium represents 45.52% at week 14. This proportion progressively diminishes during development to reach a value of 23.14% at week 104. The volume density of Leydig cells decreased almost linearly from 14 (20.71%) to 104 weeks (5.28%). The absolute volume of Leydig cells per testis increased significantly from 14 to 104 weeks. The number of Leydig per testis have almost linearly increased from 14 to 104 weeks. The average volume of a Leydig cell reached maximum size by 104 weeks ($2553{\mu}m^3$). These data suggested development of Leydig cell can be classified as the fetal and immature adult Leydig cells (14~35 weeks), and the adult Leydig cells (40~104 weeks).

A Study on the Weathering Resistance of Fixatives Used on Conservation of Painting Layer of Korea Wooden Painting (한국 판벽화 채색층 보존처리 고착제의 내후성 연구)

  • Lee, Seul;Han, Kyeong Soon
    • Journal of Conservation Science
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    • v.34 no.5
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    • pp.397-405
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    • 2018
  • Currently, there are very few studies on Korean wall paintings. Therefore, this study discusses the current conditions of wooden paintings and the characteristics of the adhesive agent in the painting layer separation. Korean land pine was chosen as the support, while white oysters shells, orpiment, red ocher, Noerok, and azurite were used as pigments. With four adhesive agents, including animal glue, Gelidium, methyl cellulose, and PVAc (caparol binder), a comparative experiment was conducted, by dividing them into two concentrations, of 0.5% and 1.5%. The temperature, humidity, and ultraviolet rays, which are contributing environmental factors in cultural assets after fixing, were artificially investigated. After deterioration, observed color difference, fixing, and the surface. Results showed that the animal glue strongly fixing all the colored layers compared to the other adhesives; however, azurite had a partial change when used outdoors. With Gelidium, which functioned similar to animal glue, the azurite was affected by the ultraviolet rays; nevertheless, despite the variations in temperature and humidity, it had the best gripping force compared to the other adhesive agents. Methyl cellulose was glossy at a high concentration, and was relatively strong against rapid changes in temperature and humidity. PVAc significantly reduced the binding force, compared to other adhesive agents.

Concept Analysis of the Role in Nursing Profession (간호전문직에서의 역할의 개념분석)

  • Kang, Sung-Rye;Park, Jung-Ho
    • Journal of Korean Academy of Nursing Administration
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    • v.8 no.3
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    • pp.431-439
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    • 2002
  • Purpose : Nursing profession has been rapidly changed due to the development of bioscience and the change of medical service environment. Nursing profession that has been focused on only care giver's role like mother's in tradition, is required careful reflection new nursing role to cope properly to the change of medical service environment. In nursing profession, the concept of the role was used without an accurate definition, as a results, the concept of the role was mixed with the concept of the "activity", "job" and "function". Therefore, concept analysis of the role in nursing profession is needed to remove conceptual ambiguous and make confirm the true meaning of the role. Concept analysis is a strategy that examine the characteristics of a concept. It is a formal linguistic exercise to determine those defining attributes. The basic purpose of concept analysis is to clarify ambiguous concept in theory, and propose operational definition which reflects the theoretical base of the concept. Method : In this study, the concept of the role in nursing profession which was analyzed based on the steps of concept analysis by Walker & Avant(1988). Results : The attributes of the concept of the role in nursing profession are : 1. Basic factors to construct social structure. 2. Expected attitudes, values, behavior patterns according to the social status and position in social structure. 3. Perceptive attitudes, values, behavior patterns in order to social interaction. 4. Acquiring through continuous communication and learning. 5. Not fixative but changeable. The antecedents of the role in nursing profession are : 1. The social status and position which is possessed by a person. 2. Expected attitudes, values, behavior patterns according to the social status and position in social structure. 3. Acquiring through continuous interaction, communication, and learning process. And the consequences of the role in nursing profession are : 1. The role maintenance and the role expansion as a positive consequences. 2. The role conflict and the role loss as a negative consequences. Conclusion : Through this concept analysis, the concept of the role in nursing profession is defined as As basic factors to construct social structure, person's attitudes, values, behavior patterns which are expected from other social members according to person's social status and position, and perceived and acquired through continuous interaction, communication, and leaning process.

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Some Observations on the Organelles Participating in the Biliary Excretion in the Hepatocyte of the Biligrafin Injected Mouse (Biligrafin 투여 마우스 간세포의 미세구조적 및 세포화학적 연구)

  • Kim, Hyang;Shin, Young-Chul
    • Applied Microscopy
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    • v.23 no.2
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    • pp.53-77
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    • 1993
  • In this study, an attempt was made to investigate the probable organelles participating in the secretion of biligrafin. The animals (ICR male mice, 25-30gm) were divided into normal control and 6 biligrafin injected groups to which 30% biligrafin (0.006ml/gm b.w.) were injected at 10, 20, 40, 80, 160 and 320 min prior to the sampling. The mice of each group were perfused through the heart with ice-cold 2.5% glutaraldehyde buffered with 0.1M Na-cacodylate (pH. 7.4) under the Na-pentobarbital (Nembtal 0.0015mg/gm b.w.) anesthesia and liver tissues were taken from each group. Some specimens were immersed 1 hr in the same solution used in the perfusion. After an overnight rinse in 0.1M Na-cacodylate buffer containing 10% DMSO and 7.6% sucrose, $75{\mu}m$ fronzen sections were made for cytochemical study. The sections were incubated in thiamin pyrophosphatase (TPPase) and inosine diphosphatase (ID Pase) media for 70 min at $37^{\circ}C$ respectively and acid phosphatase (AcPase) medium for 40 min at $37^{\circ}C$. They were postfixed in 1 % $OsO_4$ for 1 hr. The other specimens were immersed for 8 hrs in the fixative consisting of 2.5% glutaraldehyde and 3.0% paraformaldehyde buffered with Na-cacodylate (pH. 7.4). All of the osmificated specimens were processed for electron microscopy. In both normal and biligrafin injected groups, endoplasmic reticulum (ER), vacuoles, Golgi apparatus and lysosomes were seen in the vicinity of bile canaliculus. In the biligrafin injected groups, however, the Golgi apparatus appeared to be decreased and ER and vacuoles were dilated and increased. The rough endoplasmic reticulum (RER) having a few attached ribosomes appeared to be the round saccule, especially at 20 min after biligrafin injection. Smooth endoplasmic reticulum (SER) seemed to be formed by the detachment of ribosomes at the cisternal end of RER. The cistern of SER showed saccules which probably budded off to form the vacuole. The vacuoles were devoid of visible centents. This finding seemed to be in agreement with the biochemical property of the bile constituents. The fusion between the vacuoles and bile canaliculus were frequently seen in the groups injected with biligrafin. The lysosome did not show any changes in the biligrafin injected groups. Accumulation of some material and lipid droplets were seen at the 40 and 80 min after biligrafin injection, especially at the latter. At 160 and 320 min after biligrafin injections, however, they were decreased successively while the RER stack, free ribosomes and polysomes were increased. Although the reactive products of TPPase and IDPase were observed in the ER saccules and vesicles of the normal control and biligrafin injected groups, the fusion between the bile canaliculus and saccules or vesicles could easily be seen in the latter. The AcPase activity, however, was observed in the cistern at the maturing face of Golgi apparatus and lysosomes in both normal and biligrafin groups. The results suggest that the biligrafin is excreted via the vesicles, vacuoles or sacoules probably derived from the SER without the participation of Golgi apparatus and lysosomes, and the excess amount of material is stored as inclusions during the repairing of the organelles being overactive.

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Current Status and Perspectives of Quality Improvement in Sesame (참깨 품질 연구의 현황과 문제점 및 전망)

  • Lee, Bong-Ho;Lee, Jung-Il;Park, Rae-Kyeong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.33 no.s01
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    • pp.86-97
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    • 1988
  • Sesame(Sesamum indicum L.) is probably the most ancient oilseed crop known in the world. The seed of sesame is used in a variety of ways as food. The whole seed may be eaten raw, either roasted or parched, or fed to birds and stock. Sesame oil is used as a salad or cooking oil, in shortening, margarine and in the manufacture of soap. Minor uses are as a fixative in the perfume industry and formerly as a carrier for fatsoluble substances in pharmaceuticals such as penicillin. One of the minor constituents of sesame oil, sesamin, is used for its synergistic effect in pyrethrin insecticides, in addition of a small quantity of this substance markedly increases the effectiveness of fly sprays. The meal remaining after oil extraction can be used as and animal feed-stuff or as manure. In general sesame meal is considered to be equal to cottonseed or soybean meal as a protein supplement for livestock and poultry. It is especially high in certain amino-acids such as methionine, which is low in soybean meal, and thus can be combined with it or similar meal to form a more balanced ration. An attempt to summarize the literature review on quality improvement of sesame was made to discuss the accomplishments of the past and perspectives in the future. The reviews on quality improvement of sesame were mainly discussed in connection with the cultural practices and genetic informations in current status. The emphasis focussed on environmental variation of quality in cultural practices, such as harvest time, variety by location, climatic condition, fertilizer application, and growth regulator treatment. On the genetic variation of quality, it was discussed on variety background, mutation breeding, correlations, and inheritance of quality related characteristics. It also was discussed on relationship between quality and plant traits, storage condition or period, and seed coat color. Moreover, current research status were reviewed on some minor elements such as sesamin, oxalic acid, and trypsin inhibitor. As a results of the review, the lack of an effort to quality improvement in each utilization area was indicated as a problem area. More active efforts for the improvement of quality were also insufficient to incorporate the available genes for quality in breeding method or collection and analysis of breeding materials. Therefore, researches in the future would be recommended to emphasize on these problem areas.

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Ultrastructure of the Granular Glands in the Amphibian Skin (양서루 피부 과립선의 미세구조)

  • Kim, Han-Hwa;Noh, Yong-Tai;Chung, Young-Wha;Chi, Young-Duk
    • The Korean Journal of Zoology
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    • v.22 no.3
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    • pp.103-114
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    • 1979
  • The authors observed the ultrastructure of the granular glands in the amphibian skin with an electron microscope. The specimens from the experimental animals (Bombina orientalis, Bufo bufo gargarizans, Rana nigromaculata and Rana rugosa) were fixed in 2.5% glutaraldehyde-paraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with a LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follws: 1. The granular gland in the amphibian skin consisted of the glandular epithelial and the myoepithelial cells. 2. The epithelial cells of the granular gland in the amphibian skin consisted of the dark cells but the light cells were also observed in that of Bombina orientalis. 3. The granular glands of the amphibian skin were in holocrine fashion. 4. The nuclei of the epithelial cells of the amphibian cutaneous granular glands were round or oval and showed small and large inforldings of nuclear envelope. Heterochromatins were mainly distributed near the nuclear envelope. Mitochondria were mainly distributed in the perinuclear portion and rough-surfaced endoplasmic reticulums were developed in the cytoplasm but smooth-surfaced endoplasmic reticulums were not well developed. 5. Secretory granules were round or oval and electron-dense and less electron-dense granules were observed. 6. The authors infer that the differences in electron density of the secretory granules in the granular glands of the amphibian skin are due to difference in the concentrations of secretory substances as related to the processes of its formation, and that those chemical components are identical.

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The Ultrastructure of the Mucus Secreting Cells in the Amphibian Skin (양서류 피부 점액분비세포의 미세구조)

  • Kim, Han-Hwa;Noh, Yong-Tai;Chung, Young-Wha;Chi, Young-Duk
    • The Korean Journal of Zoology
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    • v.21 no.1
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    • pp.29-39
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    • 1978
  • The authors observed the ultrastructure of the mucous glandular epithelial cells in the amphibian skin by mean of electron microscope. The specimens from the experimental animals were fixed in 2.5% glutaraldehyde-oaraformaldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 1% osmium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with Sorvall MT-2 ultramicrotome. The ultrasections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows: 1. The cutaneous mucous glands in amphibia consisted of the glandular epithelial and the myoepithelial cells. 2. Several different cells in ultrastructure were observed in the mucous glandular epithelium of the adult amphibian skin. a. The dark and the light cells were observed in Hynobius leechi. b. The mitochondria-rich and the round secretory granule-containing cells were observed in Bombina orientalis. c. The round secretory granule-containing and the foam-like granule mass-containing cells were observed in Kaloula tornieri. d. The cutaneous mucous gland of Rana nigromaculata were divided into two types: A and B-type glands. In the A-type mucous gland, the mitochondria-rich and the round secretory granule-containing cells and in the B-type mucous gland, the mitochondria-rich, the secretory granule-containing and the ER-rich cells were observed. 3. Based upon the above findings, the authors infer that the mucous granular epithelium of the amphibian skin consists of the mitochondria-rich undifferentiated, the secetory granule-containing and mature, and the ER-rich evacuated cells.

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