• Title/Summary/Keyword: Histochemical method

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Histochemical Study on the Hemocytes during Metamorphosis (배추흰나비의 變態에 따른 血球細胞의 組織化學的 硏究)

  • Kim, Chang-Whan;Kim, Woo-Kap;Kim, Yong-Kuk
    • The Korean Journal of Zoology
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    • v.11 no.4
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    • pp.103-117
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    • 1968
  • Insect hemocytes during metamorphosis were studies by histchemical and autoradiographic methods is Pieris rapae (Lepidoptera). The hemocytes were classified into six types, prohemocytes, plasmatocytes, podocytes, granular hemocytes, spherule cells and oenocytoids on the basis of the transitions in shapes and inclnsions of cytoplasms. Proteins, nucleic acids, polysaccharides and lipids in the hemocytes were detected histochemically from larval to pupal stages to learn the rise and fall of them during growth and metamorphosis. Most of the granules consisted of glycogen, neutral mucopolysaccharides and mucoprotein in addition to some granules of neutral fats and phospholipids were found in the granular hemocytes and spherule cells. Mitotic figures and DNA synthetic activities were observed in every type of hemocytes from 2nd to 5th instars, suggesting the all types of hemocytes originated from the prohemocytes. The cytoplasmic filaments of plasmatocytes and pdocytes extended very long in prepupa and pupa and the vermiform cells were the transformed plasmatocytes due to their further differentiation.

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Dose Dependent Effects of Epimedii Herba on the Reproductive Function in the Mice (농도별(濃度別) 음양곽(淫羊藿) 투약(投藥)이 수컷 생쥐의 생식능력(生殖能力)에 미치는 영향(影響))

  • Lee, Chang Hoon;Cho, Jung-Hoon;Jang, Jun-Bock;Lee, Kyung-Sub
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.1
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    • pp.142-155
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    • 2005
  • Objective: This study was undertaken to evaluate the dose dependent effects of Epimedii Herba extract solution on the spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testis, and the activities of spermhyaluronidase and antioxidants. Materials and Method: We choose the 2-month-old mice, and administered the extract powder of Epimedii Herba in the different concentration once in a day for 60 days. The control group was administerde to normal water in the isolated testis tissue. Also we observed changes of isolated testis at the before and after administration of Epimedii Herba extracts in the mice. And we compared to the testicular tissue especially seminiferous tubules between control and treated group by histochemical methods. Results: The significant dose dependent differences were observed in the concentration of total sperm, the motility and normality of spermatozoa of the Epimedii Herba extract administered groups compared than that of control group, respectively. In the histological analysis of the testicular lobes were observed in the Epimedii Herbaextract administered groups than control group, respectively. Also, the activity hyluronidase was significantly increased in the Epimedii Herba extract administered groups than that of the control group. In case of antioxidant activity analysis, the activity of peroxidase and catalase were significantly increased in the Epimedii Herba extract administered groups than that of control group, respectively. Conclusion: This study shows that Epimedii Herba can effect the count and motility of sperm, the important ractor in male fertility and also promote the activity of antioxidants, catalase and peroxidase, which is the important factor in spermatogenesis.

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Totipotential, Morphological, Biochemical Comparisons between Nonembryogenic Callus and Embryogenic Callus in Water Dropwort(Oenanthe stolonifera DC) (미나리에서 비배발생캘러스와 배발생캘러스간의 분화능력 및 해부학적, 생화학적 특성비교)

  • 빈철구;김병동
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.3
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    • pp.167-173
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    • 1997
  • The embryogenic callus (EC), from which somatic embryos could be induced, was compared with nonembryogenic callus(NE) to study the origin and features of totipotent cell in water dropwort (Oenanthe stolonifera DC). To induce and maintain of EC and the NE, meristematic stem and immature floret were inoculated in MS media supplemented with 1 mg/L 2,4-D, and with 2.5 mg/L NAA and 5mg/L BA, respectively, The EC was not induced from the NE even after subculturing in MS medium supplemented with 1 mg/L 2,4-D. Plantlets were not regenerated from the NE in hormone-free medium. In histochemical comparison of the EC with the NE by light microscopy, the EC had smaller cells in size, dense cytoplasm, and more starch granules of cells compared to the NE cells. The cell from the EC, as observed by transmission electron microscopy, had smaller vaculoes, well developed ribosomes, mitochondria, and endoplasmic reticulum, whereas the cells from the NE had larger vacuoles and underdeveloped organelles. In protein pattern from NE, EC and Somatic embryo (SE), as analyzed by SDS polyacrylamide gel electrophoresis, different proteins specific for tissue were observed: 17 and 28 KD for NE, 50, 52, 57, 66, 68 KD for EC and 20 KD for SE. DNA polymorphism was also observed between EC and NE as analyzed by RAPD (randomly amplified polymorphic DNA) method. The origin of totipotent stem cell and the relationship between irreversible genomic change arose in differentiation and the loss of totipotency in plant were discussed.

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Effects of Electroacupuncture by Different Frequencies and Treatment-times on NADPH-diaphorase and Neuropeptide Y in Sprague-Dawley Rat Cerebral Cortex (전침자극이 흰쥐 대뇌피질의 NADPH-diaphorase와 NPY에 미치는 영향)

  • Cho, Seong-Gyu;Kim, Chang-Whan;Kim, Yong-Suk
    • Journal of Acupuncture Research
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    • v.19 no.3
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    • pp.95-106
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    • 2002
  • The present study was designed to investigate the effects of various electroacupuncture(EA) stimulation on NADPH-diaphorase and Neuropeptide Y(NPY) in the cerebral cortex of Sprague-Dawley Rats. We evaluated the changes of NADPH-d-positive neurons using a histochemical method and the changes of NPY-positive neurons using a double labeling immunohistochemical method. The staining intensities of NADPH-d-positive neuron and NPY-positive neurons were assessed in a quantitative fashion using a microdensitometrical method based on optical density by means of an image analyzer. As to frequency, the optical density of NADPH-d-positive neurons of 2Hz-1 time EA group was significantly higher than that of 100Hz-1 time EA group in M1, Cg, and that of 100Hz-3 times EA group was significantly higher than that of 2Hz-3 times EA group in Vi, Au, Cg and Ins. As to treatment-time, the optical density of NADPH-d-positive neurons of 2Hz-1 time EA group was higher than 2Hz-3 times EA group in Vi, Au, Cg and Ins. And in Vi 100Hz-1 time EA group and in M1 100Hz-3 times EA group was more efficient. As to frequency, the optical density of NPY-positive neurons of 2Hz-1 time was significantly higher in Vi, and that of 100Hz-1 EA group was higher in M1. And that of 2Hz-3 times EA group was higher than 100Hz-3 times EA group in Cg. As to treatment-time, the optical density of NPY-positive neurons of 2Hz-1 time EA group was significantly higer than 2Hz-3 times EA in S1, Vi, Au. And that of 100Hz-1 time EA group was significantly higher than that of 100Hz-3 time EA in Cg. The present results demonstrated that EA changes the activity in NO and NPY system in the cerebral cortex of SHR and EA stimulation, like frequency and treatment-time, is of importance for this effect.

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The effect of a static magnetic field of Nd-Fe-B magnet on alkaline phosphatase activity of MC3T3-E1 cells (Nd-Fe-B 자석의 정자기장이 MC3T3-E1 세포의 alkaline phosphatase 활성도에 미치는 영향)

  • Kim, Sook-Hee;Kwon, Oh-Won;Ryoo, Hyun-Mo
    • The korean journal of orthodontics
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    • v.30 no.2 s.79
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    • pp.205-214
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    • 2000
  • Recently, the magnetic force has been considered as a method for a more efficient tooth movement. The purpose of this study was to evaluate the effects of different static magnetic fields of Nd-Fe-B magnet on MC3T3-E1 cells by measuring the alkaline phosphatase activity and observing the amount of stained alkaline phosphatase. For measuring of alkaline phosphatase activity, MC3T3-E1 cells were seeded in first and third row of 12 well culture plates. And Nd-Fe-B magnets were positioned under the first column of first and third row to apply different static magnetic fields(first column:100mT ; second column:4.6mT ; third column:0.5mT ; forth column:0.0mT) to the cells for 7, 13, 19, and 25 days. For staining of alkaline phosphatase, MC3T3-E1 cells were seeded in 100mm culture plates. And Nd-Fe-B magnets were positioned under the corner of plates to apply different static magnetic fields(magnet side:100mT : the opposite side:0.5mT) to the cells for 7, 13, 19, and 25 days. The results were as follows : 1. ALP activity was increased until day 19 in biochemical determination as well as in histochemical staining, 2. The application of higher magnetic field(100mT) suppressed ALP activity at day 13, 19, 25. On the contrary, the application of the lower magnetic field(4.6mT, 0.5mT) significantly enhanced the ALP activity. 3. Consistent with enzyme assay, histochemical staining of ALP also demonstrated that higher magnetic field(100mT) suppressed ALP activity, lower one(0.5mT) enhanced.

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Detection of Mycobacterium Tuberculosis by In Situ Hybridization (조직내교잡법을 이용한 결핵균의 검출)

  • Park, Chang-Soo;Kim, Young-Chul;Lee, Jee-Shin;Jung, Jong-Jae;Kim, Doo-Hong;Kim, Jin
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.5
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    • pp.699-708
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    • 2000
  • Background : A presumptive histopathologic diagnosis of tuberculosis is commonly based on the finding of acid-fast bacilli upon microscopic examination of a diagnostic specimens. Although this traditional histochemical staining method is satisfactory, it is time-consuming and not species-specific. For more specific assessment, in situ hybridization assay with oligonucleotide probes is introduced. Methods : The human surgical specimens were obtained from tuberculosis patients, and experimental specimens were made by injecting cultured M. tuberculosis organisms into fresh rat liver. Oligonucleotide probes complementary to ribosomal RNA portion were synthesized and labeled with multiple biotin molecules. For a rapid detection, all procedures were carried out using manual capillary action technology on the Microprobe staining system. Results : The in situ hybridization assay produced a positive reaction in experimental specimens (80-90% sensitivity) after pepsin-HCl pre-treatment for a good permeabilization of probes, but reliable result was not obtained from human surgical specimens. Conclusion : It is, therefore, suggested that biotin-labeled oligonucleotide probes have considerable potential for identification and in situ detection of M. tuberculosis but, there are some barriers to overcome for the diagnostic use of this method.

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Fixation and Histochemistry of Biological Tissues Using the Microwave Fixator Equipped with Infrared-Temperature Sensor (적외선 온도감응기를 장착한 마이크로파 고정기에 의한 생체조직 고정효과와 조직화학적 특성)

  • 신길상;민소연;김완종;손태호
    • The Korean Journal of Zoology
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    • v.38 no.3
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    • pp.417-425
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    • 1995
  • The present study was carried out to investigate the effect of microwave fixation in comparison with that of chemical fixation in preparing the microscopic samples. The microwave fixator was equipped with infrared-temperature sensor, and that was designed to compensate air temperature in the microwave fixator. In the microwave fixation, rat tongue was well preserved in terms of muscular fasciculus and pancreas stained by Feulgen reagents showed clear reaction products in the nucleus. Reaction products by PAS method in duodenal villi appeared specifically at the goblet cells. In electron microscopy, pancreatic cellular components such as secretory granules and collagen bundles were well preserved in both fixations. In aspect of histochemical reaction and electron microscopy, high quality was due to the protein content of microwave fixed specimen. The microwave fixation method saved total duration engaging microscopic preparation.

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Central Neural Pathways Projecting to the Acupoints B62 and K6 Using the Neural Tracer (신경추적자를 이용한 신맥과 조해에서 투사되는 신경원의 표지부위에 대한 연구)

  • Kim, Su-Hyun;Lee, Chang-Hyun;Yuk, Sang-Won;Lee, Kwang-Gyu;Lee, Tae-Young;Lee, Sang-Ryoung
    • Journal of Acupuncture Research
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    • v.18 no.2
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    • pp.51-66
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    • 2001
  • The purpose of this morphological studies was to investigate the central neural pathway projecting to the acupoints $B_{62}$ and $K_6$ using the neuroanatomical method following injection of transsynaptic neurotropic virus, pseudorabies virus(PRV-Ba and PRV-Ga) into the $B_{62}$ and $K_6$. After survival times of 96 hours following injection into the twenty rats with PRV-Ba(Bartha strain) and PRV-Ga(Bartha strain, ${\beta}$-galacidodase insertion). They were perfused, and their spinal cord and brain were frozen sectioned($30{\mu}m$). These sections were stained by X-gal histochemical and PRV immunohistochemical staining method, and observed with light microscope. The results were as follows : 1. In spinal cord, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in thoracic, lumbar and sacral spinal segments. In thoracic spinal segments, Densely labeled areas were founded in lamina IV, V, VII(intermediolateral nucleus) and X areas. In lumbar segemnts, labeled areas were founded in lamina II, IV, V and X areas. In sacral spinal segments, labeled areas were founded in lamina IV, V and VI areas. 2. In brain, overlaped PRV-Ba and PRV-Ga labeled neurons projecting to the $B_{62}$ and $K_6$ were founded in the $A_1$ noradrenalin cells/$C_1$ adrenalin cells/caudoventrolateral reticular nucleus, rostroventrolateral reticular nuclens, nucleus tractus solitarius, area postrema, raphe obscurus nucleus, raphe paltidus nucleus, raphe magnus nucleus, lateral paragigantoceltular nucleus, lateral rcticular nucleus, gigantocellular nucleus, locus coeruleus, subcoeruleus nucleus, motor trigeminal nucleus, Kolliker-Fuse nucleus, $A_5$ cell group, central gray matter, oculomotor nerve, paraventricular hypothalamic nucleus, median eminence, amygdaloid nucleus, frontal cortex, forelimb area, hindlimb area, 1, 2 areas of parietal cortex and granular and agranular cortex. This results were suggest that overlaped PRV-Ba and PRV-Ga labeled areas projecting to the $B_{62}$ and $K_6$ may be related to the emotional relay pathway in the central autonomic center.

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Study of the Function of β-D-fructofuranosidase by Plant Histological Method (식물 조직학적 방법에 의한 β-D-fructofuranosidase의 기능 연구)

  • Donggiun Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.4
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    • pp.587-592
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    • 2023
  • Plant growth is regulated by a variety of factors, including organic matter availability. Organic nutrients are carbohydrate molecules from photosynthetic products produced by tissues associated with carbon and energy fixation called "sources". These compounds flow through plant vascular bundles into non-photosynthetic or growing tissues called "sinks". Among these possible compounds, the disaccharide fructosyl glucose, sucrose, is the most representative. During the transport of sucrose, the pathway from the source to the sinks can include hydrolysis of sucrose into glucose and fructose derivatives or direct transfer of sucrose. Among the enzymes involved in this, β-D-fructofuranosidase is the most important. Soluble neutral β-D-fructofuranosidase, one of several isoenzymes, is located in intracellular protoplasts and helps plant cells metabolize sucrose to produce energy. In order to track the activity of this enzyme during the course of plant growth, histological methods were used for the most effective immunolocalization. As a result, the activity was higher in the phloem and epidermis than in the mesophyll tissue in the leaf. In the growing stem, activity was high in the phloem, epidermis, and cortex. The activity of the root, which is a sink tissue, was high in all parts, but especially the highest in the root tip part. It is thought that this is because it helps unloading of sucrose in sink tissues that require sucrose degradation and plays a role in hydrolysising sucrose.

Effect of Pyrroloquinoline Quinone on Osteoclast Generation and Activity (Pyrroloquinoline quinone이 파골세포의 생성 및 활성에 미치는 영향)

  • Ko, Seon-Yle;Han, Dong-Ho;Kim, Jung-Keun
    • Journal of Oral Medicine and Pain
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    • v.30 no.3
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    • pp.329-336
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    • 2005
  • We examined the effect of PQQ, as a scavenger of superoxide, on osteoclast-like cell formation and on mature osteoclast function. To determine whether PQQ scavenges the superoxide, nitroblue tetrazolium (NBT) staining, which is a method to detect superoxide, was performed on HD-11 cells which are a chick myelomonocytic cell line having tartrate-resistant acid phosphatase (TRAP) activity in response to 1,25-dihydroxyvitamin $D_3\;[1,25(OH)_2D_3]$. Histochemical study of TRAP was also performed on HD-11 cells. PQQ inhibited the TRAP-positive multinucleated cell formation of chicken bone marrow cells was also examined. The addition of 20 ${\mu}M$ PQQ inhibited the formation of TRAP-positive multinucleated cell. When chicken osteoclasts were cultured on dentin slices, treatment of 20 ${\mu}M$ PQQ resulted in a significant decrease in dentin resorption by osteoclasts in terms of total resorption area and number of resorption pits. The present data suggest that PQQ, possibly as a scavenger of superoxide ion, inhibits the osteoclastic differentiation and bone resorption.