• Title/Summary/Keyword: Histochemistry

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Modulation in NMDA and $GABA_A$ Receptor Expression after Cerebroventricular Infusion of Ginsenosides

  • Oh Seikwan;Kim Hack-Seang
    • Proceedings of the Ginseng society Conference
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    • 2002.10a
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    • pp.96-112
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    • 2002
  • In the present study, we have investigated the effects of centrally administered ginsenoside Rc or Rgl on the modulation of NMDA receptor and $GABA_A$ receptor binding in rat brain. The NMDA receptor binding was analyzed by quantitative autoradiography using $[^3H]MK-801$ binding, and $GABA_A$ receptor bindings were analyzed by using $[^3H]muscimol\;and\;[^3H]flunitrazepam$ in rat brain slices. Rats were infused with ginsenoside Rc or Rg1 ($10\;{\mu}g/10{\mu}l/hr$, i.c.v.) for 7 days, through pre-implanted cannula by osmotic minipumps (Alzet, model 2ML), The levels of $[^3H]MK-801$ binding were highly decreased in part of cortex and cingulated by ginsenoside Rc and Rgl. The levels of $[^3H]muscimol$ binding were strongly elevated in almost all regions of frontal cortex by the treatment of ginseoside Rc but decreased by ginsenoside Rg 1. However, the $[^3H]flunitrazepam$ binding was not modulated by ginsenoside Rc or ginsenoside Rgl infusion. These results suggest that prolonged infusion of ginsenoside could differentially modulate $[^3H]MK-801\;and\;[^3H]muscimol$ binding in a region-specific manner. Also, we investigated the influence of centrally administered ginsenoside on the regulation of mRNA levels of the family of NMDA receptor subtypes (NR1, NR2A, NR2B, NR2C) by in situ hybridization histochemistry in the rat brain. The level of NR1 mRNA is significantly increased in temporal cortex, caudate putamen, hippocampus, and granule layer of cerebellum in Rgl-infused rats as compared to control group. The level of NR2A mRNA is elevated in the frontal cortex. In contrast, it was decreased in CAI area of hippocampus in Rgl-infused rats. However, there was no significant change of NR1 and NR2A mRNA levels in Rc-infused rats. The level of NR2B mRNA is elevated in cortex, caudate putamen, and thalamus in both Rc- and Rg-infused rats. In contrast, NR2B level is decreased in CA3 in Rgl-infused rats. The level of NR2C mRNA is increased in the granule layer of cerebellum in only Rg1 but not Rc infused rats. These results show that structure difference of ginsenoside may diversely affect the modulation of expression of NMDA receptor subunit mRNA after infusion into cerebroventricle in rats.

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Effects of Dangguijakyaksan on the Development of Experimentally-induced Endometriosis and Related Cytokines in Rats (당귀작약산(當歸芍藥散)이 실험적으로 유발된 흰쥐의 자궁내막증의 발달 및 관련 Cytokine에 미치는 영향)

  • Joo, Byoung-Ju;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.4
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    • pp.104-127
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    • 2008
  • Purpose: This study was performed to investigate the effects of Dangguijakyaksan on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group(n=8). (2) surgically induced endometriosis and untreated control group(n=8). (3) surgically induced endometriosis and Dangguijakyaksan treated group. Dangguijakyaksan was orally administrated for 15 days after operation. Then we measured the body weight. the volume of endometriotic implants. the weight of uterus and ovary. and analysed the concentration of cytokines (MCP-1, TNF-$\alpha$, IL-1$\beta$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF. and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: - The volume($mm^3$) of endometriotic implants in Dangguijakyaksan treated group was significantly decreased compared with control group. - The concentration(pg/ml) of MCP-1, TNF-$\alpha$ and IL-l$\alpha$ in peritoneal fluids in Dangguijakyaksan treated group was significantly decreased compared with control group. - Histopathologically, proliferation of endometriotic epithelia. infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The percentage of positive epithelial layers for COX-2 in Dangguijakyaksan treated group was significantly decreased compared with control group. - The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Dangguijakyaksan treated group were weakly observed than those of control group. - The number of mast cells in transplanted uterine tissue in Dangguijakyaksan treated group was significantly decreased compared with control group. Conclusion: On the basis of these results. we concluded that Dangguijakyaksan has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-$\alpha$ and IL-1$\beta$, and decreased expression of COX-2 and VEGF, and inhibition of mast cell infiltration by administration of Dangguijakyaksan.

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Effects of Hominis Placenta Pharmacopuncture Therapy on the Experimentally-induced Endometriosis in the Rats (자하거(紫河車) 약침(藥鍼)이 실험적으로 유발된 흰쥐의 자궁내막증에 미치는 영향)

  • Yoo, Yung-Ki;Kim, Hyung-Jun;Sin, Mi-Ran;Lee, Dong-Nyung
    • The Journal of Korean Obstetrics and Gynecology
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    • v.27 no.1
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    • pp.101-119
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    • 2014
  • Objectives: This study was performed to investigate the effects of Hominis Placenta pharmacopuncture (HPP) therapy on the experimentally-induced endometriosis in the rats. Materials and Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and divided them into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group (n=8), (3) surgically induced endometriosis and HPP treated group. Sham-operated group and control group were inject with normal saline once a every other day for 30days, while treated group was injected with HPP extract once a every other day for same duration. Injected point of HPP and normal saline were subcutaneous tissue at Gwanwon (CV4) acupoint. Then we measured the body weight, the volume of endometriotic implants, the weigh of uterus and ovaries, and investigated the concentration of cytokines (MCP-1, TNF-${\alpha}$) in peritoneal fluids. Histopathology, immunohistochemisty for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were performed. Results: The volume ($mm^2$) of endometriotic implants in HPP treated group ($55.4{\pm}41.6$) was significantly decreased (p<0.01) compared with control group ($140{\pm}66.1$). And the concentration (pg/ml) of MCP-1 in peritoneal fluids in HPP treated group ($1117.6{\pm}60.5$) was significantly decreased (p<0.01) compared with control group ($1446.2{\pm}280.3$). The concentration (pg/ml) of TNF-${\alpha}$ in peritoneal fluids in HPP treated group ($80.6{\pm}31.4$) was decreased (p<0.01) compared with control group ($145.3{\pm}86.9$). Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cell and angiogenesis in transplanted uterine tissue of HPP treated group were weakly observed than those of control group. The COX-2 expression in endometrial, epithelial and stromal cells in transplanted uterine tissue of HPP treated group was decreased compared with control group. The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cell in transplanted uterine tissue of HPP treated group were weakly observed than those of control goup. Conclusions: HPP is effect on Endometriosis of rats by Experimentally-induced.

A Comparative Study on the Effects of Pinellia ternata, Zingiber officinale and Sobanhatang on Reflux Esophagitis (역류성 식도염에 대한 반하(半夏), 생강(生薑), 소반하탕(小半夏湯)의 효과 비교)

  • Lee, Seul-hee;Baik, Tai-Hyeun
    • The Journal of Korean Medicine
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    • v.40 no.2
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    • pp.17-34
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    • 2019
  • Objectives: This study was carried out to observe and compare the effects of Pinellia ternata, Zingiber officinale and Sobanhatang on the reflux esophagitis induced by gastric fundus and pylorus ligation in mice with esomeprazole. Methods: Antioxidant effects were measured by DPPH radical scavenging activity at four different concentration of 0.125, 0.25, 0.5 and $1.0mg/102{\mu}{\ell}$. Zingiber officinale water extract(ZE), Pinellia ternata water extract(PE) and Sobanhatang water extract(SBE) and esomeprazole were treated orally for 14 days before gatric fundus and pylorus ligation. In the histochemistry, changes in suface mucous cells, muscle tissue and connective tissue in gastro esophageal junction(GEJ) and mast cell on the esophageal mucosa were observed. The change of Hemo oxygenase(HO)-1, ghrelin, gastrin and substance P in gastric body tissue were measured by immunohistochemistry. Results: DPPH radical scavenging activity exhibited concentration dependently increases in ZE, PE, SBE. ZE was significantly higher at all concentrations than PE. The gastric surface mucous cells were more in the treated group than in the reflux esophagitis elicited group(GE) in the order of PE, SBE, ZE and esomeprazole treateded group(PT, SBT, ZT, ET). Lower esophageal sphincter muscle damage and intercellular space in the GEJ were less in the treated group than GE. In the esophageal mucosa, the mast cell distribution and the migration of inflammatory cells were lower in the treateded troup than GE in order to ZT, SBT, PT and ET. The antioxidative enzyme, HO-1 was more in the order of ZT, SBT, control group, PT, ET than in GE. ZT was significantly higher than the other groups and SBT was significantly higher than ET. Ghrelin was found to be higher in ZT, ET, SBT and PT than in GE, and ZT was significantly higher than all other groups except ET. Gastrin showed the highest positivity in GE, and was lower in the order of ET, ZT, SBT, PT, and control group. Substance P was the highest in GE, and was lower in the order of ET, ZT, SBT, PT and control group, and PT were significantly lower than ET. Conclusion: ZT, PT and SBT showed superior antioxidative, anti-inflammatory and mucosal protective effects on mouse reflux esophagitis as compared with ET. In particular, ZE was more effective in antioxidant and gastric motility enhancement, while PE was more effective in mucosal protection and anti-inflammatory effects. Sobanhatang is expected to be effective treatment because it has advantages of both drugs and reduces toxicity.

Effects of Bojungikgi-Tang on the Development of Experimentally-induced Endometriosis in the Rats (보중익기탕(補中益氣湯)이 실험적으로 유발된 흰쥐의 자궁내막증(子宮內膜症) 발달에 미치는 영향)

  • Lee, Jeong-Gu;Lee, Dong-Nyung;Seo, Il-Bock;Kim, Hyung-Jun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.3
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    • pp.1-24
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    • 2009
  • Purpose: This study was performed to investigate the effects of Bojungikgi-Tang on the development of experimentally-induced endometriosis in rats. Methods: Endometriosis was induced in rats by autotransplanting uterine tissue to the peritoneum and we divided the rats into three groups: (1) sham-operated group (n=8), (2) surgically induced endometriosis and untreated control group(n=8), (3) surgically induced endometriosis and Bojungikgi-Tang treated group. Bojungikgi-Tang was orally administrated for 15 days after operation. Then we measured the body weight, the volume of endometriotic implants, the weight of uterus and ovary, and analysed the concentration of cytokines(MCP-1, TNF-${\alpha}$, IL-1${\beta}$, IL-6) in peritoneal fluids. Histopathology, immunohistochemistry for COX-2 and VEGF, and histochemistry for mast cell in transplanted uterine tissue were also performed. Results: 1. The volume($mm^3$) of endometriotic implants in Bojungikgi-Tang treated group(129${\pm}$39) was significantly decreased(p<0.05) compared with control group(222${\pm}$109). 2. The concentration(pg/$m{\ell}$) of MCP-1 in peritoneal fluids in Bojungikgi-Tang treated group(1012${\pm}$248) was significantly decreased(p<0.05) compared with control group(1413${\pm}$346). 3. The concentration(pg/$m{\ell}$ of TNF-${\alpha}$ in peritoneal fluids in Bojungikgi-Tang treated group(887${\pm}$196) was significantly decreased(p<0.05) compared with control group(1126${\pm}$140). 4. Histopathologically, proliferation of endometriotic epithelia, infiltration of inflammatory cells and angiogenesis in transplanted uterine tissue of Bojungikgi-Tang treated group were weakly observed than those of control group. 5. The percentage of positive epithelial layers for COX-2 in Bojungikgi-Tang treated group(56${\pm}$17) was significantly decreased(p<0.05) compared with control group(75${\pm}$16). 6. The VEGF expression of endometriotic epithelia, neovascular endothelia and stromal cells in transplanted uterine tissue of Bojungikgi-Tang treated group were weakly observed than those of control group. Conclusion: On the basis of these results, we concluded that Bojungikgi-Tang has inhibiting effects on the development of transplanted uterine tissue. And these effects may be related with decreased production of MCP-1, TNF-${\alpha}$, and decreased expression of COX-2 and VEGF infiltration by administration of Bojungikgi-Tang.

Histology and Histochemistry of the Male and Female Reproductive System of the Sesarmid Crab Muradium Tetragonum

  • Pothiappan Kumarasamy;Viswambaram Ganapiriya;Kannayiram Muthukumaravel;Manickam Sasipriya;Bharathi Santhanabharathi;Mohamed Saiyad Musthafa;Marckasagayam Priyadharshini
    • Journal of Integrative Natural Science
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    • v.16 no.1
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    • pp.13-31
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    • 2023
  • The sesarmid crab Muradium tetragonum, inhabiting the mangrove, are considered as a key consumer of litter and thereby play an important role in the detritus food chain and energy flow in the mangrove ecosystem. The present investigation was carried out with objectives to enlighten the reproductive system of Muradium tetragonum through histological and histochemical studies. Histological organization of the testis of M. tetragonum revealed that each testis has a lobular structure consisting of several testicular lobules arranged around the collecting duct. Histology of the deferens of M. tetragonum revealed it to be composed of three-layer of tissues along the entire length:the outer connective tissue, the middle muscular and the inner epithelial layer. Based on the histological architecture these three regions are recognized as proximal vas deferens (PVD), middle vas deferens (MVD) and distal vas deferens (DVD). Histological characteristics of the ovary of M. tetragonum during different phases of ovarian development were studied. Based on the colour changes of the ovary and diameter of the oocytes five stages of ovarian development can be pronounced. Histochemical analysis of the male reproductive tissues of M. tetragonum signifies the secretion of a different biomolecule by specifying their origin in the reproductive tissue and their possible transformation into spermatophores. In the female reproductive tissues, histochemical evaluation envisaged the secretory products during different stages of ovarian development The secretory substances of the spermatheca expound on the significance of its secretion in dehiscing the spermatophore wall and in nourishing as well as protecting the spermatozoa.

Developmental Changes in Histology and Histochemistry of Epididymal Ducts in Korean Native Goats II. Histochemical changes in epididymal ducts (한국재래산양 정소상체관의 발육에 따른 조직 및 조직화학적 변화 II. 정소상체관의 조직화학적 변화)

  • 김성호;김창근;정영채;이재홍;이방환
    • Korean Journal of Animal Reproduction
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    • v.11 no.3
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    • pp.196-205
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    • 1987
  • This study was conducted in order to obtain the informatin of the histochemical changes in each of 6 segments of the epididymal ducts in 32 Korean native male goats. The male goats were examined, dividing into 7 groups, at 4 잔 intervlas from 8 to 32 wks of age. The reuslts obtained were as follows: 1. PAS reaction showed positive on the basal and upper part beyond the nucleus of the peithelium of effernt ductules throughout all the classes of age. It was also positive on the free border and basal and upper part beyond the nucleus of the caput, on the free border andbasal parts of the corpus, and on the basal part of the cauda of the epididymal epithelium. 2. Acid phosphatase reaction was negative on the every part of epididymal epithelium at the age of 8 weeks, however, with the aging it became strangly positive on the areas between the free border and the nucleus, and moderately positive on the basal part of epithelium of the caput and corpus. In the free border adn basal part of the cauda, it was slightly positive. alkaline phosphatase reactin was negative on the every part of epididymal epithelium until 12 weeks of age. From 16 weeks, free border of epididymal epithelium becaqme slightly positive, and from 20 weeks, the reaction became stronger on the basal part but weekend on the free border with the aging. 3. In the sudan black B staining, many blue black granules between the free border and the nucleus, some granules on the basal part, and a few granules on the cytoplasm around the nucleus of the epididymal epithelium were observed from 8 weeks of age as early, and the granules were increased in number with the aging. 4. In Azan staining, reddish violet granules below the nucleus and blue granules on the upper part beyond the nucleus in some cells of epithelia of efferent ductules were noted at 12th and 16th week, and after 24th week, the granules were decreased with the aging. Golgi apparatus were clearly observable on the upper part beyond the nucleus of all parts of epididymal epithelium from 8th week, and also number of intracytoplasmic vacuoles(smaller ones on the upper part and larger ones on the basal part beyond the nucleus) and fine granules were increased with the aging. 5. In the toluidin blue staining, reddish purple granules on the basal part of the epithelium in all the parts of epididymal ducts, particularly brilliant in the cauda, were observed from 8th week as early. 6. In the Cowdry staining, numerous mitochondria, according to aging, were observed between the free border of epithelium and the upper part beyond the nucleus particularly in the catus and corpus of the epididymal ducts.

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The Effect of Low Dose Radiation on the Neuronal Cell Proliferation in Diabetic Rats (당뇨병 유발 쥐에서 신경 세포 증식에 대한 저선량 방사선의 효과)

  • Kim Doo Soon;Kang Jin Oh;Hong Seong Eon;Kim Sang Ki;Lee Taeck-Hyun;Kim Chang-Ju
    • Radiation Oncology Journal
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    • v.23 no.3
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    • pp.157-160
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    • 2005
  • Purpose: To investigate the effect of low dose radiation on neuronal cell proliferation In diabetic rats. Materials and Methods: A group of rats (first group) were divided into three subgroups (nondiabetic control, nondiabetic 0.1 Gy and nondiabetic 10 Gy groups) to determine the effect of radiation on normal hippocampal neuronal ceil proliferation. A further group of rats (second group) were divided into six subgroups (nondiabetic control, diabetic control, diabetic 0.01 Gy, diabetic 0.1 Gy, diabetic 1 Gy and diabetic 10 Gy groups) to determine the effect of radiation on hippocampal neuronal cell proliferation under diabetic conditions. Using immuno-histochemistry for 5-bromo-2'-deoxyuridine (BrdU), the number of neuronal cells in the dentate gyrus of all the groups was counted. Results: The number of BrdU-positive cells in the dentate Gyrus of the nondiabetic control, nondiabetic 0.1 Gy and nondiabetic 10 Gy subgroups of the first group were $45.95{\pm}3.42,\;59.34{\pm}5.20\;and\;19.26{\pm}2.98/mm^2$, respectively. The number of BrdU-positive cells in the dentate gyrus of the diabetic control, diabetic 0.01 Gy, diabetic 0,1 Gy, diabetic 1 Gy and diabetic 10 Gy subgroups of the second group were $55.44{\pm}8.57,\;33.33{\pm}6.46,\;67.75{\pm}10.54,\;66.63{\pm}10.05,\;23.59{\pm}6.37\;and\;14.34{\pm}7.22/mm^2$, respectively. Conclusion: Low dose radiation enhances cell proliferation in the dentate gyrus of STZ-induced diabetic rats.

Structure and Metallothionein Expression during Rat Liver Regeneration Induced by Partial Hepatectomy (흰쥐 부분 간 절제 후 재생 중인 간 조직의 구조와 metallothionein 분포)

  • Mun, Seung-Hoon;Jeong, Jin-Joo;Lee, Yong-Moon;Shin, Kil-Sang;Kim, Wan-Jong
    • Applied Microscopy
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    • v.38 no.3
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    • pp.175-183
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    • 2008
  • Liver regeneration is a result of highly coordinated proliferation of hepatocytes and non-parenchymal liver cells. At this process, induction of metallothionein (MT), which is low molecular and cysteine rich, has been reported. The present study was carried to find the ultrastructure of hepatocytes and determine the expression of MT in regenerating rat liver after partial hepatectomy. As a result, the remnant liver after PH grew fast from 1 day until 7 days. Various changes were morphologically observed. Disintegration of cell plates and liver lobule appeared shortly after PH. And hepatocytes showed the rapid proliferation, characterized by high nuclear cytoplasmic ratio, weak intercellular junctional complexes, chromatin condensation, increase of ribosomes and mitochondria, and temporary increase of lipid droplets. Finally, remodeling of the liver lobule was completed through the rearrangement of blood vessels and cell plates by 7 days after PH. On histochemistry, immunoreactivity indicating the presence of MT appeared moderately throughout the cytoplasm of control rat hepatocyte. After PH, positive reactions for MT increased at the cytoplasm and the nucleus. These results suggest that the remnant liver cells immediately entered cell proliferation and increase of MT expression after PH. It is thought that MT protein might be associated with transfer of some factors needed to cell division from the cytoplasm to the nucleus for regeneration of the liver after PH.

Localization and characteristics of lactate and palate dehydrogenase in the sparganum and adult worm of Spirometra erinacei (만손열두조충에서 젖산탈수소효소와 말산탈수소효소의 분포와 특징)

  • Gwak, Gi-Hun;Jeon, Eun-U;Kim, Chang-Hwan
    • Parasites, Hosts and Diseases
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    • v.34 no.1
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    • pp.59-68
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    • 1996
  • This study investigated the enzyme histochemical localization and characteristics of lactate (LDH) and malate dehydrogenase (MDH) related with the oxidation-reduction metabolism in the sparganum and adult of 5. erinacei. By enzyme histochemical assay, activity of LDH was strong in the tegument and subtegumental muscle layers of the adult and sparganum. Activity of MDH was strong in the tegument of the sparganum and subtegumental muscle layers of the adult. However it was weak in the tegument of the adult. By electrophoresis, 45 kDa band was major and common in LDH of adults and spargana. The 150 kDa molecule was the major and common band in MDH of adults and r -spargana (from experimentally infected rats) . By isoelectrofocusing, isoelectric points (Pl) or 4 MDH isogyme from adult worm were 6.0.6.5, 6.7 and 7.1, respectively. Pl 6.0 was the major band. The active range of pH for MDH was about pH 6-8 and the optimum pH was pH 7 The effective temperature on the MDH was about $30^{\circ}C$$50^{\circ}C$ and the optimum temperature was about 40℃ in spargana md adult worm. In the stability against heat, when MDH was heated at 85℃ for 10 seconds, the activity was denatured perfectly. Maximum activity or MDH was 19.4 unit in the s-sparganum (from snakes), 24.5 unit in the r-sparganum (from rats) and 108.0 unit in the adult worm. The maximum activity was higher in adults than in spargana. The present result showed us that the nutrients absorbed through the tegument were transferred into inner tissues and were utilized as the source of metabolism. According to the habitat of the parasite, the isozymes of LDH and MDH are activated differently, and by this different activation the sparganum and adult can adapt themselves to parasitic circumstances.

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