• Title/Summary/Keyword: Tissue homogenates

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Characterization of the Repetitive Sequences Present in the ORF25 Genomic Region of Megalocytiviruses from Ornamental Fishes (관상어로부터 분리한 Megalocytiviruses에서 나타나는 ORF25 유전자 부위의 반복서열 특성 분석)

  • Jin, Ji-Woong;Nam, Jeong-Hee;Kim, Kwang-Il;Hong, Su-Hee;Byon, Ju-Young;Jeong, Hyun-Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.44 no.4
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    • pp.352-358
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    • 2011
  • The presence of ISKNV-like viruses in various freshwater ornamental fish species imported from Asia was confirmed by polymerase chain reaction(PCR) amplification of the ATPase(adenosine triphosphatase) gene. Interestingly, molecular analyses of the Open Reading Frame 25(ORF25) region of these isolates based on the ISKNV(Infectious spleen and kidney necrosis virus) genome revealed the presence of various repetitive sequences. ORF25 repeat sequence length had no effect on cumulative mortality of rock bream Oplegnathus fasciatus challenged with tissue homogenates of infected pearl gourami, Trichogaster leeri; silver gourami, Trichogaster microlepis; blue gourami, or Trichogaster trichopterus. All isolates induce cumulative mortalities after 12 days of infection, confirming that ORF25 polymorphism did not affect the pathogenicity of ornamental fish megalocytiviruses that cross infect rock bream, a seawater fish. Also, no statistically significant differences in spleen index or viral copy number in infected tissues was detected between isolates with varying ORF25 repeat sequence lengths. However, further studies are necessary to fully characterize the functional characteristics of these polymorphisms in megalocytivirus disease in ornamental fishes.

Antioxidant effects and its mechanism of Cheongsimyeonjatang in Sasang Constitutional Medicine (청심연자탕(淸心蓮子湯)의 항산화(抗酸化) 효과(效果)와 기전(機轉)에 관한 연구(硏究))

  • Han, Byeung-Sam;Bae, Young-Chun;Song, Seung-Yun;Park, Hye Sun;Lee, Jae-Heung;Kim, Kyung-Yo
    • Journal of Sasang Constitutional Medicine
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    • v.16 no.1
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    • pp.130-147
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    • 2004
  • To investigate the antioxidant capacity of traditional Korean herb medicines, water extracts from 42 species which has been used for the Taeum constitution of Sasang Medicine, were tested on their antioxidant activity using radical scavenging effects against ABTS. Some Of them showed strong antioxidant capacities at $50{\mu}g$ concentration. And, antioxidant capacities of 2 prescriptions extract of Sasang Medicine, Cheongsimyeonja-tang and Yeoldahanso-tang, in the different concentration ($10{\mu}g$, $50{\mu}g$, and $100{\mu}g$) were determined. At the same time, the antiperoxidation effects of these 2 prescriptions extract were determined. Lipid peroxidation in brain homogenates induced by NADPH and $ADF-Fe^{2+}$ was strongly inhibited by Cheongsimyeonja-tang in vitro. The above-mentioned 2 prescriptions are a potent antioxidant capacity and antiperoxidation activity, further investigation into the in vivo antioxidant therapeutic potential for treatment of human disorders in brain tissue.

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Pomegranate (Punica granatum) Peel Extract Efficacy as a Dietary Antioxidant against Azoxymethane-Induced Colon Cancer in Rat

  • Waly, Mostafa I.;Ali, Amanat;Guizani, Nejib;Al-Rawahi, Amani S.;Farooq, Sardar A.;Rahman, Mohammad S.
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.4051-4055
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    • 2012
  • Functional foods include antioxidant nutrients which may protect against many human chronic diseases by combating reactive oxygen species (ROS) generation. The purpose of the present study was to investigate the protective effect of pomegranate peel extract (PPE) on azoxymethane (AOM)-induced colon tumors in rats as an in vivo experimental model. Forty Sprague-Dawley rats (4 weeks old) were randomly divided into 4 groups containing 10 rats per group, and were treated with either AOM, PPE, or PPE plus AOM or injected with 0.9% physiological saline solution as a control. At 8 weeks of age, the rats in the AOM and PPE plus AOM groups were injected with 15 mg AOM/kg body weight, once a week for two weeks. After the last AOM injection, the rats were continuously fed ad-libitum their specific diets for another 6 weeks. At the end of the experiment (i.e. at the age of 4 months), all rats were killed and the colon tissues were examined microscopically for lesions suspected of being preneoplastic lesions or tumors as well as for biochemical measurement of oxidative stress indices. The results revealed a lower incidence of aberrant crypt foci in the PPE plus AOM administered group as compared to the AOM group. In addition, PPE blocked the AOM-induced impairment of biochemical indicators of oxidative stress in the examined colonic tissue homogenates. The results suggest that PPE can partially inhibit the development of colonic premalignant lesions in an AOM-induced colorectal carcinogenesis model, by abrogating oxidative stress and improving the redox status of colonic cells.

Synthesis and Properties of 5-Aminosalicyl-taurine as a Colon-specific Prodrug of 5-Aminosalicylic Acid

  • Jung, Yun-Jin;Kim, Hak-Hyun;Kong, Hye-Sik;Kim, Young-Mi
    • Archives of Pharmacal Research
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    • v.26 no.4
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    • pp.264-269
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    • 2003
  • 5-Aminosalicylic acid (5-ASA) is an active ingredient of therapeutic agents used for Crohn s disease and ulcerative colitis. Because it is absorbed rapidly and extensively in the upper intestine, delivery of the agent specifically to the colon is necessary. We selected taurine as a colon-specific promoiety and designed 5-aminosalicyltaurine (5-ASA-Tau) as a new colon-specific prodrug of 5-aminosalicylic acid (5-ASA). It was expected that introduction of taurine would restrict the absorption of the prodrug and show additive effect to the anti-inflammatory action of 5-ASA after hydrolysis. 5-ASA-Tau was prepared in good yield by a simple synthetic route. The apparent partition coefficient of 5-ASA-Tau in 1-octanol/pH 6.8 phosphate buffer or $CHCl_3$/pH 6.8 phosphate buffer was 0.10 or 0.18, respectively, at $37^{\circ}C$. To determine the chemical and biochemical stability in the upper intestinal environment, 5-ASA-Tau was incubated in pH 1.2 and 6.8 buffer solutions, and with the homogenates of tissue and contents of stomach or small intestine of rats at $37^{\circ}C$. 5-ASA was not detected from any of the incubation medium with no change in the concentration of 5-ASA-Tau. On incubation of 5-ASA-Tau with the cecal and colonic contents of rats, the fraction of the dose released as 5-ASA was 45% and 20%, respectively, in 8 h. Considering low partition coefficient and stability in the upper intestine, 5-ASA-Tau might be nonabsorbable and stable in the upper intestine. After oral administration, it would be delivered to the colon in intact form and release 5-ASA and taurine. These results suggested 5-ASA-Tau as a promising colon-specific prodrug of 5-ASA.

Therapeutic Effect of Whole Bear Bile and Its Components against Croton Oil-Induced Rectal Inflammation in Rats

  • Park, Jeong-Sook;Yoo, Dong-Ho;Lee, In-Jeong;Roh, Eun-Mi-Ri;Kim, Young-Soo;Han, Kun
    • Biomolecules & Therapeutics
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    • v.18 no.1
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    • pp.83-91
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    • 2010
  • Bear bile has been used as a therapeutic for cerebral and coronary thrombosis, convulsion, hepatitis, jaundice, and abscess in traditional oriental medicine. In recent decades, the effects of bile acids on cancer, cholestasis, and liver injury have been investigated in many studies. In this study, we investigated the anti-inflammatory effects of whole bear bile (WBB) and its two major components, chenodeoxycholic acid (CDCA) and ursodeoxycholic acid (UDCA), on rectal inflammation in rats. Bile acids in WBB were quantitatively analyzed by HPLC. Rectal inflammation was induced in male Sprague-Dawley rats by insertion of croton oil-saturated cotton tips. WBB, UDCA or CDCA solution was orally administered to rats one hour after induction of rectal inflammation. Rats were sacrificed 4 or 24 hours after induction of rectal inflammation. The evaluation included measurement of weight and thickness of rectum and histopathologic examination of rectal tissue. Furthermore, we examined the inhibitory effect of WBB, UDCA or CDCA against NO production in LPS-stimulated RAW 264.7 cells. The contents of UDCA and CDCA in WBB were $39.26{\mu}g/mg$ and $47.11{\mu}g/mg$, respectively. WBB treatment significantly reduced the weight and thickness of rectum compared with UDCA or CDCA treatment. The inhibition of NO production by WBB, UDCA and CDCA in LPS-stimulated RAW 264.7 cells was much higher than that by the control. And, WBB treatment suppressed the induction of NO synthase in rectum homogenates. These results suggest that the anti-inflammatory effect of WBB is related to the suppression of NO synthase induction and the inhibition of NO production by UDCA, CDCA and other bile acids of WBB.

Study on the pathogenesis of the piglets experimentally infected with Korean isolate of Aujeszky′s disease virus I. Histopathologic and electron microscopic observation (Aujeszky's disease virus 국내분리주 접종자돈의 병리발생에 관한 연구 I. 병리학적 및 전자현미경적 관찰)

  • 조우영;조성환;김재훈;박최규;황의경;조부제;정운선
    • Korean Journal of Veterinary Service
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    • v.19 no.1
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    • pp.1-29
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    • 1996
  • This study was conducted to elucidate the pathogenesis of Aujeszky's disease virus(ADV) by histopathologic examination. The first Korean ADV Isolate, which was isolated from piglets with clinical signs of Aujeszky's disease in Yangsan(YS) county, Kyungnam province, was inoculated into 32 days old piglets with a dose of $10^{5.9}$$TCID_{50}/ml$ through intranasal or intramuscular route. These piglets were sacrificed at intervals of every 24hrs for 8 days. The virulence of YS strain was determined by the observation of clinical signs, gross findings, and histopathologic changes in tissues. The virus recovery test was performed from brain, spleen, lung and tonsil in cell culture. The pathogenesis of YS strain was determined by the observation of histopathologlc lesions in CNS and neuronal tracts. The major clinical signs were fever, anorexia, dyspnea, constipation, tremor, ataxia, circling movement, hindleg paralysis and salivation. The clinical signs were more severe in piglets of the group inoculated intranasally than those of the intramuscularly inoculated gorup. Lymphocytopenia was detected on day 5 to day 6 postinoculation (PI). The ADV was recovered from the tissue homogenates of tonsil, lung, spleen and cerebrum in cell culture. The highest virus titer was detected from tonsil between day 6 and day 7 PI. Reddish sublobar consolidation foci were scattered in the apical and cardiac lobes of lung. Although yellowish necrotic foci were detected in tonsil and liver, hemorrhagic lesions were mainly observed in heart, kidney and lymph nodes. Histopathologically, degeneration and necrosis of nerve cells, nonsuppurative meningoe-ncephalitis, nodular gliosis and perivascular cuffings were observed in CNS. Multifocal fibronecrotic foci were observed in lung, liver, lymph nodes and spleen. The major pathologic changes were detected in the midbrain, pons and medulla oblongata. Eosinophilic intranuclear inclusion bodies were mainly observed in epithelia and /or macrophages of tonsil, liver, lung, spleen and submandibular lymph nodes, and neurons of brain, respectively. Observation of viral particles at various stages of replication were possible from the endothelial cells of the alveolar capillaries and tonsillar crypt epithelia by transmission electron microscope.

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Distribution of the Muscarinic Cholinergic Receptors and Characterization in the Brain of Wistar Rats and Spontaneously Hypertensive Rats (SHR Strain) by Digital Autoradiography (Digital Autoradiographic System을 이용한 선천성고혈압에서의 Muscarinic Cholinergic Receptor 분포 및 특성)

  • Sohn, In;Lee, Myung-Chul;Koh, Chang-Soon
    • The Korean Journal of Nuclear Medicine
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    • v.27 no.1
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    • pp.28-34
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    • 1993
  • Using in vitro autoradiography with a digital autoradiography system and radioreceptor assay, the distribution and the binding characteristics of the muscarinic cholinergic receptors (mAChR) were studied in regions of rat brain. Radioreceptor assay revealed that mAChR could be measured with saturation binding assay in the brain and heart homogenates: No difference in Kd or Bmax of the brain or heart was found between the normal Wistar rats and SHR rats. Specific binding of $^3H$ quinuclidinyl benzilate (QNB) increased and saturation was reached by 2 hours after incubation with slide-mounted brain tissue. The distribution of mAChR was heterogeneous along the fields of brain. Affinity (Kd) of mAChR was not different significantly among cortex, hippocampus and caudate-putamen. No difference was found between normal rats and SHR strain. More receptors (Bmax) were found in the cortex and hippocampus than in the caudate-putamen in normal rats. More receptors were found in the cortex and caudate-putamen in SHR rats than in normal rats. Radioreceptor assay and digital autoradiographic analysis of affinity and number of mAChR gave the same results. With the above findings, we concluded that we could use digital autoradiographic system with $^3H$-QNB in the characterization of mAChR of rats and that the cortex and caudate-putamen of SHR strain rats have more receptors than those of normal rats.

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Application of the rpoS Gene for Species-Specific Detection of Vibrio vulnificus by Real-Time PCR

  • Kim, Dong-Gyun;Ahn, Sun-Hee;Kim, Lyoung-Hwa;Park, Kee-Jai;Hong, Yong-Ki;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1841-1847
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    • 2008
  • Vibrio vulnificus is a causative agent of serious diseases in humans, resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for the V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1 g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium, using the rpoS gene in pure cultures and in infected oyster tissues.

ARACHIDONIC ACID METABOLISM IN HYPERSENSITIVE HUMAN DENTAL PULP (지각과민성(知覺過敏性) 치아(齒牙) 치수조직(齒髓組織)의 Arachidonic Acid Metabolism에 관(關)한 연구(硏究))

  • Lee, Kyung-Hee;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.15 no.1
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    • pp.153-164
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    • 1990
  • Human dental pulps obtained from normal teeth, hypersensitive teeth and teeth with inflamed pulp were studied to measure and to compare the endogenous levels of arachidonic acid metabolites in order to see the relative activities of the different pathways involved in arachidonic acid metabolism. Pulp homogenates were incubated with $^{14}C$-arachidonic acid and lipid solvent extracts were separated by thin layer chromatography (TLC) to be analyzed by autoradiography and TLC analyzer. 1. The most significant metabolite was HETEs showing $96.9{\pm}37.8$pmol/mg tissue protein/hr in normal pulp, $169.2{\pm}76.7$ in hypersensitive pulp and $385.4{\pm}113.2$ in inflamed pulp. In normal pulp $LTB_4$, 6-keto-$PGF_{1\alpha}+PGE_2$, $TXB_2$ and unidentified metabolite were formed in decreasing order. While in hypersensitive and inflamed pulp 6-keto-$PGF_{1\alpha}+PGE_2$, $LTB_4$, $TXB_2$ and unidentified metabolite were formed in decreasing order. 2. In hypersensitive pulp only HETEs were significantly increased when compared with that in normal pulp. The levels of all the converted metabolites in inflamed pulp were significantly increased compared with those in normal pulp. In inflamed pulp, the levels of $TXB_2$ and HETEs were significantly increased compared with those in hypersensitive pulp. 3. The ratio of each metabolites to the total converted metabolites showed an increased value of $TXB_2$ and 6-keto-$PGF_{1\alpha}+PGE_2$, as the degree of inflammation was increased, while that of HETEs decreased both in hypersensitive pulp and inflamed pulp more than in normal pulp. 4. The relative amounts of the total metabolites formed in lipoxygenase pathway to cyclo-oxygenase pathway were 6.8 fold in normal pulp, 4.4 fold in hypersensitive pulp and 3.8 fold in inflamed pulp.

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Conversion of Apricot Cyanogenic Glycosides to Thiocyanate by Liver and Colon Enzymes

  • Lee, Ji-Yeon;Kwon, Hoon-Jeong
    • Toxicological Research
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    • v.25 no.1
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    • pp.23-28
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    • 2009
  • Some of the edible plants like apricot kernel, flaxseed, and cassava generate hydrogen cyanide (HCN) when cyanogenic glycosides are hydrolyzed. Rhodanese (thiosulfate: cyanide sulfurtransferases of TSTs; EC: 2.8.1.1) is a sulfide-detoxifying enzymes that converts cyanides into thiocyanate and sulfite. This enzyme exists in a liver and kidneys in abundance. The present study is to evaluate the conversion of apricot cyanogenic glycosides into thiocyanate by human hepatic (HepG2) and colonal (HT-29) cells, and the induction of the enzymes in the rat. The effects of short term exposure of amygdalin to rats have also been investigated. Cytosolic, mitochondrial, and microsomal fractions from HepG2 and HT-29 cells and normal male Spraque-Dawley rats were used. When apricot kernel extract was used as substrate, the rhodanese activity in liver cells was higher than the activity in colon cells, both from established human cell line or animal tissue. The cytosolic fractions showed the highest rhodanese activity in all of the cells, exhibiting two to three times that of microsomal fractions. Moreover, the cell homogenates could metabolize apricot extract to thiocyanate suggesting cellular hydrolysis of cyanogenic glycoside to cyanide ion, followed by a sulfur transfer to thiocyanate. After the consumption of amygdalin for 14 days, growth of rats began to decrease relative to that of the control group though a significant change in thyroid has not been observed. The resulting data support the conversion to thiocyanate, which relate to the thyroid dysfunction caused by the chronic dietary intake of cyanide. Because Korean eats a lot of Brassicaceae vegetables such as Chinese cabbage and radish, the results of this study might indicate the involvement of rhodanese in prolonged exposure of cyanogenic glycosides.