Kim, Sung-Joon;Park, Sung-Min;Kang, Hee;Shim, Bum-Sang;Kim, Sung-Hoon;Choi, Seung-Hoon;Ahn, Kyoo-Seok
Journal of Physiology & Pathology in Korean Medicine
/
v.22
no.3
/
pp.542-547
/
2008
In order to examine anti-inflammatory effect of Cinnamomi Ramulus (CR) alcohol extract on rheumatoid arthritis, the present study investigated the viability and TNF-${\alpha}$ production in Raw264.7 cells treated with CR and collagen induced arthritis in DBA/1J mice which were orally administered with CR prior to immunization. The results are as follows: CR extract at $20{\mu}g/ml$ inhibited the viability of Raw264.7 by 50%. CR at 15 and $20{\mu}g/ml$ showed a significant decrease in TNF-${\alpha}$ levels from Raw264.7 cells treated with LPS. CR administration decreased arthritic index in DBA/1J mice immunized with bovine collagen type II but it did not reach statistical significance. CR administration significantly decreased spleen weights obtained from mice in 6 weeks after immunization. CR administration significantly decreased serum anti-type II collagen antibody levels compared with control group. CR administration decreased serum IL-6 levels compared with control group but it did not reach statistical significance.
Kim Seok Kyun;Jung Sang Mi;Ahn Kyong Hoon;Jeon Hyung Jun;Lee Dong Hun;Jung Kwang Mook;Jung Sung Yun;Kim Dae Kyong
Archives of Pharmacal Research
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v.28
no.8
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pp.923-929
/
2005
Methanol extracts of domestic plants of Korea were evaluated as a potential inhibitor of neutral pH optimum and membrane-associated 60 kDa sphingomyelinase (N-SMase) activity. In this study, we partially purified N-SMase from bovine brain membranes using ammonium sulfate. It was purified approximately 163-fold by the sequential use of DE52, Butyl-Toyopearl, DEAE-Cellulose, and Phenyl-5PW column chromatographies. The purified N-SMase activity was assayed in the presence of the plant extracts of three hundreds species. Based on the in vitro assay, three plant extracts significantly inhibited the N-SMase activity in a time- and concentration-dependent manner. To further examine the inhibitory pattern, a Dixon plot was constructed for each of the plant extracts. The extracts of Abies nephrolepis, Acer tegmentosum, and Ginkgo biloba revealed a competitive inhibition with the inhibition constant (Ki) of $11.9 {\mu}g/mL,\;9.4{\mu}g/mL,\;and\;12.9{\mu}g/mL$, respectively. These extracts also inhibited in a dose-dependent manner the production of ceramide induced by serum deprivation in human neuroblastoma cell line SH-SY5Y.
Several experiments were carried for the purpose of establishing the basis for the quality evaluation of deer horn. Deer horn originated from China, New Zealand, Soviet and Alaska were used as objectives and some constituents involved in deer horn were assayed and compared with one another. Five kinds of trace elements including Ca and Fe were determined by atomic absorbance spectroscopy. The more the region came down from the top of the deer horn, the more the ratio of Ca to Fe content (Ca/Fe) was increased. The same increasing tendancy was observed in the case of ash content. Gangliosides were isolated from deer horn by alumina column chromatography and identified by TLC using gangliosides Type II obtained from bovine brain as the standard material. The TLC chromatograms were scanned by dual wavelength TLC scanner, then those TLC profiles were compared. Deer horn from China and New Zealand were analogous but those from Soviet and Alaska were each distinguished in their TLC profiles.
Rabies virus which belongs to the genus Lyssavirus of the family Rhabdoviridae is known as a highly neurotropic virus and causes fatal encephalitis accompanied by severe neurological symptoms in almost all mammals, including humans. In this study, monoclonal antibodies (MAbs) against rabies virus were produced, characterized and applications of MAbs as diagnostic reagents were assessed Spleen and inguinal lymph node cells from Balb/c mouse immunized with purified rabies virus were fused with SP2/O myeloma cells using polyethylene glycol (PEG) and hybridoma cells producing rabies virus-specific MAbs were screened by an indirect fluorescent antibody test. A total of ten MAbs were produced against rabies virus. The protein specificity and neutralizing activity of MAbs were determined by Western blot analysis and fluorescent antibody virus neutralization test, respectively. As a result, two MAbs, 5G3 and 6H4 had specificity for nucleoprotein (N protein) and two other MAbs, 5B1 and 5C1 had neutralizing activity for rabies virus. Some MAbs recognized the rabies virus-infected bovine brain stem cells by immunohistochemistry (IHC) assay. In conclusion, it was confirmed that MAbs produced in this study were rabies virusspecific and could be used as reliable diagnostic reagents for the detection of rabies virus.
Cerebral microvessel endothelial cells that form blood-brain barrier (BBB) have tight junction for maintaining brain homeostasis. Occludin, one of tight junction protein, is crucial for BBB function. $H_2O_2$ induced occludin changes and effects in bovine brain BBB endothelial cells were examined in this study. The decrease of transendothelial electrical resistance (TEER) by $H_2O_2$ was due to disruption of occludin localization. Cytotoxicity test revealed that $H_2O_2$ did not cause cell death below 1 mM $H_2O_2$ within 4 hr. $H_2O_2$ caused intermittent disruption and loss of occludin at tight junctions and occludin disappeared with dose dependent manner from tight junction in confocal laser microscopy. But Western blot revealed that the total amounts of occludin increased by $H_2O_2$ administration. Transmission electron microscopy revealed that the ultrastructure of tight junction was not changed by $H_2O_2$. These data suggest that functional disruption of BBB by $H_2O_2$ was due to the localized loss of occludin in tight junction, but the expression of occludin increased in order to compensate the disrupted function in BBB.
The transmissible spongiform encephalopathies (TSEs) disease group are fatal neurodegenerative disorders affecting a wide range of hosts. The group includes kuru and Creutzfeldt-Jakob disease (CJD) in humans, scrapie in sheep and goats and Bovine spongiform encephalopathy (BSE) in cattle. The exact nature of the infectious agent involved in the transmission of these diseases remains controversial. However, a central event in their pathogenesis is the accumulation in infected tissues of an abnormal form of a host-encoded protein, the prion protein (PrP). Whereas the normal cellular protein is fully sensitive to protease ($PrP^{sen}$), the disease-associated prion protein ($PrP^d$) is only partly degraded ($PrP^{res}$), its amino-terminal end being removed. BSE was first reported in the mid-80s in the UK. Ten years later, a new form of human prion disease, variant CJD (vCJD) developed in the wake of the BSE epidemic, and there is now strong scientific evidence that vCJD was initiated by the exposure of humans to BSE-infected tissues, thus indicating a zoonotic disease. However, the ban on the feeding of animal-derived proteins to ruminants, and the apparent lack of vertical transmission of BSE, have led to a decline in the incidence of the disease within cattle herd and therefore, an assumed decreased risk for human contacting vCJD. The origin of the original case(s) of BSE still remains an enigma even though three hypotheses have been raised. Hypotheses are i) sheep- or goat-derived scrapie-infected tissues included in meat and bone meal fed to cattle, ii) a previously undetected sporadic or genetic bovine TSE contaminating cattle feed or iii) originating from a human TSE through animal feed contaminated with human remains. A host cellular membrane protein ($PrP^C$), which is abundant in central nervous system tissue, appear to be conformationally altered in the diseased host into a prion protein ($PrP^{Sc}$). This $PrP^{Sc}$ is detergent insoluble and partially protease-resistant ($PrP^{res}$). The term $PrP^{res}$ is normally used to describe the protein detected after protease treatment, in techniques such as Western immunoblotting, and enzyme-linked immunosorbant assay using fresh/frozen tissue. Immunohistochemistry may performed with formalin-fixed tissues. Also, clinical signs of the BSE are one of the major diagnostic indicators. Recently, atypical forms (known as H- and L-type) of BSE have appeared in several European countries, Japan, Canada and the United States. An unusual case was also reported in a miniature zebu. The atypical BSE fall into two groups based on the relative molecular mass (Mm) of the unglycosylated $PrP^{res}$ band relative to that of classical BSE, one of the higher Mm (H-type) and the other lower (L-type). Both types have been detected worldwide as rare cases in older animals, at a low prevalence consistent with the possibility of sporadic forms of prion diseases in cattle. This raises the unwelcome possibility that vCJD could increase in the human population. Now, active surveillance program against BSE is going on in Korea. In regional veterinary service lab, ELISA is applied to screen the BSE in slaughter and confirmatory tests by Western immunoblotting and immunohistochemisty are carried out if there are positive or suspect in the screening test. Also, the ruminant feed ban is rigorously enforced. Removal of specified risk materials such as brain and spinal cord from cattle is mandatory process at slaughter to prevent the infected material from entering the human food chain.
In High-Intensity Focused Ultrasound (HIFU) treatment, effective localization of HIFU focus is important for developing a safe treatment plan. While Magnetic Resonance Imaging guided HIFU (MRIgHIFU) can visualize the ultrasound path during the treatment for localizing HIFU focus, it is challenging in ultrasound imaging guided HIFU (USIgHIFU). In the present study, a real-time ultrasound beam visualization technique capable of localizing HIFU focus is presented for USIgHIFU. In the proposed method, a short pulse, with the same center frequency of an imaging ultrasound transducer below the regulated acoustic intensity (i.e., Ispta < 720 mW/㎠), was transmitted through a HIFU transducer whereupon backscattered signals were received by the imaging transducer. To visualize the HIFU beam path, the backscattered signals underwent dynamic receive focusing and subsequent echo processing. From in vitro experiments with bovine serum albumin gel phantoms, the HIFU beam path was clearly depicted with low acoustic intensity (i.e., Ispta of 94.8 mW/㎠) and the HIFU focus was successfully localized before any damages were produced. This result indicates that the proposed ultrasound beam path visualization method can be used for localizing the HIFU focus in real time while minimizing unwanted tissue damage in USIgHIFU treatment.
Journal of Physiology & Pathology in Korean Medicine
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v.22
no.6
/
pp.1416-1422
/
2008
In order to examine anti-inflammatory effect of Anemarrhenae Rhizoma (AR) alcohol extract on rheumatoid arthritis, the present study investigated the viability and TNF-${\alpha}$ production in Raw264.7 cells treated with AR and collagen induced arthritis in DBA/1J mice which were orally administered with AR prior to immunization. The results are as follows: AR extract at 20 and 50${\mu}g$/ml inhibited the viability of Raw264.7 by 35% and 79%, respectively. AR showed a significant decrease in TNF-${\alpha}$ levels from Raw264.7 cells treated with LPS. AR administration significantly decreased arthritic index in DBA/1J mice immunized with bovine collagen type II. AR administration significantly decreased spleen weights obtained from mice in 6 weeks after immunization. AR administration significantly decreased serum anti-type II collagen antibody levels compared with control group. AR administration decreased serum IL-6 levels compared with control group but it did not reach statistical significance.
Long-chain polyunsaturated fatty acids (LC-PUFA) promote the development of brain and vision of the fetus, relieve inflammation, inhibit oral dysplasia of rumor cell, decrease the incidence of cardiovascular disease and regulate arrhythmia. ${\Delta}-6$ desaturase is the rate-limited enzyme in the desaturation process. This study reports the cloning, characterization and tissue expression of a ${\Delta}-6$ desaturase gene in the chicken. PCR primers were designed based on the predicted sequence of chicken ${\Delta}-6$ desaturase (accession number: XM421053) and used to isolate a cDNA fragment of 1,323 bp from chicken liver. Based on the 1,323 bp fragment an EST (BI390105) was obtained by BLAST. The EST and 5'nd of the 1,323 bp fragment were partially overlapped. Gene specific primers derived from the EST were used for amplification of the 5'nd. Another gene-specific primer derived from the 1,323 bp fragment was used for amplification of the 3'nd by 3'ACE. Then the three overlapping cDNA sequences obtained were assembled with DNAMAN software and a full-length ${\Delta}-6$ desaturase of 2,153 bp was obtained. The full-length cDNA contained an ORF of 1,335 bp with a 5'ntranslated region of 147 nucleotides followed by an ATG initiation codon. Stop codon TGA was at position 1,481-1,483 bp. The deduced amino acids shared an homology above 77% with bovine, mice, orangutan, rat and human. The protein sequence had three histidine-rich regions HDFGH (HisI region), HFQHH (HisII region) and HH (HisIII region), a cytochrome $b_{5}$-like domain containing a heme-binding motif and two transmembrane domains. Sequence analysis of the chicken genomic DNA revealed that the coding sequence of chicken ${\Delta}-6$ desaturase included 12 exons and 11 introns. Semi-quantitative RT-PCR showed that the ${\Delta}-6$ desaturase expression levels were in turn liver, spleen, pancreas, lung, breast muscle, heart, and abdominal fat. The expression of ${\Delta}-6$ desaturase in liver was significantly higher than that in breast muscle (p<0.01). The expression of ${\Delta}-6$ desaturase in lung was significantly higher than that in abdominal fat (p<0.01). This is the first clone of chicken ${\Delta}-6$ desaturase.
Telomeres consist of TTAGGG tandem repeated DNA sequences with specific proteins and locate at chromosome ends. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. Telomerase is a ribonucleoprotein which has a template for the synthesis of telomeric DNA. This study was carried out to analyze the amount of telomeric DNA and telomerase activity in cattle cells. Analysis of the quantity of telomere in lymphocytes was done at different ages, sex and among Korean cattle and Holstein breeds. The telomerase activity was also analyzed in liver, brain, heart, kidney, and testis tissues of fetal calf and of 18 month old cattle. The amount of telomeres in lymphocytes and other tissue cells was analyzed by Quantitative-Fluorescence in situ Hybridization (Q-FISH) technique using a telomeric DNA probe. Telomerase activity was analyzed by Telomeric Repeat Amplification Protocol assay (TRAP). The amount of telomeric DNA on the lymphocytes during the whole life span was decreased along with age. Quantity of telomeres in Korean cattle was significantly higher than that in Holstein breed. The amount of telomeric DNA in males was significantly higher than that in females. Telomerase activity was up-regulated in most bovine tissues during fetal stage, but was down-regulated in most tissues at mature 18 month age except the testis cells. This study indicates that the amount of telomeres and telomerase activity of cells can be used as an age marker or/and a physiological marker of cattle.
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