Deer antler has been widely prescribed in Chinese and Korean pharmacology. Although there have been several reports concerning the effects of deer antler, such as anti-aging action, anti-inflammatory activity, antifungal action and regulatory activity of the level of glucose, the effect on bone has not determined yet. The purpose of this study was to examine the effect of deer antler on osteoblast differentiation. Hexane extract(CN-H) and chloroform extract(CN-C) were acquired from deer antler(Cervus nippon) and MC3T3-E1 pre-osteoblasts were cultured in the presence or absence of each extract. Osteoblast differentiation was estimated with the formation of mineralized nodules and the mRNA expression of alkaline phosphatase(ALP), osteocalcin(OC) and bone sialoprotein(BSP) which are markers of osteoblast differentiation. Non-treated group did not show mineralized nodule. CN-C or CN-H-treated group showed minerlaized nodules in 16 days. In northern blot analysis, CN-C or CN-H-treated group showed the elevated expression of ALP, BSP and OC in 16 days. These results suggest the possibility to develop deer antler as a bone regenerative agent in periodontal therapy by showing the stimulating activity of deer antler on differentiation of osteoblast.
Ji, Hye-Young;Lee, Seung-Seok;Yoo, Sung-Eun;Kim, Hosoon;Lee, Dong-Ha;Lim, Hong;Lee, Hye-Suk
Archives of Pharmacal Research
/
v.27
no.2
/
pp.239-245
/
2004
KR-31543, (2S,3R,4S)-6-amino-4-[N-(4-chlorophenyl)-N-(2 -methyl-2H-tetrazol-5-ylmethyl) amino]-3,4-dihydro-2-dimethoxymethyl-3-hydroxy-2-methyl-2H-1-benzopyran, is a new neuroprotective agent for preventing ischemia-reperfusion damage. This study was performed to identify the metabolic pathway of KR-31543 in human liver microsomes and to characterize cytochrome P450 (CYP) enzymes that are involved in the metabolism of KR-31543. Human liver microsomal incubation of KR-31543 in the presence of NADPH resulted in the formation of two metabolites, M1 and M2. M1 was identified as N-(4-chlorophenyl)-N-(2-methyl-2H-tetrazol-5-ylmethyl)amine on the basis of LC/MS/MS analysis with a synthesized authentic standard, and M2 was suggested to be hydroxy-KR-31543. Correlation analysis between the known CYP enzyme activities and the rates of the formation of M 1 and M2 in the 12 human liver microsomes have showed significant correlations with testosterone 6$\beta$-hydroxylase activity (a marker of CYP3A4). Ketoconazole, a selective inhibitor of CYP3A4, and anti-CYP3A4 monoclonal antibodies potently inhibited both N-hydrolysis and hydroxylation of KR-31543 in human liver microsomes. These results provide evidence that CYP3A4 is the major isozyme responsible for the metabolism of KR-31543 to M1 and M2.
Biphenyl dimethyl dicarboxylate (DDB) is a hepatoprotectant, which is used as an adjuvant agent in a treatment for chronic hepatitis. Amantadine is an antiviral agent, which is utilized primarily in the treatment of influenza, but also, occasionally in the treatment of hepatitis C. In a previous study, we reported that DDB, coupled with amantadine, would exert an anti-HBV effect, via the induction of interferon-inducible gene expression in the HepG2 2.2.15 cell line. The primary objective of the present study was to determine whether or not DDB and/or amantadine exhibit anti-HBV properties, and what mechanisms of action might be involved in such properties. In our study, we were able to determine that DDB stimulates Jak/Stat signaling, and induces the expression of interferon alpha $(IFN-\alpha)$ stimulated genes, most notably 6-16 and ISG12. In addition, the antiviral effectors induced by $IFN-\alpha$, PKR, OAS, and MxA, were regulated in the presence of DDB at its optimal concentration $(250{\mu}g/mL)$, to a degree commensurate with the degree of induction associated with the $IFN-\alpha$ treated group. Finally, we determined that the replication of pregenomic RNA and HBeAg was inhibited by DDB treatment, and this inhibition was maximized when coupled with the administration of amantadine $(25{\mu}g/mL)$. In conclusion, the results of this study demonstrated clearly that DDB, as well as the combination of DDB/amantadine, directly inhibited $IFN-\alpha$ signaling-mediated replication of HBV in infected hepatocytes, and thus may represent a novel treatment for chronic hepatitis B, which would be characterized principally by its improved safety over other treatment strategies.
Yi, Su Hee;Lee, Jin Heung;Seo, Sang Tae;Lee, Jong Kyu
Journal of Forest and Environmental Science
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v.33
no.4
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pp.342-347
/
2017
Since the first report of the oak wilt disease at 2004 in Korea, the disease distributed over Korean peninsula and are still giving severe damages. The management of oak wilt disease in Korea has mainly focused on the control of insect vector, Platypus koryoensis. Neverthless the effective method for evaluating the pathogenicity of the pathogen, Raffaelea quercus-mongolicae (Rqm), and for screening chemical or biological agents with strong inhibitory activity against the pathogen, is absolutely necessary, an reliable method is not available so far. This study was conducted to develop the effective method for evaluating the pathogenicity of Rqm in oak trees. The culture suspensions of Rqm were artificially injected to the saplings of Quercus acutissima by using ChemJet tree injector. Three months after treatments, the treated saplings were cut and dipped into 1% fuchsin acid solution. There were significant differences in non-conductive area (%), discoloration area (%) and vertical discoloration length between the pathogen-injected and distilled water-injected control treatments. These results indicated that the pathogen is the causal agent for the dysfunction of water conductive tissue, which will finally result in wilt symptom. Re-isolation of the pathogen and PCR detection using specific primers for the pathogen also confirmed the presence of Rqm in the sapwood chips of the pathogen-injected saplings. These observations would be greatly applied to other related researches for evaluating the pathogenicity of tree wilt pathogens and biocontrol efficacy of the selected antagonistic microorganisms, in case that the wilt symptom is not easily shown by artificial inoculation of the causal agent.
The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.
JeongWoo Kang;Ah-Young Kim;Hyun Young Chae;Hanae Lim;Suncheun Kim;Bok-Kyung Ku;Kyunghyun Lee
Korean Journal of Veterinary Research
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v.63
no.3
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pp.21.1-21.6
/
2023
The increasing prevalence of toxic substance-exposure in pets in South Korea endangers the health and safety of numerous companion animals, and has become a cause for concern. Notably, the annual incidence of forensic analysis in pets has increased by more than 150% in South Korea, mainly in populous regions such as Seoul, Incheon, and Gyeonggi. In response to this growing issue, veterinary forensic examinations were conducted on 549 dogs and cats from 2019 to 2022. This study revealed the presence of various toxic substances, including pesticides, insecticides, and drugs such as analgesics, anesthetics, antidepressants, and muscle relaxants, in pets. Among the 38 different toxins identified in pets, coumatetralyl, methomyl, terbufos, and buprofezin were the most frequently detected. In this study, toxic substances for pets were identified based on the "toxic agent list for humans," developed by the National Forensic Services, because no list of toxic agents for animals currently exists and data regarding potentially toxic substances for dogs and cats is limited. This is one of the limitations of this study, and necessitates the establishment of a toxic agent list for animals. Continued monitoring and research is also recommended to reveal the incidence, causes, and solutions of toxicity in animals.
Antioxidant activity of 70% aqueous ethanolic extract of leaves of Justicia gendarussa (EJ) was evaluated. EJ was prepared by cold maceration method. The antioxidant potency of EJ was investigated employing various established in vitro systems, such as DPPH radical scavenging, nitric oxide (NO) scavenging, ${\beta}-carotene$ linoleic acid module system (${\beta}$ CLAMS), hydroxyl (OH) radical scavenging, anti lipid peroxidation. $IC_{50}$ values were determined in each experiment. Also, ferric ion reduction capacity of extracts in presence and absence of chelating agent (EDTA) and total antioxidant capacity were determined. Preliminary phytochemical investigation was carried out to know the nature of constituents present in the leaves and correlate it with antioxidant activity. Further total phenolic content was determined in EJ. $IC_{50}$ values of EJ were 123.09 ${\pm}$ 3.01, 643.0 ${\pm}$ 61.10, 132.3 ${\pm}$ 6.03, 68.5 ${\pm}$ 11.5 and 68.13 ${\pm}$ 1.38 ${\mu}g/mL$ in DPPH radical scavenging, NO scavenging, ${\beta}$ CLAMS, OH radical scavenging and anti lipid peroxidation activity respectively. In total antioxidant capacity assay, ascorbic acid equivalent value was found to be 205.56 ${\pm}$ 4.69 ${\mu}g/mg$ of extract. Total phenolic content was found to be 43.76 ${\pm}$ 4.27 ${\mu}g$ equivalent of gallic acid per mg of extract. Phytochemical investigation reveals the presence of flavonoids. The results indicate that EJ possess antioxidant activity and flavonoids are responsible for this activity.
A number of investigations have shown that the presence of bacteria is prerequisite for developing pulpal and/or periradicular pathosis. Depending on the stage of pulpal pathosis, various species of bacteria can be cultured from infected root canals. Kakehashi et al. showed that exposure of pulpal tissue in germ-free rats was characterized by minimal inflammation and dentinal bridging while exposure of pulpal tissue in conventional rats with normal oral flora was characterized by pulpal necrosis, chronic inflammation, and periapical lesions. Currently used methods of cleaning and shaping, especially rotary instrumentation techniques, produce a smear layer that covers root canal walls and the openings to the dentinal tubules. The smear layer contains inorganic and organic substances that include fragments of odontoblastic processes, microorganisms, their by products and necrotic materials. Because of its potential contamination and adverse effect on the outcome of root canal therapy, it seems reasonable to suggest removal of the smear layer for disinfection of the entire root canal system. Presence of this smear layer prevents penetration of intracanal medications into the irregularities of the root canal system and the dentinal tubules and also prevents complete adaptation of obturation materials to the prepared root canal surfaces. Removal of the smear layer by an intracanal irrigant and placement of an antibacterial agent in direct contact with the content of dentinal tubules should allow disinfection of this complex system and better outcome for the root canal therapy. A new solution, which was a mixture of a tetracycline, an acid, and a detergent(MTAD), was developed in the Department of Endodontics, Dental School. Lorna Linda University, USA. It has been demonstrated that MTAD was an effective solution for the removal of the smear layer and does not significantly change the structure of the dentinal tubules when used as a final irrigant in conjunction with 1 % NaOCl as a root canal irrigant. Studies are in progress to compare the anti- microbial properties of this newly developed solution with those of sodium hypochlorite and EDTA that are currently used to irrigate the root canals and remove the smear layer from the surfaces of instrumented root canals.canals.
The present study was conducted to investigate the effects of bornyl acetate on arterial blood pressure and vascular contractile responses in the normotensive rats and to establish the mechanism of action. Both phenylephrine (an adrenergi$\alpha$-receptor agonist) and high potassium (a membrane-depolarizing agent) caused greatly contractile responses in the isolated aortic strips. These phenylephrine (10$^{-5}$ M)-induced contractile responses were depressed in the presence of high concentrations of bornyl acetate (10∼20 $\mu\textrm{g}$/ml), but not affected in low concentrations of bornyl acetate (2.5∼5$\mu\textrm{g}$/ml). High potassium (5.6 ${\times}$ 10$^{-2}$ M)-induced contractile responses were also greatly inhibited in the presence of bornyl acetate (2.5∼20 $\mu\textrm{g}$/ml) in a dose-dependent fashion. Bornyl acetate (1∼10 mg/kg) given into a femoral vein of the normotensive rat produced a dose-dependent depressor response, which is transient (data not shown). Interestingly, the infusion of a moderate dose of bornyl acetate (3mg/kg/30 min) made a significant reduction in pressor responses induced by intravenous norepinephrine. Collectively, these results obtained from the present study demonstrate that intravenous bornyl acetate causes a dose-dependent depressor action in the anesthetized rat at least partly through the blockade of adrenergic $\alpha$$_1$-receptors. bornyl acetate also causes vascular relaxation in the isolated aortic strips of the rat via the blockade of adrenergic $\alpha$$_1$-receptors, in addition to the unknown mechanism of direct vasorelaxation.
Since 1993, the White Spot Syndrome Virus (WSSV) disease occurred in China among cultured shrimps resulting in mass mortality. Epizootiological surveys undertaken during the outbreak period of 1993-1994 indicated that all stages of Penaeus chinensis, P. japonicus and P. monodon were infected. Consequent to the transport of contaminated shrimp seedlings and seawater, the disease spread all over the farms of China. The disease was more rapidly transmitted at temperatures above $25^{\circ}C$. Challenge experiments showed the causative agent was highly virulent. White spots appeared on the carapace of both span-taneous and experimentally infected shrimps. Moribund shrimps contained turbid hemolymph, hypertrophied Iymphoid organ and a necrotic mid-gut gland. Electron microscopy showed the presence of viral particles in the gills, stomach, lymphoid organ, and epidermal tissue of the infected shrimp. The visions were slightly ovoid with an envelope and averaged 350 $\times$ 150 nm; nucleocapsids measured 375 $\times$ 157 nm. With discontinuous sucrose gradient of 35, 50 and 60% (w/v), the virus was separated from hemolymph of the infected shrimp. The estimated molecular weight of genomic DNA was 237 Kb with EcoR I, 247 Kb with Hind III and 241kb with Pst I. A total of 9 hybridoma colones secreting monoclonal antibodies (MAbs) were produced from mouse myeloma and spleen cells immunized with WSSV. The immunofluorescence assay of gill tissue showed that the MAbs reacted with diseased but not with healthy shrimp. The MAbs belonged to IgGl, IgG2b subclass and IgM class, all with kappa light Immune-electron-microscopy with colloidal gold marker showed the presence of 5 MAbs epitopes on the envelope and one on the capsid of the virus. Baculoviral mid-gut gland necrosis showed the specificity of the MAbs produced. For diagnosis 5 different methods were selected. Using Kimura primers for PCR, or MAbs for immunoblot, ELISA or FAT method, in situ hybridization was carried out to show the gene. All these methods detected WSSV in the organ samples of the diseased shrimp but not in healthy one.
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