• Parasites, Hosts and Diseases
• /
• v.24 no.2
• /
• pp.177-186
• /
• 1986

This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of $2{\mu}g/ml$ antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: 1. Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at $51{\sim}65%$ precipitate (PA2), followed by $0{\sim}50%$ precipitate (PAl), $66{\sim}80%$ precipitate (PA3), and $81{\sim}90%$ precipitate (PA4). Unprecipitated protein over 90% ammonium sulfate (PA5) showed the lowest antigenicity. 2. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. 3. By passing through the Sephadex G-200 column, PA1 and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2-I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. 4. SDS-polyacrylamide gel electrophoresis of PA1-I, P A1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons(KD) to 196 KD; of them 220KD protein was major band. Fraction PA2-I was composed of $255{\sim}225\;KD$, and PA3-I, $255{\sim}240\;KD$, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. 5. Absorbance by ELISA began to increase within $10{\sim}20$ days after the infection and reached the highest on $140{\sim}180$ days, then made plateau thereafter. 6. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. 7. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at $20{\sim}30$ days after infection.

• Parasites, Hosts and Diseases
• /
• v.30 no.4
• /
• pp.309-322
• /
• 1992

In order to observe the antigenic localization in the tissues of Metngonimus yokogawai in growth stages, immunogoldlabeling method was applied to using serum of the cat which Infected with isolated metacercariae from Plecoglossus aztivelis. The sectioned worm tissues from each growth stages were embedded in Lowicryl HM 20 medium, stained with infected serum IgG and protein A gold complect (particle size: 12 nm) and observed by electron microscopy. In the worm tissues of all experimental groups, the geld particles were specifically concentrated on the tegumental synch- tium and cytoplasm of the tegumental cell as well as the secretory granules in the parenchymal tissue. In the 16th and 20th week grown worm tissues, the gold particles were specifically concentrated on the vesicles in the tegumental syncytium and cl·toplasm of the tegumental cell. The gold particles were specifically concentrated on the caecal epithelia of the 4th, 8th and 12th week growth groups but slightly concentrated on those of the 16th and 20th week.

• Journal of Life Science
• /
• v.9 no.4
• /
• pp.439-452
• /
• 1999

This study was designed to investigate the effects of sea tangle (Laminaria japonica) extract and fucoidan components on anti-aging action. Sprague-Dawley(SD) male rats (210$\pm$5g) were fed experimental diets Dasi-Ex group: sea tangle extract powder of 4.0% added to control diet; Fuco-I, II and III groups: funcoidan powder of 1, 2 and 3% added to Dasi-Ex group for 45 days. Hydroxyl radical (.OH) formations were significantly inhibited (10-20% and 25-30%) in serum and brain mitochondria of Dasi-Ex and Fuco-I, II and III groups compared with control group. Significant differences in .OH formations of brain mitochondria in Dasi-Ex and Fuco-I groups could not be obtained, but.OH formations of brain microsomes resulted in a significant decrease (15-20%) in Fuco-II and III groups compared with control group. Basal oxygen radical (BOR) formations were significantly decreased about 10% and 13-15% in brain mitochondria of Dasi-Ex and Fuco-I group, and Fuco-II, III groups, and also decreased about 10% and 15-20% in brain microsomes of Dasi-Ex and Fuco-I groups, and Fuco-II, III groups. LPO levels of brain mitochondria and microsomes were significantly inhibited about 10% in Dasi-Ex and Fuco-I, II groups and 15% in Fuco-III groups. Oxidized proteins (>C=O) were significantly inhibited about 10% in serum of Dasi-Ex and Fuco-I, II, III groups and brain mitochondria of Dasi-Ex group, while remarkably inhibited (30~35%) in brain mitochondria of Fuco-I, II and III groups. Nitric oxide (NO) levels were significantly inhibited (12~15%) in serum of Fuco-I, II and III groups, but there no significant difference in serum NO levels of Dasi-Ex group. Superoxide dismutase (SOD) activities were remarkably increased (30~ 60%) in serum of Fuco-I, II and III groups, but there were no significant differences in SOD activities in serum of Dasi-Ex group. Catalase (CAT) activities were significantly increased about 20% in serum of Dasi-Ex and Fuco-I, II, III groups. Mn-SOD activities in brain mitochondria were significantly increased about 17% in Dasi-Ex group, while remarkably increased 26~36% in Fuco-I, II, III groups. Cu,Zn-SOD activities in brain cytosol were dose-dependently of fucoidan increased 10%, 12% and 18%, respectively, compared with control group. These results suggest that anti-aging effects of fucoidan may play a pivotal role in attenuating a various age-related changes such as chronic degenerative disease and senile dementia.

• Advances in Traditional Medicine
• /
• v.8 no.2
• /
• pp.154-163
• /
• 2008

The methanol extract of stem barks of Careya arborea Roxb. (MECA) (Family- Myrtaceae) was evaluated for antitumor activity and antioxidant status against Ehrlich's Ascites Carcinoma (EAC) bearing Swiss albino mice. After 24 h of tumor inoculation the MECA was administered at the doses of 50, 100 and 200 mg/kg body weight/mice/day for 14 days. After the last dose and 18 h fasting mice were sacrificed. The effect of MECA on the growth of transplantable murine tumor, life span of EAC bearing hosts, hematological profiles, serum and liver biochemical parameters were estimated. The MECA showed significant (P < 0.01) decrease in ascites volume, packed cell volume and viable cell count and prolonged the life span of EAC tumor bearing mice. Hematological profiles reverted to more or less normal levels in extract treated mice. The MECA also produced protective effect by decreasing the activity of serum enzymes, bilirubin and increase the protein and uric acid levels. MECA significantly (P < 0.05) decreased the levels of lipid peroxidation, while significantly (P < 0.05) increased the levels of glutathione content, vitamin C, vitamin E, superoxide dismutase and catalase CAT. The results indicate that MECA exhibited significant antitumor and antioxidant activity in EAC bearing mice.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.3
• /
• pp.518-524
• /
• 2010

A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and $60^{\circ}C$, respectively. The activity of Man26B was not affected by $Mg^{2+}$ and $Co^{2+}$, but was inhibited by $Hg^{2+},\;Ca^{2+},\;Cu^{2+},\;Mn^{2+},\;K^+,\;Na^+$, and $\beta$-mercaptoethanol, and slightly enhanced by $Pb^{2+}$ and $Zn^{2+}$. EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with $K_m,\;V_{max}$, and $k_{cat}$ values of 3.80 mg/ml, 91.70 ${\mu}mol$/min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at $80^{\circ}C$ and $90^{\circ}C$ for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.

• Korean Journal of Microbiology
• /
• v.51 no.1
• /
• pp.68-74
• /
• 2015

Based on the genomic analysis of Thermococcus pacificus P-4, we identified a putative GH1 ${\beta}$-glucosidase-encoding gene (Tpa-glu). The gene revealed a 1,464 bp encoding 487 amino acid residues, and the deduced amino acid residues exhibited 77% identity with Pyrococcus furiosus ${\beta}$-glucosidase (accession no. NP_577802). The gene was cloned and expressed in Escherichia coli system. The recombinant protein was purified by metal affinity chromatography and characterized. Tpa-Glu showed optimum activity at pH 7.5 and $75^{\circ}C$, and thermostability with a half life of 6 h at $90^{\circ}C$. Tpa-Glu exhibited hydrolyzing activity against various pNP-glycopyranosides, with kcat/Km values in the order of pNP-${\beta}$-glucopyranoside, pNP-${\beta}$-galactopyranoside, pNP-${\beta}$-mannopyranoside, and pNP-${\beta}$-xylopyranoside. In addition, the enzyme exhibited exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharide (laminarin) and ${\beta}$-1,3- and ${\beta}$-1,4-linked oligosaccharides. This is the first description of an enzyme from hyperthermophilic archaea that displays exo-hydrolyzing activity toward ${\beta}$-1,3-linked polysaccharides and could be applied in combination with ${\beta}$-1,3-endoglucanase for saccharification of laminarin.

• YAKHAK HOEJI
• /
• v.56 no.1
• /
• pp.9-13
• /
• 2012

We previously reported that the extract of Taraxacum platycarpum (AF-343) had several biological properties such as skin hydration and anti-inflammatory effects, thereby AF-343 be a promising anti-atopic dermatitis agent. However, few studies have been conducted to evaluate its effect on modulation of extracellular matrix proteins in human skin fibroblasts. The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in AF-343-treated human skin fibroblasts. Human skin fibroblasts were treated by various concentrations of AF-343 (0~2 mg/ml). The expressions of type I collagen, matrix metalloproteinase-1 (MMP-1), Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, level of type I collagen mRNA was analyzed by CAT assay. Expression of type I collagen protein was increased in AF-343-treated human skin fibroblasts by dose and time-dependent manners. Consistent with this result, the expressions of phospho-Smad2/3 in skin fibroblasts were increased and MMP-1 expression was decreased by AF-343 treatment. TIMP-1 expression was not significantly changed in AF-343 treated skin fibroblasts. Extract of Taraxacum platycarpum (AF-343)-induced up-regulation of type I collagen expression was through increased expression of phospho-Smad2/3. These results were occurred combined with down-regulation of MMP-1 in skin fibroblasts. Taken together, this study indicated that AF-343 has property of the modulation of ECM in tissue as well as skin hydration and anti-inflammation.

• Journal of Veterinary Clinics
• /
• v.24 no.2
• /
• pp.104-108
• /
• 2007

The purpose of the study is to evaluate the clinical antagonistic effect of atipamezole(0.25 mg/kg, IM) in cats anesthetized with tiletamine-zolazepam ($Zoletil^{(R)}$, 10 mg/kg, IM) and medetomidine (0.05 mg/kg, IM). Twelve healthy 1 year old Korean mixed breed cats were used for this study. They were 4 males and 8 females. These cats were randomly assigned to two groups. One was control group ($Zoletil^{(R)}$ + medetomidine, ZM), and the other was treatment group ($Zoletil^{(R)}$ + medetomidine and antagonism by atipamezole, ZMA). All cats were examined 15 minutes before, 5, 25, 65 and 105 minutes after administration of tiletamine-zolazepam and medetomidine. Atipamezole was injected intramuscularly 20 minutes after ZM administation. Recovery time, heart rate, respiratory rate, total plasma protein and blood glucose were significantly different between ZM group and ZMA group (P<0.05). However, rectal temperature was not significantly different between ZM group and ZMA group. Two groups were able to induce sternal recumbency within 2 minutes and lateral recumbency within 4 minutes after the anesthetics injection. Mean sternal position time ($mean{\pm}SD$) was $174.0{\pm}44.6\;and\;116.2{\pm}27.3$ minutes, and mean standing position time was $210.8{\pm}45.6\;and\;154.2{\pm}21.1$ minutes in ZM and ZMA group, respectively. In these two groups, adverse effects during recovery time from anesthesia were not seen. As a result, the ZMA group had a faster recovery than the ZM group. Thus it was concluded that atipamezole could exert a useful reversal effect in cats anesthetized with medetomidine-tiletamine/zolazepam combination.

• Archives of Pharmacal Research
• /
• v.24 no.6
• /
• pp.546-551
• /
• 2001

We have previously shown that, in circular muscle cells of the lower esophageal sphincter (LES) isolated by enzymatic digestion, contraction in response to maximally effective doses of acetylcholine (ACh) or Inositol Triphosphate ($IP_3$) depends on the release of $Ca^{2+}$ from intracellular stores and activation of a $Ca6{2+}$-calmodulin (CaM)-dependent pathway. On the contrary, maintenance of LES tone, and response to low doses of ACh or $IP_3$ depend on a protein kinase C (PKC) mediated pathway. In the present investigation, we have examined requirements for $Ca6{2+}$ regulation of the interaction between CaM- and PKC-dependent pathways in LES contraction. Thapsigargin (TG) treatment for 30 min dose dependently reduced ACh-induced contraction of permeable LES cells in free $Ca6{2+}$ medium. ACh-induced contraction following the low level of reduction of $Ca6{2+}$ stores by a low dose of TG ($10^{-9}{\;}M$) was blocked by the CaM antagonist, CCS9343B but not by the PKC antagonists chelerythrine or H7, indicating that the contraction is CaM-dependent. After maximal reduction in intracellular $Ca{2+}$ from $Ca6{2+}$stores by TG ($10^{-6}{\;}M$), ACh-induced contraction was blocked by chelerythrine or H7, but not by CCS9343B, indicating that it is PKC-dependent. In normal $Ca^{2+}$medium, the contraction by ACh after TG ($10^{-9}{\;}M$) treatment was also CaM-dependent, whereas the contraction by ACh after TG ($10^{-9}{\;}M$) treatment was PKC-dependent. We examined whether PKC activation was inhibited by activated CaM. CCS 7343B Inhibited the CaM-induced contraction, but did not inhibit the DAC-induced contraction. CaM inhibited the DAC-induced contraction in the presence of CCS 9343B. This inhibition by CaM was $Ca{2+}$dependent. These data are consistent with the view that the switch from a PKC-dependent pathway to a CaM dependent pathway can occur and can be regulated by cytosolic $Ca{2+}$ in the LES.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.48 no.2
• /
• pp.123-134
• /
• 2022

In this study, the whitening and antioxidant effects of the extracts from Hydrangea petiolaris (H. petiolaris) leaves was confirmed, and the chemical structure was identified by separating the active ingredients. In the whitening tests using α-MSH stimulated B16F10 melanoma cells, the n-hexane (Hex) fraction inhibited the cellular melanogenesis and intracellular tyrosinase activities without causing cell toxicity. In addition, the Hex fraction reduced expression of tyrosinase and TRP-2 protein. Upon the anti-oxidative studies by DPPH and ABTS⁺ radicals, potent radical scavenging activities were observed in the ethyl acetate (EtOAc) fraction. Also, for the cellular protective effects on HaCat keratinocytes damaged by H₂O₂, the EtOAc fraction indicated protective effects against oxidative stress. Eight phytochemicals were isolated from the extract of H. petiolaris leaves; ethyl linoleate (1), ethyl linolenate (2), 1-linoleoyl glycerol (3), 1-linolenoyl glycerol (4), epi-catechin (5), afzelin (6), quercitrin (7), hyperin (8). Among the isolates, the compounds 5 - 8 showed DPPH and ABTS⁺ radical scavenging activities. The contents of quercitirin, a major isolated in this extract, determined by HPLC analysis were confirmed to be about 31.3 mg/g for the 70% ethanol extract and 169.8 mg/g for the EtOAc fraction. Based on these results, it was suggested that the extract from H. petiolaris leaves could be potentially applicable as whitening and anti-oxidative ingredients in cosmetic formulations.