Clonorchis sinensis is a common parasite of man in Korea. Researches on the specific antigens of C. sinensis would be valuable not only because those elucidate the molecular characteristics of this fluke but also because it is applicable to immunodiagnosis. Although many monoclonal antibodies have been used in the field of parasite immunology, few articles on monoclonal antibodies against C. sinensis have been published so far. The aim of this study was to analyse C. sinensis antigens recognized by monoclonal antibodies, and to set up ELISA-inhibition test using C. sinensis specific monoclonal antibodies for improved specificity of immunodiagnostic tests. By fusion between spleen cells of the mice immunized with C. sinensis water-soluble crude adult worm antigens and plasmacytoma cells of mouse origin, 29 hybridoma clones secreting anti-C. sinensis monoclonal antibodies were made, and 8 clones among those were found specific. After cell cloning, isotypes of 6 selected specific monoclonal anti- bodies were determined to be IgGl, IgG2b and IgA. Four exposed antigenic determinants of natural infection were recognized by different specific monoclonal antibodies. By enzyme-immunoelectrotransfer blot, 10 KD, 34 KD antigenic determinants were found to be reacted with CsHyb 0714-20, CsHyb 0605-10 monoclonal antibodies, respectively, The antigenic determinant recognized by CsHyb 0714-20 monoclonal antibody was revealed to be located at the surface and parenchyme of a parasite by indirect immunoauorescent antibody technique, and those reacted with CsHyb 0605-10, CsHyb 0714-25 monoclonal antibodies were found at the parenchyme and intestine. The antigenic determinant reacted with CsHyb 0605-23 monoclonal antibody was found mainly around the uterine eggs. Four antigenic determinants recognized by specific monoclonal antibodies were all found to be present in the early eluted fractions of C. sinensis antigens separated by Sephadex G-200 gel filtration. By conventional ELISA, 75% of clonorchiasis cases were found positive, but 7.1% of normal controls and 37.5% of paragonimiasis cases showed false positives. However, by ELISA-inhibition test using C. sinensis specific monoclonal antibody (CsHyb 0605-23), 77.1% of clonorchiasis cases were found positive, and there were no false positives in normal controls or paragonimiasis cases, indicating 100% specificity. The ELISA- inhibition test using monoclonal antibodies was found to have same sensitivity and definitely high specificity in comparison with conventional ELISA for serodiagnosis of human clonorchiasis.
Kim, Yeon-Ho;Cha, Sung-Ho;Ma, Sang-Hyuk;Kim, Ki-Sang;Lee, Young-Hee
Pediatric Infection and Vaccine
/
v.9
no.1
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pp.79-84
/
2002
Purpose : About 41% of obtained group A streptococci in the 1998 was reported as erythromycin-resistant streptococci in Seoul, Korea. The most common T serotype was T12, followed by T4 and T28. We'd like to monitor the serological changes and antibiotic sensitivity test of Streptococcus pyogenes obtained from the patients with pharyngotonsillitis and invasive diseases from 1999 through 2001. Also, it could be proposed to choose the proper antibiotic selection in the area where the rate of erythromycin-resistant streptococci is high. Methods : From Jan. 1999 to Oct. 2001, 208 isolates of group A streptococci were collected from inpatients and outpatients with pharyngotonsillitis, scarlet fever, and invasive infections in Seoul and Southern part of peninsula. All isolates were serotyped by T-agglutination, minimum inhibitory concentrations(MICs) which were determined by agar dilution methods, according to the guidelines of the National Committee for Clinical Laboratory Standards (NCCLS). Results : The most common T serotype was T12(29.8%), followed by T1(23.1%), T4 (14.9%). T1 was prominent serotype compared with previous year. T serotyping, among 25 isolates obtained from the patients with scarlet fever in Southern part of peninsula mostly, was T12, T1, and T4 in order of frequency. All the isolates tested were susceptible to penicillin, cefprozil, vancomycin, ceftriaxone, and chloramphenicol. However, 23 isolates(14.2%) was resistant to erythromycin and 18 isolates(11.1%) was resistant to clarithromycin. Serotype T12 was found to be the most resistant serotype to erythromycin and/or clarithromycin. Conclusion : High rate of erythromycin-resistant streptococci which surveyed in 1998 were reduced to 14.2% in this study. We should have to further evaluate the reason of decreased resistant strains and consider the resistant strains of streptococci in choosing the antibiotics. There was no serological characteristics according to the types of disease entities. Between the serologic distributions in Seoul and the Southern part of peninsula area are same, we could presume that the serological typing of strains obtained over the country may be not different.
In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.
The effect of low pH on physiological changes was studied with the acidophilic green alga, Clamydomonas acidophila, UTCC 122. The growthrates (${\mu}$) were identical, $0.5{\sim}0.7\;day^{-1}$, at pH 3.7${\sim}$6.7 and no significantly different (ANOVA, p =0.134), showing cell volume reduced gradually as they were growing, whereas that at pH 2.7 was falling to zero and cell volume increased dramatically. Chlorophyll a concentration of the cultures incubated for one day was $191{\sim}255\;pg\;cell^{-1}$, after then it declined from $60{\sim}103\;pg\;cell^{-1}$ at pH 3.7${\sim}$6.7 except $210\;pg\;cell^{-1}$ at pH 2.7, which was directly related with cell volume. External carbonic anhydrase (CA) activity was varied from1.1 to$3.7{\times}10^{-4}\;E.U.\;mm^{-2}$, showing the gradualincrease during culture, except at 2.7 and pH 5.7. However there was not found any relationship among the pH gradient cultures. CA molecular mass of C. acidophila was 29 kBa, and concentration of that was identical in all cultures. The proteins of 41 kDa and 63 were not or very faintly expressed in low pH cultures, in contrast that of 17 kDa more expressed. In this work, we found that C. acidophila could live optimally within a wide range of acidic pH, and 17 kDa of unidentified protein might be concerned with tolerating in low acid environment.
Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
Horticultural Science & Technology
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v.33
no.1
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pp.70-82
/
2015
This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.
We analyzed content and peptides in order to investigate the productivity from Cheonggookjang(fast-fermented soybean paste), fermenting it for 180 hours at $40^{\circ}C$. Results showed that pH was 7.07 at the start and became 7.41 in 24 hours, it eventually increased to 8.63 after 180 hours. Acidity was 0.2 in 12 hours, 0.5 in 12 hours, and then remained on 0.1 thereafter. Total sugar was 1.54 mg/$m\ell$ at the start, but it gradually decreased to 0.76 mg/$m\ell$ after the lapse of 48 hours, and 1.0 mg/$m\ell$ in 120 hours, and finally 0.8 mg/$m\ell$ in 180 hours. Reducing sugar was 0.14 mg/$m\ell$ at the start, and 0.88 mg/$m\ell$ after the lapse of 24 hours, 0.64 mg/$m\ell$ in 48 hours, 0.26 mg/$m\ell$ in 72 hours, and showed no definite change untill 180 hours. The amount of free amino acid was $0.19\;{\mu}M/\ell$ at the start, and $4.88\;{\mu}M/\ell$ after the lapse of 72 hours, $4.5\;{\mu}M/\ell$ in 120 hours, and then it rapidly decreased to $0.23\;{\mu}M/\ell$ after180 hours. Absorbance of soluble protein and peptide at 280 nm was 12.4 in 48 hours, 31.12 in 120 hours, and 31.12 in 180 hours. HPLC revealed that in the fermentation process, large molecular proteins are hydrolyzed into small peptides and amino acids, and after the lapse of 48 hours the pattern became almost the same. The protease activity of Cheonggookjang was 0.011 unit/$m\ell$ after the lapse of 36 hours and then it decreased. The result shows as Cheonggookjang started its deamination of amino acid in 100 hours, it is desirable to produce peptide within 100 hours of its fermentation.
Molecular relaxation behaviors of crystalline glucose anhydrous, crystalline glucose monohydrate, and amorphous glucose with different particle sizes were observed by measuring spin-spin relaxation time constant $(T_2)$ at the temperature range of $-20\;to\;110^{\circ}C$ using temperature-controlled low field nuclear magnetic resonance spectroscopy. No change in $T_2$ values of crystalline glucose anhydrous was observed throughout the temperature range, whereas $T_2$ values of crystalline glucose monohydrate and amorphous glucose increased from around $45\;and\;65^{\circ}C$, respectively. These results indicate that molecular mobility of crystalline glucose anhydrous does not change even at temperature higher than $100^{\circ}C$ and that the stability of powdered glucose could be improved by increasing the particle size of materials.
Go, Young Sam;Bae, Hwan Hee;Choi, Yu Chan;Son, Jae Han;Ha, Jun Young;Shin, Seong Hyu;Jung, Tae Wook
KOREAN JOURNAL OF CROP SCIENCE
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v.66
no.3
/
pp.240-247
/
2021
Anthocyanins are known to be involved in various functions such as antioxidant and antibacterial activities in plants. Although studies on anthocyanins in corn have been conducted recently, basic research related to anthocyanin biosynthesis is insufficient. In this study, we examined the molecular biological and physicochemical properties related to anthocyanin biosynthesis in the tassel and silks of Gwangpyeongok and Dacheongok cultivars. Anthocyanins were not synthesized in either the tassel or silks in Gwangpyeongok, whereas were synthesized in both in Dacheongok. The total anthocyanin content was approximately 30 times higher in the tassel and silks of Dacheongok than in those of Gwangpyeongok. In addition, C-3-G was measured only in the tassel of Dacheongok, and C-3-G, Pg-3-G, and M-3-G were 45.2 times, 27.3 times, and 37.6 times higher, respectively, in the silks of Dacheongok than of Gwangpyeongok. Expression of F3'H, DFR, and GST genes decreased in the tassel, and that of F3'H and DFR genes decreased in the silks of Gwangpyeongok. It was further confirmed that transcription factor P1 and R1 regulate the expression of anthocyanin biosynthetic genes in the tassel and silks, respectively, in Gwangpyeongok. Linoleic acid (C18:2) decreased by 6.6% and 10.9%, and linolenic acid (C18:3) increased by 8.5% and 8.5%, in the tassel and silks, respectively, of Gwangpyeongok compared to those of Dacheongok. Palmitic acid (C16:0) increased by 4.1% and oleic acid (C18:1) decreased by 2.1% in the silks of Gwangpyeongok compared to that in Dacheongok. In addition, the total fatty acid content in the tassel and silks increased by 10.3% and 30.4%, respectively, in Gwangpyeongok compared to that in Dacheongok. However, no significant results were observed in the analysis of phytosterol components. These results may be utilized as useful resources for the development of functional corn containing a large amount of anthocyanins.
In this study, the variation of 20 Pinus thunbergii isolates of F. circinatum from 2 damaged sites in Jeju-Island were compared with a known Fusarium circinatum using molecular biological techniques. Two- and four-year-old seedlings of Pinus rigida${\times}$Pinus x rigitaeda and two-, three- and six-year-old seedlings of P. thunbergii were inoculated with one of the most virulent isolates, FT-7, to determine differences in susceptibility. In site 1 (FT), 13 isolates of F. circinatum were isolated from 14 individuals and in site 2 (FS), 7 isolates of F. circinatum were isolated from 9 individuals. No difference was found in the sequences of the internal transcribed spacer (ITS) region of the ribosomal RNA genes in the FS and FT isolates, and also even in the known isolate of F. circinatum, FE 1-1. However, the ITS sequences of the FS and FT isolates differed from those of a fungus, Botrytis cinerea. Two-year-old seedlings of P. rigida${\times}$P. x rigitaeda showed higher susceptibility (93.3% of mortality) than four-year-old ones. Three-year-old seedlings of P. thunbergii showed the highest susceptibility (66.7% of mortality) compared to those at other ages in the same species. We found a positive correlation between basal diameter and lesion length in the seedlings of P. rigida${\times}$P. x rigitaeda ($R^2=0.66$) and P. thunbergii (p < 0.0001), respectively. There were significant differences in susceptibility by the age of seedlings in each of P. rigida${\times}$P. x rigitaeda (p < 0.0001) and P. thunbergii (p < 0.0001) based on lesion length.
Cognitive decline is characterized by reduced long-/short-term memory and attention span, and increased depression and anxiety. Such decline is associated with various degenerative brain disorders, especially Alzheimer's disease (AD) and Parkinson's disease (PD). The increases in elderly populations suffering from cognitive decline create social problems and impose economic burdens, and also pose safety threats; all of these problems have been extensively researched over the past several decades. Possible causes of cognitive decline include metabolic and hormone imbalance, infection, medication abuse, and neuronal changes associated with aging. However, no treatment for cognitive decline is available. In neurodegenerative diseases, changes in the gut microbiota and gut metabolites can alter molecular expression and neurobehavioral symptoms. Changes in the gut microbiota affect memory loss in AD via the downregulation of NMDA receptor expression and increased glutamate levels. Furthermore, the use of probiotics resulted in neurological improvement in an AD model. PD and gut microbiota dysbiosis are linked directly. This interrelationship affected the development of constipation, a secondary symptom in PD. In a PD model, the administration of probiotics prevented neuron death by increasing butyrate levels. Dysfunction of the blood-brain barrier (BBB) has been identified in AD and PD. Increased BBB permeability is also associated with gut microbiota dysbiosis, which led to the destruction of microtubules via systemic inflammation. Notably, metabolites of the gut microbiota may trigger either the development or attenuation of neurodegenerative disease. Here, we discuss the correlation between cognitive decline and the gut microbiota.
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