• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.49-56
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• 1993

현재 유구낭미충증을 혈청학적으로 진단하는 데에는 낭액항원을 이용한 특이항체검사법으로 micro-ELISA를 널리 이용하고 있다. 이 실험은 효소부착 이차항체를 사용하는 micro-ELISA방법 대신 민감도가 뛰어난 것으로 알려진 ABC-ELISA나 Protein-ELISA로 바꾸어 사용하면 검사의 민감도를 보다 개선할 수 있는지를 알기 위하여 실시하였다. ABC-ELISA에 의한 항체검사는 낭액항원 단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청 희석 1:10,000, biotinylated no-hmn IgG 1:100,000 회석, peroxidase conjugated streptavidin 1:40,000 희석 및 2,2-azino-di(3-ethylbenztlllazoline) sulfnnlc acid발색제를 사용하는 조건으로 실시하였고 415 nm에서 흡광도를 측정하였다. Protein A-ELISA는 항원단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청은 1:200 회석, HRP-Protein A 1:20,000 희석 및 ABTS 발색제를 사용하는 조건으로 실시하고 415 nm에서 흡광도를 측정하였다. 유구낭미충증 환자 115 명의 혈청을 검사한 바 민감도는 micro-ELISA 81.7%, Protein A-ELISA 82.6%, ABC-ELISA 86. 1%이었다. 다른 기생충성 질환자, 비기생충성 질환자 및 대조군 등 165명 혈청에서 특이도는 각각 88.5%, 93.3%, 93.8%이었다 세 가지 ELISA방법에 의한 항체가 사이에는 상관계수 0.84~0.86의 높은 상관관계가 성립하였다. 이상의 결과 ABC-ELISA는 micro-ELISA에 비하여 민감한 혈청학적 방법이고 실제 유구낭미충 특이항체 검사상의 특이도에서도 차이가 있다는 것을 알 수 있었다. 따라서 ABC-ELISA는 유구낭미충증의 혈청학적 진단에서 혈청 및 뇌척수액 등 검체를 적게 사용할 수 있다는 장점이 있다는 것을 알 수 있었다.

• 한국식품영양과학회지
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• 제23권6호
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• pp.945-949
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• 1994

Chondroitin sulfate (Chs) contents in edible snail , Achatina fulica Bowdich , andits processed meat extracts were determined by high-performance liquid chormatogrpahy(HPLC) and spectrophotometric method. Spectrophotometric method was based on the precipitation of acriflavine by ChS, and HPLC method was based on the detection of two unsaturated disaccharides, 2-acetamido-2-deoxy-3-O-($\beta$ -D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose($\Delta$Di-4S) and 2-acetamido-2deoxy-3-O-($\beta$-D-gluco-4-eepyranosyluronic acid)-6-O-sulfo-D-galactose ($\Delta$야-6S) librated from ChS byenzymeatic digestion with chondroitinase ABc. the ratio of 125$\mu$mol of sodium hydroxide to mg of ChS and 8$0^{\circ}C$ of reaction temperature were proper for alkaline hydrolysis to remove protein residue form ChS. In assay preparation for HPLC ethod, the iptimum concentration of the enzyme chondroitinase ABc was 0.15 unit per 50 $\mu\textrm{g}$ of ChS at a fixed reaction time (30 min) and pH 8.0 using Tris buffer. ChS content in edible snail was 177.6mg% by spectrophotometric method and 153.5mg% by HPLC method and those in the processed meat extract was 71.3mg% by spectrometric method ad 62.8mg% by HPLC method, respectively.

• 생명과학회지
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• 제34권2호
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• pp.86-93
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• 2024

플라스미드의 복제는 엄격하게 조절되어야 하기 때문에 일반적으로 rolling circle 복제를 수행하는 플라스미드들은 복제 개시인자인 RepB는 전사 및 번역 수준에서 엄격하게 조절되어 일정한 copy number를 유지한다. 플라스미드 pJB01에는 단일 오페론으로 구성된 세 개의 orfs (copA, repB, repC 또는 repABC)가 포함되어 있다. 아미노산 서열 분석에서 pJB01 CopA는 다른 플라스미드의 복제 수 조절 단백질로서 Cops와 상동성을 보였다. pMV158의 CopG와 비교할 때, CopA는 플라스미드의 일반화된 억제자의 모티브로 알려진 RHH (ribbon-helix-helix)를 형성하는 것으로 추정된다. gel mobility shift assay 결과 정제된 융합 단백질이 repABC 오페론의 operator 영역에 결합하는 것으로 나타났다. 전사 수준에 대한 CopA의 기능적 역할을 조사하기 위해 CopA R16M, K26R 및 E50V와 같은 세 개의 포인트 돌연변이가 CopA의 코딩 프레임에서 구성되었다. CopA R16M, K26R 및 E50V 돌연변이의 repABC mRNA 수준은 CopA wt보다 각각 1.84, 1.78 및 2.86배 증가했다. 또한 세 개의 CopA 유전자의 돌연변이로 인한 복제 수도 CopA wt보다 각각 1.86, 1.68 및 2.89배 증가했다. 이러한 결과는 CopA가 전사 억제자이며 복제 개시자로서 repABC mRNA 및 RepB 단백질 수를 감소시킴으로써 pJB01의 복제 수를 감소시키는 것으로 제의된다.

• Journal of Microbiology and Biotechnology
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• 제30권7호
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• pp.982-995
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• 2020

A putative multidrug efflux gene, yddA, was cloned from the Escherichia coli K-12 strain. A drug-sensitive strain of E. coli missing the main multidrug efflux pump AcrB was constructed as a host and the yddA gene was knocked out in wild-type (WT) and drug-sensitive E. coliΔacrB to study the yddA function. Sensitivity to different substrates of WT E.coli, E. coliΔyddA, E. coliΔacrB and E. coliΔacrBΔyddA strains was compared with minimal inhibitory concentration (MIC) assays and fluorescence tests. MIC assay and fluorescence test results showed that YddA protein was a multidrug efflux pump that exported multiple substrates. Three inhibitors, ortho-vanadate, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and reserpine, were used in fluorescence tests. Ortho-vanadate and reserpine significantly inhibited the efflux and increased accumulation of ethidium bromide and norfloxacin, while CCCP had no significant effect on YddA-regulated efflux. The results indicated that YddA relies on energy released from ATP hydrolysis to transfer the substrates and YddA is an ABC-type multidrug exporter. Functional study of unknown ATP-binding cassette (ABC) superfamily transporters in the model organism E. coli is conducive to discovering new multidrug resistance-reversal targets and providing references for studying other ABC proteins of unknown function.

• Natural Product Sciences
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• 제20권1호
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• pp.1-6
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• 2014

P-glycoprotein (P-gp) is a permeability glycoprotein also known as multidrug resistance protein 1 (MDR1). P-gp is an ATP-binding cassette (ABC) transporter that pumps various types of drugs out of cells. These transporters reduce the intracellular concentrations of drugs and disturb drug absorption. The Caco-2 cell permeability assay system is an effective in vitro system that predicts the intestinal absorption of drugs and the functions of enzymes and transporters. Rhodamine-123 (R-123) and digoxin are well-known P-gp substrates that have been used to determine the function of P-gp. Efflux of P-gp substrates by P-gp has been routinely evaluated. To date, a number of herbal medicines have been tested with Caco-2 cell permeability assay system to assess bioavailability. There are growing efforts to find phytochemicals that potentially regulate P-gp function. The Caco-2 cell permeability assay system is a primary strategy to search for candidates of P-gp inhibitors. In this mini review, we have summarized the P-gp modulation by herbal extracts, decoctions or single components from natural products using Caco-2 cell permeability assays. Many natural products are known to regulate P-gp and herbal medicines could be used in combination with conventional drugs to enhance bioavailability.

• 대한수의학회지
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• 제33권3호
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• pp.407-417
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• 1993

Bupleurum falcatum has been used for treatment of inflammation, jaundice, influenza and hepatitis as a traditional orient folk medicine. This experiment was carried out to evaluate the effect of B falcatum extract on cellular immune responses in vivo and in vitro. Antigen binding cell(ABC) assay, antibody production, Arthus and delayed-type hypersensitivity(DTH) reaction against sheep erythrocytes(SRBC) were very depressed in B falcatum extract treated group in vivo. The growth of Staphylococcus aureus in brain heart infusion(BHI) broth containing B falcatum extract was remarkably inhibited. Otherwise, that of Salmonella typhyimurium was not significantly increased in vitro. When B falcatum extract pretreated mice were intraperitoneally(IP) injected S typhimurium and S aureus, respectively, the number of bacteria in peritoneal exudates were time dependent declination compared with those of control, and the weight of spleen and the number of macrophage migration into peritoneal cavity have no difference from those of untreated control. B falcatum extract gradually increased phagocytic activities of peritoneal macrophage against Candida albicans time and dose dependently, and was not significant production of migration inhibiotory factor(MIF). But migration abilities of normal leucocytes in B falcatum extract pretreated group were decreased dose dependently. When B falcatum extract was IP administered, these data indicate that B falcatum extract increases level of serum coticosterone. Therefore, B falcatum extract was indirectly mediated in immune system by serum coticosterone having relation to immunosuppression. These results lead to the conclusion that B falcatum extract acts as a trigger or regulator of cellular immune responses in immune system.

• Archives of Pharmacal Research
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• 제23권2호
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• pp.182-186
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• 2000

Chondroitin sulfates proteoglycans were isolated from human placenta. For the identification of enzymatic digestion products of isolated proteoglycan, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. By the action of chondroitin ABC and chondroitin B lyase, three unsaturated disaccharides 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-D-galactose ($\delta$Di-OS), 2-acetamide-2-deoxy-3-O-($\beta$-D-gluco-4-enepyranosyluronic acid)-6-O-su lfo-D-galactose ($\delta$Di-6S) and 2-acetamide-2-deoxy-3-O-($\beta$-D-gl uco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ($\delta$Di-4S) were produced from the human placenta proteoglycan. The anticoagulant activity of chondroitin sulfate proteoglycan was evaluated by activated partial thromboplastin time (aPTT) assay and thrombin time (TT) assay. The clotting times of aPTT and TT were increased from 72 to 144 sec and 19 to 27 sec, respectively. The Immune-modulating activity of chondroitin sulfate proteoglycan was examined by cell proliferation assay and these results suggest that it may play a role in suppression of the function of immune-related cells.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.555-558
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• 1998

chondroitin sulfates were isolated from the mud snail. For the quantitative analysis of enzymatic digestion products of isolated chondroitin sulfates, strong anion exchange-high performance liquid chromatography (SAX-HPLC) was performed. by the action of chondroitinase ABC, three unsaturated disaccharides$ 2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-D-galactose $$({\Delta}Di-OS), $2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-6-O-sulfo-D-galactose ({\Delta}Di-6S) and 2-acetamide-2-deoxy-3-O-({\beta}-D-gluco-4-enepyranosyluronic acid)-4-O-sulfo-D-galactose ({\Delta}Di-4S)$ were produced from the mud snail chondroitin sulfates. The analysis showed that relative proportion of ${\Delta}Di-OS/{\Delta}Di-6S/{\Delta}Di-4S$ was 58.7/3.1/38.2. The immunomodulating activity of chondroitin sulfate was examined by cell proliferation assay and these results suggest that it might be a immunosuppressant.

• Journal of Microbiology and Biotechnology
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• 제31권4호
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• pp.520-528
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• 2021

Plesiomonas shigelloides, a member of the family Vibrionaceae, is a gram-negative, rod-shaped, facultative anaerobic bacterium with flagella. P. shigelloides has been isolated from such sources as freshwater, surface water, and many wild and domestic animals. P. shigelloides contains 102 O-antigens and 51 H-antigens. The diversity of O-antigen gene clusters is relatively poorly understood. In addition to O1 and O17 reported by other laboratories, and the 12 O serogroups (O2, O10, O12, O23, O25, O26, O32, O33, O34, O66, O75, and O76) reported previously by us, in the present study, nine new P. shigelloides serogroups (O8, O17, O18, O37, O38, O39, O44, O45, and O61) were sequenced and annotated. The genes for the O-antigens of these nine groups are clustered together in the chromosome between rep and aqpZ. Only O38 possesses the wzm and wzt genes for the synthesis and translocation of O-antigens via the ATP-binding cassette (ABC) transporter pathway; the other eight use the Wzx/Wzy pathway. Phylogenetic analysis using wzx and wzy showed that both genes are diversified. Among the nine new P. shigelloides serogroups, eight use wzx/wzy genes as targets. In addition, we developed an O-antigen-specific PCR assay to detect these nine distinct serogroups with no cross reactions among them.

• Toxicological Research
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• 제6권1호
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• pp.21-28
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• 1990

This study was undertaken to evaluate the immune responses to sheep red blood cell (SRBC) and potential anti-tumor effect of Bacillus Calmette-Guerin (BCG) in the mice bearing rumor induced by DMBA. The frequencies of tumor appearances were 62% in DMBA-treated mice and 14% in DMBA and BCG-treated group, respectively. Cellular immune response such as delayed-type hypersensitivity (DTH) to SRBCs, natural killer (NK) cell activity and antigen-binding cell (ABC) assay were decreased apparently in the tumor bearing mice compared to the normal controls. Humoral immune responses such as hemagglutinin (HA) and hemolysin (HE) were noted to be reduced in the tumor bearing mice, but the spleen index increased in tumor bearing mice. All the immunological parameters in the DMBA and BCG-group appeared to be higher than those of only DMBA-treated group. These results indicated that DMBA-induced tumor suppressed host immune responses. Also, they imply the idea that BCG enhanced the immune responses of tumor-bearing host and antitumor effects.