• Journal of Microbiology and Biotechnology
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• v.18 no.2
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• pp.263-269
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• 2008

Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.1
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• pp.42-46
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• 2009

Unlike most bacteria, Treponema pallidum subspecies cannot be readily isolated or sustained in cell culture for numerous generations. In korea, two non treponemal tests are currently considered as standard; the VDRL slide test and RPR card test. These tests are based on an antigen composed of an alcoholic solution containing measured amount of cardiolipin, cholesterol, and sufficient purified lecithin to produce reactivity. The nontreponemal reagin tests measure immunoglobulin M (IgM) and IgG to lipoidal material released from damaged host cells as well as to lipoprotein-like material and possibly by cardiolipin released from the treponemes. The object of the evaluation was to evaluate the performance of the Mediace RPR kit on the automated biochemistry analyzer system as a method for screen method of syphilis as well as to identify BFP possibility. For evaluation of routine screening test, a total 2,380 specimens tested by Mediace RPR from 28th Oct, 2007 to 22th Feb, 2008. For evaluation of BFP possiblility, we measured samples which have potential BFP reaction in Syphilis test such as ANA (anti-nuclear antibody) positive (135 samples), CRP (C-reactive protein) positive (100 samples), RF (Rheumatoid factor) positive (26 samples), and other potential BFP cases (17 samples) including total 278 samples. These samples were tested quantitative test Mediace RPR with Hitachi 7600 P module. For comparison with current manual test, VDRL slide test were performed. Of these 2380 specimens, 2350 were negative, 30 were positive, and one were positive with TPHA. Both methods agreed for 2356 (98.9%) samples. Of the 30 samples showed positive results over 1.0 R.U, 6 samples showed positive results with VDRL test. Of these 6 samples, 1 samples showed positive with TPHA test. The combination of the Automated Biochemistry analyzer and VDRL test for retest can be increase efficiency of syphilis screening test.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.2
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• pp.89-95
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• 2006

Potato plants carrying the Ry gene are extremely resistance to a number of potyviruses, but it is not known which variety expressed the resistance. In this investigation, combined classical and molecular techniques were used to identify virus resistance potatoes. Mechanical inoculation of 32 varieties of Korean potato cultivars, with potato virus Y (PVY), induced various symptoms, such as mosaic, yellowing, necrosis, mottle, vein clearing and vein bending. Different virus spreading patterns were observed, such as highly sensitive, moderate and resistant to $PVY^o$ inoculated leaves in different cultivars. From the results of double antibody sandwich-enzyme links immunosorbant assays (DAS-ELISA), coupled with reverse transcription polymerase chain reaction (RT-PCR), Winter valley and Golden valley were found to be highly susceptible and resistant cultivars to $PVY^o$ respectively. TEM was used as a complementary method to conform the localization of the virus in leaf tissues. TEM detect virus particles in Golden valley, where, ELISA and RT-PCR were unable to detect the CP gene. However, the interior part of the tissues was severely deformed in $PVY^o$ infected Winter valley, than Golden valley The Ry gene is involved in an induced response in $PVY^o$ infected Golden valley plants. The methods described in this study could be applied for the screening and development of antiviral potatoes.

• Korean Journal of Veterinary Service
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• v.35 no.2
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• pp.133-137
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• 2012

Brucellosis, a bacterial zoonoses caused by the genus Brucella is responsible for abortion and infertility in cow. Brucellosis is causing economic loss in dairy industries and prevalent worldwide including Bangladesh but limited studies are devoted to determine the prevalence and its association with reproductive factors of dairy cows in Bangladesh. Therefore, the present study was conducted to determine the seroprevalence of brucellosis in dairy cattle using screening test Rose Bengal test (RBT) and the positive sera were further confirmed by indirect- ELISA. For this purpose, a total of 400 serum samples from dairy cows with history of abortion and various reproductive disorders were collected from the Kurigram district of Bangladesh for the detection of Brucella antibody. The overall prevalence of brucellosis in dairy cattle was 2.25%. Brucellosis in cases of abortion and repeat breeding was 8.3% and 2.8%, respectively. The results shows higher prevalence of brucellosis in cases of abortion followed by repeat breeding, while there was no seropositive cases from other reproductive disorders. Age-wise sero-prevalence was found 3.0% in 2~3 years age group and 2.0% in 4~8 years age group. The prevalence of brucellosis in indigenous and cross-bred cattle was 3.6% and 1.7%, respectively. All the animals detected positive to brucellosis by RBT were not found to be positive by i-ELISA. However, the RBT might be a suitable screening test for the diagnosis of Brucella infection in field condition in Bangladesh. These data will help to develop effective disease prevention strategies.

• Animal cells and systems
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• v.13 no.3
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• pp.283-288
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• 2009

The ubiquitous plasma membrane $Na^+/H^+$ exchanger 1 (NHE1) is rapidly activated in response to various extracellular stimuli and maintains normal cytoplasmic pH. Yeast two-hybrid screening was used in order to identify proteins interacting with NHE1 using its cytoplasmic domain as a bait from HeLa cDNA library. One of the interacting cDNA clones was human Lactate dehydrogenase B (LDHB). In vitro translated LDHB was pulled down together with GST-NHE1.cd protein in the GST pull down assay, confirming the interaction in vitro. LDHB antibody immunoprecipitated endogenous LDHB together with NHE1 from H9c2 cells, validating cellular interaction between NHE1 and LDHB. Subsequent analysis revealed that the overexpression of LDHB increased intracellular PH, implying opening of the NHE1 transporter. Moreover, overexpression of LDHB activated caspase 3 and induced cell death, consistent with the expected phenotype of hyper-activation of NHE1. Collectively, our data indicate that LDHB modulates NHE1 activity via physical interaction.

• Environmental Analysis Health and Toxicology
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• v.19 no.4
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• pp.353-357
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• 2004

난황단백질인 비탈로제닌을 성숙한 암컷 붕어 혈청으로부터 음이온 교환 크로마토그래피를 이용하여 정제 하였다. 정제한 비탈로제닌을 BALB/c mice를 이용하여 폴리크로날 항체를 생산하였고 이를 protein A column을 사용하여 정제하였다. 또한 이렇게 정제된 폴리크로날 항체를 이용한 붕어 비탈로제닌 측정용 효소면역측정법을 개발하였으며 그 측정 범위는 2∼l,000ng/mL이고 recovery 변동 범위는 88∼112%였다. 또한 이 효소면역측정법을 평가하기 위해 성숙한 수컷 붕어를 1,000ng/L ethinylestradiol(EE$_2$) 에 4주 동안 노출시켜 유도되어지는 비텔로제닌을 측정하였다. 그 결과 성숙한 수컷 붕어의 경우 비탈로제닌이 3주 만에 암컷 붕어의 평균수치만큼 유도됨을 알 수 있었다.

• Bulletin of the Korean Chemical Society
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• v.18 no.1
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• pp.44-50
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• 1997

A method of dual-signal generation from two different enzymes was developed and utilized to simultaneously perform dual immunoassays in a single microwell. Two enzymes selected as tracers were horseradish peroxidase (HRP) and β-galactosidase (GAL). 3, 3', 5, 5'-Tetramethylbenzidine (TMB) and chlorophenolred-β-galactopyranoside (CPRG) as chromogenic substrates for the respective enzyme were used. Although the two enzymes showed their maximum activities at distinct pH conditions (pH 5.1 for HRP and 7.5 for GAL), the enzyme reactions were able to be concurrently carried out at pH 5.75 in a dual-substrate solution without signal loss. This performance was achieved by increasing TMB concentration two-fold, introducing potassium salt as activator of GAL reaction, and extending total reaction time 50%. The signal generation method was then used for dual-enzyme immunoassays to detect antibodies with co-immobilized Hepatitis C virus antigens (core and NS5) and a Hepatitis B virus antigen (PreS(2)) in a microwell. Dose-response curves of the assays revealed cooperativity between different antigen-antibody complex formation, which suggested that dual immunoassays can only be used for qualitative screening tests unless the antigens immobilized were spatially separated.

• Journal of Multimedia Information System
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• v.9 no.2
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• pp.161-170
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• 2022

Under General anesthesia with isoflurane, we insert a chicken's esophageal catheter into the near the left atrium. 1MHz radio wave was added to electrocardiogram electrodes of the esophagus, and the change of impedance (phase) was obtained by amplitude synchronous detection technique. At the same time, a thin tube is surgically inserted into the axillary artery to continuously measure blood pressure. The correlation between impedance (phase) and blood pressure was obtained. Both showed a very high correlation (R²=0.9665). It was also observed the waveform flowing from the left atrium into the left ventricle. When an individual infected with the avian influenza virus develops, the cytokine storms lead to hypotension earlier than the test for antigen-antibody reaction. In order to detect this, in the future, this impedance technique will be useful for screening individuals infected with avian influenza virus by measuring the blood pressure of chickens in cages in a non-contact manner using microwaves.

• Journal of Food Hygiene and Safety
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• v.3 no.2
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• pp.65-73
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• 1988

T-2 toxin is one of mycotoxins produced by fungi such as Fusarium spp. and possesses a potent cytotoxicity to eukaryotic cell. The contamination of mycotoxins in cereals and feedstuffs is one of the great concerns in health authorities. Therefore, the development of the specific, sensitive and simplified analysis method for T -2 toxin is required. During more than ten years, several chemical and biological analysis methods were proposed and applied for the detection and quantification of T-2 toxin. TLC, GLC-FID and GC-MS are widely employed, but these methods required numerous clean-up procedures before analysis, and the detection limit for T-2 toxin is more than 10 ppb. Biological analysis methods with dermal tissues and cultured cells are not specific to T-2 toxin, since T-2 toxin and other related derivatives possess a similar toxicological activity although their relative activity is different each otber. Based on tbe specific reaction between antibody and antigen, the authors tried to introduce the immunochemical methods for determination of T-2 toxin. The enzyme-linked immunosorbent assay method using monoclonal antibody for T-2 toxin was applied to analyse T-2 toxin. The detection limit of T-2 toxin by ELISA method was 0.1 ppb. The correlation between ELISA and GC-MS method on these samples was very high. ELISA method developed for the detection and quantification of T -2 toxin in this paper possesses simplicity, high sensitivity and specific for T-2 toxin. Furthermore, the ELISA method with T-2 toxin monoclonal antibody was an excellent tool for the screening of Fusarium spp. which was suspected to produce T-2 toxin.

• Laboratory Medicine Online
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• v.1 no.1
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• pp.64-66
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• 2011

Anti-Sd^a is of no clinical significance, because it rarely causes hemolytic transfusion reactions. Even when its presence is suspected during antibody screening test, further identification of the antibody is usually not performed. We experienced a case of anti-Sd^a in 73 yr-old male patient showing mixed field agglutination by microcolumn agglutination. Antibody specificity could not be identified by conventional antibody identification test, and it was proven to be anti-Sd^a by urine neutralization test. In spite of its little clinical significance, it may give incompatible crossmatching results reacting with Sd^a antigen, which occurs at a high frequency in general population. When incompatible crossmatch results arising from anti-Sd^a are suspected, the problem may be solved by using the urine-neutralized serum of in crossmatching test.