With the indirect immunobead antisperm antibody test(IBT) a prospective study was conducted to evaluate the immune status of 38 men before and after vasovasostomy. The pregnancy and postoperative semen analysis were evaluated. The results were compared between pregnant (n=14) and non-pregnant(n=24) group. The postoperative sperm motility was inversely correlated with the titer of the preoperative and postoperative IgG(p<0.01). The preoperative and postoperative titer of IgG were significantly higher than the titer of IgA or IgM(p<0.05). The mean percentage of the positive IBT(20 per cent binding or more) of the pregnant group was significantly lower than non-pregnant group in the preoprative and postoperative IgG(p<0.05). Immunobead binding restricted to the head and tail of a sperm in IgG was predominant and significantly lower in the pregnant group (p<0.05). In conclusion, IgG especially immunobead binding to the head and tail can be used as a sensitive screening assay for antisperm antibodies after vasovasostomy.
Two intraocular tumors were identified in low and medium dosed groups of a carcinogenicity study using 200 males and 200 females Sprague-Dawley rats in Screening & Toxicology Research Center, Korea Research Institute of Chemical Technology. The tumors were grossly observed as white or yellow, unilateral nodules. They were approximately $1-2{\times}3-5mm$ in size. The tumors located in the region of iris and/or ciliary body invaded peripheral cornea. The microscopic features were usually composed of spindle cells arranged in parallel, forming gently curving bundles or whorls. The spindle cells had poorly defined cell boundaries, scant to moderate cytoplasm, fusiform nuclei and indistinct nucleoli. Mitotic figures were rare and areas of necrosis were present. The spindle cells had positive immunoreactivity for S-100 protein and vimentin but negative for desmin, collagen and HMB-45 antibody. In special histochemical studies, the spindle cells react with Gomori's stain for argyrophih fibers, Prussian blue stain for iron but negative with Masson-Fontana's stain for melanin granules. Ultrastructurally, cytoplasmic premelanosomes were not observed in the tumor cells due to the poor preservation of tumor masses. Based on the results, the tumors were diagnosed as amelanotic melanoma.
Three dogs (An 8 years-old intact female Poodle, a 7 years-old intact male Schunauzer, and an 8 yearsold Golden Retriever) were presented due to acute vomiting, dyspnea, and generalized weakness. Megaesophagus was confirmed through radiographic examination in all 3 dogs. Relative oesophageal diameter (ROD) was measured and results of ROD measurements showed the possibility of megaesophagus secondary to myasthenia gravis in three dogs. Thus we performed anticholinesterase test as screening test for myasthenia gravis. In all three dogs, esophageal diameter was reduced after neostigmine methylsulfate administration. For definite diagnosis of acquired myasthenia gravis, serum acetylcholine receptor antibody titer was measured, but definite diagnosis was confirmed only in one case. However, based on history, radiographic findings, anticholinesterase test, ROD measurement, other two cases were still suspected as megaesophagus secondary to myasthenia gravis. Treatment with pyridostigmine bromide was initiated in all dogs, and improvement of esophageal diameter was shown in all dogs. One dog was successfully managed for 15 months after initial treatment and, is still alive, but other two dogs were died shortly after initial treatment, because of severe aspiration pneumonia.
We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.
The highest antitumor activity was observed in water soluble AS fraction of the Ganoderma lucidum IY 009. AS fraction did not show any cytotoxicity on sarcoma 180 cell but stimulated antibody production, opsonization of macrophage in ICR mouse and superoxide ion production from isolated macrophage. AS fraction activated complement C3 in human serum, and their antitumor activity was inhibited by EDTA, a chelator of cation related complementary activation. AS fraction exerted om prolong of life span and ingibition of tumor growth in the leukemia P388 or L1210 transplanted inbreed mouse,k BDF1 but krestin did not. AS fraction did not show any serious and lethal effects through oral administration on ICR mouse, and LD$_{50}$ of those was above 2,230 mg/kg.
A mycotoxin is a toxic secondary metabolite produced by organisms of the fungus kingdom, commonly known as molds and has been widely contaminated in agricultural products such as grains and cereals. Many methods including high performance liquid chromatography (HPLC) and gas chromatography (GC) have already been proposed and reviewed for mycotoxins. These methods are either expensive or time-consuming due to the complication of sample preparation and pre-concentration before determination. In addition, both methods are unsuitable for the routine screening of large sample numbers. A biosensor is a fictive analytical device that combines a biological component with a physicochemical detector for the detection of an analyte. Biosensors represent a rapidly expanding field, at the present time, with an estimated 60% annual growth rate; the major impetus coming from the health-care industry but with some pressure from other areas, such as food safety and environmental monitoring. Antibodies and aptamers are bioreceptors which have been used in the development of biosensors. There are many kinds of antibodies and aptamers specific to mycotoxin, and antibody (or aptamer)-based biosensors have been successfully developed for the detection of mycotoxin. The biosensors permit the rapid, sensitive, simple, and on-site detection of a range of mycotoxins and can be an alternative method to traditional methods such as HPLC and GC. This presentation provides the development trends of biosensors to mycotoxins and their application to food and agricultural products.
The Journal of the Korean Society for Microbiology
/
v.3
no.1
/
pp.15-23
/
1968
For the effective control of Syphilis, many investigators have developed a more rapid, simple and economical screening serological test which is adequately sensitive and specific. To fulfill the requirements of a more rapid serologic test for syphlis, a substitute for the conventional serum specimen was needed since considerable time and labor are involved in the processing of blood to serum. Burdon suggested the use of plasma in the serologic tests for syphilis as a substitute for serum. He noticed that plasma was more sensitive than serum in the Kline and Kahn tests, and attributed this to the presence of more antibody-like substance, "reagin" in plasma than in serum. However, to make plasma sufficiently sensitive, it was necessary to inactivate plasma by heating at a temperature of $56^{\circ}C$ for about 30 minutes. Heating of plasma resulted in the precipitation of fibrinogen which made centrifugation necessary to obtain dear plasma. Since the chief disadvantage to the use of unheated plasma(or serum) was a reduction in sensitivity of results-which probably was due to a labile factor such as complement-Portnoy et al began to consider rapid chemical methods of inactivation of plasma(or serum). They experienced that choline chloirde was shown to be anticomplementary which suggested its use as an inactivating agent for unheated plasma(or serum). In 1959 Portnoy et al reported the Rapid Plasma Reagin(RPR) Test for syphilis which is a more rapid, economical and simple. But still this test has many disadvantages as a rapid performing, field and office procedure, because it requires the usual laboratory equipments such as centrifuge, rotating machine, microscope etc. To substitute these disadvantages of the RPR test, in 1962, Portnoy et al developed the Rapid Plasma Reagin(RPR) card test for syphilis, which has the following advantages: a) Simplicity and rapidity of performance, b) Requires no laboratory equipments, c) Stable antigen suspension, d) Adequate sensitivity and specificity. This RPR card test can be used as a rapidly performing and screening test in field investigation, outpatient clinics, small laboratories and hospitals doing limited syphilis serology, and predonor in blood bank. Private clinic which has limited laboratory equipment and technic for syphilis serology can also use this RPR card test as a tool in the rapid diagnosis of syphilis. It was thought that this RPR card test is a useful tool in Korea for private physician and mass survey for syphilis diagnosis. But Portnoy patented the reagents needed for the performing the RPR card test. Therefore authors developed newly the reagents and according to Portnoy's method evaluated the newly developed. RPR card test compared with the VDRL, Kolmer CF, and RPCF tests. The RPR card and VDRL tests were performed plasma and serum from the total 1,132 cases. Among these 1,131 cases, 521 were syphilis suspected laboratory specimens, and 611 were syphilis unsuspected healthy young men. After screening with these two tests, the RPR card and VDRL tests, reactive specimens to the above one or both tests were retested by the Kolmer CF and RPCF tests.
Background: Hepatitis C virus(HCV), a family of Flaviviridae, has a host cell-derived envelope containing a positive-stranded RNA genome, and has been known as the maj or etiological agent for chronic hepatitis, hepatic cirrhosis, and hepatocellular carcinoma. There remains a need to dissect a molecular mechanism of pathogenesis for the development of therapeutic and effective preventive measure for HCV. Identification of cellular receptor is of central importance not only to understand the viral pathogenesis, but also to exploit strategies for prevention of HCV. This study was aimed at identifying peptide mimotopes inhibiting the binding of E2 protein of HCV to MOLT-4 cell. Methods: In this study, phage peptide library displaying a random peptides consisting of 7 or 12 random peptides was employed in order to pan against E2 protein. Free HCV particles were separated from the immune complex forms by immunoprecipitation using anti-human IgG antibody, and used for HCV-capture ELISA. To identify the peptides inhibiting E2-binding to MOLT-4 cells, E2 protein was subj ect to bind to MOLT-4 cells under the competition with phage peptides. Results: Several phage peptides were selected for their specific binding to E2 protein, which showed the conserved sequence of SHFWRAP from 3 different peptide sequences. They were also able to recognize the HCV particles in the sera of HCV patients captured by monoclonal antibody against E2 protein. Two of them, showing peptide sequence of HLGPWMSHWFQR and WAPPLERSSLFY respectively, were revealed to inhibit the binding of E2 protein to MOLT-4 cell efficiently in dose dependent mode. However, few membrane-associated receptor candidates were seen using Fasta3 programe for homology search with these peptides. Conclusion: Phage peptides containing HLGPWMSHWFQR and WAPPLERSSLFY respectively, showed the inhibition of E2-binding to MOLT-4 cells. However, they did not reveal any homologues to cellular receptors from GenBank database. In further study, cellular receptor could be identified through the screening of cDNA library from MOLT-4 or hepatocytes using antibodies against these peptide mimotopes.
Sera of cancer patients may contain antibodies that react with a unique group of autologous cellular antigens called tumor-associated antigens (TAAs). The present study aimed to determine whether a mini-array of multiple TAAs would enhance antibody detection and be a useful approach in esophageal cancer detection and diagnosis. Our mini-array of multiple TAAs consisted of eleven antigens, p53, pl6, Impl, CyclinB1, C-myc, RalA, p62, Survivin, Koc, CyclinD1 and CyclinE full-length recombinant proteins. Enzyme-linked immunosorbent assays (ELISA) were used to detect autoantibodies against eleven selected TAAs in 174 sera from patients with esophageal cancer, as well as 242 sera from normal individuals. In addition, positive results of ELISA were confirmed by Western blotting. In a parallel screening trial, with the successive addition of antigen to a final total of eleven TAAs, there was a stepwise increase in positive antibody reactions. The eleven TAAs were the best parallel combination, and the sensitivity and specificity in diagnosing esophageal cancer was 75.3% and 81.0%, respectively. The positive and negative predictive values were 74.0% and 82.0%, respectively, indicating that the parallel assay of eleven TAAs raised the diagnostic precision significantly. In addition, the levels of antibodies to seven antigens, comprising p53, Impl, C-myc, RalA, p62, Survivin, and CyclinD1, were significantly different in various stages of esophageal cancer, which showed that autoantibodies may be involved in the pathogenesis and progression of esophageal cancer. All in all, this study further supports our previous hypothesis that a combination of antibodies might acquire higher sensitivity for the diagnosis of certain types of cancer. A customized mini-array of multiple carefully-selected TAAs is able to enhance autoantibody detection in the immunodiagnosis of esophageal cancer and autoantibodies to TAAs might be reference indicators of clinical stage.
Heo, Seok-Mo;Lee, Sol;Wang, HongTao;Jeong, Jeong Hyeok;Oh, Sang Wook
Journal of Periodontal and Implant Science
/
v.46
no.5
/
pp.320-328
/
2016
Purpose: Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods: This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student's t-test. Results: Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434.10,139 ng/mL) and 8,598 ng/mL (range, 7,421.9,877 ng/mL), respectively. The Student's t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions: The presence of a significant difference in CSP1 levels between healthy subjects and periodontal patients suggests that CSP1 may be a potential biomarker for the detection or screening of periodontitis patients.
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