The cytotoxicity of medical ultrasonic couplant was tested by MTT assay and agar overlay test. By MTT assay, when the inoculum density was high, the cytotoxicity level was low, or vice versa. The cytotoxicity grade tested by agar overlay was not accord to MTT assay's too. MTT assay is suitable to test the cytotoxicity of medical ultrasonic couplant because it is quantitative and more sensitive, however, the experimental condition and the preparative method of extraction should be determined.
Animals ; Cell Line ; Colorimetry ; Cytotoxicity Tests, Immunologic ; methods ; Mice ; Ultrasonics

OBJECTIVETo establish a cell-based detection method of ciguatoxin using fluorescence assay.

METHODSMouse neuroblastoma N-2A cells were exposed to ouabain and veratridine and different concentrations of standard ciguatoxin samples (P-CTX-1) to establish the curvilinear relationship between the toxin dosage and fluorescence intensity using the sodium fluorescence probe CoroNaTM Green. The toxicity curvilinear relationship was also generated between the toxin dosage and cell survival using CCK-8 method. Based on these standard curves, the presence of ciguatoxin was detected in 33 samples of deep-sea coral fish.

RESULTSA correlation was found between the detection results of cell-based fluorescence assay and cytotoxicity assay, whose detection limit reached 103 g/ml and 1012 g/ml, respectively. The cell-based fluorescent assay sensitivity showed a higher sensitivity than cytotoxicity assay with a 2-4 h reduction of the detection time.

CONCLUSIONSThe cell-based fluorescent assay can quickly and sensitively detect ciguatoxin and may serve as a good option for preliminary screening of the toxin.

Animals ; Cell Line, Tumor ; Ciguatoxins ; toxicity ; Cytotoxicity Tests, Immunologic ; methods ; Fishes ; Fluorescent Dyes ; Mice ; Sodium

Hypersensitivity reaction to progesterone is a rare pathologic condition which consists of autoimmune response to endogenous progesterone, known as autoimmune progesterone dermatitis, and hypersensitivity reaction to exogenous progestogen. We report the case of a 31-year-old woman with a history of whole body urticaria during exogenous progesterone supplementation for in vitro fertilization (IVF). She was admitted to the hospital for the diagnosis and management of progestogen hypersensitivity. An intradermal test with progesterone revealed positivity to 5 mg/mL of progesterone. For her next IVF, progesterone desensitization was performed in a method combining oral and intramuscular progesterone administration. After successfully achieving a target dose of 100 mg per day, the route of progesterone administration was converted to intravaginal tablet (90 mg twice a day) without any hypersensitivity reactions.
Adult ; Autoimmunity ; Dermatitis ; Desensitization, Immunologic ; Diagnosis ; Drug Hypersensitivity ; Female ; Fertilization in Vitro ; Humans ; Hypersensitivity* ; Intradermal Tests ; Methods ; Progesterone* ; Urticaria

The natural killer(NK) cell activity of mononuclear cells (MNC) from peripheral blood (PB) and synovial fluid (SF) of 40 rheumatoid arthritis(RA) patients was investigated by employing 51-chromium-(51Cr) release microcytotoxicity and single cell cytotoxicity assays against K562 target cells. It has been revealed that SF-MNC from RA patients showed a significantly lower NK activity than PB-MNC from the same patients and this might be due to an impaired target binding capacity of the effector cells and not due to a deficiency of active NK cells.
Adolescent ; Adult ; Aged ; Arthritis, Rheumatoid/*immunology ; Chromium Radioisotopes/diagnostic use ; Comparative Study ; Cytotoxicity Tests, Immunologic/methods ; *Cytotoxicity, Immunologic ; Female ; Human ; In Vitro ; Killer Cells, Natural/*immunology ; Male ; Middle Age ; Support, Non-U.S. Gov't ; Synovial Fluid/immunology

BACKGROUND: In vitro serum allergen-specific IgE tests have been routinely used in the clinical diagnosis of allergic diseases. We evaluated the clinical usefulness of a newly introduced multiple antigen screen test, Advansure Allergy Screen (LG Life Science, Korea) (LG-Screen) for the detection of allergen specific IgE. METHODS: A total of 180 sera (80 for inhalant and 100 for food panels) were tested by LG-Screen and RIDA Allergy Screen (R-biopharm, Germany) (RIDA-Screen) assays. According to the 58-60 specific allergens or allergen groups, the positive rates and agreement rates were analyzed using the cut off levels of class 2. For the quantitation of total IgE and specific IgE, nephelometry and ImmunoCAP test were performed in the sera showing discrepant results between the two allergy screen assays. RESULTS: The agreement rate and kappa value (k) of total IgE between the two allergy screen assays was 73.9% and 0.333. LG-Screen showed higher agreement rate with nephelometry than RIDA-Screen. The positive rates to common outdoor inhalant and food allergens were significantly higher in RIDA-Screen. Overall agreement rate of specific IgE between the two allergy screen assays for 58 allergens was 86.7% (6,086/7,020) (k, 0.293). In samples showing discrepant results between the two allergy screen assays, concordance rate of allergy screen assay with ImmunoCAP assay was 70.9% (449/633) for LG-Screen (k, 0.585) and 29.1% (184/633) for RIDA-Screen (k, -0.303). CONCLUSIONS: LG-Screen showed a favorable agreement with RIDA-Screen and ImmunoCAP assays, and it could be used for the detection of allergen specific IgE in the clinical laboratory.
Adolescent ; Adult ; Aged ; Allergens/diagnostic use/*immunology ; Child ; Child, Preschool ; Female ; Humans ; Hypersensitivity, Immediate/diagnosis ; Immunoglobulin E/*blood ; Immunologic Tests/methods ; Infant ; Male ; Middle Aged ; Reagent Kits, Diagnostic

Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; immunology ; physiology ; Humans ; Immunologic Tests ; methods ; Virus Replication

aboratory diagnosis of tuberculosis (TB) traditionally relies on smear microscopy and culture of Mycobacterium tuberculosis from clinical samples. With recent advances in technology, there have been numerous efforts to develop new diagnostic tests for TB that overcome the low sensitivity and specificity and long turnover time associated with current diagnostic tests. Molecular biological tests based on nucleic acid amplification have brought an unprecedented opportunity for the rapid and specific detection of M. tuberculosis from clinical specimens. With automated sequencing analysis, species identification of mycobacteria is now easier and more accurate than with conventional methods, and rapid detection of mutations in the genes associated with resistance to TB drugs provides early information on the potential drug resistance for each clinical isolate or for clinical samples. In addition, immunological tests for the detection of M. tuberculosis antigens and antibodies to the antigens have been explored to identify individuals at risk of developing TB or with latent TB infection (LTBI). The recent introduction of commercial IFN-gamma assay kits for the detection of LTBI provides a new approach for TB control even in areas with a high incidence of TB. However, these molecular and immunological tools still require further evaluation using large scale cohort studies before implementation in TB control programs.
Antigens, Bacterial/immunology ; DNA, Bacterial/chemistry/genetics ; Humans ; Immunologic Tests/*methods ; Interferon-gamma/analysis ; Mycobacterium tuberculosis/genetics/immunology ; Sequence Analysis, DNA ; Tuberculin Test ; Tuberculosis/*diagnosis/immunology/microbiology

aboratory diagnosis of tuberculosis (TB) traditionally relies on smear microscopy and culture of Mycobacterium tuberculosis from clinical samples. With recent advances in technology, there have been numerous efforts to develop new diagnostic tests for TB that overcome the low sensitivity and specificity and long turnover time associated with current diagnostic tests. Molecular biological tests based on nucleic acid amplification have brought an unprecedented opportunity for the rapid and specific detection of M. tuberculosis from clinical specimens. With automated sequencing analysis, species identification of mycobacteria is now easier and more accurate than with conventional methods, and rapid detection of mutations in the genes associated with resistance to TB drugs provides early information on the potential drug resistance for each clinical isolate or for clinical samples. In addition, immunological tests for the detection of M. tuberculosis antigens and antibodies to the antigens have been explored to identify individuals at risk of developing TB or with latent TB infection (LTBI). The recent introduction of commercial IFN-gamma assay kits for the detection of LTBI provides a new approach for TB control even in areas with a high incidence of TB. However, these molecular and immunological tools still require further evaluation using large scale cohort studies before implementation in TB control programs.
Antigens, Bacterial/immunology ; DNA, Bacterial/chemistry/genetics ; Humans ; Immunologic Tests/*methods ; Interferon-gamma/analysis ; Mycobacterium tuberculosis/genetics/immunology ; Sequence Analysis, DNA ; Tuberculin Test ; Tuberculosis/*diagnosis/immunology/microbiology

PURPOSE: To evaluate the diagnostic utility and predictors for determinate results of an enzyme-linked immunospot assay using induced sputum cells (IS ELISPOT) for a rapid diagnosis of pulmonary tuberculosis (TB). MATERIALS AND METHODS: Subjects suspected of pulmonary TB who had either sputum acid fast bacilli smear-negative or not producing sputum spontaneously were prospectively enrolled. ELISPOT assay was performed using cells from induced sputum. RESULTS: A total of 43 subjects, including 25 with TB (TB group) and 18 with non-TB disease (non-TB group) were enrolled. Results of IS ELISPOT were determinate in only 17/43 (39%) subjects, but all of determinate results were consistent with the final diagnosis. Of the 43 sputum samples, 11 (26%) were inadequate to perform IS ELISPOT. Of 32 adequate sputum samples, the proportion of determinate results was significantly higher in the TB group (75%, 15/20) than in the non-TB group (17%, 2/12) (p=0.002). The status of active TB was a unique predictor but smear positivity was not a significant predictor for determinate results. In addition, sensitivity of IS ELISPOT (75%, 9/12) in smear negative TB was higher than that of TB-polymerase chain reaction (25%, 3/12). CONCLUSION: IS ELISPOT showed relatively high diagnostic value and accuracy in the TB group, independent of smear positivity. IS ELISPOT may provide additional diagnostic yield for microbiological tools in the rapid diagnosis of smear-negative TB.
Adult ; Aged ; *Enzyme-Linked Immunospot Assay ; Female ; Humans ; Immunologic Tests/*methods ; Male ; Middle Aged ; Mycobacterium tuberculosis/*isolation & purification ; Predictive Value of Tests ; Prospective Studies ; Reproducibility of Results ; Risk Factors ; Sensitivity and Specificity ; Sputum/*microbiology ; Tuberculosis, Pulmonary/*diagnosis/microbiology

TNF-related apoptosis-inducing ligand (TRAIL) is a member of factor TNF family, which could be potentially developed as novel antitumor agent due to its selective and efficient induction of apoptosis in tumor cells. Gene recombinant expression is an important tool for production of pharmaceutical protein. In this paper, the gene encoding human soluble TRAIL (114-281aa fragment) was cloned by PCR and then inserted into the Pichia Pastoris expression vector pPIC9K. The transformants were double-screened on plates containing neomycin G418 and many clones with high levels of G418-resistance were selected for further studies on protein expression. The recombinant human soluble TRAIL was secreted into the BMMY media under the condition of 3% methanol. And the recombinant protein was purified to homogeneity (-80% purity) by using Ni-agarose affinity chromatography. The yield of this protein is about 1-2 mg per liter culture. Cell viability assays demonstrated that human soluble TRAIL was cytotoxic in both leukemia cells Jurkat and lung cancer cells A549. After treatment with 0.05 microg/ml TRAIL, the survival rate of Jurkat cells was about 10%. The expressed TRAIL showed dose-dependent cytotoxicity in A549 cells within the range of 0.1-1 microg/ml. When the protein concentration reached 1 microg/ml, the survival rates of A549 cells were about 30%. However, the recombinant human soluble TRAIL did not show obvious cytotoxicity in human skin fibroblast cells (HSF) at concentrations tested. There results demonstrate that human soluble TRAIL is selectively cytotoxic in tumor cells. The expression system constructed in this experiment might contribute to further production of soluble TRAIL and TRAIL-based novel fusion proteins in large quantities.
Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cloning, Molecular ; Cytotoxicity Tests, Immunologic ; methods ; Genetic Vectors ; genetics ; Humans ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; biosynthesis ; genetics