OBJECTIVEAn optimal automatic selection method of permissible source region is proposed to reduce the ill-conditioned and ill-posed problems in the reconstruction of the light source in bioluminescence tomography.

METHODSThe 2D images captured by CCD are mapped into surface light irradiance distribution based on the light propagating model. The relation matrix between the source and light distribution is obtained by finite element method. Permissive source region is determined by using the automatic selection method proposed in this paper, and then Tikhonov regularization is applied to reconstruct the light source.

RESULTSThe center point distance between the optimal permissible source region and true source is 1.26 mm, and the center point error of the reconstructed light source and true source is 0.47 mm, the volume error is 9.13 mm3.

CONCLUSIONThe optimal permissive source region selection strategy is effective to locate the permissive source region close to the true source, and reduces the reconstructed error due to subjective orientation of permissible source region. This proposed method is the basis of high precision source reconstruction in bioluminescence tomography.

Algorithms ; Light ; Luminescent Measurements ; Tomography

BACKGROUND: Multiple allergosorbent test chemiluminescent assay (MAST CLA) is a simple method for measuring total and allergen-specific IgE in human serum. Total IgE level, however, was much frequently high, even if no allergen-specific IgE could be detected in serum. The aim of this study was to evaluate the total IgE class of the MAST CLA system. METHODS: We studied 649 patients in whom MAST CLA (MAST Immunosystems, Inc., Mountain View, CA, USA) was tested and compared the results with those of total IgE, Phadiatop FEIA & RAST FEIA using Pharmacia CAP system (Pharmacia AB, Uppsala, Sweden). RESULTS: MAST CLA specific IgE was positive in 139 (21.4%) among 649 patients. Total IgE was increased over class 2 in 379 (74.3%) among 510 MAST CLA allergen-specific IgE -negative patients. Total IgE assayed by Pharmacia CAP system was increased over 100 kU/L in 33 (54.1%) among 61 MAST CLA allergen-specific IgE -negative patients. Especially, total IgE assayed by Pharmacia CAP system was increased over 100 kU/L in 25 (69.4%) of 36 patients when MAST CLA total IgE class is over 2. Phadiatop FEIA which is a screening test for inhalant allergy was positive in 11 (50.0%) of 22 MAST CLA allergen-specific IgE -negative patients, and especially, positive in 8 (66.7%) of 12 MAST CLA allergen-specific IgE -negative patients who had MAST CLA total IgE class over 2. Nine of 11 patients with positive Phadiatop FEIA were also found to be positive in d1 and/or d2 by RAST FEIA. CONCLUSIONS: It is concluded that another allergen screening tests should be used for detecting allergen-specific IgE, when MAST CLA total IgE is increased over class 2 with no detectable MAST CLA specific IgE because the sensitivity of MAST CLA allergen-specific IgE could be low.
Humans ; Hypersensitivity ; Immunoglobulin E* ; Luminescent Measurements ; Mass Screening

AIMTo develop a simple, fast and inexpensive approach as well as an instrument for detection of gene mutation.

METHODSPyrosequencing based on bioluminometry assay was employed to detect gene mutation. Pyrosequencing is a method of sequencing by synthesis step-by-step using four enzymes, DNA-polymerase, ATP sulfurylase, luciferase and apyrase. The signal was produced by detecting pyrophosphate released during a dNTP incorporation. For mutation detection, a DNA fragment was amplified by PCR at first, followed by a single-stranded DNA preparation. In the second step, a short primer was annealed to the position just before the mutation point. Finally, specific dNTPs were added in terms of the template sequence. The mutation species can be readily determined by the sequence.

RESULTSA new instrument was developed for gene mutation detection by pyrosequencing. To iteratively inject small amount of each dNTP for the sequencing reaction, capillaries were used to connect dNTP reservoirs and the reaction chamber. Each dNTP was delivered by adding a gas pressure on the top of a corresponding dNTP reservoir, by which 0.2 microL of dNTP can be exactly added each time. It was theoretically proved that undesired liquid seep through the capillary did not affect the sequencing reactions in pyrosequencing. In addition, the three possible variants (wildtype, mutant and heterozygote) of a mutant point Cys275Ser in P53 gene exon 8 were determined by pyrosequencing using the instrument. A simple method was also described for rapidly distinguishing the type of a variant.

CONCLUSIONThe developed method is very simple, and the corresponding instrument is inexpensive and easy to operate, which can be used to detect many types of mutation.

Exons ; genetics ; Genes, p53 ; genetics ; Humans ; Luminescent Measurements ; Point Mutation

BACKGROUND: Food allergies have been demonstrated to play an important role in the pathogenesis of atopic dermatitis(AD), affecting around 10 to 40% of patients with AD. While immediate-type clinical reactions to food can easily be identified, the evaluation of a food allergy in the absence of immediate clinical reactions still presents diagnostic difficulties. OBJECTIVE: The purpose of this study was to evaluate the diagnostic value of the atopy patch test with regard to late-phase reactions and to evaluate the relationship between patch test and specific serum IgE level. METHODS: We performed the patch test and chemiluminescent assay(CLA) for food antigens(cow's milk, soybean milk and hen's egg) in 45 patients with AD who were suspicious of food-related symptoms and 15 normal controls. RESULTS: 1. Positive reaction of CLA in 45 patients with AD were 7/18(38.9%) for the cow's milk, 5/14(35.7%) for the soybean milk and 4/13(30.7%) for the hen's egg. 2. Positive reaction of the patch test in patients with AD were 5/18(27.8%) for the cow's milk, 6/14(42.9%) for soybean milk and 4/13(30.7%) for hen's egg, but only one(6.7%) for cow's milk in 15 normal controls. 3. A highly significant correlation to food antigens existed between patch test and CLA, but one patient showed a positive reaction in the patch test for soybean milk despite the negative reaction in CLA. 4. There was no significant statistical correlation between the severity of AD and the positive reaction rate of the patch test. 5. There were no significant statistical differences between the age of AD patients and the positive reaction rate of the patch test. CONCLUSION: The atopy patch test seems to be a valuable tool in the diagnostic work-up of food allergies in patients with AD-especially with regard to late-phase clinical reactions.
Dermatitis, Atopic* ; Food Hypersensitivity ; Humans ; Immunoglobulin E* ; Luminescent Measurements ; Milk ; Ovum ; Patch Tests* ; Soybeans

BACKGROUND: The multiple allergosorbent test chemiluminescent assay (MAST-CLA) system is a simple method for measuring total and allergen-specific IgE antibodies in the serum of patients with atopic dermatitis. Total IgE, however, is much frequently higher, even if no allergen-specific IgE antibodies can be detected in serum. OBJECTIVE: The purpose of this study was to evaluate total IgE class, the allergen frequencies and their correlations in MAST-CLA in child and adult atopic dermatitis patients respectively. METHODS: A total of fifty two adult patients and ninety child patients with atopic dermatitis were evaluated by MAST-CLA between march 2002 and march 2005 at Soonchunhyang hospital. Positive rates of specific IgE and the total serum IgE level of the MAST-CLA allergy system were compared between child and adult patients. RESULTS: Among the subjects, 84.5% (80.0% for child patients and 92.3% for adult patients) of patients showed an elevated serum total IgE (more than class level 2) and 54.9% (45.6% for child patients, 71.2% for adult patients) revealed at least more than one allergen-specific IgE by MAST-CLA. The average 3.76 (3.39 for child patients, 4.16 for adult patients) different allergens was simultaneously detected in a single positive serum. Commonly-positive allergen rates, in descending order, were D. farinae 44.4%, D. pteronyssynos 38.7% and house dust 26.8%. Furthermore, the higher total serum IgE level in adult patients, the more probability of allergen-specific IgE positive results being disclosed (p<0.05). The number of allergen-specific IgE positive results was increased in a higher serum total IgE level (p<0.05). But total IgE positive rates which had allergen-specific antibody negative patients was 37.5% (45.8% for child patients, 25.0% for adult patients). Good correlation was obtained between total IgE levels and number of positive allergen-specific IgE in MAST-CLA, with 0.551 correlation coefficiency (p<0.05). CONCLUSION: Our study suggested that the MAST-CLA allergy system is a useful screening test to detect allergen- specific IgE and to evaluate patients with atopic dermatitis. But other allergen screening tests should be used for detecting allergens, when MAST-CLA total IgE class is increased over class 2 with no detectable specific IgE.
Adult* ; Allergens ; Antibodies ; Child* ; Dermatitis, Atopic* ; Dust ; Humans ; Hypersensitivity* ; Immunoglobulin E* ; Luminescent Measurements ; Mass Screening

BACKGROUND: The chemiluminescent assay(CLA) is a new in vitro non-radioactive modification of the radioallergosorbent test(RAST). The CLA permits quick, siultaneous detection of total and specific IgE in human serum up to 35 different allergens, as well as their semiquantitative concentrations into classes from 0(negative) to 4(very high). The CLA has shown sensitivity, specificity, and a good correlation with the RAST and also with skin pick testing. OBJECTIVE: The purpose of this study was to find the causatix llergens and to evaluate the clinical significance of CLA comparing the results of tbe CLA wi! h hose of the prick test in patients with chronic urticaria. METHODS: We performed the CLA with 35 allergens(MAST immunosunosystems, Inc., California, America) and the prick test with 51 allergens(Bencard Ltd., Bricantord, England) in 70 patients with chronic urticaria. The CLA detects the allergen-IgE reactic say the use of an enzyme-labeled anti-IgE and a coupled photoreagent mixture. lhe amount of the luminescent energy is measured by exposing a polarcid film and then semiquantitation is a sessed by using a densitometer. RESULTS: Except for those over the age of 60, all age-groups ealed a similar positive reaction to the CLA and prick test, but the positive reaction rate o the prick test(91.4%) was higher than that of the CLA(38.6%). The positive reaction rate oringle allergen and compound allergens was 10.0% and 28.6% in the CLA, but 10.0% and 81.4% in the prick test, respectively. In the CLA, the rate of positive reaction was 17(24.3%) in Dermatophagoides(D.) farinae, 15(21.4% ) in D. pteronyssinus, 5(7.1% ) in house dust, an 13(4.3%) in shrimp in that order. In the prick test, the rate of positive reaction was 39(55.7%) in house dust, 35(50.0%) in D. farinae, 29(41.4%) in D. pteronyssinus, and 21(30.0%) in haydrst(30.0%) in that order. The results of the CLA to 3 comrnon allergens showed a concordance rare of 67.6%, sensitivity of 34.3%, and specificity of 97.3% compared with those of the prick case. The levels of total IgE and eosinophils were elevated in 59(n=70, 843%) and in 5(n=56, 84.3%), respectively. CONCLUSION: The results of this study suggest that the CLA may be used as an alternative method for the diagnosis of chronic urticaria because it is and coiomic, safe, simple procedure with very high specificity and trelative low sensitivity, and is sirnilt. neously capable of testing to multiple allergens.
Allergens ; California ; Diagnosis ; Dust ; Eosinophils ; Humans ; Immunoglobulin E ; Luminescent Measurements* ; Sensitivity and Specificity ; Skin ; Urticaria*

BACKGROUND: Recently several papers have been published dealing with positive patch reactions to house dust mite antigens in atopic dermatitis patients. The possibility that house dust mite antigens, brought in direct contact with the skin such as through the air, may contribute in the pathogenesis of the dermatitis, is an interesting aspect of the relationship between atopic dermatitis and allergic reactions. OBJECTIVE: The purpose of this study is to evaluate type-IV and type-I hypersensitivity to house dust mite antigens in atopic dermatitis patients. METHODS: We performed the patch test and chemiluminescent assay(CLA) for house dust mite antigens in 42 patients with atopic dermatitis and 10 normal controls. RESULTS: 1. Positive reactions of the patch test in patients with atopic dermatitis were 11(26.2% ) for the Df antigen and 10(23.8%) for the Dp antigen. 2. Positive reactions of CLA in 42 patients with atopic dermatitis were 12(28.5%) for both Df and Dp antigen, but only one(10%) for both Df and Dp antigen in 10 normal controls. 3. A highly significant correlation to house dust mite antigens existed between the patch test and CLA, but some patients showed a positive reaction in the patch test despite the level of IgE which was in class 1 or 0. 4. There was no significant statistical correlation between the severity of atopic dermatitis and the positive reaction rate of both the patch test and CLA CONCLUSION: These results suggested that in atopic dermatitis patients with positive patch test reactions to house dust mite antigens, delayed type hypersensitivity reactions to these antigens might play an important role in the development of atopic dermatitis skin lesion .
Dermatitis ; Dermatitis, Atopic* ; Dust* ; Humans ; Hypersensitivity ; Immunoglobulin E* ; Luminescent Measurements ; Patch Tests* ; Pyroglyphidae* ; Skin

BACKGROUND: Multiple allergosorbent test chemiluminescent assay (MAST CLA) is a simple method for in vitro measurement of allergen-specific IgE antibodies. In the present study, the diagnostic performance of MAST CLA was evaluated from the data of Asan Medical Center. METHODS: With the test results of Korea IgE Panel, Korean Food Panel and Korean Inhalant Panel, we evaluated the next following. First, the change of the positive rates of allergen-specific IgE antibodies after the Korea IgE panel was divided into each Food Panel and Inhalant Panel; Second, the diagnostic performance of total IgE determined by MAST CLA in comparison with those determined by RIA; Third, the discrepancy of reactive intensity in Food and Inhalant Panels tested with the same specimens; Fourth, the diagnostic performance of MAST CLA compared with RAST, in detecting the IgE antibodies to three most common allergens. RESULTS: Overall positive rate was 33.9% (548/1,617); 39.3% for Korea IgE Panel, 22.9% for Food Panel, 34.6% for Inhalant Panel. Positive rate was increased only 0.5% for Food Panel and only 0.3% for Inhalant Panel by the new allergens added. The sensitivity, specificity, and concordance rate of total IgE levels determined by MAST CLA in comparison with those determined by RIA was 68.4%, 64.9%, 66.8% in Food Panel, and, 87.6%, 44.4%, 68.5% in Inhalant Panel. Five of the eighteen specimens tested by Food and Inhalant Panels simultaneously showed discrepant reactivity with the extent of class 1. The sensitivity, specificity, and concordance rate of MAST CLA compared with RAST were 36.1%, 100.0%, 67.1% for D. farinae, 30.8%, 96.8%, 60.6% for D. pteronyssinus, and 21.4%, 96.2%, 70.0% for housedust. CONCLUSIONS: The allergens newly added to Food and Inhalant Panels were not contributable to the detection of allergen-specific IgE. The specificity of total IgE was so low that positive total IgE result without specific IgE should be interpreted as false positive. The sensitivity to common allergens was also too low. So, MAST CLA needs further improvement to be used as a primary screening test for allergy.
Allergens ; Antibodies ; Chungcheongnam-do* ; Dust ; Hypersensitivity ; Immunoglobulin E ; Korea ; Luminescent Measurements* ; Mass Screening ; Sensitivity and Specificity

BACKGROUND: Urticaria and atopic dermatitis are representative allergic skin diseases that can be mediated by IgE. Measuring levels of specific IgE can be used to confirm causative agents of these skin diseases. OBJECTIVE: To analyze results from the multiple allergosorbent test chemiluminescent assay (MAST-CLA), which measures specific IgE in the presence of a causative agent/allergen, in IgE-mediated skin diseases. METHODS: A total of 404 patients with urticaria, atopic dermatitis or pruritus were enrolled in the present study. Positive rates of specific IgE as well as total serum IgE from the MAST-CLA were compared. RESULTS: Positive rates of specific IgE were highest in atopic dermatitis patients, followed by urticaria, and then pruritus, with 57.0%, 37.1%, and 20.8%, respectively (p<0.05). House dust mite species were the most common allergens in both atopic dermatitis and urticaria skin diseases. There were no differences in the overall MAST-CLA results between acute and chronic urticaria. The relative positive rate of inhalant allergen was significantly higher in adult than in child atopic dermatitis patients. CONCLUSION: Results from the MAST-CLA showed diversity among the three disease groups, and within each disease group, with different positive rates of specific IgE, a different mean allergen number per patient, and so on. Therefore, we concluded that MAST-CLA could be a useful diagnostic tool for various allergic skin diseases.
Adult ; Allergens ; Child ; Dermatitis, Atopic ; Humans ; Immunoglobulin E ; Luminescent Measurements ; Pruritus ; Pyroglyphidae ; Skin ; Skin Diseases ; Urticaria

AIMTo study the sensitizing effect of acemetacin (ACE) on the weak chemiluminescent (CL) reaction of KMnO4 with sulfite and establish a fast and convenient method for CL detection of ACE.

METHODSUsing the sensitizing effect of ACE on KMnO4-Na2SO3 system and flow injection technique to determine the concentration of ACE.

RESULTSUnder optimal conditions, the CL intensity of 1.0 x 10(-2) mol x L(-1) H3PO4 - 5.0 x 10(-5) mol x L(-1) KMnO4 - 4.0 x 10(-4) mol x L(-1) Na2SO3 was proportional to the concentration of ACE ranging from 1.0 x 10(-7) to 1.0 x 10(-5) mol x L(-1). The detection limit of ACE was 6.9 x 10(-8) mol x L(-1) at 3sigma. Satisfactory results were obtained for determination of ACE at 2.5 x 10(-6) mol x L(-1).

CONCLUSIONThe present method showed good precision, high sensitivity and selectivity and could be used for fast and convenient detection of ACE. It would be of significance to the clinical and pharmacological study of acemetacin.

Flow Injection Analysis ; methods ; Indomethacin ; analogs & derivatives ; analysis ; Luminescent Measurements ; methods ; Potassium Permanganate ; chemistry ; Sulfites ; chemistry