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

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

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: Prick test is most frequently used in clinical procedures because its simplicity and safety of application and better correlation with clinical history as well as specificity and high reproducibility. The chemiluminescent assay(CLA) is a new in vitro non-radioactive modification of radioallergosorbent test(RAST). The CLA permits quick, simultaneous detection of total and specific IgE in human serum. The CLA has shown good sensitivity, specificity, and a good correlation with the RAST and skin prick test. OBJECTIVE: The purpose of this study was to find causative allergens and to evaluate the clinical significance of CLA comparing the results of the CLA with those of the prick test in dermatologic outpatients. METHODS: We performed the prick test with 51 allergens(Bencard Ltd. Brentford, England) in 448 patients, and CLA with 35 allergens(Boehringer mannheim Korea. green cross inc.) in 280 patients with dermatologic diseases. RESULTS: 1. The positive reaction rate was highest in third decade and decreased over the age of 40 in prick and CLA. 2. The positive reaction rate of prick test(52.8%) was higher than that of CLA(25.0%). 3. The positive reaction rate to single allergens and compound allergens was 14.2% and 38.6% in the prick test and 2.5% and 22.5% in the CLA respectively. 4. In the prick test, the positive allergens, in descending order, were House dust 114(25.4%), D. farinae 88(19.6%), D. pteronyssinus 79(17.6%), Mixed inhalants 61(13.6%), and Threshing 47(10.5%). In the CLA Thistle 35(12.5%), Dandelion 31(11.1%), D. farinae 30(10.7%), D. pteronyssinus 29(10.4%), and Cat fur 12(4.3%). 5. The results of the CLA to common 2 allergens showed a concordance rate of 84.6%, sensitivity of 58.9%, and specificity of 86.1% compared with those of prick test. CONCLUSION: CLA may be a useful alternative method and shows a good correlation with the prick test. Combination of the two method results in a high diagnostic accuracy for the cause of allergic diseases.
Allergens ; Animals ; Cats ; Dust ; Humans ; Immunoglobulin E ; Korea ; Luminescent Measurements* ; Outpatients ; Sensitivity and Specificity ; Skin ; Taraxacum

Prostate-specific antigen(PSA) in serum exists in several molecular forms that can be measured by immunodetectable assays; free PSA, PSA complexed to alpha1-antichymotripsin and total PSA, which represents the sum of the free and complexed forms. We have determined the amount and ratio of these forms may be seful for increasing the ability of PSA to be used in distinguishing prostate cancer(PCa) from benign prostatic hyperplasia(BPH). We evaluated 117 asymptomatic healthy male controls(mean age 55.8, range 50-71) with no clinical evidence of PCa, 125 men with symptomatic BPH(mean age 67.6, range 52-89) who underwent prostatic surgery(open prostatectomy or TLRP) and 14 men with biopsy-proven, untreated PCa(mean age 71.3 range 54-86). Using ACS-PSA2 assay(Ciba-Corning). we measured the free PSA(F), total PSA(T) and free/total(F/T) ratio. The method utilized was monoclonal-polyclonal immunometric, chemiluminescent assay, using a standardized calibrator 90:10. We also performed total PSA measurement with Tandem-R PSA assay(Hybritech). Correlation between ACS-PSA2 assay and Tandem-RPSA assay was excellent (r2=0.93). Receivr operating characteristics(ROC) analyses between them showed very similar performances. ROC analysis of free to total PSA ratio was more superior than that of free form PSA only. When all subjects were included, the ratio of free to total PSA in controls, BPH and PCa were 0.27, 0.24 and 0.07, respectively. which significantly differentiated between PCa and benign conditions(p<0.01). When the cut-off value (Fr) was set into 0.12, its diagnostic sensitivity and specificity were 86% and 84%, respectively. These findings suggest that free to total PSA ratio may enhance the ability to distinguish benign histologic conditions from cancer.
Humans ; Luminescent Measurements ; Male ; Passive Cutaneous Anaphylaxis ; Prostate ; Prostate-Specific Antigen* ; Prostatectomy ; ROC Curve ; Sensitivity and Specificity

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: 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: 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