A New Approach to Quantify and Grade Radiation Dermatitis Using Deep-Learning Segmentation in Skin Photographs.

AIMS: Objective evaluation of radiation dermatitis is important for analysing the correlation between the severity of radiation dermatitis and dose distribution in clinical practice and for reliable reporting in clinical trials. We developed a novel radiation dermatitis segmentation system based on convolutional neural networks (CNNs) to consistently evaluate radiation dermatitis. MATERIALS AND METHODS: The radiation dermatitis segmentation system is designed to segment the radiation dermatitis occurrence area using skin photographs and skin-dose distribution. A CNN architecture with a dilated convolution layer and skip connection was designed to estimate the radiation dermatitis area. Seventy-three skin photographs obtained from patients undergoing radiotherapy were collected for training and testing. The ground truth of radiation dermatitis segmentation is manually delineated from the skin photograph by an experienced radiation oncologist and medical physicist. We converted the skin photographs to RGB (red-green-blue) and CIELAB (lightness (L∗), red-green (a∗) and blue-yellow (b∗)) colour information and trained the network to segment faint and severe radiation dermatitis using three different input combinations: RGB, RGB + CIELAB (RGBLAB) and RGB + CIELAB + skin-dose distribution (RGBLAB_D). The proposed system was evaluated using the Dice similarity coefficient (DSC), sensitivity, specificity and normalised Matthews correlation coefficient (nMCC). A paired t-test was used to compare the results of different segmentation performances. RESULTS: Optimal data composition was observed in the network trained for radiation dermatitis segmentation using skin photographs and skin-dose distribution. The average DSC, sensitivity, specificity and nMCC values of RGBLAB_D were 0.62, 0.61, 0.91 and 0.77, respectively, in faint radiation dermatitis, and 0.69, 0.78, 0.96 and 0.83, respectively, in severe radiation dermatitis. CONCLUSION: Our study showed that CNN-based radiation dermatitis segmentation in skin photographs of patients undergoing radiotherapy can describe radiation dermatitis severity and pattern. Our study could aid in objectifying the radiation dermatitis grading and analysing the reliable correlation between dosimetric factors and the morphology of radiation dermatitis.

Human CD4+ CD39+ regulatory T cells produce adenosine upon co-expression of surface CD73 or contact with CD73+ exosomes or CD73+ cells.

While murine CD4(+) CD39(+) regulatory T cells (T(reg)) co-express CD73 and hydrolyze exogenous (e) adenosine triphosphate (ATP) to immunosuppressive adenosine (ADO), surface co-expression of CD73 on human circulating CD4(+) CD39(+) T(reg) is rare. Therefore, the ability of human T(reg) to produce and utilize ADO for suppression remains unclear. Using mass spectrometry, we measured nucleoside production by subsets of human CD4(+) CD39(+) and CD4(+) CD39(-)CD73(+) T cells or CD19(+) B cells isolated from blood of 30 volunteers and 14 cancer patients. CD39 and CD73 expression was evaluated by flow cytometry, Western blots, confocal microscopy or reverse transcription-polymerase chain reaction (RT-PCR). Circulating CD4(+) CD39(+) T(reg) which hydrolyzed eATP to 5'-AMP contained few intracytoplasmic granules and had low CD73 mRNA levels. Only ∼1% of these T(reg) were CD39(+) CD73(+) . In contrast, CD4(+) CD39(neg) CD73(+) T cells contained numerous CD73(+) granules in the cytoplasm and strongly expressed surface CD73. In vitro-generated T(reg) (Tr1) and most B cells were CD39(+) CD73(+) . All these CD73(+) T cell subsets and B cells hydrolyzed 5'-AMP to ADO. Exosomes isolated from plasma of normal control (NC) or cancer patients carried enzymatically active CD39 and CD73(+) and, when supplied with eATP, hydrolyzed it to ADO. Only CD4(+) CD39(+) T(reg) co-incubated with CD4(+) CD73(+) T cells, B cells or CD39(+) CD73(+) exosomes produced ADO. Thus, contact with membrane-tethered CD73 was sufficient for ADO production by CD4(+) CD39(+) T(reg). In microenvironments containing CD4(+) CD73(+) T cells, B cells or CD39(+) CD73(+) exosomes, CD73 is readily available to CD4(+) CD39(+) CD73(neg) T(reg) for the production of immunosuppressive ADO.

YKL-40 in induced sputum after allergen bronchial provocation in atopic asthma.

BACKGROUND: Serum chitinase-like proteins such as YKL-40 in asthmatic patients are known to positively correlate with disease severity but controversy remains regarding their role. The allergen bronchial provocation test (ABPT) can induce allergic airway inflammation in individuals with atopic asthma. OBJECTIVE: To evaluate the induction and kinetics of YKL-40 during allergen-induced airway inflammation in atopic asthmatics. METHODS: Thirteen patients were enrolled from May to November 2008. They all underwent ABPT with Dermatophagoides farinae crude extract. Induced sputums (IS) and serum were collected 3 times: 7 days before ABPT (baseline), 7 hours after ABPT, and 24 hours after ABPT. We examined the cytology of induced sputum (IS) and measured levels of YKL-40, interleukin (IL) 4, IL-5, IL-13, IL-33, tumor necrosis factor (TNF) alpha, and eosinophilic cationic protein (ECP) in IS and/or serum. RESULTS: Following ABPT, total inflammatory cells, eosinophils, and neutrophils increased in a time-dependent manner in IS. YKL-40 levels were increased in IS but not in serum at 7 or 24 hours after ABPT (P=.011 and P=.006, respectively). Similarly to YKL-40, IL-5 and ECP levels were also increased in IS at 7 and 24 hours after ABPT (P=.011 for IL-5 and P=.006 for ECP). Overall, YKL-40 levels were well correlated with ECP levels in IS (p=0.576, P<.001). CONCLUSIONS: YKL-40 levels increased immediately in IS but not in the serum of atopic asthmatics. The correlation between YKL-40 levels and ECP in IS suggests that YKL-40 may play a pathophysiologic role in human atopic asthma.

Ectopic matrix metalloproteinase-9 expression in human brain tumor cells enhances oncolytic HSV vector infection.

Oncolytic herpes simplex virus (oHSV) vectors have shown promise in the treatment of patients with recurrent brain tumors although few complete responses have accrued. Impediments to effective therapy include limited vector distribution on delivery, a consequence of injected virion particle trapping in the tumor extracellular matrix (ECM). To enhance virus delivery and spread, we investigated the use of the matrix metalloproteinase-9 (MMP-9) as a means to degrade collagen type IV, a major component of the ECM and basement membranes of gliomas that is absent in normal brain tissue. SK-N-AS neuroblastoma cells were transduced for constitutive, elevated expression of MMP-9, which did not enhance tumor cell migration in vitro or tumor progression in a murine xenograft brain tumor model. MMP-9 expression improved the distribution and infection of oHSV vectors in spheroid model in vitro. Furthermore, MMP9 induced a vector infection over larger areas of brain tumors in vivo. These results suggest that vector delivery and distribution in vivo can be improved by compromising the ECM, potentially enhancing oncolytic efficacy.

Relationship between neurokinin 2 receptor gene polymorphisms and serum vascular endothelial growth factor levels in patients with toluene diisocyanate-induced asthma.

BACKGROUND: Among the various pathogenic mechanisms of toluene diisocyanate (TDI)-induced asthma, a contribution from neurogenic inflammation has been suggested. OBJECTIVE: To evaluate neurokinin 2 receptor (NK2R) gene polymorphisms in association with the clinical phenotype of TDI-induced asthma, 70 TDI-induced occupational asthma (TDI-OA)patients, 59 asymptomatic exposed controls (AEC), and 93 unexposed healthy controls (NC) were enrolled in the study. METHODS: Two single-nucleotide polymorphisms (SNPs) of NK2R, 7853G>A (Gly231Glu) and 11 424G>A (Arg375His), were genotyped using a single base extension method. The levels of PC20 methacholine, specific IgE and IgG to TDI-human serum albumin conjugate, and serum vascular endothelial growth factor (VEGF), matrix metalloproteinase-9, and TGF-beta1 were compared according to the NK2R genotypes of the subjects with TDI-OA and AEC. RESULTS: No significant differences in allele, genotype, or haplotype frequencies of these two SNPs were noted among the three groups (P>0.05, respectively). Moreover, subjects with the NK2R 7853GG genotype had higher serum VEGF levels than those with GA or AA among the TDI-exposed workers (P=0.040). CONCLUSION: The NK2R 7853GG genotype may contribute to increased serum VEGF levels, which result in airway inflammation after TDI exposure.

Herpes simplex virus RNAi and neprilysin gene transfer vectors reduce accumulation of Alzheimer's disease-related amyloid-beta peptide in vivo.

Accumulation of insoluble aggregates of amyloid-beta peptide (Abeta), a cleavage product of amyloid precursor protein (APP), is thought to be central to the pathogenesis of Alzheimer's disease (AD). Consequently, downregulation of APP, or enhanced clearance of Abeta, represent possible therapeutic strategies for AD. We generated replication-defective herpes simplex virus (HSV) vectors that inhibit Abeta accumulation, both in vitro and in vivo. In cell culture, HSV vectors expressing either (i) short hairpin RNA directed to the APP transcript (HSV-APP/shRNA), or (ii) neprilysin, an endopeptidase that degrades Abeta (HSV-neprilysin), substantially inhibited accumulation of Abeta. To determine whether these vectors showed similar activity in vivo, we developed a novel mouse model, in which overexpression of a mutant form of APP in the hippocampus, using a lentiviral vector (LV-APP(Sw)), resulted in rapid Abeta accumulation. Co-inoculation of LV-APP(Sw) with each of the HSV vectors showed that either HSV-APP/shRNA or HSV-neprilysin inhibited Abeta accumulation in this model, whereas an HSV control vector did not. These studies demonstrate the utility of HSV vectors for reducing Abeta accumulation in the brain, thus providing useful tools to clarify the role of Abeta in AD that may facilitate the development of novel therapies for this important disease.

Allergen-specific immunosuppression by ovalbumin fused with diphtheria toxin in mice sensitized with albumins of different origin.

BACKGROUND: We previously reported that ovalbumin-diphtheria toxin (OVA-DT) fusion protein eliminates mast cells bearing OVA-specific IgE and protects OVA-sensitized mice from fatal anaphylaxis induced by OVA challenge. OBJECTIVE: To prove the specificity of therapeutic effect of OVA-DT to allergy induced by OVA only and not by other allergens such as human serum albumin (HSA), and to examine the cytotoxic effect of OVA-DT on B cells bearing OVA-specific IgE. METHODS: Mice were sensitized with two different antigens, OVA and HSA, and then treated with OVA-DT. The therapeutic effect of OVA-DT on the allergy response to each of allergen was evaluated by anaphylactic test. The effect of OVA-DT on the production of allergen-specific Ig isotypes of the sensitized mice and the cytotoxic effect of OVA-DT on B cells expressing OVA-specific IgE were examined. RESULTS: OVA-DT suppressed only OVA-induced allergy but not HSA-induced allergy in mice sensitized with a mixture of OVA and HSA. The suppression was prolonged even to the mice boosted with the same allergen 14 days after last treatment of OVA-DT. In addition, when the sensitized mice were boosted with the same allergens 14 days after last treatment of OVA-DT, the mice showed to increase the production of OVA-specific IgG2a/IgG3 and decreased that of OVA-specific IgE. OVA-DT targeted B cells bearing OVA-specific IgE, and killed them by DT-mediated cytotoxicity. CONCLUSION: The therapeutic effect of OVA-DT was specific to OVA-induced allergy and the suppression of OVA-induced allergy was continuously shown in the mice boosted with the same allergens. This is considered to be caused by the increase of OVA-specific IgG2a and IgG3, and because of the decrease of OVA-specific IgE by killing of B cells bearing OVA-specific IgE.

Unusual ferromagnetic couplings in single end-to-end azide-bridged cobalt(II) and nickel(II) chain systems.

Two new one-dimensional single azide-bridged metal(II) compounds [[M(5-methylpyrazole)4(N3)]n](ClO4)n(H2O)n [M = Co (1a), Ni (2a)] were prepared by treating an M(II) ion with stoichiometric amount of sodium azide in the presence of four equivalents of the 3(5)-methylpyrazole ligand. The isostructural compounds 1a and 2a crystallize in the monoclinic space group P2(1)/n. The azide bridging ligands have a unique end-to-end coordination mode that brings two neighboring metal centers into a cis-position with respect to the azide unit to form single end-to-end azide-bridged cobalt(II) and nickel(II) chains. The two neighboring metal atoms at inversion centers adopt octahedral environments with four equatorial 3(5)-methylpyrazole ligands and two axial azide bridges. Two adjacent equatorial least-squares planes form dihedral angles of 60.5 degrees and 60.6 degrees for Co and Ni, respectively. In addition, the metal-azide-metal units form large M-N3-M torsion angles, which are magnetically important geometrical parameters, of 71.6 degrees for M=Co and 75.7 degrees for M=Ni. It should also be noted that the M-N-N angles associated with end-to-end azide group, another magnetically important structural parameter, fall into the experimentally observed range of 120-140 degrees as 128.3(3) and 147.8(3) degrees for cobalt species and 128.4(2) and 146.1(3) degrees for nickel species; these values deviate from the theoretical value of around 164 degrees at which the incidental orthogonality is achieved under the torsion angle of 0 degrees. The compounds 1a and 2a have unique magnetic properties of ferromagnetism, zero-field splitting, and spin canting. The MO calculations indicate that the quasiorthogonality between the magnetic orbitals of metal ions and the p atomic orbitals of the bridging azide is possible in the observed structures and leads to the ferromagnetism. The spin canting related to the perturbation of ferromagnetism arises from the magnetic anisotropy and antisymmetric interactions judged by the structural parameters of the zero-field splitting and the tilted MN4 planes in a chain. The enhancement of magnetic interactions was accomplished by dehydrating the chain compounds to afford two soft magnets with critical temperature T(C) and coercive field of 2 K and 35 G for 1b and 2.3 K and 20 G for 2b, respectively.

Occupational asthma due to formaldehyde.

Formaldehyde is a low molecular weight chemical and can elicit acute and chronic health related problems. Most of the inhaled formaldehyde is retained in the upper respiratory tract due to its extraordinary solubility. Therefore, cases of formaldehyde-induced occupational asthma are sporadic despite its widespread use in industrial processes. We herein report upon a case of occupational asthma due to formaldehyde, which was confirmed by workplace challenge including working environmental assessments, and by formaldehyde inhalation challenge using a specially designed closed-circuit apparatus. To investigate the possible involvement of an IgE-mediated mechanism, both in vitro and in vivo tests were done. IgE antibody specific for formaldehyde-human serum albumin conjugate (F-HSA) was not detected by ELISA, and no specific cutaneous reactivity to F-HSA was noted by either skin prick or intradermal test. The patient was diagnosed with formaldehyde-induced occupational asthma not associated with an IgE mediated mechanism.

Role of skin prick test and serological measurement of specific IgE in the diagnosis of occupational asthma resulting from exposure to vinyl sulphone reactive dyes.

OBJECTIVES: Some patients with occupational asthma resulting from exposure to reactive dyes have skin reactivity to the causative dyes and specific IgE to reactive dyes have been found in these patients. However, the usefulness of skin prick tests (SPTs) and serological measurement of specific IgE in screening, diagnosis, and monitoring the occupational asthma resulting from exposure to reactive dyes have not yet been assessed. In this study, the clinical validation of SPTs and measurement of specific IgE to vinyl sulphone reactive dyes by enzyme linked immunosorbent assay (ELISA) was evaluated. METHODS: 42 Patients with occupational asthma from reactive dyes (true positive group) were enrolled. In these the causative reactive dye was confirmed by bronchial challenge test. 93 Asymptomatic factory workers with negative challenge to the reactive dye (true negative group) and 16 unexposed controls with negative challenge to the reactive dye were also enrolled. Skin prick tests were done with 10 mg/ml reactive dye in 0.4% phenol/0.9% saline. IgE specific to reactive dye conjugated to human serum albumin (HSA) was measured with enzyme linked immunosorbent assays (ELISAs). RESULTS: None of the unexposed controls had a positive response to SPTs. The sensitivity (76.2% v 53.7%), specificity (91.4% v 86.0%), positive predictive value (80.0% v 62.9%), and negative predictive value (89.5% v 80.8%) of SPTs were higher than those of ELISAs. The mean weal size of reaction to reactive dye was weakly correlated with the ELISA optical density of IgE to reactive dye conjugate in patients with occupational asthma from reactive dyes (n=41, r=0.337, p<0.05). In four patients with occupational asthma from reactive dyes and eight control subjects exposed to reactive dye, IgE specific to reactive dye conjugated to HSA was detected with ELISA even though they showed negative skin reactivity. Six patients completely avoided the reactive dye for a mean (SD) 27.8 (10.3) months, IgE specific to reactive dyes decreased in all six patients (p<0.05) during this time. CONCLUSIONS: Both SPTs and detection of IgE specific to reactive dye in serum samples could be valuable for screening, diagnosis, and monitoring occupational asthma resulting from exposure to reactive dyes. These two tests would complement each other.

Comparison of allergenic components between German cockroach whole body and fecal extracts.

BACKGROUND: Cockroaches have been demonstrated to be an etiologic factor in allergic diseases. Further, sensitivity to cockroach places patients with asthma at risk for exacerbations that require emergency medical care. OBJECTIVE: This study compared the differences in allergenic components between German cockroach whole body and German cockroach fecal extracts (GWBE and GFE). METHODS: Patients with asthma and/or allergic rhinitis were skin prick tested with German cockroach extract (Bayer Corporation, West Haven, CT). Serum specimens from these patients, 25 with positive skin tests and 8 with negative tests, were used for the ELISA and immunoblot experiments. RESULTS: By ELISA, 72% (18 of 25) and 60% (15 of 25) of positive responders' sera showed IgE antibodies to GWBE and GFE, respectively, and the IgE levels to GWBE were highly correlated with those to GFE (r = .84, P < .01). In inhibition ELISA experiments, extensive cross-reactivity was observed between GWBE and GFE, slight cross-reactivity between GWBE and Dermatophagoides farinae, and no cross-reactivity between GFE and D. farinae. The two-site monoclonal antibody ELISA detected more of the German cockroach major allergens in GFE compared with GWBE; 6.2 times (2420 vs 390 U/mL) for Bla g 1 and 3 times (15.32 vs 5.07 microg/mL) for Bla g 2. In the immunoblot comparison of patients' sera, the IgE antibodies binding to GWBE were apparently different from those binding to GFE in all the positive responders' sera; eg, 50% or more of the 25 positive responders' sera reacted to 43- to 67-kDa proteins in GWBE and to 28- to 30-kDa proteins in GFE, respectively. No IgE antibodies bound to components in GWBE and GFE in the 8 negative responders' sera. CONCLUSIONS: There are major differences between the allergenic components of GWBE and GFE. Based on the amounts of major allergens (Bla g 1, Bla g 2), German cockroach feces are a more important source of allergen than the whole body in respiratory allergic diseases.

Effects of gamma radiation on the conformational and antigenic properties of a heat-stable major allergen in brown shrimp.

This study was performed to evaluate the application of food irradiation technology as a method for reducing shrimp allergy without adverse effects. Shrimp heat-stable protein (HSP) was isolated and gamma irradiated at 0, 1, 3, 5, 7, or 10 kGy in the condition of solution (1 mg/ml), and fresh shrimp was also irradiated. Conformational change of irradiated HSP was monitored by means of spectrometric measures, enzyme-linked immunosorbent assay with mouse monoclonal antibody, or human patients' sera and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The ability of the immunoglobulin E of patients allergic to shrimp to bind to irradiated HSP was dose dependently reduced. The amount of intact HSP in an irradiated solution was reduced by gamma irradiation, depending on the dose. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the main band disappeared and the traces induced from coagulation appeared at a higher molecular weight zone. The binding ability of immunoglobulin E to allergens in the extracts from irradiated shrimp decreased, depending on the dose. The results provide a new method so that food irradiation technology can be applied to reduce allergenicity of shrimp.

Contrasting role of presenilin-1 and presenilin-2 in neuronal differentiation in vitro.

Presenilin-1 (PS1) and presenilin-2 (PS2), the major genes of familial Alzheimer's disease, are homologous to sel-12, a Caenorhabditis elegans gene involved in cell fate decision during development. Recently, wild-type and mutant presenilins have been associated also with apoptotic cell death. By using stable transfection of antisense cDNAs, we studied the functions of PS1 and PS2 during neuronal differentiation in the NTera2 human teratocarcinoma (NT2) cell line. Expression of antisense PS1 resulted in a failure of the clones to differentiate into neurons after retinoic acid induction, whereas cells transfected with antisense PS2 differentiated normally. Concomitantly, antisense PS1 clones were associated with increased apoptosis both under basal conditions and during the early period of neuronal differentiation after retinoic acid treatment. Overexpression of bcl-2 in antisense PS1 clones reduced cell death and resulted in a recovery of neuronal differentiation. These studies suggest that PS1 plays a role in differentiation and cell death and that PS1 and PS2 have differing physiological roles in this experimental paradigm.

Structural modeling of differential diagnosis, treatment, and results for allergic rhinitis.

This paper analyzed the relationship among the differential diagnosis, treatment, and results for allergic rhinitis using the covariance structural model. The data were collected from 274 patients with suspected allergic rhinitis who visited the Otorlaryngology Department of the Paik Hospital during 1991-1993. After each patient's characteristics was categorized and combined into several common factors, covariance structure analysis was performed to analyze the structural relationships among the differential diagnosis, treatment, and results of treatment using the significant factors obtained from discriminant analysis. The significant characteristics influenced the diagnosis were the results of skin test from mite/animal, and from mugworts, the results from laboratory tests, rhinorrhea and sneezing, and nasal polyps. The significant characteristics that influenced the method of treatment were: nasal polyps, headache/general symptom, family history/medication, and septal deviation. Headache/general symptom was the only significantly influencing factor for the treatment results.

Mono-ortho- and non-ortho-substituted polychlorinated biphenyls in human milk from Mohawk and control women: effects of maternal factors and previous lactation.

Fifty-four individual human milk samples from 50 mothers (20 Mohawks and 30 controls) were analyzed for four non-ortho- and eight mono-ortho-substituted polychlorinated biphenyls (PCBs). Mean total coplanar PCBs concentrations were 49 ng/g and 55 ng/g lipid for Mohawk and control women, respectively. A statistical evaluation of all analytical data reveals no significant difference of total coplanar PCB level between Mohawk and control women. The level of these contaminants is influenced by the age of the mother, number of breastfed children, and length of nursing period. Older women, primiparae, and cigarette smokers had higher levels of coplanar PCBs. In general, women had higher levels of coplanar PCBs in the first lactation and in the earlier samples of a given lactation, while levels declined both with duration of breast-feeding and with number of children nursed. The contribution of individual non-ortho- and mono-ortho-substituted PCB congeners to the total calculated toxic equivalent values (sigma TEQ) was assessed for the breast milk samples. The levels of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) in human milk of pooled specimens from Los Angeles, California and Binghamton, New York, widely separate cities in the United States (Schecter et al. 1989), were presented for reference purpose. The main contributions to the sigma TEQ were PCB congeners #118 (25.8 pg/g lipid), #126 (25 pg/g lipid), #105 (10.8 pg/g lipid), and #156 (7.4 pg/g lipid). Collectively, these compounds accounted for 70% of the sigma TEQ values.(ABSTRACT TRUNCATED AT 250 WORDS)

The significance of specific IgG and IgG4 antibodies to a reactive dye in exposed workers.

To evaluate the significance of specific IgG and specific IgG4 in the development of work-related respiratory symptoms, specific IgG and specific IgG4 to Black GR-human serum albumin (HSA) conjugate were measured by ELISA in 309 dye-exposed workers and 63 unexposed patients as negative controls. A survey revealed that 78 (25.2%) had work-related lower respiratory symptoms with or without nasal, skin or eye symptoms. Specific IgG and specific IgG4 were detected in 23% and 14% of the exposed workers, respectively. The prevalence of specific IgG and specific IgG4 was significantly higher in smokers and workers with specific IgE or those with lower respiratory symptoms (P less than 0.05), but was not associated with work station, duration of dye exposure or atopy. These results suggested that the existence of specific IgG to Black GR-HSA might represent a response to Black GR exposure and be closely related with work-related respiratory symptoms.

Clinical and immunologic evaluations of reactive dye-exposed workers.

To evaluate type 1 hypersensitivity to reactive dyes, its prevalence, and its relationship to respiratory dysfunction, we studied clinical and immunologic features, including skin prick tests. RAST, and bronchoprovocation tests, of 309 employees working in a reactive-dye industry. Our survey revealed that 78 (25.2%) employees had work-related lower respiratory symptoms associated with or without nasal, skin, or eye symptoms. Among these employees, 38 (48.7%) had nonspecific bronchial reactivity. Thirteen demonstrated immediate (6), dual (6), or late only (1) asthmatic responses after inhalation of four kinds of reactive-dye solutions. Twenty-five employees demonstrated immediate skin responses to black GR dye, and 21 reacted to orange 3R. Fifty-three employees (17%) had specific serum IgE antibody against black GR and orange 3R-human serum albumin conjugate. Specific IgE was detected more frequently in symptomatic employees (30%) and smokers (100%). No association was found between atopy and specific IgE binding. The RAST-inhibition tests of black GR revealed significant inhibitions by black GR-human serum albumin conjugate and minimal inhibitions by unconjugated black GR. Orange 3R RAST-inhibition tests revealed significant inhibitions by conjugated forms of black GR and orange 3R and some inhibitions by two unconjugated dyes, suggesting an immunologic cross-reactivity between these dyes. These findings suggested that reactive dyes could induce immunologic responses, most likely IgE-mediated.