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At least 19 recordsLinked to original sources

Identification of carmine allergens among three carmine allergy patients.

BACKGROUND: There have been several reports of carmine allergy; however, identification of the responsible carmine allergens has not been widely documented. METHODS: Three female patients presented with a history of anaphylaxis and/or urticaria/angioedema after ingestion of carmine-containing foods. All three patients had 4+ skin prick tests to carmine. Among them, two patients were confirmed to have carmine allergy by blinded, placebo-controlled food challenges to carmine. SDS-PAGE of cochineal insects and carmine, immunoblotting for IgE antibody with sera from all three patients, and immunoblotting inhibition with carmine were performed. RESULTS: SDS PAGE of minced cochineal insects revealed several protein bands of 23-88 kDa. Several of these bands were variably recognized by our three patients' sera, and this reactivity was inhibited by carmine. Although no protein bands could be visualized on SDS-PAGE of carmine in Coomassie brilliant blue staining, three protein bands were recognized by two of the three patients' serum. CONCLUSIONS: These results suggest that commercial carmine retains protein-aceous material from the source insects. These insect-derived proteins (possibly complexed with carminic acid) are responsible for IgE-mediated carmine allergy. Patient reactivity to these proteins may vary.

Administration, Oral↗

High-performance liquid chromatographic separation of carminic acid, alpha- and beta-bixin, and alpha- and beta-norbixin, and the determination of carminic acid in foods.

During a study of natural food colours, a simple and reliable high-performance liquid chromatography system was developed for use with cochineal and annato. An isocratic mobile phase, consisting of methanol and 6% aqueous acetic acid, resolved bixin and norbixin, while a gradient system was used to separate carminic acid and the annato compounds. The carminic acid contents of cochineal extract, carmine and carmine hydrosoluble were determined using an isocratic mobile phase (40:60, v/v). The detection limit for carminic acid in the various products was approximately 100 ng/g. Carminic acid was determined quantitatively in fruit beverages, yogurt and candies. It was demonstrated that, because of decomposition, carminic acid was not suitable for use in candies when manufacturing temperatures above 100 degrees C were required. Most membrane filters are not suitable for use with cochineal solutions, but a cellulose membrane filter did not adsorb carminic acid and was used successfully to remove impurities from water-based cochineal products and food extracts containing carminic acid.

Carmine↗

A method for determining identity and relative purity of carmine, carminic acid and aminocarminic acid.

Carmine is one of the few dyes currently certified by the Biological Stain Commission that is not assayed for dye content. Existing assay methods are complex and do not differentiate the three cochineal derivatives carmine, carminic acid and aminocarminic acid. The latter dye is relatively new to the food trade as an acid-stable red colorant and may eventually enter the biological stains market. The assay proposed here is a two-step procedure using quantitative spectrophotometric analysis at high pH (12.5-12.6) followed by a qualitative scan of a low pH (1.90-2.10) solution. Carmine is distinct at high pH, and the remaining dyes are easily distinguished at low pH. Four instances of mislabeling are documented from 18 commercial products, but the mislabeled dyes were not certified dyes. Samples from nearly all lots of carmine certified by the Biological Stain Commission from 1920 to 2004 proved to be carmine, but they varied widely in dye content. Batches from 1920 through the 1940s were significantly richer in dye content. Variability has been extreme since 2000, and most of the poorest lots have been submitted since 1990.

Biological Assay↗

Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins.

Indigo carmine blue is a water-soluble, light-sensitive, anionic dye most widely applied in microscopic staining techniques. It is virtually nonfluorescent in its oxidized state in aqueous solution, but when reduced under alkaline conditions, it becomes fluorescent, with absorption and emission maxima at 436 and 528 nm, respectively. It is demonstrated that the fluorescent character of the reduced form of the dye can be exploited as a label for the determination of cationic proteins by capillary electrophoresis with laser-induced fluorescence detection. Model proteins trypsinogen and cytochrome c are noncovalently labeled with indigo carmine; the bound species are reduced, rendering the indigo carmine fluorescent; and capillary electrophoresis with laser-induced fluorescence detection is used for subsequent analysis. Various buffer systems and buffer additives were examined and optimized, and a suitable pH range for optimal fluorescence intensity and protein-dye interaction was established. Fluorescence quenching of the reduced dye when bound to protein was observed in all buffer systems.

Coloring Agents↗

Indigo carmine inhibits endothelium-dependent and -independent vasodilation.

To investigate the potential mechanisms by which indigo carmine produces hypertension, we tested the hypothesis that indigo carmine inhibits endothelium-dependent vasodilation and determined the possible site of the inhibition (endothelium versus smooth muscle). Using isolated rat thoracic aortic rings that were precontracted with phenylephrine, we examined vasodilatory responses to acetylcholine, histamine, and Ca2+ ionophore A23187 (in endothelium-intact rings) and sodium nitroprusside and isoproterenol (in endothelium-denuded rings) in the presence and absence of indigo carmine. In addition, the effects of methylene blue on the acetylcholine- and sodium nitroprusside-induced vasodilation were compared with those of indigo carmine. Indigo carmine (10(-6), 10(-5), and 10(-4) mol/L) significantly inhibited receptor- and non-receptor-mediated endothelium-dependent vasorelaxation. Indigo carmine (10(-4) mol/L) also inhibited endothelium-independent vasorelaxation induced by sodium nitroprusside (an activator of vascular smooth muscle soluble guanylyl cyclase), although to a lesser extent than vasodilation from acetylcholine, histamine, and Ca2+ ionophore A23187. In contrast, indigo carmine (10(-4) mol/L) had no effect on the vasodilation induced by isoproterenol (an activator of adenylyl cyclase), indicating that indigo carmine selectively inhibits nitric oxide-mediated responses. Methylene blue, a known inhibitor of soluble guanylyl cyclase, inhibited both acetylcholine- and sodium nitroprusside-induced vasorelaxation. The inhibition was also greater in the acetylcholine- than the sodium nitroprusside-induced vasodilation. These results suggest that indigo carmine, like methylene blue, may inhibit endothelium-dependent relaxation by a mechanism that involves two levels. The major action of indigo carmine appears to be at the level of nitric oxide generation and/or release from the endothelial cell. In addition, indigo carmine appears to inhibit vascular smooth muscle guanylyl cyclase. Thus, indigo carmine may elevate blood pressure by interfering with these nitric oxide-mediated vasodilatory mechanisms.

Acetylcholine↗

Popsicle-induced anaphylaxis due to carmine dye allergy.

BACKGROUND: IgE-mediated hypersensitivity is a suggested mechanism to explain adverse reactions from carmine-containing products. OBJECTIVE: To describe a patient who experienced anaphylaxis after ingestion of a popsicle colored with carmine and to provide additional evidence that the adverse reaction was IgE-mediated. METHODS: The patient and her husband underwent skin prick tests to the popsicle and carmine. The patient also received skin prick tests and/or open oral challenge to each of the other components of the incriminated food. Topical application of cosmetics with and without carmine to the patient's forearm was also performed. To confirm carmine-specific IgE, a Prausnitz-Kustner (P-K) test was performed using the patient's husband as recipient. Twenty control subjects also were tested to carmine by skin prick test. RESULTS: The patient showed 4+ skin prick test responses to the popsicle and carmine. Skin prick tests and/or open oral challenge to each of the other components of the popsicle were negative. The patient's husband's and 20 control subjects' skin prick tests to carmine were negative as was the patient's husband's skin prick test to the popsicle. Skin prick test reactivity to the popsicle and carmine were successfully transferred to the patient's husband in P-K format. Cosmetics applied to the patient's forearm elicited no immediate response. CONCLUSION: The positive skin prick tests to the popsicle and carmine and the successful (P-K) transfer of skin prick test reactivity support a carmine-specific, IgE-mediated mechanism in explaining our patient's popsicle-induced anaphylaxis.

Adult↗

Carmine (E-120)-induced occupational asthma revisited.

BACKGROUND: Occupational asthma (OA) caused by carmine (E-120) has been reported. OBJECTIVE: We sought to evaluate the prevalence of sensitization and OA at a natural dye processing factory in which 2 workers had been given a diagnosis of carmine-induced OA 6 years previously. METHODS: The 24 current employees and one worker who had recently left work because of asthma completed a questionnaire and underwent skin testing (carmine, cochineal, carminic acid, curcuma, annato, and chlorophyll), carmine IgE dot-blot analysis, and methacholine inhalation testing. Workers exhibiting positive occupational skin test responses, work-related asthma, or bronchial hyperresponsiveness underwent specific inhalation challenge and serial peak expiratory flow rate recording. RESULTS: Positive skin test responses to carmine (41.7%), cochineal (29.2%), and carminic acid (4.2%) were observed. Carmine IgE dot-blot results were positive in 4 subjects. No difference in atopy or smoking was observed between occupationally sensitized and nonsensitized subjects. Among the 5 employees reporting work-related asthma, 2 had positive skin test responses, and 4 had bronchial hyperresponsiveness. Five subjects underwent specific inhalation challenges: 2 workers had early asthma responses to carmine and cochineal challenges, and the remaining subjects did not have suggestive peak expiratory flow recordings. The subject who had left his job was given a diagnosis of carmine-induced OA. CONCLUSION: The prevalence of sensitization and OA caused by carmine was 41.6% and 8.3%, respectively. When the 3 workers who had left their jobs were included, the cumulative incidence of sensitization and OA was 48.1% and 18.5%, resembling the healthy worker effect. Prevention programs to establish the permissible levels of airborne allergen should be implemented.

Adolescent↗

Occupational asthma to carmine in a butcher.

UNLABELLED: Hypersensitivity to carmine (E120) has been identified as a cause of food intolerance and occupational asthma. We present a case of occupational asthma following exposure to carmine in a manufacturer of sausages and review the literature. CASE REPORT: A 42-year-old non-atopic male presented with a 5-year history of rhinoconjunctivitis and asthma on occupational exposure to food additive dusts. Symptoms increased after work. The patient had been exposed for more than 20 years. METHODS: Skin prick tests were performed with a battery of common inhalant allergens and spices. Cochineal, carmine lake and additive mixes used by the patient were extracted and subsequently used for skin prick test, bronchial provocation and in vitro measurements (specific IgE, Western blot and chromatographic fractionation). RESULTS: Prick tests were positive to carmine and carmine-containing additives; carmine-specific IgE and bronchial challenge tests were also positive (PC20 = 0.0004 mg/ml and 1.6 kU/l). Western blot showed IgE binding to bands of about 30 kDa on cochineal extract and a diffuse pattern at 40-97 kDa on carmine. This result was confirmed by gel filtration chromatography and dot blot. Carmine completely inhibited IgE binding to cochineal extract. DISCUSSION: Carmine is a potential sensitizer in an occupational setting: 18 cases of occupational asthma have been described to date. Carmine allergens are poorly defined; in general, proteins from cochineal not removed by the extraction process are considered as the main allergens in carmine. Our results are consistent with this, but show that these proteins may be subject to chemical modification.

Adult↗

[Efficacy of contrast chromoendoscopy of the colon after oral administration of indigo carmine dye].

INTRODUCTION: Indigo carmine dye is usually spread directly over the colon in many chromoscopic techniques aiming better visualization of a lesion already detected by conventional colonoscopy. Examination of the colon already stained by oral administration of indigo carmine dye may increase detection of small lesions resulting in higher sensibility of the colonoscopy in diagnosing diminutive lesions. OBJECTIVE: Analyze the results regarding the quality of chromoscopic technique and the indigo carmine dye distribution over the colon after oral administration. PATIENTS AND METHODS: Fifty patients undergoing colonoscopy were evaluated. A capsule containing 100 mg of indigo carmine dye was offered to these patients 30 min before oral mannitol prep routinely used. The indigo carmine dye contrast effect was graded as bad, regular or good according to preestablished criteria in three segments of the colon: right and left colon and the rectum. RESULTS: In the right colon, good indigo carmine dye contrast effect was observed in only 9 (18.8%) patients, while it was considered regular and bad in 32 (66.6%) and in 7 (14.6%) patients, respectively. A good indigo carmine dye contrast effect was never observed in this series for the left colon or in the rectum. As a matter of fact, no indigo carmine dye was observed in the left colon in 80.9% and in the rectum in 92% of patients in this series. CONCLUSION: Although it may be simple and desirable, oral administration of indigo carmine dye seems ineffective for enhancing detection of diminutive lesions by chromoscopy as result of poor colonic distribution of indigo carmine dye mainly at distal colonic sites.

Administration, Oral↗

Occupational asthma and immunologic responses induced by inhaled carmine among employees at a factory making natural dyes.

Carmine is a natural red dye widely used as a food coloring agent and for cosmetic manufacture. It is extracted from the dried females of the insect Dactylopius coccus var. Costa (cochineal). Although it has been reported that inhalation of carmine may give rise to occupational asthma and extrinsic allergic alveolitis, there is little evidence of its immunogenic capacity. We studied nine current employees at a factory making natural dyes and one former employee who had left this plant after occupational asthma developed. A current employee had work-related symptoms of rhinitis and asthma that were confirmed by bronchial provocation tests, and another worker had rhinitis. Immunologic sensitization to carmine and cochineal was evaluated by means of skin testing and determination of serum-specific IgE and IgG subclass antibodies by RAST and ELISA, respectively. The specificity of the RAST assay was investigated by RAST inhibition with different fractions of carmine. The three workers with respiratory symptoms had positive skin prick test reactions to both carmine and cochineal. An immediate response to the bronchial provocation test with carmine and cochineal was observed in the current employee with asthma. Specific IgE antibodies against carmine and cochineal were found only in this worker. RAST inhibition studies indicated that the main allergen had a molecular weight between 10 and 30 kd. Specific IgG antibodies against carmine and cochineal, mainly the subclasses IgG1, IgG3, and IgG4, were found in the 10 subjects surveyed. These findings suggest that carmine may induce immunologic responses, most likely IgE mediated in workers with symptoms of occupational asthma.

Adolescent↗

Identification of allergens involved in occupational asthma due to carmine dye.

BACKGROUND: Carmine has been implicated as an etiologic agent of occupational asthma, but the allergens involved have not been yet identified. OBJECTIVE: To identify the allergens involved in occupational asthma due to carmine dye. METHODS: An in vitro study based in SDS-PAGE and IgE immunoblotting with carmine and cochineal extracts was performed. Sera from three carmine dye workers diagnosed with occupational asthma induced by carmine dye and from seven nonatopic subjects were used. RESULTS: Three proteins of around 30, 28, and 17 kD in raw cochineal extract and another protein of 50 kD in the boiled one were demonstrated by SDS-PAGE. Two proteins of around 50 and 28 kD were observed in the carmine extract by the same technique. Specific IgE binding bands at 17 kD in cochineal raw extract, at 50 kD in the boiled one, and at 28 kD in carmine extract were demonstrated by IgE immunoblotting. CONCLUSIONS: We have identified three allergens of around 17, 28, and 50 kD implicated in occupational asthma of three carmine workers.

Asthma↗

Occupational asthma induced by inhaled carmine among butchers.

BACKGROUND: Carmine is a natural coloring agent for food and cosmetics. There have been several reports of occupational asthma among employees at factories making natural dyes, however, there are no cases reported among butchers. OBJECTIVES: We report on two male patients who presented with a history of occupational asthma. Both patients are butchers and used a mixture of additives with carmine as dye in sausages. METHODS: Skin prick tests were performed with common aeroallergens, commercial mixture additives and separated compound of the additive mixture. Total IgE, metacholine test, specific inhalatory challenge tests with carmine and mites, SDS-PAGE and IgE-immunoblotting were carried out. RESULTS: Skin tests were positive to mites, additive mixture and carmine in both cases. Specific IgE to mites was positive only in patient one. Bronchial provocation tests were positive to metacholine and carmine and negative to Dermatophagoides pteronyssinus in both cases. IgE-immunoblot showed a specific IgE-binding band at 10 kDa and at a large number of bands along all molecular weights. No inhibition was achieved in carmine-IgE-specific bands with the D. pteronyssinus extract. CONCLUSIONS: We report on two butchers with occupational asthma due to an IgE-mediated sensitization to carmine. There are several proteins that may act as allergens, and these may be conditioned by the route of exposure and are not cross-reactive with mite proteins. Carmine may be considered to be an etiologic agent in butcher's asthma.

Adult↗

[Asthma and allergy due to carmine dye].

Cochineal carmine, or simply carmine (E120), is a red colouring that is obtained from the dried bodies of the female insect Dactylopius coccus Costa (the cochineal insect). We have evaluated the prevalence of sensitization and asthma caused by carmine in a factory using natural colouring, following the diagnosis of two workers with occupational asthma. The accumulated incidence of sensitization and occupational asthma due to carmine in this factory are 48.1% and 18.5% respectively, figures that make the introduction of preventive measures obligatory. Occupational asthma caused by inhaling carmine should be considered as a further example of the capacity of certain protein particles of arthropods (in this case cochineal insects) to act as aeroallergens. Carmine should be added to the list of agents capable of producing occupational asthma, whose mechanism, according to our studies, would be immunological mediated by IgE antibodies in the face of diverse allergens of high molecular weight, which can vary from patient to patient. Nonetheless, given the existence of different components in carmine, it cannot be ruled out that substances of low molecular weight, such as carminic acid, might act as haptenes. Besides, since we are dealing with a colouring that is widely used as a food additive, as a pharmaceutical excipient and in the composition of numerous cosmetics, it is not surprising that allergic reactions can appear both through ingestion and through direct cutaneous contact. We find ourselves facing a new example of an allergen that can act through both inhalation and digestion, giving rise to an allergolical syndrome that can show itself clinically with expressions of both respiratory allergy and alimentary allergy.

Allergens↗

Enzyme immunoassay for carminic acid in foods.

A competitive enzyme immunoassay (EIA) for carminic acid was investigated. Monoclonal anticarminic acid antibody was obtained from A/J mice immunized with carminic acid-human immunoglobulin G (IgG) conjugate. Carminic acid was extracted with distilled water from beverage, jelly, candy, pasta sauce, yogurt, or ice cream samples. Ham or fish paste samples were digested with pronase, then carminic acid was extracted from samples with sodium hydroxide solution. The extract was diluted more than 10-fold with 1% gelatin in borate buffer solution. Microtiter plates were coated with carminic acid-bovine serum albumin (BSA) conjugate or just BSA. Goat anti-mouse IgG(H+L)-peroxidase complex was used as a second antibody, and 3,3',5,5'-tetramethylbenzidine was used as a substrate for the peroxidase. The working range for quantitative analysis was 0.3-10 ng/mL, and the detection limit was 0.2 micrograms/g original sample. Recoveries of carminic acid by this assay were > 95% for milk beverage and jelly, and > 85% for yogurt and fish paste. Carminic acid was detected in 7 of 26 red-colored commercial food products and ranged from 3.5 to 356 micrograms/g. This EIA system also responded to the structural analogue of carminic acid, laccaic acid.

Animals↗