Occupational asthma and food allergy due to carmine

Carmine (E120), a natural red dye extracted from the dried females of the insect Dactylopius coccus var. Costa (eochineal), has been reported to cause hypersensitivity reactions. We report a case of occupational asthma and food allergy due to carmine in a worker not engaged in dye manufacturing. A 35-year-old nonatopic man, who had worked for 4 years in a spice warehouse, reported asthma and rhinoeonjunctivitis for 5 months, related to carmine handling in his work. Two weeks before the visit, he reported one similar episode after the ingestion of a red-colored sweet containing carmine. Peak flow showed drops higher than 25% related to carmine exposure. Prick tests with the cochineal insect and carmine were positive, but negative to common aeroallergens, several mites, foods, and spices. The methacholine test was positive. Specific bronchial challenge test with a cochineal extract was positive with a dual pattern (20% and 24% fall in FEV_i). Double-blind oral challenge with E120 was positive. The patient's sera contained specific IgE for various high-molecular-weight proteins from the cochineal extract, as shown by immunoblotting. Carmine proteins can induce IgE-mediated food allergy and occupational asthma in workers using products where its presence could be easily overlooked, as well as in dye manufacture workers.     

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.     

Occupational asthma to carmine in a butcher

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.     

醋酸-靛胭脂染色在基层医院早期胃癌及癌前病变诊断中的应用

目的探讨提高基层医院早期胃癌及癌前病变检出率的方法。方法经胃镜检查发现胃黏膜异常进行醋酸-靛胭脂染色后取活检病理检查患者72例为染色组,同期胃镜下发现黏膜异常患者直接取组织活检病理检查患者68例为对照组,观察胃黏膜染色情况并与病理检查结果对比分析,比较2组患者早期胃癌及癌前病变的检出率。结果醋酸-靛胭脂染色后胃黏膜表现为黏膜褪色(16.7%)、着色不良(63.9%)及着色均匀(14.3%)。其中染色组黏膜褪色患者中早期胃癌或高级别上皮内瘤变的检出率(91.7%)显著高于着色不良(8.6%)和着色均匀(0.0%);黏膜着色不良中低级别上皮内瘤变或肠上皮化生比例(82.6%)显著高于黏膜褪色(8.3%)和着色均匀(14.3%)。染色组早期胃癌及癌前病变检出率(13.9%,63.9%)均分别显著高于对照组(2.9%,29.4%)。结论胃镜下醋酸-靛胭脂染色能够提高基层医院早期胃癌及癌前病变的检出率,并且成本低廉,操作简便,适宜于基层医院推广应用。     

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.     

Vascular volume determination of articular tissues in normal and anterior cruciate ligament-deficient rabbit knees

The vasculature of diarthroidal joints has been well documented; however, the volume of vessels supplying different articular tissues is unknown. Angiogenesis, the formation of new vessels from preexisting ones, is difficult to quantify in joints due to the unavailability of a suitable technique. Although angiogenesis is known to occur in rheumatoid arthritis, the development of new vessels following joint injury has not been ascertained. A vascular casting technique was developed using carmine red dye to measure the vascular volume of the medial collateral ligament (MCL), lateral collateral ligament (LCL), menisci, medial capsule, and infrapatellar fat pad of the rabbit knee joint. Vascular volume determinations were repeated at 4 weeks in a group of anterior cruciate ligament (ACL)-transected animals and in a sham-operated control group. The volume of vessels supplying the MCL was estimated to be 0.22 ± 0.07 μl (mean ± S.E.M.), the LCL volume was 0.25 ± 0.05 μl, the medial meniscus volume was 0.19 ± 0.03 μl, the lateral meniscus volume was 0.40 ± 0.08 μl, the medial capsule volume was 0.14 ± 0.05 μl, and the infrapatellar fat pad volume was 1.90 ± 0.62 μl. Following ACL transection, angiogenesis was found to occur in the MCL only. All other tissue vascularities were not significantly different from sham-operated controls. A quantifiable method for measuring vascular volume of knee joint tissues has been described. Joint instability stimulates angiogenesis in the ipsilateral MCL; however, the absence of angiogenic activity in other articular tissues might help explain the lack of posttraumatic healing associated with these joints. Anat. Rec. 251:207–213, 1998. © 1998 Wiley-Liss, Inc.     

Occupational asthma due to inhaled carmine

Two patients are described with occupational asthma due to carmine, a natural dye extracted from the insect Coccus cactus. Both had dual asthmatic reactons after carmine inhalation. Oral challenge provoked gastrointestinal symptoms in one patient, and asthma in them both, perhaps accounting for their continuing symptoms. One patient worked extracting carmine from the insects and the other used carmine as a cosmetic colouring agent.     

Murine acinar pancreatitis preceding necrotizing myocarditis after Coxsackievirus B3 inoculation.

Balb/C weanling mice were inoculated intraperitoneally with a myocarditic variant of coxsackie-virus B3, with the aim of characterizing more fully the cell damage induced in the heart as well as in other organs. During the first week postinfection (pi), all animals developed acinar pancreatitis, followed by focal myocarditis. In accordance with the increasing infectivity titers, such progressive histopathological changes correlated with local viral replication. From day 4 pi, acinar degeneration accompanied by diffuse inflammatory exudate was observed in the pancreas, followed by fatty tissue replacement by day 8. In the heart, focal necrosis rather than inflammatory reaction first appeared at 4 days pi and became widespread by 6-8 days pi. Necrotic foci usually presented calcium deposits, with absence of myofibrils in the affected fibers. The fact that both periodic acid Schiff (PAS) and Best carmine staining remained positive even after diastase treatment ruled out basophilic necrosis. In summary, the pancreas appeared to be the site of primary viral replication leading to viremia.     

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.     

Histotopochemical investigations on elastic membranes of blood vessels with special regard to diabetes mellitus. II. Carbohydrates (author's transl)]

Investigations on carbonhydrates were carried out with special regard to acid mucopolysaccharides and glycogen. The fluorescence-microscopical proof of acid micropolysaccharides with acridinorange (pH=3,3) and by means of the pseudoisocyanin-reaction (proof of-SO3H-groups) gave positive results on elastic membranes of blood vessels. These results were correlating to the increase of the diabetes mellitus. Healthy Wistar-rats did not show metachromasia with toluidinblue in the walls of the blood vessels, whereas in streptozotocin-diabetes there was strong metachromasia shown by these structures. The stages of diabetes in man were also correlated to an increase of metachromasia. The PAS-reaction, the staining with Best's carmine and the reaction with alizarinblue S for the proof of glycogen were positive in all blood vessels investigated.     

Indigo carmine for the selective endoscopic intervertebral nuclectomy

This study was undertaken to prove that the selectively infiltrated parts of nucleus pulposus with indigo carmine was degenerated parts of nucleus pulposus. This study was done, between August and October 2002, in 5 patients, who received endoscopic discectomy, due to intervertebral disc herniation. Discogram was done with mixture of indigo carmine and radioactive dye. Blue discolored part was removed through endoscope, and small undiscolored part was removed together for the control. The two parts were stained with hematoxylin and eosin and compared under the microscope. Undiscolored part was normal nucleus pulposus, composed of chondrocytes with a matrix of type II collagen and proteoglycan, mainly aggrecan. However, in discolored part, slits with destruction of collagen fiber array and ingrowth of vessel and nerve were observed. Using indigo carmine in endoscopic discectomy gives us selective removal of degenerated disc.     

On the problem of the excretory function of the placental epithelium

Summary In the experiments on rabbits and white rats it has been shown that the dye (indigo carmine injected into the blood of an embryo) is secreted by the syncytiotrophoblast of the chorion of the placenta into the blood of the maternal organism. Indigo carmine has been also found in the epithelium of the vitalline placenta which functions for a long period of time in rodents. By means of the xanthydrol reaction crystals of the dixanthylurea were always found in the chorionic syncitiotrophoblast. The author draws the conclusion that it is chiefly the chorionic syncitiotrophoblast which performs the excretion of the metabolic products of the embryo.Submitted by Active Member of the Academy of Medical Sciences of the USSR, V. N. Ternovsky     

Histomorphological and histochemical study of mucosa of fallopian tube of ageing goat.

The mucosa of infundibulum, ampulla and isthmus of fallopian tubes from 24 female goats aged from 2 months 24 days to 5 years 2 months and 18 days have been studied. The transition of mucosal folds, the lining epithelium and its cell types from Infundibulum towards uterine end, and changes in relation to the age and phases of estrous cycle have been reported. The most striking findings were: a) in goat the fallopian tube like that of sheep was lined chiefly by nonciliated type of cells in infundibulum, whereas the ciliated cells increased caudally, almost all being nonsecretory type in the isthmus; b) the presence of smooth muscle cells as identified with Gomori's one step trichrome stain was observed in the goat fallopian tube in case of mucosal folds specifically at the sites of branchings which anastomose with the adjacent ones; c) the PAS positive secretory material was found to be associated with glycogen acid mucopolysaccharides and mucoproteins as demonstrated by Best's carmine, Alcian blue-PAS and Bismark brown reactions, respectively.     

卡红染色法在鸡胚早期血管发育研究中的应用

目的探讨卡红染色在鸡胚原肠期血管发育研究中的应用。方法种蛋置于恒温培养箱进行孵化,分别取不同时段鸡原肠胚,经4%多聚甲醛固定,硼砂卡红溶液染色后,体式显微镜下观察。结果鸡原肠期的胚体及外周的血岛呈鲜红色,可相继观察到原条、脊索、神经管及体节的发生过程,血岛的出现及其变化过程。结论能更早、更直观地观察到鸡胚早期血管发生的变化过程;此法简单易行,可用于对胚胎发育及血管发生等影响因子的筛选和研究。     

Flat depressed early colon cancer--a case report.

A flat depressed early colon cancer (FDEC) is characterized by non-polypoid growth pattern, no association of adenomatous tissues and a tendency of even small lesions toward submucosal invasion and lymph node metastasis. It supports de novo carcinogenesis of colorectal cancer, although most colorectal cancers arise in pre-existing adenoma (adenoma-carcinoma sequence). There have been few reports of small depressed cancers because of the difficulty in colonoscopic detection and the rapid development to ulcerating advanced cancers. We report a case of flat depressed early colon cancer confined to mucosa detected by indigo carmine contrast colonoscopy.     

Influence of levocabastine suspension on ciliary beat frequency and mucociliary clearance

The effects of levocabastine, a new fast-acting, highly potent H1-antagonist, on nasal ciliary epithelial function were investigated in an in vitro and in vivo study. In the in vitro study, a suspension of levocabastine in Locke-Ringer solution was applied to 10 bioptic specimens of ciliated human adenoid tissue. Each specimen was exposed to the test solution for 60 min. Ciliary beat frequency (CBF) was recorded with a photoelectric recording device at 10-min intervals. There were small, insignificant decreases in CBF, which were minimal compared to that observed with ciliotoxic agents. In the in vivo study, 8 healthy volunteers were given, intranasally, one droplet of the levocabastine suspension. Mucociliary transit time (MTT) was measured by placing a saccharin particle drenched in indigo carmine in the nose just below the top of the concha and measuring the time until appearance of the dye in the pharyngeal cavity. No statistically significant differences were found in the MTT before and after application of the levocabastine suspension. The studies thus indicate that nasally administered levocabastine does not interfere with ciliary beat frequency and mucociliary function.     

Phenotypical and morphological alterations to rat sinusoidal endothelial cells in arterialized livers after portal branch ligation

The hepatic sinusoids are preferentially supplied with portal venous blood and equipped with fenestrated endothelial cells that are distinct from capillary endothelial cells. We previously observed in rats that sinusoidal capillarization proceeded concurrently with arterial blood supply during hepatocarcinogenesis. This study aimed to clarify the inducing role of arterialization in sinusoidal capillarization by investigating phenotypical, morphological and functional alterations to sinusoidal endothelial cells (SECs) in arterialized rat livers induced by portal branch ligation. At one week, after massive hepatic necrosis following ligation, the livers were restored to their normal architecture without causing post-necrotic fibrosis. At 12-21 weeks, they exhibited a normal histology except for mild pericellular fibrosis which developed along sinusoids or between adjacent hepatocytes. SECs expressed factor VIII-related antigen and showed a decrease in the number of fenestrae and porosity, still lacking any basement membrane but further retaining the functional capacity for carmine dye uptake. Stellate cells, while occasionally associated with large amounts of collagen bundles, contained many lipid droplets and expressed no alpha-smooth muscle actin, indicating a quiescent property. Kupffer cells were commonly found within the sinusoids. The present results indicate that arterialization of the liver induces a partial (but not complete) transition of SECs into capillary-type endothelial cells, suggesting that arterialization might be one of the factors which induce sinusoidal capillarization in the development of hepatocellular carcinoma.     

Late asthmatic reactions and changes in histamine responsiveness provoked by occupational agents

The temporal and quantitative relationship between increases in airway responsiveness and late asthmatic reactions provoked by inhalation challenge with occupational agents was studied in nine individuals who underwent a total of thirteen active inhalation challenge tests with one of the following agents: toluene diisocyanate (TDI), maleic anhydride (MA), trimellitic anhydride (TMA), carmine, or colophony (pine wood resin). Airway responsiveness to inhaled histamine (histamine PC20) was measured before and at approximately 3 and 24 h after control and active challenge exposure, when, on all but four occasions, FEV1 was within 10% of pre-challenge values. Significant increases (p less than 0.02) in histamine responsiveness were present at 3 h following challenge exposures which subsequently provoked a definite late asthmatic reaction (FEV1 decrease greater than 15% 3-11 h post challenge). These increases in histamine responsiveness were significantly greater than those at 3 h following the challenges which provoked an isolated early (FEV1 decrease less than 6% 3-11 h post-challenge) or equivocal late asthmatic reaction (FEV1 decrease 6-15% 3-11 h post-challenge) (p less than 0.03). Although histamine responsiveness remained high at 24 h after challenges provoking late asthmatic reactions (p less than 0.05), this was less than the increase at 3 h and not significantly different from the PC20 at 24 h after challenges provoking either single early or equivocal late asthmatic reactions.(ABSTRACT TRUNCATED AT 250 WORDS)     

Nonspecific esterase activity in malignant non-lymphogranulomatous lymphomas]

It is recommended to determine the activity of nonspecific esterase in malignant non-lymphogranulomatous lymphomas (MNL) and to use for this purpose 5-bromindoxylacetate with poststaining of sections with aluminous carmine. In lymph nodes, affected with MNL, this procedure facilitates the detection of the arrangement of reticular cells and the degree of MNL penetration with hystiocytes. Also such structural features of MNL as their follicular structure and infiltration of sinuses of lymph nodes with the tumor tissue are determined.     

Staining of Fasciola hepatica by natural herbal dyes

The plant kingdom is a valuable source of new medicinal agents. Traditional synthetic dyes used for staining worms may have some carcinogenic effects; therefore, the use of herbal dyes as an effective alternative may be interesting. The objective of the current study was to evaluate the staining effect of herbal dyes on Fasciola hepatica. Powdered alizarin, henna and curcuma in concentrations of 5, 10 and 20 g/100 mL of distilled water were used to stain a F. hepatica worm according to the carmine staining method. Our criteria for evaluation were the extent of staining on different parts of the worm such as suckers, intestine, testes, spines, ootype, vitelline duct, vitellaria, ovary, uterus and eggs. The results showed that alizarin (10 g/100 mL; 10%, w/v) was a better dye than henna (20 g/100 mL; 20%, w/v) and curcuma (20 g/100 mL; 20%, w/v) for the staining of suckers, branched intestine, vitelline duct and ootype. Although henna stained the whole body of the worm better than curcuma, curcuma had a better staining effect on surface spines, eggs and especially branched testes. In comparison with carmine, the present herbal dyes (alizarin and henna) are cheap and safe and could be potential alternatives for staining F. hepatica and other trematodes.