Adam B. Czuppon

The rubber elongation factor of rubber trees (Hevea brasiliensis) is the major allergen in latex.

BACKGROUND Allergy to latex-containing articles is becoming more and more important because it can result in unexpected life-threatening anaphylactic reactions in sensitized individuals. METHODS A protein of 58 kd with an isoelectric point of 8.45 was purified from raw latex and from latex gloves and identified as the major allergen, completely blocking specific IgE antibodies in the serum of latex-sensitized subjects. The allergen is a noncovalent homotetramer molecule, in which the 14.6 kd monomer was identified, by amino acid composition and sequence homologies of tryptic peptides, to be the rubber elongation factor found in natural latex of the Malaysian rubber tree. RESULTS Competitive immunoinhibition tests showed that the starch powder covering the finished gloves is the airborne carrier of the allergen, resulting in bronchial asthma on inhalation. The purified allergen can induce allergic reactions in the nanogram range. CONCLUSION The identification of the allergen (Hev b I) may help to eliminate it during the production of latex-based articles in the future.

Respiratory and other hazards of isocyanates.

Isocyanates are increasingly being used for manufacturing polyurethane foam, elastomers, adhesives, paints, coatings, insecticides, and many other products. At present, they are regarded as one of the main causes of occupational asthma. The large number of workers who are exposed to these chemicals have a concentration-dependent risk of developing chronic airway disorders, especially bronchial asthma. Different pathophysiologic mechanisms are involved. Immunoglobulin E (IgE)-mediated sensitization and irritative effects have been clearly demonstrated in both exposed subjects and animals. Presumably, neural inflammation due to neuropeptide release of capsaicin-sensitive afferent nerves is crucial. We collected data on 1780 isocyanate workers who had been examined by our groups. Of them 1095 (including subjects from outpatient departments) had work-related symptoms, predominantly of the respiratory tract. Specific IgE antibodies were found in 14% of the 1095 subjects. The methacholine challenge test was shown to be an inadequate predictor of the results of inhalative isocyanate provocation tests in workers and in asthmatic controls. Isocyanate (toluene diisocyanate TDI) air concentrations of 10 ppb (0.07 mg/m3) and 20 ppb (0.14 mg/m3), respectively, did not cause significant bronchial obstruction in the majority of previously unexposed asthmatics with bronchial hyperreactivity. IgG-mediated allergic alveolitis, a rare disease among isocyanate workers, was found in approximately 1 % of the symptomatic subjects. Experimental studies exhibit dose-dependent toxic effects and give evidence for tachykinin-mediated bronchial hyperreactivity after exposure to isocyanates. The clinical role of genotoxic effects of isocyanates and their by-products demonstrated here in vitro and in vivo has yet to be clarified.

Health risk in hospitals through airborne allergens for patients presensitised to latex

Two nurses had to stop practicing because of severe work-related allergic respiratory or systemic symptoms. Both developed similar symptoms within a short time when they attended hospitals as outpatients or visitors or consulted a doctor or dentist, even when they had no direct contact with latex articles. The airborne latex allergen concentration in one room that induced symptoms was 128 ng per m3. In skinprick tests both women showed positive reactions to the phosphated cornstarch powder used to prevent latex gloves sticking together, but only when the powder had been in contact with latex. This powder seems a likely vehicle for the airborne latex allergen.

Characterization of soybean allergens causing sensitization of occupationally exposed bakers

Fourteen bakers suffering from workplace‐related respiratory symptoms and sensitized to soybean were studied. Twelve of them were also allergic to wheat flour, 10 to rye flour, and five to α‐amylase of Aspergillus oryzae (Asp o 2). IgE estimation by RAST strongly indicated that the trypsin inhibitor and lipoxidase are major allergens of soybean. Various allergenic components could be characterized by immunoblotting after two‐dimensional electrophoresis. Our RAST and immunoblotting results show an interindividually different allergic response to inhalative soybean constituents, and that the trypsin inhibitor (20 kDa, pI˜4.5) is an important inhalative soybean allergen recognized by IgE antibodies in the sera of 86% of the examined sensitized bakers.

Diagnostic validation of specific IgE antibody concentrations, skin prick testing, and challenge tests in chemical workers with symptoms of sensitivity to different anhydrides.

BACKGROUND The diagnostic possibilities of sensitization to various acid anhydrides are limited because of the lack of standardized allergens for the different test systems. This makes the diagnosis of IgE-mediated sensitization caused by occupational exposure difficult. METHODS We prepared conjugates of human serum albumin with phthalic, maleic, trimellitic, and pyromellitic anhydrides to be used for IgE estimation by enzyme-allergosorbent test, skin prick tests, and nasal and bronchial challenge tests. Nine anhydride workers, who complained of various respiratory symptoms, were studied. RESULTS Of the nine workers, four had immediate-type skin test responses to one or more conjugates. All four subjects had elevated IgE concentrations in addition to two other workers. Three of six nasal challenges and four of nine bronchial challenges resulted in positive responses. All but one of the positive nasal or bronchial test responses were associated with elevated IgE levels. The seven positive challenge test results included five positive skin test responses. On the other hand, in all but two of the subjects with negative challenge test results, no specific IgE could be detected. In these two subjects the negative results were associated with low levels of IgE, and in one, with the absence of asthma. None of the results of tests with unconjugated anhydrides were positive. CONCLUSIONS Anhydrides investigated in this study can induce IgE-mediated hypersensitivity, which can be diagnosed by using the respective human serum albumin in estimation of specific IgE and in skin, nasal, and bronchial challenge tests. Estimation of IgE was demonstrated to be more sensitive than skin prick testing.

Increased Incidence of DNA Double-strand Breaks and Anti-ds DNA Antibodies in Blood of Workers Occupationally Exposed to Asbestos

Asbestos, proven to be carcinogenic in humans and animals, is reported to have no genotoxic effect. Asbestos workers have an increased risk of lung cancer, mesothelioma, and other tumours. Earlier findings showed that crocidolite can induce DNA strand breaks in cultured rat embryo cells as assessed by nick translation. We investigated DNA double-strand breaks in white blood cells (WBC) of ten workers occupationally exposed to asbestos. According to our results, obtained with neutral filter elution, individuals who had been exposed to asbestos fibres showed two to four times more DNA double-strand breaks (dsb) in white blood cells than ten non-exposed persons. The induced DNA fragments are of about 250 kb (compared to chromosomal DNA of Saccharomyces cerevisiae standard marker). Using additionally the chromosomal DNA protective method of agarose-plugs, DNA fragments in the range of 200 to 1000 kb have been found in the white blood cells of the same ten workers occupationally exposed to asbestos. In the white blood cells of non-exposed subjects no DNA fragments could be detected with this method. Compared to 51 non-exposed persons, elevated anti-ds DNA antibody concentrations were found in ten workers occupationally exposed to asbestos. The fact that workers occupationally exposed to asbestos have distinctly more double-strand breaks and anti-ds DNA antibodies could mean that an increased incidence of DNA-fragments may be an important indicator in the chronic effect of asbestos-associated carcinogenesis. Apparently, the chronic effects of asbestos observed here do not seem to be identical with that of previously reported acute in vitro effects.

A clinical and immunological study on 92 workers occupationally exposed to anhydrides

This study aimed at investigating sensitizing and hazardous effects of a new acid anhydride, pyromellitic dianhydride (PMDA), in addition to those of phthalic anhydride, maleic anhydride and trimellitic anhydride, in a group of 92 exposed workers in two German chemical plants. Of the 92 workers, 56 reported work-related complaints with a predominance of phlegm and dyspnoea in those exposed to anhydride dust for less than 1 year. Haemorrhagic rhinitis occurred only after a prolonged exposure of more than 15 years. Specific IgE antibodies to anhydride-HSA conjugates could be detected in 15 exposed subjects, 12 of whom had work-related symptoms. The IgE-positive group had significantly more impaired lung function parameters than the IgE-negative group. The proportion of IgE-positive subjects was highest in the groups with dyspnoea (5/18), cough (6/24) and rhinitis (11/44) whereas only 1 of 11 workers with haemorrhagic rhinitis had such antibodies. A follow-up study of 23 affected workers was performed after 10 months to assess clinical symptoms, lung function and IgE antibody levels. This follow-up study showed the absence of obstructive ventilation patterns in three out of six subjects in addition to cessation of symptoms in most initially affected workers who were no longer exposed. On the other hand, 14 workers under continuous exposure had comparable pathological findings on re-examination. Our results confirm that anhydrides including the lesser known PMDA, behave as respiratory irritants and as immediate-type sensitizers. They predominantly induced reversible symptoms in workers whose exposure stopped after a working period of about 0.7 years. Abnormal lung function parameters normalized in nearly 50% of these subjects.

DNA damage in human white blood cells after inhalative exposure to methylenediphenyl diisocyanate (MDI) — case report

A workplace-related inhalative exposure test in a challenge chamber was performed on an industrial worker in a methylenediphenyl diisocyanate (MDI) atmosphere from 5 to 20 ppb and the isolated white blood cell DNA was analysed by electrophoresis, anion-exchange chromatography and melting behaviour. The results of electrophoresis indicate that inhaled MDI induces double-strand breaks of DNA. Some of the DNA fragments were estimated to be in the region of 100-500 bp. Anion-exchange chromatography confirmed this finding. Following denaturation and rapid renaturation the results demonstrated that some DNA fragments are cross-linked by MDI. Comparing the melting curves before and after inhalative exposure in the challenge test chamber, genomic DNA revealed differences in the shape of the melting curve (hyperchromic effect). The results suggest that occupational MDI exposure could be associated with white blood cell DNA damage.

INCREASED INCIDENCE OF DNA DOUBLE-STRAND BREAKS IN LUNG AND LIVER OF RATS AFTER EXPOSURE TO CROCIDOLITE ASBESTOS FIBERS

AbstractThe aim of this study was to find out whether UICC crocidolite asbestos fibers induce in vivo DNA double-strand breaks in the nuclei of cells in lung and liver of rats. The effect of crocidolite on chromosomal DNA of rats exposed to crocidolite fibers was analyzed by pulsed-field gel electrophoresis (PFGE) using the chromosomal DNA-protective agarose-plugs method and laser densitometric evaluation of stained DNA bands. The profiles of laser densitometric scanning showing PFGE separation of DNA for exposed rats are right-shifted in liver and lung compared with the controls of the same age. DNA from the liver of nonexposed rats showed one peak in the range of 250–850 kb. Compared to this finding, at 4, 13, and 16 mo after crocidolite instillation DNA of rats showed shorter DNA fragments in the range of 250–700 kb. This indicates a more intact DNA in controls. At 4, 13, and 16 mo after crocidolite instillation, we also found DNA fragments in exposed lungs in the region between 250 and 2200 kb produce...

Allergic reaction after eating α‐amylase (Asp o 2)‐containing bread A case report

A 29‐year‐old female bakery shop assistant was occupationally sensitized to flour allergens and Aspergillusα‐amylase (Asp o 2). The latter represents a strongly allergenic component of routinely used baking additives. The patient had repeatedly responded to the consumption of white bread with rhinitis, conjunctivitis, and, occasionally, wheal and flare reactions. She underwent allergologic investigations including oral challenge tests with commercially available bread loaves. Elevated specific IgE antibodies against bread extracts, Asp o 2, and flour allergens were detectable in her serum. The provocation test with bread resulted in a running nose together with a strong increase in nasal resistance. All symptoms subsided about 3 h after the challenge. None of the above symptoms could be observed when bread free of Aspergillusα‐amylase was administered. This outcome provides evidence of a clinically relevant persistent allergenicity to Asp o 2 in bread.