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Biomedical subjects

Monika Raulf-Heimsoth

Publications and source records attributed to Monika Raulf-Heimsoth.

At least 19 recordsLinked to original sources

Sensitization due to gum arabic (Acacia senegal): the cause of occupational allergic asthma or crossreaction to carbohydrates?

BACKGROUND: A pharmaceutical industry worker was exposed to dust of gum arabic in the tablet coating plant and complained of work-related shortness of breath, chest tightness, runny nose, itching and redness of the eyes. This case was investigated for allergy to gum arabic and compared with a control group. The aim of the study was to identify the IgE-binding components responsible for the work-related symptoms. METHODS: Skin prick tests (SPTs)and specific IgE (sIgE) measurements with environmental and occupational allergens, spirometry and a specific bronchial challenge with gum arabic were performed. One hundred and nineteen control subjects underwent SPT with gum arabic and 43 controls were tested for sIgE. Crossreactivity between gum arabic and horse radish peroxidase was investigated by IgE CAP inhibition. A combined procedure of immunoblotting and periodate treatment was applied to identify the epitope nature of gum arabic. RESULTS: Allergy to gum arabic was shown by SPT, presence of sIgE and a positive bronchial challenge with gum arabic. Sensitization to gum arabic was demonstrated by SPT or sIgE in 7 and 5 controls, respectively. The results of inhibition with horse radish peroxidase, immunoblotting and periodate treatment suggest that gum arabic sIgE of the patient and 1 SPT-positive control subject were directed to the polypeptide chains of gum arabic. In contrast, gum arabic sIgE of the other controls reacted to carbohydrate components. CONCLUSIONS: Sensitization to gum arabic carbohydrate structures occurs casually in atopic patients with pollen sensitization without obvious exposure to gum arabic. This study suggests that allergy to gum arabic is mediated preferentially by IgE antibodies directed to polypeptide chains of gum arabic.

Adult↗

Irritative effects of fumes and aerosols of bitumen on the airways: results of a cross-shift study.

Possible health hazards of fumes and aerosols of bitumen are in discussion, and data on their adverse effects on human airways under current exposure conditions are limited. To assess the irritative effects of exposure to fumes and aerosols of bitumen on the airways, a cross-sectional cross-shift study was conducted including external and internal exposure measurements, spirometry and especially non-invasive methods like nasal lavage collection and induction of sputum in order to identify and evaluate more precisely inflammatory process in the upper and lower airways. The cross-shift study comprised 74 mastic asphalt workers who were exposed to fumes and aerosols of bitumen and 49 construction workers without this exposure as reference group. Questionnaire, spirometry, ambient monitoring and urinary analysis were performed. Humoral and cellular parameters were measured in nasal lavage fluid (NALF) and induced sputum. For data analysis, a mixed linear model was performed on the different outcomes with exposure group, time of measurement (pre-, post-shift), current smoking, German nationality and age as fixed factors and subjects as random factor. Based on personal exposure measurements during shift, mastic asphalt workers were classified into a low (< or =10 mg/m(3); n = 46) and a high (>10 mg/m(3); n = 28) exposure group. High exposure was accompanied by significant higher urinary post-shift concentrations of 1-hydroxypyrene and the sum of hydroxyphenanthrenes. Acute respiratory symptoms were reported more frequently in the high exposure group after shift. Significant cross-shift declines in lung function parameters (forced expiratory volume in 1 s [FEV(1) (% predicted)] and forced vital capacity [FVC (% predicted)]) were measured in mastic asphalt workers. Pre-shift FEV(1) (% predicted) and FVC (% predicted) were higher in the low exposure group. In pre- and post-shift NALF samples, interleukin (IL)-1beta-, IL-8- and total protein concentrations were lower in the low exposure group compared to the reference and the high exposure group. Pre- and post-shift neutrophil percentages in both nasal and sputum samples were also lower in the low exposure group. Significantly higher pre- and post-shift sputum concentrations of IL-8, IL-6, nitrogen oxide (NO) derivatives and total protein were detected especially in highly exposed workers. Irritative effects of exposure to fumes and aerosols of bitumen on the upper and lower airways were apparent, especially in mastic asphalt workers with exposure above 10 mg/m(3).

Adult↗

Occupational endotoxin-exposure and possible health effects on humans.

BACKGROUND: Endotoxins are commonly found at workplaces where large amounts of bioaerosols are generated. In Germany, especially since the Ordinance on Safety and Health Protection related to work involving biological agents (Biostoff-Verordnung) became effective (1999), threshold limit values are widely discussed. Up to the present, endotoxin values are measured with non-uniform methods and therefore values are of limited benefit for classification of exposure groups. In Germany there is no threshold limit value for endotoxin. METHODS: Relevant literature of the last 20 years was selected from Medline and discussed. RESULTS: In this review we focused on the impact of endotoxin exposure on human health with special respect to the measurements on workplace and methodological aspects of endotoxin determination. Methods for sampling and endotoxin determination have to be validated, optimized, and standardized first. CONCLUSION: The adverse health effects of endotoxins are known, standardization of measurements is a necessary goal and protection measures should be established immediately.

Agriculture↗

Bites of the European pigeon tick (Argas reflexus): Risk of IgE-mediated sensitizations and anaphylactic reactions.

BACKGROUND: Local and systemic reactions can occur after bites of Argas reflexus (Argas), a soft tick parasitizing pigeons. OBJECTIVE: Risk assessment of IgE-mediated sensitizations and systemic reactions after Argas bites. METHODS: Case histories, skin prick tests (SPTs) with a whole-body extract of Argas containing major allergen Arg r 1, and common inhalants and specific IgE measurements were obtained from 148 subjects who had had Argas bites and 20 volunteers as a control group. RESULTS: Systemic reactions (urticaria, angioedema, dyspnea, cardiovascular dysregulation, unconsciousness) were reported in 12 of 148 (8%); 146 of 148 (99%) had local reactions. Atopy was found in 37 of 146 (25%) with local reactions and 3 of 12 (25%) with systemic reactions. SPT to Argas was positive in 24 of 148 (16%) with a high proportion of atopics 10 of 24 (42%); specific IgE to Argas was detectable in 12 of 135 (8% of 148) with moderate concordance to systemic reactions. No positive SPT or specific IgE results to Argas were obtained in the control group. Immunoblotting of 23 sera revealed an IgE-binding protein in 19 of 23 sera (82%) at 22 kd, indicating a major allergen of Argas. CONCLUSION: Severe anaphylactic reactions were infrequently (approximately 8%) found after bites of the soft tick Argas reflexus. Atopy is a risk factor for skin sensitizations to Argas, but not for systemic reactions after bites by Argas. Using a whole-body extract of Argas, diagnosis through SPT and specific IgE is hampered by false-negative and irrelevant positive results, particularly in atopy.

Adolescent↗

Relevance of the recombinant lipid transfer protein of Hevea brasiliensis: IgE-binding reactivity in fruit-allergic adults.

BACKGROUND: Lipid transfer proteins (LTPs) are relevant allergens in certain plants. The role of the LTP of Hevea brasiliensis in the latex-fruit syndrome is widely unknown. OBJECTIVE: To study IgE reactivity with recombinant Hevea LTP in sera of fruit-allergic adults with and without natural rubber latex (NRL) allergy. METHODS: An LTP-specific complementary DNA of H brasiliensis leaves was amplified, subcloned into the pMAL expression system, and analyzed. The recombinant protein was coupled to ImmunoCAP, and the IgE-binding properties were studied in sera of 10 NRL-allergic patients without symptoms to fruit and 48 atopic patients with fruit allergy. Eleven of these 48 patients were also allergic to NRL, 14 displayed sensitization to NRL without symptoms on NRL exposure so far, and 23 had neither symptoms nor IgE antibodies to NRL. RESULTS: After expression in Escherichia coli, a soluble maltose-binding protein-rHev b 12 fusion protein was isolated and coupled to ImmunoCAP to determine rHev b 12 specific IgE reactivity. rHev b 12 specific IgE binding was found in 3 fruit-allergic patients with NRL sensitization (0.68, 0.88, and 0.96 kU/L) and in 3 fruit-allergic patients without NRL sensitization (1.58, 2.25, and 2.27 kU/L). The remaining 52 serum samples and all maltose-binding protein control test results were negative (< 0.35 kU/L). CONCLUSIONS: In these patients, rHev b 12 specific IgE reactivity seems to result from common cross-reactive epitopes with some of the fruit LTPs tested and underscores only an involvement in co-recognition. No clinical relevance of IgE binding to the LTP of H brasiliensis in association with NRL allergy was detected.

Adolescent↗

Patterns of latex allergen recognition in children sensitized to natural rubber latex.

Single recombinant latex allergens permit the study of the pattern of sensitization to individual allergens. We aimed to quantify the IgE-response to individual latex allergens in children sensitized to latex. The study group included 31 latex-sensitized children: 26 operated at least twice, 20 of them with spina bifida; two children with one operation and three atopic non-operated children. IgE antibodies to rHev b 1, rHev b 3, rHev b 5, rHev b 6.01, rHev b 7.02 and rHev b 8, coupled to ImmunoCAPs, were measured in each serum. IgE responses to rHev b 1, rHev b 5 and rHev b 6.01 were found in 17 children each, and their mean +/- s.d. levels were 5 +/- 7.4, 16.8 +/- 14 and 10 +/- 18 kU/l, respectively. IgE responses to rHev b 3 (4 +/- 5.4 kU/l) were found in eight children. Two children had IgE to rHev b 7 (1.7 and 3.2 kU/l), and none to rHev b 8. Four sera were negative to all tested recombinant allergens. We divided the patients in three groups: sensitized only to rHev b 1, sensitized only to rHev b 5 and/or rHev b 6.01, and sensitized to both rHev b 1 and to rHev b 5 and/or rHev b 6.01. The three groups had the same profile of clinical features. Hev b 5 induces the quantitatively higher IgE responses in children with multiple surgeries sensitized to latex. Responses to Hev b 6.01 equal those of Hev b 1.

Adolescent↗

Basophil Activation Test and specific IgE measurements using a panel of recombinant natural rubber latex allergens to determine the latex allergen sensitization profile in children.

There are no documented studies that describe natural rubber latex (NRL) sensitization in children with a history of surgical intervention but without any congenital malformation (urogenital anomalies, spina bifida, etc.), although some authors have studied NRL allergy in children without a history of surgical intervention. The aim of this work was to evaluate the sensitization profile to single NRL allergens in children without spina bifida and without repeated surgical interventions, by using different recombinant and natural latex allergens in two analytical techniques: specific serum immunoglobulin E (IgE) quantification and flow cytometry determination of activated basophils expressing CD63, after stimulating cells from patients with NRL allergens. A total of 23 patients and 10 healthy children were selected. Conjunctival and in-use NRL provocation tests were carried out, as well as specific IgE determination in all patients' and controls' sera with the recombinant NRL allergens: rHev b 1, rHev b 2, rHev b 3, rHev b 5, rHev b 6.01, rHev b 6.02, rHev b 8, rHev b 9 and rHev b 11 and with NRL (k82) using appropriate ImmunoCAPs. The Basophil Activation Test (BAT) was performed with whole latex extract and with the recombinant allergens rHev b 5 and rHev b 6.01, as well as with the natural allergen Hev b 6.02. The sensitivity and the specificity of NRL-specific IgE (k82) were 100%. Positive IgE responses to rHev b 5 were found in sera of 10 children, to rHev b 6.01 in 16 and for rHev b 6.02 in 15 children's sera. Specific IgE to rHev b 8 was found in four sera of the children. We only found significant differences in sensitization to rHev b 5 in children with two or more surgical interventions compared with the non-intervened group or those with only one intervention. Specific IgE in sera of children with latex-fruit syndrome recognized rHev b 6.02, but not to rHev b 11. The patients sensitized to Hev b 8, Hev b 9 and/or Hev b 11 were atopic. The four patients presenting a positive response to the NRL profilin Hev b 8 were allergic to pollen. The BAT against whole NRL extract was positive in 22 of 23 children; against rHev b 5 in 14 of the patients studied; against rHev b 6.01 in seven cases and against nHev b 6.02 in 19 children. In all the control subjects, the results using this technique were negative. If combined rHev b 5, rHev b 6.01 and nHev b 6.02 together, BAT could detect 20 of the 23 children with latex allergy. The combined use of ImmunoCAP with all the recombinant NRL allergens and BAT with rHev b 5, rHev b 6.01 and nHev b 6.02, enabled the identification of NRL allergy in 22 of 23 patients. There is a positive and significant correlation between sensitization to Hev b 5 and the number of interventions. BAT and allergen-specific IgE determination could be used as first-line in vitro diagnostic tests in patients with NRL allergy.

Adolescent↗

Assessment of DNA damage in WBCs of workers occupationally exposed to fumes and aerosols of bitumen.

We conducted a cross-shift study with 66 bitumen-exposed mastic asphalt workers and 49 construction workers without exposure to bitumen. Exposure was assessed using personal monitoring of airborne bitumen exposure, urinary 1-hydroxypyrene (1-OHP), and the sum of 1-, 2 + 9-,3-,4-hydroxyphenanthrene (OHPH). Genotoxic effects in WBC were determined with nonspecific DNA adduct levels of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) and the formation of DNA strand breaks and alkali-labile sites. Concentration of fumes and aerosols of bitumen correlated significantly with the concentrations of 1-OHP and OHPH after shift (r(s) = 0.27; P = 0.03 and r(s) = 0.55; P < 0.0001, respectively). Bitumen-exposed workers had more DNA strand breaks than the reference group (P < 0.0001) at both time points and a significant correlation with 1-OHP and OHPH in the postshift urines (r(s) = 0.32; P = 0.001 and r(s) = 0.27; P = 0.004, respectively). Paradoxically, we measured higher levels of DNA strand breaks, although not significant, in both study groups before shift. 8-OxodGuo adduct levels did not correlate with DNA strand breaks. Further, 8-oxodGuo levels were associated neither with personal exposure to bitumen nor with urinary metabolite concentrations. Significantly more DNA adducts were observed after shift not only in bitumen-exposed workers but also in the reference group. Only low-exposed workers had significantly elevated 8-oxodGuo adduct levels before as well as after shift (P = 0.0002 and P = 0.02, respectively). Our results show that exposure to fumes and aerosols of bitumen may contribute to an increased DNA damage assessed with strand breaks.

Adult↗

Cellular uptake and cytotoxic potential of respirable bentonite particles with different quartz contents and chemical modifications in human lung fibroblasts.

Considering the biological reactivity of pure quartz in lung cells, there is a strong interest to clarify the cellular effects of respirable siliceous dusts, like bentonites. In the present study, we investigated the cellular uptake and the cytotoxic potential of bentonite particles (Ø< 10 microm) with an alpha-quartz content of up to 6% and different chemical modifications (activation: alkaline, acidic, organic) in human lung fibroblasts (IMR90). Additionally, the ability of the particles to induce apoptosis in IMR90-cells and the hemolytic activity was tested. All bentonite samples were tested for endotoxins with the in vitro-Pyrogen test and were found to be negative. Cellular uptake of particles by IMR90-cells was studied by transmission electron microscopy (TEM). Cytotoxicity was analyzed in IMR90-cells by determination of viable cells using flow cytometry and by measuring of the cell respiratory activity. Induced apoptotic cells were detected by AnnexinV/Propidiumiodide-staining and gel electrophoresis. Our results demonstrate that activated bentonite particles are better taken up by IMR90-cells than untreated (native) bentonite particles. Also, activated bentonite particles with a quartz content of 5-6% were more cytotoxic than untreated bentonites or bentonites with a quartz content lower than 4%. The bentonite samples induced necrotic as well as apoptotic cell death. In general, bentonites showed a high membrane-damaging potential shown as hemolytic activity in human erythrocytes. We conclude that cellular effects of bentonite particles in human lung cells are enhanced after chemical treatment of the particles. The cytotoxic potential of the different bentonites is primarily characterized by a strong lysis of the cell membrane.

Air Pollutants↗

Rapid detection of the SPINK5 polymorphism Glu420Lys by real-time PCR technology.

BACKGROUND: The SPINK5 gene, encoding the serine protease inhibitor LEKTI maps to chromosome 5q32, and has been suggested to be a locus predisposing to atopic diseases in general. The Glu420Lys variant showed significant association with atopy, asthma and atopic dermatitis in recent studies. AIMS OF THE STUDY: Development of a high throughput assay to analyse the polymorphism G1258A (Glu420Lys) in exon 14 of the SPINK5 gene followed by the validation using samples of 235 latex-allergic health care workers (HCWs) with (N=63) and without asthma (N=172), and 80 non-atopic controls. METHODS: Twenty DNA samples were first analysed by a polymerase chain restriction fragment analysis (RFLP) using Hph I to generate defined control DNAs which were used for the development of the assay suitable for the detection of the Glu420Lys variant by LightCycler technology. RESULTS AND CONCLUSIONS: 315 samples were successfully screened with this new assay. The temperatures in the melting analysis of the SPINK5 exon 14 PCR product were characteristic to the probes hybridised to the mutant (AA) at 51.5 degrees C and to the wild-type (GG) at 59.5 degrees C. The fast and reliable mutation detection in the tested samples makes this high-speed method suitable for larger epidemiological studies.

Adult↗

Natural rubber latex allergy: antigen specific IgE in Polish blood donors, prevalence and risk factors--preliminary data.

OBJECTIVES: There is insufficient data on the prevalence of latex allergy in the general population in Poland. The aim of the study was to evaluate natural rubber latex (NRL) sensitization and its risk factors among Polish blood donors. MATERIALS AND METHODS: The study group comprised 1000 consecutive blood donors attending the Regional Centre for Transfusion Medicine. Total IgE and allergen specific IgE to NRL (asNRL-IgE) were assayed using the EIA (enzyme immunoassay) test. In the subjects with asNRL-IgE equal to or above Class II further surveys were performed: questionnaire, skin prick tests (SPT) to common allergens, NRL and food allergens cross reactive to NRL, asNRL-IgE with FDA (Food and Drug Administration) approved FEIA (fluorescent enzyme immunoassay) test and antigen profile of asNRL-IgE. RESULTS: asNRL-IgE (EIA) was observed in 17.9% of blood donors. Only 10% of positive results equal to or above Class II were confirmed with the FEIA test. The positive FEIA results were confirmed with SPT to NRL in 60% and the negative FEIA results in 5.4% of cases. The specific IgE to Hev b 3, 5, 6 and 13 were observed in the subjects with occupational exposure to NRL. The highest concentration of Hev b 8 was observed in the subjects not symptomatic to NRL with positive SPT to grass and tree pollens. CONCLUSION: The EIA test showed the high prevalence of antigen specific IgE to NRL among Polish blood donors. There was low concordance between EIA and FEIA tests. Therefore, the EIA test should be used only for screening purposes along with the establishment of sensitivity and specificity of the method. Elevated total IgE level, active tobacco smoking, and history of atopic disease symptoms were revealed to be risk factors for the presence of asNRL-IgE.

Adult↗

A new method to bind allergens for the measurement of specific IgE antibodies.

BACKGROUND: Detection of allergen-specific IgE antibodies in patients' sera plays a key role for the diagnosis of IgE-mediated allergy. If no validated test system is available, diagnostic tools must be developed, usually by coupling or binding the allergens to a solid phase. Streptavidin ImmunoCAP is a new solid phase for binding of allergens which can be used in the Pharmacia CAP system. OBJECTIVE: It was the aim of this study to assess the diagnostic validity of Streptavidin ImmunoCAP. METHODS: Biotinylation and allergen concentration for binding to Streptavidin ImmunoCAP were optimized and IgE obtained with natural rubber latex, obeche wood, wheat and rye flour Streptavidin ImmunoCAP were compared with the results of ImmunoCAP and Enzyme Allergo-Sorbent Test (EAST) using sera from patients complaining of workplace-related respiratory symptoms. RESULTS: While the relation of biotin-label and protein was critical (best results were obtained with a 5- fold molar excess), labelled protein for coupling to streptavidin ImmunoCAP was applicable in a wide concentration range. On average, IgE values with streptavidin ImmunoCAP were as high as with ImmunoCAP but considerably higher than values obtained by EAST. CONCLUSION: Streptavidin ImmunoCAP is a valuable tool for sensitive and specific measurement of IgE binding to new allergens superior to cellulose disk-based methods.

Allergens↗

Hev b 5 and Hev b 13 as allergen markers to estimate the allergenic potency of latex gloves.

BACKGROUND: Sensitization to natural rubber latex has been linked to proteins from medical latex gloves. Various assays to estimate the amount of residual allergenic proteins extractable from latex gloves to assess their potential exposure hazard have inherent weaknesses. OBJECTIVE: This investigation was aimed at developing 2-site immunoenzymetric assays and identifying appropriate protein markers to assess the allergenic potential of latex gloves. METHODS: The presence of 6 latex allergens--Hev b 1, 2, 3, 5, 6, and 13--was measured in a cross-section of commercial latex medical gloves by using monoclonal and polyclonal antibody-based 2-site immunoenzymetric assays. The overall allergenic potential of these gloves was assessed by IgE-inhibition assay. Stepwise multiple regression analyses were performed to identify marker allergens that best explained the variation in latex glove allergenicity. RESULTS: All 6 latex allergens were detected in at least some of the glove samples. Hev b 5 and Hev b 13 were identified as the marker allergens that combined best to explain the variation in the glove allergenicity. The significant multiple correlation (R=0.855) between these 2 markers and glove allergenic potency forms the basis of an assay to gauge latex glove allergenicity. CONCLUSION: The overall allergenic potential of latex gloves can be estimated by using Hev b 5 and Hev b 13 as indicator allergens. The correlation between glove allergenicity and the level of these allergens was maintained for low-protein gloves (<200 microg/g). This estimation of glove allergenicity was superior to that obtained by using total protein readings.

Allergens↗

Evaluation of patatin as a major cross-reactive allergen in latex-induced potato allergy.

BACKGROUND: Potential cross-reactions between natural rubber latex and fruit/vegetable specific immunoglobulin (Ig)E antibodies have been reported for many years. This study was designed to investigate the molecular basis of acquired food sensitization focusing on the storage protein patatin and the patatin-like latex protein Hev b 7. OBJECTIVE: The amount of potato-specific IgE in the serum of latex-allergic health care workers and children with atopic dermatitis was determined to evaluate cross-reactivity between Hev b 7 and patatin. Additionally, the stability of potato patatin to digestion was investigated. METHODS: Human serum was tested on its reactivity to latex and potato proteins by IgE immunoblotting after one-dimensional (1-D) and 2-D electrophoresis. Latex- and potato-specific IgE concentrations were measured in fluorescence enzyme immunoassays (CAP, Pharmacia, Uppsala, Sweden). Further, potato patatin was chromatographically isolated to perform auto-inhibition tests. Stability of patatin to degradation was determined by digestion in vitro. RESULTS: Patatin was identified as major cross-reactive potato allergen by N-terminal sequencing. Seventy-five percent of the potato-sensitized people reacted with patatin in 1-D immunoblots, and 25% of the positive reactions to Hev b 7 could be blocked by preincubation of the patients' sera with purified potato patatin. Examination of children with atopic dermatitis showed that most sera contained patatin-specific IgE, whereas no Hev b 7-specific IgE was detected. Finally, patatin has been found partially stable to digestion in vitro. CONCLUSIONS: Patatin was identified as a major cross-reactive protein in latex-associated potato allergy and appears to be relevant for atopic dermatitis. Therefore, patatin could be a suitable marker for the determination of potato sensitization, and it may also constitute an important food allergen. Cross-reactivity between Hev b 7 and patatin was restricted to primarily latex-sensitized adults, suggesting a different mechanism of sensitization in children with atopic dermatitis.

Adolescent↗