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Biomedical subjects

R D Tee

Publications and source records attributed to R D Tee.

At least 19 recordsLinked to original sources

Allergen and dust exposure as determinants of work-related symptoms and sensitization in a cohort of flour-exposed workers; a case-control analysis.

OBJECTIVES: To estimate the incidence of specific IgE sensitization and allergic respiratory symptoms among UK bakery and flour mill workers; and to examine the roles of flour aeroallergen and total dust exposures in determining these outcomes. METHODS: A cohort of 300 new employees, without previous occupational exposure to flour, were followed prospectively for a median (range) of 40 (1-91) months. Cases-defined as those developing work-related symptoms or a positive skin prick test to flour or alpha-amylase during follow up--were compared with controls, matched for duration of employment. Exposures to flour aeroallergen and total inhalable dust were estimated using a questionnaire and personal sampling techniques. RESULTS: Incidence rates for work-related eye/nose and chest symptoms were 11.8 and 4.1 cases per 100 person years (py), respectively. Fewer employees developed positive skin prick tests to flour (2.2 cases per 100 py) or alpha-amylase (2.5 cases per 100 py). Positive skin tests to occupational allergens were more common among those with new work-related symptoms. There were clear relationships between the risks of developing work-related symptoms or a positive skin prick test and three categories of estimated exposure to total dust or flour aeroallergen. Atopic employees were more likely to develop a positive skin prick test-but not work-related symptoms. These findings were unaffected by age, sex or cigarette smoking. CONCLUSIONS: In this population, many work-related symptoms which develop after first employment in modern UK bakeries or flour mills were not accompanied by evidence of IgE sensitization to flour or alpha-amylase. Although average dust exposures were within current occupational standards, the risks of development of upper and lower respiratory symptoms and of specific sensitization were clearly related to total dust and/or flour aeroallergen exposure. The incidence of work-related chest symptoms in the presence of a positive skin test to flour or alpha-amylase in this setting was approximately 1 case per 100 py.

Adult↗

Analysis of allergens in rat fur and saliva.

BACKGROUND: The fur is an important source of allergens in many mammal species, but this source has not been extensively studied in rats. Rat room dust contains high-molecular-weight allergens that have been found to cross-react with fur and its presumed salivary contaminants. The role of rat fur and saliva as a source of respiratory allergens merits further investigation. The objective was to describe the allergens present in the fur and saliva of male rats. METHODS: Allergen extracts were prepared from the saliva and fur of the pelts from adult male rats. Immunoblotting was used to describe the allergens present in rat fur and saliva with serum from 76 and 25 individuals, respectively. RESULTS: There was considerable variation between the individuals in the binding of IgE to the separated fur and saliva allergens. Immunoblot analysis identified 23 allergens in rat fur. "Major" allergens were found at the "origin", and at 55, 51, 19, and 17 kDa, and "intermediate" allergens at 74, 67 (probably albumin), and 21.5 (diffuse) kDa. Seventeen salivary allergens were described with "major" allergens at 21.5, 19.5, 19, 18, and 17.5 kDa. Many subjects had IgE to the 67 kDa (56%) and 43 kDa (64%) allergens but the density of staining was weak. CONCLUSION: Rat fur and saliva are the sources of many allergens. Fur contained five "major" allergens and was a complex source of allergens of relatively high molecular weight (>22 kDa). The most important salivary allergens have molecular weights of less than 22 kDa. Fur is the most probable source of the high-molecular-weight allergens found in rat room dust.

Allergens↗

Allergen exposure, atopy and smoking as determinants of allergy to rats in a cohort of laboratory employees.

This study aimed to examine the relationship between exposure to rat urinary allergens, atopic status, smoking and the development of allergic symptoms and specific sensitization. It is a case-referent analysis of a cohort of 342 newly employed laboratory animal workers. Cases comprised persons developing symptoms of laboratory animal allergy or a positive skin prick test to rat urinary allergens; each was matched with up to two asymptomatic referents. Subjects were assigned to categories of exposure based on measurements of airborne rat urinary allergens. Of the cases, 80% reported that their symptoms started within 2 yrs of employment. The odds ratio (OR) for development of each symptom type (respiratory, eye or nose and skin) and of an immediate skin test reaction was increased in those with direct contact with rats. A gradient of increasing OR for the development of any such symptom across exposure categories was found; for respiratory symptoms and skin test reactions the OR for subjects in the highest exposure category were lower than those in intermediate categories, a pattern attenuated when the analysis was confined to outcomes developing within 2 yrs of first exposure. Atopy increased the OR of most outcomes as did cigarette smoking, although there was no evidence of a relationship between smoking and the development of a specific skin test reaction. In conclusion, allergen exposure was confirmed as the most important determinant of laboratory animal allergy; by implication, measures to reduce exposure may be the most effective means to reduce its incidence.

Adult↗

Specific IgE to isocyanates: a useful diagnostic role in occupational asthma.

BACKGROUND: Isocyanates are the most frequent cause of occupational asthma in industrialized countries. OBJECTIVE: We sought to investigate the utility of specific IgE measurement in the diagnosis of isocyanate-induced asthma. METHODS: Fifty-eight of 101 patients referred for investigation were diagnosed as having isocyanate-induced occupational asthma by means of history, serial peak flow records, and bronchial provocation tests. Specific IgE antibodies to toluene diisocyanate:human serum albumin (HSA), diphenylmethane diisocyanate:HSA, and hexamethylene diisocyanate: HSA were measured in all patients by Phadebas RAST. RESULTS: Twenty patients had a RAST ratio of 2 or greater to at least one isocyanate. Thirteen (28%) of the 46 patients with a positive provocation test response had a RAST ratio of 2 or greater, and nine (20%) had a RAST ratio of 3 or greater. Raising the RAST cut-off from 2 or greater to 3 or greater reduced its sensitivity but increased the specificity of the test to 100%. RAST measurement was most likely to be positive within 30 days of exposure. Serial measurements suggested that the half-life of the IgE antibodies was approximately 6 months. Evidence of cross-reactivity between isocyanate RAST responses was found in eight subjects. CONCLUSION: Specific IgE to isocyanates is a more specific than sensitive index of occupational asthma. With a RAST score of 3 or greater, it is wholly specific and therefore diagnostic of isocyanate-induced asthma. The sensitivity of specific IgE measurement is highest when blood is taken less than 30 days from last exposure, which is consistent with the observed half-life.

Adolescent↗

Measurement of exposure to mouse urinary proteins in an epidemiological study.

OBJECTIVES: To develop an assay to measure airborne mouse urinary protein (MUP) and to assess the occupational exposure to MUP in the workforce of three establishments as part of an epidemiological study examining the influence of aeroallergen exposure on the development of allergic respiratory disease. METHODS: Personal air samples were collected from nine exposure groups during a workshift. A sensitive and reproducible competitive inhibition assay, which used rabbit antisera specific for MUP, was developed and used to measure the occupational exposure to MUP. RESULTS: The personal measurements of MUP showed that people with direct contact with mice (animal technicians) had the highest exposure followed in decreasing order by those working with anaesthetised animals or their tissue (postmortem workers and scientists) and those with indirect contact with mice (supervisors, office workers, and slide production workers). The only difference in concentrations of MUP between the three establishments were found for cage cleaners, which reflected differences in working practises for this exposure category. Air samples collected during the performance of specific tasks showed that high exposures to MUP were associated with handling mice, indirect contact with mice, and washing floors. CONCLUSIONS: Exposure to mouse urinary proteins has been measured in the occupational environment. This information can be used to determine the relation between exposure to MUP and the development of allergic and respiratory disease.

Air Pollutants, Occupational↗

Thunderstorm-related asthma--the epidemic of 24/25 June 1994.

BACKGROUND: A large epidemic of asthma occurred following a thunderstorm in southern and central England on 24/25 June 1994. A collaborative study group was formed. OBJECTIVES: To describe the epidemic and the meteorological, aerobiological and other environmental characteristics associated with it. METHODS: Collation of data from the Meteorological Office, the Pollen Research Unit, the Department of the Environment's Automatic Urban Network, from health surveillance by the Department of Health and the National Poisons Unit, from clinical experience in general practice and hospitals, and from an immunological study of some of the affected cases from north east London. RESULTS: The thunderstorm was a Mesoscale Convective System, an unusual and large form of storm with several centres and severe wind gusts. It occurred shortly after the peak grass pollen concentration in the London area. A sudden and extensive epidemic occurred within about an hour affecting possibly several thousand patients. Emergency services were stretched but the epidemic did not last long. Cases had high serum levels of IgE antibody to mixed grass pollen. CONCLUSION: This study supports the view that patients with specific IgE to grass pollen are at risk of thunderstorm-related asthma. The details of the causal pathway from storm to asthma attack are not clear. Case-control and time series studies are being carried out.

Adult↗

Reduction of exposure to laboratory animal allergens in the workplace.

OBJECTIVES: Evidence is now accumulating that the prevalence of allergy to laboratory animals is related to the intensity of exposure to animal allergens. Whilst airborne animal allergen concentrations may be influenced by the litter type, cage design and stock density, the effectiveness of methods to reduce personal exposure has not been objectively assessed. METHODS: Air samples were collected at 2 L/min and 180 L/min onto polytetrafluoroethylene (PTFE) filters and the rat urinary aeroallergen (RUA) and mouse urinary aeroallergen (MUA) concentrations were measured by radioallergosorbent test (RAST) inhibition. RESULTS: When 545 mice (11.l mice/m3) were housed in ventilated cages (Thoren Maximiser cage system) operated at positive pressure to the environment, the static MUA concentration (n = 24, median = 0.10 microg/m3) was reduced sevenfold when compared with conventional cage systems (n = 12, median =0.67 microg/m3, P< 0.001). MUA could be further reduced if the ventilated cage system was operated at lower pressure; static samples (n = 1) collected at 180 L/min at negative, ambient and positive pressure registered < 0.003, 0.02 and 0.28 microg/m3, respectively. During cleaning out, the intensity of personal exposure to RUA was apparently reduced twofold when soiled litter was removed by vacuum (n = 17, median = 22.87 microg/m3) when compared with tipping (n = 18, median = 38.15 microg/m3), P = 0.002) although the task took twice as long to perform. The RUA exposure associated with handling rats was'reduced 25-fold when performed in a ventilated cabinet (n = 21, median = 2.67 microg/m3) compared with handling of rats on an open bench (n = 17, median = 54.39 microg/m3, P = 0.0001). CONCLUSIONS: Effective reduction of exposure to animal allergens can now be achieved by the use of ventilated systems both for housing and handling rats and mice providing safety equipment is used correctly. The vacuum removal of soiled litter during the task of cleaning out was less efficient and additional respiratory protection is therefore recommended for this procedure.

Air Pollutants, Occupational↗

Identification of the major water/salt insoluble wheat proteins involved in cereal hypersensitivity.

BACKGROUND: Several studies have investigated water/salt soluble proteins which comprise 50% of the proteins in wheat. The remaining 50% of wheat proteins, are water/salt insoluble proteins of which there is limited information on their role in cereal hypersensitivity. OBJECTIVES: To investigate the allergenicity of the water/salt insoluble gliadin and glutenin proteins (prolamins). METHODS: RAST, electrophoresis and Western blotting were used to identify water/salt insoluble wheat allergens. Competitive RAST inhibition was conducted to investigate cross-reactivity between prolamins and water/salt soluble wheat proteins. RESULTS: Specific IgE to alpha-gliadin and to total glutenins were detected in all sera. IgE to beta-, gamma-, fast omega-, and slow omega-gliadin were present in lower numbers of sera. Prolamin allergens of 90-11 kDa were identified by immunoblotting. Water/salt soluble proteins crossreacted with alpha-gliadin and total glutenins. CONCLUSIONS: Individuals who are hypersensitive to water/salt soluble wheat proteins produce specific IgE to water/salt insoluble wheat proteins. Western blotting has shown that gliadins, glutenins and proteins with similar molecular weights as the endogenous water/salt soluble wheat enzyme inhibitors are important allergens. Alpha and fast omega- are the most allergenic gliadins. The water/salt insoluble proteins share cross-reacting epitopes with water/salt soluble proteins. These data show that the numbers of proteins involved in the development of cereal hypersensitivity is greater than previously believed and that the development of specific IgE to alpha-gliadin may in part depend on the presence of cross-reacting antibodies to water/salt soluble flour allergens.

Antibodies, Blocking↗

Comparison of a radioallergosorbent (RAST) inhibition method and a monoclonal enzyme linked immunosorbent assay (ELISA) for aeroallergen measurement.

BACKGROUND: Mouse and rat urinary proteins are potent occupational allergens for exposed personnel. Methods of measuring airborne allergens differ greatly, and reported levels of allergens vary considerably between laboratories. OBJECTIVES: To compare the values obtained using two different methods of allergen detection. METHODS: Air samples were collected in rat rooms in Sweden and the United Kingdom at 2 L/min on to polytetrafluoroethylene (PTFE) filters and extracted in buffer containing 0.5% v/v Tween 20. Airborne rat urinary allergen (RUA) was measured in all samples by both RAST inhibition using a polyclonal human serum pool (UK) and a two monoclonal antibody sandwich ELISA employing antibodies specific for Rat n 1.02 (alpha2u-globulin) (Sweden). RESULTS: The two methods gave values which were correlated (r2 log values = 0.72, P<0.0001), but differed by several orders of magnitude (median [range] ratio of RAST inhibition/ELISA = 316 [7-26(80)]. There was a systematic bias: as the absolute values increased, the difference in the measurements increased. The rat urine standards used were antigenically similar. CONCLUSIONS: A large contrast in RUA values obtained from the two assays was observed in this study. This may be primarily due to methodological differences, but variations in antibody specificities or composition of allergenic epitopes in the air samples may contribute. The results demonstrate that standardization of methods and antibodies is necessary before interlaboratory comparisons can be made.

Air Pollutants, Occupational↗

Analysis of the allergenic composition of rat dust.

BACKGROUND: Allergy to rats is an important occupational health problem. The allergens of rat urine have been well defined but those in rat room dust, a potentially important source of inhalant exposure, have not. OBJECTIVE: To describe the allergens present in rat room dust and to identify a suitable marker protein which may be used to quantify airborne rat allergen. METHODS: Dust collected from the air-conditioning system (bulk dust, 'bd') and with an air sampler (airborne dust, 'ad') were analysed by radioallergosorbent test (RAST) inhibition, immunoblotting and immunoblot inhibition techniques and comparisons made with hair and urine extracts prepared from adult male Wistar rats. RESULTS: Extensive crossreactivity was found between the extracts by RAST inhibition under different experimental conditions. Dust was more potent as an inhibitor than other extracts. The immunoblotting patterns of both dusts were similar although 'ad' contained an allergen at 29 kDa not found in 'bd'. Forty-two sera from rat allergic subjects were used to identify 18 allergens in 'bd'. Three 'major' allergens were found; 100% of subjects had immunoglobulin (Ig)E to a 44 kDa allergen and 74% and 88% of subjects had IgE with bound to the 20.5 and 17 kDa allergens respectively. Immunoblot inhibition experiments identified the 17 kDa dust allergen as alpha 2u-globulin (Rat nI). CONCLUSIONS: Rat dust is a complex allergenic source. The 17 kDa dust allergen has immunological identity with Rat nI and is a suitable marker protein for the quantitation of airborne rat allergen.

Air Pollutants↗

Variation in rat urinary aeroallergen levels explained by differences in site, task and exposure group.

We describe how much of the variation in rat urinary aeroallergen (RUA) levels may be explained by differences in site and exposure group in research establishments and we identify task categories associated with high RUA levels. In this population 73% of the variability in RUA levels could be explained by exposure group, site and their interaction term. Exposure group alone explained the majority of the variation (69%). The task category associated with the highest RUA level was cleaning out, followed by indirect contact with rats and handling rats. The task categories associated with the lowest levels were slide production, post mortem and experiments. These results may help choose appropriate exposure measures for epidemiological studies in research establishments in the future and may help reduce the incidence of laboratory animal allergy.

Allergens↗

Identification of crossreacting wheat, rye, barley and soya flour allergens using sera from individuals with wheat-induced asthma.

We have conducted radio allergosorbent test (RAST), competitive RAST inhibition, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting using sera from patients with wheat-induced asthma to investigate the immunological relationship between wheat, rye, barley and soya, and to identify common proteins between these flours. RAST showed strong associations between the levels of specific IgE to wheat flour and those of rye and barley flour. Competitive RAST inhibition showed that wheat, rye, barley and soya flours contained crossreacting proteins, in decreasing concentrations. Wheat, rye and barley flours had similar protein profiles on gel electrophoresis. Soya flour contained a number of high molecular weight proteins not present in the other cereals. Western blotting using sera from 21 wheat flour hypersensitive individuals identified a large number of allergens in the different flours. Proteins of 69, 33, 26, 21 and 12 kDa were identified as major wheat flour allergens. Rye flour proteins of 21 and 12 kDa, and barley flour proteins of 69, 52 and 10 kDa were the major allergens identified by serum from wheat hypersensitive individuals. The major common protein of soya and wheat flour had a molecular weight of 21 kDa. The majority of crossreacting allergens identified between the different flours have molecular weights similar to those of known flour enzymes or enzyme inhibitors.

Allergens↗

Peak exposure concentrations of dust and flour aeroallergen in flour mills and bakeries.

As part of an epidemiological study amongst workers exposed to flour we measured peak exposure levels to total dust and flour aeroallergen with personal samplers in bakeries, flour mills and packing stations. Short-term tasks which were expected to give rise to high concentrations of exposure (peaks) were identified. The frequency and duration of these tasks were estimated and their levels of exposure to dust and flour aeroallergen measured. In total 209 samples were taken. The highest exposure concentrations both for dust (geometric mean > 30 mg m-3) and for flour aeroallergen (geometric mean > 500 micrograms m-3) were measured during certain operations. Exposure concentrations for the tasks were often much higher than the levels we had measured over a shift in a previous study. This might be important for sensitization and for the development of asthma. Peak exposure concentrations could be used to explore the exposure-response relationship more comprehensively. In general average flour aeroallergen concentrations increased linearly with average dust concentrations, although there were some exceptions.

Allergens↗

Measurement of airborne proteins involved in Bakers' asthma.

We have developed a competitive inhibition immunoassay to measure airborne flour, using purified polyclonal rabbit IgG antibodies to cereal flour antigens. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed different flours contained proteins of molecular weight similar to those present in air sample eluates. Western blotting demonstrated that the purified rabbit IgG antibodies bound to proteins whose molecular weights are similar to human allergens. Air sample eluates obtained from a large bakery had slopes of inhibition parallel to cereal flour when compared in the immuno-inhibition assay. The assay identified airborne proteins from several flours with a detection limit of 1 microgram ml-1. Using this immunoassay we have shown that a wide range of airborne flour concentrations occur in flour mills and a smaller range bakeries. This assay which allows measurement of the concentration of flour aeroallergen in the workplace will be valuable both in epidemiological studies and in examining the effectiveness of interventions to reduce the concentration of flour in the air.

Air Pollutants↗

The role of cereal and fungal amylases in cereal flour hypersensitivity.

To investigate the role of cereal alpha and beta-amylase in bakers' asthma, we have compared the IgE response of 30 wheat-flour-allergic individuals to barley alpha and beta-amylases with that of fungal alpha-amylase using radioallergosorbent test (RAST), RAST inhibition assays and Western blotting. RAST analysis showed 29 of the 30 subjects with inhalant induced cereal allergy had positive IgE to cereal amylases, but only 16 were positive to fungal alpha-amylase. Regression analysis showed an association between specific IgE to wheat-flour and to barley alpha-amylase (r = 0.70) and barley beta-amylase (r = 0.92) but a poor association with fungal alpha-amylase (r = 0.34). RAST inhibition showed minimal crossreactivity between barley alpha or beta-amylase and barley and fungal alpha-amylase. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting showed that non-reduced barley alpha-amylase had a molecular weight of 54 kDa and barley beta-amylase a molecular weight of 64 kDa. Reduced fungal alpha-amylase had a molecular weight of 54 kDa. Cereal alpha and beta-amylase appear to be important allergens in patients with allergy to flour.

Air Pollutants, Occupational↗