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1.
Atopic dermatitis in dogs is a common allergic skin disease that affects substantial numbers of dogs in the UK. The purpose of this study was to compare the results of an intradermal test (IDT) and an in vitro test in a large cohort of dogs. Dogs were intradermal tested with Greer allergens (Greer Labs Inc, Lenoir, NC, USA) using standard techniques. At the same time blood samples were drawn and submitted for evaluation by ELISA using the ALLERCEPT Definitive Allergen Panels for allergen-specific IgE, a commercial assay that uses a biotinylated recombinant extracellular domain of the high affinity Fc-epsilon receptor alpha chain protein (Fcepsilon RIalpha). The allergens used in the two tests included grass, tree and weed pollens, moulds, flea saliva/whole flea extract and house dust mite species. The optical density readings from the ELISA for each allergen were compared with the results of the IDT for 265 dogs. The prevalence of positive reactions in the ELISA was equal to or greater than the results of the IDT in the case of almost all of the allergens, but two notable exceptions were the house dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus. These two allergens were the most common positive reactions by IDT (prevalence D. farinae 78.9%, D. pteronyssinus 66.4%). The results of the two tests were significantly different (McNemar's test, P<0.05) for 16 of the 22 allergens. The sensitivities of the ELISA compared to the IDT (where there were more than 3 dogs with positive reactions in both tests) varied between 19.3 and 77.1% (D. pteronyssinus 19.3% and D. farinae 67.9%) and the specificities varied between 64.2 and 96.6% (D. pteronyssinus 96.6% and D. farinae 89.3%).  相似文献   

2.
OBJECTIVE: To compare a radioallergosorbent test and 2 ELISA with intradermal testing for the determination of environmental allergen hypersensitivity in horses with and without atopic diseases. DESIGN: Prospective clinical study. ANIMALS: 10 horses with recurrent urticaria, 7 with atopic dermatitis, 16 with chronic obstructive pulmonary disease, and 22 without atopy. PROCEDURE: History, physical examination, hemogram, serum biochemical analyses, bronchoalveolar lavage, and an intradermal test (used as the criterion standard) with a regional panel of 73 allergens were performed in all horses. Serum was analyzed by use of the 3 in vitro assays of allergen-specific IgE. RESULTS: An ELISA based on the alpha chain of the high-affinity IgE receptor, the Fcepsilon receptor immunoglobin epsilon chain (FcepsilonRIalpha) for IgE, had the overall highest kappa statistic (0.238), positive predictive value (49%), and negative predictive value (78%). Overall agreement between the FcepsilonRIalpha-based ELISA and the intradermal test was fair. The highest kappa statistic was obtained by the FcepsilonRIalpha-based ELISA in horses with atopic dermatitis (0.330). Kappa statistics for the radioallergosorbent test and a polyclonal antibody-based ELISA agreed slightly with that of the intradermal test at best. CONCLUSIONS AND CLINICAL RELEVANCE: None of the 3 serum allergy tests reliably detected allergen hypersensitivity, compared with the intradermal test. The FcepsilonRIalpha-based ELISA performed significantly better overall than the other 2 tests. Low sensitivity of all 3 assays indicates the need for continued study to elucidate a more sensitive test for the determination of potentially pathogenic allergens in horses.  相似文献   

3.
The importance of pollen allergies in dogs with atopic dermatitis in South America has not been determined. Local allergists and pallinologists have performed a few studies evaluating pollen counts in Buenos Aires throughout the year. Those studies helped the author in choosing a pollen allergen panel for intradermal testing based on local allergens. One-hundred-sixty dogs with atopic dermatitis were tested intradermally during a 3-year period, using 30 individual allergens including house dust mites, moulds, trees, weeds and grasses. The most important pollen allergens in terms of positive reactions were Platanus acerifolia (32 dogs), Fraxinus Americana (11 dogs), Cynodon dactylon (13 dogs), Ambrosia tenuifolia (26 dogs), Artemisia spp. (16 dogs), Plantago lanceolata (21 dogs), Chenopodium spp. (16 dogs) and grass pollen (29 dogs). Other pollens on the panel were less important. A correlation between pollination season and atopic dermatitis symptoms was determined. A few dogs were only positive for pollen and not for house dust mites. The incidence of pollen allergies in canine atopic dermatitis in South America must be determined in order to select an adequate panel for the area and for its main cities, and to exclude those allergens that are irrelevant locally. Special individual cases might be further tested, if necessary, with other allergens upon plant identification in the patient's environment. The findings encourage more investigation in this area, and suggest that allergen-specific immunotherapy with tree, weed and grass pollen should be considered.
Funding: Self-funded.  相似文献   

4.
Equine disease with an allergic etiology is common. Environmental antigens most often implicated as allergens in horses include molds, dusty hay, grass pollen, hay dust mites, and insect saliva. Although intradermal testing with allergen is a useful diagnostic tool for some species, skin testing frequently produces false positive results in horses. Allergen deprivation as a diagnostic tool is often impossible and at best it is ineffective at diagnosing the specific allergic reactivity. Synthesis of IgE after exposure to allergen is the instigator of the allergic process. While IgE exerts its effect after binding strongly to mast cell Fc receptors, the presence of free IgE in the serum can be used to quantify and determine the allergen specificity of the allergic disease. A lack of widely available reagents for detection of equine IgE has limited this approach in horses. We have used the nucleotide sequence of equine IgE to prepare a peptide-based immunogen to elicit equine epsilon chain-specific antisera. Selection of peptides was based on antigenic attributes of the deduced amino acid sequence of the equine epsilon chain. Six peptides were selected for conjugation to carrier molecules and rabbit immunization. Of these, one peptide elicited antisera that was successfully used in enzyme linked immunosorbant assay (ELISA) to screen horse serum from 64 allergic horses for allergen-specific IgE. Twenty-four of the 64 horses showed positive reactivity to one or more of the following allergens: grass, grain mill dust, mosquito, and horsefly. This study demonstrates the usefulness of peptide-based immunogens for development of antisera to rare or difficult to purify antigens such as IgE. Resultant antisera has great usefulness in diagnostic assays for equine allergy and as a research tool.  相似文献   

5.
Twenty-eight atopic dogs, 22 pruritic, non-atopic dogs and 10 healthy dogs were ELISA tested. For calculations of diagnostic specificity and sensitivity, positive ELISA test results in non-atopic dogs were considered false positive results. The absence of any positive results in the atopic dogs was considered false negative results. The atopic dogs were tested both with ELISA and an intradermal test, utilising allergen extracts from the same manufacturer, to determine the frequency of positive allergen reactions in the ELISA test compared with the intradermal test. The Prausnitz-Küstner test was performed to evaluate the significance of a positive ELISA test result. Based on cross-tabulations with clinically defined atopic dermatitis, the ELISA test showed a sensitivity of 53.6% and a specificity of 84.4%. The correlation between the ELISA and the intradermal test was poor. Positive Prausnitz-Küstner tests were not obtained using sera from dogs that were intradermal test negative for the tested allergens, even though sera had high levels of IgE as measured by the ELISA. These findings question the significance of a positive ELISA test result and indicate that the test is not measuring functional allergen-specific IgE.  相似文献   

6.
The purpose of this study was to define the operational and performance characteristics of a commercially available monoclonal antibody based (mac) ELISA for detection of allergen-specific IgE in dogs. The average intra-assay variance over 1 year was 9.7% (range 2.5–62.7%), while the interassay variance averaged 10.8% (range 8.1–13.8%). The average positive control responses observed for grass, weed, tree and mite allergens during each month remained relatively constant; the average monthly variance was 11.6% (range 8.3–19.2%) for grass pollens, 13.3% (range 9.1–20.4%) for weed pollens, 13.3% (range 9.8–18.2%) for tree pollens and 13.6% (range 8.9–18.7%) for mite allergens. The interlaboratory concordance of results for the macELISA was approximately 91%. The interlaboratory concordance of results comparing the macELISA and a high affinity IgE receptor-based ELISA was approximately 92%. The results demonstrate that the macELISA is reproducible and the results are comparable to the high affinity IgE receptor based ELISA within and between laboratories.  相似文献   

7.
Skin hypersensitivities (allergies) in horses are often diagnosed using clinical signs only. Intradermal testing or serological assays are diagnostic options to confirm the allergic nature of the disease and to identify the allergen(s). Our objective was to develop an allergen-specific enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody specific for horse IgE and to examine its potential for allergen detection in serum in comparison to intradermal testing. Intradermal testing with 61 allergen extracts was performed on 10 horses affected with skin hypersensitivity. Their sera were analyzed by ELISA for IgE antibodies to the same allergens. The kappa test of concordance was used for comparison of the results of both tests. Out of 61 allergen extracts, only two (Timothy and Quack) had kappa values greater than 0.60, suggesting a substantial agreement between skin testing and IgE ELISA. The statistical comparison of the remaining 59 allergens showed little or no concordance between the tests beyond chance. To identify parameters that may influence the sensitivity of the ELISA, the assay was modified to detect allergen-specific IgGb and IgG(T) in serum, and the protein content in all allergen extracts was determined by SDS-PAGE. The commercial allergen extracts revealed a high variation in detectable protein. High concentrations of allergen-specific IgG in horse serum were found to compete with IgE for binding to the plates. In conclusion, an ELISA using whole serum and crude allergen preparations provides limited diagnostic information in horses. The reliable diagnosis of allergens in equine skin hypersensitivity is essential to improve allergen-specific treatments, such as hyposensitization, or the development of allergy vaccines.  相似文献   

8.
This study evaluated a new perspective in the diagnosis of dermatitis in dogs with signs suggestive of allergic skin disease. The results obtained with CMG IMMUNODOT tests using the technique of allergen-specific strip tests, as employed for human allergy diagnosis, were compared with those obtained by the intradermal skin test (IDST). Forty-eight cases completed the diagnostic evaluation, which included IDST, flea-control program, exclusion of sarcoptes and, for some cases, a 1- to 2-month stabilization period on a restricted protein source diet and testing the serum in the presence of allergen-specific IgE and total IgE. The most common disorders included house and storage dust mites, allergic dermatitis and flea-allergic dermatitis together with atopy. This was confirmed serologically. In the case of positive IDST to pollens, Aspergillus spp. and cat epithelium, CMG IMMUNODOT strip tests were negative. A total of 25% of cases were considered to be primarily associated with food hypersensitivity, but only 4% were confirmed serologically. This study emphasizes the value of CMG IMMUNODOT tests as a support in the diagnosis of dog allergy.  相似文献   

9.
The aim of this study was to compare the results of an intradermal skin test (IDST) with those of an allergen-specific IgE-ELISA in 210 dogs with atopic dermatitis. All the dogs had a clinical diagnosis of atopic dermatitis and underwent an IDST. The sera of all dogs were analysed for allergen-specific IgE by ELISA using the monoclonal antibody D9 against dog IgE. IDST was used as the standard assay. In both methods, the following antigens provided a positive test result: Dermatophagoides farinae, Acarus siro, Tyrophagus putrescentiae, ragweed, mugwort and Lepidoglyphus destructor. ELISA had an overall sensitivity of 82.4% and an overall specificity of 93.8%. The overall accuracy of the ELISA was 91.3%. The evaluated monoclonal D9 ELISA was found to be a reliable tool for the diagnosis of those allergens that cause clinical atopy, and can be recommended for use in dogs when immunotherapy is a therapeutic option.  相似文献   

10.
In vitro assays for allergen specific immunoglobulin E (IgE) are a convenient and reproducible alternative to intradermal skin testing in dogs. Such tests may be used to support a diagnosis of atopic dermatitis and to define appropriate allergens for immunotherapy. Current in vitro assays rely upon monoclonal or polyclonal antibodies as IgE detection reagents. However, in sera where allergen-specific IgG occurs in great excess, any IgE:IgG cross-reactivity of the detection reagent may result in lowered assay specificity. Therefore, we have developed an assay for canine IgE which uses a recombinant form of the extracellular part of the alpha chain of the human high affinity IgE receptor (FcvarepsilonRIalpha). Biotinylated FcvarepsilonRIalpha shows no significant binding to purified canine IgG, and recognizes a heat labile antibody in serum, with a detection limit of 73-146pg/ml. Comparison of assay signals using the labeled FcvarepsilonRIalpha and a highly specific anti-canine IgE monoclonal antibody (MAb) shows good agreement. The FcvarepsilonRIalpha is therefore a sensitive and specific alternative to polyclonal or monoclonal antibodies for canine serum IgE measurement.  相似文献   

11.
The success of the treatment of 117 dogs with atopic dermatitis with allergen-specific immunotherapy for up to 48 months was assessed. An excellent response (remission with exclusive immunotherapy) was recorded in 18 of the dogs, a good response (more than 50 per cent reduction in medication and improvement of clinical signs) was recorded in 57, a moderate response was recorded in 24 and a poor response in 18. The mould antigens in the allergen extract were stored in a separate vial before administration and the success rate of the immunotherapy including mould antigens was much higher than in an earlier study in which mould and pollen antigens had been stored in one vial. The success rate was not affected significantly by the age of the dogs when the disease developed, or by their age or the period for which they had shown clinical signs when the treatment began; it was also unaffected by whether pollens, moulds or dust mites were used as antigens, or by whether the offending allergens had been identified by intradermal testing or by serum testing for allergen-specific immunoglobulin E.  相似文献   

12.
The objective of this study was to evaluate the accuracy of in vivo and in vitro tests in the diagnosis of flea allergy dermatitis in comparison with history, clinical signs and response to flea control. Intradermal testing using four different sources of flea allergens and FcepsilonRIalpha-based immunoglobulin (Ig)E assays were performed in 15 flea-allergic dogs, 15 atopic dogs and 15 dogs infested with fleas but showing no clinical signs of skin disease. Sensitivity, specificity, negative predictive value, positive predictive value and accuracy were calculated for all five tests and results varied greatly. Sensitivity, specificity and overall accuracy were 27, 83 and 64%, respectively, for one extract (Isotec), 67, 90 and 82% for another extract (Greer), 93, 90 and 91% for flea saliva, 40, 90 and 73% for the recombinant Cte f 1 both produced by Heska Corp. and 87, 53 and 64% for a FcepsilonRIalpha-based IgE assay. These results indicate that intradermal testing with flea extracts is more accurate in the diagnosis of flea allergy dermatitis than in vitro tests. Moreover, pure flea saliva used as a reagent for intradermal testing provided the best results in terms of sensitivity, specificity and overall accuracy although the Greer extract, a whole body flea extract, also allowed a good correlation between intradermal testing results and clinical approach to flea allergy dermatitis diagnosis.  相似文献   

13.
OBJECTIVE: To determine the most relevant aero-allergens involved in canine atopic dermatitis in southeastern Australia and provide information about these aero-allergens to the general practitioner. PROCEDURE: Dogs presented to the Animal Skin & Allergy Clinic with history and clinical signs of atopic dermatitis were injected intradermally with 38 different allergens and negative and positive control. Intradermal skin tests in 1000 dogs were retrospectively evaluated. RESULTS: One third of all patients reacted to the house dust mite Dermatophagoides farinae. Allergens reacting in more than 15% of the patients were wheat (Triticum aestivum), sweet vernal (Anthoxanthum odoratum), English couch (Agropyron repens), yellow dock (Rumex crispus), Mexican tea (Chenopodium ambrosioides), plantain (Plantago lanceolata), melaleuca (Melaleuca quinquenervia) and peppercorn (Schimus spp). CONCLUSION: House dust mites are the most common allergens in canine atopic dermatitis in southeastern Australia and D farinae is involved most frequently. However, a number of grass, weed and tree pollens also are involved regularly.  相似文献   

14.
The purpose of this study was to determine whether cats with allergic skin disease have significant concentrations of serum Immunoglobulin E (IgE) specific for antigens derived from the house dust mites (HDM) Dermatophagoides farinae (DF) and Dermatophagoides pteronyssinus (DP). Enzyme-linked immunosorbent assays (ELISA) were developed for this purpose. Binding of serum allergen-specific IgE was detected via the use of biotinylated Fc-epsilon receptor alpha chain protein (FcvarepsilonRIalpha). Following optimisation of the assay, serum samples from 59 cats with allergic skin disease and 54 clinically normal cats were screened. Results were expressed as ELISA units per ml (EU/ml) compared to a standard curve. Serological findings were correlated with the clinical presentation of affected cats. Cats with symptoms of feline allergic skin disease were grouped as follows: self-induced alopecia without lesions (group 1), papulocrusting dermatitis (group 2), eosinophilic granuloma complex (group 3), papular/ulcerative dermatitis of head and neck/facial dermatitis (group 4), and a combination of symptoms (group 5). Control normal cats comprised the final group (group 6). The Kruskal-Wallis test was used for statistical analysis. There was no significant difference between groups for DF- and DP-specific IgE concentrations with a p-value of 0.875 and 0.705, respectively. Although the FcvarepsilonRIalpha-based ELISA was able to detect house dust mite-specific feline IgE, the presence of this allergen-specific IgE correlates poorly with the presence of clinical manifestations of allergic skin disease. The results of this study question the clinical relevance of house dust mite-specific IgE in feline allergic skin disease.  相似文献   

15.
The availability of anti-dog IgE monoclonal antibodies has enabled development of highly specific ELISA assays for measuring antigen-specific IgE in dog serum. In this article the authors propose criteria for evaluation of these monoclonals and demonstrate that some anti-human IgE monoclonals recognize dog IgE. Combinations of two or more monoclonal antibodies can enhance assay sensitivity; for example, a mixture of DE38.HRPO and 4F4.HRPO conjugates detect total dog IgE in the range 10–10000 ng/mL. Results are reported from nine clinical studies conducted in Europe, Japan and Australia involving more than 400 dogs in which serologic IgE determinations performed using the CMG IMMUNODOT strip system for house dust mites, storage mites, flea, grass pollens and moulds were compared with immediate skin test findings. House dust mites were identified as the common major dog allergen throughout these areas although regional reactions to food allergens were observed. These results confirm that the CMG IMMUNODOT system is a valuable and reliable diagnostic test for dog allergy.  相似文献   

16.
In humans with atopic dermatitis and in mouse models of IgE-mediated allergic diseases, evidence is mounting that the stratum corneum (SC) provides an important barrier against environmental allergens. At this time, it is not known whether the SC has a similar role in dogs, especially in those with atopic dermatitis. The objectives of this pilot study were to determine whether SC removal led to earlier and stronger sensitization of atopic dogs to Dermatophagoides farinae (Df) house dust mites. Five Maltese-beagle atopic (MBA) dogs were sensitized epicutaneously after the SC was removed with ten tape strips (TS group), while sensitization was done without tape strips in five other MBA dogs (nontape stripping; NTS group). During this 16 week study, sensitization was assessed with allergen-specific IgE serology, intradermal testing with Df allergens and determination of stimulation indices of blood mononuclear cells cultured with Df and stained for CD4 and the activation markers CD25 or CD30. Compared with dogs from the NTS group, those of the TS group exhibited earlier rises in Df-specific IgE serum levels, usually had higher allergen-specific IgE titres, showed higher intradermal test reactivity and had earlier increases and higher percentages of CD25- or CD30-positive activated allergen-specific peripheral CD4-positive T lymphocytes. These observations implicate a role of the SC as a barrier limiting sensitization to exogenous allergens in this experimental atopic dog model.  相似文献   

17.
The intradermal skin test (IDST) and serologic allergy test (SAT) has been developed for confirming a diagnosis of canine atopy and determining allergens for immunotherapy. To determine the prevalence of causative allergens for canine atopic dermatitis in Japan, IDST and SAT were performed with the CMG Immunodot strips on 95 atopic dogs using 9 allergens. In addition, we compared agreement rate, sensitivity and specificity between them (using IDST as the standard). The allergen most commonly positive in both tests was house dust mites (IDST: 69.5%, SAT: 48.4%). Moreover, Japanese cedar, mugwort and grass mix were detected as attendant causative allergens. Agreement rates between the two tests ranged from 67.4% to 96.8%; the overall mean agreement rate were 81%. SAT was shown to have sensitivity to IDST ranging from 16.7 to 68.2%. The specificities were very high for all allergens, on the order of 94.9-100% (median=98.7%). Finally, the efficacy of immunotherapy was evaluated on 27 atopic dogs based on IDST (15 dogs) and SAT (12 dogs) results. Overall, 60% (9/15) of the IDST group and 66.8% (8/12) of the SAT group experienced a 50% to 100% reduction in their symptomatology. No significant differences were found in response to immunotherapy during the follow-up period between allergen selection methods. These results indicate the value of serologic tests as an aid to identifying an allergen solution for immunotherapy.  相似文献   

18.
OBJECTIVE: To evaluate a model for atopic dermatitis (AD) and to measure the effect of sensitization in Beagles genetically predisposed to produce high serum concentrations of allergen specific IgE. ANIMALS: 22 laboratory Beagles. PROCEDURE: Seventeen dogs were sensitized from birth to 3 allergens (recombinant birch pollen, Dermatophagoides pteronyssinus, and D farinae). Five nonsensitized dogs from the same litters served as controls. Clinical scoring, regular intradermal testing, measurement of serum concentrations of allergen-specific IgE, and collection of biopsy specimens of skin at 23, 32, and 43 weeks of age were performed. Serial tissue sections were stained for identification of IgE+ cells, mast cells and their subtypes, T-cells, Langerhans cells, and major histocompatibility complex class-II+ cells. At the age of 15 months, dogs were continuously exposed to 2 microg of mite allergen/g of dust. RESULTS: Sensitized dogs had positive intradermal test reactions and significantly higher serum concentrations of allergen specific IgE, compared with nonsensitized dogs. In sensitized and nonsensitized dogs, a significantly higher number of mast cells was found at predilection sites, compared with the control biopsy site. The number of mast cells at predilection sites increased with age. Sensitization significantly increased the number of epidermal Langerhans cells by 23 weeks of age. The number of epidermal Langerhans cells significantly increased in nonsensitized dogs by 32 weeks of age. Clinical scoring only revealed mild transient erythema in some dogs. CONCLUSIONS: increases in concentrations of serum allergen-specific IgE and exposure to allergens is not sufficient to induce clinical signs of AD in genetically predisposed dogs.  相似文献   

19.
Clinically important allergens for the diagnosis and treatment of atopic dermatitis vary geographically. In order to identify the most prevalent allergens in atopic dogs in Japan, 42 dogs with a clinical diagnosis of atopy were tested using both in vivo (intradermal skin test (IDST)) and in vitro (antigen-specific IgE assay) allergy tests. Allergens used for IDST included 26 allergen extracts from eight allergen groups: trees, weeds, grasses, house dust mites (HDM), molds, foods, epithelia, and arthropods. Immunodot assay was used to measure antigen-specific IgE against 24 allergens from these eight groups and against fish such as cod and sole. In the 42 dogs, the most common positive allergen reaction was to HDM on both IDST (29/42 dogs or 69%) and in vitro testing (23/42 or 54.8%). The second most frequent positive allergen reaction was to Japanese cedar pollen (21/42 or 50.0% for IDST and 7/42 or 16.7% for in vitro testing). In both tests, less than 20% of dogs had positive reactions to molds or foods. Positive reactions to cat epithelia were frequently found on IDST, but rarely found on in vitro testing. Agreement between the two tests was found in 26 instances: HDM (21 dogs), Japanese cedar pollen (five dogs) and wheat (one dog). In this study, the two most common allergens involved in atopic dermatitis in dogs in Japan were HDM and Japanese cedar pollen.  相似文献   

20.
OBJECTIVE: To determine the prevalence of serum IgE against the storage mites Acarus siro, Blomia tropicalis, and Tyrophagus putrescentiae in a population of dogs with atopic dermatitis. SAMPLE POPULATION: Sera from 84 dogs with atopic dermatitis residing in various regions of the United States and Europe. PROCEDURE: Immunoblotting of sera from atopic dogs was used to identify proteins in mite extracts that bound IgE. RESULTS: 94% of the dogs had serum IgE against proteins in extracts of 1 or more of the storage mite species. Ninety-five, 92, and 89% of the storage mite-sensitive dogs had serum IgE against proteins in extracts of A siro, B tropicalis, and T putrescentiae, respectively. Eighty-two percent had serum IgE against at least 1 protein in all 3 species. Most of the major allergens had molecular weights > 80 kd. A greater percentage of the dog sera had IgE against storage mite proteins, compared with proteins of the house dust mites Dermatophagoides farinae and D pteronyssinus. CONCLUSION AND CLINICAL RELEVANCE: Many dogs with atopic dermatitis have serum IgE against many allergens of storage mites. Most of these allergens, like allergens of dust mites, had molecular weights > 80 kd. Storage mite sensitivity in dogs may be as important, if not more important, than dust mite sensitivity.  相似文献   

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