Sensitization to horse hair, symptoms and lung function in grooms

OBJECTIVE: This study aimed to investigate the rate of occupational sensitization to horse hair in grooms and whether occupational exposure to horse hair increases respiratory and allergic symptoms and affects lung function in grooms or not. METHODS: This is a cross-sectional study. Two hundred grooms were randomly selected among 1000 grooms working in Veliefendi Hippodrome of Istanbul. One hundred and twenty-five subjects agreed to enter the study. Ninety-two workers who worked in the different parts of this hippodrome enrolled as the control group. A detailed questionnaire including respiratory and allergic symptoms was filled in, physical examination, skin prick tests and pulmonary function tests were performed. RESULTS: Sensitization to horse hair was 12.8% in grooms and 4.3% in controls. The difference was statistically significant (P = 0.0035). Asthma was found in 14.4% of the grooms and 5.4% of the controls, allergic rhinitis in 42.4% of the grooms and 18.4% of the controls, allergic conjunctivitis in 35.2% of the grooms and 15.2% of the controls, and allergic skin diseases in 32.8% of the grooms and 13% of the controls. The differences were statistically significant (P = 0.043, P = 0.0002, P = 0.001 and P = 0.0008, respectively). The means of FEV1, FEV1/FVC and FVC parameters were significantly lower in the groom group (P = 0.006, P = 0.001 and P = 0.003, respectively). In the multivariate analysis, being in the groom group and working years were found to be predictive factors for impairments of lung function (P < 0.001 and P = 0.002, respectively). CONCLUSION: Occupational exposure to horse increases the sensitization to horse hair, induces asthma and allergic symptoms and also impairs lung functions.     

Evacuation of hematomas using liposuction technology: Technique and literature review

Established nonexpanding hematomas can be successfully treated with minimal morbidity using standard liposucstion techniques at the bedside or in an outpatient setting under local anesthesia. The authors presents a series of eight patients and discuss current concepts of dealing with this common and distressing surgical complication.     

Polymorphisms of the equine major histocompatibility complex class II DRA locus

The full extent of the polymorphism of ELA-DRA in Equidae is not yet known. Given the apparent differences in DRA polymorphisms between Equidae and other species, the aims of this study were to more fully characterize ELA-DRA, determine the extent of gene polymorphism and establish the allele-frequency distribution. An allele reference panel for the second exon of ELA-DRA was established by sequence-based typing of 69 equine DNA samples consisting of various breeds of domestic horse (Equus caballus), together with donkeys (Equus asinus), Grant's zebras (Equus boehmi) and one onager (Equus hemionus). Five of the six previously reported alleles detected using single-strand conformation polymorphism were found: ELA-DRA*0101, ELA-DRA*0201, ELA-DRA*0301, ELA-DRA*0501 (Albright-Fraser DG et al. Polymorphism of DRA among equids. Immunogenetics 1996: 43: 315-7) and ELA-DRA*0601 (GenBank accession number AF5419361). In addition to the previously reported alleles, five novel ELA-DRA alleles were detected within the ELA-DRA allele reference panel. One of these was identified in E. caballus (ELA-DRA*JBH11), one in E. boehmi and E. hemionus (ELA-DRA*JBZ185) and three in E. asinus (ELA-DRA*JBD3, ELA-DRA*JBD17 and ELA-DRA*JBH45). A total of 565 equine DNA samples were screened using reference-strand-mediated conformation analysis, a double-stranded conformation-based mutation detection system that can be used to type existing ELA-DRA alleles and identify new variants. Based on our findings, at least 11 ELA-DRA alleles are now known to exist, and this level of polymorphism at the DRA locus appears to be unique to the genus Equus. Both the previously reported alleles and the new alleles displayed a species-specific distribution.     

蝰蛇毒压电免疫传感器固定方法的研究

目的：为研制蝰蛇毒压电免疫传感器，研究抗蛇毒抗体固定于石英晶体银电极表面的固定技术。方法：采用马抗蝰蛇毒血清抗体和抗蝰蛇毒鸡卵黄抗体作为生物敏感材料，对比研究了胱胺自组装-PSS反相吸附法和PEI粘附-戊二醛交联法：比较了采用两种固定方法所制的压电免疫传感器的性能。结果：鸡卵黄抗体采用PEI粘附-戊二醛交联法效果较好，其制备的IgY压电免疫传感器检测蝰蛇毒灵敏度为0．5ug／mL；而马血清抗体用胱胺自组装-PSS反相吸附法较好，其制备的IgG’免疫传感器检测蝰蛇毒灵敏度为10ug／mL。结论：以PEI粘附-戊二醛交联法固定抗蝰蛇毒鸡卵黄抗体所制备的蝰蛇毒压电免疫传感器的性能稳定，特异性好，可实现蛇毒的快速检测。     

The enhancement of antibody response by IgM antibodies is dependent on antigen-specific T helper cells

The enhancement of antibody responses by IgM antibodies administered with low doses of antigen has been studied in a T-dependent (SRBC) and an T-independent (alpha 1,6 dextran) system. It has been found that IgM anti-SRBC antibodies do not enhance a SRBC response in nude mice. The T-cell dependency was also directly demonstrated by showing the effect of IgM on T-cell priming in transfer experiments. The simultaneous injection of antigen and IgM antibody also induced a polyclonal increase of IgM, PFC, which was not due to a non-specific "adjuvant" effect of IgM, as we could not detect a similar effect on an ongoing response to HRBC in mice simultaneously given SRBC and IgM anti-SRBC antibodies. The specificity of the helper cell for either the antibody or the antigen was investigated in a response to alpha 1, 6 dextran, in which we could demonstrate antibody-specific helper T cells, but no antigen-specific help. We have found that IgM anti-dextran antibodies do not enhance and rather suppress the response of normal, high-responder mice, to dextran, suggesting that the T cells mediating the "19S enhancement" are antigen-specific. The magnitude of the enhancement response, as compared to the responses induced by either antigen or antibody alone, implies a synergistic mechanism, possibly involving antigen-specific and antibody(idiotype)-specific T helper cells.     

Dispersion of horse allergen in the ambient air, detected with sandwich ELISA

BACKGROUND: The objective was to establish an ELISA to detect horse allergen in ambient air and settled dust. METHODS: Monoclonal antibodies (mAbs) were produced against extracts of horse antigen. Two mAbs were selected and used in a sandwich ELISA. By the aid of portable pumps, air samples were collected in one stable and in the ambient air surrounding this stable. Furthermore, settled dust was collected by wiping spots with pieces of fabric, at sites within 500 m of the stable. RESULTS: Extracts of horsehair could be extensively diluted and still be positive. Extracts of cat and dog allergen failed to be detected. Furthermore, the mAbs were shown to detect an IgE-binding component. This was demonstrated by an ELISA using mAbs as capture antibody and sera from horse-allergic subjects as secondary antibody with readout depending on anti-IgE antibody. The sera with the highest RAST class to horse were positive in this ELISA. Airborne levels of horse allergen were over 500-fold higher in the stable than just outside the stable and over 3000-fold higher than at a residential building located only 12 m from the stable. Similarly, an inverse correlation was found between the distance to the stable and levels of "outdoor settled" horse allergen (r=-0.9, P<0.001). CONCLUSION: We have developed a sensitive, horse-allergen-specific, mAb assay allowing detection of low levels of horse allergens. Raised levels of horse allergen were found outdoors only in the close vicinity of the stable.     

Study of cobalt doping control via various routes in thoroughbred horses

Cobalt is a substance that has been abused for athletic performance enhancement and has thus been prohibited by human and animal sports doping control authorities. However, because cobalt is present in humans and animals as a trace element, a certain level of cobalt is naturally present in their excretions. In the racing industry, cobalt is a controlled substance with a threshold concentration specified by the International Agreement on Breeding, Racing and Wagering (IABRW) for international harmonization. Due to environmental and feed consumption differences among countries, regional cobalt concentration trends should be evaluated before cobalt testing is introduced. In this study, we conducted a preliminary evaluation of the urinary concentration of cobalt among a population of racehorses in Korea using inductively coupled plasma mass spectrometry (ICP-MS) analysis, followed by analysis of the urinary release of cobalt after the administration of cobalt chloride in various situations. The normal distribution for the Korea-based racehorses was used to determine a urine concentration limit (96.5 ng/ml, risk factor of 1 in 10,000). After the intravenous (IV) administration of CoCl₂, the initial elimination of cobalt was rapid. A high concentration (over 2,000 ng/ml) and a slow excretion pattern were observed during the final 2 weeks of the 3-week observation period. When CoCl₂ was administered orally, maximum concentration (C_max, 92–992 ng/ml) was observed at 6–8 h.     

Direct sequence confirmation of qPCR products for gene doping assay validation in horses

The misuse of gene therapy by the introduction of transgenes via plasmid or viral vectors as a doping agent is an increasing concern in human and animal sports, not only in consideration to fair competition but also in potential detrimental effects to welfare. Doping events can be detected by polymerase chain reaction (PCR) amplification of a transgene-specific region of DNA. Quantitative real-time PCR (qPCR) is particularly suited to confirmatory investigations where precise limits of detection can be calculated. To fully validate a qPCR experiment, it is highly desirable to confirm the identity of the amplicon. Although post-PCR techniques such as melt curve and fragment size analysis can provide strong evidence that the amplicon is as expected, sequence identity confirmation may be beneficial as part of regulatory proceedings. We present here our investigation into two alternative processes for the direct assessment of qPCR products for five genes using next-generation sequencing: ligation of sequence-ready adapters to qPCR products and qPCR assays performed with primers tailed with Illumina flow cell binding sites. To fully test the robustness of the techniques at concentrations required for gene doping detection, we also calculated a putative limit of detection for the assays. Both ligated adapters and tailed primers were successful in producing sequence data for the qPCR products without further amplification. Ligated adapters are preferred, however, as they do not require re-optimisation of existing qPCR assays.