An Immunoassay for Dibutyl Phthalate Based on Direct Hapten Linkage to the Polystyrene Surface of Microtiter Plates

BACKGROUND: Dibutyl phthalate (DBP) is predominantly used as a plasticizer inplastics to make them flexible. Extensive use of phthalates in both industrial processes and other consumer products has resulted in the ubiquitous presence of phthalates in the environment. In order to better determine the level of pollution in the environment and evaluate the potential adverse effects of exposure to DBP, immunoassay for DBP was developed. METHODOLOGY/PRINCIPAL FINDINGS: A monoclonal antibody specific to DBP was produced from a stable hybridoma cell line generated by lymphocyte hybridoma technique. An indirect competitive enzyme-linked immunosorbent assay (icELISA) employing direct coating of hapten on polystyrene microtiter plates was established for the detection of DBP. Polystyrene surface was first oxidized by permanganate in dilute sulfuric acid to generate carboxyl groups. Then dibutyl 4-aminophthalate, which is an analogue of DBP, was covalently linked to the carboxyl groups of polystyrene surface with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC). Compared with conjugate coated format (IC(50)=106 ng/mL), the direct hapten coated format (IC(50)=14.6 ng/mL) improved assay sensitivity after careful optimization of assay conditions. The average recovery of DBP from spiked water sample was 104.4% and the average coefficient of variation was 9.95%. Good agreement of the results obtained by the hapten coated icELISA and gas chromatography-mass spectrometry further confirmed the reliability and accuracy of the icELISA for the detection of DBP in certain plastic and cosmetic samples. CONCLUSIONS/SIGNIFICANCE: The stable and efficient hybridoma cell line obtained is an unlimited source of sensitive and specific antibody to DBP. The hapten coated format is proposed as generally applicable because the carboxyl groups on modified microtiter plate surface enables stable immobilization of aminated or hydroxylated hapten with EDC. The developed hapten coated icELISA can be used as a convenient quantitative tool for the sensitive and accurate monitoring DBP in water, plastic and cosmetic samples

Data on megakaryocytes in the bone marrow of mice exposed to formaldehyde.

Previously, we reported that occupational exposure to formaldehyde (FA) exposure in factory workers reduced platelet counts, http://dx.doi.org/10.1158/1055-9965.EPI-09-0762[1], while exposure in mice increased platelet counts http://dx.doi.org/10.1371/journal.pone.0074974[2]. Bone marrow megakaryocyte (MK) numbers were also increased in exposed mice, as determined qualitatively. The data presented here are from a quantitative evaluation of MK numbers in the bone marrow histopathological slides from the previous FA exposure experiments in mice. Bone marrow slides were prepared using a single 5 μm section of femur from 2 mice randomly selected from each exposure group (n=9) treated with 0, 0.5 and 3.0 mg/m(3) FA by nose-only inhalation. MKs were systemically counted and average MK frequency was calculated as the total MK per slide divided by the number of fields evaluated. Data are presented visually as microscopy views and graphically as MK frequency

Development of two enzyme-linked immunosorbent assay formats for thifluzamide residues’ analysis based on distinct polyclonal antibodies

Haptens 2-Methyl-4-(trifluoromethyl)thiazole-5-carboxylic acid and 2,6-Dibromo-4-(trifluoromethoxy)aniline, the two moieties of thifluzamide, were conjugated with carrier proteins for the synthesis of artificial antigens. Two distinct anti-thifluzamide polyclonal antibodies (PAb-1 and PAb-2) were produced from the immunized female Balb/c mice. The indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) in two formats based on the PAbs was developed for thifluzamide analysis. The concentration of 50% inhibition (IC50) of ELISA-1 was 1.39 mg L−1 and its limit of detection (LOD) was 0.082 mg L−1. Meanwhile, ELISA-2 had a similar IC50 of 1.96 mg L−1 and a LOD of 0.074 mg L−1 as ELISA-1. Both the raised PAbs exhibited high specificity to thifluzamide. The recoveries for spiked samples including water and wheat ranged from 72.0% to 128.4%, and the accuracy of ELISA was confirmed by high-performance liquid chromatography. In summary, the ic-ELISA might be a promising tool for simple, sensitive and rapid detection of thifluzamide residues in real samples

Evaluation of Drifting Snow Susceptibility Based on GIS and GA-BP Algorithms

Drifting snow, the flow of dispersed snow particles near ground level under the action of wind, is a major form of snow damage. When drifting snow occurs on railways, highways, and other transportation lines, it seriously affects their operational safety and results in drifting snow disasters. Drifting snow disasters frequently occur in the high latitudes of northwest China. At present, most scholars are committed to studying the prevention and control measures of drifting snow, but the prerequisite for prevention is to effectively evaluate the susceptibility of drifting snow along railways and highways to identify areas with a high risk of occurrence. Taking the Xinjiang Afukuzhun Railway as an example, this study uses a geographic information system (GIS) combined with on-site monitoring and surveys to establish a drifting snow susceptibility evaluation index system. The drifting snow susceptibility index (DSSI) is calculated through the weight of an evidence (WOE) model, and a genetic algorithm backpropagation (GA-BP) algorithm is used to obtain optimised evaluation index weights to improve the accuracy of model evaluation. The results show that the accuracies of the WOE model, WOE backpropagation (WOE-BP) model, and weight of evidence genetic algorithm backpropagation (WOE-GA-BP) model are 0.747, 0.748, and 0.785, respectively, indicating that the method can be effectively applied to evaluate drifting snow susceptibility

Data on megakaryocytes in the bone marrow of mice exposed to formaldehyde

Previously, we reported that occupational exposure to formaldehyde (FA) exposure in factory workers reduced platelet counts, http://dx.doi.org/10.1158/1055-9965.EPI-09-0762 [1], while exposure in mice increased platelet counts http://dx.doi.org/10.1371/journal.pone.0074974 [2]. Bone marrow megakaryocyte (MK) numbers were also increased in exposed mice, as determined qualitatively. The data presented here are from a quantitative evaluation of MK numbers in the bone marrow histopathological slides from the previous FA exposure experiments in mice. Bone marrow slides were prepared using a single 5 μm section of femur from 2 mice randomly selected from each exposure group (n=9) treated with 0, 0.5 and 3.0 mg/m3 FA by nose-only inhalation. MKs were systemically counted and average MK frequency was calculated as the total MK per slide divided by the number of fields evaluated. Data are presented visually as microscopy views and graphically as MK frequency

Evaluation of Drifting Snow Susceptibility Based on GIS and GA-BP Algorithms

Drifting snow, the flow of dispersed snow particles near ground level under the action of wind, is a major form of snow damage. When drifting snow occurs on railways, highways, and other transportation lines, it seriously affects their operational safety and results in drifting snow disasters. Drifting snow disasters frequently occur in the high latitudes of northwest China. At present, most scholars are committed to studying the prevention and control measures of drifting snow, but the prerequisite for prevention is to effectively evaluate the susceptibility of drifting snow along railways and highways to identify areas with a high risk of occurrence. Taking the Xinjiang Afukuzhun Railway as an example, this study uses a geographic information system (GIS) combined with on-site monitoring and surveys to establish a drifting snow susceptibility evaluation index system. The drifting snow susceptibility index (DSSI) is calculated through the weight of an evidence (WOE) model, and a genetic algorithm backpropagation (GA-BP) algorithm is used to obtain optimised evaluation index weights to improve the accuracy of model evaluation. The results show that the accuracies of the WOE model, WOE backpropagation (WOE-BP) model, and weight of evidence genetic algorithm backpropagation (WOE-GA-BP) model are 0.747, 0.748, and 0.785, respectively, indicating that the method can be effectively applied to evaluate drifting snow susceptibility

Bone marrow injury induced via oxidative stress in mice by inhalation exposure to formaldehyde.

OBJECTIVE: Formaldehyde, a ubiquitous environmental pollutant has been classified as a human leukemogen. However, toxicity of formaldehyde in bone marrow, the target site of leukemia induction, is still poorly understood. METHODOLOGY/PRINCIPAL FINDINGS: To investigate bone marrow toxicity (bone marrow pathology, hematotoxicity) and underlying mechanisms (oxidative stress, inflammation, apoptosis) in formaldehyde-exposed mice. Male Balb/c mice were exposed to formaldehyde (0, 0.5, and 3.0 mg/m(3)) by nose-only inhalation for 8 hours/day, over a two week period designed to simulate a factory work schedule, with an exposure-free "weekend" on days 6 and 7, and were sacrificed on the morning of day 13. Counts of white blood cells, red blood cells and lymphocytes were significantly (p<0.05) decreased at 0.5 mg/m(3) (43%, 7%, and 39%, respectively) and 3.0 mg/m(3) (52%, 27%, and 43%, respectively) formaldehyde exposure, while platelet counts were significantly increased by 109% (0.5 mg/m(3)) and 67% (3.0 mg/m(3)). Biomarkers of oxidative stress (reactive oxygen species, glutathione depletion, cytochrome P450 1A1 and glutathione s-transferase theta 1 expression), inflammation (nuclear factor kappa-B, tomour necrosis factor alpha, interleukin-1 beta), and apoptosis (activity of cysteine-aspartic acid protease 3) in bone marrow tissues were induced at one or both formaldehyde doses mentioned above. CONCLUSIONS/SIGNIFICANCE: Exposure of mice to formaldehyde by inhalation induced bone marrow toxicity, and that oxidative stress, inflammation and the consequential apoptosis jointly constitute potential mechanisms of such induced toxicity