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Aims: To devise a protocol for heterologous expression and purification of a partial toxic portion of the Bacillus thuringiensis (Bt) vegetative insecticidal protein Vip3A and using it as an antigen for anti-Vip3A polyclonal antibody development. Also, to evaluate the regulation of Vip3A secretion into culture supernatants (SNs) of different Bt strains based on this antibody., Methods and Results: A primer pair was designed to amplify partially the toxic portion of the vip3A gene from the HD125 strain. The amplicon was cloned in expressing vector to produce a ~35 kDa peptide, which was HPLC-purified prior to rabbit immunizations. The serum containing the polyclonal anti-Vip3A antibody demonstrated a detection sensitivity of 0·4 ng mm -2 for the antigen in slot-blot experiments. Seven Bt strains from different origins were assessed regarding their temporal secretion of Vip3A toxin. ELISA results showed a strain-specific temporal regulation of Vip3A secretion in culture for the temperate isolates, with no detection of the toxin for the tropical strains, even when the presence of the gene was confirmed by PCR and sequencing., Conclusions: Conformational variation in the toxic portion of Vip3A may explain lack of its detection in the tropical strains. Isolates from the same subspecies display physiological variability in proteins' secretion into culture SNs, which can affect screening procedures for more effective strains/toxins., Significance and Impact of the Study: Immunoassays based on the developed anti-Vip3A antibody can be useful in a variety of basic studies. This method can be also coupled with toxicity assays on target insects, for more efficient screening methods of novel Bt strains/toxins with biocontrol applicability., (© 2018 The Society for Applied Microbiology.)

For determination of whether plasma 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (DDE) pesticide levels (< or =1-32 ppb) are associated with immune suppression or DNA damage in lymphocytes, 302 individuals residing in Moore County, North Carolina, in 1994-1996 provided a blood specimen, underwent a skin test, and answered a questionnaire concerning factors affecting plasma organochlorine pesticide levels and the immune system. The blood specimens were analyzed for levels of plasma DDE (a metabolite of 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane), numbers and types of blood cells, immunoglobulin levels, mitogen-induced lymphoproliferative activity, and lymphocyte micronuclei. When DDE levels were categorized as 1 or less, more than 1 to 2, more than 2 to 4.3, more than 4.3 to 7.6, and more than 7.6 ppb, individuals with higher plasma DDE levels had lowered mitogen-induced lymphoproliferative activity (concanavalin A, range: 74,218 dropping to 55,880 counts per minute, p = 0.03) and modestly increased total lymphocytes (range: 2.0-2.3 x 10(3)/microl, p = 0.05) and immunoglobulin A levels (range: 210-252 mg/dl, p = 0.04). There were no consistent differences in response to the skin tests by plasma DDE levels. Plasma DDE levels were not associated with a higher frequency of micronuclei. The authors conclude that relatively low levels of plasma DDE are associated with statistically significant changes in immune markers, although the magnitude of the effects are of uncertain clinical importance.

We examined the effects of methanol and temperature on the reactivity of monoclonal antibodies specific to the insecticide etofenprox. When the antigen-antibody reaction was done at 4 degrees C in 10% methanol, the sensitivity in the enzyme immunoassay with each antibody was more than 10-fold higher than that measured at 37 degrees C. Although in 10% methanol one of the antibodies reacted equally with both etofenprox and the carbonate-derivative of etofenprox, in 50% methanol the antibody reacted with etofenprox, but not with the derivative.

Two polyclonal antibodies and three monoclonal antibodies specific to the etofenprox insecticide were prepared from rabbits and mice, respectively. The monoclonal antibodies were more reactive with etofenprox than the polyclonal antibodies by C-ELISA. Monoclonal antibody cl 205-65 was found to be the most tolerant to methanol, most highly reactive and most specific to etofenprox among the antibodies tested.