Platform for establishing interlaboratory reproducibility of selected reaction monitoring-based mass spectrometry peptide assays.

Mass spectrometry (MS) is an attractive alternative to quantification of proteins by immunoassays, particularly for protein biomarkers of clinical relevance. Reliable quantification requires that the MS-based assays are robust, selective, and reproducible. Thus, the development of standardized protocols is essential to introduce MS into clinical research laboratories. The aim of this study was to establish a complete workflow for assessing the transferability and reproducibility of selected reaction monitoring (SRM) assays between clinical research laboratories. Four independent laboratories in North America, using identical triple-quadrupole mass spectrometers (Quantum Ultra, Thermo), were provided with standard protocols and instrumentation settings to analyze unknown samples and internal standards in a digested plasma matrix to quantify 51 peptides from 39 human proteins using a multiplexed SRM assay. The interlaboratory coefficient of variation (CV) was less than 10% for 25 of 39 peptides quantified (12 peptides were not quantified based upon hydrophobicity) and exhibited CVs less than 20% for the remaining peptides. In this report, we demonstrate that previously developed research platforms for SRM assays can be improved and optimized for deployment in clinical research environments.

Distribution, host range, and climatic constraints on Centistes gasseni (Hymenoptera: Braconidae), a South American parasitoid of cucumber beetles, Diabrotica spp. (Coleoptera: Chrysomelidae).

The genus Diabrotica includes a large number of pest species, including some of the most important crop pests of the Americas. The parasitoid Centistes gasseni Shaw is the first braconid to be described parasitizing Diabrotica in South America, and high natural infestations are reported. Field and experimental observations on the host range, distribution and biology of this parasitoid are described. Centistes gasseniwas collected in southern Brazil, eastern Paraguay and northeastern Argentina, in a region comprising humid lowlands and highlands, and cool temperate to warm subtropical climates, with regular rainfall in excess of 1300 mm. Three Diabroticaspecies, D. limitata (Sahlberg), D. speciosa (Germar) and D. viridula (Fabricius) were found to host the parasitoid, with mean percent parasitism of 5.4, 2.0 and 1.0%, respectively. Diabrotica speciosa and D. viridula are the two most important pest Diabroticaspecies in South America. Laboratory experiments with field-collected beetles and parasitoid cocoons indicated that C. gasseni overwinters in adult host beetles, remaining dormant in its live host below developmental temperatures. A potential distribution of C. gasseni in North America is proposed based on its known climatic range and the distribution of the main pest species of adult overwintering North American Diabrotica.

Effect of a phloxine B-cucurbitacin bait on diabroticite beetles (Coleoptera: Chrysomelidae).

Cucurbitacin E glycoside, extracted from a bitter mutant of Hawkesbury watermelon [Citrulls lanatus (Thunb.) Matsum. & Nakai (Syn. Citrullus vulgaris Schrad)] is the active ingredient of a feeding stimulant for the corn rootworm complex. It is the primary component of a water-soluble bait that can be combined with toxins for adult diabroticite control. Studies were conducted using phloxine B (D&C Red 28), a xanthene dye, as the toxin. This dye was efficacious against Diabrotica undecimpunctata howardi Barber, spotted cucumber beetle, and Acalymma vittatum (F.), striped cucumber beetle, in cucumber plots and could be recovered from cucumber leaves for 8 d after treatment. The average amount of dye recovered per dead spotted cucumber beetle at 8 d after treatment was 0.173 microg. Concentrated and sugar-free fermented forms of the watermelon extract were developed and compared with the fresh juice in field applications on cucumber plants. There was no significant difference in mortality of beetles from phloxine B-bait prepared with fresh, fermented, or concentrated extract, although in laboratory studies, fermented juice had higher feeding stimulant activity.