Investigating the effect of host plant identity on instar number in fall webworm, a common generalist herbivore.

For herbivorous insects with a broad diet breadth, host plant identity can influence larval development by either accelerating or delaying growth. For some species of Lepidoptera, the number of larval instars varies depending on the host plant's identity. Fall webworm (Hyphantria cunea, Drury) is a polyphagous herbivore that feeds on over 450 host plants worldwide. Of the 2 morphotypes (red- and black-head) of fall webworm, the number of instars for the red-head fall webworms has not been characterized. Given its broad diet breadth, fall webworm developmental stages may vary with plant identity. We investigated whether host plant identity affected the number of instars observed during red-head fall webworm development. We measured the head capsules of over 6,000 fall webworm larvae reared on 6 different plants commonly eaten by fall webworms in Colorado. We modeled head capsule widths as Gaussian mixture models, with a Gaussian distribution that corresponded to each instar. We show that our red-head fall webworms varied in number of instars depending on the identity of their host plant upon which they fed. We found that red-head fall webworm exhibited 7 instars on 5 of the host plants and 8 instars on 1 host plant that we studied. Our results for the number of instars for red-head fall webworm are consistent with reports of the number of instars for black-head fall webworm. Our research provides insight into the influence of host plant identity on fall webworm development, which can be used to advance lab and field research of this species.

Excerno: Using Mutational Signatures in Sequencing Data to Filter False Variants Caused by Clinical Archival.

The accurate detection of point mutations from pathology slides using sequencing data is of great importance in cancer genomics and precision oncology. Formalin-fixation paraffin-embedding (FFPE) is a widely used technique to preserve pathology tissues. The FFPE process introduces artificial C > T mutations in next-generation sequencing, so we set out to develop excerno, a method to score and filter such spurious variants. FFPE mutational artifacts follow a mutational signature. By using the FFPE signature and Bayes' formula, we can calculate the probability of a mutation resulting from the FFPE process and use this probability to filter FFPE variants. We implement this method as the excerno R package. We tested excerno by simulating mutations across all 60-baseline mutational signatures from the Catalog of Somatic Mutations in Cancer (COSMIC) and combining them with mutations following the FFPE mutational signature. The sensitivity and specificity of excerno are adversely affected by the cosine similarity between the baseline and FFPE signatures (cosFFPE). Higher percentages of FFPE mutations (pctFFPE) result in increased sensitivity and reduced specificity. The specificity and sensitivity of excerno can be predicted as linear model with an interaction term using cosFFPE and pctFFPE, with an R2of0.84 and 0.79, respectively. Finally, we tested excerno using six RNA sequencing cancer samples and observed concordant trends of specificity and sensitivity with respect to our simulated data. The excerno R package can be used to annotate and filter FFPE-induced mutations in cancer genomics. Our method is adversely affected by cosFFPE and pctFFPE.

A simple method for non-denaturing purification of biotin-tagged proteins through competitive elution with free biotin.

The use of avidin or streptavidin in the purification of biotinylated proteins has been highly restricted due to the harsh and denaturing elution conditions. Here, we use biotinylated bovine serum albumin as a working model to demonstrate a simple and rapid method for biotin-tagged protein purification under non-denaturing conditions. The biotinylated bovine serum albumin is specifically bound to the anti-biotin antibody agarose beads and competitively eluted with free biotin under near-neutral conditions. The optimized elution conditions include using 4 mg/ml biotin (pH 8.5) as the elution buffer and allowing the buffer to incubate with agarose beads for 30 min prior to elution. The elution recovery rate is over 85% without apparent protein denaturation. The method is applicable for both immunoprecipitation and column chromatography.

Delayed postoperative agitation in a child after preoperative midazolam.

We present a case report of a 5-year-old boy who had severe agitation after PACU discharge after midazolam premedication for minor otologic surgery. He was treated over several hours in the emergency department with several doses of midazolam and phenobarbital, without resolution of the agitation. Finally, a single dose of flumazenil (0.2 mg) effectively reversed the child's agitation. The incidence and potential mechanisms for postoperative agitation are discussed, with a focus on the potential for midazolam-induced behavioral reactions. This information has nursing implications for postoperative discharge instructions and follow-up.