Fluorescence Imaging of Posterior Spiracles from Second and Third Instars of Forensically Important Chrysomya rufifacies (Diptera: Calliphoridae).

Entomological protocols for aging blowfly (Diptera: Calliphoridae) larvae to estimate the time of colonization (TOC) are commonly used to assist in death investigations. While the methodologies for analyzing fly larvae differ, most rely on light microscopy, genetic analysis, or, more rarely, electron microscopy. This pilot study sought to improve resolution of larval stage in the forensically important blowfly Chrysomya rufifacies using high-content fluorescence microscopy and biochemical measures of developmental marker proteins. We established fixation and mounting protocols, defined a set of measurable morphometric criteria and captured developmental transitions of 2nd instar to 3rd instar using both fluorescence microscopy and anti-ecdysone receptor Western blot analysis. The data show that these instars can be distinguished on the basis of robust, nonbleaching, autofluorescence of larval posterior spiracles. High-content imaging techniques using confocal microscopy, combined with morphometric and biochemical techniques, may therefore aid forensic entomologists in estimating TOC.

Contemporary Pacific and Western perspectives on `awa (Piper methysticum) toxicology.

In 2010, a National Science Foundation project in Hawai`i assembled a collaboration of Pacific indigenous scientists, Hawaiian cultural practitioners and scientists trained in Western pharmacology. The objective of the collaborative project was to study Kava, a culturally significant Pacific beverage, and to address and ultimately transcend, long-standing barriers to communication and collaboration between these groups. Kava is a product of the `awa plant (Piper methysticum) that has been used ceremonially and medicinally throughout the history of Pacific Island cultures, and is now in widespread recreational and nutraceutical use in the US. This project, culminating in 2015, has enriched the participants, led to published work that integrates cultural and Western pharmacologic perspectives and established a paradigm for collaboration. This review paper integrates cultural and Western perspectives on efficacy, toxicity and the future cultural and commercial significance of `awa in the Pacific. Here we present a detailed review of traditional and non-traditional kava usage, medicinal efficacy and potential toxicological concerns. Recent mechanistic data on physiological action and potential pathological reactions are evaluated and interpreted.