ABSTRACT
Cnidarian venom research has lagged behind other toxinological fields due to technical difficulties in recovery of the complex venom from the microscopic nematocysts. Here we report a newly developed rapid, repeatable and cost effective technique of venom preparation, using ethanol to induce nematocyst discharge and to recover venom contents in one step. Our model species was the Australian box jellyfish (Chironex fleckeri), which has a notable impact on public health. By utilizing scanning electron microscopy and light microscopy, we examined nematocyst external morphology before and after ethanol treatment and verified nematocyst discharge. Further, to investigate nematocyst content or "venom" recovery, we utilized both top-down and bottom-up transcriptomics-proteomics approaches and compared the proteome profile of this new ethanol recovery based method to a previously reported high activity and recovery protocol, based upon density purified intact cnidae and pressure induced disruption. In addition to recovering previously characterized box jellyfish toxins, including CfTX-A/B and CfTX-1, we recovered putative metalloproteases and novel expression of a small serine protease inhibitor. This study not only reveals a much more complex toxin profile of Australian box jellyfish venom but also suggests that ethanol extraction method could augment future cnidarian venom proteomics research efforts.
Subject(s)
Cnidarian Venoms/genetics , Cubozoa/chemistry , Serine Proteinase Inhibitors/genetics , Animals , Australia , Cnidarian Venoms/chemistry , Microscopy, Electron, Scanning , Nematocyst/metabolism , Proteome/chemistry , Proteomics , Serine Proteinase Inhibitors/chemistry , TranscriptomeABSTRACT
Prairie Ridge Addiction Treatment Services turned to SAMHSA's Treatment Improvement Protocol (TIP) 33, "Treatment for Stimulant Use Disorders," to try to establish more effective practices for their fast-growing population of methamphetamine-addicted clients. Six years later, Prairie Ridge's executive director says that adopting the TIP's client-based treatment philosophy has enhanced the program's accessibility and results, not only for stimulant-abusing clients, but others as well. In this article he recounts how the TIP contents meshed with Prairie Ridge's preexisting treatment philosophy and practices; what they adopted and what they adapted from the TIP and why; counselors' responses during the transition; and outstanding issues.
