Evaluation of Demodex mite viability using motility and scattered light intensity.

Demodex mites have been suggested to have a role in various cutaneous and ocular disorders pathogenesis, such as rosacea or blepharitis. Evaluation of potential treatments with anti-Demodex effects is difficult because the viability of living mites needs to be evaluated during their exposure to the agent being tested. Mite viability is currently based solely on their observed movement. However, this method of assessing viability has significant limitations as mites may be resting, immobile or paralysed at any given observation point giving the observer a false impression of the organism's death. To overcome this limitation we evaluated a new quantitative method of evaluating the viability of Demodex mites by using scattered light intensity (SLI). We demonstrated that when combined with observation of mite motility, SLI provided increased accuracy of the evaluation of viability of mites being studied. This new viability assay will help address the technical challenges of mite viability experiments. Accurate evaluation of mite viability will enhance mite biology research and allow for more accurate in vitro toxicity assays of proposed anti-mite agents.

Interrogation of gossypol therapy in glioblastoma implementing cell line and patient-derived tumour models.

BACKGROUND: Glioblastoma (GBM), being a highly vascularised and locally invasive tumour, is an attractive target for anti-angiogenic and anti-invasive therapies. The GBM/endothelial cell response to gossypol/temozolomide (TMZ) treatment was investigated with a particular aim to assess treatment effects on cancer hallmarks. METHODS: Cell viability, endothelial tube formation and GBM tumour cell invasion were variously assessed following combined treatment in vitro. The U87MG-luc2 subcutaneous xenograft model was used to investigate therapeutic response in vivo. Viable tumour response to treatment was interrogated using immunohistochemistry. Combined treatment protocols were also tested in primary GBM patient-derived cultures. RESULTS: An endothelial/GBM cell viability inhibitory effect, as well as an anti-angiogenic and anti-invasive response, to combined treatment have been demonstrated in vitro. A significantly greater anti-proliferative (P=0.020, P=0.030), anti-angiogenic (P=0.040, P<0.0001) and pro-apoptotic (P=0.0083, P=0.0149) response was observed when combined treatment was compared with single gossypol/TMZ treatment response, respectively. GBM cell line and patient-specific response to gossypol/TMZ treatment was observed. CONCLUSIONS: Our results indicate that response to a combined gossypol/TMZ treatment is related to inhibition of tumour-associated angiogenesis, invasion and proliferation and warrants further investigation as a novel targeted GBM treatment strategy.