Guidelines for the use and interpretation of assays for monitoring autophagy (4th edition)1.

In 2008, we published the first set of guidelines for standardizing research in autophagy. Since then, this topic has received increasing attention, and many scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Thus, it is important to formulate on a regular basis updated guidelines for monitoring autophagy in different organisms. Despite numerous reviews, there continues to be confusion regarding acceptable methods to evaluate autophagy, especially in multicellular eukaryotes. Here, we present a set of guidelines for investigators to select and interpret methods to examine autophagy and related processes, and for reviewers to provide realistic and reasonable critiques of reports that are focused on these processes. These guidelines are not meant to be a dogmatic set of rules, because the appropriateness of any assay largely depends on the question being asked and the system being used. Moreover, no individual assay is perfect for every situation, calling for the use of multiple techniques to properly monitor autophagy in each experimental setting. Finally, several core components of the autophagy machinery have been implicated in distinct autophagic processes (canonical and noncanonical autophagy), implying that genetic approaches to block autophagy should rely on targeting two or more autophagy-related genes that ideally participate in distinct steps of the pathway. Along similar lines, because multiple proteins involved in autophagy also regulate other cellular pathways including apoptosis, not all of them can be used as a specific marker for bona fide autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field

Comparison of the Efficacy, Safety and Tolerability of Simvastatin and Pravastatin for Hypercholesterolemia

The efficacy and safety profile of simvastatin and pravastatin across their most commonly mended dosage ranges were compared in a double-blind, parallel, multicenter study in 550 patients with primary hypercholesterolemia. The study consisted of a 6-week placebo period followed by 18 weeks of active treatment. Patients were randomized to 10 mg of simvastatin or pravastatin once in the evening; doses were titrated at 6-week intervals to a maximum of 40 mg/day ff the low lipoprotein (LDL) cholesterol remained greater-than-or-equal-to 130 mg/dl (3.4 mmol/liter). Baseline characteristics were similar in both groups. At the end of the study with simvastatin and pravastatin, respectively, 30 and 14% continued to take the 10 mg dose and 48 and 66% were titrated to the maximal dose. After 18 weeks of treatment with simvastatin and pravastatin the mean percent decreases from baseline were, respectively, for total plasma cholesterol 27 and 19% (p <0.01 between groups), for LDL cholesterol 38 and 26% (p <0.01 between groups), for very low density lipoprotein cholesterol 30 and 16% (p <0.01 between groups), and for triglycerides 18 and 14% (p <0.05 between groups). The mean percent increase from baseline in high-density lipoprotein cholesterol was 15% with simvastatin compared to 12% with pravastatin (p <0.05 between groups). The efficacy goal of LDL cholesterol <130 mg/dl was achieved in 65% of the patients treated with simvastatin versus 39% of those treated with pravastatin (p <0.001). There was no significant difference between groups in the frequency of drug-related adverse experiences. A standard sleep questionnaire given at baseline and after 6, 12 and 18 weeks of treatment showed no impairment of sleep with either drug, Simvastatin was significantly more effective than pravastatin in reducing total, LDL and very low density lipoprotein cholesterol and triglycerides and in increasing high-density lipoprotein cholesterol, and had a comparable safety and tolerability profile