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

TRANSDERMAL ESTRADIOL PLUS ORAL MEDROXYPROGESTERONE ACETATE REPLACEMENT THERAPY IN PRIMARY AMENORRHEIC ADOLESCENTS - CLINICAL, HORMONAL AND METABOLIC ASPECTS

Twelve primary amenorrheic adolescents were treated with transdermal estradiol 100 mu g (Estraderm TTS-100 (R)) twice weekly for 3 weeks, plus MPA 10 mg per os daily (Provera(R)) for the last 11 days, following an interval of 1 week. A basic examination and a re-examination at 6- and 12-month intervals were carried out for clinical evaluation, hormonal assays, lipid, carbohydrate and bone metabolism. No significant changes were recorded on the FSH, LH, 17 beta-estradiol and PRL serum levels. A significant decrease of TC values and atheromatic indices 1 (TC/HDL) and 2 (LDL/HDL) was seen (P < 0.05) at the end of treatment. On the other hand a significant increase in apolipoproteins A1 was found. A beneficial effect was also recorded on the bone mass. Finally, no significant side-effects were reported. It is concluded that this kind of hormone replacement therapy is efficient and safe for the treatment of amenorrheic adolescents due to gonadal dysgenesis

Auditory function in young patients with chronic renal failure

Debate on the relationship between renal insufficiency and hearing loss continues mainly due to the advanced age and the possible accelerated presbycusis of the patients that have been studied in surveys. Hearing acuity was studied in 46 children and adolescents suffering from renal insufficiency. Sensorineural hearing loss (mainly high-frequency) of unknown cause was found in 14 patients (30.4%). Hearing loss was not influenced by the various haematological, biochemical and clinical parameters (type of renal disease, blood pressure, history of ototoxic drugs administration). However, hearing loss seemed to be affected by the method of management of the renal insufficiency (more in the haemodialysis group than in the peritoneal dialysis group). There were no significant changes in audiometric findings before and after haemodialysis