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

Prise en charge de l'accident vasculaire cérébral (AVC) (phase préhospitalière et hospitalière), évaluation des facteurs de non admission directe en Unité Neuro-Vasculaire (UNV) de Bretonneau et recueil des connaissances de la filière par les médecins généralistes en Indre-Et-Loire (37)

TOURS-BU Médecine (372612103) / SudocSudocFranceF

High Frequency of Previous Abuse and Missed Diagnoses Prior to Abusive Head Trauma: A Consecutive Case Series of 100 Forensic Examinations

International audienceThis study describes the frequency of signs and symptoms of abuse and missed diagnoses prior to the diagnosis of abusive head trauma (AHT) in infants. Data were from a retrospective observational study of 100 consecutive cases of infants diagnosed with AHT over a seven-year period. The most frequent symptom leading to the diagnosis was a loss of consciousness (68%) that always occurred inside a home (parents' or nanny's), never outside. Diagnosis was established using criteria based on the child's lesions and the alleged history. Lesions leading to diagnosis were described: 99 per cent had multifocal subdural haematoma (SDH) located in four areas including lateral space, interhemispheric, tentorium cerebelli and vertex; 60 per cent had a rupture of bridging veins. Previous abuse was found in 79 per cent of cases, of whom 75 per cent underwent medical consultations that did not result in a diagnosis of abuse. The main signs and symptoms of previous abuse were repeated vomiting without fever or diarrhoea (62%), abnormal head circumference increase (49%) and bruises (38%). These results suggest a higher frequency of repeated abuse than previous studies and highlight the great challenges most professionals encounter to evoke and set the diagnosis of abuse. 'This study describes the frequency of signs and symptoms of abuse and missed diagnoses prior to the diagnosis of abusive head trauma (AHT) in infants' Key Practitioner MessagesPrior signs and symptoms suggestive of abuse are particularly frequent in infants diagnosed with AHT. Abuse must be detected as early as possible in order to avoid recurrences. Diagnostic criteria, based on clinical history and lesions including multifocal subdural haematoma, rupture of bridging veins, retinal haemorrhages, and spinal cord injury have been defined to enable diagnosis of AHT

The separation of alpha-2 macroglobulin into five components with differing electrophoretic and enzyme-binding properties

The alpha-2 macroglobulins from human serum and plasma were isolated by Bio-Gel P-300 and A5m gel filtration. The material showed a single peak on sedimentation velocity ultracentrifugation, a mol wt of 650,000 by sedimentation equilibrium ultracentrifugation, and a major precipitin arc in the alpha-2 macroglobulin region by immunoelectrophoresis against whole human serum. Two bands were observed in the alpha-2 macroglobulin region when acrylamide gel electrophoresis was performed with a pH 8.9 running gel. When a pH 7.8 gel was used, five electrophoretic species were observed. In both cases, the preaddition of stoichiometric amounts of trypsin or chymotrypsin added to alpha-2 macroglobulin resulted in disappearance of slower bands leaving only one band on acrylamide gel electrophoresis patterns. Preparative acrylamide gel electrophoresis separated alpha-2 macroglobulin obtained from Bio-Gel into five closely-spaced species. Separation was sufficiently adequate to show that those species of alpha-2 macroglobulin which bound trypsin and chymotrypsin were represented by slower moving species and that the fastest moving material had lost virtually all of the ability to bind these enzymes. Preparative acrylamide gel electrophoresis of a mixture of alpha-2 macroglobulin-trypsin complex and alpha-2 macroglobulin revealed that the fast moving component was alpha-2 macroglobulin-trypsin complex and that the slower moving material was unbound alpha-2 macroglobulin. The naturally occurring amidase activity of the alpha-2 macroglobulin using benzoylarginine-p-nitroanilide (BAPNA) as substrate was investigated and unlike its trypsin-binding activity, amidase activity was found to be of the same specific activity in all electrophoretic fractions. Binding of trypsin and chymotrypsin to alpha-2 macroglobulin revealed that alpha-2 macroglobulin maximally bound 2 moles of trypsin and 1 mole of chymotrypsin. When the enzymes were added simultaneously there was competition. Chymotrypsin added to alpha-2 macroglobulin before the addition of trypsin prevented all trypsin binding even though only one site was filled with chymotrypsin. These results were explained by the acrylamide gels which showed that 1 mole of chymotrypsin was sufficient to convert all the alpha-2 macroglobulin to a species with the fastest mobility which no longer binds additional enzyme

A new polychelidan lobster preserved with its eggs in a 165 Ma nodule

International audienc