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

Oral vaccination of juvenile carp (Cyprinus carpio) and gilthead seabream (Spams aurata) with bioencapsulated Vibrio anguillarum bacterin

In this study Artemia nauplii were used as an antigen delivery system for oral vaccination of young fish with a Vibrio anguillarum bacterin. Juvenile carp of 15-, 29- and 58-days old (stomachless fish) and gilthead seabream of 57- and 69-days old (stomach-containing fish) were fed with this bioencapsulated bacterin. Antigen transport in the endgut was studied using immunohistochemical techniques. In carp of 29- and 58-days old antigenic determinants of V. anguillarum were transported from lumen to supranuclear vacuoles in carp and finally to intraepithelial macrophages. In carp of 15-days old a similar transport was observed, only macrophages could not be found. In gilthead seabream a different (more diffuse) transport was observed and no macrophages were found. Ten weeks after oral vaccination fish of both species received an intramuscular booster. Three weeks later, serum was collected and measured for specific antibodies by ELISA. Carp orally vaccinated at 15- or 29-days old showed a significantly lower response after the booster compared with non-vaccinated controls. However, carp orally vaccinated at 58-days old tended to have a higher antibody response compared with the control. Seabream orally vaccinated at 57 or 69 days showed significantly higher secondary responses compared with the control. It is concluded that oral Vaccination with bioencapsulated vaccines is very promising when applied at the right age. Application at too young an age seems to induce immunosuppression, instead of immunisation

Characteristics of Anemia in Elderly: A Hospital Based Study in South India

Anemia is a common concern in older people and can have significant morbidity and mortality. Because anemia is a sign, not a diagnosis, an evaluation is almost always warranted to identify the underlying cause. The purpose of this study was to study the clinical profile of elderly patients with anemia and to study characteristics of hematological types of anemia in such patients as well as the closest possible etiological profile. Hundred patients above the age of 60 years were included in the study. Clinical profile with laboratory studies of Hemoglobin and diagnostic tests to fix the etiology. Majority of patients had normocytic blood picture. Renal failure was the most common underlying chronic disease. Significant number of patients were on non steroidal anti-inflammatory drugs which could contribute to the anaemia. 14% of the patients had an underlying malignancy. 73.3% of the patients in the microcytic group had an underlying GI lesion on endoscopy. Identifying anemia as an important aspect of a comprehensive geriatric assessment is absolutely essential further to clinical detection. Confirming the type of anemia is critical to direct the investigation for profiling the etiology since it is well known that the treatment of anemia goes a long way in improving the overall outcome and quality of life