International Centre for Genetic Engineering and Biotechnology

About: International Centre for Genetic Engineering and Biotechnology is a nonprofit organization based out in Trieste, Italy. It is known for research contribution in the topics: Gene & Plasmodium falciparum. The organization has 2489 authors who have published 4516 publications receiving 195297 citations.

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

Abstract: In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure flux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation, it is imperative to target by gene knockout or RNA interference more than one autophagy-related protein. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways implying that not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular assays, we hope to encourage technical innovation in the field.

TDP-43 Mutations in Familial and Sporadic Amyotrophic Lateral Sclerosis

Abstract: Amyotrophic lateral sclerosis (ALS) is a fatal motor neuron disorder characterized pathologically by ubiquitinated TAR DNA binding protein (TDP-43) inclusions. The function of TDP-43 in the nervous system is uncertain, and a mechanistic role in neurodegeneration remains speculative. We identified neighboring mutations in a highly conserved region of TARDBP in sporadic and familial ALS cases. TARDBPM337V segregated with disease within one kindred and a genome-wide scan confirmed that linkage was restricted to chromosome 1p36, which contains the TARDBP locus. Mutant forms of TDP-43 fragmented in vitro more readily than wild type and, in vivo, caused neural apoptosis and developmental delay in the chick embryo. Our evidence suggests a pathophysiological link between TDP-43 and ALS.

In vitro: Response of plant growth regulators and antimalformins on conidia germination of Fusarium mangiferae and incidence of mango malformation

Abstract: Mango malformation is the most important and threatening disease of recent times, primarily because of persistent lacuna in complete understanding of its nature. Diverse Fusarium spp, including F. mangiferae, were found to be associated with the disease. Here, F. mangiferae from mango cv Dashehri was morphologically characterized. Typically, oval-shaped microconidia without septum and crescent-shaped macroconidia with 3-septate were more often observed, whereas not a single chlamydospore was detected. The length and width of micro- and macro-conidia were 7.5, 55, 3.2, and 3.5, respectively. The plant growth regulators such as NAA, GA3, BAP and ethrel were found to induce in vitro germination of conidia of F. mangiferae after 12 h. In contrast, antimalformin silver nitrate (AgNO3) inhibits conidial germination in vitro and none of conidia was germinated beyond 500 ppm, however antimalformin glutathione was highly effective in stimulating conidial germination of F. mangiferae in vitro at > 1000 ppm after 24 h. We observed that the response of F. mangiferae to germinate the conidia in vitro under influence of plant growth regulators and antimalformins is not coincided with earlier findings of reduced disease incidence by exogenous application of these compounds. The present findings do not authenticate the involvement of F. mangiferae in the disease, however hormonal imbalance, most probably ethylene, might be responsible for deformed functional morphology of panicle. Further, a signal transduction mechanism of stress-stimulated ethylene imbalance causing physio-morphological changes in reproductive organs of mango flower and thereby failure of fertilization and fruit set, which needs to be investigated.