Methods for detecting autophagy and determining autophagy-induced cell deathThis review is one of a selection of papers published in a Special Issue on Oxidative Stress in Health and Disease.

2010 ◽  
Vol 88 (3) ◽  
pp. 285-295 ◽  
Author(s):  
Yongqiang Chen ◽  
Meghan B. Azad ◽  
Spencer B. Gibson

Autophagy is an intracellular lysosomal degradation process, which in the case of macroautophagy, is characterized by the formation of double-membraned autophagosomes. Enhanced under stress conditions, autophagy can function to promote cell survival or cell death depending on the type of cellular stress. Interest in autophagy has increased substantially in the past several years as new research implicates this “self-eating” pathway in cell growth, development, and many human diseases. Various methods have been developed for detecting autophagy; however, the implementation of these methods and the interpretation of the results often vary between studies, and a more standardized approach is required. In this review, we summarize the current methods available for detecting autophagy and for determining its contribution to cell death. Furthermore, we discuss the critical points for the successful application of these methods based on experiences from our laboratory and from other research groups.

2006 ◽  
Vol 84 (1) ◽  
pp. 93-109 ◽  
Author(s):  
Michael P. Czubryt ◽  
Leon Espira ◽  
Lise Lamoureux ◽  
Bernard Abrenica

In the past decade, increasing attention has been paid to the importance of sex in the etiology of cardiac dysfunction. While focus has been primarily on how sex modulates atherogenesis, it is becoming clear that sex is both a predictor of outcome and an independent risk factor for a number of other cardiac diseases. Animal models and human studies have begun to shed light on the mechanisms by which sex influences the function of cardiomyocytes in health and disease. This review will survey the current literature on cardiac diseases that are influenced by sex and discuss the intracellular mechanisms by which steroid sex hormones affect heart function. A theory on how sex may regulate myocardial energy metabolism to affect disease susceptibility and progression will be presented, as well as a discussion of how sex may influence outcomes of experiments on isolated cardiomyocytes by epigenetic marking.


2012 ◽  
Vol 27 ◽  
pp. 463-496 ◽  
Author(s):  
Max Diem ◽  
Miloš Miljković ◽  
Benjamin Bird ◽  
Tatyana Chernenko ◽  
Jen Schubert ◽  
...  

This paper summarizes the progress achieved over the past fifteen years in applying vibrational (Raman and IR) spectroscopy to problems of medical diagnostics and cellular biology. During this time, a number of research groups have verified the enormous information content of vibrational spectra; in fact, genomic, proteomic, and metabolomic information can be deduced by decoding the observed vibrational spectra. This decoding process is aided enormously by the availability of high-power computer workstations and advanced algorithms for data analysis. Furthermore, commercial instrumentation for the fast collection of both Raman and infrared microspectral data has rendered practical the collection of images based solely on spectral data. The progress in the field has been manifested by a steady increase in the number and quality of publications submitted by established and new research groups in vibrational biological and biomedical arenas.


2009 ◽  
Vol 34 (3) ◽  
pp. 488-492 ◽  
Author(s):  
Terry E. Graham

While scientists have routinely measured muscle glycogen in many metabolic situations for over 4 decades, there is surprisingly little known regarding its regulation. In the past decade, considerable evidence has illustrated that the carbohydrate stores in muscle are not homogeneous, and it is very likely that metabolic pools exist or that each granule has independent regulation. The fundamental aspects appear to be associated with a complex set of proteins that associate with both the granule and each other in a dynamic fashion. Some of the proteins are enzymes and others play scaffolding roles. A number of the proteins can translocate, depending on the metabolic stimulus. These various processes appear to be the mechanisms that give the glycogen granule precise yet dynamic regulation. This may also allow the stores to serve as an important metabolic regulator of other metabolic events.


2006 ◽  
Vol 84 (4) ◽  
pp. 651-667 ◽  
Author(s):  
Christopher P. Trobacher ◽  
Adriano Senatore ◽  
John S. Greenwood

Cysteine proteinases are ubiquitously involved in programmed cell death (PCD) in multicellular organisms. In animals, one group of cysteine proteinases, the cysteine-dependent aspartate-specific proteinases (caspases), are involved in a proteolytic signalling cascade that controls apoptosis, the most studied form of PCD. The enzymes act as both masterminds and executioners in apoptotic cell death. In plants, members of the metacaspase family, as well as those of the papain-like and legumain families, of cysteine proteinases have all been implicated in PCD. These enzymes often belong to sizeable gene families, with Arabidopsis having 9 metacaspase, 32 papain-like, and 4 legumain genes. This redundancy has made it difficult to ascertain the functional importance of any particular enzyme in plant PCD, as many are often expressed in a given tissue undergoing PCD. As yet, mechanisms similar to the apoptotic caspase cascade in animals have not been uncovered in plants and, indeed, may not exist. Are the various cysteine proteinases, so often implicated in plant PCD, merely acting as minions in the process? This review will outline reports of cysteine proteinases associated with plant PCD, discuss problems in determining the function of specific proteases, and suggest avenues for determining how these enzymes might be regulated and how PCD pathways upstream of protease expression and activation might operate.


2017 ◽  
Vol 95 (10) ◽  
pp. 1163-1169 ◽  
Author(s):  
Adriana Adameova ◽  
Jaroslav Hrdlicka ◽  
Adrian Szobi ◽  
Veronika Farkasova ◽  
Katarina Kopaskova ◽  
...  

Long-lasting ischemia can result in cell loss; however, repeated episodes of brief ischemia increase the resistance of the heart against deleterious effects of subsequent prolonged ischemic insult and promote cell survival. Traditionally, it is believed that the supply of blood to the ischemic heart is associated with release of cytokines, activation of inflammatory response, and induction of necrotic cell death. In the past few years, this paradigm of passive necrosis as an uncontrolled cell death has been re-examined and the existence of a strictly regulated form of necrotic cell death, necroptosis, has been documented. This controlled cell death modality, resembling all morphological features of necrosis, has been investigated in different types of ischemia-associated heart injuries. The process of necroptosis has been found to be dependent on the activation of RIP1–RIP3–MLKL axis, which induces changes leading to the rupture of cell membrane. This pathway is activated by TNF-α, which has also been implicated in the cardioprotective signaling pathway of ischemic preconditioning. Thus, this review is intended to describe the TNF-α-mediated signaling leading to either cell survival or necroptotic cell death. In addition, some experimental data suggesting a link between heart dysfunction and the cellular loss due to necroptosis are discussed in various conditions of myocardial ischemia.


2006 ◽  
Vol 84 (6) ◽  
pp. 1045-1050 ◽  
Author(s):  
Ravindra Aurade ◽  
Senigala K. Jayalakshmi ◽  
Kuruba Sreeramulu

A P-glycoprotein-like protein (Ha-Pgp) was detected in a membrane preparation from the insecticide-resistant pest Helicoverpa armigera (Lepidoptera: Noctüidae) using C219 antibodies that are directed towards an epitope in the nucleotide-binding domains. This protein was partially purified and found to be a glycoprotein displaying ATPase activity. SDS–PAGE confirmed that a high molecular mass glycoprotein (150 kDa) was overexpressed in resistant pests, but was not detected in susceptible pests. The partially purified Ha-Pgp ATPase was reconstituted into proteoliposomes and it was found that some insecticides, namely, monocrotophos, endosulfan, cypermethrin, fenvalerate, and methylparathion, stimulated the ATPase activity. The effect of various inhibitors on partially purified Ha-Pgp showed that orthovanadate is a potent inhibitor of its ATPase activity, inhibiting it by 90% at a concentration of 2 mmol/L. Other inhibitors, such as EDTA, sodium azide, and molybdate resulted in only a 20% decrease in activity. Details of the structure and function of Ha-Pgp will be important in the development of strategies to overcome insecticide resistance in this pest.


2006 ◽  
Vol 84 (3-4) ◽  
pp. 499-507 ◽  
Author(s):  
Daniel Stoffler ◽  
Kyrill Schwarz-Herion ◽  
Ueli Aebi ◽  
Birthe Fahrenkrog

Small ions and molecules can traverse the nuclear pore complex (NPC) simply by diffusion, whereas larger proteins and RNAs require specific signals and factors that facilitate their passage through the NPC. Our understanding of the factors that participate and regulate nucleocytoplasmic transport has increased tremendously over the past years, whereas the actual translocation step through the NPC has remained largely unclear. Here, we present and discuss recent findings on the interaction between the NPC and transport receptors and provide new evidence that the NPC acts as a constrained diffusion pore for molecules and particles without retention signal and as an affinity gate for signal-bearing cargos.


2005 ◽  
Vol 39 (4) ◽  
pp. 293-296 ◽  
Author(s):  
Ian G. Anderson ◽  
Lucy A. Tedd

PurposeTo introduce the theme of this special issue which contains a selection of papers presented at the Association for History and Computing UK (AHC‐UK) annual conference in 2004.Design/methodology/approachThe role of the AHC in the UK is described and the reasons for choosing the theme of the 2004 conference, Recasting the Past: Digital Histories, are outlined along with the original call for papers.FindingsThe contributors to this issue come from a wide geographic area and reflect the delegates at the conference by being archivists, historians, librarians and researchers.Originality/valueProvides an introduction to the special issue.


2021 ◽  
Vol 12 (5) ◽  
Author(s):  
Chan Li ◽  
Zhaoya Liu ◽  
Qian Xu ◽  
Huihui Peng ◽  
Jing Cao ◽  
...  

AbstractAutophagy, a well-observed intracellular lysosomal degradation process, is particularly important to the cell viability in diabetic cardiomyopathy (DCM). Peroxidasin (PXDN) is a heme-containing peroxidase that augments oxidative stress and plays an essential role in cardiovascular diseases, while whether PXDN contributes to the pathogenesis of DCM remains unknown. Here we reported the suppression of cell viability and autophagic flux, as shown by autophagosomes accumulation and increased expression level of LC3-II and p62 in cultured H9C2 and human AC16 cells that treated with 400 μM palmitate acid (PA) for 24 h. Simultaneously, PXDN protein level increased. Moreover, cell death, autophagosomes accumulation as well as increased p62 expression were suppressed by PXDN silence. In addition, knockdown of PXDN reversed PA-induced downregulated forkhead box-1 (FoxO1) and reduced FoxO1 phosphorylation, whereas did not affect AKT phosphorylation. Not consistent with the effects of si-PXDN, double-silence of PXDN and FoxO1 significantly increased cell death, suppressed autophagic flux and declined the level of FoxO1 and PXDN, while the expression of LC3-II was unchanged under PA stimulation. Furthermore, inhibition of FoxO1 in PA-untreated cells induced cell death, inhibited autophagic flux, and inhibited FoxO1 and PXDN expression. Thus, we come to conclusion that PXDN plays a key role in PA-induced cell death by impairing autophagic flux through inhibiting FoxO1, and FoxO1 may also affect the expression of PXDN. These findings may develop better understanding of potential mechanisms regarding autophagy in insulin-resistant cardiomyocytes.


2017 ◽  
Vol 30 (2) ◽  
pp. 3-16 ◽  
Author(s):  
Sarah Marks

This article will briefly explore some of the ways in which the past has been used as a means to talk about psychotherapy as a practice and as a profession, its impact on individuals and society, and the ethical debates at stake. It will show how, despite the multiple and competing claims about psychotherapy’s history and its meanings, historians themselves have, to a large degree, not attended to the intellectual and cultural development of many therapeutic approaches. This absence has the potential consequence of implying that therapies have emerged as value-free techniques, outside of a social, economic and political context. The relative neglect of psychotherapy, by contrast with the attention historians have paid to other professions, particularly psychiatry, has also underplayed its societal impact. This article will foreground some of the instances where psychotherapy has become an object of emerging historical interest, including the new research that forms the substance of this special issue of History of the Human Sciences.


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