The Enigmatic Role of C9ORF72 in Autophagy

Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disease characterized by the loss of motor neurons resulting in a progressive and irreversible muscular paralysis. Advances in large-scale genetics and genomics have revealed intronic hexanucleotide repeat expansions in the gene encoding C9ORF72 as a main genetic cause of ALS and frontotemporal dementia (FTD), the second most common cause of early-onset dementia after Alzheimer's disease. Novel insights regarding the underlying pathogenic mechanisms of C9ORF72 seem to suggest a synergy of loss and gain of toxic function during disease. C9ORF72, thus far, has been found to be involved in homeostatic cellular pathways, such as actin dynamics, regulation of membrane trafficking, and macroautophagy. All these pathways have been found compromised in the pathogenesis of ALS. In this review, we aim to summarize recent findings on the function of C9ORF72, particularly in the macroautophagy pathway, hinting at a requirement to maintain the fine balance of macroautophagy to prevent neurodegeneration

Proteomic and network analysis characterize stage-specific metabolism in Trypanosoma cruzi

<p>Abstract</p> <p>Background</p> <p><it>Trypanosoma cruzi </it>is a Kinetoplastid parasite of humans and is the cause of Chagas disease, a potentially lethal condition affecting the cardiovascular, gastrointestinal, and nervous systems of the human host. Constraint-based modeling has emerged in the last decade as a useful approach to integrating genomic and other high-throughput data sets with more traditional, experimental data acquired through decades of research and published in the literature.</p> <p>Results</p> <p>We present a validated, constraint-based model of the core metabolism of <it>Trypanosoma cruzi </it>strain CL Brener. The model includes four compartments (extracellular space, cytosol, mitochondrion, glycosome), 51 transport reactions, and 93 metabolic reactions covering carbohydrate, amino acid, and energy metabolism. In addition, we make use of several replicate high-throughput proteomic data sets to specifically examine metabolism of the morphological form of <it>T. cruzi </it>in the insect gut (epimastigote stage).</p> <p>Conclusion</p> <p>This work demonstrates the utility of constraint-based models for integrating various sources of data (e.g., genomics, primary biochemical literature, proteomics) to generate testable hypotheses. This model represents an approach for the systematic study of <it>T. cruzi </it>metabolism under a wide range of conditions and perturbations, and should eventually aid in the identification of urgently needed novel chemotherapeutic targets.</p

Cryptosporidium hominis Infection of the Human Respiratory Tract

Cryptosporidium oocysts, observed in a natural sputum sample of a patient with HIV, were further studied by using DNA markers to determine the species of the parasite. C. hominis was identified as the species infecting the patient’s respiratory tract, a finding that strengthens evidence regarding this pathogen’s role in human disease

Axonal Degeneration during Aging and Its Functional Role in Neurodegenerative Disorders

Aging constitutes the main risk factor for the development of neurodegenerative diseases. This represents a major health issue worldwide that is only expected to escalate due to the ever-increasing life expectancy of the population. Interestingly, axonal degeneration, which occurs at early stages of neurodegenerative disorders (ND) such as Alzheimer's disease, Amyotrophic lateral sclerosis, and Parkinson's disease, also takes place as a consequence of normal aging. Moreover, the alteration of several cellular processes such as proteostasis, response to cellular stress and mitochondrial homeostasis, which have been described to occur in the aging brain, can also contribute to axonal pathology. Compelling evidence indicate that the degeneration of axons precedes clinical symptoms in NDs and occurs before cell body loss, constituting an early event in the pathological process and providing a potential therapeutic target to treat neurodegeneration before neuronal cell death. Although, normal aging and the development of neurodegeneration are two processes that are closely linked, the molecular basis of the switch that triggers the transition from healthy aging to neurodegeneration remains unrevealed. In this review we discuss the potential role of axonal degeneration in this transition and provide a detailed overview of the literature and current advances in the molecular understanding of the cellular changes that occur during aging that promote axonal degeneration and then discuss this in the context of ND

Expression of CApy in oocysts and sporozoites.

<p>CApy is expressed in oocysts and sporozoites. Aliquots of NP-40 soluble (S) or insoluble (I) <i>C. parvum</i> oocyst and sporozoite fractions were resolved by 12% SDS-PAGE, transferred to nitrocellulose and probed with mouse anti-rCApy serum as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0031030#s2" target="_blank">Materials and Methods</a>.</p

Glycosylation of CApy in sporozoites.

<p><i>C. parvum</i> sporozoites were treated as recommended in the Enzymatic Carbo Release Kit™ (QAbio, San Mateo, CA) for identification of glycosylation. In brief, sporozoites were suspended in Carbo Release buffer, and denaturation buffer was added. After incubation at 100°C for 5 min, samples were chilled on ice and Triton-X was added. The enzymes PNGase, Sialidase, ß-Galactosidase, Glucosaminidase, O-Glycosidase were added alone or in combinations. Following incubation at 37°C for 16 hours, protein loading buffer was added and samples were incubated again at 100°C for 5 min. Proteins were separated by 12% SDS-PAGE, transferred to nitrocellulose and probed with mouse anti-rCApy serum.</p