Re-thinking the Etiological Framework of Neurodegeneration

Neurodegenerative diseases are among the leading causes of disability and death worldwide. The disease-related socioeconomic burden is expected to increase with the steadily increasing life expectancy. In spite of decades of clinical and basic research, most strategies designed to manage degenerative brain diseases are palliative. This is not surprising as neurodegeneration progresses "silently" for decades before symptoms are noticed. Importantly, conceptual models with heuristic value used to study neurodegeneration have been constructed retrospectively, based on signs and symptoms already present in affected patients;a circumstance that may confound causes and consequences. Hence, innovative, paradigm-shifting views of the etiology of these diseases are necessary to enable their timely prevention and treatment. Here, we outline four alternative views, not mutually exclusive, on different etiological paths toward neurodegeneration. First, we propose neurodegeneration as being a secondary outcome of a primary cardiovascular cause with vascular pathology disrupting the vital homeostatic interactions between the vasculature and the brain, resulting in cognitive impairment, dementia, and cerebrovascular events such as stroke. Second, we suggest that the persistence of senescent cells in neuronal circuits may favor, together with systemic metabolic diseases, neurodegeneration to occur. Third, we argue that neurodegeneration may start in response to altered body and brain trophic interactions established via the hardwire that connects peripheral targets with central neuronal structures or by means of extracellular vesicle (E\-mediated communication. Lastly, we elaborate on how lifespan body dysbiosis may be linked to the origin of neurodegeneration. We highlight the existence of bacterial products that modulate the gut-brain axis causing neuroinflammation and neuronal dysfunction. As a concluding section, we end by recommending research avenues to investigate these etiological paths in the future. We think that this requires an integrated, interdisciplinary conceptual research approach based on the investigation of the multimodal aspects of physiology and pathophysiology. It involves utilizing proper conceptual models, experimental animal units, and identifying currently unused opportunities derived from human data. Overall, the proposed etiological paths and experimental recommendations will be important guidelines for future cross-discipline research to overcome the translational roadblock and to develop causative treatments for neurodegenerative diseases

Generation and characterization of a Leishmania tarentolae strain for site-directed in vivo biotinylation of recombinant proteins

Leishmania tarentolae is a non-human-pathogenic Leishmania species of growing interest in biotechnology, as it is well-suited for the expression of human recombinant proteins. For many applications it is desirable to express recombinant proteins with a tag allowing easy purification and detection. Hence, we adopted a scheme to express recombinant proteins with a His-tag and, additionally, to site-specifically in vivo biotinylate them for detection. Biotinylation is a relatively rare modification of endogenous proteins that allows easy detection with negligible cross-reactivity. Here, we established a genetically engineered L. tarentolae strain constitutively expressing the codon-optimized biotin-protein ligase from Escherichia coli (BirA). We thoroughly analyzed the strain for functionality using 2-D polyacrylamide-gel electrophoresis (PAGE), mass spectrometry, and transmission electron microscopy (TEM). We could demonstrate that neither metabolic changes (growth rate) nor structural abnormalities (TEM) occurred. To our knowledge, we show the first 2-D PAGE analyses of L. tarentolae. Our results demonstrate the great benefit of the established L. tarentolae in vivo biotinylation strain for production of dual-tagged recombinant proteins. Additionally, 2-D PAGE and TEM results give insights into the biology of L. tarentolae, helping to better understand Leishmania species. Finally, we envisage that the system is transferable to human-pathogenic species

Nuclear Receptor Research: Contributions from Latin America

The fact that virtually every single aspect of cell function and the complex network of regulatory processes in an organism involve crucial roles by nuclear receptors is not surprising at the present time. Accordingly, thousands of studies are published every year in hundreds of journals with very broad spectra, making it difficult for researchers in the field to find specific information of interest to them. This is the reason why Nuclear Receptor Research was born; to gather in one journal most of the advances covering all facets of this cardinal group of regulatory transcription factors.Fil: Napimoga, Marcelo Henrique. São Leopoldo Mandic Institute and Research Center. Laboratory of Immunology and Molecular Biology; BrasilFil: Galigniana, Mario Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental (i); ArgentinaFil: Migliorini Figueira, Ana Carolina. Brazilian Center for Research in Energy and Materials. Brazilian Biosciences National Laboratory; BrasilFil: Onate, Sergio A. . Universidad de Concepción; ChileFil: Castro Obregon, Susana. Universidad Nacional Autónoma de México. Instituto de Fisiología Celular; Méxic

Effective Design of Multifunctional Peptides by Combining Compatible Functions.

Multifunctionality is a common trait of many natural proteins and peptides, yet the rules to generate such multifunctionality remain unclear. We propose that the rules defining some protein/peptide functions are compatible. To explore this hypothesis, we trained a computational method to predict cell-penetrating peptides at the sequence level and learned that antimicrobial peptides and DNA-binding proteins are compatible with the rules of our predictor. Based on this finding, we expected that designing peptides for CPP activity may render AMP and DNA-binding activities. To test this prediction, we designed peptides that embedded two independent functional domains (nuclear localization and yeast pheromone activity), linked by optimizing their composition to fit the rules characterizing cell-penetrating peptides. These peptides presented effective cell penetration, DNA-binding, pheromone and antimicrobial activities, thus confirming the effectiveness of our computational approach to design multifunctional peptides with potential therapeutic uses. Our computational implementation is available at http://bis.ifc.unam.mx/en/software/dcf

Minimal inhibitory concentrations for the tested peptides.

<p>Minimal inhibitory concentrations for the tested peptides.</p

Identification and design of CPP-like sequences.

<p>(A) Methodology used to identify and design CPP-like sequences. First, a random forest predictor is trained on a set of known CPP and non-CPP sequences. The resulting probabilistic model is then used to identify CPP-like sequences and converted into an energy measure (E = 1 –P_CPP(S)) for novel peptide design by simulated-annealing optimization (see <a href="http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1004786#sec008" target="_blank">Materials and Methods</a>). (B) Algorithm performance during the design of CPP-like sequences. 32 random peptide sequences of length 16 were optimized with varying fragment lengths for 1000 iterations. Short fragments of more than 4 amino acids proved sufficient to transform non-CPP sequences to CPP-like sequences.</p

Antibacterial activity.

<p>DH5α <i>E</i>. <i>coli</i> cells were treated with varying concentrations of peptides ranging from 0 to 16 μM for the peptides and 0 to 48 μM for Ampicillin. The relative area under the growth curve measured the growth inhibition. Measurements from four repetitions distributed onto two different plates are shown as dots colored by their respective concentrations. Linear regressions are shown as blue lines and their respective standard errors as blue shades. Dashed lines indicate the mean area under the curve for the H2O controls and its 50% value respectively. Minimal inhibitory concentrations to achieve 50% or 100% of cell growth inhibition are reported.</p