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Obtaining the specific development time of each species of forensic interest is crucial for the estimation of an accurate and reliable Minimum Postmortem Interval (PMImin). In Argentina, Lucilia ochricornis (Wiedemann) and Lucilia purpurascens (Walker) (Diptera: Calliphoridae) were masked under the name Lucilia cluvia (Walker) for a long time still in forensic expertise. For this reason, the objective of this work is to deepen the study of the development time of these species and utilize this relevant information in the generation of different associated methods that can be used in forensics to estimate the PMI. Immature stages of L. ochricornis and L. purpurascens were reared in a brood chamber according to the following temperature treatments: 13.4, 15.1, 22.3, and 23.6°C. The development time of each stage/state of these flies was recorded as well as the resulting accumulated degree-hours (ADH), to build isomorphen diagrams and thermal summation models for each species. The development time and ADH were different between both species and their development stages. On the other hand, the methods provided for estimating PMImin provide the forensic entomologist more tools to reach accurate and reliable estimates., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The objective of this work was to evaluate the effect of freeze-dried pulps of Eugenia uniflora L. and Opuntia ficus - indica fruits on oxidative stability of n-3-enriched beef patties. Besides, the effect of freeze-dried pulps on cooking yield, color, texture parameters and sensory acceptance of patties was also evaluated. Eugenia uniflora L. (orange-skinned variety) and Opuntia ficus - indica (orange-skinned and red-skinned varieties) fruits were collected, their pulp was lyophilized and incorporated, as a natural source of antioxidant, in beef patties enriched with canola oil as a source of polyunsaturated fatty acids. Oxidation stability of samples was monitored by measuring thiobarbituric acid-reactive substances during refrigerated storage. Results show that all freeze-dried pulps limited lipid oxidation to an acceptable level during 15 days refrigerated storage, having Opuntia ficus - indica (red-skinned variety) the highest inhibitory activity. Besides, the addition of freeze-dried pulps increased the cooking yield and decreased the hardness of beef patties. Respect to sensorial parameters, patties elaborated with no added freeze-dried fruit pulp and patties elaborated with freeze-dried pulp of Opuntia ficus - indica (red-skinned variety) presented the highest overall acceptance of consumers, remaining as a future task to improve the color and taste of patties elaborated with freeze-dried pulps of Eugenia uniflora L. and Opuntia ficus - indica (orange-skinned variety). This research reveals the effectiveness of studied fruits to reduce lipid oxidation in beef patties. In addition, preliminary information about which sensorial parameters of these products should be improved in futures incorporation of pulps in the manufacture of healthy meat products., (© Association of Food Scientists & Technologists (India) 2020.)

The circadian clock at the hypothalamic suprachiasmatic nucleus (SCN) entrains output rhythms to 24-h light cycles. To entrain by phase-advances, light signaling at the end of subjective night (circadian time 18, CT18) requires free radical nitric oxide (NO•) binding to soluble guanylate cyclase (sGC) heme group, activating the cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG). Phase-delays at CT14 seem to be independent of NO•, whose redox-related species were yet to be investigated. Here, the one-electron reduction of NO• nitroxyl was pharmacologically delivered by Angeli's salt (AS) donor to assess its modulation on phase-resetting of locomotor rhythms in hamsters. Intracerebroventricular AS generated nitroxyl at the SCN, promoting phase-delays at CT14, but potentiated light-induced phase-advances at CT18. Glutathione/glutathione disulfide (GSH/GSSG) couple measured in SCN homogenates showed higher values at CT14 (i.e., more reduced) than at CT18 (oxidized). In addition, administration of antioxidants N-acetylcysteine (NAC) and GSH induced delays per se at CT14 but did not affect light-induced advances at CT18. Thus, the relative of NO• nitroxyl generates phase-delays in a reductive SCN environment, while an oxidative favors photic-advances. These data suggest that circadian phase-locking mechanisms should include redox SCN environment, generating relatives of NO•, as well as coupling with the molecular oscillator.

Chagas disease is a serious parasitic infection caused by Trypanosoma cruzi. Unfortunately, the current chemotherapeutic tools are not enough to combat the infection. The aim of this study was to evaluate the trypanocidal activity of benznidazole-loaded microparticles during the acute phase of Chagas infection in an experimental murine model. Microparticles were prepared by spray-drying using copolymers derived from esters of acrylic and methacrylic acids as carriers. Dissolution efficiency of the formulations was up to 3.80-fold greater than that of raw benznidazole. Stability assay showed no significant difference (P > 0.05) in the loading capacity of microparticles for 3 years. Cell cultures showed no visible morphological changes or destabilization of the cell membrane nor haemolysis was observed in defibrinated human blood after microparticles treatment. Mice with acute lethal infection survived 100% after 30 days of treatment with benznidazole microparticles (50 mg kg-1 day-1). Furthermore, no detectable parasite load measured by quantitative polymerase chain reaction and lower levels of T. cruzi-specific antibodies by enzyme-linked immunosorbent assay were found in those mice. A significant decrease in the inflammation of heart tissue after treatment with these microparticles was observed, in comparison with the inflammatory damage observed in both infected mice treated with raw benznidazole and untreated infected mice. Therefore, these polymeric formulations are an attractive approach to treat Chagas disease.

The early arrival and colonization of species belonging to the family Calliphoridae (Insecta: Diptera) on a corpse represent one of the most reliable means of estimating minimum postmortem interval (PMImin). However, information on the development and life cycles of some Argentine species in this family is not complete. The objective of this work was to contribute new information regarding the larval body size of neotropical species that allow, through the construction of forensic methods, the estimation of a more precise and specific PMImin. This work was conducted on laboratory cultures of larvae of Lucilia ochricornis (Wiedemann) and Lucilia purpurascens (Walker) using as average temperatures: 13.4, 15.1, 22.6, and 23.3°C, which represent the four seasons of the year for the province of Salta. With this information, we constructed isomegalen diagrams and growth models for the obtained variables of larval length and body weight. The mean values of length and body weight differ between both species, indicating that L. purpurascens exceeded L. ochricornis in both variables. In contrast, within each species the mean length and weight remained unchanged between culture temperatures for the three larval instars. Isomegalen diagrams can be used for the entire range of temperatures worked in the laboratory, but the body size entered is approximate. The growth models allow the use of point data but are specific for each culture temperature used., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Introduction: Resective-reconstructive treatment of an abnormally invasive placenta, also known as conservative surgical management, allows a comprehensive treatment of the pathology in only one surgery; however, this alternative is not generally included in international consensus, as it requires specific training. Here, we report our experience of this type of treatment and its plausibility after training facilitated by interinstitutional collaboration via telemedicine. Materials and methods: A total of 48 women who were diagnosed with abnormally invasive placenta, before and after changes due to the resection-reconstruction protocol were included in the study. Results: In total, 14 conservative reconstructive procedures were performed with outcomes of a lower rate of bleeding, reduced transfusions and complications, and a shorter duration of hospitalization than women with hysterectomy. Conclusion: Conservative surgical management is a safe alternative when implemented at specialized centers by trained groups of professionals. Interinstitutional collaboration, using appropriate telemedicine is a safe and effective alternative to enable training in resective-conservative management of abnormally invasive placenta.

Aromatic nitro compounds are key building blocks for many industrial syntheses and are also components of explosives, drugs and pesticides. Due to the environmentally unfriendly experimental conditions involved in their chemical syntheses, industrial processes would benefit from the use of biocatalysts. Among potentially useful enzymes, N-oxygenases, whose role is to oxygenate primary amines, are becoming relevant. These enzymes are involved in different secondary metabolic pathways in Streptomyces and in few other bacteria, forming part of the enzyme pools implicated in antibiotic synthesis. In this work, a group of Streptomyces strains, whose biomass was obtained from simple and novel culture media, were identified as new sources of N-oxygenase activity. Furthermore, the use of unspecific metabolic stimulation strategies allowed substantial improvements in the activity of whole cells as biocatalysts. It is remarkable the 6 to 50-fold increase in nitro compound yields compared to the biotransformation under standard conditions when Streptomyces griseus was the biocatalyst. In addition, biocatalyst substrate acceptance was studied in order to determine the biocatalytic potential of this enzyme., (Copyright © 2021 Elsevier B.V. All rights reserved.)

It is well known that the splitting of tablets can bring serious risks to the health of the treated animals, e.g., the possible adverse reactions caused by overdoses of fenbendazole or aspirin. In this regard, this work aimed to evaluate, for the first time, the splitting behavior of commercial veterinary tablets and identifying the technological aspects that interfere in this process. Tablets were cut in halves using a tablet splitter and were analyzed regarding mass variation, mass loss, friability, and hardness. Microstructural and morphological evaluations were also performed. For most of the tablets, organic flavor additives provided more uniformity and cohesive matrix, which preserved its hardness after the cut and led to subdivision results within acceptable limits for mass measurements and friability. Apart from the microstructure, the most critical technological aspect for a correct splitting performance in such tablets was the presence of a score. Thus, the results presented here allow us to guide the manufacturing of veterinary drug products in order to produce tablets more adapted to the splitting process.

The main issue addressed in this review is the need for innovation in the artisanal production of dry fermented sausages-leveraging rather than discarding tradition, together with some practical strategies available to achieve it. Throughout the text, emphasis is placed on the autochthonous microbiota responsible for the identity and unique sensory characteristics of these products. The available strategies to introduce innovation in this manufacturing process rely on metabolic flexibility of microbial strains. In this sense, this review evaluates the application of several tools aimed at improving the quality and safety of artisanal dry fermented sausages focusing on the microbial community role. The most studied alternatives to enhance dry sausage production comprise the use of autochthonous starter cultures-including functional and/or probiotic strains, the production of bacteriocins, and the generation of bioactive peptides, which have been thoroughly covered herein. The purpose of this work is to review recent research about novel different strategies available for food technologists to improve safety and quality in the manufacture of dry fermented sausages. Additional support strategies-quality product registers and innovation through tradition-have been suggested as complementary actions towards a successful introduction of indigenous microbial communities into traditional dry sausage production., Competing Interests: The authors declare that they have no conflicts of interest regarding the publication of this paper., (Copyright © 2021 Noelia Zulema Palavecino Prpich et al.)

Antibodies are the most used technological tool in histochemistry. However, even with monoclonal antibodies, their standardization is difficult due to variation of biological systems as well as to variability due to the affinity and amplification of the signal arising from secondary peroxidase detection systems. In this article we combined two synthetic molecules to facilitate the standardization of a detection protocol of protein markers in histological sections. The first molecule was an aptamer, a 50-base single-stranded DNA fragment, which recognizes a PTEN tumor suppressor. The second molecule used was also another single stranded 18-base aptamer DNA fragment, which forms a quadruplex structure guanine box. This G-quadruplex recognizes and attaches a molecule of hemin, increasing the catalytic capacity for the hydrogen peroxide. Our results show how the correct structural design of DNA combining an aptamer together with the peroxidase-like DNAzyme allows to detect proteins in histological sections. This tool offers the standardization of the detection of prognostic markers in cancer, in quality and quantity, due to its synthetic nature and its 1:1 antigen:enzyme ratio. This is the first time that reproducible results have been presented in histological sections staining a cancer marker using a single-stranded DNA molecule with dual function.

Poly(itaconic acid) (PIA) was synthesized via conventional radical polymerization. Then, functionalization of PIA was carried out by an esterification reaction with the heterocyclic groups of 1,3-thiazole and posterior quaternization by N-alkylation reaction with iodomethane. The modifications were confirmed by Fourier transform infrared (FTIR) and proton nuclear magnetic resonance ( 1 H-NMR), as well as ζ-potential measurements. Their antimicrobial activity was tested against different Gram-negative and Gram-positive bacteria. After characterization, the resulting polymers were incorporated into gelatin with oxidized starch and glycerol as film adjuvants, and dopamine as crosslinking agent, to develop antimicrobial-active films. The addition of quaternized polymers not only improved the mechanical properties of gelatin formulations, but also decreased the solution absorption capacity during the swelling process. However, the incorporation of synthesized polymers increased the deformation at break values and the water vapor permeability of films. The antioxidant capacity of films was confirmed by radical scavenging ability and, additionally, those films exhibited antimicrobial activity. Therefore, these films can be considered as good candidates for active packaging, ensuring a constant concentration of the active compound on the surface of the food, increasing products' shelf-life and reducing the environmental impact generated by plastics of petrochemical origin.

The RepeatsDB database (URL: https://repeatsdb.org/) provides annotations and classification for protein tandem repeat structures from the Protein Data Bank (PDB). Protein tandem repeats are ubiquitous in all branches of the tree of life. The accumulation of solved repeat structures provides new possibilities for classification and detection, but also increasing the need for annotation. Here we present RepeatsDB 3.0, which addresses these challenges and presents an extended classification scheme. The major conceptual change compared to the previous version is the hierarchical classification combining top levels based solely on structural similarity (Class > Topology > Fold) with two new levels (Clan > Family) requiring sequence similarity and describing repeat motifs in collaboration with Pfam. Data growth has been addressed with improved mechanisms for browsing the classification hierarchy. A new UniProt-centric view unifies the increasingly frequent annotation of structures from identical or similar sequences. This update of RepeatsDB aligns with our commitment to develop a resource that extracts, organizes and distributes specialized information on tandem repeat protein structures., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Baculoviruses are a group of insect viruses with large circular dsDNA genomes exploited in numerous biotechnological applications, such as the biological control of agricultural pests, the expression of recombinant proteins or the gene delivery of therapeutic sequences in mammals, among others. Their genomes encode between 80 and 200 proteins, of which 38 are shared by all reported species. Thanks to multi-omic studies, there is remarkable information about the baculoviral proteome and the temporality in the virus gene expression. This allows some functional elements of the genome to be very well described, such as promoters and open reading frames. However, less information is available about the transcription termination signals and, consequently, there are still imprecisions about what are the limits of the transcriptional units present in the baculovirus genomes and how is the processing of the 3' end of viral mRNA. Regarding to this, in this review we provide an update about the characteristics of DNA signals involved in this process and we contribute to their correct prediction through an exhaustive analysis that involves bibliography information, data mining, RNA structure and a comprehensive study of the core gene 3' ends from 180 baculovirus genomes.

Organophosphorus compounds (OPs), the major pesticides used worldwide, comprise an environmental hazard due to their harmful toxicity. Aimed to develop a bioreactor to remediate OPs contaminated wastewater, bacteria isolated from contaminated soils were identified and their ability to degrade OPs assessed, resulting in two main isolates, Sphingomonas sp. and Brevundimonas sp. Their OP degrading activities were characterized in terms of temperature, pH and substrates acceptance, resulting in high degradation rates at 60 °C, pH 10 and towards bulky OPs such as coroxon, coumaphos, and chlorpyrifos. Sphingomonas sp. cells were immobilized and 75.4% degradation of 0.15 mM chlorpyrifos was achieved after 21 days by immobilized cells in batch system, while this OP was completely degraded within 17 h when the biocatalyst is settled in a packed bed bioreactor, with a reusability of 8 cycles. These results suggest the potential application of this system in the bioremediation of contaminated wastewater., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The use of microbial phosphotriesterases in the degradation of organophosphorus compounds employed as pesticides, plasticizers and petroleum additives is a sustainable alternative for bioremediation of water and soils, decontamination of particular foods and as poisoning antidote. Whole cells of six wild type microorganisms-Streptomyces phaeochromogenes, Streptomyces setonii, Nocardia corynebacterioides, Nocardia asteroides and two Arthrobacter oxydans-selected in our lab as phosphotriesterase sources, were further tested as biocatalysts in the hydrolysis of paraoxon, methyl paraoxon, methyl parathion, coroxon, coumaphos, dichlorvos and chlorpyrifos, highlighting 98% conversion of chlorpyrifos into its hydrolysis products using whole cells of S. phaeochromogenes at pH 8 and 40 °C. Immobilized whole cells and enzyme extracts were also assessed, observing as a general trend, that there is no significant variation in hydrolytic activity between them. These results suggest that according to the circumstances, immobilized whole cells (avoiding cellular disruption and centrifugation) or enzyme extracts (which can be handled more easily) could be used.

This study determined the spatial and temporal dynamics of two native neotropical species flies of forensic interest, belonging to the Lucilia (Robineau-Desvoidy) genus. The study focused on their abundance and reproductive behaviors associated with different habitats and phenological parameters. In the Province of Salta, Argentina, monthly samplings were performed over 1 yr in urban, rural, and native habitats, at morning, mid-day, and afternoon periods, controlling the oviposition of captured specimens. Environmental variables were also assessed: cloudiness, precipitation, relative humidity, temperature, and tree cover. Lucilia purpurascens (Walker) appeared to be associated with native habitats, whereas Lucilia ochricornis (Wiedemann) was mainly associated with rural habitats, exhibiting distinct habitat preferences. Two ecotones were also identified: rural-urban and rural-native, suggesting rural habitats promoted habitable conditions at its margins. Both species were recorded at the end of winter to the middle of autumn, with an initial peak in early spring, and a second peak at late summer. These peaks were associated with the highest numbers of laid eggs. Lucilia purpurascens preferred high tree coverings, whereas L. ochricornis resisted areas with intermediate sun, suggesting limited sun exposure was important. The latter was also associated with daily flight activities; during the warm season, the densest catches occurred at morning and afternoon periods, whereas during the cold season, they occurred at mid-day. Climatic variables explained 77% of variability in terms of abundance and oviposition. Synergistic effects were observed between these variables, suggesting that these variables conditioned insect distribution and reproduction, and not just temperature per se., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Cladribine (2-chloro-2'-deoxy-β-d-adenosine) is a 2'-deoxyadenosine analogue, approved by the FDA for the treatment of hairy cell leukemia and more recently has been proved for therapeutic against many autoimmune diseases as multiple sclerosis. The biosynthesis of this compound using Thermomonospora alba CECT 3324 as biocatalyst is herein reported. This thermophilic microorganism was successfully entrapped in polyacrylamide gel supplemented with nanoclays such as bentonite. The immobilized biocatalyst (T. alba-Ac-Bent 1.00 %), was able to biosynthesize cladribine with a conversion of 89 % in 1 h of reaction and retains its activity for more than 270 reuses without significantly activity loss, showing better operational stability and mechanical properties than the natural matrix. A microscale assay using the developed system, could allow the production of at least 181 mg of cladribine in successive bioprocesses., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this work was to assess the temporal patterns in the community composition of phytoplankton (PCC) and bacterioplankton (BCC) in two interconnected and hypertrophic Pampean shallow lakes in Argentina. Factors shaping their community dynamics and community temporal covariations were also analysed. We performed 4 years of seasonal samplings (2012-2016) and communities were studied by the Utermöhl approach (PCC) and Illumina MiSeq sequencing (BCC). We found marked seasonal variations in both communities and inter-annual variations with decreasing microbial community similarities during the study. We also observed covariation in community-level dynamics among PCC and BCC within and between shallow lakes. The within-lake covariations remained positive and significant, while controlling for the effects of intrinsic (environmental) and extrinsic (temporal and meteorological) factors, suggesting a community coupling mediated by intrinsic biotic interactions. Algal-bacterial associations between different taxa of phytoplankton and bacterioplankton within each lake were also found. PCC was mainly explained by pure regional extrinsic (17-21%) and intrinsic environmental (8-9%) factors, while BCC was explained by environmental (8-10%) and biotic interactions with phytoplankton (7-8%). Our results reveal that the influence of extrinsic regional factors can be channeled to bacterioplankton through both environmental (i.e. water temperature) and phytoplankton effects., (© The Author(s) 2020. Published by Oxford University Press on behalf of FEMS.)

A novel IDA-LaNDT derivative was able to reach the highest productivity in the biosynthesis of a well-known antitumoral agent called decitabine. However, the combination of two simple and inexpensive techniques such as ionic absorption and gel entrapment with the incorporation of a bionanocomposite such as bentonite significantly improved the stability of this biocatalyst. These modifications allowed the enhancement of storage stability (for at least 18 months), reusability (400 h of successive batches without significant loss of its initial activity), and thermal and solvent stability with respect to the non-entrapped derivative. Moreover, reaction conditions were optimized by increasing the solubility of 5-aza by dilution with dimethylsulfoxide. Therefore, a scale-up of the bioprocess was assayed using the developed biocatalyst, obtaining 221 mg/L·h of DAC. Finally, green parameters were calculated using the nanostabilized biocatalyst, whose results indicated that it was able to biosynthesize DAC by a smooth, cheap, and environmentally friendly methodology.

The propensity account of fitness intends to solve the classical tautologicity issue by identifying fitness with a disposition, the ability to survive and reproduce. As proponents recognized early on, this account requires operational independence from actual reproductive success to avoid circularity and vacuousness charges. They suggested that operational independence is achieved by measuring fitness values through optimality models. Our goal in this article is to develop this suggestion. We show that one plausible procedure by which these independent operationalizations could be thought to take place, and which is in accordance with what is said in the optimality literature, is unsound. We provide two independent lines of reasoning to show this. We then provide a sketch of a more adequate account of the role of optimality models in evolutionary contexts and draw some consequences.

Pseudomonas donghuensis strain SVBP6, an isolate from an agricultural plot in Argentina, displays a broad-spectrum and diffusible antifungal activity, which requires a functional gacS gene but could not be ascribed yet to known secondary metabolites typical of Pseudomonas biocontrol species. Here, we report that Tn5 mutagenesis allowed the identification of a gene cluster involved in both the fungal antagonism and the production of a soluble tropolonoid compound. The ethyl acetate extract from culture supernatant showed a dose-dependent inhibitory effect against the phytopathogenic fungus Macrophomina phaseolina. The main compound present in the organic extract was identified by spectroscopic and X-ray analyses as 7-hydroxytropolone (7HT). Its structure and tautomerism was confirmed by preparing the two key derivatives 2,3-dimethoxy- and 2,7-dimethoxy-tropone. 7HT, but not 2,3- or 2,7-dimethoxy-tropone, mimicked the fungal inhibitory activity of the ethyl acetate extract from culture supernatant. The activity of 7HT, as well as its production, was barely affected by the presence of up to 50 μM added iron (Fe +2 ). To summarize, P. donghuensis SVBP6 produces 7HT under the positive control of the Gac-Rsm cascade and is the main active metabolite responsible for the broad-spectrum inhibition of different phytopathogenic fungi., (© 2020 Society for Applied Microbiology and John Wiley & Sons Ltd.)

A stable biocatalyst with magnetic properties based on immobilized Lactobacillus animalis ATCC 35,046 to obtain 2-chloroadenine-2'-deoxyriboside, known as cladribine, is reported for the first time. This nucleoside analogue is an antitumor agent used in the treatment of a wide variety of types of leukemia. In this study, an eco-compatible and alternative bioprocess to obtain cladribine was developed. Product conversion was close to 90% at 2 h in optimized nonconventional reaction media. The microscale biosynthesis of the compound of interest afforded a total productivity close to 370 mg/L/h in the presence of DMSO, and it was stable at least for 30 days in storage conditions.

Group A rotaviruses (RVA) are the most frequent etiological agents causing severe diarrhea in infants and surveillance of genotype, and genetic characteristics of circulating strains are necessary in order to evaluate vaccine programs. The objectives of this work were to describe G and P genotype from 2012 through 2014 in Buenos Aires, Argentina completing an overview of 19 years of genotype surveillance in our region and to characterize an emerging G1P[8] strain associated with severe cases and five fatalities in 2014. We performed genotyping by RT-PCR. The sequencing of several genes, phylogenetic analyses, and comparative epidemiological data were used to know the origin and phylogenetic relationships of the emerging G1P[8] strain. Along with this report, 19 years of continuous RVA genotype surveillance in Argentina in the pre-vaccine era was covered. During the last year of this surveillance, 2014, a significantly increased incidence of RVA associated gastroenteritis was related to the reemergence of G1P[8] strains, being these ones detected in low frequency in the last nine years. Interestingly, the patients affected were significantly older when compared with those from the last six seasons. Additionally, phylogenetic analysis of several genes infer that these G1P[8] strains were closely related to Asian strains circulating during 2012 and 2013. In addition to this, the suggested extra continental origin for the 2014 G1P[8] strains and the very low circulation of G1 type during nine years probably explain the increased incidence and severity in the gastroenteritis cases and the particular epidemiologic characteristics. In conclusion, this work gives us a whole panorama of the pre-vaccine era of the RVA molecular epidemiology in the most populated region of Argentina. In this way, this work inspires us to continue with this type of studies in the post-vaccination era., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Thiol peroxidase from Escherichia coli (EcTPx) is a peroxiredoxin that catalyzes the reduction of different hydroperoxides. During the catalytic cycle of EcTPx, the peroxidatic cysteine (C P ) is oxidized to a sulfenic acid by peroxide, then the resolving cysteine (C R ) condenses with the sulfenic acid of C P to form a disulfide bond, which is finally reduced by thioredoxin. Purified EcTPx as dithiol and disulfide behaves as a monomer under near physiological conditions. Although secondary structure rearrangements are present when comparing different redox states of the enzyme, no significant differences in unfolding free energies are observed under reducing and oxidizing conditions. A conformational change denominated fully folded (FF) to locally unfolded (LU) transition, involving a partial unfolding of αH2 and αH3, must occur to enable the formation of the disulfide bond since the catalytic cysteines are 12 Å apart in the FF conformation of EcTPx. To explore this process, the FF → LU and LU → FF transitions were studied using conventional molecular dynamics simulations and an enhanced conformational sampling technique for different oxidation and protonation states of the active site cysteine residues C P and C R . Our results suggest that the FF → LU transition has a higher associated energy barrier than the refolding LU → FF process in agreement with the relatively low experimental turnover number of EcTPx. Furthermore, in silico designed single-point mutants of αH3 enhanced locally unfolding events, suggesting that the native FF interactions in the active site are not evolutionarily optimized to fully speed-up the conformational transition of wild-type EcTPx.

Water-solid interactions were explored in purified and freeze-dried Prosopis alba exudate gum as approach to get a deeper insight of structural and functional aspects of this novel biomaterial. Particularly, the study of water-binding properties combined with glass transition temperatures allowed obtaining interesting theoretical data for practical applications. The Guggenheim-Anderson-de Boer (GAB) and Generalized D'Arcy and Watt (GDW) models were applied to describe the sorption behavior and thermodynamic properties of the studied gum. The study of the relationship between relative humidity, water content and thermal transitions allowed to characterize the material in terms of water plasticizing susceptibility as well as define the suitable storage conditions that guarantee the quality, safety and physical stability of P. alba gum. Obtained results contribute to the characterization of a non-conventional exudate gum with great potential for its use in different food industry applications., Competing Interests: Declaration of Competing Interest The authors declared that there is no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

The biocatalyzed synthesis of purine nucleosides and their analogs is a case widely studied due to the high pharmaceutical interest of these compounds, providing the whole-cell biocatalysts, a useful tool for this purpose. Vidarabine and fludarabine are commercial examples of expensive bioactive nucleosides that can be prepared using a microbial transglycosylation approach. Citrobacter koseri whole-cells immobilized on agarose beads proved to be an interesting option to transform this biotransformation in a preparative process. The entrapment matrix provided a useful and resistant multipurpose biocatalyst regarding its stability, mechanical strength, microbial viability and reuse. Immobilized biocatalyst retained the initial activity for up to 1 year storage and after 10 years, the biocatalyst did not show cell leaking and still exhibited residual activity. In addition, the biocatalyst could be reused in batch 68 times keeping up to 50% of the initial biocatalytic activity and for at least 124 h in a continuous process.

Alpha-amylase is frequently used in technologies that require its immobilization, stabilization or encapsulation. Polyacrylic acid is a very suitable polymer for these purposes because it can bind to enzymes and then be released under certain conditions without altering the functional capacity of enzymes. The consequences produced by polyacrylic acid on alpha-amylase structure and function have been investigated through various techniques. Calorimetric measurements allowed examining the nature of the binding reaction, stoichiometry and affinity, while spectroscopic techniques provided additional information about functional and structural perturbations of the enzyme. Isothermal titration calorimetry (ITC) revealed a mixed interaction and a binding model with a large number of molecules of protein per molecule of polyacrylic acid. One the one hand circular dichroism (CD) spectroscopy showed that alpha-amylase loses its secondary structure in the presence of increasing concentrations of polyacrylic acid, while it is stabilized by the polyelectrolyte at low pH. On the other hand, fluorescence spectra revealed that the three-dimensional enzyme structure was not affected in the microenvironment of tryptophan residues. Differential scanning calorimetry (DSC) thermograms showed that only one domain of alpha-amylase is affected in its conformational stability by the polymer. The unfolding process proved to be partially reversible. Finally, the enzyme retained more than 90 % of its catalytic activity even in excess of the polymer., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

We report the synthesis and biological evaluation of a ternary copper complex, [Cu(5HTP)(phen)(H 2 O)](NO 3 ).2H 2 O, with the antioxidant agent 5-hydroxytryptophan (5-HTP) and phenanthroline (phen, added to improve its lipophilicity and membrane transport). The crystal structure of the complex was determined by X-ray diffraction methods. The complex showed antioxidant, antimicrobial, antitumor and antimetastatic properties with an adequate safety profile. The interaction of the metal with phen promotes cellular copper accumulation and cytotoxicity on human lung A549 cell line (IC 50  = 3.6 μM). Furthermore, the viability of the normal human fetal lung fibroblast cell line (MRC-5) is not altered by the complex. An oxidative stress mechanism for the anticancer effect has been determined: cellular increase of reactive oxygen species (ROS), decrease of the glutathione (GSH) and oxidized GSH (GSSG) ratio and alteration of the mitochondrial potential. The complex also displays antimetastatic activities with inhibition of cell adhesion, invasion and migration. It has not mutagenic behavior and no toxicity on Artemia salina indicating its potential to act as an effective and safety antimicrobial and antitumor drug., Competing Interests: Declaration of competing interest None., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This article focuses on the impact of chemical composition and structural properties of tofu-whey concentrates on their surface behavior at the air/water interface and foaming properties in acid medium. Liquid tofu-whey (pH 5.6 ± 0.1) was concentrated at three combinations of temperature and pressure (50 °C-8.0 kPa, 65 °C-16.0 kPa and 80 °C-24.0 kPa), with further dialysis (4 °C, 48 h) and freeze-drying, giving the samples TWC50, TWC65 and TWC80, respectively. The increase of temperature during the concentration step promoted the enrichment of the concentrates in crude protein and calcium, without appreciable changes in the yield, the carbohydrate content and the polypeptide composition. For TWC80, the increase the degree of glycosylation and the intensity of the hydrophobic effect promoted the decrease of molecular flexibility and the formation of compact aggregates mediated by disulfide bridges as was evidenced by tricine-SDS-PAGE, TGA and FTIR assays. These structural differences have a pronounced impact of the pH-dependence of turbidity and solubility of protein and polysaccharides. At pH 4.0 all concentrates evidenced a ζ-potential close to zero, which enhanced their foam ability (overrun >1500%). Nevertheless, at this pH, TWC80 showed both the highest carbohydrate-to-protein mass ratio in the soluble fraction (>1.8) and foam stability (FS). Thus, the improvement of FS at pH 4.0 would be associated to the effective adsorption of compact rich-in-protein aggregates at the air/water interface and the higher content of soluble polysaccharides in the bulk phase. These findings are relevant for the application of tofu-whey concentrates in acidic dispersed systems, such as foams and aerated food emulsions., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

In 1996, a diagnostic study performed in a 16-ha field located in Buenos Aires Province (Argentina), where a chemical industry produced 1,2,3,4,5,6-hexachlorocyclohexane (HCH) from 1960 to 1978, showed contamination with HCH ranging from 10 to 20,000 mg kg -1  dry soil (706.4 mg kg -1 average). For remediation purposes, a forestation plan was put into practice in 1997 employing approximately 12,300 Eucalyptus dunnii seedlings which by 2016 where fully grown into trees that formed a forest where local fauna can be found. Midterm analysis done in 2005, when E. dunnii trees had developed into 8-10 m high trees, indicated that HCH was incorporated into leaves and logs and soil phytoremediation was progressing. Final quantitation analysis of HCH in soil performed in 2016 demonstrated that the 97.2% of the field area was effectively decontaminated with 98.1% overall average efficiency. Thus, this work is the first global example of a successful employment of E. dunnii trees for HCH phytoremediation purposes at field scale. These results may encourage other researchers to test the ability of E. dunnii to phytoremediate soils contaminated with other chlorinated compounds like polychlorinated biphenyls (PCBs), polychlorinated dibenzodioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs).

Aims: G protein-coupled receptor (GPCR) kinases (GRKs) are mainly involved in the desensitization of GPCRs. Among them, GRK2 has been described to be upregulated in many pathological conditions and its crucial role in cardiac hypertrophy, hypertension, and heart failure promoted the search for pharmacological inhibitors of its activity. There have been several reports of potent and selective inhibitors of GRK2, most of them directed to the kinase domain of the protein. However, the homologous to the regulator of G protein signaling (RH) domain of GRK2 has also been shown to regulate GPCRs signaling. Herein, we searched for potential inhibitors of receptor desensitization mediated by RH domain of GRK2., Materials and Methods: We performed a docking-based virtual screening utilizing the crystal structure of GRK2 to search for potential inhibitors of the interaction between GRK2 and Gαq protein. To evaluate the biological activity of compounds we measured, calcium response of histamine H1 receptor (H1R) using Fura-2AM dye and H1R internalization by saturation binding experiments in A549 cells. GRK2(45-178)GFP translocation was determined in HeLa cells through confocal fluorescence imaging., Key Findings: We identified inhibitors of GRK2 able to reduce the RH mediated desensitization of the histamine H1 receptor and GRK2 translocation to plasma membrane. Also candidates presented adequate lipophilia and cytotoxicity profile., Significance: We obtained compounds with the ability of reducing RH mediated actions of GRK2 that can be useful as a starting point in the development of novel drug candidates aimed to treat pathologies were GRK2 plays a key role., (Copyright © 2019 Elsevier Inc. All rights reserved.)

This article elaborates on McShea and Brandon's idea that drift is unlike the rest of the evolutionary factors because it is constitutive rather than imposed on the evolutionary process. I show that the way they spelled out this idea renders it inadequate and is the reason why it received some (good) objections. I propose a different way in which their point could be understood, that rests on two general distinctions. The first is a distinction between the underlying mathematical apparatus used to formulate a theory and a concept proposed by that theory. With the aid of a formal reconstruction of a population genetic model, I show that drift belongs to the first category. That is, that drift is constitutive of population genetics in the same sense that multiplication is constitutive in classical mechanics, or that circle is constitutive in Ptolemaic astronomy. The second distinction is between eliminating a concept from a theory and setting its value to zero. I will show that even though drift can be set to zero just like the rest of the evolutionary factors (as others have noted in their criticism of McShea and Brandon), eliminating drift is much harder than eliminating those other factors, since it would require changing the entire mathematical apparatus of standard population genetic theory. I conclude by drawing some other implications from the proposed formal reconstruction.

Dengue fever is a mosquito-borne viral disease that has become a major public health concern worldwide. This disease presents with a wide range of clinical manifestations, from a mild cold-like illness to the more serious hemorrhagic dengue fever and dengue shock syndrome. Currently, neither an approved drug nor an effective vaccine for the treatment are available to fight the disease. The envelope protein (E) is a major component of the virion surface. This protein plays a key role during the viral entry process, constituting an attractive target for the development of antiviral drugs. The crystal structure of the E protein reveals the existence of a hydrophobic pocket occupied by the detergent n-octyl-β-d-glucoside (β-OG). This pocket lies at the hinge region between domains I and II and is important for the low pH-triggered conformational rearrangement required for the fusion of the virion with the host's cell. Aiming at the design of novel molecules which bind to E and act as virus entry inhibitors, we undertook a de novo design approach by "growing" molecules inside the hydrophobic site (β-OG). From more than 240000 small-molecules generated, the 2,4 pyrimidine scaffold was selected as the best candidate, from which one synthesized compound displayed micromolar activity. Molecular dynamics-based optimization was performed on this hit, and thirty derivatives were designed in silico, synthesized and evaluated on their capacity to inhibit dengue virus entry into the host cell. Four compounds were found to be potent antiviral compounds in the low-micromolar range. The assessment of drug-like physicochemical and in vitro pharmacokinetic properties revealed that compounds 3e and 3h presented acceptable solubility values and were stable in mouse plasma, simulated gastric fluid, simulated intestinal fluid, and phosphate buffered saline solution., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

The characteristics and quality of home-made dry cured sausages can be recognized and associated with the region of origin. The characteristics of this type of sausages result from the superficial mycobiota that spontaneously colonizes the products. The aim of this study was to identify the house mycobiota associated with home-made dry cured sausages from different localities of Argentina and characterize the populations of Penicillium nalgiovense present by morphological and biochemical markers. To this end, 79 samples were collected from 10 localities of three main producing regions (Buenos Aires, Córdoba and La Pampa provinces). A total of 196 isolates belonging to six genera and 17 species were obtained. The predominant genus was Penicillium (134 of the isolates) and the predominant species was P. nalgiovense (108 isolates). The isoenzyme patterns of α-esterase (α-EST; EC 3.1.1.1) and Malate dehydrogenase NADP + (MDHP; EC 1.1.1.40) were characterized in 48 of these isolates (ten from Colonia Caroya, ten from Oncativo, ten from Tandil, nine from Mercedes and nine from La Pampa). A total of 26 bands were observed: 17 for α-EST and 9 for MDHP. α-EST was the most polymorphic isoenzyme, whereas MDPH presented no polymorphism. The results were subjected to numerical analysis. Cluster analysis revealed the formation of two groups: Group I formed by 24 isolates from Córdoba and Buenos Aires provinces and Group II with 24 isolates from La Pampa and Buenos Aires province. These data suggest the existence of morphological and biochemical variations among P. nalgiovense populations with different geographical origin., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The ectomycorrhizal symbiosis is a predominant tree-microbe interaction in forest ecosystems sustaining tree growth and health. Its establishment and functioning implies a long-term and intimate relationship between the soil-borne fungi and the roots of trees. Mycorrhiza-induced Small-Secreted Proteins (MiSSPs) are hypothesized as keystone symbiotic proteins, required to set up the symbiosis by modifying the host metabolism and/or building the symbiotic interfaces. L. bicolor MiSSP8 is the third most highly induced MiSSPs in symbiotic tissues and it is also expressed in fruiting bodies. The MiSSP8-RNAi knockdown mutants are strongly impaired in their mycorrhization ability with Populus, with the lack of fungal mantle and Hartig net development due to the lack of hyphal aggregation. MiSSP8 C-terminus displays a repetitive motif containing a kexin cleavage site, recognized by KEX2 in vitro. This suggests MiSSP8 protein might be cleaved into small peptides. Moreover, the MiSSP8 repetitive motif is found in other proteins predicted secreted by both saprotrophic and ectomycorrhizal fungi. Thus, our data indicate that MiSSP8 is a small-secreted protein involved at early stages of ectomycorrhizal symbiosis, likely by regulating hyphal aggregation and pseudoparenchyma formation., (© 2019 Society for Applied Microbiology and John Wiley & Sons Ltd.)

Neglected tropical diseases (NTDs) constitute a group of infectious diseases prevalent in countries with tropical and subtropical climate that affect the poorest individuals and produce high chronic disability associated with serious problems for the health system and socioeconomic development. Chagas disease or American trypanosomiasis is included on the NTDs list. However, even though this disease affects more than 10 million people, mostly in Latin America, causing the death of over 10,000 people every year, only two drugs are approved for its treatment, benznidazole and nifurtimox. These antiparasitic agents were developed almost half a century ago and present several biopharmaceutical disadvantages such as low aqueous solubility and permeability limiting their bioavailability. In addition, both therapeutic agents are available only as tablets and a liquid pediatric formulation is still lacking. Therefore, novel pharmaceutical strategies to optimize the pharmacotherapy of Chagas disease are urgently required. In this regard, nanotechnological approaches may be a crucial alternative for the delivery of both drugs ensuring an effective pharmacotherapy although the successful bench-to-bedside translation remains a major challenge. The present work reviews in detail the formulation and in-vitro/in-vivo analysis of different nanoformulations of nifurtimox and benznidazole in order to enhance their solubility, dissolution, bioavailability and trypanocidal activity., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The objective of this work was to deepen on the study of functional properties of the phytochemicals present in Prosopis alba exudate gum (G), as well as to rule out possible adverse effects of some of its components. Commonly employed purification methods were compared. Filtration prevents further loss of potentially bioactive compounds. The filtrated gum showed a higher concentration of phenolics, flavonoids and tannins than arabic gum, which was correlated with better in vitro antioxidant properties. Particularly, tannins, commonly considered as toxic compounds in exudate gums, were found in lower concentration than in others gums obtained from genus Prosopis and Acacia. The toxicological evaluation performed on rats did not show symptoms of intoxication associated with the administration of the gum. These results provide useful evidence to support the potential use of G as a safe functional food additive with the added benefit of taking advantage of a non-exploited natural resource., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The impact of lead (Pb) pollution on native communities of arbuscular mycorrhizal fungi (AMF) was assessed in soil samples from the surroundings of an abandoned Pb smelting factory. To consider the influence of host identity, bulk soil surrounding plant roots soil samples of predominant plant species (Sorghum halepense, Bidens pilosa, and Tagetes minuta) growing in Pb-polluted soils and in an uncontaminated site were selected. Molecular diversity was assessed by sequencing the 18S rDNA region with primers specific to AMF (AMV4.5NF/AMDGR) using Illumina MiSeq. A total of 115 virtual taxa (VT) of AMF were identified in this survey. Plant species did not affect AMF diversity patterns. However, soil Pb content was negatively correlated with VT richness per sample. Paraglomeraceae and Glomeraceae were the predominant families while Acaulosporaceae, Ambisporaceae, Archaeosporaceae, Claroideoglomeraceae, Diversisporaceae, and Gigasporaceae were less abundant. Acaulosporaceae and Glomeraceae were negatively affected by soil Pb, but Paraglomeraceae relative abundance increased under increasing soil Pb content. Overall, 26 indicator taxa were identified; four of them were previously reported in Pb-polluted soils (VT060; VT222; VT004; VT380); and five corresponded to cultured spores of Scutellospora castaneae (VT041), Diversispora spp. and Tricispora nevadensis (VT060), Diversispora epigaea (VT061), Glomus proliferum (VT099), and Gl. indicum (VT222). Even though AMF were present in Pb-polluted soils, community structure was strongly altered via the differential responses of taxonomic groups of AMF to Pb pollution. These taxon-specific differences in tolerance to soil Pb content should be considered for future phytoremediation strategies based on the selection and utilization of native Glomeromycota.

Chronic exposure to inorganic arsenic (As) is associated with numerous adverse effects. Argentina is one of the countries affected by arsenicism; however, there are few studies that evaluate inorganic As exposure and its effects on child population. The aim of this study is to evaluate exposure to As through water and food in child populations living in the provinces of Santiago del Estero and Chaco (n = 101), and to determine the impact of this exposure analysing biomarkers of exposure (urine and hair As contents) and effect [8-oxo-7,8-dihydro-2'-deoxyguanosine (8-OHdG)]. The populations selected live in three areas with different levels of As in the drinking water (Santa Teresa de Carballo, 0.925 mg/L; Taco Pozo, 0.210 mg/L; Jumi Pozo, 0.016 mg/L). The As intakes through water and food are especially high in the areas with the greatest As exposure (Santa Teresa de Carballo, 1575 ± 8 μg/day; Taco Pozo, 386 ± 8 μg/day; Jumi Pozo, 39 ± 1 μg/day). The total As contents in most of the samples of hair (0.11-13.11 mg/kg) and urine (31-4258 μg/g creatinine) are higher than the reference values (hair: 1 mg/kg; urine: 50 μg/g creatinine). The increase in the level of As exposure alters the profile of metabolites in urine, with a decrease of dimethylarsinic acid (10%) and an increase in the percentages of monomethylarsonic acid (4%) and inorganic As (6%). The results also show high values of 8-OHdG (3.7-37.8 μg/g creatinine), a oxidative DNA damage marker, in the two areas with greater As exposure., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Benznidazole (BNZ) is the drug of choice for the treatment of Chagas disease in many countries. However, its low water solubility produces low and/or variable oral bioavailability. Thus, the aim of this work was to formulate micro- and nanoparticles based on Eudragit ® RS PO and Eudragit ® RL PO as a convenient approach to increase the dissolution rate of BNZ. The microparticles were obtained by means of spray-drying process while the nanoparticles were prepared through the nanoprecipitation technique and further freeze-drying. The results indicated that nanoparticles were obtained in 86% yield while microparticles were obtained in 68% yield. In both cases, the encapsulation efficiency of particles was greater than 78% while drug loading capacity was nearly 24% w/w and 18% w/w, after spray-drying and freeze-drying procedures, respectively. Images of scanning electron microscopy showed that the particles obtained by spray-drying and freeze-drying were in the micrometer and nanometer scale, respectively. FT-IR spectra of BNZ-loaded particles obtained by both methods showed characteristic bands of BNZ confirming that part of drug remained on their surface. Thermal analysis revealed that the drug crystallinity after both methods decreased. Physical stability evaluation of the nanoparticles confirmed that Pluronic ® F68 was suitable to keep the particles size in a range of 300 nm after 70 days storage at 4 ± 2 °C. In-vitro release studies showed increased dissolution rate of drug from the particles obtained by both methods respect to untreated BNZ. The kinetics of drug release in acid media followed the Higuchi kinetics indicating drug diffusion mechanism from particles., (Copyright © 2019 Elsevier B.V. All rights reserved.)

[Formula: see text]-Lactamases (penicillinases) facilitate bacterial resistance to antibiotics and are excellent theoretical and experimental models in protein structure, dynamics and evolution. Bacillus licheniformis exo-small penicillinase (ESP) is a Class A [Formula: see text]-lactamase with three tryptophan residues located one in each of its two domains and one in the interface between domains. The conformational landscape of three well-characterized ESP Trp[Formula: see text]Phe mutants was characterized in equilibrium unfolding experiments by measuring tryptophan fluorescence, far-UV CD, activity, hydrodynamic radius, and limited proteolysis. The Trp[Formula: see text]Phe substitutions had little impact on the native conformation, but changed the properties of the partially folded states populated at equilibrium. The results were interpreted in the framework of modern theories of protein folding.

The conformations accessible to proteins are determined by the inter-residue interactions between amino acid residues. During evolution, structural constraints that are required for protein function providing biologically relevant information can exist. Here, we studied the proportion of sites evolving under structural constraints in two very different types of ensembles, those coming from ordered and disordered proteins. Using a structurally constrained model of protein evolution, we found that both types of ensembles show comparable, near 40%, number of positions evolving under structural constraints. Among these sites, ~68% are in disordered regions and ~57% of them show long-range inter-residue contacts. Also, we found that disordered ensembles are redundant in reference to their structurally constrained evolutionary information and could be described on average with ~11 conformers. Despite the different complexity of the studied ensembles and proteins, the similar constraints reveal a comparable level of selective pressure to maintain their biological functions. These results highlight the importance of the evolutionary information to recover meaningful biological information to further characterize conformational ensembles., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

5-Hydroxy-L-tryptophan (5-HTP) is a serotonin pathway metabolite of L-tryptophan in the brain. In the knowledge that the biological properties of some compounds can be modified upon metal complexation, a new solid metal complex, [Cu(5-hydroxytryptophan) 2 ].H 2 O (Cu5HTP), has been synthesized and characterized to analyze the modification of some biological properties. The conformational investigations (optimized in gas phase at B3LYP/6-311G** theory level) suggest the coexistence of two conformers of Cu5HTP with cis- and trans- arrangements of the amino acids in the equatorial plane. The trans- Cu5HTP1 complex is the most stable conformer. The complexation led to an enhancement of the antioxidant properties of the ligand. The metal complex also improved the anticancer behavior of the ligand (tested in cancer cell lines derived from human lung (A549), cervix (HeLa) and colon (HCT-116)). It did not show toxicity against either the non-malignant human lung fibroblast (MRC-5) cell line or Artemia salina and did not behave as mutagenic agent (Ames test). Cellular reactive oxygen species production may be one of the possible mechanisms of action. Besides, the metal complex exerted neuroprotective action on cortical neurons from embryonic 18 days rats exposed to glutamate., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Saint Louis encephalitis virus (SLEV) and West Nile virus (WNV) are two of the major causes of arboviral encephalitis in the Americas. The co-circulation of related flaviviruses in the Americas and prior vaccination against flaviviruses pose problems to the diagnostic specificity of serological assays due to the development of cross-reactive antibodies. An accurate diagnosis method capable of differentiating these related viruses is needed. NS1 is a glycosylated, nonstructural protein, of about 46 kDa which has a highly conserved structure. Anti-NS1 antibodies can be detected within 4-8 days after the initial exposure and NS1 is the least cross-reactive of the flaviviral antigens. This study was aimed to generate SLEV and WNV NS1 recombinants proteins for the development of a flavivirus diagnostic test. Local Argentinian isolates were used as the source of NS1 gene cloning, expression, and purification. The protein was expressed in Escherichia coli as inclusion bodies and further purified by metal-chelating affinity chromatography (IMAC) under denaturing conditions. Human sera from SLEV and WNV positive cases showed reactivity to the recombinant NS1 proteins by western blot. The unfolded NS1 proteins were also used as immunogens. The polyclonal antibodies elicited in immunized mice recognized the two recombinant proteins with differential reactivity., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Mammalian frataxin is a small mitochondrial protein involved in iron sulfur cluster assembly. Frataxin deficiency causes the neurodegenerative disease Friedreich's Ataxia. Valuable knowledge has been gained on the structural dynamics of frataxin, metal-ion-protein interactions, as well as on the effect of mutations on protein conformation, stability and internal motions. Additionally, laborious studies concerning the enzymatic reactions involved have allowed for understanding the capability of frataxin to modulate Fe-S cluster assembly function. Remarkably, frataxin biological function depends on its interaction with some proteins to form a supercomplex, among them NFS1 desulfurase and ISCU, the scaffolding protein. By combining multiple experimental tools including high resolution techniques like NMR and X-ray, but also SAXS, crosslinking and mass-spectrometry, it was possible to build a reliable model of the structure of the desulfurase supercomplex NFS1/ACP-ISD11/ISCU/frataxin. In this chapter, we explore these issues showing how the scientific view concerning frataxin structure-function relationships has evolved over the last years.

Promiscuous behaviour in proteins and enzymes remains a challenging feature to understand the structure-function relationship. Here we present ProtMiscuity, a manually curated online database of proteins showing catalytic promiscuity. ProtMiscuity contains information about canonical and promiscuous activities comprising 88 different reactions in 57 proteins from 40 different organisms. It can be searched or browsed by protein names, organisms and descriptions of canonical and promiscuous reactions. Entries provide information on reaction substrates, products and kinetic parameters, mapping of active sites to sequence and structure and links to external resources with biological and functional annotations. ProtMiscuity could assist in studying the underlying mechanisms of promiscuous reactions by offering a unique and curated collection of experimentally derived data that is otherwise hard to find, retrieve and validate from literature., (© The Author(s) 2019. Published by Oxford University Press.)

In the present study, we evaluated the transcriptional response of four stress-related genes in three Oenococcus oeni strains after acclimation at two different temperatures. Gene expression was analyzed at time zero and after 48 h acclimation at 18 and 21 °C. After the acclimation period cells were inoculated into sterile Pinot noir wine and MLF was followed for 25 days to investigate if different acclimation temperatures could influence cell survival and MLF performance. L-malic acid consumption, population survival, and transcriptional behavior were different upon the acclimation temperature. rmlB and hsp20 genes presented a considerable increase in their expression level when strains were acclimated at 18 °C particularly in the psychrotrophic strains UNQOe19 and UNQOe4 isolated from Patagonian Pinot noir wine in comparison with the control strain (ATCC 27310). The increase in rmlB and hsp20 expression could account for the better survival of these strains in Pinot noir in comparison with the control strain. In addition, Patagonian populations acclimated at 18 °C were able to consume a higher percentage of L-malic acid in comparison with cells acclimated at 21 °C. Our results suggest that gene expression analysis of cells acclimated at sub-optimal temperatures could benefit the selection of psychrotrophic strains aimed as starter cultures.

Bacteria of the Azospirillum and Pseudomonas genera are ubiquitous members of the rhizosphere, where they stimulate plant growth. Given the outstanding capacity of pseudomonads to antagonize other microorganisms, we analyzed the interaction between these two bacterial groups to identify determinants of their compatibility. We could establish that, when in direct contact, certain Pseudomonas strains produce lethality on Azospirillum brasilense cells using an antibacterial type 6 secretion system. When analyzing the effect of Pseudomonas spp. diffusible metabolites on A. brasilense growth on King's B medium, we detected strong inhibitory effects, mostly mediated by siderophores. On Congo Red medium, both inhibitory and stimulatory effects were induced by unidentified compounds. Under this condition, Pseudomonas protegens CHA0 produced a Gac/Rsm-regulated antibiotic which specifically inhibited A. brasilense Sp7 but not Sp245. This effect was not associated with the production of 2,4-diacetylphloroglucinol. The three identified antagonism determinants were also active in vivo, producing a reduction of viable cells of A. brasilense in the roots of wheat seedlings when co-inoculated with pseudomonads. These results are relevant to the understanding of social dynamics in the rhizosphere and might aid in the selection of strains for mixed inoculants.

Background: Thiazolidinone derivatives show inhibitory activity (IC 50 ) against the Toxoplasma gondii parasite, as well as high selectivity with high therapeutic index. To disclose the target proteins of the thiazolidinone core in this parasite, we explored in silico the active sites of different T. gondii proteins and estimated the binding-free energy of reported thiazolidinone molecules with inhibitory effect on invasion and replication of the parasite inside host cells. This enabled us to describe some of the most suitable structural characteristics to design a compound derived from the thiazolidinone core., Results: The best binding affinity was observed in the active site of kinase proteins, we selected the active site of the T. gondii ROP18 kinase, because it is an important factor for the virulence and survival of the parasite. We present the possible effect of a derivative of thiazolidinone core in the active site of T. gondii ROP18 and described some characteristics of substituent groups that could improve the affinity and specificity of compounds derived from the thiazolidinone core against T. gondii., Conclusions: The results of our study suggest that compounds derived from the thiazolidinone core have a preference for protein kinases of T. gondii, being promising compounds for the development of new drugs with potential anti-toxoplasmosis activity. Our findings highlight the importance of use computational studies for the understanding of the action mechanism of compounds with biological activity.