Antimicrobial action of synthetic peptides towards wine spoilage yeasts

The antimicrobial action of selected short synthetic peptides against wine spoilage yeasts such as Cryptococcus albidus, Dekkera bruxellensis, Pichia membranifaciens, Saccharomyces cerevisiae, Zygosaccharomyces bailii and Zygosaccharomyces bisporus has been examined. Peptides analyzed include nine sequence-related antifungal hexapeptides (PAFs) previously developed by a combinatorial approach, and two representative lactoferricin B (LfcinB)-derived peptides. Different peptides had distinct activity profiles. In vitro assays identified the peptides PAF26, PAF36, and LfcinB(17-31), as having growth inhibitory properties towards several of the yeasts at low micromolar concentrations. Z. bailii and Z. bisporus were the most sensitive yeasts. In addition to their fungistatic activity, the three peptides showed fungicidal properties towards Z. bailii, Z. bisporus, and S. cerevisiae in laboratory growth medium. Remarkably, only LfcinB(17-31) against Z. bisporus had inhibitory and fungicidal properties in wine at the concentrations assayed, showing that the antimicrobial action of each peptide is dependent on both the food matrix and the target micro-organism. Lack of fungicidal activity of peptides against Z. bailii in wine is related to the presence of salt ions other than divalent cations. On the contrary, fungicidal activity of LfcinB(17-31) towards Z. bisporus was not significantly affected by wine salts. Our data identify a bioactive peptide from natural origin with potential use against the food spoilage yeast Z. bisporus, and indicate that the application of antimicrobial peptides in wine preservation deserves further investigation.     

Concentration changes of organochlorine compounds and polybromodiphenyl ethers during metamorphosis of aquatic insects

The role of insect larvae and pupae as sources of organochlorine compounds (OCs) and polybromodiphenyl ethers (PBDEs) in freshwater food webs for high predators such as fish is evaluated. Trichoptera and diptera have been taken as organisms of choice for such comparison because they are common in benthic aquatic habitats and accumulate substantial amounts of these compounds. Hexachlorobenzene, hexachlorocyclohexanes,4,4'-DDE,4,4'-DDT, polychlorobiphenyls, and PBDEs have been measured. The results show a nonselective enrichment of OCs and PBDEs from larvae to pupae. These concentration increases may result from the weight loss of pupae during metamorphosis as a consequence of mainly protein carbon respiration and lack of feeding. Despite the lack of change in total amount, the concentration increases from larvae to pupae are very relevant for the pollutant ingestion of the higher predators. The intakes of OCs and PBDEs by trout are between 2- and 5-fold higher per calorie gained when predating on pupae than on larvae. Since pollutant concentration, energy reward, predation susceptibility, and duration of life stage are very different between these two insect stages, and none of them is irrelevant for the incorporation of OCs or PBDEs to higher levels, bioaccumulation food-web models should distinguish between the two sources.     

Dyeing behaviour of relaxed and retractile components in high-bulk yarns

The evolution of dye absorption and fine structure variations due to heat treatment in high-bulk yarns was studied. Two acrylic fibres of different commercial origin were employed. Using these, the following yarns were spun: 100% relaxed (N), 100% retractile (R), 55:45 N/R and 45:55 N/R. The evolution of fibre microstructure induced by the dyeing process was studied through the differential solubility technique in mixtures of dimethylformamide and water.     

Paradigmatic De Novo GRIN1 Variants Recapitulate Pathophysiological Mechanisms Underlying GRIN1-Related Disorder Clinical Spectrum

Background: GRIN-related disorders (GRD), the so-called grinpathies, is a group of rare encephalopathies caused by mutations affecting GRIN genes (mostly GRIN1, GRIN2A and GRIN2B genes), which encode for the GluN subunit of the N-methyl D-aspartate (NMDA) type ionotropic glutamate receptors. A growing number of functional studies indicate that GRIN-encoded GluN1 subunit disturbances can be dichotomically classified into gain- and loss-of-function, although intermediate complex scenarios are often present. Methods: In this study, we aimed to delineate the structural and functional alterations of GRIN1 disease-associated variants, and their correlations with clinical symptoms in a Spanish cohort of 15 paediatric encephalopathy patients harbouring these variants. Results: Patients harbouring GRIN1 disease-associated variants have been clinically deeply-phenotyped. Further, using computational and in vitro approaches, we identified different critical checkpoints affecting GluN1 biogenesis (protein stability, subunit assembly and surface trafficking) and/or NMDAR biophysical properties, and their association with GRD clinical symptoms. Conclusions: Our findings show a strong correlation between GRIN1 variants-associated structural and functional outcomes. This structural-functional stratification provides relevant insights of genotype-phenotype association, contributing to future precision medicine of GRIN1-related encephalopathies.     

Toward a Novel Drug To Target the EGF–EGFR Interaction: Design of Metabolically Stable Bicyclic Peptides

In cancer, proliferation of malignant cells is driven by overactivation of growth‐signalling mechanisms, such as the epidermal growth factor receptor (EGFR) pathway. Despite its therapeutic relevance, the EGF–EGFR interaction has remained elusive to inhibition by synthetic molecules, mostly as a result of its large size and lack of binding pockets and cavities. Designed peptides, featuring cyclic motifs and other structural constraints, have the potential to modulate such challenging protein–protein interactions (PPIs). Herein, we present the structure‐based design of a series of bicyclic constrained peptides that mimic an interface domain of EGFR and inhibit the EGF–EGFR interaction by targeting the smaller partner (i.e., EGF). This design process was guided by the integrated use of in silico methods and biophysical techniques, such as NMR spectroscopy and surface acoustic wave. The best analogues were able to reduce selectively the viability of EGFR+ human cancer cells. In addition to their efficacy, these bicyclic peptides are endowed with exceptional stability and metabolic resistance—two features that make them suitable candidates for in vivo applications.     

The incorporation of quantum effects in enzyme kinetics modeling

We present an overview of new procedures for including quantum mechanical effects in enzyme kinetics. Quantum effects are included in three ways: (1) The electronic structure of the atoms in the catalytic center is treated quantum mechanically in order to calculate a realistic potential energy surface for the bond rearrangement process. (2) The discrete nature of quantum mechanical vibrational energies is incorporated in the treatment of nuclear motion for computing the potential of mean force. (3) Multidimensional tunneling contributions are included. These procedures are illustrated by applications to proton abstractions catalyzed by enolase and methylamine dehydrogenase and hydride-transfer reactions by alcohol dehydrogenase and xylose isomerase.