Effects of feeding elevated concentrations of monounsaturated fatty acids and vitamin E to swine on characteristics of dry cured hams

Thirty Large White×Great York gilts were fed six experimental diets containing three levels of poly and monounsaturated fatty acids. Within each dietary fat treatment, one group was fed a basal level of vitamin E (20 mg α-tocopheryl acetate/kg diet) and the other group received a supplemented level (200 mg α-tocopheryl acetate/kg diet). Concentration of α-tocopherol was significantly higher in hams from pigs fed supplemented dietary levels of vitamin E (P<0.0001), but no significant effect of dietary fat was observed. Dietary vitamin E supplementation reduced the concentration of thiobarbituric acid reactive substances after 9 days storage of sliced samples (P<0.0001), while dietary fat source showed no significant effect. Significantly lower oxidation was observed in ham homogenates from pigs fed higher concentrations of monounsaturated fatty acids after 120 min of incubation under pro-oxidant conditions (P=0.013). No effect of dietary treatment was observed in ham volatile aldehyde profile. No significant effect of dietary vitamin E was observed on surface redness during storage, but a significant effect was observed for luminosity after 7 days of storage (P=0.033). Hams from pigs fed diets enriched in monounsaturated fatty acid showed higher ‘a’ values (P=0.040) in stored sliced samples. Sensory evaluation revealed a significant effect of dietary vitamin E on redness of ham slices (P<0.001). Dietary supplementation with vitamin E also produced a significantly higher odour and flavour intensity (P=0.006 and P=0.01 respectively). Dry cured ham samples from pigs fed higher amounts of monounsaturated fatty acids showed a significantly higher consistency fat than those from pigs fed polyunsaturated fatty acids.     

Structural autonomy of a β-hairpin peptide derived from the pneumococcal choline-binding protein LytA

The cell wall of Streptococcus pneumoniae and several other micro-organisms is decorated with a number of the so-called choline-binding proteins (CBPs) that recognise the choline residues in the bacterial surface by means of highly conserved, concatenated 20-aa sequences termed choline-binding repeats (CBRs), that are composed of a loop and a β-hairpin structure. In this work, we have investigated the ability to fold in aqueous solution of a 14-aa peptide (LytA???????[wt]) and a single derivative of it, LytA???????[ND], corresponding to one of the six β-hairpins of the LytA pneumococcal amidase. Intrinsic fluorescence and circular dichroism spectroscopical measurements showed that both peptides spontaneously acquire a non-random conformation which is also able to bind the natural ligand choline. Furthermore, nuclear magnetic resonance techniques allowed the calculation of the structure of the LytA???????[ND] peptide, which displayed a β-hairpin conformation highly similar to that found within the full-length C-LytA module. These results provide a structural basis for the modular organisation of CBPs and suggest the use of CBRs as new templates for the design of stable β-hairpins.     

Analysis of the Effect of Al on the Static Softening Kinetics of C-Mn Steels Using a Physically Based Model

The effect of Al addition on the static softening behavior of C-Mn steels was investigated. Double-hit torsion tests were performed at different deformation temperatures ranging from 1198 K to 1338 K (925 °C to 1065 °C) with pass strains of ε = 0.2 and 0.35. It was found that solute Al produced a significant delay on the static softening kinetics. Additionally, at the lowest temperatures [1198 K to 1238 K (925 °C to 965 °C)] and highest Al level (2 wt pct), austenite to ferrite phase transformation was found to be concurrent with softening, leading this to higher softening retardation. The softening kinetics of the steels investigated were analyzed using a physically based model which couples recovery and recrystallization mechanisms. The main parameters of the model were identified for the present alloys. An expression for the grain boundary mobility of the base C-Mn steel was derived and the retarding effect of Al in solid solution on the static recrystallization kinetics was introduced in the model. Reasonable agreement was obtained between model and experimental results for a variety of deformation conditions.     

TGF-β Signalling Mediates the Anti-Inflammatory Activity of Enamel Matrix Derivative In Vitro

Enamel matrix derivative (EMD) prepared from extracted porcine fetal tooth material can support the regrow of periodontal tissues. Previous findings suggest that EMD has anti-inflammatory properties and TGF-β activity in vitro. However, the anti-inflammatory activity of EMD is mediated via TGF-β has not been considered. To this aim, we first established a bioassay to confirm the anti-inflammatory activity of EMD. The bioassay was based on the RAW 264.7 macrophage cell line and proven with primary macrophages where EMD significantly reduced the forced expression of IL-6. We then confirmed the presence of TGF-β1 in EMD by immunoassay and by provoking the Smad2/3 nuclear translocation in RAW 264.7 macrophages. Next, we took advantage of the TGF-β receptor type I kinase-inhibitor SB431542 to block the respective signalling pathway. SB431542 reversed the anti-inflammatory activity of EMD and TGF-β in a bioassay when IL-6 and CXCL2 expression was driven by the LPS stimulation of RAW 264.7 macrophages. This central observation was supported by showing that SB431542 reversed the anti-inflammatory activity of EMD using IL-1β and TNF-α-stimulated ST2 bone marrow stromal cells. Together, these findings implicate that the TGF-β activity mediates at least part of the anti-inflammatory activity of EMD in vitro.     

Enantiocontrolled Preparation of ϒ-Substituted Cyclohexenones: Synthesis and Kinase Activity Assays of Cyclopropyl-Fused Cyclohexane Nucleosides

The enantioselective preparation of the two isomers of 4-hydroxy-2-cyclohexanone derivatives 1a,b was achieved, starting from a common cyclohexenone, through asymmetric transfer hydrogenation (ATH) reactions using bifunctional ruthenium catalysts. From these versatile intermediates, a stereoselective route to a cytosine analogue built on a bicyclo [4.1.0]heptane scaffold is described. Nucleoside kinase activity assays with this cyclopropyl-fused cyclohexane nucleoside, together with other related nucleosides (2a–e), were performed, showing that thymine- and guanine- containing compounds have affinity for herpes simplex virus Type 1 (HSV-1) thymidine kinase (TK) but not for human cytosolic TK-1, thus pointing to their selectivity for herpetic TKs but not cellular TKs.     

Molecular Basis of the Schuurs–Hoeijmakers Syndrome: What We Know about the Gene and the PACS-1 Protein and Novel Therapeutic Approaches

The Schuurs–Hoeijmakers syndrome (SHMS) or PACS1 Neurodevelopment Disorder (PACS1-NDD) is a rare autosomal dominant disease caused by mutations in the PACS1 gene. To date, only 87 patients have been reported and, surprisingly, most of them carry the same variant (c.607C>T; p.R203W). The most relevant clinical features of the syndrome include neurodevelopment delay, seizures or a recognizable facial phenotype. Moreover, some of these characteristics overlap with other syndromes, such as the PACS2 or Wdr37 syndromes. The encoded protein phosphofurin acid cluster sorting 1 (PACS-1) is able to bind to different client proteins and direct them to their subcellular final locations. Therefore, although its main function is protein trafficking, it could perform other roles related to its client proteins. In patients with PACS1-NDD, a gain-of-function or a dominant negative mechanism for the mutated protein has been suggested. This, together with the fact that most of the patients carry the same genetic variant, makes it a good candidate for novel therapeutic approaches directed to decreasing the toxic effect of the mutated protein. Some of these strategies include the use of antisense oligonucleotides (ASOs) or targeting of its client proteins.     

Functional Delineation of a Protein–Membrane Interaction Hotspot Site on the HIV-1 Neutralizing Antibody 10E8

Antibody engagement with the membrane-proximal external region (MPER) of the envelope glycoprotein (Env) of HIV-1 constitutes a distinctive molecular recognition phenomenon, the full appreciation of which is crucial for understanding the mechanisms that underlie the broad neutralization of the virus. Recognition of the HIV-1 Env antigen seems to depend on two specific features developed by antibodies with MPER specificity: (i) a large cavity at the antigen-binding site that holds the epitope amphipathic helix; and (ii) a membrane-accommodating Fab surface that engages with viral phospholipids. Thus, besides the main Fab–peptide interaction, molecular recognition of MPER depends on semi-specific (electrostatic and hydrophobic) interactions with membranes and, reportedly, on specific binding to the phospholipid head groups. Here, based on available cryo-EM structures of Fab–Env complexes of the anti-MPER antibody 10E8, we sought to delineate the functional antibody–membrane interface using as the defining criterion the neutralization potency and binding affinity improvements induced by Arg substitutions. This rational, Arg-based mutagenesis strategy revealed the position-dependent contribution of electrostatic interactions upon inclusion of Arg-s at the CDR1, CDR2 or FR3 of the Fab light chain. Moreover, the contribution of the most effective Arg-s increased the potency enhancement induced by inclusion of a hydrophobic-at-interface Phe at position 100c of the heavy chain CDR3. In combination, the potency and affinity improvements by Arg residues delineated a protein–membrane interaction site, whose surface and position support a possible mechanism of action for 10E8-induced neutralization. Functional delineation of membrane-interacting patches could open new lines of research to optimize antibodies of therapeutic interest that target integral membrane epitopes.