Delta2D and Proteomweaver: Performance evaluation of two different approaches for 2‐DE analysis

2‐DE is a fundamental technology used in proteomics research. However, despite its high capacity to simultaneously separate several proteins for subsequent identification and quantitative comparison studies, a drawback for this technique is its limited reproducibility, especially when comparing data from different laboratories. 2‐DE‐related variability can be broadly divided into two categories: experimental and post‐experimental. Experimental variability depends on physical and chemical parameters, whereas post‐experimental variability arises when gels are analyzed by different software packages, particularly when different workflows are followed. In this paper, we compared the analysis performance of two software packages, Delta2D and Proteomweaver, using both standard and experimental gel images. Using standard gel images, the false negative spot count was 50% lower, the false positive count was 77% lower, the true positive count was 19% higher and spot matching was 4% higher in Delta2D when compared to Proteomeweaver. Using experimental gel images, we found that the total amount of time taken to complete the analysis with Delta2D was 30% that of the time needed with Proteomweaver and required fewer user interventions. The differences between ease of use and workflow strategy of these programs is discussed.     

From domino to multicomponent: synthesis of dihydroisobenzofurans

A variety of substituted dihydroisobenzofurans can be easily synthesized in high yields by a one-pot three-component approach starting from o-bromoarylaldehydes, methanol, and terminal alkynes. The reaction occurs through an unprecedented cooperative palladium/base promoted coupling/addition/cyclization sequence.     

New 4-Aminoproline-Based Small Molecule Cyclopeptidomimetics as Potential Modulators of α4β1 Integrin

Integrin α₄β₁ belongs to the leukocyte integrin family and represents a therapeutic target of relevant interest given its primary role in mediating inflammation, autoimmune pathologies and cancer-related diseases. The focus of the present work is the design, synthesis and characterization of new peptidomimetic compounds that are potentially able to recognize α₄β₁ integrin and interfere with its function. To this aim, a collection of seven new cyclic peptidomimetics possessing both a 4-aminoproline (Amp) core scaffold grafted onto key α₄β₁-recognizing sequences and the (2-methylphenyl)ureido-phenylacetyl (MPUPA) appendage, was designed, with the support of molecular modeling studies. The new compounds were synthesized through SPPS procedures followed by in-solution cyclization maneuvers. The biological evaluation of the new cyclic ligands in cell adhesion assays on Jurkat cells revealed promising submicromolar agonist activity in one compound, namely, the c[Amp(MPUPA)Val-Asp-Leu] cyclopeptide. Further investigations will be necessary to complete the characterization of this class of compounds.     

Biodegradable Polymers from Renewable Sources. New Hemicellulose‐Based Hydrogels

Hemicellulose/poly(2‐hydroxyethyl methacrylate) (PHEMA)‐based hydrogels were prepared by the radical polymerization of HEMA with hemicellulose purposely modified with well‐defined amounts of methacrylic functions using a redox initiator system. Oligomeric hydrosoluble hemicelluloses produced from spruce chips were used for modification studies. The chemical modification of hemicellulose was performed with 2‐[(1‐imidazolyl)formyloxy]ethyl methacrylate as a modifying agent and triethylamine as the catalyst. The kinetics of the modification reaction was monitored by means of ¹H NMR spectroscopy. The resulting hydrogels obtained after a 30 min reaction were homogeneous, elastic, and transparent materials.     

Enzyme‐Mediated Preparation of (+)‐ and (−)‐β‐Irone and (+)‐ and (−)‐cis‐γ‐Irone from Irone alpha®

The (−)‐ and (+)‐β‐irones ((−)‐ and (+)‐ 2 , resp.), contaminated with ca. 7 – 9% of the (+)‐ and (−)‐trans‐α‐isomer, respectively, were obtained from racemic α‐irone via the 2,6‐trans‐epoxide (±)‐ 4 (Scheme 2). Relevant steps in the sequence were the LiAlH₄ reduction of the latter, to provide the diastereoisomeric‐4,5‐dihydro‐5‐hydroxy‐trans‐α‐irols (±)‐ 6 and (±)‐ 7 , resolved into the enantiomers by lipase‐PS‐mediated acetylation with vinyl acetate. The enantiomerically pure allylic acetate esters (+)‐ and (−)‐ 8 and (+)‐ and (−)‐ 9 , upon treatment with POCl₃/pyridine, were converted to the β‐irol acetate derivatives (+)‐ and (−)‐ 10 , and (+)‐ and (−)‐ 11 , respectively, eventually providing the desired ketones (+)‐ and (−)‐ 2 by base hydrolysis and MnO₂ oxidation. The 2,6‐cis‐epoxide (±)‐ 5 provided the 4,5‐dihydro‐4‐hydroxy‐cis‐α‐irols (±)‐ 13 and (±)‐ 14 in a 3 : 1 mixture with the isomeric 5‐hydroxy derivatives (±)‐ 15 and (±)‐ 16 on hydride treatment (Scheme 1). The POCl₃/pyridine treatment of the enantiomerically pure allylic acetate esters, obtained by enzymic resolution of (±)‐ 13 and (±)‐ 14 , provided enantiomerically pure cis‐α‐irol acetate esters, from which ketones (+)‐ and (−)‐ 22 were prepared (Scheme 4). The same materials were obtained from the (9S) alcohols (+)‐ 13 and (−)‐ 14 , treated first with MnO₂, then with POCl₃/pyridine (Scheme 4). Conversely, the dehydration with POCl₃/pyridine of the enantiomerically pure 2,6‐cis‐5‐hydroxy derivatives obtained from (±)‐ 15 and (±)‐ 16 gave rise to a mixture in which the γ‐irol acetates 25a and 25b and 26a and 26b prevailed over the α‐ and β‐isomers (Scheme 5). The (+)‐ and (−)‐cis‐γ‐irones ((+)‐ and (−)‐ 3 , resp.) were obtained from the latter mixture by a sequence involving as the key step the photochemical isomerization of the α‐double bond to the γ‐double bond. External panel olfactory evaluation assigned to (+)‐β‐irone ((+)‐ 2 ) and to (−)‐cis‐γ‐irone ((−)‐ 3 ) the strongest character and the possibility to be used as dry‐down note.     

Tomato and Olive Bioactive Compounds: A Natural Shield against the Cellular Effects Induced by β-Hexachlorocyclohexane-Activated Signaling Pathways

The β-isomer of hexachlorocyclohexane (β-HCH) is a globally widespread pollutant that embodies all the physicochemical characteristics of organochlorine pesticides, constituting an environmental risk factor for a wide range of noncommunicable diseases. Previous in vitro studies from our group disclosed the carcinogenic potential of β-HCH, which contributes to neoplastic transformation by means of multifaceted intracellular mechanisms. Considering the positive evidence regarding the protective role of natural bioactive compounds against pollution-induced toxicity, micronutrients from olive and tomato endowed with the capability of modulating β-HCH cellular targets were tested. For this purpose, the solution obtained from a patented food supplement (No. EP2851080A1), referred to as Tomato and Olive Bioactive Compounds (TOBC), was administered to the androgen-sensitive prostate cancer cells LNCaP and different biochemical and cellular assays were performed to evaluate its efficiency. TOBC shows a dose-dependent significant chemoprotection by contrasting β-HCH-induced intracellular responses such as STAT3 and AhR activation, disruption of AR signaling, antiapoptotic and proliferative activity, and increase in ROS production and DNA damage. These experimental outcomes identified TOBC as a suitable functional food to be included in a diet regimen aimed at defending cells from β-HCH negative effects, recommending the development of tailored enriched formulations for exposed individuals.     

Soluble Polystyrenes Functionalized by Triorgano[(1‐oxoalkyl)oxy]stannanes (=Triorganotin Carboxylates): Synthesis,Structure, and Anion‐Recognition Characteristics

Polystyrene copolymers of the type ( P −H)_1−x( P −(CH₂)_n−COOSnR₃)_x containing [(1‐oxoalkyl)oxy]triphenylstannane or tributyl[(1‐oxoalkyl)oxy]stannanes as side chains ( P −H=styrene; P −(CH₂)_n−COOSnR₃ =para‐substituted styrene‐like monomeric unit with R=Ph (x=0.1), Bu (x=0.5); n=2–4) were investigated. The tributyl[(1‐oxoalkyl)oxy]stannane copolymer was prepared by direct conversion of the corresponding copolymeric methyl esters with hexabutyldistannoxane. By contrast, the [(1‐oxoalkyl)oxy]triphenylstannane copolymer could be prepared only by a procedure involving two reaction steps consisting of a preliminary hydrolysis of the related methyl ester ( P −H)_1‐x( P −(CH₂)_n−COOMe)_x followed by functionalization of the corresponding poly(carboxylic acid) ( P −H)_1‐x( P −(CH_2n−COOH)_x with hydroxytriphenylstannane. Attempts to directly convert the methyl ester with hydroxytriphenylstannane or hexaphenyldistannoxane led to the formation of uncompletely functionalized product. The structure of the stannane‐functionalized polymers was investigated in solution and solid state by NMR, IR, and thermal analysis. The tributylstannane and triphenylstannane copolymers were assessed as chloride‐selective anion carriers in polymeric‐liquid‐membrane potentiometric ion‐selective electrodes.