Direct investigation of the vectorization properties of amphiphilic cyclodextrins in phospholipid films

Recently, new cyclodextrin derivatives were synthesized and shown to exhibit strong amphiphilic properties. In this paper, we study the action of these new amphiphilic cyclodextrins on phospholipids. Mixed phospholipid/cyclodextrin derivative films were prepared and studied using X-ray reflectivity for various phospholipid/cyclodextrin ratios. A molar ratio of 3 provides a highly stable film the molecular structure of which has been investigated in detail. The cholesterol tail of the cyclodextrin molecule was found to be anchored into the phospholipid film. The cyclodextrin moieties exposed to the aqueous medium are prone to the addition of the guest molecule Dosulepin, making them of high interest for drug delivery. For this purpose and as an example of a potential application, this cyclodextrin molecular carrier property is also addressed to this complex film architecture.     

A DNA aptamer as a new target-specific chiral selector for HPLC

In this paper, a DNA aptamer, known to bind stereospecifically the D-enantiomer of an oligopeptide, i.e., arginine-vasopressin, was immobilized on a chromatographic support. The influence of various parameters (such as column temperature, eluent pH, and salt concentration) on the L- and D-peptide retention was investigated in order to provide information about the binding mechanism and then to define the utilization conditions of the aptamer column. The results suggest that dehydration at the binding interface, charge-charge interactions, and adaptive conformational transitions contribute to the specific D-peptide-aptamer complex formation. A very significant enantioselectivity was obtained in the optimal binding conditions, the D-peptide being strongly retained by the column while the L-peptide eluted in the void volume. A rapid baseline separation of peptide enantiomers was also achieved by modulating the elution conditions. Furthermore, it was established that the aptamer column was stable during an extended period of time. This work indicates that DNA aptamers, specifically selected against an enantiomer, could soon become very attractive as new target-specific chiral selectors for HPLC.     

Chemical modification of the adeno-associated virus capsid to improve gene delivery

Gene delivery vectors based on adeno-associated virus (AAV) are highly promising due to several desirable features of this parent virus, including a lack of pathogenicity, efficient infection of dividing and non-dividing cells and sustained maintenance of the viral genome. However, the conclusion from clinical data using these vectors is that there is a need to develop new AAVs with a higher transduction efficiency and specificity for relevant target tissues. To overcome these limitations, we chemically modified the surface of the capsid of AAV vectors. These modifications were achieved by chemical coupling of a ligand by the formation of a thiourea functionality between the amino group of the capsid proteins and the reactive isothiocyanate motif incorporated into the ligand. This strategy does not require genetic engineering of the capsid sequence. The proof of concept was first evidenced using a fluorophore (FITC). Next, we coupled the N-acetylgalactosamine ligand onto the surface of the AAV capsid for asialoglycoprotein receptor-mediated hepatocyte-targeted delivery. Chemically-modified capsids also showed reduced interactions with neutralizing antibodies. Taken together, our findings reveal the possibility of creating a specific engineered platform for targeting AAVs via chemical coupling.

Bioconjugated AAV vectors, achieved by coupling of ligands on amino groups of the capsid, are of great interest for gene delivery. Chemical modifications can be used to enhance cell tropism and to decrease interactions with neutralizing antibodies.     

Optimal focusing by spatio-temporal inverse filter. II. Experiments. Application to focusing through absorbing and reverberating media

To focus ultrasonic waves in an unknown heterogeneous medium using a phased array, one has to calculate the optimal set of signals to be applied on the transducers of the array. (In most applications of ultrasound, medical imaging, medical therapy, nondestructive testing, the first step consists of focusing a broadband ultrasound beam deeply inside the medium to be investigated.) Focusing in a homogeneous medium simply requires to compensate for the varying focus-array elements geometrical distances. Nevertheless, heterogeneities in the medium, in terms of speed of sound, density, or absorption, may strongly degrade the focusing. Different techniques have been developed in order to correct such aberrations induced by heterogeneous media (time reversal, speckle brightness, for example). In the companion to this paper, a new broadband focusing technique was investigated: the spatio-temporal inverse filter. Experimental results obtained in various media, such as reverberating and absorbing media, are presented here. In particular, intraplate echoes suppression and high-quality focusing through a human skull, as well as hyper-resolution in a reverberating medium, will be shown. It is important to notice that all these experiments were performed with fully programmable multichannel electronics whose use is required to fully exploit the spatio-temporal technique.     

Limits of Near‐Coloring of Sparse Graphs

Let be nonnegative integers. A graph G is ‐colorable if its vertex set can be partitioned into sets such that the graph induced by has maximum degree at most d for , while the graph induced by is an edgeless graph for . In this article, we give two real‐valued functions and such that any graph with maximum average degree at most is ‐colorable, and there exist non‐‐colorable graphs with average degree at most . Both these functions converge (from below) to when d tends to infinity. This implies that allowing a color to be d‐improper (i.e., of type ) even for a large degree d increases the maximum average degree that guarantees the existence of a valid coloring only by 1. Using a color of type (even with a very large degree d) is somehow less powerful than using two colors of type (two stable sets).     

The Prins reaction using ketones: rationalization and application toward the synthesis of the portentol skeleton

We report a TMSI-promoted Prins cyclization reaction with ketones as carbonyl partners to prepare polysubstituted chiral spirotetrahydropyrans. In the presence of racemic 2-methylcyclohexanone a dynamic kinetic resolution occurred affording one stereoisomer. The observed enantiospecificity has been rationalized by DFT calculation.     

On two variations of identifying codes

Identifying codes have been introduced in 1998 to model fault detection in multiprocessor systems. In this paper, we introduce two variations of identifying codes: weak codes and light codes. They correspond to fault detection by successive rounds. We give exact bounds for those two definitions for the family of cycles.     

Toward bioinspired galectin mimetics: identification of ligand-contacting peptides by proteolytic-excision mass spectrometry

Clinically relevant bioactivities of human galectins (adhesion/growth-regulatory galactoside-specific lectins) inspired the design of peptides as new tools to elicit favorable effects (e.g., in growth control) or block harmful binding (e.g., in tissue invasion). To obtain the bioinspired lead compounds, we combined a proteolytic fragmentation approach without/with ligand contact (excision) with mass spectrometric identification of affinity-bound protein fragments, using galectin-1 and -3 as models. Two peptides from the carbohydrate recognition domains were obtained in each case in experimental series rigorously controlled for specificity, and the [157-162] peptide of galectin-3 proved to be active in blocking lectin binding to a neoglycoprotein and to tumor cell surfaces. This approach affords peptide sequences for structural optimization and intrafamily/phylogenetic galectin comparison at the binding-site level with a minimal requirement of protein quantity, and it is even amenable to mixtures.     

Maturation of the [FeFe]-Hydrogenase: Direct Transfer of the (κ3-cysteinate)FeII(CN)(CO)2 Complex B from HydG to HydE

[FeFe]-hydrogenases efficiently catalyze the reversible oxidation of molecular hydrogen. Their prowess stems from the intricate H-cluster, combining a [Fe₄S₄] center with a binuclear iron center ([2Fe]_H). In the latter, each iron atom is coordinated by a CO and CN ligand, connected by a CO and an azadithiolate ligand. The synthesis of this active site involves a unique multiprotein assembly, featuring radical SAM proteins HydG and HydE. HydG initiates the transformation of L-tyrosine into cyanide and carbon monoxide to generate complex B, which is subsequently transferred to HydE to continue the biosynthesis of the [2Fe]_H-subcluster. Due to its instability, complex B isolation for structural or spectroscopic characterization has been elusive thus far. Nevertheless, the use of a biomimetic analogue of complex B allowed circumvention of the need for the HydG protein during in vitro functional investigations, implying a similar structure for complex B. Herein, we used the HydE protein as a nanocage to encapsulate and stabilize the complex B product generated by HydG. Using X-ray crystallography, we successfully determined its structure at 1.3 Å resolution. Furthermore, we demonstrated that complex B is directly transferred from HydG to HydE, thus not being released into the solution post-synthesis, highlighting a transient interaction between the two proteins.