Highly Concentrated Stabilized Hybrid Complexes of Hyaluronic Acid: Rheological and Biological Assessment of Compatibility with Adipose Tissue and Derived Stromal Cells towards Regenerative Medicine.

Cells and extracts derived from adipose tissue are gaining increasing attention not only in plastic surgery and for aesthetic purposes but also in regenerative medicine. The ability of hyaluronan (HA) to support human adipose stromal cell (hASC) viability and differentiation has been investigated. However, the compatibility of adipose tissue with HA-based formulation in terms of biophysical and rheological properties has not been fully addressed, although it is a key feature for tissue integration and in vivo performance. In this study, the biophysical and biochemical properties of highly concentrated (45 mg/mL) high/low-molecular-weight HA hybrid cooperative complex were assessed with a further focus on the potential application in adipose tissue augmentation/regeneration. Specifically, HA hybrid complex rheological behavior was observed in combination with different adipose tissue ratios, and hyaluronidase-catalyzed degradation was compared to that of a high-molecular-weight HA (HHA). Moreover, the HA hybrid complex's ability to induce in vitro hASCs differentiation towards adipose phenotype was evaluated in comparison to HHA, performing Oil Red O staining and analyzing gene/protein expression of PPAR-γ, adiponectin, and leptin. Both treatments supported hASCs differentiation, with the HA hybrid complex showing better results. These outcomes may open new frontiers in regenerative medicine, supporting the injection of highly concentrated hybrid formulations in fat compartments, eventually enhancing residing staminal cell differentiation and improving cell/growth factor persistence towards tissue regeneration districts.

Genomic basis for an informed conservation management of Pelophylax water frogs in Luxembourg.

Genetic identification methods have become increasingly important for species that are difficult to identify in the field. A case in point is Pelophylax water frogs. While their morphological determination is highly complex, they include species protected under EU law and some that are classified as invasive. Additionally, genetic data can provide insights into their complex breeding systems, which may or may not involve the reproductive dependency of one species on another. Here, we generate baseline data for water frog monitoring in Luxembourg. We applied a countrywide sampling approach and used SNPs generated by ddRAD sequencing to identify individuals and infer the breeding systems present in the country. We found Pelophylax lessonae and P. kl. esculentus throughout Luxembourg, mostly living in syntopy. In general, a reproductive dependency of P. kl. esculentus on P. lessonae (L-E system) was revealed. Besides this general system, we detected triploid P. kl. esculentus in six ponds. This indicates a modified L-E system with reproductive dependency of the triploids on the diploid P. kl. esculentus. The invasive P. cf. bedriagae was detected in three ponds in southern Luxembourg, with evidence for hybridization with native water frogs. In addition to the ddRAD data, we tested a simple genetic method for future monitoring based on the MND1 marker. It showed in almost all cases, an identical species identification as the ddRAD data and was successfully applied to DNA extracts from mouth swabs. Combining this method with our baseline data will enable informed choices for the protection of native water frog species in Luxembourg.

A synergetic and sensitive physostigmine pesticide sensor using copper complex of 3D zinc (II) phthalocyanine-SWCNT hybrid material.

2,3,9,10,16,17,23,24-Octakis (4-methyl-2,6-bis((prop-2-yn-1-yloxy)methyl)phenoxy) phthalocyaninato zinc(II) (Pc) bearing sixteen terminal ethynyl groups was synthesized and attached to SWCNT (Single-walled carbon nanotube) covalently to obtain three dimensional porous hybrid material (SWCNT-Pc 3D) and its copper complex (Cu-SWCNT-Pc 3D). The structural characterization and electrochemical sensor features of the Cu-SWCNT-Pc hybrid towards to physostigmine pesticide were performed. A fast, direct and suitable determination method for physostigmine detection was offered. The designed sensor, Cu-SWCNT-Pc 3D/GCE (glassy carbon electrode) shows sensitivity ca 1.8, 4.3 and 2.8 times more than that of SWCNT/GCE, SWCNT-Pc-noncovalent/GCE and SWCNT-Pc 3D/GCE in terms of peak heights while bare and Pc/GCE had almost no voltammetric response to 2 µM physostigmine in PBS at a pH of 7.0. The limit of detection and quantification of physostigmine determination with Cu-SWCNT-Pc 3D/GCE were found to be 53 and 177 nM in the range of 0.1-4.8 µM, respectively. This study demonstrated that the modification of the GCE with Cu-SWCNT-Pc 3D as an electrochemical sensor was acted as catalytic role toward physostigmine presence of other interfering pesticides as high sensitivity and selectivity. The electrochemical determination of physostigmine in real samples was performed under the optimized conditions, also accuracy of the electrochemical determination method was evaluated with HPLC as a standard determination method.

CRISPR/Cas9-Mediated Multi-Allelic Gene Targeting in Sugarcane Confers Herbicide Tolerance.

Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n = 100 - 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple alleles via template-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions.

A new organic-inorganic compound, ethyl-enedi-ammonium hexa-chlorido-stannate(IV) p-anisaldehyde disolvate.

The asymmetric unit of the title organic-inorganic hybrid complex [systematic name: ethane-1,2-diaminium hexa-chlorido-stannate(IV)-4-meth-oxy-benz-alde-hyde (1/2)], (C2H10N2)[SnCl6]·2C8H8O2, contains one half of an ethyl-enedi-ammonium cation, one half of an [SnCl6]2- anion and one p-anisaldehyde mol-ecule. Both the organic cation and the quasi-regular octa-hedral inorganic anion are located about inversion centres. The organic cations and [SnCl6]2- anions lie in layers parallel to the ac plane with p-anisaldehyde mol-ecules occupying the space between the layers. A network of classical N-H⋯Cl and N-H⋯O hydrogen bonds exists between the ethyl-enedi-ammonium cations and the [SnCl6]2- anions and p-anisaldehyde mol-ecules. These inter-actions, together with non-classical C-H⋯O inter-actions between the ethyl-enedi-ammonium cations and the p-anisaldehyde mol-ecules, serve to hold the structure together. The crystal studied was refined as a two-component twin.

Oxygen-releasing manganese clay hybrid complex triggers p53-mediated cancer cell death in hypoxia.

Hypoxia in tumor microenvironment is responsible for resistance to conventional modes of cancer therapeutics. A manganese-clay hybrid compound MHC was shown to generate molecular oxygen in aqueous solution. In this study we have shown that MHC, in hypoxia, causes cancer cell death, through release of molecular oxygen and via p53-dependent apoptosis. MHC treatment of cells results in depletion of mitochondrial membrane potential and inhibition of ROS production, in a cell-specific manner. In hypoxia, the oxygen from MHC releases cells from S-phase arrest thus causing p53-dependent apoptosis. The induction of apoptosis by MHC is higher in p53 Wt/Wt cells when it is compared with p53 Mt/Mt cells. The released oxygen from MHC triggers apoptosis via p53 activation through its enhanced homo-oligomerization, post-translational modifications and nuclear localization. Thus MHC as a cellular oxygen-releasing compound has high potential as a drug for hypoxic tumor regression.

A hybrid siRNA delivery complex for enhanced brain penetration and precise amyloid plaque targeting in Alzheimer's disease mice.

To realize the therapeutic potential of gene drugs for Alzheimer's disease (AD), non-invasive, tissue-specific and efficient delivery technologies must be developed. Here, a hybrid system for amyloid plaques targeted siRNA delivery was formed by PEGylated Poly(2-(N,N-dimethylamino) ethyl methacrylate) (PEG-PDMAEMA) conjugated with two d-peptides, a CGN for brain penetration and a QSH for ß-amyloid binding. The hybrid complex CQ/siRNA, composed of 25% MPEG-PDMAEMA, 50% CGN-PEG-PDMAEMA and 25% QSH-PEG-PDMAEMA, showed negligible cytotoxicity and could protect siRNA from enzyme degradation. Being taken up by neuron cells, the complexes could escape from lysosomes, release siRNA in the cytoplasm and thus producing effective gene silence (down-regulated protein level to 18.5%). After intravenous injection, CQ/siRNA penetrated into the brain in an intact form and located around the plaques in transgenic AD mice. The precisely amyloid plaques delivery resulted in increased therapeutic activities, which was demonstrated by the strong mRNA (36.4%) knockdown of BACE1 (a therapeutic target of AD), the less yield of enzyme-digested products sAPPß (-42.6%), as well as the better neurons protection than the single component complexes. In conclusion, the hybrid complex could efficiently and precisely deliver an siRNA to the AD lesion and might be a potential candidate for gene therapy for AD. STATEMENT OF SIGNIFICANCE: The gene delivery system achieving high brain penetration and lesion region accumulation was first applied to treat AD, and the preparation exhibited a significantly better neuroprotective effect than that modified with a single ligand. The intracellular process of which the complexes escape from lysosomes and release the siRNA in cytoplasm was revealed. The brain targeting and amyloid plaque binding ability of the complex were systemic evaluated, and the in vivo co-location experiments provided a direct evidence of the precise delivery of the siRNA to the amyloid plaques. One of the targeting ligands, CGN, which was a retro-inverso modified peptide to achieve better affinity to the BBB, was first applied to the brain targeting system.

Synthesis and structural studies of a new complex of catena-poly[p-anisidinium [[diiodidobismu-thate(III)]-di-µ-iodido] dihydrate].

A new organic-inorganic hybrid material, {(C7H10NO)[BiI4]·2H2O} n , has been synthesized by slow evaporation of an aqueous solution at room temperature. The anionic sublattice of the crystal is built up by [BiI6] octa-hedra sharing edges. The resulting zigzag chains extend along the a-axis direction and are arranged in a distorted hexagonal rod packing. The p-anisidinium cations and the water mol-ecules are located in the voids of the anionic sublattice. The cations are linked to each other through N-H⋯O hydrogen bonds with the water mol-ecules, and also through weaker N-H⋯I inter-actions to the anionic inorganic layers.

Influence of amodiaquine on the antimalarial activity of ellagic acid: crystallographic and biological studies.

In the search for new antimalarial drugs, design of hybrid molecules is recommended to improve biological activity and to decrease the risk of parasite resistance development. Ellagic acid, as an inhibitor of Plasmodium glutathione, presents an original mode of action and thus appears as a promising antiplasmodial compound. A new complex (AQ-EA) consisting of the well-known antimalarial drug, amodiaquine, and ellagic acid was obtained. The studied crystal structure of AQ-EA showed that the triclinic centrosymmetrical unit cell of the crystal contains two molecules of amodiaquine (AQ) and two symmetrically independent molecules of ellagic acid (EA). The packing of the molecules in the crystal is dominated by hydrogen bonds between AQ and EA. The antiplasmodial activity of the hybrid complex AQ-EA was also determined and compared with the values of IC50 for AQ and EA separately. Potentiation assays between both molecules were conducted to understand the pharmacological interactions between AQ and EA against Plasmodium falciparum in vitro. The hybrid complex AQ-EA (IC50 of 47 nm) showed improved antiplasmodial activity in comparison with EA alone.

A one-dimensional ladder-like six-coordinate cadmium polymer: catena-poly[bis[(S)-2-methylpiperazine-1,4-diium] [bis[trichloridocadmium(II)]-di-µ3-chlorido]].

The crystal structure of the title compound {(C5H14N2)2[Cd2Cl8]}n, (I), consists of hydrogen-bonded 2-methylpiperazinediium (H2MPPA(2+)) cations in the presence of one-dimensional polymeric {[CdCl3(µ3-Cl)](2-)}n anions. The Cd(II) centres are hexacoordinated by three terminal chlorides and three bridging chlorides and have a slightly distorted octahedral CdCl3(µ3-Cl)3 arrangement. The alternating CdCl6 octahedra form four-membered Cd2Cl2 rings by the sharing of neighbouring Cd-Cl edges to give rise to extended one-dimensional ladder-like chains parallel to the b axis, with a Cd···Cd distance of 4.094 (2) Å and a Cd···Cd···Cd angle of 91.264 (8)°. The H2MPPA(2+) cations crosslink the [CdCl3(µ3-Cl)]n chains by the formation of two N-H···Cl hydrogen bonds to each chain, giving rise to one-dimensional ladder-like H2MPPA(2+)-Cl2 hydrogen-bonded chains [graph set R4(2)(14)]. The [CdCl3(µ3-Cl)]n chains are interwoven with the H2MPPA(2+)-Cl2 hydrogen-bonded chains, giving rise to a three-dimensional supramolecular network.