Pectic Galactan Polysaccharide-Based Gene Delivery System for Targeting Neuroinflammation

Treating neuroinflammation-related injuries and disorders through manipulation of neuroinflammation functions is being heralded as a new therapeutic strategy. In this study, a novel pectic galactan (PG) polysaccharide based gene therapy approach is developed for targeting reactive gliosis in neuroinflammation. Galectin-3 (Gal-3) is a cell protein with a high affinity to β-galactoside sugars and is highly expressed in reactive gliosis. Since PG carries galactans, it can target reactive gliosis via specific carbohydrate interaction between galactan and Gal-3 on the cell membrane, and therefore can be utilized as a carrier for delivering genes to these cells. The carrier is synthesized by modifying quaternary ammonium groups on the PG. The resulting quaternized PG (QPG) is found to form complexes with plasmid DNA with a mean diameter of 100 nm and have the characteristics required for targeted gene therapy. The complexes efficiently condense large amounts of plasmid per particle and successfully bind to Gal-3. The in vivo study shows that the complexes are biocompatible and safe for administration and can selectively transfect reactive glial cells of an induced cortical lesion. The results confirm that this PG-based delivery system is a promising platform for targeting Gal-3 overexpressing neuroinflammation cells for treating neuroinflammation-related injuries and neurodegenerative diseases.     

Molecular Formation upon Ionization of van der Waals Clusters and Implication to Astrochemistry

The presence and formation of a large variety of organic molecules in the interstellar medium is evident from both astronomical data of absorption and emission bands at different regions of the spectrum. Specifically, polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in the interstellar medium (ISM). The mechanism for their formation in ISM‘s low temperature environment is, as of yet, a mystery nonetheless. Understanding the mechanism of formation of complex molecules such as PAHs and nitrogen-based PAHs (PANH) in the ISM is a long-standing challenge which has been drawing a growing attention for the past several decades. In this review we wish to emphasize two things: Firstly, the essential role quantum chemistry can play in the study of astrochemical reactions. Secondly, we wish to demonstrate that said variety of possibilities for chemical reaction, starting upon ionization of van der Waals clusters. The potential for different chemical reactions to occur within a cluster environment arises from the fact that such processes can take place at low temperatures as the systems pose large amounts of energy upon ionization. Moreover, the spectator molecules in the cluster can provide a dissipation route for energy by detachment from the cluster, thus the system can stabilize efficiently even at low densities. The spectator molecules can also change the potential energy surface, by which it will pose a catalytic effect for certain reactions. We will demonstrate this by presenting Ab Initio Molecular Dynamic results on ionization of small acetylene clusters.     

Low-Frequency Ultrasound Effects on Intracellular Barriers in Nonviral Gene Delivery Processes

Gene therapy, the expression in cells of genetic material with therapeutic activity, has emerged as a promising approach for the treatment or prevention of human diseases. At the present time, major somatic gene-transfer approaches employ either viral or nonviral vectors. Nonviral vectors are less efficient at introducing and maintaining foreign gene expression, but have the profound advantage of being nonpathogenic and nonimmunogenic. In this study, we aimed to develop an efficient nonviral gene delivery system in which low-frequency ultrasound (LFUS) was applied to enhance gene expression of polyplexes formed with poly(2-dimethylaminoethyl methacrylate) and plasmid encoding for green fluorescent protein. Ultrasound (US), and in particular LFUS, can cause temporary membrane permeabilization and thereby enhance drug and gene entrance into viable cells. We evaluated possible additional favorable effects of LFUS on the polyplex transfection process, such as overcoming intracellular barriers. We found that pDMAEMA protected the plasmid DNA from ultrasonic degradation. Atomic force microscopy analysis also confirmed that the LFUS did not change the polyplexes’ morphology. We also attained an insight into the structure of polyplexes during LFUS exposure and found that LFUS induced a temporary partial detachment between the polymer chains and the plasmid. In addition, LFUS application on ovarian carcinoma cells transfected with the polyplexes induced a 27% enhancement in transfection efficiency. Based on these results, we propose that LFUS enhances the decomplexation of the polyplexes, and therefore, can be used to optimize transfection efficiency.     

Nonadiabatic vibronic dynamics as a tool. From surface nanochemistry to coherently driven molecular machines

Resonances are ubiquitous in molecular heterojunctions and in scanning tunneling microscopy (STM) experiments. In the former environment, resonance tunneling is essential for favorable wire-length-dependence of the conductance and is often the mechanism underlying conductance enhancement through application of a gate voltage. In the latter environment, resonance tunneling has served to develop a powerful vibrational spectroscopy. Resonance conductance is often strongly nonadiabatic; in the course of the tunneling event, electron energy is channelled into vibrational modes and triggers molecular dynamics. The qualitative physics underlying current-driven, resonance-mediated dynamics in molecular electronics is very simple, and is familiar from related phenomena such as gas phase electron-molecule scattering and photochemistry on conducting surfaces. Equilibrium displacement between the initial and resonant states translates into vibronic coupling in the language of the Marcus theory of electron transfer; it produces a nonstationary superposition in the nuclear subspace that evolves during the resonance lifetime. Upon relaxation the system is internally excited and interesting dynamics is likely to ensue. While the underlying physics is very general, the single-molecule STM and molecular heterojunction environments open unique and exciting opportunities. The former introduces the possibility of determining resonance lifetimes through fit of experimental voltage dependencies to a quantum mechanical theory. The latter introduces the possibility of developing coherently driven molecular machines, a new form of nanolithography, and a new means of manipulating the conductivity of molecular-scale devices. We briefly review the theory of current-driven dynamics in molecular-scale devices, discuss the results of ongoing research on surface nanochemistry and molecular machines, and sketch a variety of potential applications.     

The Non-Erythropoietic EPO Analogue Cibinetide Inhibits Osteoclastogenesis In Vitro and Increases Bone Mineral Density in Mice

The two erythropoietin (EPO) receptor forms mediate different cellular responses to erythropoietin. While hematopoiesis is mediated via the homodimeric EPO receptor (EPOR), tissue protection is conferred via a heteromer composed of EPOR and CD131. In the skeletal system, EPO stimulates osteoclast precursors and induces bone loss. However, the underlying molecular mechanisms are still elusive. Here, we evaluated the role of the heteromeric complex in bone metabolism in vivo and in vitro by using Cibinetide (CIB), a non-erythropoietic EPO analogue that exclusively binds the heteromeric receptor. CIB is administered either alone or in combination with EPO. One month of CIB treatment significantly increased the cortical (~5.8%) and trabecular (~5.2%) bone mineral density in C57BL/6J WT female mice. Similarly, administration of CIB for five consecutive days to female mice that concurrently received EPO on days one and four, reduced the number of osteoclast progenitors, defined by flow cytometry as Lin⁻CD11b⁻Ly6C^hi CD115⁺, by 42.8% compared to treatment with EPO alone. In addition, CIB alone or in combination with EPO inhibited osteoclastogenesis in vitro. Our findings introduce CIB either as a stand-alone treatment, or in combination with EPO, as an appealing candidate for the treatment of the bone loss that accompanies EPO treatment.     

Myo-Inositol Limits Kainic Acid-Induced Epileptogenesis in Rats

Epilepsy is a severe neurological disease characterized by spontaneous recurrent seizures (SRS). A complex pathophysiological process referred to as epileptogenesis transforms a normal brain into an epileptic one. Prevention of epileptogenesis is a subject of intensive research. Currently, there are no clinically approved drugs that can act as preventive medication. Our previous studies have revealed highly promising antiepileptogenic properties of a compound–myo-inositol (MI) and the present research broadens previous results and demonstrates the long-term disease-modifying effect of this drug, as well as the amelioration of cognitive comorbidities. For the first time, we show that long-term treatment with MI: (i) decreases the frequency and duration of electrographic SRS in the hippocampus; (ii) has an ameliorating effect on spatial learning and memory deficit associated with epileptogenesis, and (iii) attenuates cell loss in the hippocampus. MI treatment also alters the expression of the glial fibrillary acidic protein, LRRC8A subunit of volume-regulated anion channels, and protein tyrosine phosphatase receptor type R, all expected to counteract the epileptogenesis. All these effects are still present even 4 weeks after MI treatment ceased. This suggests that MI may exert multiple actions on various epileptogenesis-associated changes in the brain and, therefore, could be considered as a candidate target for prevention of epileptogenesis.     

TENT4A Non-Canonical Poly(A) Polymerase Regulates DNA-Damage Tolerance via Multiple Pathways That Are Mutated in Endometrial Cancer

TENT4A (PAPD7) is a non-canonical poly(A) polymerase, of which little is known. Here, we show that TENT4A regulates multiple biological pathways and focuses on its multilayer regulation of translesion DNA synthesis (TLS), in which error-prone DNA polymerases bypass unrepaired DNA lesions. We show that TENT4A regulates mRNA stability and/or translation of DNA polymerase η and RAD18 E3 ligase, which guides the polymerase to replication stalling sites and monoubiquitinates PCNA, thereby enabling recruitment of error-prone DNA polymerases to damaged DNA sites. Remarkably, in addition to the effect on RAD18 mRNA stability via controlling its poly(A) tail, TENT4A indirectly regulates RAD18 via the tumor suppressor CYLD and via the long non-coding antisense RNA PAXIP1-AS2, which had no known function. Knocking down the expression of TENT4A or CYLD, or overexpression of PAXIP1-AS2 led each to reduced amounts of the RAD18 protein and DNA polymerase η, leading to reduced TLS, highlighting PAXIP1-AS2 as a new TLS regulator. Bioinformatics analysis revealed that TLS error-prone DNA polymerase genes and their TENT4A-related regulators are frequently mutated in endometrial cancer genomes, suggesting that TLS is dysregulated in this cancer.     

Method of Calculation of Stability Constants of Fulvic Complexes on the Example of Copper

On the example of copper fulvate, it is discussed the method of calculation of stability constants of fulvic complexes. At pH = 8, the complex formation of copper（ll） ions with fulvic acids was studied by the solubility method. Fulvic acids were separated from the river Mtkvari by the adsorption-chromotographic method. The charcoal （BAU, Russia） was used as a sorbent. The suspension of copper（If） hydroxide was used as the solid phase, on which was added the increasing quantity of standard solution of fulvic acids. In diluted solutions, at pH = 8 the dominant form of copper（lI） is copper dihydroxo complex Cu（OH）^zO. It was established that in the Cu（OH）2 （solid）-Cu（OH）^20 （solution）-FA^2-H20 system, dominates copper dyhydroxo fulvate complex with the structure 1：1, [Cu（OH）2FA]^2. The average stability constant of copper dyhydroxo fulvate complex was calculated based on experimental data fl [Cu（OH）2FAJ^2 = 4.5× l0^5.     

Investigation of the inhibitory effect of silica on the degradation of 1,1,1-trichloroethane by granular iron

Although iron-based permeable reactive barriers are gaining importance for treating groundwater contaminants, little is currently known about the effect of cosolutes on barrier longevity. Because of their corrosion inhibiting properties, dissolved silica species are of particular concern. This research investigates the effect of silica on the reduction of 1,1,1-trichloroethane by granular iron as a function of added silica concentration, pH, and duration of iron exposure to dissolved silica. Batch studies reveal that, at pH 8.5 and above, added silica concentrations as low as 0.17 mM cause a 30% reduction in the reactivity of iron. At higher silica concentrations, reactivity decreases by 65-75%. The inhibitory effect is greater at higher pH: 0.83 mM silica has no apparent adverse effect at pH 7.5, but leads to a 46% decrease in reaction rate at pH 8 and 90% at pH 9. This corresponds to observed trends in silica adsorption onto iron, which is low at pH 7.3 but increases at higher pH. Extending the duration of iron exposure to silica solutions also leads to a more pronounced inhibitory effect. This is in good agreement with the increase in silica coverage on the iron surface as revealed by X-ray photoelectron spectroscopy.     

Greedy hot-potato routing on the two-dimensional mesh

Summary We propose hot-potato (or, deflection) packet routing algorithms on the two-dimensional mesh. The algorithms are strongly greedy in the sense that they attempt to send packets in good directions whenever possible. Furthermore, the routing operations are simple and independent of the time that has elapsed. The first algorithm gives the best evacuation time known for delivering all the packets to their destinations. A batch ofk packets with maximal source-to-destination distanced _max is delivered in 2(k-1)+d _max. The second algorithm improves this bound tok+d _max when all packets are destined to the same node. This also implies a new bound for the multitarget case, which is the first to take into account the number of in-edges of a node. The third algorithm is designed for routing permutations with source-to-destination distance at most three, in which case the algorithm terminates in at most seven steps. We also show a lower bound of five steps for this problem. Ishai Ben-Aroya received the B.A. and M.Sc. in computer science from the Technion (Israel Institute of Technology). He is currently working with Microsoft Israel R&D group. His main interests include Routing Algorithms, Cryptography and Computer Security. Tamar Eilam received the B.A. degree in Computer Science from the Technion IIL in 1995, and is currently studying towards her M.A. degree. Assaf Schuster received his B.A., M.A. and Ph.D. degrees in Computer Science from the Hebrew University of Jerusalem (the last one in 1991). He is currently a lecturer at the Technion IIL. His main interests include Networks and Routing Algorithms, Parallel and Distributed Computation, Optical Computation and Communication, Dynamically Reconfiguring Networks, and Greedy Hot Potato Routing.This work was supported in part by the French-Israeli grant for cooperation in Computer Science, and by a grant from the Israeli Ministry of Science. An extended abstract appeared in proc. 2nd European Symposium on Algorithms, September 1994