Differential Expression Analysis Identifies Candidate Synaptogenic Molecules for Wiring Direction-Selective Circuits in the Retina.

An organizational feature of neural circuits is the specificity of synaptic connections. A striking example is the direction-selective (DS) circuit of the retina. There are multiple subtypes of DS retinal ganglion cells (DSGCs) that prefer motion along one of four preferred directions. This computation is mediated by selective wiring of a single inhibitory interneuron, the starburst amacrine cell (SAC), with each DSGC subtype preferentially receiving input from a subset of SAC processes. We hypothesize that the molecular basis of this wiring is mediated in part by unique expression profiles of DSGC subtypes. To test this, we first performed paired recordings from isolated mouse retinas of both sexes to determine that postnatal day 10 (P10) represents the age at which asymmetric synapses form. Second, we performed RNA sequencing and differential expression analysis on isolated P10 ON-OFF DSGCs tuned for either nasal or ventral motion and identified candidates which may promote direction-specific wiring. We then used a conditional knock-out strategy to test the role of one candidate, the secreted synaptic organizer cerebellin-4 (Cbln4), in the development of DS tuning. Using two-photon calcium imaging, we observed a small deficit in directional tuning among ventral-preferring DSGCs lacking Cbln4, though whole-cell voltage-clamp recordings did not identify a significant change in inhibitory inputs. This suggests that Cbln4 does not function primarily via a cell-autonomous mechanism to instruct wiring of DS circuits. Nevertheless, our transcriptomic analysis identified unique candidate factors for gaining insights into the molecular mechanisms that instruct wiring specificity in the DS circuit.

Analysis of structural rearrangements of poly(lactic acid) in the presence of water.

The effects of hydration on two types of structural rearrangements, physical aging and crystallization, of poly(lactic acid) (PLA) have been characterized. Although the water absorbed is extremely small, on the order of 0.5% by weight, we found a significant increase in both the magnitude and kinetics of enthalpic relaxation near the glass transition temperature (Tg). The kinetics of aging is at least 2 times the speed as compared to dry samples. In addition, we also found that the crystallization kinetics can be increased significantly when PLA is hydrated. The initial rate of crystallization at 80 °C is extremely fast at 7 J/g/min for the hydrated state as compared to 2 J/g/min under dry conditions. However, the ultimate degree of crystallinity achieved is not different for the two types of samples. Both of these structural rearrangements in the hydrated state can be explained by an increase in segmental mobility. The fact that water has such a strong effect on PLA structure is attributed to the strong intermolecular interactions present and their changes in the hydrated state. Spectroscopic features associated with bound water or free water were found for PLA with different hydration levels. The increase in the segmental mobility was directly correlated to the presence of free liquid water disrupting the intermolecular interactions in PLA.

The role of surface charge of nucleation agents on the crystallization behavior of poly(vinylidene fluoride).

The effect of the surface charge of nucleation agents on the crystallization behavior of poly(vinylidene fluoride) (PVDF) has been investigated. Ion-dipole interaction between the positive surface of nucleation agents and the partially negative CF(2) dipoles of PVDF is established as a main factor for further lowering the free energy barrier for nucleation, and thus increasing significantly the crystallization kinetics. This is in contrast to the behavior observed for nucleation agents possessing either negative surface or neutral charges. Positive nucleation agents led to a remarkable increase in the crystallization temperature of PVDF (lower supercooling) as compared with that of neat PVDF. The dispersion of each type of nucleation agent is also important. The melting temperatures of nucleation agents need to be higher than the melting temperature of PVDF. The melting point and degree of crystallinity of PVDF can also be raised by using specific nucleation agents. The detailed crystallization kinetics and conformational changes of the PVDF chain have been investigated. With the addition of positive nucleation agents, the γ and ß chain conformations, instead of the α phase, dominate.

Analysis of rapid chain dynamics in isochronal dielectric measurements of polymers.

Fast dynamics within the microwave frequency range (approximately gigahertz) in polymer systems as a function of temperature (in the range from 20 to 190 degrees C) were studied using high frequency dielectric spectroscopy. The frequency of radiation was varied from 0.5 to 18 GHz. The isochronal dielectric loss data were taken to eliminate the complexity arising from the frequency-independent, temperature-dependent background loss in the condensed phase. These studies were conducted for poly(caprolactone) (PCL), poly(ethylene oxide) (PEO), poly(ethylene oxide) with methoxy end group (PEO-CH3), PLA-b-PEO-b-PLA triblock copolymers, and several polymers with high glass transition temperatures. These polymers possess glass temperatures ranging from -62 degrees C (PCL) to 110 degrees C (PMMA). One broad relaxation process was found only for polymers (PCL, PEO, and PLA-b-PEO-b-PLA) with low glass transition temperatures. The effect due to end groups was investigated by comparing the results of PEO with hydroxy versus methoxy end groups. The measured relaxation process was determined not to be associated with end groups. The results from temperature-dependent dielectric spectroscopy indicate that the relaxation process follows an Arrhenius T dependence suggesting that it is due to local motions. The activation energy of the relaxation process was measured and investigated based on the coupling model. The results suggest that the observed relaxation process behaves as a Johari-Goldstein beta relaxation.

Versatile multilayer thin film preparation using hydrophobic interactions, crystallization, and chemical modification of poly(vinyl alcohol).

Multilayer thin film coatings were prepared on silicon substrates. Poly(vinyl alcohol) was adsorbed from aqueous solution to propyldimethylsilyl-modified silicon wafers. This thin semicrystalline coating was chemically modified using acid chlorides to form thicker, hydrophobic coatings. The products of the modification reactions allowed adsorption of a subsequent layer of poly(vinyl alcohol) that could subsequently be hydrophobized. This two-step process (adsorption/chemical modification) allows layer-by-layer deposition to prepare coatings with thickness, chemical structure, and wettability control.

Influence of copolymer configuration on the phase behavior of ternary blends.

The influence of copolymer configuration on the phase behavior of various ternary polymer blends containing a crystallizable polyester, a noncrystallizable polyether, and an acrylic random copolymer of different chain configuration was investigated. In these ternary blends, the acrylic random copolymer is typically added to control rheological properties at elevated temperatures. In fact, the acrylic random copolymers composed of various compositions of MMA and nBMA were found to have different miscibility with polyester as well as polyether, leading to substantially different phase behavior of ternary blends. Remarkable temperature dependence was also found. The mean-field Flory-Huggins theory for the free energy of mixing, extended to ternary polymer blends, was adopted for predicting phase diagrams where the exact spinodal and binodal boundaries could be calculated. Phase diagrams of ternary blends, predicted by the Flory-Huggins formulations and related calculations, were in good agreement with experimental phase diagrams. The differences observed in the rheological processes of various ternary blends with different acrylic copolymers were directly related to changes in miscibility, associated phase behavior, and chain configuration.

New bicyclic taxane diterpenoids from Taxus sumatrana.

Investigation of an acetone extract of the leaves and twigs of Taxus sumatrana has resulted in the isolation of two new bicyclic taxoids, tasumatrols M (1), and N (2) and a new baccatin III derivative, tasumatrol O (3) together with the previous known 7-deacetylcanadensene (4) and 2alpha,7beta,13alpha-triacetoxy-5alpha,9alpha-dihydroxy-2(3-->20) abeotaxa-4(20),11-dien-10-one. The structures of these taxanes were established on the basis of spectroscopic analyses, especially 1- and 2D NMR, and chemical derivatization.

Vibrational spectrum, ab initio calculations, conformational equilibria and torsional modes of 1,3-dichloropropane.

Ab initio calculations are reported for three of four possible conformers of 1,3-dichloropropane. The fourth conformer, with Cs symmetry, has a predicted enthalpy difference of more than 1500 cm(-1) from the most stable conformer from each calculation regardless of the basis set used, so there is little chance of observing it. Thus, there is no evidence in the infrared or Raman spectrum of the presence of a fourth conformer. The order of stability given by the ab initio calculations is C2(GG)>C1(AG)>C2v(AA)>Cs(GG'), where A indicates the anti form for one of the CH2Cl groups and G indicates the gauche conformation for the other CH2Cl group relative to the plane of the carbon atoms. Almost every band observed can be confidently assigned to one or another of the conformers. Many observed bands proved to be of a composite nature, with several nearly coincident vibrations of different conformers contributing to the band contour. Nonetheless, a complete assignment of fundamentals is possible for the most stable C2 conformer, and 5 of the fundamentals of the C2v conformer and 13 those of the C1 conformer can be confidently assigned.

Binding efficiency and transport properties of molecularly imprinted polymer thin films.

A model system for the characterization of molecular recognition events in molecularly imprinted polymers (MIPs) is presented. The use of a biologically inspired, three-point hydrogen-bonding motif and a thin film polymeric matrix allows for pre- and post-polymerization binding properties to be characterized by infrared spectroscopy. A method to determine binding constants was developed and utilized before and after cross-linking. These values showed a 10-fold decrease in binding after polymerization, which was attributed to an increase in molecular confinement after polymerization and a change in the local structural environment of the binding cavity. Transport of the guest molecule was shown to be reversible.