Customizable Lipid Nanoparticle Materials for the Delivery of siRNAs and mRNAs

RNAs are a promising class of therapeutics given their ability to regulate protein concentrations at the cellular level. Developing safe and effective strategies to deliver RNAs remains important for realizing their full clinical potential. Here, we develop lipid nanoparticle formulations that can deliver short interfering RNAs (for gene silencing) or messenger RNAs (for gene upregulation). Specifically, we study how the tail length, tail geometry, and linker spacing in diketopiperazine lipid materials influences LNP potency with siRNAs and mRNAs. Eight lipid materials are synthesized, and 16 total formulations are screened for activity in vitro; the lead material is evaluated with mRNA for in vivo use and demonstrates luciferase protein expression in the spleen. In undertaking this approach, not only do we develop synthetic routes to delivery materials, but we also reveal structural criteria that could be useful for developing next‐generation delivery materials for RNA therapeutics.     

A Criterion for Finite Topological Determinacy of Map-Germs

Let f, g: (Rⁿ, 0) (R^p, 0) be two C map-germs. Then f and gare C⁰-equivalent if there exist homeomorphism-germs h and lof (Rⁿ, 0) and (R^p, 0) respectively such that g = l f h^–1.Let k be a positive integer. A germ f is k-C⁰-determined ifevery germ g with j^k g(0) = j^k f(0) is C⁰-equivalent to f. Moreover,we say that f is finitely topologically determined if f is k-C⁰-determinedfor some finite k. We prove a theorem giving a sufficient conditionfor a germ to be finitely topologically determined. We explainthis condition below. Let N and P be two C manifolds. Consider the jet bundle J^k(N,P) with fiber J^k(n, p). Let z in J^k(n, p) and let f be suchthat z = j^kf(0). Define Whether (f) < k depends only on z, not on f. We can thereforedefine the set Let W^k(N, P) be the subbundle of J^k(N, P) with fiber W^k(n, p).Mather has constructed a finite Whitney (b)-regular stratificationS^k(n, p) of J^k(n, p) – W^k(n, p) such that all strata aresemialgebraic and K-invariant, having the property that if S^k(N,P) denotes the corresponding stratification of J^k(N, P) –W^k(N, P) and f C(N, P) is a C map such that j^kf is multitransverseto S^k(N, P), j^kf(N) W^k(N, P) = and N is compact (or f is proper),then f is topologically stable. For a map-germ f: (Rⁿ, 0) (R^p, 0), we define a certain ojasiewiczinequality. The inequality implies that there exists a representativef: U R^p such that j^kf(U – 0) W^k (Rⁿ, R^p = and suchthat j^kf is multitransverse to S^k (Rⁿ, R^p) at any finite setof points S U – 0. Moreover, the inequality controlsthe rate j^kf becomes non-transverse as we approach 0. We showthat if f satisfies this inequality, then f is finitely topologicallydetermined. 1991 Mathematics Subject Classification: 58C27.     

Micropatterning of functional conductive polymers with multiple surface chemistries in register

A versatile procedure is presented for fast and efficient micropatterning of multiple types of covalently bound surface chemistry in perfect register on and between conductive polymer microcircuits. The micropatterning principle is applied to several types of native and functionalized PEDOT (poly(3,4-ethylenedioxythiophene)) thin films. The method is based on contacting PEDOT-type thin films with a micropatterned agarose stamp containing an oxidant (aqueous hypochlorite) and applying a nonionic detergent. Where contacted, PEDOT not only loses its conductance but is entirely removed, thereby locally revealing the underlying substrate. Surface analysis showed that the substrate surface chemistry was fully exposed and not affected by the treatment. Click chemistry could thus be applied to selectively modify re-exposed alkyne and azide functional groups of functionalized polystyrene substrates. The versatility of the method is illustrated by micropatterning cell-binding RGD-functionalized PEDOT on low cell-binding PMOXA (poly(2-methyl-2-oxazoline)) to produce cell-capturing microelectrodes on a cell nonadhesive background in a few simple steps. The method should be applicable to a wide range of native and chemically functionalized conjugated polymer systems.     

Fluorescent Saxitoxins for Live Cell Imaging of Single Voltage-Gated Sodium Ion Channels beyond the Optical Diffraction Limit

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Highlights? Fluorescent saxitoxins synthesized as imaging agents for sodium ion channels (Na_Vs) ? Electrophysiology and confocal imaging establish potency and selectivity of Na_V probes ? Single-molecule and super-resolution imaging reveal subdiffraction Na_V distributions ? Dynamic motions of Na_Vs measured in specific membrane regions in live cells     

Internal Rotation of the Acetyl Methyl Group in Methyl Alkyl Ketones: The Microwave Spectrum of Octan-2-one

Methyl n-alkyl ketones form a class of molecules with interesting internal dynamics in the gas-phase. They contain two methyl groups undergoing internal rotations, the acetyl methyl group and the methyl group at the end of the alkyl chain. The torsional barrier of the acetyl methyl group is of special importance, since it allows for the discrimination of the conformational structures. As part of the series, the microwave spectrum of octan-2-one was recorded in the frequency range from 2 to 40 GHz, revealing two conformers, one with C₁ and one with C_s symmetry. The barriers to internal rotation of the acetyl methyl group were determined to be 233.340(28) cm⁻¹ and 185.3490(81) cm⁻¹, respectively, confirming the link between conformations and barrier heights already established for other methyl alkyl ketones. Extensive comparisons to molecules in the literature were carried out, and a small overview of general trends and rules concerning the acetyl methyl torsion is given. For the hexyl methyl group, the barrier height is 973.17(60) cm⁻¹ for the C₁ conformer and 979.62(69) cm⁻¹ for the C_s conformer.     

The phase shift method applied to cross grating moiré measurements

The automatic evaluation of the grating deformation in a moiré image is greatly enhanced by the phase shift method. Based on three or more local grey values of a related number of phase shifted moiré patterns, the local displacement of a deformed grating against a reference grating, and hence the total displacement in one direction, is calculated. In this paper an extension to cross gratings is presented yielding a two- dimensional displacement field.     

Current trends in finite-time thermodynamics

The cornerstone of finite-time thermodynamics is all about the price of haste and how to minimize it. Reversible processes may be ultimately efficient, but they are unrealistically slow. In all situations-chemical, mechanical, economical-we pay extra to get the job done quickly. Finite-time thermodynamics can be used to develop methods to limit that extra expenditure, be it in energy, entropy production, money, or something entirely different. Finite-time thermodynamics also includes methods to calculate the optimal path or mode of operation to achieve this minimal expenditure. The concept is to place the system of interest in contact with a time-varying environment which will coax the system along the desired path, much like guiding a horse along by waving a carrot in front of it.     

Stimulated Raman scattering microscopy by spectral focusing and fiber-generated soliton as Stokes pulse

We demonstrate stimulated Raman microscopy with broadband pump and Stokes pulses, using spectral focusing to attain spectral resolution and to rapidly acquire spectra within a spectral window determined by the bandwidth of the pulses. As the Stokes pulse, we use the redshifted soliton generated in a photonic crystal fiber, which allows for simple shifting of the accessible spectral window.