Assessing Photostability of mAb Formulations In Situ Using Light-Coupled NMR Spectroscopy.

Biopharmaceuticals, such as monoclonal antibodies (mAbs), need to maintain their chemical and physical stability in formulations throughout their lifecycle. It is known that exposure of mAbs to light, particularly UV, triggers chemical and physical degradation, which can be exacerbated by trace amounts of photosensitizers in the formulation. Although routine assessments of degradation following defined UV dosages are performed, there is a fundamental lack of understanding regarding the intermediates, transient reactive species, and radicals formed during illumination, as well as their lifetimes and immediate impact post-illumination.

Photochemistry of Ru(II) Triazole Complexes with 6-Membered Chelate Ligands: Detection and Reactivity of Ligand-Loss Intermediates.

Photochemical ligand release from metal complexes may be exploited in the development of novel photoactivated chemotherapy agents for the treatment of cancer and other diseases. Highly intriguing photochemical behavior is reported for two ruthenium(II) complexes bearing conformationally flexible 1,2,3-triazole-based ligands incorporating a methylene spacer to form 6-membered chelate rings. [Ru(bpy)(pictz)] () and [Ru(bpy)(btzm)] () (bpy = 2,2'-bipyridyl; pictz = 1-(picolyl)-4-phenyl-1,2,3-triazole; btzm = bis(4-phenyl-1,2,3-triazol-4-yl)methane) exhibit coordination by the triazole ring through the less basic N2 atom as a consequence of chelation and readily undergo photochemical release of the pictz and btzm ligands (ϕ = 0.

Use of spatiotemporal couplings and an axiparabola to control the velocity of peak intensity.

This paper presents the first experimental realization of a scheme that allows for the tuning of the velocity of peak intensity of a focal spot with relativistic intensity. By combining a tunable pulse-front curvature with the axial intensity deposition characteristics of an axiparabola, an aspheric optical element, this system provides control over the dynamics of laser-wakefield accelerators. We demonstrate the ability to modify the velocity of peak intensity of ultrashort laser pulses to be superluminal or subluminal.

Simple and effective sample illumination for electron paramagnetic resonance.

Illumination into an electron paramagnetic resonance (EPR) spectrometer is commonly carried out through the optical window, perpendicular to the sample and magnetic field. Here we show that significant improvements can be obtained by using the walls of the EPR tube as a light guide, with the light scattered only around the sample-containing area.

Different Structures-Similar Effect: Do Substituted 5-(4-Methoxyphenyl)-1-indoles and 5-(4-Methoxyphenyl)-1-imidazoles Represent a Common Pharmacophore for Substrate Selective Inhibition of Linoleate Oxygenase Activity of ALOX15?

Mammalian 15-lipoxygenases (ALOX15) are lipid peroxidizing enzymes that exhibit variable functionality in different cancer and inflammation models. The pathophysiological role of linoleic acid- and arachidonic acid-derived ALOX15 metabolites rendered this enzyme a target for pharmacological research. Several indole and imidazole derivatives inhibit the catalytic activity of rabbit ALOX15 in a substrate-specific manner, but the molecular basis for this allosteric inhibition remains unclear.

Microwave Diamond-Based HBAR as a Highly Sensitive Sensor for Multiple Applications: Acoustic Attenuation in the Mo Film.

The application of microwave diamond-based HBAR as a sensor of microwave acoustic attenuation α was considered, using the Mo film as an object of research. A multilayered piezoelectric structure, as the Al/AlScN/Mo/(100) diamond/Mo, was produced using aluminum-scandium nitride composition, and was studied in detail for a number of the Mo films with different thicknesses obtained by magnetron deposition. The operational frequency band of 3.

Chemometrics in Protein Formulation: Stability Governed by Repulsion and Protein Unfolding.

Therapeutic proteins can be challenging to develop due to their complexity and the requirement of an acceptable formulation to ensure patient safety and efficacy. To date, there is no universal formulation development strategy that can identify optimal formulation conditions for all types of proteins in a fast and reliable manner. In this work, high-throughput characterization, employing a toolbox of five techniques, was performed on 14 structurally different proteins formulated in 6 different buffer conditions and in the presence of 4 different excipients.

Energy-Conserving Theory of the Blowout Regime of Plasma Wakefield.

We present a self-consistent theory of strongly nonlinear plasma wakefield (bubble or blowout regime of the wakefield) based on the energy conservation approach. Such wakefields are excited in plasmas by intense laser or particle beam drivers and are characterized by the expulsion of plasma electrons from the propagation axis of the driver. As a result, a spherical cavity devoid of electrons (called a "bubble") and surrounded by a thin sheath made of expelled electrons is formed behind the driver.

Proteolysis of Micellar β-Casein by Trypsin: Secondary Structure Characterization and Kinetic Modeling at Different Enzyme Concentrations.

Tryptic proteolysis of protein micelles was studied using β-casein (β-CN) as an example. Hydrolysis of specific peptide bonds in β-CN leads to the degradation and rearrangement of the original micelles and the formation of new nanoparticles from their fragments. Samples of these nanoparticles dried on a mica surface were characterized by atomic force microscopy (AFM) when the proteolytic reaction had been stopped by tryptic inhibitor or by heating.

Cyclization of arylhydrazones of cross-conjugated enynones: synthesis of luminescent styryl-1-pyrazoles and propenyl-1-pyrazoles.

Condensation of 1,5-disubstituted pent-1-en-4-yn-1-ones with arylhydrazines in acidified alcohol results mainly in the formation of the corresponding arylhydrazones with traces of the side products of cyclization at the double bond - 1,5-diaryl-3-(arylethynyl)-4,5-dihydro-1-pyrazoles (pyrazolines). Arylhydrazones are cyclized only by refluxing in high-boiling polar solvents (DMF and ethylene glycol), with the selective formation of 1,5-disubstituted 3-styrylpyrazoles in up to 77-95% yields. Thermodynamically, the cyclization of arylhydrazones at the triple bond is the most preferable pathway, as shown by DFT calculations and preparative synthesis experiments.

Lipopolysaccharide Structure and the Phenomenon of Low Endotoxin Recovery.

Lipopolysaccharide (LPS) is a cell-wall component of Gram-negative bacteria which contributes to bacterial toxicity. During processes such as cell division, shedding of outer membrane vesicles, or bacterial cell death, LPS is released into the surrounding media. If such contamination got into the bloodstream, it would induce pro-inflammatory immune responses which can result in sepsis and death.

Insights into the Stabilization of Interferon Alpha by Two Surfactants Revealed by STD-NMR Spectroscopy.

Surfactants are commonly used in biopharmaceutical formulations to stabilize proteins against aggregation. However, the choice of a suitable surfactant for a particular protein is decided mostly empirically, and their mechanism of action on molecular level is largely unknown. Here we show that a straightforward label-free method, saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy, can be used to detect protein-surfactant interactions in formulations of a model protein, interferon alpha.

Controlled release and characterisation of photocaged molecules using LED illumination in solution NMR spectroscopy.

Photocaging is an attractive strategy to control molecular behaviour, for example, in chemical synthesis, interaction studies or photodynamic therapies. Here, we demonstrate that illumination by the LED NMRtorch approach enables effective and controlled photocage release with simultaneous monitoring of subsequent reactions by solution NMR spectroscopy.

Positron acceleration via laser-augmented blowouts in two-column plasma structures.

We propose a setup for positron acceleration consisting of an electron driver and a laser pulse creating a twofold plasma column structure. The resulting wakefield is capable of accelerating positron bunches over long distances even when the evolution of the driver is considered. The scheme is studied by means of particle-in-cell simulations.

Ketone Derivatives of Propargylamines as Synthetic Equivalents of Conjugated 2,4,1-Enynones in the Synthesis of Acetylenic 2-Pyrazolines and Pyrazoles.

An interaction of 1,5-diaryl-3-X-pent-4-yn-1-ones (where X stands for piperidin-1-yl, morpholin-4-yl, 4-methylpiperazin-1-yl) with arylhydrazines proceeds at room temperature and results in 3-aryl-5-arylethynyl-1-phenyl-4,5-dihydro-1-pyrazoles with up to 57-73% yields. Under similar conditions, the cyclocondensation of conjugated 2,4,1-enynones with arylhydrazine proceeds only in the presence of cyclic amines. 1,5-Diaryl-3-X-pent-4-yn-1-ones are reported as synthetic equivalents of conjugated 2,4,1-enynones in reactions with arylhydrazines.

Temporal and spatial characterisation of protein liquid-liquid phase separation using NMR spectroscopy.

Liquid-liquid phase separation (LLPS) of protein solutions is increasingly recognised as an important phenomenon in cell biology and biotechnology. However, opalescence and concentration fluctuations render LLPS difficult to study, particularly when characterising the kinetics of the phase transition and layer separation. Here, we demonstrate the use of a probe molecule trifluoroethanol (TFE) to characterise the kinetics of protein LLPS by NMR spectroscopy.

[Lesions of the cortical part of the auditory analyzer in explosive injury].

A survey of 48 victims aged 19-36 years with explosive trauma and combined damage to the auditory system was conducted to assess the level of damage to nerve structures by analyzing the bioelectric activity of the cerebral cortex. All patients underwent electroencephalography (EEG). It is established that akubarotrauma of explosive genesis almost always leads to lesions of the function of the cortical part of the auditory analyzer.

Michael Addition of 3-Oxo-3-phenylpropanenitrile to Linear Conjugated Enynones: Approach to Polyfunctional δ-Diketones as Precursors for Heterocycle Synthesis.

Reaction of linear conjugated enynones, 1,5-diarylpent-2-en-4-yn-1-ones [ArC≡CCH=CHC(=O)Ar], with 3-oxo-3-phenylpropanenitrile (NCCHCOPh) in the presence of sodium methoxide MeONa as a base in MeOH at room temperature for 4-26 h affords polyfunctional δ-diketones as a product of regioselective Michael addition to the double carbon-carbon bond of starting enynones. The δ-diketones have been formed as mixtures of two diastereomers in a ratio of 2.5:1 in good general yields of 53-98%.

-Substituted 5-(1-Indol-2-yl)-2-methoxyanilines Are Allosteric Inhibitors of the Linoleate Oxygenase Activity of Selected Mammalian ALOX15 Orthologs: Mechanism of Action.

Here, we describe the first systematic study on the mechanism of substrate-selective inhibition of mammalian ALOX15 orthologs. For this purpose, we prepared a series of -substituted 5-(1-indol-2-yl)anilines and found that (-(5-(1-indol-2-yl)-2-methoxyphenyl)sulfamoyl)carbamates and their monofluorinated analogues are potent and selective inhibitors of the linoleate oxygenase activity of rabbit and human ALOX15. Introduction of a 2-methoxyaniline moiety into the core pharmacophore plays a crucial role in substrate-selective inhibition of ALOX15-catalyzed oxygenation of linoleic acid at submicromolar concentrations without affecting arachidonic acid oxygenation.

Efficient Narrow-Band Terahertz Radiation from Electrostatic Wakefields in Nonuniform Plasmas.

It is shown that electrostatic plasma wakefields can efficiently radiate at harmonics of the plasma frequency when the plasma has a positive density gradient along the propagation direction of a driver. The driver propagating at a subluminal group velocity excites the plasma wakefield with the same phase velocity. However, due to the positive density gradient, the wake phase velocity steadily increases behind the driver.

Stability of a high-concentration monoclonal antibody solution produced by liquid-liquid phase separation.

Subcutaneous injection of a low volume (<2 mL) high concentration (>100 mg/mL) formulation is an attractive administration strategy for monoclonal antibodies (mAbs) and other biopharmaceutical proteins. Using concentrated solutions may also be beneficial at various stages of bioprocessing. However, concentrating proteins by conventional techniques, such as ultrafiltration, can be time consuming and challenging.

Functionalized Homologues and Positional Isomers of Rabbit 15- Lipoxygenase RS75091 Inhibitor.

Background: RS75091 is a cinnamic acid derivative that has been used for the crystallization of the rabbit ALOX15-inhibitor complex. The atomic coordinates of the resolved ALOX15- inhibitor complex were later on used to define the binding sites of other mammalian lipoxygenase orthologs, for which no direct structural data with ligand has been reported so far.

Introduction: The putative binding pocket of the human ALOX5 was reconstructed on the basis of its structural alignment with rabbit ALOX15-RS75091 inhibitor.