Super-Resolution STED Microscopy-Based Mobility Studies of the Viral Env Protein at HIV-1 Assembly Sites of Fully Infected T-Cells

The ongoing threat of human immunodeficiency virus (HIV-1) requires continued, detailed investigations of its replication cycle, especially when combined with the most physiologically relevant, fully infectious model systems. Here, we demonstrate the application of the combination of stimulated emission depletion (STED) super-resolution microscopy with beam-scanning fluorescence correlation spectroscopy (sSTED-FCS) as a powerful tool for the interrogation of the molecular dynamics of HIV-1 virus assembly on the cell plasma membrane in the context of a fully infectious virus. In this process, HIV-1 envelope glycoprotein (Env) becomes incorporated into the assembling virus by interacting with the nascent Gag structural protein lattice. Molecular dynamics measurements at these distinct cell surface sites require a guiding strategy, for which we have used a two-colour implementation of sSTED-FCS to simultaneously target individual HIV-1 assembly sites via the aggregated Gag signal. We then compare the molecular mobility of Env proteins at the inside and outside of the virus assembly area. Env mobility was shown to be highly reduced at the assembly sites, highlighting the distinct trapping of Env as well as the usefulness of our methodological approach to study the molecular mobility of specifically targeted sites at the plasma membrane, even under high-biosafety conditions.     

Amorphous Silicon Oxynitride-Based Powders Produced by Spray Pyrolysis from Liquid Organosilicon Compounds

Silicon oxynitrides (SiO_xN_y) have many advantageous properties for modern ceramic applications that justify a development of their new and efficient preparation methods. In the paper, we show the possibility of preparing amorphous SiO_xN_y-based materials from selected liquid organosilicon compounds, methyltrimethoxysilane CH₃Si(OCH₃)₃ and methyltriethoxysilane CH₃Si(OC₂H₅)₃, by a convenient spray pyrolysis method. The precursor mist is transported with an inert gas or a mixture of reactive gases through a preheated tube reactor to undergo complex decomposition changes, and the resulting powders are collected in the exhaust filter. The powders are produced in the tube at temperatures of 1200, 1400, and 1600 °C under various gas atmosphere conditions. In the first option, argon Ar gas is used for mist transportation and ammonia NH₃ gas serves as a reactive medium, while in the second option nitrogen N₂ is exclusively applied. Powder X-Ray Diffraction (XRD) results confirm the highly amorphous nature of all products except those made at 1600 °C in nitrogen. SEM examination shows the spheroidal particle morphology of powders, which is typical for this method. Fourier Transform Infrared (FT-IR) spectroscopy reveals the presence of Si–N and Si–O bonds in the powders prepared under Ar/NH₃, whereas those produced under N₂ additionally contain Si–C bonds. Raman spectroscopy measurements also support some turbostratic free carbon C in the products prepared under nitrogen. The directly determined O- and N-contents provide additional data linking the process conditions with specific powder composition, especially from the point of view of oxygen replacement in the Si–O moieties formed upon initial precursor decomposition reactions by nitrogen (from NH₃ or N₂) or carbon (from the carbonization of the organic groups).     

Wear Resistance Improvement of Cemented Tungsten Carbide Deep-Hole Drills after Ion Implantation

The paper is dedicated to the life prolongation of the tools designed for deep-hole drilling. Among available methods, an ion implantation process was used to improve the durability of tungsten carbide (WC)-Co guide pads. Nitrogen fluencies of 3 × 10¹⁷ cm⁻², 4 × 10¹⁷ cm⁻² and 5 × 10¹⁷ cm⁻² were applied, and scanning electron microscope (SEM) observations, energy dispersive spectroscopy (EDS) analyses, X-ray photoelectron spectroscopy (XPS) and Secondary Ion Mass Spectrometry (SIMS) measurements were performed for both nonimplanted and implanted tools. The durability tests of nonimplanted and the modified tools were performed in industrial conditions. The durability of implanted guide pads was above 2.5 times greater than nonimplanted ones in the best case, presumably due to the presence of a carbon-rich layer and extremely hard tungsten nitrides. The achieved effect may be attributed to the dissociation of tungsten carbide phase and to the lubrication effect. The latter was due to the presence of pure carbon layer with a thickness of a few dozen nanometers. Notably, this layer was formed at a temperature of 200 °C, much smaller than in previously reported research, which makes the findings even more valuable from economic and environmental perspectives.     

The aromatase inhibitor anastrozole is ineffective in the treatment of precocious puberty in girls with McCune-Albright syndrome

CONTEXT: Precocious puberty (PP) in girls with McCune-Albright syndrome (MAS) is characterized by episodic development of large unilateral ovarian cysts followed by sudden onset of vaginal bleeding. Some patients experience frequent bleeding as well as accelerated linear growth and advanced skeletal maturation. The use of anastrozole for the treatment of PP in this condition has not been well studied. OBJECTIVE: The objective of the study was to determine the safety and efficacy of the aromatase inhibitor anastrozole for the treatment of PP in girls with MAS. DESIGN AND SETTINGS: This was a prospective international multicenter study in which subjects received anastrozole 1 mg daily for 1 yr. PATIENTS: Twenty-eight girls 10 years of age or younger with MAS and progressive PP were enrolled. MAIN OUTCOME MEASURES: Vaginal bleeding, rate of skeletal maturation (change in bone age over change in chronological age), growth velocity, and uterine/ovarian volumes were measured. These indices were compared with a 6-month pretreatment interval. RESULTS: No difference in vaginal bleeding (mean number of days per year) was noted. Mean change in DeltaBA/DeltaCA, which was 1.25 +/- 0.77 at baseline, was -0.25 +/- 1.02 at study end (P = 0.22). Average growth velocity z score was 1.40 +/- 3.15 at study entry and 0.26 +/- 2.71 at 12 months (P = 0.10). Mean ovarian/uterine volumes were unaffected by anastrozole, and no significant adverse events occurred. CONCLUSIONS: Although it appears safe, anastrozole for 1 yr was ineffective in halting vaginal bleeding, attenuating rates of skeletal maturation, and linear growth in girls with MAS. Pharmacological strategies other than anastrozole should be pursued for the treatment of PP in this population.     

Optimization and regeneration kinetics of lymphatic-specific photodynamic therapy in the mouse dermis

Lymphatic vessels transport fluid, antigens, and immune cells to the lymph nodes to orchestrate adaptive immunity and maintain peripheral tolerance. Lymphangiogenesis has been associated with inflammation, cancer metastasis, autoimmunity, tolerance and transplant rejection, and thus, targeted lymphatic ablation is a potential therapeutic strategy for treating or preventing such events. Here we define conditions that lead to specific and local closure of the lymphatic vasculature using photodynamic therapy (PDT). Lymphatic-specific PDT was performed by irradiation of the photosensitizer verteporfin that effectively accumulates within collecting lymphatic vessels after local intradermal injection. We found that anti-lymphatic PDT induced necrosis of endothelial cells and pericytes, which preceded the functional occlusion of lymphatic collectors. This was specific to lymphatic vessels at low verteporfin dose, while higher doses also affected local blood vessels. In contrast, light dose (fluence) did not affect blood vessel perfusion, but did affect regeneration time of occluded lymphatic vessels. Lymphatic vessels eventually regenerated by recanalization of blocked collectors, with a characteristic hyperplasia of peri-lymphatic smooth muscle cells. The restoration of lymphatic function occurred with minimal remodeling of non-lymphatic tissue. Thus, anti-lymphatic PDT allows control of lymphatic ablation and regeneration by alteration of light fluence and photosensitizer dose.     

A graphene-based hybrid material with quantum bits prepared by the double Langmuir–Schaefer method

The scalability and stability of molecular qubits deposited on surfaces is a crucial step for incorporating them into upcoming electronic devices. Herein, we report on the preparation and characterisation of a molecular quantum bit, copper(ii)dibenzoylmethane [Cu(dbm)₂], deposited by a modified Langmuir–Schaefer (LS) technique onto a graphene-based substrate. A double LS deposition was used for the preparation of a few-layer-graphene (FLG) on a Si/SiO₂ substrate with subsequent deposition of the molecules. Magnetic properties were probed by high-frequency electron spin resonance (HF-ESR) spectroscopy and found maintained after deposition. Additional spectroscopic and imaging techniques, such as Raman spectroscopy (RS), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and scanning electron microscopy (SEM) were performed to characterise the deposited sample. Our approach demonstrated the possibility to utilise a controlled wet-chemistry protocol to prepare an array of potential quantum bits on a disordered graphene-based substrate. The deployed spectroscopic techniques showed unambiguously the robustness of our studied system with a potential to fabricate large-scale, intact, and stable quantum bits.

Graphene-based hybrid material with array of copper(ii)-based quantum bits was prepared by a wet-chemistry protocol and characterised by HF-ESR, XPS, Raman, and AFM.     

Synthesis,structural studies and biological properties of some phosphono-perfluorophenylalanine derivatives formed by SNAr reactions

Several novel phosphono-perfluorophenylalanine derivatives, as mimetics of phenylalanine, were synthesized by subjecting diethyl (2-(perfluorophenyl)-1-(phenylamino)ethyl)-phosphonate to S_NAr reactions with different types of nucleophiles such as thiols, amines and phenols. The structure of the products was confirmed using spectroscopic and spectrometric techniques. For two compounds X-ray single crystal diffraction analysis and DFT investigations were performed providing information in regard to the preferable conformation, hydrogen bonds and other interactions. The antiproliferative potency of some of the new phosphono-perfluorophenylalanine derivatives obtained as well as representatives of previously synthesized perfluorophenyl phosphonate analogues of phenylalanine was studied on selected glioma cell lines. Preliminary evaluation of the compounds drug likeness was examined with respect to Lipinski''s and Veber''s rules, and showed that they meet the criteria perfectly. MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay results demonstrated that the compounds exhibit moderate activity against the glioblastoma multiforme cell lines (T98G and U-118 MG). Moreover most of the studied S_NAr reaction products displayed significantly higher inhibitory activity against both cancer cell lines than the parent diethyl (2-(perfluorophenyl)-1-(phenylamino)ethyl)phosphonate.

Several novel phosphono-perfluorophenylalanine derivatives, as mimetics of phenylalanine, were synthesized by subjecting diethyl (2-(perfluorophenyl)-1-(phenylamino)ethyl)-phosphonate to S_NAr reactions with thiols, amines and phenols.