A Novel Elvitegravir Nanoformulation for Drug Delivery Across the Blood-Brain Barrier to Suppress HIV-1 in Macrophages and Microglia

Over the last two decades, the use of antiretroviral therapy (ART) has remarkably decreased the morbidity associated with HIV-1 infection. However, the prevalence of HIV-1-associated neurocognitive disorders (HAND) is still increasing. The appearance and persistence of HAND are partially due to the entry of HIV-1-infected monocytes into the brain. Within the central nervous system (CNS), HIV-1-infected macrophages and microglia serve as the major viral reservoirs and provide active viral replication even when systemic viral suppression has been achieved by ART. Efficient viral suppression of CNS macrophages and microglia is important for an effective HIV-1 treatment in the brain. The primary hindrance to the treatment of HAND is mainly contributed by the inability of antiretrovirals (ARVs) to cross the blood-brain barrier (BBB) after the systemic administration. Conventional regimens of ARVs are not sufficient to penetrate the BBB or improve outcomes in HAND. Thus, there is a need for novel treatment regimens that cross the BBB and deliver therapeutic ARVs into the CNS to suppress the HIV-1 replication in these viral reservoirs. Our objective in this study was to improve the efficacy of ART in CNS HIV-1 reservoirs, specifically in macrophages and microglia. Our central hypothesis was that the selected ARV, elvitegravir (EVG), in nanoformulation would have increased transmigration across the BBB, as well as increased drug uptake in macrophages and microglia after crossing the BBB, leading to increased antiviral activity in macrophages and microglia, relative to the native drug. This study was expected to provide an optimized treatment strategy that has potential for therapeutic interventions in reducing HAND.In this study, poly (lactic-co-glycolic acid) (PLGA)-based elvitegravir nanoparticles (PLGA-EVG NPs) were prepared by nano-precipitation. Firstly, we analyzed the physicochemical properties of PLGA-EVG NPs using transmission electron microscopy, dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FTIR). We measured cellular uptake of PLGA NPs by fluorescence microscopy and flow cytometry. We also measured the intracellular drug concentration and viral replication in HIV-1-infected macrophages by using LC-MS/MS and p24 ELISA, respectively. The PLGA-

Plasmonic nano-resonator enhanced one-photon luminescence from single gold nanorods

Strong Stokes and anti-Stokes one-photon luminescence from single gold nanorods is measured in experiments. It is found that the intensity and polarization of the Stokes and anti-Stokes emissions are in strong correlation. Our experimental observation discovered a coherent process in light emission from single gold nanorods. We present a theoretical mode, based on the concept of cavity resonance, for consistently understanding both Stokes and anti-Stokes photoluminescence. Our theory is in good agreement of all our measurements.Comment: 14 pages, 7 figures, 2 table

Highly sensitive magnetic properties and large linear magnetoresistance in antiferromagnetic CrxSe(0.875\lex\le1)single crystals

CrxSe (x\le1) is a class of quasi-layered binary compounds with potential applications in spintronics due to its intriguing antiferromagnetic properties. In this work, CrxSe single crystals with high Cr content (x=0.87, 0.91 and 0.95) were grown, and their magnetic and transport properties were investigated in detail. It is found that with small increase of Cr content, the N\'eel temperature (TN) of the samples can dramatically increase from 147 K to 257 K, accompanied with obvious changes in the magnetic anisotropy and hysteresis. The phenomena of field-induced spin-flop transitions were unveiled in these alloys, indicating their comparatively low anisotropy. The magnetoresistance (MR) of the three compounds showed positive dependence at low temperatures and particularly, non-saturated linear positive MR was observed in Cr0.91Se and Cr0.95Se, with a large value of 16.2% achieved in Cr0.91Se (10K, 9T). The calculated Fermi surface and MR showed that the quasi-linear MR is a product of carrier compensation. Our work revealed highly sensitive magnetic and transport properties in the Cr-Se compounds, which can lay foundation when constructing further antiferromagnetic spintronic devices based on them

A Framework for Calculating the Failure Probability of Natural Gas Pipeline

Reliability based design and assessment (RBDA) technique is a developing direction of natural gas pipeline design method. In this paper, a framework for calculating the failure probability of natural gas pipeline is proposed. First, Java reflection mechanism is used in the management of the limit state functions, which enables the separation of the limit state algorithms and the calculations of the failure probability. Under this framework, more newly developed equations can be add into the library of the software readily. Second, a Monte Carlo reliability analysis algorithm capable of incorporating the basic input parameters and limit state functions is used to calculate failure probability of pipelines. Third, a post data processing algorithm is used to improve the efficiency. Finally, an example on natural gas pipeline is presented to illustrate the availability and effectiveness of the software. Experimental results indicate the ability of the proposed framework for pipeline quality control

Directional Enhanced Probe for Side-Illumination Tip Enhanced Spectroscopy

We demonstrate a high-performance apertureless near-field probe made of a tapered metal tip with a set of periodic shallow grooves near the apex. The spontaneous emission from a single emitter near the tip is investigated systematically for the side-illumination tip enhanced spectroscopy (TES). In contrast with the bare tapered metal tip in conventional side-illumination TES, the corrugated probe not only enhances strongly local excitation field but also concentrates the emission directivity, which leads to high collection efficiency and signal-to-noise ratio. In particular, we propose an asymmetric TES tip based on two coupling nanorods with different length at the apex to realize unidirectional enhanced emission rate from a single emitter. Interestingly, we find that the radiation pattern is sensitive to the emission wavelength and the emitter positions respective to the apex, which can result in an increase of signal-to-noise ratio by suppressing undesired signal. The proposed asymmetrical corrugated probe opens up a broad range of practical applications, e.g. increasing the detection efficiency of tip enhanced spectroscopy at the single-molecule level

Novel elvitegravir nanoformulation for drug delivery across the blood-brain barrier to achieve HIV-1 suppression in the CNS macrophages

The use of antiretroviral therapy (ART) has remarkably decreased the morbidity associated with HIV-1 infection, however, the prevalence of HIV-1-associated neurocognitive disorders (HAND) is still increasing. The blood-brain barrier (BBB) is the major impediment for penetration of antiretroviral drugs, causing therapeutics to reach only suboptimal level to the brain. Conventional antiretroviral drug regimens are not sufficient to improve the treatment outcomes of HAND. In our recent report, we have developed a poloxamer-PLGA nanoformulation loaded with elvitegravir (EVG), a commonly used antiretroviral drug. The nanoformulated EVG is capable of elevating intracellular drug uptake and simultaneously enhance viral suppression in HIV-1-infected macrophages. In this work, we identified the clinical parameters including stability, biocompatibility, protein corona, cellular internalization pathway of EVG nanoformulation for its potential clinical translation. We further assessed the ability of this EVG nanoformulation to cross the in vitro BBB model and suppress the HIV-1 in macrophage cells. Compared with EVG native drug, our EVG nanoformulation demonstrated an improved BBB model penetration cross the in vitro BBB model and an enhanced HIV-1 suppression in HIV-1-infected human monocyte-derived macrophages after crossing the BBB model without altering the BBB model integrity. Overall, this is an innovative and optimized treatment strategy that has a potential for therapeutic interventions in reducing HAND

Salt tolerance analysis of CaNAC36 in peppers

Abstract [Objective] Pepper is the largest vegetable crop planted in China. With the land salinization becoming more and more serious, it is urgent to accelerate the functional study of key genes related to pepper salt tolerance. It is of great significance to strengthen the study of mechanisms underlying salt tolerance in peper to promote sustainable development of the pepper industry. [Methods] In the early stage of research, CaNAC36, a NAC transcription factor family gene related to salt stress response in peppers, was discovered. Based on this, the full-length gDNA and cDNA sequences of CaNAC36 were cloned using salttolerant pepper accession PI201224 and salt-sensitive pepper accession PI438643. The expression of CaNAC36 and its interacting genes in different tissues under salt stress was quantitatively analyzed by fluorescence, and the potential relationship between CaNAC36 and its interacting genes was further explored by bioinformatics analysis. [Results] The gDNA and cDNA sequence homology of CaNAC36 in salt-tolerant and salt-sensitive materials were 99.86% and 100%, respectively. CaNAC36 showed an up-expression in roots and stems in the salt-tolerant pepper, and a down-expression in roots and leaves in the salt-sensitive peper. Throgh analyzing the annotation information of 48 genes that may interact with CaNAC36, it was found that 14 genes were belonging to transmembrane proteins, transporter proteins, water porins, chloride channel proteins, detoxifying proteins, and other genes, which may have functional interaction with CaNAC36. Further analysis revealed that PI201224 and PI438643 exhibited significant differences in the expression of five related genes, inclding Capana08g002748, Capana00g004514, Capana09g000275, Capana07g001450, Capana02g001031, at different time points and tissues under salt stress. Meanwhile, the promoter region of CaNAC36 and five associated genes was found to contain a large number of stressrelated cis-acting elements. [Conclusion] CaNAC36 was an important regulatory gene in pepper in response to salt stress, and might interact with other genes to improve salt tolerance

An Elvitegravir Nanoformulation Crosses the Blood–Brain Barrier and Suppresses HIV-1 Replication in Microglia

Even with an efficient combination of antiretroviral therapy (ART), which significantly decreases viral load in human immunodeficiency virus type 1 (HIV-1)-positive individuals, the occurrence of HIV-1-associated neurocognitive disorders (HAND) still exists. Microglia have been shown to have a significant role in HIV-1 replication in the brain and in subsequent HAND pathogenesis. However, due to the limited ability of ART drugs to cross the blood–brain barrier (BBB) after systemic administration, in addition to efflux transporter expression on microglia, the efficacy of ART drugs for viral suppression in microglia is suboptimal. Previously, we developed novel poly (lactic-co-glycolic acid) (PLGA)-based elvitegravir nanoparticles (PLGA-EVG NPs), which showed improved BBB penetration in vitro and improved viral suppression in HIV-1-infected primary macrophages, after crossing an in vitro BBB model. Our objective in the current study was to evaluate the efficacy of our PLGA-EVG NPs in an important central nervous system (CNS) HIV-1 reservoir, i.e., microglia. In this study, we evaluated the cyto-compatibility of the PLGA-EVG NPs in microglia, using an XTT (2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide) assay and cellular morphology observation. We also studied the endocytosis pathway and the subcellular localization of PLGA NPs in microglia, using various endocytosis inhibitors and subcellular localization markers. We determined the ability of PLGA-EVG NPs to suppress HIV-1 replication in microglia, after crossing an in vitro BBB model. We also studied the drug levels in mouse plasma and brain tissue, using immunodeficient NOD scid gamma (NSG) mice, and performed a pilot study, to evaluate the efficacy of PLGA-EVG NPs on viral suppression in the CNS, using an HIV-1 encephalitic (HIVE) mouse model. From our results, the PLGA-EVG NPs showed ~100% biocompatibility with microglia, as compared to control cells. The internalization of PLGA NPs in microglia occurred through caveolae-/clathrin-mediated endocytosis. PLGA NPs can also escape from endo-lysosomal compartments and deliver the therapeutics to cells efficiently. More importantly, the PLGA-EVG NPs were able to show ~25% more viral suppression in HIV-1-infected human-monocyte-derived microglia-like cells after crossing the in vitro BBB compared to the EVG native drug, without altering BBB integrity. PLGA-EVG NPs also showed a ~two-fold higher level in mouse brain and a trend of decreasing CNS HIV-1 viral load in HIV-1-infected mice. Overall, these results help us to create a safe and efficient drug delivery method to target HIV-1 reservoirs in the CNS, for potential clinical use