Distinct temporal regulation of intrinsic and innate intracellular immunity to Herpes Simplex Virus type 1 (HSV-1) infection

Intrinsic and innate immunity play pivotal roles in limiting the replication of invading viral pathogens. Intrinsic immunity is constitutive and mediated by pre-existing host cell restriction factors (e.g., promyelocytic leukemia-nuclear body (PML-NB) constituent proteins) which directly confer antiviral properties. On the other hand, innate immunity is inducible and upregulated in response to infection. Pattern recognition receptors (PRRs) (e.g., interferon gamma inducible protein 16 (IFI16)) sense pathogen-associated molecular patterns (PAMPs) and induce downstream signaling cascades leading to the induction of Interferon-stimulated gene (ISG) products that confer antiviral properties. These two arms of immunity represent the first line of intracellular defense to HSV-1 infection. Indeed, rapid recruitment of intrinsic and innate immune factors to viral DNA (vDNA) has a significant bearing on the outcome of infection. However, the spatial and temporal regulation of this recruitment remains poorly defined due to the technical challenges associated with vDNA detection at multiplicities of infection (MOI) that do not saturate intrinsic host factors. Utilizing 5-Ethynyl-2’-deoxyuridine (EdU) labeling of HSV-1 DNA in combination with click chemistry, we directly visualized input viral genomes under low MOI conditions (MOI of ≤ 3 PFU/cell) at 30-90 minutes post-addition of virus (mpi). This protocol is sensitive, specific, and compatible with indirect immunofluorescence (IF) staining protocols, providing a valuable assay to investigate the temporal recruitment of immune regulators to infecting vDNA. Upon entry of vDNA into the nucleus, PML-NB associated restriction factors (e.g., PML, SP100, and Daxx) were rapidly recruited to infecting viral genome foci. This process occurred in a PML-dependent manner and led to genome entrapment and silencing within PML-NBs. Interestingly, genome entrapment was observed during both wild-type (WT) and ICP0-null mutant (ΔICP0) HSV-1 infection. During WT HSV-1 infection, ICP0 induced PML degradation and the dispersal of PML-NB restriction factors, highlighting the importance of ICP0 to release viral genomes entrapped within PML-NBs to stimulate the onset of lytic HSV-1 replication. During ΔICP0 HSV-1 infection, vDNA remained stably entrapped within PML-NBs leading to a repression in viral gene expression and a restriction in plaque formation. Importantly, IFI16 was not stably recruited to vDNA entrapped within PML-NBs, and ISG expression was not induced under low MOI conditions that do not saturate PML-NB intrinsic host defenses. These data demonstrate that vDNA entry into the nucleus alone is not sufficient to stimulate the induction of innate immunity. Saturation of intrinsic host defenses under high MOI conditions stimulated the stable recruitment of IFI16 to infecting viral genomes, and induced ISG expression in a PML-, IFI16-, and Janus-associated kinase (JAK)-dependent manner. The induction of this innate immune response was dependent on the onset of vDNA replication, as treatment of the infected cell monolayers with phosphonoacetic acid (PAA), a vDNA polymerase inhibitor, inhibited ISG induction in a dose-dependent manner. Unlike PML depletion, inhibition of JAK signaling failed to relieve the plaque formation defect of ΔICP0 HSV-1, but instead significantly enhanced virus yields. Collectively, these data, for the first time, demonstrate a temporal and sequential induction of intrinsic and innate immunity during HSV-1 infection. Intrinsic immunity is induced within minutes of nuclear infection to restrict the initiation of viral gene transcription and the onset of lytic replication. Escape from this intrinsic repression and initiation of vDNA replication, which takes several hours, triggers the induction of innate immunity. ISG products establish an antiviral state within infected and neighboring uninfected cells to constrict viral propagation and limit the spread of infection. We identify dual roles for PML in the regulation of intrinsic and innate immunity to HSV-1 infection. However, these host defenses are counteracted by the viral ubiquitin ligase ICP0, which targets PML for degradation to promote vDNA release from PML-NBs in order to evade intrinsic viral genome silencing from the onset of nuclear infection

Synthesis and Study of Modified Nanostructure Porous Silicon Layers for Chemical Gas Sensing

In this work, We prepared a modified nanostructure porous silicon (PS) layers for effective chemical gas sensing. Nanopore covered microporous silicon gas sensor has been fabricated using electrochemical etching in an HF acid and ethanol solution. A porous silicon (PS) surface has been modified using selective depositions formed from metal to enhance the response to Sensing of CO2. (PS) has been interest for gas sensing because of the exceptional gathering of importent features. By setting the process parameters,the porosity, pore size, and the morphology can be modifid and practically controlled. The modified porous silicon layers were characterized using different techniques such as scanning electron microscopy(SEM)and a series of electrical characterizations to study the structures in the contact of the carbon dioxide was achieved

Optimising data processing for nanodiamond based relaxometry

The nitrogen-vacancy (NV) center in diamond is a powerful and versatile quantum sensor for diverse quantities. In particular, relaxometry (or T1), allows to detect magnetic noise at the nanoscale. While increasing the number of NV centers in a nanodiamond allows to collect more signal, a standardized method to extract information from relaxometry experiments of such NV ensembles is still missing. In this article, we use T1 relaxation curves acquired at different concentrations of gadolinium ions to calibrate and optimize the entire data processing flow, from the acquired raw data to the extracted T1. In particular, we use a bootstrap to derive a signal to noise ratio (SNR) that can be quantitatively compared from one method to another. At first, T1 curves are extracted from photoluminescence pulses. We compare integrating their signal through an optimized window as performed conventionally, to fitting a known function on it. Fitting the decaying T1 curves allows to obtain the relevant T1 value. We compared here the three most commonly used fit models that are, single, bi, and stretched-exponential. We finally investigated the effect of the bootstrap itself on the precision of the result as well as the use of a rolling window to allows time-resolution.Comment: With Advanced Quantum Technologies (26/04/2023) / Supplementary information included / 15 pages + 1SI, 6 figures + 1SI, 58 reference

A Candida albicans early stage biofilm detachment event in rich medium

<p>Abstract</p> <p>Background</p> <p>Dispersal from <it>Candida albicans </it>biofilms that colonize catheters is implicated as a primary factor in the link between contaminated catheters and life threatening blood stream infections (BSI). Appropriate in vitro <it>C. albicans </it>biofilm models are needed to probe factors that induce detachment events.</p> <p>Results</p> <p>Using a flow through system to culture <it>C. albicans </it>biofilms we characterized a detachment process which culminates in dissociation of an entire early stage biofilm from a silicone elastomer surface. We analyzed the transcriptome response at time points that bracketed an abrupt transition in which a strong adhesive association with the surface is weakened in the initial stages of the process, and also compared batch and biofilm cultures at relevant time points. K means analysis of the time course array data revealed categories of genes with similar patterns of expression that were associated with adhesion, biofilm formation and glycoprotein biosynthesis. Compared to batch cultures the biofilm showed a pattern of expression of metabolic genes that was similar to the <it>C. albicans </it>response to hypoxia. However, the loss of strong adhesion was not obviously influenced by either the availability of oxygen in the medium or at the silicone elastomer surface. The detachment phenotype of mutant strains in which selected genes were either deleted or overexpressed was characterized. The microarray data indicated that changes associated with the detachment process were complex and, consistent with this assessment, we were unable to demonstrate that transcriptional regulation of any single gene was essential for loss of the strong adhesive association.</p> <p>Conclusion</p> <p>The massive dispersal of the early stage biofilm from a biomaterial surface that we observed is not orchestrated at the level of transcriptional regulation in an obvious manner, or is only regulated at this level by a small subpopulation of cells that mediate adhesion to the surface.</p

Titanium-Base Nanostructure Coatings for AISI M52 Tool Steel by Gas-Phase Mix Process

Deposition of Multicomponent hard coatings (Ti-B-N-C) on the molybdenum high-speed tool steel (AISI M52) has been achieved by mixed vapor deposition technique to improve the mechanical properties of the surface. In this technique the coating materials that were supplied in the gas phase were produced from powders that vaporized by thermal energy (that is, PVD- Reactive Evaporation Process), while the reactor that used to deposit Ti-Base coatings is hot-wall chemical vapor deposition (HWCVD) system equipment. This combination results in technical and financial advantages. The structure of deposited Ti-Base hard film was characterized by XRD technique, Scanning Electron Microscopy (SEM), and Energy Dispersive X-ray spectroscopy (EDX). Mechanical characterization of the hard films has been performed by using Vickers micro hardness tester and The Ball-on-disk wear tests. With different reactive gas flow rates that ranged from 500 to 3000Sccm(standard Cubic Centimeter per Minute), the film showed amorphous matrix with crystalline Nano fibers of Ti-B phase which led to achieve higher hardness of 2051HV and better wear resistance with relatively good COF values of 0.61 than the uncoated tools

Improve the Process of Enhancing Oil Recovery (EOR) by Applying Nanomagnetic Cobalt Ferrite Nanoparticles

In this paper we reported nano-crystalline cobalt ferrite powders were synthesized using co-precipitation method at 600 °C, 700 °C and 800 °C. The structural, morphological and magnetic properties of the powders were investigated by x-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, transmission electron microscopy (TEM) and vibrating sample magnetometer (VSM). Infrared spectral analysis data between 200 and 1000 cm-1 defined the intrinsic cation vibrations of the characteristic spinel structure system. The saturation magnetization (Ms) and coercivity (Hc) of the CoFe2O4 were found to be in the range of 94-33 emu/g, which is still in the range of hard ferrite. The observed variation in saturation magnetization, coercivity and remanence magnetization as a function of increasing the temperature and grain size of samples. From this point of view, nano-scale size of nanoparticles makes them efficient for using in borehole stability maintaining for enhancing oil and gas recovery efficiency improvement. The large value of magnetic pressure (-2.95699) are expected to be useful in oil recovery applications. It has also been found that the choice of nanoparticles for application in oil recovery depends on nature, magnetic and electric properties of the reservoir rock

Performance and Combustion Characteristics of an SI Engine Fueled With Fusel Oil-Gasoline at Different Water Content

The main objective of the current study is to investigate the impacts of fusel oil-gasoline blends with different water content on performance and combustion characteristics of an SI engine. The water content of fusel oil is reduced by employing rotary extractor method. The parameters examined in the current study are the effects of test fuels upon engine performance criteria, fuel combustion, location of CA50 and coefficient of variation of indicated mean effective pressure (COVIMEP). The experiments are conducted in a SI engine with two blending ratio of gasoline-fusel oil (FBWE10 and FAWE10) in addition to pure gasoline as a standard fuel under different engine speeds and constant 45% of WOT engine load. The study results reveal that engine power, torque, and brake thermal efficiency have slightly improved with fusel oil-gasoline blends that were obvious clearly after reduction of water content. On the other hand, though the brake specific fuel consumption of FAWE10 slightly improve compared to FBWE10 but still higher than gasoline. Furthermore, the peak in-cylinder pressure, rate of heat release, and rate of pressure rise enhanced with fusel oil-gasoline blends. The (CA 0–10) and (CA 10–90) duration could be simultaneously decreased by reducing the water content of fusel oil, all of that are helpful to improve the stability of combustion and thought to be reasons for the reduction in COVIMEP. As a conclusion, the reduction of fusel oil water content has played positively to enhance the combustion and performance characteristics of the fuel blend and result in more stable engine operation

Retarding of Precipitation Hardening of Al-Cu Alloy by Cadimium Addition

The aim of the present work is to prepare a set of Al-Cu alloys with slow precipitation hardening by aging. Present work results can be utilized during the importing or exporting of such alloys at solution treatment conditions. In order to establish this aim, a set of Al-4%Cu alloys have been prepared in the laboratory to investigate the effect of Cd-addition with the percentage varied from (0.1, 0.3 and 0.5) weight percentage. The prepared alloys with different Cd additions were subjected to a natural aging up to 60 days after solution treatment. As a result, the alloys showed an obvious retarding in the aging kinetics as the percentage of Cdadditions increasing. This retarding was found to be due to the trapping of the vacancies that resulted during the quenching step that resulted in delays of Gunier- Preston zones precipitation. SEM has been used in monitoring of microstructure as well as optical microscop

A COMPARATIVE STUDY OF QUALITY CONTROL TESTING OF MEFENAMIC ACID TABLETS IN IRAQ

Objective: This research was performed to assess the quality of different marketed tablets having mefenamic acid (500 mg). The selected tablets are produced by numerous companies and presented in the Iraqi pharmaceutical marketplace. Methods: Different batches of mefenamic acid conventional tablets were exposed for several tests of quality control. These evaluation tests include hardness, weight variation, friability, disintegration time, drug content, and drug dissolution profile. The properties of these quality tests were made conferring to the specification of USP-pharmacopeia. Results: The data of this study indicate that each tablet of mefenamic acid batches conformed to the requirement of USP pharmacopeia, the hardness was (6.87-8.06 Kg/cm2), and the drug content results were (90.666-99.214%) within USP limitation. The data of disintegration time and weight uniformity were agreeable with pharmacopeia and the in vitro release assay showed that the release of each mefenamic acid marketed tablet was highest than (80 %) in 45 min, which reproducing compliance with the USP pharmacopeia's limitation. Conclusion: From this study, it was proved that all of the marketed brands of mefenamic acid tablets meet the standard character in the USP pharmacopeia for in vitro quality control tests

On Involving Infrastructure Topics in the Civil Engineering Department Curriculum-Survey of Graduate Students Views

In this work, a survey on the Ishik-University graduate students’ level of awareness of the importance of infrastructures on the community, on their views on the curriculum versus infrastructures’ topics, and the most important topics found better for their future work. Our goal is to prepare our students to be future engineers educated with the concepts of infrastructures and how to improve and maintain these facilities. Students have been given a chance to evaluate current curriculum with particular focus on the infrastructure topics. Students have shown their high level of awareness, and gave their opinion on the curriculum and infrastructure as shown in tables and graphs. Conclusions and recommendations for the civil engineering department in Ishik University are drawn. It is the aim of the authors to further improve the curriculum of the civil engineering department of Ishik University, particularly aiming at providing our graduates a better understanding of infrastructures