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Porcine cytomegalovirus (PCMV) is widely distributed in pigs and difficult to detect due to latency. PCMV infection of source pigs was associated with early graft failure after cardiac and renal xenotransplantation into nonhuman primates. Importantly, PCMV infection of the first genetically modified pig heart into a human may have contributed to the reduced survival of the patient. Sensitive and reliable assays for detection of latent PCMV infection are thus indispensable. Here, we report the development of five peptide-induced rabbit antisera specific for PCMV glycoprotein B (gB) and their validation for detection of PCMV in infected pig fallopian tube (PFT) cells by immunofluorescence and electron microscopy (EM). The anti-gB antibodies were also used for detection by Western blot analysis of PCMV purified from the supernatant of infected PFT cells. Sera of infected versus non-infected pigs have been compared. In parallel, PCMV viral load in blood samples of the animals was quantified by a novel highly sensitive nested-PCR and qPCR assay. A combination of four partly overlapping peptides from the gB C-terminus was used to establish a diagnostic ELISA for PCMV gB specific pig antibodies which is able to differentiate infected from non-infected animals and to quantify maternal antibodies in neonates. The combination of a highly sensitive nested PCR for direct virus detection with a sensitive peptide-based ELISA detecting anti-PCMV gB-antibodies, supplemented by Western blot analysis and/or immunohistochemistry for virus detection will reliably differentiate pigs with active infection, latently infected pigs, and non-infected pigs. It may significantly improve the virologic safety of xenotransplantation.
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Infecções por Citomegalovirus , Citomegalovirus , Feminino , Animais , Suínos , Humanos , Coelhos , Citomegalovirus/genética , Transplante Heterólogo , Infecções por Citomegalovirus/diagnóstico , Reação em Cadeia da Polimerase , PeptídeosRESUMO
We present a tunable, hybrid waveguide-fiber optical parametric oscillator (OPO) synchronously pumped by an ultra-fast fiber laser exploiting four-wave mixing (FWM) generated in silicon nitride waveguides. Parametric oscillation results in a 35 dB enhancement of the idler spectral power density in comparison to spontaneous FWM, with the ability of wide wavelength tuning over 86 nm in the O-band. Measurements of the oscillation threshold and the efficiency of the feedback loop reveal how an integration of the OPO on a single silicon nitride chip can be accomplished at standard repetition rates of pump lasers in the order of 100 MHz.
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We present tunable waveguide-based optical parametric amplification by four-wave mixing (FWM) in silicon nitride waveguides, with the potential to be set up as an all-integrated device, for narrowband coherent anti-Stokes Raman scattering (CARS) imaging. Signal and idler pulses are generated via FWM with only 3 nJ pump pulse energy and stimulated by using only 4 mW of a continuous-wave seed source, resulting in a 35 dB enhancement of the idler spectral power density in comparison to spontaneous FWM. By using waveguides with different widths and tuning the wavelength of the signal wave seed, idler wavelengths covering the spectral region from 1.1 µm up to 1.6 µm can be generated. The versatility of the chip-based FWM light source is demonstrated by acquiring CARS images.
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Extending the cavity length of diode lasers with feedback from Bragg structures and ring resonators is highly effective for obtaining ultra-narrow laser linewidths. However, cavity length extension also decreases the free-spectral range of the cavity. This reduces the wavelength range of continuous laser tuning that can be achieved with a given phase shift of an intracavity phase tuning element. We present a method that increases the range of continuous tuning to that of a short equivalent laser cavity, while maintaining the ultra-narrow linewidth of a long cavity. Using a single-frequency hybrid integrated InP-Si3N4 diode laser with 120 nm coverage around 1540 nm, with a maximum output of 24 mW and lowest intrinsic linewidth of 2.2 kHz, we demonstrate a six-fold increased continuous and mode-hop-free tuning range of 0.22 nm (28 GHz) as compared to the free-spectral range of the laser cavity.
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We demonstrate a hybrid integrated and widely tunable diode laser with an intrinsic linewidth as narrow as 40 Hz, achieved with a single roundtrip through a low-loss feedback circuit that extends the cavity length to 0.5 meter on a chip. Employing solely dielectrics for single-roundtrip, single-mode resolved feedback filtering enables linewidth narrowing with increasing laser power, without limitations through nonlinear loss. We achieve single-frequency oscillation with up to 23 mW fiber coupled output power, 70-nm wide spectral coverage in the 1.55 µm wavelength range with 3 mW output and obtain more than 60 dB side mode suppression. Such properties and options for further linewidth narrowing render the approach of high interest for direct integration in photonic circuits serving microwave photonics, coherent communications, sensing and metrology with highest resolution.
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We present a light source for coherent anti-Stokes Raman scattering (CARS) based on broadband spontaneous four-wave mixing, with the potential to be further integrated. By using 7 mm long silicon nitride waveguides, which offer tight mode confinement and a high nonlinear refractive index coefficient, broadband signal and idler pulses were generated with 4 nJ of input pulse energy. In comparison to fiber-based experiments, the input energy and the waveguide length were reduced by two orders of magnitude, respectively. The idler and residual pump pulses were used for CARS measurements, enabling chemically selective and label-free spectroscopy over the entire fingerprint region, with an ultrafast fiber-based pump source at 1033 nm wavelength. The presented simple light source paves the path towards cost-effective, integrated lab-on-a-chip CARS applications.
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Tetraspanins are master organizers of the cell membrane. Recent evidence suggests that tetraspanins themselves may become crowded by virus particles and that these crowds/aggregates co-internalize with the viral particles. Using microscopy, we studied human papillomavirus (HPV) type 16-dependent aggregates on the cell surface of tetraspanin overexpressing keratinocytes. We find that aggregates are (1) rich in at least two different tetraspanins, (2) three-dimensional architectures extending up to several micrometers into the cell, and (3) decorated intracellularly by filamentous actin. Moreover, in cells not overexpressing tetraspanins, we note that obscurin-like protein 1 (OBSL1), which is thought to be a cytoskeletal adaptor, associates with filamentous actin. We speculate that HPV contact with the cell membrane could trigger the formation of a large tetraspanin web. This web may couple the virus contact site to the intracellular endocytic actin machinery, possibly involving the cytoskeletal adaptor protein OBSL1. Functionally, such a tetraspanin web could serve as a virus entry platform, which is co-internalized with the virus particle.
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Actinas/fisiologia , Proteínas do Citoesqueleto/fisiologia , Papillomavirus Humano 16/fisiologia , Tetraspanina 24/fisiologia , Tetraspanina 30/fisiologia , Endocitose , Células HaCaT/virologia , Células HeLa/ultraestrutura , Células HeLa/virologia , Células Hep G2/virologia , Humanos , Microscopia Confocal , Microscopia Eletrônica , Infecções por Papillomavirus/virologia , Plaquinas/fisiologia , Vírion/fisiologia , Vírion/ultraestrutura , Internalização do VírusRESUMO
The infection of human transplant recipients by porcine endogenous retrovirus (PERV) is a safety issue for xenotransplantation (XTx). CRISPR/Cas9 technology has enabled the generation of pigs free of functional PERVs, and the susceptibility of these animals to reinfection by PERVs remains unclear. To assess virological safety, we characterized a cell line in which PERVs have been inactivated by CRISPR/Cas9 (PK15 clone 15) for its susceptibility to infectious PERV. First, basal expression of PERV pol, the porcine PERV-A receptor (POPAR), and reverse transcriptase (RT) activity of PERV were determined. PK15 clone 15 cells were inoculated with PERV and monitored post infection for virus expression and RT activity. Particles were visualized by electron microscopy. Our data show that PK15 clone 15 cells still produce viral proteins that assemble to produce impaired viral particles. These virions have an irregular morphology that diverges from that of mature wild type. The particles are no longer infectious when tested in a downstream infection assay using supernatants of PK15 clone 15 cells to infect susceptible swine testis-IOWA (ST-IOWA) cells. The expression of POPAR was quantified to exclude the possibility that lack of susceptibility to reinfection, for PERV-A, is caused by absence of viral host receptor(s). PK15 and PK15 clone 15 cells do, in fact, express POPAR equally. PERV RT inactivation mediated by CRISPR/Cas9 does not compromise virus assembly but affects virion structure and proviral integration. The constitutive virion production seems to maintain cellular resistance to superinfection and possibly indicates a protective side effect of this specific CRISPR/Cas9 mediated RT inactivation.
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Sistemas CRISPR-Cas/fisiologia , Retrovirus Endógenos/patogenicidade , Provírus/patogenicidade , Porco Miniatura/virologia , Animais , Linhagem Celular , Humanos , Suínos , Transplante Heterólogo/efeitos adversosRESUMO
BACKGROUND: Platelet concentrates play an important role in transfusion medicine. Their short lifespan and lack of robustness require efforts to ensure adequate product quality. In this study, we compared the in vitro quality of the main concentrate types, pooled platelet concentrate (PPC) from whole blood donations, and platelet concentrate from single-donor apheresis (APC). METHODS: Twenty PPCs and 20 APCs prepared in plasma were analyzed on days 2, 4, and 7 of storage. Variables related to metabolism, degranulation, platelet aggregation, P-selectin expression, and annexin V binding were analyzed. Morphology was assessed by transmission electron microscopy of ultrathin sections. A microfluidic device was applied to test the effects of shear stress on platelet function. RESULTS: The metabolic parameters indicated stable storage conditions throughout the 7-day period. The resting discoid form was the prevailing morphology on days 2 and 4 in the PPCs and APCs. Chemokine release and receptor shedding of soluble P-selectin and soluble CD40L equally increased in PPCs and APCs. Aggregation responses to ADP and collagen were heterogeneous, with marked losses in collagen responsiveness on day 4 in individual concentrates. Baseline expression of P-selectin in PPCs and APCs was low, and inducibility of P-selectin was well preserved until day 4. Under shear stress, equal adhesiveness and stability were found with platelets from PPCs and APCs. CONCLUSIONS: Platelets from PPCs and APCs showed similar in vitro function and stability parameters. However, platelet concentrates presented a high variability and individual concentrates an impaired functional capability. Identifying the factors contributing to this would help increase product reliability.
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We theoretically investigate the use of Rayleigh surface acoustic waves (SAWs) for refractive index modulation in optical waveguides consisting of amorphous dielectrics. Considering low-loss Si3N4 waveguides with a standard core cross-section of 4.4×0.03 µm2 size, buried 8-µm deep in a SiO2 cladding, we compare surface acoustic wave generation in various different geometries via a piezo-active, lead zirconate titanate film placed on top of the surface and driven via an interdigitized transducer (IDT). Using numerical solutions of the acoustic and optical wave equations, we determine the strain distribution of the SAW under resonant excitation. From the overlap of the acoustic strain field with the optical mode field, we calculate and maximize the attainable amplitude of index modulation in the waveguide. For the example of a near-infrared wavelength of 840 nm, a maximum shift in relative effective refractive index of 0.7x10-3 was obtained for TE polarized light, using an IDT period of 30-35 µm, a film thickness of 2.5-3.5 µm, and an IDT voltage of 10 V. For these parameters, the resonant frequency is in the range of 70-85 MHz. The maximum shift increases to 1.2x10-3, with a corresponding resonant frequency of 87 MHz, when the height of the cladding above the core is reduced to 3 µm. The relative index change is about 300 times higher than in previous work based on non-resonant proximity piezo-actuation, and the modulation frequency is about 200 times higher. Exploiting the maximum relative index change of 1.2×10-3 in a low-loss, balanced Mach-Zehnder modulator should allow full-contrast modulation in devices as short as 120 µm (half-wave voltage length product = 0.24 Vcm).
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We present an integrated hybrid semiconductor-dielectric (InP-Si3N4) waveguide laser that generates frequency combs at a wavelength around 1.5 µm with a record-low intrinsic optical linewidth of 34 kHz. This is achieved by extending the cavity photon lifetime using a low-loss dielectric waveguide circuit. In our experimental demonstration, the on-chip, effective optical path length of the laser cavity is extended to 6 cm. The resulting linewidth narrowing shows the high potential of on-chip, highly coherent frequency combs with direct electrical pumping, based on hybrid and heterogeneous integrated circuits making use of low-loss dielectric waveguides.
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The development of large-scale optical quantum information processing circuits ground on the stability and reconfigurability enabled by integrated photonics. We demonstrate a reconfigurable 8×8 integrated linear optical network based on silicon nitride waveguides for quantum information processing. Our processor implements a novel optical architecture enabling any arbitrary linear transformation and constitutes the largest programmable circuit reported so far on this platform. We validate a variety of photonic quantum information processing primitives, in the form of Hong-Ou-Mandel interference, bosonic coalescence/anti-coalescence and high-dimensional single-photon quantum gates. We achieve fidelities that clearly demonstrate the promising future for large-scale photonic quantum information processing using low-loss silicon nitride.
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We demonstrate the potential of all-optical switches in integrated waveguides based on intermodal cross-phase modulation between transverse modes. For this purpose, the differential phase between two transverse modes of a probe beam was altered by cross-phase modulation with a control beam propagating only in the fundamental mode. A switching behavior was accomplished by spatially filtering the resulting multimode interference of the probe modes, which changed depending on the control beam power. All-optical switching with a contrast of 82% at 1280 nm over a frequency range of 4.4 THz at 1.6 nJ was achieved, representing an improvement of the product of necessary power and waveguide length by a factor of nearly 2000 compared to similar experiments in graded-index fibers. Additionally, we show that the center wavelength of the switch can be tailored by changing the cross-sectional geometry of the waveguide or the involved probe modes.
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The suppression of Raman scattering is of high interest for the achievement of sub-diffraction-limited resolution in Raman scattering spectroscopy and microscopy. We present density matrix calculations of the suppression of spontaneous Raman scattering via ground state depletion in a level system based on the molecule tris(bipyridine)ruthenium(ii). This particular molecule has been earlier used for an experimental demonstration of the suppression of spontaneous Raman scattering, allowing us to successfully verify the validity of our numerical calculations by a comparison to the experimental results. We investigate the required level of detail of the molecule model as well as the influence of certain molecule and pulse parameters on the Raman scattering suppression. It was found that pulses with a duration longer than the lifetime of the electronic states allow for a high suppression of the Raman scattering. Pulses shorter than the coherence lifetime between the ground state and electronic states lead to a similarly high suppression but also accomplish the suppression with more than one order of magnitude lower pulse energy fluence. Additionally, using a laser wavelength that is in resonance with one of the electronic transitions of the sample should allow suppressing the Raman scattering with four to six orders of magnitude lower pulse energy fluence.
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Hepatitis C virus (HCV) particles are described as lipoviroparticles which are released similarly to very-low-density lipoproteins (VLDLs). However, the release mechanism is still poorly understood; the canonical endoplasmic reticulum-Golgi intermediate compartment (ERGIC) pathway as well as endosome-dependent release has been proposed. Recently, the role of exosomes in the transmission of HCV has been reported. Only a minor fraction of the de novo-synthesized lipoviroparticles is released by the infected cell. To investigate the relevance of multivesicular bodies (MVBs) for viral morphogenesis and release, the MVB inhibitor U18666A was used. Intracellular trafficking was analyzed by confocal microscopy and electron microscopy. Moreover, an mCherry-tagged HCV variant was used. Conditions were established that enable U18666A-dependent inhibition of MVBs without affecting viral replication. Under these conditions, significant inhibition of the HCV release was observed. The assembly of viral particles is not affected. In U18666A-treated cells, intact infectious viral particles accumulate in CD63-positive exosomal structures and large dysfunctional lysosomal structures (multilamellar bodies). These retained particles possess a lower density, reflecting a misloading with lipids. Our data indicate that at least a fraction of HCV particles leaves the cell via the endosomal pathway. Endosomes facilitate the sorting of HCV particles for release or degradation. IMPORTANCE: There are still a variety of open questions regarding morphogenesis and release of hepatitis C virus. The HCV-infected cell produces significant more viral particles that are released, raising the question about the fate of the nonreleased particles. Moreover, the relevance of the endosomal pathway for the release of HCV is under debate. Use of the MVB (multivesicular body) inhibitor U18666A enabled a detailed analysis of the impact of MVBs for viral morphogenesis and release. It was revealed that infectious, fully assembled HCV particles are either MVB-dependently released or intracellularly degraded by the lysosome. Our data indicate that at least a fraction of HCV particles leaves the cell via the endosomal pathway independent from the constitutive secretory pathway. Our study describes a so-far-unprecedented cross talk between two pathways regulating on the one hand the release of infectious viral particles and on the other hand the intracellular degradation of nonreleased particles.
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Androstenos/farmacologia , Anticolesterolemiantes/farmacologia , Exossomos/efeitos dos fármacos , Hepacivirus/efeitos dos fármacos , Hepatócitos/efeitos dos fármacos , Liberação de Vírus/efeitos dos fármacos , Animais , Transporte Biológico/efeitos dos fármacos , Linhagem Celular Tumoral , Colesterol/metabolismo , Exossomos/ultraestrutura , Exossomos/virologia , Expressão Gênica , Genes Reporter , Hepacivirus/fisiologia , Hepacivirus/ultraestrutura , Hepatócitos/ultraestrutura , Hepatócitos/virologia , Humanos , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Corpos Multivesiculares/efeitos dos fármacos , Corpos Multivesiculares/ultraestrutura , Corpos Multivesiculares/virologia , Vírion/efeitos dos fármacos , Vírion/fisiologia , Vírion/ultraestrutura , Montagem de Vírus/fisiologia , Proteína Vermelha FluorescenteRESUMO
All Staphylococcus aureus genomes contain a genomic island, which is termed νSaα and characterized by two clusters of tandem repeat sequences, i.e. the exotoxin (set) and 'lipoprotein-like' genes (lpl). Based on their structural similarities the νSaα islands have been classified as type I to IV. The genomes of highly pathogenic and particularly epidemic S. aureus strains (USA300, N315, Mu50, NCTC8325, Newman, COL, JH1 or JH9) belonging to the clonal complexes CC5 and CC8 bear a type I νSaα island. Since the contribution of the lpl gene cluster encoded in the νSaα island to virulence is unclear to date, we deleted the entire lpl gene cluster in S. aureus USA300. The results showed that the mutant was deficient in the stimulation of pro-inflammatory cytokines in human monocytes, macrophages and keratinocytes. Purified lipoprotein Lpl1 was further shown to elicit a TLR2-dependent response. Furthermore, heterologous expression of the USA300 lpl cluster in other S. aureus strains enhanced their immune stimulatory activity. Most importantly, the lpl cluster contributed to invasion of S. aureus into human keratinocytes and mouse skin and the non-invasive S. carnosus expressing the lpl gene cluster became invasive. Additionally, in a murine kidney abscess model the bacterial burden in the kidneys was higher in wild type than in mutant mice. In this infection model the lpl cluster, thus, contributes to virulence. The present report is one of the first studies addressing the role of the νSaα encoded lpl gene cluster in staphylococcal virulence. The finding that the lpl gene cluster contributes to internalization into non-professional antigen presenting cells such as keratinocytes highlights the lpl as a new cell surface component that triggers host cell invasion by S. aureus. Increased invasion in murine skin and an increased bacterial burden in a murine kidney abscess model suggest that the lpl gene cluster serves as an important virulence factor.
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Ilhas Genômicas/genética , Infecções Estafilocócicas/genética , Staphylococcus aureus/genética , Staphylococcus aureus/patogenicidade , Virulência/genética , Animais , Células Cultivadas , DNA Bacteriano/genética , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Feminino , Técnicas de Inativação de Genes , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica de Transmissão , Microscopia de Fluorescência , Fatores de Virulência/genéticaRESUMO
We demonstrate supercontinuum generation in stoichiometric silicon nitride (Si3N4 in SiO2) integrated optical waveguides, pumped at telecommunication wavelengths. The pump laser is a mode-locked erbium fiber laser at a wavelength of 1.56 µm with a pulse duration of 120 fs. With a waveguide-internal pulse energy of 1.4 nJ and a waveguide with 1.0 µm × 0.9 µm cross section, designed for anomalous dispersion across the 1500 nm telecommunication range, the output spectrum extends from the visible, at around 526 nm, up to the mid-infrared, at least to 2.6 µm, the instrumental limit of our detection. This output spans more than 2.2 octaves (454 THz at the -30 dB level). The measured output spectra agree well with theoretical modeling based on the generalized nonlinear Schrödinger equation. The infrared part of the supercontinuum spectra shifts progressively towards the mid-infrared, well beyond 2.6 µm, by increasing the width of the waveguides.
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BACKGROUND: Zika virus (ZIKV) is an emerging mosquito-borne Flavivirus of major public health concern. The potential for ZIKV transmission by blood transfusion has been demonstrated; however, inactivation or removal of ZIKV during the manufacture of plasma-derived medicinal products has not been specifically investigated. STUDY DESIGN AND METHODS: Inactivation of ZIKV by pasteurization and solvent/detergent (S/D) treatment was investigated by spiking high-titer ZIKV stocks into human serum albumin and applying either heat or adding different mixtures of S/D reagents and assaying for infectious virus particles. Removal of ZIKV was evaluated using filters of differing pore sizes (75, 40, 35, and 19 nm), assaying for infectious virus and RNA. Electron microscopy was performed to determine the size of ZIKV particles. Neutralization of virus infectivity by immunoglobulins was investigated. RESULTS: ZIKV was effectively and rapidly inactivated by liquid heat treatment as well as by various mixtures of S/D reagents with reduction factors more than 4 log, in each case. Effective reduction of ZIKV infectivity was demonstrated for virus filtration for filters with average pore sizes of not more than 40 nm, although a significant proportion of virus RNA was detected in the 40- to 35-nm filtrates likely due to the presence of subviral particles observed by electron microscopy. None of the immunoglobulin preparations investigated neutralized ZIKV infectivity. CONCLUSIONS: Pasteurization and S/D treatment very rapidly inactivated ZIKV and filters with a pore size of not more than 40 nm removed all infectious ZIKV, demonstrating the effectiveness of these virus reduction strategies used during the manufacture of plasma-derived medicinal products.
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Desinfecção/métodos , Plasma/virologia , Inativação de Vírus , Infecção por Zika virus/prevenção & controle , Zika virus , Animais , Anticorpos Antivirais/química , Chlorocebus aethiops , Feminino , Humanos , Masculino , Plasma/química , Ultrafiltração/métodos , Células VeroRESUMO
UNLABELLED: In addition to infectious viral particles, hepatitis B virus-replicating cells secrete large amounts of subviral particles assembled by the surface proteins, but lacking any capsid and genome. Subviral particles form spheres (22-nm particles) and filaments. Filaments contain a much larger amount of the large surface protein (LHBs) compared to spheres. Spheres are released via the constitutive secretory pathway, while viral particles are ESCRT-dependently released via multivesicular bodies (MVBs). The interaction of virions with the ESCRT machinery is mediated by α-taxilin that connects the viral surface protein LHBs with the ESCRT component tsg101. Since filaments in contrast to spheres contain a significant amount of LHBs, it is unclear whether filaments are released like spheres or like virions. To study the release of subviral particles in the absence of virion formation, a core-deficient HBV mutant was generated. Confocal microscopy, immune electron microscopy of ultrathin sections and isolation of MVBs revealed that filaments enter MVBs. Inhibition of MVB biogenesis by the small-molecule inhibitor U18666A or inhibition of ESCRT functionality by coexpression of transdominant negative mutants (Vps4A, Vps4B, and CHMP3) abolishes the release of filaments while the secretion of spheres is not affected. These data indicate that in contrast to spheres which are secreted via the secretory pathway, filaments are released via ESCRT/MVB pathway like infectious viral particles. IMPORTANCE: This study revises the current model describing the release of subviral particles by showing that in contrast to spheres, which are secreted via the secretory pathway, filaments are released via the ESCRT/MVB pathway like infectious viral particles. These data significantly contribute to a better understanding of the viral morphogenesis and might be helpful for the design of novel antiviral strategies.
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Proteínas de Ligação a DNA/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Vírus da Hepatite B/metabolismo , Corpos Multivesiculares/metabolismo , Fatores de Transcrição/metabolismo , Liberação de Vírus/fisiologia , Androstenos/farmacologia , Linhagem Celular Tumoral , Citoesqueleto/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte/antagonistas & inibidores , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Células Hep G2 , Vírus da Hepatite B/genética , Hepatócitos/virologia , Humanos , Microscopia Confocal , Microscopia Eletrônica , Corpos Multivesiculares/efeitos dos fármacos , Proteínas de Transporte Vesicular/metabolismo , Proteínas do Core Viral/deficiência , Proteínas do Core Viral/genéticaRESUMO
We report on the first experimental demonstration of the suppression of spontaneous Raman scattering via ground state depletion. The concept of Raman suppression can be used to achieve sub-diffraction-limited resolution in label-free microscopy by exploiting spatially selective signal suppression when imaging a sample with a combination of Gaussian- and donut-shaped beams and reconstructing a resolution-enhanced image from this data. Using a nanosecond pulsed laser source with an emission wavelength of 355 nm, the ground state of tris(bipyridine)ruthenium(II) molecules solved in acetonitrile was depleted and the spontaneous Raman scattering at 355 nm suppressed by nearly 50 %. Based on spectroscopic data retrieved from our experiment, we modeled the Raman image of a scattering center in order to demonstrate the applicability of this effect for superresolution Raman microscopy.