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PubMed1979

Gs, an allele of chickens for endogenous avian leukosis viral antigens, segregates with ev 3, a genetic locus that contains structural genes for virus.

Gs is an allele of chickens for the expression of endogenous avian leukosis virus-related core (gs) and envelope (chf) antigens. Progeny of a genetic cross in which Gs was segregating were analyzed for endogenous viral DNA as well as for the expression of endogenous viral antigens. Viral genetic information was identified by cleavage of embryo DNA with restriction endonucleases, electrophoretic separation of the resulting fragments, and identification of bands containing viral sequences by hybridization of the DNA to 32P-labeled viral RNA. Four different chromosomal sites of residence of endogenous viral sequences were identified by this method. These sites were the same as those previously assigned to the endogenous viral loci ev 1, ev 3, ev 4, and ev 5. ev 1 was present in all of the progeny of the cross. ev 3, ev 4, and ev5 were present in various combinations with ev 1. ev 3 cosegregated with the gs+chf+ phenotpye. Cells which did not contain ev 3 but contained ev 1, ev 4, and/or ev 5, did not express detectable levels of viral antigens. We suggest that Gs contains the structural genes for endogenous virus which reside at ev 3 and that these structural genes code for gs and chf in gs+chf+ cells.

Alleles↗
PubMed1980

Ev 2, a genetic locus containing structural genes for endogenous virus, codes for Rous-associated virus type 0 produced by line 72 chickens.

ev 2 is one of seven recently described genetic loci of chickens which contain structural genes for endogenous virus. ev 2 is present exclusively in line 72 chickens, an inbred strain of white Leghorns which is homozygous for the capacity to produce Rous-associated virus type 0 (RAV-0), a subgroup E virus. This phenotype is known as V+ and has been assigned a genetic allele designated V-E7. The segregation of ev 2 was followed in a genetic cross in which the V-E7+ phenotype was also segregating. The progeny of the cross were analyzed for endogenous viral loci by cleavage of embryo DNA with restriction endonuclease SstI, electrophoretic separation of the resulting fragments, and identification of bands containing viral sequences by hybridization of the DNA to radiolabeled viral RNA. Four endogenous viral loci, ev 1, ev 2, ev 4, and ev 5, were identified in the progeny of the cross. One of the progeny contained no detectable endogenous viral sequences. ev 1, ev 4, and ev 5 were present in progeny of both the V-E7+ and V-E7- phenotypes. ev 2 was present exclusively in progeny of the V-E7+ phenotype, and all V-E7+ progeny contained ev 2. In addition, one of the V-E7+ progeny contained only ev 2. FRom these data, we conclude that ev 2 codes for RAV-0 virus produced by the cells of line 72 chickens.

Alleles↗
PubMed2007

Changes of epidermodysplasia verruciformis in benign skin lesions: the EV acanthoma.

Foci of histological changes of epidermodysplasia verruciformis (EV) were noted in five benign skin lesions. These skin lesions included an intradermal nevus, a pigmented seborrheic keratosis, an isolated papule on the forearm, a perianal lesion, and an acantholytic acanthoma. Because the changes resembled true EV so strongly despite the absence of clinical EV in these patients, we searched for EV-human papilloma virus (HPV) types in these skin lesions. Polymerase chain reaction (PCR) analysis on the formalin-fixed, paraffin-embedded blocks was performed. As a positive control, we included tissue from two HIV-positive patients with clinical EV proven by biopsy. Studies were also performed on five other archived biopsies that did not show changes of EV on multiple tissue sections. A nested PCR method detected EV-HPV types in three of the five benign skin lesions showing EV changes as well as in the positive controls. EV changes and EV-HPV can be found incidentally on biopsy in the absence of clinical EV; when such changes are the major histopathological finding in an isolated skin lesion, the lesion should be termed an EV acanthoma.

Acanthoma↗
PubMed1984

[Studies on biological characteristics of enterovirus type 70 (EV 70)].

Enterovirus type 70 (EV 70) is a new type of enterovirus, which is a causative agent of "Acute Hemorrhagic Conjunctivitis" (AHC). It appeared for the first time in Ghana in 1969, and occurred in Japan in 1971. The optimal temperature for growth of EV 70 is 33-35 degrees C, and lower than that of other enteroviruses. Ninety two isolates of EV 70 were investigated to know the biological characteristics. Isolates were obtained in Japan and foreign countries in Africa, Asia and Europe in 1971-1981, and contained eighty seven from eye swabs, three from feces and two from throat swabs. Reproductive capacity of EV 70 at 38 degrees C varied from one isolate to the other. The ratios in log of 33 degrees C yield to 38 degrees C yield ranged from 0.2 to 3.2, and showed no tendency to obtain reproductive capacity at high temperature chronologically after invasion to Japan. In order to find whether EV 70 obtains the reproductive capacity at 38 degrees C, three isolates, which had high pfu ratios (33 degrees C/38 degrees C, log), 2.6-3.2, were subjected to 5 passages by plaquing method. It was found that they obtained the capacities remarkably, and the ratios decreased to -0.5-0.6. As EV 70 exhibits a physicochemical characteristic of enteroviruses, the reproductive capacity in organ cultures of human fetal small intestine was investigated. The results indicated that EV 70 isolates grew distinctly, though the degree of that was less in comparison with coxsackievirus B1 and echovirus 11, and that they increased in the capacities by passages in the intestine. These findings suggested that EV 70 developed the reproductive capacities at high temperature and that in human intestine experimentally. By neutralization tests using two kinds of antisera against EV 70, it was found that serological characteristics of EV 70 changed after invasion to Japan. Intratypic antigenic mutants were first isolated in some parts of Honshu areas in 1972, and took the place of prototype-like isolates throughout Japan in the following one or two years, and after that, no prototype-like isolates obtained anywhere. The same trend was observed in isolates in foreign countries. To investigate differences and changes of these biological characteristics of EV 70 more precisely, further observations seemed to be necessary.

Enterovirus↗
PubMed2026

Human iPSC-EV-loaded nanofiber stent coatings accelerate vascular repair by enhancing EGFR/HIF-1α signaling and suppressing ROCK1-mediated remodeling.

Arterial disease management is shifting from antiproliferative drug-eluting stents toward approaches that restore endothelial function and modulate smooth muscle cell (SMC) behavior. Stem cell-derived extracellular vesicles (EVs) carry miRNAs that promote endothelial proliferation and migration while restraining aberrant SMC growth and inflammation. Here, human induced pluripotent stem cell (iPSC)-derived EVs were collected by ultracentrifugation and incorporated into 50:50 poly (lactic-co-glycolic acid) (PLGA 503) core-shell nanofibrous membranes, which were fabricated as stent coatings for sustained release to overcome rapid clearance and poor tissue retention. EVs derived from three independent iPSC lines all enhanced tube formation in human umbilical vein endothelial cells (HUVECs) under hypoxic and serum-starved conditions and revealed a trend toward reduced platelet-derived growth factor-BB (PDGF-BB)-induced smooth muscle cell (SMC) migration. The fabricated core-shell nanofibers enabled sustained EV release, maintaining therapeutic efficacy for 28 days. Small RNA sequencing (NGS) analysis demonstrated that EVs from these independent iPSC lines shared miR-148a-3p and members of the miR-92 family, which collectively accounted for more than 75% of the reads within the 25 top-expressed miRNA set. In vitro, iPSC-EVs enhanced HUVEC proliferation and survival signaling by downregulating the negative regulators ERRFI1 and VHL, which are specific targets of miR-148a-3p and the miR-92 family, thereby activating the EGFR and HIF-1α axes and driving downstream ERK1/2 and VEGF expression under hypoxic and serum starvation stress conditions. Concurrently, iPSC-EVs prevented PDGF-BB-induced SMC phenotypic switching by downregulating ROCK1, a target of miR-148a-3p, thereby inhibiting downstream AKT and ERK signaling and preserving contractile markers while suppressing the synthetic phenotype. In vivo, the iPSC-EV-functionalized scaffolds significantly accelerated re-endothelialization and inhibited neointimal hyperplasia, evidenced by the upregulation of angiogenic factors (VEGF, CD31) and the concurrent suppression of pathological remodeling markers (α-SMA, MMPs) and inflammatory cytokines (IL-6, TGF-β1). Therefore, iPSC-EVs enriched with specific miRNAs and delivered via PLGA 503 core-shell nanofibers promote endothelial repair while suppressing SMC overgrowth, providing a promising strategy for vascular healing.

Core-shell nanofibers↗
PubMed2007

Electron photoemission from charged films: absolute cross section for trapping 0-5 eV electrons in condensed CO2.

The electron trapping or attachment cross section of carbon dioxide (CO2) condensed as thin films on a spacer of Ar is obtained using a simple model for electron trapping in a molecular film and then charge releasing from the same film by photon absorption. The measurements are presented for different electron exposures and impact energies, film thicknesses, and probing photon energies. The cross section for trapping an electron of incident energy between 0 and 5 eV reveals three different attachment processes characterized by a maximum at about 0.75 eV, a structured feature around 2.25 eV, and a shoulder around 3.75 eV. From the measurement of their dependence with the probing photon energy, the two lowest processes produce traps having a vertical electron binding energy of approximately 3.5 eV, whereas the highest one yields a slightly higher value of approximately 3.7 eV. The 0.75 eV maximum corresponds to the formation of vibrational Feshbach resonances in (CO2)n- anion clusters. The 2.25 eV feature is attributed to the formation of a vibrationally excited 2Piu anion in (CO2)n- clusters, followed by fast decay into its vibrational ground state without undergoing autodetachment. Finally, 3.75 eV shoulder is assigned to the well-known dissociative electron attachment process from 2Piu anion state producing the O- anion in the gas phase and the (CO2)nO- anions in clusters.

Carbon Dioxide↗
PubMed2005

Absolute vibrational and electronic cross sections for low-energy electron (2-12 eV) scattering from condensed pyrimidine.

Low-energy vibrational and electronic electron-energy-loss (EEL) spectra of pyrimidine condensed on a thin film of solid argon held at 18 K are reported for the incident-energy range of 2-12 eV. Sensitivity to symmetry and spin forbidden transitions as well as correlations to the triplet states of benzene make it possible to ascribe the main features, below 7 eV in the electronic part of the EEL spectrum, to triplet transitions. The lowest EEL feature with an energy onset at 3.5 eV is attributed to a transition to the (3)B(1)(n-->pi(*)) valence electronic state and the next triplet n-->pi(*) transition to a (3)A(2) state located around 4.5 eV. The remaining EEL features at 4.3, 5.2, 5.8, and 6.5 eV are all assigned to pi-->pi(*) transitions to states of symmetry (3)B(2), (3)A(1), (3)B(2), and (3)B(2)+(3)A(1), respectively. The most intense maximum at 7.6 eV is found to correspond to both (1)B(2) and (1)A(1) transitions, as in the vacuum ultraviolet spectra. Absolute inelastic cross sections per scatterer are derived from a single collision treatment described herein. Their values are found to lie within the 10(-17) cm(2) range for both the electronic and the vibrational excitations. Features in the energy dependence of the cross sections are discussed, whenever possible, by comparison with data and mechanisms found in the gas phase. A maximum over the 4-5 eV range is attributed to a B (2)B(1) shape resonance and another one observed in the 6-7 eV range is ascribed to either or both sigma(*) shape resonances of (2)A(1) and (2)B(2) symmetries.

Journal Article↗
PubMed1982

Role of methylation in the induced and spontaneous expression of the avian endogenous virus ev-1: DNA structure and gene products.

The endogenous avian provirus ev-1 is widespread in white leghorn chickens. Although it has no major structural defects, ev-1 has not been associated with any phenotype and is ordinarily expressed at a very low level. In this report, we describe a chicken embryo (Number 1836) cell culture containing both ev-1 and ev-6 which spontaneously expressed the ev-1 provirus. This culture released a high level of noninfectious virions containing a full complement of virion structural (gag) proteins but devoid of reverse transcriptase activity or antigen. These virions contained 70S RNA closely related to the genome of Rous-associated virus type 0, but identifiable as the ev-1 genome by oligonucleotide mapping. A fraction of the RNA molecules in the 70S complex were unusual in that they were polyadenylated 100 to 200 nucleotides downstream of the usual polyadenylation site. Eight sibling embryo cultures did not share this unusual phenotype with 1836, indicating that it was not inherited. However, an identical phenotype was inducible in the sibling cultures by treatment with 5-azacytidine, an inhibitor of DNA methylation, and the induced expression was stable for more than 10 generations. Analysis of chromatin structure and DNA methylation of the ev-1 provirus in 1836 cells revealed the presence (in a fraction of the proviruses) of both DNase I hypersensitive sites in the long terminal repeats and in gag and a pattern of cleavage sites for methyl-sensitive restriction endonuclease not found in a nonexpressing sibling. These results lend strong support to the role of DNA methylation in the control of gene expression. Additionally, they explain the lack of phenotype associated with ev-1 as due to a combination of its low expression and defectiveness in pol and env.

Animals↗
PubMed2025

EV-D68 cleaves LARP1 and PABPC1 by 3Cpro to redirect host mRNA translation machinery toward its genomic RNA.

Enterovirus D68 (EV-D68) is an emerging pathogen associated with severe respiratory diseases and neurological complications, such as acute flaccid myelitis. EV-D68 has developed sophisticated mechanisms to hijack host translation machinery, facilitating its replication and impairing host mRNA translation. In this study, we demonstrate that EV-D68 cleaves La-related protein 1 (LARP1) and poly(A)-binding protein cytoplasmic 1 (PABPC1) through its proteases 3Cpro and 2Apro. Our results indicate that overexpressing LARP1 and PABPC1 significantly inhibits EV-D68 replication and reduces the virus-mediated suppression of host translation. While both LARP1 and PABPC1 regulate translation, they exert antiviral effects through distinct mechanisms. We found that LARP1 interacts with the 5'UTR of EV-D68 RNA through its LAM domain, and this interaction is crucial for its antiviral function. LARP1 translation modulation is also influenced by the mTOR and CDK1 signaling pathways. Viral infection inhibits mTOR and CDK1 phosphorylation, which enhances LARP1's binding to viral RNA and inhibits viral translation. To counteract this inhibition, EV-D68 cleaves LARP1 through 3Cpro, thereby promoting efficient viral translation. We also investigated other enteroviruses, such as EV-A71 and CV-A16, which similarly target LARP1 and PABPC1, indicating a conserved mechanism across enteroviruses. Our findings offer new insights into how EV-D68 manipulates host translation and highlight the potential of targeting LARP1 and PABPC1 for antiviral interventions.

Humans↗
PubMed2004

HPV typing in Brazilian patients with epidermodysplasia verruciformis: high prevalence of EV-HPV 25.

BACKGROUND: Epidermodysplasia verruciformis is a rare genetic disorder characterized by development of lesions associated with HPV#5 or HPV#8 in early childhood; malignant transformation occurs in approximately half of individuals during adulthood. OBJECTIVE: Our goal was to study the presence and spectrum of EV-HPV types in Brazilian EV patients, a population that had never been studied in this regard. PATIENTS AND METHODS: Forty-one biopsies from different lesions (benign and skin tumors) and one biopsy from clinically normal skin from each of 20 Brazilian patients with EV were studied for HPV typing using nested PCR. RESULTS: EV-HPV DNA was detected in all 41 skin lesions of the patients and was also identified in specimens considered as normal skin from 8 patients (40%). In this study HPV-EV 25 was the most prevalent (70%), and HPV 14d (67%) was highly associated with malignant lesions. CONCLUSION: EV-HPV 25 was the most prevalent in our study. The noteworthy association of EV-HPV type 14d with skin cancers suggests its possible oncogenic role in malignant transformation in this population.

Adolescent↗
PubMed2003

Single, double, and multiple double strand breaks induced in DNA by 3-100 eV electrons.

Nonthermal secondary electrons with initial kinetic energies below 100 eV are an abundant transient species created in irradiated cells and thermalize within picoseconds through successive multiple energy loss events. Here we show that below 15 eV such low-energy electrons induce single (SSB) and double (DSB) strand breaks in plasmid DNA exclusively via formation and decay of molecular resonances involving DNA components (base, sugar, hydration water, etc.). Furthermore, the strand break quantum yields (per incident electron) due to resonances occur with intensities similar to those that appear between 25 and 100 eV electron energy, where nonresonant mechanisms related to excitation/ionizations/dissociations are shown to dominate the yields, although with some contribution from multiple scattering electron energy loss events. We also present the first measurements of the electron energy dependence of multiple double strand breaks (MDSB) induced in DNA by electrons with energies below 100 eV. Unlike the SSB and DSB yields, which remain relatively constant above 25 eV, the MDSB yields show a strong monotonic increase above 30 eV, however with intensities at least 1 order of magnitude smaller than the combined SSB and DSB yields. The observation of MDSB above 30 eV is attributed to strand break clusters (nano-tracks) involving multiple successive interactions of one single electron at sites that are distant in primary sequence along the DNA double strand, but are in close contact; such regions exist in supercoiled DNA (as well as cellular DNA) where the double helix crosses itself or is in close proximity to another part of the same DNA molecule.

DNA↗
PubMed2005

Absolute electronic excitation cross sections for low-energy electron (5-12 eV) scattering from condensed thymine.

The absolute cross sections for electronic excitations of thymine by electron impact between 5 and 12 eV are determined by means of electron-energy loss (EEL) spectroscopy for the molecule deposited at submonolayer coverage on an inert Ar substrate. The lowest EEL features at 3.7 and 4.0 eV are attributed to the excitation of the triplet 1 3A'(pi --> pi*) and 1 3A''(n --> pi*) valence states of the molecule. The higher EEL features located at 4.9, 6.3, 7.3, and 9 eV with a weak shoulder around 6 eV are ascribed mostly to triplet valence (pi --> pi*) excitation manifold of the molecule. The energy dependence of the cross section for both the lowest triplet valence excitations shows essentially a peak at about 5 eV reaching a value of 2.9 x 10(-17) cm2. The cross sections for the higher EEL features are generally characterized by a common broad maximum around 8 eV. The latter reaches a value of 1.36 x 10(-16) cm2 for the combined 6 and 6.3 eV excitation region. The maxima in the present cross sections are found to correspond to the resonances that have been reported at about the same energies in the O- yield from electron impact on thymine in the gas phase.

Models, Chemical↗
PubMed1983

The influence of the ev 3 locus on the inducibility of serum antibody reactivity for envelope glycoprotein group-specific determinants.

Chickens segregating for the ev 3 locus were bred by backcross matings of line 6(3) to line 15B. Analysis of RAV-1-infected segregants indicated that inducibility of antibody reactivity for envelope glycoprotein group-specific determinants correlated with the absence of ev 3, whereas noninducibility correlated with the presence of ev 3. Since the ev 3(+) and ev 3(-) segregants possessed similar genetic backgrounds, these results provide direct evidence that the ev 3 locus determines the phenotype of noninducibility.

Animals↗
PubMed1984

Individual ev loci determine characteristic patterns of endogenous retroviral envelope antigen expression during lymphocyte maturation.

A comparative study of the expression of retroviral envelope antigen by T and B lymphocytes at different maturational stages was made with chickens typed for genomic ev loci. All lymphocyte subclasses from chf(-) chickens were negative for expression of retroviral envelope antigen. By contrast, cells of the lymphocyte lineage from all the chf(+) lines examined were positive for expression, with the levels of envelope antigen in the lymphoblasts exceeding the levels in the immature and resting cells. This pattern of lymphocyte expression was found in lines of chickens in which the expression of chf in fibroblasts was encoded by the ev 3, ev 6, or ev 9 loci. Plasma cells from those chf(+) lines possessing ev 6 expressed higher levels of envelope antigen than the lymphoblasts, an effect not observed with the chf(+) lines lacking ev 6.

Animals↗
PubMed2025

Streptococcus pyogenes EVs induce the alternative inflammasome via caspase-4/-5 in human monocytes.

The sensing of Gram-negative Extracellular Vesicles (EVs) by the innate immune system has been extensively studied in the past decade. In contrast, recognition of Gram-positive EVs by innate immune cells remains poorly understood. Comparative genome-wide transcriptional analysis in human monocytes uncovered that S. pyogenes EVs induce proinflammatory signatures that are markedly distinct from those of their parental cells. Among the 209 genes exclusively upregulated by EVs, caspase-5 prompted us to study inflammasome signaling pathways in depth. We show that lipoteichoic acid (LTA), a structural component of Gram-positive bacterial membranes present on EVs from S. pyogenes and other Gram-positive species, is sensed by TLR2 which triggers the alternative inflammasome composed of NLRP3 and the inflammatory caspases-4/-5 to mount an IL-1β response without inducing cell death. For S. pyogenes, we identify TLR8 as a sensor to mediate caspase-4/-5-dependent IL-1β secretion. Notably, inflammasome activation by intact bacteria is independent of the global virulence regulator CovS in monocytes. Overall, our study highlights a new role for TLR2 and caspase-4/-5 in the recognition of Gram-positive EVs in human monocytes.

Humans↗
PubMed2002

Fragmentation of short single DNA strands by 1-30 eV electrons: dependence on base identity and sequence.

PURPOSE: To investigate the dependence of base identity and sequence on the damage induced by low-energy (1-30 eV) electron impact on a short single strand of DNA. MATERIALS AND METHODS: Monolayers of homogeneous nonamers of deoxycytidine and thymidine (dCy9 and T9) and heterogeneous nonamers of thymidine substituted with 33 and 66% of deoxycytidine (dCy3-T6 and dCy6-T3) were chemisorbed onto a gold substrate. They were bombarded under ultrahigh vacuum conditions by a 1-30 eV electron beam. Neutral fragments desorbed from the films were detected by a mass spectrometer. From partial pressure measurements, the effective cross-section (ECS) per base for desorption of various fragments was estimated. RESULTS: CN, OCN and/or H2NCN were the major neutral species observed to desorb in the present experiments. A small contribution of 55 amu neutral species, tentatively attributed to CH3CCO, were only detected from fragmentation of oligonucleotides containing thymine. The total ECS per base estimated for the CN, OCN and CH3CCO species production from fragmentation of dCy9, dCy6-T3, dCy3-T6 and T9 at 12 eV incident electron energy were (3.4, 2.0, 2.9 and 2.3) x 10(-17) cm(2), respectively. The incident electron energy dependence of ECS for desorption of these fragments exhibited structures <20 eV, which are characteristic of transient anion formation. CONCLUSIONS: At incident electron energies <20 eV, neutral fragment desorption arise from dissociation of the DNA bases, principally via dissociative electron attachment and/or decay of the transient anion into a dissociative electronic excited state of the base. Non-resonant mechanisms (e.g. direct dipolar dissociation) mostly control the fragmentation processes >20 eV. From comparison of the electron energy dependence of the ECS for base fragmentation in the homo- and heteronucleotides, it is concluded that damage to a short DNA strand is dependent on base identity, sequence and electron energy.

Anions↗
PubMed1994

Single- and double-strand breaks in pBR322 DNA by vacuum-UV from 8.3 to 20.7 eV.

Solid pBR322 DNA was irradiated in a vacuum by monochromatic photons from 8.3 eV (150 nm) to 20.7 eV (60 nm), and the formation of single- and double-strand breaks (ssb and dsb) was determined using agarose gel electrophoresis. The action cross sections increased 20 times (ssb) and 43 times (dsb) from 8.3 eV to 20.7 eV; the quantum yields increased 5.4 times (ssb) and 12 times (dsb). The cross sections for dsb were 0.0059 at 8.3 eV and 0.013 at 20.7 eV of those of ssb; these values were smaller than about 0.04 for 2.1 keV photons, or about 0.18 for 60Co gamma-rays. Although vacuum UV photons were proved to induce dsb, they had a lower efficiency than when using soft X-rays or gamma-rays.

DNA↗
PubMed1993

Genetic analysis of four differing traits (MHC haplotype, tumor response, CD4:CD8 ratio, ev 17) in CB and CC congenic chickens.

Two closely related inbred lines of chickens, CB and CC, were compared for major histocompatibility complex (MHC) haplotype, response to tumors induced by Rous sarcoma virus and CD4:CD8 ratio. Genetic analysis showed that all the above traits were associated with the MHC. In all CB chickens, we found an additional endogenous virus, ev 17, a proviral sequence lacking in all tested CC animals. From our analysis and data from the literature, we presume that the presence of ev 17 in CB chickens is probably caused by gene duplication of ev 1 with extensive nucleotide alterations. Our analysis with (CB x CC) x CC backcrosses demonstrate that ev 17 is not linked to the chicken B complex. From our experiments, we conclude that inbred CB and CC chickens differ in MHC and ev 17, and we established additional congenic inbred lines with and without the proviral locus ev 17.

Animals↗