Expression of Epstein–Barr Virus–Encoded Small RNA (by the EBER-1 Gene) in Liver Specimens from Transplant Recipients with Post-Transplantation Lymphoproliferative Disease

Epstein-Barr virus (EBV)—associated post-transplantation lymphoproliferative disease (PTLD) develops in 1 to 10 percent of transplant recipients, in whom it can be treated by a reduction in the level of immunosuppression. We postulated that the tissue expression of the small RNA transcribed by the EBER-1 gene during latent EBV infection would identify patients at risk for PTLD. We studied EBER-1 gene expression in liver specimens obtained from 24 patients 2 days to 22 months before the development of PTLD, using in situ hybridization with an oligonucleotide probe. Control specimens were obtained from 20 recipients of allografts with signs of injury due to organ retrieval, acute graft rejection, or viral hepatitis in whom PTLD had not developed 9 to 71 months after the biopsy. Of the 24 patients with PTLD, 17 (71 percent) had specimens in which 1 to 40 percent of mononuclear cells were positive for the EBER-1 gene. In addition, 10 of these 17 patients (59 percent) had specimens with histopathological changes suggestive of EBV hepatitis. In every case, EBER-1—positive cells were found within the lymphoproliferative lesions identified at autopsy. Only 2 of the 20 controls (10 percent) had specimens with EBER-1—positive cells (P<0.001), and such cells were rare. EBER-1 gene expression in liver tissue precedes the occurrence of clinical and histologic PTLD. The possibility of identifying patients at risk by the method we describe here and preventing the occurrence of PTLD by a timely reduction of immunosuppression needs to be addressed by future prospective studies. (N Engl J Med 1992;327:1710–4.), POST-TRANSPLANTATION lymphoproliferative disease (PTLD), either polyclonal or monoclonal, complicates the clinical course of 1 to 10 percent of organ-transplant recipients.123 Immunohistochemical studies have demonstrated that the lymphoid cells within the lesions of PTLD almost invariably contain Epstein–Barr virus (EBV), primarily in a state of latent infection.4,5 The EBER-1 gene is expressed early during latent EBV infection and codes for a small messenger RNA (mRNA) expressed at up to 107 copies per cell.6 We and others have previously demonstrated the value of the detection of EBER-1 RNA for identifying EBV-infected cells in formalin-fixed paraffin-embedded tissues.7,8 In the current investigation, we used… © 1992, Massachusetts Medical Society. All rights reserved

The interferon-induced exonuclease ISG20 exerts antiviral activity through upregulation of type I interferon response proteins

The host immune responses to infection lead to the production of type I interferon (IFN), and the upregulation of interferon-stimulated genes (ISGs) reduces virus replication and virus dissemination within a host. Ectopic expression of the interferon-induced 20-kDa exonuclease ISG20 suppressed replication of chikungunya virus and Venezuelan equine encephalitis virus, two mosquito-vectored RNA alphaviruses. Since the replication of alphavirus genomes occurs exclusively in the cytoplasm, the mechanism of nucleus-localized ISG20 inhibition of replication is unclear. In this study, we determined that ISG20 acts as a master regulator of over 100 genes, many of which are ISGs. Specifically, ISG20 upregulated IFIT1 genes and inhibited translation of the alphavirus genome. Furthermore, IFIT1-sensitive alphavirus replication was increased in Isg20−/− mice compared to the replication of wild-type viruses but not in cells ectopically expressing ISG20. We propose that ISG20 acts as an indirect regulator of RNA virus replication in the cytoplasm through the upregulation of many other ISGs.Type I interferon (IFN)-stimulated genes (ISGs) have critical roles in inhibiting virus replication and dissemination. Despite advances in understanding the molecular basis of ISG restriction, the antiviral mechanisms of many remain unclear. The 20-kDa ISG ISG20 is a nuclear 3′–5′ exonuclease with preference for single-stranded RNA (ssRNA) and has been implicated in the IFN-mediated restriction of several RNA viruses. Although the exonuclease activity of ISG20 has been shown to degrade viral RNA in vitro, evidence has yet to be presented that virus inhibition in cells requires this activity. Here, we utilized a combination of an inducible, ectopic expression system and newly generated Isg20−/− mice to investigate mechanisms and consequences of ISG20-mediated restriction. Ectopically expressed ISG20 localized primarily to Cajal bodies in the nucleus and restricted replication of chikungunya and Venezuelan equine encephalitis viruses. Although restriction by ISG20 was associated with inhibition of translation of infecting genomic RNA, degradation of viral RNAs was not observed. Instead, translation inhibition of viral RNA was associated with ISG20-induced upregulation of over 100 other genes, many of which encode known antiviral effectors. ISG20 modulated the production of IFIT1, an ISG that suppresses translation of alphavirus RNAs. Consistent with this observation, the pathogenicity of IFIT1-sensitive alphaviruses was increased in Isg20−/− mice compared to that of wild-type viruses but not in cells ectopically expressing ISG20. Our findings establish an indirect role for ISG20 in the early restriction of RNA virus replication by regulating expression of other ISGs that inhibit translation and possibly other activities in the replication cycle

Validating Spray Coverage Rate Using Liquid Mass on a Spray Card

Validation of agricultural sprayers is important for quantifying as-applied coverage rates under field conditions. The complexity of modern sprayer control systems presents a challenge for precise field validation due to the use of nozzle control technologies, such as pulse width modulation, to meter chemical flow rates at individual nozzles. Non-uniform flow over time may result in local variations at high spatial resolutions that are ignored when estimating as-applied coverage rates across a field. The purpose of this study was to test several methods for estimating the mass of water applied to a water-sensitive paper spray card target using steady-state and instantaneous measurement techniques. The steady-state method consisted of a spray patternator table used to quantify the mass flow rate distribution across the nozzle width at varying nozzle pressures. The mass flow rate was then projected onto a two-dimensional area traveling across the spray width to calculate the mass of water that was deposited in the area. Two instantaneous sampling methods were used. The first method directly measured the mass of the spray card and water for 5 min after exposure to model the evaporation rate and solve for the initial mass at the time of exposure. The second method indirectly used the percent coverage of the exposed spray card by droplets. Results showed that the error between the calculated mass of water from the mass flow rate and the estimated initial mass of water from the evaporation rate varied between 2% and 8%. The relationships between the calculated and estimated initial mass of water methods and the spray card percent coverage were highly linear (R2 \u3e 0.98). Both instantaneous methods produced results with higher variability between replications than the steady-state method, but the number of replications resulted in acceptably small differences between average mass measurements. These results show the potential for using evaporation rates for laboratory validation and percent coverage for laboratory or field validation of as-applied coverage rates

Rigorous Born Approximation and beyond for the Spin-Boson Model

Within the lowest-order Born approximation, we present an exact calculation of the time dynamics of the spin-boson model in the ohmic regime. We observe non-Markovian effects at zero temperature that scale with the system-bath coupling strength and cause qualitative changes in the evolution of coherence at intermediate times of order of the oscillation period. These changes could significantly affect the performance of these systems as qubits. In the biased case, we find a prompt loss of coherence at these intermediate times, whose decay rate is set by $\sqrt{\alpha}$, where $\alpha$ is the coupling strength to the environment. We also explore the calculation of the next order Born approximation: we show that, at the expense of very large computational complexity, interesting physical quantities can be rigorously computed at fourth order using computer algebra, presented completely in an accompanying Mathematica file. We compute the $O(\alpha)$ corrections to the long time behavior of the system density matrix; the result is identical to the reduced density matrix of the equilibrium state to the same order in $\alpha$. All these calculations indicate precision experimental tests that could confirm or refute the validity of the spin-boson model in a variety of systems.Comment: Greatly extended version of short paper cond-mat/0304118. Accompanying Mathematica notebook fop5.nb, available in Source, is an essential part of this work; it gives full details of the fourth-order Born calculation summarized in the text. fop5.nb is prepared in arXiv style (available from Wolfram Research

User frustrations as opportunities

User frustrations are an excellent source of new product ideas. Starting with this observation, this article describes an approach that entrepreneurs can use to discover business opportunities. Opportunity discovery starts with a problem that the user has, but may not be able to articulate. User-centered design techniques can help elicit those latent needs. The entrepreneur should then try to understand how users are solving their problem today, before proposing a solution that draws on the unique skills and technical capabilities available to the entrepreneur. Finally, an in-depth understanding of the user allows the entrepreneur to hone in on the points of difference and resonance that are the foundation of a strong customer value proposition