Pleural effusions are common, with an estimated 1-1.5 million new cases in the United States and 200 000-250 000 in the United Kingdom each year. Rahul Bhatnagar, academic clinical lecturer at the University of Bristol, describes how pleural effusions may be investigated and treated in the community and secondary care, with a particular focus on...

Insulin-mediated microvascular recruitment (IMVR) regulates delivery of insulin and glucose to insulin-sensitive tissues. We have previously proposed that perivascular adipose tissue (PVAT) controls vascular function through outside-to-inside communication and through vessel-to-vessel, or "vasocrine," signaling. However, direct experimental evidence supporting a role of local PVAT in regulating IMVR and insulin sensitivity in vivo is lacking. Here, we studied muscles with and without PVAT in mice using combined contrast-enhanced ultrasonography and intravital microscopy to measure IMVR and gracilis artery diameter at baseline and during the hyperinsulinemic-euglycemic clamp. We show, using microsurgical removal of PVAT from the muscle microcirculation, that local PVAT depots regulate insulin-stimulated muscle perfusion and glucose uptake in vivo. We discovered direct microvascular connections between PVAT and the distal muscle microcirculation, or adipomuscular arterioles, the removal of which abolished IMVR. Local removal of intramuscular PVAT altered protein clusters in the connected muscle, including upregulation of a cluster featuring Hsp90ab1 and Hsp70 and downregulation of a cluster of mitochondrial protein components of complexes III, IV, and V. These data highlight the importance of PVAT in vascular and metabolic physiology and are likely relevant for obesity and diabetes.

Optimal immune-based therapies for type 1 diabetes (T1D) should restore self-tolerance without inducing chronic immunosuppression. CD4⁺Foxp3⁺ regulatory T cells (T_regs) are a key cell population capable of facilitating durable immune tolerance. However, clinical trials with expanded T_regs in T1D and solid-organ transplant recipients are limited by poor T_reg engraftment without host manipulation. We showed that T_reg engraftment and therapeutic benefit in nonautoimmune models required ablative host conditioning. Here, we evaluated T_reg engraftment and therapeutic efficacy in the nonobese diabetic (NOD) mouse model of autoimmune diabetes using nonablative, combinatorial regimens involving the anti-CD3 (αCD3), cyclophosphamide (CyP), and IAC (IL-2/JES6–1) antibody complex. We demonstrate that αCD3 alone induced substantial T-cell depletion, impacting both conventional T cells (T_conv) and T_regs, subsequently followed by more rapid rebound of T_regs. Despite robust depletion of host T_conv and host T_regs, donor T_regs failed to engraft even with interleukin-2 (IL-2) support. A single dose of CyP after αCD3 depleted rebounding host T_regs and resulted in a 43-fold increase in donor T_reg engraftment, yet polyclonal donor T_regs failed to reverse diabetes. However, infusion of autoantigen-specific T_regs after αCD3 alone resulted in robust T_reg engraftment within the islets and induced remission in all mice. This novel combinatorial therapy promotes engraftment of autoantigen-specific donor T_regs and controls islet autoimmunity without long-term immunosuppression.

OBJECTIVE

To examine differences in myocardial blood flow (MBF) at rest and during stress between patients with type 2 diabetes and controls, and to identify potential predictors of changes in MBF at rest and during stress.

OBJECTIVE

We conducted an open-label, phase I study using autologous umbilical cord blood (UCB) infusion to ameliorate type 1 diabetes (T1D). Having previously reported on the first 15 patients reaching 1 year of follow-up, herein we report on the complete cohort after 2 years of follow-up.

[London] : [Thomas. Bowles] : [John Bowles], [1728]

Gregor Pasemann, Wilhelm Stannat.

Source: Electronic Journal of Statistics, Volume 14, Number 1, 547--579.

Abstract:
A parameter estimation problem for a class of semilinear stochastic evolution equations is considered. Conditions for consistency and asymptotic normality are given in terms of growth and continuity properties of the nonlinear part. Emphasis is put on the case of stochastic reaction-diffusion systems. Robustness results for statistical inference under model uncertainty are provided.

We propose a new semiparametric approach for modelling nonlinear univariate diffusions, where the observed process is a nonparametric transformation of an underlying parametric diffusion (UPD). This modelling strategy yields a general class of semiparametric Markov diffusion models with parametric dynamic copulas and nonparametric marginal distributions. We provide primitive conditions for the identification of the UPD parameters together with the unknown transformations from discrete samples. Likelihood-based estimators of both parametric and nonparametric components are developed and we analyze the asymptotic properties of these. Kernel-based drift and diffusion estimators are also proposed and shown to be normally distributed in large samples. A simulation study investigates the finite sample performance of our estimators in the context of modelling US short-term interest rates. We also present a simple application of the proposed method for modelling the CBOE volatility index data.

Uncertainties in a structure is inevitable, which generally lead to variation in dynamic response predictions. For a complex structure, brute force Monte Carlo simulation for response variation analysis is infeasible since one single run may already be computationally costly. Data driven meta-modeling approaches have thus been explored to facilitate efficient emulation and statistical inference. The performance of a meta-model hinges upon both the quality and quantity of training dataset. In actual practice, however, high-fidelity data acquired from high-dimensional finite element simulation or experiment are generally scarce, which poses significant challenge to meta-model establishment. In this research, we take advantage of the multi-level response prediction opportunity in structural dynamic analysis, i.e., acquiring rapidly a large amount of low-fidelity data from reduced-order modeling, and acquiring accurately a small amount of high-fidelity data from full-scale finite element analysis. Specifically, we formulate a composite neural network fusion approach that can fully utilize the multi-level, heterogeneous datasets obtained. It implicitly identifies the correlation of the low- and high-fidelity datasets, which yields improved accuracy when compared with the state-of-the-art. Comprehensive investigations using frequency response variation characterization as case example are carried out to demonstrate the performance.

Anton Thalmaier, James Thompson.

Source: Bernoulli, Volume 26, Number 3, 2202--2225.

Abstract:
In this article, we derive moment estimates, exponential integrability, concentration inequalities and exit times estimates for canonical diffusions firstly on sub-Riemannian limits of Riemannian foliations and secondly in the nonsmooth setting of $operatorname{RCD}^{*}(K,N)$ spaces. In each case, the necessary ingredients are Itô’s formula and a comparison theorem for the Laplacian, for which we refer to the recent literature. As an application, we derive pointwise Carmona-type estimates on eigenfunctions of Schrödinger operators.

Arnak S. Dalalyan, Lionel Riou-Durand.

Source: Bernoulli, Volume 26, Number 3, 1956--1988.

Abstract:
Langevin diffusion processes and their discretizations are often used for sampling from a target density. The most convenient framework for assessing the quality of such a sampling scheme corresponds to smooth and strongly log-concave densities defined on $mathbb{R}^{p}$. The present work focuses on this framework and studies the behavior of the Monte Carlo algorithm based on discretizations of the kinetic Langevin diffusion. We first prove the geometric mixing property of the kinetic Langevin diffusion with a mixing rate that is optimal in terms of its dependence on the condition number. We then use this result for obtaining improved guarantees of sampling using the kinetic Langevin Monte Carlo method, when the quality of sampling is measured by the Wasserstein distance. We also consider the situation where the Hessian of the log-density of the target distribution is Lipschitz-continuous. In this case, we introduce a new discretization of the kinetic Langevin diffusion and prove that this leads to a substantial improvement of the upper bound on the sampling error measured in Wasserstein distance.

Daniela Pauger, Helga Wagner.

Source: Bayesian Analysis, Volume 14, Number 2, 341--369.

Abstract:
We propose a Bayesian approach to obtain a sparse representation of the effect of a categorical predictor in regression type models. As this effect is captured by a group of level effects, sparsity cannot only be achieved by excluding single irrelevant level effects or the whole group of effects associated to this predictor but also by fusing levels which have essentially the same effect on the response. To achieve this goal, we propose a prior which allows for almost perfect as well as almost zero dependence between level effects a priori. This prior can alternatively be obtained by specifying spike and slab prior distributions on all effect differences associated to this categorical predictor. We show how restricted fusion can be implemented and develop an efficient MCMC (Markov chain Monte Carlo) method for posterior computation. The performance of the proposed method is investigated on simulated data and we illustrate its application on real data from EU-SILC (European Union Statistics on Income and Living Conditions).

Georg S. Kranz
Nov 12, 2014; 34:15466-15475
Systems/Circuits

Italian translation of Press Release about BIS General Manager Agustín Carstens giving a speech on "Money in the digital age: what role for central banks?" (6 February 2018)

The Saskatchewan Union of Nurses (SUN) is raising concerns about some nurses who volunteered for remote assignments being told they need to refrain from working elsewhere for at least 14 days if they’ve worked in Lloydminster or La Loche.

It will be a difficult road back to any kind of normal living.

OM Ukraine's sports team equips and trains churches in using non-traditional sports in their summer camps.

Doxorubicin therapy, effective against many malignancies, is limited by cardiotoxicity. Continuous-infusion doxorubicin, compared with bolus-infusion, reduces early cardiotoxicity in adults. Its effectiveness in reducing late cardiotoxicity in children remains uncertain.

This multicenter randomized trial assessed whether continuous-infusion of doxorubicin in pediatric patients provides long-term cardioprotection or improvement in event-free survival over bolus-infusion in acute lymphoblastic leukemia. Continuous-infusion of doxorubicin provided no cardioprotection or improvement in event-free survival. (Read the full article)

High bilirubin levels are associated with sensorineural hearing loss. Exchange transfusions are recommended when bilirubin levels reach certain thresholds. However, the relative and excess risks of hearing loss in infants with bilirubin levels at/above exchange transfusion thresholds are unknown.

In this Northern California population of term and late preterm infants, elevated bilirubin levels were not associated with an increased risk of sensorineural hearing loss unless the levels were at least 10 mg/dL above exchange transfusion thresholds. (Read the full article)

Pharmacokinetic changes are often seen in patients with severe infections. Administration by continuous infusion has been suggested to optimize antibiotic exposure and pharmacokinetic/pharmacodynamic (PK/PD) target attainment for β-lactams. In an observational study, unbound piperacillin concentrations (n=196) were assessed in 78 critically ill patients following continuous infusion of piperacillin/tazobactam (ratio 8:1). The initial dose of 8, 12 or 16 g (piperacillin component) was determined by individual creatinine clearance (CRCL). Piperacillin concentrations were compared to the EUCAST clinical breakpoint MIC for Pseudomonas aeruginosa (16 mg/L), and the following PK/PD targets were evaluated: 100% fT>1xMIC and 100% fT>4xMIC. A population pharmacokinetic model was developed using NONMEM 7.4.3 consisting of a one-compartment disposition model with linear elimination separated into non-renal and renal (linearly increasing with patient CRCL) clearances. Target attainment was predicted and visualized for all individuals based on the utilized CRCL dosing algorithm. The target of 100% fT>1xMIC was achieved for all patients based on the administered dose, but few patients achieved the target of 100% fT>4xMIC. Probability of target attainment for a simulated cohort of patients showed, that increasing the daily dose by 4 g increments (piperacillin component) did not result in substantially improved target attainment for the 100% fT>4xMIC target. To conclude, in patients with high CRCL combined with high-MIC bacterial infections, even a CI regimen with a daily dose of 24 g may be insufficient to achieve therapeutic concentrations.

BACKGROUND AND OBJECTIVES:

Annual incidence of venous thromboembolism (VTE) including postoperative VTE in hospitalized children is rising significantly. A growing body of evidence supports the role of red blood cells (RBCs) in pathologic thrombosis. In this study, we examined the association of perioperative RBC transfusion with postoperative VTE in pediatric patients.

Infusionsoft by Keap combines robust analytics with an intuitive interface and easily customizable dashboards to help you track your customers and email marketing campaigns.

The Education Department says federal law should not be used to prevent schools from offering online learning to all students, including those with disabilities.

IT’S a bank holiday weekend and we are now several weeks into a gruelling lockdown.

The federal government’s newly-revamped tool can lead to increased costs. Commissioner Trinidad Navarro has released a rare consumer alert regarding Medicare’s newly-updated Prescription Plan Finder. Throughout this year’s open enrollment period, which ends on December 7, the Department of Insurance has received numerous complaints about the tool and anticipated prescription drug costs. The Prescription Plan […]

The Silver Lake infusion values it at an equity value of Rs 4.90 lakh crore.

The Congress on Saturday said there was "confusion" within the central government in its fight against the novel coronavirus and wondered how would India tackle the pandemic if officials continued to speak in different voices.

This Metasploit module exploits a type confusion vulnerability found in the ActiveX component of Adobe Flash Player. This vulnerability was found exploited in the wild in November 2013. This Metasploit module has been tested successfully on IE 6 to IE 10 with Flash 11.7, 11.8 and 11.9 prior to 11.9.900.170 over Windows XP SP3 and Windows 7 SP1.

This Metasploit module exploits an issue in Google Chrome version 80.0.3987.87 (64 bit). The exploit corrupts the length of a float array (float_rel), which can then be used for out of bounds read and write on adjacent memory. The relative read and write is then used to modify a UInt64Array (uint64_aarw) which is used for read and writing from absolute memory. The exploit then uses WebAssembly in order to allocate a region of RWX memory, which is then replaced with the payload shellcode. The payload is executed within the sandboxed renderer process, so the browser must be run with the --no-sandbox option for the payload to work correctly.

PHP-Fusion CMS versions 9 through 9.03 suffer from multiple cross site scripting vulnerabilities.

PHP-Fusion version 9.03.50 suffers from an arbitrary file upload vulnerability.

PHP-Fusion version 9.03.50 suffers from a persistent cross site scripting vulnerability.

Adobe ColdFusion versions 9.0, 9.0.1, and 9.0.2 do not properly check the "rdsPasswordAllowed" field when accessing the Administrator API CFC that is used for logging in. The login function never checks if RDS is enabled when rdsPasswordAllowed="true". This means that if RDS was not configured, the RDS user does not have a password associated with their username. This means by setting rdsPasswordAllowed to "true", we can bypass the admin login to use the rdsPassword, which in most cases, is blank. These details were purchased through the Packet Storm Bug Bounty program and are being released to the community.

This Metasploit module exploits a type confusion bug in the Javascript Proxy object in WebKit. The DFG JIT does not take into account that, through the use of a Proxy, it is possible to run arbitrary JS code during the execution of a CreateThis operation. This makes it possible to change the structure of e.g. an argument without causing a bailout, leading to a type confusion (CVE-2018-4233). The type confusion leads to the ability to allocate fake Javascript objects, as well as the ability to find the address in memory of a Javascript object. This allows us to construct a fake JSCell object that can be used to read and write arbitrary memory from Javascript. The module then uses a ROP chain to write the first stage shellcode into executable memory within the Safari process and kick off its execution. The first stage maps the second stage macho (containing CVE-2017-13861) into executable memory, and jumps to its entrypoint. The CVE-2017-13861 async_wake exploit leads to a kernel task port (TFP0) that can read and write arbitrary kernel memory. The processes credential and sandbox structure in the kernel is overwritten and the meterpreter payloads code signature hash is added to the kernels trust cache, allowing Safari to load and execute the (self-signed) meterpreter payload.

Community Solar in New York has a messaging problem. It is confusing, and even some industry professionals have given up in disgust because of aggressive marketing and a lack of clarity.

EAC members have over time slowly digressed from commitments.

The language of therapy has a stranglehold on our culture. Children don’t lie anymore, they tell stories. Serial adulterers have been re-branded as sex addicts. Drunkenness is now an alcohol disorder—in fact, addiction itself is treated like a disease. Even the gross perversion of pedophilia is listed as a psychiatric disorder in the ever-expanding Diagnostic and Statistical Manual of Mental Disorders (DSM-5).

Islamabad: The cancellation of educational board exams over the coronavirus pandemic has caused confusion for candidates, Federal Minister of Education and Professional Training Shafqat Mahmood has promised to respond to questions early next week.In multiple tweets, the minister said Class 9 exam...

In a little over a year, when Americans file their 2020 tax returns, much of what is going on with coronavirus-linked stimulus payments will finally add up.

Title: Artificial Cervical Disc Surgery Cheaper Than Fusion
Category: Health News
Created: 4/29/2008 2:00:00 AM
Last Editorial Review: 4/29/2008 12:00:00 AM

Title: FDA Targets Safety Problems With Infusion Drug Pumps
Category: Health News
Created: 4/23/2010 4:10:00 PM
Last Editorial Review: 4/26/2010 12:00:00 AM

Title: Wine, Salt, and Your Heart: Confusion Abounds
Category: Health News
Created: 4/27/2011 11:00:00 AM
Last Editorial Review: 4/27/2011 12:00:00 AM

Kaposi sarcoma-associated herpesvirus (KSHV) is necessary but not sufficient for primary effusion lymphoma (PEL) development. Alterations in cellular signaling pathways are also a characteristic of PEL. Other B cell lymphomas have acquired an oncogenic mutation in the myeloid differentiation primary response 88 (MYD88) gene. The MYD88 L265P mutant results in the activation of interleukin-1 receptor associated kinase (IRAK). To probe IRAK/MYD88 signaling in PEL, we employed CRISPR/Cas9 technology to generate stable deletion clones in BCBL-1Cas9 and BC-1Cas9 cells. To look for off-target effects, we determined the complete exome of the BCBL-1Cas9 and BC-1Cas9 cells. Deletion of either MYD88, IRAK4, or IRAK1 abolished interleukin-1 beta (IL-1β) signaling; however, we were able to grow stable subclones from each population. Transcriptome sequencing (RNA-seq) analysis of IRAK4 knockout cell lines (IRAK4 KOs) showed that the IRAK pathway induced cellular signals constitutively, independent of IL-1β stimulation, which was abrogated by deletion of IRAK4. Transient complementation with IRAK1 increased NF-B activity in MYD88 KO, IRAK1 KO, and IRAK4 KO cells even in the absence of IL-1β. IL-10, a hallmark of PEL, was dependent on the IRAK pathway, as IRAK4 KOs showed reduced IL-10 levels. We surmise that, unlike B cell receptor (BCR) signaling, MYD88/IRAK signaling is constitutively active in PEL, but that under cell culture conditions, PEL rapidly became independent of this pathway.

IMPORTANCE One hundred percent of primary effusion lymphoma (PEL) cases are associated with Kaposi sarcoma-associated herpesvirus (KSHV). PEL cell lines, such as BCBL-1, are the workhorse for understanding this human oncovirus and the host pathways that KSHV dysregulates. Understanding their function is important for developing new therapies as well as identifying high-risk patient groups. The myeloid differentiation primary response 88 (MYD88)/interleukin-1 receptor associated kinase (IRAK) pathway, which has progrowth functions in other B cell lymphomas, has not been fully explored in PEL. By performing CRISPR/Cas9 knockout (KO) studies targeting the IRAK pathway in PEL, we were able to determine that established PEL cell lines can circumvent the loss of IRAK1, IRAK4, and MYD88; however, the deletion clones are deficient in interleukin-10 (IL-10) production. Since IL-10 suppresses T cell function, this suggests that the IRAK pathway may serve a function in vivo and during early-stage development of PEL.

Porcine reproductive and respiratory syndrome (PRRS) is a serious viral disease affecting the global swine industry. Its causative agent, PRRS virus (PRRSV), is an enveloped virus, and therefore membrane fusion between its envelope and host cell target membrane is critical for viral infection. Though much research has focused on PRRSV infection, the detailed mechanisms involved in its membrane fusion remain to be elucidated. In the present study, we performed confocal microscopy in combination with a constitutively active (CA) or dominant negative (DN) mutant, specific inhibitors, and small interfering RNAs (siRNAs), as well as multiple other approaches, to explore PRRSV membrane fusion. We first observed that PRRSV membrane fusion occurred in Rab11-recycling endosomes during early infection using labeled virions and subcellular markers. We further demonstrated that low pH and cathepsin E in Rab11-recycling endosomes are critical for PRRSV membrane fusion. Moreover, PRRSV glycoprotein 5 (GP5) is identified as being cleaved by cathepsin E during this process. Taken together, our findings provide in-depth information regarding PRRSV pathogenesis, which support a novel basis for the development of antiviral drugs and vaccines.

IMPORTANCE PRRS, caused by PRRSV, is an economically critical factor in pig farming worldwide. As PRRSV is a lipid membrane-wrapped virus, merging of the PRRSV envelope with the host cell membrane is indispensable for viral infection. However, there is a lack of knowledge on its membrane fusion. Here, we first explored when and where PRRSV membrane fusion occurs. Furthermore, we determined which host cell factors were involved in the process. Importantly, PRRSV GP5 is shown to be cleaved by cathepsin E during membrane fusion. Our work not only provides information on PRRSV membrane fusion for the first time but also deepens our understanding of the molecular mechanisms of PRRSV infection, which provides a foundation for future applications in the prevention and control of PRRS.

For cell entry, vaccinia virus requires fusion with the host membrane via a viral fusion complex of 11 proteins, but the mechanism remains unclear. It was shown previously that the viral proteins A56 and K2 are expressed on infected cells to prevent superinfection by extracellular vaccinia virus through binding to two components of the viral fusion complex (G9 and A16), thereby inhibiting membrane fusion. To investigate how the A56/K2 complex inhibits membrane fusion, we performed experimental evolutionary analyses by repeatedly passaging vaccinia virus in HeLa cells overexpressing the A56 and K2 proteins to isolate adaptive mutant viruses. Genome sequencing of adaptive mutants revealed that they had accumulated a unique G9R open reading frame (ORF) mutation, resulting in a single His44Tyr amino acid change. We engineered a recombinant vaccinia virus to express the G9^H44Y mutant protein, and it readily infected HeLa-A56/K2 cells. Moreover, similar to the A56 virus, the G9^H44Y mutant virus on HeLa cells had a cell fusion phenotype, indicating that G9^H44Y-mediated membrane fusion was less prone to inhibition by A56/K2. Coimmunoprecipitation experiments demonstrated that the G9^H44Y protein bound to A56/K2 at neutral pH, suggesting that the H44Y mutation did not eliminate the binding of G9 to A56/K2. Interestingly, upon acid treatment to inactivate A56/K2-mediated fusion inhibition, the G9^H44Y mutant virus induced robust cell-cell fusion at pH 6, unlike the pH 4.7 required for control and revertant vaccinia viruses. Thus, A56/K2 fusion suppression mainly targets the G9 protein. Moreover, the G9^H44Y mutant protein escapes A56/K2-mediated membrane fusion inhibition most likely because it mimics an acid-induced intermediate conformation more prone to membrane fusion.

IMPORTANCE It remains unclear how the multiprotein entry fusion complex of vaccinia virus mediates membrane fusion. Moreover, vaccinia virus contains fusion suppressor proteins to prevent the aberrant activation of this multiprotein complex. Here, we used experimental evolution to identify adaptive mutant viruses that overcome membrane fusion inhibition mediated by the A56/K2 protein complex. We show that the H44Y mutation of the G9 protein is sufficient to overcome A56/K2-mediated membrane fusion inhibition. Treatment of virus-infected cells at different pHs indicated that the H44Y mutation lowers the threshold of fusion inhibition by A56/K2. Our study provides evidence that A56/K2 inhibits the viral fusion complex via the latter’s G9 subcomponent. Although the G9^H44Y mutant protein still binds to A56/K2 at neutral pH, it is less dependent on low pH for fusion activation, implying that it may adopt a subtle conformational change that mimics a structural intermediate induced by low pH.

The entry/fusion complex (EFC) consists of 11 conserved proteins embedded in the membrane envelope of mature poxvirus particles. Poxviruses also encode proteins that localize in cell membranes and negatively regulate superinfection and syncytium formation. The vaccinia virus (VACV) A56/K2 fusion regulatory complex associates with the G9/A16 EFC subcomplex, but functional support for the importance of this interaction was lacking. Here, we describe serially passaging VACV in nonpermissive cells expressing A56/K2 as an unbiased approach to isolate and analyze escape mutants. Viruses forming large plaques in A56/K2 cells increased in successive rounds of infection, indicating the occurrence and enrichment of adaptive mutations. Sequencing of genomes of passaged and cloned viruses revealed mutations near the N terminus of the G9 open reading frame but none in A16 or other genes. The most frequent mutation was His to Tyr at amino acid 44; additional escape mutants had a His-to-Arg mutation at amino acid 44 or a duplication of amino acids 26 to 39. An adaptive Tyr-to-Cys substitution at amino acid 42 was discovered using error-prone PCR to generate additional mutations. Myristoylation of G9 was unaffected by the near-N-terminal mutations. The roles of the G9 mutations in enhancing plaque size were validated by homologous recombination. The mutants exhibited enhanced entry and spread in A56/K2 cells and induced syncytia at neutral pH in HeLa cells despite the expression of A56/K2. The data suggest that the mutations perturb the interaction of G9 with A56/K2, although some association was still detected in detergent-treated infected cell lysates.

IMPORTANCE The entry of enveloped viruses is achieved by the fusion of viral and cellular membranes, a critical step in infection that determines host range and provides targets for vaccines and therapeutics. Poxviruses encode an exceptionally large number of proteins comprising the entry/fusion complex (EFC), which enables infection of diverse cells. Vaccinia virus (VACV), the prototype member of the poxvirus family, also encodes the fusion regulatory proteins A56 and K2, which are displayed on the plasma membrane and may be beneficial by preventing reinfection and cell-cell fusion. Previous studies showed that A56/K2 interacts with the G9/A16 EFC subcomplex in detergent-treated cell extracts. Functional evidence for the importance of this interaction was obtained by serially passaging wild-type VACV in cells that are nonpermissive because of A56/K2 expression. VACV mutants with amino acid substitutions or duplications near the N terminus of G9 were enriched because of their ability to overcome the block to entry imposed by A56/K2.