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Expanded Access is a program designed to connect patients with experimental therapeutics when no other FDA-approved options are available for their given disease. In the 1980s, Dr. Thomas E Starzl, the father of modern organ transplantation, pursued the development and approval of multiple experimental immunosuppression compounds to prevent acute organ rejection in transplant patients. We will examine a historical case study of Dr. Starzl’s work with OKT-3 (muromonab) which will provide context for a discussion regarding the degree of autonomy we should grant physicians to make ethical decisions in critical Expanded Access cases, as well as the amount of oversight necessary to protect these vulnerable patients. Although this case is historical, we may gain insight into the modern problem of Expanded Access which parallels Dr. Starzl’s compassionate use requests for experimental therapies nearly three decades ago.

Expanded Access is a program designed to connect patients with experimental therapeutics when no other FDA-approved options are available for their given disease. In the 1980s, Dr. Thomas E Starzl, the father of modern organ transplantation, pursued the development and approval of multiple experimental immunosuppression compounds to prevent acute organ rejection in transplant patients. We will examine a historical case study of Dr. Starzl’s work with OKT-3 (muromonab) which will provide context for a discussion regarding the degree of autonomy we should grant physicians to make ethical decisions in critical Expanded Access cases, as well as the amount of oversight necessary to protect these vulnerable patients. Although this case is historical, we may gain insight into the modern problem of Expanded Access which parallels Dr. Starzl’s compassionate use requests for experimental therapies nearly three decades ago.

Genetically modified (GM) mice are essential tools in biomedical research. Traditional methods for generating GM mice are expensive and require specialized personnel and equipment. The use of clustered regularly interspaced short palindromic repeats (CRISPR) coupled with improved-Genome editing via Oviductal Nucleic Acids Delivery (i-GONAD) has highly increased the feasibility of producing GM mice in research laboratories. However, genetic modification in inbred mouse strains of interest such as C57BL/6 (B6) is still challenging because of their low fertility and embryo fragility. We have successfully generated multiple novel GM mouse strains in the B6 background while attempting to optimize i-GONAD. We found that i-GONAD reduced the litter size in superovulated pregnant females but did not impact pregnancy rates. Natural mating or low-hormone dose did not increase the low fertility rate observed in superovulated B6 females. However, diet enrichment had a positive effect on pregnancy success. We also optimized breeding conditions to increase the survival of small litters by co-housing i-GONAD-treated pregnant B6 females with synchronized pregnant FVB/NJ companion mothers. Thus, GM mice generation was increased by an enriched diet and shared pup rearing with highly fertile females such as FVB/NJ. In the present study, we generated 16 GM mice using a CRISPR/Cas system to target individual and multiple loci simultaneously or consecutively. We also compared homology-directed repair efficiency using different methods for LoxP insertion for conditional knockout mouse production. We found that a two-step serial LoxP insertion, in which each LoxP sequence was inserted individually in different i-GONAD procedures, was a low-risk high-efficiency method for generating floxed mice.

In this survey study of institutions across the US, marked variability in evaluation, treatment, and follow-up of adolescents 12 through 18 years of age with mRNA coronavirus disease 2019 (COVID-19) vaccine-associated myopericarditis was noted. Only one adolescent with life-threatening complications was reported, with no deaths at any of the participating institutions.

The congenital heart care community faces a myriad of public health issues that act as barriers toward optimum patient outcomes. In this article, we attempt to define advocacy and policy initiatives meant to spotlight and potentially address these challenges. Issues are organized into the following 3 key facets of our community: patient population, health care delivery, and workforce. We discuss the social determinants of health and health care disparities that affect patients in the community that require the attention of policy makers. Furthermore, we highlight the many needs of the growing adults with congenital heart disease and those with comorbidities, highlighting concerns regarding the inequities in access to cardiac care and the need for multidisciplinary care. We also recognize the problems of transparency in outcomes reporting and the promising application of telehealth. Finally, we highlight the training of providers, measures of productivity, diversity in the workforce, and the importance of patient– family centered organizations in advocating for patients. Although all of these issues remain relevant to many subspecialties in medicine, this article attempts to illustrate the unique needs of this population and highlight ways in which to work together to address important opportunities for change in the cardiac care community and beyond. This article provides a framework for policy and advocacy efforts for the next decade.

The congenital heart care community faces a myriad of public health issues that act as barriers toward optimum patient outcomes. In this article, we attempt to define advocacy and policy initiatives meant to spotlight and potentially address these challenges. Issues are organized into the following 3 key facets of our community: patient population, health care delivery, and workforce. We discuss the social determinants of health and health care disparities that affect patients in the community that require the attention of policy makers. Furthermore, we highlight the many needs of the growing adults with congenital heart disease and those with comorbidities, highlighting concerns regarding the inequities in access to cardiac care and the need for multidisciplinary care. We also recognize the problems of transparency in outcomes reporting and the promising application of telehealth. Finally, we highlight the training of providers, measures of productivity, diversity in the workforce, and the importance of patient– family centered organizations in advocating for patients. Although all of these issues remain relevant to many subspecialties in medicine, this article attempts to illustrate the unique needs of this population and highlight ways in which to work together to address important opportunities for change in the cardiac care community and beyond. This article provides a framework for policy and advocacy efforts for the next decade.

Cytomegalovirus (CMV) causes clinically important diseases in immune compromised and immune immature individuals. Based largely on work in the mouse model of murine (M)CMV, there is a consensus that myeloid cells are important for disseminating CMV from the site of infection. In theory, such dissemination should expose CMV to cell-mediated immunity and thus necessitate evasion of T cells and NK cells. However, this hypothesis remains untested. We constructed a recombinant MCMV encoding target sites for the hematopoietic specific miRNA miR-142-3p in the essential viral gene IE3. This virus disseminated poorly to the salivary gland following intranasal or footpad infections but not following intraperitoneal infection in C57BL/6 mice, demonstrating that dissemination by hematopoietic cells is essential for specific routes of infection. Remarkably, depletion of NK cells or T cells restored dissemination of this virus in C57BL/6 mice after intranasal infection, while dissemination occurred normally in BALB/c mice, which lack strong NK cell control of MCMV. These data show that cell-mediated immunity is responsible for restricting MCMV to hematopoietic cell-mediated dissemination. Infected hematopoietic cells avoided cell-mediated immunity via three immune evasion genes that modulate class I MHC and NKG2D ligands (m04, m06 and m152). MCMV lacking these 3 genes spread poorly to the salivary gland unless NK cells were depleted, but also failed to replicate persistently in either the nasal mucosa or salivary gland unless CD8+ T cells were depleted. Surprisingly, CD8+ T cells primed after intranasal infection required CD4+ T cell help to expand and become functional. Together, our data suggest that MCMV can use both hematopoietic cell-dependent and -independent means of dissemination after intranasal infection and that cell mediated immune responses restrict dissemination to infected hematopoietic cells, which are protected from NK cells during dissemination by viral immune e

We conducted a pilot study using a passive radio-wave-based home monitor in individuals with Parkinson disease (PD) with a focus on gait, home activity, and time in bed. We enrolled 7 ambulatory individuals to have the device installed in the bedroom of their homes over 8 weeks and performed standard PD assessments at baseline. We evaluated the ability of the device to objectively measure gait and time in bed and to generate novel visualizations of home activity. We captured 353 days of monitoring. Mean gait speed (0.39-0.78 m/s), time in bed per day (4.4-12.1 h), and number (1.4-5.9) and duration (15.0-49.8 min) of nightly awakenings varied substantially across and within individuals. Derived gait speed correlated well with the Movement Disorder Society-Unified Parkinson's Disease Rating Scale total (r = -0.88, p = 0.009) and motor sub-score (r = -0.95, p = 0.001). Six of the seven participants agreed that their activity was typical and indicated a willingness to continue monitoring. This technology provided promising new insights into the home activities of those with PD and may be broadly applicable to other chronic conditions., National Institute of Neurological Disorders and Stroke (Grants P20 NS092529-02, P50 NS108676-01).

The ability of deep learning (DL) to improve the practice of medicine and its clinical outcomes faces a looming obstacle: model interpretation. Without description of how outputs are generated, a collaborating physician can neither resolve when the model's conclusions are in conflict with his or her own, nor learn to anticipate model behavior. Current research aims to interpret networks that diagnose ECG recordings, which has great potential impact as recordings become more personalized and widely deployed. A generalizable impact beyond ECGs lies in the ability to provide a rich test-bed for the development of interpretive techniques in medicine. Interpretive techniques for Deep Neural Networks (DNNs), however, tend to be heuristic and observational in nature, lacking the mathematical rigor one might expect in the analysis of math equations. The motivation of this paper is to offer a third option, a scientific approach. We treat the model output itself as a phenomenon to be explained through component parts and equations governing their behavior. We argue that these component parts should also be "black boxes" --additional targets to interpret heuristically with clear functional connection to the original. We show how to rigorously factor a DNN into a hierarchical equation consisting of black box variables. This is not a subdivision into physical parts, like an organism into its cells; it is but one choice of an equation into a collection of abstract functions. Yet, for DNNs trained to identify normal ECG waveforms on PhysioNet 2017 Challenge data, we demonstrate this choice yields interpretable component models identified with visual composite sketches of ECG samples in corresponding input regions. Moreover, the recursion distills this interpretation: additional factorization of component black boxes corresponds to ECG partitions that are more morphologically pure.

Not only are Deep Neural Networks (DNNs) black box models, but also we frequently conceptualize them as such. We lack good interpretations of the mechanisms linking inputs to outputs. Therefore, we find it difficult to analyze in human-meaningful terms (1) what the network learned and (2) whether the network learned. We present a hierarchical decomposition of the DNN discrete classification map into logical (AND/OR) combinations of intermediate (True/False) classifiers of the input. Those classifiers that can not be further decomposed, called atoms, are (interpretable) linear classifiers. Taken together, we obtain a logical circuit with linear classifier inputs that computes the same label as the DNN. This circuit does not structurally resemble the network architecture, and it may require many fewer parameters, depending on the configuration of weights. In these cases, we obtain simultaneously an interpretation and generalization bound (for the original DNN), connecting two fronts which have historically been investigated separately. Unlike compression techniques, our representation is. We motivate the utility of this perspective by studying DNNs in simple, controlled settings, where we obtain superior generalization bounds despite using only combinatorial information (e.g. no margin information). We demonstrate how to "open the black box" on the MNIST dataset. We show that the learned, internal, logical computations correspond to semantically meaningful (unlabeled) categories that allow DNN descriptions in plain English. We improve the generalization of an already trained network by interpreting, diagnosing, and replacing components the logical circuit that is the DNN.

Lester O Hosten,1 Christopher Snyder2 1Clinical and Medical Affairs, Bausch + Lomb, Bridgewater, NJ, USA; 2Regional Director Medical Science, Bausch + Lomb, Birmingham, AL, USACorrespondence: Lester O HostenClinical and Medical Affairs, Bausch + Lomb, 400 Somerset Corporate Boulevard, Bridgewater, NJ 08807, USATel +1 (908) 541-5449Email lester.hosten@bausch.comAbstract: To manage ocular redness effectively, health-care practitioners require an understanding of the pathophysiology, clinical features and differential diagnosis of ocular redness, as well as comprehensive knowledge of medical therapies available and their pharmacologic properties. This review aims to provide a clinically relevant summary of the current literature on the mechanism of action, efficacy, and safety of current over-the-counter (OTC) decongestants available for reduction of ocular redness due to minor irritations. Currently marketed OTC products indicated for such use in the United States include topical solutions of tetrahydrozoline 0.05%, naphazoline 0.012% to 0.03%, and brimonidine 0.025%. All 3 agents are adrenergic receptor agonists but vary in their receptor-binding profiles: tetrahydrozoline is a selective α 1 receptor agonist; naphazoline is a mixed α 1/α 2 receptor agonist; and brimonidine is a selective α 2 receptor agonist. These OTC decongestants produce vasoconstriction of conjunctival blood vessels, which results in a rapid reduction in ocular redness. In general, ocular adverse events reported in published studies of these OTC decongestants were minimal, mild, and transient, with no significant adverse systemic effects. However, ocular decongestants with α 1-adrenergic receptor agonist activity can be associated with loss of effectiveness with continued use (ie, tachyphylaxis) and rebound redness upon treatment discontinuation. In clinical trials of the selective α 2-adrenergic receptor agonist brimonidine 0.025%, tach

Cytomegalovirus (CMV) is a herpesvirus that establishes a persistent, but generally asymptomatic, infection in most people in the world. However, CMV drives and sustains extremely large numbers of antigen-specific T cells and is, therefore, emerging as an exciting platform for vaccines against infectious diseases and cancer. Indeed, pre-clinical data strongly suggest that CMV-based vaccines can sustain protective CD8+ T cell and antibody responses. In the context of vaccines for infectious diseases, substantial pre-clinical studies have elucidated the effcacy and protective mechanisms of CMV-based vaccines, including in non-human primate models of various infections. In the context of cancer vaccines, however, much less is known and only very early studies in mice have been conducted. To develop CMV-based cancer vaccines further, it will be critical to better understand the complex interaction of CMV and cancer. An array of evidence suggests that naturally-acquired human (H)CMV can be detected in cancers, and it has been proposed that HCMV may promote tumor growth. This would obviously be a concern for any therapeutic cancer vaccines. In experimental models, CMV has been shown to play both positive and negative roles in tumor progression, depending on the model studied. However, the mechanisms are still largely unknown. Thus, more studies assessing the interaction of CMV with the tumor microenvironment are needed. This review will summarize the existing literature and major open questions about CMV-based vaccines for cancer, and discuss our hypothesis that the balance between pro-tumor and anti-tumor effects driven by CMV depends on the location and the activity of the virus in the lesion.

The springtime sublimation process of Mars' southern seasonal polar CO$_2$ ice cap features dark fan-shaped deposits appearing on the top of the thawing ice sheet. The fan material likely originates from the surface below the ice sheet, brought up via CO$_2$ jets breaking through the seasonal ice cap. Once the dust and dirt is released into the atmosphere, the material may be blown by the surface winds into the dark streaks visible from orbit. The location, size and direction of these fans record a number of parameters important to quantifying seasonal winds and sublimation activity, the most important agent of geological change extant on Mars. We present results of a systematic mapping of these south polar seasonal fans with the Planet Four online citizen science project. Planet Four enlists the general public to map the shapes, directions, and sizes of the seasonal fans visible in orbital images. Over 80,000 volunteers have contributed to the Planet Four project, reviewing 221 images, from Mars Reconnaissance Orbiter's HiRISE (High Resolution Imaging Science Experiment) camera, taken in southern spring during Mars Years 29 and 30. We provide an overview of Planet Four and detail the processes of combining multiple volunteer assessments together to generate a high fidelity catalog of $\sim$ 400,000 south polar seasonal fans. We present the results from analyzing the wind directions at several locations monitored by HiRISE over two Mars years, providing new insights into polar surface winds., Comment: 75 pages, 46 figures, minor revisions discovered in proof, accepted and in press at Icarus

The springtime sublimation process of Mars' southern seasonal polar CO$_2$ ice cap features dark fan-shaped deposits appearing on the top of the thawing ice sheet. The fan material likely originates from the surface below the ice sheet, brought up via CO$_2$ jets breaking through the seasonal ice cap. Once the dust and dirt is released into the atmosphere, the material may be blown by the surface winds into the dark streaks visible from orbit. The location, size and direction of these fans record a number of parameters important to quantifying seasonal winds and sublimation activity, the most important agent of geological change extant on Mars. We present results of a systematic mapping of these south polar seasonal fans with the Planet Four online citizen science project. Planet Four enlists the general public to map the shapes, directions, and sizes of the seasonal fans visible in orbital images. Over 80,000 volunteers have contributed to the Planet Four project, reviewing 221 images, from Mars Reconnaissance Orbiter's HiRISE (High Resolution Imaging Science Experiment) camera, taken in southern spring during Mars Years 29 and 30. We provide an overview of Planet Four and detail the processes of combining multiple volunteer assessments together to generate a high fidelity catalog of $\sim$ 400,000 south polar seasonal fans. We present the results from analyzing the wind directions at several locations monitored by HiRISE over two Mars years, providing new insights into polar surface winds., Comment: 75 pages, 46 figures, minor revisions discovered in proof, accepted and in press at Icarus

In 2013, Harford County adopted a Bicycle and Pedestrian Master Plan that establishes goals and priorities for improving the County’s bicycle and pedestrian networks and connecting to key destinations throughout the County. In 2018, Harford County engaged with the Bachelor of Landscape Architecture Capstone Studio at the University of Maryland to develop a more comprehensive plan that would include an overall strategy and suite of best practices to guide the development, implementation and expansion of the County’s bicycle network. During the studio’s first phase, students reviewed the existing plan, supporting materials, precedents of bicycle trails and Complete Streets policies, and met with planning officials to evaluate information gaps and missing links and to provide recommendations for developing links. Working in design teams, students addressed the priority connections between the County and the Town of Bel Air. The teams also identified potential connections to or expansions of the green infrastructure network and opportunities for recreational tourism. During the studio’s second phase, students undertook a more detailed analysis of the Town of Bel Air’s 2013 Bicycle and Pedestrian Plan to advance the proposed segments that connect to the County’s Plan. County-identified priorities include Macphail Road, MD Route 24, and the connection between downtown Bel Air and Harford Community College. The teams field verified the concepts for proposed routes. Standards for trail designs and markers were developed and marker locations were identified.

Even though Deep Neural Networks (DNNs) are widely celebrated for their practical performance, they possess many intriguing properties related to depth that are difficult to explain both theoretically and intuitively. Understanding how weights in deep networks coordinate together across layers to form useful learners has proven challenging, in part because the repeated composition of nonlinearities has proved intractable. This paper presents a reparameterization of DNNs as a linear function of a feature map that is locally independent of the weights. This feature map transforms depth-dependencies into simple tensor products and maps each input to a discrete subset of the feature space. Then, using a max-margin assumption, the paper develops a sample compression representation of the neural network in terms of the discrete activation state of neurons induced by s ``support vectors". The paper shows that the number of support vectors s relates with learning guarantees for neural networks through sample compression bounds, yielding a sample complexity of O(ns/epsilon) for networks with n neurons. Finally, the number of support vectors s is found to monotonically increase with width and label noise but decrease with depth., Comment: 15 pages, 10 figures

The sepsis-induced cytokine storm leads to severe lymphopenia and reduced effector capacity of remaining/surviving cells. This results in a prolonged state of immunoparalysis, that contributes to enhanced morbidity/mortality of sepsis survivors upon secondary infection. The impact of sepsis on several lymphoid subsets has been characterized, yet its impact on NK-cells remains underappreciated-despite their critical role in controlling infection(s). Here, we observed numerical loss of NK-cells in multiple tissues after cecal-ligation-and-puncture (CLP)-induced sepsis. To elucidate the sepsis-induced lesions in surviving NK-cells, transcriptional profiles were evaluated and indicated changes consistent with impaired effector functionality. A corresponding deficit in NK-cell capacity to produce effector molecules following secondary infection and/or cytokine stimulation (IL-12,IL-18) further suggested a sepsis-induced NK-cell intrinsic impairment. To specifically probe NK-cell receptor-mediated function, the activating Ly49H receptor, that recognizes the murine cytomegalovirus (MCMV) m157 protein, served as a model receptor. Although relative expression of Ly49H receptor did not change, the number of Ly49H+ NK-cells in CLP hosts was reduced leading to impaired in vivo cytotoxicity and the capacity of NK-cells (on per-cell basis) to perform Ly49H-mediated degranulation, killing, and effector molecule production in vitro was also severely reduced. Mechanistically, Ly49H adaptor protein (DAP12) activation and clustering, assessed by TIRF microscopy, was compromised. This was further associated with diminished AKT phosphorylation and capacity to flux calcium following receptor stimulation. Importantly, DAP12 overexpression in NK-cells restored Ly49H/D receptors-mediated effector functions in CLP hosts. Finally, as a consequence of sepsis-dependent numerical and functional lesions in Ly49H+ NK-cells, host capacity to control MCMV infection was significantly impaired. Import

Head and neck squamous cell carcinoma (HNSCC) is the 6th most common human cancer and affects approximately 50,000 new patients every year in the US. The major risk factors for HNSCC are tobacco and alcohol consumption as well as oncogenic HPV infections. Despite advances in therapy, the overall survival rate for all-comers is only 50%. Understanding the biology of HNSCC is crucial to identifying new biomarkers, implementing early diagnostic approaches and developing novel therapies. As in several other cancers, HNSCC expresses elevated levels of MCT4, a member of the SLC16 family of monocarboxylate transporters. MCT4 is a H+-linked lactate transporter which functions to facilitate lactate efflux from highly glycolytic cells. High MCT4 levels in HNSCC have been associated with poor prognosis, but the role of MCT4 in the development and progression of this cancer is still poorly understood. In this study, we used 4-nitroquinoline-1-oxide (4NQO) to induce oral cancer in MCT4-/- and wild type littermates, recapitulating the disease progression in humans. Histological analysis of mouse tongues after 23 weeks of 4NQO treatment showed that MCT4-/- mice developed significantly fewer and less extended invasive lesions than wild type. In mice, as in human samples, MCT4 was not expressed in normal oral mucosa but was detected in the transformed epithelium. In the 4NQO treated mice we detected MCT4 in foci of the basal layer undergoing transformation, and progressively in areas of carcinoma in situ and invasive carcinomas. Moreover, we found MCT4 positive macrophages within the tumor and in the stroma surrounding the lesions in both human samples of HNSCC and in the 4NQO treated animals. The results of our studies showed that MCT4 could be used as an early diagnostic biomarker of HNSCC. Our finding with the MCT4-/- mice suggest MCT4 is a driver of progression to oral squamous cell cancer and MCT4 inhibitors could have clinical benefits for preventing invasive HNSCC. © 2018 Bis

We propose an adversarial training procedure for learning a causal implicit generative model for a given causal graph. We show that adversarial training can be used to learn a generative model with true observational and interventional distributions if the generator architecture is consistent with the given causal graph. We consider the application of generating faces based on given binary labels where the dependency structure between the labels is preserved with a causal graph. This problem can be seen as learning a causal implicit generative model for the image and labels. We devise a two-stage procedure for this problem. First we train a causal implicit generative model over binary labels using a neural network consistent with a causal graph as the generator. We empirically show that WassersteinGAN can be used to output discrete labels. Later, we propose two new conditional GAN architectures, which we call CausalGAN and CausalBEGAN. We show that the optimal generator of the CausalGAN, given the labels, samples from the image distributions conditioned on these labels. The conditional GAN combined with a trained causal implicit generative model for the labels is then a causal implicit generative model over the labels and the generated image. We show that the proposed architectures can be used to sample from observational and interventional image distributions, even for interventions which do not naturally occur in the dataset.

Human cytomegalovirus (HCMV) is a ubiquitous virus that causes chronic infection and, thus, is one of the most common infectious complications of immune suppression. Adoptive transfer of HCMV-specific T cells has emerged as an effective method to reduce the risk for HCMV infection and/or reactivation by restoring immunity in transplant recipients. However, the CMV-specific CD8(+) T cell response is comprised of a heterogenous mixture of subsets with distinct functions and localization, and it is not clear if current adoptive immunotherapy protocols can reconstitute the full spectrum of CD8(+) T cell immunity. The aim of this review is to briefly summarize the role of these T cell subsets in CMV immunity and to describe how current adoptive immunotherapy practices might affect their reconstitution in patients. The bulk of the CMV-specific CD8(+) T cell population is made up of terminally differentiated effector T cells with immediate effector function and a short life span. Self-renewing memory T cells within the CMV-specific population retain the capacity to expand and differentiate upon challenge and are important for the long-term persistence of the CD8(+) T cell response. Finally, mucosal organs, which are frequent sites of CMV reactivation, are primarily inhabited by tissue-resident memory T cells, which do not recirculate. Future work on adoptive transfer strategies may need to focus on striking a balance between the formation of these subsets to ensure the development of long lasting and protective immune responses that can access the organs affected by CMV disease.

OBJECTIVES Calculation of accurate T1 relaxivity (r1) values for gadolinium-based magnetic resonance contrast agents (GBCAs) is a complex process. As such, often referenced r1 values for the GBCAs at 1.5 T, 3 T, and 7 T are based on measurements obtained in media that are not clinically relevant, derived from only a small number of concentrations, or available for only a limited number of GBCAs. This study derives the r1 values of the 8 commercially available GBCAs in human whole blood at 1.5 T, 3 T, and 7 T. MATERIALS AND METHODS Eight GBCAs were serially diluted in human whole blood, at 7 concentrations from 0.0625 to 4 mM. A custom-built phantom held the dilutions in air-tight cylindrical tubes maintained at 37 ± 0.5°C by a heat-circulating system. Images were acquired using inversion recovery sequences with inversion times from 30 milliseconds to 10 seconds at 1.5 T and 3 T as well as 60 milliseconds to 5 seconds at 7 T. A custom MATLAB program was used to automate signal intensity measurements from the images acquired of the phantom. SigmaPlot was used to calculate T1 relaxation times and, finally, r1. RESULTS Measured r1 values in units of s[BULLET OPERATOR]mM at 1.5 T (3 T/7 T) were 3.9 ± 0.2 (3.4 ± 0.4/2.8 ± 0.4) for Gd-DOTA, 4.6 ± 0.2 (4.5 ± 0.3/4.2 ± 0.3) for Gd-DO3A-butrol, 4.3 ± 0.4 (3.8 ± 0.2/3.1 ± 0.4) for Gd-DTPA, 6.2 ± 0.5 (5.4 ± 0.3/4.7 ± 0.1) for Gd-BOPTA, 4.5 ± 0.1 (3.9 ± 0.2/3.7 ± 0.2) for Gd-DTPA-BMA, 4.4 ± 0.2 (4.2 ± 0.2/4.3 ± 0.2) for Gd-DTPA-BMEA, 7.2 ± 0.2 (5.5 ± 0.3/4.9 ± 0.1) for Gd-EOB-DTPA, and 4.4 ± 0.6 (3.5 ± 0.6/3.4 ± 0.1) for Gd-HP-DO3A. The agents can be stratified by relaxivity, with a significant additional dependency on field strength. CONCLUSIONS This report quantifies, for the first time, T1 relaxivity for all 8 gadolinium chelates in common clinical use worldwide, at current relevant field strengths, in human whole blood at physiological temperature (37°C). The measured r1 values differ to a small degree from previously publis

Cytomegalovirus (CMV) is a herpesvirus that persists for life and maintains extremely large numbers of T cells with select specificities in circulation. However, it is unknown how viral persistence impacts T cell populations in mucosal sites. We found that many murine (M)CMV-specific CD8s in mucosal tissues became resident memory T cells (TRM). These cells adopted an intraepithelial localization in the salivary gland that correlated with, but did not depend on, expression of the integrin CD103. MCMV-specific TRM cells formed early after infection, and spleen-localized cells had reduced capacities to become TRM at late times. Surprisingly, however, small numbers of new TRM cells were formed from the circulating pool throughout infection, favoring populations maintained at high levels in the blood and shifting the immunodominance within the TRM populations over time. These data show that mucosal TRM populations can be dynamically maintained by a persistent infection.

Reconstitution of CMV-specific immunity after transplant remains a primary clinical objective to prevent CMV disease, and adoptive immunotherapy of CMV-specific T cells can be an effective therapeutic approach. Because of viral persistence, most CMV-specific CD8(+) T cells become terminally differentiated effector phenotype CD8(+) T cells (TEFF). A minor subset retains a memory-like phenotype (memory phenotype CD8(+) T cells [TM]), but it is unknown whether these cells retain memory function or persist over time. Interestingly, recent studies suggest that CMV-specific CD8(+) T cells with different phenotypes have different abilities to reconstitute sustained immunity after transfer. The immunology of human CMV infections is reflected in the murine CMV (MCMV) model. We found that human CMV- and MCMV-specific T cells displayed shared genetic programs, validating the MCMV model for studies of CMV-specific T cells in vivo. The MCMV-specific TM population was stable over time and retained a proliferative capacity that was vastly superior to TEFF. Strikingly, after transfer, TM established sustained and diverse T cell populations even after multiple challenges. Although both TEFF and TM could protect Rag(-/-) mice, only TM persisted after transfer into immune replete, latently infected recipients and responded if recipient immunity was lost. Interestingly, transferred TM did not expand until recipient immunity was lost, supporting that competition limits the Ag stimulation of TM. Ultimately, these data show that CMV-specific TM retain memory function during MCMV infection and can re-establish CMV immunity when necessary. Thus, TM may be a critical component for consistent, long-term adoptive immunotherapy success.

Several low-grade persistent viral infections induce and sustain very large numbers of virus-specific effector T cells. This was first described as a response to cytomegalovirus (CMV), a herpesvirus that establishes a life-long persistent/latent infection, and sustains the largest known effector T cell populations in healthy people. These T cells remain functional and traffic systemically, which has led to the recent exploration of CMV as a persistent vaccine vector. However, the maintenance of this remarkable response is not understood. Current models propose that reservoirs of viral antigen and/or latently infected cells in lymph nodes stimulate T cell proliferation and effector differentiation, followed by migration of progeny to non-lymphoid tissues where they control CMV reactivation. We tested this model using murine CMV (MCMV), a natural mouse pathogen and homologue of human CMV (HCMV). While T cells within draining lymph nodes divided at a higher rate than cells elsewhere, antigen-dependent proliferation of MCMV-specific effector T cells was observed systemically. Strikingly, inhibition of T cell egress from lymph nodes failed to eliminate systemic T cell division, and did not prevent the maintenance of the inflationary populations. In fact, we found that the vast majority of inflationary cells, including most cells undergoing antigen-driven division, had not migrated into the parenchyma of non-lymphoid tissues but were instead exposed to the blood supply. Indeed, the immunodominance and effector phenotype of inflationary cells, both of which are primary hallmarks of memory inflation, were largely confined to blood-localized T cells. Together these results support a new model of MCMV-driven memory inflation in which most immune surveillance occurs in circulation, and in which most inflationary effector T cells are produced in response to viral antigen presented by cells that are accessible to the blood supply.

Cytomegalovirus (CMV) is a herpesvirus that establishes life-long latency in 60-80% of Americans. Constant immune surveillance is necessary to prevent viral reactivation from latency and results in the accumulation of functional CMV-specific CD8+ T cells (CD8s) over time, a process termed memory inflation. As such, CMV reactivations remain a clinical concern for immunosuppressed patients and reconstituting CMV immunity is critical for the long-term prevention of CMV disease. Understanding the maintenance of memory inflation may reveal novel approaches to restore CMV immunity. Previous work has shown that the majority of inflationary CD8s express a terminally-differentiated “effector” (TEFF) phenotype, have a short half-life and appear unable to sustain long-term CMV immunity. Interestingly, inflationary populations also include a minor subset of CD8s that express a “memory” phenotype (TM).

BACKGROUND: Murine cytomegalovirus (MCMV) is increasingly used as an infectious model to investigate host-pathogen interactions in mice. Detailed methods have been published for using primary murine embryonic fibroblasts (MEFs) for preparing stocks and determining viral titers of MCMV. For determining the titer of MCMV by plaque assay, these methods rely on a high viscosity media that restricts viral spreading through the supernatant of the culture, but is also usually too viscous to pipet. Moreover, MEFs must be repeatedly generated and can vary widely from batch-to-batch in purity, proliferation rates, and the development of senescence. In contrast, the M2-10B4 bone marrow stromal cell line (ATCC # CRL-1972), which is also permissive for MCMV, has been reported to produce high-titer stocks of MCMV and has the considerable advantages of growing rapidly and consistently. However, detailed methods using these cells have not been published. METHODS: We modified existing protocols to use M2-10B4 cells for measuring MCMV titers by plaque assay. RESULTS: We found that MCMV plaques could be easily resolved on monolayers of M2-10B4 cells. Moreover, plaques formed normally even when cultures of M2-10B4 cells were less than 50% confluent on the day of infection, as long as we also used a reduced viscosity overlay. CONCLUSIONS: Overall, our protocol enabled us to use a consistent cell line to assess viral titers, rather than repeatedly producing primary MEFs. It also allowed us to start the assay with 4-fold fewer cells than would be required to generate a confluent monolayer, reducing the lead-time prior to the start of the assay. Finally, the reduced viscosity CMC could be handled by pipet and did not need to be pre-mixed with media, thus increasing its shelf-life and ease-of-use. We describe our results here, along with detailed protocols for the use of the M2-10B4 cell lines to determine the titer and grow stocks of MCMV.

The unique ability of CMV to drive the expansion of virus-specific T cell populations during the course of a lifelong, persistent infection has generated interest in the virus as a potential vaccine strategy. When designing CMV-based vaccine vectors to direct immune responses against HIV or tumor Ags, it becomes important to understand how and why certain CMV-specific populations are chosen to inflate over time. To investigate this, we designed recombinant murine CMVs (MCMVs) encoding a SIINFEKL-enhanced GFP fusion protein under the control of endogenous immediate early promoters. When mice were infected with these viruses, T cells specific for the SIINFEKL epitope inflated and profoundly dominated T cells specific for nonrecombinant (i.e., MCMV-derived) Ags. Moreover, when the virus encoded SIINFEKL, T cells specific for nonrecombinant Ags displayed a phenotype indicative of less frequent exposure to Ag. The immunodominance of SIINFEKL-specific T cells could not be altered by decreasing the number of SIINFEKL-specific cells available to respond, or by increasing the number of cells specific for endogenous MCMV Ags. In contrast, coinfection with viruses expressing and lacking SIINFEKL enabled coinflation of T cells specific for both SIINFEKL and nonrecombinant Ags. Because coinfection allows presentation of SIINFEKL and MCMV-derived Ags by different cells within the same animal, these data reveal that competition for, or availability of, Ag at the level of the APC determines the composition of the inflationary response to MCMV. SIINFEKL's strong affinity for H-2K(b), as well as its early and abundant expression, may provide this epitope's competitive advantage.

Cytomegalovirus (CMV) establishes a life-long, persistent/latent infection. Continuous immune surveillance by CMV-specific CD8+ T cells results in their accumulation over time, a process called “memory inflation”. These “inflationary” T cells migrate systemically and comprise the largest T cell populations in humans, making CMV-based vaccines attractive. Why T cells specific for only some CMV-derived epitopes undergo inflation is unclear. We found previously that recombinant murine (M)CMV expressing the SIINFEKL peptide only induced inflation of SIINFEKL-specific T cells, in contrast to wild-type MCMV infection. We show here that: Co-infecting mice with viruses expressing and lacking SIINFEKL enabled T cells with multiple specificities to inflate. Adoptively transferred OT-I T cells, which have a high affinity for the SIINFEKL peptide, were preferentially enriched shortly after infection. Adoptively transferred OT-I T cells inhibited the inflation of host-derived T cells specific for SIINFEKL. We observed sporadic, late rejection of OT-Is as a result of minor histocompatibility differences between donor and recipients. Only such rejection enabled host-derived T cells to inflate robustly.

The Large Civil Tiltrotor (LCTR) was developed as part of the NASA Heavy Lift Rotorcraft Systems Investigation in order to establish a consistent basis for evaluating the benefits of advanced technology for large tiltrotors. The concept has since evolved into the second-generation LCTR2, designed to carry 90 passengers for 1,000 nm at 300 knots, with vertical takeoff and landing. This paper examines the impact of advanced propulsion system concepts on LCTR2 sizing. Two concepts were studied: an advanced, single-speed engine with a conventional power turbine layout (Advanced Conventional Engine, or ACE), and a variable-speed power turbine engine (VSPT). The ACE is the lighter engine, but requires a multi-speed (shifting) gearbox, whereas the VSPT uses a lighter, fixed-ratio gearbox. The NASA Design and Analysis of Rotorcraft (NDARC) design code was used to study the trades between rotor and engine efficiency and weight. Rotor performance was determined by Comprehensive Analytical Model of Rotorcraft Aerodynamics and Dynamics (CAMRAD II), and engine performance was estimated with the Numerical Propulsion System Simulation (NPSS). Design trades for the ACE vs. VSPT are presented in terms of vehicle weight empty for variations in mission altitude and range; the effect of different One Engine Inoperative (OEI) criteria are also examined. Because of its strong effect on gearbox weight and on both rotor and engine efficiency, rotor speed was chosen as the reference design variable for comparing design trades. The two propulsion concepts had nearly identical vehicle weights and mission fuel consumption, and their relative advantages varied little with cruise altitude, mission range, or OEI criteria; high cruise altitude and low cruise tip speed were beneficial for both concepts., Presented at the American Helicopter Society Future Vertical Lift Aircraft Design Conference, San Francisco, CA, January 18-20, 2012.

Honors programs thrive in an environment of pedagogic freedom. This freedom extends to our honors students as they explore topics for projects and theses and engage in much more independent research than the average undergraduate. Honors programs should also be havens for faculty to experiment with new ideas for courses and co-curricular activities. Freed from large lecture halls and department politics, faculty who teach in an honors program often find themselves wandering over to the honors facilities to hang out with students or going off on honors-sponsored adventures. Thus academic freedom also often leads to a stronger sense of community. However, as the corporate, managerial model encroaches on the modern university, both academic freedom and the community of scholars are under threat, and honors administrators must find a way to preserve what makes their programs unique. Universities used to generate new ideas and create models that were adopted by those outside the ivory tower, from art and entertainment to industry and politics. However, the modern university, perhaps lacking its old confidence, turns again and again to the corporate world for many of its practices, including so-called accountability. Politicians, claiming to speak for the “consumers” of higher education who spend ever-increasing sums for college tuition, have in many cases required colleges and universities that receive state and federal funding, which means just about every institution of higher learning, to show “transparency and accountability,” and the schools, urged by accreditation agencies, have decided that “assessment of student learning” is the best response to critics and consumers alike. Through reaccreditation, budgeting decisions, curriculum approval and other means, university administrators have exerted pressure upon deans, department chairs, and individual faculty members to “embrace the culture of assessment.” In our previous article for JNCHC, we questioned the validity

Cytomegalovirus (CMV) is a β-herpesvirus that establishes a lifelong latent or persistent infection. A hallmark of chronic CMV infection is the lifelong persistence of large numbers of virus-specific CD8+ effector/effector memory T cells, a phenomenon called "memory inflation". How the virus continuously stimulates these T cells without being eradicated remains an enigma. The prevailing view is that CMV establishes a low grade "smoldering" infection characterized by tiny bursts of productive infection which are rapidly extinguished, leaving no detectable virus but replenishing the latent pool and leaving the immune system in a highly charged state. However, since abortive reactivation with limited viral gene expression is known to occur commonly, we investigated the necessity for virus reproduction in maintaining the inflationary T cell pool. We inhibited viral replication or spread in vivo using two different mutants of murine CMV (MCMV). First, famcyclovir blocked the replication of MCMV encoding the HSV Thymidine Kinase gene, but had no impact on the CD8+ T cell memory inflation once the infection was established. Second, MCMV that lacks the essential glycoprotein L, and thus is completely unable to spread from cell to cell, also drove memory inflation if the virus was administered systemically. Our data suggest that CMV which cannot spread from the cells it initially infects can repeatedly generate viral antigens to drive memory inflation without suffering eradication of the latent genome pool.

Chronic infections have been a major topic of investigation in recent years, but the mechanisms that dictate whether or not a pathogen is successfully controlled are incompletely understood. Cytomegalovirus (CMV) is a herpesvirus that establishes a persistent infection in the majority of people in the world. Like other herpesviruses, CMV is well controlled by an effective immune response and induces little, if any, pathology in healthy individuals. However, controlling CMV requires continuous immune surveillance, and thus, CMV is a significant cause of morbidity and death in immune-compromised individuals. T cells in particular play an important role in controlling CMV and both CD4(+) and CD8(+) CMV-specific T cells are essential. These virus-specific T cells persist in exceptionally large numbers during the infection, traffic into peripheral tissues and remain functional, making CMV an attractive vaccine vector for driving "CMV-like" T cell responses against recombinant antigens of choice. However, the mechanisms by which these T cells persist and differentiate while remaining functional are still poorly understood, and we have no means to promote their development in immune-compromised patients at risk for CMV disease. In this review, I will briefly summarize our current knowledge of CMV-specific CD8(+) T cells and propose a mechanism that may explain their maintenance and preservation of function during chronic infection.

In its ideal form, systematic assessment is a legitimate way for honors programs and colleges to gauge strengths and weaknesses, measure the effect of various learning environments, and evoke positive institutional change based on objective, empirical data. Such assessment can take two main forms. Programmatic assessment (also known as program evaluation) is an extremely useful tool for gathering evidence and evaluating whether an honors program embodies the NCHC’s basic characteristics (Sederberg 159) and/or meets its own institutional goals, e.g., higher rates of retention, graduation, graduate/professional school acceptance, and successful competition for national fellowships. Furthermore, as Otero and Spurrier argue (5), this oftenrequired process can offer honors programs a way to improve, tailor the assessment mechanism, demonstrate program strengths, and garner financial support. Like it or not, in these competitive times programmatic assessment has become a part of American higher education, and honors programs or colleges that do not engage in it, or at least shape it to their own purpose and design, risk alienating accountability-driven entities on and off campus. While honors programs are certainly not immune to such self-interested concerns, our true bottom line is providing students with an enriched education that cultivates learning at the very highest scholarly levels. To this end, the second main type of assessment, learning outcomes assessment, attempts to measure what college students learn as a result of participation in honors and also to distinguish the unique characteristics of an honors education. This essay will focus on the second type, highlighting some limitations to the assessment of learning in honors. First, we will examine limitations in the methodology and logic of learning assessment from a behavioral science perspective, raising concerns about what we are truly measuring and how we are evaluating, interpreting, and applying this inf

The term fog of war is often associated with the commander s lack of clear information on the battlefield. War is inherently volatile, uncertain, complex, and ambiguous. For this condition, contemporary U.S. military usage offers the acronym VUCA. 1 Compounding the fog of war on the modern battlefield is the high tempo of operations or speed sought by commanders to overwhelm and defeat the enemy. This case study proposes that the use of sense and respond (S&R) logistics during Operation Iraqi Freedom (OIF-I) would have provided logisticians critical decisionmaking information (situational awareness) thereby reducing the fog of war and facilitating more efficient and responsive support to the warfighter. In drawing this conclusion, the following study analyzes the events of OIF-I, citing logistical lessons learned and difficulties experienced, and offers suggestions to reduce those challenges. The implementation of S&R logistics will shape future joint logistics requirements while driving changes in joint doctrine and how we support the operational environment. Current efforts under the Forces Transformation and Resources Office (formerly the Office of Force Transformation) and the Program Manager, Light Armored Vehicle (PMLAV) pave a path for S & R logistics implementation within the military. Lastly, the study highlights the current Marine Corps logistics operations in Iraq and offers some insight into the future. An initial overview examines the events that led to the overthrow of Saddam Hussein.

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Economics, 1994., Includes bibliographical references., by Christopher M. Snyder., Ph.D.

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Economics, 1994., Includes bibliographical references., by Christopher M. Snyder., Ph.D.

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Economics, 1994., Includes bibliographical references., by Christopher M. Snyder., Ph.D.

We consider a setting in which the buyer's ability to hold up a seller's investment is so severe that there is no investment in equilibrium of the static game typically analyzed. We show that there exists an equilibrium of a related dynamic game generating positive investment. The seller makes a sequence of gradually smaller investments, each repaid by the buyer under the threat of losing further seller investment. As modeled frictions converge to zero, the equilibrium outcome converges to the first best. We draw connections between our work and the growing literature on gradualism in public good contribution games and bargaining games.

The potential of monoclonal antibodies, (mAbs), for use in therapeutic and diagnostic applications has not been fully realized in part due to counter-immune responses that often arise in patient recipients of mAb. A growing research effort to "humanize" mAb has focused primarily on the structure or sequence of the antibody variable (V) region domains. However, these approaches may ultimately suffer, as they overlook the requirement of T cell help for the immune counter-reaction and the potential of somatic hypermutation and V-D-J recombination to generate target T cell epitopes within mAb V regions. My approach focuses on this issue. In order to understand seine basic principals concerning anti-immunoglobulin immune responses, I have developed a panal of T cell hybridomas, new transgenic mice and a tetrameric staining reagent. Studies with these tools strongly support our basic hypothesis that T cells are tolerant of endogenous immunoglobulin-derived diversity. I have also obtained a panal of T cell hybridomas that are specific for the CDR3 region of a monoclonal antibody supporting our hypothesis that junctional diversity may provide a source of T cell epitopes within a monoclonal antibody. Finally, I have addressed the global nature of T cell responses to junctional diversity with an adoptove transfer system., The original document contains color images. All DTIC reproductions will be in black and white.

The potential of monoclonal antibodies, (mAbs), for use in therapeutic and diagnostic applications has not been fully realized in part due to counter-immune responses that often arise in patient recipients of mAb. A growing research effort to "humanize" mAb has focused primarily on the structure or sequence of the antibody variable (V) region domains. However, these approaches may ultimately suffer, as they overlook the requirement of T cell help for the immune counter-reaction and the potential of somatic hypermutation and V-D-J recombination to generate target T cell epitopes within mAb V regions. My approach focuses on this issue. In order to understand some basic principals concerning anti-immunoglobulin immune responses, I have developed a panal of T cell hybridomas and new transgenic mice. Studies with these tools strongly support our basic hypothesis that T cells are tolerant of endogenous immunoglobulin-derived diversity., Original contains color plates: All DTIC reproductions will be in black and white.

The potential of mononclonal antibodies, (mAbs), for use in therapeutic and diagnostic applications has not been fully realized in part due to counter-immune responses that often arise in patient recipients of mAb. A growing research effort to humanize mAb has focused primarily on the structure or sequence of the antibody variable (V) region domains. However, these approaches may ultimately suffer, as they overlook the requirement of T cell help for the immune counter-reaction and the potential of somatic hypermutation and V-D-J recombination to generate target T cell epitopes within mAb V regions. My approach focuses on this issue. In order to understand some basic principals concerning anti-immunoglobulin immune responses, I have developed methods, reagents and mouse strains during the previous funding year to effectively study T cell epitopes in mAb V regions. I was able to reproducibly distinguish an immune response to a V-region T cell epitope via a lymph node proliferation assay. Furthermore, I have generated MHC-tetrameric staining reagents with which I can identify T cells reactive with two such epitopes. Finally, I have generated congenic mouse strains containing or lacking the enzyme terminal deoxynucleotidyl transferase, (TdT), in order to address the role of T cell epitopes in junctional diversity.

In a previous study, CNA analysts used data from past DoD A-76 competitions to construct a model of savings and projected the potential savings from additional DoD Commercial Activities (CA) competitions. In this paper, we use an alternative approach for estimating savings from future DoD CA competitions. We estimate two separate bidding equations - one for the in-house team bid and another for the minimum contractor bids, along with an equation for baseline cost. Based on these estimated equations, one could then indirectly project future savings in the A-76 inventory as the difference between predicted baseline cost and the predicted winning bid. Using the new approach, we project an annual savings of $6 billion if the entire 1995 DoD CA inventory were competed under A-76 rules.

In 1955, the Office of Management and Budget (0MB) implemented a policy known as the Commercial Activities (CA) Program. This program enables the private sector to compete with government organizations in providing goods and services when it is appropriate and economical to do so. The objective is to promote an efficient support structure through competition. This research memorandum is part of a CNA-initiated research effort examining the DoD Commercial Activities program. Most of our previous research has examined the Navy CA program. This paper presents the results of all completed DoD comprehensive A-76 competitions between 1978 and 1994.