Andrea L. Suárez

Endothelial Cells Support Persistent Gammaherpesvirus 68 Infection

A variety of human diseases are associated with gammaherpesviruses, including neoplasms of lymphocytes (e.g. Burkitts lymphoma) and endothelial cells (e.g. Kaposis sarcoma). Gammaherpesvirus infections usually result in either a productive lytic infection, characterized by expression of all viral genes and rapid cell lysis, or latent infection, characterized by limited viral gene expression and no cell lysis. Here, we report characterization of endothelial cell infection with murine gammaherpesvirus 68 (γHV68), a virus phylogenetically related and biologically similar to the human gammaherpesviruses. Endothelial cells supported γHV68 replication in vitro, but were unique in that a significant proportion of the cells escaped lysis, proliferated, and remained viable in culture for an extended time. Upon infection, endothelial cells became non-adherent and altered in size, complexity, and cell-surface protein expression. These cells were uniformly infected and expressed the lytic transcription program based on detection of abundant viral gene transcripts, GFP fluorescence from the viral genome, and viral surface protein expression. Additionally, endothelial cells continued to produce new infectious virions as late as 30 days post-infection. The outcome of this long-term infection was promoted by the γHV68 v-cyclin, because in the absence of the v-cyclin, viability was significantly reduced following infection. Importantly, infected primary endothelial cells also demonstrated increased viability relative to infected primary fibroblasts, and this increased viability was dependent on the v-cyclin. Finally, we provide evidence for infection of endothelial cells in vivo in immune-deficient mice. The extended viability and virus production of infected endothelial cells indicated that endothelial cells provided a source of prolonged virus production and identify a cell-type specific adaptation of gammaherpesvirus replication. While infected endothelial cells would likely be cleared in a healthy individual, persistently infected endothelial cells could provide a source of continued virus replication in immune-compromised individuals, a context in which gammaherpesvirus-associated pathology frequently occurs.

Gammaherpesvirus 68 Infection of Endothelial Cells Requires both Host Autophagy Genes and Viral Oncogenes for Optimal Survival and Persistence

ABSTRACT Gammaherpesvirus-associated neoplasms include tumors of lymphocytes, epithelial cells, and endothelial cells (ECs). We previously showed that, unlike most cell types, ECs survive productive gammaherpesvirus 68 (γHV68) infection and achieve anchorage-independent growth, providing a cellular reservoir for viral persistence. Here, we demonstrated autophagy in infected ECs by analysis of LC3 localization and protein modification and that infected ECs progress through the autophagosome pathway by LC3 dual fluorescence and p62 analysis. We demonstrate that pharmacologic autophagy induction results in increased survival of infected ECs and, conversely, that autophagy inhibition results in death of infected EC survivors. Furthermore, we identified two viral oncogenes, v-cyclin and v-Bcl2, that are critical to EC survival and that modify EC proliferation and survival during infection-induced autophagy. We found that these viral oncogenes can also facilitate survival of substrate detachment in the absence of viral infection. Autophagy affords cells the opportunity to recover from stressful conditions, and consistent with this, the altered phenotype of surviving infected ECs was reversible. Finally, we demonstrated that knockdown of critical autophagy genes completely abrogated EC survival. This study reveals a viral mechanism which usurps the autophagic machinery to promote viral persistence within nonadherent ECs, with the potential for recovery of infected ECs at a distant site upon disruption of virus replication.

Viral Cyclins Mediate Separate Phases of Infection by Integrating Functions of Distinct Mammalian Cyclins

Gammaherpesvirus cyclins have expanded biochemical features relative to mammalian cyclins, and promote infection and pathogenesis including acute lung infection, viral persistence, and reactivation from latency. To define the essential features of the viral cyclin, we generated a panel of knock-in viruses expressing various viral or mammalian cyclins from the murine gammaherpesvirus 68 cyclin locus. Viral cyclins of both gammaherpesvirus 68 and Kaposis sarcoma-associated herpesvirus supported all cyclin-dependent stages of infection, indicating functional conservation. Although mammalian cyclins could not restore lung replication, they did promote viral persistence and reactivation. Strikingly, distinct and non-overlapping mammalian cyclins complemented persistence (cyclin A, E) or reactivation from latency (cyclin D3). Based on these data, unique biochemical features of viral cyclins (e.g. enhanced kinase activation) are not essential to mediate specific processes during infection. What is essential for, and unique to, the viral cyclins is the integration of the activities of several different mammalian cyclins, which allows viral cyclins to mediate multiple, discrete stages of infection. These studies also demonstrated that closely related stages of infection, that are cyclin-dependent, are in fact genetically distinct, and thus predict that cyclin requirements may be used to tailor potential therapies for virus-associated diseases.

Unusual presentation of cactus spines in the flank of an elderly man: a case report

IntroductionSplinters and spines of plant matter are common foreign bodies in skin wounds of the extremities, and often present embedded in the dermis or subcutaneous tissue. Vegetative foreign bodies are highly inflammatory and, if not completely removed, can cause infection, toxic reactions, or granuloma formation. Older patients are at increased risk for infection from untreated plant foreign bodies. The most common error in plant splinter and spine management is failure to detect their presence.Case presentationHere we report a case of cactus spines in an 84-year-old Caucasian man presenting on the right flank as multiple, red papules with spiny extensions. This presentation was unusual both in location and the spinous character of the lesions, and only after punch biopsy analysis was a diagnosis of cactus matter spines made.ConclusionsOur patient presented with an unusual case of cactus spines that required histopathology for identification. Skin lesions with neglected foreign bodies are a common cause of malpractice claims. If not removed, foreign bodies of the skin, particularly in elderly individuals, can result in inflammatory and infectious sequela. This report underscores the importance of thoroughly evaluating penetrating skin lesions for the presence of foreign bodies, such as splinters and spines.

Telling the Same Tale Twice: Déjà vu and the Shades of Grey in Self-Plagiarism

The integrity of medical and scientific publishing depends on responsible and honest practices by authors. Some of the practices that threaten the integrity of scientific publication include publishing the same paper in more than one journal (i.e. duplicate publication), presenting data from one study in multiple small papers (i.e. salami-slicing), copyright infringement, and text recycling [1]. While this subject is of vital interest to authors and investigators, as well as to journal editors and reviewers, it is also relevant to the readership of these publications. This chapter will explore and analyze examples of these practices.

Indoor Tanning Regulation, Enforcement, Taxation, and Policy

Tanning device use before the age of 35 is associated with a 75% increase in the risk for melanoma, a statistic prompting increased efforts towards limiting youth access to indoor tanning facilities. Numerous state, federal, and international restrictions focus on age restrictions, parental consent requirements, ultraviolet (UV) radiation exposure amount and frequency, warning labeling on the devices, taxation, and operator education. While commendable, these efforts are limited by non-compliance and insufficient enforcement of existing regulations.

Hiding Behind the Curtain: Anonomyous Versus Open Peer Review

For more than 50 years peer review has guided the decision-making process of editors regarding the merit of publishing scientific work. Peer reviews can be influenced by knowledge of the authors’ identities as well as anonymity of the referee. The value of the current peer review system apparently offsets areas of dissatisfaction that include: (a) reviewer bias as a result of knowing author identity, (b) unequal valuation of reviewer versus author anonymity, (c) resource expense for authors, reviewers, editors, and journals, and (d) inhibition of free communication. This chapter discusses the merits of hiding reviewer and author identity in the peer review process as well as the increasing importance of post publication review.

Outpatient Follow-up and Secondary Prevention for Melanoma Patients.

Health care providers and their patients jointly participate in melanoma prevention, surveillance, diagnosis, and treatment. This paper reviews screening and follow-up strategies for patients who have been diagnosed with melanoma, based on current available evidence, and focuses on methods to assess disease recurrence and second primary occurrence. Secondary prevention, including the roles of behavioral modification and chemoprevention are also reviewed. The role of follow-up dermatologist consultation, with focused physical examinations complemented by dermatoscopy, reflectance confocal microscopy, and/or full-body mapping is discussed. Furthermore, we address the inclusion of routine imaging and laboratory assessment as components of follow-up and monitoring of advanced stage melanoma. The role of physicians in addressing the psychosocial stresses associated with a diagnosis of melanoma is reviewed.

Ghost Busting in Dermatology Publications: Providing Byline Integrity

In this chapter we review the topic of ghostwriting and discuss the ethical discordance between commercial handling of scientific literature and research integrity. Ghostwriting puts patients at risk by providing biased information for medical practice. Multiple parties engage in and promote ghost-managed articles and practices. Efforts should be directed at enhancing awareness of ghost practices and establishing alternatives to participation in marketing in the guise of scientific research. Journal editors, reviewers, and authors should be educated on the mechanisms and detection of ghostwriting, and take appropriate measures to require financial disclosures are identified. Editors and authors have an obligation to refrain from participating in ghostwritten articles as they jeopardize public health in exchange for financial or reputational gain.