The involvement of epithelial cells in arenavirus-induced pathogenesis.

Mammalian Arenaviruses are a geographically and genetically diverse family of viruses, which is separated into two sub-groups; the Old World (OW) and New World (NW) groups. Of the OW viruses, Lassa virus (LASV), found endemically in Western Africa, is an important human pathogen, causing hundreds of thousands of infections, and several thousand deaths annually. Interestingly, some villages in endemic regions, up to 45% of the population show seropositivity for the virus. It is hypothesized that seropositivity is a result of natural infection through inhalation or ingestion of infectious particles. However, the exact mechanism is still unknown. LASV’s natural reservoir is Mastomys natalensis, a common rat found in sub-Saharan Africa. Epidemiological studies have identified the inhalation, and/or ingestion of infectious rodent excreta as the primary route of transmission from rodent reservoir to human hosts. Additionally, controlled experiments investigating intragastric (i.g.) versus intravenous (i.v.) routes of inoculation of non-human primates (NHPs) have continued transmission through these routes. viii These studies utilized Lymphocytic Choriomeningitis Virus (LCMV)-WE, a strain of LCMV that results in Lassa Fever (LF)-like disease in NHPs, and LCMVArmstrong (ARM), a strain of LCMV that mimics subclinical infection. When administered i.v., LCMV-WE-infected NHPs became systemically infected, showing clinical signs much like that of LF, and died. However, when orally infected with this virus through i.g. inoculation, some of these animals recovered, and later, were protected from lethal doses of i.v. WE challenge. Due to the nature of natural transmission from rodent to humans, epithelial cells are amongst the first cells to come in contact with the virus. However, the role(s) of the epithelial barrier during these infections have yet to be investigated. In order to investigate the role of these cells during arenaviral infection, here, a cell culture model was developed to investigate the interaction of OW mammalian arenaviruses at the site of intragastric inoculation. An important finding of this works is that the patterns of entry and release are viral dependent, and attachment to epithelial surfaces may play a role in these phenomena. Furthermore, regardless of their pathogenic potential in NHPs, both strains of LCMV, as well as LASV’s close relative, MOPV, showed similar patterns of entry and release when exposed to the apical and basolateral surfaces of polarized intestinal epithelia. Additionally, the replication patters of vaccine candidate ML- 29; a reassortant virus that contains the L segment of MOPV, and S segment of LASV, providing the exact same GP1 of LASV, were characterized. Interestingly, ML-29 virus entered and released in a different pattern than was observed with LCMV and MOPV. ix To determine if patterns of viral entry and release were driven by attachment differences, LCMV, MOPV, and ML-29 viral attachment to the surface of polarized epithelia was analyzed. LCMV an MOPV attaches to the apical and basolateral surfaces of the cells with similar efficiency. However, ML-29 showed decreased attachment to the basolateral surface of these cells as compared to the apical surface. Due to differences in pathogenicity seen in NHPs infected i.v. with LCMVWE and ARM, we hypothesized that these viruses would show differences in entry and release patterns in the polarized Caco-2 cells. However, these viruses replicated in much the same way. From these observations, we sought to further investigate differences in viral replication that may explain pathogenic differences between these closely related viruses. To do so, we investigated intracellular trafficking under the hypothesis that it may be responsible for these differences. Through the use of chemical inhibitors and immunofluorescence with confocal microscopy, a number of differences through the intracellular trafficking of LCMV-ARM and WE. The data indicates that LCMV-WE bypasses the TLR-2 receptor interaction in early endosome, and does not produce an IL-6 response in infected macrophages, opposed to LCMV-ARM-infected cells. Additionally, co-staining with LCMV and late endosome marker RAB7, showed more colocalization with LCMV-ARM than that of LCMV-WE. Furthermore, when blocking acidification of late endosome/lysosome with bafilomycin treatments, LCMV-ARM was more sensitive to pH change in the late endosome, indicating x that fusion occurs at less acidic conditions. These less acidic conditions promote earlier release at viral RNA in the case of LCMV-ARM versus that of WE. Together, these results signify differences in viral replication are tissue and viral specific. Furthermore, this research provides a platform to continue investigating key differences in viral replication between viruses of close genetic relatedness

Protecting the Diversity of the Depths: Strengthening the International Law Framework

It is only in recent decades that marine scientific research has begun to reveal the true physical characteristics and resource potential of the open ocean and deep seabed beyond national jurisdiction. A combination of factors such as the depletion of inshore fish stocks and an increase in global maritime trade has led to greater usage of the vast maritime area beyond the territorial sea and exclusive economic zone limits of the coastal states. Human activities in this area of the ocean, which covers approximately 50% of the world’s surface, have expanded to include bioprospecting, exploration for deep seabed minerals, more sophisticated marine scientific research and deep sea tourism. This rise in human activities beyond the offshore zones of coastal states poses actual and potential threats to the physical characteristics and biodiversity of the open ocean and deep sea environments. Arbitrary human intrusions into this largely unexplored marine domain have the potential to harm the intricate links between complex marine ecosystems and to erode components of marine biodiversity. This thesis examines the global and regional provisions which have been put in place to regulate the environmental impacts of human activities that occur beyond national jurisdiction. An analysis of these instruments and their implementation reveals that the current international law framework provides only minimal levels of protection for the marine environment beyond national jurisdiction. It explores several options based on the 1982 United Nations Convention on the Law of the Sea (LOSC) and the 1992 Convention on Biological Diversity (CBD) to establish a cohesive environmental protection system for the marine environment beyond national jurisdiction

The Nation\u27s Two Measures of Homicide

The United States uses two national data collection systems to track detailed information on homicides: the Federal Bureau of Investigation’s Supplementary Homicide Reports and the Centers for Disease Control and Prevention’s Fatal Injury Reports. Both measures were developed as part of a federal effort to improve national statistical systems in the early twentieth century and have gone through a number of changes since then to improve their consistency and coverage. Each program provides valuable information on the nature, trends, and patterns of homicides in the United States. Although the two measures generally capture information on the same types of events, they are designed for distinct purposes and collect different types of information. In combination, however, they produce a fairly comprehensive understanding of homicide, the most serious form of violence

Building a Sustainable Comprehensive Women's Health Program: The Michigan Model

The Women's Health Program at the University of Michigan was established in 1993 and has developed into a successful, federally supported program that links clinical research and education activities across the University. It has focused on human resource capacity building, sustainable financial support and infrastructure, and adaptability to change and opportunities. Widely accepted standards, demonstrated value, committed leaders/champions, and participatory culture have contributed to its success and are important to its future.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63102/1/jwh.2006.0192.pd

Bordetella petrii Clinical Isolate

We describe the first clinical isolate of Bordetella petrii from a patient with mandibular osteomyelitis. The only previously documented isolation of B. petrii occurred after the initial culture of a single strain from an environmental source

Effect of the human papillomavirus (HPV) quadrivalent vaccine in a subgroup of women with cervical and vulvar disease: retrospective pooled analysis of trial data

Objectives To determine the effect of human papillomavirus (HPV) quadrivalent vaccine on the risk of developing subsequent disease after an excisional procedure for cervical intraepithelial neoplasia or diagnosis of genital warts, vulvar intraepithelial neoplasia, or vaginal intraepithelial neoplasia