Longitudinal histopathological, immunohistochemical, and In Situ hybridization analysis of host and viral biomarkers in liver tissue sections of Ebola (EBOV) infected rhesus macaques

INTRODUCTION: Ebola virus (EBOV) is a highly infectious and often lethal filovirus that causes hemorrhagic fever, with a reported case fatality rate of 40-90%. There are currently no Food and Drug Administration (FDA) approved medical countermeasures (MCMs) for EBOV. Non-human primates (NHPs) remain the gold standard animal model for EBOV research as they most accurately recapitulate human disease. OBJECTIVE: This study aimed to characterize the temporal viral pathogenesis of EBOV in the liver of infected rhesus macaques using routine histopathology, multiplex immunohistochemistry (mIHC) and multiplex fluorescent In Situ Hybridization (mFISH), refined by digital pathology (DP) and image analysis (DIA). METHODS: 21 FFPE liver sections from EBOV-infected rhesus macaques were examined microscopically (Uninfected controls n=3; 3 DPE n=3; 4 DPE n=3; 5 DPE n=3; 6 DPE n=3; Terminal n=6). Tissues were stained with H&E and PTAH for histopathological scoring. Three serial sections were fluorescently immunolabeled or hybridized under three independent conditions (1.EBOV VP35, Tissue Factor, CD68; 2.EBOV VP35, Heppar, Myeloperoxidase (MPO); 3.EBOV VP35, IL-6, ISG-15). Slides were digitized by a Vectra PolarisTM fluorescent whole slide scanner and DIA was conducted using HaloTM image analysis software. Statistical analysis was conducted using GraphPad PrismTM 8.0. RESULTS: Comparing peracute (3-4 DPE) to acute (5-6 DPE) and terminal (6-8 DPE) EBOV infection, there is a statistically significant (p < 0.05) increase in hepatic inflammation and fibrin thrombi, correlating with an absolute increase in macrophages (CD68), neutrophils (MPO), and total % of Tissue Factor in the liver. There is also a significant increase in the severity of necrosis, which correlates with a decrease in Heppar. While there was significant colocalization of VP35 and CD68 starting at 4 DPE, there was only rare colocalization of VP35 with Heppar, even in terminal animals. Similar to mIHC, progressive and statistically significant differences were observed in gene expression when comparing peracute to acute and terminal EBOV infection. IL-6 predominated within periportal fibrovascular compartments, but also colocalized within cells concurrently expressing EBOV VP35. EBOV VP35 expression was observed within histiocytes, endothelial cells, and less commonly hepatocytes. ISG-15 expression was observed in periportal regions and in proximity to cells expressing EBOV VP35, but colocalization within EBOV VP35 expressing cells was an extremely rare event. CONCLUSION: Qualitative tools are well suited for confirming virulence and viral tissue tropism, but do little to build on our current understanding of disease. Using DIA in partnership with mIHC and mFISH, this study quantified statistically significant temporal changes in the immunoreactivity and hybridization of host and viral biomarkers that have previously been linked to the pathogenesis of EBOV. Taken together, these tools have enabled us to characterize minute changes that reflect magnitudes of biological variability simply not feasible to detect with the human eye. Furthermore, spatial context has refined our current understanding of differential gene expression of EBOV, which has the potential to aid in development of host-directed therapies. The establishment of these benchmarks will serve as a guide for the validation of cross-institutional EBOV animal models

Cobble substrate in a surface bypass reduces bypass acceptance by common roach Rutilus rutilus

Historically, ecological engineered solutions for fish passage across anthropogenic barriers in rivers has mainly focused on facilitating upstream passage for long-migrating diadromous fish, such as salmonids. More recently, passage solutions have shifted their focus to a more holistic ecological perspective, allowing passage for species with different swimming capacity, both upstream and downstream. This experiment investigated whether the addition of cobble in the passageway of a surface bypass could facilitate downstream movement of a cyprinid fish, the common roach Rutilus rutilus. Surface bypasses were constructed in large experimental flumes and roach were released into the flumes and monitored for bypass passage using PIT-telemetry through 11-h night-trials. Behavior was scored using four continuously-recording video cameras at the bypass construction. There was a negative effect of substrate-treatment on the passage rate through the bypass. The majority of the fish in the No substrate treatment had successfully passed within 4 h, while a lesser proportion of the fish in the Substrate treatment had done so (additional fish in the latter treatment passed later in the trials). Fish exposed to cobble substrate in the bypass passageway showed more avoidance-like behaviors at the ramp section of the bypass and tended to return back upstream more often than the fish in the no-substrate control trials. When reaching the passageway, the substrate-exposed fish expressed no behaviors that could be indicative of reduced passage success, as compared to controls. We conclude that passage was not hindered by the presence of cobble substrate, but passage was delayed due to avoidance behavior at the bypass ramp when cobble substrate was present. Based on these results, the addition of cobble substrate in a surface bypass cannot be recommended as a measure to facilitate the downstream passage performance of the common roach through surface bypasses

Fish biodiversity in different types of tributary mouths located within impounded sections of Swedish boreal rivers

Large boreal rivers in Sweden are generally impounded by hydropower dams and a large proportion of main stem shallow flowing habitats have been lost. Tributaries often contain the last undisturbed habitats and could be important for the conservation of species diversity. In particular, tributary mouth areas could be biodiversity hot-spots, due to their vicinity to the main stem and favorable environmental conditions. In this study, we investigate whether tributary mouth areas in two impounded boreal rivers (Ume River and Lule River) could be regarded as biodiversity hot-spots for fish. Based on standardized electrofishing in 20 tributary mouths, we find that overall fish diversity is generally low. The highest species richness and diversity was found in mouth areas dominated by intermediate substrate sizes (gravel – cobble). Few, if any, species were found in areas where fine sediments (smaller than sand) dominated. The tributary mouth areas had similar species richness and diversity as areas in the tributaries located 1-km upstream of the mouth, but the fish community composition often differed between these two types of sites. Management action favoring fish diversity in the tributary mouth areas could include protection or rehabilitation of areas dominated by medium sized substrate and reduction of erosion and transport of fine sediments in the tributaries. Overall, we find no support for tributary mouths being hot-spots for fish biodiversity and while some patterns in diversity gives hints on suitable management action, it is important to further understand impacts in tributaries and their mouths and the temporal dynamics of the fish community. Aggradation Boreal rivers Fish biodiversity River morphology River sediment size Tributary confluencepublishedVersio

The Role of Social Identity in Viewer Attitudes and Intentions

The research project aims to identify a relationship between self-identification based on the social identity theory and feelings towards a particular show. Focusing the research on four fictitious shows with similar plot lines, but different casts of the same ethnic group, the research team has created a questionnaire that asked questions relating to the shows and participant social identity. The survey sample consists of 241 individuals between the ages of 18 and 28. The results show that there is a positive predictive relationship between feeling that your social group is represented in a show and attitudes towards that show, as well as intentions to watch the show. Understanding the participant’s viewing attitudes and behaviors relating to their self-identification will provide insight to marketers that will aid in future development of media including television series and film that is more understanding of the new nuances that dictate viewership behavior

Parents' perceived obstacles to pediatric clinical trial participation: Findings from the clinical trials transformation initiative.

Enrollment of children into pediatric clinical trials remains challenging. More effective strategies to improve recruitment of children into trials are needed. This study used in-depth qualitative interviews with parents who were approached to enroll their children in a clinical trial in order to gain an understanding of the barriers to pediatric clinical trial participation. Twenty-four parents whose children had been offered the opportunity to participate in a clinical trial were interviewed: 19 whose children had participated in at least 1 clinical trial and 5 who had declined participation in any trial. Each study aspect, from the initial explanation of the study to the end of the study, can affect the willingness of parents to consent to the proposed study and future studies. Establishing trust, appropriate timing, a transparent discussion of risks and benefits oriented to the layperson, and providing motivation for children to participate were key factors that impacted parents' decisions. In order for clinical trial accrual to be successful, parents' priorities and considerations must be a central focus, beginning with initial trial design. The recommendations from the parents who participated in this study can be used to support budget allocations that ensure adequate training of study staff and improved staffing on nights and weekends. Studies of parent responses in outpatient settings and additional inpatient settings will provide valuable information on the consent process from the child's and parent's perspectives. Further studies are needed to explore whether implementation of such strategies will result in improved recruitment for pediatric clinical trials

Patient and renal allograft outcomes of anticoagulation therapy in kidney transplantation

Introduction: Kidney transplant recipients are often on anticoagulation or antiplatelet therapy, for a variety of indications. These medications are associated with increased risk of bleeding but may reduce risk of thrombosis and possible complications such as allograft loss. The aim of our study is to evaluate the effect of perioperative anticoagulation or antiplatelet therapy on patient and allograft outcomes. Methods: Data collection was done by retrospective chart review of all patients who underwent a kidney or kidney/pancreas transplantation at Thomas Jefferson University Hospital (TJUH) from 2012-2019. We recorded perioperative anticoagulation and antiplatelet therapy, risk factors for bleeding and thrombosis, post-transplant episodes of bleeding or thrombosis, allograft function at multiple timepoints from 30 days to 3 years post-transplant, allograft loss, and mortality. Data analysis will evaluate relative outcomes based on perioperative anticoagulation or antiplatelet status. Results: We have completed collecting data for the 872 kidney or kidney/pancreas transplants done at TJUH from 2012-2019 and are currently working on analyzing the data. The patients will be evaluated in groups based on the presence and type of anticoagulation or antiplatelet therapy. These groups will then be compared based on patient and allograft outcomes. Discussion: We expect to find that patients on perioperative anticoagulation or antiplatelet therapy had higher rates and severity of bleeding, but lower rates of thrombosis and therefore allograft loss and mortality. This study will improve our understanding of perioperative anticoagulation and antiplatelet therapy so we may develop treatment guidelines for the TJUH kidney transplant program

Perceived barriers to pediatrician and family practitioner participation in pediatric clinical trials: Findings from the Clinical Trials Transformation Initiative.

Despite legislation to stimulate pediatric drug development through clinical trials, enrolling children in trials continues to be challenging. Non-investigator (those who have never served as a clinical trial investigator) providers are essential to recruitment of pediatric patients, but little is known regarding the specific barriers that limit pediatric providers from participating in and referring their patients to clinical trials. We conducted an online survey of pediatric providers from a wide variety of practice types across the United States to evaluate their attitudes and awareness of pediatric clinical trials. Using a 4-point Likert scale, providers described their perception of potential barriers to their practice serving as a site for pediatric clinical trials. Of the 136 providers surveyed, 52/136 (38%) had previously referred a pediatric patient to a trial, and only 17/136 (12%) had ever been an investigator for a pediatric trial. Lack of awareness of existing pediatric trials was a major barrier to patient referral by providers, in addition to consideration of trial risks, distance to the site, and time needed to discuss trial participation with parents. Overall, providers perceived greater challenges related to parental concerns and parent or child logistical barriers than study implementation and ethics or regulatory barriers as barriers to their practice serving as a trial site. Providers who had previously been an investigator for a pediatric trial were less likely to be concerned with potential barriers than non-investigators. Understanding the barriers that limit pediatric providers from collaboration or inhibit their participation is key to designing effective interventions to optimize pediatric trial participation