Toll-like Receptors 2 and 9 Are Important for Innate Immune Cell Activation and Recruitment in Hypersensitivity Pneumonitis

Hypersensitivity pneumonitis (HP) is an interstitial lung disease caused by repeated inhalation of environmental antigens. The disease is characterized by alveolitis and granuloma formation; however, some patients develop chronic HP (CHP), a restrictive lung disease characterized by fibrosis. Previous studies revealed that neutrophils are recruited into the lung via a MyD88- dependent pathway and regulate disease severity through cytokine production. Toll-like receptors (TLRs) 2 and 9 recognize conserved molecular patterns present in bacteria, and signal through the adaptor molecule MyD88. The goal of my project is to investigate the role of TLRs 2 and 9 in the pathogenesis of HP during the acute phase, granulomatous phase, and chronic phase of the disease. Using the S. rectivirgula (SR) animal model of HP, our studies indicated that individually, TLRs 2 and 9 contributed to neutrophil recruitment. We generated TLR2/9 double knockout (TLR2/9−/− ) mice to determine the extent to which TLRs 2 and 9 cooperate in neutrophil recruitment during HP. During the acute response, TLR2/9−/− mice exposed to SR demonstrated a significant reduction in neutrophil recruitment after a single exposure to SR compared to C57BL/6 mice (WT). The reduction in neutrophils was associated with a reduction in the expression of the neutrophil chemokine CXCL2 and inflammatory cytokines TNFα and IL-6 in the BALF. Disease severity in HP is associated with granuloma formation and a Th17 response. After 3 weeks exposure to SR, our results demonstrate a decrease in IL-17 and IL-22 mRNA expression and a corresponding decrease in the percentage of Th17 cells in TLR2/9−/− mice compared to WT and single knockout (SKO) mice. The decrease in Th17 cells in TLR2/9−/− mice was not associated with a significant increase in T regulatory (Treg) cells or a switch to a Th1 response. In addition, TLR2−/− and TLR2/9−/− mice have significantly fewer activated CD4+ Th cells. Interstitial macrophages from TLR2/9−/− mice have decreased costimulatory molecule and MHCII expression suggesting they are deficient in T cell activation. However, TLR2−/− , TLR9−/− , and TLR2/9−/− still form granulomas in the lung. To determine the extent to which TLRs 2 and 9 contributed to the development of CHP, WT and TLR2/9−/− mice were intranasally exposed to SR three times/week for 15 weeks. WT and TLR2/9−/− mice developed a neutrophilic and lymphocytic alveolitis, but TLR2/9−/− mice had a significant increase in eosinophils in the bronchoalveolar lavage (BAL) fluid compared to WT exposed mice. Th17 cells, but not Th2 cells were detected in the lungs of mice exposed to SR for 14 weeks, suggesting CHP is not associated with a switch to a Th2 type immune response. WT and TLR2/9−/− mice had significantly reduced static compliance compared to WT unexposed mice. To determine whether the restrictive lung defect was associated with fibrosis, we visualized collagen in the lung by histology using Masson’s trichrome stain. Measuring collagen staining by positive pixel analysis suggested TLR2/9−/− mice were partially protected from lung fibrosis compared to WT mice. Altogether, the results suggest TLRs 2 and 9 cooperate in neutrophil recruitment and subsequently disease severity and play an integral role in the outcome of HP

Visualising apoptosis in live zebrafish using fluorescence lifetime imaging with optical projection tomography to map FRET biosensor activity in space and time

Fluorescence lifetime imaging (FLIM) combined with optical projection tomography (OPT) has the potential to map Förster resonant energy transfer (FRET) readouts in space and time in intact transparent or near transparent live organisms such as zebrafish larvae, thereby providing a means to visualise cell signalling processes in their physiological context. Here the first application of FLIM OPT to read out biological function in live transgenic zebrafish larvae using a genetically expressed FRET biosensor is reported. Apoptosis, or programmed cell death, is mapped in 3-D by imaging the activity of a FRET biosensor that is cleaved by Caspase 3, which is a key effector of apoptosis. Although apoptosis is a naturally occurring process during development, it can also be triggered in a variety of ways, including through gamma irradiation. FLIM OPT is shown here to enable apoptosis to be monitored over time, in live zebrafish larvae via changes in Caspase 3 activation following gamma irradiation at 24 hours post fertilisation. Significant apoptosis was observed at 3.5 hours post irradiation, predominantly in the head region

Application of advanced technologies to small, short-haul aircraft

The results of a preliminary design study which investigates the use of selected advanced technologies to achieve low cost design for small (50-passenger), short haul (50 to 1000 mile) transports are reported. The largest single item in the cost of manufacturing an airplane of this type is labor. A careful examination of advanced technology to airframe structure was performed since one of the most labor-intensive parts of the airplane is structures. Also, preliminary investigation of advanced aerodynamics flight controls, ride control and gust load alleviation systems, aircraft systems and turbo-prop propulsion systems was performed. The most beneficial advanced technology examined was bonded aluminum primary structure. The use of this structure in large wing panels and body sections resulted in a greatly reduced number of parts and fasteners and therefore, labor hours. The resultant cost of assembled airplane structure was reduced by 40% and the total airplane manufacturing cost by 16% - a major cost reduction. With further development, test verification and optimization appreciable weight saving is also achievable. Other advanced technology items which showed significant gains are as follows: (1) advanced turboprop-reduced block fuel by 15.30% depending on range; (2) configuration revisions (vee-tail)-empennage cost reduction of 25%; (3) leading-edge flap addition-weight reduction of 2500 pounds

Advance telephone calls ahead of reminder questionnaires increase response rate in non-responders compared to questionnaire reminders only : The RECORD phone trial

Peer reviewedPublisher PD

Successes, challenges and lessons learned: Community-engaged research with South Carolina's Gullah population

Engaging communities is highly recommended in the conduct of health research among vulnerable populations. The strength of community-engaged research is well documented and is recognised as a useful approach for eliminating health disparities and improving health equity. In this article, five interdisciplinary teams from the Medical University of South Carolina present their involvement with community-engaged research with a unique population of Gullah African Americans residing in rural South Carolina. Their work has been integrated with the nine established principles of community-engaged research: establishing clear goals, becoming knowledgeable about the community, establishing relationships, developing community self-determination, partnering with the community, maintaining respect, mobilising community assets, releasing control, and maintaining community collaboration. In partnership with a Citizen Advisory Committee, developed at the inception of the first community-engaged research project, the academic researchers have been able to build on relationships and trust with this population to sustain partnerships and to meet major research objectives over a 20-year period. Challenges observed include structural inequality, organisational and cultural issues, and lack of resources for building sustainable research infrastructure. Lessons learned during this process include the necessity for clearly articulated and shared goals, knowledge about the community culture, and embedding the cultural context within research approaches. Keywords: Engaged health research, vulnerable populations, longterm collaboration, South Carolina 'Gullah' communitie

Genetic and epigenetic regulation in Lingo-1 : Effects on cognitive function and white matter microstructure in a case-control study for schizophrenia

Leucine-rich repeat and immunoglobulin domain-containing protein (Lingo-1) plays a vital role in a large number of neuronal processes underlying learning and memory, which are known to be disrupted in schizophrenia. However, Lingo-1 has never been examined in the context of schizophrenia. The genetic association of a single-nucleotide polymorphism (SNP, rs3144) and methylation (CpG sites) in the Lingo-1 3′-UTR region was examined, with the testing of cognitive dysfunction and white matter (WM) integrity in a schizophrenia case-control cohort (n = 268/group). A large subset of subjects (97 control and 161 schizophrenia subjects) underwent structural magnetic resonance imaging (MRI) brain scans to assess WM integrity. Frequency of the rs3144 minor allele was overrepresented in the schizophrenia population (p = 0.03), with an odds ratio of 1.39 (95% CI 1.016–1.901). CpG sites surrounding rs3144 were hypermethylated in the control population (p = 0.032) compared to the schizophrenia group. rs3144 genotype was predictive of membership to a subclass of schizophrenia subjects with generalized cognitive deficits (p < 0.05), in addition to having associations with WM integrity (p = 0.018). This is the first study reporting a potential implication of genetic and epigenetic risk factors in Lingo-1 in schizophrenia. Both of these genetic and epigenetic alterations may also have associations with cognitive dysfunction and WM integrity in the context of the schizophrenia pathophysiology

Roles of residues 3 and 4 in cyclic tetrapeptide ligand recognition by the Κ -opioid receptor

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/75723/1/j.1399-3011.2005.00220.x.pd

Starting School: a large-scale start of school assessment within the ‘Born in Bradford’ longitudinal cohort [version 1; peer review: 1 approved with reservations]

The Born in Bradford (BiB) cohort of 13,776 children born between 2007-2011 and their parents provides a rich data resource for researchers exploring protective and risk factors influencing long-term developmental and health outcomes. Educational attainment is a critical factor related to later health. Literacy and communication, fine motor skills and social and emotional health are key ‘early’ predictors of educational attainment and can be used to identify children in need of additional support. We describe our BiB ‘Starting School’ data collection protocol which assessed literacy and communication, fine motor skills and social and emotional health on 3,444 BiB children aged 4-5 years old. These measures supplement the existing dataset, and complement the routine educational, health and social care data available for the cohort

Comparing the Outcomes of Face-to-Face and Synchronous Online Research Mentor Training Using Propensity Score Matching

In this study, propensity score matching (PSM) was conducted to examine differences in the effectiveness of research mentor training (RMT) implemented using two modes-face-to-face or synchronous online training. This study investigated each training mode and assessed participants\u27 perceived gains in mentoring skills, ability to meet mentees\u27 expectations, and overall quality of mentoring, as well as intention to make changes to their mentoring practices. Additional factors that may contribute to participant outcomes were also examined. In total, 152 mentors trained using a synchronous online platform and 655 mentors trained in in-person workshops were analyzed using the PSM method. Mentors were matched based on similar characteristics, including mentee\u27s career stage, mentor\u27s title, mentor\u27s prior mentoring experience, mentor\u27s race/ethnicity and sex, and mentor\u27s years of experience; results show that both face-to-face and synchronous online modes of RMT are effective. Findings indicated that the training mode did not significantly impact the mentors\u27 perceived training outcomes. Factors associated with the reported training outcomes included dosage (hours of training), facilitator effectiveness, race/ethnicity, and previous mentoring experience. The results of this study demonstrate that mentors\u27 perceived training outcomes are comparable regardless of the training modality used-online versus face-to-face

Body Composition and Blood Biomarkers Correlate with Muscle Strength and Anaerobic Performance

Blood biomarkers and body composition are sensitive indicators to monitor and predict performance in the athletic population. The analysis of these markers can help monitor progression in athletes competing in aerobic and anaerobic sports; however, it is unclear which biomarkers are associated with muscle strength and anaerobic performance. PURPOSE: To explore the correlations of body composition and blood biomarkers with muscle strength and power tests of the countermovement jump (CMJ), broad jump (BJ), isometric mid-thigh pulls (IMTP), hand grip strength (HGS), and the Wingate anaerobic test. METHODS: This study recruited nine collegiate intramural sports club students (Height=171.43±10.05 cm, Weight=70.36±13.18 kg, Age=23.87±3.65 years). Body composition was assessed with air displacement plethysmography (BodPod), and muscular strength was measured by IMTP and HGS. Muscular power was assessed via CMJ and BJ and calculated from ground reaction forces. Additionally, peak power output and mean power output were determined from the 30-second Wingate test. Blood biomarkers of hemoglobin (Hb), red blood cell count (RBC), and hematocrit (Hct) were analyzed by Quest Diagnostics. Data normality was confirmed with a Shapiro-Wilk test. Pearson-R correlations were conducted to determine associations between body composition, blood biomarkers, and the above performance tests. Significance was set at an alpha level of p\u3c0.05. RESULTS: Fat free mass was positively correlated with BJ distance (r=0.805, p=0.016), IMTP peak force (r=0.907, p=0.002), Wingate peak power (r=0.708, p=0.049), and Wingate mean power (r=0.910, p=0.002). Additionally, body fat percentage was negatively associated with IMTP peak force (r=-0.850, p=0.008) and mean power in the Wingate (r=-0.800, p=0.017). There was a positive correlation between blood biomarkers (RBC, Hb, and Hct) and mean power of the Wingate (r=0.693-0.906, p\u3c0.05), and peak force of the IMTP (r=0.710–0.833, p\u3c0.05). Furthermore, RBC was found positively correlated with CMJ height (r=0.815, p=0.007) and BJ distance (r=0.763, p=0.028), while HGS was positively associated with RBC (r=0.910, p=0.012) and Hct (r=0.962, p=0.002). CONCLUSION: Body composition and blood biomarkers correlate with muscle strength, power, and anaerobic performance, which could be potentially used to monitor and predict human performance in athletes