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Studies have shown that infant temperament varies with maternal psychosocial factors, in utero illness, and environmental stressors. We predicted that the pandemic would shape infant temperament through maternal SARS-CoV-2 infection during pregnancy and/or maternal postnatal stress. To test this, we examined associations among infant temperament, maternal prenatal SARS-CoV-2 infection, maternal postnatal stress, and postnatal COVID-related life disruptions.We tested 63 mother-infant dyads with prenatal maternal SARS-CoV-2 infections and a comparable group of 110 dyads without infections. To assess postnatal maternal stress, mothers completed the Perceived Stress Scale 4 months postpartum and an evaluation of COVID-related stress and life disruptions 6 months postpartum. Mothers reported on infant temperament when infants were 6-months-old using the Infant Behavior Questionnaire-Revised (IBQ-R) Very Short Form.Maternal SARS-CoV-2 infection during pregnancy was not associated with infant temperament or maternal postnatal stress. Mothers with higher self-reported postnatal stress rated their infants lower on the Positive Affectivity/Surgency and Orienting/Regulation IBQ-R subscales. Mothers who reported greater COVID-related life disruptions rated their infants higher on the Negative Emotionality IBQ-R subscale.Despite no effect of prenatal maternal SARS-CoV-2 infection, stress and life disruptions incurred by the COVID-19 pandemic were associated with infant temperament at 6-months.SARS-CoV-2 infection during pregnancy is not associated with postnatal ratings of COVID-related life disruptions, maternal stress, or infant temperament. Postnatal ratings of maternal stress during the COVID-19 pandemic are associated with normative variation in maternal report of infant temperament at 6 months of age. Higher postnatal ratings of maternal stress are associated with lower scores on infant Positive Affectivity/Surgency and Orienting/Regulation at 6 months of age. Higher postnatal ratings of COVID-related life disruptions are associated with higher scores on infant Negative Emotionality at 6 months of age.

Rotavirus replication takes place in the viroplasms, which are cytosolic inclusions allowing the synthesis of virus genome segments, followed by their encapsidation in the core shell and the addition of the second layer of the virion. The viroplasms are composed of several viral proteins, including NSP5, which is the main building block. Microtubules, lipid droplets, and miRNA-7 are among the host components recruited in viroplasms. Aiming to investigate the relationship between rotavirus proteins and host components in the viroplasms, we performed a pull-down assay of lysates from rotavirus-infected cells expressing NSP5-BiolD2. Subsequent tandem mass spectrometry identified all eight subunits of the TRiC complex, a cellular chaperonin responsible for folding at least 10% of the cytosolic proteins. Our validated results show that TriC is recruited in viroplasms and there specifically surrounds newly formed double-layered particles (DLPs). Chemical inhibition of TRiC and silencing of its subunits drastically reduced virus progeny production.Interestingly, TRiC-inhibited RV-infected cells lacked TLPs but harbored empty DLPs. Using sequence-specific direct RNA nanopore sequencing, we demonstrate that TRiC is essential for RV replication since it modulates dsRNA genome segment synthesis, specifically (−)RNA. Moreover, TRiC is associated with RNA-dependent RNA polymerase, VP1, and regulates the folding of VP2, a cofactor allowing dsRNA synthesis. This study providesin vivoevidence of the regulatory mechanism by which dsRNA genome segment replication is controlled and coordinated in the rotavirus viroplasms.ImportanceThe replication of rotavirus takes place in cytosolic inclusions termed viroplasms. In these inclusions, the eleven double-stranded RNA genome segments are synthesized and packaged individually into the newly generated virus particles. In this study, we show for the first time that the TRiC complex, a cellular chaperonin responsible for the folding of at least 10% of the cytosolic proteins, is a component of viroplasms and is required for the synthesis of the viral (−)ssRNA. Specifically, TRiC interacts with VP1, the RNA-dependent RNA polymerase, and assists in the folding of VP2, the cofactor involved in RNA replication. Our study adds a new component to the current model of rotavirus replication, where TRiC is recruited in viroplasm to assist replication.

Neuroimaging studies point to neurostructural abnormalities in youth with anxiety disorders. Yet, findings are based on small-scale studies, often with small effect sizes, and have limited generalizability and clinical relevance. These issues have prompted a paradigm shift in the field towards highly powered (i.e., big data) individual-level inferences, which are data-driven, transdiagnostic, and neurobiologically informed. Here, we built and validated neurostructural machine learning (ML) models for individual-level inferences based on the largest-ever multi-site neuroimaging sample of youth with anxiety disorders (age: 10-25 years, N=3,343 individuals from 32 global sites), as compiled by three ENIGMA Anxiety Working Groups: Panic Disorder (PD), Generalized Anxiety Disorder (GAD), and Social Anxiety Disorder (SAD). ML classifiers were trained on MRI-derived regional measures of cortical thickness, surface area, and subcortical volumes to classify patients and healthy controls (HC) for each anxiety disorder separately and across disorders (transdiagnostic classification). Modest, yet robust, classification performance was achieved for PD vs. HC (AUC=0.62), but other disorder-specific and transdiagnostic classifications were not significantly different from chance. However, above chance-level transdiagnostic classifications were obtained in exploratory subgroup analyses of male patients vs. male HC, unmedicated patients vs. HC, and patients with low anxiety severity vs. HC (AUC 0.59-0.63). The above chance-level classifications were based on plausible and specific neuroanatomical features in fronto-striato-limbic and temporo-parietal regions. This study provides a realistic estimate of classification performance in a large, ecologically valid, multi-site sample of youth with anxiety disorders, and may as such serve as a benchmark.

Human papillomavirus (HPV)-induced carcinogenesis is associated with unregulated expression of the oncoproteins E6 and E7. HPV E7 is a viral protein that lacks enzymatic activity; however, it can target several cellular proteins to induce cell transformation and promote uncontrolled proliferation. Although several E7 targets have been described, there are still gaps in the understanding of how this oncoprotein drives cells toward malignancy. Here, using a small HPV type 16 (HPV16) E7 peptide in a proteomic approach, we report Memo1 as a new E7 binding partner, interacting through the aspartic and glutamic acid residues (E80 and D81) in the C-terminal region of HPV16 E7. Furthermore, we demonstrate that HPV16 E7 targets Memo1 for proteasomal degradation through a Cullin2-dependent mechanism. In addition, we show that overexpression of Memo1 decreases cell transformation and proliferation and that reduction of Memo1 levels correlate with activation of Akt and an increase in invasion of HPV-positive cervical cancer cell lines. Our results show a novel HPV E7 interacting partner and describe novel functions of Memo1 in the context of HPV-induced malignancy. IMPORTANCE Although numerous targets have been reported to interact with the HPV E7 oncoprotein, the mechanisms involved in HPV-induced carcinogenesis and the maintenance of cell transformation are still lacking. Here, through pulldown assays using a peptide encompassing the C-terminal region of HPV16 E7, we report Memo1 as a novel E7 interactor. High levels of Memo1 correlated with reduced cell proliferation and, concordantly, knockdown of Memo1 resulted in Akt activation in HPV-positive cell lines. These results highlight new mechanisms used by HPV oncoproteins to modulate proliferation pathways in cervical cancer cells and increase our understanding of the link between Memo1 protein and cancer.

Combinations of dielectric and NMR measurements are frequently used to improve saturation modeling in complex situations, often incorporating the concept of wettability. Due to the two methods' distinct tools and physical mechanisms, the interplay of the electrical and magnetic fields and their constitutive equations are generally not addressed. This is directly counter to the situation with the medical imaging modalities, magnetic resonance electrical properties tomography (MREPT) and magnetic resonance electrical impedance tomography (MREIT), where field-specific polarizations and relaxations are used to enhance the contrast. Both electrical and magnetic (EM) fields at the frequencies typically encountered in laboratory and logging environments impart molecular motions impacted by pore structure. In both instances, restricted motions are reflected in their individual responses' time or frequency domain. Using time-domain relaxations and variations in both EM fields, this work focuses on the practicality of using NMR and dielectric relaxation comparisons originally proposed by Bloembergen, Purcell, and Pound (BPP). Similar to the dipolar relaxation equivalence in the BPP model, we develop a relaxation time correlation assuming representative Maxwell-Wagner relaxations for the key pore components demonstrated by Myers. The distributions of dielectric relaxation times evident in carbonate dispersion curves from 1 – 300 MHz were quantified using the Havriliak-Negami (HN) model. The quantifications are then used to evaluate characteristic dielectric dispersions curves generated from a dielectric model introducing multiple pore systems in carbonates. The modeled distributions are spectrally mapped to the NMR T2 distributions based on Debye shielding distances correlated with the conductivity. The interplay of pore connectivity and surface and bulk diffusivity are modeled using a "two-fraction fast exchange model" by Brownstein and Tarr. Using dielectric and NMR experiments along with a combination of micro-CT and SEM imaging techniques, the NMR-based spectral distribution of dielectric relaxation times demonstrates that variable-length scales and fractal dimensions accessed through the dielectric dispersion measurements are more extensive than that implied by the standard reference to the "texture" of a carbonate sample. We also show that the modeled distributions are closely correlated with the conductivity and provide improved petrophysical insight for the frequently used Archie exponent combination (MN) associated with the water tortuosity.

HIGHLIGHTS • Nutrition-based interventions to reduce COVID-19 morbidity/mortality are needed. • The Omega-3 Index (O3I) was measured in banked blood from 100 COVID-19 patients. • Risk for death was analyzed as a function of quartiles (Q) of the O3I. • Patients in Q4 (O3I ≥ 5.7%) vs. Q1-3 were 75% less likely to die (p=0.07). • These pilot data suggest that a higher O3I may lower risk for death from COVID-19., Very-long chain omega-3 fatty acids (EPA and DHA) have anti-inflammatory properties that may help reduce morbidity and mortality from COVID-19 infection. We conducted a pilot study in 100 patients to test the hypothesis that RBC EPA+DHA levels (the Omega-3 Index, O3I) would be inversely associated with risk for death by analyzing the O3I in banked blood samples drawn at hospital admission. Fourteen patients died, one of 25 in quartile 4 (Q4) (O3I ≥5.7%) and 13 of 75 in Q1-3. After adjusting for age and sex, the odds ratio for death in patients with an O3I in Q4 vs Q1-3 was 0.25, p=0.07. Although not meeting the classical criteria for statistical significance, this strong trend suggests that a relationship may indeed exist, but more well-powered studies are clearly needed.

Precise identification of drinking and smoking patterns during pregnancy is crucial to better understand the risk to the fetus. The purpose of this manuscript is to describe the methodological approach used to define prenatal drinking and smoking trajectories from a large prospective pregnancy cohort, and to describe maternal characteristics associated with different exposure patterns. In the Safe Passage Study, detailed information regarding quantity, frequency, and timing of exposure was self-reported up to four times during pregnancy and at 1 month post-delivery. Exposure trajectories were developed using data from 11,692 pregnancies (9,912 women) where pregnancy outcome was known. Women were from three diverse populations: white (23%) and American Indian (17%) in the Northern Plains, US, and mixed ancestry (59%) in South Africa (other/not specified [1%]). Group-based trajectory modeling was used to identify 5 unique drinking trajectories (1 none/minimal, 2 quitting groups, 2 continuous groups) and 7 smoking trajectories (1 none/minimal, 2 quitting groups, 4 continuous groups). Women with pregnancies assigned to the low- or high-continuous drinking groups were less likely to have completed high school and were more likely to have enrolled in the study in the third trimester, be of mixed ancestry, or be depressed than those assigned to the none/minimal or quit-drinking groups. Results were similar when comparing continuous smokers to none/minimal and quit-smoking groups. Further, women classified as high- or low-continuous drinkers were more likely to smoke at moderate-, high-, and very high-continuous levels, as compared to women classified as non-drinkers and quitters. This is the first study of this size to utilize group-based trajectory modeling to identify unique prenatal drinking and smoking trajectories. These trajectories will be used in future analyses to determine which specific exposure patterns subsequently manifest as poor peri- and postnatal outcomes.

Advances in ICT have changed, and continue to change, interactions between service providers and customers. Service industries like health care or consulting traditionally relied on interpersonal “high touch, low tech” (Bitner, Brown, and Meuter 2000: 138) exchanges. Today, however, service providers and customers increasingly interact through virtual, rather than physical interfaces (Breidbach, Kolb, and Srinivasan 2013a). But, service research to date has focused predominantly on face-to-face settings (e.g., Froehle and Roth 2004), while technology-enabled value co-creation processes remain largely unexplored and misunderstood (Breidbach and Maglio 2015). Consequently, the understanding of ICT-enabled service is incomplete, and exploring the broader role and implications of ICT in service represents a key research priority for service science (e.g., Srinivasan, Breidbach, and Kolb 2015) and IS scholars alike (Maglio and Breidbach 2014).

The focus of the health sector in most countries remains almost exclusively on health care services, and the potential of multisectoral collaboration remains untapped in many low- and middle-income countries.Different sectors have different contributions to make towards solving specific health problems. In each case, the profile, interests, incentives, and relationships of key individuals and sectors must be mapped and analysed to inform the design of approaches and systems to tackle a shared problem.Collaborative and distributed leadership is key for effective governance of multisectoral action, with a need to build leadership capacity across sectors and levels of government and cultivate champions in different sectors who can agree on common objectives.Important ways forward to support countries to take a multisectoral approach for health include ensuring that the universal health coverage agenda addresses the capacity of the health sector to work with other sectors, learning from multisectoral efforts that do not involve the health sector, improving the capacity of global institutions to support countries in undertaking multisectoral action, and developing a clear implementation research agenda for multisectoral action for health.

Campylobacter spp. commonly cause gastrointestinal illness in humans. Poultry meats have long been considered the predominant source of these infections, but few in-depth Campylobacter source attribution studies have been completed. We analyzed more than 1,300 Campylobacter isolates recovered from a number of animal and food sources, including dairy and beef cattle, pigs, poultry, and retail poultry meat, and compared them with Campylobacter isolates recovered from human clinical samples. Each isolate was subtyped using pulsed-field gel electrophoresis (PFGE) with SmaI and queried against the Centers for Disease Control and Prevention PulseNet database to identify human isolates with indistinguishable patterns. Half (49.5%) of the PFGE patterns from poultry animal and retail meat isolates were indistinguishable from patterns of at least one human isolate. Among the isolates from beef and dairy cows, 56.6 and 65.0%, respectively, of their PFGE patterns were indistinguishable from those of human isolates. Only a small portion of the PFGE patterns of Campylobacter isolated from pigs (9.5%) were found to have PFGE patterns in common with human isolates. These data imply that cattle may be larger contributors to Campylobacter infections than previously recognized and help further our understanding of potential sources of human campylobacteriosis.

Monoclonal antibodies targeting PD- 1 (nivolumab and pembrolizumab) and PD-L1 (avelumab, atezolizumab, and durvalumab) are FDA approved for the treatment of metastatic cancers. Immune-related adverse events (irAEs) are autoimmune diseases which occur following treatment with PD-1 and PD-L1 inhibitors. Neurological irAEs following PD-L1 blockade are rarely reported, and mainly consist of Myasthenia Gravis (MG) and myositis. MG induced by avelumab has not been previously reported. We present a 74-year-old female with stage IV ovarian adenocarcinoma who entered a clinical trial involving avelumab and entinostat after disease progression following extensive resection, chemotherapy with doxorubicin, then docetaxol and cisplatin. She developed progressive generalized weakness and fatigue two months after entering the trial, leading to respiratory weakness and hypoventilation, and resulting in intubation and mechanical ventilation. Exam showed head-drop, diplopia, and proximal > distal upper and lower limb weakness. Antibodies to acetylcholine receptors, muscle specific tyrosine kinase, and voltage-gated calcium channels (P/Q and N type) were negative, and serum creatine kinase was normal. Repetitive nerve stimulation showed significant decrement in the left spinal accessory nerve motor response with recording at trapezius. She was diagnosed with seronegative MG and was treated with IVIG, 2 grams/kg divided over 4 days, prednisone 60 mg daily, and pyridostigmine, which resulted in successful extubation, followed by gradual resolution of MG symptoms over a period of 3 months. At approximately 2 year follow-up, she remains on maintenance IVIG at 0.5 grams/kg every 3 weeks, as she develops proximal lower limb and neck extension weakness at the end of each IVIG cycle. We conclude that MG is a potentially life threatening but treatable irAE associated with avelumab. Our case also adds to the accumulating literature of the role of PD-1 signaling in the pathogenesis of autoimmune diseases of the muscles and neuromuscular junctions.

The fragility of health systems has never been of greater interest—or importance—than at this moment, in the aftermath of the worst Ebola virus disease epidemic to date. The loss of life, massive social disruption, and collapse of even the most basic health-care services shows what happens when a crisis hits and health systems are not prepared. This did not happen only in west Africa— we saw it in Texas too: the struggle to provide a coherent response and manage public sentiment (which often manifests as fear) in a way that ensures that disease does not spread while also allowing day-to-day life to continue. In other words, we saw an absence of resilience. This Viewpoint puts forth a proposed framework for resilient health systems and the characteristics that defi ne them, informed by insights from other fi elds that have embraced resilience as a practice. Health system resilience can be defi ned as the capacity of health actors, institutions, and populations to prepare for and eff ectively respond to crises; maintain core functions when a crisis hits; and, informed by lessons learned during the crisis, reorganise if conditions require it. Health systems are resilient if they protect human life and produce good health outcomes for all during a crisis and in its aftermath. Resilient health systems can also deliver everyday benefi ts and positive health outcomes. This double benefi t—improved performance in both bad times and good—is what has been called “the resilience dividend”. Response to a crisis, be it a disease outbreak or other disruption resulting in a surge of demand for health care (eg, a natural disaster or a mass casualty event) needs both a vigorous public health response and a highly proactive and functioning health-care delivery system. These two systems must work in concert during a crisis—and indeed long before crisis strikes. Health-care systems are complex adaptive systems and resilience is an emergent property of the health system as a whole rather than a single dimension. Building resilience is thus context-dependent and iterative, needing advance assessments of system capacities and weaknesses, investments in vulnerable components of the system before a crisis, reinforcements during the emergency, and review of performance after a crisis. Resilience is not a static construct—for example, the rapid pace of recovery from crisis is a cardinal measure of success. The Ebola epidemic has illustrated that several preconditions for resilience were lacking. The fi rst of these preconditions is recognition of the global nature of severe health crises and clarity about the roles of actors at all levels of the global health system. Although national governments are fundamentally responsible for their health systems, they need the capacity to mobilise the full range of local actors and to rapidly draw on external resources if necessary. The need for a global resilience network is both a moral imperative and a recognition of the fact that pathogens do not respect borders. Shocks to the health system of one country can reverberate across regions and the world. Health system resilience is thus a global public good and needs a collective response from the global community. Funding for this response can come from traditional domestic and donor sources or, as recently suggested, a new international health systems fund to which all countries contribute. A second precondition is a legal and policy foundation to guide the response and establish accountability. The implementation of International Health Regulations, which call on countries to build core public health capacities and establish a means of coordinating a response to health emergencies with regional and global partners, is a prerequisite for eff ective emergency response. Additionally, legislation that clarifi es the authority of public health agencies and the roles and responsibilities of private and public health actors is needed as are policies for involving the private and voluntary sector in the response and allowing fl exibility in sharing and reallocating resources across the health system. Third, there is a need for a strong and committed health workforce, characterised by health personnel who show up for work that might be diffi cult and dangerous. Establishing such a workforce begins with training and deployment of a suffi cient number of doctors, nurses, managers, and outreach workers—a colossal task in a country such as Liberia with a population of 3·5 million people and fewer than 100 doctors—but also building and banking a stock of social capital in the health system before crisis strikes. Just as strong social capital in communities promotes individual psychological resilience after mass trauma, social capital in the health system promotes system-wide recovery from crisis. In the health system context, social capital has two dimensions: a sense of worth, community, and responsibility among health actors (clinicians, managers, engineers, outreach workers) and an inclusive and robust community engagement with the health system. Health systems that earn the trust and support of the population and local political leaders by reliably providing high-quality services before crisis have a powerful resilience advantage. Strong management of district level health systems is key to gaining that trust. Diverse fi elds such as ecology, engineering, complex adaptive systems, psychology, and public health have produced resilience frameworks. The Rockefeller Foundation has developed substantial data about resilient Lancet 2015; 385: 1910–12

Summary Background The prodrug tenofovir alafenamide is associated with improved renal and bone safety compared with tenofovir disoproxil fumarate. We aimed to assess safety, pharmacokinetics, and efficacy of this single-tablet, fixed-dose combination of elvitegravir, cobicistat, emtricitabine, and tenofovir alafenamide in HIV-infected, treatment-naive adolescents. Methods We did a 48 week, single-arm, open-label trial in treatment-naive adolescents with HIV from ten hospital clinics in South Africa, Thailand, Uganda, and the USA. Eligible participants were aged 12–18 years, with plasma HIV-1 RNA of at least 1000 copies per mL, a CD4 count of at least 100 cells per μL, and estimated glomerular filtration rate of at least 90 mL/min per 1·73 m 2 by the Schwartz formula, bodyweight of at least 35 kg, and an HIV-1 genotype with sensitivity to elvitegravir, emtricitabine, and tenofovir. Participants received a single-tablet regimen once per day with food (administered by their parent or carer) containing 150 mg elvitegravir, 150 mg cobicistat, 200 mg emtricitabine, and 10 mg tenofovir alafenamide. Study visits to the clinic occurred at weeks 1, 2, 4, 8, 12, 16, 24, 32, 40, and 48. The coprimary endpoints were the pharmacokinetic parameters of area under the curve (AUC) concentration at the end of the dosing interval (AUC tau ) for elvitegravir and the AUC from time zero to the last quantifiable concentration (AUC last ) for tenofovir alafenamide, incidence of treatment-emergent serious adverse events, and all adverse events that emerged after treatment started (including data for bone mineral density). All participants who received one dose of study drug were included in the primary and safety analyses. This trial is registered with ClinicalTrials.gov, number NCT01854775. Findings Between May 22, 2013, and July 22, 2014, we enrolled 50 participants, and all 50 received the assigned treatment; 24 participated in the intensive pharmacokinetic assessment. 48 patients completed the 48 weeks of treatment; two discontinued (one withdrew consent at week 8, one was lost to follow-up at week 12). The regimen was well tolerated and no discontinuations related to adverse events occurred. The mean AUC tau for elvitegravir was 23 840 ng × h per mL (coefficient of variation [CV] 25·5%), and the mean AUC last for tenofovir alafenamide was 189 ng × h per mL (CV 55·8%). Four participants (8%) had a serious adverse event, one of which (intermediate uveitis) was deemed related to the study regimen but resolved without treatment interruption. The most common study drug-related adverse events were nausea (in ten participants), abdominal pain (in six), and vomiting (in five). Exposures to the elvitegravir, cobicistat, emtricitabine, and tenofovir alafenamide regimen were similar to those previously noted in adults. Interpretation The elvitegravir, cobicistat, emtricitabine, and tenofovir alafenamide regimen was well tolerated and achieved component plasma pharmacokinetic exposures similar to those in adults. Although non-comparative with a small sample size, these data support the use of this regimen in HIV-infected adolescents and its timely assessment in younger children. Funding Gilead Sciences.

There is an increasing interest within both organisational and social contexts in virtual worlds and virtual reality platforms. Virtual worlds are highly graphical systems in which avatars interact with each other, and almost every event and conversation is logged and recorded. This presents new challenges for qualitative researchers in information systems. This paper addresses the challenges of analyzing the huge amounts of qualitative data that can be obtained from virtual worlds (both images and text). It addresses how images might be used in qualitative studies of virtual worlds, and proposes a new way to analyze textual data using a qualitative software tool called Leximancer. This paper illustrates these methods using a study of a social movement in a virtual world.

Health system resilience begins with measurement of critical capacities ahead of crisis say Margaret E Kruk and colleagues The 2014 west African Ebola epidemic shone a harsh light on the health systems of Guinea, Liberia, and Sierra Leone. While decades of domestic and international investment had contributed to substantial progress on the Millennium Development Goals,12 national health systems remained weak and were unable to cope with the epidemic. Routine care of the population also deteriorated during the outbreak.1234 Surveillance systems did not function effectively, allowing Ebola to spread within and between the countries. Global institutions were slow to respond to the crisis, squandering an opportunity to stem its course.567 Since then, diverse panels of experts have pointed to political and technical deficiencies in multilateral organisations in tackling health crises.891011 These reports have noted that the first line of defence against future pandemics is an effective national health system. They have also called for better measurement of public health capacity, and investment to build resilient health systems—systems that can withstand health shocks while maintaining routine functions.10 The issue of how global bodies can support countries in withstanding future health shocks is playing out now in the election of the new director general of WHO, with several candidates making health system resilience part of their election planks. Based on recent literature, this paper defines health system resilience as “the capacity of health actors, institutions, and populations to prepare for and effectively respond to crises; maintain core functions when a crisis hits; and, informed by lessons learnt during the crisis, reorganise if conditions require it.”12 Health system resilience is relevant in all countries facing health shocks—whether sudden (Ebola, earthquakes, terror attacks, refugees), slower moving (new pathogens such as …

Carrageenan-induced inflammation, though extensively used as a model of local inflammation, has not been fully characterized on the genomic and proteomic level. The goal of this study was to develop an in vitro carrageenan model of local inflammation for use in studies of novel non-steroidal anti-inflammatory drug efficacy. We evaluated lambda carrageenan (l-CGN) in vitro and its capacity to affect previously identified inflammatory biomarkers in swine, using swine whole blood cultures. Blood was collected from 9 male Yorkshire barrows, cultured in vitro. The blood was divided into two groups (stimulated and unstimulated) per each time point. Unstimulated blood was used as control to compare samples treated with l-CGN. Blood was then separated into two fractions, serum collection for ELISA analysis and white blood cells for RNA extraction per time point. We found that l-CGN increased the production MCP-1 in vitro, both on the proteomics level and the genomic level. Conversely, TNF-a and IL-6 demonstrated a decreased or no change in cytokine production when measured via ELISA. Phosphokinase-3, Thrombospondin-1 and Alveolar Macrophage Chemotactic -II showed increased gene expression when normalized to GAPDH. These results also demonstrate that carrageenan initiates an inflammatory response that differs from a typical endotoxin effect, such as LPS stimulation in terms of the pathways and gene products altered.

Non-steroidal anti-inflammatory drugs (NSAIDs) control inflammation through reductions in Prostaglandin E2 (PGE2) levels. Recent work by this laboratory identified NSAID-induced changes in inflammatory gene expression that appear to be independent of changes in PGE2; however, the molecular mechanism is unknown. Our goal is to determine whether NSAIDs affect gene expression via inhibition of NFkβ. Knowledge of NSAID gene control will provide a mechanism for screening novel drugs claiming anti-inflammatory properties. RAW 264.7 cells transfected with a plasmid under NFkβ control to express Secreted Alkaline Phosphatase (SEAP) were pre-treated with NSAID concentrations ranging from 10-3 to 10-7 M and stimulated with 1μg/mL LPS for 24 hours. NSAIDs from the 5 major classes inhibited SEAP release at 10-3 M, while acetylsalicylic acid (ASA) and acetaminophen did not. HEK293 cells transfected with the same plasmid were pre-treated with flunixin, diclofenac, and ASA at concentrations from 10-3 to 10-7 M and stimulated with 10ng/mL TNFα for 24 hours. Diclofenac and flunixin decreased SEAP release at 10-3 M, whereas ASA failed to inhibit SEAP release. These results demonstrate that NSAIDs prevent SEAP release via NFkβ inhibition, suggesting a new target for novel NSAIDs. Experiments to confirm NSAID inhibition of NFkβ activation by measuring the release of p65 are underway. The results of this research may also provide a screening tool for novel NSAIDs.