Search

Plasmacytoid dendritic cells (pDCs) are first responders to tissue damage and interact with wound CD4 T-cells to orchestrate tissue repair. In diabetes, the role of pDCs and their subsequent influence on wound CD4 T-cells and inflammation during tissue repair is unknown. Using human diabetic wound single cell sequencing, we identified that human wound CD4 T cells are primarily the Th17 phenotype. Thus, we hypothesized that pDCs in the wounds may regulate this inflammatory T cell phenotype, contributing to chronic inflammation. Although pDCs exist in low numbers in the wounds, we isolated wound pDCs from diabetic and control mice and examined them for inflammatory cytokine expression. We found that diabetic pDC produced significantly more IL6 (amongst other inflammatory cytokines) compared to controls. This is important as IL6 has been shown to skew CD4 T-cells towards a Th17 phenotype in tissues. Upon co-culture of diabetic pDCs with naïve CD4 T-cells, a significant increase in Th17 cells was observed by intracellular flow cytometry analysis. In order to examine the increased IL6 from diabetic pDCs we isolated diabetic and control wound pDCs and performed an epigenetic superarray. We identified that histone demethylase Jarid1C was significantly decreased in diabetic pDCs compared to controls. We then isolated diabetic pDCs and performed a ChIP analysis on the IL6 NFkB binding sites in the promoter and identified an increase in tri-methylated lysine 4 on histone 3 (H3K4me3), a marker regulated by Jarid 1C and consistent with increased transcription of IL6. Taken together, this data suggests that in diabetes, pDCs are epigenetically altered to produce increased IL6 and contribute to increased Th17 and inflammation in diabetic wounds.

Abdominal aortic aneurysms (AAAs) are focal dilations of the aorta that develop from degenerative changes in the media and adventitia of the vessel. Ruptured AAAs have a mortality of up to 85%, thus it is important to identify patients with AAA at increased risk for rupture who would benefit from increased surveillance and/or surgical repair. Although the exact genetic and epigenetic mechanisms regulating AAA formation are not completely understood, Mendelian cases of AAA, which result from pathologic variants in a single gene, have helped provide a basic understanding of AAA pathophysiology. More recently, genome wide associated studies (GWAS) have identified additional variants, termed single nucleotide polymorphisms, in humans that may be associated with AAAs. While some variants may be associated with AAAs and play causal roles in aneurysm pathogenesis, it should be emphasized that the majority of SNPs do not actually cause disease. In addition to GWAS, other studies have uncovered epigenetic causes of disease that regulate expression of genes known to be important in AAA pathogenesis. This review describes many of these genetic and epigenetic contributors of AAAs, which altogether provide a deeper insight into AAA pathogenesis.

Significance The COVID-19 pandemic has disproportionately affected patients with comorbidities, namely, obesity and type 2 diabetes. Macrophages (Mφs) are a key innate immune cell primarily responsible for the harmful, hyperinflammatory “cytokine storm” in patients that develop severe COVID-19. We describe a mechanism for this Mφ-mediated cytokine storm in response to coronavirus. In response to coronavirus infection, expression of the chromatin-modifying enzyme, SETDB2, decreases in Mφs, leading to increased transcription of inflammatory cytokines. Further, we find SETDB2 is regulated by an interferon beta (IFNβ)/JaK/STAT3 mechanism, and that exogenous administration of IFNβ can reverse inflammation, particularly in diabetic Mφs via an increase in SETDB2. Together, these results suggest therapeutic targeting of the IFNβ/SETDB2 axis in diabetic patients with COVID-19 may decrease pathologic inflammation., COVID-19 induces a robust, extended inflammatory “cytokine storm” that contributes to an increased morbidity and mortality, particularly in patients with type 2 diabetes (T2D). Macrophages are a key innate immune cell population responsible for the cytokine storm that has been shown, in T2D, to promote excess inflammation in response to infection. Using peripheral monocytes and sera from human patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and a murine hepatitis coronavirus (MHV-A59) (an established murine model of SARS), we identified that coronavirus induces an increased Mφ-mediated inflammatory response due to a coronavirus-induced decrease in the histone methyltransferase, SETDB2. This decrease in SETDB2 upon coronavirus infection results in a decrease of the repressive trimethylation of histone 3 lysine 9 (H3K9me3) at NFkB binding sites on inflammatory gene promoters, effectively increasing inflammation. Mφs isolated from mice with a myeloid-specific deletion of SETDB2 displayed increased pathologic inflammation following coronavirus infection. Further, IFNβ directly regulates SETDB2 in Mφs via JaK1/STAT3 signaling, as blockade of this pathway altered SETDB2 and the inflammatory response to coronavirus infection. Importantly, we also found that loss of SETDB2 mediates an increased inflammatory response in diabetic Mϕs in response to coronavirus infection. Treatment of coronavirus-infected diabetic Mφs with IFNβ reversed the inflammatory cytokine production via up-regulation of SETDB2/H3K9me3 on inflammatory gene promoters. Together, these results describe a potential mechanism for the increased Mφ-mediated cytokine storm in patients with T2D in response to COVID-19 and suggest that therapeutic targeting of the IFNβ/SETDB2 axis in T2D patients may decrease pathologic inflammation associated with COVID-19.

Introduction: CD4+ T-cell activation is vital for normal wound repair but the factors that control T-cell activation in wounds in vivo are not clear. Our group and others have found increased Th17 activation in diabetic wounds resulting in increased IL-17a and pathologic inflammation that prevents tissue repair. Plasmacytoid dendritic cells (pDC) are antigen presenting cells that are present in early diabetic wound tissue and may play a key role in modulating CD4+ T-cell phenotype. Hence, we hypothesized that diabetic pDCs may influence wound CD4+ T-cells towards Th17 T-cell expansion. Methods: Wild type C57BL/6 mice were fed normal chow diet (13.5% kcal fat; LabDiet) or high fat diet chow (60% kcal fat; Research Diets) for 12–14 weeks to generate the diet-induced obesity (DIO) model of glucose intolerance/insulin resistance. These mice were subsequently wounded, and wound plasmacytoid dendritic cells harvested on day 1 and day 3. These cells were co-cultured with naïve CD4+ T-cells for 3 days, after which T-cell phenotype was determined by flow cytometry. Additionally, pDC in wounds 1-day post wounding were examined by quantitative PCR for cytokine production. Results: Following exposure to DIO pDCs, wild type activated CD4+ T-cells were activated towards a Th17 phenotype via significant increases in TGFb CD4+ T-cell activation and may act to increase inflammation in diabetic wounds.

COVID-19 induces a robust inflammatory ‘cytokine storm’ that contributes to an increased morbidity and mortality, particularly in patients with type 2 diabetes (T2D). Macrophages are a key innate immune cell population responsible for the ‘cytokine storm’ that has been shown in T2D to promote excess inflammation in response to infection. Using sera from human patients with SARS-CoV-2 and a murine hepatitis coronavirus (MHV-A59) (an established murine model of SARS), we identified that coronavirus induces an increased Mϕ-mediated inflammatory response due to a coronavirus-induced decrease in the histone methyltransferase, SETDB2. This decrease in SETDB2 upon coronavirus infection results in a decrease of the repressive trimethylation of histone 3 lysine 9 (H3K9me3) at NFkB binding sites on inflammatory gene promoters, increasing inflammation. Mϕs with a myeloid-specific deletion of SETDB2 displayed increased inflammation following coronavirus infection. Further, IFNβ directly regulates SETDB2 in Mϕs via JaK1/STAT3 signaling, as blockade of this pathway altered SETDB2 and the inflammatory response to coronavirus infection. Importantly, we also found that SETDB2 mediates a heightened inflammatory response in diabetic Mϕs in response to coronavirus infection. Treatment of coronavirus-infected diabetic Mϕs with IFNβ reversed the inflammatory cytokine production via upregulation of SETDB2/H3K9me3 on inflammatory gene promoters. Together, these results describe a potential mechanism for the increased Mϕ-mediated ‘cytokine storm’ in patients with T2D in response to COVID-19 and suggest that therapeutic targeting of the IFNβ/SETDB2 axis in T2D patients may decrease pathologic inflammation associated with COVID-19.

Objective Colloid preloading diminishes post-spinal hypotension. However, whether colloid preloading is superior to crystalloid co-loading is uncertain. In this retrospective study, we compared the effects of a colloid preload versus a crystalloid co-load on vasopressor requirements and maternal haemodynamics among women undergoing elective caesarean delivery (CD) with spinal anaesthesia. Methods We extracted data from the medical records of 160 healthy women who underwent elective CD with spinal anaesthesia at an academic obstetric centre before and after an institutional fluid-loading protocol change. Patients received a 500 mL 6% hydroxyethyl starch preload or a 1000 mL crystalloid co-load. The primary outcome was the total phenylephrine dose administered from spinal block placement to delivery. Results Our cohort comprised 79 women in the colloid group and 77 women in the crystalloid group. The mean phenylephrine use was significantly lower in the colloid group than in the crystalloid group (489±403 μg vs. 647±464 μg, respectively, p=0.02). The maximal drop in systolic blood pressure was greater in the colloid group than in the crystalloid group (36±20 mmHg vs. 29±16 mmHg, respectively, p=0.02). There were no clinically significant differences between the groups in heart rate, blood loss, temperature and Apgar scores. Conclusion Vasopressor use was lower in colloid preloading than in crystalloid co-loading. However, differences in all outcome measures were minimal and likely clinically insignificant, suggesting that both fluid-loading techniques are appropriate to use for the prevention of spinal hypotension in women undergoing CD.

The National Aeronautics and Space Administration (NASA) invests millions of dollars in spacecraft and ground system development, and in mission operations in the pursuit of scientific knowledge of the universe. In recent years, NASA sent a probe to Mars to study the Red Planet's upper atmosphere, obtained high resolution images of Pluto, and it is currently preparing to find new exoplanets, rendezvous with an asteroid, and bring a sample of the asteroid back to Earth for analysis. The success of these missions is enabled by mission assurance. In turn, mission assurance is backed by information assurance. The information systems supporting NASA missions must be reliable as well as secure. NASA - like every other U.S. Federal Government agency - is required to manage the security of its information systems according to federal mandates, the most prominent being the Federal Information Security Management Act (FISMA) of 2002 and the legislative updates that followed it. Like the management of enterprise information technology (IT), federal information security management takes a "one-size fits all" approach for protecting IT systems. While this approach works for most organizations, it does not effectively translate into security of highly specialized systems such as those supporting NASA missions. These systems include command and control (C&C) systems, spacecraft and instrument simulators, and other elements comprising the ground segment. They must be carefully configured, monitored and maintained, sometimes for several years past the missions' initially planned life expectancy, to ensure the ground system is protected and remains operational without any compromise of its confidentiality, integrity and availability. Enterprise policies, processes, procedures and products, if not effectively tailored to meet mission requirements, may not offer the needed security for protecting the information system, and they may even become disruptive to mission operations. Certain protective measures for the general enterprise may not be as efficient within the ground segment. This is what the authors have concluded through observations and analysis of patterns identified from the various security assessments performed on NASA missions such as MAVEN, OSIRIS-REx, New Horizons and TESS, to name a few. The security audits confirmed that the framework for managing information system security developed by the National Institute of Standards and Technology (NIST) for the federal government, and adopted by NASA, is indeed effective. However, the selection of the technical, operational and management security controls offered by the NIST model - and how they are implemented - does not always fit the nature and the environment where the ground system operates in even though there is no apparent impact on mission success. The authors observed that unfit controls, that is, controls that are not necessarily applicable or sufficiently effective in protecting the mission systems, are often selected to facilitate compliance with security requirements and organizational expectations even if the selected controls offer minimum or non-existent protection. This paper identifies some of the standard security controls that can in fact protect the ground system, and which of them offer little or no benefit at all. It offers multiple scenarios from real security audits in which the controls are not effective without, of course, disclosing any sensitive information about the missions assessed. In addition to selection and implementation of controls, the paper also discusses potential impact of recent legislation such as the Federal Information Security Modernization Act (FISMA) of 2014 - aimed at the enterprise - on the ground system, and offers other recommendations to Information System Owners (ISOs).

Objective Spinal manipulative therapy (SMT) has been reported to successfully treat hypertension (HTN). The purpose of this study was to perform a qualitative literature review on the efficacy of SMT for treating HTN. Methods The literature was systematically searched in PubMed, Medline, Allied and Complementary Medicine Database, Cumulative Index to Nursing and Allied Health Literature, and Index of Chiropractic Literature. Included articles were rated for bias using the Cochrane Collaboration's tool for assessing risk of bias. Studies reporting differing methodologies, types of SMT, frequency of treatment, and time of follow-up were considered too dissimilar for meta-analysis. Results Of 208 articles identified, 10 were selected as relevant and were assessed. Risk of bias scores revealed 2 studies with low risk, 3 studies with unclear risk, and 5 studies with high risk. The maximum improvement observed in any SMT group, in low risk of bias studies was −9.7 (95% confidence interval [CI], −21.1 to 1.8) systolic improvement and −9.0 (95% CI, −16.8 to −1.2) diastolic; and in unclear risk of bias studies, it was −17.2 (95% CI, −20.7 to −13.7) systolic and −13.0 (95% CI, −15.4 to −10.6) diastolic. Statistically significant decreases in blood pressure were not observed in clinical trials with low bias when SMT was compared with effleurage massage and a 5-minute wait. The studies with more risk of bias showed a greater treatment effect. Conclusion There is currently a lack of low bias evidence to support the use of SMT as a therapy for the treatment of HTN. Future investigations may clarify if SMT is effective for treating HTN, either by itself or as an adjunctive therapy, and by which physiologic mechanism this occurs.

Activation of RhoA in vascular smooth muscle cells (SMC) has been linked to vasoconstrictor-induced hypertension (HTN), but the relevance of this pathway to human disease was undetermined. We identified GRAF3 as a RhoA-GAP expressed specifically in SMC in mice and humans and reported that global GRAF3-deficient mice exhibited significant basal HTN (+ 25 mm Hg) that was fully reversed by treatment with a Rho-kinase inhibitor (Nature Comm. 2013;4:2910). Importantly, we recently created a tamoxifen inducible SMC-GRAF3 re-expression model which resulted in a near complete reversal of MAP (from 123 mmHg to 95 mm Hg), indicating that GRAF3's ability to limit RhoA activity in SMC is required for the maintenance of normal BP. Given that a BP-associated locus within the GRAF3 gene was identified by Genome Wide Association, we next sought to characterize the mechanisms that control GRAF3 expression. Through the use of a series of siRNA-dependent approaches in cultured SMC, we found that SMC-specific expression of GRAF3 is mediated by the RhoA/MRTF-A/SRF-signaling axis. Interestingly, RhoA is known to be activated by mechanical forces and we found that GRAF3 expression was significantly increased (8-fold) in vessels subjected to pathological stress. The finding that GRAF3 expression was significantly increased in two separate hypertensive animal models relative to their littermate controls provides further evidence that GRAF3 expression is regulated as part of an auto-regulatory negative feedback loop to inhibit RhoA activity and SMC tone. Interestingly, the BP associated locus maps to the 80Kb first intron of the GRAF3 gene and we found that the minor GRAF3 allele that decreases BP was associated with a significant increase in GRAF3 mRNA in human tibial artery samples (3-fold increase; N=123; p=1e-10 ). We have identified regulatory elements within the hypertensive locus that exhibit SMC-selective transcriptional activity and have shown that a minor allele variation within one of these elements significantly increased transcriptional activity. Our studies add significantly to our understanding of the development and pathophysiologic consequences of hypertension and may lead to novel and perhaps individualized approaches to its treatment.

We have recently shown that the Rho-specific GAP, GRAF3, is specifically expressed in smooth muscle cells (SMC) in mouse and humans and negatively regulates blood pressure by limiting Rho-dependent SMC contractility. Genome wide association studies identified a hypertension-associated locus within the first intron of the GRAF3 gene (currently defined by SNPs rs633185 and rs604723) and demonstrated that the minor allele was significantly correlated with decreased blood pressure in patients. The aims of the current study were to identify transcription mechanisms that regulate SMC-specific GRAF3 expression and to test the hypothesis that SNPs within the GRAF3 hypertension locus affect blood pressure by altering GRAF3 expression and RhoA activity. Using DNaseI Hypersensitivity of chromatin structure, ChIP-seq approaches for previously implicated transcription factors, and information from sequence/regulatory databases, we identified two regions within the human GRAF3 gene that exhibited strong SMC-selective transcription activity in cultured cells. Importantly, the enhancer at +52 Kb contains SNP rs604723 and mutation of the major C allele to the minor T allele significantly increased enhancer activity from 20- to 40-fold over promoterless. The DNA surrounding the minor T allele conforms to the consensus SRF binding element at 8 out of 10 residues. Gel shift and targeted ChIP assays revealed preferential binding of SRF to the minor allele sequence, and the SRF co-factor, myocardin, significantly transactivated the minor but not the major allele enhancer (p

In NASA space flight missions, the Mission Operations Center (MOC) is often considered “the center of the (ground segment) universe,” at least by those involved with ground system operations. It is at and through the MOC that spacecraft is commanded and controlled, and science data acquired. This critical element of the ground system must be protected to ensure the confidentiality, integrity and availability of the information and information systems supporting mission operations. This paper identifies and highlights key information security aspects affecting MOCs that should be taken into consideration when reviewing and/or implementing protecting measures in and around MOCs. It stresses the need for compliance with information security regulation and mandates, and the need for the reduction of IT security risks that can potentially have a negative impact to the mission if not addressed. This compilation of key security aspects was derived from numerous observations, findings, and issues discovered by IT security audits the authors have conducted on NASA mission operations centers in the past few years. It is not a recipe on how to secure MOCs, but rather an insight into key areas that must be secured to strengthen the MOC, and enable mission assurance. Most concepts and recommendations in the paper can be applied to non-NASA organizations as well. Finally, the paper emphasizes the importance of integrating information security into the MOC development life cycle as configuration, risk and other management processes are tailored to support the delicate environment in which mission operations take place.

As the first interplanetary mission managed by the NASA Goddard Space Flight Center, the Mars Atmosphere and Volatile EvolutioN (MAVEN) had three IT security goals for its ground system: COMPLIANCE, (IT) RISK REDUCTION, and COST REDUCTION. In a multi-organizational environment in which government, industry and academia work together in support of the ground system and mission operations, information security governance, risk management, and compliance (GRC) becomes a challenge as each component of the ground system has and follows its own set of IT security requirements. These requirements are not necessarily the same or even similar to each other's, making the auditing of the ground system security a challenging feat. A combination of standards-based information security management based on the National Institute of Standards and Technology (NIST) Risk Management Framework (RMF), due diligence by the Mission's leadership, and effective collaboration among all elements of the ground system enabled MAVEN to successfully meet NASA's requirements for IT security, and therefore meet Federal Information Security Management Act (FISMA) mandate on the Agency. Throughout the implementation of GRC on MAVEN during the early stages of the mission development, the Project faced many challenges some of which have been identified in this paper. The purpose of this paper is to document these challenges, and provide a brief analysis of the lessons MAVEN learned. The historical information documented herein, derived from an internal pre-launch lessons learned analysis, can be used by current and future missions and organizations implementing and auditing GRC.