Verifying Transcription Factor Regulation of ADV-1 Protein Rhythms through Complementation

Roughly half of the eukaryotic genome is under the control of a circadian clock at the level of transcript abundance, with rhythmic transcripts peaking at all possible phases of the day. However, the mechanism for phase regulation is not well understood. To determine how phase is regulated in Neurospora crassa, ChIP-seq was performed on the morning active transcription factor (TF) and core oscillator component, the White Collar Complex (WCC). The direct targets of WCC were found to be enriched for TFs. Further investigation of these first-tiered TFs revealed that they form a large and complex network made up of multiple feedback and feedforward loops. We hypothesized that this intricate TF network plays a role in phase regulation. To test this hypothesis, we focused our experiments on the network surrounding the first-tier TF, ADV-1. In preliminary data, we found that TF knockouts of CSP-1, CSP-2, SUB-1, and ZNF-21, altered the phase and/or period of ADV-1 protein rhythms. To confirm that these 4 TF knockouts, and not second site mutations, cause the changes in ADV-1 rhythms, genetic complementation will be performed. If the TF knockout causes the defect, then supplying the wild type gene back should rescue the ADV-1 rhythm back to wild type. This information will help us to validate which TFs in the network are necessary for generating proper phase control of ADV-1 protein rhythms

Verifying Transcription Factor Regulation of ADV-1 Protein Rhythms through Complementation

Roughly half of the eukaryotic genome is under the control of a circadian clock at the level of transcript abundance, with rhythmic transcripts peaking at all possible phases of the day. However, the mechanism for phase regulation is not well understood. To determine how phase is regulated in Neurospora crassa, ChIP-seq was performed on the morning active transcription factor (TF) and core oscillator component, the White Collar Complex (WCC). The direct targets of WCC were found to be enriched for TFs. Further investigation of these first-tiered TFs revealed that they form a large and complex network made up of multiple feedback and feedforward loops. We hypothesized that this intricate TF network plays a role in phase regulation. To test this hypothesis, we focused our experiments on the network surrounding the first-tier TF, ADV-1. In preliminary data, we found that TF knockouts of CSP-1, CSP-2, SUB-1, and ZNF-21, altered the phase and/or period of ADV-1 protein rhythms. To confirm that these 4 TF knockouts, and not second site mutations, cause the changes in ADV-1 rhythms, genetic complementation will be performed. If the TF knockout causes the defect, then supplying the wild type gene back should rescue the ADV-1 rhythm back to wild type. This information will help us to validate which TFs in the network are necessary for generating proper phase control of ADV-1 protein rhythms

MagneToRE: Mapping the 3-D Magnetic Structure of the Solar Wind Using a Large Constellation of Nanosatellites

Unlike the vast majority of astrophysical plasmas, the solar wind is accessible to spacecraft, which for decades have carried in-situ instruments for directly measuring its particles and fields. Though such measurements provide precise and detailed information, a single spacecraft on its own cannot disentangle spatial and temporal fluctuations. Even a modest constellation of in-situ spacecraft, though capable of characterizing fluctuations at one or more scales, cannot fully determine the plasma’s 3-D structure. We describe here a concept for a new mission, the Magnetic Topology Reconstruction Explorer (MagneToRE), that would comprise a large constellation of in-situ spacecraft and would, for the first time, enable 3-D maps to be reconstructed of the solar wind’s dynamic magnetic structure. Each of these nanosatellites would be based on the CubeSat form-factor and carry a compact fluxgate magnetometer. A larger spacecraft would deploy these smaller ones and also serve as their telemetry link to the ground and as a host for ancillary scientific instruments. Such an ambitious mission would be feasible under typical funding constraints thanks to advances in the miniaturization of spacecraft and instruments and breakthroughs in data science and machine learning

BHPR research: qualitative1. Complex reasoning determines patients' perception of outcome following foot surgery in rheumatoid arhtritis

Background: Foot surgery is common in patients with RA but research into surgical outcomes is limited and conceptually flawed as current outcome measures lack face validity: to date no one has asked patients what is important to them. This study aimed to determine which factors are important to patients when evaluating the success of foot surgery in RA Methods: Semi structured interviews of RA patients who had undergone foot surgery were conducted and transcribed verbatim. Thematic analysis of interviews was conducted to explore issues that were important to patients. Results: 11 RA patients (9 ♂, mean age 59, dis dur = 22yrs, mean of 3 yrs post op) with mixed experiences of foot surgery were interviewed. Patients interpreted outcome in respect to a multitude of factors, frequently positive change in one aspect contrasted with negative opinions about another. Overall, four major themes emerged. Function: Functional ability & participation in valued activities were very important to patients. Walking ability was a key concern but patients interpreted levels of activity in light of other aspects of their disease, reflecting on change in functional ability more than overall level. Positive feelings of improved mobility were often moderated by negative self perception ("I mean, I still walk like a waddling duck”). Appearance: Appearance was important to almost all patients but perhaps the most complex theme of all. Physical appearance, foot shape, and footwear were closely interlinked, yet patients saw these as distinct separate concepts. Patients need to legitimize these feelings was clear and they frequently entered into a defensive repertoire ("it's not cosmetic surgery; it's something that's more important than that, you know?”). Clinician opinion: Surgeons' post operative evaluation of the procedure was very influential. The impact of this appraisal continued to affect patients' lasting impression irrespective of how the outcome compared to their initial goals ("when he'd done it ... he said that hasn't worked as good as he'd wanted to ... but the pain has gone”). Pain: Whilst pain was important to almost all patients, it appeared to be less important than the other themes. Pain was predominately raised when it influenced other themes, such as function; many still felt the need to legitimize their foot pain in order for health professionals to take it seriously ("in the end I went to my GP because it had happened a few times and I went to an orthopaedic surgeon who was quite dismissive of it, it was like what are you complaining about”). Conclusions: Patients interpret the outcome of foot surgery using a multitude of interrelated factors, particularly functional ability, appearance and surgeons' appraisal of the procedure. While pain was often noted, this appeared less important than other factors in the overall outcome of the surgery. Future research into foot surgery should incorporate the complexity of how patients determine their outcome Disclosure statement: All authors have declared no conflicts of interes

Case Reports1. A Late Presentation of Loeys-Dietz Syndrome: Beware of TGFβ Receptor Mutations in Benign Joint Hypermobility

Background: Thoracic aortic aneurysms (TAA) and dissections are not uncommon causes of sudden death in young adults. Loeys-Dietz syndrome (LDS) is a rare, recently described, autosomal dominant, connective tissue disease characterized by aggressive arterial aneurysms, resulting from mutations in the transforming growth factor beta (TGFβ) receptor genes TGFBR1 and TGFBR2. Mean age at death is 26.1 years, most often due to aortic dissection. We report an unusually late presentation of LDS, diagnosed following elective surgery in a female with a long history of joint hypermobility. Methods: A 51-year-old Caucasian lady complained of chest pain and headache following a dural leak from spinal anaesthesia for an elective ankle arthroscopy. CT scan and echocardiography demonstrated a dilated aortic root and significant aortic regurgitation. MRA demonstrated aortic tortuosity, an infrarenal aortic aneurysm and aneurysms in the left renal and right internal mammary arteries. She underwent aortic root repair and aortic valve replacement. She had a background of long-standing joint pains secondary to hypermobility, easy bruising, unusual fracture susceptibility and mild bronchiectasis. She had one healthy child age 32, after which she suffered a uterine prolapse. Examination revealed mild Marfanoid features. Uvula, skin and ophthalmological examination was normal. Results: Fibrillin-1 testing for Marfan syndrome (MFS) was negative. Detection of a c.1270G > C (p.Gly424Arg) TGFBR2 mutation confirmed the diagnosis of LDS. Losartan was started for vascular protection. Conclusions: LDS is a severe inherited vasculopathy that usually presents in childhood. It is characterized by aortic root dilatation and ascending aneurysms. There is a higher risk of aortic dissection compared with MFS. Clinical features overlap with MFS and Ehlers Danlos syndrome Type IV, but differentiating dysmorphogenic features include ocular hypertelorism, bifid uvula and cleft palate. Echocardiography and MRA or CT scanning from head to pelvis is recommended to establish the extent of vascular involvement. Management involves early surgical intervention, including early valve-sparing aortic root replacement, genetic counselling and close monitoring in pregnancy. Despite being caused by loss of function mutations in either TGFβ receptor, paradoxical activation of TGFβ signalling is seen, suggesting that TGFβ antagonism may confer disease modifying effects similar to those observed in MFS. TGFβ antagonism can be achieved with angiotensin antagonists, such as Losartan, which is able to delay aortic aneurysm development in preclinical models and in patients with MFS. Our case emphasizes the importance of timely recognition of vasculopathy syndromes in patients with hypermobility and the need for early surgical intervention. It also highlights their heterogeneity and the potential for late presentation. Disclosures: The authors have declared no conflicts of interes

Recommendations for Addressing Priority Io Science in the Next Decade

Io is a priority destination for solar system exploration. The scope and importance of science questions at Io necessitates a broad portfolio of research and analysis, telescopic observations, and planetary missions - including a dedicated New Frontiers class Io mission

The Science Case for Io Exploration

Io is a priority destination for solar system exploration, as it is the best natural laboratory to study the intertwined processes of tidal heating, extreme volcanism, and atmosphere-magnetosphere interactions. Io exploration is relevant to understanding terrestrial worlds (including the early Earth), ocean worlds, and exoplanets across the cosmos

Effect of angiotensin-converting enzyme inhibitor and angiotensin receptor blocker initiation on organ support-free days in patients hospitalized with COVID-19

IMPORTANCE Overactivation of the renin-angiotensin system (RAS) may contribute to poor clinical outcomes in patients with COVID-19. Objective To determine whether angiotensin-converting enzyme (ACE) inhibitor or angiotensin receptor blocker (ARB) initiation improves outcomes in patients hospitalized for COVID-19. DESIGN, SETTING, AND PARTICIPANTS In an ongoing, adaptive platform randomized clinical trial, 721 critically ill and 58 non–critically ill hospitalized adults were randomized to receive an RAS inhibitor or control between March 16, 2021, and February 25, 2022, at 69 sites in 7 countries (final follow-up on June 1, 2022). INTERVENTIONS Patients were randomized to receive open-label initiation of an ACE inhibitor (n = 257), ARB (n = 248), ARB in combination with DMX-200 (a chemokine receptor-2 inhibitor; n = 10), or no RAS inhibitor (control; n = 264) for up to 10 days. MAIN OUTCOMES AND MEASURES The primary outcome was organ support–free days, a composite of hospital survival and days alive without cardiovascular or respiratory organ support through 21 days. The primary analysis was a bayesian cumulative logistic model. Odds ratios (ORs) greater than 1 represent improved outcomes. RESULTS On February 25, 2022, enrollment was discontinued due to safety concerns. Among 679 critically ill patients with available primary outcome data, the median age was 56 years and 239 participants (35.2%) were women. Median (IQR) organ support–free days among critically ill patients was 10 (–1 to 16) in the ACE inhibitor group (n = 231), 8 (–1 to 17) in the ARB group (n = 217), and 12 (0 to 17) in the control group (n = 231) (median adjusted odds ratios of 0.77 [95% bayesian credible interval, 0.58-1.06] for improvement for ACE inhibitor and 0.76 [95% credible interval, 0.56-1.05] for ARB compared with control). The posterior probabilities that ACE inhibitors and ARBs worsened organ support–free days compared with control were 94.9% and 95.4%, respectively. Hospital survival occurred in 166 of 231 critically ill participants (71.9%) in the ACE inhibitor group, 152 of 217 (70.0%) in the ARB group, and 182 of 231 (78.8%) in the control group (posterior probabilities that ACE inhibitor and ARB worsened hospital survival compared with control were 95.3% and 98.1%, respectively). CONCLUSIONS AND RELEVANCE In this trial, among critically ill adults with COVID-19, initiation of an ACE inhibitor or ARB did not improve, and likely worsened, clinical outcomes. TRIAL REGISTRATION ClinicalTrials.gov Identifier: NCT0273570