Multiphase coalescence mediates Hippo pathway activation.

The function of biomolecular condensates is often restricted by condensate dissolution. Whether condensates can be suppressed without condensate dissolution is unclear. Here, we show that upstream regulators of the Hippo signaling pathway form functionally antagonizing condensates, and their coalescence into a common phase provides a mode of counteracting the function of biomolecular condensates without condensate dissolution. Specifically, the negative regulator SLMAP forms Hippo-inactivating condensates to facilitate pathway inhibition by the STRIPAK complex. In response to cell-cell contact or osmotic stress, the positive regulators AMOT and KIBRA form Hippo-activating condensates to facilitate pathway activation. The functionally antagonizing SLMAP and AMOT/KIBRA condensates further coalesce into a common phase to inhibit STRIPAK function. These findings provide a paradigm for restricting the activity of biomolecular condensates without condensate dissolution, shed light on the molecular principles of multiphase organization, and offer a conceptual framework for understanding upstream regulation of the Hippo signaling pathway.

Control of stem cell renewal and fate by YAP and TAZ.

Complex physiological processes control whether stem cells self-renew, differentiate or remain quiescent. Two decades of research have placed the Hippo pathway, a highly conserved kinase signalling cascade, and its downstream molecular effectors YAP and TAZ at the nexus of this decision. YAP and TAZ translate complex biological cues acting on stem cells - from mechanical forces to cellular metabolism - into genome-wide effects to mediate stem cell functions. While aberrant YAP/TAZ activity drives stem cell dysfunction in ageing, tumorigenesis and disease, therapeutic targeting of Hippo signalling and YAP/TAZ can boost stem cell activity to enhance regeneration. In this Review, we discuss how YAP/TAZ control the self-renewal, fate and plasticity of stem cells in different contexts, how dysregulation of YAP/TAZ in stem cells leads to disease, and how therapeutic modalities targeting YAP/TAZ may benefit regenerative medicine and cancer therapy.

WWTR1(TAZ)-CAMTA1 reprograms endothelial cells to drive epithelioid hemangioendothelioma.

Epithelioid hemangioendothelioma (EHE) is a poorly understood and devastating vascular cancer. Sequencing of EHE has revealed a unique gene fusion between the Hippo pathway nuclear effector TAZ (WWTR1) and the brain-enriched transcription factor CAMTA1 in ∼90% of cases. However, it remains unclear whether the TAZ-CAMTA1 gene fusion is a driver of EHE, and potential targeted therapies are unknown. Here, we show that TAZ-CAMTA1 expression in endothelial cells is sufficient to drive the formation of vascular tumors with the distinctive features of EHE, and inhibition of TAZ-CAMTA1 results in the regression of these vascular tumors. We further show that activated TAZ resembles TAZ-CAMTA1 in driving the formation of EHE-like vascular tumors, suggesting that constitutive activation of TAZ underlies the pathological features of EHE. We show that TAZ-CAMTA1 initiates an angiogenic and regenerative-like transcriptional program in endothelial cells, and disruption of the TAZ-CAMTA1-TEAD interaction or ectopic expression of a dominant negative TEAD in vivo inhibits TAZ-CAMTA1-mediated transformation. Our study provides the first genetic model of a TAZ fusion oncoprotein driving its associated human cancer, pinpointing TAZ-CAMTA1 as the key driver and a valid therapeutic target of EHE.

Accelerated Aging after Traumatic Brain Injury: An ENIGMA Multi-Cohort Mega-Analysis.

OBJECTIVE: The long-term consequences of traumatic brain injury (TBI) on brain structure remain uncertain. Given evidence that a single significant brain injury event increases the risk of dementia, brain-age estimation could provide a novel and efficient indexing of the long-term consequences of TBI. Brain-age procedures use predictive modeling to calculate brain-age scores for an individual using structural magnetic resonance imaging (MRI) data. Complicated mild, moderate, and severe TBI (cmsTBI) is associated with a higher predicted age difference (PAD), but the progression of PAD over time remains unclear. We sought to examine whether PAD increases as a function of time since injury (TSI) and if injury severity and sex interacted to influence this progression. METHODS: Through the ENIGMA Adult Moderate and Severe (AMS)-TBI working group, we examine the largest TBI sample to date (n = 343), along with controls, for a total sample size of n = 540, to replicate and extend prior findings in the study of TBI brain age. Cross-sectional T1w-MRI data were aggregated across 7 cohorts, and brain age was established using a similar brain age algorithm to prior work in TBI. RESULTS: Findings show that PAD widens with longer TSI, and there was evidence for differences between sexes in PAD, with men showing more advanced brain age. We did not find strong evidence supporting a link between PAD and cognitive performance. INTERPRETATION: This work provides evidence that changes in brain structure after cmsTBI are dynamic, with an initial period of change, followed by relative stability in brain morphometry, eventually leading to further changes in the decades after a single cmsTBI. ANN NEUROL 2024;96:365-377.

Early diagnosis and treatment of Alzheimer's disease by targeting toxic soluble Aß oligomers.

Transient soluble oligomers of amyloid-ß (Aß) are toxic and accumulate early prior to insoluble plaque formation and cognitive impairment in Alzheimer's disease (AD). Synthetic cyclic D,L-α-peptides (e.g., 1) self-assemble into cross ß-sheet nanotubes, react with early Aß species (1-3 mers), and inhibit Aß aggregation and toxicity in stoichiometric concentrations, in vitro. Employing a semicarbazide as an aza-glycine residue with an extra hydrogen-bond donor to tune nanotube assembly and amyloid engagement, [azaGly6]-1 inhibited Aß aggregation and toxicity at substoichiometric concentrations. High-resolution NMR studies revealed dynamic interactions between [azaGly6]-1 and Aß42 residues F19 and F20, which are pivotal for early dimerization and aggregation. In an AD mouse model, brain positron emission tomography (PET) imaging using stable 64Cu-labeled (aza)peptide tracers gave unprecedented early amyloid detection in 44-d presymptomatic animals. No tracer accumulation was detected in the cortex and hippocampus of 44-d-old 5xFAD mice; instead, intense PET signal was observed in the thalamus, from where Aß oligomers may spread to other brain parts with disease progression. Compared with standard 11C-labeled Pittsburgh compound-B (11C-PIB), which binds specifically fibrillar Aß plaques, 64Cu-labeled (aza)peptide gave superior contrast and uptake in young mouse brain correlating with Aß oligomer levels. Effectively crossing the blood-brain barrier (BBB), peptide 1 and [azaGly6]-1 reduced Aß oligomer levels, prolonged lifespan of AD transgenic Caenorhabditis elegans, and abated memory and behavioral deficits in nematode and murine AD models. Cyclic (aza)peptides offer novel promise for early AD diagnosis and therapy.

Niche conservatism and spread explain introgression between native and invasive fish.

Hybridisation can be an important driver of evolutionary change, but hybridisation with invasive species can have adverse effects on native biodiversity. While hybridisation has been documented across taxa, there is limited understanding of ecological factors promoting patterns of hybridisation and the spatial distribution of hybrid individuals. We combined the results of ecological niche modelling (ENM) and restriction site-associated DNA sequencing to test theories of niche conservatism and biotic resistance on the success of invasion, admixture, and extent of introgression between native and non-native fishes. We related Maxent predictions of habitat suitability based on the native ranges of invasive Eastern Banded Killifish (Fundulus diaphanus diaphanus Lesueur 1817) and native Western Banded Killifish (Fundulus diaphanus menona Jordan and Copeland 1877) to admixture indices of individual Banded Killifish. We found that Eastern Banded Killifish predominated at sites predicted as suitable from their ENM, consistent with niche conservatism. Admixed individuals were more common as Eastern Banded Killifish habitat suitability declined. We also found that Eastern Banded Killifish were most common at sites closest to the presumed source of this invasion, whereas the proportion of admixed individuals increased with distance from the source of invasion. Lastly, we found little evidence that habitat suitability for Western Banded Killifish provides biotic resistance from either displacement by, or admixture with, invasive Eastern Banded Killifish. Our study demonstrates that ENMs can inform conservation-relevant outcomes between native and invasive taxa while emphasising the importance of protecting isolated Western Banded Killifish populations from invasive conspecifics.

Ten simple rules for using public biological data for your research.

With an increasing amount of biological data available publicly, there is a need for a guide on how to successfully download and use this data. The 10 simple rules for using public biological data are: (1) use public data purposefully in your research; (2) evaluate data for your use case; (3) check data reuse requirements and embargoes; (4) be aware of ethics for data reuse; (5) plan for data storage and compute requirements; (6) know what you are downloading; (7) download programmatically and verify integrity; (8) properly cite data; (9) make reprocessed data and models Findable, Accessible, Interoperable, and Reusable (FAIR) and share; and (10) make pipelines and code FAIR and share. These rules are intended as a guide for researchers wanting to make use of available data and to increase data reuse and reproducibility.

Rate of abnormalities in quantitative MR neuroimaging of persons with chronic traumatic brain injury.

BACKGROUND: Mild traumatic brain injury (mTBI) can result in lasting brain damage that is often too subtle to detect by qualitative visual inspection on conventional MR imaging. Although a number of FDA-cleared MR neuroimaging tools have demonstrated changes associated with mTBI, they are still under-utilized in clinical practice. METHODS: We investigated a group of 65 individuals with predominantly mTBI (60 mTBI, 48 due to motor-vehicle collision, mean age 47 ± 13 years, 27 men and 38 women) with MR neuroimaging performed in a median of 37 months post-injury. We evaluated abnormalities in brain volumetry including analysis of left-right asymmetry by quantitative volumetric analysis, cerebral perfusion by pseudo-continuous arterial spin labeling (PCASL), white matter microstructure by diffusion tensor imaging (DTI), and neurometabolites via magnetic resonance spectroscopy (MRS). RESULTS: All participants demonstrated atrophy in at least one lobar structure or increased lateral ventricular volume. The globus pallidi and cerebellar grey matter were most likely to demonstrate atrophy and asymmetry. Perfusion imaging revealed significant reductions of cerebral blood flow in both occipital and right frontoparietal regions. Diffusion abnormalities were relatively less common though a subset analysis of participants with higher resolution DTI demonstrated additional abnormalities. All participants showed abnormal levels on at least one brain metabolite, most commonly in choline and N-acetylaspartate. CONCLUSION: We demonstrate the presence of coup-contrecoup perfusion injury patterns, widespread atrophy, regional brain volume asymmetry, and metabolic aberrations as sensitive markers of chronic mTBI sequelae. Our findings expand the historic focus on quantitative imaging of mTBI with DTI by highlighting the complementary importance of volumetry, arterial spin labeling perfusion and magnetic resonance spectroscopy neurometabolite analyses in the evaluation of chronic mTBI.

The possible clinical utility of the alcohol biomarker phosphatidylethanol for managing suspected alcohol withdrawal in hospitalized patients: A case report.

BACKGROUND AND OBJECTIVES: The decision to initiate pharmacotherapy for alcohol withdrawal is typically based on examining self-reported use of alcohol and symptoms of withdrawal. Phosphatidylethanol (PEth) is a biomarker that could aim in clinical decision-making in withdrawal management. METHODS: This report describes three cases highlighting the potential clinical utility of PEth in caring for individuals at risk for alcohol withdrawal. RESULTS: Two of the cases received phenobarbital when their PEth showed that the risk of withdrawal was low and one case where PEth could have shown this was needed. The results were only available in a delayed fashion, however, could have been useful in informing clinical care. DISCUSSION AND CONCLUSION: PEth can be a useful tool if available without delay. PEth can be used to quickly rule out alcohol withdrawal and avoid misdiagnoses and prolonged hospital stays. SCIENTIFIC SIGNIFICANCE: This is a clinical case study available looking at PEth and withdrawal in hospitalized patients. It proposes that PEth can be used as a way to quickly rule out alcohol withdrawal to avoid misdiagnoses and the possibility of a prolonged hospital stay.

Computer-Assisted Arthroscopic Cam Resection.

Determining the appropriate femoral cam resection during hip arthroscopy for femoroacetabular impingement syndrome is both critical for the patient and challenging for the surgeon. Incomplete bone resection is a leading cause of failed hip arthroscopy, whereas over-resection may increase the risk of femoral neck fracture. The alpha angle is a validated 2-dimensional radiographic measurement used to both diagnose femoroacetabular impingement syndrome preoperatively and to determine resection adequacy postoperatively. Computer-assisted intraoperative guidance systems enhance the accuracy of femoral cam resection, although a preoperative 3-dimensional computed tomography scan may be required. Other systems, such as the HipCheck software (Stryker, Kalamazoo, MI), have been developed to provide intraoperative guidance with live feedback using simultaneous alpha angle measurements overlayed on fluoroscopic images without the requirement for routine preoperative computed tomography. Via intraoperative touchscreen navigation, the surgeon identifies the midpoint of the femoral neck and femoral head. A commercial software program provides real-time alpha angle measurements, as well as enhanced visualization of the femoral cam deformity with an adjustable resection curve. Before the surgeon performs the cam resection, the software provides a template for appropriate resection depth in 6 positions of the hip. Upon completion of the femoral cam resection, the hip is again assessed in the same 6 positions and the alpha angle is assessed to ensure a complete resection.

Cartilage Defects Are Negatively Associated With Long-Term Hip Survivorship Following Contemporary Hip Arthroscopy for Femoroacetabular Impingement Syndrome: A Propensity-Matched Analysis at Minimum 10-Year Follow-Up.

PURPOSE: To identify the timing and risk factors associated with secondary surgery following primary hip arthroscopy for femoroacetabular impingement syndrome (FAIS) at 10-year minimum follow-up. METHODS: A prospectively collected clinical repository was evaluated for cases of primary hip arthroscopy for FAIS between January 2012 and February 2013 with minimum 10-year follow-up. Patients who underwent secondary surgery were propensity matched 1:4 to patients who did not undergo secondary surgery, controlling for age, sex, and body mass index (BMI). The groups were compared on demographics, radiographs, intraoperative findings, operative procedures, and patient-reported outcomes. A Kaplan-Meier survivorship curve was generated. Among the reoperation-free survivors, minimal clinically important difference (MCID) and patient acceptable symptom state (PASS) achievement were recorded for Hip Outcome Score-Activities of Daily Living (HOS-ADL), Hip Outcome Score-Sports Specific (HOS-SS) subscales, modified Harris Hip Score (mHHS), 12-item international Hip Outcome Tool (iHOT-12), and Visual Analog Scale for Pain (VAS Pain). RESULTS: Twenty-four reoperation patients (67% female; age 40.1 ± 14.3 years; BMI 27.2 ± 5.5) were matched to 96 reoperation-free patients (62% female; age 37.0 ± 10.8 years; BMI 25.2 ± 4.7, P ≥ .111). Mean follow-up was 10.3 ± 0.2 years. No preoperative demographic differences were found between groups. The reoperation group showed more high-grade cartilage defects on the acetabulum and femoral head (33% vs 8%, P = .004; 29% vs 7%, P = .007). A bimodal distribution of time to reoperation was evidenced independent of the secondary surgery performed. Among the reoperation-free survivors, MCID and PASS achievement was as follows: HOS-ADL (69.1%, 62.1%), HOS-SS (69.9%, 74.4%), mHHS (73.3%, 58.1%), iHOT-12 (n/a, 63.8%), and VAS-Pain (80.2%, 62.6%). CONCLUSIONS: Patients requiring reoperation following primary hip arthroscopy for FAIS demonstrated more severe cartilage defects and a bimodal distribution of time to reoperation. LEVEL OF EVIDENCE: Level III, retrospective comparative case series.

Six-Month Outcomes Correlate With 10-Year Outcomes After Hip Arthroscopy for Femoroacetabular Impingement Syndrome.

PURPOSE: To identify whether 6-month outcomes after hip arthroscopy for femoroacetabular impingement syndrome (FAIS) correlate with outcomes at minimum 10-year follow-up. METHODS: Patients who underwent primary hip arthroscopy for FAIS from 2012 to 2013 were reviewed and included if they had 6-month and minimum 10-year follow-up. Patient-reported outcome (PRO) measures included the Hip Outcome Score Activities of Daily Living (HOS-ADL) subscale, Hip Outcome Score Sports-Specific (HOS-SS) subscale, modified Harris Hip Score (mHHS), visual analog scale (VAS) for pain, and VAS for satisfaction. We compared 6-month and 10-year outcome scores and analyzed the relations between 6-month and 1-, 2-, 5-, and 10-year outcome scores using Pearson correlation coefficients (r). Six-month scores and clinically significant outcome achievement were then compared with 10-year clinically significant outcome achievement and reoperations, including revision hip arthroscopy and conversion to total hip arthroplasty (THA), using logistic regressions and the Fisher exact test. RESULTS: This study included 60 patients (60.0% female sex; mean age, 36.0 ± 12.2 years). The mHHS, VAS pain score, and VAS satisfaction score significantly improved from 6-month to 10-year follow-up (P ≤ .021), whereas the HOS-ADL and HOS-SS did not (P ≥ .072). There were significant correlations between 6-month and 10-year scores for the HOS-ADL (r = 0.505), HOS-SS (r = 0.592), and mHHS (r = 0.362) (P ≤ .022 for all), as well as significant correlations between 6-month and 1-, 2-, and 5-year scores (P ≤ .014 for all). The 6-month HOS-ADL, HOS-SS, and mHHS were all significantly associated with their respective 10-year achievement of the patient acceptable symptom state (PASS) (P ≤ .044). Furthermore, 6-month HOS-ADL and mHHS were significantly associated with THA conversion (P ≤ .041). Comparable 6-month and 10-year achievement of the minimal clinically important difference (96.5% vs 97.8%, P > .999) and PASS (85.2% vs 87.5%, P > .999) for any PRO was observed. CONCLUSIONS: After hip arthroscopy for FAIS, patients' 6-month HOS-ADL and mHHS were significantly associated with their 10-year PROs, PASS achievement, and THA conversion, although correlation strengths decreased with increasing time from surgery. LEVEL OF EVIDENCE: Level IV, case series.

Patients Aged 40 Years and Older Demonstrate Durable and Comparable Results to Patients Aged Less Than 40 Years After Primary Hip Arthroscopy for Femoroacetabular Impingement Syndrome: A Propensity Matched Study at Minimum 10-Year Follow-Up.

PURPOSE: The purpose of the present study was to compare clinical outcomes and rates of secondary surgery, including revision hip arthroscopy and conversion to total hip arthroplasty (THA), after primary hip arthroscopy for femoroacetabular impingement syndrome (FAIS) in patients ≥40 years of age at minimum 10-year follow-up compared with a propensity-matched control group of patients <40 years. METHODS: A retrospective cohort study was performed for patients who underwent primary hip arthroscopy for FAIS between January 2012 and February 2013. Patients ≥40 years old were propensity matched in a 1:1 ratio by sex and body mass index to patients <40 years old. Patient-reported outcomes (PROs) including Hip Outcome Score for Activities of Daily Living and Sports-Specific subscales, modified Harris Hip, International Hip Outcome Tool-12, and Visual Analog Scale for Pain and Satisfaction were collected. Rates of minimal clinically important difference (MCID) and patient-acceptable symptomatic state (PASS) achievement at 10 years were evaluated and compared between groups. Rates of secondary surgery including revision hip arthroscopy and conversion to THA were evaluated. Gross survivorship between cohorts was evaluated using a Kaplan-Meier curve. RESULTS: Fifty-three patients aged ≥40 (age 48.3 ± 5.8 years) were successfully matched to 53 patients aged <40 (age: 28.9 ± 7.2, <0.001). There were no other preoperative group differences regarding patient demographics, characteristics, or radiographic findings. Both groups demonstrated significant improvement regarding all PROs at a minimum of 10 years' follow-up (P < .001 for all). No significant difference was noted between cohorts regarding any delta (preoperative to 10-year postoperative) scores (P > .05 for all). High rates of MCID and PASS achievement were achieved in both cohorts, with no significant differences in any PRO measure (P > .05 for all). No significant differences in rates of complications (age ≥40: 2.0%, age <40: 7.7%, P = .363), rates of revision (age ≥40: 7.5%, age <40: 9.4%, P = .999), or conversion to THA (age ≥40: 13.2%, age <40: 3.8%, P = .161) were identified. On Kaplan-Meier analysis, no significant difference (P = .321) was demonstrated in overall gross survivorship between cohorts. CONCLUSION: Patients with age ≥40 with FAIS undergoing primary hip arthroscopy demonstrated durable and comparable 10-year PRO and rates of MCID and PASS achievement compared with a propensity-matched cohort of age <40 counterparts. LEVEL OF EVIDENCE: Level III, retrospective comparative prognostic trial.

Females and Males Achieve Similar Improvement, Outcomes, and Survivorship Following Hip Arthroscopy with Labral and Capsular Repair for Femoroacetabular Impingement Syndrome at Minimum 10-Year Follow-Up.

PURPOSE: To evaluate the effect of patient sex on 10-year patient-reported outcomes (PROs) and survivorship after hip arthroscopy (HA) for femoroacetabular impingement syndrome (FAIS). METHODS: Patients who underwent primary HA for FAIS with minimum 10-year follow-up from 1/2012-12/2013 were retrospectively reviewed. Female patients were propensity-matched to male patients in a 1:1 ratio by age and body mass index. PROs and rates of minimal clinically important difference (MCID) and patient-acceptable symptom state (PASS) achievement were compared between cohorts. Rate of reoperation-free survivorship was compared between sexes. RESULTS: One-hundred and twenty-one- females (age: 36.2 ± 12.3 years) were matched to 121 males (age: 35.7 ± 11.3 years, p = 0.594) at average follow-up of 10.4 ± 0.4 years. There were no differences in any preoperative demographic characteristics between the groups (p ≥ 0.187). Both groups demonstrated significant improvement in every PRO measure between the preoperative and 10-year postoperative time points (p < 0.001). The magnitude of improvement was similar between the groups for all PRO measures (p ≥ 0.139). At 10-years, female patients trended towards higher MCID achievement for the Hip Outcome Score-Activities of Daily Living subscale (HOS-ADL) than male patients (72.7% vs. 57.3%, p = 0.061), with otherwise similar MCID achievement rates. Females trended towards significantly lower HOS-Sports Subscale PASS achievement (65.4% vs. 77.1%, p = 0.121) with otherwise similar PASS achievement rates between the groups (p ≥ 0.170). CONCLUSION: Female and male patients experienced similar improvement in PROs at ten-year follow-up. MCID and PASS achievement rates were predominantly similar between sexes. Survivorship did not differ between groups. Long-term success can be expected for appropriately indicated patients undergoing HA for FAIS, regardless of sex. LEVEL OF EVIDENCE: III, Retrospective Cohort Study.

Patients With Lateral and Anterolateral Cam Morphology Have Greater Deformities Versus Typical Anterolateral Deformity Alone but No Differences in Postoperative Outcomes: A Propensity-Matched Analysis at Minimum 5-Year Follow-Up.

PURPOSE: To compare pre- and postoperative findings between patients undergoing hip arthroscopy for femoroacetabular impingement syndrome with lateral impingement versus those without lateral impingement METHODS: Patients who underwent primary hip arthroscopy for femoroacetabular impingement syndrome between 2012 and 2017 with minimum 5-year follow-up were included. Alpha angle (AA) was measured on preoperative anteroposterior (AP) and 90° Dunn radiographs. Patients with AA >60° on Dunn view but not AP view (no lateral impingement) were propensity matched by sex, age, and body mass index in a 1:3 ratio to patients with AA >60° on both views (lateral impingement). Demographic characteristics, radiographic and intraoperative findings, reoperation rates, and patient-reported outcomes (PROs) were compared between groups. Categorical variables were compared using the Fisher exact testing and continuous variable using 2-tailed Student t tests. RESULTS: Sixty patients with lateral impingement (65.0% female, age: 35.3 ± 13.0 years) were matched to 180 patients without lateral impingement (65.0% female, age: 34.7 ± 12.5 years, P ≥ .279). Patients with lateral impingement had larger preoperative AAs on both Dunn (71.0° ± 8.8° vs 67.6° ± 6.1°, P = .001) and AP radiographs (79.0° ± 12.1° vs 48.2° ± 6.5°, P < .001). However, there were no differences in postoperative AAs on either view (Dunn: 39.0° ± 6.1° vs 40.5° ± 5.3°, AP: 45.8° ± 9.0° vs 44.9° ± 7.0°, P ≥ .074). Labral tears began more superiorly in patients with lateral impingement (12:00 ± 0:49 vs 12:17 ± 0:41, P = .030), and they demonstrated greater rates of acetabular and femoral cartilage damage (P = .030 for both); however, there were no differences in PROs or reoperation rates between the groups at 5-year follow-up. CONCLUSIONS: Although cam deformities located laterally and anterolaterally are larger than those located anterolaterally alone, both can be resected adequately, resulting in similar postoperative radiographic measurements, PROs, and survivorship. LEVEL OF EVIDENCE: Level III, retrospective cohort study.

Enhancing diagnostic precision for acute chest syndrome in sickle cell disease: insights from dual-energy CT lung perfusion mapping.

PURPOSE: Acute chest syndrome (ACS) is secondary to occlusion of the pulmonary vasculature and a potentially life-threatening complication of sickle cell disease (SCD). Dual-energy CT (DECT) iodine perfusion map reconstructions can provide a method to visualize and quantify the extent of pulmonary microthrombi. METHODS: A total of 102 patients with sickle cell disease who underwent DECT CTPA with perfusion were retrospectively identified. The presence or absence of airspace opacities, segmental perfusion defects, and acute or chronic pulmonary emboli was noted. The number of segmental perfusion defects between patients with and without acute chest syndrome was compared. Sub-analyses were performed to investigate robustness. RESULTS: Of the 102 patients, 68 were clinically determined to not have ACS and 34 were determined to have ACS by clinical criteria. Of the patients with ACS, 82.4% were found to have perfusion defects with a median of 2 perfusion defects per patient. The presence of any or new perfusion defects was significantly associated with the diagnosis of ACS (P = 0.005 and < 0.001, respectively). Excluding patients with pulmonary embolism, 79% of patients with ACS had old or new perfusion defects, and the specificity for new perfusion defects was 87%, higher than consolidation/ground glass opacities (80%). CONCLUSION: DECT iodine map has the capability to depict microthrombi as perfusion defects. The presence of segmental perfusion defects on dual-energy CT maps was found to be associated with ACS with potential for improved specificity and reclassification.

Optimisation of virtual monoenergetic reconstructions for the diagnosis of pulmonary embolism using photon-counting detector computed tomography angiography.

Purpose: Computed tomography (CT) pulmonary angiography is considered the gold standard for pulmonary embolism (PE) diagnosis, relying on the discrimination between contrast and embolus. Photon-counting detector CT (PCD-CT) generates monoenergetic reconstructions through energy-resolved detection. Virtual monoenergetic images (VMI) at low keV can be used to improve pulmonary artery opacification. While studies have assessed VMI for PE diagnosis on dual-energy CT (DECT), there is a lack of literature on optimal settings for PCD-CT-PE reconstructions, warranting further investigation. Material and methods: Twenty-five sequential patients who underwent PCD-CT pulmonary angiography for suspicion of acute PE were retrospectively included in this study. Quantitative metrics including signal-to-noise ratio (SNR) and contrast-to-noise (CNR) ratio were calculated for 4 VMI values (40, 60, 80, and 100 keV). Qualitative measures of diagnostic quality were obtained for proximal to distal pulmonary artery branches by 2 cardiothoracic radiologists using a 5-point modified Likert scale. Results: SNR and CNR were highest for the 40 keV VMI (49.3 ± 22.2 and 48.2 ± 22.1, respectively) and were inversely related to monoenergetic keV. Qualitatively, 40 and 60 keV both exhibited excellent diagnostic quality (mean main pulmonary artery: 5.0 ± 0 and 5.0 ± 0; subsegmental pulmonary arteries 4.9 ± 0.1 and 4.9 ± 0.1, respectively) while distal segments at high (80-100) keVs had worse quality. Conclusions: 40 keV was the best individual VMI for the detection of pulmonary embolism by quantitative metrics. Qualitatively, 40-60 keV reconstructions may be used without a significant decrease in subjective quality. VMIs at higher keV lead to reduced opacification of the distal pulmonary arteries, resulting in decreased image quality.

Cell-type-specific gene expression and regulation in the cerebral cortex and kidney of atypical Setbp1S858R Schinzel Giedion Syndrome mice.

Schinzel Giedion Syndrome (SGS) is an ultra-rare autosomal dominant Mendelian disease presenting with abnormalities spanning multiple organ systems. The most notable phenotypes involve severe developmental delay, progressive brain atrophy, and drug-resistant seizures. SGS is caused by spontaneous variants in SETBP1, which encodes for the epigenetic hub SETBP1 transcription factor (TF). SETBP1 variants causing classical SGS cluster at the degron, disrupting SETBP1 protein degradation and resulting in toxic accumulation, while those located outside cause milder atypical SGS. Due to the multisystem phenotype, we evaluated gene expression and regulatory programs altered in atypical SGS by snRNA-seq of the cerebral cortex and kidney of Setbp1S858R heterozygous mice (corresponds to the human likely pathogenic SETBP1S867R variant) compared to matched wild-type mice by constructing cell-type-specific regulatory networks. Setbp1 was differentially expressed in excitatory neurons, but known SETBP1 targets were differentially expressed and regulated in many cell types. Our findings suggest molecular drivers underlying neurodevelopmental phenotypes in classical SGS also drive atypical SGS, persist after birth, and are present in the kidney. Our results indicate SETBP1's role as an epigenetic hub leads to cell-type-specific differences in TF activity, gene targeting, and regulatory rewiring. This research provides a framework for investigating cell-type-specific variant impact on gene expression and regulation.

Structural insights into the role of the WW2 domain on tandem WW-PPxY motif interactions of oxidoreductase WWOX.

Class I WW domains are present in many proteins of various functions and mediate protein interactions by binding to short linear PPxY motifs. Tandem WW domains often bind peptides with multiple PPxY motifs, but the interplay of WW-peptide interactions is not always intuitive. The WW domain-containing oxidoreductase (WWOX) harbors two WW domains: an unstable WW1 capable of PPxY binding and stable WW2 that cannot bind PPxY. The WW2 domain has been suggested to act as a WW1 domain chaperone, but the underlying mechanism of its chaperone activity remains to be revealed. Here, we combined NMR, isothermal calorimetry, and structural modeling to elucidate the roles of both WW domains in WWOX binding to its PPxY-containing substrate ErbB4. Using NMR, we identified an interaction surface between these two domains that supports a WWOX conformation compatible with peptide substrate binding. Isothermal calorimetry and NMR measurements also indicated that while binding affinity to a single PPxY motif is marginally increased in the presence of WW2, affinity to a dual-motif peptide increases 10-fold. Furthermore, we found WW2 can directly bind double-motif peptides using its canonical binding site. Finally, differential binding of peptides in mutagenesis experiments was consistent with a parallel N- to C-terminal PPxY tandem motif orientation in binding to the WW1-WW2 tandem domain, validating structural models of the interaction. Taken together, our results reveal the complex nature of tandem WW-domain organization and substrate binding, highlighting the contribution of WWOX WW2 to both protein stability and target binding.

An Antigenically Diverse, Representative Panel of Envelope Glycoproteins for Hepatitis C Virus Vaccine Development.

BACKGROUND & AIMS: Development of a prophylactic hepatitis C virus (HCV) vaccine will require accurate and reproducible measurement of neutralizing breadth of vaccine-induced antibodies. Currently available HCV panels may not adequately represent the genetic and antigenic diversity of circulating HCV strains, and the lack of standardization of these panels makes it difficult to compare neutralization results obtained in different studies. Here, we describe the selection and validation of a genetically and antigenically diverse reference panel of 15 HCV pseudoparticles (HCVpps) for neutralization assays. METHODS: We chose 75 envelope (E1E2) clones to maximize representation of natural polymorphisms observed in circulating HCV isolates, and 65 of these clones generated functional HCVpps. Neutralization sensitivity of these HCVpps varied widely. HCVpps clustered into 15 distinct groups based on patterns of relative sensitivity to 7 broadly neutralizing monoclonal antibodies. We used these data to select a final panel of 15 antigenically representative HCVpps. RESULTS: Both the 65 and 15 HCVpp panels span 4 tiers of neutralization sensitivity, and neutralizing breadth measurements for 7 broadly neutralizing monoclonal antibodies were nearly equivalent using either panel. Differences in neutralization sensitivity between HCVpps were independent of genetic distances between E1E2 clones. CONCLUSIONS: Neutralizing breadth of HCV antibodies should be defined using viruses spanning multiple tiers of neutralization sensitivity rather than panels selected solely for genetic diversity. We propose that this multitier reference panel could be adopted as a standard for the measurement of neutralizing antibody potency and breadth, facilitating meaningful comparisons of neutralization results from vaccine studies in different laboratories.