Ultraviolet Photodissociation Dynamics of the 1-Methylallyl Radical.

The ultraviolet (UV) photodissociation dynamics of the 1-methylallyl (1-MA) radical were studied using the high-n Rydberg atom time-of-flight (HRTOF) technique in the wavelength region of 226-244 nm. The 1-MA radicals were produced by 193 nm photodissociation of the 3-chloro-1-butene and 1-chloro-2-butene precursor. The 1 + 1 REMPI spectrum of 1-MA agrees with the previous UV absorption spectrum in this wavelength region. Quantum chemistry calculations show that the UV absorption is mainly attributed to the 3pz Rydberg state (perpendicular to the allyl plane). The H atom photofragment yield (PFY) spectrum of 1-MA from 3-chloro-1-butene displays a broad peak around 230 nm, while that from 1-chloro-2-butene peaks at ∼236 nm. The translational energy distributions of the H atom loss product channel, P (ET)'s, show a bimodal distribution indicating two dissociation pathways in 1-MA. The major pathway is isotropic in product angular distribution with ß âˆ¼ 0 and has a low fraction of average translational energy in the total excess energy, ⟨fT⟩, in the range of 0.13-0.17; this pathway corresponds to unimolecular dissociation of 1-MA after internal conversion to form 1,3-butadiene + H. The minor pathway is anisotropic with ß âˆ¼ -0.23 and has a large ⟨fT⟩ of ∼0.62-0.72. This fast pathway suggests a direct dissociation of the methyl H atom on a repulsive excited state surface or the repulsive part of the ground state surface to form 1,3-butadiene + H. The fast/slow pathway branching ratio is in the range of 0.03-0.08.

TDP43 autoregulation gives rise to shortened isoforms that are tightly controlled by both transcriptional and post-translational mechanisms.

The nuclear RNA-binding protein TDP43 is integrally involved in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Previous studies uncovered N-terminal TDP43 isoforms that are predominantly cytosolic in localization, highly prone to aggregation, and enriched in susceptible spinal motor neurons. In healthy cells, however, these shortened (s)TDP43 isoforms are difficult to detect in comparison to full-length (fl)TDP43, raising questions regarding their origin and selective regulation. Here, we show that sTDP43 is created as a byproduct of TDP43 autoregulation and cleared by nonsense mediated RNA decay (NMD). The sTDP43-encoding transcripts that escape NMD can lead to toxicity but are rapidly degraded post-translationally. Circumventing these regulatory mechanisms by overexpressing sTDP43 results in neurodegeneration in vitro and in vivo via N-terminal oligomerization and impairment of flTDP43 splicing activity, in addition to RNA binding-dependent gain-of-function toxicity. Collectively, these studies highlight endogenous mechanisms that tightly regulate sTDP43 expression and provide insight into the consequences of aberrant sTDP43 accumulation in disease.

Bispecific antibody shuttles targeting CD98hc mediate efficient and long-lived brain delivery of IgGs.

The inability of antibodies to penetrate the blood-brain barrier (BBB) is a key limitation to their use in diverse applications. One promising strategy is to deliver IgGs using a bispecific BBB shuttle, which involves fusing an IgG to a second affinity ligand that engages a cerebrovascular endothelial target and facilitates transport across the BBB. Nearly all prior efforts have focused on shuttles that target transferrin receptor (TfR-1) despite inherent delivery and safety challenges. Here, we report bispecific antibody shuttles that engage CD98hc, the heavy chain of the large neutral amino acid transporter (LAT1), and efficiently transport IgGs into the brain. Notably, CD98hc shuttles lead to much longer-lived brain retention of IgGs than TfR-1 shuttles while enabling more specific targeting due to limited CD98hc engagement in the brain parenchyma, which we demonstrate for IgGs that either agonize a neuronal receptor (TrkB) or target other endogenous cell-surface proteins on neurons and astrocytes.

Quantitative flow cytometric selection of tau conformational nanobodies specific for pathological aggregates.

Single-domain antibodies, also known as nanobodies, are broadly important for studying the structure and conformational states of several classes of proteins, including membrane proteins, enzymes, and amyloidogenic proteins. Conformational nanobodies specific for aggregated conformations of amyloidogenic proteins are particularly needed to better target and study aggregates associated with a growing class of associated diseases, especially neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. However, there are few reported nanobodies with both conformational and sequence specificity for amyloid aggregates, especially for large and complex proteins such as the tau protein associated with Alzheimer's disease, due to difficulties in selecting nanobodies that bind to complex aggregated proteins. Here, we report the selection of conformational nanobodies that selectively recognize aggregated (fibrillar) tau relative to soluble (monomeric) tau. Notably, we demonstrate that these nanobodies can be directly isolated from immune libraries using quantitative flow cytometric sorting of yeast-displayed libraries against tau aggregates conjugated to quantum dots, and this process eliminates the need for secondary nanobody screening. The isolated nanobodies demonstrate conformational specificity for tau aggregates in brain samples from both a transgenic mouse model and human tauopathies. We expect that our facile approach will be broadly useful for isolating conformational nanobodies against diverse amyloid aggregates and other complex antigens.

Toward structuring real-world data: Deep learning for extracting oncology information from clinical text with patient-level supervision.

Most detailed patient information in real-world data (RWD) is only consistently available in free-text clinical documents. Manual curation is expensive and time consuming. Developing natural language processing (NLP) methods for structuring RWD is thus essential for scaling real-world evidence generation. We propose leveraging patient-level supervision from medical registries, which are often readily available and capture key patient information, for general RWD applications. We conduct an extensive study on 135,107 patients from the cancer registry of a large integrated delivery network (IDN) comprising healthcare systems in five western US states. Our deep-learning methods attain test area under the receiver operating characteristic curve (AUROC) values of 94%-99% for key tumor attributes and comparable performance on held-out data from separate health systems and states. Ablation results demonstrate the superiority of these advanced deep-learning methods. Error analysis shows that our NLP system sometimes even corrects errors in registrar labels.

Bispecific antibody shuttles targeting CD98hc mediate efficient and long-lived brain delivery of IgGs.

The inability of antibodies and other biologics to penetrate the blood-brain barrier (BBB) is a key limitation to their use in diagnostic, imaging, and therapeutic applications. One promising strategy is to deliver IgGs using a bispecific BBB shuttle, which involves fusing an IgG with a second affinity ligand that engages a cerebrovascular endothelial target and facilitates transport across the BBB. Nearly all prior efforts have focused on the transferrin receptor (TfR-1) as the prototypical endothelial target despite inherent delivery and safety challenges. Here we report bispecific antibody shuttles that engage CD98hc (also known as 4F2 and SLC3A2), the heavy chain of the large neutral amino acid transporter (LAT1), and efficiently transport IgGs into the brain parenchyma. Notably, CD98hc shuttles lead to much longer-lived brain retention of IgGs than TfR-1 shuttles while enabling more specific brain targeting due to limited CD98hc engagement in the brain parenchyma. We demonstrate the broad utility of the CD98hc shuttles by reformatting three existing IgGs as CD98hc bispecific shuttles and delivering them to the mouse brain parenchyma that either agonize a neuronal receptor (TrkB) or target other endogenous antigens on specific types of brain cells (neurons and astrocytes).

Immunopathological Alterations after Blast Injury and Hemorrhage in a Swine Model of Prolonged Damage Control Resuscitation.

Trauma-related hemorrhagic shock (HS) remains a leading cause of death among military and civilian trauma patients. We have previously shown that administration of complement and HMGB1 inhibitors attenuate morbidity and mortality 24 h after injury in a rat model of blast injury (BI) and HS. To further validate these results, this study aimed to develop a swine model and evaluate BI+HS-induced pathophysiology. Anesthetized Yucatan minipigs underwent combined BI and volume-controlled hemorrhage. After 30 min of shock, animals received an intravenous bolus of PlasmaLyte A and a continuous PlasmaLyte A infusion. The survival rate was 80% (4/5), and the non-survivor expired 72 min post-BI. Circulating organ-functional biomarkers, inflammatory biomarkers, histopathological evaluation, and CT scans indicated evidence of multiple-organ damage, systemic innate immunological activation, and local tissue inflammation in the injured animals. Interestingly, a rapid and dramatic increase in plasma levels of HMGB1 and C3a and markedly early myocarditis and encephalitis were associated with early death post-BI+HS. This study suggests that this model reflects the immunopathological alterations of polytrauma in humans during shock and prolonged damage control resuscitation. This experimental protocol could be helpful in the assessment of immunological damage control resuscitation approaches during the prolonged care of warfighters.

Technique of stent sizing in patients with symptomatic chronic iliofemoral venous obstruction-the case for intravascular ultrasound-determined inflow channel luminal area-based stenting and associated long-term outcomes.

OBJECTIVE: Femoroiliocaval stenting has become the standard of care for patients with quality-of-life impairing chronic iliofemoral venous obstruction not responding to conservative measures. Although improvement after stenting has been noted in multiple large studies, sizing of stents has been subjective in nature with a general tendency to use smaller stents that would be required to relieve venous hypertension. This study evaluates the authors' technique of using the intravascular ultrasound (IVUS) inflow channel luminal area to guide stent sizing. METHODS: Patients who underwent femoroiliocaval stenting for quality-of-life impairing chronic iliofemoral venous obstruction and had failed conservative therapy from 2015 to 2021 were included in the study. Clinical outcomes including venous clinical severity score (VCSS), visual analog scale (VAS) pain score, and grade of swelling (GOS) were appraised before and after stenting. Also evaluated were quality of life (Chronic Venous Insufficiency Questionnaire-20 [CIVIQ-20] instrument) and stent outcomes including patencies and reinterventions. Comparisons were made between limbs that underwent placement of larger caliber stents (largest stent diameter >20 mm: >20 mm stent group) vs smaller caliber stents (largest stent diameter ≤20 mm: ≤20 mm stent group). t tests and analysis of variance were used to compare outcomes, whereas the Kaplan-Meier analysis was used to evaluate patencies with log rank used to compare the curves. RESULTS: A total of 300 patients (300 limbs) underwent stenting with a median age of 58 years. There was a preponderance of men (159 of 300), left laterality (176 of 300), and post-thrombotic syndrome (176 of 300). The median body mass index was 41. There were 120 limbs in the >20 mm stent group and 180 limbs in the ≤20 mm stent group. The median follow-up was 23 months. There was no significant difference in baseline VCSS, VAS pain score, or GOS between the two groups. However, there was a significant difference in IVUS-determined inflow channel luminal area between the two groups (228 mm2 >20 mm stent group vs 176 mm2 for ≤20 mm stent group [P < .0001]). After stenting there was a significant improvement in the VCSS, VAS pain score, and GOS at 6 weeks, 3, 6, 12, and 24 months (P < .0001) without any difference between the groups (P > .05). The CIVIQ-20 score also improved from 58 to 38 (P < .0001) for the entire cohort and for the two groups (P < .0001). Overall primary, primary-assisted, and secondary patencies at 60 months were 84%, 100%, and 100%, respectively. Reintervention rate was 10% without any difference between the groups. CONCLUSIONS: Stent sizing using IVUS-determined inflow channel luminal area in patients undergoing stenting for quality-of-life impairing chronic iliofemoral venous obstruction resulted in a significant improvement in the VCSS, VAS pain score, GOS, and quality of life (CIVIQ-20) after stenting. Excellent stent patencies and low reintervention rates were also noted. IVUS-determined inflow channel luminal area represents an objective technique of stent sizing in comparison to the subjective techniques that currently exist.

A Mixed-Methods Investigation Examining Site-Level Variation in Reach Out and Read Implementation.

OBJECTIVE: Reach Out and Read (ROR) is an evidence-based early childhood intervention that has been implemented at scale, yet description of ROR implementation is inconsistent. This study engages implementation science to examine ROR delivery and site-level variation. METHODS: As part of an ongoing clinical trial, we conducted a mixed-methods study in 3 community health centers (CHCs) that serve low-income Latino families. We integrated quantitative parent survey data, qualitative data from monthly key informant interviews with ROR site leaders over 1 year, and in-depth interviews with 18 additional clinicians. At enrollment, parents reported whether they received a children's book, guidance on reading, and modeling from clinicians. We analyzed quantitative data using descriptive statistics, and qualitative data iteratively engaging emergent and a priori codes drawn from the Template for Intervention Description and Replication Checklist. RESULTS: Three hundred Latino parents (mean age: 31; 75% ≤HS education) completed surveys. The mean child age was 8 months. Overall, most parents reported receiving a book (84%) and guidance (73%), but fewer experienced modeling (23%). Components parents received varied across CHCs. Two themes emerged to explain the variation observed: 1) differences in the perceived purpose of shared reading and book delivery aligned with variation in implementation, and 2) site-level barriers affected what components were implemented. CONCLUSION: Because of substantive variation in ROR implementation across sites, systematic descriptions using established frameworks and corresponding measurement to characterize ROR implementation may enhance our understanding of mechanisms underlying ROR's effects, which clinicians and policymakers can use to maximize ROR's impact.

Plethysmographic features of calf pump failure in chronic venous obstruction and reflux.

BACKGROUND: Calf pump failure (CPF) is a common concept in chronic venous disease. Dorsal vein pressures were originally used to define the pathophysiology. More recently, an abnormal ejection fraction (EF) and residual volume fraction (RVF) with air plethysmography (APG) have been substituted for its diagnosis. The relationship between reflux and calf pump function has been studied extensively. Reflux is thought to be the main cause of CPF, although other mechanisms may play a secondary role. Data mining in our dataset revealed that CPF is frequently found in nonrefluxive limbs-an unexpected finding. We analyzed the APG features of CPF in nonrefluxive limbs of a large cohort of patients investigated for chronic venous disease in our clinic. Data from refluxive limbs (control) seen over the same period was included for comparison. Venous obstructive pathology was variably present in both subsets. Iliac vein stent outcome in CPF limbs from both subsets is included. The role of obstruction in CPF is currently unknown. METHODS: Records of 13,234 limbs in 8813 patients evaluated for suspected chronic venous disease over a 22-year period were analyzed. Prestent and poststent data in 406 CPF limbs (129 nonrefluxive; 277 refluxive) that underwent iliac vein stenting to correct associated stenosis are included. This is a single-center retrospective analysis of prospectively collected data. Duplex and APG data were available for included limbs. A RVF of more than 50% was defined as CPF. A reflux time of greater than 1 second elicited with automated cuffs in the erect position was defined as reflux. RESULTS: There were 7780 (59%) limbs with reflux and 5454 (41%) that were nonrefluxive. Supine venous pressure, an index of venous obstruction, was elevated in both subsets. The incidence of CPF was 25% in refluxive limbs and 16% in nonrefluxive limbs totaling 2790 limbs. Venous volume and venous filling index were significantly elevated (P = .0001) in refluxive limbs compared to nonrefluxive limbs. The EF was diminished (<50%) in all CPF limbs except in a small fraction (n = 427 [3%]). Stent correction of iliac vein stenosis corrected CPF, normalizing the RVF in both subsets. CONCLUSIONS: CPF frequently occurs in nonrefluxive limbs with incidence only slightly less than in refluxive limbs. An RVF of more than 50% seems to be a practical definition of a CPF; an EF of less than 50% is associated with a RVF of greater than 50% in 97% of analyzed limbs. Prospective identification of CPF in limbs with chronic venous disease may allow more detailed investigation of its cause (preload, afterload, neuromuscular pathology or joint immobility, etc) and direct more targeted treatment than currently practiced.

Clinical tolerance of untreated reflux after iliac vein stent placement.

BACKGROUND: We have recently demonstrated in a large patient cohort that the prevalence and severity of reflux will improve in most limbs after stenting and that most limbs will not develop new-onset reflux. In the present report, we have focused on the long-term clinical outcomes associated with untreated reflux in the same patient cohort who had undergone iliofemoral venous stenting without correction of residual reflux. METHODS: The clinical outcomes data from 1379 limbs treated with only iliac vein stenting without correction of superficial or deep reflux from 1997 to 2018 were analyzed (23-year follow-up period). Of the 1379 limbs, 632 (46%) had had preexisting reflux before stenting and 747 (54%) had did not. The reflux data (reflux segmental score, air plethysmography, ambulatory venous pressure) for these patients have been previously reported in detail. The subsets were compared perioperatively with each other using the following variables: grade of swelling, visual analog scale for pain score, venous clinical severity score, venous stasis dermatitis, ulceration, and quality of life measures. RESULTS: Both groups demonstrated improvements in the venous clinical severity score, grade of swelling, visual analog scale score, and quality of life. No differences were found in ulcer healing (5% vs 3% for limbs with and without prestent reflux, respectively) and resolution of dermatitis (6% vs 5% for limbs with and without prestent reflux, respectively) between the two groups. Of the 632 limbs with preexisting reflux, 218 (34%) had had axial reflux and 414 had had nonaxial reflux (66%). The clinical outcomes were similar between the two groups. Using a multisegment reflux score, the limbs with prestent reflux (n = 632) were divided into two groups. A segmental score of ≥3 indicated severe reflux and a score of <3 indicated moderate reflux. Of these 632 limbs, 161 (25%) had severe reflux and 471 (75%) had moderate reflux. The two groups demonstrated similar outcomes for most clinical parameters. The post-thrombotic limbs and nonthrombotic limbs also showed similar outcomes. CONCLUSIONS: The long-term follow-up of patients after iliac vein stenting showed that uncorrected reflux is well tolerated by most patients across most clinical measures.

Improvement following restoration of inline flow argues against comprehensive thrombus removal strategies and for selective stenting in acute symptomatic iliofemoral venous thrombosis.

OBJECTIVE: Randomized trials have demonstrated the benefit of thrombus removal strategies in iliofemoral deep venous thrombosis (IFDVT) in providing early symptom relief and decreasing the incidence of post-thrombotic syndrome (PTS), especially severe PTS. However, the impact of quantum of residual thrombus burden (RTB) on PTS as determined by intravascular ultrasound examination and the role of venous stenting in the acute setting have not been evaluated and represent the focus of this study. METHODS: Sixty-nine limbs (65 patients) undergoing thrombus removal for acute symptomatic IFDVT between 2015 and 2021 formed the study cohort. The Venous Clinical Severity Score (VCSS) (range, 0-27) grade of swelling (GOS) (range, 0-4), and visual analog scale (VAS) pain scores (range, 0-10) were evaluated initially and at 6, 12, and 24 months after thrombus removal. Quality of life was appraised using the CIVIQ-20 instrument. The extent of initial and RTB after the intervention was estimated using intravascular ultrasound examination. Grading was done as less than 50% (1), 50% to 99% (2), or 100% (3) of luminal thrombus fill within each segment (common femoral vein, external iliac vein, and common iliac vein) by a blinded rater and then combined to generate a total score. The use of stenting, both concurrent (severe residual stenosis/persistent occlusion) and delayed (quality of life impairing residual or recurrent symptoms), was evaluated. RESULTS: Of the 69 limbs, 53 underwent pharmacomechanical/mechanical thrombectomy (PMT), whereas 16 patients underwent PMT and catheter-directed thrombolysis with restoration of inline flow in all limbs. Post-intervention VCSS improved from 6 to 2 at 24 months (P < .0001). GOS improved from 4 to 0 at 24 months (P < .0001). The VAS pain score went from 5 to 0 at 6 months (P < .0001) and remained at 0 at 12 months (P < .0001), but increased to 3 at 24 months (P = .02). The CIVIQ-20 score improved from 38 to 22 (P = .001) over a median follow-up of 19 months. The median RTB total score improved from 9 to 4 (P < .0001). There was no impact of RTB total score (<3 vs >3) on VCSS (P = NS), GOS (P = NS), VAS pain score (P = NS) or CIVIQ-20 score (P = NS) at the various time points. Concurrent stenting was used in 23 limbs (33%) and delayed stenting was carried out in 10 limbs (14%). The median time to delayed stenting was 4 months after the initial thrombus removal intervention. CONCLUSIONS: In patients undergoing PMT or PMT with catheter-directed thrombolysis for acute symptomatic IFDVT, the restoration of inline flow seems to be adequate to provide symptom relief and decrease the incidence of PTS. The extent of RTB does not seem to impact the VCSS, GOS, VAS pain score, or quality of life after such restoration. Stenting can be pursued selectively in the acute setting to help restore inline flow.

Comparison of intravascular ultrasound and magnetic resonance venography in the diagnosis of chronic iliac venous disease.

BACKGROUND: The diagnosis of chronic iliofemoral venous obstruction (CIVO) can be made with several different modalities. Intravascular ultrasound (IVUS) examination is the gold standard in the diagnosis of CIVO. However, being invasive, it should not be the initial examination to screen patients with CIVO. The aim of this report is to compare the performance of magnetic resonance venography (MRV) with IVUS examination in the diagnosis of CIVO. METHODS: From January 2016 to December 2020, the records of all patients who underwent preoperative MRV and then IVUS in the evaluation of CIVO were analyzed retrospectively. RESULTS: There were 505 patients who were evaluated by any modality for CIVO. Of these patients, 15% (78) were evaluated by MRV. Patients who had failed a trial of conservative therapy for at least 3 to 6 months and who had disabling and lifestyle-limiting symptoms of CIVO were selected to undergo further evaluation with MRV at the treating physician's discretion. For inclusion in analysis, technically satisfactory IVUS examination and MRV data were mandatory. Data was available for 60 common iliac vein (CIV) segments and 61 external iliac vein (EIV) segments for comparative analysis after appropriate exclusions. The mean age of the patients was 56 ± 15 years. The male to female ratio was 1:2. The distribution of patients across different CEAP classes was as follows: CEAP 3, 28%; CEAP 4, 62%; CEAP 5, 2%; and CEAP 6, 8%. Bland-Altman plots of the mean difference in area between IVUS examination and MRI were 74.1% for CIV and 56.9% for EIV. The sensitivity of MRV was 93% and 100%, and the specificity was 0% and 50% for CIV and EIV, respectively. The positive predictive value was 93% and 86%; the negative predictive value was 0 and 50% for CIV and EIV, respectively. Improvement was noted in clinical parameters (Venous Clinical Severity Score, visual analog pain scale, and grade of swelling) after IVUS examination and stenting after MRV. For the Venous Clinical Severity Score, the score improved from 6.0 ± 2.7 (before the procedure) to 4 ± 2.7 (after the procedure) (P = .0001). CONCLUSIONS: There is dimensional disparity between MRV and IVUS examination in the diagnosis of symptomatic CIVO. MRV has a high sensitivity but low specificity when compared with IVUS examination and overestimates the severity of the stenosis in both the EIV and CIV. MRV is not a reliable diagnostic tool for iliac vein stenosis and should not be used for the definitive disposition of patients with CIVO.

On the recovery of disorders of consciousness under intrathecal baclofen administration for severe spasticity-An observational study.

BACKGROUND: Occasionally, patients show dramatic recovery from disorders of consciousness (DOC) under intrathecal baclofen (ITB), an established treatment option for severe supraspinal spasticity. Anecdotal explanations for ITB-related recovery of cognition include modulation of afferent impulses at the spinal level, thereby reducing spasticity-related proprioceptive information overload within cortico-thalamo-cortical connections. OBJECTIVE: In this retrospective patient chart analysis, we assessed whether a reduction in spasticity would be associated with an increase in Coma Recovery Scale revised (CRS-R) scores in a larger sample of patients than previously published. METHODS: From a hospital-based ITB treatment register, we extracted data from 26 patients with DOC and severe supraspinal spasticity who improved by >2 points on the Coma Recovery Scale revised (CRS-R) within 6 months after ITB treatment initiation. We assessed Modified Ashworth scale (MAS) scores and CRS-R scores on admission (PRE) and 3 and 6 months after initiation of ITB treatment (3M, 6M). We performed correlation analysis of the scores and their respective changes (PRE to 3M, 3M to 6M). We also correlated the time from acute event until ITB initiation to CRS-R scores at 3M and 6M. RESULTS: ITB led to significant improvement in spasticity based on MAS scores, which did not correlate to the improvements seen in CRS-R total and subscale scores. Daily ITB dose did neither correlate to MAS scores nor to CRS-total scores in the whole patient group, but after 3 months, ITB dose correlated to some CRS-R subscale scores in some patient subgroups. Time until ITB treatment did not correlate to CRS-R scores later on. CONCLUSIONS: Our data confirm that ITB may exert beneficial effects in selected DOC patients with respect to improved cognitive functions, which, however, do not correlate to its antispastic effect. The lack of correlation between time to ITB and CRS-R outcome, but significant CRS-R improvements following pump implantation, renders spontaneous remissions unlikely and leaves room for alternative pharmacological mechanisms.

Cross-Seeding Controls Aß Fibril Populations and Resulting Functions.

Amyloid peptides nucleate from monomers to aggregate into fibrils through primary nucleation. Pre-existing fibrils can then act as seeds for additional monomers to fibrillize through secondary nucleation. Both nucleation processes occur simultaneously, yielding a distribution of fibril polymorphs that can generate a spectrum of neurodegenerative effects. Understanding the mechanisms driving polymorph structural distribution during both nucleation processes is important for uncovering fibril structure-function relationships, as well as for creating polymorph distributions in vitro that better match fibril structures found in vivo. Here, we explore how cross-seeding wild-type (WT) Aß1-40 with Aß1-40 mutants E22G (Arctic) and E22Δ (Osaka), as well as with WT Aß1-42, affects the distribution of fibril structural polymorphs and how changes in structural distribution impact toxicity. Transmission electron microscopy analysis revealed that fibril seeds derived from mutants of Aß1-40 imparted their structure to WT Aß1-40 monomers during secondary nucleation, but WT Aß1-40 fibril seeds do not affect the structure of fibrils assembled from mutant Aß1-40 monomers, despite the kinetic data indicating accelerated aggregation when cross-seeding of any combination of mutants. Additionally, WT Aß1-40 fibrils seeded with mutant fibrils produced similar structural distributions to the mutant seeds with similar cytotoxicity profiles. This indicates that mutant fibril seeds not only impart their structure to growing WT Aß1-40 aggregates but also impart cytotoxic properties. Our findings establish a relationship between the fibril structure and the phenotype on a polymorph population level and that these properties can be passed on through secondary nucleation to the succeeding generations of fibrils.

Long-term improvement of limb reflux prevalence and severity after iliac vein stent placement.

BACKGROUND: The effect of iliac vein stenting on ipsilateral limb reflux is unknown and has remained a matter of speculation. It has been suggested that the propensity for reflux might worsen when proximal stenosis is corrected. This could allow for retrograde flow with coughing and the Valsalva maneuver, stressing the valve. We examined this hypothesis by an analysis of the long-term effects of iliac vein stenting on limb reflux using a single-center, retrospective analysis of prospectively collected data. METHODS: Reflux data from duplex ultrasound of 1387 limbs in 1228 patients who had undergone iliac vein stenting from 1997 to 2018 were analyzed. Of the 1387 limbs, 632 (46%) had had ipsilateral duplex ultrasound-determined valve reflux before stenting, and 747 limbs (54%) had not had reflux; data were missing for 8 limbs. Reflux status before and after stenting was available for seven individual segments for each limb in the database for analysis (total, 9653 segments). The stented patients were examined for reflux at least annually during the follow-up period (range, 1-26 years). Segmental reflux prevalence was detected using duplex ultrasound. We have referred to this as "duplex reflux" or simply "reflux." Reflux severity was graded using (1) a reflux segmental score, assigning one point each for refluxing segments in the limb; (2) air plethysmography (venous filling index [VFI90]); and (3) ambulatory venous pressure (venous filling time [VFT]). RESULTS: Prestent duplex reflux was present in a combination of superficial, deep, and perforator segments. Reflux prevalence ranged from 7% of deep femoral segments to 51% at the popliteal segment. Post-stent reflux resolution varied from 21% at the femoral vein segment to 58% at the perforator segments. Reflux had completely resolved in 23% of the limbs. New-onset reflux was rare, with a median incidence of 7% for all segments at risk, with cumulative improvement (Kaplan-Meier curve) in reflux severity (segment score, VFI90, and VFT) for most limbs. These metrics were unimproved, with residual reflux in only 18%, 11%, and 6% (segment score, VFI90, and VFT, respectively) of the limbs at long-term follow-up. CONCLUSIONS: Long-term follow-up of limbs after iliac vein stenting has shown that the associated ipsilateral reflux prevalence and severity will improve in most limbs over time.

Ultraviolet Photodissociation Dynamics of the Cyclohexyl Radical: The H-Atom Product Channel.

The ultraviolet (UV) photodissociation dynamics of the jet-cooled cyclohexyl (c-C6H11) radical is studied using the high-n Rydberg atom time-of-flight (HRTOF) technique. The cyclohexyl radical is produced by the 193 nm photodissociation of chlorocyclohexane and bromocyclohexane and is examined in the photolysis wavelength region of 232-262 nm. The H-atom photofragment yield (PFY) spectrum contains a broad peak centered at 250 nm, which is in good agreement with the UV absorption spectrum of the cyclohexyl radical and assigned to the 3p Rydberg states. The translational energy distributions of the H-atom loss product channel, P(ET)'s, are bimodal, with a slow (low ET) component peaking at ∼6 to 7 kcal/mol and a fast (high ET) component peaking at ∼44-48 kcal/mol. The fraction of the average translational energy in the total excess energy, ⟨fT⟩, is in the range of 0.16-0.25 in the photolysis wavelength region of 232-262 nm. The H-atom product angular distribution of the slow component is isotropic, while that of the fast component is anisotropic with an anisotropy parameter of ß ≈ 0.5-0.7. The bimodal product translational energy and angular distributions indicate two dissociation pathways to the H + C6H10 products in cyclohexyl. The high-ET anisotropic component is from a repulsive, prompt dissociation on a repulsive potential energy surface coupling with the Rydberg excited states to produce H + cyclohexene. The low-ET isotropic component is consistent with the unimolecular dissociation of hot radical on the ground electronic state after internal conversion from the Rydberg states. The similarity of the photodissociation dynamics of the cyclohexyl radical to the previously studied small linear and branched alkyls expands on the understanding of the dissociation dynamics of alkyl radicals to include larger cyclic alkyl radicals.

Distal organ inflammation and injury after resuscitative endovascular balloon occlusion of the aorta in a porcine model of severe hemorrhagic shock.

BACKGROUND AND OBJECTIVE: Resuscitative Endovascular Balloon Occlusion of Aorta (REBOA) has emerged as a potential life-saving maneuver for the management of non-compressible torso hemorrhage in trauma patients. Complete REBOA (cREBOA) is inherently associated with the burden of ischemia reperfusion injury (IRI) and organ dysfunction. However, the distal organ inflammation and its association with organ injury have been little investigated. This study was conducted to assess these adverse effects of cREBOA following massive hemorrhage in swine. METHODS: Spontaneously breathing and consciously sedated Sinclair pigs were subjected to exponential hemorrhage of 65% total blood volume over 60 minutes. Animals were randomized into 3 groups (n = 7): (1) Positive control (PC) received immediate transfusion of shed blood after hemorrhage, (2) 30min-cREBOA (A30) received Zone 1 cREBOA for 30 minutes, and (3) 60min-cREBOA (A60) given Zone 1 cREBOA for 60 minutes. The A30 and A60 groups were followed by resuscitation with shed blood post-cREBOA and observed for 4h. Metabolic and hemodynamic effects, coagulation parameters, inflammatory and end organ consequences were monitored and assessed. RESULTS: Compared with 30min-cREBOA, 60min-cREBOA resulted in (1) increased IL-6, TNF-α, and IL-1ß in distal organs (kidney, jejunum, and liver) (p < 0.05) and decreased reduced glutathione in kidney and liver (p < 0.05), (2) leukopenia, neutropenia, and coagulopathy (p < 0.05), (3) blood pressure decline (p < 0.05), (4) metabolic acidosis and hyperkalemia (p < 0.05), and (5) histological injury of kidney and jejunum (p < 0.05) as well as higher levels of creatinine, AST, and ALT (p < 0.05). CONCLUSION: 30min-cREBOA seems to be a feasible and effective adjunct in supporting central perfusion during severe hemorrhage. However, prolonged cREBOA (60min) adverse effects such as distal organ inflammation and injury must be taken into serious consideration.

Ultraviolet photodissociation dynamics of the n-butyl, s-butyl, and t-butyl radicals.

Photodissociation dynamics of the jet-cooled n-butyl radical via the 3s Rydberg state and the s-butyl radical via the 3p Rydberg states in the ultraviolet region of 233 nm-258 nm, as well as the t-butyl radical via the 3d Rydberg states at 226 nm-244 nm, are studied using the high-n Rydberg atom time-of-flight technique. The H-atom photofragment yield spectra of the n-butyl, s-butyl, and t-butyl radicals show a broad feature centered around 247 nm, 244 nm, and 234 nm, respectively. The translational energy distributions of the H + C4H8 products, P(ET)'s, of the three radicals are bimodal, with a slow (low ET) component peaking at ∼6 kcal/mol and a fast (high ET) component peaking at ∼52 kcal/mol-57 kcal/mol, ∼43 kcal/mol, and ∼37 kcal/mol for n-butyl, s-butyl, and t-butyl, respectively. The fraction of the products' translational energy in the available energy, ⟨ fT⟩, is 0.31, 0.30, and 0.27 for n-butyl, s-butyl, and t-butyl, respectively. The H-atom product angular distributions of the slow component are isotropic for all three radicals, while those of the fast component are anisotropic for n-butyl and s-butyl with an anisotropy parameter ß âˆ¼ 0.7 and ∼ 0.3 and that of the fast component of t-butyl is nearly isotropic. The bimodal product translational energy and angular distributions indicate two dissociation pathways to the H + C4H8 products in these three radicals, a direct, prompt dissociation on the repulsive potential energy surface coupling with the Rydberg excited states, and a unimolecular dissociation of the hot radical on the ground electronic state after internal conversion from the Rydberg states.

Functionalized Mesoporous Silicas Direct Structural Polymorphism of Amyloid-ß Fibrils.

The aggregation of amyloid-ß (Aß) is associated with the onset of Alzheimer's disease (AD) and involves a complex kinetic pathway as monomers self-assemble into fibrils. A central feature of amyloid fibrils is the existence of multiple structural polymorphs, which complicates the development of disease-relevant structure-function relationships. Developing these relationships requires new methods to control fibril structure. In this work, we evaluated the effect that mesoporous silicas (SBA-15) functionalized with hydrophobic (SBA-PFDTS) and hydrophilic groups (SBA-PEG) have on the aggregation kinetics and resulting structure of Aß1-40 fibrils. The hydrophilic SBA-PEG had little effect on amyloid kinetics, while as-synthesized and hydrophobic SBA-PFDTS accelerated aggregation kinetics. Subsequently, we quantified the relative population of fibril structures formed in the presence of each material using electron microscopy. Fibrils formed from Aß1-40 exposed to SBA-PEG were structurally similar to control fibrils. In contrast, Aß1-40 incubated with SBA-15 or SBA-PFDTS formed fibrils with shorter crossover distances that were more structurally representative of fibrils found in AD patient derived samples. Overall, our results suggest that mesoporous silicas and other exogenous materials are promising scaffolds for the de novo production of specific fibril polymorphs of Aß1-40 and other amyloidogenic proteins.