Cardiac radiation improves ventricular function in mice and humans with cardiomyopathy.

BACKGROUND: Rapidly dividing cells are more sensitive to radiation therapy (RT) than quiescent cells. In the failing myocardium, macrophages and fibroblasts mediate collateral tissue injury, leading to progressive myocardial remodeling, fibrosis, and pump failure. Because these cells divide more rapidly than cardiomyocytes, we hypothesized that macrophages and fibroblasts would be more susceptible to lower doses of radiation and that cardiac radiation could therefore attenuate myocardial remodeling. METHODS: In three independent murine heart failure models, including models of metabolic stress, ischemia, and pressure overload, mice underwent 5 Gy cardiac radiation or sham treatment followed by echocardiography. Immunofluorescence, flow cytometry, and non-invasive PET imaging were employed to evaluate cardiac macrophages and fibroblasts. Serial cardiac magnetic resonance imaging (cMRI) from patients with cardiomyopathy treated with 25 Gy cardiac RT for ventricular tachycardia (VT) was evaluated to determine changes in cardiac function. FINDINGS: In murine heart failure models, cardiac radiation significantly increased LV ejection fraction and reduced end-diastolic volume vs. sham. Radiation resulted in reduced mRNA abundance of B-type natriuretic peptide and fibrotic genes, and histological assessment of the LV showed reduced fibrosis. PET and flow cytometry demonstrated reductions in pro-inflammatory macrophages, and immunofluorescence demonstrated reduced proliferation of macrophages and fibroblasts with RT. In patients who were treated with RT for VT, cMRI demonstrated decreases in LV end-diastolic volume and improvements in LV ejection fraction early after treatment. CONCLUSIONS: These results suggest that 5 Gy cardiac radiation attenuates cardiac remodeling in mice and humans with heart failure. FUNDING: NIH, ASTRO, AHA, Longer Life Foundation.

Evaluating the Use of Graph Neural Networks and Transfer Learning for Oral Bioavailability Prediction.

Oral bioavailability is a pharmacokinetic property that plays an important role in drug discovery. Recently developed computational models involve the use of molecular descriptors, fingerprints, and conventional machine-learning models. However, determining the type of molecular descriptors requires domain expert knowledge and time for feature selection. With the emergence of the graph neural network (GNN), models can be trained to automatically extract features that they deem important. In this article, we exploited the automatic feature selection of GNN to predict oral bioavailability. To enhance the prediction performance of GNN, we utilized transfer learning by pre-training a model to predict solubility and obtained a final average accuracy of 0.797, an F1 score of 0.840, and an AUC-ROC of 0.867, which outperformed previous studies on predicting oral bioavailability with the same test data set.

Human Cardiac Pericytes Are Susceptible to SARS-CoV-2 Infection.

COVID-19 is associated with serious cardiovascular complications, with incompletely understood mechanism(s). Pericytes have key functions in supporting endothelial cells and maintaining vascular integrity. We demonstrate that human cardiac pericytes are permissive to SARS-CoV-2 infection in organotypic slice and primary cell cultures. Viral entry into pericytes is mediated by endosomal proteases, and infection leads to up-regulation of inflammatory markers, vasoactive mediators, and nuclear factor kappa-B-dependent cell death. Furthermore, we present evidence of cardiac pericyte infection in COVID-19 myocarditis patients. These data demonstrate that human cardiac pericytes are susceptible to SARS-CoV-2 infection and suggest a role for pericyte infection in COVID-19.

Perceptions associated with the public attitudes toward epilepsy (PATE) scale: A mixed-method study.

BACKGROUND: In epilepsy stigma, certain perceptions are culturally dependent and greatly influence a person's attitudes. Hence, we aimed to explore the perceptions associated with attitudes toward epilepsy in various urban subpopulations. METHOD: This is a mixed-method study employing the Public Attitude Toward Epilepsy (PATE) scale as the quantitative measure, followed by a semi-structured interview. The qualitative data were then counted and analyzed concurrently with the quantitative data. RESULT: A total of 410 respondents (104 people with epilepsy [PWE]; 104 family members [FM]; 100 medical students [MS]; 102 public [Pb]) aged 37 years (IQR 23-55) were recruited. They were mostly female (57.3%), Chinese (52.0%), and highly educated (63.7%). The attitudes toward epilepsy among medical students are the best, followed by the PWE and their family members, and the worst among the public. The qualitative results revealed 4 main themes, which were "general social values", "epilepsy severity and control", "PWE's abilities", and "harms and burdens to the respondents and others". A two-dimensional perception model was constructed based on these themes, which consisted of general-personal and universal-specific dimensions. Generally, the PWE/FM subgroup focused more on PWE's abilities, whereas the MS/Pb subgroup more on general social values, and harms and burden. In the education aspect, most attitudes were related to the epilepsy severity and PWE's abilities, whereas in employment, the main consideration was the PWE's abilities. Burden to life and concern about inheritance were major considerations in the marital relationship. Those with positive attitudes tend to highlight the importance of general social values, while negative attitudes associated more with epilepsy severity. In general domain, general social values were the main considering factor but in personal domain, most participants will consider epilepsy severity and control, harms and burden to themselves. CONCLUSION: The perceptions underlying attitudes toward epilepsy were complex and varied between subpopulations, attitude levels, domains, and aspects of life. (304 words).

An analysis of preoperative shoulder and neck balance and surgical outcome in 111 adolescent idiopathic scoliosis patients with two subtypes of Lenke 1 curves.

OBJECTIVE: The incidence of postoperative shoulder imbalance following posterior spinal fusion (PSF) is still high in Lenke 1 curves despite following current treatment recommendations for upper instrumented vertebra (UIV) selection. The objective of this retrospective study was to identify differences in preoperative shoulder balance and to report the surgical outcome of two subtypes of Lenke 1 curves (flexible vs stiff) in patients with adolescent idiopathic scoliosis (AIS). METHODS: The authors grouped patients' curves as Lenke 1-ve (flexible) when their preoperative proximal thoracic side bending (PTSB) Cobb angle was < 15° and as Lenke 1+ve (stiff) when the PTSB Cobb angle was 15°-24.9°. The authors hypothesized that these two subtypes had distinct preoperative and postoperative shoulder and neck balance following PSF using pedicle screw constructs. RESULTS: Fifty patients had Lenke 1 (flexible) curves and 61 had Lenke 1 (stiff) curves. The mean preoperative T1 tilt for patients with Lenke 1 (flexible) was -4.9° ± 5.3°, and for those with Lenke 1 (stiff) curves it was -1.0° ± 5.3° (p < 0.001). Mean cervical axis (CA) was -0.1° ± 3.2° for Lenke 1 (flexible) curves and 2.3° ± 3.5° for Lenke 1 (stiff) curves (p < 0.001). Preoperative radiographic shoulder height (RSH) and clavicle angle (Cla-A) were similar between the two curve subtypes. Following surgery, there were significant differences between the subtypes in terms of T1 tilt (p < 0.001), RSH (p = 0.014), and Cla-A (p = 0.031). Interestingly, 41.0% of patients with a Lenke 1 (stiff) curve had +ve T1 tilt compared to 2.0% in Lenke 1 (flexible) group. Moreover, 26.2% of patients with the Lenke 1 (stiff) curve had +ve RSH compared to 12.0% of those with Lenke 1 (flexible) curves. And, 24.6% of patients with Lenke 1 (stiff) had +ve Cla-A compared to 10.0% of those with Lenke 1 (flexible) curves. CONCLUSIONS: Lenke 1 (flexible) and Lenke 1 (stiff) curves had distinct preoperative T1 tilt and CA measurements. Following PSF, the authors noted +ve T1 tilt in 41% of patients with Lenke 1 (stiff) curves versus 2.0% in those with Lenke 1 (flexible) curves. The authors also noted a significant difference in postoperative RSH and Cla-A measurements.

Conformity and Changes in the Radiological Neck and Shoulder Balance Parameters Throughout 3-Year Follow-up Period: Do They Remain the Same?

STUDY DESIGN: Retrospective study. OBJECTIVE: The primary objective of this study was to assess the conformity of the radiological neck and shoulder balance parameters throughout a follow-up period of more than 2 years. SUMMARY OF BACKGROUND DATA: Postoperative shoulder and neck imbalance are undesirable features among Adolescent Idiopathic Scoliosis patients who underwent Posterior Spinal Fusion. There are many clinical and radiological parameters used to assess this clinical outcome. However, we do not know whether these radiological parameters conform throughout the entire follow-up period. METHODS: This was a retrospective study done in a single academic institution. Inclusion criteria were patients with scoliosis who underwent posterior instrumented spinal fusion with pedicle screw fixation and attended all scheduled follow-ups for at least 24 months postoperatively. Radiological shoulder parameters were measured from both preoperative antero-posterior and postoperative antero-posterior radiographs. Lateral shoulder parameters were: Radiographic Shoulder Height, Clavicle Angle (Cla-A), Clavicle-Rib Intersection Difference, and Coracoid Height Difference. Medial shoulder and neck parameters were: T1 Tilt and Cervical Axis (CA). RESULTS: The radiographs of 50 patients who had surgery done from November 2013 to November 2015 were analyzed. Mean age of this cohort was 16.3 ±â€Š7.0 years. There were 38 (76%) female patients and 12 (24%) male patients. Mean final follow-up was 38.6 ±â€Š5.8 months. When conformity assessment of the radiological parameter using the interclass coefficient correlation was done, we found that all parameters had significant correlation (P < 0.05). T1 tilt (0.78) had good reliability, CA (0.47), Clavicle-Rib Intersection Difference (0.43), and Coracoid Height Difference (0.40) had moderate reliability and Radiographic Shoulder Height (0.22) and Cla-A (0.13) had poor reliability. CONCLUSION: All the radiological shoulder and neck balance parameters studied were conformed and suitable to be used to assess the patient postoperatively. Amongst these radiological parameters, T1 tilt followed by CA recorded to be the most reliable parameters over time. LEVEL OF EVIDENCE: 4.

The Reliability of Intraoperative Crossbar Technique in Determining the Upper Instrumented Vertebra (UIV) Tilt Angle for Adolescent Idiopathic Scoliosis (AIS) Undergoing Posterior Spinal Fusion.

STUDY DESIGN: Retrospective study. OBJECTIVE: This study looked into whether crossbar can reliably measure Upper Instrumend Vertebra (UIV) tilt angle intraoperatively and accurately predict the UIV tilt angle postoperatively and at final follow-up. SUMMARY OF BACKGROUND DATA: Postoperative shoulder imbalance is a common cause of poor cosmetic appearance leading to patient dissatisfaction. There were no reports describing the technique or method in measuring the UIV tilt angle intraoperatively. Therefore, this study was designed to look into the reliability and accuracy of the usage of intraoperative crossbar in measuring the UIV tilt angle intraoperatively. METHODS: Lenke 1 and 2 Adolescent Idiopathic Scoliosis patients who underwent instrumented Posterior Spinal Fusion using pedicle screw constructs with minimum follow-up of 24 months were recruited for this study. After surgical correction, intraoperative UIV tilt angle was measured using a crossbar. Immediate postoperative and final follow up UIV tilt angle was measured on the standing anteroposterior radiographs. RESULTS: A total of 100 patients were included into this study. The reliability of the intraoperative crossbar to measure the optimal UIV tilt angle intraoperatively was determined by repeated measurements by assessors and measurement by different assessors. We found that the intra observer and inter observer reliability was very good with intraclass correlation coefficient values of >0.9. The accuracy of the intraoperative crossbar to measure the optimal UIV tilt angle intraoperatively was determined by comparing this measurement with the postoperative UIV tilt angle. We found that there was no significant difference (P>0.05) between intraoperative, immediate postoperative, and follow-up UIV tilt angle. CONCLUSIONS: The crossbar can be used to measure the intraoperative UIV tilt angle consistently and was able to predict the postoperative UIV tilt angle. It was a cheap, simple, reliable, and accurate instrument to measure the intraoperative UIV tilt angle.

How Common Is Medial and Lateral Shoulder Discordance in Lenke 1 and 2 Curves?: A Preoperative Analysis of Medial and Lateral Shoulder Balance Among 151 Lenke 1 and 2 Adolescent Idiopathic Scoliosis Patients.

STUDY DESIGN: Retrospective study of prospectively collected data. OBJECTIVE: To analyze the incidence and the patterns of medial and lateral shoulder discordance among Lenke 1 and 2 patients. SUMMARY OF BACKGROUND DATA: Postoperative shoulder imbalance (PSI) is still common in Lenke 1 and 2 adolescent idiopathic scoliosis (AIS). This could be due to presence of medial dan lateral shoulder discordance. METHODS: One hundred fifty-one Lenke 1 and 2 AIS patients were recruited. Lenke 1 curves were subclassified into Lenke 1-ve (flexible) (proximal thoracic side bending [PTSB] Cobb angle <15°) and 1+ve curves (stiff) (PTSB Cobb angle between 15° and 24.9°). T1 tilt represented "medial shoulder balance" and radiological shoulder height (RSH) represented "lateral shoulder balance." We categorized patients into three concordant shoulder pattern types (medial balanced/lateral balanced [MBLB], medial imbalanced+ve/lateral imbalanced+ve (MI+ve/LI+ve), medial imbalanced-ve/lateral Imbalanced-ve (MI-ve/LI-ve), and six discordant shoulder pattern types. RESULTS: The mean age was 16.2 ±â€Š5.7 years. Eighty-one patients (53.6%) had concordant pattern and 70 patients (46.4%) had discordant pattern. Lateral shoulder imbalance was noted in 35.1% of patients and medial shoulder imbalance in 43.7% of patients. In Lenke 1-ve curves, 35 patients (68.6%) had concordant shoulder imbalance with medial imbalanced-ve/lateral imbalanced-ve (MI-ve/LI-ve) being the commonest pattern (68.6%). In Lenke 1+ve curves, 33 patients (55.0%) had concordant shoulder pattern with medial balanced/lateral balanced (MB/LB) being the commonest type (57.6%). In Lenke 2 AIS, 27 patients (67.5%) had discordant pattern with medial imbalanced+ve/lateral balanced (MI+ve/LB) being the commonest pattern (44.4%) (P value = 0.002). CONCLUSION: 46.4% Lenke 1 and 2 AIS patients had shoulder discordant pattern. This was more prevalent in Lenke 2 curves (67.5%). In Lenke 1-ve (flexible) curves, MI-ve/LI-ve pattern was the commonest pattern. In Lenke 1+ve curves (stiff), there were almost equal number of concordant and discordant shoulder pattern. In Lenke 2 patients, the most common pattern was MI+ve/LB. LEVEL OF EVIDENCE: 4.

Radiological and clinical outcome of selective thoracic fusion for patients with Lenke 1C and 2C adolescent idiopathic scoliosis with a minimum follow-up of 2 years.

BACKGROUND CONTEXT: In Lenke 1C and 2C curves, the choice between selective thoracic fusion (STF) versus non-selective thoracic fusion as the optimal surgical treatment is controversial. OBJECTIVE: This study aimed to assess the radiological and clinical outcome of patients with Lenke 1C and 2C curves treated with STF. STUDY DESIGN: This is a retrospective study. PATIENT SAMPLE: A total of 44 patients comprised the study sample. METHODS: Forty-four patients with Lenke 1C and 2C curves with adolescent idiopathic scoliosis who underwent STF were reviewed. Radiological parameters and Scoliosis Research Society (SRS)-22r scores were assessed preoperatively, postoperatively, and on final follow-up. The incidence of coronal decompensation, lumbar decompensation, and adding-on phenomenon were reported. RESULTS: Mean follow-up duration was 45.1±12.3 months and mean age was 17.0±5.1 years. The preoperative middle thoracic and thoracolumbar/lumbar (MT:TL/L) Cobb angle ratio was 1.4±0.3 and the MT:TL/L apical vertebra translation (AVT) ratio was 1.6±0.8. Final follow-up coronal balance was -13.0±11.5 mm, main thoracic AVT was 6.9±11.8 mm, and lumbar AVT was -20.4±13.8 mm (p<.05). Lumbar Cobb angle improved from 47.5°±7.8° to 24.9°±8.2° after operation and 23.3°±9.8° at final follow-up. The spontaneous lumbar curve correction rate was 50.9%. There were 9 patients (20.5%) who had coronal decompensation, 4 patients (9.1%) who had lumbar decompensation, and 11 patients (25.0%) who had adding-on phenomenon. We did not perform any revision surgery. The SRS-22r scores improved significantly in the overall scores, self-image, and mental health domain. CONCLUSIONS: Selective thoracic fusion led to improvement in the radiological and clinical outcome for patients with Lenke 1C and 2C. Although no patients required revision surgery, the rate of coronal decompensation, lumbar decompensation, and adding-on phenomenon are significant.

How type II CRISPR-Cas establish immunity through Cas1-Cas2-mediated spacer integration.

CRISPR (clustered regularly interspaced short palindromic repeats) and the nearby Cas (CRISPR-associated) operon establish an RNA-based adaptive immunity system in prokaryotes. Molecular memory is created when a short foreign DNA-derived prespacer is integrated into the CRISPR array as a new spacer. Whereas the RNA-guided CRISPR interference mechanism varies widely among CRISPR-Cas systems, the spacer integration mechanism is essentially identical. The conserved Cas1 and Cas2 proteins form an integrase complex consisting of two distal Cas1 dimers bridged by a Cas2 dimer. The prespacer is bound by Cas1-Cas2 as a dual-forked DNA, and the terminal 3'-OH of each 3' overhang serves as an attacking nucleophile during integration. The prespacer is preferentially integrated into the leader-proximal region of the CRISPR array, guided by the leader sequence and a pair of inverted repeats inside the CRISPR repeat. Spacer integration in the well-studied Escherichia coli type I-E CRISPR system also relies on the bacterial integration host factor. In type II-A CRISPR, however, Cas1-Cas2 alone integrates spacers efficiently in vitro; other Cas proteins (such as Cas9 and Csn2) have accessory roles in the biogenesis phase of prespacers. Here we present four structural snapshots from the type II-A system of Enterococcus faecalis Cas1 and Cas2 during spacer integration. Enterococcus faecalis Cas1-Cas2 selectively binds to a splayed 30-base-pair prespacer bearing 4-nucleotide 3' overhangs. Three molecular events take place upon encountering a target: first, the Cas1-Cas2-prespacer complex searches for half-sites stochastically, then it preferentially interacts with the leader-side CRISPR repeat, and finally, it catalyses a nucleophilic attack that connects one strand of the leader-proximal repeat to the prespacer 3' overhang. Recognition of the spacer half-site requires DNA bending and leads to full integration. We derive a mechanistic framework to explain the stepwise spacer integration process and the leader-proximal preference.

Structure Basis for Directional R-loop Formation and Substrate Handover Mechanisms in Type I CRISPR-Cas System.

Type I CRISPR systems feature a sequential dsDNA target searching and degradation process, by crRNA-displaying Cascade and nuclease-helicase fusion enzyme Cas3, respectively. Here we present two cryo-EM snapshots of the Thermobifida fusca type I-E Cascade: (1) unwinding 11 bp of dsDNA at the seed-sequence region to scout for sequence complementarity, and (2) further unwinding of the entire protospacer to form a full R-loop. These structures provide the much-needed temporal and spatial resolution to resolve key mechanistic steps leading to Cas3 recruitment. In the early steps, PAM recognition causes severe DNA bending, leading to spontaneous DNA unwinding to form a seed-bubble. The full R-loop formation triggers conformational changes in Cascade, licensing Cas3 to bind. The same process also generates a bulge in the non-target DNA strand, enabling its handover to Cas3 for cleavage. The combination of both negative and positive checkpoints ensures stringent yet efficient target degradation in type I CRISPR-Cas systems.