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OBJECTIVES:. Anemia has been associated with an increased risk of both cardiac arrest and stroke, frequent complications of COVID-19. The effect of hemoglobin level at ICU admission on a composite outcome of cardiac arrest or stroke in an international cohort of COVID-19 patients was investigated. DESIGN:. Retrospective analysis of prospectively collected database. SETTING:. A registry of COVID-19 patients admitted to ICUs at over 370 international sites was reviewed for patients diagnosed with cardiac arrest or stroke up to 30 days after ICU admission. Anemia was defined as: normal (hemoglobin ≥ 12.0 g/dL for women, ≥ 13.5 g/dL for men), mild (hemoglobin 10.0–11.9 g/dL for women, 10.0–13.4 g/dL for men), moderate (hemoglobin ≥ 8.0 and < 10.0 g/dL for women and men), and severe (hemoglobin < 8.0 g/dL for women and men). PATIENTS:. Patients older than 18 years with acute COVID-19 infection in the ICU. INTERVENTIONS:. None. MEASUREMENTS AND MAIN RESULTS:. Of 6926 patients (median age = 59 yr, male = 65%), 760 patients (11.0%) experienced stroke (2.0%) and/or cardiac arrest (9.4%). Cardiac arrest or stroke was more common in patients with low hemoglobin, occurring in 12.8% of patients with normal hemoglobin, 13.3% of patients with mild anemia, and 16.7% of patients with moderate/severe anemia. Time to stroke or cardiac arrest by anemia status was analyzed using Cox proportional hazards regression with death as a competing risk. Covariates selected through clinical knowledge were age, sex, comorbidities (diabetes, hypertension, obesity, and cardiac or neurologic conditions), pandemic era, country income, mechanical ventilation, and extracorporeal membrane oxygenation. Moderate/severe anemia was associated with a higher risk of cardiac arrest or stroke (hazard ratio, 1.32; 95% CI, 1.05–1.67). CONCLUSIONS:. In an international registry of ICU patients with COVID-19, moderate/severe anemia was associated with increased hazard of cardiac arrest or stroke.

The COVID-19 pandemic and its accompanying restrictions have significantly impacted people’s lives globally. There is an increasing interest in examining the influence of this unprecedented situation on our mental well-being, with less attention towards the impact of the elongation of COVID-19-related measures on youth with a pre-existing psychiatric/developmental disorder. The majority of studies focus on individuals, such as students, adults, and youths, among others, with little attention being given to the elongation of COVID-19-related measures and their impact on a special group of individuals, such as children and adolescents with diagnosed developmental and psychiatric disorders. In addition, most of these studies adopt statistical methodologies to identify pair-wise relationships among factors, an approach that limits the ability to understand and interpret the impact of various factors. In response, this study aims to adopt an explainable machine learning approach to identify factors that explain the deterioration or amelioration of mood state in a youth clinical sample. The purpose of this study is to identify and interpret the impact of the greatest contributing features of mood state changes on the prediction output via an explainable machine learning pipeline. Among all the machine learning classifiers, the Random Forest model achieved the highest effectiveness, with 76% best AUC-ROC Score and 13 features. The explainability analysis showed that stress or positive changes derived from the imposing restrictions and COVID-19 pandemic are the top two factors that could affect mood state.

Abstract Global Climate Model tuning (calibration) is a tedious and time‐consuming process, with high‐dimensional input and output fields. Experts typically tune by iteratively running climate simulations with hand‐picked values of tuning parameters. Many, in both the statistical and climate literature, have proposed alternative calibration methods, but most are impractical or difficult to implement. We present a practical, robust, and rigorous calibration approach on the atmosphere‐only model of the Department of Energy's Energy Exascale Earth System Model (E3SM) version 2. Our approach can be summarized into two main parts: (a) the training of a surrogate that predicts E3SM output in a fraction of the time compared to running E3SM, and (b) gradient‐based parameter optimization. To train the surrogate, we generate a set of designed ensemble runs that span our input parameter space and use polynomial chaos expansions on a reduced output space to fit the E3SM output. We use this surrogate in an optimization scheme to identify values of the input parameters for which our model best matches gridded spatial fields of climate observations. To validate our choice of parameters, we run E3SMv2 with the optimal parameter values and compare prediction results to expertly‐tuned simulations across 45 different output fields. This flexible, robust, and automated approach is straightforward to implement, and we demonstrate that the resulting model output matches present day climate observations as well or better than the corresponding output from expert tuned parameter values, while considering high‐dimensional output and operating in a fraction of the time.

Abstract Background Knee osteoarthritis (KOA) is a common degenerative joint condition that causes disability and pain in the elderly population. The prevalence of KOA among persons aged 63 or above is approximately 30%. Previous studies have reported the positive effects of Tui-na treatment and the Chinese herbal formula Du-Huo-Ji-Sheng Decoction (DHJSD) for KOA treatment. The current study aims to evaluate the add-on therapeutic effect of oral administration of DHJSD on KOA in addition to Tui-na. Methods We conducted a prospective, randomized, controlled clinical trial. Seventy study subjects with KOA were randomly assigned to the treatment and control groups in a 1:1 ratio. Both two groups received eight sessions of Tui-na manipulation for 4 weeks. The DHJSD was only administered to the study subjects in the treatment group. The primary outcome measure was rated using the WOMAC at the end of treatment (4 weeks). Secondary outcomes were assessed using EQ-5D-5L, a health-related quality of life with 5-level EQ-5D version at end of treatment (week 4) and follow-up (week 8). Results No statistically significant difference was found between two groups on WOMAC scores at the end of treatment. The mean WOMAC Pain subscale score was significantly lower in the treatment group than control group at week 8 follow up (mean difference, MD − 1.8, 95% CI − 3.5 to − 0.02, P = 0.048). The mean WOMAC Stiffness subscale score was significantly lower in the treatment group than in the control group at week 2 (MD 0.74, 95% CI 0.05 to 1.42, P = 0.035) and week 8 follow up (MD 0.95, 95% CI 0.26 to 1.65, P = 0.008). The mean EQ-5D index value was significantly improved in the treatment group than in the control group at week 2 (MD 0.17, 95% CI 0.02 to 0.31, P = 0.022). The analysis of WOMAC scores and EQ-5D-5L in both groups showed statistically significant improvement with time. No significant adverse effect was found during the trial. Conclusion DHJSD may have an add-on effect in addition to Tui-na manipulation relieving pain and improving stiffness as well as quality of life (QOL) in patients with KOA. The combined treatment was generally safe and well tolerated. Trial registration The study was registered at the ClinicalTrials.gov (website: https://clinicaltrials.gov/ct2/show/NCT04492670 , registry number: NCT04492670), registered on 30 July 2020.

The COVID-19 pandemic has affected how clinical examinations are conducted, resulting in the Royal College of Psychiatrists delivering the Clinical Assessment of Skills and Competence virtually. Although this pragmatic step has allowed for progression of training, it has come at the cost of a significantly altered examination experience. This article aims to explore the fairness of such an examination, the difference in trainee experience, and the use of telemedicine to consider what might be lost as well as gained at a time when medical education and delivery of healthcare are moving toward the digitised frontier.

Abstract Background The Basic Local Alignment Search Tool (BLAST) is a suite of commonly used algorithms for identifying matches between biological sequences. The user supplies a database file and query file of sequences for BLAST to find identical sequences between the two. The typical millions of database and query sequences make BLAST computationally challenging but also well suited for parallelization on high-performance computing clusters. The efficacy of parallelization depends on the data partitioning, where the optimal data partitioning relies on an accurate performance model. In previous studies, a BLAST job was sped up by 27 times by partitioning the database and query among thousands of processor nodes. However, the optimality of the partitioning method was not studied. Unlike BLAST performance models proposed in the literature that usually have problem size and hardware configuration as the only variables, the execution time of a BLAST job is a function of database size, query size, and hardware capability. In this work, the nucleotide BLAST application BLASTN was profiled using three methods: shell-level profiling with the Unix “time” command, code-level profiling with the built-in “profiler” module, and system-level profiling with the Unix “gprof” program. The runtimes were measured for six node types, using six different database files and 15 query files, on a heterogeneous HPC cluster with 500+ nodes. The empirical measurement data were fitted with quadratic functions to develop performance models that were used to guide the data parallelization for BLASTN jobs. Results Profiling results showed that BLASTN contains more than 34,500 different functions, but a single function, RunMTBySplitDB, takes 99.12% of the total runtime. Among its 53 child functions, five core functions were identified to make up 92.12% of the overall BLASTN runtime. Based on the performance models, static load balancing algorithms can be applied to the BLASTN input data to minimize the runtime of the longest job on an HPC cluster. Four test cases being run on homogeneous and heterogeneous clusters were tested. Experiment results showed that the runtime can be reduced by 81% on a homogeneous cluster and by 20% on a heterogeneous cluster by re-distributing the workload. Discussion Optimal data partitioning can improve BLASTN’s overall runtime 5.4-fold in comparison with dividing the database and query into the same number of fragments. The proposed methodology can be used in the other applications in the BLAST+ suite or any other application as long as source code is available.

Purpose – This study aims to explain the effect of entrepreneurial orientation (EO) dimensions on firms’ export performance. The study has considered three dimensions of EO: innovativeness, proactiveness and risk-taking. Export performance has been measured through multifaceted determinants: financial, strategic and satisfaction levels of exporting firms. Design/methodology/approach – To explain the association between such determinants, qualitative face-to-face, in-depth interviews were conducted with the respondents from the apparel manufacturing companies of Bangladesh. The study has also applied a content analysis technique to explore and triangulate the descriptions of EO dimensions on performance. Findings – The research results reveal that EO has a meaningful explanation of export performance from the lived experience of the respondents. The study has also found frequent explanations and indicators about the association of innovativeness and proactiveness on export performance, but the risk-taking dimension has revealed no effects. Research limitations/implications – The research has different implications on theoretical, managerial and policy-making aspects of EO and export performance. Nevertheless, the findings are based on the opinions of a small sample of entrepreneurs and high-level managers. Thus, it is important to test the generalizability of these findings in future research using a larger sample. Originality/value – This study indicated product, process and market creation innovativeness. Risks related to product innovation, new market entry and politics are considered for risk-taking measures, while proactive innovation, technology and new market searching are used for measuring proactiveness. Additionally, multifaceted performance determinants have been applied to evaluate export performance. Hence, this study is a unique qualitative analysis with important theoretical, practical and methodological implications.

In recent years, cyberbullying on social media sites has increased among adolescents and even adults. While there are many factors and forms of cyberbullying, schoolchildren are vulnerable groups that are exposed to cyberbullying threats to their mental and emotional health. On the other hand, teachers have the responsibility in the place of the parent while at school, and they need to manage the issue at bay. This study focuses on the identification and analysis of teachers’ advice about cyberbullying, thus leading to the conceptualization of a decision-making model for the school to guide other teachers on this matter. This study is an explorative case study conducted in a prominent, semi-private international school in Lingyi, Shandong Province, China. With the purposive sampling approach, thirty-two teachers volunteered in this study; to recap and describe their encounters through writing in an online survey form. As analysis, their responses were validated for trustworthiness, while codes are checked through member checking, and eventually thematically analyzed to address the research questions. Findings from this study revealed a list of tactics that were categorized into three themes: (a) Preventive measures; (b) Counter-measures, and (c) Corrective measures. Each of these categories highlights specific ways that teachers suggest their students to when confronted with cyberbullying. Thereafter, each category was integrated as a model of decision-making in cyberbullying for improvements to the standard operating protocol of the researched school. In addition, this study proposes more similar research in other school settings so that more advisory tactics can be accumulated to enable education policymakers, school administrators, and teachers to make better decisions to manage issues associated with cyberbullying.

Background: A preclinical animal model is an imperative tool for uncovering and understanding the tumourigenic hallmarks of human ovarian cancer; the disease is often lethal because it is commonly diagnosed in the advanced stage, where widespread cancer nodules mainly reside within peritoneal regions. Mouse models as a xenograft tumour host or genetic manipulation ovarian cancer-derived mice are widely used for studying specific hypothesis rationale in ovarian cancer. However, limited information associated with disease progression is obtained from such studies; whether it is the best model to study advanced ovarian cancer phenotype or suitable preclinical biomarkers for detecting and monitoring ovarian cancer progression is under study. This study used an ID-8 syngeneic mouse ovarian cancer model with immunocompetence. We monitored cancer growth and development using combination modalities of cancer-specific cancer antigen-125 (CA-125), interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF) blood markers, which are well-known for their association with tumour progression in humans. Methods: Ten C57/BL6 female mice were intraperitoneally implanted with ID-8 Trp53 wild-type and monitored the progression of the tumour, until mice developed clinical ascites. Blood was taken at the time of intraperitoneal (IP) implantation (Day 0) and then collected weekly, and levels of biomarkers were analysed with enzyme-linked immunosorbent assay (ELISA). In addition, tumour tissue was collected and proceeded with histological staining. Results: We found that blood biomarkers CA-125, IL-6 and VEGF were not readily correlated with tumour progression. However, these biomarkers were markedly elevated in ascitic fluid at the advanced stage of the disease. Conclusions: We conclude that blood biomarkers in a syngeneic mouse model are, to some extent, not readily found in the blood as opposed to human ovarian cancer. Model anatomical and physiological differences between rodents and humans might explain this discrepancy.

Posttraumatic osteoarthritis (PTOA) arises secondary to joint injuries and is characteristically driven by inflammatory mediators. PTOA is often studied in the setting of ACL tears. However, a wide range of other injuries also lead to PTOA pathogenesis. The purpose of this study was to characterize the morphological changes in the uninjured ACL in a PTOA inflammatory environment. We retrospectively reviewed 14 ACLs from 13 Yucatan minipigs, 7 of which had undergone our modified intra-articular drilling (mIAD) procedure, which induced PTOA through inflammatory mediators. Seven ACLs were harvested from mIAD minipigs (PTOA) and seven ACLs from control minipigs with no cartilage degeneration (non-PTOA). ACL degeneration was evaluated using histological scoring systems. IL-1β, NF-κB, and TNF-α mRNA expression in the synovium was measured using qRT-PCR. PTOA minipigs demonstrated significant ACL degeneration, marked by a disorganized extracellular matrix, increased vascularity, and changes in cellular shape, density, and alignment. Furthermore, IL-1β, NF-κB, and TNF-α expression was elevated in the synovium of PTOA minipigs. These findings demonstrate the potential for ACL degeneration in a PTOA environment and emphasize the need for anti-inflammatory disease-modifying therapies following joint injury.

The concept of pedagogical leadership (PL) is vague and still developing in the field of Educational Leadership. This article examines the body of knowledge on this elusive term, to extract the key components that constitute the studies of PL in the last five years (2017-2020). The keyword of pedagogical leader* was solely used to search in the Web of Science database, and subsequently narrowed down to 54 reviewed articles through listed criteria for bibliometric analysis. Apart from the global bibliometric descriptions of PL, it is evident that the term pedagogical leadership is an emerging theme in WoS databases and is mostly cited from Nordic and European countries. Future suggestions for eastern scholars are to collaborate with western scholars, to better understand its concepts as compared to other prominent types of leadership such as Transformational, Distributed, and Instructional Leadership.

(1) Purpose: The objective was to evaluate CT perfusion and radiomic features for prediction of one year disease free survival in laryngeal and hypopharyngeal cancer. (2) Method and Materials: This retrospective study included pre and post therapy CT neck studies in 36 patients with laryngeal/hypopharyngeal cancer. Tumor contouring was performed semi-autonomously by the computer and manually by two radiologists. Twenty-six radiomic features including morphological and gray-level features were extracted by an internally developed and validated computer-aided image analysis system. The five perfusion features analyzed included permeability surface area product (PS), blood flow (flow), blood volume (BV), mean transit time (MTT), and time-to-maximum (Tmax). One year persistent/recurrent disease data were obtained following the final treatment of definitive chemoradiation or after total laryngectomy. We performed a two-loop leave-one-out feature selection and linear discriminant analysis classifier with generation of receiver operating characteristic (ROC) curves and confidence intervals (CI). (3) Results: 10 patients (28%) had recurrence/persistent disease at 1 year. For prediction, the change in blood flow demonstrated a training AUC of 0.68 (CI 0.47–0.85) and testing AUC of 0.66 (CI 0.47–0.85). The best features selected were a combination of perfusion and radiomic features including blood flow and computer-estimated percent volume changes-training AUC of 0.68 (CI 0.5–0.85) and testing AUC of 0.69 (CI 0.5–0.85). The laryngoscopic percent change in volume was a poor predictor with a testing AUC of 0.4 (CI 0.16–0.57). (4) Conclusions: A combination of CT perfusion and radiomic features are potential predictors of one-year disease free survival in laryngeal and hypopharyngeal cancer patients.

Despite the importance of neural-network quantum states, representing fermionic matter is yet to be fully achieved. Here the authors map fermionic degrees of freedom to spin ones and use neural-networks to perform electronic structure calculations on model diatomic molecules to achieve chemical accuracy.

Feed-forward neural networks are a novel class of variational wave functions for correlated many-body quantum systems. Here, we propose a specific neural network ansatz suitable for systems with real-valued wave functions. Its characteristic is to encode the all-important rugged sign structure of a quantum wave function in a convolutional neural network with discrete output. Its training is achieved through an evolutionary algorithm. We test our variational ansatz and training strategy on two spin-1/2 Heisenberg models, one on the two-dimensional square lattice and one on the three-dimensional pyrochlore lattice. In the former, our ansatz converges with high accuracy to the analytically known sign structures of ordered phases. In the latter, where such sign structures are a priori unknown, we obtain better variational energies than with other neural network states. Our results demonstrate the utility of discrete neural networks to solve quantum many-body problems.

This article focuses on a Participatory Action Research (PAR) project that was carried out in a Malaysian Public University during three series of lockdowns during the Covid-19 pandemic. During that time, all higher education institutes in Malaysia were barred from conducting face-to-face lectures that resorted to fully online learning throughout two semesters during the academic session of 2020 to 2021. As an educator, I was primary concerned about their achievement in learning outcomes and knowledge transfer through online learning of the Entrepreneurship Education (EE) Course. Therefore, this PAR described my critical reflections on my students’ feedbacks from the analysis of my online video lessons, online student forums, and open-ended online surveys regarding their sentiments and perceptions on their learning experience. Ultimately, this PAR is to help me improve my teaching skills, to highlight gaps in the course content and suggest strategies of delivery in the future. As such, this article concludes by highlighting the mismatch between learning gaps, the extent the EE course learning outcomes address the students’ personal obstacles in entrepreneurship and my future strategies to mitigate the challenges of entrepreneurship education.

Achievement of fit between the residual limb and prosthetic socket during socket manufacture is a priority for clinicians and is essential for safety. Clinicians have recognised the potential benefits of having a sensor system that can provide objective socket-limb interface pressure measurements during socket fitting, but the cost of existing systems makes current technology prohibitive. This study will report on the characterisation, validation and preliminary clinical implementation of a low cost, portable, wireless sensor system designed for use during socket manufacture. Characterisation and benchtop testing demonstrated acceptable accuracy, behaviour at variable temperature, and dynamic response for use in prosthetic socket applications. Our sensor system was validated with simultaneous measurement by a commercial sensor system in the sockets of three transtibial prosthesis users during a fitting session in the clinic. There were no statistically significant differences between the sensor system and the commercial sensor for a variety of functional activities. The sensor system was found to be valid in this clinical context. Future work should explore how pressure data relates to ratings of fit and comfort, and how objective pressure data might be used to assist in clinical decision making.

Sarcopenia is a state of catabolic muscle wasting prevalent in geriatric patients. Likewise, osteoarthritis is an age-related musculoskeletal disease affecting patients with similar demographics. Late-stage hip osteoarthritis is often treated with total hip arthroplasty (THA). As sarcopenia influences the surgical outcomes, this study aimed to assess the impact of sarcopenia on the outcomes of THA. A 1:3 matched case–control study of sarcopenic to control patients was performed using a large national database. In total, 3992 patients were analyzed. Sarcopenic patients undergoing THA were more likely to experience dislocation (odds ratio (OR) = 2.19, 95% confidence interval (CI) 1.21–3.91) within 1 year of THA. Furthermore, sarcopenic patients had higher urinary tract infection rates (OR = 1.79, CI 1.32–2.42) and a greater risk of 90-day hospital readmission (hazard ratio (HR) = 1.39, CI 1.10–1.77). Sarcopenic patients experienced more falls (OR = 1.62, CI 1.10–2.39) and fragility fractures (OR = 1.77, CI 1.34–2.31). Similarly, sarcopenic patients had higher day of surgery costs (USD 13,534 vs. USD 10,504) and 90-day costs (USD 17,139 vs. USD 13,394) compared with the controls. Ultimately, sarcopenic patients undergoing THA experience higher rates of postoperative complications and incur greater medical costs. Given the potential risks, orthopedic surgeons may consider treating or reducing the severity of sarcopenia before surgery.

IMPORTANCE:. Factors associated with mortality in coronavirus disease 2019 patients on invasive mechanical ventilation are still not fully elucidated. OBJECTIVES:. To identify patient-level parameters, readily available at the bedside, associated with the risk of in-hospital mortality within 28 days from commencement of invasive mechanical ventilation or coronavirus disease 2019. DESIGN, SETTING, AND PARTICIPANTS:. Prospective observational cohort study by the global Coronavirus Disease 2019 Critical Care Consortium. Patients with laboratory-confirmed coronavirus disease 2019 requiring invasive mechanical ventilation from February 2, 2020, to May 15, 2021. MAIN OUTCOMES AND MEASURES:. Patient characteristics and clinical data were assessed upon ICU admission, the commencement of invasive mechanical ventilation and for 28 days thereafter. We primarily aimed to identify time-independent and time-dependent risk factors for 28-day invasive mechanical ventilation mortality. RESULTS:. One-thousand five-hundred eighty-seven patients were included in the survival analysis; 588 patients died in hospital within 28 days of commencing invasive mechanical ventilation (37%). Cox-regression analysis identified associations between the hazard of 28-day invasive mechanical ventilation mortality with age (hazard ratio, 1.26 per 10-yr increase in age; 95% CI, 1.16–1.37; p < 0.001), positive end-expiratory pressure upon commencement of invasive mechanical ventilation (hazard ratio, 0.81 per 5 cm H2O increase; 95% CI, 0.67–0.97; p = 0.02). Time-dependent parameters associated with 28-day invasive mechanical ventilation mortality were serum creatinine (hazard ratio, 1.28 per doubling; 95% CI, 1.15–1.41; p < 0.001), lactate (hazard ratio, 1.22 per doubling; 95% CI, 1.11–1.34; p < 0.001), Paco2 (hazard ratio, 1.63 per doubling; 95% CI, 1.19–2.25; p < 0.001), pH (hazard ratio, 0.89 per 0.1 increase; 95% CI, 0.8–14; p = 0.041), Pao2/Fio2 (hazard ratio, 0.58 per doubling; 95% CI, 0.52–0.66; p < 0.001), and mean arterial pressure (hazard ratio, 0.92 per 10 mm Hg increase; 95% CI, 0.88–0.97; p = 0.003). CONCLUSIONS AND RELEVANCE:. This international study suggests that in patients with coronavirus disease 2019 on invasive mechanical ventilation, older age and clinically relevant variables monitored at baseline or sequentially during the course of invasive mechanical ventilation are associated with 28-day invasive mechanical ventilation mortality hazard. Further investigation is warranted to validate any causative roles these parameters might play in influencing clinical outcomes.

Abstract Aim To validate the 4‐ and 7‐point Chinese Non‐pharmacological Therapy Experience Scales and test the psychometric properties of the scales on persons with intellectual disability. Design A validation study. Methods Sixty‐seven persons with intellectual disability were recruited from six hostels or centres for persons with intellectual disability in Hong Kong. A total of 1,163 and 1,161 observations were collected by the trained observers with the 4‐point and 7‐point scales, respectively. The floor and ceiling effects, inter‐rater reliability, internal consistency, responsiveness of both scales and the scale equivalence were examined. Results The Cronbach's α of the 4‐ and 7‐point scales was .762 and .797, respectively. The correlation between the two scales was 0.906. The inter‐rater reliability of the 4‐ and 7‐point scales was 0.774 and 0.835, respectively. Neither scale had the floor or ceiling effects. The effect size of the 7‐point scale was consistently higher than that of the 4‐point scale.

ABSTRACTMolecular surveillance of infections is essential in monitoring their transmission in the population. In this study, newly diagnosed HIV patients' phylogenetic, clinical and behavioural data were integrated, and an information diffusion model was incorporated in analysing transmission dynamics. A genetic network was constructed from HIV sequences, from which transmission cascades were extracted. From the transmission cascades, CRF01_AE had higher values of information diffusion metrics, including scale, speed and range, than that of B, signifying the distinct transmission patterns of two circulating subtypes in Hong Kong. Patients connected in the network, were more likely male, younger, of main circulating subtypes, to have acquired HIV infection locally, and a higher CD4 level at diagnosis. Genetic connections varied among men who have sex with men (MSM) who used different channels of sex networking and varied in their engagement in risk behaviours. MSM using recreational drugs for sex held positions of greater importance within the network. Significant differences in network metrics were observed among MSM as differentiated by their mobile apps usage patterns, evidencing the impact of social network on transmission networks. The applied model in the presence of consistently collected longitudinal data could enhance HIV molecular epidemiologic surveillance for informing future intervention planning.

We study the dissipative dynamics of a periodically driven inhomogeneous critical lattice model in one dimension. The closed system dynamics starting from pure initial states is well-described by a driven Conformal Field Theory (CFT), which predicts the existence of both heating and non-heating phases in such systems. Heating is inhomogeneous and is manifested via the emergence of black-hole like horizons in the system. The robustness of this CFT phenomenology when considering thermal initial states and open systems remains elusive. First, we present analytical results for the Floquet CFT time evolution for thermal initial states. Moreover, using exact calculations of the time evolution of the lattice density matrix, we demonstrate that for short and intermediate times, the closed system phase diagram comprising heating and non-heating phases, persists for thermal initial states on the lattice. Secondly, in the fully open system with boundary dissipators, we show that the nontrivial spatial structure of the heating phase survives particle-conserving and non-conserving dissipations through clear signatures in mutual information and energy density evolution.

Introduction Coronavirus 2019 (SARS-CoV-2) infection in children occurred in Peru as of March 2020, leading to pediatric patients’ hospitalization in areas adapted for this purpose at the Edgardo Rebagliati Martins National Hospital. In the beginning, the demand for hospitalization was low, but it increased gradually. Consistent with international reports, the majority of patients presented mild or moderate symptoms. Nonetheless, there were also severe cases, even fatal ones. Objectives To describe the characteristics and clinical outcome of pediatric patients with COVID-19 hospitalized in a referral hospital in Lima, Peru, between March and August 2020. Methods A descriptive and inferential cross-sectional study was carried out. The population includes all hospitalized patients in the Department of Pediatrics, with clinical and surgical diagnoses associated with COVID-19. Results We included 100 patients, with an average age of 83.4 ± 54 months, with a predominance of male patients (55%). Hospitalized patients were grouped into five categories: respiratory failure (17%), multisystemic inflammatory syndrome (MIS-C) (31%), neurological presentation (19%), acute abdomen (20%), and patients with oncological problems (13%). Most of the patients (74%) had comorbidities. Regarding the presenting symptoms, intestinal pain predominated in the appendicitis group (90%, p < 0.001), fever was present in most patients with respiratory failure (64.7%); multisystemic inflammatory syndrome (90.3%), neurological manifestations (15.8%), acute abdomen (50%) and oncological conditions (61.5%) were also present in these patients. Kawasaki symptoms were found in 38.7% of the patients with multisystemic inflammatory syndrome. Mortality was 4%. Respiratory problems (29.4%) and multisystemic inflammatory syndrome (22.6%) required admission to intensive care, more frequently than the other presentations (p = 0.008). Conclusions We conclude that the vulnerability in the pediatric population is the one that has preexisting conditions. We divided our patients according to presentation, diagnosis, and complications, which were predominantly respiratory. We also had oncological patients with COVID-19.

Background: Macrophage activation syndrome (MAS) is characterized by excessive activation of macrophages and lymphocytes, leading to multiorgan dysfunction. As the initial manifestation of systemic lupus erythematosus (SLE), MAS is rare in children. Due to the COVID-19 pandemic, it is vital to identify the MAS as it shares similar characteristics with the multisystem inflammatory syndrome in children (MIS-C). Case report: We report the case of an 11-year-old male adolescent with symptoms of MIS-C. Although with negative results of RT-PCR (reverse transcription-polymerase chain reaction) and serology for SARS-CoV-2, contact with a positive COVID-19 relative was reported. When admitted to a referral hospital center, the patient received standard treatment for MIS-C. Although the same scheme was given on three occasions, the patient showed no response to initial therapy. Thus, the patient was classified as a refractory case. When the study was extended to other differential diagnoses, we found MAS associated with SLE. Therefore, the patient was treated with etoposide, cyclosporine, dexamethasone, and methotrexate and showed a good clinical response. Conclusions: MAS associated with SLE is rare in the pediatric population. MAS shares inflammatory markers with the MIS-C and is often confused with rheumatologic, infectious, and neoplastic entities. Reporting this case is important to identify differential diagnoses in patients presenting as MIS-C and decide on timely treatment, as it could be harmful or even fatal if a definitive diagnosis is not obtained on time.

BackgroundSelf-focused attention and safety behaviours are both associated with adolescent social anxiety. In adults, experimental studies have indicated that the processes are causally implicated in social anxiety, but this hypothesis has not yet been tested in a youth sample.MethodsThis experiment explored this possibility by asking high and low socially anxious adolescents (N = 57) to undertake conversations under different conditions. During one conversation they were instructed to focus on themselves and use safety behaviours, and in the other they focused externally and did not use safety behaviours. Self-report, conversation partner report and independent assessor ratings were taken.ResultsSelf-focus and safety behaviours increased feelings and appearance of anxiety and undermined performance for all participants, but only high socially anxious participants reported habitually using self-focus and safety behaviours.ConclusionsThe findings provide support for the causal role of self-focus and safety behaviours in adolescent social anxiety and point to the potential clinical value of techniques reversing them to treat the disorder.

Social anxiety is associated with reduced educational achievement. Given that concentration is a predictor of educational achievement, and social anxiety symptoms are associated with reduced concentration in class, this prospective study examined the possibility that social anxiety may impair educational achievement through reduced classroom concentration. A sample of 509 participants (53.8% female; M age: 12.77 years [SD = 0.81]) recruited from secondary schools completed questionnaires assessing social anxiety symptoms, depressive symptoms, and concentration in class. Educational achievement was assessed by internal grades within schools. An indirect effect of social anxiety on later educational achievement via concentration was observed, over and above baseline achievement and depression symptoms; adolescents with higher levels of social anxiety tend to have more difficulties concentrating in class, which in turn is associated with poorer academic outcomes. Findings underscore the challenges socially anxious adolescents will face trying to learn in school, and the need for education providers and clinicians to consider the effect of social anxiety symptoms on concentration and learning.

Reliable posture labels in hospital environments can augment research studies on neural correlates to natural behaviors and clinical applications that monitor patient activity. However, many existing pose estimation frameworks are not calibrated for these unpredictable settings. In this paper, we propose a semi-automated approach for improving upper-body pose estimation in noisy clinical environments, whereby we adapt and build around an existing joint tracking framework to improve its robustness to environmental uncertainties. The proposed framework uses subject-specific convolutional neural network models trained on a subset of a patient's RGB video recording chosen to maximize the feature variance of each joint. Furthermore, by compensating for scene lighting changes and by refining the predicted joint trajectories through a Kalman filter with fitted noise parameters, the extended system yields more consistent and accurate posture annotations when compared with the two state-of-the-art generalized pose tracking algorithms for three hospital patients recorded in two research clinics.

The spin-1/2 Heisenberg model on the pyrochlore lattice is an iconic frustrated three-dimensional spin system with a rich phase diagram. Besides hosting several ordered phases, the model is debated to possess a spin-liquid ground state when only nearest-neighbor antiferromagnetic interactions are present. Here, we contest this hypothesis with an extensive numerical investigation using both exact diagonalization and complementary variational techniques. Specifically, we employ a resonating-valence-bond-like, many-variable, Monte Carlo ansatz and convolutional neural network quantum states for (variational) calculations with up to 4×4^{3} and 4×3^{3} spins, respectively. We demonstrate that these techniques yield consistent results, allowing for reliable extrapolations to the thermodynamic limit. We consider the (λ,j_{2}/j_{1}) parameter space, with j_{2}, j_{1} being nearest and next-to-nearest neighbor interactions and λ the XXZ interaction anisotropy. Our main results are (1) the determination of the phase transition between the putative spin-liquid phase and the neighboring magnetically ordered phase and (2) a careful characterization of the ground state in terms of symmetry-breaking tendencies. We find clear indications of a dimer order with spontaneously broken inversion and rotational symmetry, calling the scenario of a featureless quantum spin liquid into question. Our work showcases how many-variable variational techniques can be used to make progress in answering challenging questions about three-dimensional frustrated quantum magnets.

BackgroundThe identification of negative self-imagery as a maintenance factor in adult social anxiety has led to enhanced treatments for this population. Whilst intrusive negative imagery is commonly reported by socially anxious adolescents, no studies have demonstrated that it plays a causal role in maintaining symptoms. To assess this possibility, we undertook an experimental study manipulating social self-imagery in high socially anxious adolescents.MethodsHigh socially anxious adolescents undertook two conversations under different conditions. During one conversation they held a negative social self-image in mind, and in the other they held a benign social self-image in mind. Self-report, conversation partner report and independent assessor ratings were taken.ResultsWhen participants held a negative self-image in mind, they reported feeling more anxious, and believed they looked more anxious and performed more poorly. Furthermore, they overestimated how anxious they looked compared to conversation partner ratings. As well as distorting participants' perception of their anxious appearance, holding a negative image in mind also had observable effects on the interaction. Participants were rated as looking more anxious and performing less well by their conversation partner when they held such images in mind, and the conversation was rated more critically by conversation partners and independent observers. Finally, a preliminary mediation analysis suggested that the detrimental effect of negative imagery on the social interaction may be partly due to the spontaneous use of avoidant safety behaviours.ConclusionsThe findings provide support for a causal role of negative self-imagery in adolescent social anxiety and point to the potential clinical value of techniques targeting imagery to treat the disorder.

We introduce NetKet, a comprehensive open source framework for the study of many-body quantum systems using machine learning techniques. The framework is built around a general and flexible implementation of neural-network quantum states, which are used as a variational ansatz for quantum wavefunctions. NetKet provides algorithms for several key tasks in quantum many-body physics and quantum technology, namely quantum state tomography, supervised learning from wavefunction data, and ground state searches for a wide range of customizable lattice models. Our aim is to provide a common platform for open research and to stimulate the collaborative development of computational methods at the interface of machine learning and many-body physics. Keywords: Neural-network quantum states, Variational Monte Carlo, Quantum state tomography, Machine learning, Supervised learning

We study the heating dynamics of a generic one-dimensional critical system when driven quasiperiodically. Specifically, we consider a Fibonacci drive sequence comprising the Hamiltonian of uniform conformal field theory (CFT) describing such critical systems and its sine-square deformed counterpart. The asymptotic dynamics is dictated by the Lyapunov exponent which has a fractal structure embedding Cantor lines where the exponent is exactly zero. Away from these Cantor lines, the system typically heats up fast to infinite energy in a nonergodic manner where the quasiparticle excitations congregate at a small number of select spatial locations resulting in a buildup of energy at these points. Periodic dynamics with no heating for physically relevant timescales is seen in the high-frequency regime. As we traverse the fractal region and approach the Cantor lines, the heating slows enormously and the quasiparticles completely delocalize at stroboscopic times. Our setup allows us to tune between fast and ultraslow heating regimes in integrable systems.

While driven interacting quantum matter is generically subject to heating and scrambling, certain classes of systems evade this paradigm. We study such an exceptional class in periodically driven critical (1+1)-dimensional systems with a spatially modulated but disorder-free time evolution operator. Instead of complete scrambling, the excitations of the system remain well defined. Their propagation is analogous to the evolution along light cones in a curved spacetime obtained by two black holes. The Hawking temperature serves as an order parameter which distinguishes between heating and nonheating phases. Beyond a timescale determined by the inverse Hawking temperature, excitations are absorbed by the black holes resulting in a singular concentration of energy at their horizon. We obtain these results analytically within conformal field theory, capitalizing on a mapping to sine-squared deformed field theories. Furthermore, by means of numerical calculations for an interacting XXZ spin-1/2 chain, we demonstrate that our findings survive lattice regularization.

Background: advanced ovarian cancer often presents with ascites. These ascites contain small clusters of cancer cells, which may contribute greatly to the metastatic potential of ovarian cancer in the peritoneal cavity. Therefore, understanding the unique protein expressions of this cell population will provide vital information for the development of tailored, targeted treatment. In this study, we isolate floating ovarian cancer cells from ovarian cancer patient ascitic fluid and use these cells to document that the expression of EGFR/HER-2 proteins may be essential for the growth and survival of these floating cancer cell clusters. Methods: ascitic fluid-derived cells were isolated from ascitic fluid by using Ficoll separation. Cells were cultured in a non-adherent condition for six days. The protein level of EGFR, HER-2, AKT, and ERK and their phosphorylation in ovarian cancer cell lines were determined by immunofluorescence. The immunofluorescent staining for proteins presented in ascitic fluid-derived cells determined the intensity profile of each protein using Carl Zeiss Blue software. Results: Isolated ovarian cancer cells from ascitic fluid have a measurable level of EGFR and HER-2 proteins. The inhibition of EGFR and EGFR/HER-2 positive cells with gefitinib and canertinib selectively disrupts cell viability and the protein level of EGFR, HER-2, AKT and ERK and their respective phosphorylation status. In addition, the dual EGFR/HER-2 inhibitor canertinib demonstrates greater anti-tumour effects than gefitinib in EGFR/HER-2 positive cells. Conclusion: These studies reveal an important role of multiple activation of receptor tyrosine kinases in floating ovarian cancer cells, as well as the importance of a dual EGFR/HER-2 inhibitor used as alternative adjuvant therapy in advanced ovarian cancer patients.

The threat of viral pandemics demands a comprehensive understanding of evolution at the host-pathogen interface. Here, we show that the accessibility of adaptive mutations in influenza nucleoprotein at fever-like temperatures is mediated by host chaperones. Particularly noteworthy, we observe that the Pro283 nucleoprotein variant, which (1) is conserved across human influenza strains, (2) confers resistance to the Myxovirus resistance protein A (MxA) restriction factor, and (3) critically contributed to adaptation to humans in the 1918 pandemic influenza strain, is rendered unfit by heat shock factor 1 inhibition-mediated host chaperone depletion at febrile temperatures. This fitness loss is due to biophysical defects that chaperones are unavailable to address when heat shock factor 1 is inhibited. Thus, influenza subverts host chaperones to uncouple the biophysically deleterious consequences of viral protein variants from the benefits of immune escape. In summary, host proteostasis plays a central role in shaping influenza adaptation, with implications for the evolution of other viruses, for viral host switching, and for antiviral drug development.

BackgroundWhile HIV incidence among men who have sex with men (MSM) is increasing in Hong Kong, unprotected sex apparently remains prevalent among those infected but virally non-suppressed. Little is known about how sexual behaviours and sexual connections may change among MSM along their HIV care continuum.MethodsIn this retrospective cross-sectional study, HIV-positive MSM attending the largest HIV specialist clinic in Hong Kong between October and December 2014 were invited to complete a self-administrated structured questionnaire. Their behavioural profile and partner sourcing patterns during the one-year period respectively (a) before HIV diagnosis, (b) after HIV diagnosis, (c) after initiation of antiretroviral treatment and (d) preceding the survey were examined.ResultsOf 345 recruited MSM, 304 (88.1%) had treatment initiated and 272 (78.8%) had viral load suppressed. In the first year after HIV diagnosis, the proportion reporting inconsistent condom use dropped from 47.0% to 17.5% (pConclusionsMost HIV-positive MSM had persistently low level of sexual risk behaviours along their care continuum and achieved viral load suppression, conferring a general reduction of secondary transmission risk in Hong Kong. To increase the effectiveness of Treatment as Prevention strategy, uptake of HIV testing for undiagnosed HIV-positive MSM shall be emphasised.

We propose an exact construction for atypical excited states of a class of nonintegrable quantum many-body Hamiltonians in one, two, and three dimensions that display area-law entanglement entropy. These examples of many-body “scar” states have, by design, other properties, such as topological degeneracies, usually associated with the gapped ground states of symmetry-protected topological phases or topologically ordered phases of matter.

We study the heat and spin transport properties in a ring of interacting spins coupled to heat baths at different temperatures. We show that interactions, by inducing avoided crossings, can be a means to tune both the total heat current flowing between the ring and the baths, and the way it flows through the system. In particular, we recognize three regimes in which the heat current flows clockwise, counterclockwise, and in parallel. The temperature bias between the baths also induces a spin current within the ring, whose direction and magnitude can be tuned by the interaction. Lastly, we show how the ergotropy of the nonequilibrium steady state can increase significantly near the avoided crossings.

Resveratrol has aroused significant scientific interest as it has been claimed that it exhibits a spectrum of health benefits. These include effects as an anti-inflammatory and an antitumour compound. The purpose of this study was to investigate and compare any potential antigrowth effects of resveratrol and two of its derivatives, acetyl-resveratrol and polydatin, on 3D cell aggregates of the EGFR/Her-2 positive and negative ovarian cancer cell lines SKOV-3 and OVCAR-8, respectively. Results showed that resveratrol and acetyl-resveratrol reduced cell growth in the SKOV-3 and OVCAR-8 in a dose-dependant manner. The growth reduction was mediated by the induction of apoptosis via the cleavage of poly(ADP-ribose) polymerase (PARP-1). At lower concentrations, 5 and 10 µM, resveratrol, acetyl-resveratrol, and polydatin were less effective than higher concentrations, 50 and 100 µM. In SKOV-3 line, at higher concentrations, resveratrol and polydatin significantly reduced the phosphorylation of Her-2 and EGFR and the expression of Erk. Acetyl-resveratrol, on the other hand, did not change the activation of Her-2 and EGFR. Resveratrol, acetyl-resveratrol, and polydatin suppressed the secretion of VEGF in a dose-dependant fashion. In the OVCAR-8 cell line, resveratrol and acetyl-resveratrol at 5 and 10 µM increased the activation of Erk. Above these concentrations they decreased activation. Polydatin did not produce this effect. This study demonstrates that resveratrol and its derivatives may inhibit growth of 3D cell aggregates of ovarian cancer cell lines via different signalling molecules. Resveratrol and its derivatives, therefore, warrant further in vivo evaluation to assess their potential clinical utility.

The men-having-sex-with-men (MSM) population has become one of the major risk groups for HIV-1 infection in the Asia Pacific countries. Hong Kong is located in the centre of Asia and the transmission history of HIV-1 subtype B transmission among MSM remained unclear. The aim of this study was to investigate the transmission dynamics of HIV-1 subtype B virus in the Hong Kong MSM population. Samples of 125 HIV-1 subtype B infected MSM patients were recruited in this study. Through this study, the subtype B epidemic in the Hong Kong MSM population was identified spreading mainly among local Chinese who caught infection locally. On the other hand, HIV-1 subtype B infected Caucasian MSM caught infection mainly outside Hong Kong. The Bayesian phylogenetic analysis also indicated that 3 separate subtype B epidemics with divergence dates in the 1990s had occurred. The first and latest epidemics were comparatively small-scaled; spreading among the local Chinese MSM while sauna-visiting was found to be the major sex partner sourcing reservoir for the first subtype B epidemic. However, the second epidemic was spread in a large-scale among local Chinese MSM with a number of them having sourced their sex partners through the internet. The epidemic virus was estimated to have a divergence date in 1987 and the infected population in Hong Kong had a logistic growth throughout the past 20 years. Our study elucidated the evolutionary and demographic history of HIV-1 subtype B virus in Hong Kong MSM population. The understanding of transmission and growth model of the subtype B epidemic provides more information on the HIV-1 transmission among MSM population in other Asia Pacific high-income countries.

HIV-1 group M strains are characterized into 9 pure subtypes and 48 circulating recombinant forms (CRFs). Recent studies have identified the presence of new HIV-1 recombinants in Hong Kong and their complexity continues to increase. This study aims to characterize the HIV-1 genetic diversity in Hong Kong. Phylogenetic analyses were performed by using HIV-1 pol sequences including protease and partial reverse transcriptase isolated from 1045 local patients in Hong Kong from 2003 to 2008. For the pol sequences with unassigned genotype, the evidence of recombination was determined by using sliding-window based bootscan plots and their env C2V3 region were also sequenced. Epidemiological background of these patients was further collected. The pol phylogenetic analyses highlighted the extent of HIV-1 genetic diversity in Hong Kong. Subtype B (450/1045; 43.1%) and CRF01_AE (469/1045; 44.9%) variants were clearly predominant. Other genotypes (126/1045; 12.1%) including 3 defined subtypes, 10 CRFs, 1 unassigned subtype and 33 recombinants with 11 different mosaic patterns were observed. Recombinants of subtype B and CRF01_AE were mainly found among local Chinese MSM throughout 2004 to 2008, while the CRF02_AG and subtype G recombinants were circulating among non-Chinese Asian population in Hong Kong through heterosexual transmission starting from 2008. Our study demonstrated the complex recombination of HIV-1 in Hong Kong and the need in developing surveillance system for tracking the distribution of new HIV-1 genetic variants.

Due to the refractiveness of tumor tissues to adeno-associated virus (AAV) transduction, AAV vectors are poorly explored for cancer therapy delivery. Here, we aimed to engineer AAVs to target tumors by enabling the specific engagement of fibroblast activation protein (FAP). FAP is a cell surface receptor distinctly upregulated in the reactive tumor stroma, but rarely expressed in healthy tissues. Thus, targeting FAP presents an opportunity to selectively transduce tumor tissues. To achieve this, we modified the capsid surface of AAV2 with an αFAP nanobody to retarget the capsid to engage FAP receptor. Following transduction, we observed a 23- to 80-fold increase in the selective transduction of FAP + tumor cells in vitro , and greater than 5-fold transduction of FAP + tumor tissues in vivo. Subsequent optimization of the VP1-nanobody expression cassette further enhanced the transduction efficiency of the modified capsids. Due to the limited αFAP nanobodies repertoires, we broadened the versatility of this high-fidelity platform by screening a naive VHH yeast display library, leading to the identification of several novel αFAP nanobody candidates (K D  = 0.1 to >100 nM). Hence, our study offers new opportunity for the application of AAV vectors for highly selective delivery of therapeutics to the tumor stroma., Competing Interests: O.O., F.H., P.M., U.M., and F.d.P. are employees of Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany. P.C., C.K., and T.P. are employees of Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, USA., (© 2024 The Authors.)

Discovery and optimization of a biotherapeutic monoclonal antibody requires a careful balance of target engagement and physicochemical developability properties. To take full advantage of the sequence diversity provided by different antibody discovery platforms, a rapid and reliable process for humanization of antibodies from nonhuman sources is required. Canonically, maximizing homology of the human variable region (V-region) to the original germline was believed to result in preservation of binding, often without much consideration for inherent molecular properties. We expand on this approach by grafting the complementary determining regions (CDRs) of a mouse anti-LAG3 antibody into an extensive matrix of human variable heavy chain (VH) and variable light chain (VL) framework regions with substantially broader sequence homology to assess the impact on complementary determining region-framework compatibility through progressive evaluation of expression, affinity, biophysical developability, and function. Specific VH and VL framework sequences were associated with major expression and purification phenotypes. Greater VL sequence conservation was correlated with retained or improved affinity. Analysis of grafts that bound the target demonstrated that initial developability criteria were significantly impacted by VH, but not VL. In contrast, cell binding and functional characteristics were significantly impacted by VL, but not VH. Principal component analysis of all factors identified multiple grafts that exhibited more favorable antibody properties, notably with nonoptimal sequence conservation. Overall, this study demonstrates that modern throughput systems enable a more thorough, customizable, and systematic analysis of graft-framework combinations, resulting in humanized antibodies with improved global properties that may progress through development more quickly and with a greater probability of success., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Purpose: Semaphorin 3A (Sema3A) is an axonal guidance molecule that inhibits angiogenesis by vasorepulsion and blocks revascularization in the ischemic retina. BI-X is an intravitreal anti-Sema3A agent under clinical investigation in patients with proliferative diabetic retinopathy (PDR) and diabetic macular ischemia (DMI)., Methods: Surface plasmon resonance was used to determine binding affinity of BI-X to human and murine Sema3A. In vitro, human retinal microvascular endothelial cells (HRMECs) were used to assess effects of BI-X on cell permeability and cytoskeletal collapse induced by Sema3A. In vivo, intravitreal BI-X or an anti-trinitrophenol control antibody was administered in both eyes in mice with oxygen-induced retinopathy (OIR). Retinal flat mounts were prepared, and avascular area and tip cell density were determined using confocal laser-scanning microscopy., Results: Dissociation constants for BI-X binding to human and murine Sema3A were 29 pM and 27 pM, respectively. In vitro, BI-X prevented HRMEC permeability and cytoskeletal collapse induced by Sema3A. In vivo, BI-X increased tip cell density by 33% (P < 0.001) and reduced avascular area by 12% (not significant). A significant negative correlation was evident between avascular area and tip cell density (r2 = 0.4205, P < 0.0001)., Conclusions: BI-X binds to human Sema3A with picomolar affinity and prevents cell permeability and cytoskeletal collapse in HRMECs. BI-X also enhances revascularization in mice with OIR., Translational Relevance: BI-X is a potent inhibitor of human Sema3A that improves revascularization in a murine model of OIR; BI-X is currently being investigated in patients with laser-treated PDR and DMI.

Aim: The fully automated microfluidics-based Gyrolab is a popular instrument for the bioanalysis of protein therapeutics; requiring minimal sample and reagent volumes. Gyros offers affinity software for determining binding affinity in solution using a high-throughput method and miniaturized reactions., Results: Using this affinity software, multiple CTGF-targeting reagents were characterized on the Gyrolab after <100% target coverage was seen in a cynomolgus pharmacokinetic/PD study dosed with anti-CTGF antibodies. The results uncovered magnitude differences in binding affinities between the dosed antibody, target and assay reagents., Conclusion: The binding affinity values were used to investigate reduced target coverage and results highlight potential of the affinity software for incorporation into the bioanalyst's existing Gyrolab workflow for characterizing reagents and optimizing pharmacokinetic/PD bioanalytical assays.

Therapeutic agents antagonizing B-cell-activating factor/B-lymphocyte stimulator (BAFF/BLyS) are currently in clinical development for autoimmune diseases; belimumab is the first Food and Drug Administration-approved drug in more than 50 years for the treatment of lupus. As a member of the tumor necrosis factor superfamily, BAFF promotes B-cell survival and homeostasis and is overexpressed in patients with systemic lupus erythematosus and other autoimmune diseases. BAFF exists in three recognized forms: membrane-bound and two secreted, soluble forms of either trimeric or 60-mer oligomeric states. To date, most in vitro pharmacology studies of BAFF neglect one or more of these forms. Here, we report a comprehensive in vitro cell-based analysis of BAFF in assay systems that measure all forms of BAFF-mediated activation. We demonstrate the effects of these BAFF forms in both a primary human B-cell proliferation assay and in nuclear factor κB reporter assay systems in Chinese hamster ovary cells expressing BAFF receptors and transmembrane activator and calcium-modulator and cyclophilin ligand interactor (TACI). In contrast to the mouse system, we find that BAFF trimer activates the human TACI receptor. Further, we profiled the activities of two clinically advanced BAFF antagonist antibodies, belimumab and tabalumab. Unexpectedly, we revealed differences in inhibitory potencies against the various BAFF forms, in particular that belimumab does not potently inhibit BAFF 60-mer. Through this increased understanding of the activity of BAFF antagonists against different forms of BAFF, we hope to influence the discovery of BAFF antagonist antibodies with distinct therapeutic mechanisms for improvement in the treatment of lupus or other related autoimmune pathologies., (Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.)

Insulin-like growth factor receptor (IGF-1R) is a growth factor receptor tyrosine kinase that acts as a critical mediator of cell proliferation and survival. Inhibitors of this receptor are believed to provide a new target in cancer therapy. We previously reported an isoquinolinedione series of IGF-1R inhibitors. Now we have identified a series of 3-cyanoquinoline compounds that are low nanomolar inhibitors of IGF-1R. The strategies, synthesis, and SAR behind the cyanoquinoline scaffold will be discussed.

Insulin-like growth factor receptor (IGF-1R) is a growth factor receptor tyrosine kinase that acts as a critical mediator of cell proliferation and survival. This receptor is over-expressed or activated in tumor cells and is emerging as a novel target in cancer therapy. Efforts in our "Hit to Lead" group have generated a novel series of submicromolar IGF-1R inhibitors based on a isoquinolinedione template originating from a Lance enzyme HTS screen. Chemical triage and parallel synthesis incorporating focused library arrays were instrumental in moving these investigations through the Wyeth exploratory medicinal chemistry process. The strategies, synthesis, and SAR behind this interesting kinase scaffold will be described.

ZipA is a membrane anchored protein in Escherichia coli that interacts with FtsZ, a homolog of eukaryotic tubulins, forming a septal ring structure that mediates bacterial cell division. Thus, the ZipA/FtsZ protein-protein interaction is a potential target for an antibacterial agent. We report here an NMR-based fragment screening approach which identified several hits that bind to the C-terminal region of ZipA. The screen was performed by 1H-15N HSQC experiments on a library of 825 fragments that are small, lead-like, and highly soluble. Seven hits were identified, and the binding mode of the best one was revealed in the X-ray crystal structure. Similar to the ZipA/FtsZ contacts, the driving force in the binding of the small molecule ligands to ZipA is achieved through hydrophobic interactions. Analogs of this hit were also evaluated by NMR and X-ray crystal structures of these analogs with ZipA were obtained, providing structural information to help guide the medicinal chemistry efforts.

The ZipA-FtsZ protein-protein interaction is a potential target for antibacterial therapy. The design and parallel synthesis of a combinatorial library of small molecules, which target the FtsZ binding area on ZipA are described. Compounds were demonstrated to bind to the FtsZ binding domain of ZipA by HSQC NMR and to inhibit cell division in a cell elongation assay.

The binding of FtsZ to ZipA is a potential target for antibacterial therapy. Based on a small molecule inhibitor of the ZipA-FtsZ interaction, a parallel synthesis of small molecules was initiated which targeted a key region of ZipA involved in FtsZ binding. The X-ray crystal structure of one of these molecules complexed with ZipA was solved. The structure revealed an unexpected binding mode, facilitated by desolvation of a loosely bound surface water.

A fluorescence polarization competition assay has been developed to screen for inhibitors of the Escherichia coli FtsZ/ZipA protein-protein interaction. A previously published X-ray costructure demonstrated that a 17-amino-acid peptide, corresponding to FtsZ C-terminal residues 367-383 (FtsZ(367-383)), interacts with the C-terminal FtsZ binding domain of ZipA (ZipA(185-328)). Phage display was employed to identify a unique but related peptide which when further modified and labeled was shown to have a higher affinity to ZipA(185-328) than the FtsZ(367-383) peptide and binds to the same site. This peptide had a six fold increase in fluorescence polarization upon binding to ZipA(185-328) compared to a two fold increase for the FtsZ(367-383) fluorophore. As a result, assay parameters using the phage display peptide were further optimized and adapted for the high-throughput screen. A high-throughput screen of 250,000 compounds identified 29 hits with inhibition equal to or greater than 30% at 50 microg/ml. An X-ray costructure of a promising small molecule in this library complexed with ZipA(185-328) (KI=12 microM) revealed that the compound binds to the same hydrophobic pocket as the FtsZ(367-383) peptide.