Cardiopulmonary Exercise Testing in Children and Adolescents with Repaired Tetralogy of Fallot: Mechanisms of Exercise Intolerance and Clinical Implications.

By comparison with adults, cardiopulmonary exercise testing in children with Tetralogy of Fallot is limited, and its clinical application less clarified. This study provides a comprehensive CPET profile in a child-adolescent population with repaired TOF, explores mechanisms underpinning exercise intolerance and associations with clinical outcome. Seventy-four CPETs were completed in 58 child-adolescents with rTOF (age 13.8 SD 2.4 years). CPET parameters were corrected for age, sex and body size. At follow-up (4.9 years, IQR 3.5-7.9) clinical status and re-intervention was evaluated and CPET indices predicting these outcomes determined. Cohort peak V̇O2 was within low-normal limits (% pred: 74.1% SD 15.4) with 15 patients (26%) displaying moderately severe reduction in V̇O2peak (< 65% pred). Oxygen uptake efficiency slope highly correlated with V̇O2peak (r = 0.94, p < 0.001) and was insensitive to exercise intensity. No significant change in CPET occurred in patients who underwent interval testing at 24 SD 14.5 months, although there was a variable response in V̇O2peak between individuals. Chronotropic response, lung vital capacity, heart rate-V̇O2 slope (indicator of stroke volume) predicted oxygen consumption: V̇O2peak (R2 = 50.91%, p < 0.001) and workload (R2 = 58.39%, p < 0.001). Adverse clinical status was associated with reduced workload (OR 0.97, p = 0.011). V̇E/V̇CO2 slope was steeper in those that died ((%pred:137.8 SD 60.5 vs. 108.4 SD 17.0, p < 0.019). RVOT reintervention post-CPET (24 patients, 43.8%) was associated with an increased gradient of HR-VO2 slope (OR 1.042, p = 0.004). In child-adolescents with TOF important reductions in cardiopulmonary functioning were apparent in 25% of patients. Exercise intolerance was related to reduced vital capacity, impaired chronotropic response and deficient stroke volume increment.

CryoEM and AI reveal a structure of SARS-CoV-2 Nsp2, a multifunctional protein involved in key host processes.

The SARS-CoV-2 protein Nsp2 has been implicated in a wide range of viral processes, but its exact functions, and the structural basis of those functions, remain unknown. Here, we report an atomic model for full-length Nsp2 obtained by combining cryo-electron microscopy with deep learning-based structure prediction from AlphaFold2. The resulting structure reveals a highly-conserved zinc ion-binding site, suggesting a role for Nsp2 in RNA binding. Mapping emerging mutations from variants of SARS-CoV-2 on the resulting structure shows potential host-Nsp2 interaction regions. Using structural analysis together with affinity tagged purification mass spectrometry experiments, we identify Nsp2 mutants that are unable to interact with the actin-nucleation-promoting WASH protein complex or with GIGYF2, an inhibitor of translation initiation and modulator of ribosome-associated quality control. Our work suggests a potential role of Nsp2 in linking viral transcription within the viral replication-transcription complexes (RTC) to the translation initiation of the viral message. Collectively, the structure reported here, combined with mutant interaction mapping, provides a foundation for functional studies of this evolutionary conserved coronavirus protein and may assist future drug design.

Fragment binding to the Nsp3 macrodomain of SARS-CoV-2 identified through crystallographic screening and computational docking

The SARS-CoV-2 macrodomain (Mac1) within the non-structural protein 3 (Nsp3) counteracts host-mediated antiviral ADP-ribosylation signalling. This enzyme is a promising antiviral target because catalytic mutations render viruses non-pathogenic. Here, we report a massive crystallographic screening and computational docking effort, identifying new chemical matter primarily targeting the active site of the macrodomain. Crystallographic screening of diverse fragment libraries resulted in 214 unique macrodomain-binding fragments, out of 2,683 screened. An additional 60 molecules were selected from docking over 20 million fragments, of which 20 were crystallographically confirmed. X-ray data collection to ultra-high resolution and at physiological temperature enabled assessment of the conformational heterogeneity around the active site. Several crystallographic and docking fragment hits were validated for solution binding using three biophysical techniques (DSF, HTRF, ITC). Overall, the 234 fragment structures presented explore a wide range of chemotypes and provide starting points for development of potent SARS-CoV-2 macrodomain inhibitors.

The use of a co-design model in improving timely bleed reporting by adults with haemophilia living in the Auckland region of New Zealand.

Many adult patients diagnosed with phenotypically moderate and severe haemophilia living in the Auckland region of New Zealand do not report bleeding episodes within a timeframe that allows for optimal assessment and management. This can result in poor clinical outcomes for patients and poor oversight of the use of expensive clotting factor concentrates. Our goal was to improve both the number and speed at which bleeding episodes were reported to our centre, improving access to care and clinical oversight of the use of expensive factor concentrates and aiding the development of a care partnership with patients. We worked with 70 adult PWH living in the Auckland region of New Zealand with moderate and severe haemophilia A and B. Over a 5-month period between March and July 2013 we used a co-design model to develop and implement a range of strategies to improve the timing and frequency of bleed reporting. Mean bleed reporting time was reduced threefold, with a threefold increase in the number of bleeds reported per month. We reduced the number of bleeding episodes reported outside of a prespecified 48-h time limit by 68%. We significantly improved bleed reporting and time to report, indicating improved access to our services, improved clinical oversight and improved accountability to our national funder. We have achieved a care partnership and a reduction in factor consumption for the study population without compromising the quality of care they receive.

Conservative physiotherapeutic management of chronic haematomata and haemophilic pseudotumours: case study and comparison to historical management.

A conservative, non-operative physiotherapeutic regime for the management of chronic haematomata and pseudotumours in patients suffering from haemophilia is described in this article. Two cases are described where physiotherapy treatment is applied to large masses at the shoulder and femur respectively, where therapy commenced within the first 6 months following onset. These are presented relative to a case that was managed over a much longer period without early physiotherapy input, and the relative outcomes are examined. While both the early physiotherapy-managed cases showed a complete resolution at follow-up examination, the more established chronic pseudotumour required surgical excision, with significant residual muscle contractility, length and strength issues noted on clinical and magnetic resonance imaging reviews. No adverse symptoms or haemostatic issues were reported in response to this less invasive treatment regime by either patient in the two conservative physiotherapy cases.

Domiciliary application of CryoCuff in severe haemophilia: qualitative questionnaire and clinical audit.

The acute management of haemophilic bleeding episodes in the home setting is based on the concept of immediate factor replacement therapy and the PRICE regime--an acronym representing the concepts of Protection, Rest, Ice, Compression and Elevation [1,2]. Integral to this regime is the application of cold therapy, and yet little is known regarding the safe periods of application, or the relative safety of cryotherapy devices such as the CryoCuff when used in the home setting by patients suffering from severe haemophilia and related bleeding disorders. This study examines the subjective patient response to the application of the CryoCuff device in the home setting in terms of the effect on pain, joint swelling and the return to 'pre-bleed status' of the knee, ankle or elbow in patients with severe haemophilia A/B or type III von Willebrand's disease (VWD) immediately following haemarthrosis, and any potential adverse effects related to the device or recommended duration of application as stated in the PRICE guideline (Fig. 1). Twelve patients, either with severe haemophilia A/B or with VWD were recruited and asked to use the CryoCuff device as part of the PRICE regime immediately following the onset of knee-, ankle- or elbow bleeds for the next one year. Each subject was then sent a qualitative questionnaire to determine subjective responses to the device. All patients reported that the application protocol was easy to follow, they were able to apply the device as per the PRICE regime and they were able to tolerate it for the recommended period. Whereas, all the patients felt that the device had a significant impact on alleviation of pain and return to pre-bleed status, 78% of the patients felt that the device led to a significant reduction in swelling around the affected joint. There was no conclusive evidence that the device resulted in any reduction in the amount of factor used to treat the acute bleeding episode, however, no patients reported any perceived delay in achieving haemostasis or required extra factor replacement therapy consequent to the usage of the device. No other adverse effects were reported by participants in this study.

Detection of TMPRSS2:ERG fusion gene in circulating prostate cancer cells / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To investigate the existence of TMPRSS2:ERG fusion gene in circulating tumor cells (CTC) from prostate cancer patients and its potential in monitoring tumor metastasis.</p><p><b>METHODS</b>We analyzed the frequency of TMPRSS2:ERG and TMPRSS2:ETV1 transcripts in 27 prostate cancer biopsies from prostatectomies, and TMPRSS2:ERG transcripts in CTC isolated from 15 patients with advanced androgen independent disease using reverse transcription polymerase chain reaction (RT-PCR). Fluorescence in situ hybridization (FISH) was applied to analyze the genomic truncation of ERG, which is the result of TMPRSS2:ERG fusion in 10 of the 15 CTC samples.</p><p><b>RESULTS</b>TMPRSS2:ERG transcripts were found in 44% of our samples, but we did not detect expression of TMPRSS2:ETV1. Using FISH analysis we detected chromosomal rearrangements affecting the ERG gene in 6 of 10 CTC samples, including 1 case with associated TMPRSS2:ERG fusion at the primary site. However, TMPRSS2:ERG transcripts were not detected in any of the 15 CTC samples, including the 10 cases analyzed by FISH.</p><p><b>CONCLUSION</b>Although further study is required to address the association between TMPRSS2:ERG fusion and prostate cancer metastasis, detection of genomic truncation of the ERG gene by FISH analysis could be useful for monitoring the appearance of CTC and the potential for prostate cancer metastasis.</p>