PDXK mutations cause polyneuropathy responsive to pyridoxal 5'-phosphate supplementation.

OBJECTIVE: To identify disease-causing variants in autosomal recessive axonal polyneuropathy with optic atrophy and provide targeted replacement therapy. METHODS: We performed genome-wide sequencing, homozygosity mapping, and segregation analysis for novel disease-causing gene discovery. We used circular dichroism to show secondary structure changes and isothermal titration calorimetry to investigate the impact of variants on adenosine triphosphate (ATP) binding. Pathogenicity was further supported by enzymatic assays and mass spectroscopy on recombinant protein, patient-derived fibroblasts, plasma, and erythrocytes. Response to supplementation was measured with clinical validated rating scales, electrophysiology, and biochemical quantification. RESULTS: We identified biallelic mutations in PDXK in 5 individuals from 2 unrelated families with primary axonal polyneuropathy and optic atrophy. The natural history of this disorder suggests that untreated, affected individuals become wheelchair-bound and blind. We identified conformational rearrangement in the mutant enzyme around the ATP-binding pocket. Low PDXK ATP binding resulted in decreased erythrocyte PDXK activity and low pyridoxal 5'-phosphate (PLP) concentrations. We rescued the clinical and biochemical profile with PLP supplementation in 1 family, improvement in power, pain, and fatigue contributing to patients regaining their ability to walk independently during the first year of PLP normalization. INTERPRETATION: We show that mutations in PDXK cause autosomal recessive axonal peripheral polyneuropathy leading to disease via reduced PDXK enzymatic activity and low PLP. We show that the biochemical profile can be rescued with PLP supplementation associated with clinical improvement. As B6 is a cofactor in diverse essential biological pathways, our findings may have direct implications for neuropathies of unknown etiology characterized by reduced PLP levels. ANN NEUROL 2019;86:225-240

The Aresys FF-SAR Service for Cryosat-2 at ESA GPOD

The Fully-Focused SAR (FF-SAR) processing, introduced in Egido and Smith (2016) allows obtaining a maximum resolution of 0.5 m in the along-track direction. It provides significant benefits for inland water altimetry investigations allowing the successful investigation of very small rivers and canals (Kleinherenbrink, 2020) that are typically harder to be analysed by using unfocused Delay-Doppler SAR (DD-SAR) data (about 300 m resolution in the along-track direction).In its development, two major limitations were associated with the FF-SAR processing: 1) the presence of evenly spaced high sidelobes in the Point Target Response (PTR) due to the closed-loop burst mode implemented in Sentinel-3 & Cryosat-2 altimeter payloads, used for initial FF-SAR investigations, and 2) the heavy computational burden with respect to the unfocused DD-SAR processing. The first limitation can be overcome by designing the radar system differently adopting an open-loop transmission scheme as, for instance, the one implemented in the altimeter payload of the Sentinel-6 Michael Freilich mission, launched on 21 November 2020. The second limitation has been addressed in research works following Egido and Smith (2016) indicating that an improvement in terms of computational burden can be achieved by adopting algorithms in the frequency domain (Guccione et al., 2018).Being the role of FF-SAR for future inland water altimetry well understood, along with the possibility to see it implemented with reduced drawbacks during the Sentinel-6 Michael Freilich mission, a collaboration has started between the ESA GPOD Team, already hosting the successful SARvatore services portfolio for unfocused SAR & SARin altimetry, and Aresys. Aresys has developed a generic FF-SAR prototype processor, that is able to process data acquisition from different instruments and exploiting the frequency-domain Omega-K algorithm (Guccione et al., 2018 & Scagliola et al., 2018). The Aresys's FFSAR prototype processor for CryoSat-2 allows users to process, on line and on demand, low-level CryoSat FBR products in SAR mode up to FF-SAR Level-1 products with self-customized options. Additionally a wide set of processing parameters is configurable, allowing as an example to select the along-track resolution or to obtain FFSAR multilooked waveforms at the desired posting rate. The collaboration led to the creation of a new service for the processing of CryoSat Baseline D data in FF-SAR mode. Users will be able to select the following options: 1) range oversampling factor, 2) bandwidth factor (responsible for the along-track resolution value) and 3) multilook posting rate (1Hz-500Hz). Geophysical corrections and L2 estimates from both a threshold peak retracker and an ALES-like subwaveform retracker are part of the output package. In preliminary open ocean analyses, very good results on SSH noise have been obtained by the ALES-like subwaveform retracker.In this presentation, the Aresys FF-SAR prototype processor is described and the outcome of some preliminary validation activities, performed by a group of altimetry researchers, is reported. The service is scheduled to open to all GPOD/SARvatore users in the first semester of 2021. Future evolutions should include the extension of the service to Sentinel-3 and Sentinel-6 data.Astrodynamics & Space Mission

Cyclin D-CDK4 Disulfide Bond Attenuates Pulmonary Vascular Cell Proliferation.

BACKGROUND: Pulmonary hypertension (PH) is a chronic vascular disease characterized, among other abnormalities, by hyperproliferative smooth muscle cells and a perturbed cellular redox and metabolic balance. Oxidants induce cell cycle arrest to halt proliferation; however, little is known about the redox-regulated effector proteins that mediate these processes. Here, we report a novel kinase-inhibitory disulfide bond in cyclin D-CDK4 (cyclin-dependent kinase 4) and investigate its role in cell proliferation and PH. METHODS: Oxidative modifications of cyclin D-CDK4 were detected in human pulmonary arterial smooth muscle cells and human pulmonary arterial endothelial cells. Site-directed mutagenesis, tandem mass-spectrometry, cell-based experiments, in vitro kinase activity assays, in silico structural modeling, and a novel redox-dead constitutive knock-in mouse were utilized to investigate the nature and definitively establish the importance of CDK4 cysteine modification in pulmonary vascular cell proliferation. Furthermore, the cyclin D-CDK4 oxidation was assessed in vivo in the pulmonary arteries and isolated human pulmonary arterial smooth muscle cells of patients with pulmonary arterial hypertension and in 3 preclinical models of PH. RESULTS: Cyclin D-CDK4 forms a reversible oxidant-induced heterodimeric disulfide dimer between C7/8 and C135, respectively, in cells in vitro and in pulmonary arteries in vivo to inhibit cyclin D-CDK4 kinase activity, decrease Rb (retinoblastoma) protein phosphorylation, and induce cell cycle arrest. Mutation of CDK4 C135 causes a kinase-impaired phenotype, which decreases cell proliferation rate and alleviates disease phenotype in an experimental mouse PH model, suggesting this cysteine is indispensable for cyclin D-CDK4 kinase activity. Pulmonary arteries and human pulmonary arterial smooth muscle cells from patients with pulmonary arterial hypertension display a decreased level of CDK4 disulfide, consistent with CDK4 being hyperactive in human pulmonary arterial hypertension. Furthermore, auranofin treatment, which induces the cyclin D-CDK4 disulfide, attenuates disease severity in experimental PH models by mitigating pulmonary vascular remodeling. CONCLUSIONS: A novel disulfide bond in cyclin D-CDK4 acts as a rapid switch to inhibit kinase activity and halt cell proliferation. This oxidative modification forms at a critical cysteine residue, which is unique to CDK4, offering the potential for the design of a selective covalent inhibitor predicted to be beneficial in PH

A New Fully-Focused SAR Altimetry Processor in the ESA G-POD SARvatore Family: Validation and Applications on Inland Waters

Satellite altimetry has successfully been applied to derive water level time series over lakes and rivers for many years. In the last decade major advancement within the field have been achieved. The SAR altimetry era, initiated with the launch of CryoSat-2, has made it possible to measure much smaller targets more accurately. The standard 20 Hz radar altimetry products generally allow to study targets down to a few hundred meters in optimal conditions. However, recent advancement in the low-level data processing, the Fully-Focused SAR (FFSAR) technique, where the footprint in the along-track direction can be as small as a few meters, has made it possible to measure the water level of even smaller targets. FFSAR processed data from CryoSat-2 in SAR mode is made freely available by ESA through a newly integrated service on the Grid Processing on Demand (G-POD) platform, within the SARvatore Family of custom processors, where the user can order data from an area and specify various parameters to be applied in the processing. The FFSAR processor has been developed within the ESA ESTEC Sentinel-6 Project and adapted to CryoSat-2 for verification and validation purposes. Here we demonstrate the value of FFSAR and evaluate the FFSAR product obtained from the ESA G-POD service over different inland water targets. The evaluation and validation is done via in situ water level data, external processed FFSAR data, unfocused SAR data and laser altimetry data from ICESat-2. More specific, we perform an evaluation over the narrow ( < 100 m) American Rivers: Red River, Little River, and Canadian River located in the Mississippi basin. Where we compare the water level based on different retracker e.g. ALES+ and threshold retrackers. We make a joint water level time series by considering data from CryoSat-2 and ICESat-2 over a reach, and validate the results against in situ data. Preliminary results show that we can capture the main part of the water level signal. Over the Elbe River, we compare the FFSAR based river water levels with results based on unfocused SAR processing and in situ gauges. We extend our analysis with a case study conducted over the Lake IJssel which is located in The Netherlands and covers an area of approximately 1100 km2. We examine the performance of the G-POD processor by evaluating the FFSAR lake height estimates using externally processed FFSAR data that have been validated against in situ data.Astrodynamics & Space Mission