Search

Purpose Radioiodine I-131 (RAI) is the therapy of choice for differentiated thyroid cancer (DTC). Between 5% and 15% of DTC patients become RAI refractory, due to the loss of expression/function of iodide metabolism components, especially the Na/I symporter (NIS). We searched for a miRNA profile associated with RAI-refractory DTC to identify novel biomarkers that could be potential targets for redifferentiation therapy. Methods We analyzed the expression of 754 miRNAs in 26 DTC tissues: 12 responsive (R) and 14 non-responsive (NR) to RAI therapy. We identified 15 dysregulated miRNAs: 14 were upregulated, while only one (miR-139-5p) was downregulated in NR vs. R tumors. We investigated the role of miR-139-5p in iodine uptake metabolism. We overexpressed miR-139-5p in two primary and five immortalized thyroid cancer cell lines, and we analyzed the transcript and protein levels of NIS and its activation through iodine uptake assay and subcellular protein localization. Results The finding of higher intracellular iodine levels and increased cell membrane protein localization in miR-139-5p overexpressing cells supports the role of this miRNA in the regulation of NIS function. Conclusions Our study provides evidence of miR-139-5p involvement in iodine uptake metabolism and suggests its possible role as a therapeutic target in restoring iodine uptake in RAI-refractory DTC.

The weighted ensemble (WE) family of methods is one of several statistical-mechanics based path sampling strategies that can provide estimates of key observables (rate constants, pathways) using a fraction of the time required by direct simulation methods such as molecular dynamics or discrete-state stochastic algorithms. WE methods oversee numerous parallel trajectories using intermittent overhead operations at fixed time intervals, enabling facile interoperability with any dynamics engine. Here, we report on major upgrades to the WESTPA software package, an open-source, high-performance framework that implements both basic and recently developed WE methods. These upgrades offer substantial improvements over traditional WE. Key features of the new WESTPA 2.0 software enhance efficiency and ease of use: an adaptive binning scheme for more efficient surmounting of large free energy barriers, streamlined handling of large simulation datasets, exponentially improved analysis of kinetics, and developer-friendly tools for creating new WE methods, including a Python API and resampler module for implementing both binned and “binless” WE strategies.Table of Contents/Abstract ImageFor the manuscript “WESTPA 2.0: High-performance upgrades for weighted ensemble simulations and analysis of longer-timescale applications” by Russo et al.

Seq-Well is a high-throughput, picowell-based single-cell RNA-seq technology that can be used to simultaneously profile the transcriptomes of thousands of cells (Gierahn et al. Nat Methods 14(4):395-398, 2017). Relative to its reverse-emulsion-droplet-based counterparts, Seq-Well addresses key cost, portability, and scalability limitations. Recently, we introduced an improved molecular biology for Seq-Well to enhance the information content that can be captured from individual cells using the platform. This update, which we call Seq-Well S

PURPOSE To retrospectively estimate the impact of radiotherapy as a progression-directed therapy (PDT) in oligoprogressive metastatic castration-resistant prostate cancer (mCRPC) patients under androgen receptor-target therapy (ARTT). MATERIALS AND METHODS mCRPC patients are treated with PDT. End-points were time to next-line systemic treatment (NEST), radiological progression-free survival (r-PFS) and overall survival (OS). Toxicity was registered according to Common Terminology Criteria for Adverse Events v4.0. Survival analysis was performed using the Kaplan-Meier method; univariate and multivariate analyses were performed. RESULTS Fifty-seven patients were analyzed. The median follow-up after PDT was 25.2 months (interquartile, 17.1-44.5). One-year NEST-free survival, r-PFS and OS were 49.8%, 50.4% and 82.1%, respectively. At multivariate analysis, polymetastatic condition at diagnosis of metastatic hormone-sensitive prostate cancer (mHSPC) (HR 2.82, p = 0.004) and PSA doubling time at diagnosis of mCRPC (HR 2.76, p = 0.006) were associated with NEST-free survival. The same variables were associated with r-PFS (HR 2.32, p = 0.021; HR 2.24, p = 0.021). One patient developed late grade ≥ 2 toxicity. CONCLUSION Our study shows that radiotherapy in oligoprogressive mCRPC is safe, is effective and seems to prolong the efficacy of ARTT in patients who otherwise would have gone systemic treatment switch, positively affecting disease progression. Prospective trials are needed.

We present six advanced tutorials instructing users in the best practices of using key new features and plugins/extensions of the WESTPA 2.0 software package, which consists of major upgrades for enabling applications of the weighted ensemble (WE) path sampling strategy to even larger systems and/or slower processes. The tutorials demonstrate the use of the following key features: (i) a generalized resampler module for the creation of “binless” schemes, (ii) a minimal adaptive binning scheme for more efficient surmounting of free energy barriers, (iii) streamlined handling of large simulation datasets using an HDF5 framework, (iv) two different schemes for more efficient rate-constant estimation, (v) a Python API for simplified analysis of WE simulations, and (vi) plugins/extensions for Markovian Weighted Ensemble Milestoning and WE rule-based modeling at the system biology level. Applications of the tutorials range from atomistic to residue-level to non-spatial models, and include complex processes such as protein folding and the membrane permeability of a drug-like molecule. Users are expected to already have significant experience with running conventional molecular dynamics simulations and completed the previous suite of WESTPA tutorials.

Formates are promising salts for hydrogen storage. They can be catalytically converted to bicarbonate at near-ambient conditions, and regenerated under moderate pressure for H2 uptake. Moreover, the system is completely safe, non-toxic, and easy to handle. Up to date, a few heterogeneous catalytic systems have been proposed to carry this transformation. However, many criticisms still have to be addressed, including reaction kinetics for high power applications, stability, cyclability, and supersaturated solutions to increase energetic density. In this work, a critical review of the state of art of such system is presented, highlighting theoretical limitations and applicative shortcomings, and giving perspective on critical issues to be addressed in future research. Despite the actual bottlenecks, the system is still scarcely explored and there is promising room for improvement. State-of-the-art catalytic systems could provide 72% energetic efficiency, that could be improved up to 90%.

The worldwide coronavirus pandemic in 2020 radically changed the landscape of psychology service provision and training, with rapid rollouts of telemedicine to promote safe access to care. In this perspective article, we share the experiences of 4 psychology trainees, all of whom worked as psychology interns or postdoctoral fellows in pediatric behavioral sleep medicine during the pandemic. With restricted in-person visits and upheaval of children’s normative sleep and school schedules, we directly observed growth in both need for psychological care and opportunity to provide this care remotely. Here, we summarize the unique challenges and learning opportunities we faced when trying to learn and implement evidence-based assessment and treatment of child and adolescent sleep difficulties during the pandemic. CITATION: McQuillan ME, Anderson A, Russo KD, Truss A, Honaker SM, Walsh KL. Pediatric behavioral sleep medicine in the era of telemedicine: psychology trainee perspectives. J Clin Sleep Med. 2021;17(9):1965–1967.

Regulation of pancreatic K

Regulation of pancreatic KATP channels involves orchestrated interactions of channel subunits, Kir6.2 and SUR1, and their ligands. How ligand interactions affect channel conformations and activity is not well understood. To elucidate the structural correlates pertinent to ligand interactions and channel gating, we compared cryo-EM structures of channels in the presence and absence of pharmacological inhibitors and ATP, focusing on channel conformational dynamics. We found pharmacological inhibitors and ATP enrich a channel conformation in which the Kir6.2 cytoplasmic domain is closely associated with the transmembrane domain relative to one where the Kir6.2 cytoplasmic domain is extended away into the cytoplasm. This conformation change remodels a network of intra and inter-subunit interactions as well as both the ATP and PIP2 binding pockets. The structures resolved key contacts between the distal N-terminus of Kir6.2 and SUR1’s ABC module involving residues implicated in channel function. A SUR1 residue, K134, is identified to directly contribute to the PIP2 binding pocket. Molecular dynamics simulations revealed two Kir6.2 residues, K39 and R54, that mediate both ATP and PIP2 binding, suggesting a mechanism for competitive gating by ATP and PIP2.

In brief Proper development of ovarian follicles, comprised of an oocyte and surrounding somatic cells, is essential to support female fertility and endocrine health. Here, we describe a method to isolate single oocytes and somatic cells from the earliest stage follicles, called primordial follicles, and we characterize signals that drive their activation. Abstract Primordial follicles are the first class of follicles formed in the mammalian ovary and are comprised of an oocyte surrounded by a layer of squamous pre-granulosa cells. This developmental class remains in a non-growing state until individual follicles activate to initiate folliculogenesis. What regulates the timing of follicle activation and the upstream signals that govern these processes are major unanswered questions in ovarian biology. This is partly due to the paucity of data on staged follicle cells since isolating and manipulating individual oocytes and somatic cells from early follicle stages are challenging. To date, most studies on isolated primordial follicles have been conducted on cells collected from animal-age- or oocyte size-specific samples, which encompass multiple follicular stages. Here, we report a method for collecting primordial follicles and their associated oocytes and somatic cells from neonatal murine ovaries using liberase, DNase I, and Accutase. This methodology allows for the identification and collection of follicles immediately post-activation enabling unprecedented interrogation of the primordial-to-primary follicle transition. Molecular profiling by single-cell RNA sequencing revealed that processes including organelle disassembly and cadherin binding were enriched in oocytes and somatic cells as they transitioned from primordial to the primary follicle stage. Furthermore, targets including WNT4, TGFB1, FOXO3, and a network of transcription factors were identified in the transitioning oocytes and somatic cells as potential upstream regulators that collectively may drive follicle activation. Taken together, we have developed a more precise characterization and selection method for studying staged-follicle cells, revealing several novel regulators of early folliculogenesis.

Tumorigenesis is associated with the deregulation of multiple processes, among which the glycosylation of lipids and proteins is one of the most extensively affected. However, in most cases, it remains unclear whether aberrant glycosylation is a cause, a link in the pathogenetic chain, or a mere consequence of tumorigenesis. In other cases, instead, studies have shown that aberrant glycans can promote oncogenesis. To comprehend how aberrant glycans are generated it is necessary to clarify the underlying mechanisms of glycan synthesis at the Golgi apparatus, which are still poorly understood. Important factors that determine the glycosylation potential of the Golgi apparatus are the levels and intra-Golgi localization of the glycosylation enzymes. These factors are regulated by the process of cisternal maturation which transports the cargoes through the Golgi apparatus while retaining the glycosylation enzymes in the organelle. This mechanism has till now been considered a single, house-keeping and constitutive function. Instead, we here propose that it is a mosaic of pathways, each controlling specific set of functionally related glycosylation enzymes. This changes the conception of cisternal maturation from a constitutive to a highly regulated function. In this new light, we discuss potential new groups oncogenes among the cisternal maturation machinery that can contribute to aberrant glycosylation observed in cancer cells. Further, we also discuss the prospects of novel anticancer treatments targeting the intra-Golgi trafficking process, particularly the cisternal maturation mechanism, to control/inhibit the production of pro-tumorigenic glycans.

Vascular tone is dependent on smooth muscle KATP channels comprising pore-forming Kir6.1 and regulatory SUR2B subunits, in which mutations cause Cantu syndrome. Unique among KATP isoforms, they lack spontaneous activity and require Mg-nucleotides for activation. Structural mechanisms underlying these properties are unknown. Here, we determined cryogenic electron microscopy structures of vascular KATP channels bound to inhibitory ATP and glibenclamide, which differ informatively from similarly determined pancreatic KATP channel isoform (Kir6.2/SUR1). Unlike SUR1, SUR2B subunits adopt distinct rotational "propeller" and "quatrefoil" geometries surrounding their Kir6.1 core. The glutamate/aspartate-rich linker connecting the two halves of the SUR-ABC core is observed in a quatrefoil-like conformation. Molecular dynamics simulations reveal MgADP-dependent dynamic tripartite interactions between this linker, SUR2B, and Kir6.1. The structures captured implicate a progression of intermediate states between MgADP-free inactivated, and MgADP-bound activated conformations wherein the glutamate/aspartate-rich linker participates as mobile autoinhibitory domain, suggesting a conformational pathway toward KATP channel activation.

Vitrification is a method for long-term biological sample cryopreservation without causing intra- and extra-cellular ice formation. We recently established a novel closed vitrification system to cryopreserve mouse ovarian follicles. Using the 3D alginate hydrogel encapsulated in vitro follicle growth (eIVFG) method, we demonstrated that compared to freshly-harvested follicles, vitrified follicles had normal follicle and oocyte reproductive outcomes. However, it is unknown whether vitrification preserves molecular signatures of folliculogenesis, which is the primary research focus in this study. Six fresh and six vitrified antral follicles grown from eIVFG were collected on day 8 for the whole single-follicle RNA sequencing. Principal component analysis (PCA) and Pearson's correlation analysis revealed that vitrified follicles had similar transcriptomic profiles to fresh follicles. There were 35 differentially expressed genes between vitrified and fresh follicles, however, none of those genes have been shown to be critical to folliculogenesis and oogenesis. Meanwhile, gene ontology (GO) and KEGG pathway analysis revealed that no GO terms or signaling pathways were significantly enriched. Furthermore, the expression of genes essential for the gonadotropin-dependent folliculogenesis and oogenesis were comparable between vitrified and fresh follicles. Taken together, these results demonstrate that vitrification preserves follicular cell transcriptome and molecular signatures of gonadotropin-dependent folliculogenesis in the eIVFG system, providing a robust model for fertility preservation, conservation of endangered species, and also establishing a high-content ovarian follicle biobank for studying ovarian biology and female reproductive toxicology.

The most promising probe to establish the Majorana or Dirac nature of the neutrino is the neutrinoless double beta decay and the effective neutrino mass would be evaluated by the knowledge of the corresponding nuclear matrix elements. Also measurements of the Double Charge Exchange (DCE) interactions of heavy ion beams can get information on them. The NUMEN experiment based on the pre-existing large acceptance MAGNEX spectrometer and integrated with new challenging components aims at measuring DCE cross sections using ion beams of unprecedented intensity (1013 pps) on specific isotopes at INFN-LNS in Catania. Expected rate on the sensitive area of about 0.15 m2 reaches up to about 5 Mpps, demanding for adequate detectors in measuring position, direction, energy, mass and charge of the ions produced by interactions. Gamma detectors surround a scattering chamber containing the target.

The identification of novel routes to synthetize functional molecules from bio-waste feedstock is one of the open challenges for a sustainable development of the chemical industry. However, most of the available feedstocks are complex mixtures of different molecules, which would ideally be processed without tedious and costly purification. The chemical complexity of such mixtures and their chemical transformations makes it difficult to build accurate mechanistic models for the optimization of the process conditions. To efficiently tackle this problem, we combine recent developments in machine learning (ML) and closed-loop optimization. The use of data-driven surrogate models coupled with design of experiments (DoE) algorithms have the potential to significantly speed up optimization of processes and suggest optimal conditions for synthetic routes, when no physical models are available.

The NUMEN project aims at accessing experimentally driven information on Nuclear Matrix Elements (NME) involved in the half-life of the neutrinoless double beta decay (0v beta beta). In this view measurements of Heavy Ion (HI) induced Double Charge Exchange (DCE) reaction cross sections are performed with high-accuracy. In particular, the (O-18,(18)Nc) and ((20)Nc, O-20) reactions are used as tools for beta(+)beta(+) and beta(-)beta(-) decays, respectively. In the experiments, performed at INFN - Laboratory Nazionali del Sud (LNS) in Catania, the beams are accelerated by the Superconducting Cyclotron (CS) and the reaction ejectiles are detected the MAGNEX magnetic spectrometer. The measured cross sections are challengingly low (a few nb), being the total reaction cross section much larger (a few b), thus a high sensitivity and a large rejection capability are demanded to the experimental set-up. This limits the present exploration to few selected isotopes of interest in the context of typically low-yield experimental runs. A major upgrade of the LNS facility is under way in order to increase the experimental dataset of more than two orders of magnitude, still preserving the high sensitivity of the present set-up. When accomplished this effort will make the INFN-LNS as the state-of-the-art research infrastructure for the systematic study of all the cases of interest for 0v beta beta. In this view, frontiers technologies are going to be adopted for the accelerator and the detection systems. In parallel, advanced theoretical models are being developed in order to extract the nuclear structure information from the measured cross sections

Significance Vascular KATP channels formed by the potassium channel Kir6.1 and its regulatory protein SUR2B maintain blood pressure in the physiological range. Overactivity of the channel due to genetic mutations in either Kir6.1 or SUR2B causes severe cardiovascular pathologies known as Cantú syndrome. The cryogenic electron microscopy structures of the vascular KATP channel reported here show multiple, dynamically related conformations of the regulatory subunit SUR2B. Molecular dynamics simulations reveal the negatively charged ED-domain in SUR2B, a stretch of 15 glutamate (E) and aspartate (D) residues not previously resolved, play a key MgADP-dependent role in mediating interactions at the interface between the SUR2B and Kir6.1 subunits. Our findings provide a mechanistic understanding of how channel activity is regulated by intracellular MgADP., Vascular tone is dependent on smooth muscle KATP channels comprising pore-forming Kir6.1 and regulatory SUR2B subunits, in which mutations cause Cantú syndrome. Unique among KATP isoforms, they lack spontaneous activity and require Mg-nucleotides for activation. Structural mechanisms underlying these properties are unknown. Here, we determined cryogenic electron microscopy structures of vascular KATP channels bound to inhibitory ATP and glibenclamide, which differ informatively from similarly determined pancreatic KATP channel isoform (Kir6.2/SUR1). Unlike SUR1, SUR2B subunits adopt distinct rotational “propeller” and “quatrefoil” geometries surrounding their Kir6.1 core. The glutamate/aspartate-rich linker connecting the two halves of the SUR-ABC core is observed in a quatrefoil-like conformation. Molecular dynamics simulations reveal MgADP-dependent dynamic tripartite interactions between this linker, SUR2B, and Kir6.1. The structures captured implicate a progression of intermediate states between MgADP-free inactivated, and MgADP-bound activated conformations wherein the glutamate/aspartate-rich linker participates as mobile autoinhibitory domain, suggesting a conformational pathway toward KATP channel activation.

PurposeThis paper aims to provide organizations with a new tool to make decisions related to a facility (building) selection process. Traditionally, value engineering (VE) applies the Value = Function/Cost formula to evaluate the worth of a product. In this paper, the VE-based facility-selection approach is proposed, where the cost of a facility is expressed in net present value (NPV) as it contains the net expense of purchasing or leasing a building as well as the time value of money. Also, a method of quantifying functions and involved risks of different facility choices is proposed.Design/methodology/approachThe framework of the VE-based facility-selection process is broken down into three steps: preparation, calculation/analysis and assessment. In the latter part of this paper, the authors share a sample analysis by illustrating the analysis and decision-making process when three hypothetical facility-selection options are available.FindingsThe sample analysis indicates that companies can get the lowest cost and risk while improving their functions to achieve the highest value by using the modified VE formula to drive an optimal option for company’s business expansion and facility-selection process.Originality/valueThis paper provides organizations with a strategic system and process to select proper facilities or buildings for business expansion. The VE approach suggested in this study can allow facility/real estate portfolio decision-makers to analyze financial and functional aspects of the facility at the same time and obtain the value coefficient when they choose a new facility from different options. Finally, they can select the best option, which has the highest value coefficient, given financial and functional considerations.

We investigate the relaxation dynamics of protein-polymer conjugates by neutron scattering spectroscopy to understand to which extent the coating of a protein by a polymer can replace water in promoting thermal structural fluctuations. For this purpose, we compare the dynamics of protein-polymer mixtures to that of conjugates with a variable number of polymers covalently attached to the protein. Results show that the flexibility of the protein is larger in protein-polymer mixtures than in native protein or in conjugates, even in the dry state. Upon hydration, both the native protein and the conjugate show equivalent dynamics, suggesting that the polymer grafted on the protein surface adsorbs all water molecules.

Vascular tone is dependent on smooth muscle KATP channels comprising pore-forming Kir6.1 and regulatory SUR2B subunits, in which mutations cause Cantú syndrome. Unique among KATP isoforms, they lack spontaneous activity and require Mg-nucleotides for activation. Structural mechanisms underlying these properties are unknown. Here, we determined the first cryoEM structures of vascular KATP channels bound to inhibitory ATP and glibenclamide, which differ informatively from similarly determined pancreatic KATP channel isoform (Kir6.2/SUR1). Unlike SUR1, SUR2B subunits adopt distinct rotational “propeller” and “quatrefoil” geometries surrounding their Kir6.1 core. The previously unseen ED-rich linker connecting the two halves of the SUR-ABC core is observed in a quatrefoil-like conformation. MD simulations reveal MgADP-dependent dynamic tripartite interactions between this linker, SUR2B and Kir6.1. The structures captured implicate a progression of intermediate states between MgADP-free inactivated and MgADP-bound activated conformations wherein the ED-rich linker participates as mobile autoinhibitory domain, suggesting a conformational pathway toward KATP channel activation.

The goal of NUMEN project is to access experimentally driven information on Nuclear Matrix Elements (NME) involved in the neutrinoless double beta decay (0νββ) by accurate measurements of the cross sections of heavy-ion induced double charge-exchange reactions. In particular, the (18O,18Ne) and (20Ne,20O) reactions are adopted as tools for β+β+and β−β−decays, respectively. The experiments are performed at INFN–Laboratory Nazionali del Sud (LNS) in Catania using the Superconducting Cyclotron to accelerate the beams and the MAGNEX magnetic spectrometer to detect the reaction products. The measured cross sections are very low, limiting the present exploration to few selected isotopes of interest in the context of typically low-yield experimental runs. In order to make feasible a systematic study of all the candidate nuclei, a major upgrade of the LNS facility is foreseen to increase the experimental yield by more than two orders of magnitude. To this purpose, frontier technologies are being developed for both the accelerator and the detection systems. An update description of the NUMEN project is presented here, focusing on recent achievements from the R&D activity.

We report a case of a newborn with two synchronous tumors—sialoblastoma and hepatoblastoma—diagnosed at 20 weeks of gestation by magnetic resonance imaging (MRI) and ultrasonography (US). The aim of this study was to describe the management of this case together with a review of the literature. Our patient had a large facial tumor associated with extremely high alpha-fetoprotein levels. Diagnosis of the tumors was made by surgical biopsy, showing typical features in both. Sialoblastoma is a potentially aggressive tumor. In our case, the Ki67 index in the sialoblastoma was between 20 and 30%, indicating a possibly unfavorable behavior. The infant underwent surgery and chemotherapy in different steps. Complete surgical resection with clean margins is considered to be the best treatment option for sialoblastoma. Only four similar cases were previously reported. Timely management by a multidisciplinary team is essential in these difficult cases. In our patient, outcome was good at the time of this report.

The search for neutrinoless double-beta (0νββ) decay is currently a key topic in physics, due to its possible wide implications for nuclear physics, particle physics, and cosmology. The NUMEN project aims to provide experimental information on the nuclear matrix elements (NMEs) that are involved in the expression of 0νββ decay half-life by measuring the cross section of nuclear double-charge exchange (DCE) reactions. NUMEN has already demonstrated the feasibility of measuring these tiny cross sections for some nuclei of interest for the 0νββ using the superconducting cyclotron (CS) and the MAGNEX spectrometer at the Laboratori Nazionali del Sud (LNS.) Catania, Italy. However, since the DCE cross sections are very small and need to be measured with high sensitivity, the systematic exploration of all nuclei of interest requires major upgrade of the facility. R&amp, D for technological tools has been completed. The realization of new radiation-tolerant detectors capable of sustaining high rates while preserving the requested resolution and sensitivity is underway, as well as the upgrade of the CS to deliver beams of higher intensity. Strategies to carry out DCE cross-section measurements with high-intensity beams were developed in order to achieve the challenging sensitivity requested to provide experimental constraints to 0νββ NMEs.

The Time Projection Chamber (TPC) is an ideal candidate to finely study the charged particle ionization in a gaseous medium. Large volume TPCs can be readout with a suitable number of channels offering a complete 3D reconstruction of a charged particle track, that is the sequence of its energy releases in the TPC gas volume. Moreover, He-based TPCs are very promising to study keV energy particles as nuclear recoils, opening the possibility for directional searches of Dark Matter (DM) and the study of Solar Neutrinos (SN). In this paper we report the analysis of the data acquired with a small TPC prototype (named LEMOn) built by the CYGNO collaboration that was exposed to a beam of 450 MeV electrons at the Beam Test Facility of National Laboratories of Frascati. LEMOn is operated with a He-C F 4 mixture at atmospheric pressure and is based on a Gas Electron Multipliers amplification stage that produces visible light collected by the high granularity and very good sensitivity of scientific CMOS camera. This type of readout – in conjunction with a fast light detection – allows a 3D reconstruction of the electrons tracks. The electrons are leaving a trail of clusters of ionizations corresponding to a few keV energy release each. Their study leads to predict a keV energy threshold and 1–10 mm longitudinal and 0.1–0.3 mm transverse position resolution (sigma) for nuclear recoils, very promising for the application of optically read out TPC to DM searches and SN measurements.