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γ-Aminobutyric acid type A receptors that incorporate α5 subunits (α5-GABAARs) are highly enriched in the hippocampus and are strongly implicated in control of learning and memory. Receptors located on pyramidal neuron dendrites have long been considered responsible, but here we report that mice in which α5-GABAARs have been eliminated from pyramidal neurons (α5-pyr-KO) continue to form strong spatial engrams and that they remain as sensitive as their pseudo-wild-type (p-WT) littermates to etomidate-induced suppression of place cells and spatial engrams. By contrast, mice with selective knockout in interneurons (α5-i-KO) no longer exhibit etomidate-induced suppression of place cells. In addition, the strength of spatial engrams is lower in α5-i-KO mice than p-WT littermates under control conditions. Consistent with the established role of the hippocampus in contextual fear conditioning, α5-i-KO mice resisted etomidate’s suppression of freezing to context, but so too did α5-pyr-KO mice, supporting a role for extra-hippocampal regions in the development of contextual fear memory. Overall, our results indicate that interneuronal α5-GABAARs serve a physiological role in promoting spatial learning and that they mediate suppression of hippocampus-dependent contextual memory by etomidate.

Point mutations in the β2 (N265S) and β3 (N265M) subunits of γ-amino butyric acid type A receptors (GABAARs) that render them insensitive to the general anesthetics etomidate and propofol have been used to link modulation of β2-GABAARs to sedation and β3-GABAARs to surgical immobility. These mutations also alter GABA sensitivity, and mice carrying the β3-N265M mutation have been reported to have impaired baseline memory. Here, we tested the effects of the β2-N265M and β3-N265M mutations on memory, movement, hotplate sensitivity, anxiety, etomidate-induced sedation, and intrinsic kinetics. We found that both β2-N265M and β3-N265M mice exhibited baseline deficits in the Context Preexposure Facilitation Effect learning paradigm. Exploratory activity was slightly greater in β2-N265M mice, but there were no changes in either genotype in anxiety or hotplate sensitivity. β2-N265M mice were highly resistant to etomidate-induced sedation, and heterozygous mice were partially resistant. In rapid solution exchange experiments, both mutations accelerated deactivation two- to three-fold compared to wild type receptors and prevented modulation by etomidate. This degree of change in the receptor deactivation rate is comparable to that produced by an amnestic dose of etomidate but in the opposite direction, indicating that intrinsic characteristics of GABAARs are optimally tuned under baseline conditions to support mnemonic function.

Objectives: Third wave cognitive behavioural therapies are increasingly used with children and adolescents. This meta-analysis aimed to determine the effectiveness of four third-wave interventions (acceptance and commitment therapy, compassion focused therapy, mindfulness-based cognitive therapy, and metacognitive therapy) for youth. Methods: Four electronic databases were used to identify randomized controlled trials, which tested effects related to health, well-being and functioning. Sensitivity analyses considering study quality were conducted and moderators were explored. Results: The results based on 50 RCTs meeting inclusion criteria indicated emotional symptoms/internalizing problems (g = −.68, 95% CI −.98 to −.37, k = 43, N = 3265), behavioural difficulties/externalizing problems (g = −.62, 95% CI −1.01 to −.22, k = 23, N = 1659), interference from difficulties (g = −.46, 95% CI −.87 to −.05, k = 21, N = 1786), third wave processes (g = .39, 95% CI .17 to .62, k = 22, N = 1900), wellbeing/flourishing (g = .76, 95% CI .35 to 1.17, k = 21, N = 1303) and physical health/pain (g = .72, 95% CI .01 to 1.44, k = 9, N = 1171) yielded significant effects. Effect for quality of life (g = .62, 95% CI −.08 to 1.31, k = 12, N = 1271) was non-significant. When analysing only those studies rated moderate-high quality, third wave interventions yielded significant superiority effects compared to controls for emotional symptoms/internalizing problems (g = −.55, 95% CI −.82 to −.27, k = 28, N = 2110), interference from difficulties (g = −.48, 95% CI −.90 to −.05, k = 21, N = 1605), third wave processes (g = .27, 95% CI .11 to .43, k = 18, N = 1692), well-being/flourishing (g = .50, 95% CI .18 to .81, k = 16, N = 1063), and quality of life (g = .32, 95% CI .04 to .60, k = 10, N = 1212). Behavioural difficulties/externalizing problems (g = −.38, 95% CI −.86 to .10, k = 15, N = 1351) and physical health/pain (g = .52, 95% CI −.14 to 1.17, k = 8, N = 1139) ceased to be significant. Widespread heterogeneity raised concerns about generalizability and follow-up data was relatively sparse. Conclusions: This meta-analysis finds promising results for use of third wave CBT with youth, though the review has limitations.

We present a chemical approach to profile fatty acid uptake in single cells. We use azide-modified analogues to probe the fatty acid influx and surface-immobilized dendrimers with dibenzocyclooctyne (DBCO) groups for detection. A competition between the fatty acid probes and BHQ2-azide quencher molecules generates fluorescence signals in a concentration-dependent manner. By integrating this method onto a microfluidics-based multiplex protein analysis platform, we resolved the relationships between fatty acid influx, oncogenic signaling activities, and cell proliferation in single glioblastoma cells. We found that p70S6K and 4EBP1 differentially correlated with fatty acid uptake. We validated that cotargeting p70S6K and fatty acid metabolism synergistically inhibited cell proliferation. Our work provided the first example of studying fatty acid metabolism in the context of protein signaling at single-cell resolution and generated new insights into cancer biology.

Top-down surface mixers are increasingly used in drinking water reservoirs to prevent the development of stratification, control cyanobacteria, and limit sediment release of soluble manganese. A targeted field investigation enabled the discrimination of artificial mixing by surface mixers from wind and convection in a shallow (6.6 m), eutrophic drinking water reservoir. Top-down surface mixers were effective at reducing vertical temperature and dissolved oxygen gradients over a 20 m radius, within which turbulent kinetic energy (TKE) input from the mixers exceeded the maximum TKE contribution from wind and convection. Meteorological conditions appeared to have a stronger influence beyond a 60 m radius from the mixers. Near-bed velocities measured using an Acoustic Doppler Velocimeter (ADV) ∼ 30 m north of the mixers were significantly lower when the mixers were not operating; when operating, ADV signal amplitude showed localized sediment resuspension. Cyanobacteria cell counts were high throughout the reservoir but counts of low-light adapted Planktothrix sp. were highest near the mixers, indicating mixer operation may improve growing conditions for Planktothrix. While the destratification goal of mixers was accomplished locally, the limited range of influence left >90% of the reservoir subject to diurnal stratification, anoxia, and potential internal loading of inorganic nutrients and soluble metals, restricting mixer effectiveness as an in-reservoir management technique to improve raw water quality in shallow systems.

Introduction: We evaluated potential factors influencing CGM uptake and sustained use to better understand the worsening disparities in CGM use among American youth with type 1 diabetes. Methods: This is a retrospective chart review of youth with type 1 diabetes less than 21 years old seen at a tertiary care center from 1/1/2021 to 11/5/2021. Most recent clinic note and prescriptions were used to determine CGM use (active, attrition, never) and whether CGM was accessed through pharmacy or durable medical equipment. Binary logistic regression was used to assess effects of age, sex, race/ethnicity, insulin regimen, health insurance, and CGM supplier on CGM uptake and sustained use. χ2 was used to assess the impact of language in Latinx youth. Results: Chart review identified 13youth (female 48.1%, NHW 43.7%, NHB 32.7%, Latinx 10.5%, publicly insured 52.1%) . Table 1 shows predictor variables associated with CGM use. NHW youth were more likely to have ever used CGM while CGM attrition was more common in NHB youth. Ever and active CGM use were less common in publicly insured youth and more common in pump users. CGM use was more common in Latinx English speakers than Spanish speakers (75.4% vs. 56.6%, p=0.03) . Conclusions: In an era of rapidly advancing technologies, efforts to support equity in diabetes technology use in publicly insured, minority, non-English speaking youth with type 1 diabetes may help to address disparities in health outcomes. Disclosure J. Grundman: Research Support; American Diabetes Association. M. Castro: None. A.E. Hoover: None. M. Monaghan: Employee; National Institute of Diabetes and Digestive and Kidney Diseases. Research Support; American Diabetes Association. R. Streisand: None. A.G. Perkins: None. J.K. Reilly: None. A.M. Richardson: None. B. Goggin: None. B.E. Marks: Advisory Panel; JDRF. Research Support; Dexcom, Inc., Tandem Diabetes Care, Inc. Funding American Diabetes Association (7-21-PDFHD-09)

We present here a cyclic peptide ligand, cy(WQETR), that binds to terbium ion (Tb(3+)) and enhances Tb(3+) luminescence intensity through the antenna effect. This peptide was identified through screening a cyclic peptide library against Tb(3+) with an apparent EC(50) of 540 μM. The tryptophan residue from the peptide directly interacts with the Tb(3+) ion, which provides access to a low-lying triplet excited state of the tryptophan. Direct excitation of this triplet state enables energy transfer to the Tb(3+) ion and enhances Tb(3+) luminescence intensity by 150 fold. We further showcase the application of this cy(WQETR)-Tb(3+) system by demonstrating the detection of tromethamine with a detection limit of 0.5 mM.

γ-aminobutyric acid type A receptors that incorporate α5 subunits (α5-GABAARs) are highly enriched in the hippocampus and are strongly implicated in control of learning and memory. Receptors located on pyramidal neuron dendrites have long been considered responsible, but here we report that their selective knockout from either interneurons (α5-i-KO) or pyramidal neurons (α5-pyr-KO) interferes with the ability of the general anesthetic etomidate to suppress contextual conditioning. Using Ca2+imaging of CA1 pyramidal neuron activity in freely exploring mice to assess hippocampal function directly, we found that etomidate blocked the development of place cells and spatial engrams in wild type (WT) and α5-pyr-KO mice, but not in α5-i-KO mice. In addition, α5-i-KO mice developed weaker spatial engrams than WT mice under control conditions. These findings show that interneuronal α5-GABAARs serve a physiological role in promoting spatial learning, and that they mediate the suppression of hippocampus-dependent memory by etomidate.

Land surface models and Earth system models that include Arctic landscapes must capture the abrupt hydrological transitions that occur during the annual thaw and deepening of the active layer. In this work, stable water isotopes (δ²H and δ¹⁸O) are used to appraise hydrologically significant transitions during annual landscape thaw at the Barrow Environmental Observatory (Utqiaġvik, Alaska). These hydrologically significant periods are then linked to annual shifts in the landscape energy balance, deduced from meteorological data and described by the microclimatic periods: Winter, Pre‐Melt, Melt, Post‐Melt, Summer, and Freeze‐Up. The tight coupling of the microclimatic periods with the hydrological transitions supports the use of microclimatic periods as a means of linking polygonal surface water hydrology to meteorological datasets, which provides a mechanism for improving the representation of polygonal surface water hydrology in process‐based models. Rayleigh process reconstruction of the isotopic changes revealed that 19% of winter precipitation was lost to sublimation prior to melting and that 23% of surface water was lost to evaporation during the first 10 days post‐melt. This agrees with evaporation rates reported in a separate study using an eddy covariance flux tower located nearby. An additional 17% was lost to evaporation during the next 33 days. Stable water isotopes are also used to identify the dominant sources of surface water to various hydrogeomorphological features prevalent in polygonal terrain (a lake, a low centre polygon centre, troughs within the rims of low centre polygons, flat centre polygon troughs, a high centre polygon trough, and drainages). Hydrogeomorphologies that retained significant old water or acted as snow drifts are isotopically distinct during the Melt Period and therefore are easily distinguished. Biogeochemical changes related to the annual thaw are also reported and coupled to the hydrological transitions, which provides insight into the sources and sinks of these ions to and from the landscape.

Introduction: We evaluated magnetic resonance spectroscopy (MRS) in United States military personnel with persistent symptoms after mild traumatic brain injury (mTBI), comparing over time two groups randomized to receive hyperbaric oxygen or sham chamber sessions and a third group of normative controls. Methods: Active-duty or veteran military personnel and normative controls underwent MRS outcome measures at baseline, 13 weeks (mTBI group only), and six months. Participants received 3.0 Tesla brain MRS for analysis of water-suppressed two-dimensional (2D) multivoxel 1H-MRS of the brain using point resolved spectroscopy (PRESS) with volume selection localized above the lateral ventricles and within the brain parenchyma, of which one voxel was chosen in each hemisphere without artifact. Script-based automatic data processing was used to assess N-acetylaspartate (NAA), creatine (Cr), and choline (Cho). Metabolite ratios for white matter were then calculated for NAA/Cr (Area), Cho/Cr (Area), and Cho/NAA (Area). These ratios were compared using standard analysis methodology. Results: There were no observable differences between participants with mTBI and normative controls nor any observable changes over time in the NAA/Cr (area), Cho/Cr (area), and Cho/NAA (area) ratios. Similarly, the control and injured participants were indistinguishable. Discussion: While participants with mild TBI showed no difference in MRS compared to normative controls, our results are limited by the few voxels chosen and potentially by less sensitive MRS markers.

Functional magnetic resonance imaging (fMRI) has been available commercially for clinical diagnostic use for many years. However, both clinical interpretation of fMRI by a neuroradiologist and quantitative analysis of fMRI data can require significant personnel resources that exceed reimbursement. In this report, a fully automated computerbased quantification methodology (Enumerated Auditory Response, EAR) has been developed to provide an auditory fMRI assessment of patients who have suffered a mild traumatic brain injury. Fifty-five study participants with interpretable auditory fMRI sequence data were assessed by EAR analysis, as well as both clinical radiologist fMRI interpretation and voxelwise general linear model (GLM) analysis. Comparison between the clinical interpretation and the two computer analysis methods resulted in 67% concordance (identical), 32% near-concordance (one level difference), and 1% discordant. Comparison between the clinical computer-based quantification (EAR) and GLM analysis yielded significant correlations in right and left ear responses (p

Increasing nitrogen (N) availability in Arctic soils could stimulate the growth of both plants and microorganisms by relieving the constraints of nutrient limitation. It was hypothesized that organic N addition to anoxic tundra soil would increase CH4 production by stimulating the fermentation of labile substrates, which is considered the rate-limiting step in anaerobic C mineralization. We tested this hypothesis through both field and lab-based experiments. In the field experiment, we injected a solution of 13C- and 15N-labeled glutamate 35 cm belowground at a site near Nome on the Seward Peninsula, Alaska, and observed the resulting changes in porewater geochemistry and dissolved greenhouse gas concentrations. The concentration of free glutamate declined rapidly within hours of injection, and the 15N label was recovered almost exclusively as dissolved organic N within 62 h. These results indicate rapid microbial assimilation of the added N and transformation into novel organic compounds. We observed increasing concentrations of dissolved CH4 and Fe(II), indicating rapid stimulation of methanogenesis and Fe(III) reduction. Low molecular weight organic acids such as acetate and propionate accumulated despite increasing consumption through anaerobic C mineralization. A laboratory soil column flow experiment using active layer soil collected from the same site further supported these findings. Glutamate recovery was low compared to a conservative bromide tracer, but concentrations of NO3− and NH4+ remained low, consistent with microbial uptake of the added N. Similar to the field experiment, we observed both increasing Fe(II) and organic acid concentrations. Together, these results support our hypothesis of increased fermentation in response to organic N addition and suggest that increasing N availability could accelerate CH4 production in tundra soils.

A common way to study the functional importance of N-methyl-D-aspartate receptors (NMDARs) in hippocampal memory-encoding circuits is by administering NMDAR antagonists. We recently compared the effects of (R,S)-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP), a competitive NMDAR antagonist, on suppression of memory in vivo versus suppression of NMDAR-mediated field EPSPs (fEPSPNMDA) and long-term potentiation (LTP) in vitro. Surprisingly, we found that concentrations that block contextual conditioning in vivo are ineffective at blocking the fEPSPNMDA or LTP in vitro. Here we tested one possible explanation for the mismatch – that the hippocampus is relatively resistant to CPP compared to other brain structures engaged in contextual fear conditioning. We used the context pre-exposure facilitation effect (CPFE) paradigm to isolate the hippocampal component of contextual learning, and in-vivo calcium imaging of place cells and spatial engrams to directly assess hippocampal spatial coding. We found that, by both measures, the active enantiomer (R)-CPP did interfere with hippocampal function at concentrations below those that block fEPSPs or LTP. We conclude that the alternative – that CPP interferes with memory by targeting NMDARs in interneurons rather than pyramidal neurons – is the more likely explanation.

Analyzing intracellular signalling protein activities in living cells promises a better understanding of the signalling cascade and related biological processes. We have previously developed cyclic peptide-based probes for analyzing intracellular AKT signalling activities, but these peptide probes were not cell-permeable. Implementing fusogenic liposomes as delivery vehicles could circumvent the problem when analyzing adherent cells, but it remained challenging to study suspension cells using similar approaches. Here, we present a method for delivering these imaging probes into suspension cells using digitonin, which could transiently perforate the cell membrane. Using U87, THP-1, and Jurkat cells as model systems representing suspended adherent cells, myeloid cells, and lymphoid cells, we demonstrated that low concentrations of digitonin enabled a sufficient amount of probes to enter the cytosol without affecting cell viability. We further combined this delivery method with a microwell single-cell chip and interrogated the AKT signalling dynamics in THP-1 and Jurkat cells, followed by immunofluorescence-based quantitation of AKT expression levels. We resolved the cellular heterogeneity in AKT signalling activities and showed that the kinetic patterns of AKT signalling and the AKT expression levels were related in THP-1 cells, but decoupled in Jurkat cells. We expect that our approach can be adapted to study other suspension cells.

In Arctic ecosystems, increasing temperatures are driving the expansion of nitrogen (N) fixing shrubs across tundra landscapes. The implications of this expansion to the biogeochemistry of Arctic ecosystems is of critical importance, yet many details about the form, location, and availability of N from these shrubs remain unknown. To address this knowledge gap, the spatiotemporal variability of nitrate (NO3−) and its environmental and edaphic controls were investigated at an alder (Alnus viridis spp. fruticosa) dominated permafrost tundra landscape in the Seward Peninsula, Alaska, USA. Soil pore water was collected from locations within alder shrubland growing along a well-drained hillslope and compared to soil pore water collected from locations outside (upslope, downslope, and between) the alder shrubland. δ15N and δ18O of soil pore water were consistent with the predicted range of NO3− produced through microbial degradation of N-rich alder shrub organic matter. Soil pore water collected within alder shrubland had an average NO3− concentration of (4.27 ± 8.02 mg L−1) and differed significantly from locations outside alder shrubland (0.23 ± 0.83 mg L−1; p 3− within and downslope of alder shrubland corresponded to precipitation events, where NO3− accumulated in the soil was flushed downslope during rainfall. Enrichment of both δ15N and δ18O isotopes at wetter downslope locations indicate that denitrification buffered the mobility and spatial extent of NO3−. These findings have important implications for nutrient production and mobility in N-limited permafrost systems that are experiencing shrub expansion in response to a warming Arctic.

Autoimmune diseases are characterized by a breakdown of immune tolerance partly due to environmental factors. The short-chain fatty acid acetate, derived mostly from gut microbial fermentation of dietary fiber, promotes antiinflammatory Tregs and protects mice from type 1 diabetes, colitis, and allergies. Here, we show that the effects of acetate extend to another important immune subset involved in tolerance, the IL-10–producing regulatory B cells (B10 cells). Acetate directly promoted B10 cell differentiation from mouse B1a cells both in vivo and in vitro. These effects were linked to metabolic changes through the increased production of acetyl-coenzyme A, which fueled the TCA cycle and promoted posttranslational lysine acetylation. Acetate also promoted B10 cells from human blood cells through similar mechanisms. Finally, we identified that dietary fiber supplementation in healthy individuals was associated with increased blood-derived B10 cells. Direct delivery of acetate or indirect delivery via diets or bacteria that produce acetate might be a promising approach to restore B10 cells in noncommunicable diseases.

Background Technological innovations have contributed to rapid changes in translational research, allowing greater amounts of shared data on an unprecedented scale. However, methods for involving patients in research have not kept pace with changes in research capacity. Modern tools offering more flexibility in the management of patient consent are needed. The CARPEM-LYNCH program aims to explore the acceptance and feasibility of the concept of dynamic informed consent. It is a pilot program to test a research platform at the interface between hospital follow-up clinical data and data provided by patients through the NutriNet-Santé e-cohort platform. Methods Patients diagnosed with Lynch Syndrome followed at the European hospital Georges Pompidou (HEGP) are recruited and followed-up within the NutriNet-Santé online platform. In addition to generic NutriNet questionnaires (very detailed data on diet, physical activity, lifestyle, etc.), participants receive specific questionnaires related to their syndrome, perception of nutritional risk factors, and quality of life. Clinical data collected during standard hospital care will be linked to NutriNet data for participants who provide a dynamic consent. This dispositive will allow to investigate multiple dimensions of dietary behaviors and their health impact in these at-risk patients. Results The pilot phase has started. The first 20 patients have been included, showing good acceptance of the dynamic consent. Qualitative analysis of their responses is ongoing to optimize tools before large-scale deployment and extension to other hospital centers. Conclusions This experience of merging hospital and e-cohort data through dynamic consent to advance knowledge on health impact of diet and lifestyle in Lynch patients opens up a multitude of perspectives. Key messages Dynamic informed consent offers opportunities for data sharing between clinicians, researchers and patients with a promising impact on translational research. Dynamic informed consent can provide practical and sustainable solutions to the challenges of recruiting and retaining participants, managing consent and it can also be a source of economic efficiency.

We report on a cyclic peptide that inhibits matrix metalloproteinase-2 (MMP2) activation with a low-nM-level potency. This inhibitor specifically binds to the D(570)-A(583) epitope on proMMP2 and interferes with the protein–protein interaction (PPI) between proMMP2 and tissue inhibitor of metalloproteinases-2 (TIMP2), thereby preventing the TIMP2-assisted proMMP2 activation process. We developed this cyclic peptide inhibitor through an epitope-targeted library screening process and validated its binding to proMMP2. Using a human melanoma cell line, we demonstrated the cyclic peptide’s ability to modulate cellular MMP2 activities and inhibit cell migration. These results provide the first successful example of targeting the PPI between proMMP2 and TIMP2, confirming the feasibility of an MMP2 inhibition strategy that has been sought after for 2 decades.

Whole blood is the preferred product for resuscitation of severe traumatic hemorrhage. It contains all the elements of blood that are necessary for oxygen delivery and hemostasis, in nearly physiologic ratios and concentrations. Group O whole blood that contains low titers of anti-A and anti-B antibodies (low titer group O whole blood) can be safely transfused as a universal blood product to patients of unknown blood group, facilitating rapid treatment of exsanguinating patients. Whole blood can be stored under refrigeration for up to 35 days, during which it retains acceptable hemostatic function, though supplementation with specific blood components, coagulation factors or other adjuncts may be necessary in some patients. Fresh whole blood can be collected from pre-screened donors in a walking blood bank to provide effective resuscitation when fully tested stored whole blood or blood components are unavailable and the need for transfusion is urgent. Available clinical data suggest that whole blood is at least equivalent if not superior to component therapy in the resuscitation of life-threatening hemorrhage. Low titer group O whole blood can be considered the standard of care in resuscitation of major hemorrhage.

Carbon dioxide (CO2) can be a significant resource input affecting the cost of algae biomass production on an industrial scale. Improvements in biofuel productivity therefore require characterization of CO2 use efficiency (CUE). RuBisCO saturation with CO2 is an important factor influencing biomass productivity. During CO2 fixation by RuBisCO, fractionation of carbon isotopes occurs, with preferential fixation of 12CO2, resulting in assimilation of the lighter isotope in algae. This photosynthetic discrimination (ΔDIC-algae), approximated by the difference between the δ13C of external medium and that of algae, is a function of the proportion of CO2 fixed relative to supply. ΔDIC-algae has been applied to the study of photosynthesis in algae over the past few decades and we have adopted the tool to characterize improvements in conditions optimized for biofuel productivity, such as controlled photobioreactors. We report the use of ΔDIC-algae as a tool for characterizing CO2 dynamics and RuBisCO saturation in Nannochloropsis salina CCMP 1776, a benchmark strain in algal biofuels research. We use ΔDIC-algae to describe the conditions under which RuBisCO saturation can be achieved to maximize biomass productivity. Our results suggest that determination of ΔDIC-algae can provide important feedback to support engineering and cultural improvements that can impact carbon use efficiency and biomass productivity.

Introduction et but de l’etude Les innovations technologiques ont contribue a l’evolution rapide de la recherche translationnelle, permettant le partage de plus grandes quantites de donnees. Toutefois, les methodes permettant d’impliquer les patients dans la recherche n’ont pas suivi l’evolution des capacites de recherche. Des outils modernes offrant une plus grande souplesse dans la gestion du consentement des patients sont necessaires. Le programme CARPEM-LYNCH vise a explorer l’acceptation et la faisabilite du concept de consentement dynamique eclaire en testant une plateforme de recherche situee a l’interface entre les donnees cliniques de suivi hospitalier et les donnees fournies par les patients dans le cadre de l’etude NutriNet-Sante. Materiel et methodes Les patients diagnostiques pour un syndrome de Lynch et suivis a l’hopital europeen Georges-Pompidou (HEGP) sont recrutes et suivis via la plateforme en ligne de l’etude NutriNet-Sante. Outre les questionnaires specifiques a NutriNet-Sante (donnees tres detaillees sur l’alimentation, l’activite physique, le mode de vie, etc.), les participants recoivent egalement des questionnaires en lien avec leur syndrome de Lynch (diagnostic et antecedents familiaux, sources d’informations nutritionnelles, exclusions alimentaires, qualite de vie). Les donnees cliniques collectees dans le cadre de leur suivi medical seront couplees aux donnees recueillies a partir de NutriNet-Sante, pour les participants qui donneront leur consentement. Ce dispositif permettra d’etudier les comportements alimentaires et leur impact sur la sante chez ces patients a risque de cancer. Resultats et analyse statistique La phase pilote de l’etude est lancee et inclut deja 50 premiers patients, ce qui prouve une bonne acceptation du consentement dynamique par les participants. Une analyse qualitative de leurs reponses est en cours afin d’optimiser la plateforme d’inclusion avant un deploiement a grande echelle du recrutement et son extension a d’autres centres hospitaliers. Conclusion Cette experience de couplage des donnees hospitalieres avec les donnees de la cohorte NutriNet-Sante sur la base du consentement des patients, contribue a faire progresser les connaissances sur l’impact de l’alimentation et du mode de vie sur la sante des patients porteurs du syndrome de Lynch. Elle offre par ailleurs, grâce au consentement dynamique eclaire, des possibilites de partage de donnees entre cliniciens, chercheurs et patients avec un impact prometteur sur la recherche translationnelle.

Breast tumors from young women under the age of 40 account for approximately 7% of cases and have a poor prognosis independent of established prognostic factors. We evaluated the patient population served by the Military Health System, where a disproportionate number of breast cancer cases in young women are seen and treated in a single universal coverage healthcare system.The Military Health System Repository and the DoD Central Registration databases were used to identify female breast cancer patients diagnosed or treated at military treatment facilities from 1998 to 2007.10,066 women were diagnosed with invasive breast cancer at DoD facilities from 1998 to 2007, of which 11.3% (1139), 23.4% (2355) and 65.2% (6572) were 40, 40-49 and 50 years old (yo), respectively, at diagnosis. 53% in the 40 yo cohort were white, 25% were African American (AA) and 8% were Hispanic, with 14% undisclosed. Breast cancer in women diagnosed 40 yo was more high grade (p 0.0001), Stage II (p 0.0001) and ER negative (p 0.0001). There was a higher rate of bilateral mastectomies among the women 40 compared to those 40-49 and 50 (18.4% vs. 9.1% and 5.0%, respectively). Independent of disease stage, chemotherapy was given more frequently to 40 yo (90.43%) and 40-49 yo (81.44%) than ≥ 50 yo (53.71%). The 10-year overall survival of younger women was similar to the ≥ 50 yo cohort. Outcomes in the African American and Hispanic subpopulations were comparable to the overall cohort.Younger women had a similar overall survival rate to older women despite receiving more aggressive treatment.

Purpose To review and analyze the key structural considerations secondaries funds and funds of funds must consider when negotiating credit facilities secured by limited partnership (LP) interests. Design/methodology/approach This article provides an overview of the primary issues that arise with credit facilities secured by LP interests. These issues include the ability to provide a perfected security interest in LP interests, understanding a credit facility’s borrowing base and advance rates, and the potential impact of certain types of events of default. Findings Secondaries funds and funds of funds have raised significant amounts of equity capital in recent years. These funds acquire portfolios of LP interests and are increasingly deploying leverage to amplify the returns of these portfolios and provide these funds with a limited degree of liquidity. The leverage is secured by the LP interests. The credit facilities that the funds are structuring and negotiating present a host of issues unique to this type of fund finance. Provided the facilities are properly structured and negotiated, secondaries funds and funds-of-funds borrowers will be able to use these facilities to help meet investment-return objectives and address important portfolio-management needs. Practical implications Secondaries funds and funds of funds can benefit from leverage secured by their portfolios of LP interests. This article provides a road map for borrowers when structuring and negotiating these credit facilities. Originality/value Practical analysis from a premier corporate law firm on the issues presented by the increasing use of credit facilities by secondaries funds and funds of funds.

Scale-up of microalgal biotechnology to provide large quantities of biofuel, lipids, and coproducts is not fully developed because of the large needs for nutrients, water, land, solar insolation, and CO2/carbon supplies. Wastewaters, including oil and gas produced water (PW), may supply a portion of these needs in regions with insufficient fresh water resources. PW is a challenging water resource for this use because of variable salinity, geochemical complexity, and the presence of biologically toxic components. In this paper we review PW volumes, quality, and use in media for microalgae production in the southwestern US, Australia, and Oman. We also include data from the southwestern US, referencing previously unpublished results from the National Alliance for Biofuels and Bioproducts (NAABB) consortium research project. We include a Supplementary Information section that explores cultivation of multiple microalgae species in PW and examines the carbon utilization process, all work performed in support of the NAABB field program. Strains of algae tested in the reviewed papers include Nannochloropsis, Dunalliella, Scenedesmus, and several mixed or unknown cultures. We conclude that the use of PW in algae cultivation is feasible, if the additional complexity of the water resource is accounted for in developing media formulations and in understanding metals uptake by the algae. We recommend additional work to standardize growth testing in PW, better and more thorough chemical analysis, and geochemical modeling of the PW used in media. Expanded strain testing in PW media will identify improved strains tolerant of PW in algae cultivation.

Functional magnetic resonance imaging (fMRI) has been available commercially for clinical diagnostic use for many years. However, both clinical interpretation of fMRI by a neuroradiologist and quantitative analysis of fMRI data can require significant personnel resources that exceed reimbursement. In this report, a fully automated computer-based quantification methodology (Enumerated Auditory Response, EAR) has been developed to provide an auditory fMRI assessment of patients who have suffered a mild traumatic brain injury. Fifty-five study participants with interpretable auditory fMRI sequence data were assessed by EAR analysis, as well as both clinical radiologist fMRI interpretation and voxelwise general linear model (GLM) analysis. Comparison between the clinical interpretation and the two computer analysis methods resulted in 67% concordance (identical), 32% nearconcordance (one level difference), and 1% discordant. Comparison between the clinical computer-based quantification (EAR) and GLM analysis yielded significant correlations in right and left ear responses (p⟨0.05) for the full subject group. Automated fMRI quantification analysis equivalent to EAR might be appropriate for both future research projects with constrained resources, as well as possible routine clinical use.

Objective To assess both the feasibility and value of conducting an external quality assurance programme concerning technical aspects of cytopathology laboratory practice, and the interest by laboratories in enrolling in such a programme. Methods Six technical surveys, comprising staining exercises and questionnaires relating to laboratory practice, were distributed over a 4-year period to the approximately 220 laboratories enrolled in the RCPAQAP Cytopathology slide survey modules. Staining exercises using the Papanicolaou and Romanowsky techniques, the preparation of urine and body fluid specimens and immunocytochemistry on the cell block material were assessed. Accompanying relevant questionnaires were included, and one survey comprised a questionnaire alone concerning the collection of urinary tract and body fluid samples. Results Provision of an external cytopathology technical module was feasible for the RCPAQAP and participation rates (maximum of 87% per survey; average 68% for stained slides and 66% for questionnaires) were commendable, particularly considering these were optional undertakings with some exercises not applicable to all laboratories. The great majority of submitted slides were scored as satisfactory, and there was an especially high standard for the immunocytochemical staining exercise with 95% considered satisfactory, including 50.6% with a perfect score. Reasons for suboptimal scores were provided for potential quality improvement for interested laboratories. A wealth of information relating to laboratory practice was provided to the RCPAQAP which was collated and summarised for laboratory use. Conclusions The provision of a technical module in cytopathology is both a feasible and valuable undertaking of interest to laboratories which should become standard practice for cytopathology external quality assurance providers.

Resume La medecine de precision prend une place grandissante dans la prise en charge therapeutique des patients en oncologie. Le developpement des techniques de sequencage a haut debit ou de nouvelle generation permet l’analyse d’un plus grand nombre de genes a partir d’un meme echantillon. Dans cet article, nous developpons l’impact de ces nouvelles analyses dans la prise en charge des patients, avec une limite qui devient flou entre l’analyse somatique et constitutionnelle, et le questionnement ethique que cela souleve.

We present here a novel chemical method to continuously analyze intracellular AKT signaling activities at single-cell resolution, without genetic manipulations. A pair of cyclic peptide-based fluorescent probes were developed to recognize the phosphorylated Ser474 site and a distal epitope on AKT. A Förster resonance energy transfer signal is generated upon concurrent binding of the two probes onto the same AKT protein, which is contingent upon the Ser474 phosphorylation. Intracellular delivery of the probes enabled dynamic measurements of the AKT signaling activities. We further implemented this detection strategy on a microwell single-cell platform, and interrogated the AKT signaling dynamics in a human glioblastoma cell line. We resolved unique features of the single-cell signaling dynamics following different perturbations. Our study provided the first example of monitoring the temporal evolution of cellular signaling heterogeneities and unveiled biological information that was inaccessible to other methods.

Arctic soils contain a large pool of terrestrial C and are of interest due to their potential for releasing significant carbon dioxide (CO2) and methane (CH4) to the atmosphere. Due to substantial landscape heterogeneity, predicting ecosystem-scale CH4 and CO2 production is challenging. This study assessed dissolved inorganic carbon (DIC = Σ (total) dissolved CO2) and CH4 in watershed drainages in Barrow, Alaska as critical convergent zones of regional geochemistry, substrates, and nutrients. In July and September of 2013, surface waters and saturated subsurface pore waters were collected from 17 drainages. Based on simultaneous DIC and CH4 cycling, we synthesized isotopic and geochemical methods to develop a subsurface CH4 and DIC balance by estimating mechanisms of CH4 and DIC production and transport pathways and oxidation of subsurface CH4. We observed a shift from acetoclastic (July) toward hydrogenotropic (September) methanogenesis at sites located toward the end of major freshwater drainages, adjacent to salty estuarine waters, suggesting an interesting landscape-scale effect on CH4 production mechanism. The majority of subsurface CH4 was transported upward by plant-mediated transport and ebullition, predominantly bypassing the potential for CH4 oxidation. Thus, surprisingly, CH4 oxidation only consumed approximately 2.51 ± 0.82% (July) and 0.79 ± 0.79% (September) of CH4 produced at the frost table, contributing to

The nitrate (NO3−) dual isotope approach was applied to snowmelt, tundra active layer pore waters, and underlying permafrost in Barrow, Alaska, USA, to distinguish between NO3− derived from atmospheric deposition versus that derived from microbial nitrification. Snowmelt had an atmospheric NO3− signal with δ15N averaging −4.8 ± 1.0‰ (standard error of the mean) and δ18O averaging 70.2 ± 1.7‰. In active layer pore waters, NO3− primarily occurred at concentrations suitable for isotopic analysis in the relatively dry and oxic centers of high-centered polygons. The average δ15N and δ18O of NO3− from high-centered polygons were 0.5 ± 1.1‰ and −4.1 ± 0.6‰, respectively. When compared to the δ15N of reduced nitrogen (N) sources, and the δ18O of soil pore waters, it was evident that NO3− in high-centered polygons was primarily from microbial nitrification. Permafrost NO3− had δ15N ranging from approximately −6‰ to 10‰, similar to atmospheric and microbial NO3−, and highly variable δ18O ranging from approximately −2‰ to 38‰. Permafrost ice wedges contained a significant atmospheric component of NO3−, while permafrost textural ice contained a greater proportion of microbially derived NO3−. Large-scale permafrost thaw in this environment would release NO3− with a δ18O signature intermediate to that of atmospheric and microbial NO3. Consequently, while atmospheric and microbial sources can be readily distinguished by the NO3− dual isotope technique in tundra environments, attribution of NO3− from thawing permafrost will not be straightforward. The NO3− isotopic signature, however, appears useful in identifying NO3− sources in extant permafrost ice.

Mixing models are powerful tools for identifying biogeochemical sources and determining mixing fractions in a sample. However, identification of actual source contributors is often not simple, and source compositions typically vary or even overlap, significantly increasing model uncertainty in calculated mixing fractions. This study compares three probabilistic methods, Stable Isotope Analysis in R (SIAR), a pure Monte Carlo technique (PMC), and Stable Isotope Reference Source (SIRS) mixing model, a new technique that estimates mixing in systems with more than three sources and/or uncertain source compositions. In this paper, we use nitrate stable isotope examples (δ15N and δ18O) but all methods tested are applicable to other tracers. In Phase I of a three-phase blind test, we compared methods for a set of six-source nitrate problems. PMC was unable to find solutions for two of the target water samples. The Bayesian method, SIAR, experienced anchoring problems, and SIRS calculated mixing fractions that most closely approximated the known mixing fractions. For that reason, SIRS was the only approach used in the next phase of testing. In Phase II, the problem was broadened where any subset of the six sources could be a possible solution to the mixing problem. Results showed a high rate of Type I errors where solutions included sources that were not contributing to the sample. In Phase III some sources were eliminated based on assumed site knowledge and assumed nitrate concentrations, substantially reduced mixing fraction uncertainties and lowered the Type I error rate. These results demonstrate that valuable insights into stable isotope mixing problems result from probabilistic mixing model approaches like SIRS. The results also emphasize the importance of identifying a minimal set of potential sources and quantifying uncertainties in source isotopic composition as well as demonstrating the value of additional information in reducing the uncertainty in calculated mixing fractions.

A sensitive chemical aerosol Raman detector (CARD) has been developed for the trace detection and identification of chemical particles in the ambient atmosphere. CARD includes an improved aerosol concentrator with a concentration factor of about 40 and a CCD camera for improved detection sensitivity. Aerosolized isovanillin, which is relatively safe, has been used to characterize the performance of the CARD. The limit of detection (SNR = 10) for isovanillin in 15 s has been determined to be 1.6 pg/cm3, which corresponds to 6.3 × 109 molecules/cm3 or 0.26 ppb. While less sensitive, CARD can also detect gases. This paper provides a more detailed description of the CARD hardware and detection algorithm than has previously been published.

Mixtures of water and inorganic acids show differences in isotopic composition between the vapor and liquid phases, reportedly even at an azeotrope. A recent reactive azeotropy analysis rationalized this by allowing for differences in stable isotope composition between bound solvation complexes and the bulk liquid. The objective of this study is to determine whether or not significant differences in stable isotope composition exist between bound solvation complexes involving inorganic acids and the bulk solution. Rayleigh distillation of a solution of nitric acid and water was conducted. During the distillation, it was expected that bulk water would be vaporized first while (bound) water involved with hydrating the acid would be vaporized at later times along with the acid. Samples of both distillate and residual liquid were collected over time intervals and analyzed to determine oxygen and nitrogen stable isotope composition using isotope ratio mass spectrometry. As a reference, comparative samples were collected from Rayleigh distillation of water alone. The measured oxygen fractionation factors suggest that the acid is preferentially hydrated with 18O-enriched water, compared to the unbound, or bulk, water. Preferential coordination of 18O-enriched water with the dissolved acid is consistent with heavier water forming stronger bonds relative to light water. The results, although obtained under non-equilibrum conditions, suggest that isotope-dependent partitioning between bulk solvent and dissolved complexes may prove necessary in developing accurate descriptions of isotope fractionation during vapor-liquid equilibrium.

Improved predictions of drought require an understanding of natural and human-induced climate variability. Long-term records across glacial–interglacial cycles provide the natural component of variability, however few such records exist for the southwestern United States (US) and quantitative or semi-quantitative records of precipitation are absent. Here we use the hydrogen isotope (δD) value of C28 n-alkanoic acid in lacustrine sediments of Pleistocene age to reconstruct δD values of precipitation in northern New Mexico over two glacial–interglacial cycles (∼550,000–360,000 years before present) and obtain a record of monsoon strength. Overall, reconstructed δD values range from −53.8‰ to −94.4‰, with a mean value of −77.5 ± 8‰. Remarkably, this variation falls within the measured present-day summer monsoonal and winter weighted means (−50.3 ± 3‰ and −106.4 ± 20‰ respectively), suggesting that processes similar to those of present time also controlled precipitation during Marine Isotope Stage (MIS) 13 to 10. Using the δD summer monsoonal and winter mean values as end-members, we interpret our reconstructed δD record of precipitation as a direct, and semi-quantitative, indicator of monsoon strength during MIS 13 to 10. Interglacial periods were characterized by greater monsoon strength but also greater variability compared to glacial periods. Pronounced cycles in the strength of the monsoon occurred during interglacial periods and in general were positively correlated with maximum mean annual temperatures. Our estimates of monsoon strength are supported by independent proxies of ecosystem productivity, namely, TOC, δ13C of TOC and Si/Ti ratio and warm pollen taxa Juniperus and Quercus. Interglacial variability in the strength of the monsoon resembles a response to the land-sea surface temperature contrast (LSTC) except for the early part of MIS 11. During this period, LSTC would have remained relatively strong while monsoonal strength decreased to a minimum. This minimum occurred following the warmest interval of MIS 11, suggesting a more complex driving of monsoon strength during warm periods. In addition, this period of monsoon minimum coincided with a core section of mud-cracked sediments that suggest low monsoonal precipitation was an important factor in the onset of drought. Our estimates of monsoon strength represent a record of natural variability in the region that is relevant to present time, in particular the variability during interglacial MIS 11, which is considered an analog for the current interglacial. Our results suggest that natural variability can cause significant reductions in monsoonal precipitation with the implication of a potentially adverse effect from sustained warming.

During a carbon sequestration field study to simulate the impact of CO 2 migration on shallow groundwater chemistry, the isotope composition of dissolved inorganic carbon ( δ 13 C DIC ) and dissolved strontium ( 87 Sr/ 86 Sr) were evaluated as tracers. Dissolved CO 2 in groundwater was introduced using a closed-loop dipole-style well field situated in a shallow sand-dominated aquifer. Baseline δ 13 C DIC values, oxygen and hydrogen isotope ratios, and 87 Sr/ 86 Sr values of groundwater were established in four monitoring wells (MW-1 to 4) and one up-gradient background well (BG-1) prior to the introduction of dissolved CO 2 . Baseline groundwater δ 13 C DIC-PDB , oxygen ( δ 18 O SMOW ) and hydrogen ( δ D SMOW ) stable isotope values averaged −17, −4.1 and −19.5‰, respectively. Groundwater 87 Sr/ 86 Sr baseline values averaged 0.70840 at MW-3 and 0.70818 at MW-2. Arrival of the dissolved CO 2 plume at the monitoring wells is modeled using a 1-D analytical equation, which yields breakthrough curves with flow velocities that are consistent with prior numerical modeling estimates. The δ 13 C DIC-PDB rose to an average steady-state value of 0.16 ± 0.3‰ during the test; δ 18 O and δ D of water did not change from their baseline values. 87 Sr/ 86 Sr dropped sharply by 0.00022 at MW-3 and 0.00005 at MW-2 in the first two weeks after plume arrival at the wells, and then slowly increased toward baseline values, correlating with the behavior of dissolved Na, K, Ca, Sr and Si. Carbonate dissolution and desorption from organic matter and Fe-bearing phases at the low-pH plume front is the likely mechanism producing this behavior. The δ 13 C DIC and the 87 Sr/ 86 Sr of dissolved strontium served as excellent tracers of plume movement during this experiment.

There is currently a lack of suitable objective endpoints to measure disease progression in Duchenne muscular dystrophy (DMD). Emerging research suggests that diffusion tensor imaging (DTI) has potential as an outcome measure for the evaluation of skeletal muscle injury.The objective of this study was to evaluate the potential of DTI as quantitative magnetic resonance imaging (MRI) markers of disease severity in DMD.Thirteen consecutive boys (8.9 years ± 3.0 years) with DMD were evaluated using DTI. Fractional anisotropy (FA) and apparent diffusion coefficient (ADC) were compared with clinical outcome measures of manual muscle testing and MRI determinations of muscle fat fraction (MFF) in the right lower extremity.Both MRI measures of FA and ADC strongly correlated with age and muscle strength. Values for FA positively correlated with age and negatively correlated with muscle strength (r = 0.78 and -0.96; both P ≤ 0.002) while measures of ADC negatively correlated age, but positively correlated with muscle strength (r = -0.87 and 0.83; both P ≤ 0.0004). Additionally, ADC and FA strongly correlated with MFF (r = -0.891 and 0.894, respectively; both P ≤ 0.0001). Mean MMF was negatively correlated with muscle strength (r = -0.89, P = 0.0001).DTI measures of muscle structure strongly correlated with muscle strength and adiposity in boys with DMD in this pilot study, although these markers may be more reflective of fat replacement rather than muscle damage in later stages of the disease. Further studies in presymptomatic younger children are needed to assess the ability of DTI to detect early changes in DMD.

Over the past decade, breakthroughs in the generation and control of ultrafast high-field terahertz (THz) radiation have led to new spectroscopic methodologies for the study of light-matter interactions in the strong-field limit. In this review, we will outline recent experimental demonstrations of non-linear THz material responses in materials ranging from molecular gases, to liquids, to varieties of solids – including semiconductors, nanocarbon, and correlated electron materials. New insights into how strong THz fields interact with matter will be discussed in which a THz field can act as either a non-resonant electric field or a broad bandwidth pulse driving specific resonances within it. As an emerging field, non-linear THz spectroscopy shows promise for elucidating dynamic problems associated with next generation electronics and optoelectronics, as well as for demonstrating control over collective material degrees of freedom.