Covalently linking oligomerization-impaired GlpF protomers does not completely re-establish wild-type channel activity

Integral membrane proteins of the aquaporin family facilitate rapid water flux across cellular membranes in all domains of life. although the water-conducting pore is clearly defined in an aquaporin monomer, all aquaporins assemble into stable tetramers. in order to investigate the role of protomer–protomer interactions, we analyzed the activity of heterotetramers containing increasing fractions of mutated monomers, which have an impaired oligomerization propensity and activity. in order to enforce interaction between the protomers, we designed and analyzed a genetically fused homotetramer of glpf, the aquaglyceroporin of the bacterium escherichia coli (e. coli). however, increasing fractions of the oligomerization-impaired mutant glpf e43a affected the activity of the glpf heterotetramer in a nearly linear manner, indicating that the reduced protein activity, caused by the introduced mutations, cannot be fully compensated by simply covalently linking the monomers. taken together, the results underline the importance of exactly positioned monomer–monomer contacts in an assembled glpf tetramer

The think/no-think alcohol task: a new paradigm for assessing memory suppression in alcohol-related contexts

Background Research with the Think/No-Think (TNT) task has shown that voluntary suppression of an unwanted memory may lead to its later forgetting. To date, however, no study has assessed the memory suppression abilities in alcohol-related contexts despite the potential implications that it might have for alcohol research. With this aim, we developed a new version of the TNT paradigm, the TNT Alcohol (TNTA) task, which consists of 36 neutral pictures paired with 36 alcohol/no-alcohol images that are instructed to be suppressed or recollected. Methods Electroencephalographic activity was recorded from 64 electrodes while 20 young healthy females performed the TNTA task. The event-related potentials (ERPs) typically involved in memory suppression/recollection were analyzed, namely the fronto-central N2, the late parietal positivity (LPP), and the frontal slow wave (FSW). Results Findings revealed reduced recall for previously learned images that were subsequently instructed to be suppressed (No-Think) relative to those instructed to be retrieved (Think) and those not cued to be suppressed or retrieved (Baseline). This reduction seemed to be more prominent for alcohol-related memories. In addition, ERP analysis showed that compared to attempts of recollection, attempts of memory suppression were associated with attenuated LPP amplitude-more pronounced for alcohol-related memories-(indicating reduced conscious recollection for No-Think images) as well as with increased FSW (suggesting strategic control aiming at decrease accessibility of unwanted memories). Conclusions These results replicate and extend previously reported behavioral and ERP findings in the TNT paradigm and suggest that the TNTA task may be a useful instrument to measure the ability to suppress alcohol-related memories.This study was supported by the project NORTE-01-0145-FEDER-028672, funded by the Portuguese Foundation for Science and Technology (FCT) and the European Regional Development Fund (FEDER). EL-C and AC were supported by a Postdoctoral Fellowship of the FCT (SFRH/BPD/109750/2015 and SFRH/BPD/91440/2012, respectively), as well as by the Psychology Research Centre (UID/PSI/01662/2013), co-financed by FEDER through COMPETE2020 under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007653)

Design and Expression of a Dimeric Form of the Human Immunodeficiency Virus Type 1 Antibody 2G12 with Increased Neutralization Potency

The antigen-binding fragment of the broadly neutralizing Human Immunodeficiency Virus Type 1 (HIV-1) antibody 2G12 has an unusual 3D domain-swapped structure with two aligned combining sites that facilitates recognition of its carbohydrate epitope on gp120. When expressed as an intact IgG, 2G12 formed typical IgG monomers containing two combining sites and a small fraction of a higher molecular weight species, which showed a significant increase in neutralization potency (50- to 80-fold compared to 2G12 monomer) across a range of clade A and B strains of HIV-1. Here we show that the higher molecular weight species corresponds to a 2G12 dimer containing four combining sites, and present a model for how intermolecular 3D domain swapping could create a 2G12 dimer. Based on the structural model for a 3D domain-swapped 2G12 dimer, we designed and tested a series of 2G12 mutants predicted to increase the ratio of 2G12 dimer to monomer. We report a mutation that effectively increases the 2G12 dimer/monomer ratio without decreasing the expression yield. Increasing the proportion of 2G12 dimer compared with monomer could lead to a more potent reagent for gene therapy or passive immunization

HIV Testing of At Risk Patients in a Large Integrated Health Care System

OBJECTIVE: Early identification of HIV infection is critical for patients to receive life-prolonging treatment and risk-reduction counseling. Understanding HIV screening practices and barriers to HIV testing is an important prelude to designing successful HIV screening programs. Our objective was to evaluate current practice patterns for identification of HIV. METHODS: We used a retrospective cohort analysis of 13,991 at-risk patients seen at 4 large Department of Veterans Affairs (VA) health-care systems. We also reviewed 1,100 medical records of tested patients. We assessed HIV testing rates among at-risk patients, the rationale for HIV testing, and predictors of HIV testing and of HIV infection. RESULTS: Of the 13,991 patients at risk for HIV, only 36% had been HIV-tested. The prevalence of HIV ranged from 1% to 20% among tested patients at the 4 sites. Approximately 90% of patients who were tested had a documented reason for testing. CONCLUSION: One-half to two-thirds of patients at risk for HIV had not been tested within our selected VA sites. Among tested patients, the rationale for HIV testing was well documented. Further testing of at-risk patients could clearly benefit patients who have unidentified HIV infection by providing earlier access to life-prolonging therapy

Constraining long-term denudation and faulting history in intraplate regions by multisystem thermochronology: An example of the Sudetic Marginal Fault (Bohemian Massif, central Europe)

The Rychlebské hory Mountain region in the Sudetes (NE Bohemian Massif) provides a natural laboratory for studies of postorogenic landscape evolution. This work reveals both the exhumation history of the region and the paleoactivity along the Sudetic Marginal Fault (SMF) using zircon (U-Th)/He (ZHe), apatite fission track (AFT), and apatite (U-Th)/He (AHe) dating of crystalline basement and postorogenic sedimentary samples. Most significantly, and in direct contradiction of traditional paleogeographic reconstructions, this work has found evidence of a large Cretaceous sea and regional burial (to >6.5 km) of the Carboniferous-Permian basement in the Late Cretaceous (~95–80 Ma). During the burial by sediments of the Bohemian Cretaceous Basin System, the SMF acted as a normal fault as documented by offset ZHe ages across the fault. At 85–70 Ma, the basin was inverted, Cretaceous strata eroded, and basement blocks were exhumed to the near surface at a rate of ~300 m/Ma as evidenced by Late Cretaceous–Paleocene AFT ages and thermal modeling results. There is no appreciable difference in AFT and AHe ages across the fault, suggesting that the SMF acted as a reverse fault during exhumation. In the late Eocene–Oligocene, the basement was locally heated to <70°C by magmatic activity related to opening of the Eger rift system. Neogene or younger thermal activity was not recorded in the thermochronological data, confirming that late Cenozoic uplift and erosion of the basement blocks was limited to less than ∼1.5 km in the study area

Clinical Sequencing Exploratory Research Consortium: Accelerating Evidence-Based Practice of Genomic Medicine

Despite rapid technical progress and demonstrable effectiveness for some types of diagnosis and therapy, much remains to be learned about clinical genome and exome sequencing (CGES) and its role within the practice of medicine. The Clinical Sequencing Exploratory Research (CSER) consortium includes 18 extramural research projects, one National Human Genome Research Institute (NHGRI) intramural project, and a coordinating center funded by the NHGRI and National Cancer Institute. The consortium is exploring analytic and clinical validity and utility, as well as the ethical, legal, and social implications of sequencing via multidisciplinary approaches; it has thus far recruited 5,577 participants across a spectrum of symptomatic and healthy children and adults by utilizing both germline and cancer sequencing. The CSER consortium is analyzing data and creating publically available procedures and tools related to participant preferences and consent, variant classification, disclosure and management of primary and secondary findings, health outcomes, and integration with electronic health records. Future research directions will refine measures of clinical utility of CGES in both germline and somatic testing, evaluate the use of CGES for screening in healthy individuals, explore the penetrance of pathogenic variants through extensive phenotyping, reduce discordances in public databases of genes and variants, examine social and ethnic disparities in the provision of genomics services, explore regulatory issues, and estimate the value and downstream costs of sequencing. The CSER consortium has established a shared community of research sites by using diverse approaches to pursue the evidence-based development of best practices in genomic medicine

Effects of a high-dose 24-h infusion of tranexamic acid on death and thromboembolic events in patients with acute gastrointestinal bleeding (HALT-IT): an international randomised, double-blind, placebo-controlled trial

Background: Tranexamic acid reduces surgical bleeding and reduces death due to bleeding in patients with trauma. Meta-analyses of small trials show that tranexamic acid might decrease deaths from gastrointestinal bleeding. We aimed to assess the effects of tranexamic acid in patients with gastrointestinal bleeding. Methods: We did an international, multicentre, randomised, placebo-controlled trial in 164 hospitals in 15 countries. Patients were enrolled if the responsible clinician was uncertain whether to use tranexamic acid, were aged above the minimum age considered an adult in their country (either aged 16 years and older or aged 18 years and older), and had significant (defined as at risk of bleeding to death) upper or lower gastrointestinal bleeding. Patients were randomly assigned by selection of a numbered treatment pack from a box containing eight packs that were identical apart from the pack number. Patients received either a loading dose of 1 g tranexamic acid, which was added to 100 mL infusion bag of 0·9% sodium chloride and infused by slow intravenous injection over 10 min, followed by a maintenance dose of 3 g tranexamic acid added to 1 L of any isotonic intravenous solution and infused at 125 mg/h for 24 h, or placebo (sodium chloride 0·9%). Patients, caregivers, and those assessing outcomes were masked to allocation. The primary outcome was death due to bleeding within 5 days of randomisation; analysis excluded patients who received neither dose of the allocated treatment and those for whom outcome data on death were unavailable. This trial was registered with Current Controlled Trials, ISRCTN11225767, and ClinicalTrials.gov, NCT01658124. Findings: Between July 4, 2013, and June 21, 2019, we randomly allocated 12 009 patients to receive tranexamic acid (5994, 49·9%) or matching placebo (6015, 50·1%), of whom 11 952 (99·5%) received the first dose of the allocated treatment. Death due to bleeding within 5 days of randomisation occurred in 222 (4%) of 5956 patients in the tranexamic acid group and in 226 (4%) of 5981 patients in the placebo group (risk ratio [RR] 0·99, 95% CI 0·82–1·18). Arterial thromboembolic events (myocardial infarction or stroke) were similar in the tranexamic acid group and placebo group (42 [0·7%] of 5952 vs 46 [0·8%] of 5977; 0·92; 0·60 to 1·39). Venous thromboembolic events (deep vein thrombosis or pulmonary embolism) were higher in tranexamic acid group than in the placebo group (48 [0·8%] of 5952 vs 26 [0·4%] of 5977; RR 1·85; 95% CI 1·15 to 2·98). Interpretation: We found that tranexamic acid did not reduce death from gastrointestinal bleeding. On the basis of our results, tranexamic acid should not be used for the treatment of gastrointestinal bleeding outside the context of a randomised trial