Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as of lipids in plasma membrane bioactivity. However, these dynamics can often only be ...visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, an extension of fluorescence correlation spectroscopy, FCS) on a super-resolution stimulated emission depletion (STED) microscope, we here extend previous observations of nanoscale lipid dynamics in the plasma membrane of living mammalian cells. STED-FLCS allows an improved determination of spatiotemporal heterogeneity in molecular diffusion and interaction dynamics via a novel gated detection scheme, as demonstrated by a comparison between STED-FLCS and previous conventional STED-FCS recordings on fluorescent phosphoglycerolipid and sphingolipid analogues in the plasma membrane of live mammalian cells. The STED-FLCS data indicate that biophysical and biochemical parameters such as the affinity for molecular complexes strongly change over space and time within a few seconds. Drug treatment for cholesterol depletion or actin cytoskeleton depolymerization not only results in the already previously observed decreased affinity for molecular interactions but also in a slight reduction of the spatiotemporal heterogeneity. STED-FLCS specifically demonstrates a significant improvement over previous gated STED-FCS experiments and with its improved spatial and temporal resolution is a novel tool for investigating how heterogeneities of the cellular plasma membrane may regulate biofunctionality.
Recently, following import by travel and migration, epidemic community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has caused nosocomial outbreaks in Europe, sometimes with a ...fatal outcome. We describe clinico-epidemiological characteristics of CA-MRSA detected by the European Network for the Surveillance of imported S. aureus (www.staphtrav.eu) from May 2011 to November 2016.
Sentinel surveillance at 13 travel clinics enrolling patients with travel-associated skin and soft-tissue infection (SSTI) and analysing lesion and nose swabs at one central laboratory.
A total of 564 independent case-patients with SSTI were enrolled and had 374 (67%) S. aureus-positive lesions, of which 14% (51/374) were MRSA. The majority of CA-MRSA isolates from SSTI were Panton–Valentine leucocidin (PVL) -positive (43/51, 84%). The risk of methicillin-resistance in imported S. aureus varied by travel region (p <0.001) and was highest in Latin America (16/57, 28%, 95% CI 17.0–41.5) and lowest in Sub-Saharan Africa (4/121, 3%, 95% CI 0.9–8.3). Major epidemic clones (USA300 / USA300 Latin-American Variant, Bengal Bay, South Pacific) accounted for more than one-third (19/51, 37%) of CA-MRSA imports. CA-MRSA SSTI in returnees was complicated (31/51 multiple lesions, 61%; 22/50 recurrences, 44%), led to health-care contact (22/51 surgical drainage, 43%; 7/50 hospitalization, 14%), was transmissible (13/47 reported similar SSTI in non-travelling contacts, 28%), and associated with S. aureus nasal colonization (28 of 51 CA-MRSA cases, 55%; 24 of 28 colonized with identical spa-type in nose and lesion, 85%).
Travel-associated CA-MRSA SSTI is a transmissible condition that leads to medical consultations and colonization of the infected host.
The particularly interdisciplinary nature of human microbiome research makes the organization and reporting of results spanning epidemiology, biology, bioinformatics, translational medicine and ...statistics a challenge. Commonly used reporting guidelines for observational or genetic epidemiology studies lack key features specific to microbiome studies. Therefore, a multidisciplinary group of microbiome epidemiology researchers adapted guidelines for observational and genetic studies to culture-independent human microbiome studies, and also developed new reporting elements for laboratory, bioinformatics and statistical analyses tailored to microbiome studies. The resulting tool, called 'Strengthening The Organization and Reporting of Microbiome Studies' (STORMS), is composed of a 17-item checklist organized into six sections that correspond to the typical sections of a scientific publication, presented as an editable table for inclusion in supplementary materials. The STORMS checklist provides guidance for concise and complete reporting of microbiome studies that will facilitate manuscript preparation, peer review, and reader comprehension of publications and comparative analysis of published results.
Unicompartmental knee arthroplasty (UKA) has become a standard procedure with good clinical outcome in patients with isolated medial osteoarthritis of the knee. However, the survival rates of UKA are ...still inferior compared to that of total knee arthroplasty. Aseptic loosening and wear are responsible for more than 50% of revisions. Therefore, this study evaluated the influence of the tibial slope on the wear rate in a medial UKA.
The wear rate of a medial mobile-bearing UKA (Univation® Aesculap, Tuttlingen, Deutschland) was evaluated according to the ISO 14243-1:2002(E) norm with a customized four-station servo-hydraulic knee wear simulator (EndoLab, Thansau, Germany). In the first group, the prostheses was medially implanted with 0° slope (n = 3) and in the second group the prostheses was medially implanted with 8° slope (n = 3). The lateral side was kept constant with 0° in both groups. For each implant, a total of 5.0 million cycles was performed and after every 0.5 million cycles the gravimetric wear rate was determined.
The wear rate in the 0° slope group was 3.46 ± 0.59 mg/million cycles and therefore significantly higher than in the 8° slope group with 0.99 ± 0.42 mg/million cycles (p < 0.01).
An increase in the tibial slope leads to a reduced wear rate in a mobile-bearing UKA. Therefore, at least for this mobile-bearing UKA a higher tibial slope seems favorable to reduce the wear. However, before an optimal position of the tibial slope can be recommended, further investigations are required to evaluate the influence of the tibial slope on other factors, such as the ligament tension or the strain on the lateral compartment.
Background The intrinsic healing capacity of articular cartilage remains poor, despite various attempts that have been undertaken to treat cartilage defects. This study describes the experimental use ...of a double-layer bioimplant consisting of a bone-substitute layer and a cartilage-substitute layer.
Animals and methods In group A, 12 implants were placed into osteochondral defects of the load-bearing area of rabbit femoral condyles. In group B, 12 implantations were done in the same manner, with a separating membrane consisting of cement between both layers to investigate ingrowth of mesenchymal stem cells from the subchondral marrow space. Group C, with 12 similar defects but without treatment, served as control. Investigations by microscopy and immunohistochemistry were done after 12 weeks.
Results All bioimplants showed coverage of the defect with a regeneration tissue that contained cartilage-like regions. Implants with a cement layer showed less cartilage and more fibrous tissue. The bioimplant group showed more cartilage-like regeneration tissue than the control groups, which only showed incomplete fibrous filling of the defects. Results from the second group supported the hypothesis that the subchondral space must be opened for adequate regeneration. Additional examinations were done using an established semiquantitative score. The bioimplant group showed a significant improvement in results compared to the group with the separating layer and the control group.
Interpretation Our findings indicate that cartilage repair by resorbable bioimplants seems to be a promising new approach, especially if mesenchymal stem cells are present and can differentiate under mechanical load.
Complex stellar winds from evolved stars
Stars less than eight times the mass of the Sun end their lives as planetary nebulae, structures of ionized gas thrown off by the star and heated by the ...exposed stellar core. Planetary nebulae are often bipolar in shape or contain complex morphological features such as rings or spirals. Decin
et al.
observed the stellar winds of 14 stars during their asymptotic giant branch (AGB) phase of stellar evolution, which immediately precedes the planetary nebula phase. They found morphologies in the AGB winds similar to planetary nebulae and demonstrated that they are produced by the influence of a binary companion on the AGB wind.
Science
, this issue p.
1497
Complex morphologies in the winds of asymptotic giant branch stars and planetary nebulae are produced by binary interactions.
Binary interactions dominate the evolution of massive stars, but their role is less clear for low- and intermediate-mass stars. The evolution of a spherical wind from an asymptotic giant branch (AGB) star into a nonspherical planetary nebula (PN) could be due to binary interactions. We observed a sample of AGB stars with the Atacama Large Millimeter/submillimeter Array (ALMA) and found that their winds exhibit distinct nonspherical geometries with morphological similarities to planetary nebulae (PNe). We infer that the same physics shapes both AGB winds and PNe; additionally, the morphology and AGB mass-loss rate are correlated. These characteristics can be explained by binary interaction. We propose an evolutionary scenario for AGB morphologies that is consistent with observed phenomena in AGB stars and PNe.
How predictable are life trajectories? We investigated this question with a scientific mass collaboration using the common task method; 160 teams built predictive models for six life outcomes using ...data from the Fragile Families and Child Wellbeing Study, a high-quality birth cohort study. Despite using a rich dataset and applying machine-learning methods optimized for prediction, the best predictions were not very accurate and were only slightly better than those from a simple benchmark model. Within each outcome, prediction error was strongly associated with the family being predicted and weakly associated with the technique used to generate the prediction. Overall, these results suggest practical limits to the predictability of life outcomes in some settings and illustrate the value of mass collaborations in the social sciences.
Besides its established functions in intermediary metabolism and developmental processes, the nuclear receptor peroxisome proliferator-activated receptor β/δ (PPARβ/δ) has a less defined role in ...tumorigenesis. In the present study, we have identified a function for PPARβ/δ in cancer cell invasion. We show that two structurally divergent inhibitory ligands for PPARβ/δ, the inverse agonists ST247 and DG172, strongly inhibit the serum- and transforming growth factor β (TGFβ)-induced invasion of MDA-MB-231 human breast cancer cells into a three-dimensional matrigel matrix. To elucidate the molecular basis of this finding, we performed chromatin immunoprecipitation sequencing (ChIP-Seq) and microarray analyses, which identified the gene encoding angiopoietin-like 4 (ANGPTL4) as the major transcriptional PPARβ/δ target in MDA-MB-231 cells, previously implicated in TGFβ-mediated tumor progression and metastatic dissemination. We show that the induction of ANGPTL4 by TGFβ and other oncogenic signals is strongly repressed by ST247 and DG172 in a PPARβ/δ-dependent fashion, resulting in the inhibition of ANGPTL4 secretion. This effect is attributable to these ligands' ability to induce a dominant transcriptional repressor complex at the site of transcription initiation that blocks preinitiation complex formation through an histone deacetylase-independent, non-canonical mechanism. Repression of ANGPTL4 transcription by inverse PPARβ/δ agonists is functionally linked to the inhibition of cancer cell invasion into a three-dimensional matrix, as (i) invasion of MDA-MB-231 cells is critically dependent on ANGPTL4 expression, (ii) recombinant ANGPTL4 stimulates invasion, and (iii) reverses the inhibitory effect of ST247 and DG172. These findings indicate that a PPARβ/δ-ANGPTL4 pathway is involved in the regulation of tumor cell invasion and that its pharmacological manipulation by inverse PPARβ/δ agonists is feasible.
•We recruited a large sample (n=1181) of participants using crowdsourcing technology.•We assessed discount rates for delayed, probabilistic, and social monetary rewards.•Delay and social discounting ...rates were higher in obese than non-obese individuals.•Probability discounting rates were comparable in obese than non-obese individuals.
Previous research comparing obese and non-obese samples on the delayed discounting procedure has produced mixed results. The aim of the current study was to clarify these discrepant findings by comparing a variety of temporal discounting measures in a large sample of internet users (n=1163) obtained from a crowdsourcing service, Amazon Mechanical Turk (AMT). Measures of temporal, social–temporal (a combination of standard and social temporal), and probability discounting were obtained. Significant differences were obtained on all discounting measures except probability discounting, but the obtained effect sizes were small. These data suggest that larger-N studies will be more likely to detect differences between obese and non-obese samples, and may afford the opportunity, in future studies, to decompose a large obese sample into different subgroups to examine the effect of other relevant measures, such as the reinforcing value of food, on discounting.
Small-molecule inhibitors that target bromodomains outside of the bromodomain and extra-terminal (BET) sub-family are lacking. Here, we describe highly potent and selective ligands for the ...bromodomain module of the human lysine acetyl transferase CBP/p300, developed from a series of 5-isoxazolyl-benzimidazoles. Our starting point was a fragment hit, which was optimized into a more potent and selective lead using parallel synthesis employing Suzuki couplings, benzimidazole-forming reactions, and reductive aminations. The selectivity of the lead compound against other bromodomain family members was investigated using a thermal stability assay, which revealed some inhibition of the structurally related BET family members. To address the BET selectivity issue, X-ray crystal structures of the lead compound bound to the CREB binding protein (CBP) and the first bromodomain of BRD4 (BRD4(1)) were used to guide the design of more selective compounds. The crystal structures obtained revealed two distinct binding modes. By varying the aryl substitution pattern and developing conformationally constrained analogues, selectivity for CBP over BRD4(1) was increased. The optimized compound is highly potent (K d = 21 nM) and selective, displaying 40-fold selectivity over BRD4(1). Cellular activity was demonstrated using fluorescence recovery after photo-bleaching (FRAP) and a p53 reporter assay. The optimized compounds are cell-active and have nanomolar affinity for CBP/p300; therefore, they should be useful in studies investigating the biological roles of CBP and p300 and to validate the CBP and p300 bromodomains as therapeutic targets.
PDF
