Myosin II filament dynamics in actin networks revealed with interferometric scattering microscopy

The plasma membrane and the underlying cytoskeletal cortex constitute active platforms for a variety of cellular processes. Recent work has shown that the remodeling acto-myosin network modifies local membrane organization, but the molecular details are only partly understood due to difficulties with experimentally accessing the relevant time and length scales. Here, we use interferometric scattering (iSCAT) microscopy to investigate a minimal acto-myosin network linked to a supported lipid bilayer membrane. Using the magnitude of the interferometric contrast, which is proportional to molecular mass, and fast acquisition rates, we detect, and image individual membrane attached actin filaments diffusing within the acto-myosin network and follow individual myosin II filament dynamics. We quantify myosin II filament dwell times and processivity as functions of ATP concentration, providing experimental evidence for the predicted ensemble behavior of myosin head domains. Our results show how decreasing ATP concentrations lead to both increasing dwell times of individual myosin II filaments and a global change from a remodeling to a contractile state of the acto-myosin network

Mass-sensitive particle tracking to elucidate the membrane-associated MinDE reaction cycle

An iSCAT image processing and analysis strategy enables mass-sensitive particle tracking (MSPT) of single unlabeled biomolecules on a supported lipid bilayer. MSPT was used to observe the (dis-)assembly of membrane complexes in real-time. In spite of their great importance in biology, methods providing access to spontaneous molecular interactions with and on biological membranes have been sparse. The recent advent of mass photometry to quantify mass distributions of unlabeled biomolecules landing on surfaces raised hopes that this approach could be transferred to membranes. Here, by introducing a new interferometric scattering (iSCAT) image processing and analysis strategy adapted to diffusing particles, we enable mass-sensitive particle tracking (MSPT) of single unlabeled biomolecules on a supported lipid bilayer. We applied this approach to the highly nonlinear reaction cycles underlying MinDE protein self-organization. MSPT allowed us to determine the stoichiometry and turnover of individual membrane-bound MinD/MinDE protein complexes and to quantify their size-dependent diffusion. This study demonstrates the potential of MSPT to enhance our quantitative understanding of membrane-associated biological systems.We thank D. Bollschweiler (Cryo-EM MPIB Core Facility) for the initial introduction to the commercial Refeyn OneMP mass photometer, the MPIB Biochemistry Core Facility (Recombinant Protein Production)

Disruptive Momentum: The Value of Implementing Best Practices in Health Research Postdoctoral Mentorship

COVID-19 called attention to the challenges postdoctoral fellows in health research face when they have times of prolonged disruption or changes in work conditions; this disruption revealed key insights on how mentors, fellows, and their institutions can work together to ensure training continuity. To prepare strong scientists, postdoctoral fellowships need mentoring, training, and networking opportunities to enhance fellows’ professional and skill development. In this article we outline potential solutions to minimize the impact of disruptions while promoting adaptable postdoctoral fellowship experiences by addressing how mentors and fellows alike can intervene on three key aspects of fellowships in health research: mentorship, training, and networking

Intrinsic Photoconductivity of Ultracold Fermions in Optical Lattices

We report on the experimental observation of an analog to a persistent alternating photocurrent in an ultracold gas of fermionic atoms in an optical lattice. The dynamics is induced and sustained by an external harmonic confinement. While particles in the excited band exhibit long-lived oscillations with a momentum dependent frequency a strikingly different behavior is observed for holes in the lowest band. An initial fast collapse is followed by subsequent periodic revivals. Both observations are fully explained by mapping the system onto a nonlinear pendulum.Comment: 5+7 pages, 4+4 figure

Alignment dependent chemisorption of vibrationally excited CH4(ν3) on Ni(100), Ni(110), and Ni(111)

International audienceWe present a stereodynamics study of the dissociative chemisorption of vibrationally excited methane on the (100), (110), and (111) planes of a nickel single crystal surface. Using linearly polarized infrared excitation of the antisymmetric C-H stretch normal mode vibration (ν3), we aligned the angular momentum and C-H stretch amplitude of CH4(ν3) in the laboratory frame and measured the alignment dependence of state-resolved reactivity of CH4 for the ν3 = 1, J = 0-3 quantum states over a range of incident translational energies. For all three surfaces studied, in-plane alignment of the C-H stretch results in the highest dissociation probability and alignment along the surface normal in the lowest reactivity. The largest alignment contrast between the maximum and minimum reactivity is observed for Ni(110), which has its surface atoms arranged in close-packed rows separated by one layer deep troughs. For Ni(110), we also probed for alignment effects relative to the direction of the Ni rows. In-plane C-H stretch alignment perpendicular to the surface rows results in higher reactivity than parallel to the surface rows. The alignment effects on Ni(110) and Ni(100) are independent of incident translational energy between 10 and 50 kJ/mol. Quantum state-resolved reaction probabilities are reported for CH4(ν3) on Ni(110) for translational energies between 10 and 50 kJ/mol

Quantum state specific reactant preparation in a molecular beam by rapid adiabatic passage

Highly efficient preparation of molecules in a specific rovibrationally excited state for gas/surface reactivity measurements is achieved in a molecular beam using tunable infrared (IR) radiation from a single mode continuous wave optical parametric oscillator (cw-OPO). We demonstrate that with appropriate focusing of the IR radiation, molecules in the molecular beam crossing the fixed frequency IR field experience a Doppler tuning that can be adjusted to achieve complete population inversion of a two-level system by rapid adiabatic passage (RAP). A room temperature pyroelectric detector is used to monitor the excited fraction in the molecular beam and the population inversion is detected and quantified using IR bleaching by a second IR-OPO. The second OPO is also used for complete population transfer to an overtone or combination vibration via double resonance excitation using two spatially separated RAP processes

Transformation Pathways of Silica under High Pressure

Concurrent molecular dynamics simulations and ab initio calculations show that densification of silica under pressure follows a ubiquitous two-stage mechanism. First, anions form a close-packed sub-lattice, governed by the strong repulsion between them. Next, cations redistribute onto the interstices. In cristobalite silica, the first stage is manifest by the formation of a metastable phase, which was observed experimentally a decade ago, but never indexed due to ambiguous diffraction patterns. Our simulations conclusively reveal its structure and its role in the densification of silica.Comment: 14 pages, 4 figure

Preoperative computed tomography staging of nonmetastatic colon cancer predicts outcome: implications for clinical trials

Colon cancer patients routinely undergo preoperative computed tomography (CT) scanning, but local staging is thought to be inaccurate. We aimed to determine if clinical outcome could be predicted from radiological features of the primary tumour. Consecutive patients at one hospital undergoing primary resection for colon cancer during 2000–2004 were included. Patients with visible metastases were excluded. Preoperative CT scans were reviewed independently by two radiologists blinded to histological stage and outcome. Images of the primary tumour were evaluated according to conventional TNM criteria and patients were stratified into ‘good' or ‘poor' prognosis groups. Comparison was made between prognostic group and actual clinical outcome. Hundred and twenty-six preoperative CT scans were reviewed. T-stage and nodal status was correctly predicted in only 60 and 62%, respectively. However, inter-observer agreement for prognostic group was 79% (κ=0.59) and 3-year relapse-free survival was 71 and 43% for the CT-predicted ‘good' and ‘poor' groups, respectively (P<0.0066). This compared favourably with 75 vs 43% for histology-predicted prognostic groups. Computed tomography is a robust method for stratifying patients preoperatively, with similar accuracy to histopathology for predicting outcome. Recognition of poor prognosis tumours preoperatively may permit investigation into the future use of neo-adjuvant therapy in colon cancer

Bad influence? – an investigation into the purported negative influence of foreign domestic helpers on children's second language English acquisition

This paper explores the purported negative influence of foreign domestic helpers (FDHs) on child second language acquisition (SLA) by studying Hong Kong Cantonese children's listening ability in second language (L2) English. 31 kindergarten third graders aged 4;6 to 6, and 29 first year secondary students aged 11-14 who have had a Filipino domestic helper at home took part in the study. In addition, 34 youngsters (20 in kindergarten, 14 in secondary) who did not have a Filipino helper participated as controls. Results from two listening tasks (picture choosing task, sound discrimination task) suggest that informants do not differ from the control in their abilities to listen to American-, British-, and Hong Kong English, and that they are better at listening to Filipino-accented English than the control. These findings cast doubts on the anecdotal belief of the harmful effect FDHs have on children's language acquisition including an L2. Moreover, the additional effect of being familiar with another variety of English is arguably a desirable outcome given that English is used as a lingua franca among non-native speakers on a daily basis in this highly globalised world