Protein Reconstitution Inside Giant Unilamellar Vesicles

2021 ◽  
Vol 50 (1) ◽  
Author(s):  
Thomas Litschel ◽  
Petra Schwille

Giant unilamellar vesicles (GUVs) have gained great popularity as mimicries for cellular membranes. As their sizes are comfortably above the optical resolution limit, and their lipid composition is easily controlled, they are ideal for quantitative light microscopic investigation of dynamic processes in and on membranes. However, reconstitution of functional proteins into the lumen or the GUV membrane itself has proven technically challenging. In recent years, a selection of techniques has been introduced that tremendously improve GUV-assay development and enable the precise investigation of protein–membrane interactions under well-controlled conditions. Moreover, due to these methodological advances, GUVs are considered important candidates as protocells in bottom-up synthetic biology. In this review, we discuss the state of the art of the most important vesicle production and protein encapsulation methods and highlight some key protein systems whose functional reconstitution has advanced the field. Expected final online publication date for the Annual Review of Biophysics, Volume 50 is May 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Author(s):  
Patrick D. McGorry ◽  
Cristina Mei

Current silo-based diagnostic systems for mental disorders lack utility and fail to fulfil a fundamental purpose of diagnosis: to guide treatment planning and predict outcomes. Diagnostic reform has gained momentum, and clinical staging has emerged as a promising framework to improve the precision of diagnosis, particularly in early illness stages, and fill current gaps in linking diagnosis to more personalized and effective intervention, prognosis, and neurobiological markers. Transdiagnostic clinical staging recognizes that the early development of mental ill-health is marked by substantial fluidity and that symptoms may, although not inevitably, evolve into more stable diagnosable syndromes. Staging facilitates the selection of interventions that are proportionate to the current need and risk of illness progression and provides an efficient framework to organize biomarker data and guide service delivery. Here, we provide an overview of transdiagnostic clinical staging and summarize key evidence supporting its ability to integrate biomarkers and guide mental health care. Expected final online publication date for the Annual Review of Developmental Psychology, Volume 3 is December 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


2020 ◽  
Vol 7 (1) ◽  
Author(s):  
Roberta D'Alessandro

Language change as a result of language contact is studied in many different ways using a number of different methodologies. This article provides an overview of the main approaches to syntactic change in contact (CIC), focusing on the Romance language group. Romance languages are widely documented both synchronically and diachronically. They have been in extensive contact with other language families both in bilingual contexts and in creolization contexts. Furthermore, they present great microvariation. They are therefore ideal to tackle language change in contact. Given the breadth of studies targeting Romance languages in contact, only a selection of facts is considered here, namely pro-drop, differential object marking (DOM), and deixis. The article shows that microcontact, i.e., contact between minimally different grammars, is a necessary dimension to be considered within contact studies, as it provides insights that are often radically different from those provided by the observation of contact between maximally different languages. Expected final online publication date for the Annual Review of Linguistics, Volume 7 is January 14, 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


2022 ◽  
Vol 45 (1) ◽  
Author(s):  
Roberto Leiras ◽  
Jared M. Cregg ◽  
Ole Kiehn

Locomotion is a universal motor behavior that is expressed as the output of many integrated brain functions. Locomotion is organized at several levels of the nervous system, with brainstem circuits acting as the gate between brain areas regulating innate, emotional, or motivational locomotion and executive spinal circuits. Here we review recent advances on brainstem circuits involved in controlling locomotion. We describe how delineated command circuits govern the start, speed, stop, and steering of locomotion. We also discuss how these pathways interface between executive circuits in the spinal cord and diverse brain areas important for context-specific selection of locomotion. A recurrent theme is the need to establish a functional connectome to and from brainstem command circuits. Finally, we point to unresolved issues concerning the integrated function of locomotor control. Expected final online publication date for the Annual Review of Neuroscience, Volume 45 is July 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


Author(s):  
Laurent Bouffier ◽  
Dodzi Zigah ◽  
Neso Sojic ◽  
Alexander Kuhn

This contribution reviews a selection of the most recent studies on the use of bipolar electrochemistry in the framework of analytical chemistry. Despite the fact that the concept is not new, with several important studies dating back to the middle of the last century, completely novel and very original approaches have emerged over the last decade. This current revival illustrates that scientists still (re)discover some exciting virtues of this approach, which are useful in many different areas, especially for tackling analytical challenges in an unconventional way. In several cases, this “wireless” electrochemistry strategy enables carrying out measurements that are simply not possible with classic electrochemical approaches. This review will hopefully stimulate new ideas and trigger scientists to integrate some aspects of bipolar electrochemistry in their work in order to drive the topic into yet unexplored and eventually completely unexpected directions. Expected final online publication date for the Annual Review of Analytical Chemistry, Volume 14 is June 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


Author(s):  
Yoav Weizman ◽  
Ezra Baruch

Abstract In recent years, two new techniques were introduced for flip chip debug; the Laser Voltage Probing (LVP) technique and Time Resolved Light Emission Microscopy (TRLEM). Both techniques utilize the silicon’s relative transparency to wavelengths longer than the band gap. This inherent wavelength limitation, together with the shrinking dimensions of modern CMOS devices, limit the capabilities of these tools. It is known that the optical resolution limits of the LVP and TRLEM techniques are bounded by the diffraction limit which is ~1um for both tools using standard optics. This limitation was reduced with the addition of immersion lens optics. Nevertheless, even with this improvement, shrinking transistor geometry is leading to increased acquisition time, and the overlapping effect between adjacent nodes remains a critical issue. The resolution limit is an order of magnitude above the device feature densities in the < 90nm era. The scaling down of transistor geometry is leading to the inevitable consequence where more than 50% of the transistors in 90nm process have widths smaller than 0.4um. The acquisition time of such nodes becomes unreasonably long. In order to examine nodes in a dense logic cuicuit, cross talk and convolution effects between neighboring signals also need to be considered. In this paper we will demonstrate the impact that these effects may have on modern design. In order to maintain the debug capability, with the currently available analytical tools for future technologies, conceptual modification of the FA process is required. This process should start on the IC design board where the VLSI designer should be familiar with FA constraints, and thus apply features that will enable enhanced FA capabilities to the circuit in hand during the electrical design or during the physical design stages. The necessity for reliable failure analysis in real-time should dictate that the designer of advanced VLSI blocks incorporates failure analysis constraints among other design rules. The purpose of this research is to supply the scientific basis for the optimal incorporation of design rules for optical probing in the < 90nm gate era. Circuit designers are usually familiar with the nodes in the design which are critical for debug, and the type of measurement (logic or DC level) they require. The designer should enable the measurement of these signals by applying certain circuit and physical constraints. The implementation of these constraints may be done at the cell level, the block level or during the integration. We will discuss the solutions, which should be considered in order to mitigate tool limitations, and also to enable their use for next generation processes.


Life ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 634
Author(s):  
Ylenia Miele ◽  
Gábor Holló ◽  
István Lagzi ◽  
Federico Rossi

The budding and division of artificial cells engineered from vesicles and droplets have gained much attention in the past few decades due to an increased interest in designing stimuli-responsive synthetic systems. Proper control of the division process is one of the main challenges in the field of synthetic biology and, especially in the context of the origin of life studies, it would be helpful to look for the simplest chemical and physical processes likely at play in prebiotic conditions. Here we show that pH-sensitive giant unilamellar vesicles composed of mixed phospholipid/fatty acid membranes undergo a budding process, internally fuelled by the urea–urease enzymatic reaction, only for a given range of the membrane composition. A gentle interplay between the effects of the membrane composition on the elasticity and the preferred area difference of the bilayer is responsible for the existence of a narrow range of membrane composition yielding a high probability for budding of the vesicles.


Langmuir ◽  
2021 ◽  
Vol 37 (3) ◽  
pp. 1082-1088
Author(s):  
Chiho Kataoka-Hamai ◽  
Kohsaku Kawakami

Soft Matter ◽  
2021 ◽  
Author(s):  
Peng Bao ◽  
Daniel A. Paterson ◽  
Sally A. Peyman ◽  
J. Cliff Jones ◽  
Jonathan A. T. Sandoe ◽  
...  

We describe a modified microfluidic method for making Giant Unilamellar Vesicles (GUVs) via water/octanol-lipid/water double emulsion droplets and encapsulation of nematic lyotropic liquid crystals (LNLCs).


Author(s):  
Elliott S. Chiu ◽  
Sue VandeWoude

Endogenous retroviruses (ERVs) serve as markers of ancient viral infections and provide invaluable insight into host and viral evolution. ERVs have been exapted to assist in performing basic biological functions, including placentation, immune modulation, and oncogenesis. A subset of ERVs share high nucleotide similarity to circulating horizontally transmitted exogenous retrovirus (XRV) progenitors. In these cases, ERV–XRV interactions have been documented and include ( a) recombination to result in ERV–XRV chimeras, ( b) ERV induction of immune self-tolerance to XRV antigens, ( c) ERV antigen interference with XRV receptor binding, and ( d) interactions resulting in both enhancement and restriction of XRV infections. Whereas the mechanisms governing recombination and immune self-tolerance have been partially determined, enhancement and restriction of XRV infection are virus specific and only partially understood. This review summarizes interactions between six unique ERV–XRV pairs, highlighting important ERV biological functions and potential evolutionary histories in vertebrate hosts. Expected final online publication date for the Annual Review of Animal Biosciences, Volume 9 is February 16, 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.


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