scholarly journals TMAO regulates the rigidity of kinesin-propelled microtubules

2021 ◽  
Author(s):  
Tasrina Munmun ◽  
Arif Md. Rashedul Kabir ◽  
Kazuki Sada ◽  
Akira Kakugo
Keyword(s):  
Deep Sea ◽  
Materials Science ◽  
Persistence Length ◽  
Simple Strategy ◽  
Biomolecular Motors ◽  
Wide Range

AbstractWe demonstrate that the rigidity of the microtubules (MTs), propelled by kinesins in an in vitro gliding assay, can be modulated using the deep-sea osmolyte trimethylamine N-oxide (TMAO). By varying the concentration of TMAO in the gliding assay, the rigidity of the MTs is modulated over a wide range. By employing this approach, we are able to reduce the persistence length of MTs, a measure of MT rigidity, ∼8 fold using TMAO of the concentration of 1.5 M. The rigidity of gliding MTs can be restored by eliminating the TMAO from the gliding assay. This work offers a simple strategy to regulate the rigidity of kinesin-propelled MTs in situ and would widen the applications of biomolecular motors in nanotechnology, materials science, and bioengineering.

scholarly journals Quantification of assembly forces during creation of head-neck taper junction considering soft tissue bearing: a biomechanical study

Arthroplasty ◽  
2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Toni Wendler ◽  
Torsten Prietzel ◽  
Robert Möbius ◽  
Jean-Pierre Fischer ◽  
Andreas Roth ◽  
...  
Keyword(s):  
Soft Tissue ◽  
Work Experience ◽  
Soft Tissues ◽  
Force Sensor ◽  
Biomechanical Study ◽  
Measuring System ◽  
Wide Range ◽  
Head Neck

Abstract Background All current total hip arthroplasty (THA) systems are modular in design. Only during the operation femoral head and stem get connected by a Morse taper junction. The junction is realized by hammer blows from the surgeon. Decisive for the junction strength is the maximum force acting once in the direction of the neck axis, which is mainly influenced by the applied impulse and surrounding soft tissues. This leads to large differences in assembly forces between the surgeries. This study aimed to quantify the assembly forces of different surgeons under influence of surrounding soft tissue. Methods First, a measuring system, consisting of a prosthesis and a hammer, was developed. Both components are equipped with a piezoelectric force sensor. Initially, in situ experiments on human cadavers were carried out using this system in order to determine the actual assembly forces and to characterize the influence of human soft tissues. Afterwards, an in vitro model in the form of an artificial femur (Sawbones Europe AB, Malmo, Sweden) with implanted measuring stem embedded in gelatine was developed. The gelatine mixture was chosen in such a way that assembly forces applied to the model corresponded to those in situ. A study involving 31 surgeons was carried out on the aforementioned in vitro model, in which the assembly forces were determined. Results A model was developed, with the influence of human soft tissues being taken into account. The assembly forces measured on the in vitro model were, on average, 2037.2 N ± 724.9 N, ranging from 822.5 N to 3835.2 N. The comparison among the surgeons showed no significant differences in sex (P = 0.09), work experience (P = 0.71) and number of THAs performed per year (P = 0.69). Conclusions All measured assembly forces were below 4 kN, which is recommended in the literature. This could lead to increased corrosion following fretting in the head-neck interface. In addition, there was a very wide range of assembly forces among the surgeons, although other influencing factors such as different implant sizes or materials were not taken into account. To ensure optimal assembly force, the impaction should be standardized, e.g., by using an appropriate surgical instrument.

scholarly journals Construction of a mammalian embryo model from stem cells organized by a morphogen signalling centre

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Peng-Fei Xu ◽  
Ricardo Moraes Borges ◽  
Jonathan Fillatre ◽  
Maraysa de Oliveira-Melo ◽  
Tao Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Tracking ◽  
Cardiac Tissue ◽  
Embryonic Stem ◽  
Mammalian Embryo ◽  
Wide Range ◽  
Vitro Model ◽  
Neurula Stage

AbstractGenerating properly differentiated embryonic structures in vitro from pluripotent stem cells remains a challenge. Here we show that instruction of aggregates of mouse embryonic stem cells with an experimentally engineered morphogen signalling centre, that functions as an organizer, results in the development of embryo-like entities (embryoids). In situ hybridization, immunolabelling, cell tracking and transcriptomic analyses show that these embryoids form the three germ layers through a gastrulation process and that they exhibit a wide range of developmental structures, highly similar to neurula-stage mouse embryos. Embryoids are organized around an axial chordamesoderm, with a dorsal neural plate that displays histological properties similar to the murine embryo neuroepithelium and that folds into a neural tube patterned antero-posteriorly from the posterior midbrain to the tip of the tail. Lateral to the chordamesoderm, embryoids display somitic and intermediate mesoderm, with beating cardiac tissue anteriorly and formation of a vasculature network. Ventrally, embryoids differentiate a primitive gut tube, which is patterned both antero-posteriorly and dorso-ventrally. Altogether, embryoids provide an in vitro model of mammalian embryo that displays extensive development of germ layer derivatives and that promises to be a powerful tool for in vitro studies and disease modelling.

Minimum intracellular PO2 for maximum cytochrome turnover in red muscle in situ

1987 ◽  
Vol 252 (5) ◽  
pp. H906-H915 ◽  
Author(s):  
T. E. Gayeski ◽  
R. J. Connett ◽  
C. R. Honig
Keyword(s):  
Energy Demand ◽  
Concentration Ratio ◽  
Probability Distributions ◽  
Michaelis Constant ◽  
Cytochrome Aa3 ◽  
Wide Range ◽  
Red Muscle

Probability distributions of myoglobin (Mb) saturation and intracellular PO2 were determined with subcellular spatial resolution in dog gracilis muscles during steady-state twitch contraction at 5-100% of maximal rate of O2 consumption (VO2). Calculations (Clark, A., and P. A.A. Clark. Biophys. J. 48: 931-938, 1985) and measurements (Gayeski, T. E. J., and C. R. Honig. Adv. Exp. Med. Biol. 200: 487-494, 1986) indicate that the PO2 in equilibrium with Mb is virtually identical to the PO2 at cytochrome aa3. Median intracellular PO2 and PO2 in the lower tails of probability distributions were poorly correlated with VO2. The variability of cell PO2 was greatly diminished when median PO2 was less than the PO2 for half saturation of MB, since Mb acts as a PO2 buffer. The lower tails of PO2 distributions contained almost no anoxic loci even when median PO2 was less than 1 Torr. VO2 was well correlated with the concentration ratio of phosphocreatine to free creatine (PCr/Crf) over a wide range of PO2. PO2 greater than or equal to 0.5 Torr supported maximal VO2 and energy demand. We conclude that 1) the mechanism of action of cytochrome aa3 is the same in red muscle in vivo as in mitochondria in vitro, and 2) an upper bound on the apparent Michaelis constant for maximal VO2 of red muscle is approximately 0.06 Torr.

In vitro techniques to replace in vivo methods for estimating amino acid supply

1998 ◽  
Vol 22 ◽  
pp. 131-144 ◽  
Author(s):  
T. Hvelplund ◽  
M. R. Weisbjerg
Keyword(s):  
Protein Synthesis ◽  
Alternative Methods ◽  
Microbial Protein ◽  
Microbial Protein Synthesis ◽  
Wide Range ◽  
Intestinal Digestibility ◽  
Protein Degradability

Abstract Expressing the protein value of a food involves measurements of several of its characteristics. Many in vivo studies have shown, that the protein degradability in the rumen varies substantially both between and within foods and therefore estimation of protein degradability in the rumen is an important task in protein evaluation. The most common method used has been the in situ (in sacco, nylon bag) method but many in vitro methods have been introduced and are based on use of either buffer solubility, chemical methods, rumen fluid or enzymes. None of these in vitro methods has proven to be of general use. In further development of in vitro methods as well as the in situ method a major problem is lack of a set of samples with a ‘true’ in vivo degradability which can be used for calibration of alternative methods. Microbial protein synthesis in the rumen has to be related to food characteristics which can be analysed easily. In vitro methods which can predict organic matter digestibility in foods are available and can be used to predict microbial protein synthesis in the rumen. Intestinal digestibility of undegraded dietary protein varies substantially both between and within foods and easy methods to estimate intestinal digestibility are therefore essential. The mobile bag method is easy to use and seems to give reliable results on most foods but requires access to duodenal cannulated animals which prevents this method from being routine. Alternative in vitro methods have been developed but further research is required for validation of these methods on a wide range of foods before they can be accepted for general use.

scholarly journals Survival of and In Situ Gene Expression by Vibrio vulnificus at Varying Salinities in Estuarine Environments

2007 ◽  
Vol 74 (1) ◽  
pp. 182-187 ◽  
Author(s):  
Melissa K. Jones ◽  
Elizabeth Warner ◽  
James D. Oliver
Keyword(s):  
Gene Expression ◽  
Vibrio Vulnificus ◽  
Natural Habitat ◽  
Term Survival ◽  
Long Term Survival ◽  
Wide Range ◽  
Opportunistic Human Pathogen

ABSTRACT The opportunistic human pathogen Vibrio vulnificus survives in a wide range of ecological environments, which demonstrates its ability to adapt to highly variable conditions. Survival and gene expression under various conditions have been extensively studied in vitro; however, little work has been done to evaluate this bacterium in its natural habitat. Therefore, this study monitored the long-term survival of V. vulnificus in situ and simultaneously evaluated the expression of stress (rpoS, relA, hfq, and groEL) and putative virulence (vvpE, smcR, viuB, and trkA) genes at estuarine sites of varying salinity. Additionally, the survival and gene expression of an rpoS and an oxyR mutant were examined under the same conditions. Differences between the sampling sites in the long-term survival of any strain were not seen. However, differences were seen in the expression of viuB, trkA, and relA but our findings differed from what has been previously shown in vitro. These results also routinely demonstrated that genes required for survival under in vitro stress or host conditions are not necessarily required for survival in the water column. Overall, this study highlights the need for further in situ evaluation of this bacterium in order to gain a true understanding of its ecology and how it relates to its natural habitat.

Thermal sensitivity of mitochondrial functions in permeabilized muscle fibers from two populations of Drosophila simulans with divergent mitotypes

2011 ◽  
Vol 301 (1) ◽  
pp. R48-R59 ◽  
Author(s):  
Nicolas Pichaud ◽  
J. William O. Ballard ◽  
Robert M. Tanguay ◽  
Pierre U. Blier
Keyword(s):  
Thermal Sensitivity ◽  
In Vitro Study ◽  
Drosophila Simulans ◽  
External Temperature ◽  
Aerobic Activity ◽  
Mitochondrial Functions ◽  
Wide Range ◽  
Catalytic Capacity

In ectotherms, the external temperature is experienced by the mitochondria, and the mitochondrial respiration of different genotypes is likely to change as a result. Using high-resolution respirometry with permeabilized fibers (an in situ approach), we tried to identify differences in mitochondrial performance and thermal sensitivity of two Drosophila simulans populations with two different mitochondrial types ( siII and siIII) and geographical distributions. Maximal state 3 respiration rates obtained with electrons converging at the Q junction of the electron transport system (ETS) differed between the mitotypes at 24°C. Catalytic capacities were higher in flies harboring siII than in those harboring siIII mitochondrial DNA (2,129 vs. 1,390 pmol O2·s−1·mg protein−1). The cytochrome c oxidase activity was also higher in siII than siIII flies (3,712 vs. 2,688 pmol O2·s−1·mg protein−1). The higher catalytic capacity detected in the siII mitotype could provide an advantage in terms of intensity of aerobic activity, endurance, or both, if the intensity of exercise that can be aerobically performed is partly dictated by the aerobic capacity of the tissue. Moreover, thermal sensitivity results showed that even if temperature affects the catalytic capacity of the different enzymes of the ETS, both mitotypes revealed high tolerance to temperature variation. Previous in vitro study failed to detect any consistent functional mitochondrial differences between the same mitotypes. We conclude that the in situ approach is more sensitive and that the ETS is a robust system in terms of functional and regulatory properties across a wide range of temperatures.

scholarly journals Preparation, Characterization, and Optimization of an In Vitro C30 Carotenoid Pathway

2005 ◽  
Vol 71 (11) ◽  
pp. 6578-6583 ◽  
Author(s):  
Bosung Ku ◽  
Jae-Cheol Jeong ◽  
Benjamin N. Mijts ◽  
Claudia Schmidt-Dannert ◽  
Jonathan S. Dordick
Keyword(s):  
Phase System ◽  
Reaction Conditions ◽  
Two Phase ◽  
Gene Encoding ◽  
In Situ Extraction ◽  
Steady State Kinetics ◽  
Wide Range ◽  
Carotenoid Synthesis

ABSTRACT The ispA gene encoding farnesyl pyrophosphate (FPP) synthase from Escherichia coli and the crtM gene encoding 4,4′-diapophytoene (DAP) synthase from Staphylococcus aureus were overexpressed and purified for use in vitro. Steady-state kinetics for FPP synthase and DAP synthase, individually and in sequence, were determined under optimized reaction conditions. For the two-step reaction, the DAP product was unstable in aqueous buffer; however, in situ extraction using an aqueous-organic two-phase system resulted in a 100% conversion of isopentenyl pyrophosphate and dimethylallyl pyrophosphate into DAP. This aqueous-organic two-phase system is the first demonstration of an in vitro carotenoid synthesis pathway performed with in situ extraction, which enables quantitative conversions. This approach, if extended to a wide range of isoprenoid-based pathways, could lead to the synthesis of novel carotenoids and their derivatives.

Self-Lubrication of Machining Tools in Dry via In-situ Formed Tribofilms

2005 ◽  
Vol 890 ◽  
Author(s):  
Tatsuhiko Aizawa ◽  
Shigeo Yamamoto ◽  
Taro Sumitomo
Keyword(s):  
Materials Science ◽  
Protective Coatings ◽  
Cutting Speed ◽  
Green Manufacturing ◽  
Dry Machining ◽  
Wide Range ◽  
In Situ Formation ◽  
Green Manufacturing Technology ◽  
Turning Test

ABSTRACTGreen manufacturing technology has become a significant innovative keyword to reduce the environmental burden and CO2 emission at the stage of manufacturing and production. In daily operations by machining, huge amount of lubricating oils and cleansing agents is wasted so that dry machining technology provides us a way to completely reduce these wastes. Among several candidates, self-lubrication via in-situ formation of lubricious oxide films is accommodated to protective coatings in order to attain low friction and wear state even at higher cutting speed range. Materials science of in-situ formed lubricious oxide tribofilms is stated with consideration of accommodation mechanism via the chlorine implantation. Mechanical characterization is made for evaluation on elasto-plastic deformation of lubricious oxides. Turning test is employed to evaluate dry machinability of various tools for wide range of cutting speed. Self-lubrication in dry machining is described both for bare WC and TiCN-coated WC tools with and without chlorine implantation. Precise microstructure analyses are made by using the laser microscope, EDS and XPS. In-situ formation of lubricous oxides proves that self-lubrication process takes place even in dry machining to reduce the flank wear and friction coefficient.

Aberration correction: zooming out to overview

2009 ◽  
Vol 367 (1903) ◽  
pp. 3859-3870 ◽  
Author(s):  
A. Howie
Keyword(s):  
Materials Science ◽  
Complex Structure ◽  
Chromatic Aberration ◽  
Aberration Correction ◽  
Contrast Imaging ◽  
Atomic Column ◽  
Wide Range ◽  
The Many

In the structural characterization of thin specimens by projection (atomic column) imaging, the revolutionary development of aberration-corrected electron microscopy has already brought significant improvements not only in spatial resolution but also in improved image contrast. Some highlights from the symposium are summarized. Despite the purchasing and operating costs as well as the demands they place on operator skills, a staggering number of these new microscopes has already been installed worldwide. Serious challenges, therefore, arise including the need to attract customers from a wide range of disciplines where complex structure problems may require the development of new imaging modes. The ability to image at large scattering angles may be useful in mitigating some of the many as-yet uncorrected delocalization mechanisms that then arise and are systematically identified here. Larger specimen volumes made possible by chromatic aberration correction will benefit the development of more relevant in situ observations, particularly in materials science and catalysis. With additional incorporation of phase shifting electrodes or other devices, these chromatic aberration correctors could also be important for efficient phase contrast imaging in easily damaged biological structures. For many of these formidable problems, earlier experience of the optical microscopy community may teach some lessons.

scholarly journals A versatile nanoreactor for complementary in situ X-ray and electron microscopy studies in catalysis and materials science

2019 ◽  
Vol 26 (5) ◽  
pp. 1769-1781 ◽  
Author(s):  
Yakub Fam ◽  
Thomas L. Sheppard ◽  
Johannes Becher ◽  
Dennis Scherhaufer ◽  
Heinz Lambach ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Materials Science ◽  
Functional Materials ◽  
Ambient Conditions ◽  
Zeolite Sample ◽  
X Ray ◽  
Fluorescence Measurements ◽  
Wide Range ◽  
Modular Platform

Two in situ `nanoreactors' for high-resolution imaging of catalysts have been designed and applied at the hard X-ray nanoprobe endstation at beamline P06 of the PETRA III synchrotron radiation source. The reactors house samples supported on commercial MEMS chips, and were applied for complementary hard X-ray ptychography (23 nm spatial resolution) and transmission electron microscopy, with additional X-ray fluorescence measurements. The reactors allow pressures of 100 kPa and temperatures of up to 1573 K, offering a wide range of conditions relevant for catalysis. Ptychographic tomography was demonstrated at limited tilting angles of at least ±35° within the reactors and ±65° on the naked sample holders. Two case studies were selected to demonstrate the functionality of the reactors: (i) annealing of hierarchical nanoporous gold up to 923 K under inert He environment and (ii) acquisition of a ptychographic projection series at ±35° of a hierarchically structured macroporous zeolite sample under ambient conditions. The reactors are shown to be a flexible and modular platform for in situ studies in catalysis and materials science which may be adapted for a range of sample and experiment types, opening new characterization pathways in correlative multimodal in situ analysis of functional materials at work. The cells will presently be made available for all interested users of beamline P06 at PETRA III.

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