Microtubules regulate cardiomyocyte transversal Young's modulus.

The field of cardiomyocyte mechanobiology is gaining significant attention, due to accumulating evidence concerning the significant role of cellular mechanical effects on the integrated function of the heart. To date, the protein titin has been demonstrated as a major contributor to the cardiomyocytes Young's modulus (YM). The microtubular network represents another potential regulator of cardiac mechanics. However, the contribution of microtubules (MTs) to the membrane YM is still understudied and has not been interrogated in the context of myocardial infarction (MI) or mechanical loading and unloading. Using nanoscale mechanoscanning ion conductance microscopy, we demonstrate that MTs contribute to cardiomyocyte transverse YM in healthy and pathological states with different mechanical loading. Specifically, we show that posttranslational modifications of MTs have differing effects on cardiomyocyte YM: Acetylation provides flexibility, whereas detyrosination imparts rigidity. Further studies demonstrate that there is no correlation between the total protein amount of acetylated and detyrosinated MT. Yet, in the polymerized-only populations, an increased level of acetylation results in a decline of detyrosinated MTs in an MI model.

Rapid formation of human immunodeficiency virus-like particles.

Understanding the molecular mechanisms involved in the assembly of viruses is essential for discerning how viruses transmit from cell to cell and host to host. Although molecular aspects of assembly have been studied for many viruses, we still have little information about these events in real time. Enveloped viruses such as HIV that assemble at, and bud from, the plasma membrane have been studied in some detail using live cell fluorescence imaging techniques; however, these approaches provide little information about the real-time morphological changes that take place as viral components come together to form individual virus particles. Here we used correlative scanning ion conductance microscopy and fluorescence confocal microscopy to measure the topological changes, together with the recruitment of fluorescently labeled viral proteins such as Gag and Vpr, during the assembly and release of individual HIV virus-like particles (VLPs) from the top, nonadherent surfaces of living cells. We show that 1) labeling of viral proteins with green fluorescent protein affects particle formation, 2) the kinetics of particle assembly on different plasma membrane domains can vary, possibly as a consequence of differences in membrane biophysical properties, and 3) VLPs budding from the top, unimpeded surface of cells can reach full size in 20 s and disappear from the budding site in 0.5 to 3 min from the moment curvature is initially detected, significantly faster than has been previously reported.

Stigmatic optical system with corrected third-order spherical aberration for an arbitrary position of the object.

Our paper is focused on a problem of analysis and design of a stigmatic optical system that has corrected third-order spherical aberration for an arbitrary position of the object. The relations for Seidel aberration coefficients of the system for an object at infinity that must be satisfied to ensure that the third-order spherical aberration does not depend on the position of the object are given. The method for obtaining design parameters of the initial optical system that can serve as a good starting point for further refinement using numerical optimization methods is proposed. Based on the use of modified formulas for third-order aberration coefficients, this method enables one to decide if the individual members of the optical system can be simple lenses or if these should be more complex elements (cemented doublets, triplets, etc.). As a final result, one obtains the design parameters of the above-mentioned optical system (radii of curvature, optical materials, axial separations between individual elements). The analysis is performed for a thin-lens representation of the system. The transition to the thick-lens optical system then can be done by mathematical methods of numerical optimization using commercially available optical design software. The proposed method is shown on a practical example of calculation of parameters of such an optical system.

General method for replacing a thin lens with a thick lens with the same value of Seidel aberration coefficient of spherical aberration or coma.

The paper deals with the problem of replacing a thin lens with a thick lens having similar third-order aberration properties as the thin lens. The equations that make possible the calculation of parameters of the thick lens, which has the same value of focal length (or transverse magnification) and one of the Seidel aberration coefficients (either Seidel aberration coefficient of spherical aberration or Seidel aberration coefficient of coma) as the thin lens for given position of the entrance pupil and object, are derived. The application of the proposed method for calculation of parameters of the thick lens is shown in examples.

First-principles calculations of magnetic properties for analysis of magnetization processes in rare-earth permanent magnets.

It has been empirically known that the coercivity of rare-earth permanent magnets depends on the size and shape of fine particles of the main phase in the system. Also, recent experimental observations have suggested that the atomic-scale structures around the grain-boundaries of the fine particles play a crucial role to determine their switching fields. In this article, we review a theoretical attempt to describe the finite temperature magnetic properties and to evaluate the reduction of the switching fields of fine particles of several rare-earth permanent magnetic materials based on an atomistic spin model that is constructed using first-principles calculations. It is shown that, over a wide temperature range, the spin model gives a good description of the magnetization curves of rare-earth intermetallic compounds such as R 2Fe14B (R= Dy, Ho, Pr, Nd, Sm) and SmFe12. The atomistic spin model approach is also used to describe the local magnetic anisotropy around the surfaces of the fine particles, and predicts that the rare-earth ions may exhibit planar magnetic anisotropy when they are on the crystalline-structure surfaces of the particles. The dynamical simulation of the atomistic spin model and the corresponding micromagnetic simulation show that the planar surface magnetic anisotropy causes a reduction in the switching field of fine particles by approximately 20-30%, which may be relevant to the atomic-scale surface effects found in the experimental studies.

In Vitro and In Vivo Electrochemical Measurement of Reactive Oxygen Species After Treatment with Anticancer Drugs.

In vivo monitoring of reactive oxygen species (ROS) in tumors during treatment with anticancer therapy is important for understanding the mechanism of action and in the design of new anticancer drugs. In this work, a platinized nanoelectrode is placed into a single cell for detection of the ROS signal, and drug-induced ROS production is then recorded. The main advantages of this method are the short incubation time with the drug and its high sensitivity which allows the detection of low intracellular ROS concentrations. We have shown that our new method can measure the ROS response to chemotherapy in tumor-bearing mice in real-time. ROS levels were measured in vivo inside the tumor at different depths in response to doxorubicin. This work provides an effective new approach for the measurement of intracellular ROS by platinized nanoelectrodes.

Nanoscale Mapping Reveals Functional Differences in Ion Channels Populating the Membrane of Primary Cilia.

BACKGROUND/AIMS: The primary cilium is a nanoscale membrane protrusion believed to act as a mechano-chemical sensor in a range of different cell types. Disruptions in its structure and signalling have been linked to a number of medical conditions, referred to as ciliopathies, but remain poorly understood due to lack of techniques capable of investigating signal transduction in cilia at nanoscale. Here we set out to use latest advances in nanopipette technology to address the question of ion channel distribution along the structure of primary cilium. METHODS: We used glass nanopipettes and Scanning Ion Conductance Microscopy (SICM) to image 3D topography of intact primary cilia in inner medullary collecting duct (IMCD) cells with nanoscale resolution. The high-resolution topographical images were then used to navigate the nanopipette along the structure of each cilium and perform spatially resolved single-channel recordings under precisely controlled mechanical and chemical stimulation. RESULTS: We have successfully obtained first single-channel recordings at specific locations of intact primary cilia. Our experiments revealed significant differences between the populations of channels present at the ciliary base, tip and within extra-ciliary regions in terms of mean conductance and sensitivity to membrane displacement as small as 100 nm. Ion channels at the base of cilium, where mechanical strain is expected to be the highest, appeared particularly sensitive to the mechanical displacement. CONCLUSION: Our results suggest the distribution of ion channels in the membrane of primary cilia is non-homogeneous. The relationship between the location and function of ciliary ion channels could be key to understanding signal transduction in primary cilia.

Replacing a thin lens by a thick lens.

This paper deals with the problem of replacing a thin lens by a thick lens with approximately the same properties as the thin lens. Equations enabling the calculation of the parameters of a thick lens that has the same focal length and the same value for one of the Seidel aberration coefficients (either the Seidel aberration coefficient of spherical aberration or the Seidel aberration coefficient of coma) as the thin lens are derived. A comparison of the proposed method for calculating the parameters of the thick lens with existing methods is given in examples. Further, the problem of replacing a thick lens made of optical glass with a given refractive index by another thick lens with a different refractive index but with the same focal length and the same Seidel aberration coefficient of spherical aberration is investigated.

Kv1.1 channelopathy abolishes presynaptic spike width modulation by subthreshold somatic depolarization.

Although action potentials propagate along axons in an all-or-none manner, subthreshold membrane potential fluctuations at the soma affect neurotransmitter release from synaptic boutons. An important mechanism underlying analog-digital modulation is depolarization-mediated inactivation of presynaptic Kv1-family potassium channels, leading to action potential broadening and increased calcium influx. Previous studies have relied heavily on recordings from blebs formed after axon transection, which may exaggerate the passive propagation of somatic depolarization. We recorded instead from small boutons supplied by intact axons identified with scanning ion conductance microscopy in primary hippocampal cultures and asked how distinct potassium channels interact in determining the basal spike width and its modulation by subthreshold somatic depolarization. Pharmacological or genetic deletion of Kv1.1 broadened presynaptic spikes without preventing further prolongation by brief depolarizing somatic prepulses. A heterozygous mouse model of episodic ataxia type 1 harboring a dominant Kv1.1 mutation had a similar broadening effect on basal spike shape as deletion of Kv1.1; however, spike modulation by somatic prepulses was abolished. These results argue that the Kv1.1 subunit is not necessary for subthreshold modulation of spike width. However, a disease-associated mutant subunit prevents the interplay of analog and digital transmission, possibly by disrupting the normal stoichiometry of presynaptic potassium channels.

Effect of evaporative cooling and altitude on dairy cows milk efficiency in lowlands.

The objective of this current work was to determinate the effect of high temperatures on milk production of dairy cows in southern Slovakia in the year 2015. The hypotheses that milk production is influenced by the altitude and cooling were tested. Production data included 227,500 test-day records belonging to 34 Holstein breed herds situated in lowlands, 115 to 150 m above sea level (ASL) and kept in free-stall housing. Dairy farms were classified into groups based on cooling system. The first group of cows (19 herds) was cooled evaporative (foggers) and forced ventilation, and the second group (15 herds) was using cooled only forced ventilation (automatically controlled fans in housing and feeding areas). During the period from May to September, 36 summer and 22 tropical days were recorded, 37 days had a mean thermal humidity index value above 72.0, and on 34 days we recorded mean values above 78.0. The highest milk yields were recorded at the altitude 1 (115 m ASL) (9219.0 kg year-1; 10327.0 kg year-1) and the lowest at the altitude 2 (126 m ASL) (7598.7 kg year-1; 8470.21 kg year-1) (P < 0.001). Dairy cows cooled evaporative milked significantly more milk than cows cooled only with forced air flow (9650.4 kg vs. 8528.0 kg; P < 0.001). Fat and protein production differed also significantly (364.0 kg vs. 329.5 kg, P < 0.001; 312.2 kg vs. 279.7 kg, P < 0.001). It can be concluded that not only heat stress but also location farm above sea level can affect milk production. Evaporative cooling associated with increased air velocity is the appropriate protection against high temperatures.

Formation of Phases in Reactively Sintered TiAl3 Alloy.

This work highlights new results on the synthesis of the TiAl3 intermetallic phase using self-propagating high-temperature synthesis. This method is considered a promising sintering route for intermetallic compounds. It was found that the reactions proceed in two stages. Below the melting point of aluminum, the Ti2Al5 phase forms at 450 °C after long annealing times by a direct solid-state reaction between the aluminum and titanium, and is converted consequently to TiAl3. This is a completely new finding; until now, many authors have believed in the preferential formation of the TiAl3 phase. The second stage, the self-propagating strongly exothermic reaction, proceeds above the melting point of aluminum. It leads to the formation of the TiAl3 phase accompanied by Ti2Al5 and Ti3Al phases. The reaction mechanism was shown in the form of chemical equations, which were supported by calculating Gibbs energy. Reaction temperatures (Tonset, Tmaximum, and Toffset) were determined after induction heating thanks to recording by an optical pyrometer. This finding provides completely new opportunities for the determination of activation energy at heating rates, in which common calorimeters are not able to detect a response or even measure. Now, the whole procedure will become accessible.

The Effect of Simultaneous Si and Ti/Mo Alloying on High-Temperature Strength of Fe3Al-Based Iron Aluminides.

The effect of phase composition and morphology on high-temperature strength in the compression of Fe-Al-Si-based iron aluminides manufactured by casting was investigated. The structure and high-temperature strength in the compression of three alloys-Fe28Al5Si, Fe28Al5Si2Mo, and Fe28Al5Si2Ti-were studied. Long-term (at 800 °C for 100 h) annealing was performed for the achievement of structural stability. The phase composition and grain size of alloys were primarily described by means of scanning electron microscopy equipped with energy dispersive analysis and Electron Backscatter Diffraction (EBSD). The phase composition was verified by X-ray diffraction (XRD) analysis. The effect of Mo and Ti addition as well as the effect of long-term annealing on high-temperature yield stress in compression were investigated. Both additives-Mo and Ti-affected the yield stress values positively. Long-term annealing of Fe28Al5Si-X iron aluminide alloyed with Mo and Ti deteriorates yield stress values slightly due to grain coarsening.

Paraxial design of a four-component zoom lens with zero separation of principal planes and fixed position of an image focal point composed of members with constant focal length.

Our paper presents a detailed theoretical analysis of a four-component zoom lens with coincident principal planes, fixed position of the image focal point, and an approximately corrected Petzval sum. New formulas enabling the calculation of paraxial parameters of such optical systems are derived, and the practical application of these formulas is shown in an example. The position of the optical center of these optical systems does not depend on the object distance (for a given value of focal length of the zoom), and the position of the image focal point (with respect to the last element of the system) is fixed during the operation of the zoom. Such optical systems could be used in various measuring systems in photogrammetry, computer vision, triangulation sensors, fringe projection techniques, surveying, machine vision, etc.

Three-element optical system with a given fixed position of optical center.

Our paper presents a detailed theoretical analysis of the three-element optical system with zero separation of principal planes and approximately corrected Petzval sum, which has the required value of the axial position of its optical center for a given value of focal length and a given value of axial distance of the image focal point from the last element of the system. Formulas that make it possible to calculate paraxial parameters of such types of optical systems are derived, and the application of the derived formulas is presented in several examples. Such optical systems represent a subset of the new family of optical systems with the constant position of the optical center. The optical center of such an optical system is at a given required axial position, and this position does not change with the object distance, which could be advantageous for certain applications.

Solid-Phase Synthesis of Phosphorothioate/Phosphonothioate and Phosphoramidate/Phosphonamidate Oligonucleotides.

We have developed a robust solid-phase protocol which allowed the synthesis of chimeric oligonucleotides modified with phosphodiester and O-methylphosphonate linkages as well as their P-S and P-N variants. The novel O-methylphosphonate-derived modifications were obtained by oxidation, sulfurization, and amidation of the O-methyl-(H)-phosphinate internucleotide linkage introduced into the oligonucleotide chain by H-phosphonate chemistry using nucleoside-O-methyl-(H)-phosphinates as monomers. The H-phosphonate coupling followed by oxidation after each cycle enabled us to successfully combine H-phosphonate and phosphoramidite chemistries to synthesize diversely modified oligonucleotide strands.

Nanoscale Imaging of Primary Cilia with Scanning Ion Conductance Microscopy.

Primary cilia are hair-like sensory organelles whose dimensions and location vary with cell type and culture condition. Herein, we employed scanning ion conductance microscopy (SICM) to visualize the topography of primary cilia from different cell types. By combining SICM with fluorescence imaging, we successfully distinguished between surface cilia that project outward from the cell surface and subsurface cilia that are trapped below it. The nanoscale structure of the ciliary pocket, which cannot be easily identified using a confocal fluorescence microscope, was observed in SICM images. Furthermore, we developed a topographic reconstruction method using current-distance profiles to evaluate the relationship between set point and topographic image and found that a low set point is important for detecting the true topography of a primary cilium using hopping mode SICM.

Paraxial design of four-component zoom lens with fixed position of optical center composed of members with variable focal length.

The paper presents paraxial analysis of special types of zoom lenses, which are composed of four members with variable focal length. The position of optical center of these systems is required to be fixed for a given value of focal length (i.e., the position of the optical center does not depend on object distance for given value of focal length of the zoom). The formulas that enable the calculation of the optical powers of individual members of such a zoom lens are derived from--and the practical application of the derived formulas is demonstrated with--an example. Such optical systems represent a completely new family of zoom optical systems with applications in measuring systems in photogrammetry, computer vision, triangulation sensors, fringe projection techniques, surveying, machine vision, and so forth.

Paraxial design of an optical element with variable focal length and fixed position of principal planes.

In this article, we analyze the problem of the paraxial design of an active optical element with variable focal length, which maintains the positions of its principal planes fixed during the change of its optical power. Such optical elements are important in the process of design of complex optical systems (e.g., zoom systems), where the fixed position of principal planes during the change of optical power is essential for the design process. The proposed solution is based on the generalized membrane tunable-focus fluidic lens with several membrane surfaces.

Tuning the hybridization properties of modified oligonucleotides: from flexible to conformationally constrained phosphonate internucleotide linkages.

The concept of conformational restriction leading to the preorganization of modified strands has proven to be successful and has afforded nucleic acid analogues with many interesting properties suitable for various biochemical applications. We utilized this concept to prepare a set of constrained oligonucleotides derived from 1,4-dioxane and 1,3-dioxolane-locked nucleoside phosphonates and evaluated their hybridization affinities towards their complementary RNA strands. With an increase of ΔTm per modification up to +5.2 °C, the hybridization experiments revealed the (S)-2',3'-O-phosphonomethylidene internucleotide linkage as one of the most Tm-increasing modifications reported to date. Moreover, we introduced a novel prediction tool for the pre-selection of potentially interesting chemical modifications of oligonucleotides.

Determination of point of incidence for the case of reflection or refraction at spherical surface knowing two points lying on the ray.

The paper is focused on the problem of determination of the point of incidence of a light ray for the case of reflection or refraction at the spherical optical surface, assuming that two fixed points in space that the sought light ray should go through are given. The requirement is that one of these points lies on the incident ray and the other point on the reflected/refracted ray. Although at first glance it seems to be a simple problem, it will be shown that it has no simple analytical solution. The basic idea of the solution is given, and it is shown that the problem leads to a nonlinear equation in one variable. The roots of the resulting nonlinear equation can be found by numerical methods of mathematical optimization. The proposed methods were implemented in MATLAB, and the proper function of these algorithms was verified on several examples.