Acetylcholine receptor at neuromuscular junctions by EM autoradiography using mask analysis and linear sources

Several methods of analyzing EM autoradiograms are now available. Two such procedures, the grain density distribution (or histogram) method and the mask method use the resolution of the EM autoradiographic technique to generate grain distributions expected from postulated sources, and compare these with the observed grains in the autoradiograms. These two methods are here compared in the analysis of label on linear sources: the distribution of labeled acetylcholine receptor (AChR) down the postjunctional folds of lizard and frog neuromuscular junctions. The receptors were labeled with I-25-α-bungarotoxin and the autoradiograms coated with the high resolution Kodak emulsion 129–01. We found that both methods gave similar results in confirming that the bulk of the AChR is concentrated on the thickened region of the membrane at the top ∼2000 A of the junctional folds, and that there may be a gradient of receptor concentration down the folds. The grain density distribution method is simpler, but does not lend itself easily to quantifying the extent of deviation from simple models. Although computer graphics is not necessary for either method, its use allows the expected grains from linear sources to be generated quickly, making the mask analysis a feasible routine method for assigning the extent of label in different membrane regions.     

A methodological basis for SEM autoradiography: biosynthesis and radioligand binding

A method is described for scanning electron microscope (SEM) autoradiography whereby preservation of high resolution cell surface details is retained together with degelatination of the emulsion without gross loss or redistribution of silver grains. This method should provide a convenient medium-sized marker for SEM (using secondary, backscattered electron and X-ray imaging) topographic studies of biosynthesized molecules, and of cell surface receptors and antigens, using indirect or direct labelling procedures with radio-labelled ligands.     

Image spread in dry-mounted tritium autoradiographs

A comparison has been made of autoradiographs prepared by conventional dipping methods and those prepared from the same material using a dry-mounting method developed for the study of receptor labelling. Resolution was found to be inferior, and image spread greater, in the dry-mounted preparations. The extent to which resolution and image spread were affected depended upon the distance between specimen and emulsion.     

雌激素受体电镜定位的微波照射技术

将微波照射与电镜细胞化学技术相结合确定雌激素受体在细胞内的位置。在样品的固定、包埋、聚合及孵育过程中,使用改进的微波照射技术,既提高了样品的制备质量,又将样品制备时间缩短到原来的1/3。     

Labelled-replica techniques: post-shadow labelling of intramembrane particles in freeze-fracture replicas

Three methods are described for direct post-fracture, post-shadow labelling of individual classes of intramembrane particles (IMPs) in freeze-fracture replicas of biological membranes. The P-face IMPs corresponding to the acetylcholine receptor complexes (AChRs) of vertebrate neuroeffector junctions are identified by post-replication labelling with ferritin-antibody complexes and with neurotoxin-biotin-avidin-colloidal gold affinity ligands. (The freeze-etch nomenclature of Branton et al., 1975, is used in this report.) These post-shadow labelling techniques resemble conventional en bloc labelling techniques except that the labelling reagents must penetrate a thin but discontinuous layer of platinum superimposed on the molecules of interest. In the ‘sectioned labelled-replica technique’, the replicated and labelled tissues are stained, embedded in plastic and sectioned parallel to the replica-tissue interfaces. In the direct ‘labelled-replica techniques’, the replicated and labelled samples are freeze-dried or critical point dried, the labelled surfaces are stabilized by carbon coating, and the underlying tissues are dissolved, allowing the labelled-replicas to be examined as conventional freeze-fracture replicas. The unshadowed side of each AChR IMP is shown to retain sufficient biochemical information to permit both immunospecific and neurotoxin specific labelling despite formaldehyde fixation, freezing, fracturing, platinum shadowing, and thawing in aqueous media. A new mixed ferricyanide-osmium staining method reveals electron opaque structures spanning the membrane bilayer in the same size, number and distribution as the labelled IMPs. These experiments demonstrate the feasibility of identifying individual IMPs in freeze-fracture replicas and may allow the identification of specific membrane lesions in human disease.     

Immunohistochemical localization of angiotensin II receptor types 1 and 2 in the mesenteric artery from spontaneously hypertensive rats

Angiotensin II plays a crucial role in the control of blood pressure, acting at AT1 or AT2 receptors, and can act as a potent vasoconstrictor of the peripheral vasculature inducing hypertrophy, hyperplasia, or both, in resistance arteries. The aim of the present study was to investigate whether the pattern of distribution of angiotensin AT1 and AT2 receptors on mesenteric artery sections differs in spontaneously hypertensive rats (SHR) versus their respective controls (Wistar-Kyoto [WKY] rats). Immunohistochemistry using anti-AT1 or anti-AT2 antibodies was performed on perfused-fixed/paraffin-embedded mesenteric arteries from SHR and WKY rats. 3,3'-Diaminobenzidine tetrahydrochloride (DAB; activated by hydrogen peroxide) staining revealed distinct AT1 and AT2 labeling of all artery layers (adventitia, media and intima) from WKY rats, whereas in SHR an abundant AT1 labeling was found in both intima and adventitia and a sparser labeling in the media. There was a vast reduction of AT2 labeling throughout all layers. These results suggest a crucial role for AT2 receptors in the pathogenesis of hypertension.     

新冠病毒蛋白及其细胞受体ACE2翻译后修饰的质谱分析

新冠肺炎(COVID-19)在全世界范围内造成了巨大的健康危机和不可估量的损失.新出现的变种病毒株表明,新冠病毒(SARS-CoV-2)可能会像流感病毒一样在人类社会中继续流行,成为一种长久的健康威胁.控制新冠病毒的传染和开发有效的治疗方法迫在眉睫.因此,找到合适的生物标志物以表明病理和生理状态是当务之急.蛋白质是生命...     

Vibration and noise in brake systems of vehicles. Part 2: Theoretical investigation techniques

The paper reviews theoretical research into vibroacoustic phenomena in transport brake systems. The mechanisms of occurrence of different types of noise and vibration in brakes are described. Analytical, numerical, and experiment-calculated methods for studying brake noise are discussed and their disadvantages from the point of view of obtaining adequate calculation models are analyzed. The results of prediction of the vibroacoustic activity of friction units based on calculation methods are considered.     

Anterograde tracing in the brain using autoradiography and HRP-histochemistry. A comparison at the ultrastructural level

The present paper decribes the use of ³H-leucine and WGA-HRP as anterograde tracers in the brain at the ultrastructural level. In addition the different aspects of the two tracing techniques are compared, including the analysis of the results, the reliability and relative efficiency.     

Simultaneously identifying S-phase labelled cells and immunogold-labelling of vinculin in focal adhesions

A new combination of autoradiography and immunolabelling techniques is presented that allows the simultaneous identification of both S‐phase cells and their focal adhesions using scanning electron microscopy. The technique allows both labels to be discerned visually by their unique shapes and location within and on the cell. S‐phase cells were radio‐labelled with a pulse of tritiated thymidine, selectively incorporated into synthesizing DNA. The cells were then immunogold‐labelled for the focal adhesion protein, vinculin, prepared for autoradiography, and embedded in resin. The resin was then polymerized before removing the substrate, to expose the embedded cell undersurface. Electron‐energy ‘sectioning’ of the sample by varying the accelerating voltage of the electron beam allowed separate S‐phase cell identification in one electron‐energy ‘section’ and visualization of immunogold label in another ‘section’, within the same cell. As a result of applying this technique it was possible to positively identify S‐phase cells and immunogold‐labelled focal adhesions on the same cell simultaneously, which could be used to quantify focal adhesion sites on different substrates.     

对城市暴雨洪水利用有关问题的探讨

解决城市暴雨问题,过去一味强调疏与排,这样做的结果是大量雨水从城市排水系统流走了.雨洪过后,城市依然喊渴.现在人们逐渐发现,城市其实非常需要雨水,城市暴雨洪水完全可以收集和利用,这是解决城市排水和缺水问题的有效途径.     

Recent advancements in glycoproteomic studies: Glycopeptide enrichment and derivatization,characterization of glycosylation in SARS CoV2, and interacting glycoproteins

Proteomics studies allow for the determination of the identity, amount, and interactions of proteins under specific conditions that allow the biological state of an organism to ultimately change. These conditions can be either beneficial or detrimental. Diseases are due to detrimental changes caused by either protein overexpression or underexpression caused by as a result of a mutation or posttranslational modifications (PTM), among other factors. Identification of disease biomarkers through proteomics can be potentially used as clinical information for diagnostics. Common biomarkers to look for include PTM. For example, aberrant glycosylation of proteins is a common marker and will be a focus of interest in this review. A common way to analyze glycoproteins is by glycoproteomics involving mass spectrometry. Due to factors such as micro- and macroheterogeneity which result in a lower abundance of each version of a glycoprotein, it is difficult to obtain meaningful results unless rigorous sample preparation procedures are in place. Microheterogeneity represents the diversity of glycans at a single site, whereas macroheterogeneity depicts glycosylation levels at each site of a protein. Enrichment and derivatization of glycopeptides help to overcome these limitations. Over the time range of 2016 to 2020, several methods have been proposed in the literature and have contributed to drastically improve the outcome of glycosylation analysis, as presented in the sampling surveyed in this review. As a current topic in 2020, glycoproteins carried by pathogens can also cause disease and this is seen with SARS CoV2, causing the COVID-19 pandemic. This review will discuss glycoproteomic studies of the spike glycoprotein and interacting proteins such as the ACE2 receptor.     

Ovarian hormones and fasting differentially regulate pituitary receptors for estrogen and gonadotropin‐releasing hormone in rabbit female

To investigate the mechanisms by which caloric restriction affects reproductive function in female rabbits, we measured, in animals intact or ovariectomized (OVX) estrogen‐primed and fed ad libitum or fasted for 48 h, the adenohypophysial expression of estrogen receptor‐alpha (ESR1) and gonadotropin releasing hormone receptor (GnRHR) and the dynamic secretion of LH following GnRH stimulation. Fasting increased the number of GnRHR‐immunoreactive (‐IR) cells in intact animals, whereas reduced the density of ESR1‐IR cells in OVX rabbits. Estrogen priming decreased the number of ESR1‐IR cells in fasted and OVX animals. Ovariectomy increased the number of ESR1‐IR cells in fed rabbits, but caused an opposite effect in both fed and fasted animals treated with estrogen. Fasting down regulated the mRNA levels for ESR1 and GnRHR. Estrogen‐priming reduced the abundance for ESR1 mRNA in both fed and fasted rabbits, and that for GnRHR in fasted rabbits. Ovariectomy halved ESR1 mRNA levels independently of treatment and feeding condition, whereas increased (P < 001) that for GnRHR in estrogen‐primed rabbits. In all rabbits, an LH surge occurred 30 min after GnRH injection but the lowest levels were found in intact fasted rabbits and the highest in fasted, estrogen‐primed animals. The LH profile was similar in intact and OVX rabbits and neither fasting nor estrogen priming modified it. In conclusion, fasting differentially modifies the ESR1 and GnRHR expression in the pituitary, depending on the presence of gonadal hormones, indicating complex interactions between metabolic signals and ovarian steroids. Microsc. Res. Tech. 77:201–210, 2014. © 2013 Wiley Periodicals, Inc.     

Prouhet-Thue-Morse序列和原子函数在物理和技术中的应用(英文)

本文系统地阐述了信号与图像处理中与原子函数理论和Prouhet-Thue-Morse序列相关的大量研究成果。     

实物逆向工程中的关键技术及其最新发展

逆向工程是将实物转变为CAD模型相关的数字化技术、几何模型重构技术和产品制造技术的总称。在各工业领域有广泛的应用。总结了逆向工程中的数据获取、数据预处理、曲面重建等主要技术,分析了一些比较典型的方法,并就其在国内、外最新的研究进展进行了介绍。     

Receptor binding occurrence and plasma levels of natriuretic peptides in response to sympathectomy

In the present investigation the relationship between the sympathetic nervous system and the endocardial levels of receptor binding sites for natriuretic peptides and the plasma content of atrial natriuretic peptide were analyzed in rats. In order to destruct the cardiac sympathetic nerve terminals, chemical sympathectomy with 6-hydroxydopamine was made in parallel with intravenous measurements of blood pressure and heart frequency. By use of immunohistochemical and enzyme-linked-immunosorbent techniques the expression of tyrosine hydroxylase-positive sympathetic nerve terminals and plasma levels of pro-atrial natriuretic peptide were determined, respectively. The occurrence of receptor binding sites for natriuretic peptides was examined by in vitro receptor autoradiography. In contrast to the marked occurrence of natriuretic peptide receptor binding sites seen in the ventricular endocardium of control rats, the sympathectomized rats exhibited a decreased number of binding sites for natriuretic peptides in the endocardium of both the right and left chambers. Interestingly, this was found in parallel with a significant decrease of systolic and diastolic blood pressure and increased plasma levels of pro-atrial natriuretic peptide in the treated group of rats. These findings, together with those in previous studies, give support to an idea that one part of the blood pressure-decreasing effects, seen in patients treated with beta-adrenergic blockade, might be through a reduction of the natriuretic clearance receptor C, then giving rise to increased levels of atrial natriuretic peptide.     

Electromagnetic interference modeling and suppression techniques in variable-frequency drive systems

Electromagnetic interference (EMI) causes electromechanical damage to the motors and degrades the reliability of variable-frequency drive (VFD) systems. Unlike fundamental frequency components in motor drive systems, high-frequency EMI noise, coupled with the parasitic parameters of the trough system, are difficult to analyze and reduce. In this article, EMI modeling techniques for different function units in a VFD system, including induction motors, motor bearings, and rectifier-inverters, are reviewed and evaluated in terms of applied frequency range, model parameterization, and model accuracy. The EMI models for the motors are categorized based on modeling techniques and model topologies. Motor bearing and shaft models are also reviewed, and techniques that are used to eliminate bearing current are evaluated. Modeling techniques for conventional rectifier-inverter systems are also summarized. EMI noise suppression techniques, including passive filter, Wheatstone bridge balance, active filter, and optimized modulation, are reviewed and compared based on the VFD system models.     

激光辐照制备银纳米基底及其用于葡萄糖检测

通过激光辐照的方法制备银纳米条纹表面增强拉曼散射基底,该基底具有制备方法简单、均匀性好、重复性强的优点。用巯基吡啶(4-MPY)作为探针分子,葡萄糖和葡萄糖氧化酶产生的过氧化氢会改变银纳米条纹的形貌,进而影响探针分子的表面增强拉曼光谱强度,从而实现葡萄糖的间接检测。得到的探针分子的表面增强拉曼光谱的强度与葡萄糖溶液的浓度具有较好的线性关系。实验结果表明,该基底在葡萄糖的检测及定量分析方面具有良好的应用前景。     

Flow measurement of CO2 in a binary gaseous mixture using an averaging Pitot Tube and Coriolis mass flowmeters

To combat the growing emissions of CO₂ from industrial processes, Carbon Capture and Storage (CCS) and Carbon Capture and Utilization technologies (CCU) have been accepted worldwide to address these pressing concerns. So as to efficiently manage material and financial losses across the entire stream, accurate accounting and monitoring through fiscal metering of CO₂ in CCS transportation pipelines are core and required features for the CCS technologies. Moreover, these technical requirements are part of the legal compliance schemes and guidelines from various regulatory bodies. The CO₂ transportation pipelines will likely have multiple inputs from different capture plants, each with varying composition of CO₂ and thus introducing impurities into the CO₂ stream. The presence of other ordinary or hydrocarbon gases in the CO₂ gas stream could affect the functionality of metering instruments by introducing additional errors, particularly in the case of volumetric flowmeters. In this study, volumetric and direct mass measurement methods for the flow measurement of CO₂ mixtures using two totally different metering principles are experimentally evaluated. An Averaging Pitot Tube with Flow Conditioning Wing (APT-FCW) and Coriolis mass flowmeters (CMF) are used to assess the flow metering of CO₂ in a binary gaseous mixture. Different gases (nitrogen, air, oxygen, argon and propane) are diluted as contaminants into the pure CO₂ gas flow for various mass fractions to produce an adulterated mixture of the CO₂ gas. Comparative analysis of the measurement results under these flow conditions relative to that of pure CO₂ gas show that the measurement error of the APT-FCW sensor increases with the mass fraction of the diluent component, and gases with density closer to that of CO₂ have a much lesser effect on the performance of the APT-FCW flow sensor for smaller mass fractions. The CMF proved to be very reliable in the gas combination processes and as a reference meter for the APT-FCW sensor. Further analytical observations are discussed in detail.