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[Display omitted] • SUMOylation modifies AKT1 activity but its role in pluripotency remains understudied. • SUMOylation of AKT1 does not affect its targeting to the cell membrane. • SUMOylation and the E17K mutation affect AKT1 nucleus/cytoplasm partition. • E17K mutation promotes a more homogeneous SUMOylation-dependent AKT1 distribution. • AKT1 SUMOylation and the E17K mutation impact on Nanog-chromatin interactions. AKT/PKB is a kinase involved in the regulation of a plethora of cell processes. Particularly, in embryonic stem cells (ESCs), AKT is crucial for the maintenance of pluripotency. Although the activation of this kinase relies on its recruitment to the cellular membrane and subsequent phosphorylation, multiple other post-translational modifications (PTMs), including SUMOylation, fine-tune its activity and target specificity. Since this PTM can also modify the localization and availability of different proteins, in this work we explored if SUMOylation impacts on the subcellular compartmentalization and distribution of AKT1 in ESCs. We found that this PTM does not affect AKT1 membrane recruitment, but it modifies the AKT1 nucleus/cytoplasm distribution, increasing its nuclear presence. Additionally, within this compartment, we found that AKT1 SUMOylation also impacts on the chromatin-binding dynamics of NANOG, a central pluripotency transcription factor. Remarkably, the oncogenic E17K AKT1 mutant produces major changes in all these parameters increasing the binding of NANOG to its targets, also in a SUMOylation dependent manner. These findings demonstrate that SUMOylation modulates AKT1 subcellular distribution, thus adding an extra layer of regulation of its function, possibly by affecting the specificity and interaction with its downstream targets. [ABSTRACT FROM AUTHOR]

This paper developed a Finite Capacity Scheduling (FCS) system for make-pack production based on a real case of an adhesive factory. The FCS determines production quantity of each machine to conform with resource capacities and due date of customer orders while minimizes related total cost. The total cost includes total production, inventory, and cleaning cost. A Mixed Integer Linear Programming (MILP) model is formulated and solved by LINGO software. The computational time is very long since the model has a lot of integer variables. Thus, the model is solved for a reasonable time and the best but not optimal solution is reported with the lower bound. This paper tries fixed horizon and rolling horizon scheduling methods. The fixed horizon plans for an entire horizon of 30 days. The rolling horizon plans for a sub-horizon of 10, 15, and 17 days. An overlapping of sub-horizons is applied to reduce end-of-horizon effect. Three scenarios (high, normal, and low) of demands are considered. The fixed horizon method is applied first to all scenarios of demand. If the best solution is far away from the lower bound, the rolling horizon method is applied. The results indicated that the rolling horizon method may significantly reduce the total cost with the same computational time. Moreover, the rolling horizon method is more applicable for a dynamic situation where customers frequently change orders. The proposed MILP model can generate reasonable solutions and they are useful for scheduling decision of make-pack production.

With a growth in online course offering in recent years, there is much research focusing on student performance and student learning outcomes. However, research focusing on characteristics and achievement in Family and Consumer Sciences (FCS) higher education courses was not found. The purpose of this study was to compare student characteristics and achievements in online and oncampus FCS courses. The original study collected data from students enrolled in either the online or face-to-face section of the same junior-level course so variables were consistent. The study was later extended to another university using the same research set-up, but with a freshman-level course. This small, limited study provides preliminary insights that cannot be generalized, yet allow FCS faculty to distinguish student characteristics and achievements associated with on-campus and online courses. While student characteristics varied slightly, student achievement for online and on-campus students were similar. Additional studies are needed to provide more in-depth comparisons of these delivery systems. Since few FCS studies have reported this topic, the findings from this study provide baseline data for designing more in-depth comparisons of students.

To elucidate the molecular details of the activation-associated clustering of epidermal growth factor receptors (EGFRs), the time course of the mobility and aggregation states of eGFP tagged EGFR in the membranes of Chinese hamster ovary (CHO) cells was assessed by in situ mobility assays. Fluorescence correlation spectroscopy (FCS) was used to probe molecular movements of small ensembles of molecules over short distances and time scales, and to report on the state of aggregation. The diffusion of larger ensembles of molecules over longer distances (and time scales) was investigated by fluorescence recovery after photobleaching (FRAP). Autocorrelation functions could be best fitted by a two-component diffusion model corrected for triplet formation and blinking. The slow, 100−1000 ms component was attributed to membrane localized receptors moving with free Brownian diffusion, whereas the fast, ms component was assigned to cytosolic receptors or their fragments. Upon stimulation with 50 nM EGF, a significant decrease from 0.11 to 0.07 μm2/s in the diffusion coefficient of membrane-localized receptors was observed, followed by recovery to the original value in ~20 min. In contrast, the apparent brightness of diffusing species remained the same. Stripe FRAP experiments yielded a decrease in long-range molecular mobility directly after stimulation, evidenced by an increase in the recovery time of the slow component from 13 to 21.9 s. Our observations are best explained by the transient attachment of ligand-bound EGFRs to immobile or slowly moving structures such as the cytoskeleton or large, previously photobleached receptor aggregates.

A new block copolymer with a temperature-responsive block and a cationic block was prepared by reversible addition-fragmentation chain transfer (RAFT) polymerization, with good control of its size and composition. The first block is composed by di(ethylene glycol)methyl ethermethacrylate (DEGMA) and oligo(ethylene glycol) methyl ether methacrylate (OEGMA), with the ratio DEGMA/OEGMA being used to choose the volume phase transition temperature of the polymer in water, tunable from ca. 25 to above 90 °C. The second block, of trimethyl-2-methacroyloxyethylammonium chloride (TMEC), is positively charged at physiological pH values and is used for DNA binding. The coacervate complexes between the block copolymer and a model single strand DNA are characterized by fluorescence correlation spectroscopy and fluorescence spectroscopy. The new materials offer good prospects for biomedical application, for example in controlled gene delivery. [ABSTRACT FROM AUTHOR]

Background: Familial chylomicronemia syndrome (FCS) is a rare, inherited lipid disorder characterized by high levels of plasma triglycerides and chylomicrons, which may cause life-threatening acute pancreatitis. Currently no FDA-approved treatment exists. Management is low-fat diet (<20g fat/day), which is difficult to maintain. With the restricted diet, triglycerides may remain elevated. We conducted discussions with patients and caregivers to better understand the burden of FCS from their perspectives.Methods: A panel of FCS patients and caregivers was assembled to discuss and assess the clinical and psychosocial burden of FCS.Results: Ten adults with FCS (median age 48 yr) and their spouses/caregivers were asked specific questions about their experiences living with FCS. Patients with FCS stated their symptoms were abdominal pain, nausea, diarrhea, constipation, bloating, and fatigue. Patients reported a median of 34 episodes of acute pancreatitis over their lifetimes; half of these led to hospitalizations, each with an average stay of 6.5 days. The psychosocial burden of FCS was primarily associated with the restricted diet, anxiety and stress of FCS.Conclusions: Living with FCS imposes a significant clinical and psychosocial burden on patients and caregivers, who reported reduced quality of life, limited employment opportunities, socialization and increased burden on family. [ABSTRACT FROM AUTHOR]

The Electron Cyclotron Upper Launcher is an eight beamline ITER antenna whose main goal is to drive current locally, inside magnetic islands that may form on the q = 3/2 or 2 rational magnetic flux surfaces, in order to stabilize neoclassical tearing modes (NTMs). The primary vacuum boundary at the port plug extends into the port cell region through the ex-vessel mm-wave waveguide components, defining the so-called First Confinement System (FCS). Thermal expansion, seismic events and plasma disruption events result in displacements of the vacuum vessel, relative to the tokamak building, that are transferred to the FCS at its interfaces with the port plug. In absence of suitable inline waveguide bellows, the adaptation to such imposed displacements is provided by bending compliance of the straight waveguide sections. An analysis methodology for the ex-vessel components was developed which covers a preliminary set of applicable load combinations throughout the system life-cycle. This methodology describes the FCS numerical model developed to assess the system against independent events as well as the load combination strategy followed to combine the results. Some simplifications like the simulation of the Vertical Displacement Events (VDEs) by a static approach are considered in order to develop this methodology. The analysis of the results shows that the miter bends and the pieces of waveguides rigidly attached to the port cell ceiling are the most demanded components for most of the considered load combinations. [ABSTRACT FROM AUTHOR]

Understanding the colloidal properties of extracellular vesicles (EVs) is key to advance fundamental knowledge in this field and to develop effective EV-based diagnostics, therapeutics and devices. Determination of size distribution and of colloidal stability of purified EVs resuspended in buffered media is a complex and challenging issue – because of the wide range of EV diameters (from 30 to 2000 nm), concentrations of interest and membrane properties, and the possible presence of co-isolated contaminants with similar size and densities, such as protein aggregates and fat globules – which is still waiting to be fully addressed. We report here a fully detailed protocol for accurate and robust determination of the size distribution and stability of EV samples which leverages a dedicated combination of Fluorescence Correlation Spectroscopy (FCS) and Dynamic Light Scattering (DLS). The theoretical background, critical experimental steps and data analysis procedures are thoroughly presented and finally illustrated through the representative case study of EV formulations obtained from culture media of B16 melanoma cells, a murine tumor cell line used as a model for human skin cancers. [ABSTRACT FROM AUTHOR]

SAGA and ATAC are two distinct chromatin modifying co-activator complexes with distinct enzymatic activities involved in RNA polymerase II (Pol II) transcription regulation. To investigate the mobility of co-activator complexes and general transcription factors in live-cell nuclei, we performed imaging experiments based on photobleaching. SAGA and ATAC, but also two general transcription factors ( TFIID and TFIIB), were highly dynamic, exhibiting mainly transient associations with chromatin, contrary to Pol II, which formed more stable chromatin interactions. Fluorescence correlation spectroscopy analyses revealed that the mobile pool of the two co-activators, as well as that of TFIID and TFIIB, can be subdivided into 'fast' (free) and 'slow' (chromatin-interacting) populations. Inhibiting transcription elongation decreased H3K4 trimethylation and reduced the 'slow' population of SAGA, ATAC, TFIIB and TFIID. In addition, inhibiting histone H3K4 trimethylation also reduced the 'slow' populations of SAGA and ATAC. Thus, our results demonstrate that in the nuclei of live cells the equilibrium between fast and slow population of SAGA or ATAC complexes is regulated by active transcription via changes in the abundance of H3K4me3 on chromatin. [ABSTRACT FROM AUTHOR]

Mitochondria are cellular organelles with multifaceted tasks and thus composed of different sub-compartments. The inner mitochondrial membrane especially has a complex nano-architecture with cristae protruding into the matrix. Related to their function, the localization of mitochondrial membrane proteins is more or less restricted to specific sub-compartments. In contrast, it can be assumed that membrane proteins per se diffuse unimpeded through continuous membranes. Fluorescence recovery after photobleaching is a versatile technology used in mobility analyses to determine the mobile fraction of proteins, but it cannot provide data on subpopulations or on confined diffusion behavior. Fluorescence correlation spectroscopy is used to analyze single molecule diffusion, but no trajectory maps are obtained. Single particle tracking (SPT) technologies in live cells, such as tracking and localization microscopy (TALM), do provide nanotopic localization and mobility maps of mitochondrial proteins in situ. Molecules can be localized with a precision of between 10 and 20 nm, and single trajectories can be recorded and analyzed; this is sufficient to reveal significant differences in the spatio-temporal behavior of diverse mitochondrial proteins. Here, we compare diffusion coefficients obtained by these different technologies and discuss trajectory maps of diverse mitochondrial membrane proteins obtained by SPT/TALM. We show that membrane proteins in the outer membrane generally display unhindered diffusion, while the mobility of inner membrane proteins is restricted by the inner membrane architecture, resulting in significantly lower diffusion coefficients. Moreover, tracking analysis could discern proteins in the inner boundary membrane from proteins preferentially diffusing in cristae membranes, two sub-compartments of the inner mitochondrial membrane. Thus, by evaluating trajectory maps it is possible to assign proteins to different sub-compartments of the same membrane. [ABSTRACT FROM AUTHOR]

Nonspecific interactions between lipids and fluorophores can alter the outcomes of single-molecule spectroscopy of membrane proteins in live cells, liposomes or lipid nanodiscs and of cytosolic proteins encapsulated in liposomes or tethered to supported lipid bilayers. To gain insight into these effects, we examined interactions between 9 dyes that are commonly used as labels for single-molecule fluorescence (SMF) and 6 standard lipids including cationic, zwitterionic and anionic types. The diffusion coefficients of dyes in the absence and presence of set amounts of lipid vesicles were measured by fluorescence correlation spectroscopy (FCS). The partition coefficients and the free energies of partitioning for different fluorophore-lipid pairs were obtained by global fitting of the titration FCS curves. Lipids with different charges, head groups and degrees of chain saturation were investigated, and interactions with dyes are discussed in terms of hydrophobic, electrostatic and steric contributions. Fluorescence imaging of individual fluorophores adsorbed on supported lipid bilayers provides visualization and additional quantification of the strength of dye-lipid interaction in the context of single-molecule measurements. By dissecting fluorophore-lipid interactions, our study provides new insights into setting up single-molecule fluorescence spectroscopy experiments with minimal interference from interactions between fluorescent labels and lipids in the environment. [ABSTRACT FROM AUTHOR]

In this paper, new fully floating memristor emulator circuit that consumes ultra-low energy is presented. This proposed circuit is simple because of the fact that it doesn't contain any multiplication or various block circuits to obtain nonlinear characteristics of memristor. Transistors are operated in subthreshold region to obtain the non-linear behavior. Floating Current Source (FCS) is used in memristor emulator design to decrease the energy consumption of emulator. Associative learning is a type of the learning mechanisms and designed memristor are used in classical associative learning circuit successfully. All results are compatible with both memristor characteristics and learning mechanisms of circuit. [ABSTRACT FROM AUTHOR]

Translational diffusion properties of single-stranded DNA (ssDNA) and G-quadruplexes were studied to determine the persistence length and cooperativity of G-quadruplex formation using FCS combined with HYDRO in which wormlike chain (WLC)-based Monte Carlo simulation are implemented. The presence of a guanine instead of a thymine shortened the contour length of nucleic acids and increased the vulnerability to ion screening. For cooperativity estimation, the telomeric sequence HT was assumed to undergo 27 intermediate states, which can be classified as ssDNA, single-G-quadruplex, double-G-quadruplex, and three consecutive G-quadruplexes. Each state type was modeled using a series of beads and appropriate bond lengths, which were obtained from the WLC model. Using the HYDRO program, we calculated diffusion times for each species, and these were used to calculate simulated HT diffusion times for mixtures of species in arbitrary KCl concentrations. By comparison between simulated and experimental diffusion properties, we obtained a positive cooperativity of C = 200 from FCS combined with HYDRO. [ABSTRACT FROM AUTHOR]

Protein is essential to human health, but its digestion kinetics in varied structures are not yet well understood. We previously found different kinetics of protein hydrolysis in solution and in gels, and we hypothesized that the difference stemmed from the steric hindrance of gel structure to the diffusion of pepsin and its hydrolysates. To better understand the pepsin diffusivity in food matrices and its effect on digestion, we determined the diffusivity of pepsin in water and in whey protein isolate (WPI) gels by fluorescence correlation spectroscopy (FCS). We estimated the pepsin concentration gradient during digestion based on the determined diffusivity, which showed that the pepsin is constrained within a thin layer from the gel surface. Gel composition analysis confirmed this constraint: peptides as protein fragments were observed only in the first 2 mm of the WPI gels after 6 h of in vitro gastric digestion. Scanning electron microscopy indicated that pepsin loosened the microstructure of whey protein gel surfaces, which may accelerate pepsin diffusion and consequently gel surface disintegration. We conclude that the mode of whey protein gel digestion is determined by the summed effect of diffusion limitation, hydrolysis rate and microstructure transformation. [ABSTRACT FROM AUTHOR]

This study presents an online multiparameter estimation scheme for interior permanent magnet motor drives that exploits the switching ripple of finite control set (FCS) model predictive control (MPC). The combinations consist of two, three, and four parameters are analysed for observability at different operating states. Most of the combinations are rank deficient without persistent excitation (PE) of the system, e.g. by signal injection. This study shows that high frequency current ripples by MPC with FCS are sufficient to create PE in the system. This study also analyses parameter coupling in estimation that results in wrong convergence and propose a decoupling technique. The observability conditions for all the combinations are experimentally validated. Finally, a full parameter estimation along with the decoupling technique is tested at different operating conditions. [ABSTRACT FROM AUTHOR]

Background: Familial Chylomicronemia Syndrome (FCS) is a rare genetic disorder that is caused by a decrease or an absence of lipoprotein lipase activity. FCS is characterized by marked accumulation of chylomicrons and extreme hypertriglyceridemia, which have major effects on both physical and mental health. To date, there have been no systematic efforts to characterize the impact of chylomicronemia on FCS patients' lives. In particular, the impact of FCS on the burden of illness (BoI) and quality of life (QoL) has not been fully described in the literature.Methods: IN-FOCUS was a comprehensive web-based research survey of patients with FCS focused on capturing the BoI and impact on QoL associated with FCS. Sixty patients from the US diagnosed with FCS participated. Patients described multiple symptoms spanning across physical, emotional and cognitive domains.Results: Patients on average cycled through 5 physicians of varying specialty before being diagnosed with FCS, reflecting a lengthy journey to diagnosis Nearly all respondents indicated that FCS had a major impact on BoI and QoL and significantly influenced their career choice and employment status, and caused significant work loss due to their disease.Conclusion: FCS imparts a considerable burden across multiple domains with reported impairment on activities of daily living and QoL. [ABSTRACT FROM AUTHOR]

Education and problems related to education are greatly influenced by economic, political and social factors. This cross-cultural comparison of the educational systems in the United States and South Korea shed light on how these factors influence the value placed on Family and Consumer Sciences/Home Economics curricular offerings. This study is a qualitative comparison of the structure and offerings of Family and Consumer Sciences secondary programs to those in South Korea. The information was gathered through interviews and overning policies and educational mandate documents. The educational systems differ in many ways; however, secondary programs in family and consumer sciences/Home Economics have a long history in both countries. In the United States, the courses and opportunities vary in state requirements. There are two types of high schools in South Korea, general and vocational. Admission to high school is dependent on the results of competitive entrance examinations. Home economics is one of the optional programs as is Art, Music, Physical Education, and Technology. The subject of clothing and textiles receives major emphasis in South Korea. In the United States, due to the emphasis on societal needs along with budgetary cuts, clothing and textile programs in secondary schools are often one of the first to be cut. In South Korea, consumer education is entirely neglected throughout the four years of homemaking education; contrary, there is currently a push in the United States to mandate personal finance courses in the public schools, the catalyst being the huge consumer debt. A follow-up empirical study is recommended. [ABSTRACT FROM AUTHOR]

The EU Habitats Directive is a key biodiversity conservation instrument. It contains legal obligations for the 28 EU member states in order to safeguard a 'favourable conservation status' (FCS) for selected species and habitat types. The crucial FCS concept itself, however, remains subject to considerable confusion regarding its proper interpretation and operationalization, impairing the Directive's effective implementation. Diminishing this confusion is the purpose of this review. It focuses specifically on large carnivores-wolf ( Canis lupus), brown bear ( Ursus arctos), Eurasian lynx ( Lynx lynx) and wolverine ( Gulo gulo). These pose particular challenges, given their low densities, transboundary populations, and human-wildlife conflict potential. Large carnivores are also the only species for which specific guidance has been adopted by the European Commission-and subsequently contested. Our methodology combines legal analysis with an understanding of the species' ecology and associated social, economic and cultural dimensions. We analyze the methods and processes through which EU law is interpreted, implemented, and enforced, by member states, European Commission, and EU Court of Justice-which is the ultimate authority regarding EU law interpretation. On that basis, we engage three particularly complex interpretation questions which are also of great practical significance: (1) the appropriate scale to achieve FCS (national or transboundary population level); (2) the respective roles of demographic, genetic and ecological factors in determining FCS; and (3) the use of extinction versus carrying capacity as benchmark. Regarding these questions, we identify approaches that are workable and effective, as well as likely to be endorsed by the EU Court. [ABSTRACT FROM AUTHOR]

The combination of nanostructured fluids (NSFs), such as micellar solutions or microemulsions and highly-retentive hydrogels, such as PVA-based cryogels, represents nowadays a promising and innovative approach in the field of conservation of cultural heritage, as it enables the highest control available during cleaning interventions. However, at present, little is known about the confinement of NSFs into PVA-based cryogels. In the present work, small-angle X-ray scattering (SAXS) and ultra-small-angle X-ray scattering (USAXS), combined with fluorescence correlation spectroscopy (FCS), were used to address this issue. Two significantly different NSFs were selected for this purpose, based on anionic or nonionic surfactants, and they were confined into four different gels: two single PVA and two PVA/PVA "twin-chain polymer networks" (TC-PNs). The analysis of the experimental results shows that surfactant nature is crucial in determining the interaction of NSFs with the gels' polymer network. Moreover it was demonstrated that loading NSFs on PVA-based cryogels does not significantly affect the short-range nanostructure of neither the NSFs nor the gels, preserving the cleaning performances of the combined system. [Display omitted] • Nanostructured fluids (NSFs) confined in cryogels represent a powerful cleaning tool. • USAXS and FCS were used to study the nanostructure of PVA cryogels/NSFs systems. • USAXS allowed the study of the PVA cryogels' nanostructure on a larger length scale. • NSFs induced minor changes in the gels' nanostructure, i.e. a slight increase in crystallites' size. • FCS clarified that surfactant's nature is key to determine NSF/gel interactions. [ABSTRACT FROM AUTHOR]

Unraveling the transport of drugs and nanocarriers in cerebrovascular networks is important for pharmacokinetic and hemodynamic studies but is challenging due to the complexity of sensing individual particles within the circulatory system of a live animal. Here, we demonstrate that a DNA-stabilized silver nanocluster (DNA-Ag 16 NC) that emits in the first near-infrared window upon two-photon excitation in the second NIR window can be used for multiphoton in vivo fluorescence correlation spectroscopy for the measurement of cerebral blood flow rates in live mice with high spatial and temporal resolution. To ensure bright and stable emission during in vivo experiments, we loaded DNA-Ag 16 NCs into liposomes, which served the dual purposes of concentrating the fluorescent label and protecting it from degradation. DNA-Ag 16 NC-loaded liposomes enabled the quantification of cerebral blood flow velocities within individual vessels of a living mouse.

This paper addresses the energy management strategy (EMS) for a fuel cell hybrid electric vehicle (FC‐HEV). The fuel cell system (FCS) is a multi‐physics system, and consequently, its energetic performances depend on the degradation and on the operating conditions. The maximum power (MP) and the maximum efficiency (ME) points of the FCS are unique but they move with operating condition variations. Thus, developing an extremum seeking process (ESP) for both MP and ME tracking is a challenging task. In the ESP, models are identified online by using an adaptive recursive least square (ARLS) method to seek a variation in the FCS performances. Then an optimisation algorithm is used on the updated model to find the MP and the ME points. The ESP is incorporated into a hysteresis power splitting control (HPSC). A MP mode or a ME mode can be set based on the energy storage level (battery pack). The effectiveness of the proposed MP‐ and ME‐ESP EMS is demonstrated by conducting experimental studies on two FCSs with different levels of degradation. It was demonstrated that the classical EMS based on maps are not valid when the operating parameters vary because of the level of degradation change. [ABSTRACT FROM AUTHOR]

Extracellular proteins devoid of signal peptides use nonclassical secretion mechanisms for their export. These mechanisms are independent of the endoplasmic reticulum and Golgi. Some nonclassically released proteins, particularly fibroblast growth factors (FGF) 1 and 2, are exported as a result of their direct translocation through the cell membrane. This process requires specific interactions of released proteins with membrane phospholipids. In this review written by a cell biologist, a structural biologist and two membrane engineers, we discuss the following subjects: (i) Phenomenon of nonclassical protein release and its biological significance; (ii) Composition of the FGF1 multiprotein release complex (MRC); (iii) The relationship between FGF1 export and acidic phospholipid externalization; (iv) Interactions of FGF1 MRC components with acidic phospholipids; (v) Methods to study the transmembrane translocation of proteins; (vi) Membrane models to study nonclassical protein release.

Formation of the three embryonic germ layers is a fundamental developmental process that initiates differentiation. How the zebrafish pluripotency factor Pou5f3 (homologous to mammalian Oct4) drives lineage commitment is unclear. Here, we introduce fluorescence lifetime imaging microscopy and fluorescence correlation spectroscopy to assess the formation of Pou5f3 complexes with other transcription factors in real-time in gastrulating zebrafish embryos. We show, at single-cell resolution in vivo, that Pou5f3 complexes with Nanog to pattern mesendoderm differentiation at the blastula stage. Later, during gastrulation, Sox32 restricts Pou5f3–Nanog complexes to the ventrolateral mesendoderm by binding Pou5f3 or Nanog in prospective dorsal endoderm. In the ventrolateral endoderm, the Elabela / Aplnr pathway limits Sox32 levels, allowing the formation of Pou5f3–Nanog complexes and the activation of downstream BMP signaling. This quantitative model shows that a balance in the spatiotemporal distribution of Pou5f3–Nanog complexes, modulated by Sox32, regulates mesendoderm specification along the dorsoventral axis.

This paper discusses recent advances in confocal laser scanning microscopy (CLSM) for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm) tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer), FLIM (Fluorescence Lifetime Imaging Microscopy), FCS (Fluorescence Correlation Spectroscopy) and FRAP (Fluorescence Recovery After Photobleaching) are introduced and their applicability for quantitative imaging of biomolecular (co-)localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.

Human Immunodeficiency Virus type-1 (HIV-1) acquires its lipid membrane from the plasma membrane of the infected cell from which it buds out. Previous studies have shown that the HIV-1 envelope is an environment of very low mobility, with the diffusion of incorporated proteins two orders of magnitude slower than in the plasma membrane. One of the reasons for this difference is thought to be the HIV-1 membrane composition that is characterised by a high degree of rigidity and lipid packing, which has, until now, been difficult to assess experimentally. To further refine the model of the molecular mobility on the HIV-1 surface, we herein investigated the relative importance of membrane composition and curvature in simplified model membrane systems, large unilamellar vesicles (LUVs) of different lipid compositions and sizes (0.1–1 µm), using super-resolution stimulated emission depletion (STED) microscopy-based fluorescence correlation spectroscopy (STED-FCS). Establishing an approach that is also applicable to measurements of molecule dynamics in virus-sized particles, we found, at least for the 0.1–1 µm sized vesicles, that the lipid composition and thus membrane rigidity, but not the curvature, play an important role in the decreased molecular mobility on the vesicles’ surface. This observation suggests that the composition of the envelope rather than the particle geometry contributes to the previously described low mobility of proteins on the HIV-1 surface. Our vesicle-based study thus provides further insight into the dynamic properties of the surface of individual HIV-1 particles, as well as paves the methodological way towards better characterisation of the properties and function of viral lipid envelopes in general.

In view of the serious problem of packet loss existed in transmitting audio and video streaming by use of wireless sensor network technology in coal mine underground,an Ethernet data unpacked/packed method based on FPGA was put forward,namely,an Ethernet data frame would be unpacked into several small packets for transmission at the sending end,while these small data packets would be repacked as an Ethernet data frame at the receiving end,and some processing to the lost or damaged data would be done.Finally,frame check sequence(FCS) of the received data frame will be(re-calculated.) The method makes use of the frame which losses part of the data effectively,improves utilization of effective data,and reduces impact on quality of the received audio and video streaming caused by the loss of audio and video stream data.

RNA double strand hybridization is a hallmark for gene expression regulation. In this function, single stranded regulatory RNA forms Watson-Crick base pairs with complementary messenger RNA. In the presented work the dissociation constants of complementary equally sized RNA single strands were measured at the single molecule level applying fluorescence correlation spectroscopy (FCS). Dissociation constants of 3.2 nM, 1.4 nM and 1.0 nM were determined for 26 bp, 41 bp and 54 bp dsRNA, respectively. The translational diffusion coefficients of RNA, measured at infinite dilution, could be accurately predicted applying the model D = 4.58 × 10−10 N−0.39 m2s−1. [Display omitted] • Hybridization of RNA single strands measured applying FCS. • Dissociation constants of antisense RNA and siRNA. • Prediction of diffusion coefficients of RNA by an empirical model. [ABSTRACT FROM AUTHOR]

D614G mutation plays a significant role in the transmissibility of SARS-CoV-2. Identification of other mutations related to D614G mutation within the Spike protein is pivotal as they might contribute to the pathogenicity of SARS-CoV-2. This study aims to analyze the mutation rate of furin cleavage site (FCS) region of Indonesian origin SARS-CoV-2 and to predict the effect of mutation against Spike priming efficiency by furin. A total of 375 sequences of Indonesian isolates obtained during the early pandemic were used for mutation analysis. Mutation analysis includes mutation pattern, variability, frequency of mutation, amino acid conservation, and mutation rate. The effect of mutation against Spike priming efficiency by furin protease from eight sequences with mutation in the FCS region was analyzed by protein-protein docking. We showed that mutations related to the G614 variant were increasing through time, in contrast to the D614 variant. The FCS region at the position 675-692 contained the most variable (66.67%) as well as the highest mutation frequency (85.92%) and has been observed to be the hotspot mutations linked to the D614G mutation. The D614G hotspot-FCS region (residue 600-700) had the highest amino acid change per site (20.8%) as well as the highest mutation rate as 1.34 × 10 -2 substitution per site per year (95% CI 1.79 × 10 -3 -2.74 × 10 -2 ), compared with other Spike protein regions. Mutations in the FCS region were the most common mutation found after the D614G mutation. These mutations were predicted to increase the Spike priming efficiency by furin. Thus, this study elucidates the importance of D614G mutation to other mutations located in the FCS region and their significance to Spike priming efficiency by furin., Supplementary Information: The online version contains supplementary material available at 10.1007/s13337-023-00827-w., Competing Interests: Conflict of interestThe authors declare no conflict of interest., (© The Author(s), under exclusive licence to Indian Virological Society 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)