We have recently developed surface oligopeptide knock-in for rapid target selection (SORTS), a novel method to isolate mammalian cells with gene modifications using FACS-sorting. It relies on CRISPR/Cas9-mediated targeted knock-in of a very short promoterless expression construct (250 bp) comprising a Flag or HA epitope embedded into the smallest GPI-protein CD52, and a polyA signal from the beta-globin. CD52 efficiently delivers the epitope to the cell surface, where it serves as a marker for selection, while polyA terminates transcription and silences target gene expression. Primarily, SORTS was developed to knock out genes encoding intracellular or secreted proteins, which cannot be used as markers for selection of live cells. Using in-frame modification of SORTS, we demonstrated the possibility of HIV-1 provirus inactivation via sorting of GPI-tag positive cells. In order to make the “cured” cells resistant to a subsequent HIV-1 infection, the epitope tag in the CD52 molecule was substituted by one of the fusion inhibitor peptides from the CHR-domain of gp41. We selected a series of cell-surface-expressed, GPI-anchored, C34-based peptides that confer a strong cellular resistance to HIV-1 infection mediated by NL4-3, JRFL, or ZM153 Env. These findings together with a monoclonal antibody raised against the C34 peptide provide an opportunity to generate and select HIV-resistant lymphocytes for a therapeutic goal. SORTS was also adapted to engineer transgenic HIV-1 effector Т cells and to study cell-to-cell transmission. To facilitate transgenesis, we developed a knock-in strategy to express GPI-tag from the intronic region of the human PPP1R12C gene (AAVS1 locus) and delivered FRT sites of recombination into both alleles. In summary, SORTS is a novel instrument to isolate rare cells with precise genomic modifications with broad applications, including HIV biology. This work was supported by the Russian Science Foundation (grant 18-14-00333) and the Russian Foundation for Basic Research (grants 18-29-07052, 18-04-01016). Full article

Tick-borne encephalitis virus (TBEV) is a pathogen that causes severe human neuroinfections in Europe and Asia for which there is currently no specific therapy. The adenosine analogue galidesivir (BCX4430), a broad-spectrum RNA virus inhibitor, has entered a phase 1 clinical safety and pharmacokinetics study in healthy subjects and is under clinical development for treatment of Ebola and yellow fever virus infections. Moreover, galidesivir also inhibits the reproduction of TBEV and numerous other medically important flaviviruses. Until now, studies of this antiviral agent have not yielded resistant viruses. In our study, we performed serial in vitro passaging of TBEV in the presence of increasing concentrations of galidesivir (up to 50 μM), which resulted in the generation of two drug-resistant TBEV mutants. The first TBEV mutant was characterized by a single amino acid change, E460D. The other carried two amino acid changes, E460D and Y453H. Both mutations mapped to the active site of the viral RNA-dependent RNA polymerase (RdRp). Galidesivir-resistant TBEV exhibited no cross-resistance to structurally different antiviral nucleoside analogues, such as 7-deaza-2′-C-methyladenosine, 2′-C-methyladenosine, and 4′-azido-aracytidine. Although the E460D substitution led to only a subtle decrease in viral fitness in cell culture, galidesivir-resistant TBEV was highly attenuated in vivo, with a 100% survival rate and no clinical signs observed in infected mice. Furthermore, no virus was detected in the sera, spleen, or brain of mice inoculated with the galidesivir-resistant TBEV. By contrast, infection with wild-type virus resulted in fatal infections for all animals. Our results contribute to understanding the molecular basis of galidesivir antiviral activity, flavivirus resistance to nucleoside inhibitors, and the potential contribution of viral RdRp to flavivirus neurovirulence. Full article

The Western Pacific Warm Pool (WP), with the highest sea surface temperature (SST) in the world, has strong impacts on the drought variations in Eurasia. However, since the little ice age (1250–1850, LIA), the co-climatic drought pattern due to WP warming in Eurasia remains unclear. This is a long-term warming background for the current warming period (CWP). In this paper, we use both instrumental data and 1625 tree-ring width records from Eurasia to investigate the drought patterns in both modern and historical periods. This study revealed two seesaw precipitation patterns, namely the Central Asia–Mongolia (CAMO) and Northern Europe–Southern Europe (NESE) patterns. When the Western Pacific Warm Pool sea surface temperature (WPSST) is high, precipitation increases in Central Asia and Northern Europe, and decreases in Mongolia and southern Europe. When the positive (negative) phase event of the El Niño–Southern Oscillation (ENSO) occurs, the WPSST is reduced (increased), and the decreases (increases) of precipitation in Central Asia and Northern Europe and the increases (decreases) in precipitation in Mongolia and southern Europe are more obvious. The CAMO dipole has been strengthened since the LIA. The CAMO dipole is positively correlated with solar radiation and Northern Hemisphere temperature, and negatively correlated with Pacific decadal oscillations (PDO). Full article

Changes in the Earth’s climate and weather continue to impact the planet’s ecosystems, including the interface of infectious disease agents with their hosts and vectors. Environmental disasters, natural and human-made activities raise risk factors that indirectly facilitate infectious disease outbreaks. Subsequently, changes in habitat, displaced populations, and environmental stresses that affect the survival of species are amplified over time. The recurrence and spread of vector-borne (e.g., mosquito, tick, aphid) human, animal, and plant pathogens to new geographic locations are also influenced by climate change. The distribution and range of humans, agricultural animals and plants, wildlife and native plants, as well as vectors, parasites, and microbes that cause neglected diseases of the tropics as well as other global regions are also impacted. In addition, genomic sequencing can now be applied to detect signatures of infectious pathogens as they move into new regions. Molecular detection assays complement metagenomic sequencing to help us understand the microbial community found within the microbiomes of hosts and vectors, and help us uncover mechanistic relationships between climate variability and pathogen transmission. Our understanding of, and responses to, such complex dynamics and their impacts can be enhanced through effective, multi-sectoral One Health engagement coupled with applications of both traditional and novel technologies. Concerted efforts are needed to further harness and leverage technology that can identify and track these impacts of climate changes in order to mitigate and adapt to their effects. Full article

The purpose of the study presented in the paper is to highlight the influence of the microeconomic factors related to the evolutionary stage of the industry’s life cycle on the industry dynamics. The authors use the example of the Russian tourism industry to show that microeconomic factors are important, along with the macroeconomic, market, and demand characteristics external to the industry. Data mining was applied to obtain data from the industrial enterprise database and Rostourism official documents since there are no regular Russian statistics on firms’ exit and new entry. The authors used annual ranked listing of firms by their revenues to determine the structural indicators of the industry. The results confirm that it is important to consider not only the demand and macroeconomic indicators, which are external risks in relation to the industry, but also the internal processes at the different stages of the product cycle. In a sufficiently long period, the influence of microeconomic indicators may be no less strong than the business factors of financial risk. One should take this into consideration in econometric modeling on long time-series. Full article

Fish is a major part of the Bangladeshi diet, but data on the nutrient composition of marine fish species are sparse. Mesopelagic fish may be a new potential resource of food and nutrients; however, nutrient composition data are lacking. The aim of this study was to provide nutrient composition data of fish species sampled off the coast of Bangladesh and determine their potential contribution to recommended nutrient intakes (RNI). Seven species from the pelagic, mesopelagic, and demersal zones were sampled from the coast of Bangladesh with Dr. Fridtjof Nansen in 2018. Three pooled samples containing 15-840 individuals from each species were analysed at the Institute of Marine Research, Norway. The demersal species contained substantially lower concentrations of nearly all nutrients, whereas the mesopelagic species generally were more nutrient dense. All species, except for the demersal species Bombay duck (9% dry matter), were found to contribute ≥100% to the RNI of vitamin B₁₂, eicosapentaenoic acid, docosahexaenoic acid, and selenium. All species, except for the demersal fish species, contributed ≥25% to the RNI of six or more nutrients. The data presented in this paper are an important contribution to the Bangladeshi food composition table and contribute to the understanding of fish as an important source of micronutrients. Full article

Time to acceptance from submission and time to publication (publication lag) determines how quickly novel information is made available to other scientists and experts. In the medical field, the review process and revisions usually takes 3–4 months; the total time from submission to publication is 8–9 months. During the COVID-19 pandemic, information should be available much faster. The analysis of 833 documents published on SARS-CoV-2 and COVID-19 prior to 19 March 2020 shows that these times shrunk by a factor of ten. The median time to acceptance was three days for all publications, six days for research papers and reviews, four days for case studies and two days for other publication types. The median publication lag was nine days for all publications together, 11 days for research papers, nine days for case studies, 13 days for reviews and seven days for other publications. This demonstrates that the publication process—if necessary—can be sped up. For the sake of scientific accuracy, review times should not be pushed down, but the time from acceptance to actual publication could be shorter. Full article

This paper presents a comparison of the impact of milling technology in the computer numerically controlled (CNC) machining centre and selective laser sintering (SLS) and on the structure and properties of solid Ti6Al4V alloy. It has been shown that even small changes in technological conditions in the SLS manufacturing variant significantly affect changes from two to nearly two and a half times in tensile and bending strengths. Both the tensile and bending strength obtained in the most favourable manufacturing variant by the SLS method is over 25% higher than in the case of cast materials subsequently processed by milling. Plug-and-play SLS conditions provide about 60% of the possibilities. Structural, tribological and electrochemical tests were carried out. In vitro biological tests using osteoblasts confirm the good tendency for the proliferation of live cells on the substrate manufactured under the most favourable SLS conditions. The use of SLS additive technology for the manufacturing of dental implants and abutments made of Ti6Al4V alloy in combination with the digitisation of dental diagnostics and computer-aided design and manufacture of computer-aided design/manufacturing (CAD/CAM) following the idea of Dentistry 4.0 is the best choice of technology for manufacturing of prosthetic and implant devices used in dentistry. Full article

We investigate the state dependence of the variance of the instantaneous variance of the S&P 500 index empirically. Time-series analysis of realized variance over a 20-year period shows strong evidence of an elasticity of variance of the variance parameter close to that of a log-normal model, albeit with an empirical autocorrelation function that one-factor diffusion models fail to capture at horizons above a few weeks. When studying option market behavior (in-sample pricing as well as out-of-sample pricing and hedging over the period 2004–2019), messages are mixed, but systematic, model-wise. The log-normal but drift-free SABR (stochastic-alpha-beta-rho) model performs best for short-term options (times-to-expiry of three months and below), the Heston model—in which variance is stationary but not log-normal—is superior for long-term options, and a mixture of the two models does not lead to improvements. Full article

Models of set theory are defined, in which nonconstructible reals first appear on a given level of the projective hierarchy. Our main results are as follows. Suppose that $n\ge 2$ . Then: 1. If it holds in the constructible universe $\mathbf{L}$ that $a\subseteq \omega$ and $a\notin {\Sigma }_n^1\cup {\Pi }_n^1$ , then there is a generic extension of $\mathbf{L}$ in which $a\in {\Delta }_{n+1}^1$ but still $a\notin {\Sigma }_n^1\cup {\Pi }_n^1$ , and moreover, any set $x\subseteq \omega$ , $x\in {\Sigma }_n^1$ , is constructible and ${\Sigma }_n^1$ in $\mathbf{L}$ . 2. There exists a generic extension $\mathbf{L}$ in which it is true that there is a nonconstructible ${\Delta }_{n+1}^1$ set $a\subseteq \omega$ , but all ${\Sigma }_n^1$ sets $x\subseteq \omega$ are constructible and even ${\Sigma }_n^1$ in $\mathbf{L}$ , and in addition, $\mathbf{V}=\mathbf{L}\left[a\right]$ in the extension. 3. There exists an generic extension of $\mathbf{L}$ in which there is a nonconstructible ${\Sigma }_{n+1}^1$ set $a\subseteq \omega$ , but all ${\Delta }_{n+1}^1$ sets $x\subseteq \omega$ are constructible and ${\Delta }_{n+1}^1$ in $\mathbf{L}$ . Thus, nonconstructible reals (here subsets of $\omega$ ) can first appear at a given lightface projective class strictly higher than ${\Sigma }_2^1$ , in an appropriate generic extension of $\mathbf{L}$ . The lower limit ${\Sigma }_2^1$ is motivated by the Shoenfield absoluteness theorem, which implies that all ${\Sigma }_2^1$ sets $a\subseteq \omega$ are constructible. Our methods are based on almost-disjoint forcing. We add a sufficient number of generic reals to $\mathbf{L}$ , which are very similar at a given projective level n but discernible at the next level $n+1$ . Full article

The subshift of finite type property (also known as the Markov property) is ubiquitous in dynamical systems and the simplest and most widely studied class of dynamical systems are $\beta$ -shifts, namely transformations of the form $T_{\beta ,\alpha }:x\mapsto \beta x+\alpha mod1$ acting on $[-\alpha /(\beta -1),(1-\alpha )/(\beta -1)]$ , where $(\beta ,\alpha )\in \Delta$ is fixed and where $\Delta ≔\{(\beta ,\alpha )\in {\mathbb{R}}^2:\beta \in (1,2)\mathrm{and}0\le \alpha \le 2-\beta \}$ . Recently, it was shown, by Li et al. (Proc. Amer. Math. Soc. 147(5): 2045–2055, 2019), that the set of $(\beta ,\alpha )$ such that $T_{\beta ,\alpha }$ has the subshift of finite type property is dense in the parameter space $\Delta$ . Here, they proposed the following question. Given a fixed $\beta \in (1,2)$ which is the n-th root of a Perron number, does there exists a dense set of $\alpha$ in the fiber $\left\{\beta \right\}\times (0,2-\beta )$ , so that $T_{\beta ,\alpha }$ has the subshift of finite type property? We answer this question in the positive for a class of Pisot numbers. Further, we investigate if this question holds true when replacing the subshift of finite type property by the sofic property (that is a factor of a subshift of finite type). In doing so we generalise, a classical result of Schmidt (Bull. London Math. Soc., 12(4): 269–278, 1980) from the case when $\alpha =0$ to the case when $\alpha \in (0,2-\beta )$ . That is, we examine the structure of the set of eventually periodic points of $T_{\beta ,\alpha }$ when $\beta$ is a Pisot number and when $\beta$ is the n-th root of a Pisot number. Full article

The necessity of coordination among entities is essential for the success of any supply chain management (SCM). This paper focuses on coordination between two players and cost-sharing in an SCM that considers a vendor and a buyer. For random demand and complex product production, a flexible production system is recommended. The study aims to minimize the total SCM cost under stochastic conditions. In the flexible production systems, the production rate is introduced as the decision variable and the unit production cost is minimum at the obtained optimal value. The setup cost of flexible systems is higher and to control this, a discrete investment function is utilized. The exact information about the probability distribution of lead time demand is not available with known mean and variance. The issue of unknown distribution of lead time demand is solved by considering a distribution-free approach to find the amount of shortages. The game-theoretic approach is employed to obtain closed-form solutions. First, the model is solved under decentralized SCM based on the Stackelberg model, and then solved under centralized SCM. Bargaining is the central theme of any business nowadays among the players of an SCM to make their profit within a centralized and decentralized setup. For this, a cost allocation model for lead time crashing cost based on the Nash bargaining model with the satisfaction level of SCM members is proposed. The cost allocation model under Nash bargaining achieves exciting results in SCM coordination. Full article

Heterostyly employs distinct hermaphroditic floral morphs to enforce outbreeding. Morphs differ structurally in stigma/anther positioning, promoting cross-pollination, and physiologically blocking self-fertilization. Heterostyly is controlled by a self-incompatibility (S)-locus of a small number of linked S-genes specific to short-styled morph genomes. Turnera possesses three S-genes, namely TsBAHD (controlling pistil characters), TsYUC6, and TsSPH1 (controlling stamen characters). Here, we compare pistil and stamen transcriptomes of floral morphs of T. subulata to investigate hypothesized S-gene function(s) and whether hormonal differences might contribute to physiological incompatibility. We then use network analyses to identify genetic networks underpinning heterostyly. We found a depletion of brassinosteroid-regulated genes in short styled (S)-morph pistils, consistent with hypothesized brassinosteroid-inactivating activity of TsBAHD. In S-morph anthers, auxin-regulated genes were enriched, consistent with hypothesized auxin biosynthesis activity of TsYUC6. Evidence was found for auxin elevation and brassinosteroid reduction in both pistils and stamens of S- relative to long styled (L)-morph flowers, consistent with reciprocal hormonal differences contributing to physiological incompatibility. Additional hormone pathways were also affected, however, suggesting S-gene activities intersect with a signaling hub. Interestingly, distinct S-genes controlling pistil length, from three species with independently evolved heterostyly, potentially intersect with phytochrome interacting factor (PIF) network hubs which mediate red/far-red light signaling. We propose that modification of the activities of PIF hubs by the S-locus could be a common theme in the evolution of heterostyly. Full article

A Bayesian framework is presented for finite element model-updating using experimental modal data. A novel likelihood formulation is proposed regarding the inclusion of the mode shapes, based on a probabilistic treatment of the MAC value between the model predicted and experimental mode shapes. The framework is demonstrated by performing model-updating for the Metsovo bridge using a reduced high-fidelity finite element model. Experimental modal identification methods are used in order to extract the modal characteristics of the bridge from ambient acceleration time histories obtained from field measurements exploiting a network of reference and roving sensors. The Transitional Markov Chain Monte Carlo algorithm is used to perform the model updating by drawing samples from the posterior distribution of the model parameters. The proposed framework yields reasonable uncertainty bounds for the model parameters, insensitive to the redundant information contained in the measured data due to closely spaced sensors. In contrast, conventional Bayesian formulations which use probabilistic models to characterize the components of the discrepancy vector between the measured and model-predicted mode shapes result in unrealistically thin uncertainty bounds for the model parameters for a large number of sensors. Full article

In two-stage single-phase inverters, inherent double line frequency ripple is present at both the input and output of the front-end converter. Generally, large electrolytic capacitors are used to eliminate this double line frequency ripple. It is well known that low frequency ripple shortens the lifespan of capacitors. Hence, the system reliability can get worse. In order to eliminate the double line frequency ripple, additional hardware combined with an energy storage device is required in most of the methods developed so far. In this paper, a novel power-decoupling control method is proposed to eliminate the double line frequency ripple at the front-end converter of two-stage single phase DC/AC power conversion systems. The proposed control algorithm is composed of two loops, a ripple compensation loop and an average voltage control loop, and no extra hardware is required. Since the proposed method does not require information from the phase-locked-loop (PLL) of the inverter, it is independent of inverter control. In order to verify the validity and feasibility of the proposed algorithm a 5 kW Dual Active Bridge (DAB) DC/DC converter and a single-phase inverter are implemented. The effectiveness of the proposed method is verified through the simulation and experimental results. Full article

Background: The identification of C-repeat binding factor (CBF), and the characterization as an inducer of CBF Expression 1 (ICE1), and a major activator for C-repeat binding factor, were important breakthroughs in the cold signaling network. Methods: In the present study, the full length cDNA of ICE1 was isolated from the tea tree (Camellia sinensis). CsICE1 protein was located in the cell nucleus as revealed by subcellular localization analysis. To investigate the biological functions of CsICE1, a transgenic line fused with the CsICE1 gene in Arabidopsis thaliana (arabidopsis) was generated by the floral dip method. Results: The CsICE1 was expressed differentially in various tea tree tissues, mostly in buds and leaves, and the transcript level of CsICE1 was increased after 1 h and peaked at 2 h under cold treatment. Transcription activity assay indicated that the spermine synthase (SPMS) and arginine decarboxylase (ADC) genes were possible targets of CsICE1. In addition, the values of net photosynthetic rate, transpiration rate, stomatal conductance in transgenic lines declined by less extent than wild-type plants under low temperatures. Furthermore, transcript levels of ADC genes in the transgenic lines had no apparent alteration under normal growth conditions but substantially increased under cold conditions, consistent of changes in free polyamine levels. Taken together, these results demonstrated that CsICE1 plays a positive role in cold tolerance, which may be due to the modulation of polyamine levels through interacting with CsADC. Full article