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Abstract Background Cell differentiation requires the integration of two opposite processes, a stabilizing cellular memory, especially at the transcriptional scale, and a burst of gene expression variability which follows the differentiation induction. Therefore, the actual capacity of a cell to undergo phenotypic change during a differentiation process relies upon a modification in this balance which favors change-inducing gene expression variability. However, there are no experimental data providing insight on how fast the transcriptomes of identical cells would diverge on the scale of the very first two cell divisions during the differentiation process. Results In order to quantitatively address this question, we developed different experimental methods to recover the transcriptomes of related cells, after one and two divisions, while preserving the information about their lineage at the scale of a single cell division. We analyzed the transcriptomes of related cells from two differentiation biological systems (human CD34+ cells and T2EC chicken primary erythrocytic progenitors) using two different single-cell transcriptomics technologies (scRT-qPCR and scRNA-seq). Conclusions We identified that the gene transcription profiles of differentiating sister cells are more similar to each other than to those of non-related cells of the same type, sharing the same environment and undergoing similar biological processes. More importantly, we observed greater discrepancies between differentiating sister cells than between self-renewing sister cells. Furthermore, a progressive increase in this divergence from first generation to second generation was observed when comparing differentiating cousin cells to self renewing cousin cells. Our results are in favor of a gradual erasure of transcriptional memory during the differentiation process.

The current study examines associations between mindful parenting, family support, and youth information management practices (disclosure, lying, and secrecy) in a diverse sample of college-attending emerging adults (n = 304). Participants were recruited through college classes, and students completed surveys in classrooms or in nearby campus spaces. Data were analyzed to assess for the potential interactive implications of parental mindfulness and support. Results indicate that both parenting variables are important to consider when conceptualizing emerging adults’ information management. Youth who reported the highest levels of parental mindfulness and support also disclosed the most to their parents about their daily activities. In contrast, youth who reported the lowest levels of mindfulness and support lied the most to their parents. Low support was also associated with increased secrecy. Results suggest that parent-youth relationships continue to be important through the emerging adult developmental period, especially in terms of promoting communication practices that can help youth adjust to their increasingly complex worlds.

Abstract Many of the world's most important crops are polyploid. The presence of more than two sets of chromosomes within their nuclei and frequently aberrant reproductive biology in polyploids present obstacles to conventional breeding. The presence of a larger number of homoeologous copies of each gene makes random mutation breeding a daunting task for polyploids. Genome editing has revolutionized improvement of polyploid crops as multiple gene copies and/or alleles can be edited simultaneously while preserving the key attributes of elite cultivars. Most genome‐editing platforms employ sequence‐specific nucleases (SSNs) to generate DNA double‐stranded breaks at their target gene. Such DNA breaks are typically repaired via the error‐prone nonhomologous end‐joining process, which often leads to frame shift mutations, causing loss of gene function. Genome editing has enhanced the disease resistance, yield components, and end‐use quality of polyploid crops. However, identification of candidate targets, genotyping, and requirement of high mutagenesis efficiency remain bottlenecks for targeted mutagenesis in polyploids. In this review, we will survey the tremendous progress of SSN‐mediated targeted mutagenesis in polyploid crop improvement, discuss its challenges, and identify optimizations needed to sustain further progress.

DIY's coastal area is one of DIY's development priorities by establishing the site as an economic corridor that impacts physical development, such as settlements in the coastal area of DIY. This study aims to analyze the distribution and density of settlements. The method used in research is a quantitative descriptive and Geography Information System approach. The analysis used in this study is Nearest Neighbour Analysis (NNA) and Kernel Density Analysis with an analysis tool in ArcGIS 10.5. The results of the NNA show that the three districts located on the coast of DIY have the same distribution pattern characteristics, namely, the clustered pattern, which means the NNA index value is

ZnO-Au nanocomposite thin films have been previously reported as hybrid metamaterials with unique optical properties such as plasmonic resonance properties and hyperbolic behaviors. In this study, Au composition in the ZnO-Au nanocomposites has been effectively tuned by target composition variation and thus resulted in microstructure and optical property tuning. Specifically, all the ZnO-Au nanocomposite thin films grown through the pulsed laser deposition (PLD) method show obvious vertically aligned nanocomposite (VAN) structure with the Au nanopillars embedded in the ZnO matrix. Moreover, the average diameter of Au nanopillars increases as Au concentration increases, which also leads to the redshifts in the surface plasmon resonance (SPR) wavelength and changes in the hyperbolic behaviors of the films. As a whole, this work discusses how strain-driven tuning of optical properties and microstructure resulted through a novel Au concentration variation approach which has not been previously attempted in the ZnO-Au thin film system. These highly ordered films present great promise in the areas of sensing, waveguides, and nanophotonics to name a few.

Summary: Background: Dengue is a global problem that seems to be worsening, as hyper-urbanization associated with climate change has led to a significant increase in the abundance and geographical spread of its principal vector, the Aedes aegypti mosquito. Currently available solutions have not been able to stop the spread of dengue which shows the urgent need to implement alternative technologies as practical solutions. In a previous pilot trial, we demonstrated the efficacy and safety of the method ‘Natural Vector Control’ (NVC) in suppressing the Ae. aegypti vector population and in blocking the occurrence of an outbreak of dengue in the treated areas. Here, we expand the use of the NVC program in a large-scale 20 months intervention period in an entire city in southern Brazil. Methods: Sterile male mosquitoes were produced from locally sourced Ae. aegypti mosquitoes by using a treatment that includes double-stranded RNA and thiotepa. Weekly massive releases of sterile male mosquitoes were performed in predefined areas of Ortigueira city from November 2020 to July 2022. Mosquito monitoring was performed by using ovitraps during the entire intervention period. Dengue incidence data was obtained from the Brazilian National Disease Surveillance System. Findings: During the two epidemiological seasons, the intervention in Ortigueira resulted in up to 98.7% suppression of live progeny of field Ae. aegypti mosquitoes recorded over time. More importantly, when comparing the 2020 and 2022 dengue outbreaks that occurred in the region, the post-intervention dengue incidence in Ortigueira was 97% lower compared to the control cities. Interpretation: The NVC method was confirmed to be a safe and efficient way to suppress Ae. aegypti field populations and prevent the occurrence of a dengue outbreak. Importantly, it has been shown to be applicable in large-scale, real-world conditions. Funding: This study was funded by Klabin S/A and Forrest Innovations Ltd.

Cell lineage tracking is a long-standing and unresolved problem in biology. Microfluidic technologies have the potential to address this problem, by virtue of their ability to manipulate and process single-cells in a rapid, controllable and efficient manner. Indeed, when coupled with traditional imaging approaches, microfluidic systems allow the experimentalist to follow single-cell divisions over time. Herein, we present a valve-based microfluidic system able to probe the decision-making processes of single-cells, by tracking their lineage over multiple generations. The system operates by trapping single-cells within growth chambers, allowing the trapped cells to grow and divide, isolating sister cells after a user-defined number of divisions and finally extracting them for downstream transcriptome analysis. The platform incorporates multiple cell manipulation operations, image processing-based automation for cell loading and growth monitoring, reagent addition and device washing. To demonstrate the efficacy of the microfluidic workflow, 6C2 (chicken erythroleukemia) and T2EC (primary chicken erythrocytic progenitors) cells are tracked inside the microfluidic device over two generations, with a cell viability rate in excess of 90%. Sister cells are successfully isolated after division and extracted within a 500 nL volume, which was demonstrated to be compatible with downstream single-cell RNA sequencing analysis.

When human cord blood-derived CD34+ cells are induced to differentiate, they undergo rapid and dynamic morphological and molecular transformations that are critical for fate commitment. In particular, the cells pass through a transitory phase known as "multilineage-primed" state. These cells are characterized by a mixed gene expression profile, different in each cell, with the coexpression of many genes characteristic for concurrent cell lineages. The aim of our study is to understand the mechanisms of the establishment and the exit from this transitory state. We investigated this issue using single-cell RNA sequencing and ATAC-seq. Two phases were detected. The first phase is a rapid and global chromatin decompaction that makes most of the gene promoters in the genome accessible for transcription. It results 24 h later in enhanced and pervasive transcription of the genome leading to the concomitant increase in the cell-to-cell variability of transcriptional profiles. The second phase is the exit from the multilineage-primed phase marked by a slow chromatin closure and a subsequent overall down-regulation of gene transcription. This process is selective and results in the emergence of coherent expression profiles corresponding to distinct cell subpopulations. The typical time scale of these events spans 48 to 72 h. These observations suggest that the nonspecificity of genome decompaction is the condition for the generation of a highly variable multilineage expression profile. The nonspecific phase is followed by specific regulatory actions that stabilize and maintain the activity of key genes, while the rest of the genome becomes repressed again by the chromatin recompaction. Thus, the initiation of differentiation is reminiscent of a constrained optimization process that associates the spontaneous generation of gene expression diversity to subsequent regulatory actions that maintain the activity of some genes, while the rest of the genome sinks back to the repressive closed chromatin state.

Background Since 2010, Israel has expanded the adoption of procedure-related group (PRG) based payments for hospitals. While there is a rich quantitative literature that assesses the effects of payment reforms on efficiency or quality of care, very few qualitative studies have focused on the impacts of diagnosis-related group (DRG)-like payments on hospitals from the perspective of hospital workers as change agents. Methods We used a qualitative, thematic analysis based on 33 semi-structured in-depth interviews with chief executive officers (CEOs), chief financial officers (CFOs), ward directors and physicians conducted in five public hospitals in Israel, sampled by maximum variation according to hospital characteristics. Results Interviewees reported that the payment reform led to organizational changes such as increased transparency and enhanced supervision. Interviewees also reported several actions in response to the economic incentives of PRGbased payment. These included (1) shifting activities to afterhours and using operating rooms (ORs) more efficiently to enable increased surgical volumes; (2) reducing costs by shortening lengths of stay and increasing cost-consciousness in procurement; and (3) increasing revenues by improving coding and selecting procedures. Moderating factors reduced the effects of the reform. For example, organizational factors such as the public nature of hospitals or the (un)availability of healthcare resources did not always allow hospitals to increase the number of cases treated. Also, conflicting incentives such as multiple payment mechanisms or underpricing of procedures blurred the incentives of the reform. Finally, managers and physicians have many other considerations that outweigh the economic ones. Conclusion PRG payments affected the organizational dynamics of hospitals and changed decision-making about admission and treatment policies. However, such effects were moderated by many other factors that should be considered when shaping and analyzing hospital payment reforms.

Penelitian ini bertujuan untuk mendeskripsikan keterlaksanaan standar proses pada pembelajaran matematika menurut Kurikulum 2013 pada kelas XII SMA Negeri. Penelitian ini merupakan penelitian mixed method dengan desain sequential explanatory strategy. Partisipan pada penelitian ini adalah guru matematika kelas XII SMA Negeri di Kota Pekanbaru yang menerapkan Kurikulum 2013. Keterlaksanaan standar proses dinilai berdasarkan aspek perencanaan, pelaksanaan, dan penilaian pembelajaran. Analisis data deskriptif menggunakan kecenderungan keterlaksanaan dalam lima kriteria, yaitu sangat baik, baik, cukup, kurang, dan sangat kurang. Hasil penelitian menunjukkan bahwa keterlaksanaan standar proses menurut Kurikulum 2013 pada kelas XII SMA Negeri di Kota Pekanbaru ditinjau dari aspek perencanaan, pelaksanaan, dan penilaian pembelajaran adalah masuk pada kategori baik. Hasil penelitian juga menunjukkan bahwa pada aspek perencanaan guru masih mengalami kendala dalam penyusunan lembar kegiatan siswa, rencana pelaksanaan pembelajaran, serta perencanaan alokasi waktu. Pada aspek pelaksanaan pembelajaran, masih terdapat kendala yaitu pada kemauan guru untuk menerapkan pendekatan saintifik, terutama pada tahap menanya dan menalar. Adapun pada aspek penilaian, guru masih mengalami kendala terkait penilaian sikap dan keterampilan. The implementation of process standard in mathematics learning according to the Curriculum 2013 in 12th-grade of public senior high school Abstract This study aimed to describe the implementation of process standard in mathematics learning according to the Curriculum 2013 in 12th-grade of public senior high school. This research was mixed-method research with a sequential explanatory strategy design. Participants in this study were mathematics teachers of 12th-grade of public senior high school in Pekanbaru City, Indonesia who applied the Curriculum 2013. The process standard was assessed based on aspects of planning, implementing, and assessment of learning. Descriptive data analysis used the tendency of implementation in five criteria, namely very good, good, enough, less, and very less. The results showed that the implementation of process standard according to the Curriculum 2013 in class 12th-grade of public senior high school in Pekanbaru City in terms of planning, implementation, and assessment of learning were included in the good category. The results of the study also showed that in the aspect of planning, the teacher was still experiencing problems in the preparation of students’ worksheet, lesson plan, and time allocation planning. In the aspect of implementing learning, there were still obstacles, namely the teacher's willingness to apply a scientific approach, especially at the questioning and reasoning stages. As for the aspects of assessment, teachers still experience obstacles related to the assessment of attitudes and skills.

Nanocomposite thin film materials present great opportunities in coupling materials and functionalities in unique nanostructures including nanoparticles-in-matrix, vertically aligned nanocomposites (VANs), and nanolayers. Interestingly the nanocomposites processed through a non-equilibrium processing method, e.g., pulsed laser deposition (PLD), often possess unique metastable phases and microstructures that could not achieve using equilibrium techniques, and thus lead to novel physical properties. In this work, a unique three-phase system composed of BaTiO3 (BTO), with two immiscible metals, Au and Fe, is demonstrated. By adjusting the deposition laser frequency from 2 Hz to 10 Hz, the phase and morphology of Au and Fe nanoparticles in BTO matrix vary from separated Au and Fe nanoparticles to well-mixed Au-Fe alloy pillars. This is attributed to the non-equilibrium process of PLD and the limited diffusion under high laser frequency (e.g., 10 Hz). The magnetic and optical properties are effectively tuned based on the morphology variation. This work demonstrates the stabilization of non-equilibrium alloy structures in the VAN form and allows for the exploration of new non-equilibrium materials systems and their properties that could not be easily achieved through traditional equilibrium methods.

Epitaxial lithium niobate (LNO) thin films are an attractive material for devices across a broad range of fields, including optics, acoustics, and electronics. These applications demand high‐quality thin films without in‐plane growth domains to reduce the optical/acoustical losses and optimize efficiency. Twin‐free single‐domain‐like growth has been achieved previously, but it requires specific growth conditions that might be hard to replicate. In this work, a versatile nanocomposite‐seeded approach is demonstrated as an effective approach to grow single‐domain epitaxial lithium niobate thin films. Films are grown through a pulsed laser deposition method and growth conditions are optimized to achieve high‐quality epitaxial film. A comprehensive microstructure characterization is performed and optical properties are measured. A piezoelectric acoustic resonator device is developed to demonstrate the future potential of the nanocomposite‐seeded approach for high‐quality LNO growth for radio frequency (RF) applications.

Hyperbolic metamaterials are a class of materials exhibiting anisotropic dielectric function owing to the morphology of the nanostructures. In these structures, one direction behaves as a metal, and the orthogonal direction behaves as a dielectric material. Applications include subdiffraction imaging and hyperlenses. However, key limiting factors include energy losses of noble metals and challenging fabrication methods. In this work, self-assembled plasmonic metamaterials consisting of anisotropic nanoalloy pillars embedded into the ZnO matrix are developed using a seed-layer approach. Alloys of AuxAl1−x or AuxCu1−x are explored due to their lower losses and higher stability. Optical and microstructural properties were explored. The ZnO-AuxCu1−x system demonstrated excellent epitaxial quality and optical properties compared with the ZnO-AuxAl1−x system. Both nanocomposite systems demonstrate plasmonic resonance, hyperbolic dispersion, low losses, and epsilon-near-zero permittivity, making them promising candidates towards direct photonic integration.

Oxytocin (OT) and vasopressin (AVP) are neuropeptides that govern the social-emotional functioning of humans. We contend that to fully understand their function, research should consider how they are flexibly fitted to maximize survival and reproduction given the variety of human experience. In a series of two studies, we show that early life stress is associated with change in the core function of OT and AVP in evolutionary predictable ways: Under high early life stress, AVP promotes threat-detection capabilities, whereas OT motivates non-selective proximity seeking to others. Conversely, under low early life stress these neuropeptides have an opposite, yet adaptive response: AVP promotes low vigilance and preservation of energy, whereas OT increases detection of interpersonal flaws. Our results demonstrate the plasticity of neuropeptide functioning that mirrors the variance in human social-emotional functioning.

Disasters caused losses and damage to the coastal areas of Pariaman City. The construction costs for development in coastal areas are relatively expensive. Early good research must be done to monitor shoreline changes and reduce losses and damage in the coastal areas of Pariaman City. The method used to monitor shoreline changes using satellite images with GIS techniques. The coastal change analyzes is studied including coastal erosion and sedimentation. In this study, disaster mitigation and impacts reduction caused by coastal changes are the final purposes. The losses and damage due to coastal changes were monitored during a certain time. The results of this study are aimed to make a solution for disaster mitigation in coastal areas caused by the shoreline changes in Pariaman City.

Despite the stochastic noise that characterizes all cellular processes the cells are able to maintain and transmit to their daughter cells the stable level of gene expression. In order to better understand this phenomenon, we investigated the temporal dynamics of gene expression variation using a double reporter gene model. We compared cell clones with transgenes coding for highly stable mRNA and fluorescent proteins with clones expressing destabilized mRNA-s and proteins. Both types of clones displayed strong heterogeneity of reporter gene expression levels. However, cells expressing stable gene products produced daughter cells with similar level of reporter proteins, while in cell clones with short mRNA and protein half-lives the epigenetic memory of the gene expression level was completely suppressed. Computer simulations also confirmed the role of mRNA and protein stability in the conservation of constant gene expression levels over several cell generations. These data indicate that the conservation of a stable phenotype in a cellular lineage may largely depend on the slow turnover of mRNA-s and proteins.

Epigenetic modifications may occur during in vitro manipulations of stem cells but these effects have remained unexplored in the context of cell and gene therapy protocols. In an experimental model of ex vivo gene modification for hematopoietic gene therapy, human CD34(+) cells were cultured shortly in the presence of cytokines then with a gene transfer lentiviral vector (LV) expected to transduce cells but to have otherwise limited biological effects on the cells. At the end of the culture, the population of cells remained largely similar at the phenotypic level but some epigenetic changes were evident. Exposure of CD34(+) cells to cytokines increased nuclear expression of epigenetic regulators SIRT1 or DNMT1 and caused genome-wide DNA methylation changes. Surprisingly, the LV caused additional and distinct effects. Large-scale genomic DNA methylation analysis showed that balanced methylation changes occurred in about 200 genes following culture of CD34(+) cells in the presence of cytokines but 900 genes were modified following addition of the LV, predominantly increasing CpG methylation. Epigenetic effects resulting from ex vivo culture and from the use of LV may constitute previously unsuspected sources of biological effects in stem cells and may provide new biomarkers to rationally optimize gene and cell therapy protocols.

The importance of honey bees to the world economy far surpasses their contribution in terms of honey production; they are responsible for up to 30% of the world's food production through pollination of crops. Since fall 2006, honey bees in the U.S. have faced a serious population decline, due in part to a phenomenon called Colony Collapse Disorder (CCD), which is a disease syndrome that is likely caused by several factors. Data from an initial study in which investigators compared pathogens in honey bees affected by CCD suggested a putative role for Israeli Acute Paralysis Virus, IAPV. This is a single stranded RNA virus with no DNA stage placed taxonomically within the family Dicistroviridae. Although subsequent studies have failed to find IAPV in all CCD diagnosed colonies, IAPV has been shown to cause honey bee mortality. RNA interference technology (RNAi) has been used successfully to silence endogenous insect (including honey bee) genes both by injection and feeding. Moreover, RNAi was shown to prevent bees from succumbing to infection from IAPV under laboratory conditions. In the current study IAPV specific homologous dsRNA was used in the field, under natural beekeeping conditions in order to prevent mortality and improve the overall health of bees infected with IAPV. This controlled study included a total of 160 honey bee hives in two discrete climates, seasons and geographical locations (Florida and Pennsylvania). To our knowledge, this is the first successful large-scale real world use of RNAi for disease control.

BACKGROUND: The spontaneous emergence of phenotypic heterogeneity in clonal populations of mammalian cells in vitro is a rule rather than an exception. We consider two simple, mutually non-exclusive models that explain the generation of diverse cell types in a homogeneous population. In the first model, the phenotypic switch is the consequence of extrinsic factors. Initially identical cells may become different because they encounter different local environments that induce adaptive responses. According to the second model, the phenotypic switch is intrinsic to the cells that may occur even in homogeneous environments. PRINCIPAL FINDINGS: We have investigated the "extrinsic" and the "intrinsic" mechanisms using computer simulations and experimentation. First, we simulated in silico the emergence of two cell types in a clonal cell population using a multiagent model. Both mechanisms produced stable phenotypic heterogeneity, but the distribution of the cell types was different. The "intrinsic" model predicted an even distribution of the rare phenotype cells, while in the "extrinsic" model these cells formed small clusters. The key predictions of the two models were confronted with the results obtained experimentally using a myogenic cell line. CONCLUSIONS: The observations emphasize the importance of the "ecological" context and suggest that, consistently with the "extrinsic" model, local stochastic interactions between phenotypically identical cells play a key role in the initiation of phenotypic switch. Nevertheless, the "intrinsic" model also shows some other aspects of reality: The phenotypic switch is not triggered exclusively by the local environmental variations, but also depends to some extent on the phenotypic intrinsic robustness of the cells.

There is increasing evidence in both plants and animals that epigenetic marks are not always cleared between generations. Incomplete erasure at genes associated with a measurable phenotype results in unusual patterns of inheritance from one generation to the next, termed transgenerational epigenetic inheritance. The Agouti viable yellow (A(vy)) allele is the best-studied example of this phenomenon in mice. The A(vy) allele is the result of a retrotransposon insertion upstream of the Agouti gene. Expression at this locus is controlled by the long terminal repeat (LTR) of the retrotransposon, and expression results in a yellow coat and correlates with hypomethylation of the LTR. Isogenic mice display variable expressivity, resulting in mice with a range of coat colours, from yellow through to agouti. Agouti mice have a methylated LTR. The locus displays epigenetic inheritance following maternal but not paternal transmission; yellow mothers produce more yellow offspring than agouti mothers. We have analysed the DNA methylation in mature gametes, zygotes, and blastocysts and found that the paternally and maternally inherited alleles are treated differently. The paternally inherited allele is demethylated rapidly, and the maternal allele is demethylated more slowly, in a manner similar to that of nonimprinted single-copy genes. Interestingly, following maternal transmission of the allele, there is no DNA methylation in the blastocyst, suggesting that DNA methylation is not the inherited mark. We have independent support for this conclusion from studies that do not involve direct analysis of DNA methylation. Haplo-insufficiency for Mel18, a polycomb group protein, introduces epigenetic inheritance at a paternally derived A(vy) allele, and the pedigrees reveal that this occurs after zygotic genome activation and, therefore, despite the rapid demethylation of the locus.