Online Inverse Optimal Control for Time-Varying Cost Weights.

Inverse optimal control is a method for recovering the cost function used in an optimal control problem in expert demonstrations. Most studies on inverse optimal control have focused on building the unknown cost function through the linear combination of given features with unknown cost weights, which are generally considered to be constant. However, in many real-world applications, the cost weights may vary over time. In this study, we propose an adaptive online inverse optimal control approach based on a neural-network approximation to address the challenge of recovering time-varying cost weights. We conduct a well-posedness analysis of the problem and suggest a condition for the adaptive goal, under which the weights of the neural network generated to achieve this adaptive goal are unique to the corresponding inverse optimal control problem. Furthermore, we propose an updating law for the weights of the neural network to ensure the stability of the convergence of the solutions. Finally, simulation results for an example linear system are presented to demonstrate the effectiveness of the proposed strategy. The proposed method is applicable to a wide range of problems requiring real-time inverse optimal control calculations.

VGLL1 cooperates with TEAD4 to control human trophectoderm lineage specification.

In contrast to rodents, the mechanisms underlying human trophectoderm and early placenta specification are understudied due to ethical barriers and the scarcity of embryos. Recent reports have shown that human pluripotent stem cells (PSCs) can differentiate into trophectoderm (TE)-like cells (TELCs) and trophoblast stem cells (TSCs), offering a valuable in vitro model to study early placenta specification. Here, we demonstrate that the VGLL1 (vestigial-like family member 1), which is highly expressed during human and non-human primate TE specification in vivo but is negligibly expressed in mouse, is a critical regulator of cell fate determination and self-renewal in human TELCs and TSCs derived from naïve PSCs. Mechanistically, VGLL1 partners with the transcription factor TEAD4 (TEA domain transcription factor 4) to regulate chromatin accessibility at target gene loci through histone acetylation and acts in cooperation with GATA3 and TFAP2C. Our work is relevant to understand primate early embryogenesis and how it differs from other mammalian species.

Different policies constrained agricultural non-point pollutants emission trading management for water system under interval, fuzzy, and stochastic information.

Formulating suitable policies is essential for resources and environmental management. In this study, an agricultural pollutants emission trading management model driven by water resources and pollutants control is developed to search reasonable policies for agricultural water resources allocation under multiple uncertainties. Random-fuzzy and interval information in water resources system that have directly impact on the effectiveness of management schemes is reflected through interval two-stage stochastic fuzzy-probability programming. The model was root from regional agricultural water resources system in Jining City, China under considering the relationship among effective precipitation, crop water demand, and pollutants emission. Two types policies (water consumption-control and pollutants emission-control) are designed for searching the related interaction on water resources management and water quality improvement. The results indicated that water resources policies would be of water and environmental double benefits, and a large rainfall would reduce irrigation amount from water sources and lead to a larger pollutants emission trading. The results will help for defining scientific and effective water resources protection and management policies and analyzing the related interacted effects on water consumption, pollutants control and system benefit.

Varying the expression pattern of the strigolactone receptor gene DAD2 results in phenotypes distinct from both wild type and knockout mutants.

The action of the petunia strigolactone (SL) hormone receptor DAD2 is dependent not only on its interaction with the PhMAX2A and PhD53A proteins, but also on its expression patterns within the plant. Previously, in a yeast-2-hybrid system, we showed that a series of a single and double amino acid mutants of DAD2 had altered interactions with these binding partners. In this study, we tested the mutants in two plant systems, Arabidopsis and petunia. Testing in Arabidopsis was enabled by creating a CRISPR-Cas9 knockout mutant of the Arabidopsis strigolactone receptor (AtD14). We produced SL receptor activity in both systems using wild type and mutant genes; however, the mutants had functions largely indistinguishable from those of the wild type. The expression of the wild type DAD2 from the CaMV 35S promoter in dad2 petunia produced plants neither quite like the dad2 mutant nor the V26 wild type. These plants had greater height and leaf size although branch number and the plant shape remained more like those of the mutant. These traits may be valuable in the context of a restricted area growing system such as controlled environment agriculture.

Two-Dimensional Peripheral Refraction and Higher-Order Wavefront Aberrations Induced by Orthokeratology Lenses Decentration.

Purpose: The purpose of this study was to explore two-dimensional peripheral refraction and higher-order aberrations (HOAs) induced by orthokeratology lens decentration. Methods: Two-dimensional peripheral refraction and HOAs in a rectangular field (horizontally 60 degrees and vertically 36 degrees) were obtained using an open-view Hartmann-Shack wavefront sensor. The peripheral field was divided into 8 regions according to a combination of superior (UZ) or inferior (LZ) and a value, 1 (T25 to T30), 2 (T20 to T25), 3 (N20 to N25), or 4 (N25 to N30). The decentration of the lens was evaluated based on the change of power in the front of the tangential corneal map. All measurements were taken at the baseline and 1 month after lens fitting. Results: In total, 134 myopic children (age = 12.47 ± 1.70 years, SER = -2.44 ± 1.10 diopters [D]) were recruited. In general, horizontally asymmetrical change was observed in relative peripheral refraction (RPR), spherical aberration (SA), and horizontal coma. The root-mean square of higher order aberration (RMSHOA) and vertical coma demonstrated radial symmetrical change and vertically asymmetric change, respectively. Relative peripheral myopia was significantly increased after the treatment, with more myopic refraction in the temporal side. RPR changes in UZ2, UZ3, UZ4, LZ1, and LZ2 were related to the amount of lens decentration (r ≈ 0.4, P < 0.05). All HOAs increased after lens fitting (around 0.03 um, 0.02 um, 0.04 um, and 0.41 um for SA, horizontal COMA, vertical COMA, and RMSHOA in the periphery region). Conclusions: RPR and HOAs are related to lens decentration, which might contribute to the efficacy of orthokeratology. Translational Relevance: The study found a decentration-related optical feature after 1 month of lens wear, which is a suggested protective factor in myopia treatment. The findings might provide new insights for customized contact lens myopia treatment based on optics.

Transcriptomic analysis implicates ABA signaling and carbon supply in the differential outgrowth of petunia axillary buds.

BACKGROUND: Shoot branching of flowering plants exhibits phenotypic plasticity and variability. This plasticity is determined by the activity of axillary meristems, which in turn is influenced by endogenous and exogenous cues such as nutrients and light. In many species, not all buds on the main shoot develop into branches despite favorable growing conditions. In petunia, basal axillary buds (buds 1-3) typically do not grow out to form branches, while more apical axillary buds (buds 6 and 7) are competent to grow. RESULTS: The genetic regulation of buds was explored using transcriptome analyses of petunia axillary buds at different positions on the main stem. To suppress or promote bud outgrowth, we grew the plants in media with differing phosphate (P) levels. Using RNA-seq, we found many (> 5000) differentially expressed genes between bud 6 or 7, and bud 2. In addition, more genes were differentially expressed when we transferred the plants from low P to high P medium, compared with shifting from high P to low P medium. Buds 6 and 7 had increased transcript abundance of cytokinin and auxin-related genes, whereas the basal non-growing buds (bud 2 and to a lesser extent bud 3) had higher expression of strigolactone, abscisic acid, and dormancy-related genes, suggesting the outgrowth of these basal buds was actively suppressed. Consistent with this, the expression of ABA associated genes decreased significantly in apical buds after stimulating growth by switching the medium from low P to high P. Furthermore, comparisons between our data and transcriptome data from other species suggest that the suppression of outgrowth of bud 2 was correlated with a limited supply of carbon to these axillary buds. Candidate genes that might repress bud outgrowth were identified by co-expression analysis. CONCLUSIONS: Plants need to balance growth of axillary buds into branches to fit with available resources while allowing some buds to remain dormant to grow after the loss of plant parts or in response to a change in environmental conditions. Here we demonstrate that different buds on the same plant with different developmental potentials have quite different transcriptome profiles.

Characteristics of the Ocular Surface in Myopic Child Candidates of Orthokeratology Lens Wear.

INTRODUCTION: The purpose of this study was to investigate the characteristics of objective ocular surface parameters using non-invasive objective instruments in children with myopia who are candidates for orthokeratology lens wear. METHODS: Children with myopia who are candidates for orthokeratology lens wear were retrospectively investigated. The subjects were divided into three age groups. The Keratograph 5M and LipiView interferometry were used to assess non-invasive tear meniscus height (TMH), non-invasive tear film break-up time (NIBUT), conjunctival hyperemia redness score (RS), meibomian gland loss (MGL) score, lipid layer thickness (LLT), and blink pattern analysis, including the number of partial blinks (PB) and total blinks (TB), as well as the partial blink rate (PBR). RESULTS: A total of 1119 children with myopia (2070 eyes) aged 7-18 years were selected. The mean TMH, NIBUT, and LLT of the subjects was 0.21 mm, 12.45 s, and 65.28 nm, respectively. The mean RS and upper and lower MGL scores were 0.64, 1.00, and 1.06, respectively. The mean PB, TB, and PBR was 5.13, 6.46, and 0.81, respectively. Age was significantly correlated to all ocular surface parameters (p = 0.00), except for PB. NIBUT and LLT did not differ between male participants and female participants (all p > 0.05). TMH, RS, and upper and lower MGL were significantly higher in male participants than in female participants (all p < 0.01). In addition, NIBUT was positively associated with TMH (r = 0.13, p = 0.00) and LLT (r = 0.28, p = 0.00). Both upper and lower MGL were positively correlated with TMH, PB, and TB (all p = 0.00), whereas upper MGL was negatively correlated with NIBUT and LLT (all p < 0.05). TB was negatively correlated with NIBUT and LLT (all p = 0.00). PB had no relation with TMH, NIBUT, and LLT (all p > 0.05). In addition, PBR was positively correlated with NIBUT and LLT (all p = 0.00) but not with TMH, RS, or MGL (all p > 0.05). Overall, 57.00% had a TMH ≤ 0.2 mm, 43.20% had a NIBUT ≤ 10 s, 48.10% had an LLT ≤ 60 nm, and 88.10% had a PBR > 0.4. CONCLUSIONS: Child orthokeratology candidates have enhanced tear secretion and increased meibomian gland deficiency with aging. In addition, the adult dry eye diagnostic criteria may apply to orthokeratology candidates aged 12-18 years but should be lower for younger candidates. Given the proportion of abnormal risk, it is necessary to assess tear film status and blink pattern by reliable and feasible objective examination before fitting orthokeratology.

Pinoresinol rescues developmental phenotypes of Arabidopsis phenylpropanoid mutants overexpressing FERULATE 5-HYDROXYLASE.

Most phenylpropanoid pathway flux is directed toward the production of monolignols, but this pathway also generates multiple bioactive metabolites. The monolignols coniferyl and sinapyl alcohol polymerize to form guaiacyl (G) and syringyl (S) units in lignin, components that are characteristic of plant secondary cell walls. Lignin negatively impacts the saccharification potential of lignocellulosic biomass. Although manipulation of its content and composition through genetic engineering has reduced biomass recalcitrance, in some cases, these genetic manipulations lead to impaired growth. The reduced-growth phenotype is often attributed to poor water transport due to xylem collapse in low-lignin mutants, but alternative models suggest that it could be caused by the hyper- or hypoaccumulation of phenylpropanoid intermediates. In Arabidopsis thaliana, overexpression of FERULATE 5-HYDROXYLASE (F5H) shifts the normal G/S lignin ratio to nearly pure S lignin and does not result in substantial changes to plant growth. In contrast, when we overexpressed F5H in the low-lignin mutants cinnamyl dehydrogenase c and d (cadc cadd), cinnamoyl-CoA reductase 1, and reduced epidermal fluorescence 3, plant growth was severely compromised. In addition, cadc cadd plants overexpressing F5H exhibited defects in lateral root development. Exogenous coniferyl alcohol (CA) and its dimeric coupling product, pinoresinol, rescue these phenotypes. These data suggest that mutations in the phenylpropanoid pathway limit the biosynthesis of pinoresinol, and this effect is exacerbated by overexpression of F5H, which further draws down cellular pools of its precursor, CA. Overall, these genetic manipulations appear to restrict the synthesis of pinoresinol or a downstream metabolite that is necessary for plant growth.

Outdoor Scene Classrooms to Arrest Myopia: Design and Baseline Characteristics.

PURPOSE: This study aimed to investigate the impact on childhood myopia of classrooms with spatial properties of classrooms resembling those of outdoor environments. This article describes the design, baseline characteristics, and the acceptability of this strategy. METHODS: Classrooms had custom-made wallpaper installed with forest and sky scenes that had spatial frequency spectra comparable with outdoor environments (i.e., outdoor scene classrooms). Acceptability of this strategy was evaluated by questionnaires. Outcomes to access the efficacy include cumulative proportion of myopia, change of cycloplegic spherical equivalent refractive error, and axial length. RESULTS: Ten classes, comprising 520 students, were randomly assigned into outdoor scene or tradition classrooms. There was no difference in refractive status between two groups (myopia/emmetropia/hyperopia, 16.3% vs. 49.4% vs. 34.2% in outdoor scene classrooms, 18.3% vs. 49.0% vs. 32.7% in traditional classrooms; P = .83). Compared with the traditional classrooms, 88.9% of teachers and 87.5% of students felt the outdoor scene classrooms enjoyable, 22.2% of teachers and 75.3% of students reported higher concentration, and 77.8% of teachers and 15.2% of students reported no change. In addition, 44.4% of teachers and 76.0% of students reported higher learning efficiency in the outdoor scene classrooms, and 55.6% of teachers and 18.3% of students reported no change. CONCLUSIONS: Outdoor scene classrooms are appealing to teachers and students. Outcomes of the study will inform the efficacy of this strategy in Chinese children.

Short-term application of diquafosol ophthalmic solution benefits children with dry eye wearing orthokeratology lens.

Purpose: This aim of this study was to evaluate the effect of 3% Diquafosol Ophthalmic Solution (DQS) on children with dry eye from wearing overnight orthokeratology (OrthoK) lenses. Methods: Myopic children aged 8-18 years with dry eye syndrome were enrolled in this prospective observational study, and they were grouped according to their OrthoK treatment history for at least 1 year. All participants received DQS 4 times per day for 1 month. The following indicators were measured at baseline 1 month after treatment: the Dry Eye Questionnaire-5 (DEQ-5), non-invasive tear meniscus height (TMH), non-invasive tear film break-up time (first and average, NIBUT-F and NIBUT-A), meibomian gland score (MG score), conjunctival hyperemia redness score (R-scan), and blink pattern analysis. Results: A total of 104 participants (189 eyes) including 40 OrthoK wearers (72 eyes) and 64 Orthok candidates (117 eyes) completed the study. Of all, after DQS treatment for 1 month, DEQ-5 scores reduced from 5.54 ± 3.25 to 3.85 ± 2.98 (t = -3.36, p = 0.00). TMH increased from 0.20 ± 0.05 mm to 0.21 ± 0.05 mm (t = 2.59, p = 0.01), NIBUT-F and NIBUT-A were prolonged from 6.67 ± 4.71 s to 10.32 ± 6.19 s and from 8.86 ± 5.25 s to 13.30 ± 6.03 s (all p = 0.00), respectively. R-scan decreased from 0.69 ± 0.28 to 0.50 ± 0.25 (t = -9.01, p = 0.00). Upper MG scores decreased from 1.04 ± 0.32 to 0.97 ± 0.36 (t = -2.14, p = 0.03). Lower MG scores, partial blink rate, partial blinks, and total blinks did not change significantly. Both break-up time (BUT) and R-scan improved significantly after DQS treatment for 1 month (all p = 0.00) in OrthoK candidates and OrthoK wearers. Among the OrthoK wearers, TMH and dry eye symptoms increased significantly (all p = 0.00) but did not increase in OrthoK candidates (p > 0.05). There were no adverse events related to DQS. Conclusion: Diquafosol Ophthalmic Solution was effective for children wearing overnight orthokeratology in relieving dry eye symptoms and improving ocular surface parameters, which may help improve children's OrthoK wearing tolerance and compliance.

Relative Myopic Defocus in the Superior Retina as an Indicator of Myopia Development in Children.

Purpose: To investigate the role of peripheral refraction in children's myopization. Methods: This 2-year study included 214 children (8-15 years old). Refraction across the retina (field of view: 60° × 36°) was measured with a custom-made aberrometer every year. Three datasets were established: dataset 1, 214 subjects from baseline to the first-year visit; dataset 2, 152 subjects from baseline to the second-year visit; and dataset 3, 59 initial emmetropes from baseline to the second-year visit. The baseline refraction of different retina regions was correlated with the central myopic shift, and was compared among groups with different levels of myopic shift. Results: In datasets 1 and 2, the refraction distribution across the retina was significantly different among the subjects who were initially emmetropes but not among those who were initially hyperopic or myopic. Refraction in the central vertical retina, especially in the superior retina (r = -0.5, P < 0.001), was significantly correlated with the myopic shift for emmetropes in that subjects with more relative myopia in the superior retina manifested greater central myopic shifts. In dataset 3, 21 subjects remained emmetropic after 2 years, 15 subjects became myopic at the 1-year visit, and 23 subjects became myopic at the 2-year visit. No difference was found for the relative peripheral refraction in all of the peripheral regions between the stage prior to and after the onset of myopia. Conclusions: Relative myopic defocus in the superior retina could be a predictor of central myopia shift. Changes in relative peripheral refraction are more likely a consequence of myopia progression rather than a cause.

Brain network mechanism on cognitive control task in the elderly with brain aging: A functional near infrared spectroscopy study.

Objective: To study the brain network mechanism of cognitive control in the elderly with brain aging. Materials and methods: 21 normal young people and 20 elderly people were included in this study. Mini-mental State Examination and functional near-infrared spectroscopy (fNIRS) synchronous judgment test (including forward tests and reverse judgment tests) were performed on all subjects. To observe and compare differences in brain region activation and brain functional connectivity between subjects and forward and reverse trials by recording functional connectivity (FC) in different task paradigms and calculating bilateral prefrontal and primary motor cortical (PMC) areas. Results: In the forward and reverse judgment tests, the reaction time of the elderly group was significantly longer than the young group (P < 0.05), and there was no significant difference in the correct rate. In the homologous regions of interest (ROI) data, the FC of PMC and prefrontal cortex (PFC) in the elderly group was significantly decreased (P < 0.05). In the heterologous ROI data, except for left primary motor cortex (LPMC)-left prefrontal cortex (LPFC), the other PMC and PFC of the elderly group were significantly lower than the young group (P < 0.05) while processing the forward judgment test. However, the heterologous ROI data of LPMC-right prefrontal cortex (RPFC), LPMC-LPFC and RPFC-LPFC in the elderly group were significantly lower than the young group (P < 0.05) while processing the reverse judgment test. Conclusion: The results suggest that brain aging affected degeneration of whole brain function, which reduce the speed of information processing and form a brain network functional connection mode different from that of young people.

Multi-disciplinary treatment of severe palatal radicular groove of maxillary lateral incisor: A case report and literature review. / 上颌侧切牙重度畸形舌侧沟多学科联合治疗1ä¾‹å¹¶æ–‡çŒ®å¤ä¹ .

Palatal radicular groove is a developmental malformation of maxillary incisors, lateral incisors in particular, which often causes periodontal destruction. This paper reports a case of combined periodontal-endodontic lesions induced by palatal radicular groove, which was initially misdiagnosed as a simple periapical cyst. After root canal therapy and periapical cyst curettage, the course of disease was prolonged, resulting in the absence of buccal and maxillary bone plates in the affected tooth area. After the etiology was determined, the affected tooth was extracted and guide bone tissue regeneration was performed at the same time, followed by implantation and restoration at the later stage, leading to clinical cure. The palatal radicular groove is highly occult, and the clinical symptoms are not typical. If the abscess of the maxillary lateral incisor occurs repeatedly, and the abscess of the maxillary lateral incisor has not been cured after periodontal and root canal treatment, cone-beam computed tomographic and periodontal flap surgery should be considered.

Solvation Free Energy for Selection of an Aqueous Two-Phase System: Case in Paeonol Extraction from Cortex Moutan.

Aqueous two-phase system(s) (ATPS) has/have been widely employed in the extraction and separation of bioactive molecules from herbs due to its various advantages such as high efficiency and good selectivity. For selecting the type and amount of organics and salts in ATPS, onerous experimental trials are required to ensure the reliability. We intended to develop a theoretical method to select ATPS in the case of paeonol extraction from cortex moutan. The solvation free energies (E solv) of paeonol in the top phase of 54 ATPS (ATPS-acetone, ATPS-acetone-EA, ATPS-THF, ATPS-THF-EA, ATPS-EtOH, and ATPS-EtOH-EA) were calculated with Gaussian 09, and the extraction yields with 54 ATPS were determined. By comparison of E solv and yield, the E solv rank was effective to select the optimal organic type and organic solvent fraction and aqueous salt concentration. With each series of 18 ATPS (ATPS-acetone plus ATPS-acetone-EA; ATPS-THF plus ATPS-THF-EA; or ATPS-EtOH plus ATPS-EtOH-EA), the paeonol yield was correlated with E solv, suggesting that the optimal organic type and fraction and the aqueous NaH2PO4 concentration could be selected by using theoretical E solv, or at least, the theoretical E solv rank could offer effective guidance for experimental design, and thus, tedious and onerous experimental work for optimization in ATPS extraction can be significantly reduced.

Water-carbon nexus relationship and interaction mechanism analysis within Beijing-Tianjin-Hebei urban agglomeration.

As the total water resources consumption control and carbon mitigation continuous improvement, the weak water-carbon incorporate management is increasingly exposed. In this study, a water-carbon nexus assessment framework is proposed to analyze the nexus relationship between water consumption and carbon emission, and distinguishes the coupled water-carbon transmission intensity and the transfer paths under regional and industrial scales. According to the practical input-output table, water consumption, and carbon emission information, the framework is applied to Beijing-Tianjin-Hebei urban agglomeration (BTHUA), a population, resource, and trade intensive area of China. Inter-regional/intra-regional water consumption and carbon emission transfer fluxes between sectors, the pairwise ecological relationship, and the water-carbon nexus were analyzed. Results indicated that the water-carbon transfer indexes from Hebei to Beijing and Tianjin were 161.85 kg/m3 and 113.88 kg/m3 in the study period, along with the most water consumption and carbon emission, and the worst water-carbon nexus. From the industrial perspective, electricity and gas supplying industry provided 7.8% and 29.1% of the total carbon transfer in Tianjin and Hebei, as the most key node sectors on the water-carbon nexus in the BTHUA. The research provides valuably supporting the adjustment of the existing urban agglomeration water-carbon nexus management schemes.

Comparative Analysis of the Microbial Community Structures Between Healthy and Anthracnose-Infected Strawberry Rhizosphere Soils Using Illumina Sequencing Technology in Yunnan Province, Southwest of China.

Anthracnose caused by Colletotrichum spp. was widespread in recent years and resulted in great damage to strawberry production. Soil microbial communities were key contributors to host nutrition, development, and immunity; however, the difference between the microbial communities of healthy and anthracnose-infected strawberry rhizosphere soils remains unclear. In this study, the Illumina sequencing technique was used to comparatively study the prokaryotic and fungal community compositions and structures between healthy and anthracnose-infected strawberry rhizosphere soils in Yuxi, Yunnan Province. Both microbial community diversities and richness of anthracnose-infected strawberry rhizosphere soils were higher than those of healthy strawberry rhizosphere soils. A total of 2,518 prokaryotic and 556 fungal operational taxonomic units (OTUs) were obtained at the 97% similarity threshold. Proteobacteria, Thaumarchaeota, and Acidobacteria were the dominant prokaryotic phyla; Ascomycota, unclassified_k__Fungi, and Mortierellomycota were the dominant fungal phyla. The relative abundances of beneficial bacterial phyla Actinobacteria and Firmicutes, genera Streptomyces, Azospirillum, and Bacillus were significantly reduced in anthracnose-infected strawberry rhizosphere soils; the relative abundance of beneficial fungal species Trichoderma asperellum shows a similar tendency with bacterial abundance. Besides Colletotrichum, 15 other potential fungal pathogen genera and seven fungal pathogen species were identified; among the potential pathogen genera and species, eight pathogen genera and Fusarium oxysporum showed significant differences between healthy and anthracnose-infected strawberry rhizosphere soils. The results suggested that strawberry planted in this area may be infected by other fungal pathogens except for Colletotrichum spp. Our present research will provide theoretical basis and data reference for the isolation and identification of strawberry pathogens and potential probiotics in future works.

Rolling back human pluripotent stem cells to an eight-cell embryo-like stage.

After fertilization, the quiescent zygote experiences a burst of genome activation that initiates a short-lived totipotent state. Understanding the process of totipotency in human cells would have broad applications. However, in contrast to in mice1,2, demonstration of the time of zygotic genome activation or the eight-cell (8C) stage in in vitro cultured human cells has not yet been reported, and the study of embryos is limited by ethical and practical considerations. Here we describe a transgene-free, rapid and controllable method for producing 8C-like cells (8CLCs) from human pluripotent stem cells. Single-cell analysis identified key molecular events and gene networks associated with this conversion. Loss-of-function experiments identified fundamental roles for DPPA3, a master regulator of DNA methylation in oocytes3, and TPRX1, a eutherian totipotent cell homeobox (ETCHbox) family transcription factor that is absent in mice4. DPPA3 induces DNA demethylation throughout the 8CLC conversion process, whereas TPRX1 is a key executor of 8CLC gene networks. We further demonstrate that 8CLCs can produce embryonic and extraembryonic lineages in vitro or in vivo in the form of blastoids5 and complex teratomas. Our approach provides a resource to uncover the molecular process of early human embryogenesis.

Transposon insertions regulate genome-wide allele-specific expression and underpin flower colour variations in apple (Malus spp.).

Allele-specific expression (ASE) can lead to phenotypic diversity and evolution. However, the mechanisms regulating ASE are not well understood, particularly in woody perennial plants. In this study, we investigated ASE genes in the apple cultivar 'Royal Gala' (RG). A high quality chromosome-level genome was assembled using a homozygous tetra-haploid RG plant, derived from anther cultures. Using RNA-sequencing (RNA-seq) data from RG flower and fruit tissues, we identified 2091 ASE genes. Compared with the haploid genome of 'Golden Delicious' (GD), a parent of RG, we distinguished the genomic sequences between the two alleles of 817 ASE genes, and further identified allele-specific presence of a transposable element (TE) in the upstream region of 354 ASE genes. These included MYB110a that encodes a transcription factor regulating anthocyanin biosynthesis. Interestingly, another ASE gene, MYB10 also showed an allele-specific TE insertion and was identified using genome data of other apple cultivars. The presence of the TE insertion in both MYB genes was positively associated with ASE and anthocyanin accumulation in apple petals through analysis of 231 apple accessions, and thus underpins apple flower colour evolution. Our study demonstrated the importance of TEs in regulating ASE on a genome-wide scale and presents a novel method for rapid identification of ASE genes and their regulatory elements in plants.

microRNA172 targets APETALA2 to regulate flavonoid biosynthesis in apple (Malus domestica).

MicroRNA172 (miR172) plays a role in regulating a diverse range of plant developmental processes, including flowering, fruit development and nodulation. However, its role in regulating flavonoid biosynthesis is unclear. In this study, we show that transgenic apple plants over-expressing miR172 show a reduction in red coloration and anthocyanin accumulation in various tissue types. This reduction was consistent with decreased expression of APETALA2 homolog MdAP2_1a (a miR172 target gene), MdMYB10, and targets of MdMYB10, as demonstrated by both RNA-seq and qRT-PCR analyses. The positive role of MdAP2_1a in regulating anthocyanin biosynthesis was supported by the enhanced petal anthocyanin accumulation in transgenic tobacco plants overexpressing MdAP2_1a, and by the reduction in anthocyanin accumulation in apple and cherry fruits transfected with an MdAP2_1a virus-induced-gene-silencing construct. We demonstrated that MdAP2_1a could bind directly to the promoter and protein sequences of MdMYB10 in yeast and tobacco, and enhance MdMYB10 promotor activity. In Arabidopsis, over-expression of miR172 reduced flavonoid (including anthocyanins and flavonols) concentration and RNA transcript abundance of flavonoid genes in plantlets cultured on medium containing 7% sucrose. The anthocyanin content and RNA abundance of anthocyanin genes could be partially restored by using a synonymous mutant of MdAP2_1a, which had lost the miR172 target sequences at mRNA level, but not restored by using a WT MdAP2_1a. These results indicate that miR172 inhibits flavonoid biosynthesis through suppressing the expression of an AP2 transcription factor that positively regulates MdMYB10.