Design Efficacy Evaluation of a Landscape Information Modeling-Stable Diffusion(LIM-SD)-based Approach for Ecological Engineered Landscaping Design:A Case Study of an Urban River Wetland

This study introduces a Landscape Information Modeling±Stable Diffusion(LIM±SD)-based digital workflow for ecological engineered landscaping(EEL)design,focusing on urban river wetlands.It explores how students from diverse academic backgrounds perform EEL tasks using the LIM±SD approach.A total of 30 participants,including industrial design postgraduates and landscape architecture undergraduates and postgraduates,completed the design tasks.The efficacy of their designs was assessed through expert evaluations on site appropriateness,aesthetics,spatial layout,and eco-engineering techniques of the design proposals,as well as the parametric simulation which calculated the vegetation coverage rate and proportion of riparian areas for each design.Moreover,evaluation of participants’subjective design experiences was conducted via questionnaires.Results indicated that landscape architecture postgraduates outperformed others applying ecological engineering principles.The study also elucidated discrepancies between LIM models and SD-generated renderings,as well as the uncertainty of SDgenerated renderings,suggesting improvements are needed to align digital outputs with ecological design criteria.

NIGT1 represses plant growth and mitigates phosphate starvation signaling to balance the growth response tradeoff in rice

Inorganic phosphate(Pi)availability is an important factor which affects the growth and yield of crops,thus an appropriate and effective response to Pi fluctuation is critical.However,how crops orchestrate Pi signaling and growth under Pi starvation conditions to optimize the growth defense tradeoff remains unclear.Here we show that a Pi starvationinduced transcription factor NIGT1(NITRATE-INDUCIBLE GARP-TYPE TRANSCRIPTIONAL REPRESSOR 1)controls plant growth and prevents a hyper-response to Pi starvation by directly repressing the expression of growth-related and Pisignaling genes to achieve a balance between growth and response under a varying Pi environment.NIGT1 directly binds to the promoters of Pi starvation signaling marker genes,like IPS1,mi R827,and SPX2,under Pi-deficient conditions to mitigate the Pi-starvation responsive(PSR).It also directly represses the expression of vacuolar Pi efflux transporter genes VPE1/2 to regulate plant Pi homeostasis.We further demonstrate that NIGT1 constrains shoot growth by repressing the expression of growth-related regulatory genes,including brassinolide signal transduction master regulator BZR1,cell division regulator CYCB1;1,and DNA replication regulator PSF3.Our findings reveal the function of NIGT1 in orchestrating plant growth and Pi starvation signaling,and also provide evidence that NIGT1 acts as a safeguard to avoid hyper-response during Pi starvation stress in rice.

Fargeted elimination of mutant mitochondria DNA in MELAS-iPSCs by mitoTALENs

Mitochondrial diseases are maternally inherited hetero- geneous disorders that are primarily caused by mitochondrial DNA （mtDNA） mutations. Depending on the ratio of mutant to wild-type mtDNA, known as heteroplasmy, mitochondrial defects can result in a wide spectrum of clinical manifestations. Mitochondria-targeted endonucleases provide an alternative avenue for treating mitochondrial disorders via targeted destruc- tion of the mutant mtDNA and induction of heteroplasmic shifting. Here, we generated mitochondrial disease patient-specific induced pluripotent stem cells （MiPSCs） that harbored a high proportion of m.3243A〉G mtDNA mutations and caused mitochondrial encephalomyopathy and stroke-like episodes （MELAS）. We engineered mitochondrial-targeted transcription activator-like effector nucleases （mitoTALENs） and successfully eliminated the m.3243A〉G mutation in MiPSCs. Off-target mutagenesis was not detected in the targeted MiPSC clones. Utilizing a dual fluorescence iPSC reporter cell line expressing a 3243G mutant mtDNA sequence in the nuclear genome, mitoTALENs displayed a significantly limited ability to target the nuclear genome compared with nuclear-localized TALENs. Moreover, genetically rescued MiPSCs displayed normal mitochondrial respiration and energy production. Moreover, neuronal progenitor cells differentiated from the rescued MiPSCs also demonstrated normal metabolic profiles. Further- more, we successfully achieved reduction in the human m.3243A〉G mtDNA mutation in porcine oocytes via injection of mitoTALEN mRNA. Our study shows the great potential for using mitoTALENs for specific targeting of mutant mtDNA both in iPSCs and mammalian oocytes, which not only provides a new avenue for studying mitochondrial biology and disease but also suggests a potential therapeutic approach for the treatment of mitochondrial disease, as well as the prevention of germline transmission of mutant mtDNA.

Controlling the strontium-doping in calcium phosphate microcapsules through yeastregulated biomimetic mineralization

Yeast cells have controllable biosorption on metallic ions during metabolism.However,few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in calcium phosphate microcapsules.In this study,the yeast cells were allowed to pre-adsorb strontium ions metabolically and then served as sacrificing template for the precipitation and calcination of mineral shell.The pre-adsorption enabled the microorganism to enrich of strontium ions into the inner part of the microcapsules,which ensured a slow-release profile of the trace element from the microcapsule.The co-culture with human marrow stromal cells showed that gene expressions of alkaline phosphatase and Collagen-I were promoted.The promotion of osteogenic differentiation was further confirmed in the 3D culture of cell-material complexes.The strategy using living microorganism as‘smart doping apparatus’to control incorporation of trace element into calcium phosphate paved a pathway to new functional materials for hard tissue regeneration.

Mediating Mesenchymal Stem Cells Responses and Osteopontin Adsorption via Oligo(ethylene glycol)-amino Mixed Self-assembled Monolayers

Oligo（ethylene glycol） （-OEG） and amino （-NH2） mixed self-assembled monolayers （SAMs） were employed as model substrates to investigate the effect of charge density on the fate of mesenchymal stem cells （MSCs） and osteopontin （OPN） adsorption. We found that all surfaces presenting -NH2 groups favored cell responses regardless of the surface charge. Meanwhile, OPN adsorption could remain stable on the mixed SAMs over a certain range of charge densities. Our work provides some insights into cell responses and protein adsorption to surface charge.

Exploring visitors’visual perception along the spatial sequence in temple heritage spaces by quantitative GIS methods:a case study of the Daming Temple,Yangzhou City,China

The Daming Temple,built during 457–464 C.E.,is one of the developing ancient temple heritage spaces located in Yangzhou city,P.R.China.Over the past 60 years,variation in visitors’spatial perception along the tour routes in the temple has occurred.This research attempts to reveal the changes in visitors’visual perception along the spatial sequences at 3 different times(i.e.,1962,1973 and 2022).A quantitative GIS-based method,which includes analysing the distribution of visitors’spatial preferences and spatial configuration,is proposed.Digital landscape tools and quantitative estimation methods are used,including mapping within Rhinoceros software,the kernel density estimation(KDE)method within ArcGIS software and spatial syntax analysis within DepthMap software.Extracted geodata from 500 photographs of the heritage space taken by volunteer visitors are analysed within the GIS environment.Values of the mean depth(MD)at both levels of visibility and accessibility are calculated within the visibility graph analysis(VGA)model.Comparisons between the visual preferences of the visitors and the spatial configuration along the spatial sequence are conducted.The results indicate that the spatial sequence has a significant impact on visitors’visual preferences and tour routes.The phenomenon of spatial sequence among dynamic temporal variations and the effects of narrative spaces along the spatial sequence are highlighted and explained,which reveal the relationship between visitors’geospatial preference and the spatial configuration of the temple.Some suggestions are put forwards for further studies on the revitalisation and management of East Asian ancient temple heritage spaces.

Insight into vitronectin structural evolution on material surface chemistries: The mediation for cell adhesion

Biomaterial surface chemistry engenders profound consequences on cell adhesion and the ultimate tissue response by adsorbing proteins from extracellular matrix,where vitronectin(Vn)is involved as one of the crucial mediator proteins.Deciphering the adsorption behaviors of Vn in molecular scale provides a useful account of how to design biomaterial surfaces.But the details of structural dynamics and consequential biological effect remain elusive.Herein,both experimental and computational approaches were applied to delineate the conformational and orientational evolution of Vn during adsorption onto self-assembled monolayers(SAMs)terminating with-COOH,-NH2,-CH3 and-OH.To unravel the interplay between cell binding and the charge and wettability of material surface,somatomedin-B(SMB)domain of Vn holding the RGD cell-binding motif was employed in molecular dynamics(MD)simulations,with orientation initialized by Monte Carlo(MC)method.Experimental evidences including protein adsorption,cell adhesion and integrin gene expressions were thoroughly investigated.The adsorption of Vn on different surface chemistries showed very complex profiles.Cell adhesion was enabled on all Vn-adsorbed surfaces but with distinct mechanisms mostly determined by conformational change induced reorientation.Higher amount of Vn was observed on negatively charged surface(COOH)and hydrophobic surface(CH3).However,advantageous orientations defined by RGD loop conditions were only obtained on the charged surfaces(COOH and NH2).Specifically,COOH surface straightened up the Vn molecules and accumulated them into a higher density,whereas CH3 surface squashed Vn and stacked them into higher density multilayer by tracking adsorption but with the RGD loops restrained.These findings may have a broad implication on the understanding of Vn functionality and would help develop new strategies for designing advanced biomaterials.

Mechanistic insights into the adsorption and bioactivity of fibronectin on surfaces with varying chemistries by a combination of experimental strategies and molecular simulations

Fibronectin(Fn)is significant to the performance of biomaterials,and the chemistry of biomaterial surface play important roles in Fn adsorption and subsequent cell behavior.However,the“molecular scale”mechanism is still unclear.Herein,we combined experimental strategies with molecular simulations to solve this problem.We prepared self-assembled monolayers with varying chemistries,i.e.,SAMs-CH3,SAMs-NH2,SAMs-COOH and SAMs-OH,and characterized Fn adsorption and cell behaviors on them.Next,Monte Carlo method and all-atom molecular dynamics simulations were employed to reveal the orientation/conformation of Fn on surfaces.We found that SAMs-CH3 strongly adsorbed Fn via hydrophobic interactions,but show poor bioactivity as the low exposure of RGD/PHSRN motifs and the deformation of Fn.SAMs-NH2 and SAMs-COOH could adsorb Fn efficiently via vdW interactions,electrostatic interactions,hydrogen bonds and salt bridges.Fn exhibited excellent bioactivity for cell adhesion,proliferation and osteogenic differentiation as high exposure of bioactive motifs on SAMs-NH2,or as the activation of other inferior cell-binding motifs on SAMs-COOH.SAMs-OH showed poor Fn adsorption as the water film.However,the adsorbed Fn displayed non-negligible bioactivity due to high exposure of PHSRN motif and large degree of protein flexibility.We believe that the revealed mechanism presents great potential to rationally design Fn-activating biomaterials.