Effects of freeze-thaw cycle and dry-wet alternation on slope stability

The typical loess on high slopes along the BaoLan High-speed Rail, China, was selected as the research object. The influence of the freeze-thaw cycle and dry-wet alternation on the shear-strength parameters of the unsaturated loess was investigated by laboratory experimental methods. Moreover, the temperature field, seepage field, and stability of slopes with different gradients were simulated under the effect of the freeze-thaw cycle and dry-wet alternation by using the geotechnical analysis software Geo-Studio. The research results showed(1) when the freeze-thaw cycle was repeated on the slope, with the frozen depth increasing, the melted depth did the same; besides, the closed loop of isotherms formed on the slope;(2) under the action of dry-wet circulation, the negative pore-water pressure and volumetric water content showed an upward tendency. However, owing to the different slope gradients, rainfall infiltration was not the same. As time went by, the differences of the negative pore-water pressure and volumetric water content between the slopes of different gradients continued to increase;(3) with the freeze-thaw cycle and dry-wet alternation increasing, the slope-safety factor decreased. Especially in the early period, the slope-safety factor changed remarkably. For slopes undergoing freeze-thaw action, the slope-safety factor was negatively correlated with the gradient. However, with regard to slopes undergoing dry-wet alternation, the result became more complex because the slope-safety factor was related to both seepage strength and slope grade. Accordingly, further research is needed to study the effect of seepage strength and slope grade on the stability of loess slopes.

Alternation and Evolution of the Fables of Pre-Qin Scholars Along With the Contextual Changes

The fables of Pre-Qin scholars occupied an important position in the history of the development of Chinese culture,thoughts,and literature,hence had profound and extensive influence on later generations.As a kind of carrier of thoughts,the fables of the Pre-Qin scholars were not immutable and frozen,for they have been influenced by various aspects in the process of transmitting and recognizing.This article attempts to study the evolution of the function,expression,and connotation of the fables of the Pre-Qin scholars.

Exploring the mechanical behavior and microstructure of compacted loess subjected to dry-wet cycles and chemical contamination

Due to climatic factors and rapid urbanization,the soil in the Loess Plateau,China,experiences the coupled effects of dry-wet cycles and chemical contamination.Understanding the mechanical behavior and corresponding microstructural evolution of contaminated loess subjected to dry-wet cycles is essential to elucidate the soil degradation mechanism.Therefore,direct shear and consolidation tests were performed to investigate the variations in mechanical properties of compacted loess contaminated with acetic acid,sodium hydroxide,and sodium sulfate during dry-wet cycles.The mechanical response mechanisms were investigated using zeta potential,mineral chemical composition,and scanning electron microscopy(SEM)tests.The results indicate that the mechanical deterioration of sodium hydroxidecontaminated loess during dry-wet cycles decreases with increasing contaminant concentration,which is mainly attributed to the thickening of the electrical double layer(EDL)by Nat and the precipitation of calcite,as well as the formation of colloidal flocs induced by OH,thus inhibiting the development of large pores during the dry-wet process.In contrast,the attenuation of mechanical properties of both acetic acid-and sodium sulfate-contaminated loess becomes more severe with increasing contaminant concentration,with the latter being more particularly significant.This is primarily due to the reduction of the EDL thickness and the erosion of cement in the acidic environment,which facilitates the connectivity of pores during dry-wet cycles.Furthermore,the salt expansion generated by the drying process of saline loess further intensifies the structural disturbance.Consequently,the mechanical performance of compacted loess is sensitive to both pollutant type and concentration,exhibiting different response patterns in the dry-wet cycling condition.

Effects of dry-wet cycles on the mechanical properties of sandstone with unloading-induced damage

Sandstone is the fundamental material in various engineering and construction projects.However,the mechanical integrity of sandstone can be compromised by initial unloading damage resulting from activities such as engineering excavations.Furthermore,this degradation is further exacerbated under periodic dry-wet environmental conditions.This study investigated the effects of dry-wet cycles and unloading on the mechanical properties of jointed fine sandstone using uniaxial and triaxial compression tests.These tests were performed on rock samples subjected to varying unloading degrees and different numbers of dry-wet cycles.The results demonstrate that with an increase in the unloading degree from 0%to 70%,there is a corresponding decrease in peak stress ranging from 10%to 33%.Additionally,the cohesion exhibits a reduction of approximately 20%to 25%,while the internal friction angle experiences a decline of about 3.5%to 6%.These findings emphasize a significant unloading effect.Moreover,the degree of peak stress degradation in unloading jointed fine sandstone diminishes with an increase in confining pressure,suggesting that confining pressure mitigates the deterioration caused by dry-wet cycles.Additionally,as the number of dry-wet cycles increases,there is a notable decline in the mechanical properties of the sandstone,evidencing significant dry-wet degradation.Utilizing the Drucker Prager criterion,this study establishes a strength criterion and fracture criterion,denoted as σ_(1)(m,n) and K_(T)^(Ⅱ)(m, n), to quantify the combined impacts of dry-wet cycles and unloading on jointed fine sandstone,which provides a comprehensive understanding of its mechanical behavior under such conditions.

Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

Alternate cotton-peanut intercropping:a new approach to increasing productivity and minimizing environmental impact

Recent publications have highlighted the development of an alternate cotton-peanut intercropping as a novel strat-egy to enhance agricultural productivity.In this article,we provide an overview of the progress made in the alternate cotton-peanut intercropping,specifically focusing on its yield benefits,environmental impacts,and the underlying mechanisms.In addition,we advocate for future investigations into the selection or development of appropriate crop varieties and agricultural equipment,pest management options,and the mechanisms of root-canopy interactions.This review is intended to provide a valuable reference for understanding and adopting an alternate intercropping system for sustainable cotton production.

Pile foundation in alternate layered liquefiable and non-liquefiable soil deposits subjected to earthquake loading

Pile foundations are still the preferred foundation system for high-rise structures in earthquake-prone regions.Pile foundations have experienced failures in past earthquakes due to liquefaction.Research on pile foundations in liquefiable soils has primarily focused on the pile foundation behavior in two or three-layered soil profiles.However,in natural occurrence,it may occur in alternative layers of liquefiable and non-liquefiable soil.However,the experimental and/or numerical studies on the layered effect on pile foundations have not been widely addressed in the literature.Most of the design codes across the world do not explicitly mention the effect of sandwiched non-liquefiable soil layers on the pile response.In the present study,the behavior of an end-bearing pile in layered liquefiable and non-liquefiable soil deposit is studied numerically.This study found that the kinematic bending moment is higher and governs the design when the effect of the sandwiched non-liquefied layer is considered in the analysis as opposed to when its effect is ignored.Therefore,ignoring the effect of the sandwiched non-liquefied layer in a liquefiable soil deposit might be a nonconservative design approach.

Effect of dry-wet cycles on dynamic properties and microstructures of sandstone:Experiments and modelling

Underground pumped storage power plant(UPSP)is an innovative concept for space recycling of abandoned mines.Its realization requires better understanding of the dynamic performance and durability of reservoir rock.This paper conducted ultrasonic detection,split Hopkinson pressure bar(SHPB)impact,mercury intrusion porosimetry(MIP),and backscatter electron observation(BSE)tests to investigate the dynamical behaviour and microstructure of sandstone with cyclical dry-wet damage.A coupling FEM-DEM model was constructed for reappearing mesoscopic structure damage.The results show that dry-wet cycles decrease the dynamic compressive strength(DCS)with a maximum reduction of 39.40%,the elastic limit strength is reduced from 41.75 to 25.62 MPa.The sieved fragments obtain the highest crack growth rate during the 23rd dry-wet cycle with a predictable life of 25 cycles for each rock particle.The pore fractal features of the macropores and micro-meso pores show great differences between the early and late cycles,which verifies the computational statistics analysis of particle deterioration.The numerical results show that the failure patterns are governed by the strain in pre-peak stage and the shear cracks are dominant.The dry-wet cycles reduce the energy transfer efficiency and lead to the discretization of force chain and crack fields.

Alternating current heating techniques for lithium-ion batteries in electric vehicles:Recent advances and perspectives

The significant decrease in battery performance at low temperatures is one of the critical challenges that electric vehicles(EVs)face,thereby affecting the penetration rate in cold regions.Alternating current(AC)heating has attracted widespread attention due to its low energy consumption and uniform heating advantages.This paper introduces the recent advances in AC heating from the perspective of practical EV applications.First,the performance degradation of EVs in low-temperature environments is introduced briefly.The concept of AC heating and its research methods are provided.Then,the effects of various AC heating methods on battery heating performance are reviewed.Based on existing studies,the main factors that affect AC heating performance are analyzed.Moreover,various heating circuits based on EVs are categorized,and their cost,size,complexity,efficiency,reliability,and heating rate are elaborated and compared.The evolution of AC heaters is presented,and the heaters used in brand vehicles are sorted out.Finally,the perspectives and challenges of AC heating are discussed.This paper can guide the selection of heater implementation methods and the optimization of heating effects for future EV applications.

A Neural-network-based Alternative Scheme to Include Nonhydrostatic Processes in an Atmospheric Dynamical Core

Here,a nonhydrostatic alternative scheme(NAS)is proposed for the grey zone where the nonhydrostatic impact on the atmosphere is evident but not large enough to justify the necessity to include an implicit nonhydrostatic solver in an atmospheric dynamical core.The NAS is designed to replace this solver,which can be incorporated into any hydrostatic models so that existing well-developed hydrostatic models can effectively serve for a longer time.Recent advances in machine learning(ML)provide a potential tool for capturing the main complicated nonlinear-nonhydrostatic relationship.In this study,an ML approach called a neural network(NN)was adopted to select leading input features and develop the NAS.The NNs were trained and evaluated with 12-day simulation results of dry baroclinic-wave tests by the Weather Research and Forecasting(WRF)model.The forward time difference of the nonhydrostatic tendency was used as the target variable,and the five selected features were the nonhydrostatic tendency at the last time step,and four hydrostatic variables at the current step including geopotential height,pressure in two different forms,and potential temperature,respectively.Finally,a practical NAS was developed with these features and trained layer by layer at a 20-km horizontal resolution,which can accurately reproduce the temporal variation and vertical distribution of the nonhydrostatic tendency.Corrected by the NN-based NAS,the improved hydrostatic solver at different horizontal resolutions can run stably for at least one month and effectively reduce most of the nonhydrostatic errors in terms of system bias,anomaly root-mean-square error,and the error of the wave spatial pattern,which proves the feasibility and superiority of this scheme.

Improved genome annotation of Brassica oleracea highlights the importance of alternative splicing

Brassica oleracea has been developed into many important crops,including cabbage,kale,cauliflower,broccoli and so on.The genome and gene annotation of cabbage(cultivar JZS),a representative morphotype of B.oleracea,has been widely used as a common reference in biological research.Although its genome assembly has been updated twice,the current gene annotation still lacks information on untranslated regions(UTRs)and alternative splicing(AS).Here,we constructed a high-quality gene annotation(JZSv3)using a full-length transcriptome acquired by nanopore sequencing,yielding a total of 59452 genes and 75684 transcripts.Additionally,we re-analyzed the previously reported transcriptome data related to the development of different tissues and cold response using JZSv3 as a reference,and found that 3843 out of 11908 differentially expressed genes(DEGs)underwent AS during the development of different tissues and 309 out of 903 cold-related genes underwent AS in response to cold stress.Meanwhile,we also identified many AS genes,including BolLHCB5 and BolHSP70,that displayed distinct expression patterns within variant transcripts of the same gene,highlighting the importance of JZSv3 as a pivotal reference for AS analysis.Overall,JZSv3 provides a valuable resource for exploring gene function,especially for obtaining a deeper understanding of AS regulation mechanisms.

Wetting alternating with partial drying during grain filling increases lysine biosynthesis in inferior rice grain

Lysine content is a criterion of the nutritional quality of rice.Understanding the process of lysine biosynthesis in early-flowering superior grain(SG)and late-flowering inferior grain(IG)of rice would advance breeding and cultivation to improve nutritional quality.However,little information is available on differences in lysine anabolism between SG and IG and the underlying mechanism,and whether and how irrigation regimes affect lysine anabolism in these grains.A japonica rice cultivar was grown in the field and two irrigation regimes,continuous flooding(CF)and wetting alternating with partial drying(WAPD),were imposed from heading to the mature stage.Lysine content and activities of key enzymes of lysine biosynthesis,and levels of brassinosteroids(BRs)were lower in the IG than in the SG at the early grainfilling stage but higher at middle and late grain-filling stages.WAPD increased activities of these key enzymes,BR levels,and contents of lysine and total amino acids in IG,but not SG relative to CF.Application of 2,4-epibrassinolide to rice panicles in CF during early grain filling reproduced the effects of WAPD,but neither treatment altered the activities of enzymes responsible for lysine catabolism in either SG or IG.WAPD and elevated BR levels during grain filling increased lysine biosynthesis in IG.Improvement in lysine biosynthesis in rice should focus on IG.

2,3,5,4’-Tetrahydroxystilbene-2-O-b-D-Glucoside modulates CHEK2 and CCND1 alternative splicing to inhibit MCF-7 cells proliferation

Background:In our previous study,we observed a synergistic effect of 2,3,5,4’-Tetrahydroxystilbene-2-O-b-D-glucoside combined with adriamycin to induce apoptosis in MCF-7 breast cancer cells.However,the underlying mechanisms of epigenetic modifications,such as alternative splicing,have not been explored.In this study,we aimed to investigate the mechanism by which THSG inhibits MCF-7 cell proliferation using full-length transcriptome sequencing.Methods:First,cell viability was examined using the methyl thiazolyl tetrazolium method and full-length transcriptome sequencing was performed to identify genes and pathways.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes were used to identify the principal pathways and targets of THSG.Flow cytometry analysis of cell cycle distribution was performed.Meanwhile,the analysis of alternative splicing and domains of the key proteins was conducted.Quantitative polymerase chain reaction and western blotting were performed for verification.Results:THSG showed significant cytotoxic activity in MCF-7 cells.Full-length transcriptome sequencing revealed differential alternative splicing with 173 upregulated and 263 downregulated genes.Further analysis identified distinct differential expression of genes(CHEK2-211 and CCND1-201)involved in the cell cycle in the THSG-treated group.Subsequently,alternative splicing types of CHEK2(mutually exclusive exon)and CCND1(intron retention).We found that THSG downregulated mRNA expression,as confirmed by quantitative polymerase chain reaction analysis.Interestingly,protein structural analysis revealed that THSG treatment led to the generation of CHK2-211,which was the result of a mutation in the amino acid residues(GLU-150,ASN-151)of the CHEK2 domain(VAL-150,GLY-151).and CyclinD1-201 were obtained when an amino acid(ASP-267)in the domain was lost in CyclinD1.Moreover,molecular docking analysis demonstrated that the domains of key proteins could bind THSG more effectively,with no difference in affinity.Western blotting confirmed that THSG inhibited the expression of CHK2 and CyclinD1.Conclusion:THSG modulated the alternative splicing of CHEK2 and CCND1 by inducing G0/G1 cell cycle arrest,consequently suppressing MCF-7 cell proliferation.

Suppression of stimulated Brillouin and Raman scatterings using an alternating frequency laser and transverse magnetic fields

A novel scheme to suppress both stimulated Brillouin scattering(SBS) and stimulated Raman scattering(SRS) by combining an alternating frequency(AF) laser and a transverse magnetic field is proposed. The AF laser allows the laser frequency to change discretely and alternately over time. The suppression of SBS is significant as long as the AF difference is greater than the linear growth rate of SBS or the alternating time of the laser frequency is shorter than the linear growth time of SBS. However, the AF laser proves ineffective in suppressing SRS, which usually has a much higher linear growth rate than SBS. To remedy that, a transverse magnetic field is included to suppress the SRS instability. The electrons trapped in the electron plasma waves(EPWs) of SRS can be accelerated by the surfatron mechanism in a transverse magnetic field and eventually detrapped. While continuously extracting energy from EPWs, the EPWs are dissipated and the kinetic inflation of SRS is suppressed. The one-dimensional particle-in-cell simulation results show that both SBS and SRS can be effectively suppressed by combining the AF laser with a transverse magnetic field with tens of Tesla. The total reflectivity can be dramatically reduced by more than one order of magnitude. These results provide a potential reference for controlling SBS and SRS under the related parameters of inertial confinement fusion.

Workshop “Navigating the Transition: Standards Powering the Journey of Alternative Fuel Infrastructure”

April 18, Brussels, Belgium In light of recent significant regulatory changes within the European Union impacting the transport sector, such as the Alternative Fuels Infrastructure Regulation(AFIR), Re Fuel EU and Fuel EU Maritime, the role of European standardization becomes increasingly critical in facilitating the transition towards the deployment of alternative fuels.

Diagnosing ratio of electron density to collision frequency of plasma surrounding scaled model in a shock tube using low-frequency alternating magnetic field phase shift

A non-contact low-frequency(LF)method of diagnosing the plasma surrounding a scaled model in a shock tube is proposed.This method utilizes the phase shift occurring after the transmission of an LF alternating magnetic field through the plasma to directly measure the ratio of the plasma loop average electron density to collision frequency.An equivalent circuit model is used to analyze the relationship of the phase shift of the magnetic field component of LF electromagnetic waves with the plasma electron density and collision frequency.The applicable range of the LF method on a given plasma scale is analyzed.The upper diagnostic limit for the ratio of the electron density(unit:m^(-3))to collision frequency(unit:Hz)exceeds 1×10^(11),enabling an electron density to exceed 1×10^(20)m^(-3)and a collision frequency to be less than 1 GHz.In this work,the feasibility of using the LF phase shift to implement the plasma diagnosis is also assessed.Diagnosis experiments on shock tube equipment are conducted by using both the electrostatic probe method and LF method.By comparing the diagnostic results of the two methods,the inversion results are relatively consistent with each other,thereby preliminarily verifying the feasibility of the LF method.The ratio of the electron density to the collision frequency has a relatively uniform distribution during the plasma stabilization.The LF diagnostic path is a loop around the model,which is suitable for diagnosing the plasma that surrounds the model.Finally,the causes of diagnostic discrepancy between the two methods are analyzed.The proposed method provides a new avenue for diagnosing high-density enveloping plasma.

Use of Topical Complementary and Alternative Medicines (CAM) in Diabetic Septic Foot Disease in Makkah Region, Western of Saudi Arabia, an Exploratory Study

Diabetes is a chronic condition that is significantly impacted daily by environment. At least one in five Saudis, suffer from diabetes. “Diabetes is a pandemic of unprecedented magnitude spiraling out of control” (Boulton). Saudi Arabia ranks the second highest in the Middle East, and is the seventh in the world for the prevalence of diabetes (WHO). Diabetes mellitus (DM) has been found to be related to high mortality, morbidity accompanied by poor general health and lower quality of life. Current Saudi diabetes patients’ behaviors, practices, and beliefs regarding foot health problems such as ulcers, are not well investigated. Diabetic patients frequently utilize natural remedies and Herbs for self-medication as a part of complementary and alternative Medicine. Aim of study: To explore the widespread use of the alternative remedies in the local communities of the Western Saudi Arabia, the drives behind using and to focus on the potential adverse effects following their usage. Method: A descriptive, cross-sectional study of 386 diabetic patients with feet problems in Makkah Region, West of Saudi Arabia during the period April - June 2023. Results: Out of the 531 participants, only 386 participants reported diabetic foot disorder (DFD). Most of the participant were above 65 years of age group (24.61 %). Male (52.08%) more than female (47.92%) in the research group. All of the participants had diabetic foot disorders at least once. The prevalence of complementary and alternative medicines (CAM) use among them was 67.88%, as solitary or in combination with hospital medicines. Honey headed the list followed by myrrh and black seeds. Acquaintances advice (59.54%) was the most reason for using the alternative medicine. 60.30% of Diabetic foot patient complained of adverse effects during CAM remedies use. Conclusion: Consumption of CAM remedies among diabetic patients for Diabetic Foot Disorder (DFD) is common. Honey headed the list followed by myrrh and black seeds. Additional wide-scale research is required to establish their actual efficacy, safety and potential adverse out-comes and to generalize the outcomes.

Molecular targets and mechanisms of different aberrant alternative splicing in metastatic liver cancer

Metastasis remains a major challenge in the successful management of malignant diseases.The liver is a major site of metastatic disease and a leading cause of death from gastrointestinal malignancies such as colon,stomach,and pancreatic cancers,as well as melanoma,breast cancer,and sarcoma.As an important factor that influences the development of metastatic liver cancer,alternative splicing drives the diversity of RNA transcripts and protein subtypes,which may provide potential to broaden the target space.In particular,the dysfunction of splicing factors and abnormal expression of splicing variants are associated with the occurrence,progression,aggressiveness,and drug resistance of cancers caused by the selective splicing of specific genes.This review is the first to provide a detailed summary of the normal splicing process and alterations that occur during metastatic liver cancer.It will cover the role of alternative splicing in the mechanisms of metastatic liver cancer by examining splicing factor changes,abnormal splicing,and the contribution of hypoxia to these changes during metastasis.

甘肃省陇南市核桃枝枯病菌Alternaria alternate的生物学特性

以核桃枝枯病菌(Alternaria alternate)为对象,研究温度、碳源、氮源、光照、培养基和pH等对菌丝生长、产孢量和孢子萌发的影响。结果表明,pH 7和全黑暗有利于核桃枝枯病菌的菌丝生长、产孢和孢子萌发;25℃有利于菌丝生长,25~30℃有利于产孢,20~30℃有利于孢子萌发;最适生长的碳源为可溶性淀粉,氮源为酵母浸粉、KNO_(3)和蛋白胨,最适产孢的碳源和氮源分别为山梨醇和KNO_(3),最适孢子萌发的碳源和氮源分别为乳糖和NH_(4)NO_(3);PSA、OA和PDA培养基适合菌丝生长,LB培养基适合产孢,OA培养基适合孢子萌发。该研究结果为明确核桃枝枯病的发生规律提供理论依据,也为综合防治措施的制定提供参考。

Progression of Corneal Astigmatism after Phaco-Alternative at the University Hospital Center of the Oueme-Plateau Department in Porto-Novo

Objective: To study the postoperative evolution of corneal astigmatism following phaco-alternative at the University Hospital Center of Ouémé-Plateau (UHC-OP) in Porto-Novo. Method: This was a prospective, cross-sectional study with a descriptive and analytical purpose conducted from April 19 to September 20, 2021;a period of 6 months. The study included all patients presenting with senile or non-senile cataracts without a history of corneal trauma and who underwent phaco-alternative during the study period. Results: A total of 62 eyes underwent phaco-alternative. The average age was 63 ± 12 years with a male predominance of 59.7%. Phaco-alternative was performed with 74.2% linear incision and 25.8% smile incision. Perioperative incidents were mainly capsular ruptures with vitreous loss occurring in 8.1% of cases. The mean preoperative astigmatism was 1.29 ± 1.5 D with an axis of 180˚ ± 20˚, indicating with-the-rule astigmatism. The mean postoperative astigmatism was 2.2 ± 1.5 D with an axis of 90˚ ± 20˚, indicating against-the-rule astigmatism. The mean induced astigmatism at Day 30 was 1.4 ± 1.2 D for smile incision and 1.8 ± 1.2 D for linear incision. Conclusion: Phaco-alternative yields good results with few complications but remains astigmatogenic. The smile incision appears to be less astigmatogenic.