Commercial Products Research Articles

Optimizing Surface Texture Through Ion Induction for High Areal Capacitance and Enhanced Stability in MXene Electrodes

AbstractTo make supercapacitors a viable commercial product, it is necessary to increase the mass loading (ML) of the electrodes. However, current research on high ML MXene electrodes mainly focuses on internal structure optimization, often neglecting the importance of surface texture in the electrodes, which plays a crucial role in mediating interactions between the internal structure and the electrolyte. This study reports on a simple ion‐intercalation process that can reduce charge transfer resistance and improve cycling stability. Through the integration of 3D visualization models and wide‐angle X‐ray scattering techniques, a comprehensive investigation of the electrode's surface structure and MXene flake arrangement is conducted. The results showed that pre‐oriented aggregates induced by ion‐intercalation create a sophisticated surface structure with rich hills and valleys, promoting electrolyte permeation and ion exchange. This work highlights the significance of surface texture in films and electrodes, guiding the design of high‐performance materials.

Laboratory performances during proficiency tests for the analysis of active substances in pesticide formulations during 2021–2023

During the period 2021–2023, the Italian National Institute of Health (ISS) organised, in collaboration with the Italian Health Ministry, proficiency testing (PTs) for about 22 Italian and other EU laboratories involved in the monitoring programmes for Plant Protection Products (PPPs). The aim of the PTs was to determine the amount of active ingredient in various formulations of PPPs. Overall, the type of formulations to be analysed were soluble concentrate (SL), suspension concentrate (SC), capsule suspension (CS), emulsion water oil (EW), soluble water granule (SG), dustable powder (DP), water dispersible granule (WDG) and emulsion concentrate (EC). In 2021, the ISS organised PTs for the determination of Imazamox, Fenhexamid and Trinexapac-ethyl in three different PPPs as soluble concentrate (SL), suspension concentrate (SC), and capsule suspension (CS), respectively. In 2022, PTs for the determination of Bentazone, Spinosad and Tau-Fluvalinate were organised. For this PTs, commercial PPPs were obtained from the Italian market as SC, emulsion oil water (EW) and soluble water granule (SG). In 2023, the PT was organised to determine the active ingredient Trifloxystrobin, Deltamethrin and Cyprodinil in the commercial product as water dispersible granule (WDG), dustable powder (DP) and emulsion concentrate (EC), respectively. Homogeneity and stability, performed for all PT samples, fulfilled the acceptability criteria. The Jarque–Bera test was used to verify the hypothesis of normality of the data distributions before to calculate the z-score values. Laboratory performances, expressed in terms of z-scores, were satisfactory for almost all participants for all substances.

Recent Progress in Terrestrial Biota Derived Antibacterial Agents for Medical Applications

Conventional antibiotic and multidrug treatments are becoming less and less effective and the discovery of new effective and safe antibacterial agents is becoming a global priority. Returning to a natural antibacterial product is a relatively new current trend. Terrestrial biota is a rich source of biologically active substances whose antibacterial potential has not been fully utilized. The aim of this review is to present the current state-of-the-art terrestrial biota-derived antibacterial agents inspired by natural treatments. It summarizes the most important sources and newly identified or modified antibacterial agents and treatments from the last five years. It focuses on the significance of plant- animal- and bacteria-derived biologically active agents as powerful alternatives to antibiotics, as well as the advantages of utilizing natural antibacterial molecules alone or in combination with antibiotics. The main conclusion is that terrestrial biota-derived antibacterial products and substances open a variety of new ways for modern improved therapeutic strategies. New terrestrial sources of known antibacterial agents and new antibacterial agents from terrestrial biota were discovered during the last 5 years, which are under investigation together with some long-ago known but now experiencing their renaissance for the development of new medical treatments. The use of natural antibacterial peptides as well as combinational therapy by commercial antibiotics and natural products is outlined as the most promising method for treating bacterial infections. In vivo testing and clinical trials are necessary to reach clinical application.

Marker trait association and candidate gene identification for brown rust disease in sugarcane

AbstractBrown rust (caused by Puccinia melanocephala H. & P. Sydow) is one of the most devastating diseases in commercial sugarcane production. It could reduce sugarcane yield by up to 50% depending on the susceptibility levels of cultivars. Breeding disease‐resistant cultivars is the most effective, economical, and environmentally friendly option to control brown rust. A genome‐wide association study was conducted on a field trial using 432 sugarcane clones following an augmented design with two replications. Brown rust was screened using the whorl inoculation method over two crop cycles. The genotype data were obtained through target enrichment sequencing technologies. The gene actions considering six different models and marker dosage effects were included during the marker‐trait analysis. A total of seven, nine, and seven nonredundant marker‐trait associations were identified for plant cane, first ratoon, and across two crop cycles, respectively. The most significant (p‐value 6.17E−20) marker (chr01p59833543) has the additive effect of −0.63 for the diplo‐additive model and reduced disease severity the most (41.35%) due to heterozygote (AG) over homozygote allele (AA) combination in the tested clones. Gene annotation of the monoploid sugarcane genome R570 suggested that six putative candidate genes were co‐located with significant markers associated with brown rust resistance in sugarcane. The putative candidate genes regulated the formation of a cell wall barrier that plays a crucial role in controlling brown rust pathogen infection. The results of this study will open the path to exploiting new resistance sources for brown rust resistance in commercial sugarcane.

Effective Identification and Highly Sensitive Quantification of Fructo-oligosaccharide Isomers with Bi2Se3 Nanosheet-Assisted Laser Desorption Ionization Mass Spectrometry.

The growing interest in fructo-oligosaccharides (FOSs) necessitates the effective monitoring of product quality. Identifying and quantifying FOS isomers from the same sources are challenging. Here, we report a new method using Bi2Se3 nanosheets as the matrix for matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), achieving effective differentiation of oligosaccharide isomers through MALDI-MS/MS. Notably, four isomers of pentasaccharides and two isomers of heptasaccharides were successfully identified, with a remarkably low limit of detection of 0.06 pmol. Our approach enabled the specific quantification of 1F-fructofuranosylnystose in commercial FOS products, positioning it as a promising tool for oligosaccharide isomer quantification in nutritional food products. Furthermore, this technique facilitates the rapid and sensitive detection of various saccharides and a wide range of other small molecules with enhanced signal intensities and improved reproducibility. Overall, it facilitates the rapid, selective, and sensitive detection of various saccharides and other small molecules, enhancing analytical chemistry and food science applications.

Purification of T7 RNA polymerase for large scale production of mRNA vaccines and therapeutics

The COVID-19 Pandemic has revolutionized the field of mRNA vaccine technology, highlighting its utility and rapidly expanding potential, propelling it to the forefront of future vaccines & therapeutics for cryptic diseases. T7 RNA Polymerase (T7 RNAP) is a high-fidelity enzyme used for the synthesis of single-stranded mRNA from DNA templates using In vitro transcription (IVT). T7 RNAP is now the mainstay for the development of mRNA vaccines and therapeutics, accounting for 34 % of the overall raw material cost for mRNA manufacturing. Although small-scale purification processes for T7 RNAP exist, scalable process platforms and comprehensive analytical characterization are still lacking. Moreover, to address global raw material supply chain issues and to facilitate the efficient and large-scale production of mRNA vaccines & therapeutics, here we report a three-step purification process platform, yielding ∼ 85–100 mg/L of T7 RNAP from E. coli fermentation harvest. Also, to demonstrate its high-level purity and structural integrity, we have characterized it using orthogonal analytical techniques. Furthermore, when evaluated in an IVT system, this highly pure T7 RNAP showed activity that was comparable to the commercial T7 RNAP in producing integral RNA (10 kb) from a linearized plasmid DNA template. Collectively, these findings establish a purification process platform for the commercial production of T7 RNAP enzyme to support the future development of mRNA-based vaccines and therapeutics.

Akkermansia muciniphila: biology, microbial ecology, host interactions and therapeutic potential.

Akkermansia muciniphila is a gut bacterium that colonizes the gut mucosa, has a role in maintaining gut health and shows promise for potential therapeutic applications. The discovery of A. muciniphila as an important member of our gut microbiome, occupying an extraordinary niche in the human gut, has led to new hypotheses on gut health, beneficial microorganisms and host-microbiota interactions. This microorganism has established a unique position in human microbiome research, similar to its role in the gut ecosystem. Its unique traits in using mucin sugars and mechanisms of action that can modify host health have made A. muciniphila a subject of enormous attention from multiple research fields. A. muciniphila is becoming a model organism studied for its ability to modulate human health and gut microbiome structure, leading to commercial products, a genetic model and possible probiotic formulations. This Review provides an overview of A. muciniphila and Akkermansia genus phylogeny, ecophysiology and diversity. Furthermore, the Review discusses perspectives on ecology, strategies for harnessing beneficial effects of A. muciniphila for human mucosal metabolic and gut health, and its potential as a biomarker for diagnostics and prognostics.

Antimicrobial and antioxidant activities of Bromelin Propolis Spray

The indiscriminate use of antibiotics over the last 60 years has led to the development of resistant strains, leading an urgent need for new treatments, new molecules or combinations of products, safety and effective. In this regard, propolis has been used in traditional medicine due its therapeutic effects, such as antibacterial, antifungal and antiseptic, tested in vitro, in vivo and clinically. The aim of this study is to evaluate the antimicrobial and antioxidant activities of a commercial oral flavoring product against bacterial strains of clinical interest from the upper respiratory tract. Bacteria sensitivity against Bromelin Propolis Sprayâ (BPS) was evaluated using macrodilution in broth and no turbid tubes plated on TSA to confirm the results. Tested bacteria were Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa. BPS showed antimicrobial activity for all tested strains and the MBC was 2.5mg/mL. Moreover, the antioxidant activity was determined by the ability to scavenge the stable radical DPPH. 0.007% of BPS showed the maximum antioxidant effect, of 6.63mM, which demonstrate its high antioxidant effect. Therefore, these results indicate that the commercial product BPS at 0.7% can be a potential candidate as adjuvant in antibiotic therapy for upper respiratory tract disorders, due possible antimicrobial effect and its high antioxidant potential.

Determination of nut varieties and their detection in festive cookies by MALDI-TOF mass spectrometry.

Nuts contain a large amount of essential fatty acids, amino acids, and whole range of minerals and vitamins valuable for human health, yet certain risks are associated with their consumption, of which allergic reaction is the most important. Considering the growing number of people suffering from allergies caused by allergens of protein origin, the aim of this work is to find out whether nuts can be distinguished from each other on the basis of contained proteins. A total of eleven raw and subsequently heat-treated nuts (almonds, Brazil nuts, cashews, coconuts, hazelnuts, macadamia nuts, peanuts, pecans, pine nuts, pistachios, and walnuts) were analyzed using MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry with the subsequent finding of characteristic m/z values for each analyzed nut. No previous method for protein extraction was used. The characteristic values were used to verify the composition of seven types of festive cookies - six commercial products and one "unknown" cookie, where it was not known in advance, which nut it was made from. The procedure, together with the found characteristic m/z values, could serve to rapidly identify the plant origin of nut products.

Missing the biodiversity for the bee: Natural land management strategies impact functional invertebrate diversity in commercial cranberry production

Abstract Simplification of agricultural environments is linked to declines in biodiversity. Improving the floral diversity within and around these areas may result in more robust and diverse ecosystems. We investigated how floral resource abundance, diversity, and species composition in a cranberry agricultural system correlated to the abundance and overall invertebrate diversity and to the abundance and diversity of specific invertebrate groups of agricultural importance (e.g. parasitoids, phytophagous taxa, pollinators and predators). This study focused on habitats immediately surrounding cranberry production and included grassy dikes under a managed system (‘dike’), and semi‐natural areas growing on the surrounding support land (‘semi‐natural’). Floral resource availability and diversity tended to be similar between habitats, while invertebrate richness, diversity and composition differed. As the availability of floral resources increased, invertebrate abundance increased but diversity decreased. Overall invertebrate community composition differed with the specific species and availability of floral resources. The habitat type and floral resource composition impacted some agriculturally important groups, as pollinator abundance was higher in the semi‐natural habitat, and parasitoid abundance varied with floral resource composition across both habitats. These results suggest that managing the structural and floral resource diversity associated with agroecosystems can help support local biodiversity. However, these systems may disproportionately benefit more common taxonomic groups. The difference in responses of individual taxonomic groups also highlights the potential tradeoffs of focusing on only a subset of biodiversity aspects.

Organoleptic characteristics of high‐protein snacks with novel and sustainable ingredients: Cricket flour and carob powder

AbstractTo fulfill consumer trends in sustainable and healthy food choices, this study explored the application of edible insects and carob powder as sustainable and nutritious ingredients in developing a high‐protein snack, known as a protein ball. Four formulations were developed and characterized in terms of moisture content, water activity, color, texture, microbial count, and nutritional profile. Finally, the sensory profile was determined using the flash profile method, and the developed product was compared to a commercial product. The effect of replacing the conventional protein source with cricket flour and cocoa with its sustainable alternative, carob, on the measured characteristics was determined. The results showed that cricket flour significantly decreased the lightness color values (from 40 to 30) on the internal surface of the protein ball. Texture remained largely unaffected initially; however, after 2 weeks of storage, cricket flour significantly decreased the hardness (from 15 to 12 N) and chewiness values (from 1.6 to 1.0 N mm). Moreover, cricket flour significantly increased the aerobic count (from 3–4 log to 5 log cfu g−1). The sensory space of the cricket samples was separated from the milk protein samples, mainly related to flavor attributes, while the commercial sample was distinguished by dryness and sweetness. In general, carob powder did not affect the measured parameters compared to cocoa. This study demonstrated the suitability of utilizing cricket flour and carob powder in a high‐protein snack without substantially compromising the product's organoleptic properties. Future research could investigate the consumer acceptability of the product.

New Insights for High-Throughput CO2 Hydrogenation to High-Quality Fuel.

In the case of CO2 thermal-catalytic hydrogenation, highly selective olefin generation and subsequent olefin secondary reactions to fuel hydrocarbons in an ultra-short residence time is a huge challenge, especially under industrially feasible conditions. Here, we report a pioneering synthetic process that achieves selective production of high-volume commercial gasoline with the assistance of fast response mechanism. In situ experiments and DFT calculations demonstrate that the designed NaFeGaZr presents exceptional carbiding prowess, and swiftly forms carbides even at extremely brief gas residence times, facilitating olefin production. The created successive hollow zeolite HZSM-5 further reinforces aromatization of olefin diffused from NaFeGaZr via optimized mass transfer in the hollow channel of zeolite. Benefiting from its rapid response mechanism within the multifunctional catalytic system, this catalyst effectively prevents the excessive hydrogenation of intermediates and controls the swift conversion of intermediates into aromatics, even in high-throughput settings. This enables a rapid one-step synthesis of high-quality gasoline-range hydrocarbons without any post-treatment, with high commercial product compatibility and space-time yield up to 0.9 kggasoline ⋅ kgcat -1 ⋅ h-1. These findings from the current work can provide a shed for the preparation of efficient catalysts and in-depth understanding of C1 catalysis in industrial level.

Bulbs to blooms: advancements in sustainable lilium farming

The commercial cultivation and marketing of cut flowers presents significant opportunities. The market for exotic cut flowers is expanding day to day. Lilium holds a significant position in cut flower industry and is recognized as one of the top ten major varieties in the global floriculture sector. Its cultivation in non-traditional areas needs to be explored to meet the increasing demand. Hence, different research work has been undertaken in different places in order to standardize the cultivation of Lilium in open conditions. However, its cultivation is now being done under protected structures for better quality and yield. Furthermore, the practice of protected cultivation makes it possible to grow Lilium in non-traditional areas. An effort has been made to compile the production techniques developed for the commercial production of Lilium, such as higher yielding cultivars, suitable propagation techniques, intercultural operations, insect-pest disease management and post-harvest technology. These techniques will be replicated in non-traditional areas after proper validation.

Source, Extraction, Properties, and Multifunctional Applications of Pectin: A Short Review

Pectin, a heteropolysaccharide derived from plant cell walls, is essential in the food, pharmaceutical, and environmental industries. Currently, citrus and apple peels are the primary sources for commercial pectin production. The yield and quality of pectin extracted from various plant sources significantly differ based on the extraction methods employed, which include physical, chemical, and biological processes. The complex structures of pectin, composed of polygalacturonic acid and rhamnogalacturonan, influence its physicochemical properties and, consequently, its functionality. As a common polysaccharide, pectin finds applications across multiple sectors. In the food industry, it acts as a gelling agent and a packaging material; in pharmaceuticals, it is utilized for drug delivery and wound healing. Environmentally, pectin contributes to wastewater treatment by adsorbing pollutants. Current research focuses on alternative sources, sustainable extraction methods, and multifunctional applications of pectin. Ongoing studies aim to enhance extraction technologies and broaden the applications of pectin, thereby supporting sustainable development goals.

Genetic control of Fusarium circinatum tolerance in Pinus patula × P. tecunumanii hybrid families

The pine pitch canker fungus, Fusarium circinatum, has caused large-scale mortality of Pinus patula crops in South African nurseries and high in-field post-planting mortality. Tree breeders have identified selection and breeding of tolerant material as the likely long-term solution to this disease. In this study, the greenhouse artificial inoculation screening method was employed to investigate the genetic control of F. circinatum tolerance in a factorial mating design of Pinus patula × P. tecunumanii low elevation (PPTL) and high elevation (PPTH) hybrid families and their parental species. Results indicated that F. circinatum tolerance is under moderate genetic control (h 2 between 0.13 and 0.36) as assessed in a hybrid factorial mating design with a large number of PPTL and PPTH families. The PPTL hybrid displayed the highest level of tolerance, followed by pure P. tecunumanii LE, then the PPTH hybrid, pure P. tecunumanii HE and lastly pure P. patula. A general hybridising ability (GHA) and specific hybridising ability (SHA) analysis was also conducted to rank parent families as well as hybrid families. These breeding values can be used to identify superior families for commercial production and further breeding. A poor correlation between frost tolerance and F. circinatum tolerance was also found when compared to a previous frost tolerance study utilising the same hybrid families. The results from this study indicated that it is possible for breeders to cross selected P. patula seed parents with selected P. tecunumanii pollen parents to develop F. circinatum tolerant hybrid families for commercial production.

Investigation of Coffee Instant Premium by Subcritical Water Hydrolysis: Enhance of Caffeine and Antioxidant Contents

The current study observed a basic investigation into instant coffee production through subcritical water (SBW) hydrolysis. A series of SWB research in lab-scale were carried out to optimize the product qualities by adjusting the process parameters such as temperature, solvent amounts, and time for hydrolysis. The caffeine, antioxidant, pH changes, and morphologies of modified and unmodified coffee powder (commercial products) were also evaluated by SEM. The result showed that the optimum conditions were observed for SBW process at 170°C/11.18 ml/20 min with obtained 0.48 mg/ml and 29.40 µg/ml for antioxidant and caffeine, respectively. SEM images represented physical degradation of coffee extracted powder during hydrolysis. Further, the amount of coffee in the blend with mineral amount could be important sources of minerals to the human diet, as evident by EDX analysis. Further, the change of chemistry surface was also confirmed by FTIR. In summary, flow-through SBW hydrolysis of coffee extract promises for enhance caffeine and antioxidant.

Alternative products for breaking dormancy in ‘Reubennel’ plum in subtropical regions

Abstract Plum is a temperate fruit tree and thus shows abundant vegetative bud percentage and flowering following extended periods of low winter temperatures. When adequate periods of low temperatures do not occur, it is necessary to apply products to stimulate bud dormancy break and uniform leaf and flower appearance. This application is more frequently needed when plum trees are grown in the tropics. The objective of this study was to evaluate alternative products for breaking dormancy in ‘Reubennel’ plum in subtropical regions. The experiment was conducted at a site located at 22°22’S, 43°77’W and 1,173 m altitude, with a Cwa climate type. The following treatments were applied to eight-year-old ‘Reubennel’ plum trees in the 2018 and 2019 production cycles: 1) a negative standard composed of only water (control), a positive standard composed of 2) 1.5% hydrogen cyanamide (HC) (Dormex® commercial product) plus 4.5% mineral oil, 3) Erger G® organomineral fertilizer supplemented with 3% calcium nitrate, 4) 5% potassium nitrate and 5) 0.3% copper sulfate. Two liters of the treatment solutions were applied per plant with a sprayer. The effect of the tested products on vegetative bud percentage and flowering capacity, production cycle reduction and production and quality of ‘Reubennel’ plums was evaluated. It was concluded that the chemicals used to break dormancy affected only the phenology of plum trees in this subtropical region and that the application of copper sulfate can be used to break dormancy in these trees.

Catalytic Transformation of Biomass into Sustainable Carbocycles: Recent Advances, Prospects, and Challenges.

Organic compounds bearing one or more carbocycles in their molecular structure have a discernible presence in all major classes of organic products of industrial significance. However, sourcing carbocyclic compounds from exhaustible, anthropogenic carbon (e.g., petroleum) raises serious concerns about sustainability in the chemical industries. This review discusses recent advances in the renewable synthesis of carbocyclic compounds from biomass components following catalytic pathways. The mechanistic insights, process optimizations, green metrics, and alternative synthetic strategies of carbocyclic compounds have been detailed. Moreover, the renewable syntheses of carbocycles have been assessed against their existing synthetic routes from petroleum for better perspectives on their sustainability and technological preparedness. This work will assist the researchers in acquiring updated information on the sustainable synthesis of carbocyclic compounds from various biomass components, comprehending the research gaps, and developing superior synthetic processes for their commercial production.

Impact-Resistant Adhesive Based on the Shear-Stiffening Effect via Dynamic B-O Linkages.

The occurrence of adhesive failure under impact poses significant risks, including property damage, structural collapse, and even loss of life. Herein, we have developed a series of impact-resistant adhesives by incorporating dynamic B-O linkages into linear polymeric chains. These adhesives not only possess broad-area adhesion and superior adhesive strength compared to 3 M commercial products but also exhibit a shear-stiffening effect. The shear-stiffening effect provided by the B-O linkages endows the adhesives with remarkable impact resistance, achieving a force attenuation efficiency of 84.3-86.3%. Additionally, when they are bonded to target materials, the resulting sandwich structures retain their excellent impact resistance. Therefore, this class of impact-resistant adhesives with a shear-stiffening effect shows significant potential for applications in protecting precision instruments and buildings.

Chemical Profiling, Enzyme Inhibitory Activity and Antioxidant Capacity of South African Herbal Teas: Buddleja saligna, Lippia javanica, L. scaberrima and Phyla dulcis.

Herbal teas are used in South Africa as digestives to lower glycaemia and for other indications. However, the chemical composition of the infusions and their effect on enzymes related to metabolic syndrome is poorly known. The composition of infusions and methanol extracts of B. saligna (Scrophulariaceae), Lippia javanica, L. scaberrima, and Phyla dulcis (Verbenaceae) was assessed and the effect of the infusions and extract was determined towards α-glucosidase, α-amylase, and pancreatic lipase. The commercial herbal products were extracted separately with MeOH or hot water to obtain the extract/infusion for comparative studies. Total phenolic, total flavonoid and antioxidant capacity were assessed. The fingerprints of the MeOH extracts and infusions were compared by HPLD-DAD. The extract constituents were tentatively identified by HPLC-MS/MS and NMR analyses. From the extracts/infusions, 57 compounds were identified, including iridoids, phenylpropanoid glycosides, flavonoids, and caffeic acid derivatives, among others. The MeOH extracts and infusions showed strong inhibition towards α-glucosidase with IC50 in the range of 0.13-0.84 µg/mL for the phenolic-enriched infusion extract (PEI) and 0.47-0.50 µg/mL for the MeOH extracts, respectively. The P. dulcis PEI showed higher inhibition towards α-glucosidase, and the MeOH extract of L. scaberrima was better than the PEI. At 100 µg/mL, the PEI from the four herbal teas reduces the activity of α-amylase by 23.03-28.61%, with L. javanica as the most active tea. Three of the species are high in phenylpropanoid glycosides, while P. dulcis contains rosmarinic acid. Some 26 compounds were identified in the infusion from B. saligna, 28 from L. scaberrima, and 21 from P. dulcis. Four of them are common in all the teas, namely decaffeoylverbascoside, verbascoside, isoverbascoside, and tuberonic acid hexoside. Ten compounds occur in two of the teas and seventeen, fifteen, and eleven compounds were detected only in B. saligna, L. scaberrima, and P. dulcis, respectively. Most of the compounds are reported for the first time from the crude drug infusions. The results give some support for the traditional use of herbal teas as digestives and/or indications for diabetes. The chemical fingerprints set the basis for quality control of the crude drugs, based on the main constituents and differential compounds occurring in the samples.