Recent updates and perspectives of plasma in food processing: a review

ABSTRACT The demand for minimally processed food is increasing across the globe. In this regard, the application of non-thermal technologies for food processing is getting more attention. The use of plasma in food processing offers promising potential owing to its different attributes including non-thermal, enzyme inactivation, removal of pesticides toxin, less damage to food, low nutritional loss, and high quality of the final products. Plasma ensures food safety by inactivation of potential pathogens without distressing the quality indices of the food. Plasma is an advanced technique that has attracted scientists and technologists because of its astonishing capacity and potential applications in food industries which include food decontamination, inactivation of food-borne pathogenic microbes, and food packaging. The present review summarizes the key factors in plasma generation, its characteristics and applications in different food industries. Furthermore, it also addresses the principles, practices and limitations of plasma application in food safety, decontamination and current knowledge on the subject matter. These application of plasma in food industries improves the nutritional and textural aspects of food materials


Introduction
Plasma is generally defined as an ionized gas (wholly or partially), mainly comprised of atoms, ions, free electrons, and photons in their excited or fundamental states. Plasma is composed of a net neutral charge because of an equal number of positive and negative charges on it. [1] All plasma particles are referred to as "heavy species" while protons and electrons are termed as "light species." According to the phenomena of change in energy level from its solid phase to liquid, gas and finally plasma, thus bearing exceptional properties to be stated as the forth form of matter.
Due to electron different energy states plasma are divided into two categories one is thermal plasma (TP) and second is non thermal (NTP). Non-thermal plasma is attained at reduced or atmospheric pressures (near ambient temperatures of 30-60°C) and involves low power. It doesn't possess a thermodynamic equilibrium but a significantly greater electron temperature than the macroscopic

Food decontamination
Because bacteria's lack of ability to give or create resistance, plasma's multi-species nature gives a particular advantage. [4] As a result, over the last few years, a substantial amount of research has concentrated on the decontamination of food. The food material showed changed behavior toward microorganism decontamination. Pignata et al. [5] collected the data on the efficiency of plasma for food disinfection and found that 40% of previous literature exhibited the use of cold plasma on fresh vegetables and fruits, 21% of use on seeds, dry fruits, and nuts, 19% on cold cuts and meat, 10% on spices, 6% on liquid items and 4% on eggshells. Moisture content, Surface roughness, and chemistry are those product properties revealed to be factors that influence both process performance and technology appropriateness. Plasma will very certainly prove to be suitable for certain products but not for others. Food decontamination has the most impact on the food sector, but it also has the most obstacles in terms of process efficacy and regulatory approval. The use of plasma for the modification of food qualities, in which novel and desirable functional properties are induced or improved by plasma therapy, is a burgeoning area of research. Thirumdas et al. (2017) [6] provide an overview of plasma treatment of native starch to improve its functional characteristics.

Food properties modification
Cross-linking and depolymerization of side chains of amylopectin and amylose cause the characteristics to change. It has been observed that plasma treatment reduces viscosity, molecular weight, and gelatinization temperatures. Plasma etching also boosts the starch granules' hydrophilicity and raises their surface energy. Thirumdas et al. [6] Bulk mechanical properties of the resulting doughs can be improved by treating the flour. The rheological parameters of treated wheat flour demonstrated an improvement in optimum mixing time and dough strength. [7] Microbial contamination cause severe food related problems in the world and these problems addressed by the Graphene/CuO2 nanoshuttles with controllable release of oxygen nanobubbles promoting interruption of bacterial respiration. [8,9] Plasma has been discovered to alter the secondary structure of flour proteins. The findings suggested that atmospheric plasma could be used to control the functionality of wheat flour. For partial hydrogenation of soybean oil without forming trans-fatty acids, Yepeza and Keener [10] used a cold plasma (CP) discharge within a confined hydrogen gas atmosphere. Plasma is a viable alternative to classical catalytic hydrogenation in research. Former research illustrated that the surface hydrophobicity of biscuits can be improved by the application of plasma allowing improved vegetable oil spreading. [7] The effect permits the oil to retain its functionality while using less fat or oil due to the enhanced spreadability, resulting in more healthy products. Plasma-based food functionalization may confront different regulatory and technical challenges than food disinfection. However, in the food industry, cold plasma processing can hinder sensory, regulatory, and safety evaluation. [11]

Seafood decontamination
Oysters, shrimp and salmon are among the most common contributors of foodborne illness worldwide. PAW-ice was utilized by Liao et al. (2018) to preserve fresh Metapenaeusensis. After 9 days at 5℃, the shrimps' net viable count went from 3.9 log CFU/g to 6.5 log CFU/g when treated with PAWice, while it increased to 8.6 log CFU/g in case of tap water ice-treated samples. Furthermore, PAW-ice slowed the progression of melanosis in shrimps without affecting their overall quality. PAW-ice has also traditionally been used to preserve salmon slices. After 5 days of storage of PAW-ice treated salmon strips at 4 C, L. monocytogenes showed the final level of 3.6 log CFU/mL, much less than the (control) 5.1 log CFU/mL value of sterile water-ice treated samples. It has also been shown to successfully inhibit the rise of pH and levels of volatile basic nitrogen (TVB-N) in fresh salmon strips. Zhao, Ojha, et al. (2020b) [12] illustrated that PAW treatment reduced the number of P. fluorescens by 0.4 logs incubated on fresh mackerel fillets. As a result, PAW and PAW-ice proved to be successfully utilized to preserve and decontaminate seafood (Kulawik and Tiwari, 2019). [13] PAW in processed foods PAW has shown to be affective in preservation and decontamination of ready-to-eat meals and processed goods including tofu, [14] bean curd (thin sheets), [15] salted kimchi cabbage, [16] and Korean rice cakes. [17] The microbiological profile of Chinese shredded cabbages (salted) was examined using PAW washing. [16] After subjected to PAW washing, Lactic acid bacteria had a maximum alleviation of 2.2 log CFU/g, mesophilic aerobic bacteria of 2.0 log CFU/g, yeasts and molds of 1.8 log CFU/g, coliforms of 0.9 log CFU/g and L. monocytogenes had a maximum decrease of 1.5 log CFU/ g respectively. PAW washing had no affect on the moisture content of Chinese salted cabbage, but it did reduce the color values, instrumental hardness, and sugar content. Han et al. (2020) [17] recent observations have shown that a 20-minute PAW treatment on Korean rice cakes reduced the number of Penicillium chrysogenum, Candida albicans and total aerobes by 1.00, 1.97, and 2.78 log CFU/g, respectively, which was significantly statistically superior than samples treated with distilled water. Furthermore, PAW successfully decreased the incubated foodborne pathogens count on Korean rice cakes, such as L. monocytogenes, S. Typhimurium and E. coli O 157 : H 7 . However pH, hardness and color values of Korean rice cake were not significantly affected, according to the authors. In East Asian countries, products like thin sheets of bean curd (soy-based foods) and tofu are the most popular and well-known. Tofu is a satisfactory medium for microbial growth development because of its nearneutral pH, and high water and protein content. Microbial loads on tofu samples treated with PAW were examined to be decreased during storage by 0.5-0.8 log CFU/g after 24 hours, according to. [14] PAW-treated tofu samples might keep their fresh color and texture. Similar results was shown PAW treated thin sheets of bean curd (a classic Southeast Asian bean product). [15] PAW important for microbial decontamination of food products, pesticide residue reduction, meat curing, sprout production, and food contact material disinfection. [18 Table 1 showed the PAW-based microbial decontamination of various food components whereas Table 2 showed the impact of non-thermal plasma treatment on food functional components.

Sprout production
Sprouts have traditionally been consumed for centuries as nutritionally significant vegetable. PAW has a lot of potential for sprout production because of its strong antimicrobial properties and a greater number of reactive species. PAW was found to efficiently boost the rate of germination of soybean seeds and greatly improve the level of c-aminobutyric acid, ascorbate and asparagine in sprouts. These results are in accordance with references. [19,20] A previous study evaluated that seed sprouting requires humid and warm conditions, which are perfect for improved growth and better survival of foodborne pathogens such as S. aureus, E. coli O 157 :H 7 , B. cereus, L. monocytogenes and S. Enteritidis. Recent studies showed that PAW watering has also been shown to efficiently reduce microbial burdens on sprouts. Microbial counts dropped by 5.17, 4.29, 2.80, and 2.04 log for sprouts treated with Air, O 2 , He, and N 2 -PAW, respectively. [19] As a result, as previously stated, PAW could contribute greatly to the cost-effectiveness and hygiene of sprouts production.

Potential food industrial applications of plasma and PAW
In recent years the application of PAW has been extensively explored in the food sector including meat curing, fruit and vegetable preservation, eggshell and meat decontamination, and pesticide reduction. There are many potential applications for plasma in the food industry, each with its own set of benefits and obstacles. Food contact surfaces and packaging materials have relatively homogeneous and smooth surfaces, allowing for easy treatment and a well-defined regulatory acceptance pathway.
Over the past few years, the use of plasma technology has become significant worldwide. Plasma's low-temperature characteristics and high effectiveness in the deactivation of microbes make it excellent for usage in food sector applications. Concerning foods and food-related materials, plasma treatment proposes many options in food processing, such as improvement of mass transfer, decontamination of surface areas, and alteration of surface properties. This technique has also been used to effectively sterilize packaging materials as well as modify their functionality to obtain desired characteristics. [1,7]

Cereal industry
More than half of the supplies and crops included in cereals are used for consumption. A potential non-thermal method used in the cereal industry is plasma technology for a variety of crops. Besides decontamination, this method has many advantages in the cereal business. Brown rice is not as popular as white rice because of its poor eating properties and cooking characteristics. Brown rice, on the other hand, is favored due to its higher nutritional worth. Brown rice undergoes plasma processing to change its characteristics. A past researcher modified the textural aspects, cooking characteristics, and microstructure of brown rice using low-pressure plasma. Treatment of plasma causes an etching surface of the brown rice, facilitating faster water absorption while soaking rice. Brown rice takes less time to cook after treating, and the cooked brown rice is easier to chew and has a softer texture. Brown rice's nutritional value was also improved using the plasma process. [21] In comparison to control treatments, the plasma process was observed to enhance the antioxidant potential in brown rice. Figure 1 shows the potential used of plasma in food application. Wheat seed growth has been demonstrated to be aided by plasma treatment. Atmospheric cold plasma processing on soft and hard wheat flour was examined by Misra et al. (2015). [7] The rheological characteristics of flours demonstrated an enhancement in optimal mixing time and strength of the dough for both weak as well as strong wheat flours. Corn starch granule morphology, rheological characteristics, and molecular and crystalline structure are all affected by dielectric barrier discharge (DBD) plasma. Besides, modifying starch granules' surface area, DBD plasma also entered their core, thus resulting in the molecular breakdown, oxidation of partial hydroxyl to carboxyl groups, wider channels, and a loss in crystallinity due to pinhole structures present in the starch granules. [22] The findings revealed that the food quality of beans and wheat was unaffected or just slightly changed after plasma treatment. After plasma processing, the seeds were found to be viable properly.
Treatment efficiency improves as treatment time, frequency and voltage are increased. Plasma is an innovative and capable insect control strategy to be used for stored crops (cereals). With decreasing distance from the nozzle point and increasing plasma jet pulses, a significant decrease in adult emergence and increases in pupal and larval mortality were observed. Larvae were more responsive to the process treatment than pupae, but the point of concern is that the treated pupae produced more deformed adults than treated larvae. According to the research, cold plasma induces oxidative damage in P. interpunctella larvae, this is because of the stress caused by reactive oxygen in their bodies . [23] In comparison to control larvae, treated larvae had significantly lower levels of protein and glutathione content as well as higher levels of lipid peroxides. Raw milk obtained from the dairy industry is found to be a natural and nutrient-enrich product that can be used as a significant and quick nutritional supplement for human consumption. Appropriately treating raw milk for eliminating harmful pathogens before ingestion is therefore a standard protocol to be used either by using UHT treatment or pasteurization process. Current thermally treated decontamination treatment has been shown to alter the physiological and chemical characteristics of milk and various milk products. Milk has proven to be particularly sensitive to several innovative technologies due to its complex structural behavior. [24]

Meat processing industry
The meat and poultry sector has been at the forefront of developing and executing innovative processing methods. Fabrication of meat products (i.e., cutting, extruding, and blending), Slaughtering, processing of meat products (i.e., drying, curing, cooking, and freezing), and packaging are among the food safety intervention techniques used in this industry. Intervention techniques are also used to certify a sanitary manufacturing environment. Plasma technology provides a diverse range of operating methods, product exposure methodology, and composition of the gas that ultimately affect the resultant reactant product's concentration (i.e., monoxides, ozone, and peroxides) and ultimately to the meat and poultry sector. Plasma treatment can preserve swine musculus longissimus dorsi in terms of microbial contamination for a long period and also showed its capability to decontaminate fresh pork. The utilization of DBD plasma technology was investigated by Kim et al. (2013) [25] to enhance the shelf life of pork loins. E. coli population was decreased by log cycles of 0.26 and 0.50 after treating for 5 minutes and enhanced reduction was observed after 10 minutes of treatments i.e., log units 0.34 and 0.55 when DBD plasma applied or exposed to pork loin along with the input gases such as He and He + O2 respectively.
When the specimens were underwent DBD for 5 and 10 minutes with He and He + O2, a decrease in the concentration of L. monocytogenes was observed from 0.17 to 0.35 and 0.43 to 0.59 log cycles respectively. Using DBD plasma processing, potential reduction in Lightness (L*) values and pH of the sample was observed but at the same time no significant change in redness (a*) and yellowness (b*) was investigated. In comparison to other samples, lipid oxidation was higher in samples with He + O2. Sensory attributes such as color, acceptability, appearance, and odor are evaluated to decrease in samples treated with DBD. [25] These research results have shown that the DBD plasma technique could be used to decontaminate pig loins by deactivating pathogenic bacteria.

Fruit and vegetable industry
Plasma is a significant technique to be used in the processing industry of fruit and vegetable. A microbial count can be reduced on these surfaces by plasma technology. One of the significant applications of plasma is the decontamination of freshly produced food products stored in a sealed pack. Plasma can be directly produced inductively in the container utilizing the DBD along with suitably ordered electrodes. The impact of DBD on strawberries was investigated previously. Strawberry background microflora comprised fungi (yeast, molds) and mesophilic aerobic bacteria that were decreased by 2 log cfu/g within 5 minutes. The plasma treatment showed no noticeable effect on color and firmness of strawberries. [26] Between treated and control samples. No significant changes in antioxidants (i.e., polyphenol and ascorbic acid) and texture were investigated. After plasma processing, fresh dragon fruit with green tea showed greater levels of crude fiber, crude protein, total phenolic content (TPC), and crude fat. [27] The findings suggested that combining atmospheric plasma and extracts of green tea could prevent fresh dragon fruit from pathogen growth and increase its shelf life without compromising sensory attributes and nutritional quality. Research conducted by Lacombe et al. [28] observed the deactivation of aerobic microbes on blueberries by using the effect of atmospheric non-thermal plasma on quality characteristics. This processing technique proved to be an efficient technology to decontaminate leafy vegetables. Non-thermal oxygen plasma processing was observed to possess a great impact on the effective sanitization of surfaces of fresh produce. Depending on the properties of different food surfaces, deactivation of S. typhimurium on different fresh foods using plasma processing was observed. Nutritional properties could also be enhanced using plasma treatment. Figure 2 showed the effect of plasma on food quality.
Pasteurized pomegranate juice was compared to the juice treated with gas-phase plasma to investigate the contents of phenolic compounds. [29] Plasma and pasteurization treatments showed a significant increase of 33.3 and 29.55% of phenolic compounds respectively. In comparison to pasteurized juice, plasma-treated pomegranate juice proved to hold the maximum contents of phenolic acids. Cold plasma was used to decontaminate cherry tomatoes along with maintaining the quality of the end product. Weight loss and the rate of respiration were continuously observed, while the rest of the parameters were evaluated at the last step of the storage. The application of non-thermal plasma on cherry tomatoes did not show any increase in rates of respiration. The effect of non-thermal plasma on enzyme activity has previously been studied, this technique is a remarkable technology capable of lowering the activity of dietary enzymes such as peroxidase and polyphenol oxidase. [30]

Spice industry
Plasma technology has also been used to decontaminate spices, with encouraging results. Hertwig et al. (2015a) [31] studied the influence of plasma processing on the natural microbiological count and qualitative attributes of specific spices and herbs (crushed oregano, paprika powder and pepper seeds) when exposed to plasma-treated air for up to 90 minutes, as well as the inactivation of their indigenous microbial flora, was also observed. Paprika powder's and pepper seed's native microbial flora was decreased by 60 minutes of exposure to remote plasma processing for greater than 3 logs CFU/g. Owing to the breakdown of carotenoids, After ≥5 minutes of remote plasma treatment on paprika powder results in a significant decrease of redness. Another study looked into the antibacterial effects of two different atmospheric pressure plasmas on the decontamination of whole black pepper. A microwave-driven remote plasma or plasma jet was used to treat peppercorns (naturally contaminated) inoculated with spores of B. atrophaeus, B. subtilis and S. enteric. Direct cold atmospheric pressure plasma processing exhibited a reduced rate of deactivation, this could be the reason for multiple deactivation processes and peppercorn's complex surface structure. Remote plasma resulted in 2.4, 3.8, and 4.1 logs CFU/g decreases of spores of B. subtilis, B. atrophaeus and S. enteric respectively, (after half-hour of processing). [32] But similar deactivation results didn't obtain from the direct application of the plasma jet. It was also evaluated that quality characteristics including contents of volatile oil, piperine, and color properties were not affected primarily.

Effect of plasma on food quality
The success of CP treatment on food stuffs is also measured by how CP affects the product's aroma, flavor, and color. Depending on the exposure parameters, time duration, and CP process residues, specific parameters i.e., quality characteristics including the sensory and nutritional properties could alter. [33] Because of its reduced penetration in the food, only the food surfaces are affected which helps to maintain the nutritional profile of the product. Generally, the CP technique is not considered to hinder the nutritional profile of food products. Based on the type of the product, it is important to note that this effect is extremely variable and thus optimizing of product is required for various food products. CP has influenced various quality parameters as discussed below.

Effect on organoleptic properties (color, flavor, and texture)
Previous literature showed that when CP processing was applied to pepper, it was observed that the color properties did not alter. Despite this, some research studies also evaluated that some alterations in the color of the product showed significant appearance properties. There was an observed increase in the redness of red chicory by [28] evaluated the darkening in color of blueberries. A decrease in a* values and an increase in L* and b* values of milk were observed by. [34] Color changes in tomatoes and carrots, as well as photosynthetic activity in fresh maize and cucumber, were noted by. [35] Blueberries' firmness (texture) was also found to reduce by CP treatment, [28] but the firmness of cherry tomatoes remained unchanged by CP. These negative effects on quality attributes boosted the demand for CP parameterization for a specific product type.

Effect on rheological properties
Rheological properties of starch can be altered by the latest physical technique of plasma processing, specifically with high concentrated and low viscosity of green production of starch. DBD enormously influence the rheological characteristics of crystalline structure, granule morphology and molecular structure of corn starch. Pinole structure of starch granule is the reason that DBD plasma willing to alter the starch granules' surface, produce large channels as it inter to the interior and moreover, reduce the range of molecular degradation and crystallinity oxidation (hydroxyl to carboxyl groups). Although, degradation of starch cause a rise in starch molecules concentration, from non-Newtonian liquid to Newtonian fluid the properties of starch paste changed and also reduce the viscosity. [36]

Effect on bioactive compounds
Thermal processing is frequently damaging to the bioactive components of food. As a response, every food processing method must ensure that bioactive components are preserved (glutathione, antioxidants, ascorbic acid, vitamin C, -tocopherol, -carotene, and other flavonoids). Overall, CP revealed a high ability to preserve phenolic compounds and dietary antioxidant activity. Literature found that employing CP instead of traditional pasteurization increased phenolic acid and anthocyanin retention properties in sour cherry marasca juice. Hereg and co-workwes, [29] found that the total phenolic content of pomegranate juice increased by 33.03% after CP treatment, while [33] found that chicory antioxidant capacity remained unaltered. CP treatment has a minor negative influence on certain commodities while having a positive impact on phenolic compounds. [28] Observed a reduction in anthocyanin (as cyanidin3-glucoside equivalent) in blueberries treated with CP.

Effect on enzymes
The level of enzymatic inactivation caused by CP is affected by a variety of factors such as power input, types of reactive species, type of gas, nature of exposure, enzyme structure, and so on. The enzyme's secondary structure is lost as a result of CP processing due to factors i.e., side-chain alterations and bond cleavage properties by the chemically active species in cold plasma. [37] Using atmospheric air DBD plasma, a former researcher discovered a decrease in tomato peroxidase (POD) enzyme activity at various voltages. In 300 seconds at 2 kV, CP treatment of guava pulp and entire fruit reduced polyphenol oxidase (PPO) enzyme activity by 70% and 10%, respectively . [3] Although the majority of studies demonstrated that CP treatment had a favorable effect on enzyme inactivation, numerous findings also reveal that enzymatic activity causes slight or no changes after CP processing. As a result, additional research is required to thoroughly examine the interactions of substances in plasma with enzymes involved in diverse food preparation procedures.

Effect on toxins and allergens
Food-related toxicity and allergies have increased dramatically in recent decades. As a result, maintaining toxin-and allergen-free food has become a demanding task for the food processing industry. Food allergens are proteins, and studies have shown that plasma can remove proteins that are tightly bound to food matrices. [38] Thirumdas et al. [3] found that plasma treatment may have some influence on protein allergenicity in the same manner as gamma irradiation eliminated or reduced allergenicity of lectins in legumes). It was shown that peanuts and walnuts treated with plasma had an 18% increase in peroxide value because of radicals that can oxidize lipid molecules, resulting in an increase in peroxide value.

Action of plasma on microorganisms
Plasma affects microbial cell components and functions. The application of plasma's sterilizing properties was originally proposed in the late 1960s, was patented in 1968, and the first experiments using plasma produced from oxygen were proposed in 1989. Following that, Much study has been conducted on the mechanism of inactivation of microorganisms by plasma chemicals. By interacting with biological substances, plasma agents contribute to the noxious effect. Microbes in plasma are likely subjected to a barrage of radicals, resulting in surface damage that the living cell is unable to repair quickly enough. [39] This could account for some of the facts, such as how cells are frequently killed. "Etching" is the name given to this procedure, according to past literature.

Eggshell
Salmonella spp., a foodborne-pathogenic bacteria, is the most common eggshell-associated pathogen, inflicting widespread public health and economic implications. Additionally, the PAW-treated eggs exhibited better freshness indices than the chlorine-treated eggs. This study suggests that PAW may be a more efficient and effective method than commercial eggshell cleansing and disinfection techniques. [40] Bacterial contamination cause severe food related problems in the world and these problems addressed by the nanomaterials e.g grapheme/CuO 2 . The rGO/CuO2 nanocomposite was reported as an oxygen nanoshuttle capable of transferring O2 NBs in a controlled manner to combat bacterial infections. Super critical water are used to treat the bacterial Staphylococcus aureus infections. [41] Thawing media for enhancing microbiological safety One of the most common methods for preservation meat, poultry, fish, and shellfish is freezing. Before cooking or further processing, the above-mentioned foodstuffs must be thawed technically. PAW was recently examined in the frozen beef thawing by previous literature. The scientists discovered that PAW thawed beef had much lower levels of total aerobic bacteria, yeast and mold, than water, air, microwave, or minute acidic electrolyzed water thawed samples. [18] However, when compared to other standard thawing procedures, PAW-supplemented thawing had no negative impacts on beef pH, texture or color. As a result, PAW can be administered as an efficient thawing media to keep frozen products' microbiological safety and quality intact.

Plasma and food packaging
The use of plasma for packaging was discussed by Jeon and coworkers [42] and the reported it does not damages the packaging material structure as it is operated at low temperatures. It is efficient as it inhibits the chances of cross contamination that usually occurs between the application of microbial limiting treatments and the packaging process of the food product. [42] It is possible to prepare a variety of materials and coatings, some of which are very thin, using atmospheric pressure, plasma pretreatment, when creating composite packaging. [43] Testing on the impacts of CPs on food packaging materials, such as PE and PET, must be done in order to guarantee that they are within controlled limits and to determine the permeability related with O 2 and water vapor. [44]

Conclusion
The application of eco-innovative technologies has introduced emerging plasma technology used in food industries. Owing to its unique non-thermal property plasma application has attracted its application for food processing and food safety for heat-sensitive commodities. In addition to this advantageous effect on food, quality has also been observed. Plasma-activated water generation and synergistic effects of plasma with other traditional technologies have displayed promising outcomes. Thus, from the lab to the food industry translation of plasma techniques portray huge challenges as well as opportunities.

Funding
The authors have no funding to report.

Ethical approval
This study work does not contain human or animal based trial study.