Influence Of Heating Process Research Articles

Influence of heat processing on the anti-inflammatory activity of fresh Smilax glabra based on PDE4 inhibition.

Smilax glabra Roxb. (SG) is widely used as functional food with various beneficial effects. Fresh SG without processing has been eaten directly for anti-inflammation from ancient China, while the underlying mechanism remains underexplored. This study aims to investigate the anti-inflammatory activity of fresh SG by using metabolites profiles, affinity ultrafiltration mass spectrometry, PDE4 enzyme inhibition assay, and in silico analysis. Encouragingly, fresh SG showed promising anti-inflammatory effect with IC50 value (0.009μg/μL) on PDE4 was about 12 times higher than that of processed SG (0.110μg/μL). Astilbin was identified as the main bioactive compound of fresh SG responsible for PDE4 inhibitory activity. We found that heat processing strongly affected astilbin isomerization, leading to significant changes in contents and PDE4 inhibitory activities of four astilbin isomers, resulting in decreased anti-inflammatory activity of fresh SG. This finding will provide theoretical basis for systematic research and food/nutraceutical applications of fresh Smilax glabra in the future.

Metabolite identification in fresh wheat grains of different colors and the influence of heat processing on metabolites via targeted and non-targeted metabolomics

Phenolic antioxidants are phytochemical components in wheat grains that provide a variety of potential health benefits. The metabolites and antioxidant activity of fresh, mature, and heat-treated, wheat grains with black, blue, purple, and white grain coats were identified by targeted and non-targeted metabolomics. The total phenolic (TPC) and flavonoid contents (TFC) and antioxidant activity (AOA) increased with the darkening of grain color, the general trend being black > purple > blue > white. Purple and black wheat are rich in rutin (3916 µg/kg and 3066 µg/kg, respectively) and peonidin-3-O-glucoside chloride (2595 µg/kg and 1740 µg/kg, respectively), while blue wheat is rich in luteolin (2076 µg/kg). In most cases, TPC, TFC, and AOA had the greatest values in fresh grains and the lowest values in mature grains. Using non-targeted metabolomics, a total of 866 metabolites were identified in the tested fresh wheat grains, 106 flavonoids and 39 phenolic acids. In total, the relative abundance of flavonoids in purple and black wheat was higher than in blue wheat, indicating a higher nutritional value of fresh black and purple grains. After heat processing, the content of most metabolites decreased in heat-treated purple grain, whereas heat treatment significantly increased the content of peonidin-3-O-glucoside chloride (2.27-fold) and cynaroside (12.01-fold). This study clarifies that seed coat color and processing treatments impact the metabolite contents and antioxidant activity of wheat grains, providing valuable information for improving the nutritional quality of food during processing.

Effect of different drying treatments on total polyphenolics content and in-vitro biological properties of Ficus benghalensis fruit: A comparative study

The effect of different drying process on Ficus benghalensis fruits was studied for their polyphenolics, polysaccharides , antioxidant, anti-inflammatory, and antibacterial activities in-vitro. Crude polyphenol isolates were quantitatively analyzed for determination of total phenol, flavonoids , and tannin content. Antioxidant activity of crude polyphenolics was analyzed by in-vitro DPPH method. Protein denaturation assay was used to determine the anti-inflammatory activity of crude polyphenolics from F. benghalensis fruits. The antibacterial activity was determined by growth inhibition assay against Salmonella typhi . Quantitative analysis showed that micro-oven-dried fruit has high total phenolic (92.01 ± 03.46 mg GAE/g extract) and tannin contents (38.27 ± 06.03 mg GAE/g extract). Higher radical scavenging activity was registered for crude polyphenolics from shade- and hot-air-dried fruit (IC 50 at 158.75 and 165.85 μg/ml). Phenolics concentration of micro-oven-dried fruits exhibits higher level protection against protein denaturation (44.79 ± 3.26% at 0.5 mg sample) and showed lowest growth inhibition against S.typhi (1.12± 0.94% at 0.5 mg). The influence of heat processing on ficus fruit and its polyphenolics were analyzed by Fourier transformed infrared spectroscopy . Thus, shade and hot-air drying can be possible methods for obtaining heat-resistant polyphenolics with good biological activity from ficus fruits. • The polyphenolic contents of fruits were influenced by drying methods. • The shade- and hot-air-dried fruits showed higher activity. • Drying methods regulate the functional properties of F. benghalensis fruits.

Study on hydrogen behaviors around micropores within heavy forging during heating process

Hydrogen and micropores are widely distributed and inevitable in heavy forgings. The accumulation of hydrogen in micropores results in the formation of higher hydrogen pressure inside. In this article, the influence of heating process on hydrogen behavior around micropores is studied by finite element method. The analysis model is established theoretically and the relationship among micropore hydrogen pressure, temperature, and lattice hydrogen concentration is derived based on chemical potential balance. The results show that, during the heating process, when decomposition condition is met, hydrogen molecules begin to decompose and diffuse out of micropores. Micropore hydrogen pressure is the result of volume expansion and decomposition of micropore hydrogen molecules. Microstructures with a smaller hydrogen diffusion coefficient are more likely to form higher hydrogen pressure in micropores during heating and are more likely to form hydrogen-induced cracks. The holding temperature has little effect on micropore hydrogen pressure. Among all heat treatment parameters, the heating rate has the most significant influence on hydrogen behavior around micropores. A larger heating rate can reduce hydrogen discharge time, but increase the micropore hydrogen pressure. From the perspective of reducing micropore hydrogen pressure and heat treatment time, a heating rate of 0.05 K/s is more appropriate. This study puts forward a new mechanism of hydrogen-induced cracking in heavy forgings and provides a new perspective for formulating heat treatment processes.

Oil characteristics and influence of heat processing on fatty acid profile of wild harvested termite (Macrotermes subhylanus) and long-horned grasshopper (Ruspolia differens)

The aim of this study was to assess the physico-chemical characteristics and fatty acid profile of oil from termite (Macrotermes subhylanus), green and brown long-horned grasshopper (Ruspolia differens). The insects were harvested in Siaya county of Western Kenya from the wild during swarming seasons. Physico-chemical characteristics of insects oil and the effect of two traditional heat processing methods (frying and solar drying) on the fatty acid profile were determined using standard methods. The insects oil was a clear, golden or light yellow colored liquid at room temperature, with a low solidification temperature (8–12 °C) for M. subhylanus and10–15 °C for R. differens. M. subhylanus showed an iodine value of 83.51 g iodine/100 g oil, peroxide value 0.19 mEq/Kg, saponification value 160.61 mg KOH/Kg and 38.77 mg/100 ml of total cholesterol. R. differens showed an iodine value of 86.97 g iodine/100 g oil, peroxide value 0.13–0.14 mEq/Kg, saponification value 229.67–234.40 mg KOH/Kg and 27.51–31.40 mg/100 ml of total cholesterol. Oleic acid was the major fatty acid in all the fresh insect samples (M. subylanus 52.1%, green R. differens 43.2%, brown R. differens 37.1%) while palmitic acid was the second most abundant fatty acid. Total unsaturated fatty acids were proportionately more than the total saturated fatty acids. The level of saturated fatty acids increased from fresh insect samples to heat processed samples (fried, fresh-dried and fried-dried), while the level of unsaturated fatty acids decreased with processing though insignificantly. The insects have high quality oil that maybe exploited in processing of nutritional food products and they may therefore be heat processed domestically without adversely affecting the quality of the oils.

Influence of Heat Processing on DNA Degradation and PCR-Based Detection of Wild-Type and Transgenic Maize

Reliable detection of genetically modified (GM) maize is significant for food authenticity, labelling, quality, and safety assessment. This study aims to evaluate the factors influencing degradation and polymerase chain reaction (PCR) amplification of DNA from the wild type and transgenic maize (events Bt-176 and MON810) during thermal treatment at 100°C and 121°C. A new PCR method was developed targeting the Cry1Ab gene to detect insect-resistant GM plants. The yield of genomic DNAs extracted by the DNeasy plant mini kit dramatically decreased while DNAs obtained by cetyltrimethyl ammonium bromide- (CTAB-) based method did not show any visible changes in the yield by the time of processing. Treatment at 100°C did not significantly affect either genomic DNAs or amplicons. Heating at 121°C induced time-dependent degradation of genomic DNAs and exogenous Cry1Ab gene; however, it did not have any considerable influence on the exogenous 141 bp amplicons or endogenous amplicons in the range of 102 bp to 226 bp with the exception of the event MON810 extracted by the DNeasy plant mini kit. More yield was observed at 226 bp than 140 bp fragment of the invertase gene. The 141 bp fragment of the transgenic CaMV 35S promoter exhibited the highest thermal stability of all the examined amplicons. Analysis of foodstuffs demonstrated 102 bp amplicons specific for the zein gene as the effective marker to detect maize in the processed foods. The obtained results demonstrate that PCR-based detection of the wild type and transgenic maize is dependent on the combination of different parameters of crucial factors such as temperature and duration of exposure, transgenic event, DNA extraction method, DNA marker, and size and location of amplicons.

Вибір рецептурних компонентів для виробництва плодових соусів на основі ягід кизилу�

Вибір рецептурних компонентів для виробництва плодових соусів на основі ягід кизилу�

Influence of heat processing on the volatile organic compounds and microbial diversity of salted and vacuum-packaged silver carp (Hypophthalmichthys molitrix) fillets during storage

Ready-to-eat products have become popular with most of the busy people in modern cities. Heat processing combined with vacuum-packaging is one of the most common methods to make ready-to-eat products with an extended shelf-life. In this study, the influence of heat processing [80 °C (LT) and 98 °C (HT) in water bath] on the quality of salted and vacuum-packaged silver carp (Hypophthalmichthys molitrix) fillets, stored at 20 ± 1 °C, was investigated by sensory analysis, biochemical analysis, and microbial diversity. SPME-GC/MS indicated the presence of 27 volatile organic compounds (VOCs) in fillets, and major VOCs were aldehydes and alcohols. Acids tended to increase during storage and caused a fetid odor at the end of storage. Culture-dependent method indicated that Bacillus dominated the spoiled LT and HT samples. In addition, Bacillus was identified as the main spoiler of deteriorated heated fillets by high-throughput sequencing. Sphingomonas and Brevibacillus dominated the indigenous bacteria of fresh raw fillets. After heat processing, LT samples exhibited higher organoleptic quality than HT samples on day 0. HT samples showed extended shelf-life at 20 °C storage compared to LT samples.

Bioaccessibility of selenium, selenomethionine and selenocysteine from foods and influence of heat processing on the same

Selenium (Se) is an essential nutrient with diverse physiological functions. The selenium content of commonly consumed cereals, pulses and green leafy vegetables (GLV) was determined. Bioaccessibility of Se, and its organic forms selenomethionine (SeMet), and selenocysteine (SeCys2) was also examined, and the effect of heat processing on the same was studied. The bioaccessibility of Se in cereals ranged from 10% to 24%, that of pulses was between 12% and 29%, and of GLV, 10–31%. The concentration of SeMet in the dialysates of the cereals, pulses and GLV ranged from 5.15 to 28.7, 2.7 to 36.2, and 0.03 to 5ngg−1, respectively. The concentration of SeCys2 in the dialysates of the foods examined was negligible. Heat processing significantly decreased the bioaccessibility of Se, SeMet and SeCys2. This is the first report on the bioaccessibility of Se and its major organic forms from commonly consumed staples, and the effect of heat processing on the same.

The Temperature Effect on Properties of Fe-Co-Ni Nanostructured System

<p>The influence of heating process (up to 200, 400 and 600 °C) on the phase transformation of non-equilibrium biphase Fe-Co-Ni systems was studied. The system was not changed at 200 °C, it remained nanostructured, and there were no phase transformations. At higher temperatures phase transformation BCC- into FCC-phase (high-temperature phase modification) was observed. The degree of phase transformation depended on the system composition. At 400–600 °C there was coarsening of the crystallites. In addition, there was the phenomenon of system oxidizing attended by iron outflow from phase; as a result, the lattice parameters were decreased. According to the measurements of the lattice parameters at coolingdown, parameters dependences on temperature and coefficients of thermal expansion were obtained. Changing of the system composition (due to metal oxidation) had low impact on the coefficients. The phases with different compositions and close lattice parameters had different coefficients of expansion. It was supposed that the expansion coefficients dependence from the composition was not additive. Magnetic characteristics of the samples were measured. It was found that heating changed it essentially. The main reasons of it was coarsening of grains (crystallites) and changes of the phase composition. The dependence of the magnetic properties of systems on phase composition and size of the crystallites (particles), as well as changes occurring under the temperature effect were required a more detailed study.</p>

High Temperature Properties of CoFe/CoNi and Fe/CoNi Biphase Microwires

The objective of this work has been to analyze the high-temperature behavior of magnetically single-and biphase microwires because of its interest from fundamental and applications viewpoints. Two alloy compositions with amorphous structure covered by glass have been prepared as magnetically single phase microwires by quenching & drawing technique: CoFe-based with near zero saturation magnetostriction constant and Fe-based with positive saturation magnetostriction constant. The same wires were used as the core for magnetically biphase microwires. Second CoNi phase was deposited by electroplating. Magnitudes as saturation magnetization and hysteresis parameters are determined in the temperature range from room temperature up to 1200 K. We proceed to a comparative analysis of their magnetic behaviour at different temperatures as well as after cooling down to room temperature. Information on the Curie temperature of different phases and on the influence of heating process on the magnetic properties is thus derived.

Storage-induced changes in the sensory characteristics and volatiles of conventional and lactose-hydrolyzed UHT processed milk

Storage-induced changes are known to be more prominent in lactose-hydrolyzed (LH) milk compared to conventional milk. Therefore the present study aimed at identifying off-flavors resembling from formation of volatiles during storage of ultra-high temperature treated (UHT) LH milk and conventional UHT milk. Further, the influence of heat processing, indirect or direct, on UHT LH milk was also examined. Storage-induced changes in sensory attributes, volatiles and primary amines were investigated during a 4 months period. Conventional UHT milk (with 5 % lactose) processed using indirect heat treatment (CONVI) and two types of UHT LH milk (with less than 0.01 % lactose) produced using either direct heat treatment (LHD) or indirect heat treatment (LHI) were represented in the study. Sensory descriptive analysis showed that fresh samples of CONVI, LHI and LHD differed in sensory properties and the samples could be differentiated according to boiled and stale aroma as well as color saturation. Differentiation of the fresh samples based on the volatile gas chromatography–mass spectrometry profile was not achievable. During storage, samples developed differently with respect to sensory characteristics, volatiles and the amount of primary amines. Partial least squares models (PLS1) including only methyl ketones and aldehydes showed that 2-butanone, 2-pentanone, 2-heptanone, 2-nonanone, heptanal, octanal and nonanal predicted stale flavor. Bitter taste, on the other hand, correlated with the amount of primary amines (Pearson’s correlation, r 2 = 0.71). This finding indicates that storage-induced changes in sensory characteristics, volatiles and primary amines depend on both differences in lactose content and the applied heat processing.

Influence of Heat Processing on Nutrient Composition and Energy Values of Selected Cereals Consumed in Nigeria

Aims: The influence of heat processing on the nutrient composition and energy values of selected cereals (Sorghum, Maize, Millet, and Rice) consumed in Nigeria were investigated. Study Design : Cereals samples (100 g each) were weighed out for processing prior to analysis. The weighed samples were divided into two lots of raw and cooked sample. Raw sample was sun dried, further oven dried for 24 hours and ground into flour which passed through a 30 - mesh test

Organic vs. traditional potato powder

Abstract In this work the profile of primary metabolites in potato (Solanum tuberosum L., cultivar Agria) grown under organic vs traditional farming system was studied, and the influence of heat processing (for producing potato powder) on nutritionally important compounds such as essential amino acids, proteins and carbohydrates was evaluated. The potato powder was obtained by means of a drum dryer using different process parameters (temperature, time and product thickness) to minimise thermal damage on the final product. The effects of these different processes were evaluated by analysing also some other quality indicators, as final moisture, colour of the powder, and rheological properties of the rehydrated puree. Moreover, because of its potential cancerogenity and genotoxicity, acrylamide content was determined in the potato powder obtained and compared with that of its main precursors (asparagine and reducing sugars). The steam pressure of 0.2 MPa and 2 r.p.m. allowed to obtain potato powder with a lower acrylamide contents and still a very low water content that caused no microbiological safety problems. This powder, both from conventional and organic farming, when rehydrated, resulted in purees with minimal nutritional losses and colour and consistency very similar to the fresh ones.

Thermal conductivity measurements of porous dust aggregates: I. Technique, model and first results

We present a non-invasive technique for measuring the thermal conductivity of fragile and sensitive materials. In the context of planet-formation research, the investigation of the thermal conductivity of porous dust aggregates provide important knowledge about the influence of heating processes, like internal heating by radioactive decay of short-lived nuclei, e.g. 26Al, on the evolution and growth of planetesimals. The determination of the thermal conductivity was performed by a combination of laboratory experiments and numerical simulations. An IR camera measured the temperature distribution of the sample surface heated by a well-characterized laser beam. The thermal conductivity as free parameter in the model calculations, exactly emulating the experiment, was varied until the experimental and numerical temperature distributions showed best agreement. Thus, we determined for three types of porous dust samples, consisting of spherical, 1.5 μm-sized SiO 2 particles, with volume filling factors in the range of 15–54%, the thermal conductivity to be 0.002–0.02 W m −1 K −1, respectively. From our results, we can conclude that the thermal conductivity mainly depends on the volume filling factor. Further investigations, which are planned for different materials and varied contact area sizes (produced by sintering), will prove the appropriate dependencies in more detail.

Influence of heat processing on functional properties of Australian wattle seed ( Acacia victoriae Bentham) extracts

Whole wattle ( Acacia victoriae Bentham) seed was extracted with water before or after a soaking/heat treatment regime designed to destroy its protease inhibitors. The yield, composition and physical properties of these extracts were measured and they were then subjected to an analysis of their functional properties, which included emulsion and foam formation and stabilization, solubility at different pH values and gelling ability. Processing of soaked wattle seed at 100 °C for 30 s led to a much reduced extract yield and viscosity at both pH 4 and 7, decreased its water-soluble carbohydrate content but increased its protein content, and solubility under alkaline conditions. Processing of wattle seed before extraction also led to increased emulsion droplet size and reduced emulsion stability, the differences being more pronounced in emulsions formed with 20% oil compared with 50% oil-in-water emulsions. Comparatively, the emulsions formed using extract from non-processed wattle seed were very stable at both 20% and 50% oil levels, especially at pH 7 where the enhanced viscosity of the extract predominated. All extracts had very low foaming capacity, and gelation did not occur in any of the samples even at 10% (w/v) extract concentration.

Influence of heat processing on the bioaccessibility of zinc and iron from cereals and pulses consumed in India

Influence of heat processing on the bioaccessibility of zinc and iron from food grains consumed in India was evaluated. Cereals – rice ( Oryza sativa), finger millet ( Eleusine coracana), sorghum ( Sorghum vulgare), wheat ( Triticum aestivum), and maize ( Zea mays), and pulses – chickpea ( Cicer arietinum) – whole and decorticated, green gram ( Phaseolus aureus) – whole and decorticated, decorticated black gram ( Phaseolus mungo), decorticated red gram ( Cajanus cajan), cowpea ( Vigna catjang), and French bean ( Phaseolus vulgaris) were examined for zinc and iron bioaccessibility by employing an in vitro dialysability procedure. Both pressure-cooking and microwave heating were tested for their influence on mineral bioaccessibility. Zinc bioaccessibility from food grains was considerably reduced upon pressure-cooking, especially in pulses. Among cereals, pressure-cooking decreased zinc bioaccessibility by 63% and 57% in finger millet and rice, respectively. All the pressure-cooked cereals showed similar percent zinc bioaccessibility with the exception of finger millet. Bioaccessibility of zinc from pulses was generally lower as a result of pressure-cooking or microwave heating. The decrease in bioaccessibility of zinc caused by microwave heating ranged from 11.4% in chickpea (whole) to 63% in cowpea. Decrease in zinc bioaccessibility was 48% in pressure-cooked whole chickpea, 45% and 55% in pressure-cooked or microwave-heated whole green gram, 32% and 22% in pressure-cooked or microwave-heated decorticated green gram, and 45% in microwave-heated black gram. Iron bioaccessibility, on the other hand, was significantly enhanced generally from all the food grains studied upon heat treatment. Thus, heat treatment of grains produced contrasting effect on zinc and iron bioaccessibility.

Influence of heat processing and calcium ions on the ability of EDTA to inhibit lipid oxidation in oil-in-water emulsions containing omega-3 fatty acids

The nutritional benefits of ω-3 fatty acids make them excellent candidates as functional food ingredients if problems with oxidative rancidity can be overcome. Oil-in-water emulsions were prepared with 2% salmon oil, stabilized by 0.2% Brij 35 at pH 7. To determine the effects of heating (50–90 °C), ethylenediaminetetraacetic acid (EDTA), and calcium on the oxidative and physical stability of salmon oil-in-water emulsions, particle size, thiobarbituric acid reactive substances (TBARS), and lipid hydroperoxides were measured. The heat-processed emulsions showed no significant difference, in particle size, TBARS or hydroperoxides during storage, from unheated emulsions. Above 2.5 μM, EDTA dramatically decreased lipid oxidation in all samples. Addition of calcium to emulsions containing 7.5 μM EDTA significantly increased both TBARS and hydroperoxide formation when calcium concentrations were 2-fold greater than EDTA concentrations. These results indicate that heat-processed salmon oil-in-water emulsions with high physical and oxidative stability could be produced in the presence of EDTA.

Heat processing of cereals in mash or pellet diets for young pigs

The influence of heat processing (HP) of the cereal on performance and component digestibility in piglets fed on diets offered in mash or pellet form was determined. In trial one, there were eight treatments arranged factorially with two cereals (barley or maize), two HP of the cereal (raw or cooked-flaked) and two forms of the diet (mash or pellets). Piglets were fed on the experimental diets from weaning at 22 to 42 days and then on a common pelleted starter diet until 52 days of age. At 42 days, piglets fed on maize-based diets had better feed conversion ratio (1.20 versus 1.41; P<0.01) than piglets fed on barley-based diets. Heat processing of the cereal improved growth rate at 32 days (0.135 kg per day versus 0.114 kg per day; P<0.05) but no differences were detected thereafter. Pelleting improved feed conversion at all ages ( P<0.01) but not daily gain. At 52 days of age, the only differences remaining were for daily gain ( P<0.05) between maize and barley diets and for feed conversion ( P<0.001) between pellet and mash diets. Coefficient of total tract apparent digestibility of components were higher for maize than for barley-based diets and were improved by both HP of the cereal and pelleting ( P<0.05). In trial two, the coefficient of ileal apparent digestibility of starch, the volatile fatty acid concentration at the caecum and the pH of the gut content were measured in piglets fed on the mash diets tested in trial one. Starch digestibility tended to improve with HP (0.985 versus 0.977; P<0.10). Processed cereals gave lower propionic and butyric acid concentrations and higher caecal pH than raw cereals ( P<0.05). In summary, maize diets gave better performance and digestibility than barley diets; HP of the cereal improved digestibility and growth rate only for the first 10 day post-weaning and pelleting of the diet improved digestibility and feed conversion but not growth rate.

Heat processing of barley and enzyme supplementation of diets for broilers

The influence of heat processing (HP) of barley and enzyme supplementation (ES) of the diet on digestive and performance traits of broilers to 21 d was studied. There were four treatments arranged factorially with two barley-processing treatments (raw or heated), two levels of ES (0 or 500 ppm), and five replicates per treatment. Chicks fed HP barley grew faster than broilers fed raw barley until 8 d of age, but the effect disappeared thereafter. In general, ES improved broiler performance at all ages. Intestinal viscosity was increased by HP of barley (P < or = 0.05) and reduced by ES (P < or = 0.001), and the decrease in viscosity caused by ES was greater for HP than for raw barley diets (HP × ES; P < or = 0.05). Heat processing of barley and ES of the diet improved apparent retention of nutrients (P < or = 0.001). Age affected apparent retention of nutrients differently. For starch and neutral detergent fiber, retention increased linearly with age (P < or = 0.01), but for the remaining nutrients the retention decreased from d 4 to 8 and then increased until d 21 (P < or = 0.001). Also, the beneficial effects of HP on retention of nutrients were more pronounced at younger ages (HP × age; P < or = 0.05). Both HP (P < or = 0.001) and ES (P < or = 0.01) increased liver weight, and enzymes reduced the weights of pancreas (P < or = 0.05) and small intestine (P < or = 0.001). Villus height was improved by HP (P < or = 0.001) and ES (P < or = 0.01), but villus surface area was only improved by enzymes (P < or = 0.01). It was concluded that broiler performance is improved by HP of barley at early ages and by ES of the diet throughout the trial. Also, HP and ES increased apparent retention of nutrients, AMEn of the diet, and villus height.