Nutritional composition, lipid profile and stability, antioxidant activities and sensory evaluation of pasta enriched by linseed flour and linseed oil.

Pasta assortments fortified with high quality foods are a modern nutritional trends. This study, explored the effects of fortification with linseed flour (LF) and linseed oil (LO) on durum wheat pasta characteristics. Wheat flour semolina was replaced with 5%, 10% and 15% of LF or 1%, 2.5% and 5% of LO. Control pasta CP (without LF or LO addition), LF-enriched pasta LFP 5%, LFP 10% and LFP 15% and LO-enriched pasta LOP 1%, LOP 2.5% and LOP 5% was compared for the proteins, fat and phenolic contents and fatty acids (FA) profile. Impact on lipid oxidation and sensory evaluation were also determined. Fortification of pasta with LF improved significantly (p < 0.05) the contents of protein, fat and phenolic compared to CP whereas the enrichment of pasta with LO resulted in a significant increase (p < 0.05) in the content of fat and a significant decrease in protein and phenolic contents. All the formulations decreased the saturated FA percent and increased the polyunsaturated FA percent with enhancement of omega-3 FA content. Antioxidant activity measured by FRAP and DPPH assays was improved after the fortification. For lipid oxidation, the replacement of semolina by LF or LO promoted an increase (p < 0.05) on TBARS values in level-dependent manner. Regarding sensory evaluation, the two types of fortification did not affect the taste; flavor and aroma of cooked pasta, but LOP 5% showed the highest score of the overall acceptability. The results recommended the possibility of producing pasta supplemented with LF or LO (even at a level of 15% and 5% respectively) as a functional food.

Development of a corn flour certified reference material for the accurate determination of zearalenone.

A certified reference material (CRM, KRISS 108-01-002) for zearalenone in corn flour was developed to assure reliable and accurate measurements in testing laboratories. Commercially available corn flour underwent freeze-drying, pulverization, sieving, and homogenization. The final product was packed in amber bottles, approximately 14 g per unit, and preserved at -70 °C. 13C18-Zearalenone was used as an internal standard (IS) for the certification of zearalenone by isotope-dilution liquid chromatography-tandem mass spectrometry (ID-LC‒MS/MS) and for the analysis of α-zearalenol, ß-zearalenol, and zearalanone by LC‒MS/MS. The prepared CRM was sufficiently homogeneous, as the among-unit relative standard deviation for each mycotoxin ranged from 2.2 to 5.7 %. Additionally, the stability of the mycotoxins in the CRM was evaluated under different temperature conditions and scheduled test periods, including storage at -70°C, -20°C, and 4°C and room temperature for up to 12 months, 6 months, and 1 month, respectively. The content of each target mycotoxin in the CRM remained stable throughout the monitoring period at each temperature. Zearalenone content (153.6 ± 8.0 µg/kg) was assigned as the certified value. Meanwhile, the contents of α-zearalenol (1.30 ± 0.17 µg/kg), ß-zearalenol (4.75 ± 0.33 µg/kg), and zearalanone (2.09 ± 0.16 µg/kg) were provided as informative values.

Physicochemical and Morphological Characteristics of Starch and Flour Obtained from Green Banana cv. Raja (Musa paradisiaca cv. Raja) in West Sumatra, Indonesia.

<b>Background and Objective:</b> Banana cv. <i>Raja</i> is widely cultivated in West Sumatra, Indonesia. The physicochemical properties of starch and flour were investigated to determine their functional food prospects in industrial food. <b>Materials and Methods:</b> Starch and flour of banana cv. <i>Raja</i> was characterized using proximate analysis, Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), Fourier-Transform Infrared (FT-IR), X-Ray Fluorescence (XRF) and Rapid Visco-Analyzer (RVA). <b>Results:</b> Banana cv. <i>Raja</i> starch contains 40.73% starch, 17.49% amylose, 55.5% water, 0.66% ash, 0.83% protein and 0.18% fat. The size of the granules is ranging from 20-30 µm in irregular and ellipsoidal-truncated shapes. The structure of crystallinity belongs to the type B while the gelatinization temperature is 74.9°C. Furthermore, the starch composed of 41.06% potassium, 12.85% phosphorus, 12.74% iron, 9.4% calcium and 7.5% magnesium. <b>Conclusion:</b> The morphological and physicochemical starch characteristics of Banana cv. <i>Raja</i> and has similar characteristics with its flour. Meanwhile the swelling power and the solubility value of the flour were higher than the starch. The gelatinization temperatures of starch and flour were 74.9 and 73.4°C, respectively.

Oxysterols in stored powders as potential health hazards.

Cholesterol oxidation products (COPs) have greater biological activity than cholesterol itself. Oxysterols reduce the nutritional value of foods and exhibit a wide range of biological activity, including pro-oxidant, carcinogenic, and cytotoxic properties. The most commonly detected oxysterols in foods are 7α-HC, 7ß-HC, a product of their dehydrogenation 7-KC and α-CE, ß-CE. The main dietary sources of oxysterols are eggs and egg-derived products, thermally processed milk and milk-based products, fried meat. This study aimed to measure the amount of cholesterol oxidation products in milk powder, egg powder and milk-egg powder during 24 months of storage. The changes in the selected oxysterols (determined by gas chromatography) were recorded. In milk powder, after the production process, the amount of cholesterol was 0.2 g 100 g-1 fat and in egg powder it was 3.4 g 100 g-1. After 6 months of storage, the dominant oxysterol in milk and egg powder was 7α-HC and in milk-egg powder it was 7-KC. After the storage period, oxysterols in powdered milk reached 1.81% of total cholesterol.  The most stable cholesterol was in the milk-egg mixture and its oxidation was the slowest. This study showed the presence of COPs in milk powder, egg powder and milk-egg powder and the effect of storage on cholesterol oxidation.

Compositional, thermal and water sorption properties of wheat flour: comparing wild-relatives v/s elite wheat varieties used in current plant breeding.

Plant breeders are focused on creating new varieties of wheat with enhanced yield potential without affecting the quality of wheat for food. In this study, four cultivated varieties of wheat, Triticium asetivum and two wild relatives of wheat; Triticum timopheevii and Triticum urartu were studied. The impact of starch and protein content on structural, thermal and water sorption properties of four main wheat cultivars in comparison to the wild relatives has been studied. Wild relatives of wheat required higher temperatures to initiate starch gelatinisation and amylose/lipid complex formation as compared to the four main wheat cultivars. The carbohydrate region in FTIR indicates structural differences (ordered and unordered starch) in the different varieties of wheat flour. The differentiation between wild relatives and main varieties is at the species level. Both wild relatives showed a noticeable difference in moisture sorption behaviour as compared with the 4 main cultivars, especially in terms of monolayer (M0) and the strength of water binding to the primary sites (C) values.

Rheological characteristics of wheat-chickpea composite flour doughs and effect of Amla powder (Phyllanthus emblica L.) addition on the functional properties of bread.

Bread is a staple food for majority of the people worldwide, but it has a high glycemic effect. Substituting wheat flour partly with chickpea flour and the presence of bran is suggested to improve the glycemic effect of bread; however, the non-gluten substances in wheat flour adversely affect dough rheology. The addition of amla powder was tested on the rheological properties of wheat-chickpea flour composite doughs; also, the physical and sensory qualities of bread made thereof. The results showed that when the level of replacement of refined white flour (WF) or whole wheat flour (WWF) with chickpea flour was increased from 0 to 40%, it significantly affected the rheological properties and functionality of dough. A decreased farinograph water absorption, higher mixing tolerance index (i.e., weakening of dough), decreased resistance to extension, and lower ratio numbers were obtained with some differences between WF and WWF at the higher level of chickpea flour substitution. The addition of amla powder to WF: chickpea flour (60:40) blends reduced the angle of ascending (from 7.0 ± 0.7 to 6.0 ± 0.7) and angel of descending (from 3.2 ± 0.21 to 2.4 ± 0.2), indicating the slight tightening of gluten leading to dough breakdown. The addition of amla powder improved the mixing characteristics of the composite flour doughs, as well as the physical and sensory qualities of the bread. In conclusion, amla powder can help overcome the deleterious impact of adding chickpea flour to WF or WWF for producing good quality pan bread for people with type-2 diabetes.

Effects of dry heat treatment temperature on the structure of wheat flour and starch in vitro digestibility of bread.

The effects of the dry heat treatment (DHT) temperature (20, 50, 100, 150, 200 °C) on the structure of wheat flour and on the texture and in vitro starch digestibility of breads were investigated. X-ray diffraction and FTIR showed that increasing temperatures produced reduction of the hydrated starch structures, increased crystallinity and molecular order of starch chains, and had important effects on the gluten secondary structure. High treatment temperatures produced significant reductions in rapidly digestible starch (RDS) (53.21% at 20 °C, 22.24% at 200 °C), and the slowly digestible starch fraction tended to increase (26.12% at 20 °C, 31.48% at 200 °C). On the other hand, bread hardness showed a significant increase from 11.25 N at 20 °C to 49.53 N at 200 °C, the latter value being similar to that reported for bread crusts. Principal component analysis results showed that the flour and bread characteristics were drastically changed by the DHT, with 100 °C representing a critical temperature. Below 100 °C, breads showed textural characteristics close to that of the control bread, with reduced RDS fractions, while at temperatures above 100 °C, hardness was boosted.

Current calcium fortification experiences: a review.

Low dietary calcium is very common in many populations, contributing to nutritional rickets/osteomalacia in children/adults and increasing the risk of several health problems. Calcium is a nutrient of concern as the recommended nutrient requirements are difficult to meet in the absence of dairy products. The provision of culturally acceptable calcium-fortified foods may improve calcium intake when it is a feasible and cost-effective strategy in a particular setting. This landscape review was conducted in 2019 and describes current calcium fortification efforts and lessons learned from these experiences. Worldwide, the United Kingdom is the only country where calcium fortification of wheat flour is mandatory. It is estimated that this fortified staple ingredient contributes to 13-14% of calcium intake of the British population. Other items voluntary fortified with calcium include maize flour, rice, and water. Current calcium fortification programs may lack qualified personnel/training, clear guidelines on implementation, regulation, monitoring/evaluation, and functional indicators. Also, the cost of calcium premix is high and the target groups may be hard to reach. There is a lack of rigorous evaluation, particularly in settings with multiple micronutrient programs implemented simultaneously, with low quality of the evidence. Further research is needed to assess the impact of calcium fortification programs.

Understanding whole-wheat flour and its effect in breads: A review.

Although the consumption of whole grains, including bread made with whole-wheat flour, is promoted for health benefits and reduced risk for disease and mortality, consumer acceptance, and consumption of some whole-wheat products is low compared to that of white breads. This review focuses on the understanding of whole-wheat flours, both their positive and negative aspects, and how to improve those flours for the production of whole-wheat breads. The review addresses genetic aspects, various milling systems, and pretreatment of bran and germ. The baking process and use of additives and enzymes may also improve product quality to help consumers meet dietary recommendations for daily whole-wheat consumption.

Effects of HMW-GSs on quality related traits in wheat (Triticum aestivum L.) under different water regimes.

Alleles at the Glu-1 loci play important roles in the functional properties of wheat flour. The effects of various high-molecular-weight glutenin subunit (HMW-GS) compositions on quality traits and bread-making properties were evaluated using 235 doubled haploid lines (DHs). The experiment was conducted in a split plot design with two water regimes as the main plot treatment, and DH lines as the subplot treatments. Results showed that the presence of subunit pair 5+10 at the Glu-D1 locus, either alone or in combination with others, appears to provide an improvement in quality and bread-making properties. At the Glu-A1 locus, subunit 1 produced a higher Zeleny sedimentation value (Zel) and stretch area (SA) than subunit 2* when subunits 14+15 and 5+10 were expressed at the Glu-B1 and Glu-D1 loci, and 2* had a positive effect on the maximum dough resistance (Rmax) when subunits 14+15 and 5'+12 were expressed at the Glu-B1 and Glu-D1 loci, respectively. Given subunit 1 at the Glu-A1 locus and 5'+12 at the Glu-D1 locus, the effects of Glu-B1 subunits 14+15 on the tractility (Tra), dough stability time (ST), and dough development time (DT) under the well-watered regime were significantly higher than those of Glu-B1 subunits 13+16. However, 13+16 had a positive effect on SA under the rain-fed regime when subunits 2* and 5+10 were expressed at the Glu-A1 and Glu-D1 loci, respectively. Multiple comparisons analysis revealed that the Zel and Rmax of the six subunits and eight HMW-GS compositions were stable under different water regimes. Overall, subunit compositions 1, 13+16 and 5+10 and 1, 14+15 and 5+10 had higher values for quality traits and bread-baking properties under the two water regimes. These results could play a positive guiding role in selecting and popularizing varieties suitable for production and cultivation in local areas.

Exploitation of Lactic Acid Bacteria and Baker's Yeast as Single or Multiple Starter Cultures of Wheat Flour Dough Enriched with Soy Flour.

Sourdough fermentation presents several advantageous effects in bread making, like improved nutritional quality and increased shelf life. Three types of experiments aimed to evaluate comparatively the efficiency of two Lactobacillus (Lb.) strains, Lb. plantarum ATCC 8014 and Lb. casei ATCC 393, to metabolize different white wheat flour and soybeans flour combinations to compare their efficiency, together with/without Saccharomyces cerevisiae on sourdough fermentation. For this purpose, the viability, pH, organic acids, and secondary metabolites production were investigated, together with the dynamic rheological properties of the sourdough. During sourdough fermentation, LAB presented higher growth, and the pH decreased significantly from above pH 6 at 0 h to values under 4 at 24 h for each experiment. Co-cultures of LAB and yeast produced a higher quantity of lactic acid than single cultures, especially in sourdough enriched with soy-flour. In general, sourdoughs displayed a stable, elastic-like behavior, and the incorporation of soy-flour conferred higher elasticity in comparison with sourdoughs without soy-flour. The higher elasticity of sourdoughs enriched with soy-flour can be attributed to the fact that through frozen storage, soy proteins have better water holding capacity. In conclusion, sourdough supplemented with 10% soy-flour had better rheological properties, increased lactic, acetic, and citric acid production.

Development and validation of high-throughput and low-cost STARP assays for genes underpinning economically important traits in wheat.

KEY MESSAGE: We developed and validated 56 gene-specific semi-thermal asymmetric reverse PCR (STARP) markers for 46 genes of important wheat quality, biotic and abiotic stress resistance, grain yield, and adaptation-related traits for marker-assisted selection in wheat breeding. Development of high-throughput, low-cost, gene-specific molecular markers is important for marker-assisted selection in wheat breeding. In this study, we developed 56 gene-specific semi-thermal asymmetric reverse PCR (STARP) markers for wheat quality, tolerance to biotic and abiotic stresses, grain yield, and adaptation-related traits. The STARP assays were validated by (1) comparison of the assays with corresponding diagnostic STS/CAPS markers on 40 diverse wheat cultivars and (2) characterization of allelic effects based on the phenotypic and genotypic data of three segregating populations and 305 diverse wheat accessions from China and 13 other countries. The STARP assays showed the advantages of high-throughput, accuracy, flexibility, simple assay design, low operational costs, and platform compatibility. The state-of-the-art assays of this study provide a robust and reliable molecular marker toolkit for wheat breeding programs.

Effect of Vacuum Steam Treatment of Hard Red Spring Wheat on Flour Quality and Reduction of Escherichia coli O121 and Salmonella Enteritidis PT 30.

ABSTRACT: Recent outbreaks traced to contaminated flour have created a need in the milling industry for a process that reduces pathogens in wheat while maintaining its functional properties. Vacuum steam treatment is a promising technology for treatment of low-moisture foods. Traditional thermal treatment methods can compromise wheat functionality due to high temperatures; thus, maintaining the functional quality of the wheat protein was critical for this research. The objective of this study was to evaluate the effect of vacuum steam treatment of hard red spring (HRS) wheat kernels on final flour quality and the overall efficacy of vacuum stream treatment for reducing pathogens on HRS wheat kernels. HRS wheat samples were treated with steam under vacuum at 65, 70, 75, and 85°C for 4 and 8 min. Significant changes in dough and baked product functionality were observed for treatments at ≥70°C. Treatment time had no significant effect on the qualities evaluated. After determining that vacuum steam treatment at 65°C best preserved product quality, HRS wheat was inoculated with Escherichia coli O121 and Salmonella Enteritidis PT 30 and processed at 65°C for 0, 2, 4, 6, or 8 min. The treatments achieved a maximum average reduction of 3.57 ± 0.33 log CFU/g for E. coli O121 and 3.21 ± 0.27 log CFU/g for Salmonella. Vacuum steam treatment could be an effective pathogen inactivation method for the flour milling industry.

Mechanism by which ß-glucanase improves the quality of fermented barley flour-based food products.

The enrichment of ß-glucan in barley significantly decreases the quality of dough and baked bread, and ß-glucanase can improve dough and bread quality. Nevertheless, the mechanism by which ß-glucanase improves the quality of fermented barley flour-based products is still poorly understood. The gluten microcosmic structure, molecular structure and yeast gas production capacity were investigated using Fourier transform infrared (FT-IR) spectroscopy, Raman spectroscopy, Scanning electron microscopy (SEM) and F3 rheological fermentation techniques. The results showed that ß-Glucanase can degrade the high-molecular-weight ß-glucan to low-molecular-weight oligosaccharide fragments, which reduces the viscosity of the ß-glucans, promotes cross-linking between the gluten molecules, and indirectly improves the gluten network structure, thereby alleviating the negative effects of ß-glucans. The ß-glucosidase produced during yeast metabolism can further hydrolyse low-molecular-weight oligosaccharides into reducing sugars that can be used by yeast, increasing the carbon sources available to yeasts and the gas production capability of yeasts.

Genomic Prediction and Genome-Wide Association Studies of Flour Yield and Alveograph Quality Traits Using Advanced Winter Wheat Breeding Material.

Use of genetic markers and genomic prediction might improve genetic gain for quality traits in wheat breeding programs. Here, flour yield and Alveograph quality traits were inspected in 635 F6 winter wheat breeding lines from two breeding cycles. Genome-wide association studies revealed single nucleotide polymorphisms (SNPs) on chromosome 5D significantly associated with flour yield, Alveograph P (dough tenacity), and Alveograph W (dough strength). Additionally, SNPs on chromosome 1D were associated with Alveograph P and W, SNPs on chromosome 1B were associated with Alveograph P, and SNPs on chromosome 4A were associated with Alveograph L (dough extensibility). Predictive abilities based on genomic best linear unbiased prediction (GBLUP) models ranged from 0.50 for flour yield to 0.79 for Alveograph W based on a leave-one-out cross-validation strategy. Predictive abilities were negatively affected by smaller training set sizes, lower genetic relationship between lines in training and validation sets, and by genotype-environment (G×E) interactions. Bayesian Power Lasso models and genomic feature models resulted in similar or slightly improved predictions compared to GBLUP models. SNPs with the largest effects can be used for screening large numbers of lines in early generations in breeding programs to select lines that potentially have good quality traits. In later generations, genomic predictions might be used for a more accurate selection of high quality wheat lines.

Certification of Ochratoxin A Reference Materials: Calibration Solutions OTAN-1 and OTAL-1 and a Mycotoxin-Contaminated Rye Flour MYCO-1.

Background: Among the regulated mycotoxins that contaminate global food supplies, ochratoxin A is particularly harmful as a nephrotoxin and suspected carcinogen. Objective: To support global measurement comparability, certified calibration solutions for ochratoxin A and [13C6]-ochratoxin A (OTAN-1 and OTAL-1, respectively) as well as a mycotoxin-contaminated rye flour certified reference material (CRM) known as MYCO-1 were developed. Methods: Quantitative proton NMR was used along with maleic acid as an external standard traceable to the Système international (SI) to measure the concentration of ochratoxin A and [13C6]-ochratoxin A for the calibration solutions. OTAN-1 and OTAL-1 were then used as a pair in double isotope dilution MS to certify the mass fraction of ochratoxin A in MYCO-1. The natural ochratoxin A CRM served as the primary standard for traceable quantitation, while the synthetic [13C6]-ochratoxin A CRM served as the internal standard. Results: The certified mass fraction of ochratoxin A or [13C6]-ochratoxin A in the two mycotoxin calibration solution standards was established to be 11.03 ± 0.32 µg/g (k = 2) for OTAN-1 and 4.89 ± 0.18 µg/g (k = 2) for OTAL-1. The mass fraction of ochratoxin A in the rye flour standard MYCO-1 was certified at 4.05 ± 0.88 µg/kg (k = 2). Conclusions: These CRMs will support regulatory testing as they can be used in the method development, validation, calibration, and QC analysis of ochratoxin A. Highlights: This report highlights the methods used to certify OTAN-1, OTAL-1, and MYCO-1 as well as the challenges associated with producing such materials, which can be applied to a wide variety of other CRMs.

Impacts of short-term germination on the chemical compositions, technological characteristics and nutritional quality of yellow pea and faba bean flours.

In the present study, yellow pea (CDC Amarillo) and faba bean (CDC Snowdrop) seeds were soaked overnight and then germinated in the dark at ambient temperature for 24, 48 and 72 h. During the short-term germination, germination percentages higher than 96.6% were achieved and progressive growth of radicles was observed for both varieties. The soaked and germinated seeds were dried at 55 °C and milled into flours, and their chemical compositions, physicochemical properties and in vitro starch and protein digestibility were systematically examined. Overall, soaking and germination did not noticeably alter the chemical compositions of each flour. The most obvious changes in the physicochemical properties were found in the pasting, emulsifying and foaming properties of the pulse flours. Soaking and 24-h germination greatly enhanced the pasting viscosities of the flours; as the germination proceeded, their viscosities gradually decreased, resulting from the degradation of starch by endogenous amylase(s) during pasting. Germination progressively improved the emulsion activity and stability, foaming capacity and foam stability of both pulse flours. In addition, germination enhanced the in vitro digestibility of starch and protein of the flours; however, the treatment did not improve their in vitro protein digestibility corrected amino acid scores (IV-PDCAAS). Short-term germination of 24-72 h has been demonstrated to be an effective approach to generating pulse flours possessing diverse functional properties and enhanced digestibility of macronutrients.

Análise de sujidades e matérias estranhas em amostras de farinha de trigo tipo 1 comercializadas no município de Campo Mourão, Paraná / Analysis of light filth and strange matters in samples of wheat flour type 1 marketed in Campo Mourão, Paraná

A farinha de trigo é obtida por moagem e suas características são influenciadas por condições a que são submetidas na producão. A contaminação deste produto pode se dar desde a infestação dos grãos de trigo por pragas do campo até o seu ambiente de armazenagem. Com isso torna-se necessário o uso das boas práticas de produção para garantir a qualidade do produto e evitar riscos ao consumidor. Neste sentido, a ANVISA estabelece um limite de tolerância para avaliação de matérias estranhas macroscópicas e microscópicas em alimentos. Este trabalho buscou coletar, analisar e verificar a qualidade de amostras de farinha de trigo tipo 1, afim de verificar sua adequação frente a legislação vigente, tendo como base o método da “Association of Official of Analytical Chemists International” (AOAC). Os resultados mostraram, presença de matérias estranhas, porém dentro dos limites estabelecidos, demonstrando qualidade em relação à presença de sujidades.

Effects of high hydrostatic pressure on quality changes of blends with low-protein wheat and oat flour and derivative foods.

This study aimed to investigate the effect of high hydrostatic pressure (HHP) on the physicochemical characteristics of blended low-protein wheat (LW) and oat flour. Additionally, quality changes in noodles made from blends treated with HHP were investigated. Crude protein and fiber contents of LW were not affected by HHP; however, those of blends were significantly higher than those of LW (p < 0.05). Water-holding capacity (WHC) of blends increased with HHP treatment. The peak viscosity of LW did not differ significantly because of HHP, and the peak and final viscosities of blends increased upon oat flour addition. The hardness, gumminess, chewiness of noodles made using LW improved with the addition of oat flour combined with HHP. The results indicated that the use of blends containing LW and oat flour as well as HHP treatment improved the quality and properties of noodles made using LW.

Cloning and characterization of a novel low-molecular-weight glutenin subunit gene with an unusual molecular structure of Aegilops uniaristata.

Low-molecular-weight glutenin subunits (LMW-GSs) are one of the important factors for the dough processing quality. In this study, a novel LMW-GS, designated LMW-N13, from the wheat relative species Aegilops uniaristata PI 554421 was cloned and characterized. Unlike previously published LMW-GSs, LMW-N13 has a large molecular weight and is the largest LMW-GS published thus far. Sequence alignments demonstrated that LMW-N13 is a LMW-i-type subunit but contains nine cysteine residues which is one more than typical LMW-i-type subunits. In addition, four insertions are present in the repetitive domain that resulted in the large molecular weight. In vitro analysis showed that LMW-N13 could improve the dough quality of different base flours.