101 results about "Egg phospholipids" patented technology

The invention provides an AS01 derivative adjuvant, a preparation method thereof and application of the AS01 derivative adjuvant in preparation of vaccines or anti-tumor drugs, and belongs to the technical field of biological drugs or products. The AS01 derivative adjuvant is prepared from the following components in parts by mass: 0.5 to 1.0 part of cholesterol, 2.0 to 4.0 parts of dioleoyl lecithin and at least one of the following components in parts by mass: 0.1 to 0.2 part of monophosphoryl lipid A, 0.1 to 0.2 part of QS-21, 0.36 to 0.72 part of 3M-052, 0.4 to 0.8 part of muramyl dipeptide and 0.2 to 0.4 part of oligodeoxynucleotide. According to the AS01 derived adjuvant, a novel synergistic adjuvant system is developed by integrating various immunomodulators, so that the immunogenicity of a vaccine is enhanced. The HPV vaccine is prepared on the basis of the AS01 derived adjuvant, so that the immunogenicity can be enhanced, and good anti-tumor activity can be achieved. Therefore, the invention provides a new strategy for immunotherapy of HPV-related tumors.

The invention discloses a hair follicle targeting nano-liposome anti-hair loss and hair growth compound preparation. The compound preparation is prepared from the following raw materials in parts by mass: 70 to 85 parts of bionic liposome carrier and 15 to 30 parts of active ingredients, the bionic liposome carrier is prepared from 50 to 60 parts of egg yolk lecithin, 15 to 20 parts of pH sensitive phospholipid DOPE, 10 to 15 parts of cholesterol and 5 to 10 parts of mesenchymal stem cell exosome membrane protein; the active ingredients comprise 5 to 10 parts of minoxidil, 3 to 5 parts of caffeine, 0.5 to 1 part of miR-218-5p, 1 to 2 parts of biotin and 2 to 3 parts of quercetin; the average particle size of the nano-liposome formed by the bionic liposome carrier is 120-180nm, and the polydispersity index (PDI) is less than or equal to 0.15. The invention relates to the technical field of biological medicines, in particular to a hair follicle-targeted nano-liposome anti-hair loss and hair growth compound preparation, which has the following advantages due to the adoption of the technical scheme: accurate targeted delivery is realized, and the treatment safety and accuracy are improved; intelligent response release is achieved, and the local microenvironment of hair follicles is optimized; the multi-target synergistic effect breaks through the limitation of single treatment.

The invention discloses a lipophilic liposome containing polydeoxyribonucleotide as well as a preparation method and application of the lipophilic liposome. The lipophilic liposome comprises the following components in parts by weight: 0.001-3 parts of polydeoxyribonucleotide; 0.5 to 3 parts of phospholipid; 0.1-2 parts of a cationic emulsifier; 0.1-3 parts of an anionic emulsifier; 40 to 70 parts of polyol; the total amount is 100 parts. According to the invention, an emulsifier system formed by combining lecithin with anionic and cationic compound surfactants is adopted, so that the transdermal absorption of PDRN is facilitated. The lipophilic liposome containing polydeoxyribonucleotide prepared by the invention can effectively resist nuclease degradation, prolong the acting time of PDRN in skin, and improve the skin anti-aging and repairing effects of the liposome. The anti-aging effect of the active ingredients is improved, and the bioavailability of the active ingredients is improved. The cosmetic composition can be widely applied to cosmetic formulas of various dosage forms.

The invention relates to the technical field of nano drug loading, in particular to chlorin e6 composite liposome as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving egg yolk lecithin, cholesterol and chlorin e6 in an organic solvent, carrying out rotary evaporation in a constant-temperature water bath kettle to evaporate the solvent to dryness to form a lipoid film, adding a crocin aqueous solution, carrying out high-shear dispersion on the film, carrying out ultrasonication, and filtering with a 0.22 [mu] m filter membrane to obtain the chlorin e6 composite liposome. The chlorin e6 composite liposome prepared by the invention is high in encapsulation efficiency and good in stability, and has the effects of enhancing tumor cell uptake and killing tumor cells through photodynamic.

The invention relates to a novel camellia saponin derivative feed additive and a preparation method thereof, and belongs to the technical field of feed additives. The modified chitosan is used in the flocculation and impurity removal process for preparing the camellia oleifera saponin derivative, and the modified chitosan reduces the loss of effective components caused by flocculation and impurity removal while ensuring the purity of the camellia oleifera saponin derivative; soybean lecithin and gelatin are used as capsule walls, the camellia saponin derivative and pyropheophorbide-a are wrapped, pyropheophorbide-a can generate singlet oxygen under excitation of visible light, phospholipid bonds are damaged, the camellia saponin derivative is released, and therefore the slow release effect is achieved; the novel camellia saponin derivative feed additive prepared by the invention has good antibacterial property and timeliness.

The invention discloses a drug-loaded protein-phospholipid composite nanoparticle as well as a preparation method and application thereof. The drug-loaded protein-phospholipid composite nanoparticle has a core-shell structure, an inner core comprises silk fibroin and a hydrophilic drug, a shell layer comprises a plurality of phospholipid bimolecular layers which are stacked layer by layer, each phospholipid bimolecular layer comprises soya bean lecithin, a hydrophobic drug is embedded in a hydrophobic region in each phospholipid bimolecular layer, and the hydrophobic drug is embedded in a hydrophobic region in each phospholipid bimolecular layer. Silk fibroin and a hydrophilic drug are embedded in the hydrophilic area, and zein and cholesterol are embedded in the phospholipid bilayer. The drug-loaded protein-phospholipid composite nanoparticle can simultaneously embed a hydrophobic drug and a hydrophilic drug, has the advantages of large drug loading capacity, high bioavailability, good stability, good transdermal permeation effect and the like, has wide application prospects in the fields of transdermal drug delivery preparations and skin care products, and is simple in preparation method, mild in reaction condition and suitable for industrial production. The method is suitable for large-scale industrial production and application.

The invention discloses a calcium liposome for promoting bone health as well as a preparation method and application thereof. The calcium liposome is prepared from the following components in parts by weight: 3 to 7 parts of phospholipid composition, 0.1 to 0.35 part of phytosterol, 1 to 5 parts of insoluble calcium, 1 to 7.5 parts of protein and 0.1 to 1 part of modifier, wherein the phospholipid composition consists of egg yolk lecithin, phosphatidylinositol and distearoyl phosphatidylcholine, and the mass ratio of the egg yolk lecithin to the phosphatidylinositol to the distearoyl phosphatidylcholine is (8-12): (7-11): ( The specific phospholipid composition obtained through screening is combined with phytosterol to serve as a membrane material to wrap a compound of insoluble calcium and protein, so that the dispersity and the stability of the prepared calcium liposome are obviously improved, when the calcium liposome is applied to preparation of calcium soft capsules, the oil leakage phenomenon is obviously reduced, meanwhile, the absorption of an organism to calcium is also enhanced, and the bioavailability of the calcium soft capsules is improved. Particularly, the bone mineral density and the bone calcium content of an organism are obviously increased, healthy development of bones is promoted, and the difficulty and the pain point of insoluble calcium applied to the fields of functional foods, health care products, biological medicines and the like are effectively solved.

The invention provides a multifunctional cisplatin liposome as well as a preparation method and application thereof. The liposome is prepared from epigallocatechin gallate, cis-platinum, zinc peroxide nanoparticles, a distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 long-acting fragment, a distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-oestrone targeting fragment, lecithin and cholesterol, and the particle size of the liposome is 80 to 130 nm. According to the multifunctional liposome disclosed by the invention, the zinc peroxide nanoparticles are introduced, so that the function of slowly releasing oxygen in a tumor microenvironment can be realized, and oxygen deficit is reversed; through targeting fragment modification, an active targeting effect can be realized by combining with an estrogen receptor highly expressed on the surface of a breast cancer cell; the epigallocatechin gallate and cis-platinum are co-loaded, so that the sensitivity of chemotherapeutic drugs can be improved, the toxic and side effects of the chemotherapeutic drugs are reduced, a good breast cancer resisting effect is achieved, and the tumor volume is remarkably reduced.

The invention provides an LPC liposome preparation for brain-targeted delivery of chemotherapeutic drugs as well as a preparation method and application of the LPC liposome preparation. The LPC liposome preparation comprises L-alpha-lysophosphatidylcholine, egg yolk lecithin and cholesterol, and the mass of the L-alpha-lysophosphatidylcholine is 5%-30% of the total mass of the L-alpha-lysophosphatidylcholine, the egg yolk lecithin and the cholesterol. The constructed LPC lipidosome can accelerate endocytosis of the lipidosome in bECs, and the lipidosome is transferred to an endosome recovery way by up-regulating p62, so that the lipidosome is prevented from entering a lysosome degradation way, and the transendocytosis efficiency is remarkably improved.

The invention provides ROS (reactive oxygen species)-responsive cationic liposome-loaded tryptanthrin as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The ROS response cationic liposome loaded tryptanthrin disclosed by the invention comprises an ROS response cationic liposome and tryptanthrin which is entrapped in the ROS response cationic liposome; the ROS response cationic liposome is prepared from the following raw materials: egg yolk lecithin, cholesterol, DSPE-TK-PEG (distearoyl phosphatidylcholine) and DOTAP (dioctyl phthalate). In view of significant increase of intestinal local active oxygen under the ECLS condition, the ROS-responsive cationic liposome is adopted to wrap tryptanthrin, so that tryptanthrin is selectively released in an intestinal injury area, whole body exposure is reduced, and meanwhile, the protection effect on the intestinal tract is enhanced.

The present invention provides a nutritional composition comprising milk phospholipids and egg phospholipids, particularly suitable as a nutritional composition for infants, also as a nutritional support product for pregnant women and as a nutritional supplement for the elderly, in which phosphatidylcholine and sphingomyelin and hence choline are also highly correlated. The invention provides a nutritional composition containing choline with high bioavailability.

The invention discloses a docetaxel-based liposome drug delivery system as well as a preparation method and application thereof, and belongs to the technical field of anti-tumor nano-drugs. The delivery system is composed of an active pharmaceutical ingredient docetaxel and a specific excipient, and comprises egg yolk lecithin, phosphatidylserine, taraxasterol, protamine oligopeptide, a necessary stabilizer and a necessary pH regulator. Through an optimized preparation process, the nano drug delivery system with good stability is successfully constructed. Experiments show that the liposome is high in encapsulation efficiency and has a good application prospect.

This invention relates to the field of asiaticoside technology, and discloses asiaticoside liposome compositions, preparation methods, and applications. Specifically, based on molecular design principles, monoprotonated tertiary amine permeation enhancers and diprotonated tertiary amine permeation enhancers are synthesized. Using egg yolk lecithin, cholesterol, monoprotonated or diprotonated tertiary amine permeation enhancers, and asiaticoside as raw materials, asiaticoside liposome compositions are prepared by thin-film dispersion. The liposome composition exhibits a transdermal permeability of >90% after 48 hours, demonstrating excellent transdermal performance, and also possesses excellent encapsulation and drug loading properties, making it suitable for preparing gel formulations for treating skin lesions.

The invention discloses a soybean protein-phospholipid biomimetic membrane as well as a preparation method and application thereof. The biomimetic membrane is formed by three raw materials of soybean protein, soybean lecithin and a charged polymer through electrostatic interaction and a physical blending method. The soybean 11s protein solution, the lipidosome dispersion liquid and the charged polymer solution are mixed to obtain the mosaic structure biomimetic membrane which is formed by combining various chemical bond crosslinking and electrostatic interaction, and the process conditions are simple, efficient, convenient and environmentally friendly. The biomimetic membrane provided by the invention has a mosaic structure and the characteristics of the raw materials, so that the biomimetic membrane has high ion passing rate, strong ion selection capability, excellent monovalent cation and divalent cation separation capability and certain biodegradability. When the biomimetic membrane is applied to ion separation, the separation capacity of ions with different valence states can be effectively improved, the separation effect of sodium ions and magnesium ions is optimal, and the separation factor is as high as 21.28 and is 54.6 times that of a biomimetic membrane without charges.

The present invention provides a nutritional composition especially well suited as an infant nutritional composition, but also as a nutritional support product for pregnant women and as senior nutritional supplement, where phosphatidylcholine and sphingomyelin, and thus choline, also are highly relevant. The present invention provides a nutritional composition which contains choline of high bio-availability.

The application relates to the field of biological medicine, and discloses ascorbyl palmitate nanoliposomes capable of inducing tumor cell oncosis, a preparation method and application thereof. The nanoliposomes are constructed by inserting ascorbyl palmitate as a lipid component into a phospholipid bilayer, and are prepared by a film dispersion method from lecithin, cholesterol, pegylated phospholipid and ascorbyl palmitate. The average particle size of the nanoliposomes is 80-150 nm, the encapsulation rate is high, and the stability is good. The nanoliposomes can be administered through a tail vein, utilize the specific hydrolysis of matrix metalloproteinase MMP-2 which is highly expressed by tumor cells to hydrolyze ascorbyl palmitate, release palmitic acid and ascorbic acid, thereby inducing Caspase-independent oncosis of tumor cells, effectively overcoming apoptosis-related drug resistance, and reversing a tumor immunosuppressive microenvironment. The nanoliposomes avoid the cumulative toxicity of metal-type inducers, and provide a new safe and effective option for reversing chemotherapy resistance.

The invention discloses a stable vitamin K1 micelle solution and a preparation method thereof, the vitamin K1 micelle solution comprises 0.75%-1.25% of vitamin K1, 5%-6% of egg yolk lecithin or soybean lecithin, 5.5%-6.5% of polyethylene glycol 1000 vitamin E succinate, a pH regulator and water for injection, and the pH value of the micelle solution is 4.5-6.5. According to the invention, phospholipid and polyethylene glycol 1000 vitamin E succinate (TPGS) are selected as emulsifiers, so that the dissolution of vitamin K1 is solved, the degradation of vitamin K1 is avoided, and the stability of the product is improved; according to the invention, the polyethylene glycol 1000 vitamin E succinate (TPGS) is used, the tocopherol structure of the TPGS has a relatively strong antioxidant function, the content of related impurities of the vitamin K1 is remarkably reduced, and the vitamin K1 has better stability; by controlling the particle size of the micelle to be 65-80 nm, the obtained solution is milky white to faint yellow, and the influence of illumination on the stability of the product is avoided.

A process for producing a triglyceride-phospholipid concentrate is provided, the process comprising: providing an egg yolk lecithin solution from the extraction of an egg yolk material with a liquid extractant, mixing the egg yolk lecithin solution with an OPO triglyceride carrier comprising 1,3-dioleoyl-2-palmitoylglycerol and having a ratio of palmitic acid in the 2-position of the triglycerides relative to the total palmitic acid in the triglycerides (SN2-C16:0) of at least 50%, and a C52 triglyceride content of at least 42 wt.%, and separating the liquid extractant from the mixture to produce a triglyceride- phospholipid concentrate comprising the OPO triglyceride carrier and egg lecithin.

The invention provides a transdermal glabridin nano composition as well as a preparation method and application thereof. The transdermal glabridin nano composition is prepared from oil-phase raw materials: glabridin, soya bean lecithin, an emulsifier and butanediol; the water-phase raw materials comprise p-hydroxyacetophenone, an emulsifier, butanediol, bis (lauramide glutamine) sodium lysine and water. According to the glabridin liposome and the preparation method thereof, the stability of the glabridin liposome, especially the stability at a relatively high temperature, is effectively improved by selecting the proper emulsifier, the storage difficulty of related cosmetics and medical products is reduced, and a good foundation is provided for wider application of the related cosmetics and medical products.

The invention belongs to the technical field of biological medicine preparation, and particularly discloses a preparation method of a curcumin-butylphthalide-phospholipid complex, which comprises the following steps: dissolving curcumin, butylphthalide and lecithin in an organic solvent A, carrying out mixed reaction in a water bath, removing the organic solvent A after the mixed reaction is finished, and purifying by adopting an organic solvent B to obtain the curcumin-butylphthalide-phospholipid complex. And removing the organic solvent B, so as to obtain the curcumin synergistic butylphthalide phospholipid compound. The invention relates to a preparation method of nanoparticles of a curcumin synergistic butylphthalide phospholipid compound, which comprises the following steps: dissolving the curcumin synergistic butylphthalide phospholipid compound in deionized water, oscillating in a water bath to fully dissolve, and filtering through a microfiltration membrane to obtain the nanoparticles of the curcumin synergistic butylphthalide phospholipid compound. Through the synergistic effect of curcumin and butylphthalide, the treatment effect on nervous system diseases is remarkably improved, and a new thought and strategy are provided for treatment of nervous system diseases.

The application belongs to the field of biological medicine, and discloses a nano-lipid preparation for loading perfluorohexane and an antioxidant, wherein the nano-lipid preparation is a core-shell structure, the core-shell structure takes a phospholipid liposome membrane layer as an outer shell, and perfluorohexane and the antioxidant loaded in the outer shell serve as an inner core; the phospholipid liposome membrane layer is made of hydrogenated lecithin, distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000 and cholesterol; and the application discloses an application of the nano-lipid preparation for loading perfluorohexane and the antioxidant in preparation of a medicine for treating myocardial infarction. The nano-lipid preparation has a suitable particle size, can be passively targeted and enriched to a myocardial infarction site, is stable, has a high encapsulation efficiency, and has a simple administration mode; after the lipid preparation is taken by myocardial cells, oxygen and the antioxidant are released, the oxygen can relieve an anoxic condition of a myocardial ischemia site, and the antioxidant can reduce ROS damage, both of which play a synergistic role, improve the survival rate of ischemic myocardial cells, and improve heart function.

The invention provides a high-stability blueberry anthocyanin polycystic liposome and a preparation method of the high-stability blueberry anthocyanin polycystic liposome. The preparation raw materials comprise 5 to 12 parts of a pH response type phospholipid derivative, 3 to 8 parts of mesoporous silica-polydopamine hybrid nanoparticles, 28 to 42 parts of egg yolk lecithin, 8 to 12 parts of cholesterol, 5 to 9 parts of glyceryl trioleate, 2 to 5 parts of vitamin E succinate, 10 to 18 parts of a citric acid-malic acid buffer solution, 45 to 85 parts of trehalose, 16 to 32 parts of a Tween 80-Arabic gum compound, 4 to 8 parts of chitosan quaternary ammonium salt and 5 to 13 parts of polyethylene glycol-2000. 160-280 parts of ultrapure water and 6-22 parts of blueberry anthocyanin. The blueberry anthocyanin polycystic liposome disclosed by the invention has high encapsulation efficiency, excellent photo-thermal stability and long-term storage stability.

The invention relates to the field of biological medicine, and provides an HDAC inhibitor coated liposome degradation agent, which comprises hydrogenated lecithin, cholesterol, an HDAC inhibitor, targeted EGFR drug modified functional phospholipid and E3 ligase ligand modified functional phospholipid. The invention further provides a preparation method of the liposome degradation agent wrapping the HDAC inhibitor and application of the liposome degradation agent in preparation of antitumor drugs. The EGFR-targeted liposome degradation agent is combined with the HDAC inhibitor, malignant proliferation of the osimertinib-resistant non-small cell lung cancer is synergistically overcome from multiple channels, and a new treatment thought is expected to be provided for clinical osimertinib-resistant lung cancer patients.

The invention relates to the technical field of madecassoside, and discloses a madecassoside liposome composition as well as a preparation method and application thereof, specifically, based on a molecular design principle, a monoprotonated tertiary amine penetration enhancer and a diprotonated tertiary amine penetration enhancer are synthesized; yolk lecithin, cholesterol, a monoprotonated tertiary amine penetration enhancer or a diprotonated tertiary amine penetration enhancer and madecassoside are used as raw materials, the madecassoside lipidosome composition is prepared through a film dispersion method, and the transdermal rate of the lipidosome composition after 48 hours is gt; according to the present invention, the prepared hydrogel has excellent transdermal performance, excellent encapsulation performance and excellent drug loading performance, and can be used for preparing a gel preparation for treating skin injury.

A novel liposome-targeted delivery cream containing ectoine and ceramide and its application, mainly used for relieving skin problems such as skin sensitivity, dermatitis, eczema, etc.; the novel liposome combines soybean lecithin, cholesterol, polyethylene glycol-targeting ligand, and the targeting ligand is ectoine to prepare a targeted liposome capable of simultaneously connecting and encapsulating hydrophilic ectoine and hydrophobic ceramide; further added to the cream matrix to develop a novel liposome-targeted delivery cream of compound ectoine combined with ceramide; it has a phospholipid bilayer and is self-closed in structure, can encapsulate ceramide inside the liposome, has good stability, and at the same time stably connects and modifies water-soluble ectoine to prevent its rapid degradation and reduce the drug release rate. Adding it to the cream matrix and directly applying it to the required skin area can achieve targeted transport of the drug in the skin tissue.

The invention discloses a vitamin E-phytosterol oleate composite liposome as well as a preparation method and application thereof, the composite liposome is prepared by taking soybean phospholipid, phytosterol oleate and vitamin E as raw materials and adopting a film ultrasonic method, and the mass ratio of the soybean phospholipid to the phytosterol oleate to the vitamin E is 20: 1: 1. Soybean lecithin and phytosterol oleate are used as main membrane materials to embed vitamin E, the vitamin E-phytosterol oleate composite liposome is constructed, the doping of phytosterol oleate improves the ability of vitamin E to remove DPPH free radicals, and the phytosterol oleate and vitamin E in the liposome play a synergistic anti-oxidation role; by doping the phytosterol oleate, the lipid peroxidation resistance of the vitamin E can be improved; and the reduction capability of the vitamin E is improved by doping the phytosterol oleate. When the additive is applied to the fried instant noodles, the storage stability of the fried instant noodles is effectively improved.

A lipid nanoparticle (liposome) comprising: (i) an outer shell comprising a lipid formulation; and (ii) one or more therapeutic agents which prevent apoptosis of a target cell is provided. The therape

The invention provides a niclosamide and chlorine e6 co-loaded liposome. The niclosamide and chlorine e6 co-loaded liposome is prepared from the following components in parts by weight: 150 to 250 parts of egg yolk lecithin, 5 to 15 parts of cholesterol, 1 to 5 parts of niclosamide, 1 to 5 parts of chlorine e6, 15 to 25 parts of DSPE-SS-MPEG2K and 5 to 20 parts of polyvinyl alcohol. The preparation method is simple to operate and mild in reaction condition. The prepared lipidosome is uniform in particle size, good in water solubility, good in biological safety and high in drug loading capacity, due to introduction of DSPE-SS-MPEG2K, the lipidosome has the glutathione (GSH) response characteristic, not only keeps stable in a normal physiological environment, but also can trigger disulfide bond breakage under the condition of high-concentration GSH in a tumor microenvironment, fixed-point quick release of drugs is achieved, and the application prospect is wide. The breast cancer treatment capability is synergistically enhanced.

The invention discloses a drug-loaded liposome constructed on the basis of saikoside D, and a preparation method and application of the drug-loaded liposome. The liposome comprises soybean lecithin, saikoside D, phosphatidic acid, an active drug and poloxamer 407. According to the invention, the saikosaponin D with antitumor activity is used for completely replacing cholesterol in the traditional liposome, so that the potential health risk of the cholesterol is avoided, the integration of medicines and auxiliary medicines is realized, and the saikosaponin D and entrapped medicines can generate a synergistic antitumor effect. The modification of the poloxamer 407 endows the liposome with the long circulation characteristic. The preparation method adopts a film dispersion-ultrasonic method, and is simple and convenient to operate. The prepared lipidosome is small in particle size, narrow in distribution and good in stability, the drug loading capacity is about 1.5-7.0%, the tumor cell inhibition effect superior to that of traditional cholesterol lipidosome and free drugs is shown in vitro, and the lipidosome has wide application prospects in the aspect of preparation of anti-tumor drugs.

The application relates to the technical field of pharmacy, in particular to a kind of lipid nanoparticles and a preparation method and application thereof.The lipid nanoparticles include nano-phospholipid disc and IMB-AL0912;The nano-phospholipid disc includes lung mucus component phospholipid, negative electric phospholipid, yolk lecithin and 4F peptide;The mass ratio of 4F peptide and lung mucus component phospholipid is 1: (5-10) in terms of mass ratio.A new kind of lipid nanoparticle is obtained by improving nano-phospholipid disc loading polymyxin antibiotic IMB-AL0912, the lipid nanoparticle has the effect of significantly reducing cell damage and inflammatory factor expression caused by LPS stimulation, has a significantly better prevention and treatment effect on bacterial infection and sepsis, is lower in toxicity, and has a wider clinical application range.