acyl coa dehydrogenases


Summary: Enzymes that catalyze the first step in the beta-oxidation of FATTY ACIDS.

Top Publications

  1. Gregersen N, Bross P, Andresen B. Genetic defects in fatty acid beta-oxidation and acyl-CoA dehydrogenases. Molecular pathogenesis and genotype-phenotype relationships. Eur J Biochem. 2004;271:470-82 pubmed
  2. Ghisla S, Thorpe C. Acyl-CoA dehydrogenases. A mechanistic overview. Eur J Biochem. 2004;271:494-508 pubmed
    ..These views are also discussed in the context of the accompanying paper on the three-dimensional properties of acyl-CoA dehydrogenases. ..
  3. Maggio Hall L, Lyne P, Wolff J, Keller N. A single acyl-CoA dehydrogenase is required for catabolism of isoleucine, valine and short-chain fatty acids in Aspergillus nidulans. Fungal Genet Biol. 2008;45:180-9 pubmed publisher
  4. KIM J, Miura R. Acyl-CoA dehydrogenases and acyl-CoA oxidases. Structural basis for mechanistic similarities and differences. Eur J Biochem. 2004;271:483-93 pubmed
    ..In addition, the structure of peroxisomal acyl-CoA oxidase II from rat liver is compared to that of medium chain acyl-CoA dehydrogenase, and the structural basis for their different oxidative half reactions is discussed. ..
  5. Ye X, Ji C, Zhou C, Zeng L, Gu S, Ying K, et al. Cloning and characterization of a human cDNA ACAD10 mapped to chromosome 12q24.1. Mol Biol Rep. 2004;31:191-5 pubmed
    ..RT-PCR result shows that it is widely expressed in human tissues, especially high in liver, kidney, pancreas and spleen. It is hypothesized that this is a novel member of ACADs family. ..
  6. Hanley P, Gopalan K, Lareau R, Srivastava D, von Meltzer M, Daut J. Beta-oxidation of 5-hydroxydecanoate, a putative blocker of mitochondrial ATP-sensitive potassium channels. J Physiol. 2003;547:387-93 pubmed
    ..Our results open the possibility that beta-oxidation of 5-HD or metabolic intermediates of 5-HD may be responsible for the inhibitory effects of 5-HD on preconditioning of the heart. ..
  7. Matsubara Y, Kraus J, Ozasa H, Glassberg R, Finocchiaro G, Ikeda Y, et al. Molecular cloning and nucleotide sequence of cDNA encoding the entire precursor of rat liver medium chain acyl coenzyme A dehydrogenase. J Biol Chem. 1987;262:10104-8 pubmed
    ..The sequencing of other homologous acyl-CoA dehydrogenases will be informative in this regard. ..
  8. Kim J, Wang M, Paschke R. Crystal structures of medium-chain acyl-CoA dehydrogenase from pig liver mitochondria with and without substrate. Proc Natl Acad Sci U S A. 1993;90:7523-7 pubmed
    ..Lys-304, the prevalent mutation site found in patients with medium-chain acyl-CoA dehydrogenase deficiency, is located approximately 20 A away from the active site of the enzyme. ..
  9. Pontoizeau C, Habarou F, Brassier A, Veauville Merllié A, Grisel C, Arnoux J, et al. Hyperprolinemia in Type 2 Glutaric Aciduria and MADD-Like Profiles. JIMD Rep. 2016;27:39-45 pubmed publisher
    ..Analysis of our full cohort of >50,000 samples from >30,000 patients suggests that the proline/alanine ratio may be a good marker of MAD deficiency and could contribute to a more effective management of the treatable forms. ..

More Information


  1. Yang B, Ding J, Zhou C, Dimachkie M, Sweetman L, Dasouki M, et al. Identification of a novel mutation in patients with medium-chain acyl-CoA dehydrogenase deficiency. Mol Genet Metab. 2000;69:259-62 pubmed
    ..Sequence analysis revealed a novel mutation G617T in exon 8 resulting in an arginine to leucine substitution at codon 206 (R206L). Both patients were compound heterozygous for this G617T and the common mutation A985G. ..
  2. Andresen B, Bross P, Udvari S, Kirk J, Gray G, Kmoch S, et al. The molecular basis of medium-chain acyl-CoA dehydrogenase (MCAD) deficiency in compound heterozygous patients: is there correlation between genotype and phenotype?. Hum Mol Genet. 1997;6:695-707 pubmed
    ..Different mutations may contribute with different susceptibilities for disease precipitation, when the patient is subjected to metabolic stress, but other genetic and environmental factors may play an equally important role. ..
  3. Wood P, Kelly Kurtz C, Hinsdale M, Hamm D, Rhead W. Lessons learned from the mouse model of short-chain acyl-CoA dehydrogenase deficiency. Adv Exp Med Biol. 1999;466:395-402 pubmed
  4. Knoll A, Bessoule J, Sargueil F, Cassagne C. Dehydration of 3-hydroxyacyl-CoA in brain very-long-chain fatty acid synthesis. Neurochem Int. 1999;34:255-67 pubmed
    ..These results demonstrated that 3-hydroxyacyl-CoAs entered the elongating complex exclusively at the level of the dehydrase and not of the condensing enzyme. ..
  5. Labinskyy V, Bellomo M, Chandler M, Young M, Lionetti V, Qanud K, et al. Chronic activation of peroxisome proliferator-activated receptor-alpha with fenofibrate prevents alterations in cardiac metabolic phenotype without changing the onset of decompensation in pacing-induced heart failure. J Pharmacol Exp Ther. 2007;321:165-71 pubmed
    ..S. versus control). Thus, preventing changes in myocardial substrate metabolism in the failing heart causes a modest improvement of cardiac function during the progression of the disease, with no effects on the onset of decompensation. ..
  6. Bross P, Engst S, Strauss A, Kelly D, Rasched I, Ghisla S. Characterization of wild-type and an active site mutant of human medium chain acyl-CoA dehydrogenase after expression in Escherichia coli. J Biol Chem. 1990;265:7116-9 pubmed
    ..Properties of the mutant enzyme suggest that the Glu376----Gln change specifically affects substrate binding. These results prove that Glu376 plays an important role in the initial step of dehydrogenation catalysis. ..
  7. Mayatepek E, Ferdinandusse S, Meissner T, Wanders R. Analysis of cysteinyl leukotrienes and their metabolites in bile of patients with peroxisomal or mitochondrial beta-oxidation defects. Clin Chim Acta. 2004;345:89-92 pubmed
    ..g. contributing to liver injury. In addition, our data confirm that the beta-oxidation of cysteinyl LTs in vivo occurs in peroxisomes and not in mitochondria. ..
  8. Förster Fromme K, Chattopadhyay A, Jendrossek D. Biochemical characterization of AtuD from Pseudomonas aeruginosa, the first member of a new subgroup of acyl-CoA dehydrogenases with specificity for citronellyl-CoA. Microbiology. 2008;154:789-96 pubmed publisher
    ..Potential other terpenoid-CoA dehydrogenases were found in the genomes of Pseudomonas citronellolis, Marinobacter aquaeolei and Hahella chejuensis but were absent in non-acyclic terpene-utilizing bacteria. ..
  9. Bzduch V, Behulova D, Lehnert W, Fabriciova K, Kozak L, Salingova A, et al. Metabolic cause of Reye-like syndrome. Bratisl Lek Listy. 2001;102:427-9 pubmed
    ..This highly sophisticated method can detect most of these disorders from dry blood spots disregarding the symptoms and type of mutation. ..
  10. Grice A, Peck T. Multiple acyl-CoA dehydrogenase deficiency: a rare cause of acidosis with an increased anion gap. Br J Anaesth. 2001;86:437-41 pubmed
    ..The pathophysiology of this condition is discussed along with potential treatment options. ..
  11. Erb T, Brecht V, Fuchs G, Muller M, Alber B. Carboxylation mechanism and stereochemistry of crotonyl-CoA carboxylase/reductase, a carboxylating enoyl-thioester reductase. Proc Natl Acad Sci U S A. 2009;106:8871-6 pubmed publisher
    ..This may be useful for protein engineering of enoyl reductases and their application in biocatalysis. ..
  12. Hinsdale M, Kelly C, Wood P. Null allele at Bcd-1 locus in BALB/cByJ mice is due to a deletion in the short-chain acyl-CoA dehydrogenase gene and results in missplicing of mRNA. Genomics. 1993;16:605-11 pubmed
    ..Both abnormal transcripts have aberrant stop codons. These results demonstrate the molecular basis of SCAD deficiency in this unique mouse model. ..
  13. Birkebaek N, Simonsen H, Gregersen N. Hypoglycaemia and elevated urine ethylmalonic acid in a child homozygous for the short-chain acyl-CoA dehydrogenase 625G > A gene variation. Acta Paediatr. 2002;91:480-2 pubmed
    ..Furthermore, gluconeogenesis might be compromised owing to lack of reducing equivalents from the oxidation of short-chain fatty acids in the fasting or stressed state, thus contributing to the predisposition for fasting hypoglycaemia. ..
  14. Wang S, Fernhoff P, Hannon W, Khoury M. Medium chain acyl-CoA dehydrogenase deficiency human genome epidemiology review. Genet Med. 1999;1:332-9 pubmed
    ..State programs are beginning to offer screening in newborns for MCADD using MS/MS. In addition, a private company currently offers voluntary supplemental newborn screening for MCADD to birthing centers. ..
  15. Wilson C, Champion M, Collins J, Clayton P, Leonard J. Outcome of medium chain acyl-CoA dehydrogenase deficiency after diagnosis. Arch Dis Child. 1999;80:459-62 pubmed
    ..Undiagnosed, MCAD deficiency results in considerable mortality and morbidity. However, current management improves outcome, supporting the view that the disorder should be included in newborn screening programmes. ..
  16. Inagaki T, Ohishi N, Tsukagoshi N, Udaka S, Ghisla S, Yagi K. Structurally different rat liver medium-chain acyl CoA dehydrogenases directed by complementary DNAs differing in their 5'-region. Biochim Biophys Acta. 1991;1077:285-90 pubmed
    ..The purified protein (approximately 40 kDa) directed by pRMCADm-6 did not contain FAD. Thus, the deletion of the NH2-terminal portion of the apoprotein to some extent results in its inability to combine with FAD. ..
  17. Gregersen N, Winter V, Corydon M, Corydon T, Rinaldo P, Ribes A, et al. Identification of four new mutations in the short-chain acyl-CoA dehydrogenase (SCAD) gene in two patients: one of the variant alleles, 511C-->T, is present at an unexpectedly high frequency in the general population, as was the case for 625G-->. Hum Mol Genet. 1998;7:619-27 pubmed
  18. Reichmann H, Maltese W, DeVivo D. Enzymes of fatty acid beta-oxidation in developing brain. J Neurochem. 1988;51:339-44 pubmed
  19. Reich Slotky R, Kabbash C, Della Latta P, Blanchard J, Feinmark S, Freeman S, et al. Gemfibrozil inhibits Legionella pneumophila and Mycobacterium tuberculosis enoyl coenzyme A reductases and blocks intracellular growth of these bacteria in macrophages. J Bacteriol. 2009;191:5262-71 pubmed publisher
    ..coli strain sensitive to GFZ at the nonpermissive temperature. GFZ noncompetitively inhibited this L. pneumophila FabI homolog, as well as M. tuberculosis InhA and E. coli FabI. ..
  20. Arent S, Pye V, Henriksen A. Structure and function of plant acyl-CoA oxidases. Plant Physiol Biochem. 2008;46:292-301 pubmed publisher
    ..Structure and sequence comparisons are made with a focus on the enzyme's mechanistic means to control electron transfer paths, reactivity towards molecular oxygen, and spatial and architectural requirements for substrate discrimination. ..
  21. Rhead W, Amendt B, Fritchman K, Felts S. Dicarboxylic aciduria: deficient [1-14C]octanoate oxidation and medium-chain acyl-CoA dehydrogenase in fibroblasts. Science. 1983;221:73-5 pubmed
    ..These data confirm that dicarboxylic aciduria is caused by an enzyme defect in the beta-oxidation cycle. ..
  22. Okun J, Kölker S, Schulze A, Kohlmüller D, Olgemöller K, Lindner M, et al. A method for quantitative acylcarnitine profiling in human skin fibroblasts using unlabelled palmitic acid: diagnosis of fatty acid oxidation disorders and differentiation between biochemical phenotypes of MCAD deficiency. Biochim Biophys Acta. 2002;1584:91-8 pubmed
    ..In conclusion, this novel technique is a powerful tool for the investigation of fatty acid oxidation disorders under standardized conditions in fibroblasts. ..
  23. Zhang J, Zhang W, Zou D, Chen G, Wan T, Zhang M, et al. Cloning and functional characterization of ACAD-9, a novel member of human acyl-CoA dehydrogenase family. Biochem Biophys Res Commun. 2002;297:1033-42 pubmed
    ..Enzymatic assay proved that the recombinant ACAD-9 protein has the dehydrogenase activity on palmitoyl-coenzyme A (C16:0) and stearoyl-coenzyme A (C18:0). Our results indicate that ACAD-9 is a novel member of ACADs. ..
  24. Telford E, Moynihan L, Markham A, Lench N. Isolation and characterisation of a cDNA encoding the precursor for a novel member of the acyl-CoA dehydrogenase gene family. Biochim Biophys Acta. 1999;1446:371-6 pubmed
    ..The cDNA contains an open reading frame of 1248 nucleotides encoding a predicted 415-amino-acid peptide, and shares considerable sequence similarity with other members of the ACD family. ..
  25. Liu H, Reynolds K. Precursor supply for polyketide biosynthesis: the role of crotonyl-CoA reductase. Metab Eng. 2001;3:40-8 pubmed
    ..cinnamonensis indicate the presence of an additional coenzyme B(12)-dependent mutase linking branched and straight-chain C(4) compounds by a new pathway. ..
  26. Willard J, Vicanek C, Battaile K, Van Veldhoven P, Fauq A, Rozen R, et al. Cloning of a cDNA for short/branched chain acyl-Coenzyme A dehydrogenase from rat and characterization of its tissue expression and substrate specificity. Arch Biochem Biophys. 1996;331:127-33 pubmed
    ..Northern blotting of mRNA shows ubiquitous tissue expression of both the rat and human enzyme. Further study of these enzymes will be helpful in understanding structure/function relationships in this gene family. ..
  27. Tanaka K, Yokota I, Coates P, Strauss A, Kelly D, Zhang Z, et al. Mutations in the medium chain acyl-CoA dehydrogenase (MCAD) gene. Hum Mutat. 1992;1:271-9 pubmed
    ..In addition to tabulating the mutations, the acyl-CoA dehydrogenase gene family, the structure of the MCAD gene and the evolution of 985A-->G mutation are briefly discussed. ..
  28. Turpin B, Tobias J. Perioperative management of a child with short-chain acyl-CoA dehydrogenase deficiency. Paediatr Anaesth. 2005;15:771-7 pubmed
    ..We present a 23-month-old girl with SCAD deficiency, who required posterior fossa decompression for type 1 Chiari malformation. The potential perioperative implications of SCAD deficiency are reviewed. ..
  29. Scholte H, Van Coster R, de Jonge P, Poorthuis B, Jeneson J, Andresen B, et al. Myopathy in very-long-chain acyl-CoA dehydrogenase deficiency: clinical and biochemical differences with the fatal cardiac phenotype. Neuromuscul Disord. 1999;9:313-9 pubmed
    ..31P-nuclear magnetic resonance spectroscopy of muscle showed a normal oxidative phosphorylation as assessed by phosphocreatine recovery, but a significant increase in pH and in Pi/ATP ratio. ..
  30. Song W, Qin Y, Saito M, Shirai T, Pujol F, Kastaniotis A, et al. Characterization of two cotton cDNAs encoding trans-2-enoyl-CoA reductase reveals a putative novel NADPH-binding motif. J Exp Bot. 2009;60:1839-48 pubmed publisher
    ..The data suggest that both GhECR genes encode functional enzymes harbouring non-classical NADPH-binding sites at their C-termini, and are involved in fatty acid elongation during cotton fibre development. ..
  31. Wallace K, Bao Z, Dai H, DiGate R, Schuler G, Speedie M, et al. Purification of crotonyl-CoA reductase from Streptomyces collinus and cloning, sequencing and expression of the corresponding gene in Escherichia coli. Eur J Biochem. 1995;233:954-62 pubmed
    ..coli and the induced crotonyl-CoA reductase was purified tenfold and shown to have similar steady-state kinetics and electrophoretic mobility on sodium dodecyl sulfate/polyacrylamide to the native protein. ..
  32. Matsubara Y, Kraus J, Yang Feng T, Francke U, Rosenberg L, Tanaka K. Molecular cloning of cDNAs encoding rat and human medium-chain acyl-CoA dehydrogenase and assignment of the gene to human chromosome 1. Proc Natl Acad Sci U S A. 1986;83:6543-7 pubmed
    ..Direct screening of cDNA libraries using a highly enriched mRNA to generate a probe, as demonstrated in this study, may provide the most rapid and convenient approach to cDNA cloning of low-abundance mRNAs. ..
  33. Kelly D, Whelan A, Ogden M, Alpers R, Zhang Z, Bellus G, et al. Molecular characterization of inherited medium-chain acyl-CoA dehydrogenase deficiency. Proc Natl Acad Sci U S A. 1990;87:9236-40 pubmed
    ..Therefore, the molecular characterization of this family revealed a crucial point mutation in the MCAD gene and an unusual abnormality in MCAD pre-mRNA splicing. ..
  34. Adeva Andany M, Carneiro Freire N, Seco Filgueira M, Fernández Fernández C, Mouriño Bayolo D. Mitochondrial β-oxidation of saturated fatty acids in humans. Mitochondrion. 2018;: pubmed publisher
    ..Accumulation of non-oxidized fatty acids promotes their conjugation with glycine and l-carnitine and alternate ways of oxidation, such as ω-oxidation. ..
  35. Gregersen N, Andresen B, Corydon M, Corydon T, Olsen R, Bolund L, et al. Mutation analysis in mitochondrial fatty acid oxidation defects: Exemplified by acyl-CoA dehydrogenase deficiencies, with special focus on genotype-phenotype relationship. Hum Mutat. 2001;18:169-89 pubmed
    ..However, it remains to be seen to what extent mutation analysis will be used for diagnosis of fatty acid oxidation defects and other metabolic disorders. ..
  36. Lamm T, Kohls T, Saenger A, Stankovich M. Comparison of ligand polarization and enzyme activation in medium- and short-chain acyl-coenzyme A dehydrogenase-novel analog complexes. Arch Biochem Biophys. 2003;409:251-61 pubmed
    ..Such data provide the first indication of variations in the activation mechanism of these enzymes, which were thought to be comparable in both structure and function. ..
  37. Farag E, Argalious M, Narouze S, DeBoer G, Tome J. The anesthetic management of ventricular septal defect (VSD) repair in a child with mitochondrial cytopathy. Can J Anaesth. 2002;49:958-62 pubmed
    ..The anesthetic management of patients with any mitochondrial disease requires normoglycemia, normothermia and the avoidance of metabolic stress in order to preserve energy production by the diseased mitochondria. ..
  38. Han L, Reynolds K. A novel alternate anaplerotic pathway to the glyoxylate cycle in streptomycetes. J Bacteriol. 1997;179:5157-64 pubmed
    ..These results suggested that both ccr and meaA are involved in a novel pathway for the growth of S. collinus when acetate is its sole carbon source...
  39. Swigonova Z, Mohsen A, Vockley J. Acyl-CoA dehydrogenases: Dynamic history of protein family evolution. J Mol Evol. 2009;69:176-93 pubmed publisher
  40. Andresen B, Dobrowolski S, O Reilly L, Muenzer J, McCandless S, Frazier D, et al. Medium-chain acyl-CoA dehydrogenase (MCAD) mutations identified by MS/MS-based prospective screening of newborns differ from those observed in patients with clinical symptoms: identification and characterization of a new, prevalent mutation that resu. Am J Hum Genet. 2001;68:1408-18 pubmed
    ..A carrier frequency of 1/500 in the general population makes the 199T-->C mutation one of the three most prevalent mutations in the enzymes of fatty-acid oxidation. ..
  41. Parker A, Engel P. 5,5'-Dithiobis-(2-nitrobenzoic acid) as a probe for a non-essential cysteine residue at the medium chain acyl-coenzyme A dehydrogenase binding site of the human 'electron transferring flavoprotein' (ETF). J Enzyme Inhib. 1999;14:381-90 pubmed
    ..Conservation studies and the lack of 100% inactivation, however, suggested that this cysteine residue is not essential for the interaction with MCAD. ..
  42. Vreken P, van Lint A, Bootsma A, Overmars H, Wanders R, Van Gennip A. Rapid diagnosis of organic acidemias and fatty-acid oxidation defects by quantitative electrospray tandem-MS acyl-carnitine analysis in plasma. Adv Exp Med Biol. 1999;466:327-37 pubmed
  43. DuPlessis E, Pellett J, Stankovich M, Thorpe C. Oxidase activity of the acyl-CoA dehydrogenases. Biochemistry. 1998;37:10469-77 pubmed
    ..Evolutionary and mechanistic aspects of the suppression of oxygen reactivity in the acyl-CoA dehydrogenases are discussed. ..
  44. Corydon M, Vockley J, Rinaldo P, Rhead W, Kjeldsen M, Winter V, et al. Role of common gene variations in the molecular pathogenesis of short-chain acyl-CoA dehydrogenase deficiency. Pediatr Res. 2001;49:18-23 pubmed
    ..Our findings document that patients carrying these SCAD gene variations may develop clinically relevant SCAD deficiency, and that patients with even mild ethylmalonic aciduria should be tested for these variations. ..
  45. Fowler D, Picker J, Waisbren S, Levy H. Neonatal screening for medium--chain acyl-CoA dehydrogenase deficiency. Lancet. 2002;359:628 pubmed
  46. Van Hove J, Grunewald S, Jaeken J, Demaerel P, Declercq P, Bourdoux P, et al. D,L-3-hydroxybutyrate treatment of multiple acyl-CoA dehydrogenase deficiency (MADD). Lancet. 2003;361:1433-5 pubmed
    ..D,L-3-hydroxybutyrate is a therapeutic option for cerebral and cardiac complications in severe fatty acid oxidation defects. ..
  47. Han van der Kolk J, Gross J, Gerber V, Bruckmaier R. Disturbed bovine mitochondrial lipid metabolism: a review. Vet Q. 2017;37:262-273 pubmed publisher
    ..Assuming that liver carnitine concentrations might limit hepatic fatty acid oxidation capacity in dairy cows, further study of the role of acyl-CoA dehydrogenases and/or riboflavin in bovine ketosis is warranted...
  48. Yokota I, Coates P, Hale D, Rinaldo P, Tanaka K. Molecular survey of a prevalent mutation, 985A-to-G transition, and identification of five infrequent mutations in the medium-chain Acyl-CoA dehydrogenase (MCAD) gene in 55 patients with MCAD deficiency. Am J Hum Genet. 1991;49:1280-91 pubmed
    ..Of 29 patients whose country of origin was specified, 19 and five were from the British Isles and Germany, respectively. These data suggest that 985A----G may have occurred in a single person in an ancient Germanic tribe. ..
  49. Parker A, Engel P. Preliminary evidence for the existence of specific functional assemblies between enzymes of the beta-oxidation pathway and the respiratory chain. Biochem J. 2000;345 Pt 3:429-35 pubmed
    ..Pyridine haemochromogens, prepared from the partially purified species, show the presence of cytochrome proteins. The possible composition of these complexes and their relationship to the electron transport chain are discussed. ..
  50. McMahon B, Mayhew S. Identification and properties of an inducible phenylacyl-CoA dehydrogenase in Pseudomonas putida KT2440. FEMS Microbiol Lett. 2007;273:50-7 pubmed
    ..The experiments identify for the first time an acyl-CoA dehydrogenase that oxidizes the activated forms of aromatic acids similar to those used to first demonstrate the biological beta-oxidation of fatty acids. ..
  51. Hasegawa Y, Yamaguchi S. [Very-long-chain acyl-CoA dehydrogenase deficiency]. Nihon Rinsho. 2002;60 Suppl 4:717-21 pubmed
  52. Klose D, Kölker S, Heinrich B, Prietsch V, Mayatepek E, Von Kries R, et al. Incidence and short-term outcome of children with symptomatic presentation of organic acid and fatty acid oxidation disorders in Germany. Pediatrics. 2002;110:1204-11 pubmed
  53. Naito E, Indo Y, Tanaka K. Identification of two variant short chain acyl-coenzyme A dehydrogenase alleles, each containing a different point mutation in a patient with short chain acyl-coenzyme A dehydrogenase deficiency. J Clin Invest. 1990;85:1575-82 pubmed
    ..These C----T transitions result in the substitution of Arg-22 and Arg-83 of the mature SCAD with Trp and Cys, respectively. ..