ライフサイエンスコーパス: de novo

1 us missense mutations in WFS1 (4/5 confirmed de novo).

2 m and progenitor cells (HSPCs) are generated de novo.

3 canonical conformations and must be modeled de novo.

4 we demonstrate that parents transmitting the de novo 3 Mb LCR22A-D 22q11.2 deletion, the reciprocal d

5 Thus, we designed and tested simple de novo 98-residue polypeptides containing 7-residue rep

6 r 95 MHC homozygous cell lines we assembled, de novo, a set of high-fidelity contigs and a sequence s

7 atients with PsAF, 105 patients referred for de novo ablation of PsAF were prospectively recruited.

8 ites inactive in progenitors and reflect the de novo acquisition of AML-specific enhancers.

9 ase I cleavage, permitting identification of de novo active cis-regulatory modules (CRMs) and individ

10 Beige adipocytes can form through de novo adipogenesis; however, how "beiging" characteris

11 are formed that can act as seeds nucleating de novo aggregation of p53C.

12 53 mutations occur in less than one tenth of de novo AML cases.

13 de genotyping in 446 pediatric patients with de novo AML enrolled in Children's Oncology Group (COG)

14 n why the CSF3R mutations, which are rare in de novo AML, are so prevalent in SCN/AML.

15 SMAD6 had 4 de novo and 14 transmitted mutations; no other gene had

16 ation of 1q21 occurs in approximately 30% of de novo and 70% of relapsed multiple myeloma (MM) and is

17 De novo and acquired resistance, which are largely attri

18 tic mosaicism in healthy human tissues, link de novo and cancer mutations to somatic mosaicism, and c

19 mportant in determining helical stability in de novo and naturally-occurring polypeptides, giving new

20 ia (ALL) cells reveals substantial remaining de novo and salvage activities, and could not eliminate

21 rg helps explain the increased propensity of de novo Arg-rich SAHs to aggregate.

22 heoretically possible to obtain high-quality de novo assembled genome sequences but in practice DNA e

23 imately 400 million reads were generated and de novo assembled into 107,744 unigenes, with a mean len

24 We use the decision tree to classify de novo assembled sequences and compare the method to pu

25 We sequenced and de novo assembled the genomes of four cormorant species

26 32 transcripts with lengths over 200 bp were de novo assembled.

27 e reads produced by deep sequencing, such as de-novo assemblers, ignore the underlying diversity.

28 Some de novo assemblies have been constructed free of referen

29 We report de novo assemblies of 150 individuals (50 trios) from th

30 ods when classifying sequences in eukaryotic de novo assemblies.

31 Further improvement of de novo assembly algorithms are needed in order to warra

32 Recent progress in de novo assembly and whole-genome resequencing of wild a

33 We determine the optimal de novo assembly methods and parameter combinations requ

34 are) capable of accurate, massively parallel de novo assembly of high quality DNA barcodes of >1400 b

35 MECAT enables reference mapping or de novo assembly of large genomes using SMS reads on a s

36 De novo assembly of whole genome shotgun (WGS) next-gene

37 De novo assembly produced 67,911 transcripts with a high

38 text searchable index for read alignment and de novo assembly.

39 Here we present a de novo atomic structure of Drosophila NOMPC determined

40 Here, we show that the majority of primary (de novo) atypical meningiomas display loss of NF2, which

41 Recently, we characterized a de novo autism-linked UBE3A mutant (UBE3A(T485A)) that d

42 His theories, however, did not arise de novo, being strongly influenced by Karl Kahlbaum and

43 e translation reporter analysis reveals that de novo beta-actin hotspots colocalize with docked RNA g

44 d formation of ILVs, we exploited the rapid, de novo biogenesis of MVEs during the oocyte-to-embryo t

45 Cells acquire sphingolipids by both de novo biosynthesis and recycling of exogenous sphingol

46 hat phloem and xylem might contribute to the de novo biosynthesis of JA after fungal infection.

47 ER stress leads to de novo biosynthesis of non-trimethylated GRP78, whereas

48 hase (TS) is the sole enzyme responsible for de novo biosynthesis of thymidylate (TMP) and is essenti

49 have demonstrated the presence of an intact de novo biosynthesis pathway for NAD(+) from tryptophan

50 Additionally, a de novo c.1066G>A (p.Ala356Thr) variant was identified i

51 %) from recurrent cancer and 27 (10.5%) from de novo cancer.

52 In vivo ceramide reduction by inhibition of de novo ceramide synthesis reduced PLIN2 and hepatic ste

53 ; ovarian/endometrial/vulvar cancers, 3; and de novo cholangiocarcinoma, 4).

54 ed controls and no difference in the rate of de novo CNV development in eyes with or without VMA.

55 contrast, there is no general method for the de novo computational design of multicross-linked protei

56 igestion with three proteases and sequencing de novo defined the complete protein sequence and locate

57 sis: orienting dendrites toward TCAs, adding de novo dendritic segments, and elongating dendritic len

58 lpha/beta and alpha+beta protein classes via de novo design and cooperative assembly strategies.

59 De novo design of antibodies binding specific epitopes c

60 have profound impact on the optimization and de novo design of new NM with better in vivo performance

61 ffect of a GXXXG motif on the association of de novo designed (AALALAA)3 helices in liposomes.

62 from oligodeoxyribonucleotides a completely de-novo-designed, 58-kb multigene DNA.

63 De novo digenic mutations of telomere-associated protein

64 cal imaging limitations makes the systematic de novo discovery of structures within cells challenging

65 To date, most de novo discovery through next-generation sequencing foc

66 period length through a mechanism requiring de novo DNA methylation.

67 ding DNA methyltransferases where it imposes de novo DNA methylation.

68 recruiting multiple copies of antibody-fused de novo DNA methyltransferase 3A (DNMT3A) (dCas9-SunTag-

69 (mESCs), p53 restricts the expression of the de novo DNA methyltransferases Dnmt3a and Dnmt3b while u

70 re, we present an economical and streamlined de novo DNA synthesis approach for engineering a synthet

71 These data establish de novo DNA-methylation programming as a regulator of T

72 There have been several reports linking de novo dominant EEF1A2 mutations to neurological issues

73 the development of interstitial fibrosis and de novo donor specific anti-HLA antibodies (dnDSA) at 1

74 analyzed were: HLA antibodies at transplant, de novo donor-specific antibodies (DSA), antibody-mediat

75 We hypothesized that HLA class II de novo donor-specific antibody (dnDSA) development corr

76 We hypothesized that de novo donor-specific antibody (DSA) causes complement-

77 ad superior graft survival compared with the de novo DSA ABMR (63% versus 34% at 8 years after reject

78 De novo DSA ABMR was characterized by increased expressi

79 DQ mismatch is a significant risk factor for de novo DSA emergence, whereas the persistence of antibo

80 s of early graft failure, whereas those with de novo DSA experienced accelerated graft loss once DSA

81 An additional 24 (25%) patients developed de novo DSA, and of these, 71% had persistent antibodies

82 antibodies (DSA) or in patients who develop de novo DSA.

83 s in vitamin B12 depletion, which suppresses de novo dTMP biosynthesis and causes DNA damage, account

84 generate 5,10-methylenetetrahydrofolate for de novo dTMP biosynthesis and translocate to the nucleus

85 Vitamin B12 depletion decreased de novo dTMP biosynthesis capacity by 5-35%, whereas de

86 Furthermore, we show that de novo dTMP synthesis does not occur in the cytosol at

87 This computational model shows that de novo dTMP synthesis is highly sensitive to the common

88 irical data indicating that impaired nuclear de novo dTMP synthesis results in uracil misincorporatio

89 Here, using de novo dual-RNA seq, we compared the host salamander ce

90 +) produced oligodendrocytes responsible for de novo ensheathment of approximately 30% of myelinated

91 cells reconstitute adult articular cartilage de novo, entirely substituting fetal chondrocytes.

92 Instead de novo enzymatic reactions could more productively bene

93 ultimately a limited strategy for improving de novo enzymes since it is largely restricted to optimi

94 De novo expression in the kidney of periostin, a protein

95 nsgenic mouse models for tamoxifen-inducible de novo expression of Ags in LCs but no other langerin(+

96 Here we found that de novo expression of Peg3 increased Beclin 1 promoter a

97 o sequence and assemble new species' genomes de novo fail to achieve the ultimate endpoint to produce

98 nstrate that BPL-1 is required for efficient de novo fatty acid biosynthesis.

99 iencies rather than fuel types, reflecting a de novo formation mechanism.

100 ons intersecting with a TAD boundary mediate de novo formation of a 3D contact domain comprising IGF2

101 ing bacteria, in which new offspring emerges de novo from a morphologically invariant mother cell.

102 DNA damage response, and implicated Vts1 in de novo gene "birth." TGA provided single-nucleotide res

103 ch Initiative, and Open Targets, cover human de novo gene variants, disease-related phenotypes in mod

104 t the gastrointestinal tract is deficient in de novo generation of Treg cells in allergic mice.

105 Here, we present draft de novo genome and plastome assemblies for a wilt-resist

106 roup had a lower failure rate, lower rate of de novo glaucoma surgery, and lower mean IOP on fewer me

107 Here we show that a de novo Glu183 to Val (E183V) mutation in the CaMKIIalph

108 We found that TCF19 overexpression represses de novo glucose production in HepG2 cells.

109 d with a lower response rate compared to the de novo group (57% versus 69%, P univariate=0.008, P mul

110 present a challenge to SIR complex-mediated de novo heterochromatic silencing due to the presence of

111 These mutations included five de novo heterozygous loss of function mutations/deletion

112 plantation (LT) recipients with preformed or de novo HLA donor-specific alloantibodies (DSA).

113 r cells, both murine and human CAFs promoted de novo HT resistance via the generation of CD133(hi) CS

114 Enamel crystals form de novo in a rich extracellular environment in a stage-d

115 encoding p.Glu210Lys in UBF, which occurred de novo in all cases.

116 s in EBF3 were found; the mutations occurred de novo in eight individuals, and the missense variant c

117 In cells, Coenzyme A is synthesized de novo in five enzymatic steps with vitamin B5 as the s

118 Because language can be invented de novo in the manual modality, this offers insight into

119 The mutation was confirmed as de novo in three of the cases, and the mildly affected f

120 Our current work shows that de novo induction is more complex than previously though

121 This de novo induction of [PSI (+)] shows the appearance of f

122 IL-1beta depends on 2 regulated events: the de novo induction of pro-IL-1beta, generally via NF-kapp

123 opulation structure and how self-renewal and de novo influx contribute to the maintenance of memory c

124 de novo single nucleotide variants and 12.6 de novo insertions and deletions or copy number variatio

125 n of neurons, but these mice did not develop de novo insoluble tau aggregates, which are characterist

126 These were assembled de novo into 138,183 unique contigs.

127 We reasoned that de novo introduction of CD22 ligands in RBCs should abol

128 De novo KCNB1 missense variants in the ion channel domai

129 estimate based on LDE applied to comparable de novo Kemp eliminases and other enzymes like KSI.

130 ly promoting LapA proteolysis and preventing de novo LapA synthesis and secretion.

131 We also construct de novo linkage maps on 7-12x whole-genome data on the R

132 r pharmacological inhibition of SRPK2 blunts de novo lipid synthesis, thereby suppressing cell growth

133 Hepatic de novo lipogenesis (DNL) converts carbohydrates into tr

134 eases serum glucose levels, which stimulates de novo lipogenesis (DNL) in the liver.

135 s liver lipid metabolism by inhibiting liver de novo lipogenesis as a downstream player of the p63 ne

136 The metabolic pathway of de novo lipogenesis is frequently upregulated in human l

137 could be explained in part by an increase in de novo lipogenesis that results from increased sterol e

138 eveloped steatosis upon induction of hepatic de novo lipogenesis with fructose feeding.

139 ene CIDEA as well as other genes involved in de novo lipogenesis.

140 logical down-regulation to prevent excessive de novo lipogenesis.

141 ver, we successfully apply o2n-seq to screen de novo, low-frequency mutations in human tumours.

142 quiring organisms are able to synthesize B12 de novo, making it a common commodity in microbial commu

143 cancer-specific mortality and of developing de novo malignancies after transplantation compared with

144 in survival and incidence of posttransplant de novo malignancies exist between recipients with PTM a

145 and maintenance methylated MB corresponds to de novo methylation event.

146 on to memory cells was coupled to erasure of de novo methylation programs and re-expression of naive-

147 identified progressively acquired heritable de novo methylation programs that restrict T cell expans

148 mechanisms by which CGIs are protected from de novo methylation remain elusive.

149 Only a few cases of de novo missense mutations in EHMT1 giving rise to KS ha

150 We identified the properties of de novo missense mutations in patients with neurodevelop

151 Here de novo missense mutations in the RPS23 gene, which code

152 We report seven individuals with distinct de novo missense RAC1 mutations and varying degrees of d

153 independent patients, each of whom carried a de novo missense variant in srp54 (encoding signal recog

154 We have now identified de novo missense variants clustering in the BTB-domain-e

155 De novo missense variants explained a larger proportion

156 dently derived logistic regression models (a de novo model and an a priori model developed using elec

157 adult mammals, large wounds in mice lead to de novo morphogenesis of hair follicles.

158 e predicted the enrichment of both known and de novo motifs based on ChIP data.

159 Analysis of nonhuman organisms enabled de novo mtDNA sequence assembly, as well as detection of

160 in two dominant FEVR-affected families and a de novo mutation (c.1434_1435insC [p.Glu479Argfs( *)18])

161 th severe encephalopathy carrying a missense de novo mutation in GRIN2B(p.P553T) coding for the GluN2

162 Vs, we explored the functional impact of the de novo mutation in TBL1XR1 [c.30 C > G (p.Phe10Leu)], a

163 henomenon relates to genome-wide patterns of de novo mutation.

164 understand the impact of disease-associated de novo mutations and other rare sequence variants on TR

165 We identified 7.5% of de novo mutations as PZMs, 83.3% of which were not descr

166 re we show that the same mutations as inborn de novo mutations cause an early onset multisystem disor

167 wide association studies, approximately 1000 de novo mutations discovered by large-scale sequencing o

168 Previously identified putative de novo mutations failed to complement yeast strains lac

169 by a novel mathematical model incorporating de novo mutations for both species.

170 nerate a data set for this purpose using (1) de novo mutations from mutation accumulation experiments

171 Overall, we find significant clustering of de novo mutations in 200 genes, highlighting specific fu

172 Here, we report that de novo mutations in EBF3 cause a complex neurodevelopme

173 etic aetiology and are often associated with de novo mutations in genes mediating synaptic transmissi

174 h significant spatial clustering patterns of de novo mutations in large cohorts.

175 s where genetic causes have been identified, de novo mutations in neuronally expressed genes are a co

176 enriched for genes that harbor nonsynonymous de novo mutations in patients with epileptic encephalopa

177 in the global influenza population begin as de novo mutations in single infected hosts, but the evol

178 geted search, we identified an enrichment of de novo mutations in the gene encoding the 330-kDa tripl

179 Recently, de novo mutations in the gene KCNA2, causing either a do

180 , we discover a large cluster of ASD-related de novo mutations in Trio's Rac1 activating domain, GEF1

181 Children with ASD with de novo mutations may exhibit a "muted" symptom profile

182 sm indicating that a large fraction of these de novo mutations occurred during early germ cell develo

183 s describing the genetic association between de novo mutations of NMDAR subunits and severe psychiatr

184 Resistance was associated with de novo mutations, rather than acquisition of resistant

185 le, we sequenced them in parents to identify de novo mutations.

186 5% of NSC is sporadic, suggesting a role for de novo mutations.

187 Eight patients had de novo mutations.

188 r cells, oligodendrocyte differentiation and de novo myelination of parallel fibers.

189 a powerful tool to evaluate the prognosis of de novo myeloma.

190 llae lack the nadA and nadB genes needed for de novo NAD biosynthesis, remarkably, they have one de n

191 absence of supplied pyridines, indicative of de novo NAD synthesis and functional confirmation of Bor

192 aenorhabditis elegans are reported to lack a de novo NAD(+) biosynthetic pathway.

193 ase undergoing planned stent implantation in de-novo, native coronary lesions.

194 Biopsies with de novo non-HLA autoantibodies revealed a new sinusoidal

195 Here we report de novo non-synonymous single-nucleotide variants (SNVs)

196 ), initially diagnosed with OTCS, revealed a de novo nonsense mutation, p.Q638*, in the MAGEL2 gene.

197 seven unrelated individuals with a recurrent de novo nonsense variant (c.2737C>T [p.Arg913Ter]) in th

198 ratio were associated with increased risk of de novo (nonskin) malignancy.

199 Recurrent de novo or case-private CNVs were found at 15q11-13, Xp2

200 The distinction between de novo or recently diagnosed HF and worsening chronic H

201 Moreover, de novo oscillating genes under CR show an enrichment in

202 ective cohort of, at baseline, patients with de novo Parkinson's disease (PD) compared with healthy c

203 ticentre international longitudinal study of de novo Parkinson's disease.

204 We report 15 individuals with de novo pathogenic variants in WDR26.

205 NAD biosynthesis, remarkably, they have one de novo pathway gene, nadC, encoding quinolinate phospho

206 Alternatively, de novo peptide sequencing algorithms annotate MS/MS spe

207 We describe a strategy for de novo peptide sequencing based on matched pairs of tan

208 nt of replicative repair, and show increased de novo point mutations flanking the rearrangement junct

209 Here, the authors show that de novo polarisation of the mouse embryo at the 8-cell s

210 Considering only top-ranked de novo predictions, 70% of the pairs were deciphered co

211 nctions, thus facilitating lateral spread of de novo-produced virions.

212 ient MEFs with type I IFN, illustrating that de novo production of IFN-beta in response to VSV plays

213 nderstanding protein functions inside cells, de novo protein design as well as defining the role of p

214 De novo protein design holds promise for creating small

215 rating a principle predicted from efforts in de novo protein design.

216 of STAT phosphorylation that requires SMADs, de novo protein synthesis, and contribution from JAK1.

217 consolidation of extinction memory requires de novo protein synthesis.

218 S)-based techniques to separate and sequence de novo proto-peptides containing broader combinations o

219 on of cvpA-purF, or of purF alone, abolished de novo purine biosynthesis but did not impact bacterial

220 the second enzyme in the guanylate branch of de novo purine biosynthesis.

221 within the intracellular niche and relies on de novo purine synthesis to meet this metabolic requirem

222 on in human BTICs to a nexus between MYC and de novo purine synthesis, mediating glucose-sustained an

223 dTMP biosynthesis capacity by 5-35%, whereas de novo purine synthesis, which occurs in the cytosol, w

224 lls rely on RC-derived aspartate to maintain de novo pyrimidine biosynthesis.

225 ty of this NS5 protein is verified through a de novo RdRp assay on a subgenomic ZIKV RNA template.

226 ariants were taken forward for in silico and de novo replication (11 common and 5 rare).

227 However, mCRPC showed overwhelming de novo resistance to immune checkpoint blockade, motiva

228 a subset of genes that adopted increased or de novo rhythmicity during aging, enriched for stress-re

229 ither ribosome recycling on the same mRNA or de novo ribosome recruitment.

230 provides an organ regeneration model through de novo root formation, stating key stages and the prima

231 Here, de novo Rosetta modeling and competitive binding experim

232 Clinical predictors for recurrent de novo SCAD were tested using univariate and multivaria

233 Similar functional deficits were found for a de novo SCZ variant GIT1-S601N.

234 eaf, stem, and flower tissues, and performed de novo sequence assembly, yielding 98,613 unique transc

235 However, Arg-rich de novo sequences (ER3 (AEEERRR) and EK1R2 (AEEEKRR)) ag

236 chnology to oligosaccharide standards and to de novo sequencing of purified plant metabolite glycocon

237 This remarkable property is applied to de novo sequencing of underivatized oligosaccharides.Est

238 This de novo sequencing performance exceeds that of PEAKS and

239 results illustrate the power of whole-genome de novo sequencing relative to resequencing and provide

240 these paired spectra, we modified the UVnovo de novo sequencing software to automatically learn from

241 experimental and computational approach for de novo sequencing using whole cell E. coli lysate.

242 While some cancer cells upregulate de novo serine synthesis, many others rely on exogenous

243 ted that minor resistant variants may emerge de novo shortly after transmission, when the small effec

244 NE_Scan outperforms the previously published de novo SINE discovery program.

245 We found an average of 73.8 de novo single nucleotide variants and 12.6 de novo inse

246 kely pathogenic status without knowing their de novo status.

247 still of low quality and not very useful for de novo structure prediction.

248 ma8, combining recent structural studies and de novo structure predictions.

249 symptomatic peripheral artery disease due to de novo superficial femoral artery stenotic or occlusive

250 rrects hyperactivity, seizures, and elevated de novo synaptic protein synthesis.

251 atic steatosis in alcohol-fed mice, but only de novo synthesis inhibition, not sphingomyelin hydrolys

252 c pathways, including those required for the de novo synthesis of dTMP and purine nucleotides and for

253 ompounds require accurate and cost-effective de novo synthesis of genetic pathways.

254 BACKGROUND & AIMS: De novo synthesis of guanosine diphosphate (GDP)-fucose,

255 The highly stereocontrolled de novo synthesis of l-NBDNJ (the unnatural enantiomer o

256 t on the level of phosphatidate used for the de novo synthesis of membrane phospholipids.

257 Solvent assisted linker exchange (SALE) and de novo synthesis of mixed-linker ZIFs have been demonst

258 to exogenously supplied fatty acids via the de novo synthesis of PA, a central metabolite for membra

259 s dependent upon the enzymes responsible for de novo synthesis of PA.

260 ept that glycerophosphoinositol inhibits the de novo synthesis of proinflammatory and prothrombotic c

261 sential vitamins and cofactors and decreased de novo synthesis of pyrimidines in cycle two.

262 NKPD1 is predicted to be involved in the de novo synthesis of sphingolipids, which have been impl

263 The ability to de novo synthesize purines has been associated with the

264 s found to be a continuous 8:2 ratio between de novo synthesized FA and acyl chain transfer from pre-

265 Our design comprised de novo synthetic cancer-specific promoters and, to enha

266 ly, including biomimetic, semisynthetic, and de novo synthetic strategies.

267 Conversely, de novo T3 that can be formed upon iodination of TG secr

268 These results suggest that a de novo TBL1XR1 point mutation could alter Wnt/beta-cate

269 puter-derived nucleotide sequence is evolved de novo that is optimal for packaging the RNA into capsi

270 odifier (SUMO)- and folate-dependent nuclear de novo thymidylate (dTMP) biosynthesis is a sensitive t

271 e (5-methylTHF) and subsequent inhibition of de novo thymidylate (dTMP) biosynthesis.

272 l tube defects (NTDs) indicate that impaired de novo thymidylate (dTMP) synthesis through changes in

273 mma, and of IL-2, depends on calcium-induced de novo transcription and PKC-dependent mRNA stabilizati

274 We measured the first wave of de novo transcription in living Drosophila embryos using

275 De novo transcriptome assembly is an important technique

276 In this study, we combine de novo transcriptome data and sequenced genomes from an

277 hput sequencing (HTS), we obtained the first de novo transcriptome of mahi-mahi, with 60,842 assemble

278 Through de novo transcriptome reconstruction, we establish dynam

279 r spikemoss Selaginella moellendorffii Using de novo transcriptomics, we confirmed expression of five

280 nd that low to mild doses of oxidants induce de novo translation of the NRF2 protein.

281 to severe ID and/or developmental delay and de novo truncating PPM1D mutations.

282 De novo use of mammalian target of rapamycin inhibitors

283 this chromatin architecture can be predicted de novo using epigenetic data derived from chromatin imm

284 approach for predicting peptidase substrates de novo using protein structure modeling and biophysical

285 isk in ASD cases who carry a strongly acting de novo variant.

286 for a better understanding of the biology of de novo variants and their contribution to disease.

287 rtance of investigating the pathogenicity of de novo variants because they are not as exclusive and p

288 db contained 40 different studies and 32,991 de novo variants from 23,098 trios.

289 mental disorders, we suggest that pathogenic de novo variants in GRIA4 lead to intellectual disabilit

290 ave facilitated the large-scale discovery of de novo variants in human disease.

291 De novo variants were found only in SLC1A2 (aka EAAT2 or

292 , we created denovo-db, a database for human de novo variants.

293 research has uncovered an important role for de novo variation in neurodevelopmental disorders.

294 The importance of de novo variation in sporadic disease is a recent highli

295 ) risk is influenced by common polygenic and de novo variation.

296 response and survival in patients receiving de novo versus upgrade CRT defibrillator therapy.

297 nd without antiviral therapy, which prevents de novo viral replication.

298 of HCV dsRNA through a process that required de novo viral RNA synthesis and shifted the ratio of vir

299 All mutations were de novo where parental DNA was available.

300 ed with BAC-end sequences (BES) to produce a de novo whole-genome physical map.