Details of Drug Off-Target (DOT)

General Information of Drug Off-Target (DOT) (ID: OTH5BPLO)

• DOT Name: Partitioning defective 3 homolog (PARD3)
• Synonyms: PAR-3; PARD-3; Atypical PKC isotype-specific-interacting protein; ASIP; CTCL tumor antigen se2-5; PAR3-alpha
• Gene Name: PARD3
• Related Disease:
  Advanced cancer
  Alzheimer disease
  Breast cancer
  Clear cell renal carcinoma
  Coeliac disease
  Colorectal adenocarcinoma
  Colorectal carcinoma
  Endometrial cancer
  Endometrial carcinoma
  Epithelial ovarian cancer
  Glioblastoma multiforme
  Invasive breast carcinoma
  Neoplasm
  Neural tube defect
  Ovarian cancer
  Ovarian neoplasm
  Pulmonary fibrosis
  Renal cell carcinoma
  Rhabdomyosarcoma
  Skin neoplasm
  Squamous cell carcinoma
  Thyroid gland undifferentiated (anaplastic) carcinoma
  Triple negative breast cancer
  Ulcerative colitis
  Zika virus infection
  Gastric cancer
  Graft-versus-host disease
  Metastatic malignant neoplasm
  Obesity
  Prostate carcinoma
  Stomach cancer
  Arthritis
  Cutaneous squamous cell carcinoma
  Melanoma
  Neural tube defects, susceptibility to
  Prostate cancer
  Prostate neoplasm
• UniProt ID: PARD3_HUMAN
• PDB ID: 2KOM
• Pfam ID: PF12053 ; PF00595
• Sequence:
  MKVTVCFGRTRVVVPCGDGHMKVFSLIQQAVTRYRKAIAKDPNYWIQVHRLEHGDGGILD
  LDDILCDVADDKDRLVAVFDEQDPHHGGDGTSASSTGTQSPEIFGSELGTNNVSAFQPYQ
  ATSEIEVTPSVLRANMPLHVRRSSDPALIGLSTSVSDSNFSSEEPSRKNPTRWSTTAGFL
  KQNTAGSPKTCDRKKDENYRSLPRDTSNWSNQFQRDNARSSLSASHPMVGKWLEKQEQDE
  DGTEEDNSRVEPVGHADTGLEHIPNFSLDDMVKLVEVPNDGGPLGIHVVPFSARGGRTLG
  LLVKRLEKGGKAEHENLFRENDCIVRINDGDLRNRRFEQAQHMFRQAMRTPIIWFHVVPA
  ANKEQYEQLSQSEKNNYYSSRFSPDSQYIDNRSVNSAGLHTVQRAPRLNHPPEQIDSHSR
  LPHSAHPSGKPPSAPASAPQNVFSTTVSSGYNTKKIGKRLNIQLKKGTEGLGFSITSRDV
  TIGGSAPIYVKNILPRGAAIQDGRLKAGDRLIEVNGVDLVGKSQEEVVSLLRSTKMEGTV
  SLLVFRQEDAFHPRELNAEPSQMQIPKETKAEDEDIVLTPDGTREFLTFEVPLNDSGSAG
  LGVSVKGNRSKENHADLGIFVKSIINGGAASKDGRLRVNDQLIAVNGESLLGKTNQDAME
  TLRRSMSTEGNKRGMIQLIVARRISKCNELKSPGSPPGPELPIETALDDRERRISHSLYS
  GIEGLDESPSRNAALSRIMGESGKYQLSPTVNMPQDDTVIIEDDRLPVLPPHLSDQSSSS
  SHDDVGFVTADAGTWAKAAISDSADCSLSPDVDPVLAFQREGFGRQSMSEKRTKQFSDAS
  QLDFVKTRKSKSMDLGIADETKLNTVDDQKAGSPSRDVGPSLGLKKSSSLESLQTAVAEV
  TLNGDIPFHRPRPRIIRGRGCNESFRAAIDKSYDKPAVDDDDEGMETLEEDTEESSRSGR
  ESVSTASDQPSHSLERQMNGNQEKGDKTDRKKDKTGKEKKKDRDKEKDKMKAKKGMLKGL
  GDMFRFGKHRKDDKIEKTGKIKIQESFTSEEERIRMKQEQERIQAKTREFRERQARERDY
  AEIQDFHRTFGCDDELMYGGVSSYEGSMALNARPQSPREGHMMDALYAQVKKPRNSKPSP
  VDSNRSTPSNHDRIQRLRQEFQQAKQDEDVEDRRRTYSFEQPWPNARPATQSGRHSVSVE
  VQMQRQRQEERESSQQAQRQYSSLPRQSRKNASSVSQDSWEQNYSPGEGFQSAKENPRYS
  SYQGSRNGYLGGHGFNARVMLETQELLRQEQRRKEQQMKKQPPSEGPSNYDSYKKVQDPS
  YAPPKGPFRQDVPPSPSQVARLNRLQTPEKGRPFYS
• Function: Adapter protein involved in asymmetrical cell division and cell polarization processes. Seems to play a central role in the formation of epithelial tight junctions. Targets the phosphatase PTEN to cell junctions. Involved in Schwann cell peripheral myelination. Association with PARD6B may prevent the interaction of PARD3 with F11R/JAM1, thereby preventing tight junction assembly. The PARD6-PARD3 complex links GTP-bound Rho small GTPases to atypical protein kinase C proteins. Required for establishment of neuronal polarity and normal axon formation in cultured hippocampal neurons.
• Tissue Specificity: Widely expressed.
• KEGG Pathway:
  Rap1 sig.ling pathway (hsa04015)
  Chemokine sig.ling pathway (hsa04062)
  Endocytosis (hsa04144)
  Axon guidance (hsa04360)
  Hippo sig.ling pathway (hsa04390)
  Adherens junction (hsa04520)
  Tight junction (hsa04530)
  Human papillomavirus infection (hsa05165)
• Reactome Pathway:
  Tight junction interactions (R-HSA-420029)
  TGF-beta receptor signaling in EMT (epithelial to mesenchymal transition) (R-HSA-2173791)

Molecular Interaction Atlas (MIA) of This DOT

37 Disease(s) Related to This DOT

Disease Name	Disease ID	Evidence Level	Mode of Inheritance	REF
Advanced cancer	DISAT1Z9	Strong	Biomarker	[1]
Alzheimer disease	DISF8S70	Strong	Biomarker	[2]
Breast cancer	DIS7DPX1	Strong	Altered Expression	[3]
Clear cell renal carcinoma	DISBXRFJ	Strong	Altered Expression	[4]
Coeliac disease	DISIY60C	Strong	Biomarker	[5]
Colorectal adenocarcinoma	DISPQOUB	Strong	Altered Expression	[6]
Colorectal carcinoma	DIS5PYL0	Strong	Biomarker	[7]
Endometrial cancer	DISW0LMR	Strong	Altered Expression	[8]
Endometrial carcinoma	DISXR5CY	Strong	Altered Expression	[8]
Epithelial ovarian cancer	DIS56MH2	Strong	Biomarker	[9]
Glioblastoma multiforme	DISK8246	Strong	Biomarker	[10]
Invasive breast carcinoma	DISANYTW	Strong	Biomarker	[3]
Neoplasm	DISZKGEW	Strong	Biomarker	[11]
Neural tube defect	DIS5J95E	Strong	Biomarker	[12]
Ovarian cancer	DISZJHAP	Strong	Altered Expression	[13]
Ovarian neoplasm	DISEAFTY	Strong	Altered Expression	[13]
Pulmonary fibrosis	DISQKVLA	Strong	Biomarker	[14]
Renal cell carcinoma	DISQZ2X8	Strong	Biomarker	[15]
Rhabdomyosarcoma	DISNR7MS	Strong	Biomarker	[16]
Skin neoplasm	DIS16DDV	Strong	Biomarker	[1]
Squamous cell carcinoma	DISQVIFL	Strong	Altered Expression	[17]
Thyroid gland undifferentiated (anaplastic) carcinoma	DISYBB1W	Strong	Biomarker	[18]
Triple negative breast cancer	DISAMG6N	Strong	Biomarker	[19]
Ulcerative colitis	DIS8K27O	Strong	Genetic Variation	[20]
Zika virus infection	DISQUCTY	Strong	Altered Expression	[21]
Gastric cancer	DISXGOUK	moderate	Altered Expression	[22]
Graft-versus-host disease	DIS0QADF	moderate	Biomarker	[23]
Metastatic malignant neoplasm	DIS86UK6	moderate	Biomarker	[24]
Obesity	DIS47Y1K	moderate	Biomarker	[25]
Prostate carcinoma	DISMJPLE	moderate	Biomarker	[24]
Stomach cancer	DISKIJSX	moderate	Altered Expression	[22]
Arthritis	DIST1YEL	Disputed	Altered Expression	[26]
Cutaneous squamous cell carcinoma	DIS3LXUG	Limited	Biomarker	[27]
Melanoma	DIS1RRCY	Limited	Biomarker	[28]
Neural tube defects, susceptibility to	DISHA84K	Limited	SusceptibilityMutation	[29]
Prostate cancer	DISF190Y	Limited	Biomarker	[30]
Prostate neoplasm	DISHDKGQ	Limited	Genetic Variation	[31]

This DOT Affected the Drug Response of 1 Drug(s)

Drug Name	Drug ID	Highest Status	Interaction	REF
Fluorouracil	DMUM7HZ	Approved	Partitioning defective 3 homolog (PARD3) affects the response to substance of Fluorouracil.	[49]

6 Drug(s) Affected the Post-Translational Modifications of This DOT

Drug Name	Drug ID	Highest Status	Interaction	REF
Valproate	DMCFE9I	Approved	Valproate increases the methylation of Partitioning defective 3 homolog (PARD3).	[32]
Arsenic	DMTL2Y1	Approved	Arsenic affects the methylation of Partitioning defective 3 homolog (PARD3).	[39]
Quercetin	DM3NC4M	Approved	Quercetin decreases the phosphorylation of Partitioning defective 3 homolog (PARD3).	[40]
PMID28870136-Compound-52	DMFDERP	Patented	PMID28870136-Compound-52 affects the phosphorylation of Partitioning defective 3 homolog (PARD3).	[40]
Bisphenol A	DM2ZLD7	Investigative	Bisphenol A increases the methylation of Partitioning defective 3 homolog (PARD3).	[47]
Coumarin	DM0N8ZM	Investigative	Coumarin affects the phosphorylation of Partitioning defective 3 homolog (PARD3).	[40]

16 Drug(s) Affected the Gene/Protein Processing of This DOT

Drug Name	Drug ID	Highest Status	Interaction	REF
Ciclosporin	DMAZJFX	Approved	Ciclosporin increases the expression of Partitioning defective 3 homolog (PARD3).	[33]
Tretinoin	DM49DUI	Approved	Tretinoin decreases the expression of Partitioning defective 3 homolog (PARD3).	[34]
Doxorubicin	DMVP5YE	Approved	Doxorubicin decreases the expression of Partitioning defective 3 homolog (PARD3).	[35]
Cupric Sulfate	DMP0NFQ	Approved	Cupric Sulfate decreases the expression of Partitioning defective 3 homolog (PARD3).	[36]
Cisplatin	DMRHGI9	Approved	Cisplatin decreases the expression of Partitioning defective 3 homolog (PARD3).	[37]
Ivermectin	DMDBX5F	Approved	Ivermectin decreases the expression of Partitioning defective 3 homolog (PARD3).	[38]
Testosterone enanthate	DMB6871	Approved	Testosterone enanthate affects the expression of Partitioning defective 3 homolog (PARD3).	[41]
Amphotericin B	DMTAJQE	Approved	Amphotericin B decreases the expression of Partitioning defective 3 homolog (PARD3).	[37]
Permethrin	DMZ0Q1G	Approved	Permethrin increases the expression of Partitioning defective 3 homolog (PARD3).	[42]
Cyclophosphamide	DM4O2Z7	Approved	Cyclophosphamide decreases the expression of Partitioning defective 3 homolog (PARD3).	[37]
Gentamicin	DMKINJO	Approved	Gentamicin decreases the expression of Partitioning defective 3 homolog (PARD3).	[37]
Tocopherol	DMBIJZ6	Phase 2	Tocopherol decreases the expression of Partitioning defective 3 homolog (PARD3).	[43]
Benzo(a)pyrene	DMN7J43	Phase 1	Benzo(a)pyrene decreases the expression of Partitioning defective 3 homolog (PARD3).	[44]
Leflunomide	DMR8ONJ	Phase 1 Trial	Leflunomide increases the expression of Partitioning defective 3 homolog (PARD3).	[45]
PMID28460551-Compound-2	DM4DOUB	Patented	PMID28460551-Compound-2 increases the expression of Partitioning defective 3 homolog (PARD3).	[46]
Coumestrol	DM40TBU	Investigative	Coumestrol decreases the expression of Partitioning defective 3 homolog (PARD3).	[48]

References

• [1] Shared and independent functions of aPKC and Par3 in skin tumorigenesis.Oncogene. 2018 Sep;37(37):5136-5146. doi: 10.1038/s41388-018-0313-1. Epub 2018 May 23.
• [2] Par3 and aPKC regulate BACE1 endosome-to-TGN trafficking throughPACS1.Neurobiol Aging. 2017 Dec;60:129-140. doi: 10.1016/j.neurobiolaging.2017.08.024. Epub 2017 Sep 21.
• [3] Loss of the Par3 polarity protein promotes breast tumorigenesis and metastasis.Cancer Cell. 2012 Nov 13;22(5):601-14. doi: 10.1016/j.ccr.2012.10.003.
• [4] Independent association of PD-L1 expression with noninactivated VHL clear cell renal cell carcinoma-A finding with therapeutic potential.Int J Cancer. 2017 Jan 1;140(1):142-148. doi: 10.1002/ijc.30429. Epub 2016 Sep 23.
• [5] Intestinal barrier gene variants may not explain the increased levels of antigliadin antibodies, suggesting other mechanisms than altered permeability.Hum Immunol. 2010 Apr;71(4):392-6. doi: 10.1016/j.humimm.2010.01.016. Epub 2010 Feb 4.
• [6] Decreased Expression of the Polarity Regulatory PAR Complex Predicts Poor Prognosis of the Patients with Colorectal Adenocarcinoma.Transl Oncol. 2018 Feb;11(1):109-115. doi: 10.1016/j.tranon.2017.11.004. Epub 2017 Dec 18.
• [7] LncRNA SLCO4A1-AS1 promotes colorectal cancer cell proliferation by enhancing autophagy via miR-508-3p/PARD3 axis.Aging (Albany NY). 2019 Jul 16;11(14):4876-4889. doi: 10.18632/aging.102081.
• [8] Loss of polarity alters proliferation and differentiation in low-grade endometrial cancers by disrupting Notch signaling.PLoS One. 2017 Dec 5;12(12):e0189081. doi: 10.1371/journal.pone.0189081. eCollection 2017.
• [9] Tissue factor-factor VIIa complex triggers protease activated receptor 2-dependent growth factor release and migration in ovarian cancer.Gynecol Oncol. 2017 Apr;145(1):167-175. doi: 10.1016/j.ygyno.2017.01.022. Epub 2017 Jan 29.
• [10] A genome-wide screen for microdeletions reveals disruption of polarity complex genes in diverse human cancers.Cancer Res. 2010 Mar 15;70(6):2158-64. doi: 10.1158/0008-5472.CAN-09-3458. Epub 2010 Mar 9.
• [11] Pard3 suppresses glioma invasion by regulating RhoA through atypical protein kinase C/NF-B signaling.Cancer Med. 2019 May;8(5):2288-2302. doi: 10.1002/cam4.2063. Epub 2019 Mar 7.
• [12] Quantitative Measurement of PARD3 Copy Number Variations in Human Neural Tube Defects.Cell Mol Neurobiol. 2018 Apr;38(3):605-614. doi: 10.1007/s10571-017-0506-0. Epub 2017 Jun 16.
• [13] Expression of Par3 polarity protein correlates with poor prognosis in ovarian cancer.BMC Cancer. 2016 Nov 17;16(1):897. doi: 10.1186/s12885-016-2929-2.
• [14] Protease-activated receptors (PAR)-1 and -3 drive epithelial-mesenchymal transition of alveolar epithelial cells - potential role in lung fibrosis.Thromb Haemost. 2013 Aug;110(2):295-307. doi: 10.1160/TH12-11-0854. Epub 2013 Jun 6.
• [15] PAR-1- and PAR-3-type thrombin receptor expression in primary cultures of human renal cell carcinoma cells.Int J Oncol. 2002 Jan;20(1):177-80.
• [16] Thrombin regulates the metastatic potential of human rhabdomyosarcoma cells: distinct role of PAR1 and PAR3 signaling.Mol Cancer Res. 2010 May;8(5):677-90. doi: 10.1158/1541-7786.MCR-10-0019. Epub 2010 May 4.
• [17] PARD3 Inactivation in Lung Squamous Cell Carcinomas Impairs STAT3 and Promotes Malignant Invasion.Cancer Res. 2015 Apr 1;75(7):1287-97. doi: 10.1158/0008-5472.CAN-14-2444.
• [18] MicroRNA 483-3p targets Pard3 to potentiate TGF-1-induced cell migration, invasion, and epithelial-mesenchymal transition in anaplastic thyroid cancer cells.Oncogene. 2019 Jan;38(5):699-715. doi: 10.1038/s41388-018-0447-1. Epub 2018 Aug 31.
• [19] Loss of TDP43 inhibits progression of triple-negative breast cancer in coordination with SRSF3.Proc Natl Acad Sci U S A. 2018 Apr 10;115(15):E3426-E3435. doi: 10.1073/pnas.1714573115. Epub 2018 Mar 26.
• [20] Associations with tight junction genes PARD3 and MAGI2 in Dutch patients point to a common barrier defect for coeliac disease and ulcerative colitis.Gut. 2008 Apr;57(4):463-7. doi: 10.1136/gut.2007.133132. Epub 2007 Nov 7.
• [21] Diversified Application of Barcoded PLATO (PLATO-BC) Platform for Identification of Protein Interactions.Genomics Proteomics Bioinformatics. 2019 Jun;17(3):319-331. doi: 10.1016/j.gpb.2018.12.010. Epub 2019 Sep 5.
• [22] ASPP2 suppresses invasion and TGF-1-induced epithelial-mesenchymal transition by inhibiting Smad7 degradation mediated by E3 ubiquitin ligase ITCH in gastric cancer.Cancer Lett. 2017 Jul 10;398:52-61. doi: 10.1016/j.canlet.2017.04.002. Epub 2017 Apr 9.
• [23] Activated protein C protects from GvHD via PAR2/PAR3 signalling in regulatory T-cells.Nat Commun. 2017 Aug 21;8(1):311. doi: 10.1038/s41467-017-00169-4.
• [24] Elevated expression of Par3 promotes prostate cancer metastasis by forming a Par3/aPKC/KIBRA complex and inactivating the hippo pathway.J Exp Clin Cancer Res. 2017 Oct 10;36(1):139. doi: 10.1186/s13046-017-0609-y.
• [25] Elevated leptin disrupts epithelial polarity and promotes premalignant alterations in the mammary gland.Oncogene. 2019 May;38(20):3855-3870. doi: 10.1038/s41388-019-0687-8. Epub 2019 Jan 22.
• [26] Thrombin promotes matrix metalloproteinase-13 expression through the PKC c-Src/EGFR/PI3K/Akt/AP-1 signaling pathway in human chondrocytes.Mediators Inflamm. 2013;2013:326041. doi: 10.1155/2013/326041. Epub 2013 Dec 9.
• [27] Mutational landscape of aggressive cutaneous squamous cell carcinoma.Clin Cancer Res. 2014 Dec 15;20(24):6582-92. doi: 10.1158/1078-0432.CCR-14-1768. Epub 2014 Oct 10.
• [28] The epidermal polarity protein Par3 is a non-cell autonomous suppressor of malignant melanoma.J Exp Med. 2017 Feb;214(2):339-358. doi: 10.1084/jem.20160596. Epub 2017 Jan 17.
• [29] Rare Deleterious PARD3 Variants in the aPKC-Binding Region are Implicated in the Pathogenesis of Human Cranial Neural Tube Defects Via Disrupting Apical Tight Junction Formation.Hum Mutat. 2017 Apr;38(4):378-389. doi: 10.1002/humu.23153. Epub 2017 Feb 15.
• [30] A single-copy Sleeping Beauty transposon mutagenesis screen identifies new PTEN-cooperating tumor suppressor genes.Nat Genet. 2017 May;49(5):730-741. doi: 10.1038/ng.3817. Epub 2017 Mar 20.
• [31] Loss of Par3 promotes prostatic tumorigenesis by enhancing cell growth and changing cell division modes.Oncogene. 2019 Mar;38(12):2192-2205. doi: 10.1038/s41388-018-0580-x. Epub 2018 Nov 22.
• [32] Integrative omics data analyses of repeated dose toxicity of valproic acid in vitro reveal new mechanisms of steatosis induction. Toxicology. 2018 Jan 15;393:160-170.
• [33] Integrating multiple omics to unravel mechanisms of Cyclosporin A induced hepatotoxicity in vitro. Toxicol In Vitro. 2015 Apr;29(3):489-501.
• [34] Transcriptional and Metabolic Dissection of ATRA-Induced Granulocytic Differentiation in NB4 Acute Promyelocytic Leukemia Cells. Cells. 2020 Nov 5;9(11):2423. doi: 10.3390/cells9112423.
• [35] Bringing in vitro analysis closer to in vivo: studying doxorubicin toxicity and associated mechanisms in 3D human microtissues with PBPK-based dose modelling. Toxicol Lett. 2018 Sep 15;294:184-192.
• [36] Physiological and toxicological transcriptome changes in HepG2 cells exposed to copper. Physiol Genomics. 2009 Aug 7;38(3):386-401.
• [37] Effect of nephrotoxicants and hepatotoxicants on gene expression profile in human peripheral blood mononuclear cells. Biochem Biophys Res Commun. 2010 Oct 15;401(2):245-50. doi: 10.1016/j.bbrc.2010.09.039. Epub 2010 Sep 16.
• [38] Quantitative proteomics reveals a broad-spectrum antiviral property of ivermectin, benefiting for COVID-19 treatment. J Cell Physiol. 2021 Apr;236(4):2959-2975. doi: 10.1002/jcp.30055. Epub 2020 Sep 22.
• [39] Prenatal arsenic exposure and the epigenome: identifying sites of 5-methylcytosine alterations that predict functional changes in gene expression in newborn cord blood and subsequent birth outcomes. Toxicol Sci. 2015 Jan;143(1):97-106. doi: 10.1093/toxsci/kfu210. Epub 2014 Oct 10.
• [40] Quantitative phosphoproteomics reveal cellular responses from caffeine, coumarin and quercetin in treated HepG2 cells. Toxicol Appl Pharmacol. 2022 Aug 15;449:116110. doi: 10.1016/j.taap.2022.116110. Epub 2022 Jun 7.
• [41] Transcriptional profiling of testosterone-regulated genes in the skeletal muscle of human immunodeficiency virus-infected men experiencing weight loss. J Clin Endocrinol Metab. 2007 Jul;92(7):2793-802. doi: 10.1210/jc.2006-2722. Epub 2007 Apr 17.
• [42] Exposure to Insecticides Modifies Gene Expression and DNA Methylation in Hematopoietic Tissues In Vitro. Int J Mol Sci. 2023 Mar 26;24(7):6259. doi: 10.3390/ijms24076259.
• [43] Selenium and vitamin E: cell type- and intervention-specific tissue effects in prostate cancer. J Natl Cancer Inst. 2009 Mar 4;101(5):306-20.
• [44] Comparison of HepG2 and HepaRG by whole-genome gene expression analysis for the purpose of chemical hazard identification. Toxicol Sci. 2010 May;115(1):66-79.
• [45] Endoplasmic reticulum stress and MAPK signaling pathway activation underlie leflunomide-induced toxicity in HepG2 Cells. Toxicology. 2017 Dec 1;392:11-21.
• [46] Cell-based two-dimensional morphological assessment system to predict cancer drug-induced cardiotoxicity using human induced pluripotent stem cell-derived cardiomyocytes. Toxicol Appl Pharmacol. 2019 Nov 15;383:114761. doi: 10.1016/j.taap.2019.114761. Epub 2019 Sep 15.
• [47] Expression and DNA methylation changes in human breast epithelial cells after bisphenol A exposure. Int J Oncol. 2012 Jul;41(1):369-77.
• [48] Pleiotropic combinatorial transcriptomes of human breast cancer cells exposed to mixtures of dietary phytoestrogens. Food Chem Toxicol. 2009 Apr;47(4):787-95.
• [49] Gene expression profiling of 30 cancer cell lines predicts resistance towards 11 anticancer drugs at clinically achieved concentrations. Int J Cancer. 2006 Apr 1;118(7):1699-712. doi: 10.1002/ijc.21570.