HSH2D

Summary

HSH2D (hematopoietic SH2 domain containing, HGNC:24920) is a protein-coding gene on chromosome 19p13.11, encoding Hematopoietic SH2 domain-containing protein (Q96JZ2). May be a modulator of the apoptotic response through its ability to affect mitochondrial stability.

T-cell activation requires 2 signals: recognition of antigen by the T-cell receptor (see TCR; MIM 186880) and a costimulatory signal provided primarily by CD28 (MIM 186760) in naive T cells. HSH2 is a target of both of these signaling pathways (Greene et al., 2003 [PubMed 12960172]).

Source: NCBI Gene 84941 — RefSeq curated summary.

At a glance

• GWAS associations: 3
• Clinical variants (ClinVar): 40 total
• MANE Select transcript: NM_001382417

Identifiers

Gene identifiers

Field	Value
HGNC ID	HGNC:24920
Approved symbol	HSH2D
Name	hematopoietic SH2 domain containing
Location	19p13.11
Locus type	gene with protein product
Status	Approved
Aliases	ALX, HSH2, FLJ14886
Ensembl gene	ENSG00000196684
Ensembl biotype	protein_coding
OMIM	608349
Entrez	84941

Gene structure

Transcript identifiers

Ensembl transcripts: 10 — 5 protein_coding, 4 nonsense_mediated_decay, 1 retained_intron

ENST00000535834, ENST00000586872, ENST00000591154, ENST00000593031, ENST00000613195, ENST00000613986, ENST00000616070, ENST00000616645, ENST00000874628, ENST00000912672

RefSeq mRNA: 6 — MANE Select: NM_001382417 NM_001352265, NM_001352266, NM_001369808, NM_001369809, NM_001382417, NM_032855

CCDS: CCDS74304

Canonical transcript exons

ENST00000613986 — 6 exons

Exon	Start	End
ENSE00002892503	16143685	16143774
ENSE00003473849	16153043	16153208
ENSE00003475979	16154399	16154491
ENSE00003476265	16148724	16148875
ENSE00003575154	16152552	16152641
ENSE00003711661	16157210	16158575

Expression profiles

Bgee: expression breadth ubiquitous, 164 present calls, max score 97.87.

FANTOM5 (CAGE): breadth broad, TPM avg 10.8697 / max 705.8619, expressed in 564 samples.

FANTOM5 promoters (1 alternative TSS)

Promoter ID	TPM avg	Samples expressed
174388	10.8697	564

Top tissues by expression

240 total, by Bgee expression score (0-100, higher = more expressed):

Tissue	Anatomy ID	Expression score	Quality
granulocyte	CL:0000094	97.87	gold quality
bone marrow cell	CL:0002092	94.89	gold quality
leukocyte	CL:0000738	93.42	gold quality
monocyte	CL:0000576	93.36	gold quality
vermiform appendix	UBERON:0001154	92.82	gold quality
spleen	UBERON:0002106	92.74	gold quality
blood	UBERON:0000178	92.14	gold quality
lymph node	UBERON:0000029	89.62	gold quality
mucosa of transverse colon	UBERON:0004991	86.69	gold quality
rectum	UBERON:0001052	86.19	gold quality
small intestine Peyer’s patch	UBERON:0003454	86.13	gold quality
caecum	UBERON:0001153	84.23	gold quality
bone marrow	UBERON:0002371	83.43	gold quality
small intestine	UBERON:0002108	83.05	gold quality
gall bladder	UBERON:0002110	80.75	gold quality
transverse colon	UBERON:0001157	79.83	gold quality
minor salivary gland	UBERON:0001830	78.14	gold quality
upper lobe of left lung	UBERON:0008952	77.81	gold quality
right uterine tube	UBERON:0001302	77.59	gold quality
tonsil	UBERON:0002372	77.48	gold quality
body of stomach	UBERON:0001161	77.09	gold quality
olfactory segment of nasal mucosa	UBERON:0005386	76.84	gold quality
male germ line stem cell (sensu Vertebrata) in testis	CL:0000089 ∩ UBERON:0000473	76.51	gold quality
upper lobe of lung	UBERON:0008948	75.94	gold quality
body of pancreas	UBERON:0001150	75.59	gold quality
mouth mucosa	UBERON:0003729	74.75	gold quality
stomach	UBERON:0000945	74.46	gold quality
right lung	UBERON:0002167	74.01	gold quality
colonic epithelium	UBERON:0000397	73.93	gold quality
saliva-secreting gland	UBERON:0001044	73.69	gold quality

Single-cell (SCXA)

Detected in 7 experiment(s), a significant marker in 7.

Experiment	Marker?	Max mean expression
E-MTAB-9067	yes	326.74
E-HCAD-6	yes	140.53
E-MTAB-8410	yes	25.51
E-ANND-3	yes	24.41
E-HCAD-11	yes	18.34
E-CURD-112	yes	11.80
E-MTAB-10553	yes	8.22

Regulation

Is transcription factor: no

miRNA regulators (miRDB)

16 targeting HSH2D, top 30 by miRDB confidence (max_score; target_count = how many genes the miRNA targets in total — lower means more specific):

miRNA	Max score	Avg score	miRNA target_count
HSA-MIR-185-3P	99.95	67.01	1743
HSA-MIR-3910	99.95	71.13	2227
HSA-MIR-4778-3P	99.93	70.40	1818
HSA-MIR-153-5P	99.89	73.86	6317
HSA-MIR-3151-5P	99.86	63.83	1069
HSA-MIR-4502	99.65	66.99	1021
HSA-MIR-5683	99.36	68.59	2083
HSA-MIR-7151-3P	99.04	69.72	2370
HSA-MIR-1227-5P	98.65	65.32	1549
HSA-MIR-676-5P	98.49	68.87	1492
HSA-MIR-6784-3P	98.39	64.88	662
HSA-MIR-5691	98.23	67.02	1335
HSA-MIR-6805-3P	98.23	67.02	1334
HSA-MIR-6862-3P	97.92	64.86	531
HSA-MIR-1288-3P	96.86	66.95	536
HSA-MIR-10392-3P	88.79	61.83	122

Literature-anchored findings (GeneRIF, showing 10)

• involved in tyrosine kinase signaling in hematopoietic cells (PMID:11700021)
• ALX is an adaptor involved in T cell activation and interleukin-2 (IL-2) promoter activation (PMID:12960172)
• the ALX SH2 domain plays a critical role in ALX function downstream of CD28 (PMID:15284240)
• ALX exerts its effect on IL-2 up-regulation in the cytoplasm and responds to TCR and CD28 signaling (PMID:16169852)
• results indicate that RvD1 specifically interacts with both ALX and GPR32 on phagocytes. (PMID:20080636)
• although ALX is inducibly phosphorylated upon TCR/CD28 stimulation, this phosphorylation is not required for ALX to inhibit T cell activation. (PMID:20471366)
• LXA decreased IgM and IgG production on activated human B cells through ALX/FPR2-dependent signaling. (PMID:24166736)
• pro-inflammatory stimuli lead to FPR2/ALX expression while LXA4 induces an anti-inflammatory response (PMID:26248046)
• Both ALX and ChemR23 were present in human synovium and medial tibial plateau bone obtained following total knee replacement surgery for osteoarthritis. (PMID:27860453)
• HSH2D contributes to methotrexate resistance in human Tcell acute lymphoblastic leukaemia. (PMID:33000278)

Cross-species orthologs

5 orthologs

Organism	Symbol	Gene ID
danio_rerio	zgc:92242	ENSDARG00000029443
danio_rerio	hsh2d	ENSDARG00000042908
mus_musculus	Hsh2d	ENSMUSG00000062007
rattus_norvegicus	Hsh2d	ENSRNOG00000023614
caenorhabditis_elegans		WBGENE00008733

Paralogs (4): SH2D2A (ENSG00000027869), SH2D4A (ENSG00000104611), SH2D4B (ENSG00000178217), SH2D7 (ENSG00000183476)

Protein

Protein identifiers

Hematopoietic SH2 domain-containing protein — Q96JZ2 (reviewed: Q96JZ2)

Alternative names: Adaptor in lymphocytes of unknown function X

All UniProt accessions (6): A0A087WT88, A0A087X0V9, Q96JZ2, K7EP18, K7ER77, K7ES08

UniProt curated annotations — full annotation on UniProt →

Function. May be a modulator of the apoptotic response through its ability to affect mitochondrial stability. Adapter protein involved in tyrosine kinase and CD28 signaling. Seems to affect CD28-mediated activation of the RE/AP element of the interleukin-2 promoter.

Subunit / interactions. Interacts with FES and TNK2.

Subcellular location. Cytoplasm. Nucleus.

Tissue specificity. Predominantly expressed in spleen and hematopoietic cells such as peripheral blood leukocytes and weakly expressed in prostate, thymus, heart, small intestine and placenta.

Post-translational modifications. May be phosphorylated by FES and ACK1.

Isoforms (2)

UniProt ID	Names	Canonical?
Q96JZ2-1	1	yes
Q96JZ2-2	2

RefSeq proteins (6): NP_001339194, NP_001339195, NP_001356737, NP_001356738, NP_001369346, NP_116244 (=MANE)

Domains & families (InterPro)

ID	Name	Type
IPR000980	SH2	Domain
IPR035047	HSH2D_SH2	Domain
IPR036860	SH2_dom_sf	Homologous_superfamily

Pfam: PF00017

UniProt features (25 total): mutagenesis site 7, strand 7, compositionally biased region 3, helix 2, region of interest 2, chain 1, domain 1, sequence conflict 1, splice variant 1

Structure

Experimental structures (PDB)

1 structures.

PDB	Method	Resolution (Å)
2CS0	SOLUTION NMR

Predicted structure (AlphaFold)

Model	pLDDT	Fraction very-high
AF-Q96JZ2-F1	62.94	0.19

Functional residue map

Curated UniProt residues grouped by drug-discovery relevance — catalytic, ligand-binding, modification, and mutation-validated positions. Source: UniProtKB sequence features.

Mutagenesis-validated functional residues (7):

Position	Phenotype
116	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
135	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
192–195	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
341	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
346–349	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
10–13	no change in the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.
59	loss of the ability to inhibit re/ap up-regulation in response to tcr/cd28 stimulation.

Function

Pathways and Gene Ontology

Reactome pathways

0 pathways

MSigDB gene sets: 148 (showing top): GOBP_MEMBRANE_DEPOLARIZATION, GOBP_REGULATION_OF_LEUKOCYTE_APOPTOTIC_PROCESS, GOZGIT_ESR1_TARGETS_DN, GOBP_REGULATION_OF_LYMPHOCYTE_APOPTOTIC_PROCESS, GOBP_REGULATION_OF_MEMBRANE_DEPOLARIZATION, CAGCTG_AP4_Q5, GOBP_NEGATIVE_REGULATION_OF_LEUKOCYTE_APOPTOTIC_PROCESS, GOBP_REGULATION_OF_B_CELL_APOPTOTIC_PROCESS, GOBP_NEGATIVE_REGULATION_OF_LYMPHOCYTE_APOPTOTIC_PROCESS, GOBP_LEUKOCYTE_APOPTOTIC_PROCESS, BERTUCCI_MEDULLARY_VS_DUCTAL_BREAST_CANCER_UP, GOBP_REGULATION_OF_MITOCHONDRIAL_MEMBRANE_POTENTIAL, JAATINEN_HEMATOPOIETIC_STEM_CELL_UP, GOBP_LYMPHOCYTE_APOPTOTIC_PROCESS, YNGTTNNNATT_UNKNOWN

GO Biological Process (4): negative regulation of B cell apoptotic process (GO:0002903), signal transduction (GO:0007165), T cell activation (GO:0042110), negative regulation of mitochondrial depolarization (GO:0051902)

GO Molecular Function (2): protein-macromolecule adaptor activity (GO:0030674), protein binding (GO:0005515)

GO Cellular Component (4): nucleus (GO:0005634), cytoplasm (GO:0005737), mitochondrion (GO:0005739), cytosol (GO:0005829)

GO top-level categories

Rollup of top GO terms by namespace:

Category	Terms
intracellular membrane-bounded organelle	2
cellular anatomical structure	2
cytoplasm	2
B cell apoptotic process	1
regulation of B cell apoptotic process	1
negative regulation of lymphocyte apoptotic process	1
cell communication	1
cellular process	1
signaling	1
regulation of cellular process	1
cellular response to stimulus	1
lymphocyte activation	1
mitochondrial depolarization	1
regulation of mitochondrial depolarization	1
negative regulation of membrane depolarization	1
protein binding	1
molecular adaptor activity	1
binding	1
intracellular anatomical structure	1

Protein interactions and networks

STRING

888 interactions, top by confidence (×1000):

Protein A	Protein B	Partner UniProt	Score
HSH2D	FPR2	P25090	845
HSH2D	ANXA1	P04083	837
HSH2D	GPR32	O75388	822
HSH2D	TNK2	Q07912	768
HSH2D	CMKLR1	Q99788	745
HSH2D	LTB4R	Q15722	726
HSH2D	ALB	P02768	571
HSH2D	CD28	P10747	536
HSH2D	GPR18	Q14330	528
HSH2D	A0A087WTN9	A0A087WTN9	501
HSH2D	CDC42	P21181	473
HSH2D	VWF	P04275	471
HSH2D	IL6R	P08887	471
HSH2D	CD4	P01730	467
HSH2D	KANSL3	Q9P2N6	464

IntAct

34 interactions, top by confidence:

A	B	Type	Score
HSH2D	TSG101	psi-mi:“MI:0915”(physical association)	0.560
HSH2D	LDHAL6B	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	PINK1	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	RBP7	psi-mi:“MI:0915”(physical association)	0.490
CRK	HSH2D	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	TSC1	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	PIK3R3	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	CNMD	psi-mi:“MI:0915”(physical association)	0.490
HSH2D		psi-mi:“MI:0915”(physical association)	0.490
RBP7	HSH2D	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	CRK	psi-mi:“MI:0915”(physical association)	0.490
TSC1	HSH2D	psi-mi:“MI:0915”(physical association)	0.490
PIK3R3	HSH2D	psi-mi:“MI:0915”(physical association)	0.490
	HSH2D	psi-mi:“MI:0915”(physical association)	0.490
HSH2D	AR	psi-mi:“MI:0407”(direct interaction)	0.440
HSH2D	GAB1	psi-mi:“MI:0407”(direct interaction)	0.440
HSH2D	KIT	psi-mi:“MI:0407”(direct interaction)	0.440
HSH2D	MET	psi-mi:“MI:0407”(direct interaction)	0.440
HSH2D	NSA2	psi-mi:“MI:0915”(physical association)	0.400
OLIG1	HSH2D	psi-mi:“MI:0915”(physical association)	0.370
TNK2	HSH2D	psi-mi:“MI:0915”(physical association)	0.370
HSH2D	MPP3	psi-mi:“MI:0915”(physical association)	0.370
HSH2D	ALK	psi-mi:“MI:0915”(physical association)	0.370

BioGRID (30): HSH2D (Two-hybrid), LDHAL6B (Two-hybrid), PPAPDC2 (Two-hybrid), RBP7 (Two-hybrid), LECT1 (Two-hybrid), PIK3R3 (Two-hybrid), PINK1 (Two-hybrid), TSC1 (Two-hybrid), OLIG1 (Two-hybrid), TNK2 (Two-hybrid), FES (Two-hybrid), NSA2 (Affinity Capture-MS), HSH2D (Affinity Capture-MS), HSH2D (Two-hybrid), HSH2D (Reconstituted Complex)

ESM2 similar proteins: A1L3T7, A2CI98, A2CJ06, A2VCK2, D3ZQL6, E1BBG2, M3WHG5, O15037, O43918, O70303, O75333, P48778, P59729, Q32PJ7, Q3KP66, Q3T191, Q5JYT7, Q5T124, Q5U4F0, Q5XIS1, Q6AXX1, Q6VYH9, Q7TSI1, Q80U38, Q8BGT6, Q8BHW9, Q8BWG4, Q8BWQ5, Q8BZ33, Q8BZW2, Q8C0J6, Q8C2K5, Q8CFK6, Q8IV53, Q8IY33, Q8IZW8, Q8K1S6, Q8K330, Q8N3F8, Q8TB24

Diamond homologs: A6NKC9, A6X942, G5ECJ6, O08908, O88834, P03949, P46109, P47941, Q00655, Q08012, Q08CX2, Q56A36, Q5SQS7, Q5U2U2, Q6AYC8, Q6VYH9, Q6YKA8, Q8BI17, Q8UUU2, Q96JZ2, Q9D7V1, Q9H788, Q9NP31, Q9QXK9, P00519, P00520, P00521, P10447, P20936, P29353, P32577, P41239, P41240, P41241, P42684, P50904, P98083, Q03160, Q0IIE2, Q0VBZ0

SIGNOR signaling

0 interactions.

Enriched among interaction partners

Reactome pathways and GO biological processes over-represented among this gene’s 19 IntAct physical interaction partners (hypergeometric vs the genome-wide background, BH-FDR, gene-set size 15–500, ranked by fold). A functional readout of the neighbourhood — distinct from this gene’s own memberships above, and biased toward well-studied / hub proteins, so read it as themes rather than proof.

Reactome pathways:

Pathway	Partners	Fold	FDR
Constitutive Signaling by Aberrant PI3K in Cancer	5	48.8×	7e-06
PI5P, PP2A and IER3 Regulate PI3K/AKT Signaling	5	37.2×	9e-06
PIP3 activates AKT signaling	5	25.7×	3e-05
Signaling by Receptor Tyrosine Kinases	5	19.9×	1e-04

Disease & clinical

Clinical variants and AI predictions

ClinVar

40 variants total. Per-class counts are floors (≥ shown; pagination cap):

Classification	Count (floor)
Pathogenic	0
Likely pathogenic	0
Uncertain significance	34
Likely benign	0
Benign	2

Top pathogenic / likely-pathogenic (0)

SpliceAI

1463 predictions. Top by Δscore:

Variant	Effect	Δscore
19:16143626:CCCCA:C	acceptor_loss	1.0000
19:16143627:CCCAG:C	acceptor_loss	1.0000
19:16143630:AG:A	acceptor_gain	1.0000
19:16143631:G:T	acceptor_loss	1.0000
19:16143631:GG:G	acceptor_gain	1.0000
19:16148872:GAGA:G	donor_gain	1.0000
19:16148874:GA:G	donor_gain	1.0000
19:16148876:G:GG	donor_gain	1.0000
19:16152550:AGG:A	acceptor_gain	1.0000
19:16152551:GGG:G	acceptor_gain	1.0000
19:16152642:G:GG	donor_gain	1.0000
19:16153181:G:GT	donor_gain	1.0000
19:16154397:A:AG	acceptor_gain	1.0000
19:16154397:AGAAG:A	acceptor_gain	1.0000
19:16154398:G:GG	acceptor_gain	1.0000
19:16154398:GAAGG:G	acceptor_gain	1.0000
19:16154488:GGAG:G	donor_gain	1.0000
19:16154489:G:GT	donor_gain	1.0000
19:16154489:GAG:G	donor_gain	1.0000
19:16154489:GAGGT:G	donor_loss	1.0000
19:16154492:G:GC	donor_loss	1.0000
19:16154492:G:GG	donor_gain	1.0000
19:16154493:T:A	donor_loss	1.0000
19:16143630:A:AG	acceptor_gain	0.9900
19:16143630:AGG:A	acceptor_gain	0.9900
19:16143631:G:GA	acceptor_gain	0.9900
19:16143631:GGG:G	acceptor_gain	0.9900
19:16143631:GGGCT:G	acceptor_gain	0.9900
19:16143775:GT:G	donor_loss	0.9900
19:16143776:T:G	donor_loss	0.9900

AlphaMissense

2288 scored. Top likely-pathogenic:

Variant	Protein change	am_pathogenicity
19:16152592:T:C	F56L	0.996
19:16152594:T:A	F56L	0.996
19:16152594:T:G	F56L	0.996
19:16148850:T:A	W34R	0.993
19:16148850:T:C	W34R	0.993
19:16152607:A:C	S61R	0.992
19:16152609:T:A	S61R	0.992
19:16152609:T:G	S61R	0.992
19:16152556:G:C	A44P	0.990
19:16157774:T:C	F347L	0.989
19:16157776:T:A	F347L	0.989
19:16157776:T:G	F347L	0.989
19:16148854:T:C	F35S	0.985
19:16152603:G:C	R59S	0.985
19:16152603:G:T	R59S	0.985
19:16148853:T:C	F35L	0.984
19:16148855:C:A	F35L	0.984
19:16148855:C:G	F35L	0.984
19:16152593:T:C	F56S	0.984
19:16152602:G:T	R59M	0.984
19:16148802:T:C	F18L	0.981
19:16148804:T:A	F18L	0.981
19:16148804:T:G	F18L	0.981
19:16153064:T:A	H79Q	0.980
19:16153064:T:G	H79Q	0.980
19:16152625:T:G	Y67D	0.979
19:16153144:T:A	V106D	0.979
19:16148852:G:C	W34C	0.978
19:16148852:G:T	W34C	0.978
19:16152602:G:C	R59T	0.978

dbSNP variants (sampled 300 via entrez): RS1000042614 (19:16136411 C>A,T), RS1000476877 (19:16135448 T>C), RS1000521792 (19:16146470 G>A), RS1000581144 (19:16142606 G>C), RS1000674344 (19:16158154 C>A), RS1000749660 (19:16156933 C>A), RS1000759008 (19:16137187 C>T), RS1001212577 (19:16136919 T>C), RS1001275524 (19:16158242 C>G,T), RS1001490429 (19:16140384 C>A), RS1001586315 (19:16146341 G>A), RS1001638712 (19:16146041 A>G), RS1001663927 (19:16134849 G>C), RS1001752474 (19:16151975 C>T), RS1001824029 (19:16140584 G>A)

Disease associations

OMIM: gene MIM:608349 | disease phenotypes:

GenCC curated gene-disease

Mondo (0):

Orphanet (0):

HPO phenotypes

0 total (0 of 0 shown, HPO-id order):

GWAS associations

3 associations (top):

Study	Trait	p-value
GCST90002390_532	Mean corpuscular hemoglobin	1.000000e-28
GCST90002392_71	Mean corpuscular volume	7.000000e-22
GCST90002397_192	Mean spheric corpuscular volume	2.000000e-09

EFO canonical traits (1, from GWAS)

EFO ID	Trait name
EFO:0004527	mean corpuscular hemoglobin

Drugs & pharmacology

Drug and pharmacology data

Is drug target: no

PharmGKB: 1 entry (VIP=true, CPIC=false)

CTD chemical–gene interactions

28 total (human), top 28 by PubMed support.

Chemical	Actions (top 5)	PubMed papers
(+)-JQ1 compound	decreases expression	2
Benzo(a)pyrene	affects methylation	2
Estradiol	affects cotreatment, decreases expression	2
Nickel	increases expression	2
Tobacco Smoke Pollution	decreases expression, affects expression	2
aristolochic acid I	increases expression	1
triphenyl phosphate	affects expression	1
ethyl-p-hydroxybenzoate	decreases expression	1
perfluorooctanoic acid	decreases expression	1
benzo(e)pyrene	decreases methylation	1
pinostrobin	increases expression	1
abrine	decreases expression	1
Acetaminophen	increases expression	1
Air Pollutants	increases abundance, increases expression	1
Cadmium	decreases expression	1
Calcitriol	increases expression, affects cotreatment	1
Drugs, Chinese Herbal	increases expression	1
Formaldehyde	decreases expression	1
Methapyrilene	decreases methylation	1
Methotrexate	increases expression	1
Methyl Methanesulfonate	decreases expression	1
Naphthoquinones	increases expression	1
Smoke	decreases expression	1
Testosterone	affects cotreatment, increases expression	1
Tretinoin	increases expression	1
Aflatoxin B1	decreases expression	1
Cadmium Chloride	decreases expression	1
Particulate Matter	increases abundance, increases expression	1

Clinical trials (associated diseases)

0 trials via MONDO — disease-level, not drug-specific.