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ID PMID Title PublicationDate abstract
23285494 Indocyanine green derivative 02-Glu-c(RGDyK)(2). 2004 Optical fluorescence imaging is increasingly used to monitor biological functions of specific targets in small animals (1-3). However, the intrinsic fluorescence of biomolecules poses a problem when fluorophores that absorb visible light (350–700 nm) are used. Near-infrared (NIR) fluorescence (650–900 nm) detection avoids the natural background fluorescence interference of biomolecules, providing a high contrast between target and background tissues. NIR fluorophores have wider dynamic range and minimal background fluorescence as a result of reduced scattering compared with visible fluorescence detection. They also have high sensitivity, resulting from low background fluorescence, and high extinction coefficients, which provide high quantum yields. The NIR region is also compatible with solid-state optical components, such as diode lasers and silicon detectors. NIR fluorescence imaging is a noninvasive complement to radionuclide imaging in small animals or with probes in close proximity to the target in humans (4). Among the various optical imaging agents, only indocyanine green (ICG), with NIR fluorescence absorption at 780 nm and emission at 820 nm, is approved by the United States Food and Drug Administration for clinical applications in angiography, blood flow evaluation, and liver function assessment (5-8). It is also under evaluation in several clinical trials for other applications, such as optical imaging and mapping of both the lymphatic vessels and lymph nodes in cancer patients for surgical dissection of tumor cells and endoscopic imaging of the pancreas and colon. Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (9). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (10-17). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. The peptide sequence Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). The α(v)β(3) antagonists are being studied as anti-tumor and anti-angiogenic agents (12, 16, 18). Various radiolabeled (such as (18)F, (64)Cu, (68)Ga, and (99m)Tc) and NIR fluorescence-labeled (such as Cy5.5, Cy7, and Cypate) RGD peptides have been introduced for imaging of tumors and tumor angiogenesis (19). ICG derivative 02 (ICG-Der-02), a hydrophilic dye, contains one carboxyl functional group for covalent conjugation to the amino group of biomolecules. Cao et al. (20) conjugated ICG-Der-02 via the α-amino group of Glu residue of Glu-c(RGDyK)(2) (dimer) peptide to form ICG-Der-02-c(RGDyK)(2.) ICG-Der-02 was also conjugated to the Ɛ-amino group of the lysine residue of RGD (linear) and c(RGDyK) (monomer) to form ICG-Der-02-RGD and ICG-Der-02-c(RGDyK), respectively. The three ICD-Der-02-labeled conjugates were evaluated for in vivo NIR optical imaging in tumor-bearing mice.
23193614 (64)Cu-1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid-Arg-rich Cys knot scaffold 2004 Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (1). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (2-7). Expression of α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. α(v)β(3) antagonists are being studied as antitumor and antiangiogenic agents, and the agonists are being studied as angiogenic agents for coronary angiogenesis (6, 8, 9). A tripeptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3) and α(v)β(6). Various radiolabeled antagonists have been introduced for imaging of tumors and tumor angiogenesis (10). Integrin α(v)β(6) plays an important role in the development of epithelial cells and is nearly undetectable on adult normal tissues. However, the levels of α(v)β(6) integrin can be upregulated during tissue remodeling and wound healing (11). On the other hand, α(v)β(6) integrin is strongly expressed on tumor cells of the oral cavity, pancreas, breast, ovary, colon, and stomach (12-14). α(v)β(6) integrin affects tumor growth, tumor invasiveness, and metastasis (13). α(v)β(6) binds to the RGD motif in fibronectin, tenascin, and the viral protein 1 (VP1) of foot-and-mouth disease virus (FMDV) (15). FMDV binds to cells through the RGD motif of the GH loop of the VP1. A consensus α(v)β(6)-binding motif DLXXL was identified by using phage display screening with minimal binding to α(v)β(3), α(IIb)β(3), and α(v)β(5) (16). Engineered cysteine knot peptides (knots) comprise a rigid molecular scaffold of ~4 kDa with three disulfide bonds and a centrally located β sheet. Kimura et al. (17) grafted a α(v)β(6)-binding peptide (RSLARTDLDHLRGR) into the loop 1 of an Arg-rich knot to produce the knot known as R(0)1, which was radiolabeled with (64)Cu via 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) conjugation to form (64)Cu-DOTA-R(0)1 as a positron emission tomography (PET) probe for in vivo imaging of α(v)β(6) integrin in tumor-bearing nude mice.
23193623 (64)Cu-1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid-Ser-rich Cys knot scaffold 2004 Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (1). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (2-7). Expression of α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. α(v)β(3) antagonists are being studied as antitumor and antiangiogenic agents, and the agonists are being studied as angiogenic agents for coronary angiogenesis (6, 8, 9). A tripeptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3) and α(v)β(6). Various radiolabeled antagonists have been introduced for imaging of tumors and tumor angiogenesis (10). Integrin α(v)β(6) plays an important role in the development of epithelial cells and is nearly undetectable on adult normal tissues. However, the levels of α(v)β(6) integrin can be upregulated during tissue remodeling and wound healing (11). On the other hand, α(v)β(6) integrin is strongly expressed on tumor cells of the oral cavity, pancreas, breast, ovary, colon, and stomach (12-14). α(v)β(6) integrin affects tumor growth, tumor invasiveness, and metastasis (13). α(v)β(6) binds to the RGD motif in fibronectin, tenascin, and the viral protein 1 (VP1) of foot-and-mouth disease virus (FMDV) (15). FMDV binds to cells through the RGD motif of the GH loop of the VP1. A consensus α(v)β(6)-binding motif DLXXL was identified by using phage display screening with minimal binding to α(v)β(3), α(IIb)β(3), and α(v)β(5) (16). Engineered cysteine knot peptides (knots) comprise a rigid molecular scaffold of ~4 kDa with three disulfide bonds and a centrally located β sheet. Kimura et al. (17) grafted a α(v)β(6)-binding peptide (RSLARTDLDHLRGR) into the loop 1 of a Ser-rich knot to produce the knot known as S(0)2, which was radiolabeled with (64)Cu via 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) conjugation to form (64)Cu-DOTA-S(0)2 as a positron emission tomography (PET) probe for in vivo imaging of α(v)β(6) integrin in tumor-bearing nude mice.
20641746 (68)Ga-1,4,7-Triazacyclononane-1,4-7-triacetic acid-Glu-c(RGDyK)-bombesin[7-14]. 2004 The amphibian bombesin (BBN or BN, a peptide of 14 amino acids) is an analog of human gastrin-releasing peptide (GRP, a peptide of 27 amino acids) that binds to GRP receptors (GRPR) with high affinity and specificity (1, 2). Both GRP and BBN share an amidated C-terminus sequence homology of seven amino acids, Trp-Ala-Val-Gly-His-Leu-Met-NH(2). BBN-Like peptides have been shown to induce various biological responses in diverse tissues, including the central nervous system (CNS) and the gastrointestinal (GI) system. They also act as potential growth factors for both normal and neoplastic tissues (3). Specific BBN receptors (BBN-R) have been identified on CNS and GI tissues and on a number of tumor cell lines (4). The BBN-R superfamily includes at least four different subtypes, namely the GRPR subtype (BB2), the neuromedin B (NMB) receptor subtype (BB1), the BB3 subtype, and the BB4 subtype. The findings of GRPR overexpression in various human tumors, such as breast, prostate, lung, colon, ovarian, and pancreatic cancers, provide opportunities for tumor imaging by designing specific molecular imaging agents to target the GRPR (5, 6). Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (7). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (8-13). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. A peptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various ligands have been introduced for imaging of tumors and tumor angiogenesis (14). Because breast and prostate cancers express GRPR and α(v)β(3), Liu et al. (15) designed an RGD-BBN heterodimer in which BBN[7-14] and c(RGDyK) are connected with a glutamate linker (BBN on the Glu side chain γ-carboxylate group and RGD on the Glu side chain α-carboxylate group). A spacer, 11-amino-3,6,9-trioxaundecanoic acid (PEG(3)), was put onto the glutamate α-amino group of Glu-RGD-BBN to increase the hydrophilicity and to relieve the steric hindrance. N-Succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB) was used to synthesize [(18)F]FB-PEG(3)-RGD-BBN for tumor targeting. Liu et al. (16) used 1,4,7-triazacyclononane-1,4-7-triacetic acid (NOTA) as a bifunctional chelator for labeling RGD-BBN to form (68)Ga-NOTA-RGD-BBN for positron emission tomography (PET) imaging of α(v)β(3) and GRPR in nude mice bearing human tumors (15, 17).
20641486 Gadolinium diethylenetriamine pentaacetic acid-Arg-Gly-Asp peptidomimetic. 2004 Magnetic resonance imaging (MRI) maps information about tissues spatially and functionally. Protons (hydrogen nuclei) are widely used in imaging because of their abundance in water molecules. Water comprises ~80% of most soft tissue. The contrast of proton MRI depends primarily on the density of the nucleus (proton spins), the relaxation times of the nuclear magnetization (T1, longitudinal, and T2, transverse), the magnetic environment of the tissues, and the blood flow to the tissues. However, insufficient contrast between normal and diseased tissues requires the development of contrast agents. Most contrast agents affect the T1 and T2 relaxation times of the surrounding nuclei, mainly the protons of water. T2* is the spin–spin relaxation time composed of variations from molecular interactions and intrinsic magnetic heterogeneities of tissues in the magnetic field (1). Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (2). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (3-8). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. The α(v)β(3) antagonists are being studied as antitumor and antiangiogenic agents, and the agonists are being studied as angiogenic agents for coronary angiogenesis (7, 9, 10). A tripeptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various radiolabeled antagonists have been introduced for imaging of tumors and tumor angiogenesis (11). Gadolinium (Gd), a lanthanide metal ion with seven unpaired electrons, has been shown to be very effective in enhancing proton relaxation because of its high magnetic moment and water coordination (12, 13). Gd-Labeled diethylenetriamine pentaacetic acid (Gd-DTPA) was the first intravenous MRI contrast agent used clinically, and a number of similar Gd chelates have been developed in an effort to further improve clinical use. However, these low molecular weight Gd chelates have short blood and tissue retention times, which limit their use as imaging agents in the vasculature and cancer. Various macromolecular Gd complexes have demonstrated superior contrast enhancement for MRI of the vasculature and carcinomas (14-16); however, these Gd complexes did not proceed into further clinical development because of high tissue accumulation and slow excretion of toxic Gd ions. Furthermore, they are largely nonspecific. A low molecular weight, non-peptide, RGD mimetic (g-mimRGD, 5-{N'-[5-(4-Methylpyridin-2-ylamino)pentanoyl]hydrazino}-3-(4-biphenyl)-5-oxopentanoic acid trifluoroacetate) was identified to have good affinity and selectivity for the α(v)β(3) integrin (50% inhibition concentration, 3 nM) (17). Burtea et al. (18) conjugated mimRGD to Gd-DTPA to form Gd-DTPA-g-minRGD for non-invasive in vivo MRI of α(v)β(3) integrin expression in transgenic apolipoprotein E–deficient (ApoE(-/-)) mice.
20641766 (64)Cu-Anti-human integrin α(v)β(3) monoclonal antibody. 2004 (64)Cu-Anti-human integrin α(v)β(3) monoclonal antibody ((64)Cu-DOTA-hLM609-II) is a radiolabeled antibody developed for positron emission tomography (PET) imaging of integrin α(v)β(3)−positive tumors (1). (64)Cu-DOTA-hLM609-II is labeled with (64)Cu which is a positron emitter with a half-life (t(½)) of 12.7 h. Cellular survival, invasion, and migration control embryonic development, angiogenesis, tumor metastasis, and other physiological processes. Among the molecules that regulate angiogenesis are integrins, a superfamily of cell adhesion proteins that form heterodimeric receptors for extracellular matrix (ECM) molecules (2, 3).These transmembrane glycoproteins consist of two noncovalently associated subunits, α and β (18 α- and 8 β-subunits in mammals), which are assembled into at least 24 α/β pairs. Several integrins, such as integrin α(v)β(3), have affinity for the arginine-glycine-aspartic acid (RGD) tripeptide motif, which is found in many ECM proteins. The integrin α(v)β(3) receptor is generally not found in normal tissue but is strongly expressed in vessels with increased angiogenesis, such as tumor vasculature. It is significantly upregulated in certain types of tumor cells and in almost all tumor vasculature. Increased levels of integrin α(v)β(3) expression are closely associated with increased cell invasion and metastasis. Molecular imaging of a probe that binds to the integrin α(v)β(3) can be used to image tumor vasculature and evaluate angiogenic response to tumor therapy (4, 5). Radiolabeled monoclonal antibodies (MAbs) have been developed for both the diagnosis and treatment of tumors (6-8). Radiometals with appropriate photon or positron emissions can be used to radiolabel antibodies and in imaging studies (9, 10). Cheresh and Harper (11, 12) first reported the synthesis of LM609 MAb, a murine IgG1 antibody (mLM609), which appeared to react with and recognize α(v)β(3) as one entity. The mLM609 was humanized by grafting the murine complementary-determining regions (CDRs) onto a human framework (13, 14). The humanized LM609 (hLM609) was shown to retain the affinity properties of the murine antibody. This first version of hLM609 (hLM609-I) was used in clinical studies for cancer therapy (15). These studies reported no significant toxicity or immune response from hLM609-I. However, (99m)Tc labeling of hLM609-I was unsuccessful due to the in vivo instability of the (99m)Tc label (16). Another clone (hLM609-II) with >50-fold enhanced affinity was also produced using a phage-expressed libraries and focused mutagenesis strategy in a stepwise fashion (13, 15). hLM609-II is currently in clinical trials for treatment of prostate cancer, psoriasis, melanoma, and rheumatoid arthritis. Cai et al. (1) successfully radiolabeled hLM609-II with (64)Cu using 1,4,7,10-tetra-azacyclododecane N,N',N'',N'''-tetraacetic acid (DOTA) as the bifunctional chelator.
20945564 4-[(18)F]Fluorobenzoyl-knottin 2.5D. 2004 Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (1). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (2-7). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. The α(v)β(3) antagonists are being studied as antitumor and antiangiogenic agents, and the agonists are being studied as angiogenic agents for coronary angiogenesis (6, 8, 9). The peptide sequence Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various radiolabeled antagonists have been introduced for imaging of tumors and tumor angiogenesis (10). Most cyclic RGD peptides are composed of five amino acids. Haubner et al. (11) reported that various cyclic RGD peptides exhibit selective inhibition of binding (expressed as 50% inhibitory concentration (IC(50))) to α(v)β(3) (IC(50), 7–40 nM) but not to α(v)β(5) (IC(50), 600–4,000 nM) or α(IIb)β(3) (IC(50), 700–5,000 nM) integrins. Various radiolabeled cyclic RGD peptides have been found to have high accumulation in tumors in nude mice (12). Only one cyclic RGD peptide used for imaging, [(18)F]fluoropropionyl-galacto-c(Arg-Gly-Asp-d-Phe-Lys) ([(18)F]-galacto-RGD), has been investigated for measuring expression of α(v)β(3) integrin in cancer patients with tumors. However, [(18)F]-galacto-RGD has been shown to have low tumor accumulation and low signal/background ratios. Cystine knot peptides (knottins) share a common disulfide-bonded framework and a triple-stranded β-sheet fold (13). The integrin-binding RGD motif was grafted into a knottin from trypsin inhibitor II of the squash plant (Ecballium elaterium). Knottin 2.5D (with three disulfide bonds; (GCPQGRGDWAPTSCSQDSDCLAGCVCGPNGFCG-NH(2)) was identified from a series of genetically engineered knottin peptides to have nanomolar binding to the α(v)β(3), α(v)β(5), and α(5)β(1) integrin receptors (14, 15). Knottin 2.5D was labeled with N-succinimidyl-4-[(18)F]fluorobenzoate ([(18)F]SFB) to form 4-[(18)F]fluorobenzoyl-knottin 2.5D ([(18)F]FB-2.5D) and evaluated for positron emission tomography (PET) imaging of tumor xenografts in mice (16).
20641655 (111)In-Diethylenetriaminepentaacetic acid-benzyl-succinamido-Lys-IRDye800-c(Arg-Gly-Asp-D 2004 Optical fluorescence imaging is increasingly being used to monitor biological functions of specific targets in small animals (1-3). However, the intrinsic fluorescence of biomolecules poses a problem when fluorophores that absorb visible light (350–700 nm) are used. Near-infrared (NIR) fluorescence (700–1,000 nm) detection avoids the natural background fluorescence interference of biomolecules, providing a high contrast between target and background tissues in small animals. NIR fluorophores have a wider dynamic range and minimal background fluorescence as a result of reduced scattering compared with visible fluorescence detection. NIR fluorophores also have high sensitivity, attributable to low background fluorescence, and high extinction coefficients, which provide high quantum yields. The NIR region is also compatible with solid-state optical components, such as diode lasers and silicon detectors. NIR fluorescence imaging is a non-invasive alternative to radionuclide imaging in small animals (4, 5). Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (6). Integrins consist of an α and a β subunit, and they are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (7-12). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. Antagonists of α(v)β(3) are being studied as antitumor and antiangiogenic agents and its agonists are being studied as angiogenic agents for coronary angiogenesis (11, 13, 14). A peptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various radiolabeled antagonists have been introduced for tumor imaging and tumor angiogenesis (15). Most of the cyclic RGD peptides are composed of five amino acids. Haubner et al. (16) reported that various cyclic RGD peptides exhibit selective inhibition of binding to α(v)β(3) (50% inhibition concentration (IC(50)), 7–40 nM) but not to α(v)β(5) (IC(50), 600–4,000 nM) or α(IIb)β(3) (IC(50), 700–5,000 nM) integrins. Various radiolabeled cyclic RGD peptides have been found to have high accumulation in tumors in nude mice (17). Li et al. (18) reported the development of (111)In-DTPA-Bz-SA-Lys-IRDye800-c(RGDfK), a multimodality imaging agent for imaging α(v)β(3) expression on breast tumors; single-photon emission computed tomography (SPECT) imaging used (111)In-DTPA-Bz-SA and NIR imaging used IRDye800.
20641690 (64)Cu-1,4,7,10-Tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid-polyethylene-glycol-s 2004 Optical fluorescence imaging is increasingly used to visualize biological functions of specific targets (1, 2). However, the intrinsic fluorescence of biomolecules poses a problem when fluorophores that absorb visible light (350–700 nm) are used. Near-infrared (NIR) fluorescence (700–1,000 nm) detection avoids the background fluorescence interference of natural biomolecules, providing a high contrast between target and background tissues. NIR fluorophores have wider dynamic range and minimal background as a result of reduced scattering compared with visible fluorescence detection. They also have high sensitivity, resulting from low infrared background, and high extinction coefficients, which provide high quantum yields. The NIR region is also compatible with solid-state optical components, such as diode lasers and silicon detectors. NIR fluorescence imaging is becoming a non-invasive alternative to radionuclide imaging in small animals. Carbon nanotubes are made of carbon fullerene carbon units, which respond to local dielectric changes without photobleaching (3, 4). They can be tuned in a range of wavelengths for NIR absorption, thus providing broad excitation profiles and high absorption coefficients. They can be coated and capped with hydrophilic materials for additional conjugation with biomolecules, such as peptides, antibodies, nucleic acids, and small organic compounds for in vitro and in vivo studies (5). Single-walled carbon nanotubes (SWNTs) have a diameter of 1–5 nm and a length of 200–1,000 nm, and they have been shown to be non-toxic to cells in vitro (6). However, there have been limited in vivo studies of SWNT toxicological and pharmacological profiles in small animals. Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (7). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (8-13). Expression of the α(v)β(3) integrin is strong in tumor cells and activated endothelial cells, whereas expression is weak in resting endothelial cells and most normal tissues. Antagonists of α(v)β(3) are being studied as antitumor and antiangiogenic agents and the agonists of α(v)β(3) are being studied as angiogenic agents for coronary angiogenesis (12, 14, 15). A peptide sequence consisting of Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various ligands have been introduced for imaging of tumors and tumor angiogenesis (16). Liu et al. (17) conjugated (64)Cu-1,4,7,10-tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid ((64)Cu-DOTA) and c(RGDyK) to phospholipid-polyethylene-glycol (PL-PEG)–coated SWNTs ((64)Cu-DOTA-SWNT-PEG-c(RGDyK)) for imaging α(v)β(3) integrin expression in tumors and their vasculatures.
22359781 (64)Cu-Bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane-cyclo(Arg-Gly-Asp-d-Ph 2004 Integrins are a family of heterodimeric glycoproteins on cell surfaces that mediate diverse biological events involving cell–cell and cell–matrix interactions (1). Integrins consist of an α and a β subunit and are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (2-7). Expression of the α(v)β(3) integrin is strong on tumor cells and activated endothelial cells, whereas expression is weak on resting endothelial cells and most normal tissues. The α(v)β(3) antagonists are being studied as antitumor and antiangiogenic agents and the agonists are being studied as angiogenic agents for coronary angiogenesis (6, 8, 9). The peptide sequence Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various radiolabeled antagonists have been introduced for imaging of tumors and tumor angiogenesis (10). Most of the cyclic RGD peptides are composed of five amino acids. Various cyclic RGD peptides exhibit selective inhibition of binding to α(v)β(3) (50% inhibition concentration (IC(50)), 7–40 nM) but not to α(v)β(5) (IC(50), 600–4,000 nM) or α(IIb)β(3) (IC(50), 700–5,000 nM) integrins (11). Various radiolabeled cyclic RGD peptides and peptidomimetics have been found to have high accumulation in tumors in mice (12, 13). From these developments, [(18)F]galacto-c(RGDfK) has been evaluated in a number of clinical studies for imaging of α(v)β(3) in cancer patients (14-18). Knetsch et al. (19) reported the development of (68)Ga-1,4,7-triazacyclononane,1-glutaric acid-4,7-acetic acid-cyclo(Arg-Gly-Asp-d-Phe-Lys) ((68)Ga-NODAGA-c(RGDfK)) for positron emission tomography (PET) imaging of α(v)β(3) receptors in nude mice bearing melanoma tumors. 1-(1-Carboxy-3-carbo-tert-butoxypropyl)-4,7-(carbo-tert-butoxymethyl)-1,4,7-triazacyclononane (NODAGA(tBu)(3)) was used to prepare (68)Ga-NODAGA-c(RGDfK). Dumont et al. (20) chelated NODAGA-c(RGDfK) with (64)Cu to form (64)Cu-NODAGA-c(RGDfK) for PET imaging of α(v)β(3) receptors in nude mice bearing human glioblastoma tumors. For evaluation as a PET imaging agent for α(v)β(3), (64)Cu has been attached to c(RGDfK) via bis(carboxymethyl)-1,4,8,11-tetraazabicyclo[6.6.2]hexadecane (CB-TE2A) to form (64)Cu-CB-TE2A-c(RGDfK). The CB-TE2A chelating agent is thought to be a more stable chelate for Cu.
21938857 Cys-Asp-Cys-Arg-Gly-Asp-Cys-Phe-Cys/Cy5.5-Ferritin (64)Cu-loaded nanocages. 2004 Optical fluorescence imaging is increasingly used to monitor biological functions of specific targets in small animals (1-3). However, the intrinsic fluorescence of biomolecules poses a problem when fluorophores that absorb visible light (350–700 nm) are used. Near-infrared (NIR) fluorescence (700–1,000 nm) detection avoids the background fluorescence interference of natural biomolecules, providing a high contrast between target and background tissues. NIR fluorophores have a wider dynamic range and minimal background as a result of reduced scattering compared with visible fluorescence detection. They also have high sensitivity, resulting from low fluorescence background, and high extinction coefficients, which provide high quantum yields. The NIR region is also compatible with solid-state optical components, such as diode lasers and silicon detectors. NIR fluorescence (NIRF) imaging is becoming a noninvasive alternative to radionuclide imaging in small animals (4, 5). Integrins are a family of heterodimeric, cell-surface glycoproteins that mediate diverse biological events involving cell–cell and cell–matrix interactions (6). Integrins comprise an α and a β subunit, and they are important for cell adhesion and signal transduction. The α(v)β(3) integrin is the most prominent receptor class, affecting tumor growth, tumor invasiveness, metastasis, tumor-induced angiogenesis, inflammation, osteoporosis, and rheumatoid arthritis (7-12). The α(v)β(3) integrin is strongly expressed on tumor cells and activated endothelial cells. In contrast, expression of the α(v)β(3) integrin is weak on resting endothelial cells and on most normal tissues. The α(v)β(3) antagonists are being studied as anti-tumor and anti-angiogenic agents, and the agonists are being studied as angiogenic agents for coronary angiogenesis (11, 13, 14). A tripeptide sequence comprising Arg-Gly-Asp (RGD) has been identified as a recognition motif used by extracellular matrix proteins (vitronectin, fibrinogen, laminin, and collagen) to bind to a variety of integrins, including α(v)β(3). Various radiolabeled cyclic RGD peptides have been introduced for tumor imaging and tumor angiogenesis (15). Ferritin is composed of 24 subunits of heavy and light chains that self-assemble to form a cage-like nanoparticle (nanocage) at physiological pH (7.4) with internal and external diameters of 8 nm and 12 nm (16, 17), respectively. The outer surface of ferritin can be chemically or genetically modified with ligands, and the cavity of ferritin can capture metal ions with high affinity (18). Lin et al. (19) genetically grafted an α(v)β(3)-targeted peptide (RDCRGDCFC, RGD4C) onto heavy chain ferritin (RGD4C-Fn) and chemically coupled Cy5.5 onto heavy chain ferritin (Cy5.5-Fn). Hybridization of RGD4C-Fn and Cy5.5-Fn (1:1) in the presence of (64)CuCl(2) resulted in the formation of RGD4C/Cy5.5-ferritin (64)Cu-loaded nanocages (RGD4C/Cy5.5-Fn-(64)Cu nanocages). RGD4C/Cy5.5-Fn-(64)Cu nanocages have been developed for positron emission tomography and NIRF multimodality imaging of tumor vasculature to study in vivo biodistribution of the tracer in tumor-bearing mice. RGD4C/Cy5.5-Fn-(64)Cu nanocages have been shown to have a high accumulation in the tumor vasculature and a predominant liver accumulation.
15504755 Plant alkaloid tetrandrine downregulates IkappaBalpha kinases-IkappaBalpha-NF-kappaB signa 2004 Dec Plant alkaloid tetrandrine (Tet), purified from Chinese herb Han-Fang Chi, is a potent immunomodulator used to treat rheumatic disorders, silicosis and hypertension in mainland China. We previously demonstrated that Tet effectively suppresses cytokine production and proliferation of CD28-costimulated T cells. In the present study, we investigated the possible involvement of nuclear factor kappa B (NF-kappaB) transcription factors, critical in CD28 costimulation, in Tet-mediated immunosuppression in human peripheral blood T cells. We showed that Tet inhibited NF-kappaB DNA-binding activities induced by various stimuli, including CD28 costimulation. At equal molar concentrations, Tet was as strong as methotrexate in suppressing CD28-costimulated NF-kappaB activities. Since Tet itself did not affect NF-kappaB binding to its corresponding DNA sequence, the results suggested that Tet might regulate NF-kappaB upstream signaling molecules. Further studies demonstrated that Tet could prevent the degradation of IkappaBalpha and inhibit nuclear translocation of p65 by blocking IkappaBalpha kinases alpha and beta activities. In addition, the activation of mitogen-activated protein kinases such as c-jun N-terminal kinase, p38 and extracellular signal-regulated kinase and activator protein-1 DNA-binding activity were all downregulated by Tet. Transfection assays performed in purified human peripheral blood T cells also confirmed the inhibition of NF-kappaB transcriptional activity by Tet. When four Tet analogues were readily compared, dauricine appeared to preserve the most potent inhibition on CD28-costimulated but not on H(2)O(2)-induced NF-kappaB DNA-binding activities. Our results provide the molecular basis of immunomodulation of Tet for being a potential disease-modifying antirheumatic drug in the therapy of autoimmune disorders like rheumatoid arthritis.
15356118 Stimulatory killer Ig-like receptors modulate T cell activation through DAP12-dependent an 2004 Sep 15 Stimulatory killer Ig-like receptors (KIRs) are expressed by various lymphocytes, including NK cells and subsets of T cells. In NK cells, KIRs associate with the adapter molecule KARAP/DAP12, which confers the ability to function as an independent activation unit. The function of KIRs and killer cell activating receptor-associated protein (KARAP)/DAP12 in T cells is unclear. By flow cytometry, we demonstrated that CD4+CD28null T cells heterogeneously express KIRs and/or KARAP/DAP12. In clones that lacked expression of KARAP/DAP12, the stimulatory KIR KIR2DS2 signaled through the JNK pathway, but did not activate the ERK pathway. However, in the presence of KARAP/DAP12, stimulation through KIR2DS2 led to phosphorylation of both JNK and ERK. Transfection experiments confirmed that KIR2DS2-mediated ERK phosphorylation was dependent on KARAP/DAP12. The differential signaling of KIR2DS2 through association with alternative adapter molecules resulted in differential regulation of cellular activity. In clones that lacked expression of KARAP/DAP12, stimulation of KIR2DS2 did not induce cytotoxicity. However, KIR2DS2 did augment suboptimal TCR stimulation, leading to enhanced IFN-gamma production. In clones that expressed KARAP/DAP12, KIR2DS2 directly activated both cytotoxicity and IFN-gamma production without the need for TCR-derived signals. The function of stimulatory KIRs in T cells is determined by the expression of the appropriate adapter molecule. Expression of KARAP/DAP12 is sufficient to convert a costimulatory KIR into a stimulatory molecule. These differing functions mediated by alternative signaling pathways have implications for the pathogenesis of diseases such as rheumatoid arthritis and acute coronary syndromes, in which aberrant expression of KIRs on T cells is frequently observed.
15203037 Liquid chromatography analysis of N-(2-mercaptopropionyl)-glycine in biological samples by 2004 Aug 5 N-(2-Mercaptopropionyl)-glycine (MPG) is a synthetic aminothiol antioxidant that is used in the treatment of cystinuria, rheumatoid arthritis, liver and skin disorders. Recent studies have shown that MPG can function as a chelating, cardioprotecting and a radioprotecting agent. Several other studies have shown that it may also act as a free radical scavenger because of its thiol group. Thiol-containing compounds have been detected in biological samples by various analytical methods such as spectrophotometric and colorimetric methods. However, these methods require several milliliters of a sample, time-consuming procedures and complicated derivatization steps, as well as having high detection limits. The present study describes a rapid, sensitive and relatively simple method for detecting MPG in biological tissues by using reverse-phase HPLC. With ThioGlo 3 [3H-Naphto[2,1-b] pyran, 9-acetoxy-2-(4-(2,5-dihydro-2,5-dioxo-1H-pyrrol-1-yl) phenyl-3-oxo-)] as the reagent, highly fluorescent derivatives of thiols can be obtained that are suitable for HPLC. MPG is derivatized with ThioGlo 3 and is then detected flourimetrically by reverse phase HPLC using a C18 column as the stationary phase. Acetonitrile: Water (75:25) with acetic acid and phosphoric acid (1 mL/L) is used as the mobile phase (excitation wavelength, 365 nm; emission wavelength, 445 nm). The calibration curve for MPG is linear over a range of 10-2500 nM (r=0.999) and the coefficients of the variation of within-run and between-run precision were found to be 0.3 and 2.1%, respectively. The detection limit was 5.07 nM per 20 microL injection volume. Quantitative relative recovery of MPG in the biological samples (plasma, lung, liver, kidney and brain) ranged from 90+/-5.3 to 106.7+/-9.3 %. Based on these results, we have concluded that this method is suitable for determining MPG in biological samples.
14573775 Expression of the angiogenic factor thymidine phosphorylase in THP-1 monocytes: induction 2003 Nov The angiogenic factor thymidine phosphorylase (TP) is highly expressed in human monocytes and macrophages, and its expression has been linked to the pathology and progression of solid tumors, rheumatoid arthritis, and gastric ulcers. In this study, TP mRNA and enzyme activity were found to be up-regulated upon the induction of differentiation of the human monocyte cell line THP-1 by phorbol 12-myristate 13-acetate (PMA). TP expression in THP-1 cells was similarly increased by tumor necrosis factor-alpha (TNFalpha). Because monocytes and macrophages are a predominant source of TNFalpha, the up-regulation of TP upon THP-1 differentiation could have been caused by the autocrine production of TNFalpha. In support of this hypothesis, PMA increased TNFalpha mRNA levels; furthermore, the increase in TP expression with PMA treatment was partially blocked by a neutralizing antibody to TNFalpha, particularly at the earlier time points. This data also suggested there may be additional mechanisms regulating TP expression upon PMA treatment of the cells. The induction of TP by TNFalpha was mimicked by an antibody to the TNFalpha receptor R2 (TNF-R2; p75), but not by an antibody to TNF-R1 (p55), suggesting that the TNF-R2 plays a role in the regulation of TP expression. The PMA-induced increase in TP expression was blocked by aspirin but not by the related agent indomethacin, suggesting that aspirin's effect was not caused by the inhibition of cellular cyclooxygenases. An alternative mechanism by which aspirin inhibits gene expression is the modulation of the transcription factor NFkappaB, and the TNFalpha-induced increase in TP mRNA was blocked by a cell-permeable NFkappaB inhibitory peptide. Furthermore, TNFalpha increased and aspirin (but not indomethacin) decreased NFkappaB DNA-binding activity in THP-1 cells. In conclusion, the modulation of TP expression in monocytes by pro- and anti-inflammatory agents suggests that its angiogenic-related actions could contribute to the inflammatory response associated with a number of pathophysiological conditions.
12750354 Differential regulation of VLA-2 expression on Th1 and Th2 cells: a novel marker for the c 2003 Jun We found that T(h)1 cells derived from ovalbumin (OVA)-specific TCR transgenic (DO11.10) mice showed significantly higher levels of VLA-2 (CD49b/CD29) expression than T(h)2 cells. In the early days (until 6 days) during induction of T(h)1 or T(h)2 cells, the expression of VLA-2 was gradually increased on both T(h) subsets. Thereafter, VLA-2 expression was further up-regulated on T(h)1 cells until 13 days, while a significant decrease of VLA-2 was observed in T(h)2 cells, resulting in a marked difference of expression at day 13. Up-regulation of VLA-2 on T(h)1 cells was not impaired in IFN-gamma(-/-) T(h) cells nor blocked by anti-IL-12 mAb treatment on wild-type T(h) cells, suggesting that up-regulation of VLA-2 on T(h)1 cells occurs in an IFN-gamma- and IL-12-independent manner. In contrast, T(h) cells cultured under IL-4-depleted T(h)2 conditions abrogated the down-regulation of VLA-2 expression, suggesting that down-regulation of VLA-2 expression on T(h)2 cells was dependent on IL-4. The finding that STAT6(-/-) T(h)2 cells did not show any down-regulation of VLA-2 expression and expressed the same levels of VLA-2 as T(h)1 cells indicated a critical role for the IL-4 receptor/STAT6 signaling pathway in IL-4-dependent down-regulation of VLA-2 on T(h)2 cells. Stimulation of T(h)1 cells by VLA-2 ligands such as collagen type I or agonistic mAb provided co-stimulation for anti-CD3 mAb-induced IFN-gamma production. However, these ligations had little effect on the IL-4 production of T(h)2 cells. Together, these results indicate that VLA-2 is a novel functional marker that dissociates T(h)1 from T(h)2 cells, and thus might be useful for therapeutic monitoring of T(h)1-dependent immune diseases such as rheumatoid arthritis or Crohn's disease.
12160520 Regulation of the interleukin-1 receptor antagonist in THP-1 cells by ligands of the perox 2002 Jun 21 Monocytes/macrophages (Mphi) play a pivotal role in the persistence of chronic inflammation and local tissue destruction in diseases such as rheumatoid arthritis and atherosclerosis. The production by Mphi of cytokines, chemokines, metalloproteinases and their inhibitors is an essential component in this process, which is tightly regulated by multiple factors. The peroxisome proliferator-activated receptors (PPARs) were shown to be involved in modulating inflammation. PPARgamma is activated by a wide variety of ligands such as fatty acids, the anti-diabetic thiazolidinediones (TZDs), and also by certain prostaglandins of which 15-deoxy-Delta(12,14)-PGJ2 (PGJ2). High concentrations of PPARgamma ligands were shown to have anti-inflammatory activities by inhibiting the secretion of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNFalpha) by stimulated monocytes. The aim of this study was to determine whether PGJ2 and TZDs would also exert an immunomodulatory action through the up-regulation of anti-inflammatory cytokines such as the IL-1 receptor antagonist (IL-1Ra). THP-1 monocytic cells were stimulated with PMA, thereby enhancing the secretion of IL-1, IL-6, TNFalpha, IL-1Ra and metalloproteinases. Addition of PGJ2 had an inhibitory effect on IL-1, IL-6 and TNFalpha secretion, while increasing IL-1Ra production. In contrast, the bona fide PPARgamma ligands (TZDs; rosiglitazone, pioglitazone and troglitazone) barely inhibited proinflammatory cytokines, but strongly enhanced the production of IL-1Ra from PMA-stimulated THP-1 cells. Unstimulated cells did not respond to TZDs in terms of IL-1Ra production, suggesting that in order to be effective, PPAR ligands depend on PMA signalling. Basal levels of PPARgamma are barely detectable in unstimulated THP-1 cells, while stimulation with PMA up-regulates its expression, suggesting that higher levels of PPARgamma expression are necessary for receptor ligand effects to occur. In conclusion, we demonstrate for the first time that TZDs may exert an anti-inflammatory activity by inducing the production of the IL-1Ra.
12006087 Selenium deficiency increases the expression of inducible nitric oxide synthase in RAW 264 2002 Aug 15 The inducible isoform of nitric oxide synthase (iNOS) is implicated in atherosclerosis, malignancy, rheumatoid arthritis, tissue and reperfusion injuries. A key determinant of the pro-oxidant versus protective effects of NO is the underlying redox status of the tissue. Selenoproteins, such as glutathione peroxidases (GPxs) and thioredoxin reductases, are key components of cellular defence and promote optimal antioxidant/oxidant balance. In this study, we have investigated the relationship between Se status, iNOS expression and NO production in Se-deficient and Se-supplemented RAW 264.7 macrophage cell lines. The cellular GPx activity, a measure of Se status, was 17-fold lower in Se-deficient RAW 264.7 cells and the total cellular oxidative tone, as assessed by flow cytometry with 2',7'-dichlorodihydrofluorescein diacetate, was higher in the Se-deficient cells than the Se-supplemented cells. Upon lipopolysaccharide (LPS) stimulation of these cells in culture, we found significantly higher iNOS transcript and protein expression levels with an increase in NO production in Se-deficient RAW 264.7 cells than the Se-supplemented cells. Electrophoretic mobility-shift assays, nuclear factor-kappaB (NF-kappaB)-luciferase reporter assays and Western blot analyses indicate that the increased expression of iNOS in Se deficiency could be due to an increased activation and consequent nuclear localization of the redox-sensitive transcription factor NF-kappaB. These results suggest an inverse relationship between cellular Se status and iNOS expression in LPS-stimulated RAW 264.7 cells and provide evidence for the beneficial effects of dietary Se supplementation in the prevention and/or treatment of oxidative-stress-mediated inflammatory diseases.
11978490 Nuclear factor-kappaB mediates over-expression of cyclooxygenase-2 during activation of RA 2002 May 1 Selenium (Se) is an essential micronutrient for all mammalian species and is associated with a variety of physiological functions, notably immune system, in the form of selenoproteins. Inadequate Se nutrition has been linked to various diseases, including rheumatoid arthritis, cardiomyopathy, and cancer. Important to this discussion is that cyclooxygenase-2 (COX-2) is over-expressed in all the aforesaid pathologies; however, a casual relationship between Se status and COX-2 expression remains to be established. The present study is based on the hypothesis that oxidant stress, a consequence of Se deficiency, lowers the activation potential of the redox-sensitive transcription factor, NF-kappaB, and that the activated NF-kappaB is required for the altered expression of COX-2. To test this hypothesis, we have investigated the relationship between Se status and COX-2 expression in response to LPS stimulation in RAW 264.7, a macrophage-like cell line. In Se-deficient cells, the Se-dependent glutathione peroxidase activity (Se-GPx), a measure of Se status, was markedly reduced and the overall oxidative stress was significantly higher than Se-supplemented cells. Upon lipopolysaccharide (LPS) stimulation, we found 2-3-folds higher COX-2 protein expression as well as higher PGE2 levels in Se-deficient cells than Se-supplemented cells. In comparison, COX-1 protein expression was not affected by either LPS stimulation or Se status. Following LPS stimulation, the nuclear localization of NF-kappaB was significantly increased in Se-deficient macrophages, thereby leading to increased expression of COX-2. This is the first report demonstrating an inverse relationship between Se status and the expression of COX-2.
15318291 [Evaluating the reliability, validity and responsiveness of the german short musculoskelet 2004 Aug BACKGROUND: Modern patient based outcome measures like the SMFA-D (German Short Musculoskeletal Function Assessment Questionnaire) are able to detect the impairment and functional capacity of patients with musculoskeletal extremity disorders. The SMFA-D was successfully evaluated in several cohorts treated operatively for osteoarthritis of the knee and hip, rotator cuff tears and rheumatoid arthritis. The aim of the present study was the evaluation of the SMFA-D in patients with conservative treatment for hip osteoarthritis. PATIENTS AND METHODS: 69 patients with osteoarthritis of the hip were enrolled in a prospective controlled clinical trial. All patients completed the SMFA-D, SF-36, WOMAC, FFbH-OA. A standardized test of walking speed and the functional status of the patient as judged by the physician were recorded. Statistical analysis were done for the following: re-test reliability (ICC), internal consistency (Cronbach's alpha), validity and responsiveness. RESULTS: Internal consistency (Cronbach's alpha) was alpha = 0.89 and alpha = 0.97 for the SMFA-D scales. The retest reliability (ICC, unjust, mixed effect) was 0.91 (p < 0.001) for the function index and 0.73 (p < 0.001) for the bother index. Both indices correlated significantly with the FFbH-OA (r = 0.66 to r = 0.84), the WOMAC (r = 0.55 to r = 0.86) and the scales of the SF-36 (r = - 0.34 to r = - 0.85) on all three time points, which supports construct validity. There was mainly a significant correlation between the SMFA-D scales and the functional status of the patient (r = 0.21 to r = 0.44), pain reported by the patient (r = 0.43 to = 0.54) and the self selected walking speed (r = 0.28 to r = 0.51), which supports external validity. We were able to differentiate operatively and conservatively treated patients (discriminant construct validity). At the end of the rehabilitation program we were able to demonstrate small to medium treatment effects in SMFA-D and SF-36. The WOMAC and FFbH-OA were not able to demonstrate these treatment effects. CONCLUSION: Even in patients with conservative treatment of hip osteoarthritis the SMFA-D represents a reliable, valid and responsive measure. The use of the SMFA-D can be recommended as a patient based outcome measure.