J. Lenting and K. Mertens, unpublished observations). Given the observation that VWF prevents binding of the FVIII procofactor to many of the FVIII receptors, it is possible that increased FVIII expression levels in the presence of VWF could partially be explained by VWF preventing re-uptake of FVIII by the producing cell. Provided that FVIII

escapes re-uptake by the cell, it is quickly captured into a complex with its carrier VWF [80]. VWF is crucial in maintaining FVIII plasma levels, which is illustrated by the severely reduced FVIII levels in patients that lack circulating VWF (e.g. von Willebrand disease type 3). The notion that FVIII levels are also reduced in cases of impaired complex formation (e.g. von Willebrand disease MAPK inhibitor type 2N) demonstrates that VWF protection is only valid when VWF and FVIII are circulating in the complex. Within this complex, VWF may prolong survival of FVIII in the circulation by preventing the interaction of FVIII with its clearance receptors. It is of importance to realize that this protection is not absolute! First, complex formation between FVIII and VWF follows the laws of thermodynamics in that the amount of FVIII that is in the complex is determined by the affinity constant for complex formation and the

concentrations of the individual proteins. Indeed, it has been reported that while the majority Cell Cycle inhibitor of FVIII (92–95%) is in complex with VWF, a small but significant portion (2–5%) circulates as free FVIII protein [81,82]. This portion is therefore susceptible to clearance by the various receptors. Second, the possibility exists that conditions at the cellular surface influence the stability of the FVIII/VWF complex, favouring its dissociation. For instance, Sarafanov et al. [70] proposed that the VWF/FVIII complex is NADPH-cytochrome-c2 reductase bound to HSPG at the cellular surface, resulting

in dissociation of the complex. Subsequently, FVIII is transferred to LRP1, whereas VWF is released back into the circulation. Meijer et al. [83] have recently presented a similar concept, based on studies using fluorescent FVIII-fusion proteins. Their observations differed from those by Sarafanov et al. [70] in that the complex only dissociated at the cellular surface following conformational changes within the VWF molecule. Taken together, the influence of VWF on FVIII clearance is apparently more complex and less straightforward than previously anticipated. What adds to this complexity is that FVIII may also be subject to clearance as part of the VWF complex. The multimeric VWF protein is subject to clearance as well, and it seems conceivable that a substantial part of FVIII is cleared while being bound to VWF. In support of this possibility is our observation that VWF and FVIII co-localized into similar cells, when injected as a complex into VWF-deficient mice [84]. This may seem in contradiction with some observations that VWF interferes with FVIII internalization by a number of cell types [32,33,83].

In addition, evaluating only a single late timepoint with reduced injury and fewer neutrophils is insufficient, as the decrease in neutrophil infiltration could simply be a consequence of less liver injury. Taken together, the present study, Selleckchem Small molecule library which in part repeats previous experiments and mistakes, does not support the hypothesis that neutrophils are critical for APAP hepatotoxicity. Hartmut Jaeschke Ph.D.*, Mitchell R. McGill B.Sc.*, C.

David Williams B.Sc.*, * Department of Pharmacology, Toxicology & Therapeutics, University of Kansas Medical Center, Kansas City, KS. “
“Pyogenic liver abscess is an uncommon disease in most countries. Approximately 60% of patients develop an abscess because of cholangitis associated with biliary and pancreatic disorders. Another 10% of patients develop liver abscesses because of intra-abdominal infections that spread to the liver, presumably via the portal vein. However, in at least 30% of patients, the cause of the abscess remains unclear (cryptogenic). In this latter group, colonoscopy has revealed a number of colonic disorders including colon cancer, benign tumors, multiple colonic ulcers and diverticulitis. A

group of particular interest 3-MA molecular weight is diabetic patients with abscesses caused by Klebsiella pneumoniae who have a relatively high frequency of colon cancer and advanced colonic polyps. Associations of pyogenic liver abscesses with gastrointestinal disorders outside the colon appear to be

infrequent. However, in this report, we describe an association of liver abscess with an ulcerated duodenal mafosfamide lipoma. A woman, aged 60 years, was treated with antibiotics at a local hospital because of a liver abscess. Although her symptoms had improved, she remained anemic and was referred for further evaluation. She was known to have hypertension and hyperlipidemia. Blood tests revealed anemia (hemoglobin 9.3 g/dl, 93 g/l) but her white blood cell count and C-reactive protein level had returned to the reference range. An abdominal computed tomography (CT) scan showed an abscess, 6 cm in diameter, in the right lobe of the liver. A positron emission tomography (PET)-CT scan revealed increased uptake in the third part of the duodenum as well as increased uptake in the right lobe of the liver (Figure 1). A decision was made to avoid percutaneous drainage of the liver abscess. Double-balloon enteroscopy was performed and showed a large pedunculated tumor, 5 × 2 cm in size, originating in the third part of the duodenum. The mucosa covering the lesion was normal but there was an area of ulceration at the apex. The duodenal lesion was thought to be a lipoma and was treated by endoscopic submucosal dissection rather than endoscopic mucosal resection. Histology confirmed the presence of a lipoma with apical ulceration and foci of suppuration (low and higher power, Figure 2).

However, the long term implications are unknown. Current guidelines suggest cessation of treatment in the last trimester of pregnancy to reduce fetal exposure but this is difficult for women with IBD who are not in deep remission, as active disease is a greater risk for adverse pregnancy outcome. This study aimed to examine drug levels of ATA in cord blood of newborns

exposed to ATA in pregnancy, and to correlate these with maternal levels, the duration of therapy during pregnancy, and time to clearance of ATA in infants. Methods: Women with IBD exposed to infliximab (IFX) or adalimumab (ADA) during pregnancy were included from 2012-present at 14 hospitals in Australia, New Zealand and Denmark. ATA levels were measured using an ELISA in cord and maternal blood at delivery (Matriks Biotek). If positive at birth, the infants were Abiraterone nmr tested every third month until ATA were undetectable. Demographics, disease phenotype, disease activity in pregnancy, duration of ATA use in pregnancy, medication and pregnancy outcomes were prospectively selleck products collected by questionnaire and from the treating doctor. Results: 80 pregnant women have been enrolled, and so far 53 mother-baby pairs have been tested (27 IFX and 26 ADA). An inverse correlation between duration since last exposure and cord ATA levels

at birth was found (IFX: r = −0.58, p = 0.002; ADA: r = −0.42, p = 0.047). This was also the case for maternal levels at birth (IFX: r = −0.59, p = 0.002; ADA: r = −0.52, p = 0.01). There was a strong NADPH-cytochrome-c2 reductase correlation between cord blood and maternal levels at delivery (IFX: Pearson’s r = 0.80, p < 0.0001; ADA: r = 0.80, p < 0.0001). Drug was ceased prior to gestational week (GW) 30 in 15 (28%) women. In them, mean serum concentrations were 0.81 μg/ml (IFX) and 0.08 μg/ml (ADA), and the cord blood level at delivery was <3 μg/ml in 11/15 (73%). So far 30 babies have completed testing for detectable

ATA levels, and testing is ongoing in the remaining 23 babies. Complete clearance of ATA was seen in 7, 5, 12 and 6 babies at birth, by 3, 6 and 9 months, respectively. To date there has been one detectable ATA level at 9 months. Three women (5.7%) gave birth preterm (GW 33–35). No congenital malformations were detected and all babies are developing normally. Conclusion: Maternal and neonatal ATA levels were inversely correlated with the duration since last exposure. Cord blood ATA levels were strongly correlated with maternal level at delivery. Maternal cessation of ATA prior to week 30 successfully reduced fetal exposure to drug in the vast majority of cases. Follow up will determine whether high neonatal levels have any negative consequences.

However, in addition to side effects common to immunomodulatory therapy, FTY720 was reported to cause cardiovascular complications, macular edema, and brain inflammation,4 presumably the result of interactions with more than one S1P-receptor subtype.8 Previously, we demonstrated that FTY720 induces www.selleckchem.com/products/Deforolimus.html apoptosis in HCC cells through the reactive oxygen species (ROS)-dependent activation of protein kinase C (PKC)δ.7 Dissociation of the apoptosis-inducing activity of FTY720 from its S1P receptor agonist activity provides a basis for its pharmacological

exploitation to develop a novel class of antitumor agents. Here, we report the development of a nonimmunosuppressive FTY720 analogue, OSU-2S [(S)-2-amino-2-(4-[(6-methylheptyl)-oxy]phenethyl)pentan-1-ol], which exhibits higher in vitro and in vivo potency than FTY720 in suppressing HCC cell

growth through PKCδ signaling. CA-Akt, constitutively active Akt; DAPI, 4,6-diamidino-2-phenylindole; DCFDA (5-(and-6)-carboxy-2′,7′-dichlorodihydrofluorescein diacetate); DMEM, Dulbecco’s modified Eagle medium; DMS, N,N-dimethylsphingosine; I-BET-762 concentration DPI, diphenyleneiodonium; FBS, fetal bovine serum; GST-π, glutathione S-transferase-π; HA, hemagglutinin; HCC, hepatocellular carcinoma; Hep3B-luc, luciferase-expressing Hep3B; i.p., intraperitoneal; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; PARP, poly (ADP-ribose) polymerase; p-FTY720, phosphorylated FTY720; PKCδ, protein kinase Cδ; PP2A, protein phosphatase 2A; ROS, reactive oxygen species; S1P, sphingosine-1-phosphate; siRNA, small interfering RNA; shRNA, short hairpin RNA; SphK2, sphingosine kinase 2; TLC, thin-layer chromatography; TMA, tissue Clomifene microarray. Details about reagents, their commercial sources, and experimental procedures are provided in the Supporting Information. The HCC cell lines, Hep3B, PLC5 and Huh7, and primary nonmalignant human hepatocytes were used in this study. FTY720 was synthesized as described,9 and p-FTY720 was purchased from Cayman Chemical (Ann Arbor, MI). Synthesis of OSU-2S and phosphorylated OSU-2S (p-OSU-2S) will be described elsewhere. Various polyclonal and monoclonal antibodies were used for western

blotting, immunocytochemical, and flow cytometric analyses. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assays as previously reported.10 For assessment of apoptosis, treated cells were stained with Annexin V-Alexa Fluor 488 and propidium iodide according to the vendor’s protocols. For caspase-3 activity, cells were incubated with the fluorogenic caspase-3 substrate (Ac-DMQD)2-Rh110 for 20 minutes. ROS production was detected using the fluorescence probe 5-(and-6)-carboxy-2′,7′-dichlorodihydrofluorescein diacetate (DCFDA) as described.7 Data were analyzed by ModFitLT V3.0 software program. Immunoblotting of biomarkers in cell lysates and tumor tissue homogenates was performed as reported.

In all seasons, L. guanicoe

occurrence was influenced by both environment and livestock interactions, especially small livestock (goats and sheep). Guanacos selected for habitats characterized by high temporal variability in plant productivity and away from potential human contact. In all seasons, L. guanicoe was negatively related to the RSPF of small livestock, but the reverse was not the case, suggesting that L. guanicoe avoids sites used by goats and sheep. In contrast, livestock was mainly affected by environmental variables related to human presence and was not affected by the interactions with herbivores. Contrary to our predictions, selleck chemicals goats and sheep were also associated with less productive sites, probably indicating strong degradation of

the sites to which they are restricted. Our results suggest a spatial segregation between L. guanicoe and domestic herbivores throughout the year, which is explained by competitive interactions of L. guanicoe with small livestock but also in response to vegetation productivity and human pressure. This study shows the importance of including species this website interaction effects in habitat modeling. “
“Ecological theory predicts that sympatric species should avoid competition through diet, spatial and/or temporal partitioning. In carnivores, interference is widespread between species with similar diets. Smaller species are expected to differentiate their diet from that of larger, dominant ones, to reduce the risk of potentially lethal encounters. Interference has been reported between tigers and common leopards, with the former dominant over the latter. In 2009–2011, in an area

of Terai, Branched chain aminotransferase South-West Nepal, we assessed food habits and prey selection of tigers and common leopards, to evaluate whether prey partitioning occurred between these large cats. Prey availability was high, both in terms of number of species (at least seven wild ungulates beside livestock, two primates and an array of smaller prey) and density (large ungulates, livestock and primates: 130.8–174.8 individuals per km2). Wild vertebrates were the staple of both cats (tigers: 82.7%; common leopards: 66.6%), but common leopards used livestock significantly more than tigers did. Diet breadth of leopards was c. 20% larger than that of tigers, indicating a broader trophic niche. Significant differences in prey use and selection occurred between tigers and leopards, with the former using large (i.e. >100 kg) prey more often and small (i.e. 5–25 kg) prey less often than the latter did. Medium-sized prey were taken in comparable proportions by the two cats, with a great overlap of diet (Pianka index: 0.85). In conclusion, in our study area, apparently tigers and leopards did not base their coexistence on diet partitioning, suggesting a major role for spatial and/or temporal partitioning.

Recently, vitamin D and its analogs have been deemed as potential regimen to treat a variety of cancers alone or in combination with other drugs. Although, the epidemiologic evidence regarding the association of vitamin D and hepatocellular carcinoma (HCC) is still inconclusive, biochemical evidence clearly indicates that HCC cells are responsive to the inhibitory effect of vitamin D and its analogs.

In this review, we discuss the current status of HCC and its treatment, the source, metabolism, functions, and the mechanism of actions of vitamin D, and the biochemical studies of vitamin https://www.selleckchem.com/products/abt-199.html D analogs and their implications in the prevention and treatment of HCC. Hepatocellular carcinoma (HCC), originating from epithelium of hepatocytes and accounting for 80% of primary liver cancers, ranks as 4th place in causing tumor-related deaths globally.1 HCC affects more U0126 cost than half a million people annually and the comparable incidence to its mortality rate demonstrates its dismal prognosis.1 About 80% of HCC is found in patients with cirrhotic liver2 with hepatitis B and C being the main causes of liver cirrhosis. The incidence of HCC in hepatitis B patients is 200 times as high as that of non-infected people and patients with hepatitis C have fivefold more chance to develop HCC than patients with hepatitis B.3 Other cases of non-viral related liver cirrhosis have also been found to be positively associated

with HCC, such as nonalcoholic steatohepatitis, hemochromatosis, alcoholic liver disease, alpha-1 antitrypsin deficiency, and autoimmune hepatitis. Moreover, some environmental toxins, such as aflatoxin B1, are also reported to incite the development of HCC.4 Generally, men are more vulnerable to HCC than women; especially in some areas, such as Africa

and Southeast Asia, the ratio of male-to-female could Buspirone HCl reach 3.7.5 Presently, partial hepatectomy remains the standard treatment for patients with resectable HCC and without obvious liver cirrhosis. However, growing evidence has suggested that liver transplantation and radiofrequency ablation of the tumor could provide comparable benefit on survival as well, compared to partial hepatectomy, especially when the tumors are smaller than 3 cm.6 For example, in Child–Pugh class A patients with a single tumor, the 5-year-survial rate could be improved to 70% after these radical therapies as compared to 65% 3-year-survial without any treatments.2 On the other hand, the advanced HCC patients, who are unfit for receiving radical therapies and are poor respondents to traditional chemotherapy and radiotherapy, usually have a survival time of less than 6 months.7 Finally, most HCC patients (70–80%)7 are diagnosed at intermediate-advanced stage and there is no effective treatment available at the present time.2,8 Under these bleak conditions, developing a new therapeutic regimen against HCC has been a priority.

In vivo plasmacytoid DC depletion was accomplished using 120G8 (200 μg; Imgenex, San Diego, CA).26 In selected experiments, the novel immune-modulator VAG539 (30 mg/kg, Novartis, Basel, Switzerland) was used to partially inactivate DC.27 To deplete Gr1+ cells, RB6-8C5 was employed (150 μg/day; Monoclonal Antibody Core Facility, Sloan-Kettering Institute, New York, NY). For in vivo NK cell depletions, 100 μL of a 1:5 dilution of anti-asialo GM1 (Wako Chemical, Richmond, VA) was injected intraperitoneally 3 days prior to APAP treatment. In selected experiments, antibodies

directed against IFN-α (1 μg, F18, Sigma), TNF-α (200 μg, AB-410, R&D, Minneapolis, MN), IL-6 (200 μg, AB-406, R&D), or MCP-1 (50 μg, AB-479, R&D) were administered in vivo before APAP challenge. Changes in serum R788 clinical trial liver enzymes, including ALT, aspartate aminotransferase (AST), were determined using the Olympus AU400 Chemistry Analyzer (Center Valley, PA). In survival experiments, animals were euthanized when they were moribund and death was imminent. Animal procedures were approved by the New York University School of Medicine Animal Care and Use Committee. Liver DC were isolated as described.25 Briefly, immediate postmortem laparotomy was performed and the portal vein was buy ACP-196 cannulated and infused with 1% Collagenase IV (Sigma-Aldrich). Hepatectomy was then performed and livers mechanically minced

before incubation with Collagenase IV at 37° for 10 minutes. Low-speed (30g) centrifugation was performed Liothyronine Sodium to exclude the pelleted hepatocytes followed by high-speed (300g) centrifugation to isolate the hepatic nonparenchymal cells (NPCs). The NPC were then further enriched over a 40% Optiprep

(Sigma-Aldrich) density gradient. To purify the DC population, NPCs were incubated with 1 μg of anti-FcγRIII/II (2.4G2, Fc block; Monoclonal Antibody Core Facility, Sloan-Kettering Institute) per 106 cells, labeled with fluorescently conjugated anti-CD11c and anti-MHC II (both BD Biosciences, Franklin Lakes, NJ), and FACS sorted using a MoFlo cell sorter (Beckman Coulter, Fullerton, CA). Splenocytes were prepared by mechanical disruption and specific cellular subgroups were purified by FACS. For in vitro T-cell proliferation assays, peptide-pulsed DC (3 × 104) were added to CD8+OT-I TCR-transgenic T cells (1 × 105) specific for Ova257-264, or CD4+OT-II TCR-transgenic T cells specific for Ova323-339 in 96-well plates for 48-72 hours before pulsing with 3H-thymidine as described.25 DCs were loaded with the relevant Ova peptide (10 μg/mL; AnaSpec, San Jose, CA) for 90 minutes before co-culture with respective T cells. In selected experiments, VAF347 (5 mM, Novartis), a low-molecular-weight compound that binds the aryl hydrocarbon receptor was used to prevent DC induction of CD4+ T cells.28 Western blotting was performed as described.29 Livers were minced in PBS with Protease Inhibitor cocktail (Roche, Pleasanton, CA) and homogenized.

Abrao Ferreira – Grant/Research Support: ABBOTT, ROCHE, BMS, JANSSEN; Speaking and Teaching: ROCHE, BMS, JANSSEN Djamal Abdurakhmanov – Grant/Research Support: Roche; Speaking and Teaching: BMS, Jansenn, MSD, Novartis Giovanni B. Gaeta – Advisory Committees or Review Panels: Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche, Janssen, Merk, BMS, Novartis, Gilead, Roche Filip Beeldens – Employment:

Janssen Research and Development Wafae Iraqi – Employment: Janssen Ralph DeMasi – Management Position: Johnson and Johnson Andrew Hill – Consulting: Janssen Joerg M. Lauffer – Employment: Janssen; Stock Shareholder: Janssen Isabelle Lonjon-Domanec – Employment: Janssen Massimo Colombo – Advisory Committees or Review Panels: BRISTOL-MEYERS- SCHERING-PLOUGH, ROCHE, GIlEaD, Ja’nssen Cilag, Achillion; selleck kinase inhibitor Grant/Research Support: BRISTOL-MEYERS-SQUIBB,

ROCHE, GILEAD, BRISTOLMEYERS-SQUIBB, ROCHE, GILEAD; Speaking and Teaching: Glaxo Smith-Kline, BRISTOL-MEYERS-SQUIBB, SCHERING-PLOUGH, C646 concentration ROCHE, NOVARTIS, GILEAD, VERTEX, Glaxo Smith-Kline, BRISTOL-MEYERS-SQUIBB, SCHERING-PLOUGH, ROCHE, NOVARTIS, GILEAD, VERTEX The following people have nothing to disclose: Petr Urbanek, Christophe Moreno, Inmaculada Fernandez, Adrian Streinu-Cercel Hepatitis C virus (HCV) exists as a quasispecies (QS) of related genetic variants. QS are thought to be an important factor in the evasion of the host immune response and the maintenance of chronic infection. Furthermore, a number of studies have demonstrated associations between selleck antibody QS complexity and diversity in the hypervariable region 1(HVR1) and sustained viral response to treatment. Many of these studies have either been retrospective, focused on acute infection, or post transplant changes and most have used variable sampling intervals of many months if not years. We recruited and sampled

the HCV HVR1 QS in 20 chronically infected individual at fortnightly for a total of 16 weeks. We analysed QS diversity, complexity, and divergence for a per sample mean of 16 (12-24) HVR1 clones which had been created using nested PCR. QS change was visualized using both phyelogenetic trees and median joining networks. We examined the samples for evidence of selection at both HVR1 wide and codon level. Finally, we investigated for evidence of multiple subpopulations. We demonstrate statistically significant less QS diversity and complexity in HVR1 QS in patients with cirrhosis (p<0.01). A number of cirrhotic patients maintain a homogenous QS profile for the entire study period which contrasts with non cirrhotic patients where marked change is found.

In addition, the composition of serum-free, hormonally defined medium (HDM) for the different stages is shown, and it is the same composition established

in previous studies.9 Rigorous purification of parenchymal cells away from their native mesenchymal cell partners resulted in a loss of viability of the parenchymal cells (especially the stem cells GSK2118436 concentration and progenitors), as shown previously.2, 9 Cocultures of hHpSCs with different subpopulations of mesenchymal feeder cells elicited distinct biological responses. Those with angioblasts remained stem cells, and those with precursors to hepatic stellate cells and endothelia became hepatoblasts; this provided distinctive antigenic, biochemical, and ultrastructural features for both parenchymal and mesenchymal cell populations (Figs. 2 and 3). The hHpSC/angioblast partnership resulted in cells that were tightly bound to one another on their lateral borders through large numbers of tight junctions, desmosomes, and interdigitated microvilli. Efforts to disperse the angioblasts and hHpSCs into single cells were not successful with the customary enzymes (e.g., trypsin, chymotrypsin, dispase, and collagenases), and they resulted in a rapid loss of cell viability. Mechanical passaging, as used for human embryonic stem cells in culture, resulted Birinapant cell line in reasonably

efficient passaging of hHpSCs13 and was used for the studies reported here. The hHB/stellate cell/endothelial cell precursor partnership resulted in cells that were more loosely bound to one another, as evidenced by both light microscopy and ultrastructural analyses. Transmission electron microscopy (TEM) observations confirmed that hHBs were distinct from hHpSCs: there were striking increases in the number and size of the desmosomes and the intermediate filaments that terminated at the desmosomes

in the mesenchymal cells and in the appearance of bile canaliculi. In parallel with morphological changes, hHBs had an antigenic profile that overlapped Grape seed extract with that of hHpSCs but showed distinctions in expressing ICAM-1 (not NCAM) and AFP and P450-A7 (data not shown). The activation of angioblasts, which gave rise to hHpSTCs and endothelial cell precursors, was associated with dramatically elevated levels of CD146 (Fig. 3) and with elevated levels of ASMA and desmin (data not shown); this all correlated with the formation of cords of hHBs and committed progenitors from the colonies of hHpSCs. Later lineage stages of parenchymal cells were partnered with either endothelia (hepatocytes) or hepatic stellate cells, pericytes, and myofibroblasts (cholangiocytes). The data from cultures of these epithelial-mesenchymal partnerships are not shown except in summary form in Supporting Information Fig. 7, although we provide data on the identified paracrine signals from those stages of mesenchymal cells.

21, 36 The βA domain of this structure shows a break in helix α1 and lacks a tight hydrophobic packing between residues of the central region of helix α1 and the top of helix α7 and the end of the β6-α7 loop. These characteristics persist during the simulation of the TC and GRGDSP complexes. In stark contrast, the simulation of the TUDC-bound ectodomain reveals a T-junction formation between these two regions that was first observed in the structure of liganded αIIbβ3,20 and afterwards in computational studies of agonist-bound integrin ectodomains.21, 22 As a consequence an outward and downward movement of helix α7 results that resembles

that of a rod connecting Tanespimycin datasheet a piston (the region of the T-junction) to a crankshaft (the hybrid domain). This mechanical model has been proposed as a mechanism to change the interdomain βA/hybrid domain hinge angle, leading to an unbending of the integrin structure.37 According to current models, such an unbending is required for activation.32 The activating structural changes in the case of the TUDC-bound

ectodomain are only observed in the vicinity of the binding pocket and remain located within the SB203580 βA domain (Supporting Fig. 8). This finding is in line with the timescale of integrin activation in the absence of force (microseconds38 to seconds39), which is orders of magnitude larger than the simulation time of 200 ns investigated here. The limited simulation time may also explain why no separation of the β6-α7 loop from the β1-α1 loop is observed in either simulation (Supporting Fig. 8). Further support for the observed T-junction formation in the β1-subunit is provided by the fact that residues involved in it (Fig. 6) are highly conserved across the eight β-subunits (Leu165: 6x, Val 367: 4x) as are residues in the immediate neighborhood (Leu169:

8x, Val370: 8x, Ile371: 6x). In total, the MD simulations suggest that TUDC, when interacting with the head region of α5β1 integrin, introduces an allosteric conformational change dipyridamole in the β1 subunit that propagates to the interdomain βA/hybrid domain hinge, which should lead to integrin activation, in agreement with experiment. In contrast, TC and GRGDSP do not introduce such a conformational change and, hence, should not activate integrin, again in agreement with experiment. As these ligands do remain bound in the head region, they effectively block this region for an interaction with TUDC, which can explain the TC-mediated inhibition of TUDC-induced β1 integrin activation. As to a possible mechanism of TUDC-induced β1 integrin activation, TUDC forms a monodentate MIDAS coordination with its sulfonic acid moiety (Fig. 6C), and with a second oxygen of this moiety a charge-assisted hydrogen bond with the amide nitrogen of Asn244 (Supporting Table 1).