Some critical genes are located in these modules, including GNAO1, GNAZ, PLCB1, CDC25B, LAMC1, FOS, ETS1, and SHC1 (Fig. 2B), suggesting that these genes may have important roles in the pathogenesis of HCC. To further determine which genes among these 362 DEGs represent novel cancer genes, we next examined the potential driver roles of the candidates in hepatic carcinogenesis. By combining the results of interaction network analysis with functional suggestions obtained from PubMed data mining, a set of 20 potential genes in frequently aberrant regions (amplification ≥12 samples and deletion ≥4 samples, respectively) were chosen

for further functional assessments (Supporting Table 3). Specifically, Selleck ITF2357 eight genes (DDEF1, SHC1, HAX1, RAD21, MPZL1, YWHAZ, SERPINA5, and GNAO1) were selected according to the results of network analysis (Fig. 2B). Four genes selected (PEA15, MK-2206 cost ILF2, MT1G, and PER1) have been identified previously in HCC.20-23 Eight genes selected (SNRPE, C1ORF2, BOP1, HEY1, DUSP12, C8ORF4, SLC25A4, and TRIM35) have been reported in other tumors.24-31 First, the mRNA expression levels of these 20 genes were confirmed by

quantitative real-time polymerase chain reaction (q-PCR) analysis in 47 of the 49 paired HCC samples (Supporting Fig. 2A). The results indicated that there was greater than two-fold up-regulation of HEY1 in 42.6% of HCC tumors (20/47), whereas there was greater than two-fold down-regulation of TRIM35 in 60% of HCC tumors (27/47) compared with matched noncancerous tissues. Additionally, in order to choose suitable cell line tools for the following functional assessments, the expression levels of these 20 genes in eight liver cancer cell lines were determined by q-PCR PJ34 HCl (Supporting Fig. 3). To investigate whether these genes are involved in liver tumorigenesis,

they were overexpressed using a lentivirus vector in SMMC-7721 and Huh-7 cells, which was confirmed by q-PCR (Supporting Fig. 4). The results showed that of the seven deleted genes, TRIM35 was capable of significantly inhibiting the in vitro cell proliferation and in vivo tumor growth of both SMMC-7721 and Huh-7 cells, whereas of the 13 amplified genes, HEY1 and SNRPE were capable of significantly promoting the proliferation of both SMMC-7721 and Huh-7 cells in vitro and in vivo (Fig. 3A-D). Consistent with the above results, knockdown of TRIM35 was observed to markedly promote HCC cell proliferation, whereas knockdown of HEY1 and SNRPE significantly suppress HCC cell proliferation, based on small interfering RNA (siRNA) analyses in HepG2 cells (Fig. 3E). Taken together, wededuced that TRIM35 may be a new tumor suppressor candidate and that HEY1 and SNRPE may be novel putative oncogenes in HCC.

This study was undertaken to evaluate the correlation between the intercanthal width

and interalar width with intercanine distance, in North Indian male and female patients for predicting the mesiodistal width of the maxillary anterior Acalabrutinib cell line teeth during tooth selection. Materials and Methods: The study was conducted with 100 North Indian patients (50 men, 50 women) ranging in age from 17 to 21 years. A digital caliper with an accuracy of 0.01 mm was used to measure the intercanthal and interalar width. A T-shaped flat metal plate (canine tip marker) was used to mark the intercanine distance, which was then measured with the digital caliper. These measurements were interpreted and subjected to statistical analysis. Student’s t-test was applied

MG-132 purchase to test the correlation between intercanthal width and interalar width with intercanine distance. Results: Calculated t-values between intercanine distances with interalar width in both male and female groups were 3.14 and 3.56, respectively, greater than the standard value taken at a 5% level of significance with 48 degree of freedom, showing a higher correlation of interalar width with the intercanine distance. Values obtained between intercanthal width and intercanine distance were lower than the standard value in both groups. Conclusions: A significant correlation was found between interalar width and intercanine distance in both men and women, suggesting that interalar width can be used as a reliable guide for maxillary anterior teeth selection. “
“Purpose: The purpose of this article is to analyze data from the results of the 2008 Survey of Pro Bono Services Adenosine triphosphate Provided by Practicing Prosthodontists. Survey results are used to examine characteristics and to compare the charitable care rendered by practicing prosthodontists to the dental field at large. Materials and Methods: The character and incidence of pro bono services (PBS) provided by prosthodontists are based on a 2008 survey, made possible through an American College of Prosthodontists Board of Directors’ sponsored initiative. Survey results are used to assess

the distribution of respondents practicing the specialty of prosthodontics in the United States, percentage of prosthodontists who render pro bono dental services for the community, percentage of total patient care devoted to pro bono treatment at no charge, number of patients treated annually with PBS, monetary value of pro bono care annually, types of pro bono procedures, percentage of practitioners using Prosthodontic Diagnostic Index (PDI), PBS by PDI category to assess complexity of donated work, and percentage of practicing prosthodontists using informatics to track services by the PDI. Results: Thirty-nine states were represented in the survey data. The highest responses were in the most populous states. The percentage of practicing prosthodontists providing PBS was 71.7%.

16 Therefore, the present study was designed to develop a murine model of chronic-binge ethanol administration to induce significant liver injury and

to test the hypothesis that treatment of IL-22 ameliorates alcoholic liver injury by using this model. Our findings demonstrated that chronic feeding plus a single dose of ethanol delivered by oral gavage induced more severe forms of liver injury and fatty liver than chronic find more feeding or single ethanol gavage alone. By using this model, we also demonstrated that IL-22 treatment ameliorated alcoholic liver injury, suggesting therapeutic potential for IL-22 in treating alcoholic liver disease. IL, interleukin; PCR, polymerase chain reaction; STAT3, signal transducer and activator of transcription 3; STAT3Hep−/− mice, hepatocyte-specific STAT3 knockout mice; TNF-α, tumor necrosis factor α. C57BL/6N mice were purchased from the NCI (Frederick, MD). Hepatocyte-specific STAT3 knockout (STAT3Hep−/−) mice and wild-type mice were described previously.24 All male mice were used unless specified. buy STA-9090 All animal experiments were approved by the NIAAA Animal Care and Use Committee. Eight- to 10-week-old male C57BL/6N mice were fed a nutritionally

adequate liquid control diet (Bioserv, Frenchtown, NJ) for 5 days, then divided into two groups (Supporting Information Fig. 1A): ethanol groups were fed a liquid diet containing 5% ethanol for 10 days; control groups were pair-fed control diet for 10 days; At day 11, mice in ethanol groups were gavaged a single doses of ethanol (5 g/kg body weight, 20% ethanol), whereas mice in control groups were gavaged isocaloric dextrin maltose. The gavage was always performed in the early morning. After gavage, mice were kept on control or ethanol diet and kept in the cages on the warm blanket with circulating water. All mice survived after chronic-binge ethanol feeding. After gavage, mice were slow-moving, but Tyrosine-protein kinase BLK conscious and regained normal behavior within 4-6 hours. The mice were always euthanized 9 hours after gavage when the serum levels

of ALT and AST reached to the peak. In some experiments, mice were also euthanized at different time points after gavage as specified. Mice were fed ethanol diet for 10 days and then treated (intraperitoneally) with a single dose of recombinant murine IL-22 protein (1 μg/g; GenScript, Piscataway, NJ) or saline. After treatment with IL-22 for 3 hours, mice were gavaged with a single dose of ethanol (5 g/kg) or maltose and sacrificed 9 hours after gavage. IL-22 adenovirus was made by cloning mouse IL-22 cDNA (544 bp) into the pENTR/D-TOPO system (Invitrogen), followed by using Invitrogen Gateway system to perform a LR reaction with pAd/CMV/V5-DEST to make the expression vector pAd/CMV/mIL-22.

The diagnosis is based on the combination of biochemical, autoimmune, and histological parameters, and exclusion of other liver diseases. Standard therapy consists of a combination of corticosteroids and azathioprine, which is efficacious in 80% of patients. Alternative therapies

are increasingly being explored in patients who do not respond to the standard treatment and/or have unacceptable adverse effects. This review examines the role of alternative drugs (second-line agents) available for AIH treatment non-responders. These agents include budesonide, mycophenolate mofetil, cyclosporin, tacrolimus, 6-mercaptopurine, 6-thioguanine, rituximab, ursodeoxycholic acid, rapamycin, and methotrexate. In addition, the risk of opportunistic infections MK1775 and malignancies are discussed. A treatment algorithm selleck chemicals is proposed for the management of patients with AIH treatment non-responders. “
“Liver tumor-initiating cells (T-ICs) are capable of self-renewal and tumor initiation and are more chemoresistant to chemotherapeutic drugs. The current therapeutic strategies for targeting stem cell self-renewal pathways therefore represent rational approaches for cancer prevention

and treatment. In the present study, we found that Lup-20(29)-en-3β-ol (lupeol), a triterpene found in fruits and vegetables, inhibited the self-renewal ability of liver T-ICs present in both hepatocellular carcinoma (HCC) cell lines and clinical HCC samples, as reflected by hepatosphere formation. Furthermore, lupeol inhibited in vivo tumorigenicity in nude mice and down-regulated CD133 expression, which was previously shown to be a T-IC marker for HCC. In addition, lupeol sensitized HCC cells to chemotherapeutic agents through the phosphatase and tensin homolog (PTEN)–Akt–ABCG2 pathway. PTEN plays a crucial role in the self-renewal and chemoresistance of liver T-ICs; down-regulation of PTEN by a lentiviral-based approach reversed the effect of lupeol on liver T-ICs. Using an in vivo chemoresistant

HCC tumor model, lupeol dramatically decreased the tumor volumes of MHCC-LM3 HCC cell line-derived xenografts, and the effect was equivalent to that Amylase of combined cisplatin and doxorubicin treatment. Lupeol exerted a synergistic effect without any adverse effects on body weight when combined with chemotherapeutic drugs. Conclusion: Our results suggest that lupeol may be an effective dietary phytochemical that targets liver T-ICs. (HEPATOLOGY 2011.) Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world.1 The curative treatment for HCC is liver transplantation or surgical resection.2, 3 However, 80% of HCC cases are presented at advanced stages and are no longer operable. Even after surgical resection, the long-term prognosis of HCC remains unsatisfactory due to high recurrence rates. For HCC patients in advanced stages, chemotherapy by way of either transarterial chemoembolization or systemically is the second-line treatment.

The diagnosis is based on the combination of biochemical, autoimmune, and histological parameters, and exclusion of other liver diseases. Standard therapy consists of a combination of corticosteroids and azathioprine, which is efficacious in 80% of patients. Alternative therapies

are increasingly being explored in patients who do not respond to the standard treatment and/or have unacceptable adverse effects. This review examines the role of alternative drugs (second-line agents) available for AIH treatment non-responders. These agents include budesonide, mycophenolate mofetil, cyclosporin, tacrolimus, 6-mercaptopurine, 6-thioguanine, rituximab, ursodeoxycholic acid, rapamycin, and methotrexate. In addition, the risk of opportunistic infections selleck kinase inhibitor and malignancies are discussed. A treatment algorithm click here is proposed for the management of patients with AIH treatment non-responders. “
“Liver tumor-initiating cells (T-ICs) are capable of self-renewal and tumor initiation and are more chemoresistant to chemotherapeutic drugs. The current therapeutic strategies for targeting stem cell self-renewal pathways therefore represent rational approaches for cancer prevention

and treatment. In the present study, we found that Lup-20(29)-en-3β-ol (lupeol), a triterpene found in fruits and vegetables, inhibited the self-renewal ability of liver T-ICs present in both hepatocellular carcinoma (HCC) cell lines and clinical HCC samples, as reflected by hepatosphere formation. Furthermore, lupeol inhibited in vivo tumorigenicity in nude mice and down-regulated CD133 expression, which was previously shown to be a T-IC marker for HCC. In addition, lupeol sensitized HCC cells to chemotherapeutic agents through the phosphatase and tensin homolog (PTEN)–Akt–ABCG2 pathway. PTEN plays a crucial role in the self-renewal and chemoresistance of liver T-ICs; down-regulation of PTEN by a lentiviral-based approach reversed the effect of lupeol on liver T-ICs. Using an in vivo chemoresistant

HCC tumor model, lupeol dramatically decreased the tumor volumes of MHCC-LM3 HCC cell line-derived xenografts, and the effect was equivalent to that ifoxetine of combined cisplatin and doxorubicin treatment. Lupeol exerted a synergistic effect without any adverse effects on body weight when combined with chemotherapeutic drugs. Conclusion: Our results suggest that lupeol may be an effective dietary phytochemical that targets liver T-ICs. (HEPATOLOGY 2011.) Hepatocellular carcinoma (HCC) is the fifth most common cancer in the world.1 The curative treatment for HCC is liver transplantation or surgical resection.2, 3 However, 80% of HCC cases are presented at advanced stages and are no longer operable. Even after surgical resection, the long-term prognosis of HCC remains unsatisfactory due to high recurrence rates. For HCC patients in advanced stages, chemotherapy by way of either transarterial chemoembolization or systemically is the second-line treatment.

However, consistent amplicons were not obtained from every reaction of the second amplification. We have re-evaluated the preparation

of mRNA and the amplification of cDNA to elucidate why the same amplicon was not provided every time. However, we could not resolve the issue. This may reflect the low abundance of such a variant. Therefore, we evaluated the patient’s mRNA by the result of 10 independently performed reactions. Although the method mentioned above is an excellent method for analysing ectopic F8 mRNA, in the case of some splice variants it is suggested that careful evaluation and selection of analyses are necessary. Originally, the patient was identified as having very mild congenital haemophilia A. The patient had no history of haemorrhage that required treatment until the detection of low FVIII activity level at the age of 60, MLN0128 clinical trial although he had showed some difficulty of haemostasis, for example in tooth extractions etc. during childhood. The fact that there is agreement between both the FVIII levels at a preoperative examination and the F8 mRNA levels described in the present study INCB024360 order supported the classification of the patient as having mild congenital haemophilia. However, at the present time, the patient has fallen into a very severe state due to development of anti-FVIII antibody. The inhibitor development process of the patient was typical, and took less than 20 exposure days from the first

FVIII concentrate injection [18]. Generally, inhibitor development in congenital haemophilia is more frequently observed in the severe patients null mutations

[8]. else It is comparatively rare that a patient with mild haemophilia A should develop the inhibitor. Inhibitor development in mild haemophilia A is typically observed in patients with molecular abnormalities because endogenous abnormal mutant FVIII, a cross-reacting material (CRM), is recognized as “self” and exogenously infused normal FVIII molecule is recognized as “non-self”. The developed antibody is often seen to cross-react with not only “non-self” but also “self”. This patient was diagnosed with congenital mild haemophilia A and has CRM as previously stated. However, analysis of the mRNA might suggest that this patient’s CRM would be normal FVIII, produced by the normal mRNA which avoided abnormal splicing. Therefore, this is an interesting case because the inhibitor in this patient raises the possibility that the nature and developing mechanism are different from the inhibitor usually developed in congenital mild haemophilia A. The inhibitor showed a type I inhibition kinetic pattern [19], predominantly IgG4 subclass [20], and multi-clonal epitopes (A2 domain and the light chain of FVIII). These characteristics were most typical of an alloantibody developed in congenital haemophilia. Moreover, we investigated the genetic risk factors in consideration of the possibility that the patient’s antibody developed as an autoantibody.

On

average patients completed 11 weeks of therapy at the time of data analysis. None of the pts died while on therapy, one patient (5%) on ribavirin discontinued all treatment due to AKI and severe anemia and one patient who was listed Trichostatin A for chronic rejection received re-transplantation while on therapy and his HCV RNA remained negative after transplant. In the SOF+SMV group median (IQR) creatinine levels were significantly worse at week 8 of therapy compared to BL(1.00 (0.95-1.5) to 1.7 (1.05, 1.95)), p=0.026.There was a trend towards an increase in tacrolimus level in this group (4.5 (3.9, 4.85) at baseline to 5.5 (4.1 to 6.45) on week 4), p = 0.066. There was no decline in hemoglobin level in the SOF + SMV group at weeks 4 or 8. In the SOF + RBV group, there was no effect on creatinine or TAC levels. All patients had undetectable HCV RNA with normalization of ALT/AST at week 4(Median ALT 56 to 19 and AST 78 to 22, p=0.008). Conclusion: In our experience, SOF-based regimens

were safe and effective in treating HCV recurrence. The combination of SOF+SMV was associated with an increase in creatinine levels and a mild increase in TAC levels which requires further www.selleckchem.com/products/epacadostat-incb024360.html monitoring. Disclosures: John J. Fung – Advisory Committees or Review Panels: Astellas, Novartis; Consulting: Vital Therapies; Grant/Research Support: Sanofi Naim Alkhouri – Advisory Committees or Review Panels: Gilead

Sciences The following people have nothing to disclose: Mohammed Eyad Yaseen Alsab-bagh, Ibrahim A. Hanouneh, Binu V. John, John K. Guirguis, Bijan Eghtesad, Nizar N. Zein Background: Non-interferon based therapy for chronic hepatitis C genotype 1 is a global goal. To date, there is limited data on the use of an all-oral treatment regimen in the post-liver transplant population. Methods: Thirty seven patients transplanted for genotype 1 chronic HCV were treated using an off-label combination of sofosbuvir, simeprevir, +/− ribavirin for 12 weeks. We collected data on patient characteristics, viral response, laboratory Nitroxoline values, and adverse events. The decision to treat patients for recurrent HCV post-transplant was made by one of 6 transplant hepatologists. Patients were monitored with monthly clinic visits and lab testing every 2-4 weeks during treatment. Results: Twenty five patients had genotype 1a, 7 patients had genotype 1b, and 5 patients were undifferen-tiated. There were 27 males (73%) and 14 African Americans (38%). The average age was 57 years (range 32-69) and the average time from transplant was 4 years (range 4-334 months). Twenty patients (54%) were treatment-experienced. Eight patients (22%) had recurrent cirrhosis. Sixteen patients (43%) were treated with ribavirin;13 started ribavirin at onset of treatment and 3 had ribavirin added due to slow viral response.

On

average patients completed 11 weeks of therapy at the time of data analysis. None of the pts died while on therapy, one patient (5%) on ribavirin discontinued all treatment due to AKI and severe anemia and one patient who was listed PXD101 purchase for chronic rejection received re-transplantation while on therapy and his HCV RNA remained negative after transplant. In the SOF+SMV group median (IQR) creatinine levels were significantly worse at week 8 of therapy compared to BL(1.00 (0.95-1.5) to 1.7 (1.05, 1.95)), p=0.026.There was a trend towards an increase in tacrolimus level in this group (4.5 (3.9, 4.85) at baseline to 5.5 (4.1 to 6.45) on week 4), p = 0.066. There was no decline in hemoglobin level in the SOF + SMV group at weeks 4 or 8. In the SOF + RBV group, there was no effect on creatinine or TAC levels. All patients had undetectable HCV RNA with normalization of ALT/AST at week 4(Median ALT 56 to 19 and AST 78 to 22, p=0.008). Conclusion: In our experience, SOF-based regimens

were safe and effective in treating HCV recurrence. The combination of SOF+SMV was associated with an increase in creatinine levels and a mild increase in TAC levels which requires further RG7420 monitoring. Disclosures: John J. Fung – Advisory Committees or Review Panels: Astellas, Novartis; Consulting: Vital Therapies; Grant/Research Support: Sanofi Naim Alkhouri – Advisory Committees or Review Panels: Gilead

Sciences The following people have nothing to disclose: Mohammed Eyad Yaseen Alsab-bagh, Ibrahim A. Hanouneh, Binu V. John, John K. Guirguis, Bijan Eghtesad, Nizar N. Zein Background: Non-interferon based therapy for chronic hepatitis C genotype 1 is a global goal. To date, there is limited data on the use of an all-oral treatment regimen in the post-liver transplant population. Methods: Thirty seven patients transplanted for genotype 1 chronic HCV were treated using an off-label combination of sofosbuvir, simeprevir, +/− ribavirin for 12 weeks. We collected data on patient characteristics, viral response, laboratory next values, and adverse events. The decision to treat patients for recurrent HCV post-transplant was made by one of 6 transplant hepatologists. Patients were monitored with monthly clinic visits and lab testing every 2-4 weeks during treatment. Results: Twenty five patients had genotype 1a, 7 patients had genotype 1b, and 5 patients were undifferen-tiated. There were 27 males (73%) and 14 African Americans (38%). The average age was 57 years (range 32-69) and the average time from transplant was 4 years (range 4-334 months). Twenty patients (54%) were treatment-experienced. Eight patients (22%) had recurrent cirrhosis. Sixteen patients (43%) were treated with ribavirin;13 started ribavirin at onset of treatment and 3 had ribavirin added due to slow viral response.

Incident primary headache disorders are less common in the elderly but do occur, and primary headache disorders such as migraine may persist PD98059 price into late life. The treatment of headache disorders in the older patient can be complex. Medical comorbidities, polypharmacy, and derangements of drug metabolism and clearance often limit medication choices. In this chapter we first briefly address secondary headache disorders in this age group, and

then discuss primary headache disorders in the context of their epidemiology, clinical features, and management strategies. We outline a multifaceted approach, including the use of acute, transitional, and preventive medications, interventional and nonpharmacological techniques, and addressing risk factors for headache progression and perpetuation. “
“We encourage athletic activity as a means to healthier bodies and minds, but sometimes, despite the best intentions, a player receives a blow to the head or body resulting in ongoing headaches. It is estimated that about 90% of these mild injuries resolve with the athlete being symptom-free in a week. Unfortunately, 10% will be left with ongoing headaches and other neurological symptoms. Natural Product Library purchase What is concussion? It is an injury to the head that results in changing the way the brain

functions. Concussions can also occur when there is a fall or blow to the body causing a jolt such that the brain moves rapidly back and forth. Post-concussive symptoms are usually mild but can result in confusion, headache, memory loss, impaired concentration, altered judgment, imbalance, and lack of coordination. Other symptoms commonly experienced are dizziness, fatigue, irritability, anxiety, depression, and a change in sleep pattern. In order for a headache disorder to be considered as caused by a head injury, it should appear or worsen

within a week of the concussion. The most common type of headache after such an injury is migraine, with tension-type headache being a close second. Post-concussion Carbachol migraine will result in an athlete developing nausea or sensitivity to light and noise with headache. Tension-type headache is without these features. Early after a concussion, it is advised to avoid any activities that could result in a second concussion, particularly before an athlete has recovered from the first. Athletes should not go back to playing the game again if any symptoms persist. If, for example, it is difficult to remember events immediately before or after a concussion, or if thinking and memory remain dulled, it is time to sit out vigorous activity until thinking and memory return to normal. Computed tomography scans (CT scans) can be helpful in ruling out serious bleeding injuries, but they cannot diagnose a concussion. With concussion, it is believed that there is a change in brain metabolism that causes the cascade of symptoms, including inflammation and chemical changes that result in headaches.

Cryosections of liver tissue were stained with a monoclonal rat antimouse CD31 antibody (BD Biosciences) as well as with a monoclonal goat antihuman vWF antibody (C-20, Santa Cruz) followed by a suitable secondary

antibody conjugated with Alexa fluor 488 (Invitrogen) to highlight microvessels. Sections were counterstained for nuclei Navitoclax in vivo with Vectashield Mounting Medium with DAPI (Vector Laboratories). Microvessel density (MVD) was determined by counting the CD31 positive vessels in three high-power fields (×100 magnification) from areas of highest vascularization. 21 The vWF-positive cells were quantified using NIH ImageJ software. Immunohistochemical staining of the monoclonal rat antimouse CD34 antibody (BD Biosciences) was performed on deparaffinized tissue sections using a routine avidin-biotin-immunoperoxidase technique (Vectastain ABC kit, Vector Laboratories). Before incubation with the primary antibody, tissue sections were subjected to microwave treatment with Citrate-based Antigen Unmasking Solution Ivacaftor cost (Vector Laboratories), followed by a 20-minute cool-down and treatment with 3% hydrogen peroxide. Angiogenesis-related perfusion was assessed by two-dimensional SonoVue-enhanced ultrasound imaging using a clinical imaging system (Acuson S2000, Siemens Healthcare). A multi-D matrix array transducer was attached to a device and the acoustic focus was placed on the

level of the dorsal liver capsule. After tail vein injections of mice with 50 μL diluted SonoVue (Bracco; diluted 1:5 with 0.9% NaCl), imaging in CPS-mode with a rate of 13 frames/sec for 40 seconds was performed. A region of interest (ROI) was set within the liver and the area under the curve (AUC) was

quantified for this ROI. The values were normalized by the AUC of an ROI placed in the caval vein (Supporting Fig. 1). Fluorescence labeling of VEGFR2 antibody was performed with VivoTag-S680 (VisEn Medical) by way of NHS ester. Normal goat IgG antibody (AF644 and AB-108-C, R&D Systems) was used as isotype control. Antibodies were diluted in 200 μL carbonate/bicarbonate Glycogen branching enzyme buffer to concentrations of 1 mg/mL and incubated each with 6 μL VivoTag-S680 for 1 hour at room temperature. Nonreacted VivoTag-S680 fluorophores were separated from labeled antibodies by size exclusion chromatography with fast protein liquid chromatography (FPLC) (ÄKTApurifier 10, GE Healthcare). Subsequently, VivoTag-S680 labeled VEGFR2 and IgG probes were injected in CCl4-treated Cxcr3−/− and WT littermates by way of a tail vein catheter (100 μL). Probe enrichment in the liver was determined 6 hours after probe injection using fluorescence molecular tomography (FMT 2500; Visen). Additionally, low-dose μCT scans (TomoScope DUO, CT Imaging) were performed and coregistered to the 3D FMT data in order to add anatomical information. Isolation of total protein and RNA from snap-frozen liver tissue samples were performed as described.