Methods: Assays were established with cultured HuH-7 cells with IC50 doses of APAP for MTT cell viability

assays. DNA damage was analyzed by γH2AX, pATM and pCHEK2 stainings or westerns and Comets for double-strand breaks. Cell cycling used FACS including after cell synchronization in S by hydroxyurea. Expression of 60 cell cycle regulators was analyzed by phosphoprotein array. Selected changes were examined in human liver explants after OLT. Whether this restriction could be reversed was also examined in cultured cells. Results: APAP-treated HuH-7 cells showed rapid depletion over 4h of cells in G1/S and M; subsequently, cells were arrested in G0/ G1. APAP cytotoxicity was associated with early decrease in ATM expression Selleckchem IWR1 followed later by greater pATM, γH2AX, pCHEK2 expression and Comet formation. Replicative stress typical of this DNA damage setting was verified by rapid destruction by APAP of cells synchronized in late G1/S. Analysis of cell cycle phosphoproteins with categorization as de novo appearance or >2-fold up- or down-regulation identified dysregulation of multiple regulators (e.g., ATM, CDC25C, CDC34, CDC37, CDK7, CDK1, CDK3, CDK8, CUL1, CUL2, CUL3, CCNE, E2F2, GSK3b, Ki67, RBL2, P19, CDKN1C and CDKN1A). Pathway mapping confirmed these regulators are important

in DNA damage-associated restrictions in G0/ G1 via G1/S checkpoints or other mechanisms. These mechanisms were human-relevant since explants in APAP-induced ALF showed widespread hepatic MCE nuclear staining of CDKN1A. learn more Remarkably, treatment of HuH-7 cells with an activator of Jak-Stat signaling decreased ATM-associated DNA damage and abrogated cell cycle arrest after APAP toxicity. Conclusions: APAP hepatotoxicity produced replicative

stress and arrest of residual cells in G0/G1 due to dysregulated ATM signaling and DNA damage. This replicative state involved cell cycle checkpoint controls and was reversible. Therefore, therapeutic interventions to restore cell cycling will be helpful for treating APAP-induced ALF. Disclosures: The following people have nothing to disclose: Preeti Viswanathan, Sriram Bandi, Sanjeev Gupta Background: Liver inflammation drives liver fibrosis and marks the transition from reversible to advanced stages of chronic liver diseases. Although liver inflammation is required for liver fibrogenesis, it is not known whether other events, such as hepatocyte death, are required for the development of liver fibrosis. Both liver inflammation and hepatocyte death are controlled by the Interferon regulatory factor 3 (IRF3), an innate immune protein involved in production of inflammatory cytokines and type-I interferons (IFNs), and in hepatocyte apoptosis. In the liver, IRF3 is activated via the Stimulator of Interferon Genes (STING). Aim: To investigate whether hepatocyte death is an independent determinant of liver fibrogenesis.

1a). During the late dry season, bush savanna associated with finer-grained sandstone became most strongly favoured, along with increased use of C. mopane tree savanna on shale and mudstone, particularly in the wetter of the 2 years. Zebra strongly favoured the open bush savanna associated with basaltic soils, the most widely prevalent vegetation type, throughout the year (Fig. 1b). Buffalo showed a broadly distributed use of habitat

types during the wet season, but concentrated strongly in the granitic region near the river during the dry season, most especially in the drier year (Fig. 1c). Both sable and zebra foraged mainly in upland regions of the landscape throughout the dry season (Fig. 2a). Buffalo concentrated in slope regions in the early dry season and made greater use of lowland near the river during the late dry season. Sable and zebra entered this lowland only to drink from pools in the river. http://www.selleckchem.com/products/Y-27632.html The foraging areas of this website zebra were usually more open with shorter trees than those occupied by sable (Fig. 2b). However, tree canopy cover and height in the foraging areas of buffalo were very similar to those for sable. Grass height

in foraging areas was generally in the range 41–80 cm for all three grazers, with no seasonal variation (Fig. 2c). The grassland tended to be greener than in the foraging areas of sable than in those of zebra and buffalo in the early dry season, but this distinction fell away during the late dry season when very little green grass remained (Fig. 2d). The model incorporating both grass greenness

and tree canopy cover best distinguished the foraging areas of sable from those zebra, although the model with greenness replaced by season was almost equally supported (Table 1a). For the sable–buffalo comparison, the best supported model included only grass greenness as a distinguishing feature, but with some support for an interaction with season (Table 1b). Either tree cover or topography was the most strongly supported distinction between the foraging areas of zebra and buffalo (Table 1c). Acceptance of the grass at feeding sites was more strongly influenced by grass greenness for sable than was the case for both zebra (G2 = 91.6, d.f. = 3, P < 0.001) and buffalo MCE公司 (G2 = 116.0, d.f. = 3, P < 0.001) (Fig. 3a). Zebra appeared somewhat indifferent to distinctions in greenness in their grass acceptance throughout the dry season, while buffalo showed an inconsistent response to grass greenness in the early dry season, and foraged solely in sites containing little or no green grass during the late dry season. During the late dry season, sites containing grass that was more than 10% green were present only in the feeding sites of sable. Sable differed from both zebra (G2 = 94.08, d.f. = 2, P < 0.001) and buffalo (G2 = 43.96, d.f. = 2, P < 0.001) in the influence of grass height on acceptance.

The diet of celiac, all patients responded clinically applied. Results: Colonoscopy results were all in patients with celiac, celiac disease patient colonoscopic Dorsomorphin order examination in patients with celiac biopsy results lympho plasmocytes infiltrate at the lamina propria was determined by pathological examination. Conclusion: As a result of the increasing frequency and is accepted as a major health problem all over the world primarily in celiac disease. Physicians and other health care providers, as well as the food industry

should maintain their work as more and more knowledgeable and well-equipped. This is as a result of mucosal sampling in the diagnosis of celiac disease and

gluten-free diet after the implementation of the follow-up or diagnostic purposes can be used as a different method. Key Word(s): 1. Celiac disease; 2. Colonic infiltration; 3. Lympho plasmocytes; Presenting Author: ZHENZHEN LIU Additional Authors: JIE LIANG, BIAOLUO WANG, JIAYUN LIU, KAICHUN WU Corresponding Author: ZHENZHEN LIU Affiliations: The Fourth Military Medical University Objective: To evaluate the clinical diagnostic value of T-cell spot of tuberculosis test (T-SPOT.TB) in tuberculous peritonitis EPZ6438 (TBP). Methods: TBP patients (n = 102) and non-TBP patients (n = 27) were detected the T-SPOT.TB in peripheral blood, 18 patients were detected the T-SPOT.TB in ascites and 82 patients were detected

the anti-TB antibody at the same time. Results: After the stimulus of ESAT-6 and CFP-10, the T cells gamma interferon spots forming cells (SFCs) in blood for TBP group were (153.40 ± 190.45)/10∧6peripheral MCE公司 blood mononuclear cell (PBMC) and (153.65 ± 217.23)/10∧6PBMC, respectively. After the stimulus of ESAT-6 and CFP-10, the T cells gamma interferon spots forming cells (SFCs) in blood for non-TBP group were (24 ± 45.55)/10∧6PBMC and (18 ± 40.19)/10∧6PBMC, respectively. The SFCs of T-SPOT.TB showed significant difference between the two groups (P < 0.05). The sensitivity and specificity of T-SPOT.TB in blood were 94.1%(96/102) and 70.4%(20 /27). The sensitivity and specificity of anti-TB anti-body in blood were 28%(23/82) and 75%(6/8). The sensitivity between T-SPOT.TB and anti-TB antibody showed significant difference (P < 0.05). The sensitivity and specificity of T-SPOT.TB in ascites were 90%(9/10) and 50%(4/8). The SFCs of T-SPOT.TB in ascites were higher than blood in 90%(9/10) of patients in two group. Conclusion: The application of T-SPOT.

3,4 In 2009, GMA was also accepted as

an adjunct therapeutic strategy for active CD patients according to the superior results obtained from a nationwide multicenter trial.5 It is now therefore an appropriate opportunity to upgrade and summarize our current understandings and/or future perspectives of this unique non-pharmacological and non-surgical strategy of CAP for IBD patients. Filtration leukocytapheresis and GMA are the most used CAP techniques for intractable UC patients with acute flare. According to a national survey in Japan, the total number of UC patients has been expanding gradually, and it has now reached to over 100 000. Among them, almost 50% of patients have been facing active flare more severely than moderate; and, approximately 30% of them were diagnosed as “intractable”, meaning either treatment resistance or dependent characteristics C646 research buy for conventional steroid therapy (Fig. 1). Patients with intractable active UC flare are potential candidates for applying an adjunct strategy, including immunosuppressant, biologics, and CAP. We have developed both LCAP and GMA, and the current tasks

for them should be to determine the appropriate therapeutic regimen in order to obtain the maximum clinical efficacy of these unique non-pharmacological and non-surgical interventions. Filtration leukocytapheresis.  Filtration leukocytapheresis is performed using a specially designed leukocyte removal MLN0128 research buy column, Cellsorba EX (Asahi Kasei Kuraray Medical, Tokyo, Japan), set on a simple one-way hemofiltration circuit.3,6,7 A roller pump drains the patient’s peripheral blood from an antecubital vein under constant flow rate of 50 mL/min. An optimal amount of Nafamostat mesilate (NM; Futhan; Torii Pharmacology, Tokyo, Japan) or heparin is mixed with saline and added to the drained peripheral whole blood as anticoagulant before infusion into the column (Fig. 2). Polyester non-woven leukocyte removal filter was installed into the polycarbonate outer shell

of Cellsorba. Approximately 35% of platelets are expected to be removed by LCAP from processed peripheral blood, together with almost 100% of granulocytes and monocytes and 64% of lymphocytes (Fig. 3a).7 MCE公司 Adsorptive granulocyte/monocyte apheresis.  Granulocyte/monocyte apheresis is performed with the Adacolumn (JIMRO, Takasaki, Japan). The circuit diagram for GMA is almost the same as that of LCAP. Peripheral whole blood drained from the patient’s body is passed at 30 mL/min, a flow speed created by an external roller pump with optimal amount of NM or heparin as an anti-coagulant. The Adacolumn is filled with cellulose acetate beads, which serve as the column adsorptive leukocytapheresis carriers. The carriers in the column selectively adsorb about 65% of granulocytes, 55% monocytes/macrophages and a smaller fraction of lymphocytes.

3,4 In 2009, GMA was also accepted as

an adjunct therapeutic strategy for active CD patients according to the superior results obtained from a nationwide multicenter trial.5 It is now therefore an appropriate opportunity to upgrade and summarize our current understandings and/or future perspectives of this unique non-pharmacological and non-surgical strategy of CAP for IBD patients. Filtration leukocytapheresis and GMA are the most used CAP techniques for intractable UC patients with acute flare. According to a national survey in Japan, the total number of UC patients has been expanding gradually, and it has now reached to over 100 000. Among them, almost 50% of patients have been facing active flare more severely than moderate; and, approximately 30% of them were diagnosed as “intractable”, meaning either treatment resistance or dependent characteristics R428 in vitro for conventional steroid therapy (Fig. 1). Patients with intractable active UC flare are potential candidates for applying an adjunct strategy, including immunosuppressant, biologics, and CAP. We have developed both LCAP and GMA, and the current tasks

for them should be to determine the appropriate therapeutic regimen in order to obtain the maximum clinical efficacy of these unique non-pharmacological and non-surgical interventions. Filtration leukocytapheresis.  Filtration leukocytapheresis is performed using a specially designed leukocyte removal SP600125 column, Cellsorba EX (Asahi Kasei Kuraray Medical, Tokyo, Japan), set on a simple one-way hemofiltration circuit.3,6,7 A roller pump drains the patient’s peripheral blood from an antecubital vein under constant flow rate of 50 mL/min. An optimal amount of Nafamostat mesilate (NM; Futhan; Torii Pharmacology, Tokyo, Japan) or heparin is mixed with saline and added to the drained peripheral whole blood as anticoagulant before infusion into the column (Fig. 2). Polyester non-woven leukocyte removal filter was installed into the polycarbonate outer shell

of Cellsorba. Approximately 35% of platelets are expected to be removed by LCAP from processed peripheral blood, together with almost 100% of granulocytes and monocytes and 64% of lymphocytes (Fig. 3a).7 medchemexpress Adsorptive granulocyte/monocyte apheresis.  Granulocyte/monocyte apheresis is performed with the Adacolumn (JIMRO, Takasaki, Japan). The circuit diagram for GMA is almost the same as that of LCAP. Peripheral whole blood drained from the patient’s body is passed at 30 mL/min, a flow speed created by an external roller pump with optimal amount of NM or heparin as an anti-coagulant. The Adacolumn is filled with cellulose acetate beads, which serve as the column adsorptive leukocytapheresis carriers. The carriers in the column selectively adsorb about 65% of granulocytes, 55% monocytes/macrophages and a smaller fraction of lymphocytes.

Coronary artery calcium (CAC) scoring has been extensively studied as a powerful, non-invasive tool for cardiovascular risk assessment in the general population. The aim of this study is to investigate whether CAC scoring could predict obstructive CAD in asymptomatic LT candidates with liver cirrhosis (LC). Methods: This study included 850 consecutive cirrhotic patients who underwent computerized coronary angi-ography with CAC measurement using

the Agaston method as a pre-LT workup. None of these patients had a previous CAD history. Obstructive CAD was JNK inhibitor defined as ≥50% of lumi-nal narrowing in any artery on computerized angiography. The association between CAC score and obstructive CAD was analyzed using the Pearson correlation method, logistic regression and area under the receiver operating characteristic CX-5461 supplier curve (AUROC) analyses. Results: The mean CAC score of all patients was 90.0 (range, 0-4411.4). The CAC score was 0 for 535 patients (62.9%), 1-100 for 191 (22.5%), 101-400 for 74 (8.7%), and >400 for 50 (5.9%). Obstructive

CAD was identified in 72 patients (8.5%). The mean CAC score significantly differed between patients with and without obstructive CAD (633.6 vs. 39.6; P<0.05). The prevalence of obstructive CAD increased with the CAC score (1.7% for 0, 5.8% for 1-100, 25.7% for 101-400, and 66.0% for >400). The CAC score was significantly correlated with the grade of coronary stenosis (r=0.71; P<0.05). The CAC score showed excellent performance for predicting obstructive CAD with an AUROC value of 0.88. The best cut-off CAC score was 38.8 for

obstructive CAD with a sensitivity of 83% and a specificity of 86%. In multivari-ate 上海皓元 analysis, a CAC score at a cut-off of 38.8 was an independent predictor for obstructive CAD (adjusted odds ratio[OR], 23.9; P<0.05). Older age, male sex, a current smoker, hypertension, diabetes, and alcoholic LC were significantly associated with a CAC score above 38.8 (adjusted OR, 1.07, 3.27, 1.59, 1.54, 1.79, and 2.17; Ps<0.05), as were neither liver function and coagulation parameters nor viral hepatitis affect the score. Conclusion: Our data indicate that the CAC score is an accurate tool for predicting subclinical obstructive CAD in cirrhotic subjects. Traditional cardiovascular risk factors, together with alcoholic LC, were closely associated with higher CAC score.

PCR primers tailed with common sequences (e.g. m13) facilitate DNA sequence reaction set up, enabling any amplicon from any gene to be sequenced using single forward and reverse primers. Mutation scanning techniques such as high

resolution melt curve analysis (HRM) [23] are also amenable to high throughput analysis. Robotic sample processing eliminates inter-user variation in pipetting, resulting in more consistent DNA sequence whilst enhancing sample throughput. Following DNA sequencing, application of sequence analysis software enables rapid identification of variants that differ from the reference sequence (RefSeq). Large deletions and duplications can be identified using multiplex ligation-dependent probe amplification (MLPA) [24,25], micro-array or see more mutation-specific gap-PCR [26]. F8 intra-chromosomal inversions CP-868596 supplier amplified through long or inverse PCR can be analysed using gel electrophoresis

or, for smaller amplicons, by fragment sizing using a DNA sequencer. Previously ‘missing’ deep intronic mutations that affect splicing can be detected by reverse transcription (RT)-PCR from mRNA, gel electrophoresis and DNA sequencing [27,28]. Sequence-variant interpretation software integrates pathogenicity prediction tools for amino acid substitutions and for splice variants along with literature searching for previously reported variants. For amino acid substitutions, these are based on different algorithms for their potential impact on protein structure/function using assessment of physical properties, along 上海皓元医药股份有限公司 with their evolutionary conservation. Mutations potentially affecting splicing are analysed by panels of algorithms analysing presence and relative strength of splice motifs. Locus-specific mutation databases, the Human Gene Mutation Database (HGMD) [29], and the single nucleotide polymorphism database (dbSNP) [30] catalogue and help facilitate pathogenicity interpretation of previously reported variants.

Newly recognized sequence variants underneath PCR amplification primers can be sought with each new release of dbSNP using tools including SNPCheck [31]. Primers can then be redesigned where necessary using online software to prevent mono-allelic amplification resulting in allele dropout and mutations potentially being missed. [32] Reference genetic materials are available for common mutations through the UK National Institute for Biological Standards and Control (NIBSC) [33] and can be used to validate assay performance. LIMS use can facilitate reporting of the results of genetic testing to the referring clinician through ready availability of all patient documentation on the computer screen and through integration of standard report templates with specific details of testing undertaken and its results.

To test this hypothesis, we performed ubiquitination assays. Ubiquitination PI3K Inhibitor Library screening levels of AIB1 protein were dramatically decreased in the presence of HBx in both 293T and HepG2

cells, demonstrating that HBx inhibits the ubiquitination of AIB1 (Fig. 3A,B). To determine whether HBx could interact with AIB1 to inhibit the ubiquitination of AIB1, Flag-tagged AIB1 and HA-tagged HBx were coexpressed in HepG2 cells, and Co-IP assays were performed. Anti-Flag antibodies, but not control IgG, immunoprecipitated HBx from cell lysates (Fig. 3C, left panel). Reciprocally, anti-HA antibodies could also immunoprecipitate AIB1 from cell lysates (Fig. 3C, right panel). To further evaluate whether the interaction between AIB1 and HBx could occur in human HCC specimens, we performed Co-IP assays in three human HCC specimens (21T, 30T, and 32T), which showed a high expression of both AIB1 and HBx. Anti-AIB1 antibodies, but not control IgG, could coimmunoprecipitate HBx protein in these samples (Fig. 3D). Reciprocally, anti-HBx antibodies could coimmunoprecipitate AIB1 protein. These data Selleckchem Cobimetinib suggest that the interaction between HBx and AIB1 might be involved in the HBx-mediated inhibition of AIB1 ubiquitination. It has been reported that SCFFbw7 E3 Ub ligases can specifically and effectively mediate the ubiquitination and degradation of its substrates.18 AIB1 has

been identified as one of the substrates of Fbw7α.19 Therefore, we tested whether HBx could inhibit the Fbw7α-mediated ubiquitination and degradation of AIB1. Western blotting analysis revealed that Fbw7α down-regulated AIB1 in a MCE dose-dependent manner; however, the Fbw7α-mediated down-regulation of AIB1 was dramatically inhibited by HBx (Fig. 4A). To further determine whether HBx could inhibit the Fbw7α-mediated ubiquitination of AIB1, we performed ubiquitination assays. The Fbw7α-mediated increase of AIB1 ubiquitination was significantly inhibited by HBx (Fig. 4B). Because HBx can interact with AIB1 protein, it is possible that HBx inhibits the Fbw7α-mediated ubiquitination and degradation of AIB1 through disruption

of the interaction between AIB1 and SCFFbw7α. To test this hypothesis, we performed Co-IP assays to examine the interaction between Fbw7α, AIB1, and HBx. Western blotting analysis revealed that Fbw7α interacted with AIB1 in the absence of HBx (Fig. 4C, lane 5); however, the interaction between Fbw7α and AIB1 was dramatically reduced in the presence of HBx (Fig. 4C, lane 6 versus lane 5), demonstrating that HBx inhibits the interaction between Fbw7α and AIB1. It is reported that phosphorylations of S505 and S509 in AIB1 are important for Fbw7α to regulate AIB1 turnover, and the down-regulation effect of Fbw7α on AIB1 is attenuated or completely abolished when either S505 or S509 is mutated to alanine (A).19 We found that HBx significantly up-regulated the protein level of wild-type (WT) AIB1, as expected (Fig.

The good-response genotype (C/C Enzalutamide in vivo rs12979860 or T/T rs8099917) was strongly

associated with an increased rate of sustained virological response despite the addition of a directly acting antiviral agent. This suggests that patient IL-28B genotype will remain relevant in the dawning era of specifically targeted antiviral therapy for hepatitis C virus because a combination with peg-IFN and RBV is required to restrict the development of antiviral resistance. It will, therefore, be important to consider IL-28B genotype in clinical development programs; because of the population frequency of the good-response IL-28B variant and its association with rapid viral decline during peg-IFN therapy,2 it is possible for small early-phase efficacy trials CH5424802 datasheet to be confounded by an imbalance in the IL-28B genotype across treatment arms. We statistically modeled the probability of an imbalance in the good-response IL-28B variant (C/C rs12979860) between treatment arms for three hypothetical situations: a phase

1 trial (n = 60), a phase 2a trial (n = 120), and a phase 2b trial (n = 240). Each involved three randomized arms (Fig. 1). The probability of an imbalance in one treatment arm of ±10% (<23% or >43% when the C/C genotype frequency was assumed to be 33%2) was 31%, 18%, and 6% for the phase 1, 2a, and 2b trials, respectively, and the probability of an imbalance in one treatment arm of ±20% was 10%, 0.4%, and <0.01% for the phase 1, 2a, and 2b trials, respectively. We assumed a Caucasian population for this analysis; the inclusion of other ethnic groups would be expected to increase the risk of sampling error.2 We then modeled

the implications of such an imbalance for the primary outcome of viral load 上海皓元医药股份有限公司 reduction at week 4 in studies combining a direct antiviral agent with peg-IFN and RBV (Table 1). An imbalance in the IL-28B genotype of 10% to 20% could lead to differences in HCVRNA reduction of 0.2- to 0.5-log10 IU/mL between treatment arms due to peg-IFN alone. This has great relevance for dose-finding studies in which the dose-related antiviral potency must be weighed against toxicity. In the setting of more extreme mismatching (e.g., in a mixed-ethnicity cohort), confounding by IL-28B genotype might even affect the decision to advance a compound from proof of concept to the next stage of clinical development. Indeed, Anadys Pharmaceuticals recently reported an imbalance in the frequency of the C/C genotype that confounded the week 12 results of a phase 2 trial (the C/C genotype frequency was 21% in the active treatment arms and 56% in the control arm).

The good-response genotype (C/C Dinaciclib rs12979860 or T/T rs8099917) was strongly

associated with an increased rate of sustained virological response despite the addition of a directly acting antiviral agent. This suggests that patient IL-28B genotype will remain relevant in the dawning era of specifically targeted antiviral therapy for hepatitis C virus because a combination with peg-IFN and RBV is required to restrict the development of antiviral resistance. It will, therefore, be important to consider IL-28B genotype in clinical development programs; because of the population frequency of the good-response IL-28B variant and its association with rapid viral decline during peg-IFN therapy,2 it is possible for small early-phase efficacy trials LY294002 purchase to be confounded by an imbalance in the IL-28B genotype across treatment arms. We statistically modeled the probability of an imbalance in the good-response IL-28B variant (C/C rs12979860) between treatment arms for three hypothetical situations: a phase

1 trial (n = 60), a phase 2a trial (n = 120), and a phase 2b trial (n = 240). Each involved three randomized arms (Fig. 1). The probability of an imbalance in one treatment arm of ±10% (<23% or >43% when the C/C genotype frequency was assumed to be 33%2) was 31%, 18%, and 6% for the phase 1, 2a, and 2b trials, respectively, and the probability of an imbalance in one treatment arm of ±20% was 10%, 0.4%, and <0.01% for the phase 1, 2a, and 2b trials, respectively. We assumed a Caucasian population for this analysis; the inclusion of other ethnic groups would be expected to increase the risk of sampling error.2 We then modeled

the implications of such an imbalance for the primary outcome of viral load MCE公司 reduction at week 4 in studies combining a direct antiviral agent with peg-IFN and RBV (Table 1). An imbalance in the IL-28B genotype of 10% to 20% could lead to differences in HCVRNA reduction of 0.2- to 0.5-log10 IU/mL between treatment arms due to peg-IFN alone. This has great relevance for dose-finding studies in which the dose-related antiviral potency must be weighed against toxicity. In the setting of more extreme mismatching (e.g., in a mixed-ethnicity cohort), confounding by IL-28B genotype might even affect the decision to advance a compound from proof of concept to the next stage of clinical development. Indeed, Anadys Pharmaceuticals recently reported an imbalance in the frequency of the C/C genotype that confounded the week 12 results of a phase 2 trial (the C/C genotype frequency was 21% in the active treatment arms and 56% in the control arm).