This drug has a shorter half-life (2-5 h) than bevacizumab and its daily administration could be better controlled to limit toxicity [22]. Axitinib used in a phase II trial for advanced NSCLC demonstrated an increased one-year survival rate with manageable toxicities [17] and [22] and was well tolerated when combined with platinum doublets chemotherapy [23]. The role of angiogenesis in the progression and prognosis of NSCLC

and its targeting by various new anti-angiogenic drugs either alone or combined with conventional chemotherapy for NSCLC are under extensive clinical investigation [24], [25], [26] and [27]. However, the combination of anti-angiogenic drugs with RT, which is the conventional treatment for stage III inoperable find more NSCLC, has not been explored. The goal of the current study was to explore whether axitinib could improve the efficacy of RT for NSCLC using a pre-clinical model of orthotopic lung carcinoma. We hypothesized that an anti-angiogenic drug, Selleck GDC-941 given at doses which trim inefficient tumor vessels and regularize blood flow, could improve oxygenation in the tumor microenvironment

and enhance RT efficacy for locally advanced NSCLC. Alternatively, higher doses of anti-angiogenic drugs resulting in a cytostatic effect could enhance the cytoreductive effect of RT. Using these concepts, we have previously demonstrated that a dose of sunitinib, which regularized tumor vessels and blood flow, enhanced the efficacy of chemo- and radio-therapies for metastatic RCC in an orthotopic RCC pre-clinical mode [28], [29] and [30]. However, the dose of sunitinib used in these studies was reduced to avoid toxicity to the vasculature HSP90 of normal tissues [28], [29] and [30]. We now report studies confirming that axitinib is a potent and safe anti-angiogenic drug that significantly enhances the efficacy of lung irradiation in an orthotopic xenograft model of lung carcinoma. This combined therapy is well tolerated with no further increase

in radiation-induced injury or vascular damage in lung tissue but quite the opposite effect was observed suggesting a radioprotective effect. The human non-small cell lung carcinoma (NSCLC) A549 (purchased from ATCC) was cultured in F-12 K culture medium containing 7% heat-inactivated fetal bovine serum with supplements. A549 cells, at 2×10 [6] in 200 μl HBSS, were injected i.v. in the tail vein of 5-6 week old female Hsd Athymic Nude-Foxn1nunu/nu nude mice (Harlan, Indianapolis, IN) [31]. Mice were housed and handled under sterile conditions in facilities accredited by the American Association for the Accreditation of Laboratory Animal Care (AAALAC). The animal protocol was approved by Wayne State University Animal Investigation Committee (IACUC). Three anesthetized mice, in jigs, were positioned under a 6.

The cleavage of the azo bond by the oxidative process was confirmed by the results obtained with the electrochemical oxidation experiments. It can be seen in Fig. 3 that the band characteristic of the chromophore group of DR1 (at 510 nm) decreased during the electrolysis when performed at +1.5 V for up to 50 min. Concomitantly, a new peak was observed click here at 640 nm, due to the formation of stable radicals and change in color. After 90 min of electrolysis, the total removal of the bands due to the chromophore group, total discoloration and loss of the extra bands at 640 nm were verified (Fig. 3). This indicates that

the spectroelectrochemical technique detected the radical as an intermediate product, which vanished in the presence of oxygen or after a long electrolysis time. According to this finding, sulfate 2-[(4-aminophenyl)ethylamino]-ethanol monohydrate with a retention time (tR) of 10.0 min and

nitrobenzene (tR = 12.0 min), in a proportion of 6% and 7% respectively, were detected after 2.5 h of oxidation by controlled potential electrolysis ( Fig. 4). With the objective of determining whether this effect also occurred under reducing conditions, the experiments were repeated monitoring the reduction of 3.18 × 10−4 mol L−1 in 0.01 mol L−1 DMSO/TBABF4 slightly acidified with acetic acid, using a potential of −1.5 V. The UV–Vis spectra recorded simultaneously during the reduction of Red 1 indicated a decrease in the band at 510 nm up to 60 min, but there was no extra peak at 640 nm (Fig. 5). The DR1 dye solution (3.18 × 10−4 mol L−1 in 0.01 mol L−1 DMSO/TBABF4) Selleck FK228 was also subjected to 2.5 h reduction using

controlled potential electrolysis, the solution being previously deaerated by bubbling in N2 (99.7% purity) for 10 min. The reaction was monitored every 30 min and the band corresponding to the chromophore group was totally suppressed after only 2 h of electrolysis. However, even under these conditions there was no evidence of the formation of intermediate stable radicals during the reduction process of the nitro group of the DR1 dye. Thus the electrolyzed product was submitted to extraction and identified by HPLC/DAD, which indicated the formation of the same aromatic amine (sulfate 2-[(4-aminophenyl)ethylamino]-ethanol monohydrate) previoulsy Leukotriene-A4 hydrolase detected in a proportion of 9%. Nitrobenzene was not detected under these conditions. Using GC/MS 4-nitro-benzamine was also detected, after both the oxidation and reduction processes, confirming the generation of aromatic amines after cleavage of the bond. According to the mass spectra corresponding to the peaks, the peaks tR = 13.576 min and 13.513 min ( Fig. 6A and B, respectively) are related to the substance 4-nitro-benzamine ( Fig. 7). In addition, after an analysis of the reduction products, 2-(ethylphenylamino)-ethanol was also detected. Table 1 summarizes the products detected after the oxidation and reductions reactions.

Very little demographic information was provided about the people (physicians, nurses, pharmacists, and so forth) who received the interventions and in most studies it is not clear how many prescribers were involved. The studies ranged in size from 21 to 7000; approximately 19,300 people with dementia were included in total (information not provided in all studies). Descriptions of the interventions used in the studies are shown in Table 3. We grouped studies according to intervention type using

4 categories: educational programs (n = 11 studies), in-reach services (n = 2 studies), medication review (n = 4 studies), and multicomponent interventions (n = 5 studies). The EPOC Data Collection Checklist includes PLX4032 mouse a taxonomy of intervention components grouped under 4 headings: professional, organizational, structural, and regulatory.16 The interventions within studies of educational programs14, 18, 19, 20,

23, 24, 25, 29, 30, 31 and 32 consisted mainly of professional components, such as educational meetings, distribution of educational materials, and educational outreach. In-reach services21 and 26 contained mainly organizational and structural components. Studies containing the most variety were those in the medication review22, 33, 34 and 35 Sunitinib concentration and multicomponent intervention groups27, 28, 36, 37, 38 and 39 incorporating educational, organizational, structural, and

regulatory interventions. In many cases, there was insufficient information provided in the article to replicate the intervention in another setting. Using the EPOC Data Collection Checklist classification, the number of intervention components per study ranged from 1 to 7; most studies consisted of 3. The most frequently Forskolin concentration used intervention component was educational outreach (14 studies), and this was evident across all 4 types of intervention. Educational outreach was defined as the use of a trained person who met with providers in their practice settings to give information with the intent of changing the provider’s practice. Assessment of the quality of each included study is shown in Table 4. The global assessment of just over a third of the studies was moderate or strong. The main areas of weakness were in the collection of primary outcome data and in the reporting of withdrawals and dropouts. In most of the studies, the outcome assessor was aware of the intervention status of participants and the study participants (prescribers) were aware of the research question. Although data on prescribing rates were taken from patient and pharmacy records in many cases, the data-collection process was performed by one individual with no procedure for checking accuracy. Furthermore, the data-collection tool was often not described, precluding judgment on the validity of the measure.

Each panelist received 6 h of training sessions and practice in soymilk evaluation. During the training, panelists evaluated and

discussed soymilk sensory attributes by comparing to cv. ZH13. Specific attributes, attribute definitions, and references were developed by the panelists (data not shown). Panelists compared six parameters—including colour and appearance, aroma, sweetness, thickness in the mouth, smoothness in the mouth, and overall acceptability—and assigned a score to each sample based on a 7-point hedonic scale (1–7) for soymilk flavour sensory evaluation: 1 = ‘strongly disliked’; 2 = ‘moderately disliked’; 3 = ‘slightly disliked’; 4 = ‘indifferent’; 5 = ‘slightly liked’; 6 = ‘moderately liked’; and 7 = ‘strongly liked’ ( Robinson, Chambers, & Milliken, 2005). To adapt to a traditional taste style, the soymilk was kept at approximately learn more 70 °C before sensory evaluation. The analysis of variance (ANOVA) indicated

that the panel and panelists could consistently use the attributes to differentiate the soymilk samples. For the soymilk flavour evaluation, the basic panel procedures followed the previous method (Chambers, Jenkins, & McGuire, 2006). The panel tasted one sample at a time. The flavour and mouth feel attributes were recorded 60 s after swallowing. The panel openly discussed each soymilk sample to reach Selleck Verteporfin a consensus concerning the flavour and mouth feel

properties. The protein and oil content could be estimated by near-infrared spectroscopy (Hymowitz, Dudley, Collins, & Brown, 1974). In this study, 50 g of soybean seeds for each sample were analysed by transform near-infrared absorption spectroscopy (Bruker Fourier, Germany). The spectrum value of each sample represented the average value of triplicate and the absorption ranged from 4000 to 8000 cm−1. The collected spectra were transferred to the protein and oil content by the Quant 2 method of Bruker’s OPUS 4.2 software. It is reported 11S/7S ratio can be used as a criterion of indirect selection for high quality protein (Sharma, Kaur, Goyal, & Gill, 2014). For determination of the 11S/7S ratio, the storage protein subunits glycinin (11S) and β-conglycinin triclocarban (7S) were quantified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) (Bradford, 1976). Ten milligrams of soybean flour for each sample were extracted with 500 μL extraction solution (0.05 M Tris buffer, pH 8.0, 0.01 M β-mercaptoethanol, and 2% SDS) for 1 h at 4 °C. Samples were then centrifuged at room temperature at 12,000 rpm for 15 min. The supernatant contained the total soybean proteins. Next, 5.0 μL of supernatant was loaded onto a gradient gel containing 5–12% polyacrylamide.

There was no improvement in pVO2 at 6 months. There was a low rate of heart failure hospitalization in these patients. Over 12 months, 3 of the 20 implanted patients (15%) had 5 Clinical buy Everolimus Events Committee–adjudicated

heart failure hospitalizations. Few of these occurred during active therapy, and C-Pulse System nonadherence appeared to be related to most of these heart failure hospitalizations; specifically, 2 of these 3 patients were nonadherent (utilizing the system <30% of the time) in the weeks before their heart failure hospitalizations. Over 12 months, there were 40 non–heart failure related hospitalizations in 19 patients. Of these, 10 were related to PIL issues in 9 patients (45%), 11 to exit site infections in 8 patients (40%), 7 to other infections (urinary tract infection, sternal wound, pneumomediastinum, peripherally inserted central catheter) in 4 patients (20%), and 12 to other conditions (e.g., atrial fibrillation, respiratory failure, disseminated intravascular coagulation) in 9 patients. The results of this feasibility GSI-IX solubility dmso study suggest that the C-Pulse

System may be safe and effective in patients with moderate to severe heart failure. A majority of patients showed improvements in NYHA functional class and QoL scores, and statistically significant improvements in mean change from baseline to 6 and/or 12 months were demonstrated for NYHA functional class, QoL scores, and the 6MWD. Considering that this feasibility study was neither designed nor powered to demonstrate statistically significant improvements in any of the efficacy

measurements, these findings should merely be considered as preliminary indicators of the potential efficacy of the C-Pulse System. However, in this context, the magnitude of these improvements is clinically meaningful when compared to prior drug and device trials in heart failure 3 and 4, and occurred on top of ongoing treatment with optimal heart failure drug and electrophysiological device therapies. Further support for these preliminary efficacy signals includes the successful weaning of inotropes in all patients receiving inotropes at baseline and the reduction in diuretic requirements in 6 patients (30%), implying improved cardiac output and peripheral perfusion. There was no increase in diuretics for any of the patients in the study. These findings require confirmation in an C-X-C chemokine receptor type 7 (CXCR-7) adequately powered randomized controlled trial. From the safety standpoint, the composite adverse event assessment was dominated by the incidence of manageable exit site infections, which might be mitigated in the future by recently developed strategies for better drive line fixation and management. There were no neurological events, myocardial infarctions or periprocedural mortality. For the 12-month period, there was 1 device-related death reported, attributed to complications arising from a sternal wound infection in a patient who underwent repeated sternotomies and attempted sternectomy.

To add weight to his arguments, Gazzaniga claims (in a review) that scientific advances in the study of brain mechanisms do not undermine the foundations of the action decision mechanism underlying moral responsibility; so it is time to get over the idea of FW and move on (Gazzaniga, 2012). From a different perspective, Dennet claims that the conclusion that FW does not exist, might means “bad news” (Dennett, 2011). The public generally considers philosophy to be fairly ineffectual in everyday life, however, FW issue matters to people, especially if we consider its role in determining moral behaviour, then philosophers should intervene clearly and

unambiguously on the FW issue. Since people may think that FW is a myth, the idea that “my mind made do it” could be a convenient way of passing the buck and escaping blame and penalty. The law presumes ‘moral competence’ of an individual in order to judge Tariquidar datasheet him, then, the main question is whether a robotic mind may acquire a sense of agency and responsibility in order to understand and accept reward or blame. The wiring of our brain circuits provides us

with the cognitive selleck compound ability to bring about the necessary moral competences. Thus, the moral imperative of scientific progress is to discriminate clearly between the circumstances in which an individual can and cannot be considered properly responsible for his action. Today, neuroethical studies tend to disregard the FW issue, so that whether science demonstrates FW is an illusion or not is irrelevant. This consideration, however,

opens up another aspect of mind/body duality. According to TBM, the conscious agent thinks he possesses FW, and thIs belief, though illusionary, is a real and unavoidable part of the individual, thus, the importance of TBM lies in the fact that the first- and third-person perspectives of the role of the conscious agent in intentional action have the same dignity; they serve as Galeterone tools to understand the mechanism of human cognition. In this mechanism, we do not lose sight of the fundamental role of FW illusion. In this perspective, the fundamental question is: “Is the CM a sheaf of experiences collected and organised by some type of automatism in the brain, or is it the manifestation of a spirit?” If duality does exist it is easier to discuss moral responsibility; however, there is an inherent contradiction in the belief in the automaticity of the brain in intentional actions (FW illusion) and the self-attribution of free responsibility in ethical decisions. Alternatively, we wonder if we can trust the intentions that determine personal and social behaviour if we believe in TBM (see point 3). Conscious FW is invoked to attribute to an individual the responsibility of an intentional action. A man can be liable by law only if his actions have been performed with conscious intentions (mens rea) ( Morawetz, 1980).

ex. Olaparib mouse Loud.) and at low elevations in the southern portion of the region, ponderosa pine (Pinus ponderosa Dougl. ex P. & C. Laws). In a study of Douglas-fir growth in BC, Chen et al. (2010) found that radial growth trends across all interior regions was positively correlated with precipitation in the fall of the previous year and in the current growing season, while radial growth was negatively correlated with temperature of the current growing season, suggesting that water stress is an important parameter limiting radial growth. Griesbauer and Green (2010) found that Douglas-fir radial

growth was strongly correlated with previous July to current June precipitation, with moisture sensitivity most pronounced at the dry southern margins of the region. The radial growth of ponderosa pine is correlated positively to previous August and current July precipitation ( Watson and Luckman, 2002), and negatively to current June temperature ( Campbell et al., 2006), while radial growth of lodgepole pine is correlated positively to previous MEK activation July and current June–July precipitation ( Watson and Luckman, 2002, Lo et al., 2010 and McLane et al., 2011). The negative radial growth correlations exhibited by all three tree species to summer temperature in interior BC suggests that

increased evaporative losses and water stress during high temperature intervals are detrimental to tree growth (Watson and Luckman, 2002). In mid- to low elevation interior ecosystems, tree-ring variability is primarily related to factors affecting water supply, especially precipitation, indicating that tree growth is limited by moisture availability in the previous and current growing seasons (Watson and Luckman, 2002, Campbell

et al., 2006, Littell et al., 2008, Chen et al., 2010, Griesbauer and Green, 2010, Lo et al., 2010 and McLane et al., 2011). One difficulty in reconstructing IMP dehydrogenase WSB outbreaks in the Cariboo Forest Region is the limited availability of long-lived non-host Pinus trees. The recent mountain pine beetle outbreak affected 18.1 million hectares of mature forest in BC ( BCMFLNRO, 2012), decimating Pinus species across their geographic distribution. As a consequence, it was necessary to access previously collected tree-ring data to construct non-host chronologies for our study. Lodgepole pine chronologies were archived at the Pacific Forestry Centre ( Alfaro et al., 2004) and at the University of British Columbia Tree-Ring Laboratory ( Daniels and Watson, 2003). Ponderosa pine chronologies were archived at the International Tree-Ring Data Bank (ITRDB), the University of British Columbia Tree-Ring Laboratory ( Daniels and Watson, 2003), and at the University of Victoria Tree-Ring Laboratory ( Campbell et al., 2005 and Campbell et al., 2006). While the convention in tree-ring based reconstructions of WSB is to collect host and non-host chronologies from the same or adjacent forest stands (e.g., Swetnam and Lynch, 1989), as has been the case in other studies ( Boulanger et al.

If a client calls that has violated the 24-hour rule, clinicians must realize that if medical assistance is needed then this is the priority. As Mintz (1961) stated, the risk of reinforcing future unskillful behaviors should not be prioritized over the risk of a client SB431542 in vivo dying. However, as much as possible the therapist should triage the medical attention to others who are less likely to be reinforcing. Thus, calling the EMS to assess and transport the client is less likely to reinforce the behavior than the therapist assessing the situation. Similarly, it is important that the therapist minimize any soothing responses, remain matter of fact, ensure that appropriate medical

attention is solicited and procured, and then end the call. Future therapy sessions may involve a behavior chain analysis that reveals to the client how they can become mindful of dysregulation before emotions reach a crisis level. The second target in

DBT telephone coaching is to assist clients in generalizing the skills that they are learning in treatment to everyday life (Linehan, 1993). During intense crises, clients with BPD often have difficulty accessing and applying information taught in the therapy context to the real world. Furthermore, clients Alpelisib with BPD can lack important interpersonal skills, often making their social environments challenging arenas to effectively navigate. The change-based skills in DBT, such as the interpersonal effectiveness skills and the emotion regulation skills, can be very useful for clients who are struggling with managing their emotions and/or interpersonal situations (Ben-Porath & Koons, 2005). Clients should be provided with examples of appropriate reasons to call for coaching (see Video). Some clients may be struggling with how to refuse a request from a friend while others may be struggling with feeling abandoned or hurt by a disagreement with a family member. Under these circumstances

telephone coaching provides an opportunity for clients to gain additional skills that they can then practice, in the moment, rather than after the fact. When clients traditionally have engaged in unskillful behaviors between sessions, these behaviors 4��8C are examined the following week in therapy, along with alternative skillful behaviors that could result in positive outcomes. By employing intersession phone coaching, therapists are able to maximize the principles of behaviorism. By having skills coaching immediately available, positive outcomes are maximized and the connection between skillful behaviors and positive outcomes are temporally linked and more immediately reinforced (Skinner, 1953). The following vignette provides an example of a phone coaching call in which the client is struggling with skills generalization. CLIENT: Hi. Okay, I am struggling right now with an issue with my brother and sister-in-law. I just don’t know what to do.

” (Garrett, 2007). There is a saying in Krio, the lingua franca of Sierra Leone, “mae we hush,” which is a term of condolence. The speaker offers condolences to the listener, while at the same time consoling him or herself for a shared loss. So for Khan, Fonnie,

their fellow healthcare workers fallen in the line of duty, and all those suffering from EVD in West Africa: mae we hush. Sheik Selisistat Humarr Khan is survived by his parents, son and daughter and 9 brothers and sisters. Mbalu Fonnie is survived by her mother, three sons and one daughter and four grandchildren. Readers who would like to make donations to a foundation established by the Khan family to help educate children orphaned by EVD may contact the corresponding author for information. The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, nor the U.S. Government. Dr. Bausch is a contractor employee of the U.S. Government. This work was prepared as part of his official duties. Title 17 U.S.C. §105 provides that ‘Copyright protection under this title is not available for any work of the United States Government’. Title 17 U.S.C. §101 defines a U.S. Government work as a work prepared by a military service

member or employee of the U.S. Government as part of that person’s official duties. The authors thank Mafudia Suaray for creative inputs and Cecilia Gonzales for administrative selleck products support. “
“Hepatitis C virus (HCV) is a single-stranded RNA virus and represents a major causative agent of chronic liver disease. Worldwide, 170 million people have a chronic HCV infection and are at risk to develop cirrhosis, leading to clinical complications such as hepatocellular carcinoma (HCC) (Hajarizadeh et al., 2013 and Lauer and Tacrolimus (FK506) Walker, 2001). The

aim of chronic hepatitis C treatment is to achieve a sustained virological response (SVR), which is associated with reduced occurrence of liver failure and HCC, and with prolonged overall survival (Backus et al., 2011, Cardoso et al., 2010 and Van der Meer et al., 2012). Many highly potential direct-acting antiviral (DAA) agents are being assessed in clinical trials and various combinations of DAA’s result in high SVR rates. Some DAAs target viral proteins, such as NS3/4A protease and NS5A/B replication inhibitors, whereas others target host factors that are essential for HCV replication, such as cyclophilin A or microRNA-122 (miR-122) (Flisiak et al., 2012 and Janssen et al., 2013). MicroRNAs (miRNAs) are small (19–24 nucleotides), non-coding, RNA molecules that are involved in various cellular processes by post-transcriptional suppression of gene expression (Ambros, 2004 and Bartel, 2004). MiR-122, a highly abundant miRNA expressed in the liver (Lagos-Quintana et al.

The weighted average CI was calculated using the formula: CI = [CI50 + 2CI75 + 3CI90 + 4CI95]/10, where CI50, CI75, CI90, and CI95 are the CI values at 50%, 75%, 90% and 95% inhibition, respectively ( Bassit et al., 2008 and Chou and Talalay, 1984). We assessed the effect of PYC on HCV in R6FLR-N and FLR3-1 cell lines after 72 h (Fig. 1). The data

are expressed as relative values using the relative light unit count for the 0 μg/mL treatment sample as 100% (Fig. 1A). The results showed that PYC inhibited luciferase activity in R6FLR-N cells (50% inhibitory concentration [IC50] = 5.78 ± 3.75 μg/mL, 50% effective concentration [EC50] = 4.33 μg/mL (2.2–8.5) in a dose-dependent Selleckchem Tofacitinib manner. To rule out the possibility that the antiviral activity was caused by cytotoxic effects, cell proliferation was analysed using the WST-8 assay; no significant differences in cell viability (50% cytotoxic concentration [CC50] > 60 μg/mL PYC; Selectivity index [SI] > 14.1) (Fig. 1B). These results http://www.selleckchem.com/products/cobimetinib-gdc-0973-rg7420.html indicate that PYC suppresses HCV (genotype 1b) replication. Consistent with results showing the inhibitory effects of PYC on HCV replication, we observed that HCV NS3 protein levels decreased significantly in PYC and IFN-alpha-treated HCV replicon cell lines (Fig. 1C). HCV NS3 and NS5B proteins levels were progressively

suppressed in HCV replicon cell lines at various PYC concentrations (0, 5, 10, and 20 μg/mL) (Fig. 1D). These results suggest that HCV protein synthesis was inhibited by PYC in a concentration-dependent manner. R6FLR-N cells were treated with IFN-alpha and RBV alone or in combination with several concentrations of PYC and incubated for 48 h (Fig. 2A). HCV replication was suppressed by approximately 20% following treatment with 5 μg/mL RBV, and by approximately 40% following treatment with 1 IU/mL IFN-alpha. Treatment with both RBV and IFN-alpha led to Sodium butyrate approximately

50% suppression. PYC showed a dose-dependent additive effect when administered in combination with RBV and IFN-alpha (Fig. 2A). Treatment with both PYC (5 μg/mL) and IFN-alpha (1 IU/mL) showed a synergistic effect (CI = 0.253) in suppressing HCV replication without cytotoxicity (Fig. 2A and B). JFH Luc3-13-N cells were inoculated with IFN-alpha (5 IU/mL) or several concentrations of PYC (5–50 μg/mL) and incubated for 72 h (Fig. 2C). HCV (genotype 2a) replication was suppressed by approximately 50% following treatment with 40 μg/mL PYC (Fig. 2C) without significant cytotoxicity (Fig. 2D). PYC, IFN-alpha, and RBV treatments were also evaluated in JFH-1/K4 HCV (genotype 2a) infected cells (Fig. 2E). HCV RNA levels decreased in the presence of PYC (10 or 20 μg/mL) to levels comparable to treatment with 1 IU/mL IFN-alpha in cell culture supernatant after 72 h.