Existing epidemiologic studies indicate that the use of

Existing epidemiologic studies indicate that the use of Bosutinib Src Swedish snus��a smokeless tobacco product low in TSNA��even though associated with an increased risk of pancreatic cancer when compared with never-users of any tobacco is not related to lung cancer and that the risk of oral cancer, if it exists, is very limited (Greer, 2011; Luo et al., 2007). Because of their potential for reducing exposure to TSNA and other carcinogens that are present in cigarette smoke, the use of low-TSNA smokeless products is seen by some as a potential harm-reduction strategy (Bates et al., 2003; Levy et al., 2004). Another critical chemical in smokeless tobacco is nicotine, the main known addictive constituent.

Both total nicotine and unprotonated nicotine content��the biologically available form of nicotine that depends on the pH of the product��are critical in consumers�� acceptance of a smokeless tobacco product and potential addiction to it (Fant, Henningfield, Nelson, & Pickworth, 1999; Henningfield, Fant, & Tomar, 1997). Initial analyses revealed that single pouches of Marlboro Snus, Camel Snus, and similar products contain relatively low levels of TSNA and nicotine as compared with conventional moist snuff, implying lower carcinogenic and addictive potential (Stepanov, Jensen, Hatsukami, & Hecht, 2008). However, information on chemical composition of these products is limited, and the extent of variability of TSNA and nicotine in a particular product because of manufacturing methods, the way the product is stored, or because of the intent of the manufacturer is unknown.

This information is essential in view of continuous modifications that this new category of products undergoes as it is being test marketed. Moreover, the potentially important role of TSNA and nicotine levels in labeling and marketing regulations, as well as in the establishment of standards for tobacco products as a part of Food and Drug Administration (FDA) regulation, makes the ongoing evaluation of these toxicants particularly important. The New Product Watch is a web-based national monitoring network made up mainly of state tobacco program staff and their community partners. It provides tools for monitors to periodically report local observations of new oral tobacco products being sold locally. Results are posted online for use by members around the country. Drug_discovery Twice a year, monitors collect product samples for analysis of chemical constituents and product packaging (Rogers, Biener, Nyman, & Crow, 2010). We present here the results of TSNA and nicotine analyses of Marlboro Snus, Camel Snus, and Camel Dissolvables that were purchased as part of the first phase of the New Product Watch project.

Table 2 Analytical methods Statistical analysis We followed the

Table 2. Analytical methods Statistical analysis We followed the definitions and procedures outlined in International Standardization Organization (ISO) documents (ISO 5725-1, ISO 5725-2, and ISO 5725-4) except SB203580 p38 MAPK inhibitor as indicated below. The first and last of these documents define trueness to describe the closeness of agreement between the arithmetic mean of a large number of test results and the true or accepted reference values (our targets for the pools in Table 1). Precision refers to the closeness of agreement between test results. An estimate of bias from an assessing laboratory is the difference between the mean of the pool target concentration and its accepted target reference value. To assess the significance of the bias estimates, we constructed 95% CIs around this difference, using equations from section 4.

7.2 in ISO 5725-4:1994(E). No significant bias was declared if the interval covered the value 0. Other concepts in the context of laboratory testing include repeatability, which characterizes variability among replicates obtained in the same laboratory on the same material (target concentration), and the between-laboratory variance, which refers to variability among laboratories using the same method. We used the approach outlined in ISO 5725-2:1994(E) to assess these characteristics (equations and, respectively). The sum of these two quantities is called the reproducibility variance (equation We also examined the functional relationship between precision values and the mean level for each of the target level samples, using ISO 5725-2:1994(E) following section 7.

5. It is not unusual for the repeatability (Sr) and reproducibility (SR) variances to follow a linear relationship with the mean values, often through the origin. We tested whether an intercept other than the origin was needed, and we excluded the intercept in the model if it was deemed nonsignificant. In examining precision according to ISO 5725-2:1994(E), we first obtained an overview of the data, using Mandel��s h and k charts before more detailed analyses. The statistic h is calculated for each participant laboratory and pool combination by a standardization of the mean of the replicate measurements for each laboratory/pool. The average of the replicates for all laboratories reporting on the pool is subtracted and the difference is divided by the standard deviation, again calculated over all participants for the given pool.

Note that h is either positive or negative when the laboratory obtains, on the average, a value higher (or lower) than the average of all laboratories for the given pool. A plot was made for all the h values for all pools, arranged by laboratories. The statistic k measures the relative variability between the GSK-3 replicate measurements of any laboratory for a given pool.

The HBEC mucosal surfaces were washed with 500 ��l PBS for 30 min

The HBEC mucosal surfaces were washed with 500 ��l PBS for 30 min prior to experimentation to remove selleckchem accumulated endogenous SPLUNC1. SPLUNC1 recovery into the ASL was measured over time by lavaging with 200 ��l PBS at timed intervals after the initial wash/volume load. To detect SPLUNC1, 20% by volume of the lavage was transferred onto a nitrocellulose membrane using a slot-blot apparatus (GE Healthcare, Pittsburgh, PA, USA). The blot was processed as described above using an anti-hPLUNC primary antibody (R&D Systems, Minneapolis, MN, USA). Transepithelial potential difference (Vt) studies A single-barreled Vt-sensing electrode was positioned in the ASL by micromanipulator and used in conjunction with a macroelectrode in the serosal solution to measure Vt using a voltmeter (World Precision Instruments, Sarasota, FL, USA), as described previously (16).

Circular dichroism (CD) G22-A39 (100 ��M) was dissolved in a buffer containing 10 mM sodium phosphate (pH 7.4) in a 1 mm cuvette. Using a Chirascan-plus instrument (Applied Photophysics Ltd., Leatherhead, UK), 5 individual spectra from 185 to 280 nm were recorded at 25 �� 1.0��C and averaged. All measurements were corrected for buffer signal. Expression and purification of human SPLUNC1 The SPLUNC1-��19 construct was created by cloning amino acid residues 20�C256 out of the SPLUNC1 cDNA described above for entry into pMCSG7 for protein expression. BL21-CodonPlus competent cells (Agilent Technologies, Santa Clara, CA, USA) were transformed with the expression plasmid and cultured in the presence of antifoam (50 ��l), ampicillin (100 ��g/ml), and chloramphenicol (34 ��g/ml) in Luria broth medium with vigorous shaking at 37��C until an OD600 of 0.

6 was attained. Expression was induced with the addition of isopropyl-1-thio-d-galactopyranoside (IPTG). Bacteria were collected by centrifugation at 4500 g for 20 min at 4��C. Cell pellets were resuspended in buffer A (20 mM potassium phosphate, pH 7.4; 50 mM imidazole; 500 mM NaCl; and 0.02% NaAzide) along with lysozyme, DNase1, and protease inhibitor tablets. Cells were sonicated, and cell lysate was separated into soluble and insoluble fractions using high-speed centrifugation. The soluble fraction was filtered, then flowed over a Ni-NTA His-Trap gravity column and washed with buffer A. The bound protein was eluted with buffer B (20 mM potassium phosphate, pH 7.

4; 500 mM imidazole; 500 mM NaCl; and 0.02% NaAzide) and separated using an S200 gel filtration column on an ?KTAxpress (GE Healthcare). The histidine tag was removed with Tev protease, leaving amino acid residues serine, GSK-3 asparagine, and alanine on the N terminus of the protein. Another round of nickel and HPLC purification removed the tag from the purified SPLUNC1-��19. We confirmed that SPLUNC1-��19 purified in this fashion inhibited ENaC HBECs (n=6).

Further, the primary source of maternal

Further, the primary source of maternal Tofacitinib CP-690550 SDP data comes from reports within 1 year of the child��s birth, addressing a limitation of long recall times discussed elsewhere (Kandel & Udry, 1999; O��Callaghan et al., 2006). Other exposure to tobacco smoke was not assessed which might occur from father or other family members smoking; such exposure may exacerbate the genetic, physiological, and social mechanisms of intergenerational transmission. We also did not account for other methods of nicotine use, such as smokeless tobacco or smoking of tobacco through means other than cigarettes (e.g., pipe, cigar). These will be likely to bias our results in the direction of the null hypothesis as children who were exposed to nicotine will have been classified incorrectly as nonsmokers.

Some issues arise with the sample. The sample was large, but even such a large sample led to relatively small numbers of individuals in, for example, the early experiment group with a sample of 127. This small sample size has limited statistical power in analyses including this group. The data on smoking are recorded biennially; therefore, our trajectories cannot capture short-term fluctuations in cigarette behavior but rather describe overarching patterns of youth smoking across adolescence and young adulthood. Finally, due to the cohort structure of the NLSY79-CYA sample, the most complete portion of data covers the period age 14�C16 and thus may be less representative of smoking patterns in early adulthood than have been described in previous population representative studies The small size of the mediation effect, relative to the direct effect of maternal smoking (of any class) means that the result is vulnerable to omission of control variables that are precursors to maternal smoking, child smoking, and child behavior problems.

Although we included a wide range of control variables, additional unmeasured covariates that are associated with all three variables of interest would create an artifactual mediation effect. The results of this study provide a rigorous test of the mediation through behavior problems hypothesis and suggest that the mechanism of intergenerational transmission of smoking via SDP follows a more direct route, potentially via a physiological mechanism. However, maternal smoking after Entinostat pregnancy seemed to be associated with youth smoking, at least in part, because of a mediation effect of problem behavior.

The move to print media allowed the industry to rely increasingly

The move to print media allowed the industry to rely increasingly on images rather than words (King et al., 1991). Companies purchased full- or double-page advertisements, prominently placed on the right-side pages and back covers of magazines (Weinberger et al., 1981). The industry also redirected resources to sponsorship marketing. Despite the broadcast ban, companies Vorinostat chemical structure received brand exposure on television by sponsoring sports events such as auto racing (Blum, 1991; Siegel, 2001; Zwarun, 2006). Then, in 1998, the Master Settlement Agreement (MSA) between the attorneys general of 46U.S. states and the 5 largest tobacco companies banned cigarette billboard advertising in the United States. Again the tobacco industry redirected resources by increasing the amount of POS advertising.

This included greater exterior cigarette advertising at tobacco retail locations (e.g., on the windows and doors of gas station mini-marts), as well as greater interior POS promotions (e.g., interior advertisements, sales promotions such as multipack discount offers, branded objects such as clocks and shopping baskets) (Celebucki & Diskin, 2002; Wakefield et al., 2002). Other countries have seen a similar shift toward POS advertising, as POS remains the least regulated channel for tobacco marketing (Henriksen, 2012). The industry also has responded to stronger legislation by redirecting marketing resources to other geographic areas within a given country.

When China passed the Control of Tobacco Products law in 1992��which banned print and broadcast media advertising��BAT increased communication spending by 43% to take advantage of advertising opportunities in provinces that had not yet fully embraced the ban (Lee, Gilmore, & Collin, 2004). Specifically, BAT exploited regionality, recognizing that local attitudes toward the national ban might be different than those in Beijing, and they capitalized on provinces�� need for foreign currency. The industry also realized that local interpretations of the legislation could allow Dacomitinib for greater flexibility and, in turn, relaxed restrictions (Lee et al., 2004). Last, in countries with comprehensive or otherwise strict bans, the tobacco industry has used several tactics to combat regulation. One strategy has been to use cross-border advertising to their advantage. In Singapore, where strict tobacco marketing laws have been in place since 1971, the tobacco industry used the absence of regulations in nearby Malaysia to advertise tobacco to Singaporeans: both BAT and PM exploited the ��spillover�� of Malaysian television into Singapore (Assunta & Chapman, 2004c). Another industry tactic is challenging marketing restrictions in court (e.g., citing violations of free speech).

05) Mapping the Forms of the Significant Interactions Based on t

05). Mapping the Forms of the Significant Interactions Based on the recommendations of Cohen and Cohen (1983, p. 323), the forms of the significant interactions were examined by inserting specific values for each predictor variable into the equations associated with the described analysis (half SD above and below the mean for PDS total). As depicted in Figure 1, the significant interactive selleck Imatinib effects are evident at Minutes 3 (p = .06) and 4 (p < .05) of the challenge paradigm only. Inconsistent with prediction, the highest levels of anxiety ratings were reported by the smoking-as-usual groups. Discussion Consistent with prediction, posttraumatic stress symptom severity was significantly predictive of peri-challenge anxiety.

Specifically, posttraumatic stress symptom severity was incrementally predictive of anxious responding at Minutes 3 and 4 of the challenge, though it demonstrated only a trend toward statistical significance at Minutes 1 and 2 of the challenge (p��s = .07). This finding is consistent with past work (Feldner et al., 2008). These results utilize a trauma-exposed sample with varying levels of posttraumatic stress (mostly without PTSD) to extend previous work, which demonstrated that smokers with PTSD compared with those without PTSD report more anxiety and smoking craving following both trauma-related and general stress scripts (Beckham et al., 2007). Furthermore, the observed difference with respect to the role of posttraumatic stress symptom severity and anxious arousal at Minutes 1�C2, and in contrast to Minutes 3�C4 of the challenge, may be due to an increasing perceived intensity of the challenge over time as well as limited statistical power as a function of the sample size.

Contrary to prediction, the main effect of 12-hr cigarette deprivation was not a significant predictor of peri-challenge anxious responding. The smoking-as-usual group (in contrast to cigarette deprivation) emerged as a significant incremental predictor of anxious responding at Minutes 1, 3, and 4 of the challenge paradigm. This finding is consistent with the cigarette deprivation effects noted in at least one previous study using the identical laboratory paradigm (Vujanovic & Zvolensky, Brefeldin_A 2009). Furthermore, this unexpected finding might be attributable to at least three possible nonmutually exclusive reasons. First, despite random assignment of participants to the cigarette deprivation versus smoking-as-usual groups, the latter group evidenced a higher total number of Axis I diagnoses (23 total diagnoses among 36 individuals) as compared with the cigarette deprivation group (13 total diagnoses among 27 individuals).

falciparum; using msp2 PCR, 97/388

falciparum; using msp2 PCR, 97/388 www.selleckchem.com/products/dorsomorphin-2hcl.html individuals were found positive, giving a prevalence rate of 25% P. falciparum malaria. RFLP analysis was successfully conducted on 80 samples. The msp2 PCR fragment revealed 56 (70%) single infections, 18 (22.5%) double infections, five (6.25%) triple infections and one (1.25%) quadruple infection. The mean multiplicity of infection (MOI; mean number of co-infecting parasite clones) in P. falciparum positive samples was 1.39. Ten PCR reactions were performed for the five genes associated with drug resistance. Of the 97 P. falciparum positive samples analysed, 21 (21.6%) showed poor amplification for the majority of the PCR reactions (> 5 poor PCR results out of 10) and had thus to be excluded. The 76 (78.4%) remaining samples were used for genotyping by microarray.

Sixteen unclear microarray typing outcomes were confirmed by sequencing of the corresponding gene fragments (Macrogen Inc., Korea). Overall, the analysed population showed to be very homogenous (Figure (Figure2).2). Two different haplotypes were observed for the two CQ resistance-associated genes. With 70% of the patients having a single infection and the unambiguous DNA microarray signals, it was considered that co-infecting clones had the same pattern of SNPs. The most dominant haplotype showed mutations at positions pfmdr1 N86Y and pfcrt C72 S, N75D/E, K76T, A220 S, N326 D, and I356L, and was observed in 98.4% (61/62) of the population. The second haplotype was observed in one sample only (1.6%) and differed from the first one by having one additional mutation at position N1042 D in pfmdr1.

Other SNPs related to CQ resistance were all wild-type. Figure 2 Observed haplotypes of SNPs related to resistance to CQ, SP and artemisinin derivatives. A haplotype is defined by the combination of mutations in genes related to drug resistance. Grey indicates mutations and white indicates wild-type. Two different … For the genes involved in SP resistance, mutations were fixed at positions C59R and S108N in pfdhfr. Other SNPs in pfdhfr and pfdhps were all wild-type. No mutation was detected in pfATPase6. Discussion The outcome of CQ+SP against falciparum malaria was investigated at a clinic located close to Honiara in the SI. At the same time, the presence of resistance-associated SNPs in P. falciparum was assessed in the surrounding asymptomatic community, screened by a cross-sectional survey.

The resistance-associated SNPs observed in the sample set are in line with previous findings from other malaria areas worldwide. Regarding SP resistance, all samples presented two fixed mutations at positions C59R and S108N in pfdhfr and none in pfdhps. The importance of cumulative mutations conferring SP resistance has been described; in particular the combination Cilengitide of the SNPs pfdhfr S108N, N51I, C59R and pfdhps A437G, K540E [9,28] being correlated with anti-folate treatment failure.

MATERIALS AND METHODS Rat Experiment Animals Twelve female Sprag

MATERIALS AND METHODS Rat Experiment Animals. Twelve female Sprague-Dawley rats (Harlan Laboratories, Venray, The Netherlands), weighing 250�C300 g, were used. The rats were acclimatized to the animal facility for at least 1 wk. Rats were housed in groups of five in an enriched environment with free access http://www.selleckchem.com/products/Bicalutamide(Casodex).html to food (standard pellets, R3, Lactamin, Kimstad, Sweden) and water on a 12:12-h light-dark cycle. The estrous stage of the rats was not accounted for in the present study. All experiments were approved by the local animal ethics review committee in G?teborg, Sweden (403-2008) and conducted at AstraZeneca, M?lndal, Sweden. Surgery. Prior to surgery, rats were administered antibiotics (1 ml/kg) (Bactrim, Oral 40/8 mg/ml, Roche, Basel, Switzerland) and analgesic (10 mg/kg) (Romefen, Merial Norden, Skovlunde, Denmark).

Rats were anesthetized with isoflurane (2�C3 vol%) (Forene, Abbott Scandinavia, Solna, Sweden) and kept on a heating pad during surgery to maintain body temperature. Surgical procedures consisted of implantation of skull electrodes and positioning of an abdominal connector. A midline scalp incision was made and the abdominal cavity was exposed by a 5-cm midline incision. From the abdominal cavity, Teflon-coated stainless steel cables (? = 0.3 mm; Cooner Wire, Chatsworth, CA) were passed through the abdominal musculature and tunneled subcutaneously across the thorax to the incision on top of the head. Two centimeters lateral to the abdominal incision a small plastic connector (AstraZeneca) was exteriorized and sutured to the abdominal wall to allow future access to the electrode cables.

The abdominal incision was closed and the rats were then placed in a stereotaxic frame. A few drops of lidocaine (Xylocaine iv, 20 mg/ml, AstraZeneca, S?dert?lje, Sweden) were applied to the scalp incision and connective tissue was scraped away from the skull surface. After exposure of the skull, a small hole (?0.9 mm) was drilled on each of the electrode positions and cables were secured by titanium electrodes (?0.9 mm, Wennbergs finmek, Gunnilse, Sweden) into these positions. Three electrodes for monopolar recordings were placed on the right side of the skull (Fig. 1A); the positions were selected based on experience from a previous study (23). Briefly, three electrodes were placed 1.5 mm lateral to the sagittal line. The most anterior electrode was placed 1.

5 mm anterior to bregma and the following two electrodes in a rostrocaudal direction, separated by 3 mm (Fig. 1A). The reference electrode was positioned 2 mm caudal to lambda. Fig. 1. A: positioning of recording electrodes Brefeldin_A on the rat skull. B: schematic drawing of the experimental protocol used in the rat study. C: schematic drawing of the experimental protocol used in the human experiment. Electrodes were placed avoiding penetration of the underlying dura mater.

Briefly, fresh kidney cortical samples were homogenized

Briefly, fresh kidney cortical samples were homogenized Leukemia as above in ice-cold isolation buffer containing fresh protease inhibitors, and crude membranes were obtained by centrifugation at 48,000 g for 1 h at 2��C (Beckman J2�C21M; JA-20 rotor; Beckman Coulter, Fullerton, CA). Pellets were resuspended, homogenized in a Dounce glass homogenizer, and subjected to Mg2+ aggregation by addition of MgCl2 to a final concentration of 15 mM at 4��C for 20 min. Aggregated membranes were removed by centrifugation at 3,000 g for 10 min at 2��C (Beckman Allegra 21R; Beckman Coulter), and the supernatant was subjected to two additional rounds of Mg2+ aggregation as above (pellets discarded), followed by centrifugation at 48,000 g for 30 min at 2��C. The resulting pellet enriched in BBMV was used for Na+/H+ exchange activity experiments and for immunoblotting.

Na+/H+ exchange activity in BBMV. Na+/H+ exchange activity was measured in fresh BBMV with the acridine orange fluorescence quenching method (53) as previously described (6). Briefly, the BBMV pellet obtained as above was rehomogenized in intravesicular solution [280 mM mannitol, 5 mM 2-(N-morpholino)ethanesulfonic acid (MES), pH 5.5] with a Potter-Elvehjem Teflon-glass homogenizer, equilibrated for 2 h at 4��C, centrifuged at 48,000 g, and resuspended with a 27-gauge syringe needle and intravesicular solution to a final protein concentration of 10 mg/ml. Extravesicular solution [120 mM N-methyl-d-glucamine (NMDG)-gluconate, 20 mM HEPES, pH 7.5] with fresh acridine orange (Invitrogen) added to a final concentration of 6 ��M was loaded in a cuvette equipped with a magnetic mini stir bar.

Acridine orange fluorescence was monitored in a computer-controlled spectrofluorometer (��excitation = 493 nm, ��emission = 530 nm; QM-8/2003, Photon Technology International, London, ON, Canada). Fluorescence was rapidly quenched by addition of acid-loaded BBMV (a volume containing 150 ��g protein) in the absence of Na+ and under constant stirring. Addition of extravesicular Na+ (Na+-gluconate to a final concentration of 30 mM) activated Na+/H+ exchange and resulted in a proportional increase in fluorescence. Replacement of Na+-gluconate with NMDG-gluconate was used to monitor Na+-independent quenching. Na+/H+ exchange activity was estimated as the initial rate of increase in fluorescence on addition of Na+-gluconate, from which the fluorescence increase with NMDG-gluconate (typically <1/10th of that with Na+-gluconate) was subtracted.

Quantitative comparisons were made between BBMV prepared at the same time from Anacetrapib rats killed on the same day. NHE3 antigen in BBMV. Rat kidney cortex BBMV prepared as above were homogenized in RIPA buffer (150 mM NaCl, 50 mM Tris?HCl, pH 7.4, 5 mM EDTA, 1% Triton X-100, 0.5% deoxycholate, and 0.1% SDS) containing fresh protease inhibitors and cleared by centrifugation (14,000 g, 4��C, 30 min), and protein content was determined by the method of Bradford.

Serum HER2 levels above 1000 ng/mL was an indicator that standard

Serum HER2 levels above 1000 ng/mL was an indicator that standard doses of Trastuzumab may be insufficient as reflected by low concentrations of serum Trastuzumab.53 Several studies now support the observation that a significant decrease in sHER2 levels from baseline to 30�C90 days after initiation of treatment is a predictor selleck kinase inhibitor of outcome to HER2-targeted therapies. In contrast, patients with increasing sHER2 levels, a persistently high sHER2 level or who do not achieve at least a 20% decline in sHER2 levels in the early weeks and months of HER2 targeted therapies may benefit from treatment with new HER2 inhibitors that are in clinical development.52,54 Since patients can have increases in sHER2 levels that occur earlier than actual clinical signs of recurrence, routine monitoring of sHER2 levels can be an early warning sign for physicians and patients to consider additive or alternative therapy strategies.

Earlier detection of cancer progression is an actionable event for the oncologist and one which can trigger intervention with the variety of therapies or combination of therapies that are now available for breast cancer patients. In theory, the sooner one attacks a tumor, particularly while the tumor burden is low, the better the probability of a positive patient outcome. Well-defined clinical studies are needed to test this hypothesis. Studies conducted thus far have collectively shown that patients treated with hormone therapy, chemotherapy, Trastuzumab, Lapatinib, or a combination of these therapies have serial changes in sHER2 levels that paralleled changes in the clinical course of disease; therefore, patients with HER2-positive breast cancer can greatly benefit from routine monitoring of thes HER2 level.

14�C17,21,22,25,52,54 Controversy Surrounding Serum HER2 Testing in all Breast Cancer Patients as a Prognostic Biomarker In contrast to the large number of publications that illustrate the clinical value of sHER2 testing, a handful of publications have reached negative conclusions regarding the clinical value of measuring this specific circulating biomarker for HER2-positive breast cancer. Articles by Lennon et al55 Leary et al56 and Leyland-Jones et al57 are the most notable. All three publications agreed with the 2007 ASCO guidelines and did not recommend sHER2 testing for clinical use.

The articles by Leary et al and Leyland-Jones et al primarily reviewed publications that used research use only assays that were not standardized or validated by acceptable diagnostic protocols and procedures. In general, the research use only assays listed in the data tables of these publications did not define the specificity of the capture and detection antibodies in the assay nor was there a definition of what constitutes a statistically significant change between 2 serial blood Anacetrapib samples.