ntr.oxfordjournals.org Funding This work was supported by the NCI at the National Institutes of Health (sponsor award number HHSN261200900395P) Ruxolitinib CAS awarded to JNS. Declaration of Interests None declared. Supplementary Material Supplementary Data: Click here to view. Acknowledgments The authors would like to thank the Deputy Editor and both reviewers for their valuable feedback that helped improve the manuscript, senior program analyst James Gibson for providing the data set and science writer Anne Brown Rodgers for her editorial suggestions.
Eight EC users (3 women; 8 Caucasian) consented to and completed this single-session (5 hr) Institutional Review Board-approved protocol. Participants used their own fully charged EC, and a prefilled cartomizer (heater + liquid) of their chosen flavor/nicotine content was provided.

At the start of the session, participants were surveyed regarding health, demographics, and EC use habits. Participants were included in the study if they reported using an EC for at least 3 months, used at least 2�C3 ml of nicotine solution or two cartridges per day, used nicotine solution of at least 10 mg/ml nicotine, smoked fewer than five cigarettes per day, and were between the ages of 18 and 55. Participants were excluded if they reported any chronic health or psychiatric conditions, had a history of high or low blood pressure, or were unwilling to use a cartridge or cartomizer during the session. Participants were required to abstain from all nicotine and tobacco product use, including ECs, for at least 12 hr prior to the session.

Abstinence was verified upon arrival at the laboratory by a carbon monoxide level of 10 ppm or less and was further verified later by plasma nicotine levels at or below the limit of quantification (2 ng/ml). At session onset, a venous catheter was inserted into the participant��s forearm vein and continuous physiological monitoring commenced. Approximately 55 min later, blood was sampled, and 5 min after that, participants were provided with their EC and were instructed to take 10 puffs (30-s interpuff interval [IPI; as in Eissenberg, 2010; Vansickel et al., 2010]). Blood was sampled 5 and 15 min after the start of the 10-puff period. Then, a 60-min ad lib puffing period began, and blood was sampled every 15 min. Participants then entered a 2-hr rest period during which no puffing occurred, and blood was sampled every 30 min.

Participants completed subjective questionnaires at baseline and again after the 10-puff, ad lib, and rest periods. At session��s end, the venous catheter was removed, physiological monitoring ceased, and participants were compensated. Physiological outcome measures included heart rate (measured every Entinostat 20 s) and plasma nicotine concentration (blood centrifuged and plasma stored at ?70 ��C for analysis of nicotine concentration).

Note that the ordering of classes is somewhat arbitrary, such that different permutations of the same four classes can result from different starting values. In this analysis, it www.selleckchem.com/products/tofacitinib-cp-690550.html was apparent that the ordering of the classes was not consistent across the datasets (e.g., the nonsmoking classes might be the first class for some imputed datasets and the second class for others), consequently we chose to carry out the LCA on each dataset in turn and pool the results ourselves. Regression Modeling of Risk Factors To examine the relationship between risk factors and our latent classes, a two-stage modeling procedure was used. Following the LCA, class assignment probabilities were exported to in Stata version 11-MP2 (StataCorp., 2009).

These probabilities were incorporated as an importance weighting (iweight) in a series of univariable multinomial logistic regression models. For the imputed data, the 100 imputed datasets were re-stacked and analyzed using by Stata��s mi routines. Results Characteristics of Participants The starting sample for these analyses is the 13,973 singletons/twins who survived at least until 1 year of age. Of these, 3,038 respondents provided complete data on the three measures (1,245/41% boys and 1,793/59% girls). A further 2,232 missed one response and 2,052 missed two responses, resulting in a dataset of 7,322 with at least one of the three measures (3,351/46% boys and 3,971/54% girls). First, the relationship between level of response (complete/partial/none) and baseline demographics was studied.

There was evidence of an association between level of response and gender as well as housing tenure, parity, and maternal education with complete responders being more likely to be female, have parents who own their own home, have a mother educated beyond high school, and have fewer siblings (data available on request). Level of response was also related to smoking frequency. Table 1 shows the relationship between smoking frequency at each time point and whether the respondent provided complete or partial smoking information. There is strong evidence that those who provide partial smoking information are more likely to smoke regularly. Table 1. Relationship Between Smoking Frequency and Degree of Response Latent Class Analyses Nonimputed Data For the latent class model, there were 64 possible patterns of response.

Of these, 46 were observed in the complete case dataset of 3,038 participants. This sample was dominated by a nonsmoking GSK-3 pattern (none/none/none), which was the response for a total of 2,339 (77%) respondents. There was little evidence of regular smoking before the age of 15, with other common response patterns being none/none/occasional (n = 151, 5.0%), none/occasional/none (85, 2.8%), and none/none/weekly (52, 1.7%). The partially missing dataset contained an additional 57 response patterns.

To confirm CRNN expression in xenograft tumor, IHC staining with www.selleckchem.com/products/AZD2281(Olaparib).html anti-CRNN antibody was performed in sections (5��m in thick) of paraffin-embedded xenograft tumor. RNA interfering (RNAi) CRNN-expressing clones CRNN-C2, C3 (KYSE30) or CRNN-C1 (KYSE180) were transfected with double-stranded siRNAs (Ambion, Carlsbad, CA) with lipofectamine 2000TM reagent (Invitrogen) according to the manufacturer��s instructions. Forty-eight hours after transfection, the gene-silencing effect was measured by qRT-PCR and western blot analysis, respectively. Three independent experiments were performed. Cell cycle analysis CRNN-C2/CRNN-C1 or Vec-30/Vec-180(2��105) were fixed in 70% ethanol and stained with propidium iodide, and DNA content was analyzed by Cytomics FC (Beckman Coulter, Indianapolis, IN).

Western blot analysis Western blotting was done according to the standard protocol with antibodies for GAPDH, CRNN (Santa Cruz Biotechnology, Santa Cruz, CA), P53, P21WAF1/CIP1, cyclin D1, CDK4, cyclin E, CDK2, Rb, tubulin (Cell Signaling Technology, Danvers, MA). Statistical analysis Statistical analysis was performed using SPSS standard version 13.0 software (SPSS Inc, Chicago, IL). Data were expressed as mean �� S.E.M. from at least three independent determinations. Significance of difference was analyzed using Student��s t-tests. The correlation between CRNN expression and clinicopathologic characteristics was analyzed using the Fisher��s exact test. Cum survival was calculated from the date of diagnosis to the date of cancer-related death or last follow-up.

Survival curve was assessed by the Kaplan-Meier method and compared by the log-rank test. Relative risks of cancer-related death associated with CRNN expression status and other predictor variables were estimated by univariate analysis. Multivariate survival analysis was done on all parameters that were found to be significant on univariate level using the Cox regression model. Differences were considered significant for P<0.05. Results Downregulation of CRNN is frequently detected in ESCC The mRNA expression of CRNN in 9 ESCC cell lines and 56 primary ESCC tumors and their paired non-tumorous tissues were detected by semiquantitative and qRT-PCR, respectively. The results showed that downregulation of CRNN was detected in 26/56 (46.4%) of primary ESCCs (Figure 1A) and 9/9 of ESCC cell lines (Figure 1B).

Carfilzomib Figure 1 Downregulation of CRNN in ESCC. CRNN downregulation correlates with poor outcome of ESCC To investigate the clinical significance of CRNN downregulation in esophageal carcinogenesis, CRNN expression in protein level was also studied using ESCC tissue microarray. Expression of CRNN was classified into absent (scored as 0), weak-positive (scored as 1+) and strong-positive (scored as 2+) cytoplasmic staining. Informative expression of CRNN was detected in 249 ESCC cases.

All analyses were performed with SAS software, version 6.12. RESULTS A total of 154 patients participated in the study from January 2003 to June 2007. No patient withdrew from the trial during 6 mo follow-up. Seventy-one subjects were assigned in group A, whose average tumor size was 4.3 �� selleckchem 1.1 cm. Eighty-three participants were in group B, whose average tumor size was 4.1 �� 1.0 cm. The characteristics of the two groups are shown in Table Table1.1. There was no significant difference between the two groups in patients�� clinical profile except for the gender ratio. Table 1 Pre-RFA clinical profile of patients (M0N0) n (%) For patients in group A, the feeding artery was blocked in 66/75 (88%) HCC lesions, and decreased in size in 9/75 (12%) lesions. The average number of punctures per HCC was 2.

76 �� 1.12. Complete necrosis rate at 1 mo after RFA was 90.67% (68/75), and recurrence rate at 6 mo was 17.33% (13/75, lesions) (Figure (Figure2).2). For patients in group B, the average number of punctures per HCC was 3.36 �� 1.60. Complete necrosis rate at 1 mo after RFA was 90.20% (92/102), and recurrence rate at 6 mo was 31.37% (32/102). Figure 2 A 65-year-old man with cirrhosis and Child-Pugh class A liver function. An HCC lesion was diagnosed during regular US examination. A: CT showed a tumor with a size of 3.2 cm �� 3.0 cm in the right liver lobe; B: US showed a tumor (arrow) of 4.3 … No significant difference was found in the necrosis rate at 1 mo post-RFA. However, there was a significant different between the two groups in the average number of punctures per HCC and recurrence rate at 6 mo after RFA.

The recurrence time between the two groups was also significantly different by the log-rank test (��2 = 5.23, P = 0.02; Figure Figure3).3). When adjusted for gender, age and cirrhosis, it was still significantly different (��2 = 4.58, P = 0.03). Figure 3 The Kaplan-Meier curves for 6-mo recurrence rate in the two groups. In group A, RFA-related major complications were seen in five patients (7.04%), including three cases of pleural fluid collection, one of bowel wall edema, and one of intrahepatic biliary duct dilation with jaundice, which were all relieved by conservative therapy. A small amount of subcapsular hemorrhage around the puncture site, approximately 0.5-1 cm thick, during RFA was seen with US in 13 patients (18.31%) (Figure (Figure4).

4). It was not considered as a major complication because homeostasis was achieved after injection AV-951 of hemocoagulase, without additional intervention and no change in blood pressure. In group B, major complications were detected in nine cases (10.84%), including one of pneumothorax with pleural fluid collection, six of pleural effusion, one of abdominal wall abscess, and one of intraperitoneal hemorrhage in a tumor > 5 cm and protruding liver lateral surface. All complications were controlled with conservative treatment.

Absorbance was measured at 570nm using a Multiskan MS ELISA plate reader (Thermo Fisher Scientific Inc., Waltham, MA, USA). Laser microdissection Samples were derived selleck chem Rucaparib from surgically removed tissue from six patients with moderately differentiated, Dukes B stage, left-side CRC. In parallel, six adenoma specimens were collected. Paired control non-tumour tissues from patients were obtained from a clinically unaffected site near the resection end and were histologically normal. Tissue samples were immediately frozen in liquid nitrogen after surgery and were stored at ?80��C until the cutting period. Frozen tissue was placed in a cryomold with Tissue Tek embedding medium on dry ice for 1min. Frozen tissue specimens were cut in a series of 6-��m-thick sections onto PALM membrane-mounted glass slides at ?20��C.

After cutting, the slides were taken into dry ice, and were stored at ?80��C until microdissection for up to 48h before staining and dissection. The frozen sections were fixed in ethanol series, and were stained using cresyl violet (Sigma-Aldrich). After staining the tissue, tumour and normal tissues were diagnosed by the pathologist. A total of 5000 epithelial cells were collected from each section using the PALM system (PALM, Bernried, Germany). Microarray analysis Total RNA was extracted from HT29 cells using the RNeasy Mini Kit (Qiagen Inc., Germantown, MD, USA) and from LCM cells using the RNeasy Micro Kit (Qiagen Inc.), according to the manufacturer’s instructions. The quantity and quality of isolated RNA were tested by measuring absorbance and capillary gel electrophoresis using the 2100Bioanalyzer and RNA 6000 Pico Kit (Agilent Inc.

, Santa Clara, CA, USA). Biotinylated cRNA probes were synthesised from 1 to 5��g total RNA and fragmented using the One-Cycle Target Labeling and Control Kit (http://www.affymetrix.com/support/downloads/manuals/expression_s2_manual.pdf), according to the Affymetrix description. In case of LCM samples, two-cycle T7-based linear amplification was performed according to instructions of the manufacturer (Affymetrix Inc., Santa Clara, CA, USA). A volume of 10��g of each fragmented cRNA sample was hybridised into HGU133 Plus2.0 array (Affymetrix) at 45��C for 16h. Slides were washed and stained using Fluidics Station 450 and an antibody amplification staining method according to the manufacturer’s instructions.

Fluorescent signals were detected by a GeneChip Scanner 3000 (Affymetrix). Fifty-three microarrays from colonic biopsy samples (11 normal, 20 villous adenoma, 22 CRC) had been hybridised earlier, their data files were used in a previously published study using different comparisons (Galamb et al, 2008a,2008b, 2009) and are available in the Gene Expession Drug_discovery Omnibus database (series accession numbers: “type”:”entrez-geo”,”attrs”:”extlink”:”1″,”term_id”:”4183″,”text”:”GSE4183″GSE4183 and “type”:”entrez-geo”,”attrs”:”extlink”:”1″,”term_id”:”10714″,”text”:”GSE10714″GSE10714).

22 Witten et al16 suggested that exposure to petrol or diesel engine exhausts may increase the risk of lung cancer and neurological conditions in rats. A recent study in the US showed that breathing air polluted by exhaust fumes was responsible for more than 70% of the cancer risk in the South Coast Air Basin in California.41 selleck inhibitor Lopez-Abente et al42 correlated gastric cancer risk to consumption of a local wine sealed with a tar like substance obtained through boiling and distilling fir and pinewood which contains PAHs. Sinha et al43 associated increased risk of colorectal adenomas with benzo(a)pyrene intake in food. Tobacco smoke, which contains PAHs, has been implicated in the lung cancer.44 An association between PAH-DNA adducts and breast cancer incidences have also been reported.

18,19,36,45 Furthermore, there was a positive correlation regarding the distance to source of pollution and the level of 1-hydroxypyrene in blood samples of exposed animals. 1-hydroxypyrene level was significantly (P < 0.05) increased in groups of rats that had experienced dermal and inhaled exposure to the generator fumes compared to the group of unexposed rats (control). This points to the fact that the closer the rat to the generator exhaust source, the higher the absorption of these carcinogens after inhalation and dermal contact. Reports have suggested that most PAHs are well absorbed in mammals.30,46 Rapid absorption has been recorded in rats exposed to benzo(a)pyrene through inhalation.46 There is a high tendency of malignant tumor development in these PAH poisoned rats due to mutation arising from PAH-DNA adducts disrupting normal DNA transcription, translation, and replication.

However, gene polymorphisms in most enzymes have been identified in human beings and this could modulate individual cancer susceptibility. Ueng et al47 reported that exposure of rats to motorcycle exhaust and organic extracts of the exhaust particulate causes a dose- and time-dependent increase in cytochrome P-450-dependent monooxygenases as well as glutathione-S-transferase in the liver, kidney, and lung microsomes. This occurs as these enzymes metabolize the PAHs to polar nucleophilic metabolites that bind with the adenine and guanine bases of the DNA.47 Lin et al28 reported that the cytotoxicity of traffic related nano/ultrafine particle extracts was significantly higher than for coarser particles. This would be most likely due to the higher PAH concentration Carfilzomib in the fine generator exhaust particles. Conclusion The data available from this study shows that generator fumes contribute significantly to the atmospheric level of PAHs and that the level is dependent on the distance from the point of generation.

Cell viability/TNF cytotoxicity assay. YAMC, COX-1?/? MCE, and COX-2?/? MCE cells were plated in growth medium without interferon-�� and incubated for 2 days at 33��C. The cells were then starved in serum-free sellectchem RPMI at 37��C and treated with 100 ng/ml TNF for 48 or 72 h at 37��C. Cell number was determined using the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) reagent from the CellTiter 96 Aqueous One Solution cell viability assay (Promega, Madison, WI) or the NucleoCounter (New Brunswick, Edison, NJ) automated cell counter according to the manufacturers’ instructions. Relative cytotoxicities are expressed as percentage of cell loss stimulated by TNF, calculated as follows: 100% ? A490(TNF)/A490(Mock) �� 100%, where A490 is absorbance at 490 nm.

Generation of stable cell lines. The generation of EGFR?/? MCE cells with empty pcDNA3.1/Zeo vector (Vec) or wild-type (WT) EGFR and TNFR1?/? MCE cells with empty bicistronic LZRS-green fluorescent protein retroviral vector (Vec) or hemagglutinin (HA)-tagged WT mouse TNFR1 is described elsewhere (16, 76). An HA-tagged mouse death domain deletion (��DD) TNFR1 mutant, in which the TNFR1 protein sequence ends at P348, and HA-tagged mouse WT TNFR2 were cloned from YAMC cells. The resulting amplified sequences were subcloned into the LZRS vector. TNFR1?/? and TNFR2?/? MCE cells were infected with the respective retroviral constructs selected for expression, as previously described (16). Small interfering RNA transfections.

YAMC cells were transfected with 200 nM siGenome SMARTpool number 2 nontargeting (NT) small interfering RNA (siRNA) or 200 nM mouse EGFR siGenome SMARTpool siRNA (Dharmacon, Lafayette, CO) using the Lipofectamine RNAiMax (Invitrogen, Carlsbad, CA) transfection reagent according to the manufacturer’s instructions. Cell stimulation assays, preparation of cell lysates, and Western blot analysis. Cells were treated with 100 ng/ml TNF or 10 ng/ml EGF (Peprotech, Rocky Hill, NJ), unless otherwise indicated. For experiments using pharmacological inhibitors, cells were pretreated for 1 h with DMSO vehicle control or 1 ��M AG-1478 (Calbiochem, San Diego, CA), 2 ��M CGP-77675 (Pharma Research, Basel, Switzerland), 1 ��M PP2 (Calbiochem), 10 ��M SB-203580 (Calbiochem), or 10 ��M SB-202190 (Calbiochem).

Primary and secondary antibodies used in Western blot analysis include rabbit anti-EGFR (Millipore, Billerica, MA), mouse anti-actin (Sigma-Aldrich, Dacomitinib St. Louis, MO), rabbit anti-HA (Zymed), horseradish peroxidase-conjugated anti-mouse and anti-rabbit polyclonal antibodies (Cell Signaling Technology, Beverly, MA), and goat anti-COX-2 and horseradish peroxidase-conjugated anti-goat polyclonal antibodies (Santa Cruz Biotechnologies, Santa Cruz, CA).

Filled circles represent a significant change from the 0.05-mg nicotine yield postcigarette measurement, while asterisks (*) … Visual selleckchem Rapamycin Analog Scale. Nicotine effects on VAS stimulated varied as a function of sensation-seeking status (Figure 2, Panel B). Simple effects indicated that increases in ratings occurred as a function of nicotine yield in low sensation seekers alone, while high sensation seekers reported higher ratings of stimulated than low sensation seekers after smoking the 0.05-mg cigarette (p = .07). These results were similar to those on MNWS restlessness, with changes in ratings among high sensation seekers dependent on smoking but independent of nicotine yield, and changes among low sensation seekers dependent on nicotine yield.

Nicotine effects on other VAS items did not differ as a function of sensation-seeking status. Panel B of Figure 3 presents the pooled effects of nicotine yield on ratings of anxious. Significant effects of nicotine yield were found, with follow-up testing indicating reductions in deprivation-induced ratings of anxious at 0.6- and 0.9-mg yields relative to 0.05 mg. Panel C of Figure 3 presents a main effect of nicotine yield on ratings of light headed. Follow-up testing indicated increases in ratings of light-headed at the 0.6- and 0.9-mg yields relative to 0.05 mg. Similar effects were found on ratings of head rush (Figure 3, Panel D), stimulated, like drug, pleasant feeling, and sedated. Profile of Mood States. Nicotine effects on POMS elation varied as a function of sensation-seeking status (Figure 2, Panel C).

Simple effects indicated nicotine-induced increases in both low and high sensation seekers, with a larger magnitude increase observed in low sensation seekers. In addition, high sensation seekers reported higher ratings of elation than low sensation seekers after smoking the 0.05-mg cigarette (p = .07). While group differences were found on ratings of vigor, with high sensation seekers reporting higher ratings than low sensation seekers, no main effects of nicotine yield or nicotine yield by group interactions were observed. Nicotine effects on other POMS scales did not differ as a function of sensation-seeking status. Figure 3 (Panel E) presents the pooled ratings of total positive. A main effect of nicotine yield was observed, with ratings at the 0.9-mg nicotine yield significantly increased compared with the 0.

05-mg yield. Similar effects were observed on ratings of friendliness. Nicotine yield-dependent decreases were observed on ratings Brefeldin_A of anxiety, anger, and depression. Addiction Research Center Inventory. The effects of nicotine yield on the PCAG Scale varied as a function of sensation-seeking status, with simple effects indicating a nicotine-yield dependent decrease among high sensation seekers only. In addition, high sensation seekers reported lower PCAG scores for both the 0.05- and 0.9-mg nicotine yields compared with low sensation seekers.

Both PGF and PGE2 produced by the CL increase the survivability of bovine luteal cells by suppressing FAS expression and CASP-mediated apoptosis [11]. In addition, since PGF stimulates P4 production in cultured bovine luteal cells selleck compound [10], luteal PGF is thought to be a luteoprotective factor [11]. On the other hand, both intramuscular PGF injection [37] and endogenous uterine PGF [38] induce luteolysis. Moreover, luteal PGF stimulated by exogenous or uterine PGF has been demonstrated to promote luteolysis in the cow [39]. Therefore, there is still some controversy about whether PGF has different roles in the luteal and uterine origins. In the present study, the viability of luteal cells treated with LH was much higher than that of untreated cells, while the increase in viability caused by LH was decreased by indomethacin (INDO, a COX inhibitor).

In addition, INDO decreased LH-increased PGF production, but did not affect P4 production in bovine luteal cells. The above findings suggest that the luteoprotective actions of LH are also mediated by PG production. Interestingly, although LH in combination with OP stimulated PGF production and LH in combination with INDO stimulated P4 production, both of these combinations decreased luteal cell viability in the present study. These results suggest that luteoprotective roles of LH require both P4 and PGF actions. Further studies are needed to clarify the exact mechanisms of luteoprotective actions of LH. In the present study, LH stimulated the expressions of COX-2 and PGFS but not the expressions of COX-1 and PGES.

In fact, LH stimulated PGF Drug_discovery production but not PGE2 production. Furthermore, LH stimulated the CBR1 enzyme that converts PGE2 to PGF. This could be the reason why the PGE2 concentration in the medium was not increased, although COX-2 expression was stimulated by LH. In addition, since PGF stimulates P4 secretion in bovine luteal cells [10, 11], LH may stimulate P4 secretion not only directly but also by stimulating PGF. However, INDO did not affect LH-increased P4 production in the present study. Since LH has been demonstrated to increase P4 production by a variety of signaling molecules other than PGF [6,7,8], the increased level of P4 production could be mainly induced by LH rather than by LH-stimulated PGF. In summary, LH stimulated P4 and PGF production but not PGE2 production. LH increased luteal cell viability, and this luteoprotective action of LH was inhibited by OP as well as by INDO. The overall findings suggest that LH protects CL function by stimulating the production of P4 and PGF in cows.

Those studies for which published data has been augmented by additional information are so labelled in Table 1. Studies used assays with widely varying cut-offs for the detection of HBV (Table 1). This could have introduced bias, with the use of more sensitive assays resulting in an apparent lower rate of suppression. However plotting the proportion www.selleckchem.com/products/kpt-330.html undetectable against the logarithm of the cut-off value showed no clear pattern (Figure 3) and the cut-off was ignored in further analyses. Figure 3 Log of HBV viral load assay cut-off against proportion undetectable at one year. The overall proportion suppressed was 57.4% (95% CI: 53.0�C61.7%), 79.0% (95% CI: 73.6�C83.8%), and 85.6% (95% CI: 79.2�C90.7%) after one, two, and three years of treatment with TDF (Table 3). Table 3 Suppression at yearly time points.

It was possible to assess rates of virological suppression by HBeAg status for patients from ten of the included studies. [6], [8], [13], [14], [19], [21]�C[23], [25], [27] For HBeAg positive and negative patients respectively the proportion fully suppressed was 51.8%, 82.0%, 86.6% and 76.3%, 82.1%, 75.0% at one, two and three years (Figure 4). After one year of treatment, a higher proportion of HBeAg negative than HBeAg positive individuals had a fully suppressed HBV viral load (p=0.005). However, after one year the rates of suppression were not significantly different. Figure 4 Percentage with undetectable HBV viral load over time, by HBeAg status. Table 4 shows the effects of prior and concomitant 3TC/FTC on virological suppression.

Effects are given for all patients and also stratified by prior or concomitant treatment with 3TC/FTC as appropriate. Overall, at one year prior exposure to 3TC had an odds ratio of 0.69 (95% CI: 0.45 to 1.08) and treatment with 3TC/FTC in addition to TDF of 1.24 (95% CI: 0.68 to 2.24), neither being statistically significant. The effect of prior exposure to 3TC/FTC was similar, but also not statistically significant, at each of one, two, and three years. The effect of concomitant treatment with 3TC/FTC favoured dual therapy at one year but TDF monotherapy at years two and three, but these effects were again not statistically significant. The odds ratios in the stratified analyses were similar to the effects overall but with even wider confidence intervals. There was no evidence of an interaction between prior and concomitant 3TC/FTC treatment (p=0.98 at 1 year, p=0.14 at 2 years and p=0.99 at 3 years). Between-study heterogeneity, Dacomitinib allowing for the effects of prior and concomitant 3TC/FTC treatment, was significant (p<0.01) at year 1 but not at year 2 (p=0.48) or at year 3 (p=1.0).