3), phosphorylation of Crh and HPr at Ser46 was strongly inhibited in the untreated cells (no additional glucose added) when buy DAPT growth ceased, i.e. after 9 h incubation (Fig. 4b, top panels). In contrast, much higher amounts of Crh~P and HPr(Ser)~P were detectable at that time (9 h) in the cells that were supplemented with additional glucose (Fig. 4b, compare lanes 3 and 10 in the top and bottom panels). This result unequivocally shows that exhaustion of the carbon source glucose prevents phosphorylation of Crh and HPr

by HPrK/P when cells enter the stationary growth phase. In this work, we analyzed the dynamics of phosphorylation of Crh in response to different nutritional conditions in vivo. Previous in vitro studies suggested that Crh becomes (de)-phosphorylated by HPrK/P at residue Ser46 like its homolog HPr, but whether this also applied to in vivo conditions was not clear. Our data confirm that

HPrK/P is actually the kinase responsible for phosphorylation of Crh in vivo (Fig. 2). Thus, one might expect a similar dynamics Selleckchem NVP-BEZ235 of phosphorylation of Crh and HPr at their Ser46-sites. Overall, this was indeed the case, but with some remarkable deviations. As expected, both Crh~P and HPr(Ser)~P levels decreased drastically or even disappeared when cells entered the stationary growth phase (Fig. 3). Exhaustion of the carbon source is responsible for accumulation of the non-phosphorylated proteins in this growth phase (Fig. 4). Consequently, stationary cells are released from CCR and primed for the uptake and utilization of alternative carbon sources. The degree to which Crh became phosphorylated during exponential growth depended on the quality of the carbon Acyl CoA dehydrogenase source. The various substrates could be classified into two

distinct groups, triggering the formation of either low or very high levels of Crh~P (Fig. 2). Such a splitting of the carbon sources into two distinct groups has not been observed previously in the formation of HPr(Ser)~P. In this case, a more gradual transition between the various substrates was detected (Singh et al., 2008). Nonetheless, the carbon sources that trigger either very low or very high levels of phosphorylation are the same for both proteins. Only a little Crh~P and HPr(Ser)~P is formed (Fig. 2; Singh et al., 2008) when cells utilize succinate, ribose or gluconate. Consequently, these gluconeogenic carbon sources cause no or only weak CCR (Singh et al., 2008). Except for gluconate, these substrates also yield slower growth rates in comparison with the other tested substrates (Fig. 2a; Singh et al., 2008). In contrast, high Crh~P as well as HPr(Ser~P) levels were detectable when a substrate of the PTS (glucose, fructose, mannitol, salicin, sucrose), sorbitol or glycerol was the carbon source (Fig. 2; Singh et al., 2008). Accordingly, all these sugars, which exert a strong CCR, enter the upper branch of the EMP pathway directly (Singh et al., 2008).

tuberculosis H37Rv. The best activity was found with one with a C10 chain length. This bactericidal activity can partly be attributed to its ability to damage the robust and complex cell envelope of Mycobacteria. Moreover, our study reveals the ability of decanol to attenuate biofilm formation by M. smegmatis. This knowledge can be used to develop new therapeutics and disinfectants

against mycobacteria. Tuberculosis is caused by Mycobacterium tuberculosis and results in innumerable deaths worldwide. Mycobacterium tuberculosis is highly infectious and is able to establish persistent infection in individuals even with a healthy immune STA-9090 order system and thus has infected more than one-third of the world’s population. The emergence of multidrug-resistant (MDR) and extremely drug-resistant tuberculosis strains (XDR) and its coinfection with HIV has made tuberculosis more difficult to treat (Global tuberculosis control, full report, 2011). The search for antimycobacterial agents for both therapy and disinfection is therefore now of major research interest across the world. In this regard, research Palbociclib supplier describing new antimycobcaterial agents from natural and synthetic sources is being reported with different target sites and mode of actions (Koyama et al., 2010; Kakwani

et al., 2011; Lougheed et al., 2011). Different types of alcohols, their derivatives and some lipophilic or amphiphilic compounds are known to exhibit antimycobacterial

activity (Júnior et al., 2009; Rugutt & Rugutt, 2011; Falkinham et al., 2012). Among the different class of alcohols, aliphatic alcohols are the most widespread compounds occurring naturally in plants and foods. There have been a number of reports on the antibacterial activities of long-chain fatty alcohols (Lansford et al., 1960; Sheu & Freese, 1973; Mates, 1974; Kato et al., 1978; Kato & Shibasaki, 1980; Kubo et al., 1993a, b; Tanaka et al., 2002; Kabelitz et al., 2003; Togashi et al., 2007). These studies showed that the antimicrobial activity is influenced Urease by the number of carbon atoms present in the alkanol chain and can be modulated by the presence of an effective head group that alters its polarity. The presence of double or triple bonds and their position can also play a crucial role in determining their antimicrobial activities (Ravel et al., 1955; Lansford et al., 1960; Sheu & Freese, 1973; Mates, 1974; Kabelitz et al., 2003). In addition, the type of organism and its cell-wall composition also influence the anti-microbial activity of a particular agent. However, no antimyobacterial activity of these long-chain fatty alcohols has previously been reported. In this context, in the present study we have investigated the effect of long-chain fatty alcohols against Mycobacterium smegmatis mc2155 and M. tuberculosis H37Rv.

, 1967). Ledoux et al. (1974)

expanded the experiments and found that the donor DNA from thiamine plus E. coli and not that from a thiamine minus mutant bacterial strain converted the plant to a thiamine plus condition. These results could not be reproduced in St. Louis or elsewhere (Lurquin, 2001), even with seeds and DNA provided by Ledoux. At one stage, Ledoux suggested that maybe cosmic gamma irradiation in the airplanes during trans-Atlantic flights inactivated the activity. Lurquin (2001) provides this website untestable alternative hypotheses that the data were faked by Ledoux himself or by a staff member in Belgium who wished to please his boss. This is the only one of the five cases we are considering in this report where cheating is thought (but not proven) to have occurred. For the other four, it was probably merely self-delusion, which is the definition of beyond the fringe science. A recent example indicating that beyond the fringe science is alive and still with us came with the claim that arsenic could ‘substitute for’ or ‘replace’ phosphorus in the DNA of a newly isolated bacterial strain (Wolfe-Simon et al., learn more 2011; published in ScienceExpress online on 2 December 2010). A blog posting criticizing the Wolfe-Simon

et al.’s article appeared 2 days later, on 4 December 2010 (http://rrresearch.fieldofscience.com/2010/12/arsenic-associated-bacteria-nasas.html). Then, science writer Carl Zimmer (http://www.slate.com/articles/health_and_science/science/2010/12/this_paper_should_not_have_been_published.html) headlined ‘This paper should not have been published’ and ‘Scientists see fatal flaws in NASA study of arsenic-based life’ just 5 days after online publication in Science. Zimmer used inverted commas to indicate that the first headline was a quote from a source (cited at the bottom of his sixth paragraph). The Slate article covers all of the basic reasoning showing that the article was beyond PLEKHB2 the fringe. There were other published criticisms. For example, Silver & Phung (2011) wrote a brief summary 3 weeks after

the initial online publication stating that the report was ‘science fiction’ rather than science. Much later, Matthew Herper in Forbes magazine headlined ‘New science papers prove NASA failed big time in promoting supposedly Earth-shaking discovery that wasn’t’ at http://www.forbes.com/sites/matthewherper/2012/07/08/new-science-papers-prove-nasa-failed-big-time-in-promoting-supposedly-earth-shaking-discovery-that-wasnt/. The scientific community usually does not use popular magazines as sources of understanding (although beyond the fringe science frequently does), and blogs and tweets are new. However, for arsenic in DNA, it was the authors and their government-funding agency NASA that used rapid Internet vehicles to communicate their ideas, which were then uniformly judged as beyond the fringe. What had happened? Wolfe-Simon et al.

, 1967). Ledoux et al. (1974)

expanded the experiments and found that the donor DNA from thiamine plus E. coli and not that from a thiamine minus mutant bacterial strain converted the plant to a thiamine plus condition. These results could not be reproduced in St. Louis or elsewhere (Lurquin, 2001), even with seeds and DNA provided by Ledoux. At one stage, Ledoux suggested that maybe cosmic gamma irradiation in the airplanes during trans-Atlantic flights inactivated the activity. Lurquin (2001) provides Talazoparib price untestable alternative hypotheses that the data were faked by Ledoux himself or by a staff member in Belgium who wished to please his boss. This is the only one of the five cases we are considering in this report where cheating is thought (but not proven) to have occurred. For the other four, it was probably merely self-delusion, which is the definition of beyond the fringe science. A recent example indicating that beyond the fringe science is alive and still with us came with the claim that arsenic could ‘substitute for’ or ‘replace’ phosphorus in the DNA of a newly isolated bacterial strain (Wolfe-Simon et al., buy Ixazomib 2011; published in ScienceExpress online on 2 December 2010). A blog posting criticizing the Wolfe-Simon

et al.’s article appeared 2 days later, on 4 December 2010 (http://rrresearch.fieldofscience.com/2010/12/arsenic-associated-bacteria-nasas.html). Then, science writer Carl Zimmer (http://www.slate.com/articles/health_and_science/science/2010/12/this_paper_should_not_have_been_published.html) headlined ‘This paper should not have been published’ and ‘Scientists see fatal flaws in NASA study of arsenic-based life’ just 5 days after online publication in Science. Zimmer used inverted commas to indicate that the first headline was a quote from a source (cited at the bottom of his sixth paragraph). The Slate article covers all of the basic reasoning showing that the article was beyond Rebamipide the fringe. There were other published criticisms. For example, Silver & Phung (2011) wrote a brief summary 3 weeks after

the initial online publication stating that the report was ‘science fiction’ rather than science. Much later, Matthew Herper in Forbes magazine headlined ‘New science papers prove NASA failed big time in promoting supposedly Earth-shaking discovery that wasn’t’ at http://www.forbes.com/sites/matthewherper/2012/07/08/new-science-papers-prove-nasa-failed-big-time-in-promoting-supposedly-earth-shaking-discovery-that-wasnt/. The scientific community usually does not use popular magazines as sources of understanding (although beyond the fringe science frequently does), and blogs and tweets are new. However, for arsenic in DNA, it was the authors and their government-funding agency NASA that used rapid Internet vehicles to communicate their ideas, which were then uniformly judged as beyond the fringe. What had happened? Wolfe-Simon et al.

, 2001; Wang et al., 2006), but until now the molecular differences between these species as well as their potential capacity of inbreeding are largely unknown. Therefore, tools for Tuber species’ discrimination are still needed to avoid frauds in the truffle market. The intraspecies gSSH experiment in O. maius yielded, after subtraction of O. maius OmMa3 with O. maius OmMa2 genomic DNA and reverse selleck screening library dot blot analysis, 16 specific sequences: five were single independent sequences, whereas 11 formed three contigs (Table 3; accession numbers HN262662–HN262669). Of the singletons, one showed similarity to an l-galactonate dehydratase,

one to a short-chain dehydrogenase/reductase family protein and three found no similarity in databases. Of the contigs, one showed similarity to glutathione synthetase, one to acetoacetyl-coenzyme A synthetase and one found no similarity. OmMa3 and OmMa2 are two isolates derived from a serpentine soil, characterized by a high content in chromium and nickel (Vallino et al., 2011). These two isolates are genetically distinct, on the basis of genetic fingerprinting, and show different abilities to grow in the presence of heavy metals, OmMa3 growing considerably better than OmMa2 on Ni- and Cr-amended media (Vallino et al., 2011). Heavy metal tolerance is a trait of particular interest for documenting genetic changes during adaptation, as heavy metal toxicity

represents a strong directional selective pressure resulting in the substitution of tolerance alleles at some loci (Willems et al., 2007). The genetic basis of heavy metal tolerance is not fully understood, and the questions on how KU-57788 purchase many genes are involved and on the dynamics of the alleles of these genes are still open. It is tempting to speculate that the sequences we have identified may represent genetic differences underlying different tolerance of the two isolates, but further investigations are needed. Interestingly, glutathione synthetase, the second enzyme in the glutathione

biosynthetic pathway, is known to be involved in metal tolerance (Pócsi et al., 2004; Reisinger et al., 2008). Glutathione plays a key role not only in metal detoxification but also in protecting cells from other environmental stresses, such as oxidative stress and xenobiotics (Memon & Schröder, 2009). Moreover, a recent study on Drosophila by Ortiz et al. (2009) Sclareol suggests that polymorphisms in GSH biosynthetic genes may be an important contributor to differential arsenic sensitivity. Therefore, this genomic region is a good candidate for further analyses on the genetic basis of metal tolerance in fungal isolates. In conclusion, our results show that gSSH is a quick and rather inexpensive approach that allows the identification of genomic differences both among (e.g. Tuber) and within (e.g. O. maius) fungal species. The sequences obtained by gSSH may be useful to identify species or strains as well as to investigate the genome plasticity, adaptation and evolution.

, 1980; Lang et al., 2004; Lee et al., 2007). These findings imply that the absence of an ipsilateral inhibitory response with weak TMS reflected the failure of CC neurons to Atezolizumab be excited, even though crossed CST neurons were excited. This notion is consistent with previous findings demonstrating that the threshold to induce TCI was higher than the RMT for contralateral MEPs (Ferbert et al., 1992; Trompetto et al., 2004). The stability of bimanual cyclic movement with different coordination conditions has been expressed by dynamic pattern theory, such as the Haken–Kelso–Bunz model (Haken et al., 1985; Schöner

& Kelso, 1988). Based on this model, the phase shift between left and right cycles critically affects the stability of bimanual action. However,

INK 128 nmr the bistable characteristic can be observed at low frequency; the bimanual action is stable at both in-phase and 180° out-of-phase. In the present study, the participants performed the symmetric and asymmetric force tracking tasks with almost equivalent accuracy, although synchrony of the left–right tracking trajectory was slightly lower during the asymmetric condition. This suggests that performance degradation due to bimanual constraint in the asymmetric force coordination was relatively low and was compensated for by the strategy of bimanual regulation, which was different from that in the symmetric condition. On the basis of this context, it may be that the observed modulation of TCI was due to an aspect of neural organization necessary for implementing a motor strategy to evade the constraints imposed on bimanual actions. As previous studies demonstrated, a lack of transcallosal communication leads to either deterioration (Serrien et al., 2001; Kennerley et al., 2002) or improvement (Franz et al., Montelukast Sodium 1996; Eliassen et al., 1999; Diedrichsen et al., 2003) in bimanual task performance according to the respective requirement for spatiotemporal coordination. That is, the functional importance of transcallosal neural communication depends on whether the coordination of left- and right-sided movements is required. In support

of this, we recently observed that TCI modulation was influenced by the coordination requirement of left and right hands during a bimanual task (T. Tazoe, S. Sasada & T. Komiyama, unpublished observation). Following this line, as our experiment was not designed to manipulate the required coordination between the symmetric and asymmetric conditions, two different interpretations may be possible for our findings of TCI modulation. One is that, during the asymmetric condition, the inhibitory effect between the motor cortices decreased, uncovering the excitatory interhemispheric neural communication. The CC is reported to have both excitatory and inhibitory transcallosal circuits (Asanuma & Okuda, 1962; Ugawa et al., 1993; Hanajima et al., 2001; Bäumer et al., 2006).

We also evaluated the extent to which researchers

attended to communication by examining whether publications included information on the time pharmacists spent delivering 3-MA chemical structure the intervention or the number of subsequent contacts. We found that nine studies[17–23,25,31,34–37,39] included information about the duration of pharmacist–patient interactions and 13 studies[17–19,21,22,26–39] recorded the number of follow-up visits between pharmacists and diabetic patients. Only six studies reported both the duration of pharmacist–patient interactions and the number of follow-ups.[17–20,22,31,34–37,39] To evaluate the extent of researchers’ attention to communication, we considered how pharmacists had been trained to deliver interventions. Only six studies reported that pharmacists had been trained in drug and disease management[22,26,27,29,30,32,34–38] while three stated that pharmacists

had been trained in patient-centred communication.[19,20,29,30,32] One study design[32] included, for example, ‘role-play’ exercises. In another case[29,30] pharmacists were involved in ‘experience-based learning’ SB431542 cell line to enable them to better understand diabetic patients’ experiences of shopping, exercising and blood-sugar self-testing. In three studies, authors reported that participating pharmacists had been provided with training in research protocol.[21,22,24] Finally,

one study reported that pharmacists had been taught the principles of patient-centred care through training in Self-Regulatory Model (SRM) theory.[19,20] Pharmacists in this project were specifically instructed, for example, to ‘give information, advice or reassurance in response to the patient’s expressed needs’ (p.166). The authors of this study Resminostat also audio-recorded a sample of the interventions for quality-control purposes. Quality control was defined as checking for ‘safety’ and as evaluating pharmacist’s advice as ‘helpful’ or not from the point of view of an expert review panel. Interventions were also documented as ‘useful’ or not from the point of view of patient participants. This study was also the only one that reported on having recorded actual communication between pharmacists and diabetic patients. The authors also reported that pharmacists had been specifically trained to listen to diabetic patients. Some researchers appear to presume that pharmacists practice patient-centred care as a result of their professional training as pharmacists. When researchers did not report that participating pharmacists had been specifically trained to deliver interventions according to patient-centred communication principles, researchers described pharmacists in three ways: as ‘diabetes educators’, ‘clinical or consultant pharmacists’ or simply as ‘pharmacists’.

Proportion of patients treated outside of clinical trials for non-genotype 1 who receive therapy with pegylated interferon and ribavirin Proportion of patients treated for non-genotype 1 with a Metavir score of F4 who are offered treatment with pegylated interferon and ribavirin unless contraindicated Proportion of patients with non-genotype 1–4 referred to a tertiary centre Proportion of patients not receiving therapy undergoing repeat non-invasive staging of their liver disease within 1 year The response rate of genotype 4 HCV monoinfection to a PEG-IF/RBV regimen is similar to that seen with genotype

1, with a figure ranging between 43–50% being observed in clinical trials. As with genotypes 2 and 3, neither of the two currently available HCV protease inhibitors has SB431542 research buy been studied, but the newer anti-HCV agents are being studied across all genotypes with excellent

initial responses in monoinfected patients [101]. Due to the low rates of success with pegylated Y-27632 molecular weight interferon and ribavirin we suggest that treatment is deferred where possible and treatment with newer agents within clinical trials actively sought. Where the individual has liver disease staging suggestive of Metavir stage 4, a complication of disease, or it is the informed wish of the patient to commence therapy, then treatment is recommended. This should be with pegylated interferon and ribavirin. The duration of therapy should be 48 weeks if an undetectable HCV RNA is achieved at 4 weeks, with a consideration to extend this to 72 weeks if achieved by 12 weeks. If the RNA is still detectable at 12 weeks, consideration should be given to discontinuing treatment. All individuals deferring therapy should undergo hepatic elastography or an alternative non-invasive test at least annually. Individuals infected with genotypes other than 1–4 should be referred to a centre with experience of treating HCV infection with these genotypes for a treatment

plan to be made in consultation with the host centre. We recommend patients without a decrease Oxymatrine of 2 log10 in HCV RNA at week 4 post diagnosis of acute infection (1D) or with a positive HCV RNA week 12 post diagnosis of acute infection (1C) are offered therapy. We recommend therapy be commenced prior to an estimated duration of infection of 24 weeks (1D). Patients who have not commenced treatment by this time should be managed as for chronic hepatitis C. We recommend all patients be offered combination therapy with pegylated interferon and weight-based ribavirin (1C). We recommend against treatment with PEG-IFN monotherapy (1C). We recommend treatment is discontinued if patients do not achieve an EVR (1C). We recommend patients with re-emergent virus after spontaneous or therapeutic clearance are assessed for relapse or reinfection (1C). We recommend patients with AHC who relapse are managed as for chronic hepatitis C (1D).

0; not significant). Among all participants, 35 (7%) reported genital ulcers and 34 (7%) reported genital discharge over the 6-month period. For the 70 participants with a recent STI diagnosis, only 19 (27%) indicated that this was their first non-HIV STI since testing HIV positive, 35 (50%) had had two previous STIs, and 16 (23%)

stated that this was their third or fourth STI since testing HIV positive. Among participants with an STI diagnosis, 16 (24%) reported genital ulcers at the time of the assessment and 20 (34%) were having genital discharge. The most frequently diagnosed STIs were herpes simplex virus (HSV) infection (n=26; 37%) and syphilis (n=25; 36%). In addition, nine (13%) participants Alectinib research buy reported having been diagnosed with Z-VAD-FMK order gonorrhoea, 14 (20%) had chlamydia, and four (6%) were diagnosed with nonspecific urethral infection. Comparisons of the demographic and health characteristics of participants who had not been diagnosed with a recent STI and those who had been diagnosed are shown in Table 1. Three out of four participants were receiving antiretroviral therapy, and treatment was proportional among those who had not and who had been diagnosed with a recent STI. Participants who had had a recent STI were significantly younger and had fewer years of education than their counterparts who had not been diagnosed with an STI. Individuals with a recent

STI had experienced more HIV-related symptoms, had lower CD4 cell counts, and were significantly more likely to be unaware of their viral load and less likely to indicate having an undetectable viral load. Individuals who were recently diagnosed with an STI also demonstrated significantly greater alcohol use, including higher rates of problem drinking on the AUDIT. Nonalcohol drug use was far less common in the sample. However, participants who had a recent

STI were more likely to have used cannabis in the previous 3 months (Table 2). Analyses examining sexual behaviours with all partners showed that participants recently diagnosed with an STI had significantly more partners, more Sucrase protected intercourse, and more total intercourse than participants who had not been diagnosed with a recent STI. There were no effects of participant viral load and there were no STI × viral load interactions for sexual behaviours across all partners (Table 3). Results for sexual behaviours with non-HIV-positive partners demonstrated a different pattern. There was a main effect for viral load on protected sexual acts and on total sexual acts; participants with a detectable viral load reported significantly greater rates of protected and total sexual acts. There was also a main effect for having contracted an STI on number of non-HIV-positive partners; participants who contracted an STI reported a greater number of partners.

For MI events, the IRR (95% CI) compared with never smokers decreased from 3.73 (2.46, 5.64) within the first year of having stopped smoking to 3.00 (1.84, 4.88) at 1–2 years, 2.62 (1.42, 4.83) at 2–3 years, and 2.07 (1.19, 3.63) at >3 years. Similarly, the IRR for CHD events decreased from 2.93 (2.07, 4.14) in the first year of having stopped smoking to 2.48 (1.65, DAPT molecular weight 3.73) at 1–2 years, 1.90 (1.09, 3.29) at 2–3 years and1.83 (1.16, 2.89) at >3 years. The IRR (95% CI) also decreased for CVD

events from 2.32 (1.69, 3.18) within the first year of having stopped smoking to 1.84 (1.25, 2.70) at 1–2 years, 1.60 (0.99, 2.61) at 2–3 years and 1.49 (0.99, 2.24) at >3 years (Table 2 and Figure 1). Compared with current smokers, the risk of MI, CHD and CVD among patients who stopped smoking for >3 years was reduced by approximately 30% [IRR (95% CI) 0.61 (0.36, 1.04) for MI, 0.74 (0.48, 1.15) for CVD, and 0.68 (0.46, 1.01) for CHD] (Table 2). There were 1902 deaths reported during follow-up, yielding a crude rate of 12.54 (95% CI

11.98–13.11) Fulvestrant purchase per 1000 person-years. Table 3 provides crude death rates per 1000 person-years for specific smoking status groups and IRRs for previous, current and stopped smoking groups compared with the never smoked group. Unlike those for the CVD events, these IRRs did not decrease linearly with increased time since smoking cessation. In a post hoc mortality analysis in which we aimed to demonstrate a clearer mortality signal in a subgroup at higher risk of mortality, we restricted the analysis to patients aged >50 years during follow-up. In this group, a total 634 deaths were recorded (crude rate of 19.64 per 1000 person-years). Again, there was no decreasing trend IRR for each additional year of having stopped smoking (Table 3 and Fig. 1). The risks of death overall and for those aged >50 years were similar for patients

who stopped smoking for >3 years Glutamate dehydrogenase compared with current smokers (Table 3). One explanation for the lack of a reduction in mortality following smoking cessation is that patients stopped smoking following diagnosis of a serious illness. To investigate this hypothesis further, we summarized causes of death by smoking status. Overall, HIV/AIDS was recorded as the underlying cause in 27% of deaths, CVD in 10%, chronic viral hepatitis in 13%, non-AIDS-related malignancies in 12%, invasive bacterial infection in 6%, and other in 24%. A larger proportion of never smokers died from HIV/AIDS (35%) compared with previous smokers (27%), current smokers (23%) and those who stopped smoking (29%). Of those who died, a greater proportion of previous smokers and those who had stopped smoking during D:A:D follow-up had non-AIDS-related malignancies as the reported underlying cause of death (17% for both groups) compared with the never smokers and current smokers (10% for both groups).