While, specific parental behaviours such as Parents’ perceived ability to

withhold frequent cariogenic snacks from their children even when they fussed for Small molecule library supplier it was inversely associated with the presence of dental decay in their child. Not all beneficial practices, however, had beneficial effects on dental caries; in this study, the frequency of tooth-brushing and/or tooth-brushing with supervision did not have a positive influence on the child’s caries experience. Although this agrees with some studies[27, 28], others have reported lower caries levels associated with frequent tooth-brushing[20, 29]. The controversial results and conclusions may be due to acidogenicity of biofilm or poor tooth-brushing techniques of children and/or their caregivers.

Interestingly, none of the factors mentioned in this Sotrastaurin section were significantly associated with dt/ds, implying the role of other more important indicators when assessing caries severity. Nevertheless, the information derived from both Gao et al.’s (2010)[4] and this study provides practical guidelines to steer health promotion efforts to specifically target certain knowledge and practices, especially for children and parents with higher caries rate in Singapore. Because of the perceived discomfort of many individuals with the disclosure of their family income, the type of dwelling was chosen to measure the socio-economic status (SES) in this study. In this study, the caries experience was not consistently associated with the type of dwelling, a relationship that has been otherwise well documented in other published reports[4, 30]. The inconsistent association could have been a function of the sampling from the public health medical clinics, which itself may be selective for patients from the lower socio-economic group. The utilization Sclareol of the type of housing may also be a crude measure for the measurement

of socio-economic status in Singapore as it does not account for the extremely high housing cost in Singapore (e.g., more than 50% of the population live in government housing developments) as well as other social and cultural factors that may be unique in this country (e.g., extended family units etc). The limitations of this study include intra-operator reliability, small sample size, convenience sampling, the potential underestimation of caries experience because only a visual-tactile examination, without radiographs, was employed, and the innate inaccuracies in the answers encountered in the interviewer-administered questionnaire (e.g., truthful answers). Improvements to the current questionnaire could be made in future studies by the inclusion of specific questions with regard to fluoride intake (e.g.

In experimental viral infection, cholesterol also falls before TG rises [12]. TG levels were higher in HIV-positive patients at

an earlier stage of HIV disease. A decrease in the levels of cholesterol, in particular HDLC, also occurred in HIV-positive patients long before hypertriglyceridaemia occurred. In HIV-positive patients, cholesterol levels fell before TG levels rose. LDLC levels were significantly lower in HIV-positive patients compared with controls only when CD4 lymphocyte counts were<50 cells/μL. TG levels were higher and TC levels lower, compared with controls, in HIV-positive patients with low CD4 lymphocyte counts (<50 cells/μL and 50–199 cells/μL) and in those with active OIs. The atherogenicity index (TC:HDLC and LDLC:HDLC ratios) was significantly higher in HIV-positive patients than in control subjects. The authors thank all subjects who gave their informed consent to participate in this study. "
“The aim of the study ICG-001 solubility dmso was to determine the aetiology and clinical predictors of peripheral

lymphadenopathy in HIV-infected individuals during the antiretroviral (ARV) era in a nontuberculosis endemic setting. A multicentred, retrospective cohort study of peripheral lymph node biopsies in HIV-positive AUY-922 mouse adults was carried out. A total of 107 charts were identified and reviewed for clinical features, lymphadenopathy size, and ARV use and duration. Biopsy results were categorized, and multivariate logistic regression determined independent predictors of lymphadenopathy aetiology. Evaluation of 107 peripheral lymph node biopsies revealed that 42.9% of peripheral lymphadenopathy was attributable to malignancy, 49.5% to reactive changes, and 7.5% to infections,

with only 2.8% of all cases secondary to tuberculosis. Fevers, weight loss, ARV use, and lower viral loads are significantly associated with nonreactive lymphadenopathy. Lymphadenopathy is likely to be reactive or malignant in nontuberculosis endemic regions. Readily available clinical features can aid clinicians in predicting the underlying aetiology, those at risk for malignancy, and who to biopsy. “
“We studied the influence of noninjecting and injecting drug use on mortality, dropout rate, and the course of antiretroviral www.selleck.co.jp/products/AG-014699.html therapy (ART), in the Swiss HIV Cohort Study (SHCS). Cohort participants, registered prior to April 2007 and with at least one drug use questionnaire completed until May 2013, were categorized according to their self-reported drug use behaviour. The probabilities of death and dropout were separately analysed using multivariable competing risks proportional hazards regression models with mutual correction for the other endpoint. Furthermore, we describe the influence of drug use on the course of ART. A total of 6529 participants (including 31% women) were followed during 31 215 person-years; 5.1% participants died; 10.5% were lost to follow-up.

8) and vocal sounds as deviants (P = 0.2), while the reverse was true for the other two blocks. In each block, standards always consisted of just one token of one sound category (for example, just a female voice), while deviants consisted of both tokens of the opposite sound category (for example, a cello and a French Horn). Both tokens of the deviant sounds occurred Lumacaftor research buy equiprobably (P = 0.1 each). Both standards and deviants were of two durations – 350 and 550 ms, with each duration occurring equiprobably (P = 0.5). The 200-ms difference between short and long sounds used in the current study is similar to that used in previous studies employing the auditory distraction paradigm (e.g. Schröger

& Wolff, 2000; Mager et al., 2005). NVP-LDE225 research buy Each block consisted of 200 trials (160 standards and 40 deviants). The inter-stimulus interval varied randomly between 1.6 and 2 s. The ROT condition was identical to the NAT condition, with the only difference being that spectrally-rotated versions of voices and musical sounds were used throughout. Table 1 lists all conditions and blocks of the study. Figure 2 details the structure of a single block. The eight blocks of the experiment were presented in a Latin square design, lasting approximately 6 min each. Participants were instructed to press one

response button for short sounds and another for long sounds. Three examples of short and three of long sounds present in each block were always played at the beginning of a block to familiarize participants with the sounds they were instructed to categorize. Hand to response button mapping was counterbalanced across participants. Importantly, unlike in odd-ball paradigms, responses were provided for all sounds (i.e. standards and deviants). The N1 ERP component elicited by the onset of auditory stimuli was evaluated in order to compare the two groups’ early neural processing of musical and vocal sounds. N1 is a measure of early sensory encoding of the physical properties of sound, such as frequency, complexity and intensity

(Näätänen & Picton, 1987). Importantly, previous ERP research has demonstrated the influence of musical training on the amplitude of N1 and its N1c subcomponent. For example, it has been shown to be enhanced in musicians O-methylated flavonoid in response to both musical notes and pure tones, with greater enhancement for the sounds of the instrument of training (e.g. Pantev et al., 1998, 2001; Shahin et al., 2003; Baumann et al., 2008). We therefore predicted that musicians would exhibit a larger N1 component to musical sounds. We also speculated that given the similarity of vocal and musical timbres and their underlying acoustic properties, musicians might also show an enhanced N1 to voices. Additionally, we have evaluated several behavioral and electrophysiological measures associated with distraction.

We cannot draw conclusions in this regard based on our results because of the elevated percentage of samples in which IL-6 plasma levels were under the limit of detection, as has been seen in other studies [10, 15]. Lipid disturbances have also been investigated in relation to the increased cardiovascular risk in patients undergoing cART interruption, although the results are somewhat contradictory. KU57788 Chronic infection, including that produced by HIV, is associated with changes in lipoprotein metabolism. This can lead to proatherogenic dyslipidaemia, especially hypertriglyceridaemia, and decreased HDL-c and LDL-c,

associated with changes in the properties of lipids, rendering them more proatherogenic [23]. In accordance with previous cART interruption studies, we found a decrease in total-c and LDL-c, but also in HDL-c [4-6]. As a result, see more in our study no change in the total-c/HDL-c ratio in patients discontinuing cART was found, in contrast to the SMART study in which an unfavourable change was observed [4]. As far as we know, this is the first study in which patients were treated mainly with NNRTIs, and our data are, at least in part, consistent with

those of the SMART study, in which the strongest HDL-c reduction was found in patients receiving NNRTIs [4]. As has been described, we observed a strong negative correlation between viral load and lipid measurements, supporting a role for HIV in these variables [6]. An interesting finding of our study, described previously in a non-HIV-infected [24] and HIV-infected population [25], is the association between lipid parameters,

especially HDL-c, and MCP-1 and sVCAM, confirmed in the multivariate Tyrosine-protein kinase BLK analysis and maintained over the lengthy follow-up period. Experiments with inflammatory lipopolysaccharide-induced animal models have shown that treatment with ApoA-I, the major component of HDL-c, induces a decrease in MCP-1 and sICAM-1. ApoA-I has modulating effects on MCP-1 expression [26]. Furthermore, it is known that the antioxidant effect occurring through paraoxonase-1, an enzyme contained in HDL-c, inhibits MCP-1 synthesis by endothelial cells [27]. It is likely that the anti-inflammatory effects of HDL-c are attenuated in untreated HIV infection. The negative correlation found between HDL-c and endothelial biomarkers is consistent with the results of studies pointing to a close association between lipids and inflammation pathways, probably mediated by HIV itself [25]. Our study has some limitations, the most important being the small sample, although significant differences were found between arms in some of the parameters. Baseline CD4 cell count differed between arms; however, the role of CD4 count in determining biomarker concentrations has not been clearly documented in previous interruption studies [5-10].

, 2001), and comX (∆SMcomX) (Li et al., 2002) were used in this study. The comS (∆SMcomS) and the comR (∆SMcomR) mutants were constructed using a nonpolar ligation

PCR mutagenesis method described previously (Lau et al., 2002). Streptococcus mutans strains were grown at 37 °C with 5% CO2 in either Todd-Hewitt broth (Becton Dickinson, MD) containing 0.3% yeast extract (Difco Laboratories) (THYE), or chemically defined medium (CDM) described previously (Mashburn-Warren et al., 2010). Erythromycin Regorafenib price and spectinomycin were used as needed at concentrations of 10 μg mL−1 and 1 mg mL−1, respectively. sXIP and synthetic CSP (sCSP) peptides were synthesized using F-MOC chemistry (Advanced Protein Technology Centre, Hospital for Sick Kids, Toronto, ON, Canada). Stock concentrations of 1 μM of sXIP and 0.4 mM sCSP were prepared

in DMSO and water, respectively. Growth kinetics were monitored using an automated growth reader (Bioscreen C; Labsystems, Finland) as previously described (Senadheera et al., 2007). Overnight cells grown in THYE were pelleted, washed, and resuspended in phosphate buffered saline (1× PBS). The resuspended culture was diluted 1 : 50 using prewarmed THYE or CDM and grown to an OD600 ~ 0.1. Next, 1 μg mL−1 of the donor plasmid DNA (pDL277; specR) (LeBlanc et al., 1992) was added to 1-mL aliquots of the culture in the presence or absence of CSP (0.4 μM) or XIP (10 μM), and samples were incubated for 90 min. For XIP, control cultures containing 1% DMSO GW-572016 molecular weight were utilized. After incubation, cultures were serially diluted and plated on THYE plates with and without antibiotics. TF was calculated as transformant colony-forming units (CFUs) divided by the total number of viable CFUs, times 100. Overnight cultures in THYE were pelleted, washed, resuspended in sterile 1× PBS, and diluted 1 : 50 using warm THYE or CDM. Each suspension was supplemented with either 2 μM CSP or 10 μM XIP. Cultures without peptides Megestrol Acetate or containing 1% DMSO

were used as controls. All cultures were grown to an OD600 ~ 0.8, at which point, the cells were gently sonicated on ice and used for viability assays. Cells were serially diluted, plated on THYE agar, and CFUs counted. Results standardized using cellular dry weight. These standardized values were then used to calculate the percentage survival by dividing the standardized number of viable cells after treatment by the standardized total number of cells without peptide, times 100. Overnight cultures of UA159 in THYE were pelleted, washed, resuspended in sterile 1× PBS, and diluted 1 : 20 using warm CDM. The subcultures were allowed to grow to an OD600 of 0.4, after which they were split into two where one was exposed to 1% DMSO and the other to 10 μM XIP. Cultures were further incubated, and samples were taken at varying time points (0, 1, 2, 3, 4, and 5 h) after exposure to XIP, gently sonicated, serially diluted, and plated on THYE plates for CFU determination.

Nevertheless, deeper into the gingival connective tissue, gingipain concentrations become gradually lower and stimulate, rather than inhibit, inflammation. This may in

turn induce connective tissue and bone destruction, which are the hallmarks of periodontitis. It is evident that P. gingivalis has developed mechanisms to invade and persist into the host, by astutely Panobinostat purchase adapting to its local niche. Its paradoxically opposing (stimulatory vs. inhibiting) effects on innate immune and inflammatory responses aim to subvert host defence mechanisms, in order to facilitate its survival in the tissues (Hajishengallis, 2009; Hajishengallis & Lambris, 2011). The net effect of this deregulated equilibrium is likely to determine if site-specific disease progresses beyond or remains at stationary phase. Whether inflammation is beneficial for P. gingivalis may depend on the stage of its establishment in the host (Hajishengallis, 2009; Pathirana et al., 2010). At early stages, suppression of inflammation and evasion of host recognition would aid P. gingivalis in colonizing, invading and establishing at the targeted site. At later stages, once P. gingivalis is well established, inducing inflammation may facilitate its increased demands in nutrients. Alternatively, P. gingivalis may

induce a ‘nonproductive inflammation’, one that fails to eliminate it, yet is sufficient to induce mediators of tissue destruction (Hajishengallis, 2009). Finally, as periodontitis is of polymicrobial

nature, it is reasonable to consider the role of different bacterial species within the context of (subgingival) Janus kinase (JAK) biofilm GPCR & G Protein inhibitor communities. Hence, P. gingivalis is likely to function in concerted action with other species, to their mutual benefit. For instance, complement manipulation by P. gingivalis may denote a coevolution strategy to support other species present in the biofilm, which may reciprocally provide further colonization opportunities and nutrient availability to P. gingivalis. Subsequent changes in the local microenvironment can differentially regulate expression of its virulence factors, and hence the proinflammatory or anti-inflammatory potentials of P. gingivalis. This is strongly indicated by recent evidence demonstrating that even at low abundance, this species qualitatively and quantitatively affects the composition of the oral commensal microbiota, which are in turn required for P. gingivalis-induced inflammatory bone loss (Hajishengallis et al., 2011). For these reasons, P. gingivalis is now considered a ‘keystone’ species in subgingival biofilms (Honda, 2011). This work was supported by the Institute of Oral Biology, Center of Dental Medicine, University of Zürich. “
“d-Xylulokinase catalyzes the phosphorylation of d-xylulose in the final step of the pentose catabolic pathway to form d-xylulose-5-phosphate.

2 °C, which confirms that strain MY14T does not belong to the genospecies O. flavum. The DNA–DNA relatedness studies between strains ND5 and H. saxobsidens NS11T, the strain with the highest 16S rRNA (99.8%) and cpn60 (98%) gene sequence similarity with ND5, showed that ΔTm values were 6.6 °C. However, the ΔTm value between H. glaciei UMB49T and strain ND5, sharing 99.6% 16S rRNA and 97.6%cpn60 gene sequence similarity, was 1 °C, which is well below the 5 °C cut-off point recommended for the delineation of species (Wayne Poziotinib price et al., 1987; Rosselló-Mora & Amann, 2001). Similar observations of contrasting high 16S rRNA gene sequence similarity (99.6–99.8%) and low (3–57%) DNA–DNA relatedness have

been reported among all described Herminiimonas-type strains (Kämpfer et al., 2006; Muller et al., 2006; Lang et al., 2007; Loveland-Curtze et al., 2009). On the basis of the results described above, it can be concluded that the strain ND5 (=NBRC 102664, =CCM 7665) is another strain of H. glaciei and strain MY14T represents a novel species within the genus Oxalicibacterium, for which the name O. solurbis sp. nov. is proposed. Oxalicibacterium solurbis (sol.ur’bis. L. n. solum, soil; L. n. urbs urbis, a city; N.L. gen. n. solurbis, of city soil, where the type strain was isolated). Gram-negative, small rods 0.4–0.5 × 0.8–1.2 μm (mean cell volume 0.07 μm3), motile by

polar flagella. At low temperatures around 4 °C, elongated cells are sometimes observed. No spores were observed. Oxidase FDA approved Drug Library datasheet and catalase reactions are positive. Colonies on nutrient agar (Oxoid CM3) with lactate are slightly yellow pigmented. Forms smooth, glistening,

raised, opaque with entire edges; the diameter is up to 0.5–1 mm after 3 days of incubation at 28–30 °C. Growth occurs at 4 °C and up to 37 °C, but not 42 °C. Optimum growth occurs at 37 °C and pH 8.0. Grows in media containing 5% NaCl. The specific growth rate (μ) under optimum conditions with lactate was 0.14 h−1. No acid produced from glucose. Nitrate is not reduced to nitrite. Negative for indole production, arginine dihydrolase, urease, esculin, casein Anacetrapib and gelatine hydrolysis and β-galactosidase. Sugars and alcohols were not utilized; utilizes fumarate, glycolate, dl-lactate, l-malate, malonate (weak), pyruvate, succinate, oxalate, l-alanine and l-glycine. Other differential characteristics are given in Table 1. The main fatty acids are C16:0, C17:0 cyclo, C19:0 cyclo ω8c and C10:0 3-OH. The major quinone system is ubiquinone Q-8. The major phospholipids are phosphatidylethanolamine and phosphatidylglycerol. The G+C content of DNA is 63.3 mol%. The type strain, MY14T (=NBRC 102665T, =CCM 7664T), was isolated from a soil sample using the membrane-filter enrichment technique. We are grateful to Dr Jean P. Euzéby, for his help with the Latin nomenclature of the species epithets. Thanks are also due to Dr J. Loveland-Curtze for supplying the type strain of H.

37) (Fig. 3c). To ensure that the decreased adhesion and invasion rate was a consequence of the fact that the Lcl antibodies covered Lcl and was not due to possible side effects of the antibodies, experiments were repeated with XlnC antibodies buy INK 128 of the same isotype as a control. The results obtained with the latter antibodies showed no difference in the adhesion and invasion of host cells compared with nontreated WT cells (Fig. 3d). To further exclude that masking

other adhesion factors caused by steric hindrance of bound antibodies might be the basis of the abovementioned results, Lcl-adhesion assays were performed with immobilized recombinant Lcl protein. Adhesion of the A549, macrophage-like cells and A. castellanii to the immobilized Lcl protein was influenced by preincubation of the protein film with Lcl-specific antibodies. The use of different antibody concentrations GW-572016 molecular weight demonstrated that the adhesion was specifically hindered by Lcl-specific antibodies, in an antibody concentration-dependent manner. The A549 cells showed an adhesion of 21% (P<0.001), 80% and 95% using 20, 2 and 0.2 μg Lcl-specific antibodies, respectively (Fig. 4a). The influence on the macrophage-like cell line was less pronounced, with a decrease of only

25% (P=0.06) using 20 μg of Lcl-specific antibodies (Fig. 4b). In contrast, no effect of antibody treatment was seen for the adhesion of A. castellanii to the immobilized film of Lcl, as similar results were obtained for the negative control (coated BSA) (Fig. 4c). In conclusion, the results of these incubation assays with Lcl-specific antibodies suggest that Lcl plays a role in the adhesion process of L. pneumophila. Coimmunoprecipitation experiments were performed to investigate the presence of possible partners on the host cells that interact with Lcl. The eukaryotic C1qR was suggested

to be a possible interaction partner, because it is involved in the phagocytosis pentoxifylline of microorganisms. This receptor interacts, for example, with the complement factor C1q and lung surfactant A through binding of the collagen-like region of these proteins, resulting in phagocytosis (Hoppe & Reid, 1994; Grubor et al., 2006). Moreover, the C1qR is present on both cell lines that were shown to interact with Lcl. Coimmunoprecipitation experiments using anti-C1qR antibodies and Lcl antibodies indicated an interaction between the Lcl protein of L. pneumophila Philadelphia and the C1qR of the A549 and the U937 cell line (Fig. 5). The previously described adhesion–infection assays were repeated with lung epithelial cells A549 and macrophage cell line U937 using the IPTG-inducible WT/pMMBNlcl, with 19 repeat units, and the WT/pMMBNlcl(14) strain, with 14 repeat units. The WT/pMMBNlcl(14) strain adhered to and invaded the lung epithelial cells significantly better (P=0.02) than WT/pMMBNlcl after 60 min.

3c). The β-glucosidase buy Fostamatinib activity was then measured under standard conditions in the presence of various metal ions (Zn2+, Mg2+, Co2+, Ca2+, and Mn2+). The activity appeared to be strongly inhibited in the presence of 5 mM Zn2+ or Co2+, which caused, respectively, a 64% or 70% activity drop. No ion tested had any positive effect

on the activity of the BglB protein. Kinetic experiments were performed by mixing the enzyme with different concentrations (0.25–10 mM) of pNPG. The Vmax and Km were determined by linear least squares fitting of a Lineweaver–Burke plot of the Michaelis–Menten equation. The BglB protein showed a Vmax of 5.8 μmol L−1 min and a Km of 1.34 mM. The substrate AZD6244 mw specificity of the BglB protein was tested on different substrates diluted in 100 mM sodium phosphate buffer pH 6.0 and incubated at 40 °C for 30 min (Table 3). The enzyme was found to hydrolyze both p-nitrophenyl-β-d-glucopyranoside and p-nitrophenyl-β-d-xylopyranoside, showing a strong preference for the xylopyranoside substrate, its specific activity being almost five times as high with this substrate than with pNPG. The enzyme failed to hydrolyze p-nitrophenyl-α-d-glucopyranoside, o-nitrophenyl-β-d-galactopyranoside, or p-nitrophenyl-β-d-cellobioside. β-Glucosidases are a major group among glycoside

hydrolases, belonging to EC 3.2.1.21 (Henrissat & Davies, 1997). They constitute a heterogeneous group of enzymes with various functions, and are found in many organisms (Esen, 1993): bacteria, fungi, plants, nonvertebrates, and vertebrates. Although they typically act upon β1–4 bonds linking two glucose or substituted glucose molecules, they can also remove other monosaccharides and are classified according to their substrate specificities. Some can thus perform the hydrolysis of cellobiose or cello-oligosaccharides Obatoclax Mesylate (GX15-070) to glucose, participating in the final step of cellulose depolymerization and thus ensuring its complete digestion. In insects, three substrate-specificity-based classes are distinguished. β-Glucosidase activity is

notably present in the digestive systems of many insects (Terra et al., 1996). In the guts of lower termites, it is produced largely by the salivary glands and the gut flora (Inoue et al., 1997). Focusing on the termite Neotermes koshunensis, Tokuda et al. (2002) detected 75% of the β-glucosidase activity in the salivary glands and 15% in the hindgut containing numerous flagellates. In particular, they found a β-glucosidase of glycoside hydrolase family 1 in the salivary glands (Tokuda et al., 2002). In termite guts, other microorganisms such as bacteria of genus Spirochaeta also show β-glucosidase activity (Dröge et al., 2006). Interestingly, the characterized β-glucosidase also displays β-xylosidase activity.

Both signals were expected to be represented in the amygdala

as this brain region is known to play a crucial role in mediating vigilance and attention (Davis & Whalen, 2001), and in learning to predict aversive outcomes (Schiller et al., 2008; Eippert et al., 2012). Additionally, we tested for unsigned PE effects in the midbrain as recent buy Idelalisib animal studies suggest that the surprise-induced enhancement of learning depends on the integrity of midbrain–amygdala connections (Lee et al., 2006, 2008, 2010). Twenty-two healthy male subjects (mean age 26.9 years, range 21–33 years) participated in this study. Due to equipment malfunction one subject had to be excluded from further analysis. Only male volunteers were enrolled in the study to reduce variability in amygdala activation based on gender effects in conditioning and other emotional tasks (Milad et al., 2006; Cahill, 2010). The study was approved by the Ethics Committee of the Medical Board in Hamburg. All participants gave written informed consent

Copanlisib concentration and were paid for their participation. The paradigm used was a classical Pavlovian delay-conditioning procedure including an acquisition and a reversal phase. Reversal of cue–outcome associations provided a characteristic test to assess associability as the outcomes became surprising again with the beginning of the reversal phase (Holland & Gallagher, 1999; Li et al., 2011). Abstract fractal images served as visual CSs and the US was an electric shock (150 ms duration, 20 pulses/s of 0.01–100 mA) delivered to the dorsum of the right hand. Before the experiment, the intensity of the US was individually adjusted to be aversive. For this purpose, volunteers received shocks of varying intensity. They rated the aversiveness of the shocks on a rating scale ranging from 1 (not aversive) to 10 (very aversive and not tolerable). The intensity of the shock that received the rating score

9 (very aversive, but still bearable) was selected as the final intensity administered throughout the experiment. We used three different visual cues, each presented 40 times throughout the entire experiment. During Aprepitant acquisition, cue A co-terminated with a shock in 20/20 occasions (100% reinforcement, CS100), cue B was paired with shock in 10/20 occasions (50% reinforcement, CS50) and cue C was never followed by the US (CS–). Thus, the acquisition phase comprised the first 20 occurrences of each cue. In the reversal stage, the CS100 was not paired with shock (new CS–), whereas the CS50 was followed by a shock in every trial (new CS100) and the CS– was followed by a shock on 50% of the occasions (new CS50). The reversal phase immediately followed the acquisition phase. It started after a fixed number of trials such that the 61st trial was always the first trial of the reversal phase. Again, each cue was presented 20 times leading to a total experimental time of approximately 40 min.