Bacillus cereus ATCC 14579 cold-adaptation genes were identified by a transposon mutagenesis strategy. One of the cold-sensitive mutants harbours a transpositional insertion upstream of the BC3118 encoding a putative P450-cytochrome enzyme. This small protein of 86 aa is conserved in all the available genomes of the B. cereus group, but no specific function was assigned to this enzyme and no GSK 3 inhibitor link was described with low-temperature adaptation. A second set of mutants presented defects in growth at 10 °C: in those

two mutants, transposon interrupted the two genes BC3773 and BC3774, encoding two subunits of the PFOR. This enzyme catalyses the coenzyme A (CoA)-dependent oxidative decarboxylation of pyruvate with a ferredoxin as the electron acceptor (Charon et al., 1999). The role of the PFOR in B. cereus cold adaptation is not clear. Bacteria in general modulate membrane fluidity by altering their fatty acid composition (Mansilla et al., 2004). Absence of the PFOR activity may lead to a decrease in the quantities of acetyl-CoA, one of the precursor metabolites of the fatty acid synthesis. By the reverse reaction, mutation of PFOR activity could also decrease the amount of pyruvate produced from acetyl-CoA, leading to a decrease in the biosynthesis of aa such as alanine, valine PR-171 purchase and leucine, with an impact

on protein synthesis. All these mutants showed impaired growth at a low temperature, but were also affected by salt or acid stresses. In contrast, Pregnenolone the 9H2 mutant showed a marked and interesting cold-sensitive phenotype through a delayed growth at 10 °C compared with WT, but was neither impaired in its growth at optimal temperature nor under the various stressful conditions tested. The 9H2 cell morphology at 30 °C was similar to the WT. However, at 10 °C, the 9H2 mutant formed long filamentous cells, while WT did not, suggesting that BC0259 could contribute to regular cell division of B. cereus at a low temperature,

as proposed for E. coliΔcsdA cells (Jones et al., 1996). The BC0259 product was annotated in the B. cereus ATCC 14579 genome as an RNA helicase with nine motifs that constitute the conserved helicase core and a DEAD box, revealing that it belongs to the DEAD-box subgroup of the helicase superfamily 2 (Bleichert & Baserga, 2007). RNA helicases participate in many aspects of RNA metabolism (Silverman et al., 2003) and some have been shown to be cold-induced (Jones et al., 1996; Chamot et al., 1999; Lim et al., 2000).The 9H2 mutant harbours a transpositional insertion upstream of the BC0259 gene, which reduced BC0259 gene expression during adaptation at 10 °C, but not at 30 °C, as revealed by qRT-PCR experiments.

Bacillus cereus ATCC 14579 cold-adaptation genes were identified by a transposon mutagenesis strategy. One of the cold-sensitive mutants harbours a transpositional insertion upstream of the BC3118 encoding a putative P450-cytochrome enzyme. This small protein of 86 aa is conserved in all the available genomes of the B. cereus group, but no specific function was assigned to this enzyme and no selleck kinase inhibitor link was described with low-temperature adaptation. A second set of mutants presented defects in growth at 10 °C: in those

two mutants, transposon interrupted the two genes BC3773 and BC3774, encoding two subunits of the PFOR. This enzyme catalyses the coenzyme A (CoA)-dependent oxidative decarboxylation of pyruvate with a ferredoxin as the electron acceptor (Charon et al., 1999). The role of the PFOR in B. cereus cold adaptation is not clear. Bacteria in general modulate membrane fluidity by altering their fatty acid composition (Mansilla et al., 2004). Absence of the PFOR activity may lead to a decrease in the quantities of acetyl-CoA, one of the precursor metabolites of the fatty acid synthesis. By the reverse reaction, mutation of PFOR activity could also decrease the amount of pyruvate produced from acetyl-CoA, leading to a decrease in the biosynthesis of aa such as alanine, valine PD-0332991 molecular weight and leucine, with an impact

on protein synthesis. All these mutants showed impaired growth at a low temperature, but were also affected by salt or acid stresses. In contrast, PJ34 HCl the 9H2 mutant showed a marked and interesting cold-sensitive phenotype through a delayed growth at 10 °C compared with WT, but was neither impaired in its growth at optimal temperature nor under the various stressful conditions tested. The 9H2 cell morphology at 30 °C was similar to the WT. However, at 10 °C, the 9H2 mutant formed long filamentous cells, while WT did not, suggesting that BC0259 could contribute to regular cell division of B. cereus at a low temperature,

as proposed for E. coliΔcsdA cells (Jones et al., 1996). The BC0259 product was annotated in the B. cereus ATCC 14579 genome as an RNA helicase with nine motifs that constitute the conserved helicase core and a DEAD box, revealing that it belongs to the DEAD-box subgroup of the helicase superfamily 2 (Bleichert & Baserga, 2007). RNA helicases participate in many aspects of RNA metabolism (Silverman et al., 2003) and some have been shown to be cold-induced (Jones et al., 1996; Chamot et al., 1999; Lim et al., 2000).The 9H2 mutant harbours a transpositional insertion upstream of the BC0259 gene, which reduced BC0259 gene expression during adaptation at 10 °C, but not at 30 °C, as revealed by qRT-PCR experiments.

Fig. S1. Erlotinib cost Multiple sequence alignments generated by

clustalw analysis of the N-termini of MtrB homologs identified in the genomes of 22 metal-reducing Shewanella strains. Fig. S2. Multiple sequence alignments generated by clustalw analysis of the N-termini of three CXXC-containing MtrB paralogs identified in the Shewanella oneidensis genome. Fig. S3. Growth of Shewanella oneidensisMR-1 wild-type (●), ∆mtrB (∆), C42A (□), and C45A (×) mutant strains with either O2 (A), DMSO (B), TMAO (C), fumarate (D), nitrite (E), thiosulfate (F), or nitrate (G) as electron acceptor. Table S1. Amino acid sequence identity (ID), similarity (Sim), expect-value (e-value), N-terminal CXXC motif (CXXC motif), number of amino acid residues

in the N-terminus (N-term length), and number of β-sheets in the C-terminus (No. β-sheets) of the MtrB homologs identified in the genomes of 22 metal-reducing Shewanella strains. Table S2. Amino acid sequence identity (ID), similarity (Sim), expect-value (e-value),108mm N-terminal CXXC motif (CXXC motif), number of amino acid residues in the N-terminus (N-term length), and number of β-sheets in the C-terminus (No. β-sheets) of the three MtrB paralogs identified in the genome of Shewanella oneidensis MR-1. Table S3. Phylogenetic affiliation (Class), amino acid sequence identity (ID, %), similarity (Sim, %), expect-value (e-value), N-terminal CXXC motif, (CXXC motif) number of amino selleck chemical acid residues in the N-terminus (N-term length), number of β-sheets in the C-terminus (No. β-sheets), and reported dissimilatory metal reduction or oxidation activity of the host strain (metal redox) for 52 MtrB homologs displaying similarity to Shewanella oneidensisMtrB. “
“Carboxy (C)-terminal processing proteases (CTP) are a relatively new group of serine proteases. Found in a broad range of organisms – bacteria, archaea, algae, plants and animals – these

proteases are involved in the C-terminal processing of proteins. In comparison with amino-terminal processing of bacterial proteins, less is known about C-terminal processing and its physiological function. Bacterial CTPs appear to 2-hydroxyphytanoyl-CoA lyase influence different basal cellular processes. Although CTPs of Gram-negative bacteria are generally referred to as being localized in the periplasm, there is little experimental evidence for this. We show for the first time the subcellular localization of a CTP-3 family protein from Pseudomonas aeruginosa, named CtpA, in the periplasm by a carefully designed fractionation study. Our results provide experimental evidence for the generally accepted hypothesis that CTPs are located in the periplasmic space of Gram-negative bacteria. Carboxy (C)-terminal processing proteases (CTP) form a relatively new group of serine proteases that are involved in the C-terminal processing of proteins.

The ROC curve analysis showed the accuracy of EBV load in PBMC1 for predicting progression to B lymphoma: the area under the ROC curve was

0.72 (95% CI 0.58–085; selleck chemical P = 0.0014). The optimal cut-off threshold of EBV load that yielded the maximal sensitivity and specificity for predicting the development of B lymphoma was determined as 3.2 log10 copies/106 PBMCs. The sensitivity and specificity obtained with this cut-off value were 75 and 65%, respectively. Setting the cut-off level at 2.8 log10 copies/106 PBMCs allowed a higher sensitivity (85%) to be obtained, but at the price of a substantial decrease in specificity (40.5%). Having an EBV DNA load > 3.2 log10 copies/106 PBMCs was associated with an adjusted OR of 4.82 (95% CI 1.33; 17.46) for progression to B lymphoma within the next 10 months. EBV load in PBMCs had, as expected, poor accuracy

for identifying patients at risk for developing brain lymphoma (area under ROC curve 0.57; 95% CI 0.38; 0.76; P = 0.5). In this study, high levels of EBV DNA in PBMCs were predictive of subsequent progression to systemic B lymphoma when measured within 1 year before the diagnosis of lymphoma. One previous study failed to demonstrate such Selleck GSK3 inhibitor an association between EBV DNA level and the development of ARL [16]. This could be explained by the fact that, in this previous study, patients and controls were not matched for CD4 cell count and also by the fact that a limited number of cases (n = 9) and controls (n = 12) were tested. As expected, we found no association between EBV DNA levels in PBMCs and/or

in serum and the development of cerebral lymphoma. The lack of sensitivity of EBV PCR in blood Gemcitabine for the diagnosis of cerebral lymphoma in patients with AIDS was demonstrated by Fan et al. [19], although there was a correlation between PBL and high levels of EBV DNA in cerebrospinal fluid [20]. This is likely to be attributable to the fact that PBLs are very compartmentalized tumours with low or no systemic dissemination. This result, however, has no major implications in clinical practice, as the incidence of PBL has dramatically fallen following the introduction of cART, PBL now representing less than 10% of all ARLs [7]. Indeed, no case of PBL was reported after 1995 in our cohorts. In this study, EBV DNA was quantified in stored samples (i.e. mainly cryopreserved PBMCs and serum samples). The choice of biological material used for EBV DNA quantification, i.e. PBMCs, whole blood, plasma or serum, has been a matter for debate and there is currently no consensus on which is the best material for EBV load measurement in EBV-associated lymphoproliferative disorders [21, 22]. In these settings, whole blood or PBMCs more frequently contained amplifiable EBV DNA than did plasma or serum [15, 23-25].

The ROC curve analysis showed the accuracy of EBV load in PBMC1 for predicting progression to B lymphoma: the area under the ROC curve was

0.72 (95% CI 0.58–085; buy BGB324 P = 0.0014). The optimal cut-off threshold of EBV load that yielded the maximal sensitivity and specificity for predicting the development of B lymphoma was determined as 3.2 log10 copies/106 PBMCs. The sensitivity and specificity obtained with this cut-off value were 75 and 65%, respectively. Setting the cut-off level at 2.8 log10 copies/106 PBMCs allowed a higher sensitivity (85%) to be obtained, but at the price of a substantial decrease in specificity (40.5%). Having an EBV DNA load > 3.2 log10 copies/106 PBMCs was associated with an adjusted OR of 4.82 (95% CI 1.33; 17.46) for progression to B lymphoma within the next 10 months. EBV load in PBMCs had, as expected, poor accuracy

for identifying patients at risk for developing brain lymphoma (area under ROC curve 0.57; 95% CI 0.38; 0.76; P = 0.5). In this study, high levels of EBV DNA in PBMCs were predictive of subsequent progression to systemic B lymphoma when measured within 1 year before the diagnosis of lymphoma. One previous study failed to demonstrate such selleck chemical an association between EBV DNA level and the development of ARL [16]. This could be explained by the fact that, in this previous study, patients and controls were not matched for CD4 cell count and also by the fact that a limited number of cases (n = 9) and controls (n = 12) were tested. As expected, we found no association between EBV DNA levels in PBMCs and/or

in serum and the development of cerebral lymphoma. The lack of sensitivity of EBV PCR in blood STK38 for the diagnosis of cerebral lymphoma in patients with AIDS was demonstrated by Fan et al. [19], although there was a correlation between PBL and high levels of EBV DNA in cerebrospinal fluid [20]. This is likely to be attributable to the fact that PBLs are very compartmentalized tumours with low or no systemic dissemination. This result, however, has no major implications in clinical practice, as the incidence of PBL has dramatically fallen following the introduction of cART, PBL now representing less than 10% of all ARLs [7]. Indeed, no case of PBL was reported after 1995 in our cohorts. In this study, EBV DNA was quantified in stored samples (i.e. mainly cryopreserved PBMCs and serum samples). The choice of biological material used for EBV DNA quantification, i.e. PBMCs, whole blood, plasma or serum, has been a matter for debate and there is currently no consensus on which is the best material for EBV load measurement in EBV-associated lymphoproliferative disorders [21, 22]. In these settings, whole blood or PBMCs more frequently contained amplifiable EBV DNA than did plasma or serum [15, 23-25].

This protein was then purified to learn more homogeneity by affinity chromatography using an immobilized Zn2+ matrix. The purified fusion protein was subjected to specific cleavage by thrombin. The resulting tag-free SarA (14.7 kDa) was analyzed by gel electrophoresis and used

in all further experiments. Previous work in the laboratory (Debarbouille et al., 2009) showed that analysis of the genome sequence of S. aureus revealed the presence of two Ser/Thr kinases in S. aureus, which were overproduced and purified. The phosphorylating activity of SarA was first assayed by incubating the pure protein in the presence of radioactive ATP. After gel migration and autoradiography, no labeled band was detected, indicating that His6-SarA was unable to autophosphorylate (Fig. 2a and b, lane 2). In contrast, when Stk1 or SA0077 was added to the incubation medium, His6-SarA was intensely labeled (Fig. 2a and b, lanes

4 and 7). Furthermore, to ensure that the 16-kDa band was actually SarA, the histidine tag was previously removed from SarA, and the tag-free protein was then incubated with both the kinases. Once more, we did observe a shifted band (14 kDa) corresponding to this protein (Fig. 2a and b, lanes 5 and 8), indicating that the native SarA was effectively phosphorylated. The phosphorylation of SarA by Stk1 or SA0077 was then studied in more detail by analyzing its phosphoamino acid content. Under the conditions used, only acid-resistant phosphoamino acids were analyzed because a number of other phosphorylated compounds, such as phosphohistidine, phosphoarginine and phosphoaspartate, are known to be labile in acid. Interestingly, Galunisertib mw two-dimensional analysis of an acid hydrolysate of SarA showed that this protein was mainly phosphorylated at threonine residues by Stk1 [Fig. 2c(1)], whereas it was phosphorylated essentially at serine residues by SA0077 [Fig. 2c(2)]. SarA was previously described to bind several promoter regions to regulate different

genes involved in the virulence of S. aureus including accessory regulator gene promoter (P1sar) (Cheung et al., 2008a), accessory gene regulator promoter Sclareol (P2agr) (Chien & Cheung, 1998), regulator gene promoter (Prot) (Manna & Ray, 2007; Hsieh et al., 2008) and staphylococcal fibronectin gene promoter (PfnbA) (Wolz et al., 2000). To further investigate whether phosphorylated SarA interacts differently from nonphosphorylated SarA with its promoters, comparative gel shift assays were performed on promoters P2agr, PfnbA, Prot and P1sarA, with either unphosphorylated SarA or SarA phosphorylated by Stk1 (Fig. 3) or by SA0077 (Fig. 4). Concerning P2agr, a striking difference emerged between unphosphorylated SarA, which must be added in the amount of 8 μg to obtain a complete shift (Fig. 3a), and SarA phosphorylated by Stk1, where only 2 μg was sufficient to obtain the same shift (Fig. 3b).

2). Interestingly, the UV survival curve of the infectious B. burgdorferi 297 (clone BbAH130) wild-type strain used in the present study was likely similar to that of the infectious B. burgdorferi B31 clone (5A18NP1) used by Lin et al. (2009), but was distinctly different from that reported for the infectious B. burgdorferi B31M1 strain studied by Liveris et al. (2004, 2008). The reason for this difference is at present unclear, but may be strain-related, because the Cyclopamine in vivo design of our experiments and those of Liveris and colleagues was otherwise identical. In vitro growth of B. burgdorferi uvrA inactivation mutants was

inhibited by ROS but not by RNS. Dissociation of in vitro susceptibility to ROS and RNS has been reported to occur in a M. tuberculosis uvrB mutant (Darwin & Nathan, 2005). In this case, the mutant was Y-27632 order more susceptible to RNS than the wild-type parent

but showed similar susceptibility to ROS. It was not possible to examine the in vivo function of B. burgdorferi uvrABbu because ΔuvrABbu and its derivatives, in contrast to the parental strain, lacked lp25 (Purser & Norris, 2000; Iyer et al., 2003) (data not shown) and were noninfectious. Studies are currently underway to develop an infectious uvrA inactivation mutant in order to examine its in vivo virulence. Several lines of evidence suggest that the ability of B. burgdorferi to overcome DNA damage following phagocytosis is critical to its ability to survive and produce disease in the host. Mutants of mutS and mutS-II, genes whose products are involved in DNA mismatch repair, display decreased infectivity in Celastrol immunocompetent mice (Lin et al., 2009). Furthermore, resistance of B. burgdorferi to rapid killing in vitro by phagocytes has been correlated with in vivo infectivity (Georgilis et al., 1991). Although the majority of phagocytosed borrelia are rapidly killed after ingestion, some remain viable

and cultivable (Montgomery et al., 1993), and can stimulate a phagocytic oxidative burst (Georgilis et al., 1991). Plausibly, these few viable organisms could be sufficient to initiate infection of the mammalian host. In summary, homologous recombination and extrachromosomal complementation have been used to show that uvrABbu is needed to repair DNA damage in B. burgdorferi exposed in vitro to UV, ROS and MMC but not in B. burgdorferi exposed to RNS or low pH. M.S. and L.B.I. contributed equally to this work, which was supported by grant R01 AI 048856 to F.C.C. We would like to thank Dr M. Norgard, University of Texas Southwestern Medical Center, Dallas, TX, for providing B. burgdorferi 297 and clone BbAH130. We also thank Dr Julia Bugrysheva for advice. Figure S1. Confirmation of DNA structure and expression of mRNA in Borrelia burgdorferi 297 and derivatives. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors.

5% clinical and parasitologic response at 28 days.13 However, a larger recent trial in Papua New Guinea

of learn more 195 children with vivax malaria treated with different artemisinin-based combination therapies compared with conventional chloroquine-sulfadoxine-pyrimethamine (CQ-SP) demonstrated adequate clinical and parasitologic response of only 69% in the dihydroartemisinin-piperaquine group compared to 13% in the CQ-SP group.14 In summary, CRPV is emerging as a clinically significant issue among travelers with imported malaria. Awareness of epidemiology and a detailed travel exposure are critically important to the recognition of CRPV. Mefloquine is an effective treatment for patients potentially infected with CRPV, and treatment strategies for P. vivax may eventually need to be reconsidered if CRPV becomes more widespread. Further research is needed to elucidate the mechanisms of resistance and to validate better prospective assays for chloroquine resistance. Malaria prophylaxis for travel to destinations with CRPV may not require change if P. falciparum is the predominant clinical concern, but an expanded role for primaquine in prevention could be considered.15 Pre-travel advice to travelers going Alpelisib molecular weight to such destinations should include discussion of CRPV and the risk of resistance and/or relapse. The authors state they have no conflicts of

interest to declare. “
“Splinter hemorrhages appear in a variety of conditions. One identified cause is ascent to altitude, but trauma and extreme conditions have Resveratrol been thought to be responsible. We document the appearance of splinter hemorrhages in a group of adults during several days of easy touring at an altitude of 11,000 feet (3,350 m). Splinter hemorrhages are seen in conditions of varying severity

including (but not limited to) infective endocarditis, vasculitis, the antiphospholipid syndrome, chronic meningococcemia, ingestion of tyrosine kinase inhibitors, trauma,[1] and activities of daily living (especially in the elderly).[1, 2] Chronic[4] or acute[5, 6] exposure to high altitude has also been associated with this finding, but, in this scenario, extreme conditions and trauma have been thought to play a causative role. This report describes splinter hemorrhages associated solely with ascent to moderately high altitude and in the absence of associated trauma or extreme conditions. This 71-year-old physician presented for evaluation of numerous splinter hemorrhages (Figure 1). He denied fever, chills, muscle or joint pains, chest pain, difficulty breathing, or neurologic symptoms. He had no known heart murmur, and was in general good health, with hypertension, well controlled on hydrochlorthiazide and atenolol, and diabetes, well controlled on metformin 500 mg daily (hemoglobin A1c = 5.6). He had just returned from a 7-day trip to Peru where he spent 2 days in Cuzco (altitude 11,000 feet) and 1 day in Machu Picchu (altitude 8,000 feet).

The concentration of DNA in negative controls was measured at 260 nm using

a NanoDrop spectrophotometer. The PCR mixture (25 μL) SCH727965 was composed of 12.5 μL of 2 × Combi-PPP mix (Top-Bio Ltd, Prague, Czech Republic, contains hot start-Taq DNA polymerase, 5 mM MgCl2, buffer, deoxyribonucleotides and loader), 0.5 μL 10 μM forward primer, 0.5 μL 10 μM reverse primer, 0.5 μL DNA template and 11 μL water. Thermal programs for primer pairs used in this study are given in Table 1. Nested PCR directed to ITS region was performed using primer pair NSI1/NLB4 in the first amplification and either the pair Tu1sekvF/Tu2sekvR or the pair UncI/UncII in the second. Nested PCR directed to the β-tubulin gene was performed using primers Bt2a/Bt2b in Linsitinib datasheet the first amplification and primers tubtubf/elytubr in the second. The annealing temperature originally recommended for

this primer pair is 63 °C, but with this temperature the PCR was not sufficiently sensitive for T. aestivum DNA and the annealing temperature was therefore decreased as indicated in Table 1. In addition, nested PCR was performed using the primers Bt2a/BTAEMB-R in the first amplification and BTAE-F/Bt2b in the second. The same thermal program as indicated for the primer pair BTAE-F/BTAEMB-R in Table 1 was used in both steps of amplification but the annealing temperature was set to 56 °C. The product of the first amplification was always diluted 1 : 100 before being used as a template in the second amplification. Templates were prepared by the Carnitine palmitoyltransferase II addition of small amounts of T. aestivum DNA (extracted from the sample S13, see Appendix S1) into complex nontarget DNA (negative control A). Resulting mixtures contained 2.5, 0.25, 0.025, 0.0025,

0.00025 or 0.000025 ng S13 DNA and 24.5 ng nontarget DNA in 1 μL water. These mixtures were used in nested PCR with primer pairs NSI1/NLB4 (first amplification) and Tu1sekvF/Tu2sekvR (second amplification) as indicated above with annealing at 59 °C. A 5-μL aliquot of the product of PCR amplified using the Tu1sekvF/Tu2sekvR primer pair was mixed with 9 μL water, 1 μL buffer R and 5 U of TaiI restriction endonuclease (New England Biolabs Inc., Ipswich, MA). The mixture was then incubated for 3 h at 65 °C and immediately separated on agarose gel. Soil and ectomycorrhizae samples were collected in the native habitat of T. aestivum, Chuchelský háj, near Velká Chuchle, Prague, Czech Republic. Plant cover was dominated by Carpinus betulus with addition of Fraxinus excelsior, Corylus avellana and Tilia cordata seedlings. Twelve 200 g soil samples were collected on an L-shaped terrain transect at 1 m equidistant points (Fig. 1) from the depth of 0–10 cm (A-horizon, rendzina on silurian lime). Ectomycorrhizae were separated manually from the soil sample.

“
“Glutamatergic inputs to the nucleus accumbens (NAc) modulate both appetitive and fearful motivation. It has been suggested that pathological disturbances of glutamate signaling in NAc contribute to motivation disorders, ranging from excessive desire in drug addiction to paranoia in schizophrenia. Metabotropic glutamate receptors are of special interest, as metabotropic Group II receptor (mglu2/3) agonists have been proposed as potential treatments for both addiction and schizophrenia. Here we tested whether local mglu2/3

receptor blockade Selleckchem GDC-0980 in the medial shell of the rat NAc can generate intense distortions of motivation or affect, which might model clinical selleck kinase inhibitor dysfunction. We found that microinjection of the mglu2/3 antagonist LY341495 at sites throughout medial shell suppressed appetitive motivation to eat and drink. Simultaneously, LY341495 microinjections generated fearful motivation in the form of defensive

treading or burying. To assess whether the valence shift extended into a parallel hedonic shift from affective ‘liking’ to ‘disliking’ we employed the taste reactivity test, which measures affective orofacial reactions to the sensory pleasure or displeasure of tastes. We found that LY341495 microinjections reduced positive ‘liking’ reactions to sucrose and enhanced ‘disliking’ reactions. Overall, mglu2/3 antagonism at most shell sites produced a similar valence shift from positive to negative. This pattern comprised (i) generation of fearful behaviors, and (ii) induction of aversive affective reactions, together with (iii) loss of appetitive ingestion and (iv) loss of ‘liking’ for rewards. These results are discussed in terms of implications

for clinical disorders and the influence of corticolimbic glutamate PAK6 inputs to NAc in the generation of motivation and affect. “
“Maternal rhythms entrain the prenatal and neonatal circadian clock in the suprachiasmatic nucleus (SCN) before light entrainment is established. However, the responsible time cues for maternal entrainment are not identified. To examine the role of cyclic changes of ambient temperature in maternal entrainment, blind neonatal rats carrying a clock gene (Per2) bioluminescence reporter were exposed to either of three ambient temperatures (10, 20 or 30 °C) during 6-h maternal separation in the early light phase. Cold exposure was performed from postnatal day 1 (P1) to P5. On P6, the SCN was harvested and cultured for photometric monitoring of the circadian rhythm in Per2 expression. Here we demonstrate that the daily cold exposure phase-delayed the circadian Per2 expression rhythms at P6 in a temperature-dependent manner. Exposure to 10 °C produced the largest phase-shift of 12.7 h, and exposure to 20 and 30 °C yielded moderate shifts of 4.1 and 4.5 h, respectively.