The supernatant was removed, and radioactivity in cells and supernatant was counted by liquid scintillation spectrometry. Internal pH was calculated from the distribution of 14C and 3H between the pellet and the supernatant. The accumulation of benzoic acid in E. ruminantium was abolished ABT-888 molecular weight by pretreatment of the

cells with 10 μM tetrachlorosalicylanilide, a protonophore (Hamilton, 1968), suggesting there was little or no active uptake or binding of benzoic acid by the cells. The chemical potential gradient (ZΔpH) generated by the pH gradient across the cell membrane was calculated from the Nernst relationship: Z = 2.3 RT/F or 62 mV at 39 °C. Intracellular volume was calculated using a separate aliquot of culture. One millilitre of culture was incubated with 3H2O (7 μCi, 125 μCi mL−1 and hydroxy [14C] methyl inulin (0.7 μCi, 11.1 mCi mmol−1) for 10 min, before centrifuging as before. The distribution of 14C-inulin and 3H2O in the pellet and the supernatant

allowed the exclusion volume of inulin compared to H2O to be calculated and hence the intracellular volume (Rottenberg, 1979). The electrical potential (Δψ) was calculated from the uptake of the lipophilic cation [phenyl-14C]tetraphenylphosphonium bromide (TPP+). One millilitre of culture was incubated under CO2 with TPP+ IDH inhibitor (0.05 μCi, 31 mCi mmol−1) and 3H3O (0.5 μCi, 16 μCi mL−1), then centrifuged and counted as before. Δψ was calculated from the distribution of TPP+ between the intra- and extracellular space (Rottenberg, 1979). The total transmembrane potential (Δp) was calculated as Δp = Δψ − ZΔpH. Nonspecific uptake/binding

of TPP+ was corrected by subtracting the apparent uptake in cells that had been treated with toluene (1% v/v, 1 h). Intracellular K+, Na+ and Ca2+ concentrations were measured in cells that had been centrifuged and resuspended in 25% TCA, then diluted in deionized water. Hydroxyl [14C] methyl inulin (0.7 μCi mL−1, 11.1 mCi mmol−1) ASK1 was added to the cultures before centrifugation to allow corrections to be made for extracellular medium trapped in the cell pellet. Na+ and K+ were analysed by atomic emission spectrometry on a Pye Unicam SP9 atomic absorbance spectrometer, while Ca2+ was determined by atomic absorbance on the same instrument. ATP pools were measured by a luciferase method (Wallace & West, 1982) in cells 2 h after the addition of the ionophores. Protein was determined using Folin reagent (Herbert et al., 1971). Tetronasin was a gift from Coopers Animal Health Limited, Berkhamsted, Herts. Monensin was from Sigma. TCS and TPP were gifts from I.R. Booth, University of Aberdeen. [Carboxy-14C]-benzoate was from New England Nuclear, Stevenage, Herefordshire. All other radiochemicals were from Amersham. The potency of monensin and tetronasin against E. ruminantium, S. bovis, P. albensis and L. casei was determined by inoculating bacteria into media in which the concentration of ionophore was serially doubled.

The supernatant was removed, and radioactivity in cells and supernatant was counted by liquid scintillation spectrometry. Internal pH was calculated from the distribution of 14C and 3H between the pellet and the supernatant. The accumulation of benzoic acid in E. ruminantium was abolished find more by pretreatment of the

cells with 10 μM tetrachlorosalicylanilide, a protonophore (Hamilton, 1968), suggesting there was little or no active uptake or binding of benzoic acid by the cells. The chemical potential gradient (ZΔpH) generated by the pH gradient across the cell membrane was calculated from the Nernst relationship: Z = 2.3 RT/F or 62 mV at 39 °C. Intracellular volume was calculated using a separate aliquot of culture. One millilitre of culture was incubated with 3H2O (7 μCi, 125 μCi mL−1 and hydroxy [14C] methyl inulin (0.7 μCi, 11.1 mCi mmol−1) for 10 min, before centrifuging as before. The distribution of 14C-inulin and 3H2O in the pellet and the supernatant

allowed the exclusion volume of inulin compared to H2O to be calculated and hence the intracellular volume (Rottenberg, 1979). The electrical potential (Δψ) was calculated from the uptake of the lipophilic cation [phenyl-14C]tetraphenylphosphonium bromide (TPP+). One millilitre of culture was incubated under CO2 with TPP+ CYC202 (0.05 μCi, 31 mCi mmol−1) and 3H3O (0.5 μCi, 16 μCi mL−1), then centrifuged and counted as before. Δψ was calculated from the distribution of TPP+ between the intra- and extracellular space (Rottenberg, 1979). The total transmembrane potential (Δp) was calculated as Δp = Δψ − ZΔpH. Nonspecific uptake/binding

of TPP+ was corrected by subtracting the apparent uptake in cells that had been treated with toluene (1% v/v, 1 h). Intracellular K+, Na+ and Ca2+ concentrations were measured in cells that had been centrifuged and resuspended in 25% TCA, then diluted in deionized water. Hydroxyl [14C] methyl inulin (0.7 μCi mL−1, 11.1 mCi mmol−1) second was added to the cultures before centrifugation to allow corrections to be made for extracellular medium trapped in the cell pellet. Na+ and K+ were analysed by atomic emission spectrometry on a Pye Unicam SP9 atomic absorbance spectrometer, while Ca2+ was determined by atomic absorbance on the same instrument. ATP pools were measured by a luciferase method (Wallace & West, 1982) in cells 2 h after the addition of the ionophores. Protein was determined using Folin reagent (Herbert et al., 1971). Tetronasin was a gift from Coopers Animal Health Limited, Berkhamsted, Herts. Monensin was from Sigma. TCS and TPP were gifts from I.R. Booth, University of Aberdeen. [Carboxy-14C]-benzoate was from New England Nuclear, Stevenage, Herefordshire. All other radiochemicals were from Amersham. The potency of monensin and tetronasin against E. ruminantium, S. bovis, P. albensis and L. casei was determined by inoculating bacteria into media in which the concentration of ionophore was serially doubled.

Implementation of pooling of RNA for acute HIV screening presents several challenges. The need to provide rapid turnaround of test results

in a clinically meaningful time frame to ensure patient follow-up makes it difficult to accumulate a large number of specimens for pooling [15]; this barrier may be overcome by pooling specimens from dried blood spots [24]. Optimal pool size depends on the prevalence of acute HIV infection in the population and the skill of the laboratory personnel if a manual pooling technique is required. The failure of rapid HIV tests in this study to identify all cases of chronic HIV infection led to an increased number of positive pools requiring additional testing that highlighted chronic rather than acute HIV cases. Patients with a negative or discordant rapid HIV test had ∼2% probability of having chronic HIV infection in this setting. From this study, we are unable to evaluate CYC202 concentration whether this relatively high false negative rate, higher than reported by the test kit manufacturers, was the result of operator error, faulty test kits/storage, or characteristics of the patient population. There was no apparent change during the study period in the rate of false negative results, despite retraining the HIV counsellors and changing the test kits. A recently reported

South African field study also noted challenges in HIV rapid test sensitivity compared with enzyme-linked immunosorbent assay and pooled HIV RNA PCR. In that Ipilimumab manufacturer study, which also used the SD Bioline kit, 5% of participants, all of whom were pregnant, had false negative results [25]. A high rate of false negative rapid tests was also reported in a study from South Africa among children on antiretroviral therapy, however, the test kits evaluated were different from those used in NADPH-cytochrome-c2 reductase the current study [26]. The performance

of rapid test kits has been disappointing in other contexts [22,27,28], suggesting that inaccurate rapid tests may not be a setting- or test-specific problem. Other than the Abbott Determine HIV 1/2 rapid test, none of the rapid kits used during the study period has been extensively validated against gold standard tests in Africa in published studies; the World Health Organization recommends that individual countries evaluate each assay used to determine its performance characteristics and suitability for use within a given setting [29,30]. To the extent that this is not practised, many false negatives are probably occurring, as the settings using pooled HIV RNA are extremely limited. Rapid HIV testing has been an essential element in improving diagnostic capacity and treatment opportunities for patients in resource-limited settings [31]. It is important to counsel patients and providers, however, that there is a small but real risk of a false negative test due to both chronic and acute infection and to encourage retesting; country-wide guidelines should recommend a retesting frequency to guide counsellors’ efforts.

The high-K+-evoked overflow of β-NAD+ is attenuated by cleavage of SNAP-25 with botulinum neurotoxin A, by inhibition of N-type voltage-dependent Ca2+ channels with ω-conotoxin GVIA, and by inhibition of the proton gradient of synaptic vesicles with bafilomycin A1, suggesting that β-NAD+ is likely released Venetoclax research buy via vesicle exocytosis. Western analysis demonstrates that CD38, a multifunctional protein that metabolizes β-NAD+, is present on synaptosomal membranes

and in the cytosol. Intact synaptosomes degrade β-NAD+. 1,N 6-etheno-NAD, a fluorescent analog of β-NAD+, is taken by synaptosomes and this uptake is attenuated by authentic β-NAD+, but not by the connexin 43 inhibitor Gap 27. In cortical neurons local applications of β-NAD+ cause rapid Ca2+ transients, likely due to influx of

extracellular Ca2+. Therefore, rat brain synaptosomes can actively release, degrade and uptake β-NAD+, and β-NAD+ can stimulate postsynaptic neurons, all criteria needed for a substance to be considered a candidate neurotransmitter in the brain. “
“In recent years, magnetic resonance imaging has allowed researchers to individuate the earlier morphological development of the right hemisphere compared with the left hemisphere during late-gestational development. Anatomical asymmetry, however, does not necessarily mean functional asymmetry, and Rucaparib order whether the anatomical differences Alectinib between hemispheres at this early age are paralleled by functional specialisations remains unknown. In this study, the presence of lateralised electrical brain activity related to both pitch detection and discrimination was investigated in 34 prematurely-born infants [24–34 gestational weeks (GWs)] all tested at the same post-conceptional age of 35 weeks. By means of a frequency–change oddball experimental paradigm, with ‘standard’ tones at 1000 Hz (P = 90%) and ‘deviant’ tones at 2000 Hz (P = 10%), we were able to record higher right event-related

potential activity in the interval windows between 350 and 650 ms after stimulus onset. An explorative hierarchical cluster analysis confirmed the different distribution of the hemispheric asymmetry score in newborns < 30 weeks old. Here, we show electrophysiological evidence of the early functional right lateralisation for pitch processing (detection and discrimination) arising by 30 GWs, but not before, in preterm newborns despite the longer environmental sensorial experience of newborns < 30 GWs. Generally, these findings suggest that the earlier right structural maturation in foetal epochs seems to be paralleled by a right functional development.

To investigate the role of Lcl in adhesion and invasion, the experiment was repeated with Selleckchem Thiazovivin bacteria (5 × 107 bacteria mL−1) preincubated

with Lcl-specific antibodies (20 μg mL−1 bacteria culture) at 37 °C for 1 h before they were placed in contact with the eukaryotic cells. As a control, experiments were repeated with a xylanase C (XlnC) antibody. XlnC is a Streptomyces lividans secreted protein (Faury et al., 2004) and the XlnC antibodies were of the same isotype and produced under the same conditions as the Lcl-specific antibodies. Alternatively, for measuring adhesion to host cells, experiments were performed with immobilized purified, refolded Lcl protein. Lcl and BSA (negative control) were immobilized as films on flat-bottomed microtiter 96-well plates (Nunclon) at a concentration of 5 μg per well overnight at 4 °C. Films were blocked with 1% BSA, washed with phosphate-buffered saline (PBS), followed by addition of 100 μL of eukaryotic cell suspension (5 × 105 cells mL−1) to each well and incubation at room temperature for 1 h. Nonadherent cells were removed by two washes with PBS, and those that adhered to the films were stained with crystal violet. Plates were read at A595 nm. Additionally, the immobilized films were preincubated with Lcl-specific antibodies

(20, 2, PFT�� in vivo 0.2 μg per well) for 30 min on ice before adding the eukaryotic cells. Coimmunoprecipitation experiments were carried out using a host cell lysate in combination with refolded Lcl protein. First, pelleted A549 cells or macrophage-like cells were resuspended in solubilization buffer (150 mM NaCl, 50 mM Tris, pH 8.0, 0.2% Triton X-100) and sonicated. Samples (500 μL) of the lysate (0.5 μg μL−1) were incubated with refolded Lcl protein (10 μg in total) for 1 h at 4 °C, rotating end over end. Sepharose A powder (10 mg) was added to the 500 μL mixture and further rotated for 1 h at 4 °C, followed by centrifugation (5 min, Amylase 1000 g). The supernatant was subsequently incubated with Lcl-specific antibodies or complement component C1q receptor

(C1qR)-specific antibodies rotating for 1 h at 4 °C. This incubation step was followed by addition of 10 mg sepharose A powder again. After 1 h at 4 °C, the immunoprecipitates were isolated by centrifugation (5 min, 1000 g) and washed four times with 150 μL solubilization buffer. After resuspension in 2 × SDS loading dye, the samples were boiled and the immunoprecipitated proteins were visualized by immunodetection with Lcl-specific antibodies. As a control, samples containing only lysate and Lcl protein without antibodies and samples only containing antibodies were also incubated with the protein A sepharose powder. Statistical analyses were performed using the standard Student t-test with equal variances.

Previous reports have reported less consistent effects. One study found only ejaculate volume to be correlated with CD4 cell count, but sperm concentration and total sperm selleckchem count were lower in those men with CD4 count<200 cells/μL [14]. Two studies found CD4 cell count to correlate only with motility [12,17], while two others found CD4 cell count to positively correlate with motility and negatively correlate with abnormal morphology [13,15]. Although

the exact data were not presented, a further report demonstrated no effect of CD4 count on any parameter using a cut-off of 500 cells/μL [26]. An effect of CD4 cell count on these parameters is supported by studies reporting that a diagnosis of AIDS [11,15] and disease progression

[by Centers for Disease Control and Prevention (CDC) clinical categories [15] significantly affects spermatogenesis. VX-765 in vivo Unlike a report of a correlation between VL and type ‘b’ motility and sperm morphology [14] and another of a lower progressive motility in those with detectable VL [26], we found that VL had no effect on any parameter. Several small series reported no difference in any parameter in those taking antiretroviral medication [11–13,17,26], but many are hampered by small sample numbers. In contrast, we demonstrate that samples taken from men on HAART have significantly impaired sperm count, motility and morphology and a lower number of motile sperm available for use for insemination cycles post sperm washing. In view of the benefit of stable, well-controlled disease, as demonstrated by the relationship between CD4 cell count

and sperm parameters, it might have been expected that there would be a similar benefit of Urease undetectable VL. However, our data suggest that any such potential benefit is counterbalanced by the effect of commencing HAART. The effect of antiretrovirals remains difficult to separate from the effect of HIV infection, and few studies have prospectively assessed the effect of treatment. One report found that those on zidovudine treatment, regardless of disease stage, had parameters similar to those of untreated early disease stage patients [16]. One study assessed 26 men about to start treatment for 12 weeks, and reported an overall increase in sperm motility and normal morphology, with no effect on sperm count [27]. A case report of a sperm donor who seroconverted during the course of donation demonstrated a reduction in semen volume, sperm motility and percentage of spermatozoa with normal morphology following infection over a course of 18 months [28].

Many plastic processes employ ectoenzymes that may restore locally ‘juvenile’ environments

in addition to generating new signaling molecules from cell surface and ECM products. The window for this type of research has just been opened and new views on basically important and medically relevant mechanisms of brain plasticity will emerge. These might include a deeper understanding of mental disorders including anxiety disorders (Pizzorusso, 2009), as well as schizophrenia and affective disorders that generally develop after the closure of major critical Selleck GSK3 inhibitor periods for higher brain functions of the prefrontal cortex after adolescence. We wish to thank Dr Amin Derouiche, Bonn, for providing a photomicrograph for Fig. 1. Research in the authors’ laboratories on this topic is funded by the DFG (GU230/5-1,2,3; HE3604/2-1) and by ERA-Net NEURON (Moddifsyn).

Abbreviations AMPAR AMPA receptor CSPG chondroitin sulfate proteoglycan ECM extracellular matrix ECS extracellular space MMP matrix metalloprotease PNN perineuronal net tPA tissue-type NVP-BKM120 in vitro plasminogen activator “
“Although it is accepted that new neurons continue to be generated in the hippocampal dentate gyrus (DG) throughout adulthood, it has recently become apparent that this process is not homogeneous, and that a small region of the DG lacks neurogenesis. Here, we show that the relative area of this neurogenesis quiescent zone (NQZ) did not vary

after the peak in hippocampal postnatal neurogenesis and until animals reached adulthood, although the ratio between its actual volume and the total volume of the DG doubled during this time. However, we were able to identify a few mitotic cells that reside within this subregion in early adolescent rats. Furthermore, these cells can be activated, and 1 week of voluntary exercise was enough to significantly increase the number of mitotic cells within the NQZ of adolescent rats. There was, however, no corresponding increase in the number of new neurons in this subregion of the DG, suggesting that some factor necessary to allow these Fossariinae cells to develop into a mature phenotype is missing. Moreover, the same intervention was ineffective in increasing either proliferation or neurogenesis in older adult rats. Surprisingly, we found no evidence for the existence of an NQZ in the mouse DG, suggesting that the neurogenic process in these two rodent species is differently regulated. Understanding the molecular mechanisms underlying the existence of the NQZ in the rat DG might shed light on the processes that regulate adult neurogenesis and its modulation by factors such as aging and exercise. “
“A selection of influential FEMS publications to celebrate the 40th anniversary of FEMS.

The challenge of M. bovis was substituted with PBS in the control groups. Cells were resuspended in Trizol (Invitrogen) after 3 h of stimulation and stored at −80 °C. RNA was isolated from MDMs from the treatment and the control groups, according to the manufacturer’s protocol (Invitrogen), and then stored in RNase-free water at −80 °C. Total RNA was reverse transcribed to cDNA using the RevertAid first-strand cDNA synthesis Kit (Fermentas, Lithuania). For animal samples, expression levels in eight genes (seven selected genes and one control) were examined with real-time PCR. The H3 histone family 3A gene (H3F3A) was used as a control gene for BIBW2992 animal samples to normalize

expression data for target genes (MacHugh et al.,

2009). Gene expression levels were detected using the DNA Engine Opticon TM2 fluorescence detection system (MJ Research Inc.) and SYBR Green (RealMasterMix, Tiangen). The specific gene primer pairs are shown in Table 1. Real-time quantitative PCR data were analyzed using the method, and differences between groups were analyzed with a t-test by spss software. Cells were collected at various time points (3, 12 and 24 h) to prepare a cell layer smear. The cell smear was stained using the acid-fast staining method according to the Ziehl–Neelsen stain protocol. The intracellular M. bovis number count was performed using CFU assessment. Cells ABT-199 cell line from each timepoint (3, 12 and 24 h) were washed three times with PBS to remove the extracellular bacteria. Cells were then lysed with a 0.1% Triton X-100 solution, serially diluted in 7H9 medium with 0.05% Tween 80 and plated onto 7H10 agar plates containing 10% ADC. CFU were counted after incubation at 37 °C for 3–4 weeks. The gene expressions of IL1β, IL1A, IL1R1, TNF, TLR2, TLR4

and IL10 were examined by real-time PCR in MDMs in response to M. bovis stimulation from tuberculosis and healthy cattle (n=5 in each group). Of the seven genes examined in MDMs from tuberculosis animals, six genes (except IL1A) showed significant differential expression after 3 h of stimulation with M. bovis as compared with nonstimulated controls at the P≤0.05 level (Fig. 1, Table S2). IL1, Rucaparib mw IL1R1 and TNF-α genes showed increased expression 3-h poststimulation in both groups, which shows that the proinflammatory cytokine TNF-α, IL1 and its receptor IL1R1 play a role in the early interaction of host cells and M. bovis. Increased expression of TLR2 and TLR4 genes (2.64-fold and 6.49-fold) was also noted. These genes regulate the innate immune system. Anti-inflammatory cytokine IL-10 showed increased expression by 8.74-fold over the nonstimulated control. Of the seven genes from MDMs from healthy control animals, six genes (except IL1A) showed significant differential expression after 3 h of stimulation with M. bovis as compared with nonstimulated controls at the P≤0.05 level (Fig. 1, Table S3).

Baseline pretreatment values were used as a covariate for the evolution of values. Because this is a comparative observational study without a sample size calculation, all P-values were considered for descriptive purposes. All analyses were performed with spss v18 for Windows (SPSS Inc., Chicago, Wnt antagonist IL) and we considered a Type I error = 0.05. A total of 18 THAs indicated for the treatment of INFH were identified in 13 HIV-positive patients

(11 men and two women). Risk factors for HIV infection included sexual contact (n = 8; 62%), injecting drug use (IDU) (n = 4; 31%) and others (n = 1; 8%). At the time of HIV diagnosis, 67% of all patients were in stage C3, 11% in stage B3 and 22% in stage A2. The average duration of HIV infection prior to INFH diagnosis was 10 ± 6 years. The mean (± SD) duration of antiretroviral treatment at the time of INFH diagnosis was 9 ± 5 years. The most recent viral load within the 3 months prior to the intervention was <50 copies/ml in all patients, except for one case (1250 HIV-1 RNA copies/ml). The most recent CD4 T-lymphocyte count within the 3-month period prior to surgical intervention was (mean ± SD 434 ± 256 cells/μl (21 ± 10%). Patients had received treatment with a protease inhibitor (PI) for a mean (± SD) of 3.9 ± 2.7 years and with a nucleoside reverse transcriptase inhibitor (NRTI) for 8.1 ± 3.9 years. The control

group consisted of 36 THAs in 27 HIV-negative individuals (21 men and six women). The mean (± SD) age was 44.3 ± 9.1 years in the HIV-infected group Selleckchem Ponatinib and 47.0 ± 11.1 years in the control group (P = 0.45). The right/left hip ratio was 12/6 in the HIV-infected group and 15/17 in the control group (P = 0.15). The mean (± SD) duration of the follow-up period was 3.3 ± 2.5 years in the HIV-infected group and 5.5 ± 5.9 years in the control group (P = 0.08). All patients included in the study had at least 1 year of follow-up.

Table 1 shows comorbidities in each group. No differences were Olopatadine found with regard to body mass index or in the preanaesthetic assessment between the two groups. The frequency of chronic coinfection with hepatitis B virus (HBV) or hepatitis C virus (HCV) was significantly higher in the HIV-positive group. HIV-positive patients more often had antecedents of IDU and coinfection with HBV/HCV. In general, patients from the HIV-negative group presented with more comorbidities than those from the HIV-positive group. No significant differences were found in the time from the onset of initial symptoms to the diagnosis of INFH or in the INFH radiological state at the time of diagnosis (Table 2). Of the 18 THAs in the HIV-positive group, at the time of diagnosis, three were found to be in state I–II (17%) and 15 in state III–IV (83%). In the control group, eight were found to be in state I–II (22%) and 28 in state III–IV (78%) (P = 0.

The peak phases in three brain areas (OB, CPU and SN) differed slightly but significantly between the R-MAP and R-Water groups (interaction between brain area and treatment, F2,44 = 0.72, P = 0.49; main effect of treatment, F1,44 = 7.53, P = 0.009). In the SCN-lesioned rats, the peak phases in four brain areas (OB, CPU, PC and SN) were significantly different between the R-MAP and R-Water groups (interaction between brain area and treatment, F3,60 = 6.35, P = 8.3 × 10−4; main effect of treatment, F1,60 = 4.65, P = 0.035; selleck products Fig. 7C). A significant difference was revealed in the CPU and SN by a post hoc Fisher’s PLSD test (F7,60 = 8.05, P = 0.003 for CPU; P = 0.003 for SN). When compared between

the SCN-intact and SCN-lesioned rats (Fig. 7D), the peak phases in the three brain areas (OB, CPU and SN) were significantly different under R-MAP (interaction between brain area and SCN-lesion, F2,46 = 15.14, P = 8.9 × 10−6; main effect of SCN-lesion, F1,46 = 26.73, P = 5.0 × 10−6). A post hoc Fisher’s PLSD test revealed a significant

difference in the selleck screening library OB and SN (F5,46 = 12.26, P = 0.013 for OB; P = 8.0 × 10−9 for SN). Under R-Water (Fig. 7E), the peak phases in the four brain areas examined were significantly different between the SCN-intact and SCN-lesioned rats (interaction between brain area and SCN-lesion, F3,55 = 2.98, P = 0.039; main effect of SCN-lesion, F1,55 = 23.59, P = 1.0 × 10−5). A significant difference was revealed in the CPU and PC by a post hoc Fisher’s

PLSD test (F7,55 = 12.99, P = 4.2 × 10−5 for CPU; P = 0.010 for PC). The amplitude of first circadian peak in the SCN-intact rats (Fig. 8A) differed significantly among the four brain areas (effect of brain area, F3,60 = 54.19, P = 4.5 × 10−17) but not between the R-MAP and R-Water groups (interaction between brain area and treatment, F3,60 = 0.70, P = 0.56; main effect of treatment, F1,60 = 1.15, P = 0.29). The amplitude in the SCN-lesioned rats differed significantly among the four brain areas (effect of brain area, F3,61 = 17.81, P = 2.0 × 10−8; interaction between brain area and treatment, F3,61 = 3.43, P = 0.023; main effect of treatment, F1,61 = 3.99, P = 0.050). A post hoc Fisher’s PLSD test revealed a significant difference between the R-MAP and R-Water groups in the OB and PC (F7,61 = 9.67, CYTH4 P = 0.006 for OB; P = 0.031 for PC). When compared between the SCN-intact and SCN-lesioned rats, the amplitudes did not differ in the R-MAP group (interaction between brain area and SCN-lesion, F2,46 = 1.33, P = 0.28; main effect of SCN-lesion, F1,46 = 2.54, P = 0.12) but did significantly differ in the R-Water group (interaction between brain area and SCN-lesion, F3,55 = 15.86, P = 1.5 × 10−7; main effect of SCN-lesion, F1,55 = 14.00, P = 4.4 × 10−4).