Adenosine Inhibits the Excitatory Synaptic Inputs to Basal Forebrain Cholinergic, GABAergic, and Parvalbumin Neurons in Mice

Coffee and tea contain the stimulants caffeine and theophylline. These compounds act as antagonists of adenosine receptors. Adenosine promotes sleep and its extracellular concentration rises in association with prolonged wakefulness, particularly in the basal forebrain (BF) region involved in activating the cerebral cortex. However, the effect of adenosine on identified BF neurons, especially non-cholinergic neurons, is incompletely understood. Here we used whole-cell patch-clamp recordings in mouse brain slices prepared from two validated transgenic mouse lines with fluorescent proteins expressed in GABAergic or parvalbumin (PV) neurons to determine the effect of adenosine. Whole-cell recordings were made from BF cholinergic neurons and from BF GABAergic and PV neurons with the size (>20 μm) and intrinsic membrane properties (prominent H-currents) corresponding to cortically projecting neurons. A brief (2 min) bath application of adenosine (100 μM) decreased the frequency but not the amplitude of spontaneous excitatory postsynaptic currents (EPSCs) in all groups of BF cholinergic, GABAergic, and PV neurons we recorded. In addition, adenosine decreased the frequency of miniature EPSCs in BF cholinergic neurons. Adenosine had no effect on the frequency of spontaneous inhibitory postsynaptic currents in cholinergic neurons or GABAergic neurons with large H-currents but reduced them in a group of GABAergic neurons with smaller H-currents. All effects of adenosine were blocked by a selective, adenosine A1 receptor antagonist, cyclopentyltheophylline (CPT, 1 μM). Adenosine had no postsynaptic effects. Taken together, our work suggests that adenosine promotes sleep by an A1 receptor-mediated inhibition of glutamatergic inputs to cortically projecting cholinergic and GABA/PV neurons. Conversely, caffeine and theophylline promote attentive wakefulness by inhibiting these A1 receptors in BF thereby promoting the high-frequency oscillations in the cortex required for attention and cognition

Localization of DNA methyltransferase-1 during oocyte differentiation, in vitro maturation and early embryonic development in cow

DNA methyltransferase-1 (Dnmt1) is involved in the maintenance of DNA methylation patterns and is crucial for normal mammalian development. The aim of the present study was to assess the localization of Dnmt1 in cow, during the latest phases of oocyte differentiation and during the early stages of segmentation. Dnmt1 expression and localization were assessed in oocytes according to the chromatin configuration, which in turn provides an important epigenetic mechanism for the control of global gene expression and represents a morphological marker of oocyte differentiation. We found that the initial chromatin condensation was accompanied by a slight increase in the level of global DNA methylation, as assessed by 5-methyl-cytosine immunostaining followed by laser scanning confocal microscopy analysis (LSCM). RT-PCR confirmed the presence of Dnmt1 transcripts throughout this phase of oocyte differentiation. Analogously, Dnmt1 immunodetection and LSCM indicated that the protein was always present and localized in the cytoplasm, regardless the chromatin configuration and the level of global DNA methylation. Moreover, our data indicate that while Dnmt1 is retained in the cytoplasm in metaphase II stage oocytes and zygotes, it enters the nuclei of 8-16 cell stage embryos. As suggested in mouse, the functional meaning of the presence of Dnmt1 in the bovine embryo nuclei could be the maintainement of the methylation pattern of imprinted genes. In conclusion, the present work provides useful elements for the study of Dnmt1 function during the late stage of oocyte differentiation, maturation and early embryonic development in mammals

Thermoreflectance investigation of the antiferromagnetic and paramagnetic phases of Cr

Thermoreflectance measurements have been performed on Cr single crystals at several temperatures above and below the Néel temperature. We observe dramatic changes induced by the magnetic phase transition. In contrast, static optical data fail to show appreciable differences in the (0.5-5.0)-eV photon-energy range. Magnetic ordering gives rise to the disappearance of transitions involving specific regions of the Fermi surface. New critical-point absorptions appear at the boundaries of the new Brillouin zone in antiferromagnetic Cr. Most of the observed experimental features have been identified by comparison with recent band-structure calculations

Electronic structure of Nb-Mo alloys

Thermoreflectance measurements on NbxMo1−x alloys (x=0.2, 0.5, 0.8) have been carried out in the 0.5-5.0 eV energy region. Augmented-plane-wave (APW) calculations for Nb at two different lattice parameters and for Mo, as well as coherent-potential-approximation calculations (CPA), have been carried out and have been used in the interpretation of the experimental results. Several optical transitions [Σ1(EF)→Σ3, G4(EF)→G1,N2→N′1], have been identified, and their concentration dependence followed. These results contribute significantly toward putting the interpretation of the optical properties of Nb, Mo, and their alloys on a much more secure footing. In particular, it has been confirmed that while the lower conduction bands behave roughly as predicted by the rigid-band model, the higher-lying conduction bands show distinctly non-rigid-band-like behavior

A study of the endogenous opioid system in the sudden infant death syndrome

To investigate the possible role of the endogenous opioid system in the pathogenesis of the sudden infant death syndrome (SIDS), we measured met-enkephalin immunoreactivity by radioimmunoassay in brain, lung, and adrenal glands of SIDS victims and of infants (controls) dying of nonneurologic causes. Met-enkephalin was stable in brain tissue up to 24 hours after death. On inspection, met-enkephalin levels in the cerebral cortex of SIDS victims were similar to those in controls. Levels in the caudate nucleus were lower in infants than in adults. In the medulla, the levels in SIDS cases and controls were not found to differ significantly. The linear relationship between the levels in the medulla and age was not detectably different in SIDS and controls. However, as a subset, levels in the control group significantly decreased with increasing age ( = 0.005), whereas levels in the SIDS group showed no correlation with age ( = 0.33). Levels of met-enkephalin in the adrenal gland of SIDS victims were similar to those in controls and were considerably lower than adult values. Lung tissue was assayed for beta-endorphin immunoreactivity and met-enkephalin: for both peptides the values in SIDS cases were low (femtomolar range) and similar to those in controls. These data suggest that met-enkephalin is not markedly overproduced in brains of SIDS victims. Future postmortem studies should focus on more subtle evidence of endogenous opioid overactivity such as differences in age related changes, receptor number, and levels of other endogenous opioid peptides.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25258/1/0000701.pd

In vitro evaluation of live attenuated vaccines against Salmonella enteritidis: humoral immune response

Salmonella enteritidis (SE) vaccination is one of the major tool to reduce the infection in commercial poultry. In this work, techniques, evaluating the presence of Ig-G and Ig-A in serum, tears and intestinal secretions, were studied to improve the knowledge of the humoral immune response following SE live attenuated vaccination. The Ig-G and Ig-A ELISA are important and easy tests to plan and optimize SE vaccination programs

Tracking the transfer of antimicrobial resistance genes from raw materials to sourdough breads

The present study hypothesizes that raw materials used in bread making can transfer antibiotic resistance genes (ARGs) to processed breads. Four types of flour and four types of semolina were purchased from supermarkets and inoculated with a commercial dried sourdough starter to make breads. The microbiological characteristics of all raw materials and fermented doughs were investigated. The levels of yeasts and lactic acid bacteria (LAB) increased up to 107 CFU/g. The values of pH decreased to 4.54–4.86 while total titratable acidity increased inversely. All unprocessed and processed samples, including breads, were analyzed by a molecular approach to detect bacterial and fungal DNAs and 17 antibiotic resistance genes for penicillins, macrolides, tetracyclines, and chloramphenicol. Illumina technology showed that the operational taxonomy units (OTUs) identified from unprocessed wheat milling products, fermented doughs, and baked products mainly belonged to Acetobacteraceae. Enterococci were present in all doughs. After baking, the relative abundance (RA)% of Enterococcus and Acetobacteraceae decreased. The DNA analyzed for fungal composition showed that Kazachstania humilis dominated dried sourdough starter and doughs, and its OTUs were also detected at high RA% in baked products. The search for ARGs revealed that all samples analyzed did not show resistance to penicillins, chloramphenicol, and macrolides. However, three of the semolinas included in this study (S1, S3 and S4) and the corresponding doughs (SD1, SD3 and SD4) were positive for tet(A) and tet(B) resistance genes. This work indicated that breads have a limited role in the dissemination of ARGs

Tracking the transfer of antimicrobial resistance genes from raw materials to sourdough breads

The present study hypothesizes that raw materials used in bread making can transfer antibiotic resistance genes (ARGs) to processed breads. Four types of flour and four types of semolina were purchased from supermarkets and inoculated with a commercial dried sourdough starter to make breads. The microbiological characteristics of all raw materials and fermented doughs were investigated. The levels of yeasts and lactic acid bacteria (LAB) increased up to 107 CFU/g. The values of pH decreased to 4.54–4.86 while total titratable acidity increased inversely. All unprocessed and processed samples, including breads, were analyzed by a molecular approach to detect bacterial and fungal DNAs and 17 antibiotic resistance genes for penicillins, macrolides, tetracyclines, and chloramphenicol. Illumina technology showed that the operational taxonomy units (OTUs) identified from unprocessed wheat milling products, fermented doughs, and baked products mainly belonged to Acetobacteraceae. Enterococci were present in all doughs. After baking, the relative abundance (RA)% of Enterococcus and Acetobacteraceae decreased. The DNA analyzed for fungal composition showed that Kazachstania humilis dominated dried sourdough starter and doughs, and its OTUs were also detected at high RA% in baked products. The search for ARGs revealed that all samples analyzed did not show resistance to penicillins, chloramphenicol, and macrolides. However, three of the semolinas included in this study (S1, S3 and S4) and the corresponding doughs (SD1, SD3 and SD4) were positive for tet(A) and tet(B) resistance genes. This work indicated that breads have a limited role in the dissemination of ARG