M2 receptors are required for spatiotemporal sequence learning in mouse primary visual cortex.

Acetylcholine is a neurotransmitter that plays a variety of roles in the central nervous system. It was previously shown that blocking muscarinic receptors with a nonselective antagonist prevents a form of experience-dependent plasticity termed "spatiotemporal sequence learning" in the mouse primary visual cortex (V1). Muscarinic signaling is a complex process involving the combined activities of five different G protein-coupled receptors, M1-M5, all of which are expressed in the murine brain but differ from each other functionally and in anatomical localization. Here we present electrophysiological evidence that M2, but not M1, receptors are required for spatiotemporal sequence learning in mouse V1. We show in male mice that M2 is highly expressed in the neuropil in V1, especially in thalamorecipient layer 4, and colocalizes with the soma in a subset of somatostatin-expressing neurons in deep layers. We also show that expression of M2 receptors is higher in the monocular region of V1 than it is in the binocular region but that the amount of experience-dependent sequence potentiation is similar in both regions and that blocking muscarinic signaling after visual stimulation does not prevent plasticity. This work establishes a new functional role for M2-type receptors in processing temporal information and demonstrates that monocular circuits are modified by experience in a manner similar to binocular circuits.NEW & NOTEWORTHY Muscarinic acetylcholine receptors are required for multiple forms of plasticity in the brain and support perceptual functions, but the precise role of the five subtypes (M1-M5) are unclear. Here we show that the M2 receptor is specifically required to encode experience-dependent representations of spatiotemporal relationships in both monocular and binocular regions of mouse V1. This work identifies a novel functional role for M2 receptors in coding temporal information into cortical circuits.

Expectation violations produce error signals in mouse V1.

Repeated exposure to visual sequences changes the form of evoked activity in the primary visual cortex (V1). Predictive coding theory provides a potential explanation for this, namely that plasticity shapes cortical circuits to encode spatiotemporal predictions and that subsequent responses are modulated by the degree to which actual inputs match these expectations. Here we use a recently developed statistical modeling technique called Model-Based Targeted Dimensionality Reduction (MbTDR) to study visually evoked dynamics in mouse V1 in the context of an experimental paradigm called "sequence learning." We report that evoked spiking activity changed significantly with training, in a manner generally consistent with the predictive coding framework. Neural responses to expected stimuli were suppressed in a late window (100-150 ms) after stimulus onset following training, whereas responses to novel stimuli were not. Substituting a novel stimulus for a familiar one led to increases in firing that persisted for at least 300 ms. Omitting predictable stimuli in trained animals also led to increased firing at the expected time of stimulus onset. Finally, we show that spiking data can be used to accurately decode time within the sequence. Our findings are consistent with the idea that plasticity in early visual circuits is involved in coding spatiotemporal information.

Temperature-Dependent Spin-Lattice Relaxation of the Nitrogen-Vacancy Spin Triplet in Diamond.

Spin-lattice relaxation within the nitrogen-vacancy (NV) center's electronic ground-state spin triplet limits its coherence times, and thereby impacts its performance in quantum applications. We report measurements of the relaxation rates on the NV center's |m_{s}=0⟩↔|m_{s}=±1⟩ and |m_{s}=-1⟩↔|m_{s}=+1⟩ transitions as a function of temperature from 9 to 474 K in high-purity samples. We show that the temperature dependencies of the rates are reproduced by an ab initio theory of Raman scattering due to second-order spin-phonon interactions, and we discuss the applicability of the theory to other spin systems. Using a novel analytical model based on these results, we suggest that the high-temperature behavior of NV spin-lattice relaxation is dominated by interactions with two groups of quasilocalized phonons centered at 68.2(17) and 167(12) meV.

Tracing exploitation of egg boons: an experimental study using fatty acids and stable isotopes.

Coordinated spawning of marine animals releases millions of planktonic eggs into the environment, known as egg boons. Eggs are rich in essential fatty acids and may be an important lipid subsidy to egg consumers. Our aim was to validate the application of fatty acid and stable isotope tracers of egg consumption to potential egg consumers and to confirm egg consumption by the selected species. We conducted feeding experiments with ctenophores, crustaceans and fishes. We fed these animals a common diet of Artemia or a commercial feed (Otohime) and simulated egg boons for half of them by intermittently supplementing the common diet with red drum (Sciaenops ocellatus) eggs for 10-94 days. Controls did not receive eggs. Fatty acid profiles of consumers fed eggs were significantly different from those of controls 24 h after the last egg-feeding event. Consumers took on fatty acid characteristics of eggs. In fishes and ctenophores, fatty acid markers of egg consumption did not persist 2-5 days after the last egg-feeding event, but markers of egg consumption persisted in crustaceans for at least 5-10 days. Additionally, consumption of eggs, which had high values of δ15N, led to δ15N enrichment in crustaceans and a fish. We conclude that fatty acids and nitrogen stable isotope can be used as biomarkers of recent egg consumption in marine animals, validating their use for assessing exploitation of egg boons in nature.

Sex and estrous cycle affect experience-dependent plasticity in mouse primary visual cortex.

Sex hormones can affect cellular physiology and modulate synaptic plasticity, but it is not always clear whether or how sex-dependent differences identified in vitro express themselves as functional dimorphisms in the brain. Historically, most experimental neuroscience has been conducted using only male animals and the literature is largely mute about whether including female mice in will introduce variability due to inherent sex differences or endogenous estrous cycles. Though this is beginning to change following an NIH directive that sex should be included as a factor in vertebrate research, the lack of information raises practical issues around how to design experimental controls and apply existing knowledge to more heterogeneous populations. Various lines of research suggest that visual processing can be affected by sex and estrous cycle stage. For these reasons, we performed a series of in vivo electrophysiological experiments to characterize baseline visual function and experience-dependent plasticity in the primary visual cortex (V1) of male and female mice. We find that sex and estrous stage have no statistically significant effect on baseline acuity measurements, but that both sex and estrous stage have can modulate two mechanistically distinct forms of experience dependent cortical plasticity. We also demonstrate that resulting variability can be largely controlled with appropriate normalizations. These findings suggest that V1 plasticity can be used for mechanistic studies focusing on how sex hormones effect experience dependent plasticity in the mammalian cortex.

Redox proteomics in aging rat brain: involvement of mitochondrial reduced glutathione status and mitochondrial protein oxidation in the aging process.

Increasing evidence supports the notion that increased oxidative stress is a fundamental cause in the aging process and in neurodegenerative diseases. As a result, a decline in cognitive function is generally associated with brain aging. Reactive oxygen species (ROS) are highly reactive intermediates, which can modify proteins, nucleic acids, and polyunsaturated fatty acids, leading to neuronal damage. Because proteins are major components of biological systems and play key roles in a variety of cellular functions, oxidative damage to proteins represents a primary event observed in aging and age-related neurodegenerative disorders. In the present study, with a redox proteomics approach, we identified mitochondrial oxidatively modified proteins as a function of brain aging, specifically in those brain regions, such as cortex and hippocampus, that are commonly affected by the aging process. In all brain regions examined, many of the identified proteins were energy-related, such as pyruvate kinase, ATP synthase, aldolase, creatine kinase, and α-enolase. These alterations were associated with significant changes in both cytosolic and mitochondrial redox status in all brain regions analyzed. Our finding is in line with current literature postulating that free radical damage and decreased energy production are characteristic hallmarks of the aging process. In additon, our results further contribute to identifying common pathological pathways involved both in aging and in neurodegenerative disease development.

Degradation of polycyclic aromatic hydrocarbons by Rigidoporus lignosus and its laccase in the presence of redox mediators.

The metabolism of polycyclic aromatic hydrocarbons (PAHs) was studied in vivo and in vitro in systems consisting of Rigidoporus lignosus and its laccase, in the presence of so-called "mediator" compounds. The static culture of the native fungal strain was able to metabolize anthracene and 2-methylanthracene, but not 9-nitroanthracene. The addition of redox mediators 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 1-hydroxybenzotriazole (HBT) or violuric acid (VA) led to a significant increase in the degradation of substrates. The oxidation of PAHs was not significant when purified laccase was used without the addition of mediators. The addition of these compounds increased the oxidation of all substrates by approximately 70-80% after 72 h of incubation. The degradation rate was highest for 2-methylanthracene in the presence of VA.

Comparison Between Folic Acid and gH625 Peptide-Based Functionalization of Fe3O4 Magnetic Nanoparticles for Enhanced Cell Internalization.

A versatile synthetic route based on magnetic Fe3O4 nanoparticle (MNP) prefunctionalization with a phosphonic acid monolayer has been used to covalently bind the gH625 peptide on the nanoparticle surface. gH625 is a membranotropic peptide capable of easily crossing the membranes of various cells including the typical human blood-brain barrier components. A similar synthetic route was used to prepare another class of MNPs having a functional coating based on PEG, rhodamine, and folic acid, a well-known target molecule, to compare the performance of the two cell-penetrating systems (i.e., gH625 and folic acid). Our results demonstrate that the uptake of gH625-decorated MNPs in immortalized human brain microvascular endothelial cells after 24 h is more evident compared to folic acid-functionalized MNPs as evidenced by confocal laser scanning microscopy. On the other hand, both functionalized systems proved capable of being internalized in a brain tumor cell line (i.e., glioblastoma A-172). These findings indicate that the functionalization of MNPs with gH625 improves their endothelial cell internalization, suggesting a viable strategy in designing functional nanostructures capable of first crossing the BBB and, then, of reaching specific tumor brain cells.

Effect of Nutritional Interventions on Micronutrient Status in Pregnant Malawian Women with Moderate Malnutrition: A Randomized, Controlled Trial.

Micronutrient deficiencies during pregnancy are common in Africa and can cause adverse outcomes. The objective was to measure micronutrient status and change in moderately malnourished pregnant Malawian women randomized to one of three nutritional interventions. Serum vitamin B12, 25-hydroxyvitamin D, folate, retinol, ferritin, zinc, albumin and C-reactive protein were measured in pregnant women with MUAC ≥20.6 cm and ≤23.0 cm at enrollment (n = 343) and after 10 weeks (n = 229) of receiving: (1) ready-to-use supplementary food (RUSF); (2) fortified corn-soy blend (CSB+) with multiple-micronutrient supplement (CSB+UNIMMAP); or (3) CSB+ with iron and folic acid (CSB+IFA). Each provided 100⁻300% Recommended Dietary Allowance of most micronutrients and 900 kcal/day. Birth length was measured in 272 infants. Enrollment measurements indicated deficiencies in vitamin B12 (20.9%) and zinc (22.3%), low values of ferritin (25.1%) and albumin (33.7%), and elevated CRP (46.0%). Vitamin B12 is known to decrease in the third trimester; the RUSF group had the smallest decrease from enrollment to week 10 (3%), compared to 20% decrease in the CSB+IFA group and 8% decrease in the CSB+UNIMMAP group (p = 0.001). Mean serum 25-hydroxyvitamin D increased most in the RUSF group (+6.4 ng/mL), compared to CSB+IFA (+1.7 ng/mL) and CSB+UNIMMAP (+2.7 ng/mL) (p < 0.001). Micronutrient deficiencies and inflammation are common among moderately malnourished pregnant women and had little improvement despite supplementation above the RDA, with the exception of vitamins B12 and D.

Multifunctional magnetic nanoparticles for enhanced intracellular drug transport.

In this paper we report the synthesis and characterization of biocompatible multi-functional magnetic nanoparticles (MNPs) able to enhance the intracellular transport of N-methylated drugs. The Fe3O4 magnetic core was first functionalized with a mixed monolayer consisting of two different phosphonic acids having terminal acetylenic and amino groups, which provide an active platform for further functionalization with organic molecules. Then, a tetraphosphonate cavitand receptor (Tiiii) bearing an azide moiety and the N-hydroxysuccinimide (NHS) activated forms of poly(ethylene glycol) (PEG), folic acid (FA) and carboxy-X-rhodamine (Rhod) were covalently anchored on alkyne and amine moieties respectively, through 1,3-dipolar cycloaddition and EDC/NHS coupling reactions. The obtained MNPs are biocompatible and possess magnetic, luminescence and recognition properties which make them suitable for multimodal theranostic applications. In particular, combined confocal microscopy and cytotoxicity experiments showed that these multi-functional MNPs are able to recognize a specific drug "in situ" and promote its cellular internalization, thus enhancing its efficiency.

Symptomatic cysts of the pineal gland: stereotactic diagnosis and treatment of two cases and review of the literature.

We report two cases of pineal gland cysts, both in young girls, causing aqueductal stenosis and symptomatic hydrocephalus, which were diagnosed and treated by Talairach's stereotactic method. This procedure permitted aspiration of the cyst content, normalization of aqueductal stenosis, and a differential histological diagnosis. Postoperative computed tomographic scan examination showed a stable reduction of the cystic volume and a complete clearance of the hydrocephalus. At present, the two patients are clinically normal and are leading a productive life (follow-up, 42 and 40 mo). It is important that this kind of nonneoplastic lesion be identified, in order to avoid confusion with pineal tumors and possible inappropriate treatment. Pineal cysts rarely become symptomatic. Up to the present, only 27 symptomatic cases have been described in the literature. Of those, signs of intracystic bleeding were found in 17 of the 21 cases in which the cyst contents were known. Moreover, in our own two cases, numerous hemosiderin-laden macrophages were present in the cystic fluid. We think, therefore, that intracystic bleeding has been the determining factor for the occurrence of symptoms in most of these cases. The authors underline the advantages of this type of stereotactic management, which can provide both diagnosis and treatment and thus avoid the possible risks of surgical excision and/or of radiotherapy.

Structure and stability of the insulin dimer investigated by molecular dynamics simulation.

Molecular dynamics simulation indicates that the dynamical behaviour of the insulin dimer is asymmetric. Atomic level knowledge of the interaction modes and protein conformation in the solvation state identifies dynamical structures, held by hydrogen bonds that stabilize, mainly in one monomer, the interaction between the chains. Dynamic cross-correlation analysis shows that the two insulin monomers behave asymmetrically and are almost independent. Solvation energy, calculated to evaluate the contribute of each interface residue to the dimer association pattern, well compares with the experimental association state found in protein mutants indicating that this parameter is an important factor to explain the association properties of mutated insulin dimers.

Calcified brain abscesses. Case report.

A surgically treated case of multiple contiguous brain abscesses at different stages of calcification is reported. The patient, a man 48 years old, had been suffering from epilepsy for 34 years and he was complaining of headaches and fever for 15 days when admitted to our clinic. The post-operative neurological recovery was good.

Spectroscopic and molecular dynamics simulation studies of the interaction of insulin with glucose.

The interaction between monomeric insulin and monosaccharides has been investigated through circular dichroism, fluorescence spectroscopy and two dimensional nuclear magnetic resonance. CD spectra indicate that D-glucose interacts with monomeric insulin whereas D-galactose, D-mannose and 2-deoxy-D-glucose have a lower effect. Fluorescence emission was quenched at sugar concentrations of 5-10 mM. Titration with the different sugars produces a quenching of the tyrosine spectrum from which a binding free energy value for the insulin-sugar complexes has been evaluated. Transfer nuclear Overhauser enhancement NMR experiments indicate the existence of dipolar interactions at short interatomic distances between C-1 proton of D-glucose in the beta form and the monomeric insulin. Further, NMR total correlation spectra experiments revealed that the hormone is in the monomeric form and that upon addition of glucose no aggregation occurs. The interaction does not involve relevant changes in the secondary structure of insulin suggesting that the interaction occur at the side chain level. Molecular dynamics simulations and modeling studies, based on the dynamic fluctuations of potential binding moiety sidechains, argued from results of NMR spectroscopy, provide additional informations to locate the putative binding sites of D-glucose to insulin.

Isolation and sequencing of a putative promoter region of the murine G protein beta 1 subunit (GNB1) gene.

The expression of the heterotrimeric GTP-binding protein beta 1 subunit gene (GNB1) is regulated by psychostimulants such as cocaine and amphetamines. Since the up-regulation appears to be one of the candidate processes of sensitization, it is necessary to elucidate the cellular and molecular mechanism of the GNB1 gene regulation for a better understanding the establishment of sensitization. In the present study, we describe the isolation and nucleotide sequence analysis of the GNB1 gene promoter region. We have isolated approximately 10 kb of the 5'-flanking region of the mouse of GNB1 gene and found potential elements involved in putative transcriptional control of the GNB1, such as AP1, AP2, Sp1, cyclic AMP response element, and nuclear factor kappa B recognition sites, within the sequences 0.3 kb upstream from the putative transcription start site. This region was highly rich in G + C content, but lacked TATA or CATT boxes. Comparing the nucleotide sequence of the cDNA clone with the human genome databases using the BLAST program a region containing putative exon 1 and promoter of the human GNB1 gene in chromosome 1 was found. The cloning and sequence analysis of an extensive portion of the 5'-flanking regulatory region of the GNB1 gene provides new insights into the factors involved in the regulation by psychostimulants of GNB1 expression.

[Experimental fetal hydrocephalus. Ventriculo-amniotic shunt]. / Hydrocéphalie foetale expérimentale. Shunt ventriculo-amniotique.

When the original idea was conceived 8 years ago to develop a surgical technique for the treatment of fetal hydrocephalus, the pathway towards prenatal surgery for this affection was totally unexplored. The operation appeared to be feasible because of the possibility of a ventriculo-amniotic shunt. The most important steps in our experimental studies in the ewe were creation of a fetal hydrocephalus by injection of a suspension of kaolin powder into cisterna magna; the treatment of the hydrocephalus by a mini-ventriculo-amniotic shunt, by cutting the cardiac end of a low pressure Pudenz pediatric catheter. Lambs were delivered by Cesarean two weeks before end of gestation. Coronal sections of brains of control and treated lambs showed reduction in hydrocephalus in the latter. These findings were the basis for the application of the method to treatment of human fetal hydrocephalus.

[Prevalence of non A - non B hepatitis in childhood (author's transl)]. / Incidenza di epatiti non A - non B in soggetti dell'età evolutiva.

Little is known from the literature about the epidemiology of non A - non B hepatitis (NANB/H) in childhood. Aim of this study was to assess the prevalence of NANB/H in a consecutive series of children with acute viral hepatitis hospitalized over an one year's period. Thirty children, 9 females, aged 3-12 years, were studied. Serial blood samples were tested for HBsAg, anti-HBs, anti-HBc, anti-HAV (Abbott RIA), anti-HAV-IgM (Absorption Staph. aureus protein A), anti-EBV (Immunofluorescence), anti-CMV, anti-Herpes s. virus (complement fixation). The diagnosis of NANB/H was based on the absence of these markers. Nineteen patients (63,3%) had type A, and 5 (16,6%), had type B hepatitis. One child showed antibodies anti-Herpes with rising titer and 5 (16.6%), 2 females, were considered suffering from NANB/H. None of these patients had been injected or haemotransfuded; all but one came from rural ambient and two from the same family. Two children had an anicteric course. The illness lasted less than 30 days in all but one, who showed three peaks of transaminases and recovered after 70 days. These data show a prevalence of NANB/H in childhood greater than that elsewhere reported, while the absence of injections suggests a way of infection other than parenteral.

Functionalization of PEGylated Fe3O4 magnetic nanoparticles with tetraphosphonate cavitand for biomedical application.

In this contribution, Fe3O4 magnetic nanoparticles (MNPs) have been functionalized with a tetraphosphonate cavitand receptor (Tiiii), capable of complexing N-monomethylated species with high selectivity, and polyethylene glycol (PEG) via click-chemistry. The grafting process is based on MNP pre-functionalization with a bifunctional phosphonic linker, 10-undecynylphosphonic acid, anchored on an iron surface through the phosphonic group. The Tiiii cavitand and the PEG modified with azide moieties have then been bonded to the resulting alkyne-functionalized MNPs through a "click" reaction. Each reaction step has been monitored by using X-ray photoelectron and FTIR spectroscopies. PEG and Tiiii functionalized MNPs have been able to load N-methyl ammonium salts such as the antitumor drug procarbazine hydrochloride and the neurotransmitter epinephrine hydrochloride and release them as free bases. In addition, the introduction of PEG moieties promoted biocompatibility of functionalized MNPs, thus allowing their use in biological environments.

Redox regulation of cellular stress response in multiple sclerosis.

Multiple sclerosis (MS) is an autoimmune-mediated neurodegenerative disease with characteristic foci of inflammatory demyelination in the brain, spinal cord, and optic nerves. Recent studies have demonstrated not only that axonal damage and neuronal loss are significant pathologic components of MS, but that this neuronal damage is thought to cause the permanent neurologic disability often seen in MS patients. Emerging finding suggests that altered redox homeostasis and increased oxidative stress, primarily implicated in the pathogenesis of MS, are a trigger for activation of a brain stress response. Relevant to maintenance of redox homeostasis, integrated mechanisms controlled by vitagenes operate in brain in preserving neuronal survival during stressful conditions. Vitagenes encode for heat shock proteins (Hsp) Hsp32, Hsp70, the thioredoxin and the sirtuin protein systems. In the present study we assess stress response mechanisms in the CSF, plasma and lymphocytes of control patients compared to MS patients. We found that the levels of vitagenes Hsp72, Hsc70, HO-1, as well as oxidative stress markers carbonyls and hydroxynonenals were significantly higher in the blood and CSF of MS patients than in control patients. In addition, an increased expression of Trx and sirtuin 1, together with a decrease in the expression of TrxR were observed. Our data strongly support a pivotal role for redox homeostasis disruption in the pathogenesis of MS and, consistently with the notion that new therapies that prevent neurodegeneration through nonimmunomodulatory mechanisms can have a tremendous potential to work synergistically with current MS therapies, unravel important targets for new cytoprotective strategies.