Neurotransmitter profile of saccadic omnipause neurons in nucleus raphe interpositus

Saccadic omnipause neurons (OPNs) are essential for the generation of saccadic eye movements. In primates OPNs are located near the midline within the nucleus raphe interpositus (rip). In the present study we used several different neuroanatomical methods to investigate the transmitters associated with OPNs in the monkey. Immunolabeling for the calcium-binding protein parvalbumin was employed to mark OPNs in the monkey and define the homologous cell group in cat and human. The use of antibodies against GABA, glycine (GLY), glutamate (GLU), serotonin (5-HT), and tyrosine hydroxylase revealed that the somata of OPNs are GLY immunoreactive, but they are devoid of GABA and 5-HT immunostaining. In situ hybridization with the GAD67 mRNA probe confirmed the negative GABA immunostaining of OPNs. 3H-GLY was injected into a projection field of OPNs, the rostral interstitial nucleus of the medial longitudinal fascicle (riMLF)--the vertical saccadic burst neuron area. This resulted in selective retrograde labeling of the OPNs in rip, while no labeling was found in the superior colliculus, which sends an excitatory projection to the riMLF. The somata and dendrites of putative burst neurons in the riMLF were contacted by numerous GLY- immunoreactive terminals. The quantitative analysis of immunoreactive terminal-like structures contacting OPNs revealed a strong input from GLY- and GABA-positive terminals on somata and dendrites, whereas GLU- positive puncta were mainly confined to the dendrites. Very few 5-HT and catecholaminergic terminals contacted OPN somata. Our findings suggest that OPNs use GLY as a neurotransmitter, and they receive numerous contacts from GABAergic, glycinergic, and glutaminergic afferents, and significantly fewer from monoaminergic inputs.</jats:p

Molecular aspects of insulin resistance, cell signaling pathways and breast cancer in relation to obesity.

A growing number of clinical studies validate a relation of insulin resistance and breast cancer in obese patients. We hypothesised that high plasma insulin levels cause aberrant insulin signalling in breast epithelial cells which may be responsible for an increase in cell proliferation, indicative of potential carcinogenesis and increased cancer progression. It was of particular interest to determine any differences of high insulin concentrations in activating the phosphoinositide-3 kinase (PI-3 kinase) pathway or the mitogen-activated protein kinase (MAP kinase) pathway, the latter being linked to increased cell proliferation. We used two cell line models to investigate the carcinogenic (MCF-10A, immortalised breast epithelial cells) and cancer progression (MDA-MB-231, ER-negative breast cancer cells) potential of insulin. Insulin treatment (100 nM, 24 h) increased cell proliferation in MCF-10A cells, but had no cell proliferative effect on MDA-MB-231 cells. Additionally expression of PCNA as marker of proliferation was tested. The use of PI-3 kinase and MAP kinase specific inhibitors (Wortmannin and PD98059, respectively) demonstrated both pathways being responsible for the observed increase in cell proliferation (MCF-10A). Simultaneous treatment with both inhibitors eliminated insulin induced cell proliferation entirely. Phosphorylation of ERK1/2 was examined as specific activity measurement of MAP kinase pathway. Insulin induced higher phosphorylation levels in MCF-10A cells than in MDA-MB-231. These preliminary results suggest that insulin may initiate carcinogenesis of breast epithelial cells by increasing cell proliferation rather than increasing cancer progression of existing tumours. These effects may be mediated by insulin activating both the PI-3 kinase and the MAP kinase signalling pathways

Molecular aspects linking insulin resistance to breast cancer by activation of cell signalling pathways.

Recent findings suggest a connection between obesity and breast cancer. Obesity is linked with higher incidences of insulin resistance as part of the metabolic syndrome, resulting in chronically elevated insulin plasma levels. We examined the effect of high insulin concentrations (100 nM) on estrogen-receptor (ER) negative breast cancer cells (MDA-MB-231) and normal breast epithelial cells (MCF-10a). Treatment with high insulin concentrations increased insulin receptor (IR) phosphorylation significantly in both cell lines. Phosphorylation of protein kinase B (Akt), representative of PI3-kinase cell signalling pathway activation was increased by 101% (p=0.0112) in MDA-MB-231 cells and by 81% (p=0.0031) in MCF-10a cells after 10 min insulin treatment. Phosphorylation of extracellular regulated kinase 1/2 (ERK1/2), representative of MAP-kinase cell signalling pathway activation did not change in both cell lines after 10 min of insulin treatment. Cell proliferation did not change in MDA-MB-231 cells and increased by 75% (p=0.0067) in MCF-10a cells after 24 h insulin treatment. Cell proliferation was decreased in MDA-MB-231 cells by 15% (p=0.0083) after 1 h treatment with PD98059, a MAP-kinase inhibitor. In MCF-10a cells cell proliferation was decreased by 51% (

Insulin-induced gene expression changes in breast cancer cells and normal breast epithelial cells.

Obesity increases breast cancer incidence rates in postmenopausal women. Chronic high levels of insulin, present in the majority of obese and insulin resistant patients, may provide the growth promoting stimulus to explain this connection. In this work, the cancer progression and cancer initiating properties of high insulin levels were examined in breast cancer cells (MDA-MB-231) and breast epithelial cells (MCF-10a), respectively. High insulin levels (100 nM) induced differential changes in cell proliferation in the two cell lines used. Human Cancer PathwayFinder DNA Microarrays (SABiosciences) were used to examine gene expression changes after insulin treatment. High insulin levels increased expression of genes involved in cell cycle control (e.g. cyclin D1) and DNA damage repair (e.g. ATM) in MDA-MB 231 cells and in MCF-10a cells (e.g. cyclin E1, CDC25a). Expression of genes responsible for mediating apoptosis and cell senescence (e.g. APAF, BAD, bcl-X) was decreased after insulin treatment in MDA-MB 231 cells but the expression of the same group of genes did not change in MCF-10a cells. High insulin levels increased expression of genes encoding for signal transduction molecules (e.g. AKT1) and transcription factors (e.g. FOS, JUN, MYC), and of genes responsible for invasion and metastasis (e.g. MMP2) in MCF-10a cells whereas gene expression of the same groups of genes did not change or was decreased in MDA-MB 231 cells. These results suggest a role for insulin resistance in breast cancer initiation and progression, aggravating the potential of breast cancer to evade apoptosis, to metastasise and may promote carcinogenesis of healthy epithelial cells

Stability of Ferromagnetism in Hubbard models with degenerate single-particle ground states

A Hubbard model with a N_d-fold degenerate single-particle ground state has ferromagnetic ground states if the number of electrons is less or equal to N_d. It is shown rigorously that the local stability of ferromagnetism in such a model implies global stability: The model has only ferromagnetic ground states, if there are no single spin-flip ground states. If the number of electrons is equal to N_d, it is well known that the ferromagnetic ground state is unique if and only if the single-particle density matrix is irreducible. We present a simplified proof for this result.Comment: accepted for publication in J. Phys.

Olive oil phenolics: effects on DNA oxidation and redox enzyme mRNA in prostate cells

J.L.Q. was supported by the University of Granada, Spain (Becas de Perfeccionamiento de Doctores Programme). D.K.S. was supported by a grant from World Cancer Research Fund (WCRF) and the other authors were supported by the Scottish Executive Rural and Agricultural Department (SERAD).Hydroxytyrosol, tyrosol and caffeic acid effects on hydrogen peroxide-induced DNA damage, hydroperoxide generation and redox enzyme gene expression were studied in oxidative-stress-sensitive human prostate cells (PC3). Hydroxytyrosol led to lower levels of hydroperoxides, DNA damage, and mRNA levels of classic glutathione peroxidase (GPx) for all the studied concentrations. Only hydroxytyrosol was effective at low concentrations (10 μM). TYROSOL REDUCED DNA OXIDATION ONLY AT HIGH (>50 Μm) concentrations and increased hydroperoxides, GPx and phospholipid hydroperoxide GPx mRNA levels. Caffeic acid elicited effects between those of the other two phenolics. Results indicate that hydroxytyrosol is the only significant antioxidant phenolic in olive oil and may be the major component accounting for its beneficial properties. Tyrosol appeared to exhibit pro-oxidant effects (only at high concentrations) and caffeic acid was neutral. Both number and position of hydroxyl groups appear to play a role in the cellular effects of hydroxytyrosol.University of Granada, Spain (Becas de Perfeccionamiento de Doctores Programme)World Cancer Research Fund (WCRF)Scottish Executive Rural and Agricultural Department (SERAD

Fuzzy cellular model for on-line traffic simulation

This paper introduces a fuzzy cellular model of road traffic that was intended for on-line applications in traffic control. The presented model uses fuzzy sets theory to deal with uncertainty of both input data and simulation results. Vehicles are modelled individually, thus various classes of them can be taken into consideration. In the proposed approach, all parameters of vehicles are described by means of fuzzy numbers. The model was implemented in a simulation of vehicles queue discharge process. Changes of the queue length were analysed in this experiment and compared to the results of NaSch cellular automata model.Comment: The original publication is available at http://www.springerlink.co

Magnetic structure in a U(Ru_0.92Rh_0.08)₂Si₂ single crystal studied by neutron diffraction in static magnetic fields up to 24 T

We report the high-field induced magnetic phase in single crystal of U(Ru0.92Rh0.08)2Si2. Our neutron study combined with high-field magnetization, shows that the magnetic phase above the first metamagnetic transition at Hc1 = 21.6 T has an uncompensated commensurate antiferromagnetic structure with propagation vector Q2 = ( 2/3 0 0) possessing two single-Q domains. U moments of 1.45 (9) muB directed along the c axis are arranged in an up-up-down sequence propagating along the a axis, in agreement with bulk measurements. The U magnetic form factor at high fields is consistent with both the U3+ and U4+ type. The low field short-range order that emerges from the pure URu2Si2 due to Rh-doping is initially strengthened by the field but disappears in the field-induced phase. The tetragonal symmetry is preserved across the transition but the a axis lattice parameter increases already at low fields. Our results are in agreement with itinerant electron model with 5f states forming bands pinned in the vicinity of the Fermi surface that is significantly reconstructed by the applied magnetic field.Comment: 5 pages, 4 figures, accepted as Rapid Communication, Physical Review B (2017