Tonically Active Kainate Receptors (tKARs) : A Novel Mechanism Regulating Neuronal Function in the Brain

Fast excitatory transmission between neurons in the central nervous system is mainly mediated by L-glutamate acting on ligand gated (ionotropic) receptors. These are further categorized according to their pharmacological properties to AMPA (2-amino-3-(5-methyl-3-oxo-1,2- oxazol-4-yl)propanoic acid), NMDA (N-Methyl-D-aspartic acid) and kainate (KAR) subclasses. In the rat and the mouse hippocampus, development of glutamatergic transmission is most dynamic during the first postnatal weeks. This coincides with the declining developmental expression of the GluK1 subunit-containing KARs. However, the function of KARs during early development of the brain is poorly understood. The present study reveals novel types of tonically active KARs (hereafter referred to as tKARs) which play a central role in functional development of the hippocampal CA3-CA1 network. The study shows for the first time how concomitant pre- and postsynaptic KAR function contributes to development of CA3-CA1 circuitry by regulating transmitter release and interneuron excitability. Moreover, the tKAR-dependent regulation of transmitter release provides a novel mechanism for silencing and unsilencing early synapses and thus shaping the early synaptic connectivity. The role of GluK1-containing KARs was studied in area CA3 of the neonatal hippocampus. The data demonstrate that presynaptic KARs in excitatory synapses to both pyramidal cells and interneurons are tonically activated by ambient glutamate and that they regulate glutamate release differentially, depending on target cell type. At synapses to pyramidal cells these tKARs inhibit glutamate release in a G-protein dependent manner but in contrast, at synapses to interneurons, tKARs facilitate glutamate release. On the network level these mechanisms act together upregulating activity of GABAergic microcircuits and promoting endogenous hippocampal network oscillations. By virtue of this, tKARs are likely to have an instrumental role in the functional development of the hippocampal circuitry. The next step was to investigate the role of GluK1 -containing receptors in the regulation of interneuron excitability. The spontaneous firing of interneurons in the CA3 stratum lucidum is markedly decreased during development. The shift involves tKARs that inhibit medium-duration afterhyperpolarization (mAHP) in these neurons during the first postnatal week. This promotes burst spiking of interneurons and thereby increases GABAergic activity in the network synergistically with the tKAR-mediated facilitation of their excitatory drive. During development the amplitude of evoked medium afterhyperpolarizing current (ImAHP) is dramatically increased due to decoupling tKAR activation and ImAHP modulation. These changes take place at the same time when the endogeneous network oscillations disappear. These tKAR-driven mechanisms in the CA3 area regulate both GABAergic and glutamatergic transmission and thus gate the feedforward excitatory drive to the area CA1. Here presynaptic tKARs to CA1 pyramidal cells suppress glutamate release and enable strong facilitation in response to high-frequency input. Therefore, CA1 synapses are finely tuned to high-frequency transmission; an activity pattern that is common in neonatal CA3-CA1 circuitry both in vivo and in vitro. The tKAR-regulated release probability acts as a novel presynaptic silencing mechanism that can be unsilenced in response to Hebbian activity. The present results shed new light on the mechanisms modulating the early network activity that paves the way for oscillations lying behind cognitive tasks such as learning and memory. Kainate receptor antagonists are already being developed for therapeutic use for instance against pain and migraine. Because of these modulatory actions, tKARs also represent an attractive candidate for therapeutic treatment of developmentally related complications such as learning disabilities.

Boundary layer swirling flow through a conical hydrocyclone

The paper deals with the flow and heat-transfer problem of a steady axisymmetric laminar incompressible boundary layer swirling flow of a fluid through a conical hydrocyclone. The implicit finitedifference scheme is used to solve the partial differential equations governing the flow. The effect of swirl is found to be more pronounced on the longitudinal skin friction than on the tangential skin friction and heat transfer. The skin friction and heat transfer increase with swirl or with longitudinal distance. Swirl also gives rise to velocity overshoot in the longitudinal velocity profiles and the magnitude of the velocity overshoot increases as the swirl parameter increases. The results are found to be in good agreement with those of the local nonsimilarity and momentum integral methods but they differ appreciably from those of the local similarity method except for the longitudinal skin friction which is fairly in good agreement with that of the local similarity method.Die Arbeit beschäftigt sich mit der Strömung und dem Wärmeübergang in einem konischen Zyklon unter der Voraussetzung stationärer, achsensymmetrischer, laminarer, inkompressibler Grenzschichtströmung. Ein implizites Differenzenverfahren wird benutzt, um die partiellen Differentialgleichungen zu lösen. Der Einfluß des Dralls ist besonders ausgeprägt auf die longitudinale Komponente der Oberflächenreibung, weniger dagegen bei der tangentialen Komponente und beim Wärmeübergang. Die Oberflächenreibung und der Wärmeübergang nehmen zu mit dem Drall, sowie mit dem longitudinalen Abstand. Der Drall erzeugt ein Überschießen der Geschwindigkeit in der longitudinalen Abstand. Der Drall erzeugt ein Überschießen der Geschwindigkeit in der Längsrichtung. Die Größe des Überschusses nimmt mit wachsendem Drallparameter zu. Die Resultate stimmen gut mit den Ergebnissen der Theorie der lokalen Nichtähnlichkeit und der Impulsintegralmethode überein. Dagegen weichen sie mit Ausnahme der longitudinalen Komponente der Oberflächenreibung beträchtlich von den Resultaten der Theorie der lokalen Ähnlichkeit ab.

Molecular genetics of Usher syndrome -inherited deafness and blindness

Usher syndrome (USH) is an inherited blindness and deafness disorder with variable vestibular dysfunction. The syndrome is divided into three subtypes according to the progression and severity of clinical symptoms. The gene mutated in Usher syndrome type 3 (USH3), clarin 1 (CLRN1), was identified in Finland in 2001 and two mutations were identified in Finnish patients at that time. Prior to this thesis study, the two CLRN1 gene mutations were the only USH mutations identified in Finnish USH patients. To further clarify the Finnish USH mutation spectrum, all nine USH genes were studied. Seven mutations were identified: one was a previously known mutation in CLRN1, four were novel mutations in myosin VIIa (MYO7A) and two were a novel and a previously known mutation in usherin (USH2A). Another aim of this thesis research was to further study the structure and function of the CLRN1 gene, and to clarify the effects of mutations on protein function. The search for new splice variants resulted in the identification of eight novel splice variants in addition to the three splice variants that were already known prior to this study. Studies of the possible promoter regions for these splice variants showed the most active region included the 1000 bases upstream of the translation start site in the first exon of the main three exon splice variant. The 232 aa CLRN1 protein encoded by the main (three-exon) splice variant was transported to the plasma membrane when expressed in cultured cells. Western blot studies suggested that CLRN1 forms dimers and multimers. The CLRN1 mutant proteins studied were retained in the endoplasmic reticulum (ER) and some of the USH3 mutations caused CLRN1 to be unstable. During this study, two novel CLRN1 sequence alterations were identified and their pathogenicity was studied with cell culture protein expression. Previous studies with mice had shown that Clrn1 is expressed in mouse cochlear hair cells and spiral ganglion cells, but the expression profile in mouse retina remained unknown. The Clrn1 knockout mice display cochlear cell disruption/death, but do not have a retinal phenotype. The zebrafish, Danio rerio, clrn1 was found to be expressed in hair cells associated with hearing and balance. Clrn1 expression was also found in the inner nuclear layer (INL), photoreceptor layer and retinal pigment epithelium layer (RPE) of the zebrafish retina. When Clrn1 production was knocked down with injected morpholino oligonucleotides (MO) targeting Clrn1 translation or correct splicing, the zebrafish larvae showed symptoms similar to USH3 patients. These larvae had balance/hearing problems and reduced response to visual stimuli. The knowledge this thesis research has provided about the mutations in USH genes and the Finnish USH mutation spectrum are important in USH patient diagnostics. The extended information about the structure and function of CLRN1 is a step further in exploring USH3 pathogenesis caused by mutated CLRN1 as well as a step in finding a cure for the disease.

Dielectric relaxation in bismuth layer-structured BaBi4Ti4O15 ferroelectric ceramics

The dielectric properties of BaBi4Ti4O15 ceramics were investigated as a function of frequency (10(2)-10(6) Hz) at various temperatures (30 degrees C-470 degrees C), covering the phase transition temperature. Two different conduction mechanisms were obtained by fitting the complex impedance data to Cole-Cole equation. The grain and grain boundary resistivities were found to follow the Arrhenius law associated with activation energies: E-g similar to 1.12 eV below T-m and E-g similar to 0.70 eV above T-m for the grain conduction; and E-gb similar to 0.93 eV below T-m and E-gb similar to 0.71 eV above T-m for the grain boundary conduction. Relaxation times extracted using imaginary part of complex impedance Z `'(omega) and modulus M `'(omega) were also found to follow the Arrhenius law and showed an anomaly around the phase transition temperature. The frequency dependence of conductivity was interpreted in terms of the jump relaxation model and was fitted to the double power law. (C) 2010 Elsevier B. V. All rights reserved.

Selective-Area Atomic Layer Deposition

Atomic layer deposition (ALD) is a method to deposit thin films from gaseous precursors to the substrate layer-by-layer so that the film thickness can be tailored with atomic layer accuracy. Film tailoring is even further emphasized with selective-area ALD which enables the film growth to be controlled also on the substrate surface. Selective-area ALD allows the decrease of a process steps in preparing thin film devices. This can be of a great technological importance when the ALD films become into wider use in different applications. Selective-area ALD can be achieved by passivation or activation of a surface. In this work ALD growth was prevented by octadecyltrimethoxysilane, octadecyltrichlorosilane and 1-dodecanethiol SAMs, and by PMMA (polymethyl methacrylate) and PVP (poly(vinyl pyrrolidone) polymer films. SAMs were prepared from vapor phase and by microcontact printing, and polymer films were spin coated. Microcontact printing created patterned SAMs at once. The SAMs prepared from vapor phase and the polymer mask layers were patterned by UV lithography or lift-off process so that after preparation of a continuous mask layer selected areas of them were removed. On these areas the ALD film was deposited selectively. SAMs and polymer films prevented the growth in several ALD processes such as iridium, ruthenium, platinum, TiO2 and polyimide so that the ALD films did grow only on areas without SAM or polymer mask layer. PMMA and PVP films also protected the surface against Al2O3 and ZrO2 growth. Activation of the surface for ALD of ruthenium was achieved by preparing a RuOX layer by microcontact printing. At low temperatures the RuCp2-O2 process nucleated only on this oxidative activation layer but not on bare silicon.

Temperature-Sensitive Src-Transformed Mdck Cells nn 3d Cultures as a Model of Malignant Transformation of Epithelial Cells

The equilibrium between cell proliferation, differentiation, and apoptosis is crucial for maintaining homeostasis in epithelial tissues. In order for the epithelium to function properly, individual cells must gain normal structural and functional polarity. The junctional proteins have an important role both in binding the cells together and in taking part in cell signaling. Cadherins form adherens junctions. Cadherins initiate the polarization process by first recognizing and binding the neighboring cells together, and then guiding the formation of tight junctions. Tight junctions form a barrier in dividing the plasma membranes to apical and basolateral membrane domains. In glandular tissues, single layered and polarized epithelium is folded into tubes or spheres, in which the basal side of the epithelial layer faces the outer basal membrane, and the apical side the lumen. In carcinogenesis, the differentiated architecture of an epithelial layer is disrupted. Filling of the luminal space is a hallmark of early epithelial tumors in tubular and glandular structures. In order for the transformed tumor cells to populate the lumen, enhanced proliferation as well as inhibition of apoptosis is required. Most advances in cancer biology have been achieved by using two-dimensional (2D) cell culture models, in which the cells are cultured on flat surfaces as monolayers. However, the 2D cultures are limited in their capacity to recapitulate the structural and functional features of tubular structures and to represent cell growth and differentiation in vivo. The development of three-dimensional (3D) cell culture methods enables the cells to grow and to be studied in a more natural environment. Despite the wide use of 2D cell culture models and the development of novel 3D culture methods, it is not clear how the change of the dimensionality of culture conditions alters the polarization and transformation process and the molecular mechanisms behind them. Src is a well-known oncogene. It is found in focal and adherens junctions of cultured cells. Active src disrupts cell-cell junctions and interferes with cell-matrix binding. It promotes cell motility and survival. Src transformation in 2D disrupts adherens junctions and the fibroblastic phenotype of the cells. In 3D, the adherens junctions are weakened, and in glandular structures, the lumen is filled with nonpolarized vital cells. Madin-Darby canine kidney (MDCK) cells are an epithelial cell type commonly used as a model for cell polarization. Its-src-transformed variants are useful model systems for analyzing the changes in cell morphology, and they play a role in src-induced malignant transformation. This study investigates src-transformed cells in 3D cell cultures as a model for malignant transformation. The following questions were posed. Firstly: What is the role of the composition and stiffness of the extracellular matrix (ECM) on the polarization and transformation of ts v-src MDCK cells in 3D cell cultures? Secondly: How do the culture conditions affect gene expression? What is the effect of v-src transformation in 2D and in 3D cell models? How does the shift from 2D to 3D affect cell polarity and gene expression? Thirdly: What is the role of survivin and its regulator phosphatase and tensin homolog protein (PTEN) in cell polarization and transformation, and in determining cell fate? How does their expression correlate with impaired mitochondrial function in transformed cells? In order to answer the above questions, novel methods of culturing and monitoring cells had to be created: novel 3D methods of culturing epithelial cells were engineered, enabling real time monitoring of a polarization and transformation process, and functional testing of 3D cell cultures. Novel 3D cell culture models and imaging techniques were created for the study. Attention was focused especially on confocal microscopy and live-cell imaging. Src-transformation disturbed the polarization of the epithelium by disrupting cell adhesion, and sensitized the cells to their environment. With active src, the morphology of the cell cluster depended on the composition and stiffness of the matrix. Gene expression studies revealed a broader impact of src transformation than mere continuous activity of src-kinase. In 2D cultures, src transformation altered the expression of immunological, actin cytoskeleton and extracellular matrix (ECM). In 3D, the genes regulating cell division, inhibition of apoptosis, cell metabolism, mitochondrial function, actin cytoskeleton and mechano-sensing proteins were altered. Surprisingly, changing the culture conditions from 2D to 3D affected also gene expression considerably. The microarray hit survivin, an inhibitor of apoptosis, played a crucial role in the survival and proliferation of src-transformed cells.

A Multi frequency study of Radio Intensity Variations For Active Galactic Nuclei of Different Optical Classes

A study of radio intensity variations at seven frequencies in the range 0.3 to 90 GHz for compact extragalactic radio sources classified as BL Lacs and high- and low-optical polarization quasars (HPQs and LPQs) is presented. This include the results of flux-density monitoring of 33 compact sources for three years at 327 MHz with the Ooty Synthesis Radio Telescope. The degrees of 'short-term' (tau less than about 1 yr) variability for the three optical types are found to be indistinguishable at low frequencies (less than 1 GHz), pointing to an extrinsic origin for the low-frequency variability. At high frequencies, a distinct dependence on optical type is present, the variability increasing from LPQs, through HPQs to BL Lacs. This trend persists even when only sources with ultra-flat radio spectra (alpha greater than -0.2) are considered. Implications of this for the phenomenon of high-frequency variability and the proposed unification schemes for different optical types of active galactic nuclei are discussed.

Atmospheric boundary-layer across Hadley and Ferrel cells over the Indian Ocean

A Southern Ocean Pilot cruise covering the latitudes from 10 degrees N to 56 degrees S in the open Indian Ocean was carried out during January February 2004. Surface and upper air data collected during this cruise are reported here. It is shown that the broad features of the atmosphere, in particular that of temperature, follow the tropical and mid-latitude weather expected during January February in this region. However, the atmospheric boundary-layer shows large variations, both in its height and structure between tropics and high latitudes. Strong influence of the surface heat flux on boundary layer structure is clearly seen. Humidity field reveals several local maxima and minima, suggesting a laminated atmosphere with air from different sources moving almost unmixed in adjacent layers.

On the possibility of an alpha-sq omega-type dynamo in a thin layer inside the sun

If the solar dynamo operates in a thin layer of 10,000-km thickness at the interface between the convection zone and the radiative core, using the facts that the dynamo should have a period of 22 years and a half-wavelength of 40 deg in the theta-direction, it is possible to impose restrictions on the values which various dynamo parameters are allowed to have. It is pointed out that the dynamo should be of alpha-sq omega nature, and kinematical calculations are presented for free dynamo waves and for dynamos in thin rectangular slabs with appropriate boundary conditions. An alpha-sq omega dynamo is expected to produce a significant poloidal field which does not leak to the solar surface. It is found that the turbulent diffusity eta and alpha-coefficient are restricted to values within about a factor of 10, the median values being eta of about 10 to the 10th sq cm/sec and alpha of about 10 cm/sec. On the basis of mixing length theory, it is pointed out that such values imply a reasonable turbulent velocity of the order 30 m/s, but rather small turbulent length scales like 300 km.

A Novel Method for the Uniform Incorporation of Grain-Boundary Layer Modifiers in Positive Temperature Coefficient of Resistivity Barium Titanate

The presently developed two-stage process involves diping the prefired porous disks of n-BaTiO3 in nonaqueous solutions containing Al-buty rate, Ti-isopropoxide, and tetraethyl silicate and subsequent sintering. This leads to uniform distribution of the grain-boundary layer (GBL) modifiers (Al2O3+ TiO2+ SiO2) and better control of the grain size as well as the positive temperature coefficient of resistivity characteristics. The technique is particularly suited for GBL modifiers in low concentrations (< 1%).

A sufficient criterion for Kelvin–Helmholtz instability in the magnetopause boundary‐layer region

The Kelvin–Helmholtz instability has been investigated for the magnetopause boundary‐layer region by the linearized method. The plasma in magnetosheath and magnetopause is assumed to be semi‐infinitely extended homogeneous, nondissipative, and incompressible. It is observed that, if one relation of two plasma speeds on the two sides of the magnetopause, wave number, and boundary‐layer thickness exceeds a certain threshold, the instability sets in. This new analytically sufficient criterion for excitation of instability in the three‐layer plasma flow generalizes the corresponding Chandrasekhar’s instability criterion for two‐layer plasma flow. The known results have been recovered and modified, the new results have been discovered. It is proved that the velocity threshold for the onset of instability is low when the magnitude of the magnetosheath and boundary‐layer region magnetic field and the angle between them are small. Also the threshold depends on the direction of plasma flow. The following results are observed numerically. The growth of the instability is sensitive to the magnetic field direction in the magnetosheath. A slight variation in the magnetic field direction in the second region can substantially change the relative velocity threshold for instability. When the ratio of the density of the second and third layer (magnetosphere) increases or that of the first and third layer decreases, the threshold decreases. Apart from this a necessary criterion for instability is obtained for a particular case.

Broadband stacked three-layer circular microstrip antenna arrays

The broadband aspects of stacked three-layer electromagnetically coupled circular microstrip antenna arrays are investigated experimentally. Experiments carried out on 8-element linear microstrip antenna arrays, using optimized stacked three-layer circular microstrip antenna elements, configured in E- and H-planes, have exhibited an impedance bandwidth of 20 percent, with a high gain and a good pattern shape with sidelobe as well as crosspolarization levels better than -20 dB through a scan angle of 40 deg from the broadside.