A new method for modulating the activity of parvalbumin-positive neurons and cerebellar Purkinje cells in vivo

Neurons can be divided into various classes according to their location, morphology, neurochemical identity and electrical properties. They form complex interconnected networks with precise roles for each cell type. GABAergic neurons expressing the calcium-binding protein parvalbumin (Pv) are mainly interneurons, which serve a coordinating function. Pv-cells modulate the activity of principal cells with high temporal precision. Abnormalities of Pv-interneuron activity in cortical areas have been linked to neuropsychiatric illnesses such as schizophrenia. Cerebellar Purkinje cells are known to be central to motor learning. They are the sole output from the layered cerebellar cortex to deep cerebellar nuclei. There are still many open questions about the precise role of Pv-neurons and Purkinje cells, many of which could be answered if one could achieve rapid, reversible cell-type specific modulation of the activity of these neurons and observe the subsequent changes at the whole-animal level. The aim of these studies was to develop a novel method for the modulation of Pv-neurons and Purkinje cells in vivo and to use this method to investigate the significance of inhibition in these neuronal types with a variety of behavioral experiments in addition to tissue autoradiography, electrophysiology and immunohistochemistry. The GABA(A) receptor γ2 subunit was ablated from Pv-neurons and Purkinje cells in four separate mouse lines. Pv-Δγ2 mice had wide-ranging behavioral alterations and increased GABA-insensitive binding indicative of an altered GABA(A) receptor composition, particularly in midbrain areas. PC-Δγ2 mice experienced little or no motor impairment despite the lack of inhibition in Purkinje cells. In Pv-Δγ2-partial rescue mice, a reversal of motor and cognitive deficits was observed in addition to restoration of the wild-type γ2F77 subunit to the reticular nucleus of thalamus and the cerebellar molecular layer. In PC-Δγ2-swap mice, zolpidem sensitivity was restored to Purkinje cells and the administration of systemic zolpidem evoked a transient motor impairment. On the basis of these results, it is concluded that this new method of cell-type specific modulation is a feasible way to modulate the activity of selected neuronal types. The importance of Purkinje cells to motor control supports previous studies, and the crucial involvement of Pv-neurons in a range of behavioral modalities is confirmed.

Stress-strain characteristics of Tress-strain characteristics of reinforcing steel bars under cyclic loading

This paper presents test results for 22 high strength deformed bars and nine mild steel bars subjected to monotonic repeated and reversed axial loading to determine the stress-strain behavior. Equations have been proposed for the stress-strain curves and have been compared with test results. Satisfactory agreement was obtained.

Processing map for hot working of alpha-zirconium

The hot deformation characteristics of alpha-zirconium in the temperature range of 650 °C to 850 °C and in the strain-rate range of 10-3 to 102 s-1 are studied with the help of a power dissipation map developed on the basis of the Dynamic Materials Model.[7,8,9] The processing map describes the variation of the efficiency of power dissipation (η =2m/m + 1) calculated on the basis of the strain-rate sensitivity parameter (m), which partitions power dissipation between thermal and microstructural means. The processing map reveals a domain of dynamic recrystallization in the range of 730 °C to 850 °C and 10−2 to 1−1 with its peak efficiency of 40 pct at 800 °C and 0.1 s-1 which may be considered as optimum hot-working parameters. The characteristics of dynamic recrystallization are similar to those of static recrystallization regarding the sigmoidal variation of grain size (or hardness) with temperature, although the dynamic recrystallization temperature is much higher. When deformed at 650 °C and 10-3 s-1 texture-induced dynamic recovery occurred, while at strain rates higher than 1 s-1, alpha-zirconium exhibits microstructural instabilities in the form of localized shear bands which are to be avoided in processing.

Stress-Strain Characteristics of Reinforcing Steel Bars Under Cyclic Loading

This paper presents test results for 22 high strength deformed bars and nine mild steel bars subjected to monotonic repeated and reversed axial loading to determine the stress-strain behavior. Equations have been proposed for the stress-strain curves and have been compared with test results. Satisfactory agreement was obtained.

Studies on ß-arylhydrazone-imine nickel(II) complexes—Examples of metal template syntheses

ß-arylhydrazone-imine ligand complexes of nickel(II), namely, 4,10-dimethyl-5,9-diazatrideca-4,9-diene-2,12-dione-3,11-diphenylhydrazonato nickel(II), Ni(acacpn)(N2Ph-R)2 and 1,11-diphenyl-3,9-dimethyl-4,8-diazaun-deca-3,8-diene,1,11-dione-2,10-diphenyl hydrazonato nickel(II), Ni (beacpn) (N2Ph-R)2, [R = H, o-CH3p-CH3] have been prepared by metal template reactions and characterized. Both the azomethine nitrogens and α-nitrogens of bis-hydrazone form the coordination sites of the square-planar geometry around the nickel(II) ion. Loss of CO from the molecule and subsequently an interesting methyl group migration to the nucleus of the chelate ring have been observed in the mass spectrum. Structures are proposed based on the spectral and magnetic properties.

On characterization of deformation microstructure in Boron modified Ti-6Al-4V alloy

Addition of boron in small quantities to various titanium alloys have shown significant improvement in mechanical behavior of materials. In the present study, electron back-scatter diffraction (EBSD) techniques have been applied to investigate the deformation microstructure evolution in boron modified two-phase titanium alloy Ti-6Al-4V. The alloy was hot compressed at 750 degrees C up to 50% height reduction at two different strain rates (10(-3) s(-1) and 1 s(-1)). The EBSD analyses indicated significant differences in deformed microstructure of the base alloy and the alloy containing boron. A strong subgrain formation tendency was observed along with inhomogeneous distribution of dislocations inside large a colonies of Ti64. In contrast, a colonies were relatively strain free for Ti64 + B, with more uniform dislocation density distribution. The observed difference is attributed to microstructural modifications viz, grain size refinement and presence of TiB particles at grain boundary produced due to boron addition. (C) 2010 Elsevier B.V. All rights reserved.

Microstructural features of flow instability in α-titanium cylinders under high strain rate compression at 25°C to 400°C

Cylindrical specimens of commercial pure titanium have been compressed at strain rates in the range of 0.1 to 100 s-1 and temperatures in the range of 25-degrees-C to 400-degrees-C. At strain rates of 10 and 100 s-1, the specimens exhibited adiabatic shear bands. At lower strain rates, the material deformed in an inhomogeneous fashion. These material-related instabilities are examined in the light of the ''phenomenological model'' and the ''dynamic materials mode.'' It is found that the regime of adiabatic shear band formation is predicted by the phenomenological model, while the dynamic materials model is able to predict the inhomogeneous deformation zone. The criterion based on power partitioning is competent to predict the variations within the inhomogeneous deformation zone.

Processing map for hot working of stainless steel type AISI 316L

Processing and instability maps using a dynamic materials model have been developed for stainless steel type AISI 316L in the temperature range 600-1250-degrees-C and strain rate range 0.001-100 s-1 with a view to optimising its hot workability. Stainless steel type AISI 316L undergoes dynamic recrystallisation, with a peak efficiency of 35% at 1250-degrees-C and 0.05 s-1, which are the optimum parameters for hot working this material. The material undergoes dynamic recovery at 900-degrees-C and 0.001 s-1. The increase in the dynamic recrystallisation and dynamic recovery temperatures in comparison with stainless steel type AISI 304L is attributed to the presence of a backstress caused by the molybdenum additions. These results are in general agreement with those reported elsewhere on stainless steel type 316 deformed in hot extrusion and hot torsion. At temperatures < 850-degrees-C and strain rates > 10 s-1, the material exhibits flow localisation owing to adiabatic shear band formation, whereas at higher temperatures (> 850-degrees-C) and strain rates (> 10 s-1) mechanical twinning and wavy slip bands are observed. (C) 1993 The Institute of Materials.

Importance of COMT in the regulation of prefrontal dopamine

The prefrontal cortex (PFC), located in the anterior region of the frontal lobe, is considered to have several key roles in higher cognitive and executive functions. In general, the PFC can be seen as a coordinator of thought and action allowing subjects to behave in a goal-directed manner. Due to its anatomical connections with a variety of cortical and subcortical structures, several neurotransmitters, including dopamine, are involved in the regulation of PFC activity. In general, the majority of released dopamine is cleared by the dopamine transporter (DAT). In the PFC however, the number of presynaptic DAT is diminished, emphasizing the relative importance of catechol-O-methyltransferase (COMT) in dopamine metabolism. As a result, the role of COMT in the etiology of psychotic disorders is under constant debate. The present study investigated the role of COMT in prefrontal cortical dopamine metabolism by different neurochemical methods in COMT knockout (COMT-KO) mice. Pharmacological tools to inhibit other dopamine clearing mechanisms were also used for a more comprehensive and collective picture. In addition, this study investigated how a lack of the soluble (S-) COMT isoform affects the total COMT activity as well as the pharmacokinetics of orally administered L-dopa using mutant mice expressing only the membrane-bound (MB-) COMT isoform. Also the role of COMT in striatal and accumbal dopamine turnover during Δ9-tetrahydrocannabinol (THC) challenge was studied. We found markedly increased basal dopamine concentrations in the PFC, but not the striatum or nucleus accumbens (NAcc), of mice lacking COMT. Pharmacological inhibition of the noradrenaline transporter (NET) and monoamine oxidase (MAO) elevated prefrontal cortical dopamine levels several-fold, whereas inhibition of DAT did not. The lack of COMT doubled the dopamine raising effects of NET and MAO inhibition. No compensatory expression of either DAT or NET was found in the COMT-KO mice. The lack of S-COMT decreased the total COMT activity by 50-70 % and modified dopamine transmission and the pharmacokinetics of exogenous Ldopa in a sex and tissue specific manner. Finally, we found that subsequent tolcapone and THC increased dopamine levels in the NAcc, but not in the striatum. Conclusively, this study presents neurochemical evidence for the important role of COMT in the PFC and shows that COMT is responsible for about half of prefrontal cortical dopamine metabolism. This study also highlights the previously underestimated proportional role of MB-COMT and supports the clinical evidence of a gene x environment interaction between COMT and cannabis.

Excitation and correlation of N-14 overtone transitions and measurement of heteronuclear dipolar coupling using DAPT

For studying systems containing nitrogen, limited use of N-14 NMR spectroscopy has been made because of the large quadrupolar interaction experienced by the N-14 nucleus and the absence of a central transition. To overcome the above problem, use of overtone spectroscopy has been suggested. Though this approach has limited applicability for powder samples due to second order quadrupole broadening, it is useful for studying oriented samples and single crystals. Here, we demonstrate the use of the recently proposed dipolar assisted polarization transfer (DAPT) pulse scheme for exciting the overtone transitions. The pulse sequence may also be utilized as a two-dimensional experiment to obtain H-1-N-14 dipolar couplings and H-1 chemical shifts. (C) 2010 Elsevier B.V. All rights reserved.

Texture evolution and operative mechanisms during large-strain deformation of nanocrystalline nickel

The large-strain deformation of nanocrystalline nickel was investigated at room temperature and cryogenic (liquid N-2) temperature. Deformation mechanisms ranging from grain boundary sliding to slip, operate due to a wide distribution of grain sizes. These mechanisms leave their finger print in the deformation texture evolution during rolling of nanocrystalline nickel. The occurrence and severance of different mechanisms is understood by a thorough characterization of the deformed samples using X-ray diffraction, X-ray texture measurements, electron back-scattered diffraction and transmission electron microscopy. Crystal plasticity-based viscoplastic self-consistent simulations were used to further substantiate the experimental observations. Thus, a comprehensive understanding of deformation behavior of nanocrystalline nickel, which is characterized by simultaneous operation of dislocation-dominated and grain boundary-mediated mechanisms, has been developed.

X-ray diffraction line profile analysis of deformation microstructure in boron modified Ti-6Al-4V alloy

X-ray diffraction line profile analysis (XRDLPA) techniques have been applied to investigate the deformed microstructure of a recently developed boron modified two-phase titanium alloy Ti-6Al-4V. The alloy was hot compressed at 750 degrees C up to 50% height reduction at two different strain rates (10(-3) S-1 and 1 S-1). Microstructural parameters like average domain size, average microstrain within the domain and dislocation density of the two phases were determined using X-ray diffraction line profile analysis. The results indicate an increase in the microstrain and dislocation density for the alpha-phase and decrease for the beta-phase in the case of boron modified alloys as compared to the normal material. Microstructural modifications viz, the grain refinement and the presence of hard, brittle TiB particles in the case of boron modified alloy are held responsible for the observed difference in the dislocation density. (C) 2010 Elsevier Inc. All rights reserved.

The scratch hardness and friction of a soft rigid-plastic solid

The paper describes an experimental and analytical study of the normal and scratch hardnesses of a model soft rigid-plastic solid. The material known as ‘Plasticine’, a mixture of dry particles and a mineral oil, has been deformed with a range of rigid conical indentors with included angles of between 30° and 170°. The sliding velocity dependence of the computed scratch hardness and friction has been examined in the velocity range 0.19 mm/s to 7.3 m/s. Data are also described for the time dependence of the normal hardness and also the estimated rate dependence of the intrinsic flow stress. The latter values were estimated from data obtained during the upsetting of right cylinders. Three major conclusions are drawn from these data and the associated analysis. (1) A first-order account of the scratching force may be provided by adopting a model which sums the computed plastic deformation and interfacial sliding contributions to the total sliding work. This is tantamount to the adoption of the two-term non-interacting model of friction. (2) For this system during sliding, at high sliding velocities at least, the interface shear stress which defines the boundary condition is not directly related to the bulk shear stress. The interface rheological characteristics indicate an appreciable dependence on the imposed strain or strain rate. In particular, the relative contributions of the slip and stick boundary conditions appear to be a function of the imposed sliding velocity. (3) The computed normal and scratch hardness values are not simply interrelated primarily because of the evolving boundary conditions which appear to exist in the scratching experiments.

Dendritic Poly(ether imine) Based Gene Delivery Vector

The nonviral vector based gene delivery approach is attractive due to advantages associated with molecular-level modifications suitable for optimization of vector properties. In a new class of nonviral gene delivery systems, we herein report the potential of poly(ether Mine) (PETIM) dendrimers to mediate an effective gene delivery function. PETIM dendrimer, constituted with tertiary amine branch points, n-propyl ether linkers and primary amines at their peripheries, exhibits significantly reduced toxicities, over a broad concentration range. The dendrimer complexes pDNA effectively, protects DNA from endosomal damages, and delivers to the cell nucleus. Gene transfection studies, utilizing a reporter plasmid pEGFP-C1 and upon complexation with dendrimer, showed a robust expression of the encoded protein. The study shows that PETIM dendrimers are hitherto unknown novel gene delivery vectors, combining features of poly(ethylene imine)-based polymers and dendrimers, yet are relatively nontoxic and structurally precise.

Behaviour of the germ cell specific lamin through mammalian spermatogenesis as probed with monoclonal antibodies.

We had earlier identified a 60 kDa nuclear lamin protein (lamin(g)) unique to the germ cells of rat testis which was subsequently shown to be antigenically conserved in germ cells of grasshopper, rooster, frog and plants. We have now obtained eight monoclonal antibodies in mouse against this lamin(g) antigen. While all the eight Mabs reacted with lamin(g) antigen in an immunoblot analysis, only three Mabs (A(11)C(7), A(11)D(4), C1F7) showed strong reactivity in the immunofluorescence analysis of the germ cells. The Mabs A(11)C(7) and A(11)D(4) showed a slight cross-reactivity with rat liver lamin B. Indirect immunofluorescence analysis of pre-meiotic, meiotic and post-meiotic germ cells with Mabs have shown that while the lamin(g) is localized in the lamina structures of spermatogonia and round spermatids, it is localized to the phase dense regions of pachytene spermatocytes which is in conformity with our previous observations using rabbit polyclonal antibodies. The localization of the antigen in the germ cells was also confirmed by immunohistochemical staining of the thin sections of seminiferous tubules. By immunostaining the surface spread pachytene spermatocytes, the antigen was further localized to the telomeric ends of the paired homologous chromosomes. Using anti-somatic lamin B antibodies, we have also demonstrated the absence of somatic lamins in meiotic and post-meiotic germ cells. The lamina structure of pre-meiotic spermatogonial nucleus contains both somatic lamin B and lamin(g) as evidenced by immunofluorescence studies with two differently fluorochrome labelled anti-lamin B and anti-lamin(g) antibodies. The selective retention of lamin(g) in the pachytene spermatocytes is probably essential for anchoring the telomeric ends of the paired chromosomes to the inner nuclear membrane.