Presence Of Gamma Research Articles

Laser surface cladding of CoNiCrAlY as Bond Coat on INCONEL718 substrate for thermal barrier coating application

This study delves into laser surface cladding of CoNiCrAlY onto Inconel718 employing a 6.6 kW continuous wave diode laser with varying process parameters. The resulting microstructure, microhardness, and isothermal oxidation resistance are meticulously assessed. Laser surface cladding yields a defect free microstructure with the presence of gamma (γ), gamma prime (γ’) and beta (β) phases. There is negligible dilution at the interface with a minimum HAZ. The average microhardness of the clad zone ranges from 425 to 465 HV, increasing with increase in scan speed and decrease in power. The high temperature oxidation resistance property of the clad surface is superior to the same developed by high velocity oxy fuel spraying.

Performance characteristics of a tungsten collimator and UVTRON flame sensor in the detection of alpha-induced radioluminescence

It has been established that the UVTRON flame sensor (Hamamatsu) can detect alpha-induced radioluminescence, but that the presence of gamma and beta radiation both interfere with this detection. A UVTRON was placed inside a tungsten collimator and exposed to a range of radioisotopes, 210Po, 241Am, 137Cs, 90Sr and 60Co, to investigate the effect of shielding the UVTRON. The collimator is a cylinder with a hole in the curved wall to allow light and particles to access the interior, without providing a direct shine path to the UVTRON sensor. Ultraviolet C (wavelength 180–280 nm) radioluminescence is reflected onto the UVTRON sensor using a UVC reflecting mirror. It was found that the collimator does not affect the low background count of the UVTRON, but that it does greatly reduce the UVC signal reaching the UVTRON from an alpha source. For example, the collimator reduced the signal by 94% at 60 mm, and 78% at 120 mm, where the signals were still far greater than the background (88 and 84 times background respectively). Beta particles entering the collimator, although not directly impacting on the UVTRON, do increase the count, likely due to bremsstrahlung radiation. The collimator attenuates gamma radiation dependent on the gamma energy, but as expected, does not block it. When using more than one source, the count is cumulative and therefore it may be possible to determine the presence of UVC radioluminescence through subtraction of the gamma and beta element of the signal. The results and findings of the experiments carried out are presented herewithin.

Microstructural change during laser welding of Inconel 718

In this study, Inconel 718 superalloy has been subjected to laser welding using a 2 kW continuous wave (CW) Yb-Fibre laser at a constant power of 1800 W with a varied scan speed of 1400 mm/min, 1200 mm/min, and 1000 mm/min, respectively. The effect of process parameters on microstructure, and microhardnes and nano-mechanical property has been studied. There is formation of reinforcement on the face of the weld zone and sometimes in the root. The weld reinforcement height has been found to vary from 51.3 μm to 72.9 μm and increases with increase in scan speed of laser beam. The microstructure of the weld zone is fine columnar in the solid liquid interface followed by dendritic region and with equiaxed morphology in the middle. In the inter-columnar, interdendritic or grain boundary regions there are presence of gamma and gamma prime precipitates. The orientation of grains in terms of micro-texture has been carefully analysed by EBSD analysis. The texture of the weld zone is marginally different from that of the base metal. There is a marginal decrease in microhardness (from 280 VHN for as-received to 246 VHN) in laser welded Inconel 718. A detailed nano-mechanical property of the weld zone has been studied by nano-hardness measurement.

Physicochemical Properties of Plukenetia volubilis L. Seeds and Oxidative Stability of Cold-pressed Oil (Green Nut Oil)

Cold-pressed oil from the seeds of Plukenetia volubilis L. (green nut oil, GNO) has been shown to be rich in n-3 fatty acid content, similar to flaxseed and perilla oils. These fatty acids are readily oxidized at high temperatures and under UV irradiation, limiting their potential applications. We therefore set out to clarify the physicochemical properties of green nuts. In addition, to explore means of the degradation process, we investigated the oxidation characteristics of GNO and the effects of oxidation on fatty acids. Results showed that GNO was stable up to 140 degrees C and exhibited greater UV resistance than the other oils. This may be related to the 10% lower alpha-linolenic acid content of GNO, compared to the other oils, as well as the presence of gamma- and delta-tocopherols. GNO was capable of tolerating a certain degree of heat processing.

MOMENTS OF NONIDENTICAL ORDER STATISTICS FROM BURR XII DISTRIBUTION WITH GAMMA AND NORMAL OUTLIERS

There are some distributions with no simple closed form for distribution functions such as the Normal and Gamma distributions. This will be the problem if we want to find moments of nonidentical order statistics in the presence of Gamma and Normal outliers observations. We used the idea of approximating Normal and Gamma distributions with Burr type XII distribution. We get single moments for order statistics from sample of independent nonidentically distributed Burr XII random variables that contains p-outlier from Normal or Gamma distributions. Approximating these distributions with Burr XII distribution and then we compared the results by previous method.

Design and performance tests of the calorimetric tract of a Compton Camera for small-animals imaging

The bio-distribution and targeting capability of pharmaceuticals may be assessed in small animals by imaging gamma-rays emitted from radio-isotope markers. Detectors that exploit the Compton concept allow higher gamma-ray efficiency compared to conventional Anger cameras employing collimators, and feature sub-millimeter spatial resolution and compact geometry. We are developing a Compton Camera that has to address several requirements: the high rates typical of the Compton concept; detection of gamma-rays of different energies that may range from 140 keV ( 99 m Tc) to 511 keV ( β + emitters); presence of gamma and beta radiation with energies up to 2 MeV in case of 188Re. The camera consists of a thin position-sensitive Tracker that scatters the gamma ray, and a second position-sensitive detection system to totally absorb the energy of the scattered photons (Calorimeter). In this paper we present the design and discuss the realization of the calorimetric tract, including the choice of scintillator crystal, pixel size, and detector geometry. Simulations of the gamma-ray trajectories from source to detectors have helped to assess the accuracy of the system and decide on camera design. Crystals of different materials, such as LaBr 3 GSO and YAP, and of different size, in continuous or segmented geometry, have been optically coupled to a multi-anode Hamamatsu H8500 detector, allowing measurements of spatial resolution and efficiency.

Immunocytochemical study by two photon fluorescence microscopy of the distribution of GABA(A) receptor subunits in rat cerebellar granule cells in culture.

An immunocytochemical investigation of the expression of alpha(1), alpha(6), beta(2/3), gamma(2) and delta subunits was performed on rat cerebellum granule cells in culture by the two photon microscopy technique. The first four subunits appear to be expressed abundantly in these cells, whereas the delta one seems to be expressed at a lower level. Another major difference in the distribution of these subunits is that whereas alpha(6), beta(2/3) and gamma(2) appear only on plasma membranes alpha(1) and delta are present mainly in the cell bodies cytoplasm. Still another difference was found in that the presence of gamma(2) on neurites is "polarized", preferentially labelling neurites with the appearance of dendrites. The subunits alpha(6) and beta(2/3) appear to label all types of neurites, with beta(2/3) being by far the most heavily expressed subunit type. A final distinct characteristic is that alpha(6) and, even more, gamma(2) appear to accumulate in the cytoplasmic domains immediately below the cone of emergence of neurites. This suggests a conspicuous transport of such subunits from the site of synthesis in the cell body to the site of final expression in the neurites (dendrites and axon terminals).

Kinetic modification of the alpha(1I) subunit-mediated T-type Ca(2+) channel by a human neuronal Ca(2+) channel gamma subunit.

1. Voltage-sensitive Ca(2+) channels (VSCCs) are often heteromultimeric complexes. The VSCC subtype specifically expressed by skeletal muscle has long been known to contain a gamma subunit, gamma(1), that is only expressed in this tissue. Recent work, initiated by the identification of the mutation present in the stargazer mouse, has led to the identification of a series of novel potential Ca(2+) channel gamma subunits expressed in the CNS. 2. Based on bioinformatic techniques we identified and cloned the human gamma(2), gamma(3) and gamma(4) subunits. 3. TaqMan analysis was used to quantitatively characterise the mRNA expression patterns of all the gamma subunits. All three subunits were extensively expressed in adult brain with overlapping but subunit-specific distributions. gamma(2) and gamma(3) were almost entirely restricted to the brain, but gamma(4) expression was seen in a broad range of peripheral tissues. 4. Using a myc epitope the gamma(2) subunit was tagged both intracellularly at the C-terminus and on a predicted extracellular site between the first and second transmembrane domains. The cellular distribution was then examined immunocytochemically, which indicated that a substantial proportion of the cellular pool of the gamma(2) subunit was present on the plasma membrane and provided initial evidence for the predicted transmembrane topology of the gamma subunits. 5. Using co-transfection techniques we investigated the functional effects of each of the gamma subunits on the biophysics of the T-type VSCC encoded by the alpha(1I) subunit. This revealed a substantially slowed rate of deactivation in the presence of gamma(2). In contrast, there was no significant corresponding effect of either gamma(3) or gamma(4) on alpha(1I) subunit-mediated currents.

Molecular and functional studies of the gamma subunit of the sodium pump.

This article reviews our studies of the gamma subunit of the sodium pump. Gamma is a member of the FXYD family of small, single transmembrane proteins and is expressed predominantly in the kidney tubule. There are two major variants of gamma which function similarly to bring about two distinct effects, one on K'(ATP) and the other, on K(K), the affinity of the pump for K+ acting as a competitor of cytoplasmic Na+. In this way, gamma is believed to provide a self-regulatory mechanism for maintaining the steady-state activity of the pump in the kidney. Our studies also suggest that K+ antagonism of cytoplasmic Na+ activation of the pump is relevant not only to the presence of gamma in the kidney, but probably some hitherto undefined factor(s) in other tissues, most notably heart. The interesting possibility that not only gamma but other members of the FXYD family regulate ion transport in a tissue-specific manner is discussed.

Nucleoside diphosphate kinase and Cl(-)-sensitive protein phosphorylation in apical membranes from ovine airway epithelium.

We have previously shown that nucleotide species (adenosine triphosphate [ATP] or guanosine triphosphate [GTP]), [Cl-], and anion species determine the steady-state phosphorylation of apical membrane proteins within human airway epithelium in vitro. We found that a Cl(-)-regulated 37-kD protein (p37) principally phosphorylated with GTP but not ATP as substrate. Here we show that apical membranes from sheep tracheal epithelium also contain a Cl(-)-regulated 37-kD phosphoprotein (p37s) and characterize one of the kinases involved in the regulation of p37s. Analysis of phosphorylation of apical membrane proteins with gamma[32P]GTP in the presence of MgCl2 showed that two proteins circa 19 and 21 kD (p19s and p21s) were transiently phosphorylated before p37s. Renaturation of apical membrane proteins within polyacrylamide gels showed that p19s and p21s autophosphorylated with either gamma[32P]GTP or gamma[32P]ATP as substrates, suggesting that the two proteins were kinases. Immunoblotting and immunoprecipitation with a specific polyclonal antibody showed that p21s was a membrane-bound isoform of nucleoside diphosphate kinase (NDPK, EC 2.7.4.6), a protein kinase which catalyzes transfer of terminal phosphate from ATP to diphosphate nucleotides and is, among other functions, essential for cell secretion. Incubation of apical membrane proteins in the presence of gamma[32P]ATP and guanosine diphosphate (GDP) (but not GDPbetaS) resulted in enhancement of phosphorylation of p37s. Dephosphorylation of NDPK was stimulated by the addition of Mg2+, Mn2+, and Co2+ (but not Zn2+ or Ca2+). Our data show that ovine trachea is a good model for further characterization of the chloride-dependent cascade in airway epithelium.

Angiotensin II induces activation of phosphatidylinositol 3-kinase in cardiomyocytes.

Phosphatidylinositol 3-kinase phosphorylates membrane lipids at the third position of the inositol ring producing phosphoinositides, not on the pathway for production of 1,4,5-triphosphate. To test the hypotheses that angiotensin II (Ang II) activates phosphatidylinositol 3-kinase in cardiomyocytes and that this pathway is involved in Ang II-induced protein synthesis. Cardiomyocytes, in culture, from 7-day-old chick embryonic hearts were treated with Ang II and the activation of phosphatidylinositol 3-kinase was assessed after immunoprecipitation with antibodies to the p85 subunit of phosphatidylinositol 3-kinase by the conversion of PI (phosphatidylinositol) to phosphatidylinositol 3-monophosphate (PIP) in the presence of gamma-[32P]-ATP and analyzed by thin-layer chromatography. Western blotting was performed after antiphosphotyrosine immunoprecipitation with antibodies to the p85 subunit of phosphatidylinositol 3-kinase. Protein synthesis was assessed by [35S]-methionine incorporation and polyacrylamide gel electrophoresis. Ang II stimulated phosphatidylinositol 3-kinase activity dramatically, with 4.5- and 3.5-fold increases in PIP formation after 1 and 5 min, respectively. The involvement of tyrosine kinases was demonstrated by Western blotting in which Ang II increased tyrosine phosphorylation of a protein recognized by antibodies to the 85 kDa subunit of phosphatidylinositol 3-kinase. Furthermore, the tyrosine kinase inhibitor lavendustin A blocked Ang II-stimulated phosphatidylinositol 3-kinase activity and conversion of phosphatidylinositol to PIP. Ang II increased new protein synthesis as reflected by the significantly (P < 0.05) greater incorporation of [35S]-methionine into cardiomyocytes treated with Ang II. The link between Ang II and protein synthesis was mediated in part through phosphatidylinositol 3-kinase because the phosphatidylinositol 3-kinase inhibitor wortmannin blocked the effect of Ang II on protein synthesis. Increased production both of nuclear and of cytosolic proteins was demonstrated by agarose gel electrophoresis of these cellular components of Ang II-treated cardiomyocytes. Wortmannin produced a general inhibition of the synthesis of nuclear and cytosolic proteins, with a greater effect on nuclear proteins. The action of wortmannin on nuclear protein synthesis was confirmed by similar findings with another phosphatidylinositol 3-kinase inhibitor, LY294002. Phosphatidylinositol 3-kinase activation by Ang II occurs through a pathway utilizing tyrosine phosphorylation. Furthermore, this pathway is involved in cardiomyocyte protein synthesis and the possibility that it is operative in Ang II-mediated cardiac hypertrophy arises.

Subcellular localization of the GABAA receptor gamma 2 subunit in the rat spinal cord.

The fine subcellular organization of the GABAA receptor complex in the adult rat spinal ventral horn was analysed by immunocytochemistry using a specific polyclonal antiserum raised against the gamma 2 subunit. This subunit confers benzodiazepine sensitivity on the chloride channel of the GABAA receptor. With both fluorescent and peroxidase staining, the immunoreactivity was mainly observed in the grey matter and more specifically in the dorsal and ventral horns on medium and large neurons. A high number of immunostained somata were clustered in regions corresponding to motor nuclei. On the neuronal surface, labelling appeared as fluorescent dots over the more diffuse staining that was present on the soma and proximal part of dendrites. At the ultrastructural level, peroxidase end product was in most cases associated with the internal side of postsynaptic differentiations facing terminal boutons enriched with pleiomorphic small clear vesicles. The positively stained synapses were encountered on proximal dendrites of neurons and throughout the neuropil of the ventral horn (layers VII-IX). An immunoreactivity on the postsynaptic membrane was occasionally found to decorate large pieces of membrane not directly apposed to presynaptic active zones. In addition, presynaptic labelling was observed at axoaxonic contacts and at extrasynaptic sites on membranes within boutons, sometimes themselves apposed to gamma 2 immunoreactivity. Finally, we also observed gamma 2 immunoreactivity at the cytosolic face of the plasma membrane of some glial elements. These results give morphological evidence for the involvement of GABAA receptors in both post- and presynaptic inhibition in the rat spinal ventral horn. The presence of gamma 2 subunit immunoreactivity at these different synaptic contacts suggests that the two types of inhibition can be modulated by benzodiazepine drugs. The findings also provide anatomical evidence for the possible regulation of GABA release through an autoreceptor, and for GABAergic communication between neuronal and glial components.

Cyclic AMP-regulated AChR assembly is independent of AChR subunit phosphorylation by PKA.

Forskolin treatment of cells expressing Torpedo acetylcholine receptors leads to enhanced assembly efficiency of subunits, which correlates with increased phosphorylation of the gamma subunit. To determine the role of the two potential protein kinase A sites of the gamma subunit in receptor assembly, cell lines expressing different mutant receptors were established. Mouse fibroblast cell lines stably expressing wild-type Torpedo acetylcholine receptor alpha, beta, delta subunits plus one of three gamma subunit mutations (S353A, S354A, or S353,354A) were established to identify the protein kinase A phosphorylation sites of gamma in vivo, and to determine if increased phosphorylation of the gamma subunit leads to enhanced expression of receptors. We found that both serines (353, 354) in gamma are phosphorylated in vivo by protein kinase A, however, phosphorylation of either or both of these sites does not lead to increased assembly efficiency. We established a cell line expressing alpha, beta, and gamma(S353,354A) subunits only (no delta), and found that the presence of delta (or its phosphorylation) is also not necessary for the observed stimulation by forskolin. alpha beta gamma, alpha gamma, and beta gamma associations were stimulated by forskolin but alpha beta and alpha delta interactions were not. These data imply that the presence of gamma is necessary for forskolin action. We postulate that forskolin may stimulate acetylcholine receptor expression through a cellular protein that is involved in the folding and/or assembly of protein complexes, and that forskolin may regulate the action of such a protein through phosphorylation.

The third gamma subunit of the gamma-aminobutyric acid type A receptor family.

Cloned cDNAs encoding a member of the gamma-aminobutyric acid type A receptor gamma-subunit class were isolated from rat-brain-mRNA-derived libraries. The gamma 3 mRNA is present in cortex, claustrum, caudate putamen, and some thalamic nuclei, particularly the medial geniculate nucleus, where it is the predominant gamma-subunit transcript. The gamma 3 gene is expressed at very low levels in cerebellum and hippocampus. In coexpression experiments with the alpha 1 and beta 2 subunits, gamma 3 imparts benzodiazepine binding to gamma-aminobutyric acid type A receptors and forms gamma-aminobutyric acid-gated benzodiazepine-modulated chloride channels that exhibit a larger conductance than alpha 1 beta 2 receptor channels. Furthermore, the presence of gamma 3 in place of gamma 2 in alpha 1 beta 2 gamma x receptors generates a marked decrease in the affinity of agonists while leaving the affinity of antagonists or negative modulators largely unaffected.

ATP mediated receptor interconversion as a model of estrogen action: Isolation of the factor which converts the non-estrogen binding form of the receptor to the lower affinity binding form

Two steroid binding states of an estrogen receptor each with different equilibrium constants ( K d values) R x ( K d = 0.06 nM) and R y , ( K d = 0.8 nM) have been identified and characterized in the hen and estrogen-stimulated chick oviduct. A third non-estrogen binding form of the receptor, designated R nb , has also been identified. These three forms of the receptor are interconvertible and appear to have a common molecular weight of approx. 66,000 under denaturing conditions. Hydroxytamoxifen binds preferentially and with high affinity to R x ( K d 0.03 nM) and the conversion of R x to R y which is mediated by gamma phosphoryl group of ATP is also inhibitable by hydroxytamoxifen. Thus receptor interconversion, which may have general application to hormone action, potentially explains agonist/antagonist activity. The conversion of the non-estrogen binding form of the receptor ( R nb ) to the lower affinity receptor ( R y ) in chick oviduct cytosol is catalyzed by a reaction requiring the loss of the terminal phosphoryl moiety from ATP. There is a specific requirement for Mg 2+. We now describe that ammonium sulfate fractionation of the cytosol allows the separation of the receptor entities from the “activating factor” ( F y ) that catalyzes the conversion of R nb to R y In the presence of gamma [ 32P]-ATP at 30°C the purified non-steroid binding form of the receptor is phosphorylated. Phosphoamino acid analysis using Partisil-10 SAX anion exchange resin demonstrates that a serine is phosphorylated; and quantitation of the phosphorylation is indicative of one phosphoserine/receptor molecule. Treatment of the receptor with the partially purified activating factor to induce estradiol binding causes a dramatic reduction in phosphorylation.

Determination of gamma glutamyltransferase in completely haemolysed blood samples.

A simple method for the determination of gamma glutamyltransferase (GGT) in completely haemolysed blood samples is described. Haemolysed blood was incubated in the presence of gamma glutamyl-p-nitroanilide and glycylglycine at 37 degrees C for 15 min, and the enzyme reaction was terminated by adding trichloroacetic acid. The pH in the supernatant was adjusted to 7.5 by using a buffer, and the amount of p-nitroaniline formed was measured spectrophotometrically. The correlation between this method and the GGT activity in serum as determined by the standard kinetic method was good, r = 0.982. The blood GGT activity (y) was related to the serum activity (x) as follows: y = 0.415x + 3.8. The effect of storage of blood at room temperature on the GGT activity was studied. The enzyme activity did not change during the first 12 days of storage.

Phosphorylation of vesicular stomatitis virus proteins as a possible contributing factor in virion uncoating.

The relationship between the in vitro phosphorylation of vesicular stomatitis virus (VSV) proteins and virion uncoating was examined. Activation of the VSV virion kinase with low concentrations of melittin, the active peptide component of bee venom, in the presence of gamma-[32P] ATP resulted in the phosphorylation of virion proteins. Following the in vitro phosphorylation of VSV proteins in the presence of melittin and deoxyadenosine triphosphate, the virion envelope was disrupted based on the accessibility of the internal ribonucleoprotein core (RNP) to the heavy metal stain, uranyl acetate, as determined by electron microscopic observation. The RNP structure was not observed in unphosphorylated virions treated with melittin and uranyl acetate. Phosphorylated virions treated with uranyl acetate subsequently lost the capacity for transcription whereas unphosphorylated virions treated with the stain retained transcriptase activity. These observations suggest that phosphorylation of VSV proteins may contribute to virion uncoating by disrupting the virus envelope.