RESUMO
The perturbed free induction decay (PFID) observed in ultrafast infrared spectroscopy was used to unveil the rates at which different vibrational modes of the same atomic-scale defect can interact with their environment. The N_{3}VH^{0} defect in diamond provided a model system, allowing a comparison of stretch and bend vibrational modes within different crystal lattice environments. The observed bend mode (first overtone) exhibited dephasing times T_{2}=2.8(1) ps, while the fundamental stretch mode had surprisingly faster dynamics T_{2}<1.7 ps driven by its more direct perturbation of the crystal lattice, with increased phonon coupling. Further, at high defect concentrations the stretch mode's dephasing rate was enhanced. The ability to reliably measure T_{2} via PFID provides vital insights into how vibrational systems interact with their local environment.
RESUMO
The first experimental results from a new transmissive diagnostic instrument for synchrotron X-ray beamlines are presented. The instrument utilizes a single-crystal chemical-vapour-deposition diamond plate as the detector material, with graphitic wires embedded within the bulk diamond acting as electrodes. The resulting instrument is an all-carbon transmissive X-ray imaging detector. Within the instrument's transmissive aperture there is no surface metallization that could absorb X-rays, and no surface structures that could be damaged by exposure to synchrotron X-ray beams. The graphitic electrodes are fabricated inâ situ within the bulk diamond using a laser-writing technique. Two separate arrays of parallel graphitic wires are fabricated, running parallel to the diamond surface and perpendicular to each other, at two different depths within the diamond. One array of wires has a modulated bias voltage applied; the perpendicular array is a series of readout electrodes. X-rays passing through the detector generate charge carriers within the bulk diamond through photoionization, and these charge carriers travel to the nearest readout electrode under the influence of the modulated electrical bias. Each of the crossing points between perpendicular wires acts as an individual pixel. The simultaneous read-out of all pixels is achieved using a lock-in technique. The parallel wires within each array are separated by 50â µm, determining the pixel pitch. Readout is obtained at 100â Hz, and the resolution of the X-ray beam position measurement is 600â nm for a 180â µm size beam.
RESUMO
We demonstrate optical spin polarization of the neutrally charged silicon-vacancy defect in diamond (SiV^{0}), an S=1 defect which emits with a zero-phonon line at 946 nm. The spin polarization is found to be most efficient under resonant excitation, but nonzero at below-resonant energies. We measure an ensemble spin coherence time T_{2}>100 µs at low-temperature, and a spin relaxation limit of T_{1}>25 s. Optical spin-state initialization around 946 nm allows independent initialization of SiV^{0} and NV^{-} within the same optically addressed volume, and SiV^{0} emits within the telecoms down-conversion band to 1550 nm: when combined with its high Debye-Waller factor, our initial results suggest that SiV^{0} is a promising candidate for a long-range quantum communication technology.
RESUMO
Activation of an immune response requires intercellular contact between T lymphocytes and antigen-presenting cells (APC). Interaction of the T cell antigen receptor (TCR) with antigen in the context of major histocompatibility molecules mediates signal transduction, but T cell activation appears to require the induction of a second costimulatory signal transduction pathway. Recent studies suggest that interaction of CD28 with B7 on APC might deliver such a costimulatory signal. To investigate the role of CD28 signal transduction during APC-dependent T cell activation, we have used Staphylococcal enterotoxins (SEs) presented by a B7-positive APC. We used anti-B7 monoclonal antibodies and a mutant interleukin 2 (IL-2) promoter construct, unresponsive to CD28-generated signals, in transient transfection assays to examine the contribution of the CD28-B7 interaction to IL-2 gene activation. These studies indicate that the CD28-regulated signal transduction pathway is activated during SE stimulation of T cells and plays an important role in SE induction of IL-2 gene expression through its influence upon the CD28-responsive element contained within the IL-2 gene promoter. This effect is particularly profound in the activation of the IL-2 gene in peripheral blood T cells.
Assuntos
Antígenos de Bactérias/imunologia , Antígenos CD/fisiologia , Antígenos de Diferenciação de Linfócitos T/fisiologia , Enterotoxinas/imunologia , Interleucina-2/genética , Receptores de Antígenos de Linfócitos T/fisiologia , Células Apresentadoras de Antígenos/imunologia , Antígenos CD28 , Indução Enzimática , Expressão Gênica , Humanos , Técnicas In Vitro , Proteínas Nucleares/metabolismo , Regiões Promotoras Genéticas , RNA Mensageiro/genética , Transdução de Sinais , Células Tumorais Cultivadas , Fosfolipases Tipo C/metabolismoRESUMO
The TCR is a complex receptor composed of seven polypeptide chains consisting of a ligand-binding subunit, Ti, and a putative signal-transducing subunit, CD3. Phylogenetically conserved charged amino acid residues within the membrane-spanning domains present in all seven chains of the TCR have been proposed to be important in the association between Ti and CD3. Using a Ti beta chain-deficient mutant of the cell line Jurkat, site-directed mutagenesis and transfection of Ti beta chain cDNA was performed to assess the importance of the lysine residue at position 290 within the membrane-spanning domain of the Ti beta chain to expression of the TCR complex. These studies demonstrated that the lysine residue, and not simply conservation of either basic charge or secondary structure, is important at this position.
Assuntos
Lisina , Receptores de Antígenos de Linfócitos T/metabolismo , Northern Blotting , Linhagem Celular , Membrana Celular/metabolismo , Citometria de Fluxo , Humanos , Mutação , Receptores de Antígenos de Linfócitos T alfa-beta , TransfecçãoRESUMO
The defect in diamond formed by a vacancy surrounded by three nearest-neighbor nitrogen atoms and one carbon atom, [Formula: see text], is found in the vast majority of natural diamonds. Despite [Formula: see text] being the earliest electron paramagnetic resonance spectrum observed in diamond, to date no satisfactory simulation of the spectrum for an arbitrary magnetic field direction has been produced due to its complexity. In this work, [Formula: see text] is identified in [Formula: see text]-doped synthetic diamond following irradiation and annealing. The [Formula: see text] spin Hamiltonian parameters are directly determined and used to refine the parameters for [Formula: see text], enabling the latter to be accurately simulated and fitted for an arbitrary magnetic field direction. Study of [Formula: see text] under excitation with green light indicates charge transfer between [Formula: see text] and [Formula: see text]. It is argued that this charge transfer is facilitated by direct ionization of [Formula: see text], an as-yet unobserved charge state of [Formula: see text].
RESUMO
The effects of γ-radiation sterilization on the parenteral excipient l-histidine were analysed by means of EPR spectroscopy. The irradiation process was found to induce the formation of a deamination radical which was persistent in the solid state. The nature and reactivity of the radicals following dissolution in water was evaluated using spin-trapping EPR experiments. The deamination radical was found to regenerate in solution in the presence of trace metals, potentially leading to radical induced degradation reactions occurring up to an hour after the dissolution process. Understanding this process is significant for the improved design of parental pharmaceutical formulations in which unwanted radical reactions after γ-radiation sterilization could lead to degradation of active ingredients.
Assuntos
Excipientes/efeitos da radiação , Radicais Livres/química , Raios gama , Histidina/efeitos da radiação , Esterilização/métodos , Espectroscopia de Ressonância de Spin Eletrônica , Excipientes/química , Histidina/química , PósRESUMO
Defects causing colour in nitrogen-doped chemical vapour-deposited (CVD) diamond can adversely affect the exceptional optical, electronic and spintronic properties of the material. Several techniques were used to study these defects, namely optical absorption spectroscopy, thermoluminescence (TL) and electron paramagnetic resonance (EPR). From our studies, the defects causing colour in nitrogen-doped CVD diamond are clearly not the same as those causing similar colour in natural diamonds. The brown colour arises due to a featureless absorption profile that decreases in intensity with increasing wavelength, and a broad feature at 360 nm (3.49 eV) that scales in intensity with it. Another prominent absorption band, centred at 520 nm (2.39 eV), is ascribed to the neutral nitrogen-vacancy-hydrogen defect. The defects responsible for the brown colour possess acceptor states that are 1.5 eV from the valence band (VB) edge. The brown colour is removed by heat treatment at 1600 ° C, whereupon new defects possessing shallow (<1 eV) trap states are generated.
Assuntos
Diamante/química , Eletrônica , Hidrogênio/química , Nitrogênio/química , Fenômenos Ópticos , Cor , Cristalização , Espectroscopia de Ressonância de Spin Eletrônica , Espectrofotometria Infravermelho , Temperatura , Dosimetria TermoluminescenteAssuntos
Serviço Hospitalar de Nutrição , Alimentos , Ingestão de Alimentos , Humanos , Pacientes , PsicologiaRESUMO
We report on the effects of thermal treatment and ultraviolet irradiation on the point defect concentrations and optical absorption profiles of single crystal CVD synthetic diamond. All thermal treatments were below 850 K, which is lower than the growth temperature and unlikely to result in any structural change. UV-visible absorption spectroscopy measurements showed that upon thermal treatment (823 K), various broad absorption features diminished: an absorption band at 270 nm (used to deduce neutral single substitutional nitrogen (N(S)(0)) concentrations) and also two broad features centred at approximately 360 and 520 nm. Point defect centre concentrations as a function of temperature were also deduced using electron paramagnetic resonance (EPR) spectroscopy. Above â¼500 K, we observed a decrease in the concentration of N(S)(0) centres and a concomitant increase in the negatively charged nitrogen-vacancy-hydrogen (NVH) complex (NVH(-)) concentration. Both transitions exhibited an activation energy between 0.6 and 1.2 eV, which is lower than that for the N(S)(0) donor (â¼1.7 eV). Finally, it was found that illuminating samples with intense short-wave ultraviolet light recovered the N(S)(0) concentration and also the 270, 360 and 520 nm absorption features. From these results, we postulate a valence band mediated charge transfer process between NVH and single nitrogen centres with an acceptor trap depth for NVH of 0.6-1.2 eV. Because the loss of N(S)(0) concentration is greater than the increase in NVH(-) concentration we also suggest the presence of another unknown acceptor existing at a similar energy to NVH. The extent to which the colour in CVD synthetic diamond is dependent on prior history is discussed.
RESUMO
We report on electron paramagnetic resonance (EPR) studies of nitrogen doped diamond that has been (15)N enriched, electron irradiated and annealed. EPR spectra from two new nitrogen containing [Formula: see text] defects are detected and labelled WAR9 and WAR10. We show that the properties of these defects are consistent with them being the ⟨001⟩-nitrogen split interstitial and the ⟨001⟩-nitrogen split interstitial-⟨001⟩-carbon split interstitial pair, respectively. We also provide an explanation for why these defects have previously eluded discovery.
RESUMO
In interphase cells of Aedes aegypti (L.) (2n = 4+ XX/XY), only the nucleolus responded to selective silver staining. The secondary constriction on chromosome 3 remained unresponsive at all times but the six centromeres were identified throughout mitosis from early prophase as well as those stages of meiosis subsequent to diplotene. The centromeric blocks were not synonymous with the pericentric heterochromatin revealed by C-banding. X chromosomes without an intercalary C-band were newly discovered in Ae. aegypti in the Bangalore strain. Sequential Q- or Hoechst 33258/C-banding in this and the Trinidad-30 strain revealed intercalary heterochromatin diversity within and between strains and also differences between intercalary and pericentric heterochromatin.
Assuntos
Aedes/genética , Heterocromatina/ultraestrutura , Aedes/citologia , Animais , Bandeamento Cromossômico , Feminino , Masculino , Meiose , Mitose , Prata , Coloração e Rotulagem , Testículo/citologiaRESUMO
The T-cell antigen receptor (TCR) regulates two signal transduction pathways: the phosphatidylinositol (PtdIns) and tyrosine kinase pathways. Stimulation of T cells with antigen or anti-TCR monoclonal antibodies induces an increase in inositol phosphates and diacylglycerol, the second messengers responsible for the mobilization of cytoplasmic free calcium and activation of protein kinase C-4. The TCR also activates a tyrosine kinase that is not intrinsic to the TCR. The relationship between these two signal transduction pathways and their contribution to later T-cell responses is unclear. Studies using variants of a murine hybridoma suggested that the PtdIns pathway might not be necessary for or be involved in regulating interleukin-2 (IL-2) production. To address the relationship between later T-cell responses and the early biochemical signals, we investigated the ability of a heterologous receptor with defined signal transduction function to induce T-cell activation. The human muscarinic subtype-1 receptor (HM1), which elicits PtdIns metabolism in neuronal cells through a G protein-coupled mechanism, also functionally activates this pathway when expressed in the T-cell line Jurkat-derived host, J-HM1-2.2 (ref.8). We show here that stimulation of HM1 alone induced IL-2 production and IL-2 receptor alpha chain expression. HM1 does not induce the tyrosine kinase pathway, suggesting that this pathway does not directly influence later T cell-activation responses. Instead, our studies indicate that activation of the PtdIns pathway is probably sufficient to induce later T-cell responses.
Assuntos
Ativação Linfocitária , Fosfatidilinositóis/metabolismo , Receptores de Antígenos de Linfócitos T/fisiologia , Linfócitos T/fisiologia , Animais , Carbacol/farmacologia , Linhagem Celular , Ativação Enzimática , Humanos , Interleucina-2/biossíntese , Camundongos , Proteína Quinase C/metabolismo , Proteínas Tirosina Quinases/metabolismo , Receptores de Interleucina-2/biossíntese , Receptores de Interleucina-2/fisiologia , Receptores Muscarínicos/fisiologia , Transdução de Sinais , Linfócitos T/imunologia , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
We report the identification of the vacancy-hydrogen complex in single crystal diamond synthesized by chemical vapor deposition. The S=1 defect is observed by electron paramagnetic resonance in the negative charge state. The hydrogen atom is bonded to one of the carbon atoms neighboring the vacancy. Unlike the analogous defect in silicon, no symmetry lowering reconstruction occurs between the three remaining carbon dangling orbitals. The very small measured hydrogen hyperfine interaction is explained by dipolar coupling between the hydrogen and the unpaired electron probability density delocalized on the three equivalent carbon neighbors.
RESUMO
An early consequence of stimulation of T cells via their Ag receptor is the activation of protein kinase C (PKC). It has recently been shown that PKC activity resides in a family of homologous proteins. Inasmuch as T cells are phenotypically and functionally heterogeneous, we examined the possibility that this heterogeneity may be reflected in differential expression of message for PKC isoenzyme genes. RNA from six leukemic T cell lines was probed for PKC-alpha, -beta, and -gamma message before and after activation. These studies revealed significant differences among these lines. None expressed mRNA for PKC-gamma. Whereas all cells possessed message for PKC-alpha, there was consistent variability in the level expressed. The greatest heterogeneity was seen with PKC-beta. Two cell lines, HUT 78 and HPB-ALL, did not hybridize with the beta probe under any conditions tested. We subsequently used these PKC-beta negative cells to study the role of this isoenzyme in mediating some of the effects seen with phorbol esters that directly bind to and activate PKC. Our results indicate that PKC-beta, which is expressed in some T cells, is not necessary for PMA-induced CD3 or CD4 internalization, IL-2 production, or acquisition of the p55 chain of the IL-2 receptor.
Assuntos
Regulação da Expressão Gênica , Proteína Quinase C/genética , Linfócitos T/enzimologia , Linhagem Celular , Genes , Humanos , Isoenzimas/genética , Isoenzimas/fisiologia , Leucemia de Células T/enzimologia , Leucemia de Células T/genética , Fenótipo , Proteína Quinase C/fisiologia , RNA Mensageiro/metabolismo , Linfócitos T/classificação , Linfócitos T/imunologia , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
We report the identification of the nitrogen-vacancy-hydrogen complex in a freestanding nitrogen-doped isotopically engineered single crystal diamond synthesized by chemical vapor deposition. The hydrogen atom is located in the vacancy of a nearest-neighbor nitrogen-vacancy defect and appears to be bonded to the nitrogen atom maintaining the trigonal symmetry of the center. The defect is observed by electron paramagnetic resonance in the negative charge state in samples containing a suitable electron donor (e.g., substitutional nitrogen N(0)(S)).