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Biomedical subjects

Silvia Carrasco

Publications and source records attributed to Silvia Carrasco.

16 recordsLinked to original sources
PubMed2006

Diacylglycerol, when simplicity becomes complex.

Diacylglycerol (DAG) has unique functions as a basic component of membranes, an intermediate in lipid metabolism and a key element in lipid-mediated signaling. In eukaryotes, for example, impaired DAG generation and/or consumption have severe effects on organ development and cell growth associated with diseases such as cancer, diabetes, immune system disorders and Alzheimer's disease. Although DAG has been studied intensively as a signaling lipid, early models of its function are no longer adequate to explain its numerous roles. The interplay between enzymes that control DAG levels, the identification of families of DAG-regulated proteins, and the overlap among DAG metabolic and signaling processes are providing new interpretations of DAG function. Recent discoveries are also delineating the complex and strategic role of DAG in regulating biochemical networks.

Animals↗
PubMed2006

Broadly tunable dual-wavelength light source for coherent anti-Stokes Raman scattering microscopy.

The signal and idler beams from a picosecond, synchronously pumped optical parametric oscillator (OPO) provide the two colors necessary for coherent anti-Stokes Raman scattering (CARS) microscopy. The OPO provides a continuously tunable frequency difference between the two beams over a broad range of Raman shifts (100-3700 cm(-1)) by varying the temperature of a single nonlinear crystal. The near-infrared output (900-1300 nm) allows for deep penetration into thick samples and reduced nonlinear photodamage. Applications of this light source to in vivo cell and ex vivo tissue imaging are demonstrated.

Animals↗
PubMed2006

Broadband light generation by noncollinear parametric downconversion.

Broadband light generation is demonstrated by noncollinear spontaneous parametric downconversion with a cw pump laser. By use of a suitable noncollinear phase-matching geometry and a tightly focused pump beam, downconverted signals that feature a bell-shaped spectral distribution with a bandwidth approaching 200 nm are obtained. As an application of the generated broadband light, submicrometer axial resolution in an optical coherence tomography scheme is demonstrated; a free-space resolution down to 0.8 microm was achieved.

Journal Article↗
PubMed2005

Beta2-chimaerin provides a diacylglycerol-dependent mechanism for regulation of adhesion and chemotaxis of T cells.

The small GTPase Rac contributes to regulation of cytoskeletal rearrangement during chemokine-induced lymphocyte adhesion and migration in a multi-step process that is very precisely coordinated. Chimaerins are Rac1-specific GTPase-activating proteins of unknown biological function, which have a canonical diacylglycerol C1-binding domain. Here we demonstrate endogenous expression of beta2-chimaerin in T lymphocytes and study the functional role of this protein in phorbol ester and chemokine (CXCL12)-regulated T-cell responses. We used green fluorescent protein-tagged beta2-chimaerin and phorbol ester stimulation to investigate changes in protein localization in living lymphocytes. Our results demonstrate that active Rac cooperates with C1-dependent phorbol ester binding to induce sustained GFP-beta2-chimaerin localization to the membrane. Subcellular distribution of GFP beta2-chimaerin in living cells showed no major changes following CXCL12 stimulation. Nonetheless Rac1-GTP levels were severely inhibited in GFP-beta2-chimaerin-expressing cells, which displayed reduced CXCL12-induced integrin-dependent adhesion and spreading. This effect was dependent on chimaerin GTPase-activating protein function and required diacylglycerol generation. Whereas beta2-chimaerin overexpression decreased static adhesion, it enhanced CXCL12-dependent migration via receptor-dependent diacylglycerol production. These studies demonstrate that beta2-chimaerin provides a novel, diacylglycerol-dependent mechanism for Rac regulation in T cells and suggest a functional role for this protein in Rac-mediated cytoskeletal remodeling.

Animals↗
PubMed2005

Engineering of multicolor spatial solitons with chirped-period quasi-phase-matching gratings in optical parametric amplification.

We demonstrate the excitation of solitons in a parametric amplifier with enhanced signal content through the use of a chirped-period quasi-phase-matching grating. This technique affords a low soliton threshold at the input end of a parametric amplifier, and the subsequent transformation to a desired soliton that exists at nonzero wave-vector mismatch through the use of a linearly chirped quasi-phase-matching grating. This approach has an advantage over direct excitation of solitons at nonzero wave-vector mismatch in uniform nonlinear materials and holds potential for improving the efficiency and mode quality of high-gain parametric amplifiers.

Journal Article↗
PubMed2004

Diacylglycerol-dependent binding recruits PKCtheta and RasGRP1 C1 domains to specific subcellular localizations in living T lymphocytes.

Diacylglycerol (DAG) signaling relies on the presence of conserved domain 1 (C1) in its target proteins. Phospholipase C-dependent generation of DAG after T cell receptor (TCR) triggering is essential for the correct immune response onset. Accordingly, two C1-containing proteins expressed in T lymphocytes, Ras guanyl nucleotide-releasing protein1 (RasGRP1) and protein kinase C (PKC), were shown to be fundamental for T-cell activation and proliferation. Although containing the same regulatory domain, they are proposed to relocate to distinct subcellular locations in response to TCR triggering. Here we studied intracellular localization of RasGRP1 and PKC C1 domains in living Jurkat T cells. The results demonstrate that, in the absence of significant primary sequence differences, the C1 domains of these proteins show specific localization within the cell and distinct responses to pharmacological stimulation and TCR triggering. These differences help explain the divergent localization and distinct functional roles of the full-length proteins, which contains them. The properties of these DAG-binding modules allow their characterization as functional markers that discriminate between DAG pools. Finally, we show that by binding to different diacylglycerol forms, overexpression of distinct C1 modules can attenuate DAG-dependent signals originating from the plasma or internal membranes. This is shown by analyzing the contribution of these two lipid pools to PLC-dependent Ras activation in response to TCR triggering.

Amino Acid Motifs↗
PubMed2004

Quasi-phase-matching engineering for spatial control of entangled two-photon states.

We show that transverse engineering of quasi-phase-matched geometries can be used to tailor the spatial mode function that describes the quantum state of photon pairs generated in spontaneous parametric down-conversion. We study several geometries and reveal how properly engineered gratings affect, in particular, the spatial correlations embedded in two-photon entangled states.

Journal Article↗
PubMed2004

Observation of self-trapping of light in walk-off-compensating tandems.

We report the first experimental observation, to our knowledge, of the self-trapping of light in walk-off-compensating optical tandems. The experiment was conducted with picosecond light pulses in a ten-plate optically contacted tandem made of potassium titanyl phosphate prepared for phase matching along a special geometry featuring a huge local walk-off. The observation should open the door to the exploration of multicomponent soliton formation in new classes of materials and settings.

Journal Article↗
PubMed2003

Quadratic soliton self-reflection at a quadratically nonlinear interface.

The reflection of bulk quadratic solutions incident onto a quadratically nonlinear interface in periodically poled potassium titanyl phosphate was observed. The interface consisted of the boundary between two quasi-phase-matched regions displaced from each other by a half-period. At high intensities and small angles of incidence the soliton is reflected.

Journal Article↗
PubMed2003

Deflection of quadratic solitons at edge dislocations.

We report the observation of deflection of optical solitons generated in the frequency doubling of light beams containing edgelike topological amplitude and phase dislocations. The angular deflection of the solitons was found to be controllable through the position of the dislocation. The experiments were conducted near phase matching in a bulk potassium titanyl phosphate crystal pumped with picosecond light pulses at 1064 nm.

Journal Article↗
PubMed2003

Regulation of diacylglycerol kinase alpha by phosphoinositide 3-kinase lipid products.

Diacylglycerol kinase alpha (DAGK alpha), like all type I DAGKs, has calcium regulatory motifs that act as negative regulators of enzyme activity and localization. Accordingly, DAGK alpha is activated by phospholipase C-coupled receptors in a calcium-dependent manner. One of the first functions attributed to DAGK alpha in lymphocytes was that of regulating interleukin 2-induced cell cycle entry. Interleukin-2 nonetheless exerts its action in the absence of cytosolic calcium increase. We have studied alternative receptor-derived signals to explain calcium-independent DAGK alpha activation, and show that DAGK alpha is stimulated by Src-like kinase-dependent phosphoinositide 3 kinase (PI3K) activation in lymphocytes. Our results demonstrate that, in vivo, the increase in cellular levels of PI3K products is sufficient to induce DAGK alpha activation, allowing DAGK alpha relocation to the intact lymphocyte plasma membrane. This activation is isoform-specific, because other type I DAGKs are not subject to this type of regulation. These studies are the first to describe a pathway in which, in the absence of receptor-regulated calcium increase, DAGK alpha activation and membrane localization is a direct consequence of PI3K activation.

Animals↗
PubMed2003

Observation of multiple soliton generation mediated by amplification of asymmetries.

We report the experimental observation of the formation of multiple optical quadratic solitons in a process mediated by the amplification of minute asymmetries in the diffraction properties of the input light. Experiments were conducted in phase-matched second-harmonic generation in a bulk crystal of periodically poled potassium titanyl phosphate pumped at 1064 nm. The different mechanisms that influence the process were investigated numerically, and the pulsed nature of the pump light was found to play a key role in the observed light distributions.

Journal Article↗
PubMed2003

Quadratic spatial solitons in periodically poled KTiOPO4.

The evolution of fundamental beams into quadratic spatial solitons was investigated in a periodically poled bulk KTiOPO4 crystal near and at phase matching. Various soliton properties, such as trapping efficiency as a function of phase mismatch, poling-induced walk-off, and the effects of pulse width, beam width, and input beam quality, were measured. Effects attributed to the presence of cubic nonlinearities were also observed.

Journal Article↗
PubMed2002

Dictyostelium discoideum has a single diacylglycerol kinase gene with similarity to mammalian theta isoforms.

Diacylglycerol kinases (DGKs) phosphorylate the neutral lipid diacylglycerol (DG) to produce phosphatidic acid (PA). In mammalian systems DGKs are a complex family of at least nine isoforms that are thought to participate in down-regulation of DG-based signalling pathways and perhaps activation of PA-stimulated signalling events. We report here that the simple protozoan amoeba Dictyostelium discoideum appears to contain a single gene encoding a DGK enzyme. This gene, dgkA, encodes a deduced protein that contains three C1-type cysteine-rich repeats, a DGK catalytic domain most closely related to the theta subtype of mammalian DGKs and a C-terminal segment containing a proline/glutamine-rich region and a large aspargine-repeat region. This gene corresponds to a previously reported myosin II heavy chain kinase designated myosin heavy chain-protein kinase C (MHC-PKC), but our analysis clearly demonstrates that this protein does not, as suggested by earlier data, contain a protein kinase catalytic domain. A FLAG-tagged version of DgkA expressed in Dictyostelium displayed robust DGK activity. Earlier studies indicating that disruption of this locus alters myosin II assembly levels in Dictyostelium raise the intriguing possibility that DG and/or PA metabolism may play a role in controlling myosin II assembly in this system.

Algorithms↗
PubMed2002

Dynamics of diacylglycerol kinase zeta translocation in living T-cells. Study of the structural domain requirements for translocation and activity.

The diacylglycerol kinases (DGK) regulate diacylglycerol-based signals by phosphorylating this key lipid intermediate to phosphatidic acid. Here, we have investigated the spatial and temporal regulation of diacylglycerol kinase zeta (DGK zeta) in living Jurkat T-cells expressing a muscarinic type I receptor. Using real time confocal videomicroscopy, we show the rapid translocation of a green fluorescent protein-tagged enzyme from the cytosol to the plasma membrane following receptor stimulation. The generation of a panel of truncations, deletions, and point mutations of the enzyme allowed us to examine the requirements of the different structural motifs for both activity and receptor-regulated translocation. The data show that DGK zeta has strict requirements for intact zinc fingers and the conserved catalytic domain for full enzymatic activity. Protein kinase C-driven myristoylated alanine-rich C kinase substrate domain phosphorylation and intact zinc fingers are in turn essential for plasma membrane translocation. DGK zeta does not translocate to the membrane following stimulation of the endogenous T-cell receptor, and our data demonstrate that the specificity in terms of receptor response is provided by the regulatory motifs present at the C-terminal domain of the protein. This is the first report that shows in vivo DGK zeta translocation in response to agonist stimulation and establishes the role of the different domains in enzymatic activity and the selectivity of the response to receptors.

Animals↗