Diacylglycerol kinase ζ limits the polarized recruitment of diacylglycerol-enriched organelles to the immune synapse in T cells.

The antigen-induced formation of an immune synapse (IS) between T cells and antigen-presenting cells results in the rapid generation of the lipid second messenger diacylglycerol (DAG) in T cells. Diacylglycerol kinase ζ (DGKζ) converts DAG into phosphatidic acid (PA). Cytotoxic T lymphocytes (CTLs) from mice deficient in DGKζ have enhanced antiviral and antitumor activities, indicating that the amount of DAG controls the effectiveness of the T cell response.

High-sensitivity measurements of multiple kinase activities in live single cells.

Increasing evidence has shown that population dynamics are qualitatively different from single-cell behaviors. Reporters to probe dynamic, single-cell behaviors are desirable yet relatively scarce. Here, we describe an easy-to-implement and generalizable technology to generate reporters of kinase activity for individual cells.

A polarized Ca2+, diacylglycerol and STIM1 signalling system regulates directed cell migration.

Ca(2+) signals control cell migration by regulating forward movement and cell adhesion. However, it is not well understood how Ca(2+)-regulatory proteins and second messengers are spatially organized in migrating cells. Here we show that receptor tyrosine kinase and phospholipase C signalling are restricted to the front of migrating endothelial leader cells, triggering local Ca(2+) pulses, local depletion of Ca(2+) in the endoplasmic reticulum and local activation of STIM1, supporting pulsatile front retraction and adhesion.

Accelerated discovery via a whole-cell model.

To test the promise of whole-cell modeling to facilitate scientific inquiry, we compared growth rates simulated in a whole-cell model with experimental measurements for all viable single-gene disruption Mycoplasma genitalium strains. Discrepancies between simulations and experiments led to predictions about kinetic parameters of specific enzymes that we subsequently validated. These findings represent, to our knowledge, the first application of whole-cell modeling to accelerate biological discovery.

STIM proteins and the endoplasmic reticulum-plasma membrane junctions.

Eukaryotic organelles can interact with each other through stable junctions where the two membranes are kept in close apposition. The junction that connects the endoplasmic reticulum to the plasma membrane (ER-PM junction) is unique in providing a direct communication link between the ER and the PM. In a recently discovered signaling process, STIM (stromal-interacting molecule) proteins sense a drop in ER Ca(2+) levels and directly activate Orai PM Ca(2+) channels across the junction space.

Cracking CRAC.

CRACR2A is a newly discovered Ca-binding protein that regulates store-operated Ca entry (SOCE). CRACR2A enhances SOCE by promoting the binding of the endoplasmic reticulum Ca sensor STIM to Orai, a Ca channel located in the plasma membrane. As intracellular Ca levels rise, CRACR2A binds Ca and triggers SOCE inactivation by dissociating from the Orai–STIM complex.

Ultrabroadband biphotons generated via chirped quasi-phase-matched optical parametric down-conversion.

We generate ultrabroadband biphotons via the process of spontaneous parametric down-conversion (SPDC) in quasi-phase-matched nonlinear gratings that have a linearly chirped wave vector. By using these ultrabroadband biphotons (300-nm bandwidth), we measure the narrowest Hong-Ou-Mandel dip to date, having a full width at half maximum of 7.1 fs.

Observation of self-focusing of light mediated by cubic nonlinearities in potassium titanyl phosphate.

We report our observations of the transient self-narrowing of light beams mediated by dominant Kerr nonlinearities and two-photon absorption in a bulk potassium titanyl phosphate crystal near conditions for second-harmonic generation. Drastic differences between upconversion and downconversion processes are highlighted.

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.

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.

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.

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.

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.

Digital spiral imaging.

A major application of optics is imaging all types of structural, physical, chemical and biological features of matter. Techniques based on most known properties of light have been developed over the years to remotely acquire information about such features. They include the spin angular momentum, encoded in the polarization, but not yet the orbital angular momentum encoded in its spiral spectrum.

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.

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.

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.

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.

Complex soliton-like pattern generation in Potassium Niobate due to noisy, high intensity, input beams.

We report the generation of complex soliton-like patterns in noncritically-phase-matched potassium niobate which occur in random spatial patterns from shot-to-shot. Up to five spots have been generated at input intensities of 10's GW/cm2, many times the single soliton threshold. The mechanism which leads to the symmetry breaking required for the complex patterns is interpreted to be random noise imprinted on the input light.

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.

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.

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.

Properties of quadratic multi-soliton generation near phase-match in periodically poled potassium titanyl phosphate.

The properties of the multi-quadratic-soliton generation process have been investigated both theoretically and experimentally near and on phase-match in non-critically-phase-matched, periodically poled, potassium titanyl phosphate (PPKTP). It was found that multi-soliton generation occurs primarily due to asymmetry in the input beam and at phase-matching. The number of solitons generated depended on the input intensity in a non-trivial way.

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.