Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases.

Resveratrol, a polyphenol in red wine, has been reported as a calorie restriction mimetic with potential antiaging and antidiabetogenic properties. It is widely consumed as a nutritional supplement, but its mechanism of action remains a mystery. Here, we report that the metabolic effects of resveratrol result from competitive inhibition of cAMP-degrading phosphodiesterases, leading to elevated cAMP levels. The resulting activation of Epac1, a cAMP effector protein, increases intracellular Ca(2+) levels and activates the CamKKß-AMPK pathway via phospholipase C and the ryanodine receptor Ca(2+)-release channel. As a consequence, resveratrol increases NAD(+) and the activity of Sirt1. Inhibiting PDE4 with rolipram reproduces all of the metabolic benefits of resveratrol, including prevention of diet-induced obesity and an increase in mitochondrial function, physical stamina, and glucose tolerance in mice. Therefore, administration of PDE4 inhibitors may also protect against and ameliorate the symptoms of metabolic diseases associated with aging.

Wide area quantitative phase microscopy by spatial phase scanning digital holography.

A new technique of digital holographic microscopy is introduced for large area quantitative phase microscopy, dubbed spatial phase scanning digital holography, where the object specimen in an interferometer is scanned across the tilted reference phase field, while acquiring camera frames at regular intervals. Both the large area scan and phase shift acquisition are achieved in one sweep, using a simple optomechanical system. The technique can be useful in diverse applications such as fast scans of blood smear, cell and tissue cultures, and microelectronic surface profiles.

Peptide switch is essential for Sirt1 deacetylase activity.

In mammals, the Sirtuins are composed of seven Sir2 orthologs (Sirt1-7) with a conserved deacetylase core that utilizes NAD(+) as a cofactor. Interestingly, the deacetylase core of Sirt1 by itself has no catalytic activity. We found within the C-terminal domain a 25 aa sequence that is essential for Sirt1 activity (ESA). Our results indicate that the ESA region interacts with and functions as an "on switch" for the deacetylase core. The endogenous Sirt1 inhibitor DBC1, which also binds to the deacetylase core, competes with and inhibits the ESA region from interacting with the deacetylase core. We discovered an ESA mutant peptide that can bind to the deacetylase core and inhibit Sirt1 in trans. By using this mutant peptide, we were able to inhibit Sirt1 activity and to increase the chemosensitivity of androgen-refractory prostate cancer cells. Therefore, the ESA region is a potential target for development of therapies to regulate Sirt1.

Metabolic sensor AMPK directly phosphorylates RAG1 protein and regulates V(D)J recombination.

The ability to sense metabolic stress is critical for successful cellular adaptation. In eukaryotes, the AMP-activated protein kinase (AMPK), a highly conserved serine/threonine kinase, functions as a critical metabolic sensor. AMPK is activated by the rising ADP/ATP and AMP/ATP ratios during conditions of energy depletion and also by increasing intracellular Ca(2+). In response to metabolic stress, AMPK maintains energy homeostasis by phosphorylating and regulating proteins that are involved in many physiological processes including glucose and fatty acid metabolism, transcription, cell growth, mitochondrial biogenesis, and autophagy. Evidence is mounting that AMPK also plays a role in a number of pathways unrelated to energy metabolism. Here, we identify the recombination-activating gene 1 protein (RAG1) as a substrate of AMPK. The RAG1/RAG2 complex is a lymphoid-specific endonuclease that catalyzes specific DNA cleavage during V(D)J recombination, which is required for the assembly of the Ig and T-cell receptor genes of the immune system. AMPK directly phosphorylates RAG1 at serine 528, and the phosphorylation enhances the catalytic activity of the RAG complex, resulting in increased cleavage of oligonucleotide substrates in vitro, or increased recombination of an extrachromosomal substrate in a cellular assay. Our results suggest that V(D)J recombination can be regulated by AMPK activation, providing a potential new link between metabolic stress and development of B and T lymphocytes.

Quantitative phase-contrast confocal microscope.

We present a quantitative phase-contrast confocal microscope (QPCCM) by combining a line-scanning confocal system with digital holography (DH). This combination can merge the merits of these two different imaging modalities. High-contrast intensity images with low coherent noise, and the optical sectioning capability are made available due to the confocality. Phase profiles of the samples become accessible thanks to DH. QPCCM is able to quantitatively measure the phase variations of optical sections of the opaque samples and has the potential to take high-quality intensity and phase images of non-opaque samples such as many biological samples. Because each line scan is recorded by a hologram that may contain the optical aberrations of the system, it opens avenues for a variety of numerical aberration compensation methods and development of full digital adaptive optics confocal system to emulate current hardware-based adaptive optics system for biomedical imaging, especially ophthalmic imaging. Preliminary experiments with a microscope objective of NA 0.65 and 40 × on opaque samples are presented to demonstrate this idea. The measured lateral and axial resolutions of the intensity images from the current system are ~0.64µm and ~2.70µm respectively. The noise level of the phase profile by QPCCM is ~2.4nm which is better than the result by DH.

Full color natural light holographic camera.

Full-color, three-dimensional images of objects under incoherent illumination are obtained by a digital holography technique. Based on self-interference of two beam-split copies of the object's optical field with differential curvatures, the apparatus consists of a beam-splitter, a few mirrors and lenses, a piezo-actuator, and a color camera. No lasers or other special illuminations are used for recording or reconstruction. Color holographic images of daylight-illuminated outdoor scenes and a halogen lamp-illuminated toy figure are obtained. From a recorded hologram, images can be calculated, or numerically focused, at any distances for viewing.

Single-shot self-interference incoherent digital holography using off-axis configuration.

We propose a single-shot incoherent holographic imaging technique that adopts self-interference incoherent digital holography (SIDH) with slight tilt of the plane mirror in the optical configuration. The limited temporal coherence length of the illumination leads the guide-star hologram of the proposed system to have a Gaussian envelope of elliptical ring shape. The observation shows that the reconstruction by cross correlation with the guide-star hologram achieves better quality than the usual propagation methods. Experimentally, we verify that the hologram and 3D reconstruction can be implemented incoherently with the proposed single-shot off-axis SIDH.

Incoherent digital holographic adaptive optics.

An adaptive optical system based on incoherent digital holography is described. Theoretical and experimental studies show that wavefront sensing and compensation can be achieved by numerical processing of digital holograms of incoherent objects and a guide star, thereby dispensing with the hardware components of conventional adaptive optics systems, such as lenslet arrays and deformable mirrors. The incoherent digital holographic adaptive optics (IDHAO) process is seen to be robust and effective under various ranges of parameters, such as aberration type and strength. Furthermore, low and noisy image signals can be extracted by IDHAO to yield high-quality images with good contrast and resolution, both for point-like and continuous extended objects, illuminated with common incoherent light. Potential applications in astronomical and other imaging systems appear plausible.

Digital holography and 3D imaging: introduction to feature issue.

This feature issue of Applied Optics on Digital Holography and 3D Imaging is the sixth of an approximately annual series. Forty-seven papers are presented, covering a wide range of topics in phase-shifting methods, low coherence methods, particle analysis, biomedical imaging, computer-generated holograms, integral imaging, and many others.

Phase aberration correction by correlation in digital holographic adaptive optics.

We present a phase aberration correction method based on the correlation between the complex full-field and guide-star holograms in the context of digital holographic adaptive optics (DHAO). Removal of a global quadratic phase term before the correlation operation plays an important role in the correction. Correlation operation can remove the phase aberration at the entrance pupil plane and automatically refocus the corrected optical field. Except for the assumption that most aberrations lie at or close to the entrance pupil, the presented method does not impose any other constraints on the optical systems. Thus, it greatly enhances the flexibility of the optical design for DHAO systems in vision science and microscopy. Theoretical studies show that the previously proposed Fourier transform DHAO (FTDHAO) is just a special case of this general correction method, where the global quadratic phase term and a defocus term disappear. Hence, this correction method realizes the generalization of FTDHAO into arbitrary DHAO systems. The effectiveness and robustness of this method are demonstrated by simulations and experiments.

Adaptive optics by incoherent digital holography.

Adaptive optics in astronomical and other imaging systems allows compensation of aberrations introduced by random variations of the refractive index in the imaging path. I propose what I believe is a new type of adaptive optics system that dispenses with the hardware lenslet arrays and deformable mirrors of conventional systems. Theoretical and experimental studies show that wavefront sensing and compensation can be achieved by numerical processing of digital holograms of the incoherent object and a guide star. The incoherent digital holographic adaptive optics is seen to be particularly robust and efficient, with envisioned applications in astronomical imaging, as well as fluorescence microscopy and remote sensing.

Noncontact single-pulse optical method to measure interfacial properties in intact systems.

We introduce a noncontact purely optical approach to measuring the localized surface properties of an interface within a system using a single optical pressure pulse and a time-resolved digital holographic quantitative phase-imaging technique to track the propagating nanometric capillary disturbance. We demonstrate the proposed method's ability to measure the surface energy of deionized water, methanol, and chemical monolayers formed by surfactants with good agreement to published values. The development of this technique boasts immediate application to static and dynamic systems and near-future applications for living biological cell membranes.

Fourier transform digital holographic adaptive optics imaging system.

A Fourier transform digital holographic adaptive optics imaging system and its basic principles are proposed. The CCD is put at the exact Fourier transform plane of the pupil of the eye lens. The spherical curvature introduced by the optics except the eye lens itself is eliminated. The CCD is also at image plane of the target. The point-spread function of the system is directly recorded, making it easier to determine the correct guide-star hologram. Also, the light signal will be stronger at the CCD, especially for phase-aberration sensing. Numerical propagation is avoided. The sensor aperture has nothing to do with the resolution and the possibility of using low coherence or incoherent illumination is opened. The system becomes more efficient and flexible. Although it is intended for ophthalmic use, it also shows potential application in microscopy. The robustness and feasibility of this compact system are demonstrated by simulations and experiments using scattering objects.

Promyelocytic leukemia inhibits adipogenesis, and loss of promyelocytic leukemia results in fat accumulation in mice.

The function of the tumor suppressor promyelocytic leukemia (PML) protein is disrupted in promyelocytic leukemia. PML has been reported to function as a negative regulator of mTOR (mammalian target of rapamycin) and nuclear Akt under some conditions. mTOR and Akt pathways regulate a diverse array of pathways, including those that control insulin signaling, energy metabolism, growth, cellular survival, and lifespan. Although the PML-mTOR/Akt link suggests that PML may have metabolic functions in the whole organism, very little is known about the metabolic functions of PML. Here we report that PML(-/-) mice did not show any significant metabolic defects. There was no impairment in the mTOR/Akt or AMPK signaling in white adipose tissue, liver, or muscle. However, despite having normal food intake and activity levels, PML(-/-) mice gained body weight faster and had more fat mass, particularly subcutaneous fat mass, in the diet-induced obesity model. Using in vitro adipogenesis models, we discovered that PML is a suppressor of adipogenesis. PML expression decreased during adipogenesis and was undetectable in fully differentiated adipocytes. Loss of PML increased expression of the adipogenic transcription factors CCAAT/enhancer binding protein-α and peroxisome proliferator-activated receptor-γ. We found that the Sirt1-NCor-SMRT corepressor complex, which represses pparg transcription, does not bind to the pparg promoter efficiently upon PML depletion. On the basis of these findings, we propose that PML is a negative regulator of the adipogenic transcription factors and that, in times of energy excess, PML may limit fat accumulation by suppressing the differentiation of preadipocytes into adipocytes.

Terahertz digital holography using angular spectrum and dual wavelength reconstruction methods.

Terahertz digital off-axis holography is demonstrated using a Mach-Zehnder interferometer with a highly coherent, frequency tunable, continuous wave terahertz source emitting around 0.7 THz and a single, spatially-scanned Schottky diode detector. The reconstruction of amplitude and phase objects is performed digitally using the angular spectrum method in conjunction with Fourier space filtering to reduce noise from the twin image and DC term. Phase unwrapping is achieved using the dual wavelength method, which offers an automated approach to overcome the 2π phase ambiguity. Potential applications for nondestructive test and evaluation of visually opaque dielectric and composite objects are discussed.

Digital holographic adaptive optics for ocular imaging: proof of principle.

An adaptive optics (AO) ocular imaging system is proposed that is based on the principles of digital holography and dispenses with the wavefront sensor and wavefront modulator of conventional AO systems, thus reducing the optomechanical complexity and cost while increasing speed and resolution. Numerical simulations and proof-of-principle experiments are presented that demonstrate the feasibility.

PML-dependent apoptosis after DNA damage is regulated by the checkpoint kinase hCds1/Chk2.

The promyelocytic leukaemia (PML) gene is translocated in most acute promyelocytic leukaemias and encodes a tumour suppressor protein. PML is involved in multiple apoptotic pathways and is thought to be pivotal in gamma irradiation-induced apoptosis. The DNA damage checkpoint kinase hCds1/Chk2 is necessary for p53-dependent apoptosis after gamma irradiation. In addition, gamma irradiation-induced apoptosis also occurs through p53-independent mechanisms, although the molecular mechanism remains largely unknown. Here, we report that hCds1/Chk2 mediates gamma irradiation-induced apoptosis in a p53-independent manner through an ataxia telangiectasia-mutated (ATM)-hCds1/Chk2-PML pathway. Our results provide the first evidence of a functional relationship between PML and a checkpoint kinase in gamma irradiation-induced apoptosis.

Digital computation of the complex linear canonical transform.

An efficient algorithm for the accurate computation of the linear canonical transform with complex transform parameters and with complex output variable is presented. Sampling issues are discussed and the requirements for different cases given. Simulations are provided to validate the results.

Determination of absorption coefficient by digital holographic measurement of optical excitation.

Digital holographic microscopy produces quantitative phase analysis of a specimen with excellent optical precision. In this study, this imaging method has been used to observe and measure induced thermal lensing by optical excitation. Previous studies have derived these phase shifts from intensity profiles for the determination of photothermal properties of very transparent materials. We have measured physical observables and determined the absorption coefficients of methanol and ethanol with improved precision and accuracy over traditional thermal lens spectroscopy methods.

Potent PDE4 inhibitor activates AMPK and Sirt1 to induce mitochondrial biogenesis.

AMP-activated protein kinase (AMPK) is an evolutionarily conserved energy sensor. Activation of AMPK leads to a number of metabolic benefits, including improved mitochondrial function in skeletal muscle and lowering of serum glucose levels in type-2 diabetes models. However, direct activation of AMPK leads to cardiac enlargement, and an alternative strategy that activates AMPK without affecting the heart is needed. Inhibition of phosphodiesterase 4 (PDE4), which is poorly expressed in the human heart, activates AMPK in other tissues. In a screen to identify novel PDE4 inhibitors, we discovered compound CBU91, which is 5-10 fold more potent than rolipram, the best characterized PDE4 inhibitor. CBU91, like rolipram, is able to activate AMPK and Sirt1 and increase mitochondrial function in myotubes. These findings suggest that activation of AMPK in myotubes is a general property of PDE4 inhibition and that PDE4 inhibition may activate AMPK in metabolically relevant tissues without affecting the heart.