TATA-Box Binding Protein O-GlcNAcylation at T114 Regulates Formation of the B-TFIID Complex and Is Critical for Metabolic Gene Regulation.

In eukaryotes, gene expression is performed by three RNA polymerases that are targeted to promoters by molecular complexes. A unique common factor, the TATA-box binding protein (TBP), is thought to serve as a platform to assemble pre-initiation complexes competent for transcription. Here, we describe a novel molecular mechanism of nutrient regulation of gene transcription by dynamic O-GlcNAcylation of TBP. We show that O-GlcNAcylation at T114 of TBP blocks its interaction with BTAF1, hence the formation of the B-TFIID complex, and its dynamic cycling on and off of DNA. Transcriptomic and metabolomic analyses of TBPT114A CRISPR/Cas9-edited cells showed that loss of O-GlcNAcylation at T114 increases TBP binding to BTAF1 and directly impacts expression of 408 genes. Lack of O-GlcNAcylation at T114 is associated with a striking reprogramming of cellular metabolism induced by a profound modification of the transcriptome, leading to gross alterations in lipid storage.

Oral Iptacopan Monotherapy in Paroxysmal Nocturnal Hemoglobinuria.

BACKGROUND: Persistent hemolytic anemia and a lack of oral treatments are challenges for patients with paroxysmal nocturnal hemoglobinuria who have received anti-C5 therapy or have not received complement inhibitors. Iptacopan, a first-in-class oral factor B inhibitor, has been shown to improve hemoglobin levels in these patients. METHODS: In two phase 3 trials, we assessed iptacopan monotherapy over a 24-week period in patients with hemoglobin levels of less than 10 g per deciliter. In the first, anti-C5-treated patients were randomly assigned to switch to iptacopan or to continue anti-C5 therapy. In the second, single-group trial, patients who had not received complement inhibitors and who had lactate dehydrogenase (LDH) levels more than 1.5 times the upper limit of the normal range received iptacopan. The two primary end points in the first trial were an increase in the hemoglobin level of at least 2 g per deciliter from baseline and a hemoglobin level of at least 12 g per deciliter, each without red-cell transfusion; the primary end point for the second trial was an increase in hemoglobin level of at least 2 g per deciliter from baseline without red-cell transfusion. RESULTS: In the first trial, 51 of the 60 patients who received iptacopan had an increase in the hemoglobin level of at least 2 g per deciliter from baseline, and 42 had a hemoglobin level of at least 12 g per deciliter, each without transfusion; none of the 35 anti-C5-treated patients attained the end-point levels. In the second trial, 31 of 33 patients had an increase in the hemoglobin level of at least 2 g per deciliter from baseline without red-cell transfusion. In the first trial, 59 of the 62 patients who received iptacopan and 14 of the 35 anti-C5-treated patients did not require or receive transfusion; in the second trial, no patients required or received transfusion. Treatment with iptacopan increased hemoglobin levels, reduced fatigue, reduced reticulocyte and bilirubin levels, and resulted in mean LDH levels that were less than 1.5 times the upper limit of the normal range. Headache was the most frequent adverse event with iptacopan. CONCLUSIONS: Iptacopan treatment improved hematologic and clinical outcomes in anti-C5-treated patients with persistent anemia - in whom iptacopan showed superiority to anti-C5 therapy - and in patients who had not received complement inhibitors. (Funded by Novartis; APPLY-PNH ClinicalTrials.gov number, NCT04558918; APPOINT-PNH ClinicalTrials.gov number, NCT04820530.).

Analysis and implementation of volume reflection gratings in photorefractive lithium niobate for edge enhancement.

Diffraction from volume reflection gratings written in bulk photorefractive lithium niobate is modeled for the case of longitudinally varying index modulation depths. Numerical solutions to the Helmholtz equation are found in the spatial frequency domain, leading to transfer functions for the volume reflection grating. These transfer functions are then used to show the spatial frequency filtering effect of the volume reflection grating on input light fields containing 2D spatial information. It is shown, first through simulations and then by experiment, that the 0th order transmitted beam undergoes a 2D edge enhancement.

Transport of intensity and phase: applications to digital holography [Invited].

We first review transport of intensity and phase and show their use as a convenient tool to directly determine the unwrapped phase of an imaged object, either through conventional imaging or using digital holography. For both cases, either the traditional transport of intensity and phase, or with a modification, viz., electrically controllable transport of intensity and phase, can be used. The use of digital holography with transport of intensity for 3D topographic mapping of fingermarks coated with columnar thin films is shown as an illustrative application of this versatile technique.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

This feature issue is a continuation of a tradition to follow the conclusion of the Optica Topical Meeting on Digital Holography and 3D Imaging (DH+3D). It addresses current research topics in digital holography and 3D imaging that are also in line with the topics of Applied Optics and Journal of the Optical Society of America A.

Multiplexed digital volume reflection holograms generated from digital transmission holograms.

Interference from co-propagation of the object and reference beams can be digitally recorded for a digital transmission hologram (DTH). Volume holograms, as in display holography, which have been traditionally recorded in bulk photopolymer or photorefractive materials using a counter-propagating object and writing beams, are read out using multispectral light and offer the advantage of excellent wavelength selectivity. In this work, the reconstruction from a single digital volume reflection hologram (DVRH) and wavelength multiplexed DVRHs derived from respective single and multi-wavelength DTHs is investigated, using coupled wave theory and an angular spectral approach. The dependence of the diffraction efficiency on volume grating thickness, wavelength, and incident angle of the reading beam is studied.

Nonlinear optical properties and carrier recombination lifetime of GaN.

Nonlinear optical properties of a selection of gallium nitride samples have been measured using picosecond and nanosecond duration laser pulses at 532 nm. The values of the two-photon absorption coefficient, free carrier absorption cross section, and free carrier refraction cross section are determined along with the recombination lifetime of photogenerated carriers. The effect of hot isostatic pressing on these properties in samples with linear absorption at the band edge due to defects is explored.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

This feature issue is a continuation of a tradition to follow the conclusion of the Optica Topical Meeting on Digital Holography and 3D Imaging (DH+3D). It addresses current research topics in digital holography and 3D imaging that are also in line with the topics of Applied Optics and Journal of the Optical Society of America A.

Excessive O-GlcNAcylation Causes Heart Failure and Sudden Death.

BACKGROUND: Heart failure is a leading cause of death worldwide and is associated with the rising prevalence of obesity, hypertension, and diabetes. O-GlcNAcylation (the attachment of O-linked ß-N-acetylglucosamine [O-GlcNAc] moieties to cytoplasmic, nuclear, and mitochondrial proteins) is a posttranslational modification of intracellular proteins and serves as a metabolic rheostat for cellular stress. Total levels of O-GlcNAcylation are determined by nutrient and metabolic flux, in addition to the net activity of 2 enzymes: O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Failing myocardium is marked by increased O-GlcNAcylation, but whether excessive O-GlcNAcylation contributes to cardiomyopathy and heart failure is unknown. METHODS: We developed 2 new transgenic mouse models with myocardial overexpression of OGT and OGA to control O-GlcNAcylation independent of pathologic stress. RESULTS: We found that OGT transgenic hearts showed increased O-GlcNAcylation and developed severe dilated cardiomyopathy, ventricular arrhythmias, and premature death. In contrast, OGA transgenic hearts had lower O-GlcNAcylation but identical cardiac function to wild-type littermate controls. OGA transgenic hearts were resistant to pathologic stress induced by pressure overload with attenuated myocardial O-GlcNAcylation levels after stress and decreased pathologic hypertrophy compared with wild-type controls. Interbreeding OGT with OGA transgenic mice rescued cardiomyopathy and premature death, despite persistent elevation of myocardial OGT. Transcriptomic and functional studies revealed disrupted mitochondrial energetics with impairment of complex I activity in hearts from OGT transgenic mice. Complex I activity was rescued by OGA transgenic interbreeding, suggesting an important role for mitochondrial complex I in O-GlcNAc-mediated cardiac pathology. CONCLUSIONS: Our data provide evidence that excessive O-GlcNAcylation causes cardiomyopathy, at least in part, attributable to defective energetics. Enhanced OGA activity is well tolerated and attenuation of O-GlcNAcylation is beneficial against pressure overload-induced pathologic remodeling and heart failure. These findings suggest that attenuation of excessive O-GlcNAcylation may represent a novel therapeutic approach for cardiomyopathy.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

This feature issue is a continuation of a tradition, since 2007, to follow the conclusion of the OSA Topical Meeting on Digital Holography and 3D Imaging (DH+3D). It addresses current research topics in digital holography (DH) and 3D imaging that are also in line with the topics of Applied Optics (AO) and the Journal of the Optical Society of America A (JOSA A).

Non-recursive transport of intensity phase retrieval with the transport of phase.

The transport of intensity equation (TIE) is a non-interferometric phase retrieval method that originates from the imaginary part of the Helmholtz equation and is equivalent to the law of conservation of energy. From the real part of the Helmholtz equation, the transport of phase equation (TPE), which represents the Eikonal equation in the presence of diffraction, can be derived. The amplitude and phase for an arbitrary optical field should satisfy these coupled equations simultaneously during propagation. In this work, the coupling between the TIE and TPE is exploited to improve the phase retrieval solutions from the TIE. Specifically, a non-recursive fast Fourier transform (FFT)-based phase retrieval method using both the TIE and TPE is demonstrated. Based on the FFT-based TIE solution, a correction factor calculated by the TPE is introduced to improve the phase retrieval results.

Use of structured light in 3D reconstruction of transparent objects.

A simple non-interferometric incoherent light ray propagation model is introduced to perform three-dimensional profiling of transparent objects with typical thicknesses of the order of mm to cm by analyzing the distorted captured image behind the object. A two-dimensional cosine fringe is used as the incident reference image, whose periodicity is markedly altered by the shape of the object. By monitoring the local change in the period, the surface profile is simulated and optimized to achieve minimal error with experimental data and thus determine the final morphology. Our proposed method is simple, robust, straightforward, and single-shot, and can be used with coherent or incoherent illumination. Its feasibility for more complex applications is verified experimentally through rigorous error calculation. Moreover, the application of this technique for arbitrary transparent objects is theoretically attainable and promising.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

This feature issue is a continuation of a tradition, since 2007, to follow the conclusion of the OSA Topical Meeting on Digital Holography and 3D Imaging (DH+3D). It addresses current research topics in digital holography (DH) and 3D imaging that are also in line with the topics of Applied Optics (AO) and the Journal of the Optical Society of America A (JOSA A).

T-LAK cell-originated protein kinase (TOPK) enhances androgen receptor splice variant (ARv7) and drives androgen-independent growth in prostate cancer.

Despite impressive advances in the treatment of prostate cancer with various efficacious inhibitors along the androgen/androgen receptor axis, eventual development of incurable metastatic Castration-Resistant Prostate Cancer (mCRPC) is inevitable and remains a major clinical challenge. Constitutively active androgen receptor (AR) spliced variants have emerged as primary means of resistance to anti-androgens and androgen synthesis inhibitors. The alternatively spliced AR variant, ARv7, has attracted significant interest due to its constitutively active status in CRPC that drives androgen-independence. Factors that are involved in regulating ARv7 levels in CRPC are not clearly known. We recently demonstrated that a protein kinase, T-LAK cell-originated protein kinase (TOPK) level correlates with the aggressiveness of prostate cancer and its invasive behavior. In this study, we investigated whether TOPK plays a role in driving androgen-independence in prostate cancer cells. Our data demonstrate that TOPK overexpression in androgen-dependent LNCaP and VCaP induces ARv7 and drives androgen-independent growth. On the other hand, pharmacological inhibition of TOPK in androgen-independent LNCaP95 and 22Rv1 represses AR transactivation, and AR stability. In summary, this study illustrates a direct role of TOPK in regulating ARv7 and driving androgen-independence in prostate cancer cells.

Single-shot digital phase-shifting Moiré patterns for 3D topography.

A simple and robust technique of Moiré topography with single-image capture and incorporating digital filtering along with a four-step digitally implemented phase-shifting method is introduced for three-dimensional (3D) surface mapping. Feature details in the order of tens to hundreds of microns can be achieved using interferometrically generated structured light to illuminate the object surface. Compared to the traditional optical phase-shifting method, a digital phase-shifting method based on Fourier processing is implemented with computer-generated sinusoidal patterns derived from the recorded deformed fringes. This enables a single capture of the image that can be used to reconstruct the 3D topography of the surface. Single-shot imaging is simple to implement experimentally and avoids errors in introducing the correct phase shifts. The feasibility of this technique is verified experimentally, and applications to metallic surfaces are demonstrated.

Performance analysis of phase retrieval using transport of intensity with digital holography [Invited].

The performance of direct and unwrapped phase retrieval, which combines digital holography with the transport of intensity, is examined in detail in this paper. In this technique, digital holography is used to numerically reconstruct the intensities at different planes around the image plane, and phase retrieval is achieved by the transport of intensity. Digital holography with transport of intensity is examined for inline and off-axis geometries. The effect of twin images in the inline case is evaluated. Phase-shifting digital holography with transport of intensity is introduced. The performance of digital holography with transport of intensity is compared with traditional off-axis single- and dual-wavelength techniques, which employ standard phase unwrapping algorithms. Simulations and experiments are performed to determine and compare the accuracy of phase retrieval through a mean-squared-error figure of merit as well as the computational speeds of the various methods.

2 × 2 anisotropic transfer matrix approach for optical propagation in uniaxial transmission filter structures.

Multi-layered metamaterial structures show promise in a wide variety of optical applications such as superlenses, electromagnetic cloaking, tunable filters, sensors, and spatial light modulators. Optical transmission analysis of multilayer metallo-dielectric stacks with overall thickness less than the wavelength of light can be modeled using effective medium theory and the Berreman matrix method. For multilayer anisotropic stacks of arbitrary thickness, a rigorous 4 × 4 transfer matrix embodiment is typically used. In this work, a 2 × 2 anisotropic transfer matrix method is developed to analyze optical propagation through multilayer uniaxial stacks of arbitrary thicknesses. Optical transmission of a multilayer silver-zinc oxide stack deposited on a quartz substrate is modeled with this 2 × 2 anisotropic transfer matrix method and reconciled with experimental observations. Results indicate that this numerical approach is applicable to in situ assessment of the complex refractive indices of constituent metal and dielectric layers. Additionally, the anisotropic 2 × 2 transfer matrix method enables the possibility of modeling the transmission of the same metallo-dielectric structure deposited on an electro-optic, uniaxial substrate. Simulation results predict that adjusting the bias field across the substrate results in an electrically tunable transmission filter.

Digital Holography and 3D Imaging: introduction to the joint feature issue in Applied Optics and Journal of the Optical Society of America A.

The OSA Topical Meeting on Digital Holography and 3D Imaging (DH) was held 20-23 May 2019 in Bordeaux, France. Feature issues based on the DH meeting series have been released by Applied Optics (AO) since 2007. This year, AO and the Journal of the Optical Society of America A (JOSA A) jointly decided to have one such feature issue in each journal. This feature issue includes 46 papers in AO and 9 in JOSA A and covers a large range of topics, reflecting the rapidly expanding techniques and applications of digital holography and 3D imaging. The upcoming DH Conference (DH 2020) will be held from 22 to 26 June in Vancouver, Canada, as part of the OSA Imaging and Applied Optics Congress.

3D object recognition through processing of 2D holograms.

Correlation of two-dimensional digitally recorded holograms is introduced as a novel approach for object recognition without the need for quantitative assessment of the retrieved complex field, based on the fact that a hologram contains the three-dimensional information of the object. Actual objects with different three-dimensional features such as depth and surface roughness are assessed through processing of the correlation of their two-dimensional holograms. Correlation peak values are extracted as a metric to evaluate correlation of three-dimensional objects. The effect of hologram windowing size on correlation of three-dimensional objects is investigated, and improvements in computation time and dynamic range are assessed. Critical figures of merit used for assessment of correlation of images are applied to the correlation of holograms for object recognition.

Digital correlation of computer-generated holograms for 3D face recognition.

Three-dimensional (3D) face recognition has been a crucial task in human biometric verification and identification. A digital correlation method of a computer-generated hologram (CGH) for 3D face recognition is proposed, which encodes 3D data into a 2D hologram for recognition. The 3D face models are preprocessed and compressed to into groups of feature points. The CGH templates corresponding to the 3D feature points are generated by point- and layer-oriented algorithms based on three different numerical algorithms to encode depth values into 2D holograms. A 2D digital correlation is performed between the CGH templates. It is demonstrated that the generated CGHs templates could be effectively classified based on the correlation performance metrics of discrimination ratio, peak-to-correlation plane energy, and peak-to-noise ratio. With the essence of the CGH algorithm being the conversion of 3D data to a 2D hologram, the proposed encoding and decoding method has great advantages in reducing computational efforts and potential applications in 3D face recognition, storage, and display.