Nicotinamide riboside rescues dysregulated glycolysis and fatty acid ß-oxidation in a human hepatic cell model of citrin deficiency.

Citrin deficiency (CD) is an inborn error of metabolism caused by loss-of-function of the mitochondrial aspartate/glutamate transporter, CITRIN, which is involved in both the urea cycle and malate-aspartate shuttle. Patients with CD develop hepatosteatosis and hyperammonemia but there is no effective therapy for CD. Currently, there are no animal models that faithfully recapitulate the human CD phenotype. Accordingly, we generated a CITRIN knockout HepG2 cell line using Clustered Regularly Interspaced Short Palindromic Repeats/Cas 9 genome editing technology to study metabolic and cell signaling defects in CD. CITRIN KO cells showed increased ammonia accumulation, higher cytosolic ratio of reduced versus oxidized form of nicotinamide adenine dinucleotide (NAD) and reduced glycolysis. Surprisingly, these cells showed impaired fatty acid metabolism and mitochondrial activity. CITRIN KO cells also displayed increased cholesterol and bile acid metabolism resembling those observed in CD patients. Remarkably, normalizing cytosolic NADH:NAD+ ratio by nicotinamide riboside increased glycolysis and fatty acid oxidation but had no effect on the hyperammonemia suggesting the urea cycle defect was independent of the aspartate/malate shuttle defect of CD. The correction of glycolysis and fatty acid metabolism defects in CITRIN KO cells by reducing cytoplasmic NADH:NAD+ levels suggests this may be a novel strategy to treat some of the metabolic defects of CD and other mitochondrial diseases.

Controlled experimental generation of perturbed high-order Ince-Gaussian laser modes.

We present an experimental approach for generating perturbed high-order Ince-Gaussian laser modes by transforming the low and moderate-intensity lobes of high-order Ince-Gaussian (IG) modes into high-intensity lobes and vice versa. This perturbation reshuffles optical energy among the different lobes and generates new, to the best of our knowledge, modulated Ince-Gaussian (MIG) modes. Computer-generated holograms displayed over spatial light modulators were used to modulate the IGMs. Compared to IG modes, MIG modes are generated precisely in a sense that enhances the IG modes and provides a maximum number of highly intense lobes in a particular mode. That enables the newly generated MIG modes to be utilized more efficiently than IG modes in applications such as particle manipulation and optical trapping of microparticles, which exploit highly intense lobes.

Creation of pure longitudinal super-oscillatory spot.

We present a method that creates a super-oscillatory focal spot of a tightly focused radially polarized beam using the concept of a phase mask. Using vector diffraction theory, we report a super-oscillatory focal spot that is much smaller than the diffraction limit and the super-oscillation criterion. The proposed mask works as a special polarization filter that enhances the longitudinal component and filters out the transverse component of radial polarization at focus, permitting the creation of a pure longitudinal super-oscillatory focal spot.

Controllable experimental modulation of high-order Laguerre-Gaussian laser modes.

High-order helical and sinusoidal Laguerre-Gaussian (LG) laser modes have uneven energy distribution among their multiple concentric vortex core rings and lobes, respectively. Here, we explore an experimental method to reshuffle the optical energy among their multiple concentric vortex core rings and lobes of high-order LG modes in a controllable manner. We numerically designed a diffractive optical element displayed over a spatial light modulator to rearrange optical energy among multiple concentric vortex core rings. This changes outer low-intensity concentric vortex core rings into high-intensity vortex core rings of high-order helical LG modes at the Fourier plane. The precise generation of a high-order modulated helical LG laser mode has a maximum number of highly intense concentric vortex core rings compared to known standard helical LG modes. Further, this method is extended to high-order sinusoidal LG modes consisting of both low- and high-intensity lobes to realize modulated sinusoidal LG modes with a maximum number of highly intense lobes in a controllable manner. We envisage that the modulated helical and sinusoidal high-order LG modes may surpass standard LG modes in many applications where highly intense rings and lobes are crucial, as in particle manipulation of micro- and nanoparticles, and optical lithography.

Super-oscillatory spots with different inhomogeneous linear polarized states.

We present the formation of super-oscillatory (SO) spots by tightly focusing the inhomogeneous linear polarized beam of different polarization states. At the entrance pupil of the focusing lens, a suitable phase manipulation in the incident beam results in a small super-oscillatory spot. Our numerical study based on the vectorial diffraction theory shows that SO spots of controllable size and various polarization combinations are possible. We also discuss the effect of the different polarization patterns of the incident beam on the size and energy distribution of the generated SO spots, which are potentially valuable for the orientation determination of single molecules and polarization-resolved imaging. This study reveals more influence of polarization states on the different components of the focused beam under the utilization of the proposed method rather than the usual tight focusing conditions.

Vitamin B12 and folate decrease inflammation and fibrosis in NASH by preventing syntaxin 17 homocysteinylation.

BACKGROUND & AIMS: Several recent clinical studies have shown that serum homocysteine (Hcy) levels are positively correlated, while vitamin B12 (B12) and folate levels are negative correlated, with non-alcoholic steatohepatitis (NASH) severity. However, it is not known whether hyperhomocysteinemia (HHcy) plays a pathogenic role in NASH. METHODS: We examined the effects of HHcy on NASH progression, metabolism, and autophagy in dietary and genetic mouse models, patients, and primates. We employed vitamin B12 (B12) and folate (Fol) to reverse NASH features in mice and cell culture. RESULTS: Serum Hcy correlated with hepatic inflammation and fibrosis in NASH. Elevated hepatic Hcy induced and exacerbated NASH. Gene expression of hepatic Hcy-metabolizing enzymes was downregulated in NASH. Surprisingly, we found increased homocysteinylation (Hcy-lation) and ubiquitination of multiple hepatic proteins in NASH including the key autophagosome/lysosome fusion protein, Syntaxin 17 (Stx17). This protein was Hcy-lated and ubiquitinated, and its degradation led to a block in autophagy. Genetic manipulation of Stx17 revealed its critical role in regulating autophagy, inflammation and fibrosis during HHcy. Remarkably, dietary B12/Fol, which promotes enzymatic conversion of Hcy to methionine, decreased HHcy and hepatic Hcy-lated protein levels, restored Stx17 expression and autophagy, stimulated ß -oxidation of fatty acids, and improved hepatic histology in mice with pre-established NASH. CONCLUSIONS: HHcy plays a key role in the pathogenesis of NASH via Stx17 homocysteinylation. B12/folate also may represent a novel first-line therapy for NASH. LAY SUMMARY: The incidence of non-alcoholic steatohepatitis, for which there are no approved pharmacological therapies, is increasing, posing a significant healthcare challenge. Herein, based on studies in mice, primates and humans, we found that dietary supplementation with vitamin B12 and folate could have therapeutic potential for the prevention or treatment of non-alcoholic steatohepatitis.

Experimental realization of modulated Hermite-Gaussian laser modes: a maximum number of highly intense lobes.

Here, we present an experimental method that redistributes the optical energy among the lobes of high-order standard Hermite-Gaussian (SHG) laser modes in a controlled manner. We numerically designed diffractive optical elements, displayed over a spatial light modulator for redistribution of optical energy that converts low and moderate intense lobes into all highly intense lobes and vice versa at the Fourier plane. Such precise generation of modulated HG (MHG) laser modes offers a maximum number of highly intense lobes compared to SHG modes. Hence, we envisage that MHG beams may surpass SHG beams in many applications, such as particle manipulation and optical lithography, where highly intense lobes play a significant role.

A nexus of miR-1271, PAX4 and ALK/RYK influences the cytoskeletal architectures in Alzheimer's Disease and Type 2 Diabetes.

Alzheimer's Disease (AD) and Type 2 Diabetes (T2D) share a common hallmark of insulin resistance. Reportedly, two non-canonical Receptor Tyrosine Kinases (RTKs), ALK and RYK, both targets of the same micro RNA miR-1271, exhibit significant and consistent functional down-regulation in post-mortem AD and T2D tissues. Incidentally, both have Grb2 as a common downstream adapter and NOX4 as a common ROS producing factor. Here we show that Grb2 and NOX4 play critical roles in reducing the severity of both the diseases. The study demonstrates that the abundance of Grb2 in degenerative conditions, in conjunction with NOX4, reverse cytoskeletal degradation by counterbalancing the network of small GTPases. PAX4, a transcription factor for both Grb2 and NOX4, emerges as the key link between the common pathways of AD and T2D. Down-regulation of both ALK and RYK through miR-1271, elevates the PAX4 level by reducing its suppressor ARX via Wnt/ß-Catenin signaling. For the first time, this study brings together RTKs beyond Insulin Receptor (IR) family, transcription factor PAX4 and both AD and T2D pathologies on a common regulatory platform.

Estrogen-Related Receptor Alpha: An Under-Appreciated Potential Target for the Treatment of Metabolic Diseases.

The estrogen-related receptor alpha (ESRRA) is an orphan nuclear receptor (NR) that significantly influences cellular metabolism. ESRRA is predominantly expressed in metabolically-active tissues and regulates the transcription of metabolic genes, including those involved in mitochondrial turnover and autophagy. Although ESRRA activity is well-characterized in several types of cancer, recent reports suggest that it also has an important role in metabolic diseases. This minireview focuses on the regulation of cellular metabolism and function by ESRRA and its potential as a target for the treatment of metabolic disorders.

Ube3a deficiency inhibits amyloid plaque formation in APPswe/PS1δE9 mouse model of Alzheimer's disease.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive decline in memory and cognitive function. Pathological hallmark of AD includes aberrant aggregation of amyloid beta (Aß) peptide, which is produced upon sequential cleavage of amyloid precursor protein (APP) by ß- and γ -secretases. On the contrary, α-secretase cleaves APP within the Aß sequence and thereby prevents Aß generation. Here, we investigated the role of ubiquitin ligase Ube3a (involved in synaptic function and plasticity) in the pathogenesis of AD using APPswe/PS1δE9 transgenic mouse model and first noticed that soluble pool of Ube3a was age-dependently decreased in AD mouse in comparison with wild type controls. To further explore the role of Ube3a in AD patho-mechanism, we generated brain Ube3a-deficient AD mice that exhibited accelerated cognitive and motor deficits compared with AD mice. Interestingly, these Ube3a-deficient AD mice were excessively obese from their age of 12 months and having shorter lifespan. Biochemical analysis revealed that the Ube3a-deficient AD mice had significantly reduced level of Aß generation and amyloid plaque formation in their brain compared with age-matched AD mice and this effect could be due to the increased activity of α-secretase, ADAM10 (a disintegrin and metalloproteinase-10) that shift the proteolysis of APP towards non-amyloidogenic pathway. These findings suggest that aberrant function of Ube3a could influence the progression of AD and restoring normal level of Ube3a might be beneficial for AD.

Topoisomerase 1 inhibitor topotecan delays the disease progression in a mouse model of Huntington's disease.

Huntington's disease (HD) is a dominantly inherited progressive neurodegenerative disorder caused by the accumulation of polyglutamine expanded mutant huntingtin as inclusion bodies primarily in the brain. After the discovery of the HD gene, considerable progress has been made in understanding the disease pathogenesis and multiple drug targets have been identified, even though currently there is no effective therapy. Here, we demonstrate that the treatment of topotecan, a brain-penetrating topoisomerase 1 inhibitor, to HD transgenic mouse considerably improved its motor behavioural abnormalities along with a significant extension of lifespan. Improvement of behavioural deficits are accompanied with the significant rescue of their progressively decreased body weight, brain weight and striatal volume. Interestingly, topotecan treatment also significantly reduced insoluble mutant huntingtin load in the HD mouse brain. Finally, we show that topotecan treatment to HD mouse not only inhibits the expression of transgenic mutant huntingtin, but also at the same time induces the expression of Ube3a, an ubiquitin ligase linked to the clearance of mutant huntingtin. These findings suggest that topotecan could be a potential therapeutic molecule to delay the progression of HD.

Experimental realization of structured super-oscillatory pulses.

We demonstrate experimentally a generic method for the synthesis of optical femtosecond pulses based on Gaussian, Airy and Hermite-Gauss functions, which are transformed to exhibit fringes with tunable width. The width of the fringes is set in some cases to be much narrower than the inverse of the spectral bandwidth. Such pulses might be useful for ultrafast spectroscopy, coherent control and nonlinear optics.

Optical and Electrical Characterization of Stable p-Type ZnO Thin Films Obtained by Bismuth Doping.

We report the growth of stable p-type ZnO thin films obtained by doping bismuth (Bi) in ZnO and deposited over ITO coated glass substrate. The Bi doped ZnO thin films have been deposited by a sol-gel spin coating method using zinc acetate and bismuth nitrate as main precursors. The structural, optical and electrical properties of annealed Bi doped thin films have been studied in detail using X-ray diffraction (XRD), Atomic force microscopy (AFM), ellipsometry, hot probe system and Hall measurement. The presence of strong diffraction peak along (101) obtained from the XRD spectra shows that the high-quality Bi doped ZnO nanostructures grow along (101) orientation. A number of important micro-structural parameters for the thin films such as grain size, lattice parameters, stress and texture coefficient have been calculated, in order to show the effect of Bi incorporation in ZnO thin film. Further, transmittance has been calculated over the range of 350- 800 nm wavelength regions. The optical band gap of Bi doped ZnO films have also been calculated for different concentrations of Bi using the data taken by an ellipsometer. Hot probe characterization method has been used to ascertain the type of semiconductor thin film and it was observed that films doped with the concentration of 10 mol% Bi show p-type nature that was found to be stable over the period of eight months. Further, in order to calculate the resistivity, hole concentration, and mobility of p-type Bi doped ZnO thin film Hall measurement have been performed.

Novel Transcriptional Mechanisms for Regulating Metabolism by Thyroid Hormone.

The thyroid hormone plays a key role in energy and nutrient metabolisms in many tissues and regulates the transcription of key genes in metabolic pathways. It has long been believed that thyroid hormones (THs) exerted their effects primarily by binding to nuclear TH receptors (THRs) that are associated with conserved thyroid hormone response elements (TREs) located on the promoters of target genes. However, recent transcriptome and ChIP-Seq studies have challenged this conventional view as discordance was observed between TH-responsive genes and THR binding to DNA. While THR association with other transcription factors bound to DNA, TH activation of THRs to mediate effects that do not involve DNA-binding, or TH binding to proteins other than THRs have been invoked as potential mechanisms to explain this discrepancy, it appears that additional novel mechanisms may enable TH to regulate the mRNA expression. These include activation of transcription factors by SIRT1 via metabolic actions by TH, the post-translational modification of THR, the THR co-regulation of transcription with other nuclear receptors and transcription factors, and the microRNA (miR) control of RNA transcript expression to encode proteins involved in the cellular metabolism. Together, these novel mechanisms enlarge and diversify the panoply of metabolic genes that can be regulated by TH.

Rescue of altered HDAC activity recovers behavioural abnormalities in a mouse model of Angelman syndrome.

Angelman syndrome (AS) is a neurodevelopmental disorder characterized by severe intellectual and developmental disabilities. The disease is caused by the loss of function of maternally inherited UBE3A, a gene that exhibits paternal-specific imprinting in neuronal tissues. Ube3a-maternal deficient mice (AS mice) display many classical features of AS, although, the underlying mechanism of these behavioural deficits is poorly understood. Here we report that the absence of Ube3a in AS mice brain caused aberrant increase in HDAC1/2 along with decreased acetylation of histone H3/H4. Partial knockdown of Ube3a in cultured neuronal cells also lead to significant up-regulation of HDAC1/2 and consequent down-regulation of histones H3/H4 acetylation. Treatment of HDAC inhibitor, sodium valproate, to AS mice showed significant improvement in social, cognitive and motor impairment along with restoration of various proteins linked with synaptic function and plasticity. Interestingly, HDAC inhibitor also significantly increased the expression of Ube3a in cultured neuronal cells and in the brain of wild type mice but not in AS mice. These results indicate that anomalous HDAC1/2 activity might be linked with synaptic dysfunction and behavioural deficits in AS mice and suggests that HDAC inhibitors could be potential therapeutic molecule for the treatment of the disease.

Deficiency of Ube3a in Huntington's disease mice brain increases aggregate load and accelerates disease pathology.

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by abnormal expansion of CAG repeats in the gene encoding huntingtin. Mutant huntingtin undergoes proteolytic processing and its N-terminal fragment containing polyglutamine repeat accumulates as inclusion not only in nucleus but also in cytoplasm and neuronal processes. Here, we demonstrate that removal of ubiquitin ligase Ube3a selectively from HD mice brain resulted in accelerated disease phenotype and shorter lifespan in comparison with HD mice. The deficiency of Ube3a in HD mice brain also caused significant increase in global aggregates load, and these aggregates were less ubiquitinated when compared with age-matched HD mice. These Ube3a-maternal deficient HD mice also showed drastic reduction of DARPP-32, a dopamine-regulated phoshphoprotein in their striatum. These results emphasize the crucial role of Ube3a in the progression of HD and its immense potential as therapeutic target.

An integrative approach identified genes associated with drug response in gastric cancer.

Gastric cancer (GC) is the second leading cause of global cancer mortality worldwide. However, the molecular mechanism underlying its carcinogenesis and drug resistance is not well understood. To identify novel functionally important genes that were differentially expressed due to combinations of genetic and epigenetic changes, we analyzed datasets containing genome-wide mRNA expression, DNA copy number alterations and DNA methylation status from 154 primary GC samples and 47 matched non-neoplastic mucosa tissues from Asian patients. We used concepts of 'within' and 'between' statistical analysis to compare the difference between tumors and controls within each platform, and assessed the correlations between platforms. This 'multi-regulated gene (MRG)' analysis identified 126 differentially expressed genes that underwent a combination of copy number and DNA methylation changes. Most genes were located at genomic loci associated with GC. Statistical enrichment analysis showed that MRGs were enriched for cancer, GC and drug response. We analysed several MRGs that previously had not been associated with GC. Knockdown of DDX27, TH1L or IDH3G sensitized cells to epirubicin or cisplatin, and knockdown of RAI14 reduced cell proliferation. Further studies showed that overexpression of DDX27 reduced epirubicin-induced DNA damage and apoptosis. Levels of DDX27 mRNA and protein were increased in early-stage gastric tumors, and may be a potential diagnostic and prognostic marker for GC. In summary, we used an integrative bioinformatics strategy to identify novel genes that are altered in GC and regulate resistance of GC cells to drugs in vitro.

Measurement of acceleration and orbital angular momentum of Airy beam and Airy-vortex beam by astigmatic transformation.

Special beams, including the Airy beam and the vortex-embedded Airy beam, draw much attention due to their unique features and promising applications. Therefore, it is necessary to devise a straightforward method for measuring these peculiar features of the beams with ease. Hence we present the astigmatic transformation of Airy and Airy-vortex beam. The "acceleration" coefficient of the Airy beam is directly determined from a single image by fitting the astigmatically transformed beam to an analytic expression. In addition, the orbital angular momentum of optical vortex in Airy-vortex beam is measured directly using a single image.

Super-Airy beam: self-accelerating beam with intensified main lobe.

We introduce, theoretically and experimentally, the concept of a diffraction-free "super-Airy" beam, in which the main lobe is reduced to nearly half in size with increased intensity in comparison to the main lobe of the optical Airy beam, while maintaining the same transverse acceleration. It is also observed that when the super-Airy main lobe is blocked during propagation, it recovers to the original size faster than the Airy main lobe.