Determining hot-carrier transport dynamics from terahertz emission.

Understanding the ultrafast excitation and transport dynamics of plasmon-driven hot carriers is critical to the development of optoelectronics, photochemistry, and solar-energy harvesting. However, the ultrashort time and length scales associated with the behavior of these highly out-of-equilibrium carriers have impaired experimental verification of ab initio quantum theories. Here, we present an approach to studying plasmonic hot-carrier dynamics that analyzes the temporal waveform of coherent terahertz bursts radiated by photo-ejected hot carriers from designer nano-antennas with a broken symmetry. For ballistic carriers ejected from gold antennas, we find an ~11-femtosecond timescale composed of the plasmon lifetime and ballistic transport time. Polarization- and phase-sensitive detection of terahertz fields further grant direct access to their ballistic transport trajectory. Our approach opens explorations of ultrafast carrier dynamics in optically excited nanostructures.

Quantitative phase contrast imaging with a nonlocal angle-selective metasurface.

Phase contrast microscopy has played a central role in the development of modern biology, geology, and nanotechnology. It can visualize the structure of translucent objects that remains hidden in regular optical microscopes. The optical layout of a phase contrast microscope is based on a 4 f image processing setup and has essentially remained unchanged since its invention by Zernike in the early 1930s. Here, we propose a conceptually new approach to phase contrast imaging that harnesses the non-local optical response of a guided-mode-resonator metasurface. We highlight its benefits and demonstrate the imaging of various phase objects, including biological cells, polymeric nanostructures, and transparent metasurfaces. Our results showcase that the addition of this non-local metasurface to a conventional microscope enables quantitative phase contrast imaging with a 0.02π phase accuracy. At a high level, this work adds to the growing body of research aimed at the use of metasurfaces for analog optical computing.

The expression of dominant negative TCF7L2 in pancreatic beta cells during the embryonic stage causes impaired glucose homeostasis.

OBJECTIVE: Disruption of TCF7L2 in mouse pancreatic ß-cells has generated different outcomes in several investigations. Here we aim to clarify role of ß-cell TCF7L2 and Wnt signaling using a functional-knockdown approach. METHODS: Adenovirus-mediated dominant negative TCF7L2 (TCF7L2DN) expression was conducted in Ins-1 cells. The fusion gene in which TCF7L2DN expression is driven by P TRE3G was utilized to generate the transgenic mouse line TCF7L2DN Tet . The double transgenic line was created by mating TCF7L2DN Tet with Ins2-rtTA, designated as ßTCFDN. ß-cell specific TCF7L2DN expression was induced in ßTCFDN by doxycycline feeding. RESULTS: TCF7L2DN expression in Ins-1 cells reduced GSIS, cell proliferation and expression of a battery of genes including incretin receptors and ß-cell transcription factors. Inducing TCF7L2DN expression in ßTCFDN during adulthood or immediately after weaning generated no or very modest metabolic defect, while its expression during embryonic development by doxycycline feeding in pregnant mothers resulted in significant glucose intolerance associated with altered ß-cell gene expression and reduced ß-cell mass. CONCLUSIONS: Our observations support a cell autonomous role for TCF7L2 in pancreatic ß-cells suggested by most, though not all, investigations. ßTCFDN is a novel model for further exploring the role of TCF7L2 in ß-cell genesis and metabolic homeostasis.

The Wnt signaling pathway effector TCF7L2 controls gut and brain proglucagon gene expression and glucose homeostasis.

The type 2 diabetes risk gene TCF7L2 is the effector of the Wnt signaling pathway. We found previously that in gut endocrine L-cell lines, TCF7L2 controls transcription of the proglucagon gene (gcg), which encodes the incretin hormone glucagon-like peptide-1 (GLP-1). Whereas peripheral GLP-1 stimulates insulin secretion, brain GLP-1 controls energy homeostasis through yet-to-be defined mechanisms. We aim to determine the metabolic effect of a functional knockdown of TCF7L2 by generating transgenic mice that express dominant-negative TCF7L2 (TCF7L2DN) specifically in gcg-expressing cells. The gcg-TCF7L2DN transgenic mice showed reduced gcg expression in their gut and brain, but not in pancreas. Defects in glucose homeostasis were observed in these mice, associated with attenuated plasma insulin levels in response to glucose challenge. The defect in glucose disposal was exacerbated with high-fat diet. Brain Wnt activity and feeding-mediated hypothalamic AMP-activated protein kinase (AMPK) repression in these mice were impaired. Peripheral injection of the cAMP-promoting agent forskolin increased brain ß-cat Ser675 phosphorylation and brain gcg expression and restored feeding-mediated hypothalamic AMPK repression. We conclude that TCF7L2 and Wnt signaling control gut and brain gcg expression and glucose homeostasis and speculate that positive cross-talk between Wnt and GLP-1/cAMP signaling is an underlying mechanism for brain GLP-1 in exerting its metabolic functions.

LRPPRC mutation suppresses cytochrome oxidase activity by altering mitochondrial RNA transcript stability in a mouse model.

LRPPRC (leucine-rich pentatricopeptide repeat-containing) has been shown to be essential for the maturation of COX (cytochrome c oxidase), possibly by stabilizing RNA transcripts of COXI, COXII and COXIII genes encoded in mtDNA (mitochondrial DNA). We established a mouse 'gene-trap' model using ES cells (embryonic stem cells) in which the C-terminus of LRPPRC has been replaced with a ß-geo construct. Mice homozygous for this modification were found to be subject to embryonic lethality, with death before 12.5 dpc (days post-coitum). Biochemical analysis of MEFs (mouse embryonic fibroblasts) isolated from homozygous mutants showed a major decrease in COX activity, with slight reductions in other respiratory chain complexes with mtDNA encoded components. Constructs of LRPPRC containing different numbers of PPRs (pentatricopeptide repeats) were expressed as recombinant proteins and tested for their ability to bind to the COXI mRNA transcript. Full binding required the first 19 PPR motifs. A specific segment of COXI mRNA was identified as the binding target for LRPPRC, encoded by mouse mtDNA nucleotides 5961-6020. These data strongly suggest that LRPPRC is involved in the maturation of COX, and is involved in stabilizing of mitochondrial mRNAs encoding COX transcripts.

Pharmaco-physio-psychologic effect of Ayurvedic oil-dripping treatment using an essential oil from Lavendula angustifolia.

Ayurvedic oil-dripping treatment, Shirodhara, involves the use of medicated herbal sesame oils. In our previous reports, we found that Shirodhara with plain sesame oil induced anxiolysis and an altered state of consciousness (ASC) in healthy subjects. We studied the pharmaco-physio-psychologic effect of Shirodhara with medicated sesame oil including an essential oil from Lavendula angustifolia (lavender) in the present study. Sixteen (16) healthy females (38 +/- 8 years old) were assigned at random to three treatments applied by a robotic oil-dripping system: plain sesame oil (plain Shirodhara), medicated sesame oil with a 0.3 volume % of lavender essential oil (lavender Shirodhara), or the control supine position. Psychophysiologic parameters including the heart rate, skin temperature of the dorsum of hands and feet, as well as anxiety and ASC were monitored, and the rates of change of these items were calculated to assess the psychophysiologic changes brought about by Shirodhara. Lavender Shirodhara showed potent anxiolytic and ASC-inducing or promoting effects, and induced the largest increase in foot skin temperature. The correlation between anxiolysis and ASC, as well as the correlation between these psychologic effects and the elevated foot skin temperature were larger in the lavender Shirodhara than in the other two conditions. It was speculated that the psycho-physiologic effects of lavender Shirodhara would be brought about by three mechanisms: (1) the well-known relaxing action of essential oils from L. angustifolia mediated by olfactory nerves, (2) the pharmacologic action of substances absorbed through the skin or mucosa in the sesame oil or lavender essential oil, and (3) the physiologic effect of sesame oil dripped on the forehead induced by the somato-autonomic reflex through thermosensors or pressure sensors in the skin or hair follicles via the trigeminal cranial nerve. The complicated pharmaco-physio-psychologic action of Ayurvedic oil treatment may provide a useful model for future pharmaco-physio-psychotherapy.

Disruption of a mitochondrial RNA-binding protein gene results in decreased cytochrome b expression and a marked reduction in ubiquinol-cytochrome c reductase activity in mouse heart mitochondria.

Mice homozygous for a defect in the PTCD2 (pentatricopeptide repeat domain protein 2) gene were generated in order to study the role of this protein in mitochondrial RNA metabolism. These mice displayed specific but variable reduction of ubiquinol-cytochrome c reductase complex activity in mitochondria of heart, liver and skeletal muscle due to a decrease in the expression of mitochondrial DNA-encoded cytochrome b, the catalytic core of the complex. This reduction in mitochondrial function has a profound effect on the myocardium, with replacement of ventricular cardiomyocytes by fibro-fatty tissue. Northern blotting showed a reduction in the mRNA for the mitochondrial DNA encoded proteins cytochrome b (cytb) and ND5 (NADH dehydrogenase subunit 5) and an elevation in a combined pre-processed ND5-CYTB transcript. This suggests that the PTCD2 protein is involved in processing RNA transcripts involving cytochrome b derived from mitochondrial DNA. This defines the site for PTCD2 action in mammalian mitochondria and suggests a possible role for dysfunction of this protein in the aetiology of heart failure.

Psychoneuroimmunologic effects of Ayurvedic oil-dripping treatment.

OBJECTIVE: This study assessed the psychoneuroimmunologic changes achieved by Shirodhara, an Ayurvedic treatment, characterized by dripping oil on the forehead, in a randomized, controlled protocol involving a novel approach using a robotic system. METHODS: In the first experiment for the determination of the most appropriate conditions of Shirodhara, 16 healthy females (33 +/- 9 years old) underwent a 30-minute treatment. In the second study, another 16 healthy females (39 +/- 9 years old) were assigned to either the Shirodhara treatment or control supine position for 30 minutes, with monitoring of physiologic, biochemical, immunologic, and psychometric parameters including anxiety and altered states of consciousness (ASC). RESULTS: The subjects receiving Shirodhara treatment showed lowered levels of state anxiety and higher levels of ASC than those in the control position. Plasma noradrenaline and urinary serotonin excretion decreased significantly more after Shirodhara treatment than in the control. Plasma levels of thyrotropin-releasing hormone, dopamine, and natural killer (NK) cell activity were different between control and Shirodhara treatment. The correlation between anxiolysis and the depth of ASC was significant in the Shirodhara treatment group (r = 0.52, p < 0.05, N = 16), while in the control no correlation was obtained (r = 0.13, p = 0.64, N = 16). The increase in foot skin temperature after Shirodhara showed a significant correlation with anxiolysis and the depth of Trance of ASC (r = 0.58, p < 0.01, r = 0.43, p < 0.01, respectively). NK cell activity after Shirodhara treatment showed a significant correlation with anxiolysis and the depth of Trance of ASC (r = 0.33, p < 0.05, r = 0.56, p < 0.01, respectively). CONCLUSIONS: These results indicate that Shirodhara has anxiolytic and ASC-inducing effects, and it promotes a decrease of noradrenaline and exhibits a sympatholytic effect, resulting in the activation of peripheral foot skin circulation and immunopotentiation.

The role of the LRPPRC (leucine-rich pentatricopeptide repeat cassette) gene in cytochrome oxidase assembly: mutation causes lowered levels of COX (cytochrome c oxidase) I and COX III mRNA.

Leigh syndrome French Canadian (LSFC) is a variant of cytochrome oxidase deficiency found in Québec and caused by mutations in the LRPPRC (leucine-rich pentatricopeptide repeat cassette) gene. Northern blots showed that the LRPPRC mRNA levels seen in skeletal muscle>heart>placenta>kidney>liver>lung=brain were proportionally almost opposite in strength to the severity of the enzymic cytochrome oxidase defect. The levels of COX (cytochrome c oxidase) I and COX III mRNA visible on Northern blots were reduced in LSFC patients due to the common (A354V, Ala354-->Val) founder mutation. The amount of LRPPRC protein found in both fibroblast and liver mitochondria from LSFC patients was consistently reduced to <30% of control levels. Import of [(35)S]methionine LRPPRC into rat liver mitochondria was slower for the mutant (A354V) protein. A titre of LRPPRC protein was also found in nuclear fractions that could not be easily accounted for by mitochondrial contamination. [35S]Methionine labelling of mitochondrial translation products showed that the translation of COX I, and perhaps COX III, was specifically reduced in the presence of the mutation. These results suggest that the gene product of LRPPRC, like PET 309p, has a role in the translation or stability of the mRNA for mitochondrially encoded COX subunits. A more diffuse distribution of LRPPRC in LSFC cells compared with controls was evident when viewed by immunofluorescence microscopy, with less LRPPRC present in peripheral mitochondria.

Identification of a gene causing human cytochrome c oxidase deficiency by integrative genomics.

Identifying the genes responsible for human diseases requires combining information about gene position with clues about biological function. The recent availability of whole-genome data sets of RNA and protein expression provides powerful new sources of functional insight. Here we illustrate how such data sets can expedite disease-gene discovery, by using them to identify the gene causing Leigh syndrome, French-Canadian type (LSFC, Online Mendelian Inheritance in Man no. 220111), a human cytochrome c oxidase deficiency that maps to chromosome 2p16-21. Using four public RNA expression data sets, we assigned to all human genes a "score" reflecting their similarity in RNA-expression profiles to known mitochondrial genes. Using a large survey of organellar proteomics, we similarly classified human genes according to the likelihood of their protein product being associated with the mitochondrion. By intersecting this information with the relevant genomic region, we identified a single clear candidate gene, LRPPRC. Resequencing identified two mutations on two independent haplotypes, providing definitive genetic proof that LRPPRC indeed causes LSFC. LRPPRC encodes an mRNA-binding protein likely involved with mtDNA transcript processing, suggesting an additional mechanism of mitochondrial pathophysiology. Similar strategies to integrate diverse genomic information can be applied likewise to other disease pathways and will become increasingly powerful with the growing wealth of diverse, functional genomics data.