Inhibitor of cytochrome c messenger RNA -protein interaction in stimulated rat skeletal muscle

The purpose of this work was to examine the possible mechanisms for the regulation of cytochrome c gene expression in response to increased contractile activity in rat skeletal muscle. The working hypothesis was that increased contractile activity enhances cytochrome c gene expression through a cis-element. A 110% increase in cytochrome c mRNA concentration was observed in tibialis anterior (TA) muscle after 9 days of chronic stimulation. Similar difference (120%) exists between soleus (SO) muscle of higher contractile activity and white vastus lateralis (WV) muscle of lower contractile activity. These results suggest that the endogenous cytochrome c gene expression is regulated by contractile activity. Cytochrome c-reporter genes were injected into skeletal muscles to identify the cis-element that is responsible for the regulation. Although the data was inconclusive, part of it suggested the importance of the 3$\sp\prime$-untranslated region (3$\sp\prime$-UTR) in mediating the response to increased contractile activity.^ RNA gel mobility shift (GMSA) and ultraviolet (UV) cross-linking assays revealed specific RNA-protein interaction in a 50-nucleotide region of the 3$\sp\prime$-UTR in unstimulated TA muscle. Computer analysis predicted a stem-loop structure of 17 nucleotides, which provides a structural basis for RNA-protein interaction. These 17 nucleotides are 100% conserved among rat, mouse and human cytochrome c genes and their 13 pseudogenes, suggesting a functional role for this region. The RNA-protein interaction was significantly less in highly active SO muscle than in inactive WV muscle and was dramatically decreased in stimulated TA muscle due to a protein inhibitor(s) associated with ribosome. It is possible that cytochrome c mRNAs undergoing translation are subject to a compartmentalized regulatory influence.^ The conclusion from these results is that increases in contractile activity induce or activate a protein inhibitor(s) associated with ribosome in rat skeletal muscle. The inhibitor decreases RNA-protein interaction in the 3$\sp\prime$-UTR of cytochrome c mRNA, which may result in increased mRNA stability and/or translation. ^

Involvement of {\it p53\/} in PUVA-induced skin cancers

A combination of psoralen and ultraviolet-A radiation, commonly referred to as "PUVA," is widely used in the treatment of psoriasis. However, PUVA treatment increases the risk of developing skin cancer in psoriasis patients and induces skin cancer in mice. It is, however unknown whether the increased incidence of skin cancer in PUVA treated psoriasis patients is due to the carcinogenic effects of PUVA therapy or due to an indirect effect such as immunosuppression, which can permit the growth of tumors induced by UVB radiation. In this study, we used the p53 tumor suppressor gene as a molecular marker to determine whether PUVA-induced mouse skin cancers contain unique mutations in p53 that are different from UV-induced mutations, and if so, determine whether skin cancers from PUVA treated patients have PUVA-type or UV-type p53 mutations. Since the DNA lesions induced by PUVA are quite different from those induced by UV, we hypothesize that p53 mutations induced by PUVA may also be different from those induced by UV.^ Analysis of PUVA-induced murine skin cancers for p53 mutations revealed that 14 of 15 (93%) missense mutations detected in these cancers were localized at 5$\sp\prime$-TA/5$\sp\prime$-TAT sites, potential sites of psoralen photoadditions. Mutations at these sequences are exceedingly rare in UV-induced murine skin cancers. In addition, PUVA-induced murine skin cancers did not contain UV signature (C $\to$ T or CC $\to$ TT transitions) mutations in p53. These results suggest that PUVA induces unique mutations in p53 that can be distinguished from those induced by UV.^ Next we determined whether SCCs arising in PUVA treated psoriasis patients have PUVA-type or UV-type p53 mutations. The results indicated that 16 of 25 (64%) missense p53 mutations detected in SCCs from PUVA treated patients were located at 5$\sp\prime$-TG, 5$\sp\prime$-TA and 5$\sp\prime$-TT sites, putative sites of psoralen photobinding. Interestingly, about 32% of p53 mutations detected in SCCs from PUVA treated patients had the UV signature. Taken together these results suggest that both PUVA and UVB play a role in the development of SCCs in psoriasis patients undergoing PUVA therapy. ^

Observations on the propagation of internal tidal waves on the continental slope off Sierra Leone

During R/V Meteor-cruise no. 30 4 moorings with 17 current meters were placed on the continental slope of Sierra Leone at depths between 81 and 1058 meters. The observation period started on March 8, 1973, 16.55 hours GMT and lasted 19 days for moorings M30_068MOOR, M30_069MOOR, M30_070MOOR on the slope and 9 days for M30_067MOOR on the shelf. One current meter recorded at location M30_067MOOR for 22 days. Hydrographic data were collected at 32 stations by means of the "Kieler Multi-Meeressonde". Harmonic analysis is applied to the first 15 days of the time series to determine the M2 and S2 tides. By vertically averaging of the Fourier coefficients the field of motion is separated into its barotropic and its baroclinic component. The expected error generated by white Gaussian noise is estimated. To estimate the influence of the particular vertical distribution of the current meters, the barotropic M2 tide is calculated by ommitting and interchanging time series of different moorings. It is shown that only the data of moorings M30_069MOOR, M30_070MOOR and M30_067MOOR can be used. The results for the barotropic M2 tide agree well with the previous publications of other authors. On the slope at a depth of 1000 m there is a free barotropic wave under the influence of the Coriolis-force propagating along the slope with an amplitude of 3.4 cm S**-1. On the shelf, the maximum current is substantially greater (5.8 cm s**-1) and the direction of propagation is perpendicular to the slope. As for the continental slope a separation into different baroclinic modes using vertical eigenmodes is not reasonable, an interpretation of the total baroclinic wave field is tried by means of the method of characteristis. Assuming the continental slope to generate several linear waves, which superpose, baroclinic tidal ellipses are calculated. The scattering of the direction of the major axes M30_069MOOR is in contrast to M30_070MOOR, where they are bundled within an angle of 60°. This is presumably caused by the different character of the bottom topography in the vicinity of the two moorings. A detailed discussion of M30_069MOOR is renounced since the accuracy of the bathymetric chart is not sufficient to prove any relation between waves and topography. The bundeling of the major axes at M30_070MOOR can be explained by the longslope changes of the slope, which cause an energy transfer from the longslope barotropic component to the downslope baroclinic component. The maximum amplitude is found at a depth of 245 m where it is expected from the characteristics originating at the shelf edge. Because of the dominating barotropic tide high coherence is found between most of the current meters. To show the influence of the baroclinic tidal waves, the effect of the mean current is considered. There are two periods nearly opposite longshore mean current. For 128 hours during each of these periods, starting on March 11, 05.00, and March 21, 08.30, the coherences and energy spectra are calculated. The changes in the slope of the characteristics are found in agreement with the changes of energy and coherence. Because of the short periods of nearly constant mean current, some of the calculated differences of energy and coherence are not statistically significant. For the M2 tide a calculation of the ratios of vertically integrated total baroclinic energy and vertically integrated barotropic kinetic energy is carried out. Taking into account both components (along and perpendicular to the slope) the obtained values are 0.75 and 0.98 at the slope and 0.38 at the shelf. If each component is considered separately, the ratios are 0.39 and 1.16 parallel to the slope and 5.1 and 15.85 for the component perpendicular to it. Taking the energy transfer from the longslope component to the doenslope component into account, a simple model yields an energy-ratio of 2.6. Considering the limited application of the theory to the real conditions, the obtained are in agreement with the values calculated by Sandstroem.

Characterization of the Abydos region through OSIRIS high-resolution images in support of CIVA measurements

Context. On 12 November 2014, the European mission Rosetta delivered the Philae lander on the nucleus of comet 67P /Churyumov-Gerasimenko (67P). After the first touchdown, the lander bounced three times before finally landing at a site named Abydos. Aims. We provide a morphologically detailed analysis of the Abydos landing site to support Philae's measurements and to give context for the interpretation of the images coming from the Comet Infrared and Visible Analyser (CIVA) camera system onboard the lander. Methods. We used images acquired by the OSIRIS Narrow Angle Camera (NAC) on 6 December 2014 to perform the analysis of the Abydos landing site, which provided the geomorphological map, the gravitational slope map, the size-frequency distribution of the boulders. We also computed the albedo and spectral reddening maps. Results. The morphological analysis of the region could suggest that Philae is located on a primordial terrain. The Abydos site is surrounded by two layered and fractured outcrops and presents a 0.02 km(2) talus deposit rich in boulders. The boulder size frequency distribution gives a cumulative power-law index of 4.0 + 0.3/0.4, which is correlated with gravitational events triggered by sublimation and /or thermal fracturing causing regressive erosion. The average value of the albedo is 5.8% at lambda(1) = 480.7 nm and 7.4% at lambda(2) = 649.2 nm, which is similar to the global albedos derived by OSIRIS and CIVA, respectively.

Dynamique des structures. Vulnérabilité aux bâtiments. Construction parasismique

La leçon s'addresse à la comprehension du comportement des bâtiments soumis à l'accéleration séismique, et présente une introduction au comportement dynamique de oscillateurs (un ou plusieurs dégrés de liberté), du comportément hystérétique des structures (selon modes de dissipation) et aux paramètres séismiques relevants à la conception parasismique, notamment aux spectres de réponse et de démande, et sa relation avec la capacité de la structure (courbe de capacité) où on peut identifier les niveaux de dommage -ou les critères de performance- pour des intensités séismique prévues au projet. Elle considère aussi les méthodes de définition et détermination de la vulnérabilité, façe aux séismes, des différentes typologies constructives, avec l'inclusion finale des typologies pour les sistèmes de contreventement et recomandations visées à éviter aux mêmes la concentration de dommage d'origine séismique. Lecture's goal focuses in the understanding of the behaviour of buildings under seismic excitation. It presents an introduction of dynamics (single or multiple degrees of freedom oscillators) and the hysteretic behaviour of ductile structures, introducing the seismic parameters relevant to the structural design, mostly in the context of response and demand spectra and their relations with capacity curves of structures. On the capacity curve obtained in pushover analysis, points representing the design objectives in terms of performance levels can be identified and related with seismic demand. Lecture deals also with methods on vulnerability analysis for building construction typologies and the behaviour (and related recommendations) of seismic resistant structural typologies, having the distribution of dissipative energy and damage in mind.

Non-destructive measurements of the optical properties of apples by means of time-resolved reflectance

Time-resolved reflectance is proposed and effectively used for the nondestructive measurement of the optical properties in apples. The technique is based on the detection of the temporal dispersion of a short laser pulse injected into the probed medium. The time-distribution of re-emitted photons interpreted with a solution of the Diffusion equation yields the mean values of the absorption and reduced scattering coefficients of the medium. The proposed technique proved valuable for the measurement of the absorption and scattering spectra of different varieties of apples. No major variations were observed in the experimental data when the fruit was peeled, proving that the measured optical properties are referred to the pulp. The depth of probed volume was determined to be about 2 cm. Finally, the technique proved capable to follow the change in chlorophyll absorption during storage.

Non-destructive measurements of the optical properties of apples by means of time-resolved reflectance

Time-resolved reflectance is proposed and effectively used for the nondestructive measurement of the optical properties in apples. The technique is based on the detection of the temporal dispersion of a short laser pulse injected into the probed medium. The time-distribution of re-emitted photons interpreted with a solution of the Diffusion equation yields the mean values of the absorption and reduced scattering coefficients of the medium. The proposed technique proved valuable for the measurement of the absorption and scattering spectra of different varieties of apples. No major variations were observed in the experimental data when the fruit was peeled, proving that the measured optical properties are referred to the pulp. The depth of probed volume was determined to be about 2 cm. Finally, the technique proved capable to follow the change in chlorophyll absorption during storage.

Neutron features at the UPM neutronics hall

The neutronics hall of the Nuclear Engineering Department at the Polytechnical University of Madrid has been characterized. The neutron spectra and the ambient dose equivalent produced by an 241AmBe source were measured at various source-to-detector distances on the new bench. Using Monte Carlo methods a detailed model of the neutronics hall was designed, and neutron spectra and the ambient dose equivalent were calculated at the same locations where measurements were carried out. A good agreement between measured and calculated values was found.

The use of the ESS Bilbao installations for laser fusion studies

In this work the use of ESS-Bilbao fast neutron lines for irradiation of materials for nuclear fusion is studied. For the comparison of ESS-Bilbao with an inertial fusion facility a simplified model of HiPER chamber has been used. Several positions for irradiation at ESS-Bilbao have been also compared. The material chosen for the damage analysis is silica due to its importance on IFC optics. In this work a detailed comparison between the two facilities for silica irradiation is given. The comparison covers the neutron fluxes, doses, defect production and PKA spectra. This study is also intended as a methodological approach or guideline for future works on other materials.

Diseño y simulación térmica de HB-LEDs con Autodesk y PCB para ensayos

Este PFC es un trabajo muy práctico, los objetivos fueron impuestos por el tutor, como parte del desarrollo de herramientas (software y hardware) que serán utilizados posteriormente a nivel de docencia e investigación. El PFC tiene dos áreas de trabajo, la principal y primera que se expone es la utilización de una herramienta de simulación térmica para caracterizar dispositivos semiconductores con disipador, la segunda es la expansión de una tarjeta de adquisición de datos con unas PCBs diseñadas, que no estaban disponibles comercialmente. Se ha probado y configurado “Autodesk 2013 Inventor Fusion” y “Autodesk 2013 Simulation and Multiphysics” para simulación térmica de dispositivos de alta potencia. Estas aplicaciones son respectivamente de diseño mecánico y simulación térmica, y la UPM dispone actualmente de licencia. En esta parte del proyecto se realizará un manual de utilización, para que se continúe con esta línea de trabajo en otros PFC. Además se han diseñado mecánicamente y simulado térmicamente diodos LED de alta potencia luminosa (High Brightness Lights Emitting Diodes, HB-LEDs), tanto blancos como del ultravioleta cercano (UVA). Las simulaciones térmicas son de varios tipos de LEDs que actualmente se están empleando y caracterizando térmicamente en Proyectos Fin de Carrera y una Tesis doctoral. En la segunda parte del PFC se diseñan y realizan unas placas de circuito impreso (PCB) cuya función es formar parte de sistemas de instrumentación de adquisición automática de datos basados en LabVIEW. Con esta instrumentación se pueden realizar ensayos de fiabilidad y de otro tipo a dispositivos y sistemas electrónicos. ABSTRACT. The PFC is a very practical work, the objectives were set by the tutor, as part of the development of tools (software and hardware) that will be used later at level of teaching and research. The PFC has two parts, the first one explains the use of a software tool about thermal simulation to characterize devices semiconductors with heatsink, and second one is the expansion of card data acquisition with a PCBs designed, which were not available commercially. It has been tested and configured "Autodesk 2013 Inventor Fusion" and "Autodesk 2013 Simulation Multiphysics” for thermal simulation of high power devices. These applications are respectively of mechanical design and thermal simulation, and the UPM has at present license. In this part of the project a manual of use will be realized, so that it is continued by this line of work in other PFC. Also they have been designed mechanically and simulated thermally LEDs light (High Brightness Lights Emitting Diodes , HB- LEDs) both white and ultraviolet. Thermal simulations are several types of LEDs are now being used in thermally characterizing in Thesis and PhD. In the second part of the PFC there are designed and realized circuit board (PCB) whose function is to be a part of instrumentation systems of automatic acquisition based on LabVIEW data. With this instrumentation can perform reliability testing and other electronic devices and systems.

Isolation of a myoglobin molten globule by selective cobalt(III)-induced unfolding

Reaction of the Schiff-base complex [Co(acetylacetonate-ethylenediimine)(NH3)2]+ with metmyoglobin at pH 6.5 yields a partially folded protein containing six Co(III) complexes. Although half of its α-helical secondary structure is retained, absorption and CD spectra indicate that the tertiary structure in both B-F and AGH domains is disrupted in the partially folded protein. In analogy to proton-induced unfolding, it is likely that the loss of tertiary structure is triggered by metal-ion binding to histidines. Cobalt(III)-induced unfolding of myoglobin is unique in its selectivity (other proteins are unaffected) and in allowing the isolation of the partially folded macromolecule (the protein does not refold or aggregate upon removal of free denaturant).

In vitro reconstitution of the photosystem I light-harvesting complex LHCI-730: Heterodimerization is required for antenna pigment organization

Here we describe the in vitro reconstitution of photosystem I light-harvesting complexes with pigments and proteins (Lhca1 and Lhca4) obtained by overexpression of tomato Lhca genes in Escherichia coli. Using Lhca1 and Lhca4 individually for reconstitution results in monomeric pigment-proteins, whereas a combination thereof yields a dimeric complex. Interactions of the apoproteins is highly specific, as reconstitution of either of the two constituent proteins in combination with a light-harvesting protein of photosystem II does not result in dimerization. The reconstituted Lhca1/4, but not complexes obtained with either Lhca1 or Lhca4 alone, closely resembles the native LHCI-730 dimer from tomato leaves with regard to spectroscopic properties, pigment composition, and stoichiometry. Monomeric complexes of Lhca1 or Lhca4 possess lower pigment/protein ratios, indicating that interactions of the two subunits not only facilitates pigment reorganization but also recruitment of additional pigments. In addition to higher averages of chlorophyll a/b ratios in monomeric complexes than in LHCI-730, comparative fluorescence and CD spectra demonstrate that heterodimerization involves preferential ligation of more chlorophyll b.

Sulfur K-edge x-ray absorption spectroscopy: A spectroscopic tool to examine the redox state of S-containing metabolites in vivo

The sulfur K-edge x-ray absorption spectra for the amino acids cysteine and methionine and their corresponding oxidized forms cystine and methionine sulfoxide are presented. Distinct differences in the shape of the edge and the inflection point energy for cysteine and cystine are observed. For methionine sulfoxide the inflection point energy is 2.8 eV higher compared with methionine. Glutathione, the most abundant thiol in animal cells, also has been investigated. The x-ray absorption near-edge structure spectrum of reduced glutathione resembles that of cysteine, whereas the spectrum of oxidized glutathione resembles that of cystine. The characteristic differences between the thiol and disulfide spectra enable one to determine the redox status (thiol to disulfide ratio) in intact biological systems, such as unbroken cells, where glutathione and cyst(e)ine are the two major sulfur-containing components. The sulfur K-edge spectra for whole human blood, plasma, and erythrocytes are shown. The erythrocyte sulfur K-edge spectrum is similar to that of fully reduced glutathione. Simulation of the plasma spectrum indicated 32% thiol and 68% disulfide sulfur. The whole blood spectrum can be simulated by a combination of 46% disulfide and 54% thiol sulfur.

Hybrid characteristics of oligonucleotides consisting of isonucleoside 2′,5′-anhydro-3′-deoxy-3′-(thymin-1-yl)-d-mannitol with different linkage modes

Oligonucleotides consisting of the isonucleoside repeating unit 2′,5′-anhydro-3′-deoxy-3′-(thymin-1-yl)-d-mannitol (4) were synthesized with the monomeric unit 4 incorporated into oligonucleotides as 1′→4′ linkage 4a (oligomer I) or 6′→4′ linkage 4b (oligomer II). The hybrid properties of the two oligonucleotides I and II with their complementary strands were investigated by thermal denaturation and CD spectra. Oligonucleotide I (4a) formed a stable duplex with d(A)14 with a slightly reduced Tm value of 36.6°C, relative to 38.2°C for the control duplex d(T)14/d(A)14, but oligomer II (4b) failed to hybridize with a DNA complementary single strand. The spectrum of the duplex oligomer I/d(A)14 showed a positive CD band at 217 nm and a negative CD band at 248 nm attributable to a B-like conformation. Molecular modeling showed that in the case of oligomer I the C6′ hydroxy group of each unit could be located in the groove area when hybridized to the DNA single strand, which might contribute additional hydrogen bonding to the stability of duplex formation.

Environmental stress sensitivity of an ascorbic acid-deficient Arabidopsis mutant.

L-ascorbic acid (vitamin C) is a powerful reducing agent found in millimolar concentrations in plants, and is proposed to play an important role in scavenging free radicals in plants and animals. However, surprisingly little is known about the role of this antioxidant in plant environmental stress adaptation or ascorbate biosynthesis. We report the isolation of soz1, a semi-dominant ozone-sensitive mutant that accumulates only 30% of the normal ascorbate concentration. The results of genetic approaches and feeding studies show that the ascorbate concentration affects foliar resistance to the oxidizing gas ozone. Consistent with the proposed role for ascorbate in reactive oxygen species detoxification, lipid peroxides are elevated in soz1, but not in wild type following ozone fumigation. We show that the soz1 mutant is hypersensitive to both sulfur dioxide and ultraviolet B irradiation, thus implicating ascorbate in defense against varied environmental stresses. In addition to defining the first ascorbate deficient mutant in plants, these results indicate that screening for ozone-sensitive mutants is a powerful method for identifying physiologically important antioxidant mechanisms and signal transduction pathways. Analysis of soz1 should lead to more information about the physiological roles and metabolism of ascorbate.