Caspase 8: Mediating the effects of a novel proteasome inhibitor, NPI-0052

Targeting the proteasome with the sole FDA approved proteasome inhibitor (PI), bortezomib, has been fruitful in specific cancers. Its success has generated an interest in next-generation PIs that might have a therapeutic advantage in cancers, such as leukemia, where bortezomib monotherapy was less effective. This study focuses on a novel, clinically relevant PI, NPI-0052. Experiments show that NPI-0052 targets chymotrypsin- and caspase-like activities more potently than the trypsin-like activity in leukemia cells. NPI-0052 induced apoptosis, as determined by caspase-3 activation and DNA fragmentation. Using caspase inhibitors and caspase-8 (I9.2) or FADD (I2.1) deficient cells revealed that caspase-8 was essential for NPI-0052-induced apoptosis. NPI-0052 killed cells via a caspase-8-tBid-mitochondrial pathway, relying on caspase-8, whereas bortezomib relies on several caspases. NPI-0052 increased reactive oxygen species (ROS) levels, which contributed towards cytotoxicity since an antioxidant conferred protection. To improve the clinical efficacy of PIs, NPI-0052 was combined with epigenetic anti-cancer agents, histone deacetylase inhibitors (HDACi). NPI-0052 with MS-275 or vorinostat (FDA approved HDACi), synergistically induced apoptosis more effectively than an HDACi/bortezomib regimen in Jurkat cells. Caspase-8 and ROS contributed towards NPI-0052/HDACi cytotoxicity and caspase-8 mediated superoxide production by NPI-0052 or NPI-0052/HDACi. The proximal targets of these agents: proteasome activity and histone acetylation were examined to determine if they contributed towards synergistic effects. HDACi targeted proteasomal β subunits and corresponding catalytic activities responsible for degrading proteins. Immunoblotting showed increases in histone-H3 expression and its acetylation with NPI-0052 or NPI-0052/HDACi in Jurkat and primary cells. Importantly, the hyper-acetylation by NPI-0052 was not detected with bortezomib, suggesting that this effect may be unique to NPI-0052. An antioxidant attenuated histone-H3 expression and acetylation induced by NPI-0052 alone or with HDACi. Furthermore, the hyper-acetylation by NPI-0052 relied on caspase-8. These novel results show that a PI is eliciting classical epigenetic alterations, demonstrated by hyper-acetylation of histone-H3. This alteration was oxidant and caspase-8 dependent. Overall, results reveal that caspase-8 mediates many effects induced by NPI-0052. Data show overlapping activities by NPI-0052 and HDACi which are contributing, along with caspase-8 activation and oxidative stress, to cytotoxic interactions in leukemia cells, reinforcing the potential clinical utility of combining these two compounds. ^

CONFORMATIONAL DYNAMICS OF K-RAS AND H-RAS PROTEINS: IS THERE FUNCTIONAL SPECIFICITY AT THE CATALYTIC DOMAIN?

Ras proteins serve as crucial signaling modulators in cell proliferation through their ability to hydrolyze GTP and exist in a GTP “on” state and GTP “off” state. There are three different human Ras isoforms: H-ras, N-ras and K-ras (4A and 4B). Although their sequence identity is very high at the catalytic domain, these isoforms differ in their ability to activate different effectors and hence different signaling pathways. Much of the previous work on this topic has attributed this difference to the hyper variable region of Ras proteins, which contains most of the sequence variance among the isoforms and encodes specificity for differential distribution in the membrane. However, we hypothesize that sequence variation on lobe II of Ras catalytic domain alters dynamics and leads to differential preference for different effectors or modulators. In this work, we used all atom molecular dynamics to analyze the dynamics in the catalytic domain of H-ras and K-ras. We have also analyzed the dynamics of a transforming mutant of H-ras and K-ras and further studied the dynamics of an effectorselective mutant of H-ras. Collectively we have determined that wild type K-ras is more dynamic than H-ras and that the structure of the effector binding loop more closely resembles that of the T35S Raf-selective mutant, possibly giving us a new view and insight into the v mode of effector specificity. Furthermore we have determined that specific mutations at the same location perturb the conformational equilibrium differently in H-ras and K-ras and that an enhanced oncogenic potential may arise from different structural perturbations for each point mutation of a specific isoform.

Mid-Pliocene palynomorph record of the North Atlantic

The mid-Pliocene was an episode of prolonged global warmth and strong North Atlantic thermohaline circulation, interrupted briefly at circa 3.30 Ma by a global cooling event corresponding to marine isotope stage (MIS) M2. Paleoceanographic changes in the eastern North Atlantic have been reconstructed between circa 3.35 and 3.24 Ma at Deep Sea Drilling Project Site 610 and Integrated Ocean Drilling Program Site 1308. Mg/Ca ratios and d18O from Globigerina bulloides are used to reconstruct the temperature and relative salinity of surface waters, and dinoflagellate cyst assemblages are used to assess variability in the North Atlantic Current (NAC). Our sea surface temperature data indicate warm waters at both sites before and after MIS M2 but a cooling of ~2-3°C during MIS M2. A dinoflagellate cyst assemblage overturn marked by a decline in Operculodinium centrocarpum reflects a southward shift or slowdown of the NAC between circa 3.330 and 3.283 Ma, reducing northward heat transport 23-35 ka before the global ice volume maximum of MIS M2. This will have established conditions that ultimately allowed the Greenland ice sheet to expand, leading to the global cooling event at MIS M2. Comparison with an ice-rafted debris record excludes fresh water input via icebergs in the northeast Atlantic as a cause of NAC decline. The mechanism causing the temporary disruption of the NAC may be related to a brief reopening of the Panamanian Gateway at about this time.

Late Pliocene and Early Pleistocene dinoflagellate cysts and sea surface temperture reconstruction for several IODP and DSDP sites in the North Atlantic

In an attempt to document the palaeoecological affinities of individual extant and extinct dinoflagellate cysts, Late Pliocene and Early Pleistocene dinoflagellate cyst assemblages have been compared with geochemical data from the same samples. Mg/Ca ratios of Globigerina bulloides were measured to estimate the spring-summer sea-surface temperatures from four North Atlantic IODP/DSDP sites. Currently, our Pliocene-Pleistocene database contains 204 dinoflagellate cyst samples calibrated to geochemical data. This palaeo-database is compared with modern North Atlantic and global datasets. The focus lies in the quantitative relationship between Mg/Ca-based (i.e. spring-summer) sea-surface temperature (SSTMg/Ca) and dinoflagellate cyst distributions. In general, extant species are shown to have comparable spring-summer SST ranges in the past and today, demonstrating that our new approach is valid for inferring spring-summer SST ranges for extinct species. For example, Habibacysta tectata represents SSTMg/Ca values between 10° and 15°C when it exceeds 30% of the assemblage, and Invertocysta lacrymosa exceeds 15% when SSTMg/Ca values are between 18.6° and 23.5°C. However, comparing Pliocene and Pleistocene SSTMg/Ca values with present day summer values for the extant Impagidinium pallidum suggests a greater tolerance of higher temperatures in the past. This species occupies more than 5% of the assemblage at SSTMg/Ca values of 11.6-17.9°C in the Pliocene and Pleistocene, whereas present day summer SSTs are around -1.7 to 6.9°C. This observation questions the value of Impagidinium pallidum as reliable indicator of cold waters in older deposits, and may explain its bipolar distribution.

Escape performance of temperate king scallop, Pecten maximus under ocean warming and acidification

Among bivalves, scallops are exceptional due to their capacity to escape from predators by swimming which is provided by rapid and strong claps that are produced by the phasic muscle interspersed with tonic muscle contractions. Based on the concept of oxygen and capacity-limited thermal tolerance, the following hypothesis was tested: ocean warming and acidification (OWA) would induce disturbances in aerobic metabolic scope and extracellular acid-case status and impair swimming performance in temperate scallops. Following long-term incubation under near-future OWA scenarios [20 vs. 10 °C (control) and 0.112 kPa CO2 (hypercapnia) vs. 0.040 kPa CO2 (normocapnic control)], the clapping performance and metabolic rates (MR) were measured in resting (RMR) and fatigued (maximum MR) king scallops, Pecten maximus, from Roscoff, France. Exposure to OA, either alone or combined with warming, left MR and swimming parameters such as the total number of claps and clapping forces virtually unchanged. Only the duration of the escape response was affected by OA which caused earlier exhaustion in hyper- than in normocapnic scallops at 10 °C. While maximum MR was unaffected, warm exposure increased RMR in both normocapnic and hypercapnic P. maximus resulting in similar Q 10 values of ~2.2. The increased costs of maintenance and the observation of strongly reduced haemolymph PO2 levels indicate that at 20 °C scallops have reached the upper thermal pejus range with unbalanced capacities for aerobic energy metabolism. As a consequence, warming to 20 °C decreased mean phasic force during escape performance until fatigue. The observed prolonged recovery time in warm incubated scallops might be a consequence of elevated metabolic costs at reduced oxygen availability in the warmth.

Sediment volume variations inferred from DGPS measurements within Bossons glacier proglacial area (Mont-Blanc massif, France)

This dataset presents Differential Global Positioning System data (DGPS) acquired within the Bossons glacier proglacial area. Bossons glacier is a rapidly retreating glacier and its proglacial area is deglaciated for ~30 years. Bossons stream is one of the outlets of the subglacial drainage system. It starts as a 800 m steep cascade reach, then flows through an area with gentler slope : the Plan des Eaux (PdE). PdE is a 300 m long, 50 m wide proglacial alluvial plain with an increasing channel mobility in the downstream direction but decreasing slope gradient and incision. As it may act a sediment trap, studying periglacial and proglacial erosion processes in the Bossons catchment requires to quantify PdE sediment volume evolution. A several meter-sized block located within Bossons proglacial area was set up as GPS base : its location was measured by one antenna (Topcon Hyper Pro) by performing 600 consecutive measurements throughout one day. A second antenna (Topcon Hyper Pro) was then used to measure XYZ location of points in the proglacial area with a ~2 m grid. Radio communication between the two antennas allowed differential calculations to be automatically carried out on field using the Topcon FC-250 hand controller. This methodology yields 3 cm XY and 1.5 cm Z uncertainties. DGPS data have been acquired through 10 campaigns from 2004 to 2014; campaigns from 2004 to 2008 cover a smaller area than those from 2010 to 2014. Digital Elevation Model (DEM) have been interpolated from DGPS data and difference between two DEMs yields deposited and eroded volume within PdE. Maps of PdE volume variation between two campaigns show that incision mainly occurs in the upper and lower sections where as deposition dominates in the middle section. Deposition, denudation and net rate (deposition rate - denudation rate) are calculated by normalizing volumes by DEM areas. Deposition dominates results with a mean net rate of 29 mm/yr. However, strong inter-annual variability exists and some years are dominated by denudation : -36 mm/yr and -100 mm/yr for 2006 and 2011, respectively. Nonetheless, oldest campaigns (2004 to 2008) were carried out on the lower part part of the alluvial plain and ruling them out to keep only complete DEM (2010 to 2014) yields a mean net rate of ~15 mm/yr. This results is coherent with field observations of both strong deposition (e.g. flood deposits) and strong erosion (e.g. 30 cm incision) evidences. Bossons glacier proglacial area is thus dynamic with year-to-year geormorphological changes but may leans toward increasing its mean elevation through a deposition dominated system.

De la identidad a las intersubjetividades en la comunicación mundo : Elementos para el debate

El documento tiene como pretensión, en un primer momento, establecer líneas de diferenciación entre la ciencia clásica y la ciencia contemporánea, develando las lógicas convencionales de la razón positiva y sus dinámicas instrumentales, que no favorecen los espacios para la comprensión ni el reconocimiento de la misma comprensión (preocupada, ciertamente, por la hiperespecialización y la binarización de la cotidianidad).

Prisiones textuales. Artificio y violencia enla poesía española del barroco

En busca de una nueva lengua artificial, Góngora trata de restituir con medios poéticos una similitud entre palabras y cosas. Así, en el soneto «Prisión del nácar era articulado¼ (1620), en el que artificio y violencia están correlacionados, la artificialidad de la misma descripción corresponde a lo artificial de la belleza descrita. Las técnicas gongorinas desempeñan aquí una función mimológica, por la cual el texto parece hacer visible y audible una violencia que desde siempre subyace en el elogio lírico de la dama. Su imagen «hiperpreciosa¼ debe su belleza sobresaliente a una constricción masculina. Este rasgo es reelaborado por otros poetas barrocos como Quevedo.

Clay mineraology and grain size composition of surface sediments from South Pacific Ocean

The reconstruction of low-latitude ocean-atmosphere interactions is one of the major issues of (paleo-)environmental studies. The trade winds, extending over 20° to 30° of latitude in both hemispheres, between the subtropical highs and the intertropical convergence zone, are major components of the atmospheric circulation and little is known about their long-term variability on geological time-scales, in particular in the Pacific sector. We present the modern spatial pattern of eolian-derived marine sediments in the eastern equatorial and subtropical Pacific (10°N to 25°S) as a reference data set for the interpretation of SE Pacific paleo-dust records. The terrigenous silt and clay fractions of 75 surface sediment samples have been investigated for their grain-size distribution and clay-mineral compositions, respectively, to identify their provenances and transport agents. Dust delivered to the southeast Pacific from the semi- to hyper-arid areas of Peru and Chile is rather fine-grained (4-8 µm) due to low-level transport within the southeast trade winds. Nevertheless, wind is the dominant transport agent and eolian material is the dominant terrigenous component west of the Peru-Chile Trench south of ~ 5°S. Grain-size distributions alone are insufficient to identify the eolian signal in marine sediments due to authigenic particle formation on the sub-oceanic ridges and abundant volcanic glass around the Galapagos Islands. Together with the clay-mineral compositions of the clay fraction, we have identified the dust lobe extending from the coasts of Peru and Chile onto Galapagos Rise as well as across the equator into the doldrums. Illite is a very useful parameter to identify source areas of dust in this smectite-dominated study area.

De la identidad a las intersubjetividades en la comunicación mundo : Elementos para el debate

El documento tiene como pretensión, en un primer momento, establecer líneas de diferenciación entre la ciencia clásica y la ciencia contemporánea, develando las lógicas convencionales de la razón positiva y sus dinámicas instrumentales, que no favorecen los espacios para la comprensión ni el reconocimiento de la misma comprensión (preocupada, ciertamente, por la hiperespecialización y la binarización de la cotidianidad).

Prisiones textuales. Artificio y violencia enla poesía española del barroco

En busca de una nueva lengua artificial, Góngora trata de restituir con medios poéticos una similitud entre palabras y cosas. Así, en el soneto «Prisión del nácar era articulado¼ (1620), en el que artificio y violencia están correlacionados, la artificialidad de la misma descripción corresponde a lo artificial de la belleza descrita. Las técnicas gongorinas desempeñan aquí una función mimológica, por la cual el texto parece hacer visible y audible una violencia que desde siempre subyace en el elogio lírico de la dama. Su imagen «hiperpreciosa¼ debe su belleza sobresaliente a una constricción masculina. Este rasgo es reelaborado por otros poetas barrocos como Quevedo.

De la identidad a las intersubjetividades en la comunicación mundo : Elementos para el debate

El documento tiene como pretensión, en un primer momento, establecer líneas de diferenciación entre la ciencia clásica y la ciencia contemporánea, develando las lógicas convencionales de la razón positiva y sus dinámicas instrumentales, que no favorecen los espacios para la comprensión ni el reconocimiento de la misma comprensión (preocupada, ciertamente, por la hiperespecialización y la binarización de la cotidianidad).

Prisiones textuales. Artificio y violencia enla poesía española del barroco

En busca de una nueva lengua artificial, Góngora trata de restituir con medios poéticos una similitud entre palabras y cosas. Así, en el soneto «Prisión del nácar era articulado¼ (1620), en el que artificio y violencia están correlacionados, la artificialidad de la misma descripción corresponde a lo artificial de la belleza descrita. Las técnicas gongorinas desempeñan aquí una función mimológica, por la cual el texto parece hacer visible y audible una violencia que desde siempre subyace en el elogio lírico de la dama. Su imagen «hiperpreciosa¼ debe su belleza sobresaliente a una constricción masculina. Este rasgo es reelaborado por otros poetas barrocos como Quevedo.

Palynology and dinoflagellate counts of surface sediments, and sea surface conditions, of sites in the North Pacific

Palynological analyses were performed on 53 surface sediment samples from the North Pacific Ocean, including the Bering and Okhotsk Seas (37-64°N, 144°E-148°W), in order to document the relationships between the dinocyst distribution and sea-surface conditions (temperatures, salinities, primary productivity and sea-ice cover). Samples are characterized by concentrations ranging from 18 to 143816 cysts/cm**3 and the occurrence of 32 species. A canonical correspondence analysis (CCA) was carried out to determine the relationship between environmental variables and the distribution of dinocyst taxa. The first and second axes represent, respectively, 47% and 17.8% of the canonical variance. Axis 1 is positively correlated with all parameters except to the sea-ice and primary productivity in August, which are on the negative side. Results indicate that the composition of dinocyst assemblages is mostly controlled by temperature and that all environmental variables are correlated together. The CCA distinguishes 3 groups of dinocysts: the heterotrophic taxa, the genera Impagidinium and Spiniferites as well as the cyst of Pentapharsodinium dalei and Operculodinium centrocarpum. Five assemblage zones can be distinguished: 1) the Okhotsk Sea zone, which is associated to temperate and eutrophic conditions, seasonal upwellings and Amur River discharges. It is characterized by the dominance of O. centrocarpum, Brigantedinium spp. and Islandinium minutum; 2) the Western Subarctic Gyre zone with subpolar and mesotrophic conditions due to the Kamchatka Current and Alaska Stream inflows. Assemblages are dominated by Nematosphaeropsis labyrinthus, Pyxidinopsis reticulata and Brigantedinium spp.; 3) the Bering Sea zone, depicting a subpolar environment, influenced by seasonal upwellings and inputs from the Anadyr and Yukon Rivers. It is characterized by the dominance of I. minutum and Brigantedinium spp.; 4) the Alaska Gyre zone with temperate conditions and nutrient-enriched surface waters, which is dominated by N. labyrinthus and Brigantedinium spp. and 5) the Kuroshio Extension-North Pacific-Subarctic Current zone characterized by a subtropical and oligotrophic environment, which is dominated by O. centrocarpum, N. labyrinthus and warm taxa of the genus Impagidinium. Transfer functions were tested using the modern analog technique (MAT) on the North Pacific Ocean (= 359 sites) and the entire Northern Hemisphere databases ( = 1419 sites). Results confirm that the updated Northern Hemisphere database is suitable for further paleoenvironmental reconstructions, and the best results are obtained for temperatures with an accuracy of +/-1.7 °C.