Search results for "PRECIPITATION"

Determination of enrichment factors for modified RNA in MeRIP experiments

2019

In the growing field of RNA modification, precipitation techniques using antibodies play an important role. However, little is known about their specificities and protocols are missing to assess their effectiveness. Here we present a method to assess enrichment factors after MeRIP-type pulldown experiments, here exemplified with a commercial antibody against N6-methyladenosine (m6A). Testing different pulldown and elution conditions, we measure enrichment factors of 4-5 using m6A-containing mRNAs against an unmodified control of identical sequence. Both types of mRNA carry 32P labels at different nucleotides, allowing their relative quantification in a mixture after digestion to nucleotides…

Recurrent Mutations in the Basic Domain of TWIST2 Cause Ablepharon Macrostomia and Barber-Say Syndromes

2015

Contains fulltext : 153827.pdf (Publisher’s version ) (Open Access) Ablepharon macrostomia syndrome (AMS) and Barber-Say syndrome (BSS) are rare congenital ectodermal dysplasias characterized by similar clinical features. To establish the genetic basis of AMS and BSS, we performed extensive clinical phenotyping, whole exome and candidate gene sequencing, and functional validations. We identified a recurrent de novo mutation in TWIST2 in seven independent AMS-affected families, as well as another recurrent de novo mutation affecting the same amino acid in ten independent BSS-affected families. Moreover, a genotype-phenotype correlation was observed, because the two syndromes differed based s…

Molecular and structural characterization of fluorescent human parvovirus B19 virus-like particles

2005

Although sharing a T = 1 icosahedral symmetry with other members of the Parvoviridae family, it has been suggested that the fivefold channel of the human parvovirus B19 VP2 capsids is closed at its outside end. To investigate the possibility of placing a relatively large protein moiety at this site of B19, fluorescent virus-like particles (fVLPs) of B19 were developed. The enhanced green fluorescent protein (EGFP) was inserted at the N-terminus of the structural protein VP2 and assembly of fVLPs from this fusion protein was obtained. Electron microscopy revealed that these fluorescent protein complexes were very similar in size when compared to wild-type B19 virus. Further, fluorescence cor…

A Short Caspase-3 Isoform Inhibits Chemotherapy-Induced Apoptosis by Blocking Apoptosome Assembly

2011

Alternative splicing of caspase-3 produces a short isoform caspase-3s that antagonizes caspase-3 apoptotic activity. However, the mechanism of apoptosis inhibition by caspase-3s remains unknown. Here we show that exogenous caspase-3 sensitizes MCF-7 and HBL100 breast cancers cells to chemotherapeutic treatments such as etoposide and methotrexate whereas co-transfection with caspase-3s strongly inhibits etoposide and methotrexate-induced apoptosis underlying thus the anti-apoptotic role of caspase-3s. In caspase-3 transfected cells, lamin-A and α-fodrin were cleaved when caspase-3 was activated by etoposide or methotrexate. When caspase-3s was co-transfected, this cleavage was strongly reduc…

Irreversible gelation of thermally unfolded proteins:structural and mechanical properties of lysozyme aggregates

2010

The formation of protein aggregates is important in many fields of life science and technology. The morphological and mechanical properties of protein solutions depend upon the molecular conformation and thermodynamic and environmental conditions. Non-native or unfolded proteins may be kinetically trapped into irreversible aggregates and undergo precipitation or gelation. Here, we study the thermal aggregation of lysozyme in neutral solutions. We characterise the irreversible unfolding of lysozyme by differential scanning calorimetry. The structural properties of aggregates and their mechanisms of formation with the eventual gelation are studied at high temperature by spectroscopic, rheolog…

Calorimetric investigation of the formation of ZnS nanoparticles in w/o microemulsions

1998

The enthalpies of precipitation of ZnS nanoparticles within water containing reversed micelles of sodium bis(2-ethylhexyl) solfosuccinate, L-α phosphatidylcholine, tetraethyleneglycol-mono-n-dodecyl ether and didodecyldimethylammonium bromide as a function of the molar concentration ratioR (R=[water]/[surfactant]) were measured by calorimetric technique. The results indicate that the energetic state of ZnS nanoparticles confined in the aqueous core of the reversed micelles is different from that in bulk water. Effects due to nanoparticle size, adsorption of HS− ions on the nanoparticle surface and interactions between nanoparticles and water/surfactant interfaces are discussed.

Precipitation of slightly soluble silver salts in reversed AOT micelles: Calorimetric investigation

1991

The enthalpies of precipitation, of AgCl, Agl, Ag2S and silver tetraphenylborate (AgTPB) in water containing reversed sodium bis(2-ethylhexyl) sulfosuccinate (AOT) micelles as a function of the molar concentration ratio R (R=[water]/[AOT]) at various concentrations of AOT and reagent salts, were measured by a calorimetric technique. The molar enthalpies of precipitation are independent of the surfactant concentration but are dependent on the concentration of the reagent salts and the R value. The molar enthalpies for the same processes in bulk water are not approached even at the highest R value. Effects due to the smallness of the microcrystals and to the interactions between ions and mice…

Intraseasonal variations of June?September rainfall and upper-air circulation over Kenya

1996

In the Northern Summer, Kenya is located under the influence of the divergent Indian monsoon flow, and therefore is dry except for two separate areas: the coastal strip and the western regions. Analysis of daily rainfall data for June–September 1982 to 1988 has revealed that, although there are many distinct rainfall events between the two regions, an out-of-phase relationship is also evident, rain on the Coast being frequently accompanied by a drop in the precipitation over the Rift Valley area. It is shown that two types of wind forcing accompany these patterns. Alternating westerly and easterly anomalies at the 700 hPa level are associated with persistent wet and dry conditions (respecti…

The Stationarity of Lead-Lag Teleconnections with East Africa Rainfall and its Incidence on Seasonal Predictability

2001

East Africa experiences large interannual rainfall variations, which can lead to severe droughts, as in 1984 in Ethiopia, or extensive flooding, as in 1997 in Somalia and Kenya. In the last 15 years, significant advances have been made in relating these variations to large-scale ocean-atmosphere anomalies, of which those associated to ENSO (El Nino Southern Oscillation) come first. The additional importance of sea-surface variations in the Tropical Atlantic and Indian Oceans is becoming established. Although 20th century East African precipitation series did not experience decadal-scale trends as significant as those found in the sahelian belt, the correlation with ENSO, the Indian monsoon …

2016

AbstractAuthigenic carbonate build-ups develop at seafloor methane-seeps, where microbially mediated sulphate-dependent anaerobic oxidation of methane facilitates carbonate precipitation. Despite being valuable recorders of past methane seepage events, their role as archives of atmospheric processes has not been examined. Here we show that cyclic sedimentation pulses related to the Indian monsoon in concert with authigenic precipitation of methane-derived aragonite gave rise to a well-laminated carbonate build-up within the oxygen minimum zone off Pakistan (northern Arabian Sea). U–Th dating indicates that the build-up grew during past ~1,130 years, creating an exceptional high-resolution a…