Arrested versus active silica diagenesis reaction boundaries—A review of seismic diagnostic criteria

This paper evaluates previously proposed diagnostic criteria that can be used to determine whether or not there is active migration of the opal-A to opal-CT transition zone (TZA/CT). The criteria are based on the interpretation of 2D and 3D seismic surveys and are therefore geometrical. They involve an assessment of the relationship of the TZA/CT with polygonal fault systems, differential compaction structures and tectonic folds. The most robust evidence for an inactive ‘reaction front’ between opal-A and opal-CT bearing sediments is the discordance of the TZA/CT relative to present-day isotherms. Any of these may be persuasive as diagnostic criteria for the upward arrest of the diagenetic transformation at a regional scale, but actual truncation of the TZA/CT at the modern seabed is definitive for arrested diagenesis. This study argues that diagenetic assessment based solely on a single criterion independently is not reliable as an indicator for the current state of a silica transition. As a conclusion, the analysed seismic/structural criteria should be synthesised to provide a more credible interpretation for silica diagenesis. The use of modern 2D and 3D seismic data for the reconstruction of the diagenetic history of opaline silica bearing sediments offers a new approach to the study of silica diagenesis at a regional scale

Physicochemical and Biological Evaluation of siRNA Polyplexes Based on PEGylated Poly(amido amine)s

PURPOSE: Use of RNA interference as novel therapeutic strategy is hampered by inefficient delivery of its mediator, siRNA, to target cells. Cationic polymers have been thoroughly investigated for this purpose but often display unfavorable characteristics for systemic administration, such as interactions with serum and/or toxicity. METHODS: We report the synthesis of a new PEGylated polymer based on biodegradable poly(amido amine)s with disulfide linkages in the backbone. Various amounts of PEGylated polymers were mixed with their unPEGylated counterparts prior to polyplex formation to alter PEG content in the final complex. RESULTS: PEGylation effectively decreased polyplex surface charge, salt- or serum-induced aggregation and interaction with erythrocytes. Increasing amount of PEG in formulation also reduced its stability against heparin displacement, cellular uptake and subsequent silencing efficiency. Yet, for polyplexes with high PEG content, significant gene silencing efficacy was found, which was combined with almost no toxicity. CONCLUSIONS: PEGylated poly(amido amine)s are promising carriers for systemic siRNA delivery in vivo

Dioctadecyldimethylammonium:monoolein nanocarriers for efficient in vitro gene silencing

This study describes a novel liposomal formulation for siRNA delivery, based on the mixture of the neutral lipid monoolein (MO) and cationic lipids of the dioctadecyldimethylammonium (DODA) family. The cationic lipids dioctadecyldimethylammonium bromide (DODAB) and chloride (DODAC) were compared in order to identify which one will most efficiently induce gene silencing. MO has a fluidizing effect on DODAC and DODAB liposomes, although it was more homogeneously distributed in DODAC bilayers. All MO-based liposomal formulations were able to efficiently encapsulate siRNA. Stable lipoplexes of small size (100-160 nm) with a positive surface charge (>+45 mV) were formed. A more uniform MO incorporation in DODAC:MO may explain an increase of the fusogenic potential of these liposomes. The siRNA-lipoplexes were readily internalized by human nonsmall cell lung carcinoma (H1299) cells, in an energy dependent process. DODAB:MO nanocarriers showed a higher internalization efficiency in comparison to DODAC:MO lipoplexes, and were also more efficient in promoting gene silencing. MO had a similar gene silencing ability as the commonly used helper lipid 1,2-dioleyl-3-phosphatidylethanolamine (DOPE), but with much lower cytotoxicity. Taking in consideration all the results presented, DODAB:MO liposomes are the most promising tested formulation for systemic siRNA delivery.This work was supported by FEDER through POFC - COMPETE and by national funds from FCT through the projects PEst-C/BIA/UI4050/2011 (CBM.A), PEst-C/FIS/UI0607/2011 (CFUM), and PTDC/QUI/69795/2006, while Ana Oliveira holds scholarship SFRH/BD/68588/2010. Eloi Feitosa thanks FAPESP (2011/03566-0) and CNPq (303030/2012-7), and Renata D. Adati thanks FAPESP for scholarship (2011/07414-0). K. Raemdonck is a postdoctoral fellow of the Research Foundation - Flanders (FWO-Vlaanderen). We acknowledge NanoDelivery-I&D em Bionanotecnologia, Lda. for access to their equipment

Nanoparticle Orientation to Control RNA Loading and Ligand Display on Extracellular Vesicles for Cancer Regression

Nanotechnology offers many benefits, and here we report an advantage of applying RNA nanotechnology for directional control. The orientation of arrow-shaped RNA was altered to control ligand display on extracellular vesicle membranes for specific cell targeting, or to regulate intracellular trafficking of small interfering RNA (siRNA) or microRNA (miRNA). Placing membrane-anchoring cholesterol at the tail of the arrow results in display of RNA aptamer or folate on the outer surface of the extracellular vesicle. In contrast, placing the cholesterol at the arrowhead results in partial loading of RNA nanoparticles into the extracellular vesicles. Taking advantage of the RNA ligand for specific targeting and extracellular vesicles for efficient membrane fusion, the resulting ligand-displaying extracellular vesicles were capable of specific delivery of siRNA to cells, and efficiently blocked tumour growth in three cancer models. Extracellular vesicles displaying an aptamer that binds to prostate-specific membrane antigen, and loaded with survivin siRNA, inhibited prostate cancer xenograft. The same extracellular vesicle instead displaying epidermal growth-factor receptor aptamer inhibited orthotopic breast cancer models. Likewise, survivin siRNA-loaded and folate-displaying extracellular vesicles inhibited patient-derived colorectal cancer xenograft

Research Data

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Temperature–time relationships and their implications for thermal history and modelling of silica diagenesis in deep-sea sediments

67 Ocean Drilling Program and Deep Sea Drilling Project sites were investigated to determine the relationship between temperature and time for silica diagenesis. The selected sites cover a variety of settings where the opal-A to opal-CT transition zone lies in Cenozoic sediments. The opal-A to opal-CT transition leads to abrupt changes in the sediment petrophysics which are used to identify the diagenetic interval at the study sites. This transition zone is a thin interval ranging in thickness from 10 to 40 m. Controls on transition zone thickness are analysed and temperature was found to be the fundamental driver. This study extends “the time–temperature stability field of Hein et al. (1978)” for the onset of opal-CT precipitation more generally. Active silica diagenesis persists over a long period of time (>35 m.y.) at low temperatures (55 °C) regardless of burial depth. Geothermal gradients and sediment accumulation rates are the principal controls of the rate of silica diagenesis transformation. Sites 794 and 795 in the Japan Sea, as typical cases of deep-sea boreholes capturing active transitions, were selected for further understanding the temperature–time control on silica diagenesis. The reconstructed thermal evolution of sediments from these representative sites demonstrates that the rapid increase in temperatures from the Late Miocene onwards occurred in response to high accumulation rates in the opal-A interval. The higher burial temperatures achieved for the opal-A sediment under higher sedimentation rates and a steeper thermal gradient led to an earlier transition at Site 795 (~5 m.y.) as compared to Site 794 (~8 m.y.). Kinetic-based models are formulated to illustrate the time–temperature dependence of biogenic silica diagenesis. One of the main findings based on this model is that although opal-CT precipitation reaches its peak levels across the transition zone at 8 and 5 m.y. after initial deposition of opal-A at Sites 794 and 795, respectively, the transformation is not completed until 14 m.y. at Site 794 and ca. 9.5 m.y. at Site 795 after initial sedimentation. Given the temperature and elapsed time, the model allows the transformation state of opal-containing sediments to be successfully predicted at any depth in these two sites

Arrested versus active silica diagenesis reaction boundaries—A review of seismic diagnostic criteria

This paper evaluates previously proposed diagnostic criteria that can be used to determine whether or not there is active migration of the opal-A to opal-CT transition zone (TZA/CT). The criteria are based on the interpretation of 2D and 3D seismic surveys and are therefore geometrical. They involve an assessment of the relationship of the TZA/CT&nbsp;with polygonal fault systems, differential compaction structures and tectonic folds. The most robust evidence for an inactive &lsquo;reaction front&rsquo; between opal-A and opal-CT bearing sediments is the discordance of the TZA/CT&nbsp;relative to present-day isotherms. Any of these may be persuasive as diagnostic criteria for the upward arrest of the diagenetic transformation at a regional scale, but actual truncation of the TZA/CT&nbsp;at the modern seabed is definitive for arrested diagenesis. This study argues that diagenetic assessment based solely on a single criterion independently is not reliable as an indicator for the current state of a silica transition. As a conclusion, the analysed seismic/structural criteria should be synthesised to provide a more credible interpretation for silica diagenesis. The use of modern 2D and 3D seismic data for the reconstruction of the diagenetic history of opaline silica bearing sediments offers a new approach to the study of silica diagenesis at a regional scale.</p

[en] BUSINESS ROLE IN WATERSHEDS: MOTIVATIONS, BENEFITS AND LIMITATIONS

Gene silencing, via RNA interference (RNAi) technologies using small interfering RNA (siRNA), has been developed as an important tool for target identification and validation in drug discovery and has huge therapeutic potential. However, effective delivery into cells presents a major challenge to the use of siRNA. pH responsive cell-penetrating peptides have attracted considerable attention in recent years as delivery vectors due to their ability to transport their cargos across the biological membrane and/or to promote endosomal escape and prevent lyososomal degradation. To evaluate the in vitro transfection efficiency of the pH responsive peptide-based siRNA delivery system, the western blotting technique is commonly employed. This method offers a simple, efficient and economical way to study the gene silencing effect of the siRNA by analysing the protein of interest in a sample with minimum equipment requirement. This chapter provides a description of siRNA delivery and analysis by western blotting protocols for qualitatively and quantitatively assessing gene silencing efficiency and selectivity