Translocation of structured polynucleotides through nanopores

We investigate theoretically the translocation of structured RNA/DNA molecules through narrow pores which allow single but not double strands to pass. The unzipping of basepaired regions within the molecules presents significant kinetic barriers for the translocation process. We show that this circumstance may be exploited to determine the full basepairing pattern of polynucleotides, including RNA pseudoknots. The crucial requirement is that the translocation dynamics (i.e., the length of the translocated molecular segment) needs to be recorded as a function of time with a spatial resolution of a few nucleotides. This could be achieved, for instance, by applying a mechanical driving force for translocation and recording force-extension curves (FEC's) with a device such as an atomic force microscope or optical tweezers. Our analysis suggests that with this added spatial resolution, nanopores could be transformed into a powerful experimental tool to study the folding of nucleic acids.Comment: 9 pages, 5 figure

Forests and water: a state-of-the-art review for Colorado

Includes bibliographical references (pages 65-75).Forests occupy 22.6 million acres in Colorado, or 32 percent of the land area, and nearly three-quarters of the forest lands in Colorado are in public ownership. About 55 percent of the forested area is considered suitable for forest harvest. National forests comprise nearly half of the forested area and approximately 60 percent of the area is considered suitable for forest harvest. There are no significant, privately-owned, industrial forest lands in Colorado. Historic photographs, forest stand records, and other data indicate that forest density in Colorado is generally greater than in the mid to late 1800s. This increase in forest density, attributed to suppression of forest fires, reduced grazing, and lower rates of forest harvest for timber, fuel, and other products, are generally believed to have decreased annual water yields. Annual water yields from the 1.34 million acres of national forest lands in the North Platte River basin are estimated to have decreased by approximately 8 to 14 percent or 135,000 to 185,000 acre-feet per year, depending on the assumed stand history for the spruce-fir forests. Hydrologic models indicate that average annual water yields could be increased in the North Platte River basin by about 55,000 acre-feet per year if all 502,000 acres designated as suitable for timber harvest were regularly harvested on a sustained yield basis. Similar data are not available for other river basins in Colorado, although the overall trends are probably similar. This research looked at how reducing forest canopy affects the rate of spring snowmelt and water yield, how it affects evapotranspiration, what happens when the forest regrows, whether reducing forest density affects water yields if annual precipitation is a factor, the effects on water quality, and the necessity for water storage facilities to store the increased runoff. The report does not attempt to address the myriad of other issues that must be considered when evaluating various management alternatives for forested lands. Some of these issues include the numerous laws and regulations that affect land management, economic considerations, the downstream uses of water and water storage capacities, and the effects of forest management on recreation, local communities, aesthetics, and other plant and animal species.Sponsored by: Colorado River Water Conservation District, Colorado Water Resources Research Institute, Denver Water, Northern Colorado Water Conservancy District and financed in part by the U.S. Department of the Interior, Geological Survey, through the Colorado Water Resources Research Institute and Grant no. 01HQGR0077

Bending and looping of long DNA by Polycomb repressive complex 2 revealed by AFM imaging in liquid

Polycomb repressive complex 2 (PRC2) is a histone methyltransferase that methylates histone H3 at Lysine 27. PRC2 is critical for epigenetic gene silencing, cellular differentiation and the formation of facultative heterochromatin. It can also promote or inhibit oncogenesis. Despite this importance, the molecular mechanisms by which PRC2 compacts chromatin are relatively understudied. Here, we visualized the binding of PRC2 to naked DNA in liquid at the single-molecule level using atomic force microscopy. Analysis of the resulting images showed PRC2, consisting of five subunits (EZH2, EED, SUZ12, AEBP2&nbsp;and RBBP4), bound to a 2.5-kb DNA with an apparent dissociation constant (⁠KappD⁠) of 150 &plusmn; 12 nM. PRC2 did not show sequence-specific binding to a region of high GC content (76%) derived from a CpG island embedded in such a long DNA substrate. At higher concentrations, PRC2 compacted DNA by forming DNA loops typically anchored by two or more PRC2 molecules. Additionally, PRC2 binding led to a 3-fold increase in the local bending of DNA&rsquo;s helical backbone without evidence of DNA wrapping around the protein. We suggest that the bending and looping of DNA by PRC2, independent of PRC2&rsquo;s methylation activity, may contribute to heterochromatin formation and therefore epigenetic gene silencing.</p

Tsunami-Related Data: A Review of Available Repositories Used in Scientific Literature

Various organizations and institutions store large volumes of tsunami-related data, whose availability and quality should benefit society, as it improves decision making before the tsunami occurrence, during the tsunami impact, and when coping with the aftermath. However, the existing digital ecosystem surrounding tsunami research prevents us from extracting the maximum benefit from our research investments. The main objective of this study is to explore the field of data repositories providing secondary data associated with tsunami research and analyze the current situation. We analyze the mutual interconnections of references in scientific studies published in the Web of Science database, governmental bodies, commercial organizations, and research agencies. A set of criteria was used to evaluate content and searchability. We identified 60 data repositories with records used in tsunami research. The heterogeneity of data formats, deactivated or nonfunctional web pages, the generality of data repositories, or poor dataset arrangement represent the most significant weak points. We outline the potential contribution of ontology engineering as an example of computer science methods that enable improvements in tsunami-related data management

Pharmacologic IRE1/XBP1s Activation Confers Targeted ER Proteostasis Reprogramming

Activation of the IRE1/XBP1s signaling arm of the unfolded protein response (UPR) is a promising strategy to correct defects in endoplasmic reticulum (ER) proteostasis implicated in diverse diseases. However, no pharmacologic activators of this pathway identified to date are suitable for ER proteostasis remodeling through selective activation of IRE1/XBP1s signaling. Here, we use high-throughput screening to identify non-toxic compounds that induce ER proteostasis remodeling through IRE1/XBP1s activation. We employ transcriptional profiling to stringently confirm that our prioritized compounds selectively activate IRE1/XBP1s signaling without activating other cellular stress-responsive signaling pathways. Furthermore, we demonstrate that our compounds improve ER proteostasis of destabilized variants of amyloid precursor protein (APP) through an IRE1-dependent mechanism and reduce APP-associated mitochondrial toxicity in cellular models. These results establish highly selective IRE1/XBP1s activating compounds that can be widely employed to define the functional importance of IRE1/XBP1s activity for ER proteostasis regulation in the context of health and disease. [Figure not available: see fulltext.]

DHODH modulates transcriptional elongation in the neural crest and melanoma

Melanoma is a tumour of transformed melanocytes, which are originally derived from the embryonic neural crest. It is unknown to what extent the programs that regulate neural crest development interact with mutations in the BRAF oncogene, which is the most commonly mutated gene in human melanoma1. We have used zebrafish embryos to identify the initiating transcriptional events that occur on activation of human BRAF(V600E) (which encodes an amino acid substitution mutant of BRAF) in the neural crest lineage. Zebrafish embryos that are transgenic for mitfa:BRAF(V600E) and lack p53 (also known as tp53) have a gene signature that is enriched for markers of multipotent neural crest cells, and neural crest progenitors from these embryos fail to terminally differentiate. To determine whether these early transcriptional events are important for melanoma pathogenesis, we performed a chemical genetic screen to identify small-molecule suppressors of the neural crest lineage, which were then tested for their effects on melanoma. One class of compound, inhibitors of dihydroorotate dehydrogenase (DHODH), for example leflunomide, led to an almost complete abrogation of neural crest development in zebrafish and to a reduction in the self-renewal of mammalian neural crest stem cells. Leflunomide exerts these effects by inhibiting the transcriptional elongation of genes that are required for neural crest development and melanoma growth. When used alone or in combination with a specific inhibitor of the BRAF(V600E) oncogene, DHODH inhibition led to a marked decrease in melanoma growth both in vitro and in mouse xenograft studies. Taken together, these studies highlight developmental pathways in neural crest cells that have a direct bearing on melanoma formation

The origin of large molecules in primordial autocatalytic reaction networks

Large molecules such as proteins and nucleic acids are crucial for life, yet their primordial origin remains a major puzzle. The production of large molecules, as we know it today, requires good catalysts, and the only good catalysts we know that can accomplish this task consist of large molecules. Thus the origin of large molecules is a chicken and egg problem in chemistry. Here we present a mechanism, based on autocatalytic sets (ACSs), that is a possible solution to this problem. We discuss a mathematical model describing the population dynamics of molecules in a stylized but prebiotically plausible chemistry. Large molecules can be produced in this chemistry by the coalescing of smaller ones, with the smallest molecules, the `food set', being buffered. Some of the reactions can be catalyzed by molecules within the chemistry with varying catalytic strengths. Normally the concentrations of large molecules in such a scenario are very small, diminishing exponentially with their size. ACSs, if present in the catalytic network, can focus the resources of the system into a sparse set of molecules. ACSs can produce a bistability in the population dynamics and, in particular, steady states wherein the ACS molecules dominate the population. However to reach these steady states from initial conditions that contain only the food set typically requires very large catalytic strengths, growing exponentially with the size of the catalyst molecule. We present a solution to this problem by studying `nested ACSs', a structure in which a small ACS is connected to a larger one and reinforces it. We show that when the network contains a cascade of nested ACSs with the catalytic strengths of molecules increasing gradually with their size (e.g., as a power law), a sparse subset of molecules including some very large molecules can come to dominate the system.Comment: 49 pages, 17 figures including supporting informatio

Experience and Challenges from Clinical Trials with Malaria Vaccines in Africa.

Malaria vaccines are considered amongst the most important modalities for potential elimination of malaria disease and transmission. Research and development in this field has been an area of intense effort by many groups over the last few decades. Despite this, there is currently no licensed malaria vaccine. Researchers, clinical trialists and vaccine developers have been working on many approached to make malaria vaccine available.African research institutions have developed and demonstrated a great capacity to undertake clinical trials in accordance to the International Conference on Harmonization-Good Clinical Practice (ICH-GCP) standards in the last decade; particularly in the field of malaria vaccines and anti-malarial drugs. This capacity is a result of networking among African scientists in collaboration with other partners; this has traversed both clinical trials and malaria control programmes as part of the Global Malaria Action Plan (GMAP). GMAP outlined and support global strategies toward the elimination and eradication of malaria in many areas, translating in reduction in public health burden, especially for African children. In the sub-Saharan region the capacity to undertake more clinical trials remains small in comparison to the actual need.However, sustainability of the already developed capacity is essential and crucial for the evaluation of different interventions and diagnostic tools/strategies for other diseases like TB, HIV, neglected tropical diseases and non-communicable diseases. There is urgent need for innovative mechanisms for the sustainability and expansion of the capacity in clinical trials in sub-Saharan Africa as the catalyst for health improvement and maintained