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1.
Lab Chip ; 23(2): 285-294, 2023 01 17.
Article in English | MEDLINE | ID: mdl-36524732

ABSTRACT

Ribonucleoproteins (RNPs), particularly microRNA-induced silencing complex (miRISC), have been associated with cancer-related gene regulation. Specific RNA-protein associations in miRISC complexes or those found in let-7 lin28A complexes can downregulate tumor-suppressing genes and can be directly linked to cancer. The high protein-RNA electrostatic binding affinity is a particular challenge for the quantification of the associated microRNAs (miRNAs). We report here the first microfluidic point-of-care assay that allows direct quantification of RNP-associated RNAs, which has the potential to greatly advance RNP profiling for liquid biopsy. Key to the technology is an integrated cation-anion exchange membrane (CEM/AEM) platform for rapid and irreversible dissociation (k = 0.0025 s-1) of the RNP (Cas9-miR-21) complex and quantification of its associated miR-21 in 40 minutes. The CEM-induced depletion front is used to concentrate the RNP at the depletion front such that the high electric field (>100 V cm-1) within the concentration boundary layer induces irreversible dissociation of the low KD (∼0.5 nM) complex, with ∼100% dissociation even though the association rate (kon = 6.1 s-1) is 1000 times higher. The high field also electrophoretically drives the dissociated RNA out of the concentrated zone without reassociation. A detection limit of 1.1 nM is achieved for Cy3 labelled miR-21.


Subject(s)
MicroRNAs , Microfluidics , Neoplasms , Humans , Gene Expression Regulation , Microfluidics/instrumentation , MicroRNAs/chemistry , Ribonucleoproteins/chemistry
2.
J Anim Ecol ; 88(12): 1873-1887, 2019 12.
Article in English | MEDLINE | ID: mdl-31330569

ABSTRACT

Urban areas affect terrestrial ecological processes and local weather, but we know little about their effect on aerial ecological processes. Here, we identify urban from non-urban areas based on the intensity of artificial light at night (ALAN) in the landscape, and, along with weather covariates, evaluate the effect of urbanization on flight altitudes of nocturnally migrating birds. Birds are attracted to ALAN; hence, we predicted that altitudes would be lower over urban than over non-urban areas. However, other factors associated with urbanization may also affect flight altitudes. For example, surface temperature and terrain roughness are higher in urban areas, increasing air turbulence and height of the boundary layer, and affecting local winds. We used data from nine weather surveillance radars in the eastern United States to estimate altitudes at five quantiles of the vertical distribution of birds migrating at night over urban and non-urban areas during five consecutive spring and autumn migration seasons. We fit Generalized Linear Mixed Models by season for each of the five quantiles of bird flight altitude and their differences between urban and non-urban areas. After controlling for other environmental variables and contrary to our prediction, we found that birds generally fly higher over urban areas compared to rural areas in spring, and marginally higher at the mid-layers of the vertical distribution in autumn. We also identified a small interaction effect between urbanization and crosswind speed, and between urbanization and surface air temperature, on flight altitudes. We also found that the difference in flight altitudes of nocturnally migrating birds between urban and non-urban areas varied among radars and seasons, but was consistently higher over urban areas throughout the years sampled. Our results suggest that the effects of urbanization on wildlife extend into the aerosphere and are complex, stressing the need of understanding the influence of anthropogenic factors on airspace habitat.


Subject(s)
Altitude , Animal Migration , Animals , Birds , Flight, Animal , Weather , Wind
3.
Conserv Biol ; 26(1): 68-77, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22010858

ABSTRACT

Predictive models of species distributions are typically developed with data collected along roads. Roadside sampling may provide a biased (nonrandom) sample; however, it is currently unknown whether roadside sampling limits the accuracy of predictions generated by species distribution models. We tested whether roadside sampling affects the accuracy of predictions generated by species distribution models by using a prospective sampling strategy designed specifically to address this issue. We built models from roadside data and validated model predictions at paired locations on unpaved roads and 200 m away from roads (off road), spatially and temporally independent from the data used for model building. We predicted species distributions of 15 bird species on the basis of point-count data from a landbird monitoring program in Montana and Idaho (U.S.A.). We used hierarchical occupancy models to account for imperfect detection. We expected predictions of species distributions derived from roadside-sampling data would be less accurate when validated with data from off-road sampling than when it was validated with data from roadside sampling and that model accuracy would be differentially affected by whether species were generalists, associated with edges, or associated with interior forest. Model performance measures (kappa, area under the curve of a receiver operating characteristic plot, and true skill statistic) did not differ between model predictions of roadside and off-road distributions of species. Furthermore, performance measures did not differ among edge, generalist, and interior species, despite a difference in vegetation structure along roadsides and off road and that 2 of the 15 species were more likely to occur along roadsides. If the range of environmental gradients is surveyed in roadside-sampling efforts, our results suggest that surveys along unpaved roads can be a valuable, unbiased source of information for species distribution models.


Subject(s)
Birds , Conservation of Natural Resources/methods , Animals , Geographic Information Systems , Idaho , Models, Statistical , Montana , Population Density
4.
Blood ; 102(13): 4393-8, 2003 Dec 15.
Article in English | MEDLINE | ID: mdl-12933577

ABSTRACT

Intravenous administration of recombinant human factor IX (rhFIX) acutely corrects the coagulopathy in hemophilia B dogs. To date, 20 of 20 dogs developed inhibitory antibodies to the xenoprotein, making it impossible to determine if new human FIX products, formulations, or methods of chronic administration can reduce bleeding frequency. Our goal was to determine whether hemophilia B dogs rendered tolerant to rhFIX would have reduced bleeding episodes while on sustained prophylactic rhFIX administered subcutaneously. Reproducible methods were developed for inducing tolerance to rhFIX in this strain of hemophilia B dogs, resulting in a significant reduction in the development of inhibitors relative to historical controls (5 of 12 versus 20 or 20, P <.001). The 7 of 12 tolerized hemophilia B dogs exhibited shortened whole blood clotting times (WBCTs), sustained detectable FIX antigen, undetectable Bethesda inhibitors, transient or no detectable antihuman FIX antibody titers by enzyme-linked immunosorbent assay (ELISA), and normal clearance of infused rhFIX. Tolerized hemophilia B dogs had 69% reduction in bleeding frequency in year 1 compared with nontolerized hemophilia B dogs (P =.0007). If proven safe in human clinical trials, subcutaneous rhFIX may provide an alternate approach to prophylactic therapy in selected patients with hemophilia B.


Subject(s)
Factor IX/therapeutic use , Hemophilia B/drug therapy , Hemorrhage/prevention & control , Animals , Animals, Inbred Strains , Animals, Newborn , Antibodies, Heterophile/biosynthesis , Antibodies, Heterophile/immunology , Disease Models, Animal , Dogs , Factor IX/administration & dosage , Factor IX/immunology , Hemophilia B/complications , Hemorrhage/etiology , Humans , Immune Tolerance , Injections, Subcutaneous , Male , Pilot Projects , Recombinant Proteins/administration & dosage , Recombinant Proteins/immunology , Recombinant Proteins/therapeutic use , Reproducibility of Results
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