Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase.

Nucleic Acid Therapeutics (NATs), including siRNAs and AntiSense Oligonucleotides (ASOs), have great potential to drug the undruggable genome. Targeting siRNAs and ASOs to specific cell types of interest has driven dramatic improvement in efficacy and reduction in toxicity. Indeed, conjugation of tris-GalNAc to siRNAs and ASOs has shown clinical efficacy in targeting diseases driven by liver hepatocytes. However, targeting non-hepatic diseases with oligonucleotide therapeutics has remained problematic for several reasons, including targeting specific cell types and endosomal escape. Monoclonal antibody (mAb) targeting of siRNAs and ASOs has the potential to deliver these drugs to a variety of specific cell and tissue types. However, most conjugation strategies rely on random chemical conjugation through lysine or cysteine residues resulting in conjugate heterogeneity and a distribution of Drug:Antibody Ratios (DAR). To produce homogeneous DAR-2 conjugates with two siRNAs per mAb, we developed a novel two-step conjugation procedure involving microbial transglutaminase (MTGase) tagging of the antibody C-terminus with an azide-functionalized linker peptide that can be subsequently conjugated to dibenzylcyclooctyne (DBCO) bearing oligonucleotides through azide-alkyne cycloaddition. Antibody-siRNA (and ASO) conjugates (ARCs) produced using this strategy are soluble, chemically defined targeted oligonucleotide therapeutics that have the potential to greatly increase the number of targetable cell types

Effect of mattress deflection on CPR quality assessment for older children and adolescents

Appropriate chest compression (CC) depth is associated with improved CPR outcome. CCs provided in hospital are often conducted on a compliant mattress. The objective was to quantify the effect of mattress compression on the assessment of CPR quality in children. Methods: A force and deflection sensor (FDS) was used during CPR in the Pediatric Intensive Care Unit and Emergency Department of a children's hospital. The sensor was interposed between the chest of the patient and hands of the rescuer and measured CC depth. Following CPR event, each event was reconstructed with a manikin and an identical mattress/backboard/patient configuration. CCs were performed using FDS on the sternum and a reference accelerometer attached to the spine of the manikin, providing a means to Calculate the mattress deflection. Results: Twelve CPR events with 14,487 CC (11 patients, median age 14.9 years) were recorded and reconstructed: 9 on ICU beds (9296 CC), 3 on stretchers (5191 CC). Measured mean CC depth during CPR was 47 +/- 8 mm on ICU beds, and 45 +/- 7 mm on stretcher beds with overestimation of 13 +/- 4 mm and 4 +/- 1 mm, respectively, due to mattress compression. After adjusting for this, the proportion of CC that met the CPR guidelines decreased from 88.4 to 31.8% on ICU beds (p < 0.001), and 86.3 to 64.7% on stretcher (p < 0.001 The proportion of appropriate depth CC was significantly smaller on ICU beds (p < 0.001). Conclusion: CC conducted on a non-rigid surface may not be deep enough. FDS may overestimate CC depth by 28% on ICU beds, and 10% on stretcher beds

Urban impacts on a prairie groundwater system : estimation of anthropogenic contributions of water and potential effects on water table development

xiii, 190 leaves : ill., maps ; 28 cm.In subhumid to arid climates throughout the world, recharge to groundwater in urban areas is often found to be higher than pre-urbanization rates, despite an increased percentage of impermeable surfaces. Groundwater recharge in the city of Lethbridge is substantially higher than recharge rates prior to urbanization, resulting in the formation of perched water table conditions. High perched water table conditions, typically at depths between one and 2.5 metres, have created problems for the City and University of Lethbridge, including the increased occurrence of slope failures along nearby coulees. This study estimates of the volume of excess water available for groundwater recharge through the practices of urban turfgrass irrigation, and water storage. Between May and September, 1990-1996 irrigation was applied far above evapotranspiration demands, resulting in large volumes of water available for groundwater recharge in the Varsity Village subdivision of the City of Lethbridge. The relationship between the amount of water applied and the development of perched water table systems was strong enough that equations between inputs and water table depth could be derived, and used to predict water table elevation

Analysis of Soil Freeze/Thaw Signatures During Slapex F/T Campaign

Permanently frozen and seasonally frozen soils occur over a large portion of the Earth's land surface. Changes in the freeze/thaw state of the land surface reflects major changes in thermal and hydraulic properties as well as acting as a "switch" for many ecological processes. In short, soil freeze/thaw state is a fundamental land surface variable in the water and energy cycles, and it connects to the carbon cycle. Surface freeze/thaw state is observable by passive and active microwave sensors. For example, NASA's Soil Moisture Active Passive (SMAP) mission includes a freeze/thaw data product. Such satellite sensing offers routine all-season and all-weather global observations of soil freeze/thaw state with the application of suitable algorithms. We describe early finding from the SLAPex Freeze/Thaw campaign, believed to be the first airborne campaign of its type, focusing on soil freeze/thaw

Canadian Experiment for Soil Moisture in 2010 (CanEX-SM10): Overview and Preliminary Results

The Canadian Experiment for Soil Moisture in 2010 (CanEx-SM10) was carried out in Saskatchewan, Canada from 31 May to 16 June, 2010. Its main objective was to contribute to Soil Moisture and Ocean salinity (SMOS) mission validation and the pre-launch assessment of Soil Moisture and Active and Passive (SMAP) mission. During CanEx-SM10, SMOS data as well as other passive and active microwave measurements were collected by both airborne and satellite platforms. Ground-based measurements of soil (moisture, temperature, roughness, bulk density) and vegetation characteristics (Leaf Area Index, biomass, vegetation height) were conducted close in time to the airborne and satellite acquisitions. Besides, two ground-based in situ networks provided continuous measurements of meteorological conditions and soil moisture and soil temperature profiles. Two sites, each covering 33 km x 71 km (about two SMOS pixels) were selected in agricultural and boreal forested areas in order to provide contrasting soil and vegetation conditions. This paper describes the measurement strategy, provides an overview of the data sets and presents preliminary results. Over the agricultural area, the airborne L-band brightness temperatures matched up well with the SMOS data. The Radio frequency interference (RFI) observed in both SMOS and the airborne L-band radiometer data exhibited spatial and temporal variability and polarization dependency. The temporal evolution of SMOS soil moisture product matched that observed with the ground data, but the absolute soil moisture estimates did not meet the accuracy requirements (0.04 m3/m3) of the SMOS mission. AMSR-E soil moisture estimates are more closely correlated with measured soil moisture

Ensemble Identification of Spectral Bands Related to Soil Organic Carbon Levels over an Agricultural Field in Southern Ontario, Canada

The recent use of hyperspectral remote sensing imagery has introduced new opportunities for soil organic carbon (SOC) assessment and monitoring. These data enable monitoring of a wide variety of soil properties but pose important methodological challenges. Highly correlated hyperspectral spectral bands can affect the prediction and accuracy as well as the interpretability of the retrieval model. Therefore, the spectral dimension needs to be reduced through a selection of specific spectral bands or regions that are most helpful to describing SOC. This study evaluates the efficiency of visible near-infrared (VNIR) and shortwave near-infrared (SWIR) hyperspectral data to identify the most informative hyperspectral bands responding to SOC content in agricultural soils. Soil samples (111) were collected over an agricultural field in southern Ontario, Canada and analyzed against two hyperspectral datasets: An airborne Nano-Hyperspec imaging sensor with 270 bands (400–1000 nm) and a laboratory hyperspectral dataset (ASD FieldSpec 3) along the 1000–2500 nm range (NIR-SWIR). In parallel, a multimethod modeling approach consisting of random forest, support vector machine, and partial least squares regression models was used to conduct band selections and to assess the validity of the selected bands. The multimethod model resulted in a selection of optimal band or regions over the VNIR and SWIR sensitive to SOC and potentially for mapping. The bands that achieved the highest respective importance values were 711–715, 727, 986–998, and 433–435 nm regions (VNIR); and 2365–2373, 2481–2500, and 2198–2206 nm (NIR-SWIR). Some of these bands are in agreement with the absorption features of SOC reported in the literature, whereas others have not been reported before. Ultimately, the selection of optimal band and regions is of importance for quantification of agricultural SOC and would provide a new framework for creating optimized SOC-specific sensors

A Combined Linkage and Exome Sequencing Analysis for Electrocardiogram Parameters in the Erasmus Rucphen Family Study

Electrocardiogram (ECG) measurements play a key role in the diagnosis and prediction of cardiac arrhythmias and sudden cardiac death. ECG parameters, such as the PR, QRS, and QT intervals, are known to be heritable and genome-wide association studies of these phenotypes have been successful in identifying common variants; however, a large proportion of the genetic variability of these traits remains to be elucidated. The aim of this study was to discover loci potentially harboring rare variants utilizing variance component linkage analysis in 1547 individuals from a large family-based study, the Erasmus Rucphen Family Study (ERF). Linked regions were further explored using exome sequencing. Five suggestive linkage peaks were identified: two for QT interval (1q24, LOD = 2.63; 2q34, LOD = 2.05), one for QRS interval (1p35, LOD = 2.52) and two for PR interval (9p22, LOD = 2.20; 14q11, LOD = 2.29). Fine-mapping using exome sequence data identified a C > G missense variant (c.713C > G, p.Ser238Cys) in the FCRL2 gene associated with QT (rs74608430; P = 2.8 x 10(-4), minor allele frequency = 0.019). Heritability analysis demonstrated that the SNP explained 2.42% of the trait's genetic variability in ERF (P = 0.02). Pathway analysis suggested that the gene is involved in cytosolic Ca2+ levels (P = 3.3 x 10(-3)) and AMPK stimulated fatty acid oxidation in muscle (P = 4.1 x 10(-3)). Look-ups in bioinformatics resources showed that expression of FCRL2 is associated with ARHGAP24 and SETBP1 expression. This finding was not replicated in the Rotterdam study. Combining the bioinformatics information with the association and linkage analyses, FCRL2 emerges as a strong candidate gene for QT interval

A combined linkage and exome sequencing analysis for electrocardiogram parameters in the erasmus rucphen family study

Electrocardiogram (ECG) measurements play a key role in the diagnosis and prediction of cardiac arrhythmias and sudden cardiac death. ECG parameters, such as the PR, QRS, and QT intervals, are known to be heritable and genome-wide association studies of these phenotypes have been successful in identifying common variants; however, a large proportion of the genetic variability of these traits remains to be elucidated. The aim of this study was to discover loci potentially harboring rare variants utilizing variance component linkage analysis in 1547 individuals from a large family-based study, the Erasmus Rucphen Family Study (ERF). Linked regions were further explored using exome sequencing. Five suggestive linkage peaks were identified: two for QT interval (1q24, LOD = 2.63; 2q34, LOD = 2.05), one for QRS interval (1p35, LOD = 2.52) and two for PR interval (9p22, LOD = 2.20; 14q11, LOD = 2.29). Fine-mapping using exome sequence data identified a C &gt; G missense variant (c.713C &gt; G, p.Ser238Cys) in the FCRL2 gene associated with QT (rs74608430; P = 2.8 × 10-4, minor allele frequency = 0.019). Heritability analysis demonstrated that the SNP explained 2.42% of the trait's genetic variability in ERF (P = 0.02). Pathway analysis suggested that the gene is involved in cytosolic Ca2+ levels (P = 3.3 × 10-3) and AMPK stimulated fatty acid oxidation in muscle (P = 4.1 × 10-3). Look-ups in bioinformatics resources showed that expression of FCRL2 is associated with ARHGAP24 and SETBP1 expression. This finding was not replicated in the Rotterdam study. Combining the bioinformatics information with the association and linkage analyses, FCRL2 emerges as a strong candidate gene for QT interval. © 2016 Silva, Zorkoltseva, Amin, Demirkan, van Leeuwen, Kors, van den Berg, Stricker, Uitterlinden, Kirichenko, Witteman, Willemsen, Oostra, Axenovich, van Duijn and Isaacs

Using [Ne V]/[Ne III] to Understand the Nature of Extreme-Ionization Galaxies

Spectroscopic studies of extreme-ionization galaxies (EIGs) are critical to our understanding of exotic systems throughout cosmic time. These EIGs exhibit spectral features requiring >54.42 eV photons: the energy needed to fully ionize helium into He2+ and emit He II recombination lines. They are likely key contributors to reionization, and they can also probe exotic stellar populations or accretion onto massive black holes. To facilitate the use of EIGs as probes of high ionization, we focus on ratios constructed from strong rest-frame UV/optical emission lines, specifically [O III] 5008, H-beta, [Ne III] 3870, [O II] 3727,3729, and [Ne V] 3427. These lines probe the relative intensity at energies of 35.12, 13.62, 40.96, 13.62 eV, and 97.12, respectively, covering a wider range of ionization than traced by other common rest-frame UV/optical techniques. We use ratios of these lines ([Ne V]/[Ne III] = Ne53 and [Ne III]/[O II]), which are closely separated in wavelength, and mitigates effects of dust attenuation and uncertainties in flux calibration. We make predictions from photoionization models constructed from Cloudy that use a broad range of stellar populations and black hole accretion models to explore the sensitivity of these line ratios to changes in the ionizing spectrum. We compare our models to observations from the Hubble Space Telescope and James Webb Space Telescope of galaxies with strong high-ionization emission lines at z ~ 0, z ~ 2, and z ~ 7. We show that the Ne53 ratio can separate galaxies with ionization from 'normal' stellar populations from those with AGN and even 'exotic' Population III models. We introduce new selection methods to identify galaxies with photoionization driven by Population III stars or intermediate-mass black hole accretion disks that could be identified in upcoming high-redshift spectroscopic surveys.Comment: 16 pages, 5 figures, 1 table. Accepted in Ap

Structural mechanism underpinning cross-reactivity of a CD8(+) T-cell clone that recognizes a peptide derived from human telomerase reverse transcriptase

T-cell cross-reactivity is essential for effective immune surveillance but has also been implicated as a pathway to autoimmunity. Previous studies have demonstrated that T-cell receptors (TCRs) that focus on a minimal motif within the peptide are able to facilitate a high level of T-cell cross-reactivity. However, the structural database shows that most TCRs exhibit less focused antigen binding involving contact with more peptide residues. To further explore the structural features that allow the clonally expressed TCR to functionally engage with multiple peptide-major histocompatibility complexes (pMHCs), we examined the ILA1 CD8(+) T-cell clone that responds to a peptide sequence derived from human telomerase reverse transcriptase. The ILA1 TCR contacted its pMHC with a broad peptide binding footprint encompassing spatially distant peptide residues. Despite the lack of focused TCR-peptide binding, the ILA1 T-cell clone was still cross-reactive. Overall, the TCR-peptide contacts apparent in the structure correlated well with the level of degeneracy at different peptide positions. Thus, the ILA1 TCR was less tolerant of changes at peptide residues that were at, or adjacent to, key contact sites. This study provides new insights into the molecular mechanisms that control T-cell cross-reactivity with important implications for pathogen surveillance, autoimmunity, and transplant rejection