Small sized EGFR1 and HER2 specific bifunctional antibody for targeted cancer therapy

licenses/by-nc-nd/3.0/). Reproduction is permitted for personal, noncommercial use, provided that the article is in whole, unmodified, and properly cited. Received: 2014.07.10; Accepted: 2015.01.05; Published: 2015.01.21 Targeting tumors using miniature antibodies is a novel and attractive therapeutic approach, as these biomolecules exhibit low immunogenicity, rapid clearance, and high targeting specificity. However, most of the small-sized antibodies in existence do not exhibit marked anti-tumor ef-fects, which limit their use in targeted cancer immunotherapy. To overcome this difficulty in targeting multiple biomarkers by combination therapies, we designed a new bifunctional antibody, named MaAbNA (multivalent antibody comprised of nanobody and affibody moieties), capable of targeting EGFR1 and HER2, which are widely overexpressed in a variety of tumor types. The small-sized (29 kDa) MaAbNA, which was expressed in E.coli, consists of one anti-EGFR1 nano-body and two anti-HER2 affibodies, and possesses high affinity (KD) for EGFR1 (~4.1 nM) and HER2 (~4.7 nM). In order to enhance its anti-tumor activity, MaAbNA was conjugated with adriamycin (ADM) using a PEG2000 linker, forming a new complex anticancer drug, MaAbNA-PEG2000-ADM. MaAbNA exhibited high inhibitory effects on tumor cell

Fine mapping of a QTL for ear size on porcine chromosome 5 and identification of high mobility group AT-hook 2 (HMGA2) as a positional candidate gene

<p>Abstract</p> <p>Background</p> <p>Ear size and shape are distinct conformation characteristics of pig breeds. Previously, we identified a significant quantitative trait locus (QTL) influencing ear surface on pig chromosome 5 in a White Duroc × Erhualian F₂resource population. This QTL explained more than 17% of the phenotypic variance.</p> <p>Methods</p> <p>Four new markers on pig chromosome 5 were genotyped across this F₂population. RT-PCR was performed to obtain expression profiles of different candidate genes in ear tissue. Standard association test, marker-assisted association test and F-drop test were applied to determine the effects of single nucleotide polymorphisms (SNP) on ear size. Three synthetic commercial lines were also used for the association test.</p> <p>Results</p> <p>We refined the QTL to an 8.7-cM interval and identified three positional candidate genes i.e. <it>HMGA2</it>, <it>SOX5 </it>and <it>PTHLH </it>that are expressed in ear tissue. Seven SNP within these three candidate genes were selected and genotyped in the F₂population. Of the seven SNP, <it>HMGA2 </it>SNP (JF748727: g.2836 A > G) showed the strongest association with ear size in the standard association test and marker-assisted association test. With the F-drop test, F value decreased by more than 97% only when the genotypes of <it>HMGA2 </it>g.2836 A > G were included as a fixed effect. Furthermore, the significant association between g.2836 A > G and ear size was also demonstrated in the synthetic commercial Sutai pig line. The haplotype-based association test showed that the phenotypic variance explained by <it>HMGA2 </it>was similar to that explained by the QTL and at a much higher level than by <it>SOX5</it>. More interestingly, <it>HMGA2 </it>is also located within the dog orthologous chromosome region, which has been shown to be associated with ear type and size.</p> <p>Conclusions</p> <p><it>HMGA2 </it>was the closest gene with a potential functional effect to the QTL or marker for ear size on chromosome 5. This study will contribute to identify the causative gene and mutation underlying this QTL.</p

Inhibition of HIF-1α Reduced Blood Brain Barrier Damage by Regulating MMP-2 and VEGF During Acute Cerebral Ischemia

Increase of blood brain barrier (BBB) permeability after acute ischemia stroke is a predictor to intracerebral hemorrhage transformation (HT) for tissue plasminogen activator (tPA) thrombolysis and post-endovascular treatment. Previous studies showed that 2-h ischemia induced damage of BBB integrity and matrix metalloproteinase-2 (MMP-2) made major contribution to this disruption. A recent study showed that blocking β2-adrenergic receptor (β2-AR) alleviated ischemia-induced BBB injury by reducing hypoxia-inducible factor-1 alpha (HIF-1α) level. In this study, we sought to investigate the interaction of HIF-1α with MMP-2 and vascular endothelial growth factor (VEGF) in BBB injury after acute ischemia stroke. Rat suture middle cerebral artery occlusion (MCAO) model was used to mimic ischemia condition. Our results showed that ischemia produced BBB damage and MMP-2/9 upregulation was colocalized with Rhodamine-dextran leakage. Pretreatment with YC-1, a HIF-1α inhibitor, alleviated 2-h ischemia-induced BBB injury significantly accompanied by decrease of MMP-2 upregulation. In addition, YC-1 also prevented VEGF-induced BBB damage. Of note, VEGF was shown to be colocalized with neurons but not astrocytes. Taken together, BBB damage was reduced by inhibition of interaction of HIF-1α with MMP-2 and VEGF during acute cerebral ischemia. These findings provide mechanisms underlying BBB damage after acute ischemia stroke and may help reduce thrombolysis- and post-endovascular treatment-related cerebral hemorrhage

Potential Variation of Evapotranspiration Induced by Typical Vegetation Changes in Northwest China

Evapotranspiration (ET), as a key eco-hydrological parameter, plays an important role in understanding sustainable ecosystem development. Each plant category has a unique functional trait on transpiration and photosynthesis, with ET implying that water cycle and energy transformation is linked with vegetation type. Changes in surface vegetation directly alter biophysical land surface properties, hence affecting energy and ET transfer. With the rapid increase in land surface changes, there is a need to further understand and quantify the effects of vegetation change on ET, especially over the vulnerable water-cycle region in the arid and semi-arid regions of Northwest China. We adopted the GlobalLand30 land cover and MOD16A2 in 2010 and 2020 to investigate, discuss the spatio-temporal characteristics of annual and seasonal ET of cultivated land, grassland, and forests in Northwest China, and quantify the impact on vegetation changes with absolute and relative changes from different climatic subecoregions on ET. Our results show the following: (1) Forest ET was generally the highest at 688 mm, followed by cultivated land and grassland with 200&ndash;400 mm in arid climatic subecoregions. (2) Returning cultivated land to forests and cultivated land expansion potentially enhances ET by 90&ndash;110 mm/10a, with the relative rate of change increasing by 22.1% and 45.8%, respectively, away from unchanged vegetation within identical subecoregions. (3) The ET of most investigated areas gains the highest value in summer, followed by spring, autumn, and winter. This study provides reference for sustainable ecosystem development and the reasonable utilization of limited water resources in Northwest China

Angle Effect on Typical Optical Remote Sensing Indices in Vegetation Monitoring

Optical remote sensing indices play an important role in vegetation information extraction and have been widely serving ecology, agriculture and forestry, urban monitoring, and other communities. Remote sensing indices are constructed from individual bands depending on special characteristics to enhance the typical spectral features for the identification or distinction of surface land covers. With the development of quantitative remote sensing, there is a rapid increasing requirement for accurate data processing and modeling. It is well known that the geometry-induced variation observed in surface reflectance is not ignorable, but the situation of uncertainty thereby introduced into these indices still needs further detailed understanding. We adopted the ground multi-angle hyperspectrum, spectral response function (SRF) of Thematic Mapper (TM), Enhanced Thematic Mapper (ETM+), Operational Land Imager (OLI), Moderate-Resolution Imaging Spectroradiometer (MODIS), and Multi-Spectral Instrument (MSI) optical sensors and simulated their sensor-like spectral reflectance; then, we investigated the potential angle effect uncertainty on optical indices that have been frequently involved in vegetation monitoring and examined the forward/backward effect over both the ground-based level and the actual Landsat TM/ETM+ overlapped region. Our results on the discussed indices and sensors show as following: (1) Identifiable angle effects exist with a more elevated influence than that introduced by band difference among sensors; (2) The absolute difference between forward and backward direction can reach up to −0.03 to 0.1 within bands of the TM/ETM+ overlapped region; (3) The investigation at ground level indicates that there are different variations of angle effect transmitted to each remote sensing index. Regarding cases of crop canopy at various growth phases, most of the discussed indices have more than a 20% relative difference to nadir value except Normalized Difference Vegetation Index (NDVI) and Normalized Difference Water Index (NDWI) with the magnitude lower than 10%, and less than 16% of Normalized Burn Ratio (NBR). For the case of wax maturity stage, the relative difference to nadir value of Enhanced Vegetation Index (EVI), Soil-Adjusted Vegetation Index (SAVI), Ratio Vegetation Index (RVI), Char Soil Index (CSI), NBR, Normalized Difference Moisture Index (NDMI), and SWIR2/NIR exceeded 50%, while the values for NBR and NDMI can reach up to 115.8% and 206.7%, respectively; (4) Various schemes of index construction imply different propagation of angle effect uncertainty. The “difference” indices can partially suppress the directional influence, while the “ratio” indices show high potential to amplify the angle effect. This study reveals that the angle-induced uncertainty of these indices is greater than that induced by the spectrum mismatch among sensors, especially under the case of senescence. In addition, based on this work, indices with a suppressed potential of angle effect are recommended for vegetation monitoring or information retrieval to avoid unexpected effects

Binary Electroosmotic-Pump Nanoflow Gradient Generator for Miniaturized High-Performance Liquid Chromatography

High-performance liquid chromatography (HPLC) plays an important role in biotechnology, and a majority of chromatographic separations use gradient elution. While gradient generators can be built in different formats, binary pumps or quaternary pumps are most frequently used for gradient generator constructions. We have recently developed a high-pressure electroosmotic pump (EOP); the pump can be manufactured at a cost of a few hundred dollars. However, it is challenging to use this pump to deliver a gradient eluent directly. In this study, we first improve the monolith preparation by applying a pressure to the monomer solution during polymerization. We assemble a binary EOP gradient generator and discuss the relationship between the gradient profile and voltage applied to the EOP. We demonstrate the feasibility of the binary EOP gradient generator for generating a smooth and reproducible nanoflow gradient. After integration of the gradient generator into a miniaturized HPLC system, we use the HPLC system for separating peptide mixtures from trypsin-digested proteins. The performance comparison between the above miniaturized HPLC system and an Agilent 1200 HPLC system exhibits comparable efficiencies, resolutions, and peak capacities

Potential Variation of Evapotranspiration Induced by Typical Vegetation Changes in Northwest China

Evapotranspiration (ET), as a key eco-hydrological parameter, plays an important role in understanding sustainable ecosystem development. Each plant category has a unique functional trait on transpiration and photosynthesis, with ET implying that water cycle and energy transformation is linked with vegetation type. Changes in surface vegetation directly alter biophysical land surface properties, hence affecting energy and ET transfer. With the rapid increase in land surface changes, there is a need to further understand and quantify the effects of vegetation change on ET, especially over the vulnerable water-cycle region in the arid and semi-arid regions of Northwest China. We adopted the GlobalLand30 land cover and MOD16A2 in 2010 and 2020 to investigate, discuss the spatio-temporal characteristics of annual and seasonal ET of cultivated land, grassland, and forests in Northwest China, and quantify the impact on vegetation changes with absolute and relative changes from different climatic subecoregions on ET. Our results show the following: (1) Forest ET was generally the highest at 688 mm, followed by cultivated land and grassland with 200–400 mm in arid climatic subecoregions. (2) Returning cultivated land to forests and cultivated land expansion potentially enhances ET by 90–110 mm/10a, with the relative rate of change increasing by 22.1% and 45.8%, respectively, away from unchanged vegetation within identical subecoregions. (3) The ET of most investigated areas gains the highest value in summer, followed by spring, autumn, and winter. This study provides reference for sustainable ecosystem development and the reasonable utilization of limited water resources in Northwest China

Thermal Stress Analysis of Solid Oxide Fuel Cell with Z-type and Serpentine-Type Channels Considering Pressure Drop

A thermo-electro-chemo-mechanical coupled 3D model was applied to simulate the performance and thermal stress of a double-sided cathode structured solid oxide fuel cell (DSC-SOFC) with two different air channel configur.ations: Z-type parallel and triple-parallel serpentine. The distribution of temperature, current density, fuel gas and thermal stress under different voltages in DCS-SOFC was illustrated, and the output power density of the cell was analyzed considering both the electrochemical power and the dissipative power caused by the pressure drop. It was found that the Z-type parallel cell gave a better performance under a low current density, while the triple-parallel serpentine cell was more efficient at a current density higher than 6330 A.m(-2). A comparison of thermal stress distributions between the two flow field designs showed a small difference in maximum 1st principle stresses under the same operational voltages. Compared to the Z-type parallel flow field, the maximum 1st principle stress in the triple-parallel serpentine was much smaller under the same current density or electrochemical power, while much larger under the same output power. (C) 2020 The Electrochemical Society ("ECS"). Published on behalf of ECS by IOP Publishing Limited

Performing Isoelectric Focusing and Simultaneous Fractionation of Proteins on A Rotary Valve Followed by Sodium Dodecyl–Polyacrylamide Gel Electrophoresis

In this technical note, we design and fabricate a novel rotary valve and demonstrate its feasibility for performing isoelectric focusing and simultaneous fractionation of proteins, followed by sodium dodecyl–polyacrylamide gel electrophoresis. The valve has two positions. In one position, the valve routes a series of capillary loops together into a single capillary tube where capillary isoelectric focusing (CIEF) is performed. By switching the valve to another position, the CIEF-resolved proteins in all capillary loops are isolated simultaneously, and samples in the loops are removed and collected in vials. After the collected samples are briefly processed, they are separated via sodium dodecyl–polyacrylamide gel electrophoresis (SDS–PAGE, the second-D separation) on either a capillary gel electrophoresis instrument or a slab-gel system. The detailed valve configuration is illustrated, and the experimental conditions and operation protocols are discussed