Search

Typical crystallizable fragment (Fc) glycans attached to the CH2 domain in therapeutic monoclonal antibodies (mAbs) are core-fucosylated and asialo-biantennary complex-type glycans, e.g., G2F (full galactosylation), G1aF (terminal galactosylation on the Man α1-6 arm), G1bF (terminal galactosylation on the Man α1-3 arm), and G0F (non-galactosylation). Terminal galactose (Gal) residues of Fc-glycans are known to influence effector functions such as antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity (CDC), but the impact of the G1F isomers (G1aF and G1bF) on the effector functions has not been reported. Here, we prepared four types of glycoengineered anti-CD20 mAbs bearing homogeneous G2F, G1aF, G1bF, or G0F (G2F mAb, G1aF mAb, G1bF mAb, or G0F mAb, respectively), and evaluated their biological activities. Interestingly, G1aF mAb showed higher C1q- and FcγR-binding activities, CDC activity, and FcγR-activation property than G1bF mAb. The activities of G1aF mAb and G1bF mAb were at the same level as G2F mAb and G0F mAb, respectively. Hydrogen–deuterium exchange/mass spectrometry analysis of dynamic structures of mAbs revealed the greater involvement of the terminal Gal residue on the Man α1-6 arm in the structural stability of the CH2 domain. Considering that mAbs interact with FcγR and C1q via their hinge proximal region in the CH2 domain, the structural stabilization of the CH2 domain by the terminal Gal residue on the Man α1-6 arm of Fc-glycans may be important for the effector functions of mAbs. To our knowledge, this is the first report showing the impact of G1F isomers on the effector functions and dynamic structure of mAbs. Abbreviations: ABC, ammonium bicarbonate solution; ACN, acetonitrile; ADCC, antibody-dependent cell-mediated cytotoxicity; C1q, complement component 1q; CDC, complement-dependent cytotoxicity; CQA, critical quality attribute; Endo, endo-β-N-acetylglucosaminidase; FA, formic acid; Fc, crystallizable fragment; FcγR, Fcγ receptors; Fuc, fucose; Gal, galactose; GlcNAc, N-acetylglucosamine; GST, glutathione S-transferase; HER2, human epidermal growth factor receptor 2; HDX, hydrogen–deuterium exchange; HILIC, hydrophilic interaction liquid chromatography; HLB-SPE, hydrophilic-lipophilic balance–solid-phase extraction; HPLC, high-performance liquid chromatography; mAb, monoclonal antibody; Man, mannose; MS, mass spectrometry; PBS, phosphate-buffered saline; SGP, hen egg yolk sialylglycopeptides.

We characterized SaHEX, which is a glycoside hydrolase (GH) family 20 exo-β-N-acetylhexosaminidase found in Streptomyces avermitilis. SaHEX exolytically hydrolyzed chitin oligosaccharides from their non-reducing ends, and yielded N-acetylglucosamine (GlcNAc) as the end product. According to the initial rate of substrate hydrolysis, the rates of (GlcNAc)3 and (GlcNAc)5 hydrolysis were greater than the rates for the other oligosaccharides. The enzyme exhibited antifungal activity against Aspergillus niger, which was probably due to hydrolytic activity with regard to chitin in the hyphal tips. Therefore, SaHEX has potential for use in GlcNAc production and food preservation.

The submandibular fossa is a concavity in the lingual part of the mandibular body, inferior to the mylohyoid line, in the lower molar region, where part of the submandibular gland is situated. Around the submandibular fossa are many anatomical structures implicated in implant surgery, salivary calculus, and static bone cavity. Therefore, dentists need a thorough understanding of the surgical anatomy in the area of the submandibular fossa to preclude complications including hemorrhages, nerve injuries, ductal strictures, and perforation of the lingual cortical bone. In this chapter, the anatomy and its variations in the area of the submandibular fossa are reviewed and clinical considerations are described.

3D reconstruction models are useful for many situations in maintenance and decommissioning work at nuclear power plants (NPPs). There is a method to make the models from color and depth images. When using the method, it is necessary to scan a target environment without missing in order to make detailed and precise models. However, work sites at NPP are very complicated, and it is difficult to scan without missing. In this study, we aim to develop a scanning support system that enables users to make 3D reconstruction models without missing even in a very complicated environment such as NPPs. The system reminds and encourages users to scan work sites by visualizing unscanned area using an algorithm extended truncated signed distance function.

While chemically and thermally modified citrus pectin (MCP) has already been studied for health benefits, it is unknown how size-fractionated oligo- and polysaccharides differentially affect cancer cell behavior. We produced thermally MCP and fractionated it by molecular size to evaluate the effect these polymers have on cancer cells. MCP30/10 (between 30 and 10 kDa) had more esterified homogalacturonans (HG) and fewer rhamnogalacturonans (RG-I) than MCP and MCP30 (higher than 30 kDa), while MCP10/3 (between 10 and 3 kDa) showed higher amounts of type I arabinogalactans (AGI) and lower amounts of RG-I. MCP3 (smaller than 3 kDa) presented less esterified HG and the lowest amount of AGI and RG-I. Our data indicate that the enrichment of de-esterified HG oligomers and the AGI and RG-I depletions in MCP3, or the increase of AGI and loss of RGI in MCP30/10, enhance the anticancer behaviors by inhibiting migration, aggregation, and proliferation of cancer cells.

We summarized our recently-published papers on greenhouse gas exchanges at two important boreal regions underlain by permafrost: eastern Siberia and interior Alaska. Relevant literatures were also referred to, and future research directions on the high-latitude terrestrial processes were suggested. Long-term monitoring of CO2 fluxes at the boreal forests revealed that anomalous weather and disturbances changed the CO2 balance. More than a decade is required to return to a CO2 sink at burned forests in interior Alaska. Anomalous high precipitation altered the forest structure in eastern Siberia, shifting the overstory/understory contributions to the CO2 balance. The CH4 emissions were higher in the two boreal forests than in the other boreal forests. Upscaling the in-situ observations and comparisons with top-down approaches revealed considerable inconsistencies exist among the approaches. We recommend the following directions in future research. First, long-term monitoring is indispensable to detect the effect of climate change on ecosystems. Second, disturbance impacts, including fire, thermokarst, and wet spells, need to be quantified. Third, further observations are necessary for constraining CH4 exchange models. Finally, reconciling top-down and bottom-up approaches is required to reduce uncertainty. Prompt sharing of observed data and model products is crucial to improve our understanding of high-latitude processes.

Wetlands are the largest global natural methane (CH4) source, and emissions between 50 and 70° N latitude contribute 10–30 % to this source. Predictive capability of land models for northern wetland CH4 emissions is still low due to limited site measurements, strong spatial and temporal variability in emissions, and complex hydrological and biogeochemical dynamics. To explore this issue, we compare wetland CH4 emission predictions from the Community Land Model 4.5 (CLM4.5-BGC) with site- to regional-scale observations. A comparison of the CH4 fluxes with eddy flux data highlighted needed changes to the model's estimate of aerenchyma area, which we implemented and tested. The model modification substantially reduced biases in CH4 emissions when compared with CarbonTracker CH4 predictions. CLM4.5 CH4 emission predictions agree well with growing season (May–September) CarbonTracker Alaskan regional-level CH4 predictions and site-level observations. However, CLM4.5 underestimated CH4 emissions in the cold season (October–April). The monthly atmospheric CH4 mole fraction enhancements due to wetland emissions are also assessed using the Weather Research and Forecasting-Stochastic Time-Inverted Lagrangian Transport (WRF-STILT) model coupled with daily emissions from CLM4.5 and compared with aircraft CH4 mole fraction measurements from the Carbon in Arctic Reservoirs Vulnerability Experiment (CARVE) campaign. Both the tower and aircraft analyses confirm the underestimate of cold-season CH4 emissions by CLM4.5. The greatest uncertainties in predicting the seasonal CH4 cycle are from the wetland extent, cold-season CH4 production and CH4 transport processes. We recommend more cold-season experimental studies in high-latitude systems, which could improve the understanding and parameterization of ecosystem structure and function during this period. Predicted CH4 emissions remain uncertain, but we show here that benchmarking against observations across spatial scales can inform model structural and parameter improvements.

Understanding how high-latitude terrestrial productivity and evapotranspiration change in association with rising atmospheric CO2 concentration ([CO2]), also known as ‘CO2 fertilization’, is important for predicting future climate change. To quantify the magnitude of this fertilization effect, we have developed a big-leaf model that couples photosynthesis and stomatal conductance processes. This model was inverted by inputting eddy covariance CO2 and H2O fluxes from four black spruce forests in Alaska to infer spatially representative ecophysiological parameters using a global optimization technique. Inferred seasonal variations in a maximum carboxylation rate at 25 °C per unit leaf area and stomatal conductance suggest greater photosynthetic capacity per unit leaf area during the mid-growing season, compared to spring and autumn. The interannual variability of parameters suggest that warm summers stimulate photosynthetic capacity and dry summers force stomatal regulation. Based on the model with optimized parameters, small but clear increases in gross primary productivity (GPP) and decreases in latent heat flux (LE) were estimated to be associated with rising [CO2] from 2002 to 2014 (p

The latitudinal gradient of the start of the growing season (SOS) and the end of the growing season (EOS) were quantified in Alaska (61°N to 71°N) using satellite-based and ground-based datasets. The Alaskan evergreen needleleaf forests are sparse and the understory vegetation has a substantial impact on the satellite signal. We evaluated SOS and EOS of understory and tundra vegetation using time-lapse camera images. From the comparison of three SOS algorithms for determining SOS from two satellite datasets (SPOT-VEGETATION and Terra-MODIS), we found that the satellite-based SOS timing was consistent with the leaf emergence of the forest understory and tundra vegetation. The ensemble average of SOS over all satellite algorithms can be used as a measure of spring leaf emergence for understory and tundra vegetation. In contrast, the relationship between the ground-based and satellite-based EOSs was not as strong as that of SOS both for boreal forest and tundra sites because of the large biases between those two EOSs (19 to 26 days). The satellite-based EOS was more relevant to snowfall events than the senescence of understory or tundra. The plant canopy radiative transfer simulation suggested that 84–86% of the NDVI seasonal amplitude could be a reasonable threshold for the EOS determination. The latitudinal gradients of SOS and EOS evaluated by the satellite and ground data were consistent and the satellite-derived SOS and EOS were 3.5 to 5.7 days degree− 1 and − 2.3 to − 2.7 days degree− 1, which corresponded to the spring (May) temperature sensitivity of − 2.5 to − 3.9 days °C− 1 in SOS and the autumn (August and September) temperature sensitivity of 3.0 to 4.6 days °C− 1 in EOS. This demonstrates the possible impact of phenology in spruce forest understory and tundra ecosystems in response to climate change in the warming Artic and sub-Arctic regions.\ud \ud

Ecosystem-scale methane (CH 4 ) exchange was observed in a poorly-drained black spruce forest over permafrost in interior Alaska during the snow-free seasons of 2011–2013, using the eddy covariance technique. The magnitude of average CH 4 exchange differed depending on wind direction, reflecting spatial variation in soil moisture condition around the observation tower, due to elevation change within the small catchment. In the drier upper position, the seasonal variation in CH 4 emission was explained by the variation in soil water content only. In the wetter bottom, however, in addition to soil temperature and soil water content, seasonal thaw depth of frozen soil was also an important variable explaining the seasonal variation in CH 4 exchange for this ecosystem. Total snow-free season (day of year 134–280) CH 4 exchanges were 12.0 ± 1.0, 19.6 ± 3.0, and 36.6 ± 4.4 mmol m −2 season −1 for the drier upper position, moderately wet area, and wetter bottom of the catchment, respectively. Observed total season CH 4 emission was nearly one order smaller than those reported in other northern wetlands, due probably to the relatively low ground water level and low soil temperature. The interannual variation of total snow-free season CH 4 emission in the wetter bottom of the catchment was influenced by the amount of rainfall and thaw depth. On the other hand, in the drier upper position the amount of rainfall did not strongly affect the total season CH 4 emission. Different responses of CH 4 exchange to environmental conditions, depending on the position of a small catchment, should be considered when estimating the spatial variation in CH 4 exchange accurately in ecosystems over permafrost.

A safety assessment was conducted for microbial collagenase (COL) enzyme expressed in Streptomyces violaceoruber. The acute oral toxicity of COL was examined in Sprague-Dawley rats and the LD50 of COL via single oral administration to rats was higher than 2000 mg/kg body weight. A 13-week oral gavage study of COL showed no adverse effects due to the enzyme up to a dose of 234.9 mg total organic solids (TOS)/kg body weight per day (NOAEL). A bacterial reverse mutation test showed no mutagenic activity at the highest dose (4698 μg TOS per plate). In the mouse lymphoma TK assay, a positive result was observed at the highest dose of 4698 μg TOS/mL although it had low reproducibility. To confirm the chromosome aberration potential, an in vivo micronucleus test was conducted that demonstrated the lack of mutagenic potential on the bone marrow of rats at doses up to 1879 mg TOS/kg body weight per day. The results of the genotoxicity studies and acute and subchronic rat studies support the safe use in food production of collagenase produced from S. violaceoruber.

Free-Air CO2 Enrichment (FACE) was conceived as an experimental method to measure plant responses to elevated CO2 in natural environments rather than in chambered or controlled environments. However, due to the difficulty of controlling elevated CO2 concentrations in turbulent air, the range of fluctuations of CO2 in FACE experiments are more than 10-fold greater than plants experience in natural conditions. One early study reported that photosynthetic increases of leaves in 40- and 80-s periods of oscillating elevated CO2 were only about 68% of those in leaves exposed to constant elevated CO2 with the same mean CO2 concentration. Later whole-plant studies reported smaller increases of responses in 60-s periods of oscillating elevated CO2 compared to constant elevated CO2 with the same mean concentration. After eliminating problematic data from studies that predicted plant responses in FACE to be only 45% of responses in open top chambers, we calculated that yields increased 65% as much in fluctuating elevated CO2 of FACE as in constant elevated CO2. The smaller plant responses in fluctuating elevated CO2 can be attributed partially to the non-linear, convex-upward curved response of photosynthesis to CO2 concentration, but other unknown mechanisms must exist. Future leaf chamber studies and FACE studies should include fundamental photosynthetic physiologists who can focus on uncovering the mechanisms responsible for lower photosynthetic, biomass, and yield response in both regular waveform oscillating and irregular fluctuating elevated CO2. Because CO2 fluctuates in FACE and recent experiments indicate reduced photosynthesis and growth under fluctuating CO2, responses of plants in FACE are likely to underestimate the benefits of rising CO2. We found that a correction factor of about 1.5 needs to be applied to FACE results. While responses to elevated CO2 in FACE experiments are smaller than those in chamber experiments, FACE responses are obtained in natural conditions not available in chambers and thus are conservative regarding future projections of agricultural productivity.

For this study, a Distance Information Display System (DIDS) was developed and evaluated. Through its very simple operation, the system captures images of target objects with an RGB-D camera and automatically measures target object lengths and gaps. The system extracts and displays the lengths that workers might want to measure using a heuristic filtering process. An experiment was conducted to evaluate the DIDS and to compare the system with existing systems. The experimentally obtained results demonstrate that the DIDS can resolve problems of manual measurement and reduce the measurement time. Furthermore, results show that the DIDS is superior to existing methods in terms of ease of operation., 9th International Symposium on Symbiotic Nuclear Power Systems for 21st Century, July 9-11, 2018, Heilongjiang, China.

In response to the successful use of monoclonal antibodies (mAbs) in the treatment of various diseases, systems for expressing recombinant mAbs using transgenic animals or plants have been widely developed. The silkworm (Bombyx mori) is a highly domesticated insect that has recently been used for the production of recombinant proteins. Because of their cost-effective breeding and relatively easy production scale-up, transgenic silkworms show great promise as a novel production system for mAbs. In this study, we established a transgenic silkworm stably expressing a human-mouse chimeric anti-CD20 mAb having the same amino acid sequence as rituximab, and compared its characteristics with rituximab produced by Chinese hamster ovary (CHO) cells (MabThera®). The anti-CD20 mAb produced in the transgenic silkworm showed a similar antigen-binding property, but stronger antibody-dependent cell-mediated cytotoxicity (ADCC) and weaker complement-dependent cytotoxicity (CDC) compared to MabThera. Post-translational modification analysis was performed by peptide mapping using liquid chromatography/mass spectrometry. There was a significant difference in the N-glycosylation profile between the CHO− and the silkworm-derived mAbs, but not in other post-translational modifications including oxidation and deamidation. The mass spectra of the N-glycosylated peptide revealed that the observed biological properties were attributable to the characteristic N-glycan structures of the anti-CD20 mAbs produced in the transgenic silkworms, i.e., the lack of the core-fucose and galactose at the non-reducing terminal. These results suggest that the transgenic silkworm may be a promising expression system for the tumor-targeting mAbs with higher ADCC activity.

// Yosuke Harazono 1, 2 , Dhong Hyo Kho 1 , Vitaly Balan 3 , Kosei Nakajima 1 , Victor Hogan 1 , Avraham Raz 1 1 Departments of Oncology and Pathology, School of Medicine, Wayne State University, and Karmanos Cancer Institute, Detroit, MI 48201, USA 2 Department of Maxillofacial Surgery, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8510, Japan 3 Everon Biosciences, Buffalo, NY 14203, USA Correspondence to: Avraham Raz, e-mail: raza@karmanos.org Keywords: galectin-3, Na+/K+-ATPase, multidrug resistance, P-glycoprotein, phosphorylation Received: May 13, 2015 Accepted: June 05, 2015 Published: June 17, 2015 ABSTRACT Galectin-3 (Gal-3, LGALS3) is a pleotropic versatile, 29–35 kDa chimeric gene product, and involved in diverse physiological and pathological processes, including cell growth, homeostasis, apoptosis, pre-mRNA splicing, cell-cell and cell-matrix adhesion, cellular polarity, motility, adhesion, activation, differentiation, transformation, signaling, regulation of innate/adaptive immunity, and angiogenesis. In multiple diseases, it was found that the level of circulating Gal-3 is markedly elevated, suggesting that Gal-3-dependent function is mediated by specific interaction with yet an unknown ubiquitous cell-surface protein. Recently, we showed that Gal-3 attenuated drug-induced apoptosis, which is one of the mechanisms underlying multidrug resistance (MDR). Here, we document that MDR could be mediated by Gal-3 interaction with the house-keeping gene product e.g., Na + /K + -ATPase, and P-glycoprotein (P-gp). Gal-3 interacts with Na + /K + -ATPase and induces the phosphorylation of P-gp. We also find that Gal-3 binds P-gp and enhances its ATPase activity. Furthermore Gal-3 antagonist suppresses this interaction and results in a decrease of the phosphorylation and the ATPase activity of P-gp, leading to an increased sensitivity to doxorubicin-mediated cell death. Taken together, these findings may explain the reported roles of Gal-3 in diverse diseases and suggest that a combined therapy of inhibitors of Na + /K + -ATPase and Gal-3, and a disease specific drug(s) might be superior to a single therapeutic modality.