Nitrite accumulation, denitrification kinetic and microbial evolution in the partial denitrification process: The combined effects of carbon source and nitrate concentration.

The combined effects of carbon source (HAc, HPr, Glu, Glu + HAc) and nitrate concentration (40, 80 mg/L labeling as R40, R80) on partial denitrification (PD) were discussed at C/N ratio of 2.5 (COD = 100, 200 mg/L). The optimal NO2--N and NTR reached to 67.03 mg/L, 99.14% in HAc-R80 system, and denitrification kinetics revealed the same conclusion, corresponding to higher COD utilization rate (CUR: 58.46 mgCOD/(gVSS·h)), nitrate reduction rate (NaRR: 29.94 mgN/(gVSS·h)) and nitrite accumulation rate (NiAR: 29.68 mgN/(gVSS·h)). The preference order was HAc > HPr > Glu + HAc > Glu in both R40 and R80 systems due to different metabolic pathways, however, the NO2--N accumulation and kinetic parameters of R80 group were dramatically higher than those in R40 for the same carbon source. The R80 group facilitated more concentrated biodiversity (607-808 OTUs) with Terrimonas and norank_f_Saprospiraceae responsible for high NO2--N accumulation in HAc and HPr served systems, while norank_f_norank_o_Saccharimonadales and OLB13 dominated the Glu containing systems.

Impact of Surface Chemistry of Ultrasmall Superparamagnetic Iron Oxide Nanoparticles on Protein Corona Formation and Endothelial Cell Uptake, Toxicity, and Barrier Function.

Ultrasmall superparamagnetic iron oxide nanoparticles (USPIONs) have been investigated for biomedical applications, including novel contrast agents, magnetic tracers for tumor imaging, targeted drug delivery vehicles, and magneto-mechanical actuators for hyperthermia and thrombolysis. Despite significant progress, recent clinical reports have raised concerns regarding USPION safety related to endothelial cell dysfunction; however, there is limited information on factors contributing to these clinical responses. The influence of USPION surface chemistry on nanoparticle interactions with proteins may impact endothelial cell function leading to adverse responses. Therefore, the goal of this study was to assess the effects of carboxyl-functionalized USPION (CU) or amine-functionalized USPION (AU) (approximately 30 nm diameter) on biological responses in human coronary artery endothelial cells. Increased protein adsorption was observed for AU compared with CU after exposure to serum proteins. Exposure to CU, but not AU, resulted in a concentration-dependent decrease in cell viability and perinuclear accumulation inside cytoplasmic vesicles. Internalization of CU was correlated with endothelial cell functional changes under non-cytotoxic conditions, as evidenced by a marked decreased expression of endothelial-specific adhesion proteins (eg, vascular endothelial-cadherin and platelet endothelial cell adhesion molecule-1) and increased endothelial permeability. Evaluation of downstream signaling indicated endothelial permeability is associated with actin cytoskeleton remodeling, possibly elicited by intracellular events involving reactive oxygen species, calcium ions, and the nanoparticle cellular uptake pathway. This study demonstrated that USPION surface chemistry significantly impacts protein adsorption and endothelial cell uptake, viability, and barrier function. This information will advance the current toxicological profile of USPION and improve development, safety assessment, and clinical outcomes of USPION-enabled medical products.

Bioaugmentation for low C/N ratio wastewater treatment by combining endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) in the continuous A2/O - MBBR system.

Endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) were combined in a novel A2/O - MBBR (Anaerobic Anoxic Oxic - Moving Bed Biofilm Reactor) system for low carbon/nitrogen (C/N) ratio wastewater treatment. The DPR performance was compared and the nutrient metabolism was elucidated based on the optimization of hydraulic retention time (HRT, 4-12 h) and nitrate recycling (R, 200%-600%). In the continuous-flow, the nitrate (NO3-) denitrification accompanied by nitrite (NO2-, via EPD) accumulation with the nitrate-to-nitrite transformation ratio (NTR) of 35.87%-43.31% in the anoxic zones. At HRT of 12 h with R of 500%, batch test initially revealed the DPR mechanism using both NO3- and NO2- as electron acceptor, where denitrifying phosphorus accumulation organisms (DPAOs) and denitrifying glycogen accumulation organisms (DGAOs) were the main contributors for EPD with incomplete denitrification (NO3- → NO2-). Furthermore, stoichiometry-based functional bacteria analysis displayed that higher bioactivity of DPAOs (NO2-→N2, 57.30%; NO3-→N2, 35.85%) over DGAOs (NO3-→N2, 6.85%) facilitated the anoxic NO3- reduction. Microbial community analysis suggested that Cluster I of Defluviicoccus-GAO group (∼4%) was responsible for stable NO2- accumulation performance via EPD, while increased Accumulibacter-PAO group (by ∼15%) contributed to the advanced nutrient removal. Based on the achievement of NO2- accumulation, the application feasibility of integrated EPD - DPR - Anammox for deep-level nutrient removal was discussed.

A comprehensive survey on smart contract construction and execution: paradigms, tools, and systems.

Smart contracts are regarded as one of the most promising and appealing notions in blockchain technology. Their self-enforcing and event-driven features make some online activities possible without a trusted third party. Nevertheless, problems such as miscellaneous attacks, privacy leakage, and low processing rates prevent them from being widely applied. Various schemes and tools have been proposed to facilitate the construction and execution of secure smart contracts. However, a comprehensive survey for these proposals is absent, hindering new researchers and developers from a quick start. This paper surveys the literature and online resources on smart contract construction and execution over the period 2008-2020. We divide the studies into three categories: (1) design paradigms that give examples and patterns on contract construction, (2) design tools that facilitate the development of secure smart contracts, and (3) extensions and alternatives that improve the privacy or efficiency of the system. We start by grouping the relevant construction schemes into the first two categories. We then review the execution mechanisms in the last category and further divide the state-of-the-art solutions into three classes: private contracts with extra tools, off-chain channels, and extensions on core functionalities. Finally, we summarize several challenges and identify future research directions toward developing secure, privacy-preserving, and efficient smart contracts.

Nitrite accumulation and microbial behavior by seeding denitrifying phosphorus removal sludge for partial denitrification (PD): The effect of COD/NO3- ratio.

Effect of influent COD/NO3- ratios (2.0-5.0) on partial denitrification (PD, NO3- to NO2-) was investigated by seeding denitrifying phosphorus removal (DPR) sludge at ambient temperature (16 ± 2℃). At COD/NO3- ratio of 2.5, the optimal NO2- effluent and nitrate-to-nitrite transformation ratio (NTR) reached up to 27.18 mg/L and 82.18%, respectively, and nitrate removal efficiency (NRE) (59.34 â†’ 97.98%) showed positive relationship with increasing COD/NO3- ratios. The variations were further illustrated by denitrification kinetics, where faster COD degradation (1.55 g COD/(gVSS·d)), more NO3- consumption (0.55 gN/(gVSS·d)) and higher NO2- production (0.52 gN/(gVSS·d)) were observed although the NO2- peaks happened at anoxic 30 min. Microbial analysis showed lower community diversity and more concentrated composition with dominated genera Thauera (14.10%), Terrimonas (9.40%), Saprospiraceae (13.50%) and Flavobacterium (28.23%) at COD/NO3- ratio of 2.5. Based on the achievement of PD, the application feasibility of integrated PD-DPR-Anammox in a two-sludge DPR system for deep-level nutrient removal was discussed.

Human in vitro vascularized micro-organ and micro-tumor models are reproducible organ-on-a-chip platforms for studies of anticancer drugs.

Angiogenesis is a complex process that is required for development and tissue regeneration and it may be affected by many pathological conditions. Chemicals and drugs can impact formation and maintenance of the vascular networks; these effects may be both desirable (e.g., anti-cancer drugs) or unwanted (e.g., side effects of drugs). A number of in vivo and in vitro models exist for studies of angiogenesis and endothelial cell function, including organ-on-a-chip microphysiological systems. An arrayed organ-on-a-chip platform on a 96-well plate footprint that incorporates perfused microvessels, with and without tumors, was recently developed and it was shown that survival of the surrounding tissue was dependent on delivery of nutrients through the vessels. Here we describe a technology transfer of this complex microphysiological model between laboratories and demonstrate that reproducibility and robustness of these tissue chip-enabled experiments depend primarily on the source of the endothelial cells. The model was highly reproducible between laboratories and was used to demonstrate the advantages of the perfusable vascular networks for drug safety evaluation. As a proof-of-concept, we tested Fluorouracil (1-1,000 µM), Vincristine (1-1,000 nM), and Sorafenib (0.1-100 µM), in the perfusable and non-perfusable micro-organs, and in a colon cancer-containing micro-tumor model. Tissue chip experiments were compared to the traditional monolayer cultures of endothelial or tumor cells. These studies showed that human in vitro vascularized micro-organ and micro-tumor models are reproducible organ-on-a-chip platforms for studies of anticancer drugs. The data from the 3D models confirmed advantages of the physiological environment as compared to 2D cell cultures. We demonstrated how these models can be translated into practice by verifying that the endothelial cell source and passage are critical elements for establishing a perfusable model.

Rapid hazard characterization of environmental chemicals using a compendium of human cell lines from different organs

The lack of adequate toxicity data for the vast majority of chemicals in the environment has spurred the development of new approach methodologies (NAMs). This study aimed to develop a practical high-throughput in vitro model for rapidly evaluating potential hazards of chemicals using a small number of human cells. Forty-two compounds were tested using human induced pluripotent stem cell (iPSC)-derived cells (hepatocytes, neurons, cardiomyocytes and endothelial cells), and a primary endothelial cell line. Both functional and cytotoxicity endpoints were evaluated using high-content imaging. Concentration-response was used to derive points-of-departure (POD). PODs were integrated with ToxPi and used as surrogate NAM-based PODs for risk characterization. We found chemical class-specific similarity among the chemicals tested; metal salts exhibited the highest overall bioactivity. We also observed cell type-specific patterns among classes of chemicals, indicating the ability of the proposed in vitro model to recognize effects on different cell types. Compared to available NAM datasets, such as ToxCast/Tox21 and chemical structure-based descriptors, we found that the data from the five-cell-type model was as good or even better in assigning compounds to chemical classes. Additionally, the PODs from this model performed well as a conservative surrogate for regulatory in vivo PODs and were less likely to underestimate in vivo potency and potential risk compared to other NAM-based PODs. In summary, we demonstrate the potential of this in vitro screening model to inform rapid risk-based decision-making through ranking, clustering, and assessment of both hazard and risks of diverse environmental chemicals.

Elucidating sludge characteristic, substrate transformation and microbial evolution in a two-sludge denitrifying phosphorus removal system under the impact of HRT.

Granule formation has been recognized as a promising biotechnology in denitrifying phosphorus removal (DPR) systems by facilitating phosphorus accumulation organisms (PAOs) especially denitrifying PAOs (DPAOs), and hydraulic selection made this a more difficult task in continuous operation. This study aimed at exploring the microscopic mechanism and putting forward an effective strategy for DPR granulation under the impact of hydraulic retention time (HRT) (12 h, 10 h, 8 h) in a novel Anaerobic Anoxic Oxic - Moving Bed Biofilm Reactor (A2/O - MBBR) system. With the reduction of intracellular carbon storage (CODintra) efficiency (88.58%-78.53%), nitrogen (N) (85.45%-79.11%) and phosphorus (P) (96.55%-92.47%) removals both dropped, but it exhibited a growth of anoxic phosphorus uptake rate (PURA) (3.79-5.68 mg P/(gMLVSS·h)). The batch tests associating with substrate transformation of poly-ß-hydroxyalkanoates (PHA), glycogen (Gly) agreed well with the corresponding stoichiometry of phosphorus release rate (PRR) (4.83-7.53 mg P/(gMLVSS·h)), PURA (3.55-5.43 mg P/(gMLVSS·h)), oxic phosphorus uptake rate (PURO) (6.08-6.21 mg P/(gMLVSS·h)), and DPAOs/PAOs ratios (57.17%-89.31%), indicating a shift of microbial community. DPR granules gradually stabilized with low sludge volume index (SVI5/SVI30 ratio = 1.1-1.2), dense and compact structure, higher P content (11.63%), more extracted extracellular polymeric substances (EPS) (111.40-160.31 mg/gMLVSS) as proteins/polysaccharides (PN/PS) ratios (1.70-3.47) increased, leading to better sludge settleability and cell hydrophobicity. Fluorescence in situ hybridization (FISH) results showed that PAOs (mainly Cluster I: 20.20%) were the dominant bacteria in the A2/O reactor although a small amount of Defluviicoccus (3.18-3.48%) was responsible for nitrite accumulation, while ammonium-oxidizing bacteria (AOB) (mainly Nitrosomonas: 10.75%) and nitrite-oxidizing bacteria (NOB) (mainly Nitrospira: 15.06%) were enriched in the MBBR.

A preliminary survey of Zika virus infection by nucleic acid test in the volunteer blood donor samples in Shenzhen China.

Although Zika virus (ZIKV) infection is mainly transmitted through mosquito bite, it can also be transmitted through blood transfusion. More than 500 000 cases of ZIKV infection were reported in the Americas from 2015 to 2016. Up till now, over 10 cases of imported ZIKV infection have been reported due to frequent international exchanges in the Shenzhen city of Guangdong Province, China. Unfortunately, there were no data on ZIKV infection in Chinese blood donors because it has not been included in routine screening for volunteer blood donors. As such, we performed a preliminary survey of the prevalence of ZIKV infection among volunteer blood donors in Shenzhen, China, to assess the potential risk of ZIKV infection through transfusion. A total of 9626 blood donor samples were collected and ZIKA RNA was detected by transcription-mediated amplification (TMA) nucleic acid amplification method with the Panther nucleic acid automatic analysis system of Grifols, Spain, including Procleix ZIKV Assay reagent. All the experiments in this study were conducted in accordance with the standard operating procedure of the blood center. Of the 9626 donor blood samples tested, none of these samples was Zika RNA reactive. There was no positive case from ZIKV RNA screening in this preliminary survey. There was no ZIKV presence in blood donors in Shenzhen, China, from this preliminary survey. The potential risk of ZIKV infection by transfusion is low in Shenzhen at this moment. Therefore, there is no need to add ZIKV nucleic acid test as a routine screening for blood donors.

Tissue-Engineered Bone Tumor as a Reproducible Human in Vitro Model for Studies of Anticancer Drugs.

Studies of anticancer therapies in traditional cell culture models can demonstrate efficacy of direct-acting compounds but lack the 3-dimensional arrangement of the tumor cells and their tissue-specific microenvironments, both of which are important modulators of treatment effects in vivo. Bone cells reside in complex environments that regulate their fate and function. A bioengineered human bone-tumor model has been shown to provide a microphysiological niche for studies of cancer cell behavior. Here, we demonstrate successful transfer between 2 laboratories and utility of this model in efficacy studies using well-established chemotherapeutic agents. The bioengineered human bone-tumor model consisted of Ewing sarcoma (RD-ES) cancer cell aggregates infused into tissue-engineered bone that was grown from human mesenchymal stem cell-derived differentiated into osteoblasts within mineralized bone scaffolds. The tumor model was maintained in culture for over 5 weeks and subjected to clinically relevant doses of linsitinib, doxorubicin, cisplatin, methotrexate, vincristine, dexamethasone, or MAP (methotrexate, doxorubicin, and cisplatin combination). Drug administration cycles were designed to mimic clinical treatment regimens. The bioengineered tumors were evaluated days to weeks after the cessation of treatment to monitor the potential for relapse, using bioengineered bone and ES cell monolayers as controls. Drug binding to the scaffolds and media proteins and gene expression were also evaluated. We show that a bioengineered human bone tumor can be used as a microphysiological model for preclinical studies of anticancer drugs. We found that anticancer efficacy was achieved at concentrations approximating the human Cmax, in contrast to traditional ES cell monolayers. These studies show that the bone-tumor model can be successfully transferred between laboratories and has predictive power in preclinical studies. The effects of drugs on the bone tumors and healthy bone were studied in parallel, in support of the utility of this model for identification of new therapeutic targets.

Bioaugmentation of low C/N ratio wastewater: Effect of acetate and propionate on nutrient removal, substrate transformation, and microbial community behavior.

The effect of various acetate/propionate ratios (1:0, 2:1, 1:1, 1:2, and 0:1) in a two-sludge A2/O - MBBR process was investigated. Results showed that the increased propionic/acetic ratios exerted indistinctive impact on COD (91.21-93.44%) and P (92.23-93.87%) removals, but high P content (7.42%) accelerated sludge granulation proved by SEM and EDS. Acetate favored N removal (79.52%-82.92%) with higher PURA (3.53-4.06 mgP/(gVSS·h)), while the removal declined (75.14%) due to lower PHB/PHA ratio (52.3-57.8%) with propionate as sole carbon source. Based on the stoichiometry-based quantifications, PAOs were the major contributors to nutrient removal although certain GAOs and OHO participated. The mixture ratio of 1:1 facilitated microbial diversity (995 OTUs), Rhodobacteraceae (25.63%) was responsible for high-efficient denitrifying phosphorus removal, while Defluviicoccus (15.23%) contributed to nitrite accumulation was the main competitiveness with PAOs. Nitrospira, Nitrosomonas, and Nitrosomonadaceae responsible for nitrification accounted for 7.73%, 27.11%, and 38.76% in MBBR, but the biodiversity decreased owing to the enrichment and purification.

Endothelial barrier dysfunction induced by nanoparticle exposure through actin remodeling via caveolae/raft-regulated calcium signalling.

The rapid development of modern nanotechnology has resulted in nanomaterial being use in nearly all applications of life, raising the potential risk of nanomaterial exposure alongside the need to design safe and effective materials. Previous work has demonstrated a specific effect of gold nanoparticles (GNPs) of approximately 20 nm on endothelial barrier function in vitro. To expand our understanding of this size-specific effect, titanium dioxide, silicon dioxide, and polystyrene nanoparticles (NPs) in this similar size range were studied. All tested nanoparticles were found to have minimal effects on cell viability, but exhibited a significant detrimental effect on endothelial barrier function. Nanoparticles in the size range of 20 to 30 nm were internalized by endothelial cells through caveolae/raft-mediated endocytosis, causing intracellular calcium elevation by approximately 30% at 2 hours after administration, and triggering myosin light chain kinase (MLCK)-regulated actomyosin contraction. These effects culminated in an increase in endothelial monolayer permeability across all particle types within the 20-30 nm range. This nanoparticle exposure-induced endothelial barrier dysfunction may provide valuable information for designing safer nanomaterials or potential applications of this nanoparticle exposure-induced permeability effect in biomedicine.

Surface characterization of nanoparticles using near-field light scattering.

The effect of nanoparticle surface coating characteristics on colloidal stability in solution is a critical parameter in understanding the potential applications of nanoparticles, especially in biomedicine. Here we explored the modification of the surface of poly(ethylene glycol)-coated superparamagnetic iron oxide nanoparticles (PEG-SPIOs) with the synthetic pseudotannin polygallol via interpolymer complexation (IPC). Changes in particle size and zeta potential were indirectly assessed via differences between PEG-SPIOs and IPC-SPIOs in particle velocity and scattering intensity using near-field light scattering. The local scattering intensity is correlated with the distance between the particle and waveguide, which is affected by the size of the particle (coating thickness) as well as the interactions between the particle and waveguide (related to the zeta potential of the coating). Therefore, we report here the use of near-field light scattering using nanophotonic force microscopy (using a NanoTweezerTM instrument, Halo Labs) to determine the changes that occurred in hydrated particle characteristics, which is accompanied by an analytical model. Furthermore, we found that altering the salt concentration of the suspension solution affected the velocity of particles due to the change of dielectric constant and viscosity of the solution. These findings suggest that this technique is suitable for studying particle surface changes and perhaps can be used to dynamically study reaction kinetics at the particle surface.

Silicon dioxide nanoparticle exposure affects small intestine function in an in vitro model.

The use of nanomaterials to enhance properties of food and improve delivery of orally administered drugs has become common, but the potential health effects of these ingested nanomaterials remain unknown. The goal of this study is to characterize the properties of silicon dioxide (SiO2) nanoparticles (NP) that are commonly used in food and food packaging, and to investigate the effects of physiologically realistic doses of SiO2 NP on gastrointestinal (GI) health and function. In this work, an in vitro model composed of Caco-2 and HT29-MTX co-cultures, which represent absorptive and goblet cells, was used. The model was exposed to well-characterized SiO2 NP for acute (4 h) and chronic (5 d) time periods. SiO2 NP exposure significantly affected iron (Fe), zinc (Zn), glucose, and lipid nutrient absorption. Brush border membrane intestinal alkaline phosphatase (IAP) activity was increased in response to nano-SiO2. The barrier function of the intestinal epithelium, as measured by transepithelial electrical resistance, was significantly decreased in response to chronic exposure. Gene expression and oxidative stress formation analysis showed NP altered the expression levels of nutrient transport proteins, generated reactive oxygen species, and initiated pro-inflammatory signaling. SiO2 NP exposure damaged the brush border membrane by decreasing the number of intestinal microvilli, which decreased the surface area available for nutrient absorption. SiO2 NP exposure at physiologically relevant doses ultimately caused adverse outcomes in an in vitro model.

Nanoparticle size-specific actin rearrangement and barrier dysfunction of endothelial cells.

In this work, we evaluated the impact of gold nanoparticles on endothelial cell behavior and function beyond the influence on cell viability. Five types of gold nanoparticles were studied: 5 nm and 20 nm bare gold nanoparticles, 5 nm and 20 nm gold nanoparticles with biocompatible polyethylene glycol (PEG) coating and 60 nm bare gold nanoparticles. We found that all tested gold nanoparticles did not affect cell viability significantly and reduced the reactive oxygen species (ROS) level in endothelial cells. Only 20 nm bare gold nanoparticles caused an over 50% increase in endothelial barrier permeability and slow recovery of barrier function was observed after the gold nanoparticles were removed. This impairment in endothelial barrier function was caused by unbalanced forces between intracellular tensions and paracellular forces, actin microfilament rearrangement, which occurred through a Rho/ROCK kinase-dependent pathway and broke the force balance between intracellular tensions and paracellular forces. The size-specific effect of gold nanoparticles on endothelial cells may have important implications regarding the behavior of nanoparticles in the biological system and provide valuable guidance in nanomaterial design and biomedical applications.

Isolation and Characterization of China Isolates of Duddingtonia flagrans, a Candidate of the Nematophagous Fungi for Biocontrol of Animal Parasitic Nematodes.

The nematophagous fungus Duddingtonia flagrans has been investigated as a biological agent for the control of gastrointestinal nematodes infecting domestic animals in other countries. However, D. flagrans has not been detected in China. In this study 1,135 samples were examined from 2012 to 2014; 4 D. flagrans isolates (SDH 035, SDH 091, SFH 089, SFG 170) were obtained from the feces of domestic animals and dung compost. The 4 isolates were then characterized morphologically. The SDH 035 strain was characterized by sequencing the ITS1-5.8S rDNA-ITS2 region. A BLAST search showed that the SDH 035 strain (GenBank KP257593) was 100% identical to Arthrobotrys flagrans (AF106520) and was identified as D. flagrans. The morphological plasticity of the isolated strain and the interaction of this strain with the nematode targets were observed by subjecting the infected trichostrongylide L3 to scanning electron microscopy. At 6 and 8 hr after trichostrongylide L(3) was added, hyphal ramifications were observed and L(3) were captured, respectively. Scanning electron micrographs were obtained at 0, 6, 12, 18, 24, 30, 36, 42, and 48 hr, where 0 is the time when trichostrongylide L(3) were first captured by the fungus. The details of the capture process by the fungus are also described. Chlamydospores were observed in the body of L(3) in the late stage of digestion. A sticky substance and bacteria could be observed in contact areas between predation structures and nematode cuticle.

Synthesis and anticancer evaluation of vitamin K(3) analogues.

Novel vitamin K(3) analogues were synthesized and evaluated for their anticancer activity. Compound 6, 9, 10, 11, 14, and (+/-)15 demonstrated a strong inhibitory activity against the tumor cells of A-549, Hep G2, MCF7, MES-SA, MES-SA/Dx5, MKN45, SW-480, and TW-039. Compound (+/-)15 displayed potent tumor cell cytotoxicity, and compound 14 selectively affected MCF7, even though it did not influence normal cells Detroit551 and WI-38. Compound (+/-)15 inhibited MES-SA and MES-SA/Dx5, and this specific result shows that compound (+/-)15 may become a good anticancer drug candidate.