Vehicle routing problem with time windows and carbon emissions: a case study in logistics distribution.

Logistics and transportation industry is not only a major energy consumer, but also a major carbon emitter. Developing green logistics is the only way for the sustainable development of the logistics industry. One of the main factors of environmental pollution is caused by carbon emissions in the process of vehicle transportation, and carbon emissions of vehicle transportation are closely related to routing, road conditions, vehicle speed, and speed fluctuations. The low-carbon vehicle routing problem with high granularity time-dependent speeds, speed fluctuations, road conditions, and time windows is proposed and formally described. In order to finely evaluate the effects of vehicle speed and speed fluctuations on carbon emissions, a graph convolutional network (GCN) is used to predict the high granularity time-dependent traffic speeds. To solve this complicated low-carbon vehicle routing problem, a hybrid genetic algorithm with adaptive variable neighborhood search is proposed to obtain vehicle routing with low carbon emissions. Finally, this method is validated using a case study with the logistics and traffic data in Jingzhou, China, and also the results show the effectiveness of this proposed method.

Clonally expanded, thyrotoxic effector CD8+ T cells driven by IL-21 contribute to checkpoint inhibitor thyroiditis.

Autoimmune toxicity occurs in up to 60% of patients treated with immune checkpoint inhibitor (ICI) therapy for cancer and represents an increasing clinical challenge for expanding the use of these treatments. To date, human immunopathogenic studies of immune-related adverse events (IRAEs) have relied on sampling of circulating peripheral blood cells rather than affected tissues. Here, we directly obtained thyroid specimens from individuals with ICI-thyroiditis, one of the most common IRAEs, and compared immune infiltrates with those from individuals with spontaneous autoimmune Hashimoto's thyroiditis (HT) or no thyroid disease. Single-cell RNA sequencing revealed a dominant, clonally expanded population of thyroid-infiltrating cytotoxic CXCR6+ CD8+ T cells (effector CD8+ T cells) present in ICI-thyroiditis but not HT or healthy controls. Furthermore, we identified a crucial role for interleukin-21 (IL-21), a cytokine secreted by intrathyroidal T follicular (TFH) and T peripheral helper (TPH) cells, as a driver of these thyrotoxic effector CD8+ T cells. In the presence of IL-21, human CD8+ T cells acquired the activated effector phenotype with up-regulation of the cytotoxic molecules interferon-γ (IFN-γ) and granzyme B, increased expression of the chemokine receptor CXCR6, and thyrotoxic capacity. We validated these findings in vivo using a mouse model of IRAEs and further demonstrated that genetic deletion of IL-21 signaling protected ICI-treated mice from thyroid immune infiltration. Together, these studies reveal mechanisms and candidate therapeutic targets for individuals who develop IRAEs.

Cu-doped CaFeO3 perovskite oxide as oxygen reduction catalyst in air cathode microbial fuel cells.

Cathode electrocatalyst is quite critical to realize the application of microbial fuel cells (MFCs). Perovskite oxides have been considered as potential MFCs cathode catalysts to replace Pt/C. Herein, Cu-doped perovskite oxide with a stable porous structure and excellent conductivity was successfully prepared through a sol-gel method. Due to the incorporation of Cu, CaFe0.9Cu0.1O3 has more micropores and a larger surface area, which are more conducive to contact with oxygen. Doping Cu resulted in more Fe3+ in B-site and thus enhanced its binding capability to oxygen molecules. The data from electrochemical test demonstrated that the as-prepared catalyst has good conductivity, high stability, and excellent ORR properties. Compared with Pt/C catalyst, CaFe0.9Cu0.1O3 exhibits a lower overpotential, which had an onset potential of 0.195 V and a half-wave potential of -0.224 V, respectively. CaFe0.9Cu0.1O3 displays an outstanding four-electron pathway for ORR mechanism and demonstrates superiors corrosion resistance and stability. The MFC with CaFe0.9Cu0.1O3 has a greater maximum power density (1090 mW m-3) rather than that of Pt/C cathode (970 mW m-3). This work demonstrated CaFe0.9Cu0.1O3 is an economic and efficient cathodic catalyst for MFCs.

Predicting joint effects of multiple land consolidation strategies on ecosystem service interactions.

Land consolidation has greatly altered land use patterns and the corresponding ecosystem services (ES) in China. However, the potential consequences of different consolidation measures for the interaction of ecosystem services are still largely unknown. Here, from a simulation perspective, we predict potential separate and joint influences of three land consolidation scenarios, i.e., reclamation of abandoned mining land, intensive construction land use and their integrated scenario, on the tradeoff/ synergic relationships of food production, carbon storage, habitat quality, and water conservation. Zhaoyuan city, one of China's top 100 counties at a green transformation stage of mining industry, is taken as a case study. Our results show that land consolidation will significantly mitigate the loss of farmland and vegetation while improving use efficiency of construction land. By 2035, food production is likely to present more trade-offs with other ES, whereas carbon storage, habitat quality, and water conservation will be highly synergic with each other. In contrast, consolidation of rural/urban construction land will be more feasible to coordinate multiple ecosystem services, as well as mitigating urban expansion and rural hollowing. This work enables us to identify the optimal combinations of multiple land consolidation measures towards sustainable ecosystem management.

Mechanical Efficiency Investigation of an Ankle-Assisted Robot for Human Walking With a Backpack-Load.

The purpose of this work is to investigate the efficiency of wearable assistive devices under different load-carriage walking. We designed an experimental platform with a lightweight ankle-assisted robot. Eight subjects were tested in three experimental conditions: free walk with load (FWL), power-off with load (POFL), and power-on with load for different levels of force at a walking speed of 3.6 km/h. We recorded the metabolic expenditure and kinematics of the subjects under three levels of load-carried (10%, 20%, and 30% of body mass). We define the critical force, where at a certain load, the robot inputs a certain force to the human body, and with the assistance of this force, the positive effect of the robot on the human body exactly compensates for the negative effect. The critical forces from the fit of the assistive force and metabolic cost curves were 130 N, 160 N, and 215 N at three different load levels. The intrinsic weight of our device increases mechanical work at the ankle as the load weight rises with 2.08 J, 2.43 J, and 2.73 J for one leg during a gait cycle. With weight bearing increasing, the ratio of the mechanical work input by the robot to the mechanical work output by the weight of the device decreases (from 0.904 to 0.717 and 0.513), verifying that the walking assistance efficiency of such devices decreases as the weight rises.

A cold shock protein promotes high-temperature microbial growth through binding to diverse RNA species.

Endowing mesophilic microorganisms with high-temperature resistance is highly desirable for industrial microbial fermentation. Here, we report a cold-shock protein (CspL) that is an RNA chaperone protein from a lactate producing thermophile strain (Bacillus coagulans 2-6), which is able to recombinantly confer strong high-temperature resistance to other microorganisms. Transgenic cspL expression massively enhanced high-temperature growth of Escherichia coli (a 2.4-fold biomass increase at 45 °C) and eukaryote Saccharomyces cerevisiae (a 2.6-fold biomass increase at 36 °C). Importantly, we also found that CspL promotes growth rates at normal temperatures. Mechanistically, bio-layer interferometry characterized CspL's nucleotide-binding functions in vitro, while in vivo we used RNA-Seq and RIP-Seq to reveal CspL's global effects on mRNA accumulation and CspL's direct RNA binding targets, respectively. Thus, beyond establishing how a cold-shock protein chaperone provides high-temperature resistance, our study introduces a strategy that may facilitate industrial thermal fermentation.

Individualized Gait Generation for Rehabilitation Robots Based on Recurrent Neural Networks.

Individualized reference gait patterns for lower limb rehabilitation robots can greatly improve the effectiveness of rehabilitation. However, previous methods can only generate customized gait patterns at several fixed discrete walking speeds and generating gaits at continuously varying speeds and stride lengths remains unsolved. This work proposes an individualized gait pattern generation method based on a recurrent neural network (RNN), which is proficient in series modeling. We collected the largest gait data set of this kind, which consists of 4,425 gait patterns from 137 subjects. Using this data set, we trained an RNN to create a function mapping from body parameters and gait parameters to a gait pattern. The experimental results indicate that our model is able to generate gait patterns at continuously varying walking speeds and stride lengths while also reducing the errors in the ankle, knee, and hip measurements by 12.83%, 20.95%, and 28.25%, respectively, compared to previous state-of-the-art method.

[Progress in biodegradation of low molecular weight polycyclic aromatic hydrocarbons].

Polycyclic aromatic hydrocarbons (PAHs) cause enormous environmental hazards that threaten human health. Bacterial degradation of PAHs has been extensively studied. Bacteria enhanced their biodegradability through multiple levels of regulatory analysis and adaptive evolution to produce diverse catabolic pathways. Based on recent developments, we address here the research progress in bioremediation technology to degrade low molecular weight polycyclic aromatic hydrocarbons.

Enhancing Bioremediation Potential of Pseudomonas putida by Developing Its Acid Stress Tolerance With Glutamate Decarboxylase Dependent System and Global Regulator of Extreme Radiation Resistance.

The extensive use of acids in a variety of manufacturing industries results in the increase of discharged acidic waste stream into the environment. Such co-pollution of acids and other organic pollutants limits the biodegradation capability of neutrophilic degraders. With high-throughput genetic techniques, we aim to improve the acid tolerance of a pollutant-degrading bacterium, Pseudomonas putida S16 by genetically engineering it with the glutamate decarboxylase (GAD)-dependent system and the global regulator (IrrE) of extreme radiation resistance. The engineered strains holding either GAD system or irrE regulator could grow under pH 4.5, compared to the wild type. They could also degrade over 90% of a selected pollutant (benzoate or nicotine) under pH 5.0 in 48 h, while no biodegradation was detected with the wild type under the same conditions. We conclude that acid stress tolerance by the possession of the GAD system or IrrE regulator in pollutant-degrading bacteria would be a promising approach to enhance their viability and biodegrading activities in bioremediation of acidic wastes.

Characterization of a Dibenzofuran-degrading strain of Pseudomonas aeruginosa, FA-HZ1.

Dibenzofuran (DBF) derivatives have caused serious environmental problems, especially those produced by paper pulp bleaching and incineration processes. Prominent for its resilient mutagenicity and toxicity, DBF poses a major challenge to human health. In the present study, a new strain of Pseudomonas aeruginosa, FA-HZ1, with high DBF-degrading activity was isolated and identified. The determined optimum conditions for cell growth of strain FA-HZ1 were a temperature of 30 °C, pH 5.0, rotation rate of 200 rpm and 0.1 mM DBF as a carbon source. The biochemical and physiological features as well as usage of different carbon sources by FA-HZ1 were studied. The new strain was positive for arginine double hydrolase, gelatinase and citric acid, while it was negative for urease and lysine decarboxylase. It could utilize citric acid as its sole carbon source, but was negative for indole and H2S production. Intermediates of DBF 1,2-dihydroxy-1,2-dihydrodibenzofuran, 1,2-dihydroxydibenzofuran, 2-hydroxy-4-(3'-oxo-3'H-benzofuran-2'-yliden)but-2-enoic acid, 2,3-dihydroxybenzofuran, 2-oxo-2-(2'-hydrophenyl)lactic acid, and 2-hydroxy-2-(2'-hydroxyphenyl)acetic acid were detected and identified through liquid chromatography-mass analyses. FA-HZ1 metabolizes DBF by both the angular and lateral dioxygenation pathways. The genomic study identified 158 genes that were involved in the catabolism of aromatic compounds. To identify the key genes responsible for DBF degradation, a proteomic study was performed. A total of 1459 proteins were identified in strain FA-HZ1, of which 100 were up-regulated and 104 were down-regulated. A novel enzyme "HZ6359 dioxygenase", was amplified and expressed in pET-28a in E. coli BL21(DE3). The recombinant plasmid was successfully constructed, and was used for further experiments to verify its function. In addition, the strain FA-HZ1 can also degrade halogenated analogues such as 2, 8-dibromo dibenzofuran and 4-(4-bromophenyl) dibenzofuran. Undoubtedly, the isolation and characterization of new strain and the designed pathways is significant, as it could lead to the development of cost-effective and alternative remediation strategies. The degradation pathway of DBF by P. aeruginosa FA-HZ1 is a promising tool of biotechnological and environmental significance.

Coupling between d-3-phosphoglycerate dehydrogenase and d-2-hydroxyglutarate dehydrogenase drives bacterial l-serine synthesis.

l-Serine biosynthesis, a crucial metabolic process in most domains of life, is initiated by d-3-phosphoglycerate (d-3-PG) dehydrogenation, a thermodynamically unfavorable reaction catalyzed by d-3-PG dehydrogenase (SerA). d-2-Hydroxyglutarate (d-2-HG) is traditionally viewed as an abnormal metabolite associated with cancer and neurometabolic disorders. Here, we reveal that bacterial anabolism and catabolism of d-2-HG are involved in l-serine biosynthesis in Pseudomonas stutzeri A1501 and Pseudomonas aeruginosa PAO1. SerA catalyzes the stereospecific reduction of 2-ketoglutarate (2-KG) to d-2-HG, responsible for the major production of d-2-HG in vivo. SerA combines the energetically favorable reaction of d-2-HG production to overcome the thermodynamic barrier of d-3-PG dehydrogenation. We identified a bacterial d-2-HG dehydrogenase (D2HGDH), a flavin adenine dinucleotide (FAD)-dependent enzyme, that converts d-2-HG back to 2-KG. Electron transfer flavoprotein (ETF) and ETF-ubiquinone oxidoreductase (ETFQO) are also essential in d-2-HG metabolism through their capacity to transfer electrons from D2HGDH. Furthermore, while the mutant with D2HGDH deletion displayed decreased growth, the defect was rescued by adding l-serine, suggesting that the D2HGDH is functionally tied to l-serine synthesis. Substantial flux flows through d-2-HG, being produced by SerA and removed by D2HGDH, ETF, and ETFQO, maintaining d-2-HG homeostasis. Overall, our results uncover that d-2-HG-mediated coupling between SerA and D2HGDH drives bacterial l-serine synthesis.

Systematic metabolic engineering of Escherichia coli for high-yield production of fuel bio-chemical 2,3-butanediol.

The production of biofuels by recombinant Escherichia coli is restricted by the toxicity of the products. 2,3-Butanediol (2,3-BD), a platform and fuel bio-chemical with low toxicity to microbes, could be a promising alternative for biofuel production. However, the yield and productivity of 2,3-BD produced by recombinant E. coli strains are not sufficient for industrial scale fermentation. In this work, the production of 2,3-BD by recombinant E. coli strains was optimized by applying a systematic approach. 2,3-BD biosynthesis gene clusters were cloned from several native 2,3-BD producers, including Bacillus subtilis, Bacillus licheniformis, Klebsiella pneumoniae, Serratia marcescens, and Enterobacter cloacae, inserted into the expression vector pET28a, and compared for 2,3-BD synthesis. The recombinant strain E. coli BL21/pETPT7-EcABC, carrying the 2,3-BD pathway gene cluster from Enterobacter cloacae, showed the best ability to synthesize 2,3-BD. Thereafter, expression of the most efficient gene cluster was optimized by using different promoters, including PT7, Ptac, Pc, and Pabc. E. coli BL21/pET-RABC with Pabc as promoter was superior in 2,3-BD synthesis. On the basis of the results of biomass and extracellular metabolite profiling analyses, fermentation conditions, including pH, agitation speed, and aeration rate, were optimized for the efficient production of 2,3-BD. After fed-batch fermentation under the optimized conditions, 73.8g/L of 2,3-BD was produced by using E. coli BL21/pET-RABC within 62h. The values of both yield and productivity of 2,3-BD obtained with the optimized biological system are the highest ever achieved with an engineered E. coli strain. In addition to the 2,3-BD production, the systematic approach might also be used in the production of other important chemicals through recombinant E. coli strains.