High-Precision Microscale Particulate Matter Prediction in Diverse Environments Using a Long Short-Term Memory Neural Network and Street View Imagery.

In this study, we propose a novel long short-term memory (LSTM) neural network model that leverages color features (HSV: hue, saturation, value) extracted from street images to estimate air quality with particulate matter (PM) in four typical European environments: urban, suburban, villages, and the harbor. To evaluate its performance, we utilize concentration data for eight parameters of ambient PM (PM1.0, PM2.5, and PM10, particle number concentration, lung-deposited surface area, equivalent mass concentrations of ultraviolet PM, black carbon, and brown carbon) collected from a mobile monitoring platform during the nonheating season in downtown Augsburg, Germany, along with synchronized street view images. Experimental comparisons were conducted between the LSTM model and other deep learning models (recurrent neural network and gated recurrent unit). The results clearly demonstrate a better performance of the LSTM model compared with other statistically based models. The LSTM-HSV model achieved impressive interpretability rates above 80%, for the eight PM metrics mentioned above, indicating the expected performance of the proposed model. Moreover, the successful application of the LSTM-HSV model in other seasons of Augsburg city and various environments (suburbs, villages, and harbor cities) demonstrates its satisfactory generalization capabilities in both temporal and spatial dimensions. The successful application of the LSTM-HSV model underscores its potential as a versatile tool for the estimation of air pollution after presampling of the studied area, with broad implications for urban planning and public health initiatives.

Characterization of Six Diamide Insecticides on Ryanodine Receptor: Resistance and Species Selectivity.

Ryanodine receptor (RyR) has been used as an insecticide target to control many destructive agricultural pests. The effectiveness of these insecticides has been limited by the spread of resistance mutations identified in pest RyRs, but the detailed molecular impacts of the individual mutations on the activity of different diamide compounds have not been fully explored. We created five HEK293 cell lines stably expressing wild type rabbit RyR1, wild type Spodoptera frugiperda RyR (Sf RyR), or Sf RyR carrying different resistance mutations, including G4891E, G4891E/I4734M, and Y4867F, respectively. R-CEPIA1er, a genetically encoded fluorescent protein, was also introduced in these cell lines to report the Ca2+ concentration in the endoplasmic reticulum. We systematically characterized the activities of six commercial diamide insecticides against different RyRs using the time-lapse fluorescence assay. Among them, cyantraniliprole (CYAN) displayed the highest activity against all three resistant Sf RyRs. The good performance of CYAN was confirmed by the toxicity assay using gene-edited Drosophila expressing the mutant RyRs, in which CYAN showed the lowest LD50 value for the double resistant mutant. In addition, we compared their acitivty between mammalian and insect RyRs and found that flubendiamide has the best insect-selectivity. The mechanism of the anti-resistance property and selectivity of the compounds was proposed based on the structural models generated by homology modeling and molecular docking. Our findings provide insights into the mechanism of insect resistance and guidance for developing effective RyR agonists that can selectively target resistant pests.

Identification of novel umami peptides from yeast extract and the mechanism against T1R1/T1R3.

Yeast extract was separated by using ultrafiltration, gel filtration chromatography, and preparative high-performance liquid chromatography for analyzing the umami mechanism. 13 kinds of umami peptides were screened out from 73 kinds of peptides which were identified in yeast extract using nanoscale ultra-performance liquid chromatography-tandem mass spectrometry and virtual screening. The umami peptides were found to have a threshold range of 0.07-0.61 mM. DWTDDVEAR exhibited a strong umami taste with a pronounced enhancement effect for monosodium glutamate. Molecular docking studies revealed that specific amino acid residues in the T1R1 subunit, including Arg316, Ser401, and Asp315, played a critical role in the umami perception with these peptides. Overall, the study highlights the potential of natural flavor enhancers and provides insights into the mechanism of umami taste perception.

Ambient air particulate total lung deposited surface area (LDSA) levels in urban Europe.

This study aims to picture the phenomenology of urban ambient total lung deposited surface area (LDSA) (including head/throat (HA), tracheobronchial (TB), and alveolar (ALV) regions) based on multiple path particle dosimetry (MPPD) model during 2017-2019 period collected from urban background (UB, n = 15), traffic (TR, n = 6), suburban background (SUB, n = 4), and regional background (RB, n = 1) monitoring sites in Europe (25) and USA (1). Briefly, the spatial-temporal distribution characteristics of the deposition of LDSA, including diel, weekly, and seasonal patterns, were analyzed. Then, the relationship between LDSA and other air quality metrics at each monitoring site was investigated. The result showed that the peak concentrations of LDSA at UB and TR sites are commonly observed in the morning (06:00-8:00 UTC) and late evening (19:00-22:00 UTC), coinciding with traffic rush hours, biomass burning, and atmospheric stagnation periods. The only LDSA night-time peaks are observed on weekends. Due to the variability of emission sources and meteorology, the seasonal variability of the LDSA concentration revealed significant differences (p = 0.01) between the four seasons at all monitoring sites. Meanwhile, the correlations of LDSA with other pollutant metrics suggested that Aitken and accumulation mode particles play a significant role in the total LDSA concentration. The results also indicated that the main proportion of total LDSA is attributed to the ALV fraction (50 %), followed by the TB (34 %) and HA (16 %). Overall, this study provides valuable information of LDSA as a predictor in epidemiological studies and for the first time presenting total LDSA in a variety of European urban environments.

Development of an efficient insecticide substrate and inhibitor screening system of insect P450s using fission yeast.

Metabolic resistance is one of the most frequent mechanisms of insecticide resistance, characterized by an increased expression of several important enzymes and transporters, especially cytochrome P450s (CYPs). Due to the large number of P450s in pests, determining the precise relationship between these enzymes and the insecticide substrates is a challenge. Herein, we developed a luminescence-based screening system for efficient identification of insecticide substrates and insect P450 inhibitors. We recombinantly expressed Bemisia tabaci CYP6CM1vQ (Bt CYP6CM1vQ) in the fission yeast Schizosaccharomyces pombe and subsequently permeabilized the yeast cells to convert them into "enzyme bags". We exploited these enzyme bags to screen the activity of twelve luciferin substrates and identified Luciferin-FEE as the optimal competing probe that was further used to characterize the metabolism of eight candidate commercial insecticides. Among them, Bt CYP6CM1vQ exhibited notable activity against pymetrozine and imidacloprid. Their binding modes were predicted by homology modeling and molecular docking, revealing the mechanisms of the metabolism. We also tested the inhibitory effect of eight known P450 inhibitors using our system and identified letrozole and 1-benzylimidazole as showing significant activity against Bt CYP6CM1vQ, with IC50 values of 23.74 µM and 1.30 µM, respectively. Their potential to be developed as an insecticide synergist was further proven by an in vitro toxicity assay using imidacloprid-resistant Bemisia tabaci. Overall, our luciferin-based enzyme bag method is capable of providing a robust and efficient screening of insect P450 substrates and, more importantly, inhibitors to overcome the resistance.

Expression and purification of snustorr snarlik protein from Plutella xylostella.

Snustorr snarlik (Snsl) is a type of extracellular protein essential for insect cuticle formation and insect survival, but is absent in mammals, making it a potential selective target for pest control. Here, we successfully expressed and purified the Snsl protein of Plutella xylostella in Escherichia coli. Two truncated forms of Snsl protein, Snsl 16-119 and Snsl 16-159, were expressed as a maltose-binding protein (MBP) fusion protein and purified to a purity above 90% after a five-step purification protocol. Snsl 16-119, forming stable monomer in solution, was crystallized, and the crystal was diffracted to a resolution of ∼10 Å. Snsl 16-159, forming an equilibrium between monomer and octamer in solution, was shown to form rod-shaped particles on negative staining electron-microscopy images. Our results lay a foundation for the determination of the structure of Snsl, which would improve our understanding of the molecular mechanism of cuticle formation and related pesticide resistance and provide a template for structure-based insecticide design.

Levels and drivers of urban black carbon and health risk assessment during pre- and COVID19 lockdown in Augsburg, Germany.

This study aimed to evaluate the levels and phenomenology of equivalent black carbon (eBC) at the city center of Augsburg, Germany (01/2018 to 12/2020). Furthermore, the potential health risk of eBC based on equivalent numbers of passively smoked cigarettes (PSC) was also evaluated, with special emphasis on the impact caused by the COVID19 lockdown restriction measures. As it could be expected, peak concentrations of eBC were commonly recorded in morning (06:00-8:00 LT) and night (19:00-22:00 LT) in all seasons, coinciding with traffic rush hours and atmospheric stagnation. The variability of eBC was highly influenced by diurnal variations in traffic and meteorology (air temperature (T), mixing-layer height (MLH), wind speed (WS)) across days and seasons. Furthermore, a marked "weekend effect" was evidenced, with an average eBC decrease of ∼35% due to lower traffic flow. During the COVID19 lockdown period, an average ∼60% reduction of the traffic flow resulted in ∼30% eBC decrease, as the health risks of eBC exposure was markedly reduced during this period. The implementation of a multilinear regression analysis allowed to explain for 53% of the variability in measured eBC, indicating that the several factors (e.g., traffic and meteorology) may contribute simultaneously to this proportion. Overall, this study will provide valuable input to the policy makers to mitigate eBC pollutant and its adverse effect on environment and human health.

Calmodulin Modulation of Insect Ryanodine Receptors.

Ryanodine receptor (RyR) is a giant calcium release channel located on the membrane of the endoplasmic reticulum (ER). Here, we report the regulation of RyRs from two major agricultural pests, diamondback moth and fall armyworm, by insect calmodulin (CaM). The recombinantly expressed full-length insect RyR could be pulled down by insect CaM in the presence of Ca2+, but the efficiency is lower compared to rabbit RyR1 and insect RyR with the CaM-binding domain (CaMBD) replaced by rabbit RyR1 sequence. Interestingly, the enhanced binding of CaM in the mutant insect RyR resulted in an increased sensitivity to the diamide insecticide chlorantraniliprole (CHL), suggesting that this CaM-CaMBD interface could be targeted by potential synergists acting as molecular glue. The thermodynamics of the binding between insect CaM and CaMBD was characterized by isothermal titration calorimetry, and the key residues responsible for the insect-specific regulation were identified through mutagenesis studies.

Investigation of COVID-19-related lockdowns on the air pollution changes in augsburg in 2020, Germany.

The COVID-19 pandemic in Germany in 2020 brought many regulations to impede its transmission such as lockdown. Hence, in this study, we compared the annual air pollutants (CO, NO, NO2, O3, PM10, PM2.5, and BC) in Augsburg in 2020 to the record data in 2010-2019. The annual air pollutants in 2020 were significantly (p < 0.001) lower than that in 2010-2019 except O3, which was significantly (p = 0.02) higher than that in 2010-2019. In a depth perspective, we explored how lockdown impacted air pollutants in Augsburg. We simulated air pollutants based on the meteorological data, traffic density, and weekday and weekend/holiday by using four different models (i.e. Random Forest, K-nearest Neighbors, Linear Regression, and Lasso Regression). According to the best fitting effects, Random Forest was used to predict air pollutants during two lockdown periods (16/03/2020-19/04/2020, 1st lockdown and 02/11/2020-31/12/2020, 2nd lockdown) to explore how lockdown measures impacted air pollutants. Compared to the predicted values, the measured CO, NO2, and BC significantly reduced 18.21%, 21.75%, and 48.92% in the 1st lockdown as well as 7.67%, 32.28%, and 79.08% in the 2nd lockdown. It could be owing to the reduction of traffic and industrial activities. O3 significantly increased 15.62% in the 1st lockdown but decreased 40.39% in the 2nd lockdown, which may have relations with the fluctuations the NO titration effect and photochemistry effect. PM10 and PM2.5 were significantly increased 18.23% an 10.06% in the 1st lockdown but reduced 34.37% and 30.62% in the 2nd lockdown, which could be owing to their complex generation mechanisms.

Effects of amino acid composition of yeast extract on the microbiota and aroma quality of fermented soy sauce.

Yeast extracts, of which amino acids are the main component, can be directly applied to improve the flavor of final soy sauce. In this study, the potential of commercial yeast extracts was explored from amino acid approach to enhance the flavor quality of soy sauce by shaping the core fermentation microbiota. Alkaline and neutral amino acids favored the competitive benefits of flavor-producing bacteria, while acidic amino acids promoted the stress resistance of the fermentation microbiota, especially the abundance of Lactobacillus, which increased to 18.03-23.78% and became the predominant microbiota. The mass ratio of neutral-nonpolar: neutral-polar: acidic: alkaline amino acids was 40: 18: 27: 15, which provided the optimal improvement of soy sauce aroma. The formulation and activated the metabolic pathways of 3-methyl-1-butyraldehyde, 3-methyl-1-butanol and 2-methyl-1-propanol through Leu and Ile, resulting in a 52.6% increase in malt-like aroma. This study provides a new idea for the regulation of soy sauce fermentation.

Structural Insight Into Ryanodine Receptor Channelopathies.

The ryanodine receptors (RyRs) are large cation-selective ligand-gated channels that are expressed in the sarcoplasmic reticulum (SR) membrane. They mediate the controlled release of Ca2+ from SR and play an important role in many cellular processes. The mutations in RyRs are associated with several skeletal muscle and cardiac conditions, including malignant hyperthermia (MH), central core disease (CCD), catecholaminergic polymorphic ventricular tachycardia (CPVT), and arrhythmogenic right ventricular dysplasia (ARVD). Recent breakthroughs in structural biology including cryo-electron microscopy (EM) and X-ray crystallography allowed the determination of a number of near-atomic structures of RyRs, including wildtype and mutant structures as well as the structures in complex with different modulating molecules. This allows us to comprehend the physiological gating and regulatory mechanisms of RyRs and the underlying pathological mechanisms of the disease-causing mutations. In this review, based on the insights gained from the available high-resolution structures of RyRs, we address several questions: 1) what are the gating mechanisms of different RyR isoforms; 2) how RyRs are regulated by multiple channel modulators, including ions, small molecules, and regulatory proteins; 3) how do disease-causing mutations affect the structure and function of RyRs; 4) how can these structural information aid in the diagnosis of the related diseases and the development of pharmacological therapies.

Recent progress in the structural study of ion channels as insecticide targets.

Ion channels, many expressed in insect neural and muscular systems, have drawn huge attention as primary targets of insecticides. With the recent technical breakthroughs in structural biology, especially in cryo-electron microscopy (cryo-EM), many new high-resolution structures of ion channel targets, apo or in complex with insecticides, have been solved, shedding light on the molecular mechanism of action of the insecticides and resistance mutations. These structures also provide accurate templates for structure-based insecticide screening and rational design. This review summarizes the recent progress in the structural studies of 5 ion channel families: the ryanodine receptor (RyR), the nicotinic acetylcholine receptor (nAChR), the voltage-gated sodium channel (VGSC), the transient receptor potential (TRP) channel, and the ligand-gated chloride channel (LGCC). We address the selectivity of the channel-targeting insecticides by examining the conservation of key coordinating residues revealed by the structures. The possible resistance mechanisms are proposed based on the locations of the identified resistance mutations on the 3D structures of the target channels and their impacts on the binding of insecticides. Finally, we discuss how to develop "green" insecticides with a novel mode of action based on these high-resolution structures to overcome the resistance.

Ryanodine Receptor as Insecticide Target.

The ryanodine receptor (RyR) is one of the primary targets of commercial insecticides. The diamide insecticide family, including flubendiamide, chlorantraniliprole, cyantraniliprole, etc., targets insect RyRs and can be used to control a wide range of destructive agricultural pests. The diamide insecticides are highly selective against lepidopteran and coleopteran pests with relatively low toxicity for non-target species, such as mammals, fishes, and beneficial insects. However, recently mutations identified on insect RyRs have emerged and caused resistance in several major agricultural pests throughout different continents. This review paper summarizes the recent findings on the structure and function of insect RyRs as insecticide targets. Specifically, we examine the structures of RyRs from target and non-target species, which reveals the molecular basis for insecticide action and selectivity. We also examine the structural and functional changes of RyR caused by the resistance mutations. Finally, we examine the progress in RyR structure-based insecticide design and discuss how this might help the development of a new generation of green insecticides.

Effects of Human, Caprine, and Bovine Milk Fat Globules on Microbiota Adhesion and Gut Microecology.

Milk fat is an essential nutrient for infant development. The effects and mechanisms of human, caprine, and bovine milk fat globules (MFGs) on the gut microbiota were investigated in this study. Human MFGs enhance the efficacy of probiotics by inhibiting pathogen function. Akkermansia and Bifidobacterium were identified as the dominant microbiota by human MFGs. Mucin and complement inhibitory proteins in human MFGs were found to inhibit different pathogens. Caprine MFGs directly promoted the colonization of probiotics and the emergence of the biomarker Allobaculum. Mucin 1 in caprine MFGs was primarily responsible for inducing probiotic adhesion. Bovine MFGs increased the abundance of Oscillospira, which reduces the risk of obesity. Due to the enrichment of cell-cell junction proteins and the lack of mucin, the regulation of gut microecology by bovine MFGs was not readily apparent. In short, this study paves the way for the development of functional infant formula.

Dairy Processing Affects the Gut Digestion and Microecology by Changing the Structure and Composition of Milk Fat Globules.

Milk fat globules (MFGs) are the major source of energy for infants' dietary intake. In this study, the effects of changes in the structure and composition of MFG after dairy processing on lipolysis and immune regulation were investigated. Pasteurized MFG tends to form protein aggregates to prevent lipolysis. However, the aggregate is rich in neutrophil degranulation products, which are effective in killing pathogens. Homogenized MFG has the lowest hydrolysis rate due to the reconstituted anti-lipase barrier and exposed apolipoprotein. Simultaneously, the reconstituted barrier can compensate for the lack of the complement cascade. Spray-dried MFG had the highest hydrolysis rate attributable to the disrupted MFG barrier and the release of lipoprotein lipase and endothelial lipase. The immunomodulatory properties of spray-dried MFG proteins are mainly mediated by the toll-like receptor (TLR) signaling pathway. This research provides the improvement basis of dairy processing and functional infant formulas.

Removal performance and mechanisms of toxic hexavalent chromium (Cr(VI)) with ZnCl2 enhanced acidic vinegar residue biochar.

Acidic vinegar residue (VR) and toxic hexavalent chromium (Cr(VI)) are unfavorable substances due to their toxicity against the environment. In this study, modified biochar was prepared to investigate the removal mechanisms of Cr(VI). The results showed that ZnCl2 could yield highly aromatic products with improved pore structures. The adsorption capacity of modified biochar reached the highest efficiency (236.81 mg g-1) when the mass ratio of ZnCl2/VR was 1, which is higher than the control (9.96 mg g-1). In addition, Cr(VI) adsorption coexisted with physical and chemical adsorption. The mechanisms of modified biochar to Cr(VI) removal included electrostatic attraction, pore filing, reduction and surface complexation. Notably, as a fermented product, VR biochar was a nitrogen-rich product; the formation of the amino group could provide a direct solid site for Cr(VI) adsorption. Subsequently, amorphous silica could be converted into silanol to provide additional adsorption sites. This work establishes the theoretical basis for efficient Cr(VI) removal and VR reuse.

Air pollution in Germany: Spatio-temporal variations and their driving factors based on continuous data from 2008 to 2018.

This study analyzed long-term observational data of particulate matter (PM2.5, PM10) variability, gaseous pollutants (CO, NO2, NOX, SO2, and O3), and meteorological factors in 412 fixed monitoring stations from January 2008 to December 2018 in Germany. Based on Hurst index analysis, the trend of atmospheric pollutants in Germany was stable during the research period. The relative correlations of gaseous pollutants and meteorological factors on PM2.5 and PM10 concentrations were analyzed by Back Propagation Neural Network model, showing that CO and temperature had the greater correlations with PM2.5 and PM10. Following that, PM2.5 and PM10 show a strong positive correlation (R2 = 0.96, p < 0.01), suggesting that the reduction of PM2.5 is essential for reducing PM pollution and enhancing air quality in Germany. Based on typical PM10/CO ratios obtained under ideal weather conditions, it is conducive to roughly estimate the contribution of natural sources. In winter, the earth's crust contributed about 20.1% to PM10. Taken together, exploring the prediction methods and analyzing the characteristic variation of pollutants will contribute an essential implication for air quality control in Germany.

Determination of microbial diversities and aroma characteristics of Beitang shrimp paste.

Beitang shrimp paste (BSP) is fermented by different parts of shrimp, such as the head (H), meat (M), or the whole shrimp (S and W). Microbial communities of BSP were dominated by Firmicutes and Proteobacteria at the phyla level and Tetragenococcus at the genus level. However, the microbial diversity of M was the lowest than the others. Non-dominant bacterial communities were presented by a mutual symbiotic model in BSP fermentation. Tetragenococcus, Halanaerobium, Streptococcus, and Brevundimonas were positively correlated with the biosynthesis of amino acids, fatty acids, and metabolic cofactors; Marinilactibacillus and Pseudomonas might be the main contributors to inorganic sulfides, nitrogen oxides, and long-chain alkanes in BSP; Psychrobacter was closely related to the ester characteristics of methyl palmitoleate and methyl hexadecanoate in H. Halanaerobium and Streptococcus promoted the production of pyrazines in S. Tetragenococcus was positively correlated with acetic acid, decanoic acid, and palmitic acid that improved the sour aroma of M. The relationship between bacteria and aroma formation under different raw materials was expected to improve the quality of BSP.

Characterization of aldehydes and hydroxy acids as the main contribution to the traditional Chinese rose vinegar by flavor and taste analyses.

The volatile aroma compounds of traditional Chinese rose vinegar were identified by headspace solid-phase micro extraction gas chromatography-mass spectrometry (HS-SPME-GC-MS) and GC-MS-olfactometry (GC-MS-O), and the metabolites were identified by silylation-GC-MS in this study. A total of 48 and 76 kinds of flavors and metabolites, respectively were detected in this study. Quantitative analysis showed that aldehydes and acids were present in relatively high amounts. Furthermore, the data colleted by the calculated odor activity values (OAVs) suggested that aldehydes are likely to contribute greatly to the aroma of traditional Chinese rose vinegar, especially, nonanal (OAV: 133, rose), 3-methyl-butanal (OAV: 57, apple-like), decanal (OAV: 23, orange peel), heptanal (OAV: 17, fruity), and dodecanal (OAV: 4-9, violet scents). Moreover, among the detected nonvolatile acids, 14 kinds of hydroxy acids, such as lactic acid, citric acid, 3-phenyllactic acid (PLA) and d-gluconic acid were detected in rose vinegar. The acids provide a well buffer system, not only greatly reduce the irritation of acetic acid, but also improve the flavor of rose vinegar. This study suggests that the fragrance and sour notes in rose vinegar are from aldehydes and hydroxy acids.

Chemical Characteristics of Three Kinds of Japanese Soy Sauce Based on Electronic Senses and GC-MS Analyses.

Japanese soy sauce has become more acceptable by Chinese consumers due to its umami taste. However, the volatile flavor compounds and taste characters have not been fully clarified. This study aimed to explore the flavor characteristics of three kinds of Japanese soy sauce, including Koikuchi Shoyu, Usukuchi Shoyu, and Amakuchi Shoyu. The secret of volatile flavor substances was investigated by Gas Chromatography-Mass Spectrometry (GC-MS) and electronic nose, while taste compounds were investigated by silylation GC-MS and electronic tongue (E-tongue). A total of 173 volatile flavor substances and 160 taste compounds were identified. In addition, 28 aroma compounds with odor activity values (OAV) ≥ 1 were considered as the typical flavors. We found that alcohols and aldehydes were in high abundance in Japanese soy sauce, but only a small portion of pyrazines and esters were. Based on electronic nose and GC-MS analysis, Koikuchi Shoyu gives more contribution to aroma compounds, while Usukuchi Shoyu and Amakuchi Shoyu give the sourness and sweetness features based on E-tongue and silylation GC-MS analysis. In this study, 50 kinds of sugars were detected that contributed to the sweetness of soy sauce. This study will provide new insight into the flavor characteristics of Japanese soy sauce that potentially contribute to the innovation and development of soy sauce.