Integrated mRNA-miRNA transcriptome profiling of blood immune responses potentially related to pulmonary fibrosis in forest musk deer.

Background: Forest musk deer (FMD, Moschus Berezovskii) is a critically endangered species world-widely, the death of which can be caused by pulmonary disease in the farm. Pulmonary fibrosis (PF) was a huge threat to the health and survival of captive FMD. MicroRNAs (miRNAs) and messenger RNAs (mRNAs) have been involved in the regulation of immune genes and disease development. However, the regulatory profiles of mRNAs and miRNAs involved in immune regulation of FMD are unclear. Methods: In this study, mRNA-seq and miRNA-seq in blood were performed to constructed coexpression regulatory networks between PF and healthy groups of FMD. The hub immune- and apoptosis-related genes in the PF blood of FMD were explored through Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Further, protein-protein interaction (PPI) network of immune-associated and apoptosis-associated key signaling pathways were constructed based on mRNA-miRNA in the PF blood of the FMD. Immune hub DEGs and immune hub DEmiRNAs were selected for experimental verification using RT-qPCR. Results: A total of 2744 differentially expressed genes (DEGs) and 356 differentially expressed miRNAs (DEmiRNAs) were identified in the PF blood group compared to the healthy blood group. Among them, 42 DEmiRNAs were negatively correlated with 20 immune DEGs from a total of 57 correlations. The DEGs were significantly associated with pathways related to CD molecules, immune disease, immune system, cytokine receptors, T cell receptor signaling pathway, Th1 and Th2 cell differentiation, cytokine-cytokine receptor interaction, intestinal immune network for IgA production, and NOD-like receptor signaling pathway. There were 240 immune-related DEGs, in which 186 immune-related DEGs were up-regulated and 54 immune-related DEGs were down-regulated. In the protein-protein interaction (PPI) analysis of immune-related signaling pathway, TYK2, TLR2, TLR4, IL18, CSF1, CXCL13, LCK, ITGB2, PIK3CB, HCK, CD40, CD86, CCL3, CCR7, IL2RA, TLR3, and IL4R were identified as the hub immune genes. The mRNA-miRNA coregulation analysis showed that let-7d, miR-324-3p, miR-760, miR-185, miR-149, miR-149-5p, and miR-1842-5p are key miRNAs that target DEGs involved in immune disease, immune system and immunoregulation. Conclusion: The development and occurrence of PF were significantly influenced by the immune-related and apoptosis-related genes present in PF blood. mRNAs and miRNAs associated with the development and occurrence of PF in the FMD.

Overexpression of Calcineurin B-like Interacting Protein Kinase 31 Promotes Lodging and Sheath Blight Resistance in Rice.

A breakthrough "Green Revolution" in rice enhanced lodging resistance by using gibberellin-deficient semi-dwarf varieties. However, the gibberellic acid (GA) signaling regulation on rice disease resistance remains unclear. The resistance test showed that a positive GA signaling regulator DWARF1 mutant d1 was more susceptible while a negative GA signaling regulator Slender rice 1 (SLR1) mutant was less susceptible to sheath blight (ShB), one of the major rice diseases, suggesting that GA signaling positively regulates ShB resistance. To isolate the regulator, which simultaneously regulates rice lodging and ShB resistance, SLR1 interactors were isolated. Yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC), and Co-IP assay results indicate that SLR1 interacts with Calcineurin B-like-interacting protein kinase 31 (CIPK31). cipk31 mutants exhibited normal plant height, but CIPK31 OXs showed semi-dwarfism. In addition, the SLR1 level was much higher in CIPK31 OXs than in the wild-type, suggesting that CIPK31 OX might accumulate SLR1 to inhibit GA signaling and thus regulate its semi-dwarfism. Recently, we demonstrated that CIPK31 interacts and inhibits Catalase C (CatC) to accumulate ROS, which promotes rice disease resistance. Interestingly, CIPK31 interacts with Vascular Plant One Zinc Finger 2 (VOZ2) in the nucleus, and expression of CIPK31 accumulated VOZ2. Inoculation of Rhizoctonia solani AG1-IA revealed that the voz2 mutant was more susceptible to ShB. Thus, these data prove that CIPK31 promotes lodging and ShB resistance by regulating GA signaling and VOZ2 in rice. This study provides a valuable reference for rice ShB-resistant breeding.

[Study on mechanism of inhibiting proliferation of head and neck cancer cells by citral, active ingredient of lemon essential oil].

To explore the active substances exerting anti-tumour effect in lemon essential oil and the molecular mechanism inhibiting the proliferation of head and neck cancer cells SCC15 and CAL33, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay(MTT) was utilized to identify the active component inhibiting the proliferation of head and neck cancer cells, namely citral. The IC_(50) of citral inhibiting the proliferation of head and neck cancer cells and normal cells were also determined. In addition, a 5-ethynyl-2'-deoxyuridine(EdU) staining assay was used to detect the effect of citral on the proliferation rate of head and neck cancer cells, and a colony formation assay was used to detect the effect of citral on tumor sphere formation of head and neck cancer cells in vitro. The cell cycle arrest and apoptosis induction of head and neck cancer cells by citral were evaluated by flow cytometry, and Western blot was used to detect the effect of citral on the expression levels of cell cycle-and apoptosis-related proteins in head and neck cancer cells. The findings indicated that citral could effectively inhibit the proliferation and growth of head and neck cancer cells, with anti-tumor activity, and its half inhibitory concentrations for CAL33 and SCC15 were 54.78 and 25.23 µg·mL~(-1), respectively. Furthermore, citral arrested cell cycle at G_2/M phase by down-regulating cell cycle-related proteins such as S-phase kinase associated protein 2(SKP2), C-MYC, cyclin dependent kinase 1(CDK1), and cyclin B. Moreover, citral increased the cysteinyl aspartate-specific proteinase-3(caspase-3), cysteinyl aspartate-specific proteinase-9(caspase-9), and cleaved poly ADP-ribose polymerase(PARP). It up-regulated the level of autophagy-related proteins including microtubule associated protein 1 light chain 3B(LC3B), sequestosome 1(P62/SQSTM1), autophagy effector protein Beclin1(Beclin1), and lysosome-associate membrane protein 1(LAMP1), suggesting that citral could effectively trigger cell apoptosis and cell autophagy in head and neck cancer cells. Furthermore, the dual-tagged plasmid system mCherry-GFP-LC3 was used, and it was found that citral impeded the fusion of autophagosomes and lysosomes, leading to autophagic flux blockage. Collectively, our findings reveal that the main active anti-proliferation component of lemon essential oil is citral, and this component has a significant inhibitory effect on head and neck cancer cells. Its underlying molecular mechanism is that citral induces apoptosis and autophagy by cell cycle arrest and ultimately inhibits cell proliferation.

Exploration of hypoglycemic peptides from porcine collagen based on network pharmacology and molecular docking.

In recent years, the extraction of hypoglycemic peptides from food proteins has gained increasing attention. Neuropeptides, hormone peptides, antimicrobial peptides, immune peptides, antioxidant peptides, hypoglycemic peptides and antihypertensive peptides have become research hotspots. In this study, bioinformatic methods were used to screen and predict the properties of pig collagen-derived hypoglycemic peptides, and their inhibitory effects on α-glucosidase were determined in vitro. Two peptides (RL and NWYR) were found to exhibit good water solubility, adequate ADMET (absorption, distribution, metabolism, elimination, and toxicity) properties, potentially high biological activity, and non-toxic. After synthesizing these peptides, NWYR showed the best inhibitory effect on α-glucosidase with IC50 = 0.200±0.040 mg/mL, and it can regulate a variety of biological processes, play a variety of molecular functions in different cellular components, and play a hypoglycemic role by participating in diabetic cardiomyopathy and IL-17 signaling pathway. Molecular docking results showed that NWYR had the best binding effect with the core target DPP4 (4n8d), with binding energy of -8.8 kcal/mol. NWYR mainly bonded with the target protein through hydrogen bonding, and bound with various amino acid residues such as Asp-729, Gln-731, Leu-765, etc., thus affecting the role of the target in each pathway. It is the best core target for adjuvant treatment of T2DM. In short, NWYR has the potential to reduce type 2 diabetes, providing a basis for further research or food applications as well as improved utilization of pig by-products. However, in subsequent studies, it is necessary to further verify the hypoglycemic ability of porcine collagen active peptide (NWYR), and explore the hypoglycemic mechanism of NWYR from multiple perspectives such as key target genes, protein expression levels and differences in metabolites in animal models of hyperglycemia, which will provide further theoretical support for its improvement in the treatment of T2DM.

Effects of the catastrophic 2020 Yangtze River seasonal floods on microcystins and environmental conditions in Three Gorges Reservoir Area, China.

Introduction: During July and August 2020, Three Gorges Reservoir Area (TGRA) suffered from catastrophic seasonal floods. Floods changed environmental conditions and caused increase in concentration of microcystins (MCs) which is a common and potent cyanotoxin. However, the effects and seasonal variations of MCs, cyanobacteria, and environmental conditions in TGRA after the 2020 Yangtze River extreme seasonal floods remain largely unclear, and relevant studies are lacking in the literature. Methods: A total of 12 representative sampling sites were selected to perform concentration measurement of relevant water quality objectives and MCs in the representative area of the TGRA. The sampling period was from July 2020 to October 2021, which included the flood period. Organic membrane filters were used to perform the DNA extraction and analyses of the 16S rRNA microbiome sequencing data. Results: Results showed the seasonal floods result in significant increases in the mean values of microcystin-RR (MCRR), microcystin-YR (MCYR), and microcystin-LR (MCLR) concentration and some water quality objectives (i.e., turbidity) in the hinterland of TGRA compared with that in non-flood periods (p < 0.05). The mean values of some water quality objectives (i.e., total nitrogen (TN), total phosphorus (TP), total dissolved phosphorus (TDP), and turbidity), MC concentration (i.e., MCRR, MCYR, and MCLR), and cyanobacteria abundance (i.e., Cyanobium_PCC-6307 and Planktothrix_NIVA-CYA_15) displayed clear tendency of increasing in summer and autumn and decreasing in winter and spring in the hinterland of TGRA. Discussions: The results suggest that seasonal floods lead to changes in MC concentration and environmental conditions in the hinterland of TGRA. Moreover, the increase in temperature leads to changes in water quality objectives, which may cause water eutrophication. In turn, water eutrophication results in the increase in cyanobacteria abundance and MC concentration. In particular, the increased MC concentration may further contribute to adverse effects on human health.

NMDARs activation regulates endothelial ferroptosis via the PP2A-AMPK-HMGB1 axis.

N-methyl-D-aspartate receptors (NMDARs) are ligand-gated, voltage-dependent channels of the ionotropic glutamate receptor family. The present study explored whether NMDAR activation induced ferroptosis in vascular endothelial cells and its complicated mechanisms in vivo and in vitro. Various detection approaches were used to determine the ferroptosis-related cellular iron content, lipid reactive oxygen species (LOS), siRNA molecules, RNA-sequence, MDA, GSH, and western blotting. The AMPK activator Acadesine (AICAR), HMGB1 inhibitor glycyrrhizin (GLY), PP2A inhibitor LB-100, and NMDAR inhibitor MK801 were used to investigate the involved in vivo and in vitro pathways. The activation of NMDAR with L-glutamic acid (GLU) or NMDA significantly promoted cellular ferroptosis, iron content, MDA, and the PTGS2 expression, while decreasing GPX4 expression and GSH concentration in human umbilical vein endothelial cells (HUVECs), which was reversed by ferroptosis inhibitors Ferrostatin-1(Fer-1), Liproxstatin-1 (Lip-1), or Deferoxamine (DFO). RNA-seq revealed that ferroptosis and SLC7A11 participate in NMDA or GLU-mediated NMDAR activation. The PP2A-AMPK-HMGB1 pathway was majorly associated with NMDAR activation-induced ferroptosis, validated using the PP2A inhibitor LB-100, AMPK activator AICAR, or HMGB1 siRNA. The role of NMDAR in ferroptosis was validated in HUVECs induced with the ferroptosis activator errasin or RSL3 and counteracted by the NMDAR inhibitor MK-801. The in vivo results showed that NMDA- or GLU-induced ferroptosis and LOS production was reversed by MK-801, LB-100, AICAR, MK-801, and GLY, confirming that the PP2A-AMPK-HMGB1 pathway is involved in NMDAR activation-induced vascular endothelium ferroptosis. In conclusion, the present study demonstrated a novel role of NMDAR in endothelial cell injury by regulating ferroptosis via the PP2A-AMPK-HMGB1 pathway.

Starving the enemy: how plant and microbe compete for sugar on the border.

As the primary energy source for a plant host and microbe to sustain life, sugar is generally exported by Sugars Will Eventually be Exported Transporters (SWEETs) to the host extracellular spaces or the apoplast. There, the host and microbes compete for hexose, sucrose, and other important nutrients. The host and microbial monosaccharide transporters (MSTs) and sucrose transporters (SUTs) play a key role in the "evolutionary arms race". The result of this competition hinges on the proportion of sugar distribution between the host and microbes. In some plants (such as Arabidopsis, corn, and rice) and their interacting pathogens, the key transporters responsible for sugar competition have been identified. However, the regulatory mechanisms of sugar transporters, especially in the microbes require further investigation. Here, the key transporters that are responsible for the sugar competition in the host and pathogen have been identified and the regulatory mechanisms of the sugar transport have been briefly analyzed. These data are of great significance to the increase of the sugar distribution in plants for improvement in the yield.

Corrigendum: Progress in rice sheath blight resistance research.

[This corrects the article DOI: 10.3389/fpls.2023.1141697.].

Exposure to polystyrene nanoparticles at predicted environmental concentrations enhances toxic effects of Acinetobacter johnsonii AC15 infection on Caenorhabditis elegans.

Nanoplastics and microbial pathogens are both widely distributed in the environment; however, their combined toxicity remains largely unclear. Using Caenorhabditis elegans as an animal model, we examined the possible effect of exposure to polystyrene nanoparticle (PS-NP) in Acinetobacter johnsonii AC15 (a bacterial pathogen) infected animals. Exposure to PS-NP at the concentrations of 0.1-10 µg/L significantly enhanced the toxicity of Acinetobacter johnsonii AC15 infection on lifespan and locomotion behaviors. In addition, after exposure to 0.1-10 µg/L PS-NP, the accumulation of Acinetobacter johnsonii AC15 in body of nematodes was also increased. Meanwhile, the innate immune response indicated by the increase of antimicrobial gene expressions in Acinetobacter johnsonii AC15 infected nematodes was suppressed by exposure to 0.1-10 µg/L PS-NP. Moreover, expressions of egl-1, dbl-1, bar-1, daf-16, pmk-1, and elt-2 governing the bacterial infection and immunity in Acinetobacter johnsonii AC15 infected nematodes were further inhibited by exposure to 0.1-10 µg/L PS-NP. Therefore, our data suggested the possible exposure risk of nanoplastic at predicted environmental concentrations in enhancing the toxic effects of bacterial pathogens on environmental organisms.

Transcriptome Analysis Reveals Genes Associated with Flooding Tolerance in Mulberry Plants.

Mulberry (Morus alba), a widely distributed economic plant, can withstand long-term flooding stress. However, the regulatory gene network underlying this tolerance is unknown. In the present study, mulberry plants were subjected to submergence stress. Subsequently, mulberry leaves were collected to perform quantitative reverse-transcription PCR (qRT-PCR) and transcriptome analysis. Genes encoding ascorbate peroxidase and glutathione S-transferase were significantly upregulated after submergence stress, indicating that they could protect the mulberry plant from flood damage by mediating ROS homeostasis. Genes that regulate starch and sucrose metabolism; genes encoding pyruvate kinase, alcohol dehydrogenase, and pyruvate decarboxylase (enzymes involved in glycolysis and ethanol fermentation); and genes encoding malate dehydrogenase and ATPase (enzymes involved in the TCA cycle) were also obviously upregulated. Hence, these genes likely played a key role in mitigating energy shortage during flooding stress. In addition, genes associated with ethylene, cytokinin, abscisic acid, and MAPK signaling; genes involved in phenylpropanoid biosynthesis; and transcription factor genes also showed upregulation under flooding stress in mulberry plants. These results provide further insights into the adaptation mechanisms and genetics of submergence tolerance in mulberry plants and could aid in the molecular breeding of these plants.

Progress in rice sheath blight resistance research.

Rice sheath blight (ShB) disease poses a major threat to rice yield throughout the world. However, the defense mechanisms against ShB in rice remain largely unknown. ShB resistance is a typical quantitative trait controlled by multiple genes. With the rapid development of molecular methods, many quantitative trait loci (QTLs) related to agronomic traits, biotic and abiotic stresses, and yield have been identified by genome-wide association studies. The interactions between plants and pathogens are controlled by various plant hormone signaling pathways, and the pathways synergistically or antagonistically interact with each other, regulating plant growth and development as well as the defense response. This review summarizes the regulatory effects of hormones including auxin, ethylene, salicylic acid, jasmonic acid, brassinosteroids, gibberellin, abscisic acid, strigolactone, and cytokinin on ShB and the crosstalk between the various hormones. Furthermore, the effects of sugar and nitrogen on rice ShB resistance, as well as information on genes related to ShB resistance in rice and their effects on ShB are also discussed. In summary, this review is a comprehensive description of the QTLs, hormones, nutrition, and other defense-related genes related to ShB in rice. The prospects of targeting the resistance mechanism as a strategy for controlling ShB in rice are also discussed.

Seasonal pollution and surface characteristics of microplastics in surface water in the Wanzhou section of the Three Gorges Reservoir, China.

The pollution of freshwater environments with microplastics (MPs) has attracted increasing attention owing to their threats to aquatic ecosystems and human health. Here, we sampled and analyzed MPs from mainstream, tributary, and backwater areas in the Wanzhou section of the Three Gorges Reservoir (TGR) in impoundment and flood periods. Microplastic pollution was the most severe in the backwater areas. The average abundance of MPs reached the highest value in the flood period (5.27±3.47×107 items km-2), which was 3-5 times that in the impoundment period. In the 0.3-5 mm size class, the 1-5 mm fraction was the most abundant, accounting for more than 81% in the flood period and 68% of the total MP particle abundance in the impoundment period in the mainstream and backwater areas. However, 0.3-1 mm MPs contributed more than 50% in the tributaries during the impoundment period. Polystyrene, polypropylene, and polyethylene MPs were detected in foam, fragment, sheet, and line-shaped MP particles. White, opaque, foamed polystyrene MPs contributed 32-81% to total MP particle abundance in the watershed. Microplastic particle surfaces showed signs of damage and oxidation, and ten different elements were found. Oxygen was clustered on the surface of foam and fragment MPs. Microplastic pollution was severe in the Wanzhou watershed. Especially in the backwater areas, oxidized MPs of variable shapes derived mainly from surface runoff in the flood period and sewage discharge in the impoundment period were abundant. The results of this study contribute to understanding seasonal pollution patterns and surface characteristics of MPs in the TGR and similar watersheds.

Evaluation Soybean Cultivars for Reaction to Heterodera glycines Populations HG Types 7 and 1.3.4.7 in Northeast China.

Soybean cyst nematode Heterodera glycines (SCN) is a major threat to global soybean production. Effective management of this disease is dependent on the development of resistant cultivars. Two SCN HG Types, 7 and 1.3.4.7. were previously identified as prevalent H. glycines populations in Northeast China. In order to evaluate soybean cultivars resistant to local SCN populations, 110 domestic commercial soybeans from different regions of Northeast China were assessed in the greenhouse to determine their potential as novel sources of resistance. The results suggested that cultivars responded differently to the two HG types. Of the 110 soybean cultivars evaluated, 24 accessions were classified as resistant or moderately resistant to HG Type 7, and five cultivars were classified as resistant or moderately resistant to HG Type 1.3.4.7. Among the tested cultivars, Kangxian 12 and Qingdou 13 had resistance response to both HG types 7 and 1.3.4.7. Thus, these broad-based SCN cultivars will be the valuable materials in the SCN resistance breeding program.

Health risks of Cryptosporidium and Giardia in the application of surface water and septic tank effluent in Chinese agriculture: Impact on cancer patients identified by quantitative microbial risk assessment.

The protozoa Cryptosporidium and Giardia are major causes of diarrhea and are commonly found on vegetables in China. They pose a health risk, particularly to immunocompromised individuals, including cancer patients. A quantitative microbial risk assessment of Chinese data evaluated the risks of Cryptosporidium and Giardia exposure arising from the application of surface water and septic tank effluent to agricultural land. Exposure via agricultural produce consumption (consumers) and agricultural practices (farmers) was considered for subpopulations of cancer patients and immunocompetent people in urban and rural areas, and risk mitigation scenarios were modelled. The cumulative disease burdens attributable to cryptosporidiosis and giardiasis were, respectively, 9.68×10-6 and 5.57×10-5 disability-adjusted life years per person per year (DALYs pppy) for immunocompetent people, and 3.14×10-5 and 1.51×10-4 DALYs pppy for cancer patients. Cancer patients were approximately three times more likely to have an individual disease burden than immunocompetent people. The disease burden was higher for consumers than farmers, and higher in rural areas than urban areas (all exceeding the maximum recommended by the World Health Organization). The highest burdens were in provinces of high population, such as Henan, Guangdong, and Sichuan, while the burden associated with human and livestock fecal effluent application was higher than with surface water irrigation. Of the three vegetables studied, lettuce posed the greatest risk, followed by bok choy, while cucumber posed the least risk. Risk mitigation scenario analysis showed that pre-treatment of surface water and feces, and appropriate post-harvest handling of vegetables, including disinfection, cooking, and adequate surface heat treatment (75 °C for 60 s), should be considered when attempting to reduce disease burdens. The methodology and findings of this study are useful for evaluating and reducing the burden of Cryptosporidium and Giardia infections associated with agricultural irrigation and fertilization practices, particularly on cancer patients.

Spatially explicit model of the Cryptosporidium and Giardia disease burden from surface and ground waters in urban and rural areas of the Three Gorges Reservoir watershed in Chongqing, China.

Cryptosporidium and Giardia (major causes of diarrhea) are widely distributed in Chinese source waters and threaten human health. A new spatially explicit GloWPa-TGR-Crypt-Giar C1 model is presented to simultaneously estimate mean monthly (oo)cyst concentrations in surface and ground waters in the Three Gorges Reservoir (TGR) watershed. A quantitative risk assessment of protozoal infections considered different source waters, transmission pathways, regions, susceptible subpopulations, and drinking water treatments. Monthly mean Cryptosporidium oocyst and Giardia cyst concentrations ranged between 0.5-19.3 oocysts/10 L and 0.2-5.0 cysts/10 L in surface water, respectively, and 0.007-0.3 oocysts/10 L and 0.002-0. 2 cysts/10 L in groundwater. The cumulative disease burdens attributable to cryptosporidiosis and giardiasis were, respectively, 5.77×10-5 DALYs (disability-adjusted life years/person/year) and 4.63×10-6 DALYs in urban areas, and 6.35×10-4 DALYs and 8.84×10-5 DALYs in rural areas, which were much higher than the reference risk level recommended by the World Health Organization ([Formula: see text] DALYs). The annual burden associated with consuming surface water was calculated to be 3.84×10-4 DALYs for Cryptosporidium and [Formula: see text] DALYs for Giardia, whereas consuming groundwater entailed the lower burdens (1.26×10-5 and 3.50×10-6 DALYs, respectively). Most DALYs were a consequence of consumption of directly supplied surface water. Fifty percent of the health burden was carried by immunodeficiency with HIV. Children (0-4 years) were more likely to have an individual disease burden than adults (15-64 years). Males were more susceptible than females. Improving sanitation through adequate ozone and microfiltration treatment should be considered when attempting to reduce disease burden. Sensitivity analysis highlighted the importance of reducing (oo)cyst loads to protect the watershed. The methodology and results described will help in evaluating and reducing the burden of protozoal infection associated with surface and ground waters in the TGR and similar watersheds.

Exposure to Nanoplastic Particles Enhances Acinetobacter Survival, Biofilm Formation, and Serum Resistance.

The interaction between nanoplastics and bacteria remains still largely unclear. In this study, we determined the effect of nanopolystyrene particle (NP) on a bacterial pathogen of Acinetobacter johnsonii AC15. Scanning electron microscopy (SEM) analysis indicated the aggregation of NPs from 10 µg/L to 100 µg/L on surface of A. johnsonii AC15, suggesting that A. johnsonii AC15 acted as the vector for NPs. Exposure to 100−1000 µg/L NPs increased the growth and colony-forming unit (CFU) of A. johnsonii AC15. In addition, exposure to 100−1000 µg/L NPs enhanced the amount of formed biofilm of A. johnsonii AC15. Alterations in expressions of 3 survival-related (zigA, basD, and zur), 5 biofilm formation-related (ompA, bap, adeG, csuC, and csuD), and 3 serum resistance-related virulence genes (lpxC, lpxL, and pbpG) were observed after exposure to 1000 µg/L NPs. Moreover, both CFU and survival rate of A. johnsonii AC15 in normal human serum (NHS) were significantly increased by 1−1000 µg/L NPs, suggesting the enhancement in serum resistance of Acinetobacter pathogen by NPs. In the NHS, expressions of 3 survival-related (zigA, basD, and zur), 9 biofilm formation-related (ompA, bap, adeF, adeG, csuA/B, csuC, csuD, csuE, and hlyD), and 3 serum resistance-related virulence genes (lpxC, lpxL, and pbpG) were affected by 1000 µg/L NPs. Expressions of 1 survival-related (zigA), 5 biofilm formation-related (bap, adeG, csuC, csuD, and csuE), and 3 serum resistance-related virulence genes (lpxC, lpxL, and pbpG) were also altered by 10 µg/L NPs after the addition of NHS. Therefore, exposure to NPs in the range of µg/L has the potential to enhance bacterial virulence by increasing their growth, biofilm formation, and serum resistance.

Quantitative Risk Assessment of Vibrio parahaemolyticus in Shellfish from Retail to Consumption in Coastal Cities of Eastern China.

ABSTRACT: Vibrio parahaemolyticus is the main foodborne pathogen worldwide that causes acute gastroenteritis. A quantitative microbiological risk assessment (QMRA) was conducted to evaluate the health risk associated with V. parahaemolyticus in shellfish in the coastal cities in the eastern part of the People's Republic of China. The QMRA framework was established from shellfish at retail to cooking at home to consumption. The prevalence and level of V. parahaemolyticus in shellfish, cooking methods, storage temperature, time after purchase, shellfish consumption frequency, and consumption amount were analyzed in the exposure assessment. The results of the exposure assessment were introduced into the beta-Poisson dose-response model, and Monte Carlo analysis was used to calculate the risk of gastroenteritis from shellfish consumption. The probability of illness caused by V. parahaemolyticus from shellfish consumption per person per year (Pill,yr) was 3.49E-05. Seasonal differences were noted in the Pill/meal; the maximum was 4.81E-06 in summer and the minimum was 2.27E-07 in winter. The sensitivity analysis revealed that the level of V. parahaemolyticus in shellfish and the amount of shellfish consumed per meal were main factors contributing to illness. This QMRA provided valuable information such as the probability of illness associated with the consumption of shellfish and reference points for prevention strategies and control standards of V. parahaemolyticus in shellfish.

Concanavalin A promotes angiogenesis and proliferation in endothelial cells through the Akt/ERK/Cyclin D1 axis.

CONTEXT: Concanavalin A (Con A) exhibited multiple roles in cancer cells. However, the role of Con A in endothelial cells was not reported. OBJECTIVE: Our present study investigated the potential angiogenic role of Con A in endothelial cells and ischaemic hind-limb mice. MATERIALS AND METHODS: Human umbilical vein endothelial cells and Ea.hy926 cells were employed to determine the effect of Con A (0.3, 1, and 3 µg/mL) or vehicle on angiogenesis and cell proliferation with tube formation, ELISA, flow cytometry, EdU, and western blot. Hind-limb ischaemic mice were conducted to determine the pro-angiogenic effect of Con A (10 mg/kg) for 7 days. RESULTS: Con A promoted tube formation to about three-fold higher than the control group and increased the secretion of VEGFa, PDGFaa, and bFGF in the medium. The cell viability was promoted to 1.3-fold by Con A 3 µg/mL, and cell cycle progression of G0G1 phase was decreased from 77% in the vehicle group to 70% in Con A 3 µg/mL, G2M was promoted from 15 to 19%, and S-phase was from 7 to 10%. Con A significantly stimulated phosphorylation of Akt and ERK1/2 and expression of cyclin D1 and decreased the expression of p27. These effects of Con A were antagonised by the PI3K inhibitor LY294002 (10 µM) and MEK pathway antagonist PD98059 (10 µM). Moreover, Con A (10 mg/kg) exhibited a repair effect in ischaemic hind-limb mice. DISCUSSION AND CONCLUSIONS: This study will provide a new option for treating ischaemic disease by local injection with Con A.

Biosafety assessment of Acinetobacter strains isolated from the Three Gorges Reservoir region in nematode Caenorhabditis elegans.

Acinetobacter has been frequently detected in backwater areas of the Three Gorges Reservoir (TGR) region. We here employed Caenorhabditis elegans to perform biosafety assessment of Acinetobacter strains isolated from backwater area in the TGR region. Among 21 isolates and 5 reference strains of Acinetobacter, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii ATCC 19606T, A. junii NH88-14, and A. lwoffii DSM 2403T resulted in significant decrease in locomotion behavior and reduction in lifespan of Caenorhabditis elegans. In nematodes, exposure to Acinetobacter strains of AC1, AC15, AC18, AC21, A. baumannii, A. junii and A. lwoffii also resulted in significant reactive oxygen species (ROS) production. Moreover, exposure to Acinetobacter isolates of AC1, AC15, AC18, and AC21 led to significant increase in expressions of both SOD-3::GFP and some antimicrobial genes (lys-1, spp-12, lys-7, dod-6, spp-1, dod-22, lys-8, and/or F55G11.4) in nematodes. The Acinetobacter isolates of AC1, AC15, AC18, and AC21 had different morphological, biochemical, phylogenetical, and virulence gene properties. Our results suggested that exposure risk of some Acinetobacter strains isolated from the TGR region exists for environmental organisms and human health. In addition, C. elegans is useful to assess biosafety of Acinetobacter isolates from the environment.

OsLEA1a overexpression enhances tolerance to diverse abiotic stresses by inhibiting cell membrane damage and enhancing ROS scavenging capacity in transgenic rice.

Late embryogenesis abundant (LEA) proteins are involved in diverse abiotic stresses tolerance in many different organisms. Our previous studies have shown that the heterologous expression of OsLEA1a interfered with the resistance of Escherichia coli to abiotic stresses. However, in the present study, based on growth status and physiological indices of rice plant, the overexpression of OsLEA1a in rice conferred increased resistance to abiotic stresses compared with the wild-type (WT) plants. Before applying abiotic stresses, there were no significant differences in physiological indices of rice seedlings. After NaCl, sorbitol, CuSO4 and H2O2 stresses, the transgenic lines had lower relative electrical conductivity, malondialdehyde and lipid peroxidation, greater the contents of proline, soluble sugar and glutathione, and higher the activities of superoxide dismutase, catalase and peroxidase than the WT plants. The results indicate that the OsLEA1a gene is involved in the protective response of plants to various abiotic stresses by inhibiting cell membrane damage and enhancing reactive oxygen species scavenging capacity. It was speculated that post-translational modification causes OsLEA1a functional differences in E. coli and rice. The present study shows that OsLEA1a could be a useful candidate gene for engineering abiotic stress tolerance in cultivated plants.