Development of a CRISPR/Cpf1 system for multiplex gene editing in Aspergillus oryzae.

CRISPR/Cas technology is a powerful tool for genome engineering in Aspergillus oryzae as an industrially important filamentous fungus. Previous study has reported the application of the CRISPR/Cpf1 system based on the Cpf1 (LbCpf1) from Lachnospiraceae bacterium in A. oryzae. However, multiplex gene editing have not been investigated using this system. Here, we presented a new CRISPR/Cpf1 multiplex gene editing system in A. oryzae, which contains the Cpf1 nuclease (FnCpf1) from Francisella tularensis subsp. novicida U112 and CRISPR-RNA expression cassette. The crRNA cassette consisted of direct repeats and guide sequences driven by the A. oryzae U6 promoter and U6 terminator. Using the constructed FnCpf1 gene editing system, the wA and pyrG genes were mutated successfully. Furthermore, simultaneous editing of wA and pyrG genes in A. oryzae was performed using two guide sequences targeting these gene loci in a single crRNA array. This promising CRISPR/Cpf1 genome-editing system provides a powerful tool for genetically engineering A. oryzae.

Co-digestion of chicken manure and sewage sludge in black soldier fly larvae bioconversion system: bacterial biodiversity and nutrients quality of residues for biofertilizer application.

Black soldier fly larvae (BSFL) bioconversion system is emerging as an effective approach for organic waste pollution treatment. Co-digestion of different organic matters with BSFL can be an effective way to realize the innovative biowaste circular economy. In this study, organic waste mixture of chicken manure and sewage sludge was chosen as substrate for BSFL growth. The bacterial biodiversity and nutrients quality of BSFL residue were evaluated through gene sequencing and other characterizations to confirm their application potential as biofertilizers. The dominant bacteria in BSFL residue were Firmicutes (75.39%) at phylum level, Bacilli (71.61%) at class level and Pseudogracilibacillus (11.08%) at genus level. Antibiotic resistance genes (ARGs) were used to assess the harmlessness of BSFL residue. After BSFL treatment, 36.2% decrease in ARGs was observed. Taking nutrients quality into consideration, dissolved organic carbon, dissolved nitrogen, available phosphorous, and available potassium significantly increased in the co-digestion system. These results demonstrated that co-digestion of chicken manure and excess sludge in BSFL bioconversion system could improve the nutrients quality of residues. However, removal of ARGs in the bioconversion process should be further explored to eliminate environmental concerns associated with application of BSFL residue as biofertilizers.

Characteristics and potential human health risks of microplastics identified in typical clams from South Yellow Sea Mudflat.

The ingestion of clams (Meretrix) with microplastics (MP) contamination could pose potential risk to human health. The characteristics and potential risks of MP identified in wild-clam and farm-clam from South Yellow Sea Mudflat were studied comprehensively in this paper. The results indicated that MP were identified in both wild-clam (3.4-21.3 items/individual, 2.11-10.65 items/g) and farm-clam (1.3-20.8 items/individual, 0.62-8.67 items/g) among 21 sampling sites along South Yellow Sea Mudflat. The MP abundance of clams from marine estuarine or coast ports were significantly higher than those from purely marine coast mudflat, implying that environmental habitats played an important role on MP characteristics. MP abundance were significantly and positively related to shell length, shell height, shell width and soft tissue wet weight by Pearson test, suggesting the bigger the shell, there existed more MP abundance. Among MP in wild-clams and farm-clams, fragment, fiber were most abundant MP shapes, most MP's sizes were lower than 0.25 mm, the predominant colors were black, red, blue and transparent, chlorinated polyethylene (CPE) was the major polymer. Additionally, estimated dietary intake (EDI) of MP for adults via consumption of wild-clam and farm-clam were 1123.33 ± 399.97 and 795.07 ± 326.72 items/kg/year, respectively, suggesting EDI values of wild-clams were higher than those of farm-clams, and MP intake via wild-clam consumption were more than that via farm-clam consumption. The polymer risk indexes (PRI) of MP in total tissue and digestive system for wild-clam were 1297.8 ± 92.15 (hazard level: IV ~ V), 1038 ± 69.55 (IV ~ V), respectively, while PRI of MP in total tissue and digestive system for farm-clam were 979.92 ± 75.45 (III ~ IV), 735 ± 47.78 (III ~ IV), respectively, implying that PRI and hazard level of MP from wild-clam were higher than those from farm-clam, and the potential risks would decrease greatly when digestive systems of clams are removed during ingestion.

Functional role of a novel zinc finger protein, AoZFA, in growth and kojic acid synthesis in Aspergillus oryzae.

Kojic acid (KA) is a valuable secondary metabolite that is regulated by zinc finger proteins in Aspergillus oryzae. However, only two such proteins have been characterized to function in kojic acid production of A. oryzae to date. In this study, we identified a novel zinc finger protein, AoZFA, required for kojic acid biosynthesis in A. oryzae. Our results showed that disruption of AozfA led to increased expression of kojA and kojR involved in kojic acid synthesis, resulting in enhanced kojic acid production, while overexpression of AozfA had the opposite effect. Furthermore, deletion of kojR in the AozfA disruption strain abolished kojic acid production, whereas overexpression of kojR enhanced it, indicating that AoZFA regulates kojic acid production by affecting kojR. Transcriptional activation assay revealed that AoZFA is a transcriptional activator. Interestingly, when kojR was overexpressed in the AozfA overexpression strain, the production of kojic acid failed to be rescued, suggesting that AozfA plays a distinct role from kojR in kojic acid biosynthesis. Moreover, we found that AozfA was highly induced by zinc during early growth stages, and its overexpression inhibited the growth promoted by zinc, whereas its deletion had no effect, suggesting that AoZFA is non-essential but has a role in the response of A. oryzae to zinc. Overall, these findings provide new insights into the roles of zinc finger proteins in the growth and kojic acid production of A. oryzae.IMPORTANCEKojic acid (KA) is an economically valuable secondary metabolite produced by Aspergillus oryzae due to its vast biological activities. Genetic modification of A. oryzae has emerged as an efficient strategy for enhancing kojic acid production, which is dependent on the mining of genes involved in kojic acid synthesis. In this study, we have characterized a novel zinc-finger protein, AoZFA, as a negative regulator of kojic acid production by affecting kojR. AozfA is an excellent target for improving kojic acid production without any effects on the growth of A. oryzae. Furthermore, the simultaneous modification of AozfA and kojR exerts a more significant promotional effect on kojic acid production than the modification of single genes. This study provides new insights for the regulatory mechanism of zinc finger proteins in the growth and kojic acid production of A. oryzae.

Indoor environment in relation to recurrent childhood asthma in Yancheng, China: a hospital-based case-control study.

This investigation explored the association between indoor environmental factors and childhood asthma in Yancheng, China. Asthma case (201 children with recurrent asthma) and control cohorts (242 healthy subjects) were recruited from a Traditional Chinese Medical (TCM) Hospital in Yancheng city, based on the results of an ISAAC questionnaire. Questionnaires regarding environmental risk factors were completed by the child's primary caregivers. To compare data on environmental VOCs and formaldehyde contents between asthma and control cohorts, we passively conducted a 10-day indoor and outdoor sampling. Breastfeeding was a major protective indoor environmental factor for recurrent asthma (adjusted odds ratio [aOR]: 0.368, 95% confidence interval [CI]: 0.216-0.627). Our analysis revealed that childhood recurrent asthma was intricately linked to a family history of asthma. Recurrent asthma was also associated with passive smoking [aOR2.115 (95%-CI 1.275-3.508)]. Analogous correlations were observed between household renovation or new furniture introduction and recurrent asthma [aOR3.129(95%-CI1.542-6.347)]. Benzene and formaldehyde were present in all examined homes. Enhanced benzene and formaldehyde concentrations were strongly evident among asthma versus control cohorts, and they were strongly correlated with augmented recurrent asthma risk. Home environment heavily regulates incidences of childhood recurrent asthma. Hence, actions against the indoor environmental risk factors described in this study may assist in the prevention of recurrent asthma among children.

Pathogenesis diagnosis of a pediatric patient suffering from multi-organ abscesses.

A pediatric patient suffered from renal and hepatic abscesses (through hepatorenal space) and recovered by antibiotics and other therapies. By clinical analysis, the multi-organ abscesses might be caused by bloodstream-disseminated infection. In order to identify the pathogen, we collected kidney biopsy tissue, swabs, and plasma samples, and used metagenomics next-generation sequencing (mNGS) and some traditional methods. The results revealed that polymicrobial especially anaerobic bacteremia (Bacteroides fragilis, et al.) contributed to the abscess formation. What is more, systematic human adenovirus C (HAdV-C) infection was shown, and the virus was isolated. The titer of HAdV-2 neutralizing antibodies was 1/4 in the plasma after symptoms onset. Although the exact mechanism of HAdV-2 infection in multiple abscess formation has not been clarified, the case of multi-organ abscesses in the context of polymicrobial especially anaerobic bacteremia and HAdV infection in healthy children is infrequent.

Pediatric deep venous thrombosis associated with Staphylococcus aureus osteomyelitis.

INTRODUCTION: Our objective was to evaluate clinical features of children with deep venous thrombosis (DVT) and acute hematogenous osteomyelitis (AHO) caused by Staphylococcus aureus. METHODOLOGY: We analyzed 4 years of medical records of patients with AHO and DVT caused by Staphylococcus aureus (S. aureus) and compared clinical and biochemical characteristics of AHO with and without DVT, as well as patients whose DVT dissolved in ≥ 3 weeks. RESULTS: DVT was found in 19/87 AHO individuals (22%). The median age was 9 years (range: 0.5-15 years). 74% (14/19) patients were boys. Methicillin-susceptible Staphylococcus aureus (MSSA) was present in 58% (11/19) cases. The femoral vein and common femoral vein were the two most damaged veins (9 cases each). Anticoagulation therapy with low molecular weight heparin was given to 18 (95%) patients. Within 3 weeks of anticoagulation, 7/13 (54%) with available data had completely resolved DVT. There was no rehospitalization due to bleeding or recurrent DVT. Patients with DVT were found to be older and had increased levels of C-reactive protein, procalcitonin, D-dimer, positive blood culture, incidence of intensive care unit admission, multifocal rate, and length of hospital stay. We did not find clinical difference between patients whose DVT dissolved within 3 weeks and those with > 3 weeks. CONCLUSIONS: Over 20% of patients with S. aureus AHO developed DVT. MSSA accounted for more than half of the cases. DVT was completely resolved in more than half of the cases after 3 weeks of anticoagulant medication, with no sequelae.

The C2H2-type zinc-finger regulator AoKap5 is required for the growth and kojic acid synthesis in Aspergillus oryzae.

Aspergillus oryzae is an important filamentous fungus widely used for the industrial production of fermented foods and secondary metabolites. The clarifying of the mechanism of the growth and secondary metabolites in A. oryzae is important for its industrial production and utilization. Here, the C2H2-type zinc-finger protein AoKap5 was characterized to be involved in the growth and kojic acid production in A. oryzae. The Aokap5-disrupted mutants were constructed by the CRISPR/Cas9 system, which displayed increased colony growth but decreased conidial formation. Deletion of Aokap5 enhanced the tolerance to cell-wall and oxidative but not osmotic stress. The transcriptional activation assay revealed that AoKap5 itself didn't have transcriptional activation activity. Disruption of Aokap5 resulted in the reduced production of kojic acid, coupled with the reduced expression of the kojic acid synthesis genes kojA and kojT. Meanwhile, overexpression of kojT could rescue the decreased production of kojic acid in Aokap5-deletion strain, indicating that Aokap5 serves upstream of kojT. Furthermore, the yeast one-hybrid assay demonstrated that AoKap5 could directly bind to the kojT promoter. These findings suggest that AoKap5 regulates kojic acid production through binding to the kojT promoter. This study provides an insight into the role of zinc finger protein in the growth and kojic acid biosynthesis of A. oryzae.

Identification and evaluation of fishy odorants produced by four algae separated from drinking water source during low temperature period: Insight into odor characteristics and odor contribution of fishy odor-producing algae.

Disgusting fishy odor could break out inside oligotrophic drinking waterbody in winter with low temperature. Nevertheless, fishy odor-producing algae and corresponding fishy odorants were not very clear, odor contribution of fishy odorant and odor-producing algae to overall odor profile were also not well understood. In this study, the fishy odorants, produced by four algae separated from Yanlong Lake, were identified simultaneously. Odor contribution of identified odorant, separated algae to overall fishy odor profile were both evaluated. The results indicated Yanlong Lake was mainly associated with fishy odor (flavor profile analysis (FPA) intensity: 6), eight, five, five and six fishy odorants were identified and determined in Cryptomonas ovate, Dinobryon sp., Synura uvella, Ochromonas sp., respectively, which were separated and cultured from water source. Totally sixteen odorants with concentration range of 90-880 ng/L, including hexanal, heptanal, 2,4-heptadienal, 1-octen-3-one, 1-octen-3-ol, octanal, 2-octenal, 2,4-octadienal, nonanal, 2-nonenal, 2,6-nonadienal, decanal, 2-decenal, 2,4-decadienal, undecanal, 2-tetradecanone, were verified in separated algae and associated with fishy odor. Although more odorants' odor activity value (OAV) were lower than one, approximately 89%, 91%, 87%, 90% of fishy odor intensities could be explained by reconstituting identified odorants for Cryptomonas ovate, Dinobryon sp., Synura uvella, Ochromonas sp., respectively, suggesting synergistic effect could exist among identified odorants. By calculating and evaluating total odorant production, total odorant OAV and cell odorant yield of separated algae, odor contribution rank to overall fishy odor should be Cryptomonas ovate (28.19%), Dinobryon sp. (27.05%), Synura uvella (24.27%), Ochromonas sp. (20.49%). This is the first study for identifying fishy odorants from four actually separated odor-producing algae simultaneously, this is also for the first time evaluating and explaining odor contribution of identified odorant, separated algae to overall odor profile comprehensively, this study will supply more understanding for controlling and managing fishy odor in drinking water treatment plant.

A critical review on graphene oxide membrane for industrial wastewater treatment.

An important way to promote the environmental industry's goal of carbon reduction is to promote the recycling of resources. Membrane separation technology has unique advantages in resource recovery and advanced treatment of industrial wastewater. However, the great promise of traditional organic membrane is hampered by challenges associated with organic solvent tolerance, lack of oxidation resistance, and serious membrane fouling control. Moreover, the high concentrations of organic matter and inorganic salts in the membrane filtration concentrate also hinder the wider application of the membrane separation technology. The emerging cost-effective graphene oxide (GO)-based membrane with excellent resistance to organic solvents and oxidants, more hydrophilicity, lower membrane fouling, better separation performance has been expected to contribute more in industrial wastewater treatment. Herein, we provide comprehensive insights into the preparation and characteristic of GO membranes, as well as current research status and problems related to its future application in industrial wastewater treatment. Finally, concluding remarks and future perspectives have been deduced and recommended for the GO membrane separation technology application for industrial wastewater treatment, which leads to realizing sustainable wastewater recycling and a nearly "zero discharge" water treatment process.

Efficient biodegradation characteristics and detoxification pathway of organophosphorus insecticide profenofos via Cupriavidus nantongensis X1T and enzyme OpdB.

Profenofos residues in the environment pose a high risk to mammals and non-target organisms. In this study, the biodegradation and detoxification of profenofos in an efficient degrading strain, Cupriavidus nantongensis X1T, was investigated. Strain X1T could degrade 88.82 % of 20 mg/L profenofos in 48 h. The optimum temperature and inoculation amount of strain X1T for the degradation of profenofos were 30-37 °C and 20 % (V/V), respectively. Metabolic pathway analysis showed that strain X1T could degrade both profenofos and its main metabolite 4-bromo-2-chlorophenol. Metabolite toxicity analysis results showed that dehalogenation was the main detoxification step in profenofos biodegradation. The key gene and enzyme for profenofos degradation in strain X1T were also explored. RT-qPCR shows that organophosphorus hydrolase (OpdB) was the key enzyme to control the hydrolysis process in strain X1T. The purified enzyme OpdB in vitro had the same degradation characteristics as strain X1T. Divalent metal cations could significantly enhance the hydrolysis activity of strain X1T and enzyme OpdB. Meanwhile, strain X1T could degrade 60.89 % of 20 mg/L profenofos in actual field soil within 72 h. This study provides an efficient biological resource for the remediation of profenofos residual pollution in the environment.

Effects of different cathodic potentials on performance, microbial community structure and function for bioelectrochemical-stimulated dechlorination of 2,4,6-trichlorophenol in sediments.

Bioelectrochemical systems with biocathodes constitute a promising means to enhance the biological dechlorination of 2,4,6-trichlorophenol (2,4,6-TCP) in constructed wetland (CW) sediments. However, the effect of different cathodic potentials on the structure and function of 2,4,6-TCP-reducing biocathode communities in CW sediments is largely unknown. Here, we evaluated the performance and microbial community structure of 2,4,6-TCP-reducing biocathode systems at different cathodic potentials (- 0.5, - 0.7, - 0.9, and - 1.1 V vs. saturated calomel electrode). The dechlorination efficiency of 2,4,6-TCP with the biocathode relatively increased by 16.02%-33.17% compared to that in the open circuit. The highest 2,4,6-TCP dechlorination efficiency (92.34 ± 0.86%) was observed at - 0.7 V in sediment, which may be due to the highest abundance of functional genera (e.g., Pseudomonas, Spirochaeta) at - 0.7 V. Metagenomic analysis provided new insights into the metabolic potential of microorganisms in CW sediments and suggested possible 2,4,6-TCP conversion pathways in sediments. 2,4,6-TCP was gradually dechlorinated to form 4-chlorophenol, followed by a ring-opening step via the activities of chlorophenol reductive dehalogenase and oxygenase (e.g., cprA, tfdB). Interestingly, micro-electrical stimulation enhanced the expression of chlorophenol reductive dehalogenase (cprA). Therefore, our findings at the molecular and gene expression levels provide insights into the effects of different cathodic potentials on the performance and community structure of 2,4,6-TCP-reducing biocathode systems in CW sediments.

Effect of different carbon sources on sulfate reduction and microbial community structure in bioelectrochemical systems.

Microbial electrolysis cells (MECs) have rapidly developed into a promising technology to treat sulfate-rich wastewater that lacks electron donors. Hence, a better understanding of the effect on the microbial community structure caused by different sources in bioelectrochemical systems is required. This study sought to investigate the effect of different carbon sources (NaHCO3, ethanol, and acetate were employed as sole carbon source respectively) on the performance of sulfate-reducing biocathodes. The sulfate reduction efficiency enhanced by the bioelectrochemical systems was 8.09 - 11.57% higher than that of open-circuit reference experiments. Furthermore, the optimum carbon source was ethanol with a maximum sulfate reduction rate of 170 mg L-1 d-1 in the bioelectrochemical systems. The different carbon sources induced significant differences in sulfate reduction efficiency as demonstrated by the application of a micro-electrical field. Microbial community structure and network analysis revealed that all three kinds of carbon source systems enriched large proportions of sulfate-reducing bacteria and electroactive bacteria but were significantly distinct in composition. The dominant sulfate-reducing bacteria that use NaHCO3 and acetate as carbon sources were Desulfobacter and Desulfobulbus, whereas those that use ethanol as carbon source were Desulfomicrobium and Desulfovibrio. Our results suggest that ethanol is a more suitable carbon source for sulfate reduction in bioelectrochemical systems.

Clinical characteristics of central nervous system candidiasis due to Candida albicans in children: a single-center experience.

BACKGROUND: Central nervous system candidiasis due to Candida albicans (CNSC) in children is easily misdiagnosed and is associated with poor outcomes and a high mortality rate. There is no big data research or systematic review of CNSC. METHODS: Patients diagnosed as CNSC with positive culture results of Candida albicans in Beijing Children's Hospital affiliated to Capital Medical University from March 2010 to March 2019 were included. Patients receiving immunosuppressive therapy or transplantation, or with malignant tumours were excluded. We analysed the clinical characteristics, follow-up results, drug susceptibility tests and whole-exome sequencing (WES) results. RESULTS: Thirty-three definitive patients were enrolled, including 22 males and 11 females. Twenty-five patients suffered from CNSC when they were less than 1 year old, and a total of 29 patients had high-risk factors. The main clinical manifestations were fever, convulsions, and positive neurological signs. Twenty-two patients had CNS infections alone, and 11 patients had CNS infections combined with invasive infections involving multiple sites. Twenty-seven cases had a positive CSF and/or blood culture at our hospital. All strains were susceptible to fluconazole, and 2 strains had intermediate susceptibility to voriconazole. As for amphotericin B, all the strains were wild type (WT). WES of 16 patients revealed 2 cases with CARD9 mutations, who suffered from recurrent onychomycosis or thrush before. CONCLUSION: CNSC mostly existed in children younger than 1 year old, who all had underlying risk factors. CNSC patients with onset at an older age or with recurrent superficial fungal infections might have primary immunodeficiency.

MRI-guided microwave ablation and albumin-bound paclitaxel for lung tumors: Initial experience.

Magnetic resonance-guided microwave ablation (MRI-guided MWA) is a new, minimally invasive ablation method for cancer. This study sought to analyze the clinical value of MRI-guided MWA in non-small cell lung cancer (NSCLC). We compared the precision, efficiency, and clinical efficacy of treatment in patients who underwent MRI-guided MWA or computed tomography (CT)-guided microwave ablation (CT-guided MWA). Propensity score matching was used on the prospective cohort (MRI-MWA group, n = 45) and the retrospective observational cohort (CT-MWA group, n = 305). To evaluate the advantages and efficacy of MRI-guided MWA, data including the accuracy of needle placement, scan duration, ablation time, total operation time, length of hospital stay, progression-free survival (PFS), and overall survival (OS) were collected and compared between the two groups. The mean number of machine scans required to adjust the needle position was 7.62 ± 1.69 (range 4-12) for the MRI-MWA group and 9.64 ± 2.14 (range 5-16) for the CT-MWA group (p < 0.001). The mean time for antenna placement was comparable between the MRI and CT groups (54.41 ± 12.32 min and 53.03 ± 11.29 min, p = 0.607). The microwave ablation time of the two groups was significantly different (7.62 ± 2.65 min and 9.41 ± 2.86 min, p = 0.017), while the overall procedure time was comparable (91.28 ± 16.69 min vs. 93.41 ± 16.03 min, p = 0.568). The overall complication rate in the MRI-MWA group was significantly lower than in the CT-MWA group (12% vs. 51%, p = 0.185). The median time to progression was longer in the MRI-MWA group than in the CT-MWA group (11 months [95% CI 10.24-11.75] vs. 9 months [95% CI 8.00-9.99], p = 0.0003; hazard ratio 0.3690 [95% CI 0.2159-0.6306]). OS was comparable in both groups (MRI group 26.0 months [95% CI 25.022-26.978] vs. CT group 23.0 months [95% CI 18.646-27.354], p = 0.18). This study provides hitherto-undocumented evidence of the clinical effects of MRI-guided MWA on patients with NSCLC and determines the relative safety and efficiency of MRI- and CT-guided MWA.

Predictive values of the SOFA score and procalcitonin for septic shock after percutaneous nephrolithotomy.

To investigate the value of combination of the Sequential Organ Failure Assessment (SOFA) score and procalcitonin (PCT) for prediction of septic shock after percutaneous nephrolithotomy (PCNL). A total of 1328 patients receiving PCNL for renal calculi were allocated into control group (without septic shock) and septic shock group, and related data were retrospectively collected. Univariate analysis was firstly performed, and the variables with two sided P < 0.10 were then included in logistic regression analysis to determine independent risk factors. Receiver operating characteristic (ROC) curve was utilized to evaluate the predictive values. Area under curve (AUC) was compared using Z test. Postoperative septic shock was developed in 61 patients (4.6%) and not developed in 1267 patients (95.3%). Multivariate analysis demonstrated that SOFA score (OR: 1.316, 95% CI 1.125-1.922), PCT (OR: 1.205, 95% CI 1.071-1.696) and operative time (OR: 1.108, 95% CI 1.032-1.441) were independent risk factors for septic shock with adjustment for sex, history of urolithiasis surgery, positive history of urine culture and history of PCNL. The ROC curves demonstrated that the AUCs of SOFA score and PCT for predicting septic shock after PCNL were 0.896 (95% CI 0.866-0.927) and 0.792 (95% CI 0.744-0.839), respectively. The AUC of their combination was 0.971 (95% CI 0.949-0.990), which was higher than those of individual predictions (vs 0.896, Z = 4.086, P < 0.001; vs 0.792, Z = 6.983, P < 0.001). Both the SOFA score and PCT could be applied in predicting septic shock after PCNL, and their combination could further elevate the diagnostic ability.

Simultaneous recovery of bio-sulfur and bio-methane from sulfate-rich wastewater by a bioelectrocatalysis coupled two-phase anaerobic reactor.

The microbial electrolysis cell coupled the two-phase anaerobic digestion (MEC-TPAD) was developed for simultaneous recovery of bio-sulfur and bio-methane from sulfate-rich wastewater. In acidogenic phase, the produced sulfides were efficiently converted into bio-sulfur via anodic bio-oxidation, with a maximum recovery of 59 ± 5.5 %. The anode coupled acidogenesis produced more volatile fatty acids which were benefit for the subsequent methanogenesis. The cathode in methanogenic phase created a suitable pH condition and enhanced the methanogenesis. Correspondingly, the maximum bio-methane yield in MEC-TPAD was 2 times higher than that in TPAD. Microbial communities revealed that major functional consortia capable of sulfides oxidation (e.g. Alcaligenes) in anode biofilm, hydrogenotrophic methanogenesis (e.g. Methanobacterium) in cathode biofilm, and acetotrophic methanogenesis (e.g. Methanosaeta) in methanogenic sludge were enriched. Economic benefit could totally cover the cost of input electric energy. This work opens an appealing avenue for recovering nutrient and energy from wastewater.

Recent advances in carrageenan-based films for food packaging applications.

In order to solve the increasingly serious environmental problems caused by plastic-based packaging, carrageenan-based films are drawing much attentions in food packaging applications, due to low cost, biodegradability, compatibility, and film-forming property. The purpose of this article is to present a comprehensive review of recent developments in carrageenan-based films, including fabrication strategies, physical and chemical properties and novel food packaging applications. Carrageenan can be extracted from red algae mainly by hydrolysis, ultrasonic-assisted and microwave-assisted extraction, and the combination of multiple extraction methods will be future trends in carrageenan extraction methods. Carrageenan can form homogeneous film-forming solutions and fabricate films mainly by direct coating, solvent casting and electrospinning, and mechanism of film formation was discussed in detail. Due to the inherent limitations of the pure carrageenan film, physical and chemical properties of carrageenan films were enhanced by incorporation with other compounds. Therefore, carrageenan-based films can be widely used for extending the shelf life of food and monitoring the food freshness by inhibiting microbial growth, reducing moisture loss and the respiration, etc. This article will provide useful guidelines for further research on carrageenan-based films.

High-throughput sequencing unravels the cell heterogeneity of cerebrospinal fluid in the bacterial meningitis of children.

Bacterial meningitis (BM) is a common life-threatening infection in children that occurs in the central nervous system (CNS). The cytologic examination of cerebrospinal fluid (CSF) is a key parameter in the diagnosis of BM, but the heterogeneity of cells in the CSF has not been elucidated, which limits the current understanding of BM neuroinflammation. In this study, CSF samples were collected from a number of BM patients who were in different stages of disease progression. Single-cell RNA-sequencing (scRNA-seq), with additional bulk transcriptome sequencing, was conducted to decipher the characteristics of CSF cells in BM progression. A total of 18 immune cell clusters in CSF were identified, including two neutrophils, two monocytes, one macrophage, four myeloid dendritic cells, five T cells, one natural killer cell, one B cell, one plasmacytoid dendritic cell, and one plasma cell subtype. Their population profiles and dynamics in the initial onset, remission, and recovery stages during BM progression were also characterized, which showed decreased proportions of myeloid cells and increased proportions of lymphoid cells with disease progression. One novel neutrophil subtype, FFAR2+TNFAIP6+ neutrophils, and one novel monocyte subtype, THBS1+IL1B+ monocytes, were discovered, and their quantity changes positively correlated with the intensity of the inflammatory response in the CSF during BM. In addition, the CSF of BM patients with unsatisfactory therapeutic responses presented with different cell heterogeneity compared to the CSF of BM patients with satisfactory therapeutic responses, and their CSF featured altered intercellular communications and increased proportions of type II myeloid dendritic cells and plasmacytoid dendritic cells. Moreover, the bulk transcriptome profiles of autologous CSF cells and peripheral blood leukocytes of BM patients showed that the immune cells in these two physiological compartments exhibited distinct immune responses under different onset conditions. In particular, the CSF cells showed a high expression of macrophage characteristic genes and a low expression of platelet characteristic genes compared with peripheral blood leukocytes. Our study conducted an in-depth exploration of the characteristics of CSF cells in BM progression, which provided novel insights into immune cell engagement in acute CNS infection.