ROK family regulator NagC promotes prodigiosin biosynthesis independent of N-acetylglucosamine in Serratia sp. ATCC 39006.

Serratia sp. ATCC 39006 is an important model strain for the study of prodigiosin production, whose prodigiosin biosynthesis genes (pigA-O) are arranged in an operon. Several transcription factors have been shown to control the transcription of the pig operon. However, since the regulation of prodigiosin biosynthesis is complex, the regulatory mechanism for this process has not been well established. In most γ-proteobacteria, the ROK family regulator NagC acts as a global transcription factor in response to N-acetylglucosamine (GlcNAc). In Serratia sp. ATCC 39006, NagC represses the transcription of two divergent operons, nagE and nagBAC, which encode proteins involved in the transport and metabolism of GlcNAc. Moreover, NagC directly binds to a 21-nt region that partially overlaps the -10 and -35 regions of the pig promoter and promotes the transcription of prodigiosin biosynthesis genes, thereby increasing prodigiosin production. Although NagC still acts as both repressor and activator in Serratia sp. ATCC 39006, its transcriptional regulatory activity is independent of GlcNAc. NagC was first found to regulate antibiotic biosynthesis in Gram-negative bacteria, and NagC-mediated regulation is not responsive to GlcNAc, which contributes to future studies on the regulation of secondary metabolism by NagC in other bacteria. IMPORTANCE: The ROK family transcription factor NagC is an important global regulator in the γ-proteobacteria. A large number of genes involved in the transport and metabolism of sugars, as well as those associated with biofilm formation and pathogenicity, are regulated by NagC. In all of these regulations, the transcriptional regulatory activity of NagC responds to the supply of GlcNAc in the environment. Here, we found for the first time that NagC can regulate antibiotic biosynthesis, whose transcriptional regulatory activity is independent of GlcNAc. This suggests that NagC may respond to more signals and regulate more physiological processes in Gram-negative bacteria.

Functional studies on tandem carbohydrate-binding modules of a multimodular enzyme possessing two catalytic domains.

Although functional studies on carbohydrate-binding module (CBM) have been carried out extensively, the role of tandem CBMs in the enzyme containing multiple catalytic domains (CDs) is unclear. Here, we identified a multidomain enzyme (Lc25986) with a novel modular structure from lignocellulolytic bacterial consortium. It consists of a mannanase domain, two CBM65 domains (LcCBM65-1/LcCBM65-2), and an esterase domain. To investigate CBM function and domain interactions, full-length Lc25986 and its variants were constructed and used for enzymatic activity, binding, and bioinformatic analyses. The results showed that LcCBM65-1 and LcCBM65-2 both bind mannan and xyloglucan but not cellulose or ß-1,3-1,4-glucan, which differs from the ligand specificity of reported CBM65s. Compared to LcCBM65-2, LcCBM65-1 showed a stronger ligand affinity and a preference for acetylation sites. Both CBM65s stimulated the enzymatic activities of their respective neighboring CDs against acetylated mannan, but did not contribute to the activities of the distal CDs. The time course of mannan hydrolysis indicated that the full-length Lc25986 was more effective in the complete degradation of mixed acetyl/non-acetyl substrates than the mixture of single-CD mutants. When acting on complex substrates, LcCBM65-1 not only improved the enzymatic activity of the mannanase domain, but also directed the esterase domain to the acetylated polysaccharides. LcCBM65-2 adopted a low affinity to reduce interference with the catalysis of the mannanase domain. These results demonstrate the importance of CBMs for the synergism between the two CDs of a multidomain enzyme and suggest that they contribute to the adequate degradation of complex substrates such as plant cell walls. IMPORTANCE: Lignocellulolytic enzymes, particularly those of bacterial origin, often harbor multiple carbohydrate-binding modules (CBMs). However, the function of CBM multivalency remains poorly understood. This is especially true for enzymes that contain more than one catalytic domain (CD), as the interactions between CDs, CBMs, and CDs and CBMs can be complex. Our research demonstrates that homogeneous CBMs can have distinct functions in a multimodular enzyme. The tandem CBMs coordinate the CDs in catalytic conflict through their differences in binding affinity, ligand preference, and arrangement within the full-length enzyme. Additionally, although the synergism between mannanase and esterase is widely acknowledged, our study highlights the benefits of integrating the two enzymes into a single entity for the degradation of complex substrates. In summary, these findings enhance our understanding of the intra-synergism of a multimodular enzyme and emphasize the significance of multiple CBMs in this context.

Multiresidue Determination of 26 Quinolones in Poultry Feathers Using UPLC-MS/MS and Their Application in Residue Monitoring.

As a non-traditional sample matrix, feather samples can be used to effectively monitor antibiotic addition and organismal residue levels in poultry feeding. Therefore, an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to simultaneously determine the residue levels of 26 quinolones in poultry feathers. The feather samples were extracted by sonication with a 1% formic acid and acetonitrile mixture in a water bath at 50 °C for 30 min, purified by the adsorption of multiple matrix impurities, dried with nitrogen, redissolved, and analyzed by UPLC-MS/MS. The linearity, limit of detection (LOD), limit of quantification (LOQ), recovery and precision were calculated. The 26 antibiotics demonstrated good linearity in the linear range. The recoveries and coefficients of variation were 78.9-110% and <13.7% at standard spiked levels of 10, 100 and 200 µg/kg, respectively. The LOD and LOQ were 0.12-1.31 and 0.96-2.60 µg/kg, respectively. The method also successfully identified quinolone residues in 50 poultry feather samples. The results showed that quinolones can accumulate and stabilize for a certain period of time after transferring from the body to the feathers of poultry.

One-step fermentation for producing xylo-oligosaccharides from wheat bran by recombinant Escherichia coli containing an alkaline xylanase.

BACKGROUND: One-step fermentation is a cheap way to produce xylo-oligosaccharides (XOS), where production of xylanases and XOS is integrated into a single process. In spite of cost advantage, one-step fermentation is still short in yield so far due to the limited exploration. To cope with this issue, production of XOS from wheat bran by recombinant Escherichia coli through one-step fermentation was investigated in this study. RESULTS: An endo-ß-1,4-xylanase gene belonging to glycoside hydrolase family 11 of Bacillus agaradhaerens was employed to construct recombinant E. coli. This xylanase showed maximal activity at 60 °C and pH 8.0-8.5. Its activity retained more than 60% after incubation at 70 °C for 4 h, showing a good stability. The recombinant E. coli BL21(DE3) could secreted xylanases that directly hydrolyzed de-starched wheat bran to XOS in fermentation medium. The XOS generated from hydrolysis consisted of xylose, xylobiose and xylotriose accounting for 23.1%, 37.3% and 39.6%, respectively. Wheat bran concentration was found to be the most crucial factor affecting XOS production. The XOS concentration reached 5.3 mg/mL at 10% loading of wheat bran, which is higher than those of previous researches. Nitrogen source type could also affect production of XOS by changing extracellular xylanase activity, and glycine was found to be the best one for fermentation. Optimal fermentation conditions were finally studied using response surface optimization. The maximal concentration emerged at 44.3 °C, pH 7.98, which is affected by characteristics of the xylanase as well as growth conditions of E. coli. CONCLUSIONS: This work indicates that the integrated fermentation using recombinant E. coli is highly competitive in cost and final concentration for producing XOS. Results can also provide theoretical basis for large-scale production and contribute to the wide adoption of XOS.

Organic Hydroperoxide Induces Prodigiosin Biosynthesis in Serratia sp. ATCC 39006 in an OhrR-Dependent Manner.

The biosynthesis of prodigiosin in the model prodigiosin-producing strain, Serratia sp. ATCC 39006, is significantly influenced by environmental and cellular signals. However, a comprehensive regulatory mechanism for this process has not been well established. In the present study, we demonstrate that organic hydroperoxide activates prodigiosin biosynthesis in an OhrR-dependent manner. Specifically, the MarR-family transcriptional repressor OhrR (Ser39006_RS05455) binds to its operator located far upstream of the promoter region of the prodigiosin biosynthesis operon (319 to 286 nucleotides [nt] upstream of the transcription start site) and negatively regulates the expression of prodigiosin biosynthesis genes. Organic hydroperoxide disassociates the binding between OhrR and its operator, thereby promoting the prodigiosin production. Moreover, OhrR modulates the resistance of Serratia sp. ATCC 39006 to organic hydroperoxide by regulating the transcription of its own gene and the downstream cotranscribed ohr gene. These results demonstrate that OhrR is a pleiotropic repressor that modulates the prodigiosin production and the resistance of Serratia sp. ATCC 39006 to organic hydroperoxide stress. IMPORTANCE Bacteria naturally encounter various environmental and cellular stresses. Organic hydroperoxides generated from the oxidation of polyunsaturated fatty acids are widely distributed and usually cause lethal oxidative stress by damaging cellular components. OhrR is known as a regulator that modulates the resistance of bacteria to organic hydroperoxide stress. In the current study, organic hydroperoxide disassociates OhrR from the promoter of prodigiosin biosynthesis gene cluster, thus promoting transcription of pigA to -O genes. In this model, organic hydroperoxide acts as an inducer of prodigiosin synthesis in Serratia sp. ATCC 39006. These results improve our understanding of the regulatory network of prodigiosin synthesis and serve as an example for identifying the cross talk between the stress responses and the regulation of secondary metabolism.

Threshold of main pancreatic duct for malignancy in intraductal papillary mucinous neoplasm at head-neck and body-tail.

BACKGROUND: Main pancreatic duct (MPD) dilation is a high-risk stigmata/worrisome feature of malignancy in intraductal papillary mucinous neoplasms (IPMNs). The threshold of MPD diameter in predicting malignancy may be related to the lesion location. This study aimed to separately identify the thresholds of MPD for malignancy of IPMNs separately for the head-neck and body-tail. MATERIALS AND METHODS: A total of 185 patients with pathologically confirmed IPMNs were included. Patient demographic information, clinical data, and pathological features were obtained from the medical records. Those IPMNs with high-grade dysplasia or with associated invasive carcinoma were considered as malignant tumor. Radiological data including lesion location, tumor size, diameter of the MPD, mural nodule, and IPMN types (main duct, MD; branch duct, BD; and mixed type, MT), were collected on computed tomography or magnetic resonance imaging. Serum carbohydrate antigen 19-9 levels, serum carcinoembryonic antigen levels, and the medical history of diabetes mellitus, chronic cholecystitis, and pancreatitis were also collected. RESULTS: Malignant IPMNs were detected in 31.6% of 117 patients with lesions in the pancreatic head-neck and 20.9% of 67 patients with lesions in the pancreatic body-tail. In MPD-involved IPMNs, malignancy was observed in 54.1% of patients with lesions in the pancreatic head-neck and 30.8% of patients with lesions in the pancreatic body-tail (p < 0.05). The cutoff value of MPD diameter for malignancy was 6.5 mm for lesions in the head-neck and 7.7 mm for lesions in the body-tail in all type of IPMNs. In MPD-involved IPMNs, the threshold was 8.2 mm for lesion in pancreatic head-neck and 7.7 mm for lesions in the body-tail. Multivariate analysis confirmed that MPD diameter ≥ 6.5 mm (pancreatic head-neck) and MPD diameter ≥ 7.7 mm (pancreatic body-tail) were independent predictors of malignancy (p < 0.05). Similar results were observed in MPD-involved IPMNs using 8.2 mm as a threshold. CONCLUSION: The thresholds of the dilated MPD may be associated with IPMNs locations. Thresholds of 6.5 mm for lesions in the head-neck and 7.7 mm for lesions in the body-tail were observed. For MPD-involved IPMNs alone, threshold for lesions in the head-neck was close to that in the body-tail.

Patients' choice and preference for common disease diagnosis and diabetes care: A discrete choice experiment.

OBJECTIVE: To examine the impact of policy incentives on patient's choice of health-care providers for primary care and to capture the preferences for general practitioner (GP) care. STUDY DESIGN: Discrete choice experiment. METHODS: A random sample of 704 adults and of 181 diabetics were independently surveyed to elicit patients' preferences for common disease diagnosis and diabetes care. Mixed logit regression was used for the analysis. RESULTS: On average, the most valued attribute in GP care are the organizational factors related to whether the provider has sufficient medicine and equipment to provide capable primary care service. Policy incentives, such as reducing waiting time, providing prior expert access, and increasing Medicare reimbursement, can facilitate the utilization of the GP system. Significant preference heterogeneity was identified; specifically, patient preferences significantly differ with regard to demand for common disease diagnosis and diabetes care. CONCLUSION: The identification of the preferences of specific groups in regard to GP care is an organizational and political imperative. Policy incentives are useful tools to guide patients' health care seeking behavior. To change the perceptions of Chinese patients with regard to health care, policy makers should consider the heterogeneous responses of residents to policy incentives and focus their efforts on key cohorts.

Decolorization and detoxification of water-insoluble Sudan dye by Shewanella putrefaciens CN32 co-cultured with Bacillus circulans BWL1061.

Effluents loaded with various synthetic dyes are considered as a huge burden to the surrounding ecosystems. Sudan dyes are relatively difficult to decolorize due to its water-insolubility. In the present study, the strain Shewanella putrefaciens CN32 was firstly applied to decolorize Sudan dyes under the anaerobic condition, and the physicochemical parameters on the decolorization were optimized. The results demonstrated that the suitable decolorization condition was temperature 26 °C, initial pH 7.0-8.0 and NaCl concentrations 0-20 g/L. Electron competitive acceptors including nitrite, nitrate, dimethyl sulphoxide and oxygen could cause the significant inhibition to the decolorization of Sudan dyes. Biosurfactant rhamnolipid played a positive role in enhancing the decolorization of Sudan I. The co-culture of S. putrefaciens CN32 and Bacillus circulans BWL1061 is reported for the first time to accelerate the decolorization through improving the synergistic effect of enzymatic degradation and biological reductive effect. The highest decolorization of 90.23% to Sudan I was achieved within 108 h, suggesting that co-culture technique has a good potential in the treatment of dyeing wastewater. Furthermore, the microbial toxicity tests indicated that the toxicity of Sudan I to Escherichia coli BL21 and Bacillus subtilis 168 was obviously decreased after the decolorization.

Use of aspirin in cardiovascular disease prevention-the role of online information search.

OBJECTIVES: This study intends to examine influences of online information search on the use of aspirin in cardiovascular diseases (CVDs) prevention among the applicable adult population in the United States. METHODS: We used data of 2018 National Health Interview Survey (NHIS). Our study sample is limited to adults age 40 or older to be consistent with the American Heart Association/American College of Cardiology Foundation (AHA/ACCF) guidelines for aspirin use. Linear probability models were used to test the association between patient's aspirin use behaviors and the variables of interest in four separate models. RESULTS: Our results show that the use of aspirin for CVD prevention was associated with online health information seeking in different ways. When patients received doctors' advice to use aspirin, online information seeking has a negative influence, depending on whether the individual has CVD risk factors. However, for patients without recommendations from providers, the effects of online information seeking on self-initiated aspirin use depend on the different types of preventions (primary vs. secondary) and CVD risk factors. CONCLUSION: Overall, online health information might lead to both overuse and underuse of aspirin in CVD preventions. Findings in this study may lead to decision-making that is not consistent with advice from healthcare professionals and/or established clinical guidelines.

High spatiotemporal-resolution mapping for a seasonal erosion flooding inundation using time-series Landsat and MODIS images.

Seasonal erosion flooding events present a significant challenge for effective disaster monitoring and land degradation studies. This research addresses this challenge by harnessing the combined capabilities of time-series Landsat and MODIS images to achieve high spatiotemporal-resolution mapping of flooding during such events. The study underscores the critical importance of precise flood monitoring for disaster mitigation and informed land management. To overcome the limitations posed by the trade-off between spatial and temporal resolution in current satellite sensors, we emplyedand theflexible spatiotemporal data fusion (FSDAF) methods to produce synthetic flood images with enhanced spatiotemporal resolutions for mapping by using MODIS and Landsat data from August 29 to September 3, 2016. A comparison was made between flood maps from several post-disaster forecasts based on ground-obtained time-series images of the Tumen River flood in China. According to the FSDAF approach, the input Landsat image of March 25, 2016, and the fused results had a root mean square error (RMSE) of 0.0301, average difference of 0.001, r of 0.941, and structure similarity indexof 0.939, indicating that temporal variation data had been effectively incorporated into a forecast on August 16, 2016. Results also indicated that the FSDAF forecast values are lower than those from the actual Landsat image. The results of the study also showed that the generated images could be effectively used for flood mapping. By using our newly developed simulation model, we were able to produce a comprehensive map of the inundated areas during the event from August 29 to September 3, 2016. This shows that FSDAF holds great potential for flood prediction and study and has the potential to benefit further disaster-related land degradation by combining multi-source images to provide high temporal and spatial resolution remote sensing information.

The global regulation of c-di-GMP and cAMP in bacteria.

Nucleotide second messengers are highly versatile signaling molecules that regulate a variety of key biological processes in bacteria. The best-studied examples are cyclic AMP (cAMP) and bis-(3'-5')-cyclic dimeric guanosine monophosphate (c-di-GMP), which both act as global regulators. Global regulatory frameworks of c-di-GMP and cAMP in bacteria show several parallels but also significant variances. In this review, we illustrate the global regulatory models of the two nucleotide second messengers, compare the different regulatory frameworks between c-di-GMP and cAMP, and discuss the mechanisms and physiological significance of cross-regulation between c-di-GMP and cAMP. c-di-GMP responds to numerous signals dependent on a great number of metabolic enzymes, and it regulates various signal transduction pathways through its huge number of effectors with varying activities. In contrast, due to the limited quantity, the cAMP metabolic enzymes and its major effector are regulated at different levels by diverse signals. cAMP performs its global regulatory function primarily by controlling the transcription of a large number of genes via cAMP receptor protein (CRP) in most bacteria. This review can help us understand how bacteria use the two typical nucleotide second messengers to effectively coordinate and integrate various physiological processes, providing theoretical guidelines for future research.

Carbohydrate-binding modules of ChiB and ChiC promote the chitinolytic system of Serratia marcescens BWL1001.

Carbohydrate-binding modules (CBMs) are commonly found within chitinases, but their contributions to chitinolytic systems are poorly understood. To address this knowledge gap, full-length chitin-acting enzymes (ChiA, ChiB, ChiC, and CBP21) of Serratia marcescens BWL1001 and CBM-truncated versions (ChiB-dCBM and ChiC-dCBM) were heterologously expressed for enzymological analysis. The CBM5 of ChiB and the CBM12 of ChiC both exhibited an affinity for α-chitin, while only CBM12 could bind colloidal chitin based on adsorption assays and affinity electrophoresis. Consistent with their ligand specificity, both CBMs were essential to α-chitin hydrolysis, while only CBM12 enhanced the hydrolytic efficiency of colloidal chitin by individual chitinases. Analysis of synergistic hydrolysis with separate full-length and CBM-truncated chitinases revealed that the two CBMs promoted the synergistic activity of chitinases on crystalline and amorphous chitin. The two CBMs also promoted the hydrolysis when chitin was mixed with non-substrate polysaccharides. This study reveals not only the effects of CBMs on enzymatic characteristics of individual chitinases, but also their contributions to the overall efficiency of chitinolytic systems during synergistic hydrolysis.

Functional identification of two novel carbohydrate-binding modules of glucuronoxylanase CrXyl30 and their contribution to the lignocellulose saccharification.

BACKGROUND: Glycoside hydrolase (GH) family 30 xylanases are a distinct group of xylanases, most of which have a highly specific catalytic activity for glucuronoxylan. Since GH30 xylanases do not normally carry carbohydrate-binding modules (CBMs), our knowledge of the function of their CBMs is lacking. RESULTS: In this work, the CBM functions of CrXyl30 were investigated. CrXyl30 was a GH30 glucuronoxylanase containing tandem CBM13 (CrCBM13) and CBM2 (CrCBM2) at its C terminus, which was identified in a lignocellulolytic bacterial consortium previously. Both CBMs could bind insoluble and soluble xylan, with CrCBM13 having binding specificity for the xylan with L-arabinosyl substitutions, whereas CrCBM2 targeted L-arabinosyl side chains themselves. Such binding abilities of these two CBMs were completely different from other CBMs in their respective families. Phylogenetic analysis also suggested that both CrCBM13 and CrCBM2 belong to novel branches. Inspection of the simulated structure of CrCBM13 identified a pocket that just accommodates the side chain of 3(2)-alpha-L-arabinofuranosyl-xylotriose, which forms hydrogen bonds with three of the five amino acid residues involved in ligand interaction. The truncation of either CrCBM13 or CrCBM2 did not alter the substrate specificity and optimal reaction conditions of CrXyl30, whereas truncation of CrCBM2 decreased the kcat/Km value by 83% (± 0%). Moreover, the absence of CrCBM2 and CrCBM13 resulted in a 5% (± 1%) and a 7% (± 0%) decrease, respectively, in the amount of reducing sugar released by the synergistic hydrolysis of delignified corncob whose hemicellulose is arabinoglucuronoxylan, respectively. In addition, fusion of CrCBM2 with a GH10 xylanase enhanced its catalytic activity against the branched xylan and improved the synergistic hydrolysis efficiency by more than fivefold when delignified corncob was used as substrate. Such a strong stimulation of hydrolysis resulted from the enhancement of hemicellulose hydrolysis on the one hand, and the cellulose hydrolysis is also improved according to the lignocellulose conversion rate measured by HPLC. CONCLUSIONS: This study identifies the functions of two novel CBMs in CrXyl30 and shows the good potential of such CBMs specific for branched ligands in the development of efficient enzyme preparations.

Gene expression profiling in the skin of zebrafish infected with Citrobacter freundii.

Skin is considered the largest immunologically active organ, but its molecular mechanism remains unclear in fish. Here, Affymetrix Zebrafish GeneChip was used to assess gene expression in the skin of zebrafish (Danio rerio) infected with the bacterium Citrobacter freundii. The results showed that 229 genes were differentially expressed, of which 196 genes were upregulated and 33 genes were downregulated. Gene Ontology and KEGG pathway analyses indicated 88 genes significantly associated with skin immunity involved in complement activation and acute phase response, defense and immune response, response to stress and stimulus, antigen processing and presentation, cell adhesion and migration, platelet activation and coagulation factors, regulation of autophagy and apoptosis. When compared with transcriptional profiles of previously reported carp (Cyprinus carpio) skin, a similar innate immunity (e.g., interferon, lectin, heat shock proteins, complements), and several different acute phase proteins (transferrin, ceruloplasmin, vitellogenin and alpha-1-microglobulin, etc.) were detected in zebrafish skin. The validity of the microarray results was verified by quantitative real-time PCR analysis of nine representative genes. This is first report that skin play important roles in innate immune responses to bacterial infection, which contribute to understanding the defense mechanisms of the skin in fish.

Willingness to use community health centres for initial diagnosis: the role of policy incentives among Chinese patients.

The aim of the study is to investigate the effect of policy incentives on residents' willingness to use community health centres for initial diagnosis. A cross-sectional survey with specific multiple price-list experiments was conducted in Guizhou, China. We were able to use in-depth individual interviews with a total of 422 participants. Our results showed that both financial and non-financial policy incentives can facilitate the utilisation of the community health centres. Approximately 60% of the respondents reported an increase in their willingness to use community health centres after the presentation of the financial policy, whereas 50% of respondents expressed an increase in their willingness to use community health centres with the non-financial policy. However, to some specific subgroups, such as residents with low trust, residents without chronic disease, residents with less healthcare visits and risk-averters, the impact of policy incentives were limited. The policy incentives are useful tools to attract more visitors to community health centres for initial diagnosis; however, their incentive effects vary in different subgroups. Thus, to change patients' perceptions regarding healthcare provider choice for initial diagnosis, policymakers should consider the heterogeneous responses of patients to policy incentives and focus their efforts on key cohorts.

Corrigendum to: Willingness to use community health centres for initial diagnosis: the role of policy incentives among Chinese patients.

The aim of the study is to investigate the effect of policy incentives on residents' willingness to use community health centres for initial diagnosis. A cross-sectional survey with specific multiple price-list experiments was conducted in Guizhou, China. We were able to use in-depth individual interviews with a total of 422 participants. Our results showed that both financial and non-financial policy incentives can facilitate the utilisation of the community health centres. Approximately 60% of the respondents reported an increase in their willingness to use community health centres after the presentation of the financial policy, whereas 50% of respondents expressed an increase in their willingness to use community health centres with the non-financial policy. However, to some specific subgroups, such as residents with low trust, residents without chronic disease, residents with less healthcare visits and risk-averters, the impact of policy incentives were limited. The policy incentives are useful tools to attract more visitors to community health centres for initial diagnosis; however, their incentive effects vary in different subgroups. Thus, to change patients' perceptions regarding healthcare provider choice for initial diagnosis, policymakers should consider the heterogeneous responses of patients to policy incentives and focus their efforts on key cohorts.

cAMP and c-di-GMP synergistically support biofilm maintenance through the direct interaction of their effectors.

Nucleotide second messengers, such as cAMP and c-di-GMP, regulate many physiological processes in bacteria, including biofilm formation. There is evidence of cross-talk between pathways mediated by c-di-GMP and those mediated by the cAMP receptor protein (CRP), but the mechanisms are often unclear. Here, we show that cAMP-CRP modulates biofilm maintenance in Shewanella putrefaciens not only via its known effects on gene transcription, but also through direct interaction with a putative c-di-GMP effector on the inner membrane, BpfD. Binding of cAMP-CRP to BpfD enhances the known interaction of BpfD with protease BpfG, which prevents proteolytic processing and release of a cell surface-associated adhesin, BpfA, thus contributing to biofilm maintenance. Our results provide evidence of cross-talk between cAMP and c-di-GMP pathways through direct interaction of their effectors, and indicate that cAMP-CRP can play regulatory roles at the post-translational level.

Dynamic changes in community structure and degradation performance of a bacterial consortium MMBC-1 during the subculturing revival reveal the potential decomposers of lignocellulose.

Bacterial consortium is an important source of lignocellulolytic strains, but it is still a challenge to distinguish the direct decomposers of lignocellulose from other bacteria in such a complex community. This study aims at addressing this issue by focusing on the dynamic changes in community structure and degradation activity of MMBC-1, an established and stable lignocellulolytic bacterial consortium, during its subculturing revival. MMBC-1 was cryopreserved with glycerol as a protective agent and then inoculated for revival. Its enzyme activities for degradation recovered to the maximum level after two rounds of subculturing. Correspondingly, the cellulose and hemicellulose in lignocellulosic carbon source were gradually decomposed during the revival. Meanwhile, the initial dominant bacteria represented by genus Clostridium were replaced by the bacteria belonging to Lachnospira, Enterococcus, Bacillus, Haloimpatiens genera and family Lachnospiraceae. However, only three high-abundance (> 1%) operational taxonomic units (OTUs) (Lachnospira, Enterococcus and Haloimpatiens genera) were suggested to directly engage in lignocellulose degradation according to correlation analysis. By comparison, many low-abundance OTUs, such as the ones belonging to Flavonifractor and Anaerotruncus genera, may play an important role in degradation. These findings showed the dramatic changes in community structure that occurred during the subculturing revival, and paved the way for the discovery of direct decomposers in a stable consortium.

Aging Resistance of Silica Fume/Styrene-Butadiene-Styrene Composite-Modified Asphalt.

The influences of silica fume content and aging on the rheological properties of silica fume/styrene-butadiene-styrene composite-modified asphalts were investigated via rolling thin-film oven test simulations. The asphalts rheological properties before and after aging were measured using three-major-indices, dynamic shear rheology, and bending beam rheometer tests. Fourier transform infrared spectroscopy was used to examine the changes in the functional groups of the asphalt. The silica fume did not chemically react with the modified asphalt, and its original structure was maintained. The aging resistance improved significantly after adding the silica fume. At 6% silica fume content, the relaxation of the asphalt was the highest, indicating that the asphalt had the best low-temperature crack resistance at this mixing proportion. Furthermore, the carbonyl index value of this sample exhibited the smallest increment among all of the samples, and this asphalt sample had the strongest short-term aging resistance. Thus, the optimum silica fume content in the composite-modified asphalt was determined to be 6%. This information may be used to fabricate an asphalt mixture that can improve the service life and aging resistance of pavements.

Recent advances and perspectives in efforts to reduce the production and application cost of microbial flocculants.

Microbial flocculants are macromolecular substances produced by microorganisms. Due to its non-toxic, harmless, and biodegradable advantages, microbial flocculants have been widely used in various industrial fields, such as wastewater treatment, microalgae harvest, activated sludge dewatering, heavy metal ion adsorption, and nanoparticle synthesis, especially in the post-treatment process of fermentation with high safety requirement. However, compared with the traditional inorganic flocculants and organic polymeric flocculants, the high production cost is the main bottleneck that restricts the large-scale production and application of microbial flocculants. To reduce the production cost of microbial flocculant, a series of efforts have been carried out and some exciting research progresses have been achieved. This paper summarized the research advances in the last decade, including the screening of high-yield strains and the construction of genetically engineered strains, search of cheap alternative medium, the extraction and preservation methods, microbial flocculants production as an incidental product of other biological processes, combined use of traditional flocculant and microbial flocculant, and the production of microbial flocculant promoted by inducer. Moreover, this paper prospects the future research directions to further reduce the production cost of microbial flocculants, thereby promoting the industrial production and large-scale application of microbial flocculants.