Development of an instrument to measure the competencies of health professionals in the process of evidence-based healthcare: A Delphi study.

AIMS: To identify and reach consensus on dimensions and criteria of a competence assessment instrument for health professionals in relation to the process of evidence-based healthcare. DESIGN: A two-round Delphi survey was carried out from April to June 2023. METHODS: Consensus was sought from an expert panel on the instrument preliminarily established based on the JBI Model of Evidence-Based Healthcare and a rapid review of systematic reviews of relevant literature. The level of consensus was reflected by the concentration and coordination of experts' opinions and percentage of agreement. The instrument was revised significantly based on the combination of data analysis, the experts' comments and research group discussions. RESULTS: Sixteen national and three international experts were involved in the first-round Delphi survey and 17 experts participated in the second-round survey. In both rounds, full consensus was reached on the four dimensions of the instrument, namely evidence-generation, evidence-synthesis, evidence-transfer and evidence-implementation. In round-one, the instrument was revised from 77 to 61 items. In round-two, the instrument was further revised to have 57 items under the four dimensions in the final version. CONCLUSION: The Delphi survey achieved consensus on the instrument. The validity and reliability of the instrument needs to be tested in future research internationally. IMPLICATIONS FOR THE PROFESSION AND/OR PATIENT CARE: Systematic assessment of nurses and other health professionals' competencies in different phases of evidence-based healthcare process based on this instrument provides implications for their professional development and multidisciplinary team collaboration in evidence-based practice and better care process and outcomes. IMPACT: This study addresses a research gap of lacking an instrument to systematically assess interprofessional competencies in relation to the process of EBHC. The instrument covers the four phases of EBHC process with minimal criteria, highlighting essential aspects of ability to be developed. Identification of health professionals' level of competence in these aspects helps strengthen their capacity accordingly so as to promote virtuous EBHC ecosystem for the ending purpose of improving global healthcare outcomes. REPORTING METHOD: This study was reported in line with the Conducting and REporting of DElphi studies (CREDES) guidance on Delphi studies. PATIENT AND PUBLIC CONTRIBUTION: No patient or public contribution.

Profiling and source tracking of the microbial populations and resistome present in fish products.

Microorganisms in processing environments significantly impact the quality and safety of food products and can serve as potential reservoirs for antibiotic-resistant genes, contributing to public health concerns about antimicrobial resistance (AMR). Fish processing plants represent an understudied environment for microbiome mapping. This study investigated the microbial composition, prevalence of Listeria spp., and resistome structures in three catfish processing facilities in the southeastern United States. The 16S rRNA gene sequencing revealed that the observed richness and Shannon diversity index increased significantly from fish to fillet. Beta diversity analysis showed distinct clustering of microbial communities between fish, environment, and fillet samples. Fast expectation-maximization microbial source tracking (FEAST) algorithm demonstrated that the microbiota presents in the processing environment contributed 48.2 %, 62.4 %, and 53.7 % to the microbiota present on fillet in Facility 1 (F1), F2, and F3, respectively. Food contact surfaces made larger contributions compared to the non-food contact surfaces. The linear discriminant analysis of effect size (LEfSe) identified specific microbial genera (e.g., Plesiomohas, Brochothrix, Chryseobacterium and Cetobacterium) that significantly varied between Listeria spp. positive and negative samples in all three processing plants. The metagenomic sequencing results identified 212 antimicrobial resistance genes (ARGs) belonging to 72 groups from the raw fish and fish fillet samples collected from three processing plants. Although there was a significant decrease in the overall diversity of ARGs from fish to fillet samples, the total abundance of ARGs did not change significantly (P > 0.05). ARGs associated with resistance to macrolide-lincosamide-streptogramin (MLS), cationic antimicrobial peptides, aminoglycosides, and beta-lactams were found to be enriched in the fillet samples when compared to fish samples. Results of this study highlight the profound impact of processing environment on shaping the microbial populations present on the final fish product and the need for additional strategies to mitigate AMR in fish products.

Reduction Foodborne Pathogens and Surrogate Microorganism on Citrus Fruits after Lab- and Pilot-scale Finishing Wax Application.

After finishing waxes are applied, citrus fruits are typically dried at 32-60°C for 2-3 min before final packing. The survival of Listeria monocytogenes, Salmonella, and Enterococcus faecium NRRL B-2354 was evaluated under laboratory conditions on lemons after applying one of four finishing waxes (F4, F6, F8, and F15) followed by an ambient hold or heated (50 or 60°C) drying step. The reduction of inoculated microorganisms during drying was significantly influenced by wax type and temperature, with greater reductions at higher temperatures. Greater reductions after waxing and drying at 60°C were observed with L. monocytogenes (2.84-4.44 log) than with Salmonella (1.65-3.67 log), and with Salmonella than with E. faecium (0.99-2.93 log). The survival of Salmonella inoculated at 5.8-5.9 log/fruit on lemons and oranges after applying wax F6 and drying at 60°C was evaluated during storage at 4 and 22°C. The reductions of Salmonella after waxing and drying were 1.7 log; additional reductions during storage at 4 or 22°C were 1.40-1.43 or 0.18-0.29 log, respectively, on waxed lemons, and 0.56-1.02 or 0.54-0.57 log, respectively, on waxed oranges. Under pilot-scale packinghouse conditions with wax F4, mean and minimum reductions of E. faecium ranged from 2.15 to 2.89 and 1.64 to 2.12 log, respectively. However, E. faecium was recovered by whole-fruit enrichment (limit of detection: 0.60 log CFU/lemon) but not by plating (LOD: 1.3 log CFU/lemon) from uninoculated lemons run with or after the inoculated lemons. The findings should provide useful information to establish and implement packinghouse food safety plans.

Design of Hollow Porous P-NiCo2O4@Co3O4 Nanoarray and Its Alkaline Aqueous Zinc-Ion Battery Performance.

Alkaline aqueous zinc-ion batteries possess a wider potential window than those in mildly acidic systems; they can achieve high energy density and are expected to become the next generation of energy storage devices. In this paper, a hollow porous P-NiCo2O4@Co3O4 nanoarray is obtained by ion etching and the calcination and phosphating of ZiF-67, which is directly grown on foam nickel substrate, as a precursor. It exhibits excellent performance as a cathode material for alkaline aqueous zinc-ion batteries. A high discharge specific capacity of 225.3 mAh g-1 is obtained at 1 A g-1 current density, and it remains 81.9% when the current density is increased to 10 A g-1. After one thousand cycles of charging and discharging at 3 A g-1 current density, the capacity retention rate is 88.8%. Even at an excellent power density of 25.5 kW kg-1, it maintains a high energy density of 304.5 Wh kg-1. It is a vital, promising high-power energy storage device for large-scale applications.

Warming increases the differences among spring phenology models under future climate change.

Phenological models are built upon an understanding of the influence of environmental factors on plant phenology, and serve as effective tools for predicting plant phenological changes. However, the differences in phenological model predictive performance under different climate change scenarios have been rarely studied. In this study, we parameterized thirteen spring phenology models, including six one-phase models and seven two-phase models, by combining phenological observations and meteorological data. Using climatic data from two Shared Socioeconomic Pathways (SSP) scenarios, namely SSP126 (high mitigation and low emission) and SSP585 (no mitigation and high emission), we predicted spring phenology in Germany from 2021 to 2100, and compared the impacts of dormancy phases and driving factors on model predictive performance. The results showed that the average correlation coefficient between the predicted start of growing season (SOS) by the 13 models and the observed values exceeded 0.72, with the highest reaching 0.80. All models outperformed the NULL model (Mean of SOS), and the M1 model (driven by photoperiod and forcing temperature) performed the best for all the tree species. In the SSP126 scenario, the average SOS advanced initially and then gradually shifted towards a delay starting around 2070. In the SSP585 scenario, the average SOS advanced gradually at a rate of approximately 0.14 days per year. Moreover, the standard deviation of the simulated SOS by the 13 spring phenology models exhibited a significant increase at a rate of 0.04 days per year. On average, two-phase models exhibited larger standard deviations than one-phase models after approximately 2050. Models driven solely by temperature showed larger standard deviations after 2060 compared to models driven by both temperature and photoperiod. Our findings suggest investigating the release mechanisms of endodormancy phase and incorporating new insights into future phenological models to better simulate the changes in plant phenology.

Formation of Listeria monocytogenes persister cells in the produce-processing environment.

Persisters are a subpopulation of growth-arrested cells that possess transient tolerance to lethal doses of antibiotics and can revert to an active state under the right conditions. Persister cells are considered as a public health concern. This study examined the formation of persisters by Listeria monocytogenes (LM) in an environment simulating a processing plant for leafy green production. Three LM strains isolated from California produce-processing plants and packinghouses with the strongest adherence abilities were used for this study. The impact of the cells' physiological status, density, and nutrient availability on the formation of persisters was also determined. Gentamicin at a dose of 100 mg/L was used for the isolation and screening of LM persisters. Results showed that the physiological status differences brought by culture preparation methods (plate-grown vs. broth-grown) did not impact persister formation (P > 0.05). Instead, higher persister ratios were found when cell density increased (P < 0.05). The formation of LM persister cells under simulated packinghouse conditions was tested by artificially inoculating stainless steel coupons with LM suspending in media with decreasing nutrient levels: brain heart infusion broth (1366 mg/L O2), produce-washing water with various organic loads (1332 mg/L O2 and 652 mg/L O2, respectively), as well as sterile Milli-Q water. LM survived in all suspensions at 4 °C with 85 % relative humidity for 7 days, with strain 483 producing the most persister cells (4.36 ± 0.294 Log CFU/coupon) on average. Although persister cell levels of LM 480 and 485 were reasonably steady in nutrient-rich media (i.e., BHI and HCOD), they declined in nutrient-poor media (i.e., LCOD and sterile Milli-Q water) over time. Persister populations decreased along with total viable cells, demonstrating the impact of available nutrients on the formation of persisters. The chlorine sensitivity of LM persister cells was evaluated and compared with regular LM cells. Results showed that, despite their increased tolerance to the antibiotic gentamicin, LM persisters were more susceptible to chlorine treatments (100 mg/L for 2 min) than regular cells.

Development and validation of a questionnaire to evaluate the knowledge, attitude, behaviour and care preference of family members of Chinese older adults related to palliative care.

AIM: To develop and validate a questionnaire on knowledge, attitude, behaviour and care preference of family members of Chinese older adults related to palliative care. DESIGN: A descriptive study design and STROBE checklist were applied in this research. METHODS: The theoretical framework of the questionnaire was knowledge-attitude-behaviour model. An additional dimension of palliative care preference of family members was set up in the questionnaire. Items were generated from a rapid review of international literature and interviews with 61 family members of the older adults living either in an aged care service organization or the community. The content validity was examined by five experts. A preliminary questionnaire with 69 items was then set up, and its psychometric property was assessed. RESULTS: A final version of questionnaire with 42 items under four dimensions was constructed. The content validity index of the overall questionnaire was 0.93 and of each item ranged 0.80-1.00. The factor loading of all items was higher than 0.50 as per exploratory and confirmatory factor analysis; the average variance extracted for each dimension was higher than 0.50; the composite reliability was higher than 0.90; and the absolute value of the correlation coefficient of each dimension was <0.50 and less than the square root of the average variance extracted. The Cronbach's alpha value and the split-half reliability value of the overall questionnaire were 0.93 and 0.97, respectively. CONCLUSIONS: This questionnaire has good validity and reliability, but needs further testing in multi-centered settings.

Consumption of common illicit drugs in twenty-one cities in southwest China through wastewater analysis.

Wastewater-based epidemiology (WBE) was applied to estimate illicit drugs consumption at a provincial scale in southwest China. A large-scale wastewater sampling campaign was carried out from October to November in 2021 in 156 different wastewater treatment plants (WWTPs). Two 24-h composite influent wastewater samples were collected in each WWTP. Concentrations of 11 illicit drugs or their metabolites were determined using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Benzoylecgonine, cocaine, 6-monoacetylmorphine, norketamine, 3,4-methylenedioxymethamphetamine (MDMA), and MDA were not detected in any of the wastewater samples. Methamphetamine and morphine were detected in >84% of samples, while ketamine was found in about 6% of the samples. The city-specific population-weighted consumption of methamphetamine and ketamine were in the range of 0.6-49.7 and N.D.-7.0 mg 1000 inh-1 day-1, respectively, with provincial population-weighted values of 22.6 and 2.4 mg 1000 inh-1 day-1 in southwest China. The city-specific load of morphine varied from 3.2 to 10.2 mg 1000 inh-1 day-1, with provincial population-weighted load of 6.7 mg 1000 inh-1 day-1. Taking into account therapeutic use of morphine and codeine, the provincial heroin consumption was estimated to be 10.3 mg 1000 inh-1 day-1, ranging from 1.7 to 18.5 mg 1000 inh-1 day-1 in 21 cities. Overall, the patterns of illicit drugs use were similar across southwest China, with high prevalence of methamphetamine and heroin, but relatively low use of ketamine. These findings could provide accurate drugs consumption information for timely identifying potential hotspots of illicit drugs use in southwest China.

Genome-Wide Identification and Transcriptional Expression Profiles of PP2C in the Barley (Hordeum vulgare L.) Pan-Genome.

The gene family protein phosphatase 2C (PP2C) is related to developmental processes and stress responses in plants. Barley (Hordeum vulgare L.) is a popular cereal crop that is primarily utilized for human consumption and nutrition. However, there is little knowledge regarding the PP2C gene family in barley. In this study, a total of 1635 PP2C genes were identified in 20 barley pan-genome accessions. Then, chromosome localization, physical and chemical feature predictions and subcellular localization were systematically analyzed. One wild barley accession (B1K-04-12) and one cultivated barley (Morex) were chosen as representatives to further analyze and compare the differences in HvPP2Cs between wild and cultivated barley. Phylogenetic analysis showed that these HvPP2Cs were divided into 12 subgroups. Additionally, gene structure, conserved domain and motif, gene duplication event detection, interaction networks and gene expression profiles were analyzed in accessions Morex and B1K-04-12. In addition, qRT-PCR experiments in Morex indicated that seven HvMorexPP2C genes were involved in the response to aluminum and low pH stresses. Finally, a series of positively selected homologous genes were identified between wild accession B1K-04-12 and another 14 cultivated materials, indicating that these genes are important during barley domestication. This work provides a global overview of the putative physiological and biological functions of PP2C genes in barley. We provide a broad framework for understanding the domestication- and evolutionary-induced changes in PP2C genes between wild and cultivated barley.

Resolving the Dust Bowl paradox of grassland responses to extreme drought.

During the 1930s Dust Bowl drought in the central United States, species with the C3 photosynthetic pathway expanded throughout C4-dominated grasslands. This widespread increase in C3 grasses during a decade of low rainfall and high temperatures is inconsistent with well-known traits of C3 vs. C4 pathways. Indeed, water use efficiency is generally lower, and photosynthesis is more sensitive to high temperatures in C3 than C4 species, consistent with the predominant distribution of C3 grasslands in cooler environments and at higher latitudes globally. We experimentally imposed extreme drought for 4 y in mixed C3/C4 grasslands in Kansas and Wyoming and, similar to Dust Bowl observations, also documented three- to fivefold increases in C3/C4 biomass ratios. To explain these paradoxical responses, we first analyzed long-term climate records to show that under nominal conditions in the central United States, C4 grasses dominate where precipitation and air temperature are strongly related (warmest months are wettest months). In contrast, C3 grasses flourish where precipitation inputs are less strongly coupled to warm temperatures. We then show that during extreme drought years, precipitation-temperature relationships weaken, and the proportion of precipitation falling during cooler months increases. This shift in precipitation seasonality provides a mechanism for C3 grasses to respond positively to multiyear drought, resolving the Dust Bowl paradox. Grasslands are globally important biomes and increasingly vulnerable to direct effects of climate extremes. Our findings highlight how extreme drought can indirectly alter precipitation seasonality and shift ecosystem phenology, affecting function in ways not predictable from key traits of C3 and C4 species.

[Efficacy of Autologous DC-CIK Cells Combined with Other Immune Cells in the Treatment of Hematological Malignancies -Retrospective Study].

OBJECTIVE: To investigate the safety and clinical efficacy of autologous DC-CIK cells combined with other immune cells for patients with hematological malignancies and analyze patient prognosis. METHODS: 50 patients with hematological malignancies who received cellular immunotherapy from September 2014 to April 2016 were retrospectively studied in the First Affiliated Hospital of Xi'an Jiaotong University, 115 cases times of cellular immunotherapy were performed. According to the selected treatment, the patients were divided into the dual cell group (DC-CIK cell treatment) and the multi-cell group (DC-CIK cell combined with other immune cells); According to the treatment course, the patients were divided into the single course group (completed by ＜3 times) and the multiple course group. The changes of T lymphocyte subsets, blood routine indicators and KPS scores as well as the overall survival time before and after treatment were compared and analyzed. RESULTS: [WTB1]The difference of general conditions before treatment including the number of patients, sex, age, T lymphocyte subsets, blood routine indicators, KPS scores and so on in 2 groups divided according to 2 kinds of treatment methods were not statistically significant, indicating that the 2 groups were comparable. Grouped by selected treatment, the CD4+/CD8+ ratio, Hb and Plt levels decreased in the dual cell group, compared with those before treatment(P＜0.05). The CD3+CD4+ ratio after treatment in multiple cell group decreased, compared with that before treatment (P＜0.05). The 3-year survival rate of patients in dual cell and multiple cell groups was 61.3% vs 69.8%, the overall survival time of patients in 2 groups was 32.4 months vs 39.6 months, there were no statisticall differences between 2 groups(P>0.05). Grouped by treatment course, the CD3+ ratio after treatment increased, while the Hb level after treatment decreased in single course group, compared with level before treatment(P＜0.05). The CD3+CD4+ ratio, Plt level decreased, while the KPS scores increased after treatment in multiple course group, compared with those before treatment(P＜0.05). The 3-year survival rate in single course and multiple course groups was 52% vs 76.4%, the overall survival time was 28.7 months vs 40.9 months respectively, statistically significant with difference (P＜0.05). CONCLUSION: Autologous DC-CIK cells combined with other immune cells in the treatment of hematological malignancies can change the immune function of the patients and improve the antitumor activity. The multi-course treatment can improve the quality of life, prolong the overall survival time, thus worthing clinical promotion.

Platinum-Ion-Mediated Self-Assembly of Hairpin Peptides and Synthesis of Platinum Nanostructures.

Nanostructures and nanomaterials based on peptide self-assembly have attracted tremendous interests due to the functionalities of peptide molecules. Furthermore, the self-assembled peptide nanostructures are also adopted to fabricate nanomaterials and nanodevices. In this work, the intramolecular folding and self-assembly of a ß-hairpin peptide CBHH were first studied under the regulation of platinum ion. And then, platinum nanostructures were synthesized through the reduction of platinum ions templated with peptide self-assemblies. The results of circular dichroism spectroscopy, UV-vis spectroscopy, isothermal titration calorimetry, and atomic force microscopy observation showed that platinum ions could promote the conversion of peptide CBHH secondary structure from a random coil to a ß-sheet through coordination with histidine residues. Platinum nanostructures including nanorods and one dimensionally aligned nanorods were synthesized through in situ reduction with CBHH self-assembled nanofiber as the templates. And the synthesized platinum nanostructures showed excellent electrocatalytic activities.

A general gelation strategy for 1D nanowires: dynamically stable functional gels for 3D printing flexible electronics.

3D printing of functional inorganic nanowires has been accomplished using extruded nanowire-based inks obtained by incorporating nanofillers into polymeric matrices or thickeners. However, the presence of inactive additives poses a critical challenge for fully realizing the functionality of the nanowires in the printed structures, which remains a fundamental hurdle to overcome. Here, to construct 3D-printed electronics with high performance, we developed a versatile gelation strategy to obtain thixotropic nanowire gels through formation of dynamically stable 3D networks using small amounts of flexible, water-soluble and single-layer 2D nanosheets, such as graphene oxide and MXene, as physical cross-linkers. The nanosheets can knot-tie and stabilize the nanowire junctions in the aqueous suspension, leading to the formation of stable and thixotropic gels with viscosities up to ∼80 000 Pa s at 0.01 s-1 in the absence of polymer thickener. Gels of varioius metallic and semiconductive nanowires have been successfully prepared and printed into 3D and self-supported architectures via extrusion-based 3D-printing. The synergism of nanowires and nanosheets not only conquers the restraints of harsh post-treatments to remove additives after printing, but also maximizes the functionality of the nanowires in the printed architectures. The printed 3D structures solidified by ambient drying, coagulation, or freeze-drying exhibit remarkable functionalities. For example, the electrical conductivity of the 3D-printed silver nanowire-based architectures can reach 40 000 S cm-1. The feasibility of these functional nanowire gels was demonstrated by fabricating a series of printed flexible electronics via extrusion-based 3D-printing.

High expression of type I inositol 1,4,5-trisphosphate receptor in the kidney of rats with hepatorenal syndrome.

AIM: To detect the expression of type I inositol 1,4,5-trisphosphate receptor (IP3RI) in the kidney of rats with hepatorenal syndrome (HRS). METHODS: One hundred and twenty-five Sprague-Dawley rats were randomly divided into four groups to receive an intravenous injection of D-galactosamine (D-GalN) plus lipopolysaccharide (LPS; group G/L, n = 50), D-GalN alone (group G, n = 25), LPS alone (group L, n = 25), and normal saline (group NS, n = 25), respectively. At 3, 6, 9, 12, and 24 h after injection, blood, liver, and kidney samples were collected. Hematoxylin-eosin staining of liver tissue was performed to assess hepatocyte necrosis. Electron microscopy was used to observe ultrastructural changes in the kidney. Western blot analysis and real-time PCR were performed to detect the expression of IP3RI protein and mRNA in the kidney, respectively. RESULTS: Hepatocyte necrosis was aggravated gradually, which was most significant at 12 h after treatment with D-galactosamine/lipopolysaccharide, and was characterized by massive hepatocyte necrosis. At the same time, serum levels of biochemical indicators including liver and kidney function indexes were all significantly changed. The structure of the renal glomerulus and tubules was normal at all time points. Western blot analysis indicated that IP3RI protein expression began to rise at 3 h (P < 0.05) and peaked at 12 h (P < 0.01). Real-time PCR demonstrated that IP3RI mRNA expression began to rise at 3 h (P < 0.05) and peaked at 9 h (P < 0.01). CONCLUSION: IP3RI protein expression is increased in the kidney of HRS rats, and may be regulated at the transcriptional level.

GC-MS Analysis of the Volatile Constituents in the Leaves of 14 Compositae Plants.

The green organs, especially the leaves, of many Compositae plants possess characteristic aromas. To exploit the utility value of these germplasm resources, the constituents, mainly volatile compounds, in the leaves of 14 scented plant materials were qualitatively and quantitatively compared via gas chromatography-mass spectrometry (GC-MS). A total of 213 constituents were detected and tentatively identified in the leaf extracts, and terpenoids (especially monoterpene and sesquiterpene derivatives), accounting for 40.45-90.38% of the total compounds, were the main components. The quantitative results revealed diverse concentrations and compositions of the chemical constituents between species. Principal component analysis (PCA) showed that different groups of these Compositae plants were characterized by main components of α-thujone, germacrene D, eucalyptol, ß-caryophyllene, and camphor, for example. On the other hand, cluster memberships corresponding to the molecular phylogenetic framework, were found by hierarchical cluster analysis (HCA) based on the terpenoid composition of the tested species. These results provide a phytochemical foundation for the use of these scented Compositae plants, and for the further study of the chemotaxonomy and differential metabolism of Compositae species.

Dynamic control of the mevalonate pathway expression for improved zeaxanthin production in Escherichia coli and comparative proteome analysis.

Engineered heterologous multi-gene metabolic pathways often suffer from flux imbalance and toxic metabolites, as the production host typically lacks the regulatory mechanisms for the heterologous pathway. Here, we first coordinated the expression of all genes of the mevalonate (MEV) pathway from Saccharomyces cerevisiae using the tunable intergenic regions (TIGRs), and then dynamically regulated the TIGR-mediated MEV pathway to prevent the accumulation of toxic metabolites by using IPP/FPP-responsive promoter. After introduction of the dynamically controlled TIGR-mediated MEV pathway into Escherichia coli, the content and concentration of zeaxanthin in shaker flask cultures were 2.0- and 2.1-fold higher, respectively, than those of the strain harboring the statically controlled non-TIGR-mediated MEV pathway. The content and concentration of zeaxanthin in E. coli ZEAX (pZSPgadE-MevTTIGR-MevBTIGRIS-2) reached 722.46mg/L and 23.16mg/g dry cell weight (DCW), respectively, in 5.0L fed-batch fermentation. We also comparatively analyzed the proteomes between E. coli ZEAX and E. coli ZEAX (pZSPgadE-MevTTIGR-MevBTIGRIS-2) to understand the mechanism of zeaxanthin biosynthesis. The results of the comparative proteomes demonstrate that zeaxanthin overproduction may be associated with increased precursor availability, increased NADPH availability, increased ATP availability, oxidative stress response, and increased membrane storage capacity for zeaxanthin due to changes in both cellular shape and membrane composition.

Temperature, precipitation, and insolation effects on autumn vegetation phenology in temperate China.

Autumn phenology plays a critical role in regulating climate-biosphere interactions. However, the climatic drivers of autumn phenology remain unclear. In this study, we applied four methods to estimate the date of the end of the growing season (EOS) across China's temperate biomes based on a 30-year normalized difference vegetation index (NDVI) dataset from Global Inventory Modeling and Mapping Studies (GIMMS). We investigated the relationships of EOS with temperature, precipitation sum, and insolation sum over the preseason periods by computing temporal partial correlation coefficients. The results showed that the EOS date was delayed in temperate China by an average rate at 0.12 ± 0.01 days per year over the time period of 1982-2011. EOS of dry grassland in Inner Mongolia was advanced. Temporal trends of EOS determined across the four methods were similar in sign, but different in magnitude. Consistent with previous studies, we observed positive correlations between temperature and EOS. Interestingly, the sum of precipitation and insolation during the preseason was also associated with EOS, but their effects were biome dependent. For the forest biomes, except for evergreen needle-leaf forests, the EOS dates were positively associated with insolation sum over the preseason, whereas for dry grassland, the precipitation over the preseason was more dominant. Our results confirmed the importance of temperature on phenological processes in autumn, and further suggested that both precipitation and insolation should be considered to improve the performance of autumn phenology models.

CmWRKY15 Facilitates Alternaria tenuissima Infection of Chrysanthemum.

Abscisic acid (ABA) has an important role in the responses of plants to pathogens due to its ability to induce stomatal closure and interact with salicylic acid (SA) and jasmonic acid (JA). WRKY transcription factors serve as antagonistic or synergistic regulators in the response of plants to a variety of pathogens. Here, we demonstrated that CmWRKY15, a group IIa WRKY family member, was not transcriptionally activated in yeast cells. Subcellular localization experiments in which onion epidermal cells were transiently transfected with CmWRKY15 indicated that CmWRKY15 localized to the nucleus in vivo. The expression of CmWRKY15 could be markedly induced by the presence of Alternaria tenuissima inoculum in chrysanthemum. Furthermore, the disease severity index (DSI) data of CmWRKY15-overexpressing plants indicated that CmWRKY15 overexpression enhanced the susceptibility of chrysanthemum to A. tenuissima infection compared to controls. To illustrate the mechanisms by which CmWRKY15 regulates the response to A. tenuissima inoculation, the expression levels of ABA-responsive and ABA signaling genes, such as ABF4, ABI4, ABI5, MYB2, RAB18, DREB1A, DREB2A, PYL2, PP2C, RCAR1, SnRK2.2, SnRK2.3, NCED3A, NCED3B, GTG1, AKT1, AKT2, KAT1, KAT2, and KC1were compared between transgenic plants and controls. In summary, our data suggest that CmWRKY15 might facilitate A. tenuissima infection by antagonistically regulating the expression of ABA-responsive genes and genes involved in ABA signaling, either directly or indirectly.

Engineering of Escherichia coli for Lycopene Production Through Promoter Engineering.

The control of gene expression is critical for metabolic engineering. The multi-copy plasmids has been widely used for high-level expression of genes. However, plasmid-based expression systems are liable to genetic instability and require a selective pressure to assure plasmid stability. In this study, we first constructed a lycopene producer Escherichia coli through promoter engineering. Saccharomyces cerevisiae mevalonate (MEV) pathway was also optimized to balance expression of the top and bottom MEV pathway by using the different strength promoters. The chromosomal heterologous expression of the optimized S. cerevisiae MEV pathway can further improved lycopene production. The final engineered strain, E. coli LYCOP 20, produced lycopene of 529.45 mg/L and 20.25 mg per gram of dry cell weight in the fed-batch culture. The engineered strain does not have a plasmid or antibiotic marker. This strategy used in this study can be applied in pathway engineering of E. coli and other bacteria.

Metabolic engineering of Escherichia coli to produce zeaxanthin.

Zeaxanthin is a high-value carotenoid that is used in nutraceuticals, cosmetics, food, and animal feed industries. Zeaxanthin is chemically synthesized or purified from microorganisms as a natural product; however, increasing demand requires development of alternative sources such as heterologous biosynthesis by recombinant bacteria. For this purpose, we molecularly engineered Escherichia coli to optimize the synthesis of zeaxanthin from lycopene using fusion protein-mediated substrate channeling as well as by the introduction of tunable intergenic regions. The tunable intergenic regions approach was more efficient compared with protein fusion for coordinating expression of lycopene ß-cyclase gene crtY and ß-carotene 3-hydroxylase gene crtZ. The influence of the substrate channeling effect suggests that the reaction catalyzed by CrtZ is the rate-limiting step in zeaxanthin biosynthesis. Then Pantoea ananatis, Pantoea agglomerans and Haematococcus pluvialis crtZ were compared. Because P. ananatis crtZ is superior to that of P. agglomerans or H. pluvialis for zeaxanthin production, we used it to generate a recombinant strain of E. coli BETA-1 containing pZSPBA-2(P37-crtZPAN) that produced higher amounts of zeaxanthin (11.95 ± 0.21 mg/g dry cell weight) than other engineered E. coli strains described in the literature.