Cobalt/Iron Bimetallic Biochar Composites for Lead(II) Adsorption: Mechanism and Remediation Performance.

The performance of nano-zero-valent iron for heavy metal remediation can be enhanced via incorporation into bimetallic carbon composites. However, few economical and green approaches are available for preparing bimetallic composite materials. In this study, novel Co/Fe bimetallic biochar composites (BC@Co/Fe-X, where X = 5 or 10 represents the CoCl2 concentration of 0.05 or 0.1 mol L-1) were prepared for the adsorption of Pb2+. The effect of the concentration of cross-linked metal ions on Pb2+ adsorption was investigated, with the composite prepared using 0.05 mol L-1 Co2+ (BC@Co/Fe-5) exhibiting the highest adsorption performance. Various factors, including the adsorption period, Pb2+ concentration, and pH, affected the adsorption of Pb2+ by BC@Co/Fe-5. Further characterisation of BC@Co/Fe-5 before and after Pb2+ adsorption using methods such as X-ray diffraction and X-ray photoelectron spectroscopy suggested that the Pb2+ adsorption mechanism involved (i) Pb2+ reduction to Pb0 by Co/Fe, (ii) Co/Fe corrosion to generate Fe2+ and fix Pb2+ in the form of PbO, and (iii) Pb2+ adsorption by Co/Fe biochar. Notably, BC@Co/Fe-5 exhibited excellent remediation performance in simulated Pb2+-contaminated water and soil with good recyclability.

[Effects of branched-chain amino acid supplementation on skeletal muscle proteolytic pathways after exercise-induced muscle injury in rats].

OBJECTIVE: To explore the role of branched-chain amino acid(BCAA) supplementation on muscle damage and the regulation of Krüppel-like factor 15(KLF15) and nuclear factor kappa B(NF-κB) mediated proteolytic pathways after an acute eccentric exercise. METHODS: Male SD rats were divided into placebo group(PLA) and BCAA group(BCAA) randomly, 32 rats per group. Both group were then placed into subgroups: placebo and pre-exercise group(PC), placebo and immediately after exercise group(PE), placebo and 6 h after exercise group(PE6), placebo and 12 h after exercise group(PE12), BCAA and pre-exercise group(BC), BCAA and immediately after exercise group(BE), BCAA and 6 h after exercise group(BE6), BCAA and 12 h after exercise group(BE12), 8 rats per group. Rats in BCAA groups were supplied with BCAA(1 g/(kg·d·BW), 3 days) before the exercise day and placebo groups with equal volume of distilled water. The exercised groups performed a 2 h eccentric exercise on treadmill(16 m/min, -16° slope). Blood and gastrocnemius were collected according to the time points. RT-qPCR was used to measure the mRNA expression of KLF15, NF-κB, FoxO1, Atrogin-1 and MuRF1 in gastrocnemius. RESULTS: (1) No damage was found in myocytes of BC and PC group. The process of morphological damage in BCAA group was relatively faster. (2) The mRNA expression levels of KLF15, FoXO1, Atrogin-1 and MuRF1 in PE were higher than those in PC(P<0.05, P<0.01), NF-κB and Atrogin-1 in PE12 were higher than those in PC(P<0.05). The mRNA expression levels of FoXO1 in BE were higher than those in BC(P<0.05). Compared with PE, the mRNA expression levels of KLF15, Atrogin-1 and MuRF1 in BE were lower(P<0.05, P<0.01), NF-κB and Atrogin-1 in BE12 were lower than those in PE12(P<0.05). The level of serum 3-MH in PE12 group was higher than that in PC group(P<0.05). CONCLUSION: The proteolysis of skeletal muscle after high-intensity eccentric exercise is mediated by two different pathways: KLF15 and NF-κB, whose activation is time-dependent. BCAA may reduce skeletal muscle proteolysis by lowering the level of gene transcription in the KLF15 and NF-κB related protein degradation pathway, which occurs immediately after exercise.

BL02, a phage against carbapenem- and polymyxin-B resistant Klebsiella pneumoniae, isolated from sewage: A preclinical study.

The emergence of Carbapenem-resistant Klebsiella pneumoniae (CRKP) represents a threat to public health. Polymyxin-B is generally considered a last-resort antibiotic. In this study, we isolated a carbapenem- and polymyxin-B resistant K. pneumoniae phage BL02 for the first time in Southwestern China and evaluated its biological characteristics and whole-genome sequence. Polymyxin-B resistant K. pneumoniae, (CK02), was isolated from the blood of a male with severe septic shock, and phage BL02 was screened and purified from the hospital sewage. BL02 could lyse 40 out of 46 CRKP isolates (86.96%) and has high activity in the pH range of 6-10 and the temperature range of 4-55 °C. The latency period of BL02 was about 10 min and the lysis period was about 50 min. The genome results showed that BL02 was a linear dsDNA with a total length of 175,595 bp and a GC content of 41.83%. A total of 275 ORFs were predicted and no tRNA, rRNA, drug resistance genes, or virulence genes were found in the genome. Phylogenetic analysis showed that BL02 belongs to the family Straboviridae. Treatment of infected mice with two antibiotics (tigecycline or ceftazidime/avibactam) resulted in 7-day survival rates of 28.57% and 42.86%, respectively. In contrast, the survival rate of mice in the single-dose BL02-treated group was 71.43%. In summary, this preclinical study isolated a phage capable of lysing polymyxin-B resistant K. pneumoniae and validated its safety and efficacy in an in vivo model, which provides a reference for further research on controlling MDR pathogens.

An increase in surface hydrophobicity mediates chaperone activity in N-chlorinated RidA.

Under physiological conditions, Escherichia coli RidA is an enamine/imine deaminase, which promotes the release of ammonia from reactive enamine/imine intermediates. However, when modified by hypochlorous acid (HOCl), it turns into a potent chaperone-like holdase that can effectively protect E. coli's proteome during oxidative stress. However, it is unknown, which residues need to be chlorinated for activation. Here, we employ a combination of LC-MS/MS analysis, a chemo-proteomic approach, and a mutagenesis study to identify residues responsible for RidA's chaperone-like function. Through LC-MS/MS of digested RidAHOCl, we obtained direct evidence of the chlorination of one arginine residue. To overcome the instability of the N-chloramine modification, we established a chemoproteomic approach using 5-(dimethylamino) naphthalene-1-sulfinic acid (DANSO2H) as a probe to label N-chlorinated lysines. Using this probe, we were able to detect the N-chlorination of six additional lysine residues. Moreover, using a mutagenesis study to genetically probe the role of single arginine and lysine residues, we found that the removal of arginines R105 and/or R128 led to a substantial reduction of RidAHOCl's chaperone activity. These results, together with structural analysis, confirm that the chaperone activity of RidA is concomitant with the loss of positive charges on the protein surface, leading to an increased overall protein hydrophobicity. Molecular modelling of RidAHOCl and the rational design of a RidA variant that shows chaperone activity even in the absence of HOCl further supports our hypothesis. Our data provide a molecular mechanism for HOCl-mediated chaperone activity found in RidA and a growing number of other HOCl-activated chaperones.

Thiol-based chemical probes exhibit antiviral activity against SARS-CoV-2 via allosteric disulfide disruption in the spike glycoprotein.

The development of small-molecules targeting different components of SARS-CoV-2 is a key strategy to complement antibody-based treatments and vaccination campaigns in managing the COVID-19 pandemic. Here, we show that two thiol-based chemical probes that act as reducing agents, P2119 and P2165, inhibit infection by human coronaviruses, including SARS-CoV-2, and decrease the binding of spike glycoprotein to its receptor, the angiotensin-converting enzyme 2 (ACE2). Proteomics and reactive cysteine profiling link the antiviral activity to the reduction of key disulfides, specifically by disruption of the Cys379-Cys432 and Cys391-Cys525 pairs distal to the receptor binding motif in the receptor binding domain (RBD) of the spike glycoprotein. Computational analyses provide insight into conformation changes that occur when these disulfides break or form, consistent with an allosteric role, and indicate that P2119/P2165 target a conserved hydrophobic binding pocket in the RBD with the benzyl thiol-reducing moiety pointed directly toward Cys432. These collective findings establish the vulnerability of human coronaviruses to thiol-based chemical probes and lay the groundwork for developing compounds of this class, as a strategy to inhibit the SARS-CoV-2 infection by shifting the spike glycoprotein redox scaffold.

Exploring the NT-proBNP expression in Premature Infants with Patent Ductus Arteriosus (PDA) by Echocardiography.

OBJECTIVES: To investigate the correlation between echocardiographic indicators and the expression level of N-terminal pro-brain natriuretic peptide (NT-proBNP) in premature infants (PIs) with patent ductus arteriosus (PDA) and the value of NT-proBNP in diagnosing symptomatic PDA (sPDA) in PIs whose gestational age (GA) was less than 32 weeks. METHODS: Ninty premature infants were selected as the research objects, including 52 in the non-PDA group and 38 in the PDA group (26 sPDA cases and 12 cases with asymptomatic PDA (asPDA)) from February 2019 to March 2020. The general information of these infants was recorded, including gender, delivery method, maternal infection, and serum NT-proBNP level on the 3rd day after birth. They were screened by echocardiographic indicators under an artificial intelligence convolutional neural network (AI-CNN). The Receiver Operating Characteristic (ROC) curves were illustrated to decide serum NT-proBNP expression levels, thereby determining specificity and sensitivity of sPDA and the correlation between serum sPDA NT-proBNP expression and echocardiographic indicators. RESULTS: The expression level of serum NT-proBNP in the sPDA group was greater than that in the asPDA group and the non-PDA group (P<0.001). The serum NT-proBNP expression level was positively correlated with the diameter of the ductus arteriosus in the sPDA group (r=0.462, P<0.001); it was also positively correlated with the ratio of left atrium/aorta (LA/AO) (r=0.573, P<0.001), but was not correlated with left ventricular ejection fraction (LVEF) (r=-0.015, P=0.747). CONCLUSION: The combination of serum NT-proBNP expression and echocardiography had clinical values in early diagnosis of PDA.

Application of block matching method-based Echocardiography combined with serum NT-PROBNP level detection in the early prediction of PDA in premature infants.

OBJECTIVES: The paper uses block matching method combined with echocardiography to explore the value of N-terminal pro-brain natriuretic peptide (NT-proBNP) in predicting symptomatic patent ductus arteriosus (PDA) in preterm infants. METHODS: We selected premature infants born between February 2019 and March 2020, gestational age ≤32 weeks, and echocardiography within 48 hours to determine the presence of arterial ducts as the research object, monitor their clinical manifestations, and detect serum at three and five days after birth The level of NT-proBNP was checked with echocardiography, and the children were divided into PDA group and asymptomatic patent ductus arteriosus (aPDA) group according to the clinical manifestations and echocardiographic. RESULTS: The area under the ROC curve of PDA predicted by serum NT-proBNP level at 3 days after birth was 0.949, the cut-off value was 27035pg/mL, the sensitivity was 92.3%, and the specificity was 94.6%; serum NT-proBNP level at 5 days after birth predicted the ROC curve of PDA The lower area is 0.924, the critical value is 6411 pg/mL, the sensitivity is 92.3%, and the specificity is 92.9%. CONCLUSION: NT-proBNP may be a quantitative indicator of arterial duct shunt; the detection of serum NT-proBNP levels at 3 and 5 days after birth is helpful for early prediction of PDA.

Dual-focusing behavior of a one-dimensional quadratically chirped Pearcey-Gaussian beam.

In this paper, we provide analytical solutions describing the dynamic behavior of the Pearcey-Gaussian beams propagating in free space. Based on the analytical solutions, explicit expressions governing the focusing distances of the Pearcey-Gaussian beams are found and verified by numerical simulations. For the linearly chirped Pearcey-Gaussian beam, it exhibits a uni-focusing behavior during propagation. Particularly, the focusing distance is independent on the linear chirp parameter and remains zf = 2 unchanged. Of particular interest is that the quadratically chirped Pearcey-Gaussian beam focuses twice when the quadratic chirp parameter ß < 0. The first and the second focusing distances are determined by zf1 = 2/(1 - 4ß) and zf2 = -1/(2ß), respectively. Furthermore, we numerically investigate the peak powers at the different focusing positions and find that as ß increases, the peak powers at zf1 and zf2 linearly decrease. It is expected that the characteristics can be used for manipulating the focusing distances and the peak powers to generate an optical beam with high peak power by adjusting the chirp parameter ß.

Wittig reagents for chemoselective sulfenic acid ligation enables global site stoichiometry analysis and redox-controlled mitochondrial targeting.

Triphenylphosphonium ylides, known as Wittig reagents, are one of the most commonly used tools in synthetic chemistry. Despite their considerable versatility, Wittig reagents have not yet been explored for their utility in biological applications. Here we introduce a chemoselective ligation reaction that harnesses the reactivity of Wittig reagents and the unique chemical properties of sulfenic acid, a pivotal post-translational cysteine modification in redox biology. The reaction, which generates a covalent bond between the ylide nucleophilic α-carbon and electrophilic γ-sulfur, is highly selective, rapid and affords robust labelling under a range of biocompatible reaction conditions, which includes in living cells. We highlight the broad utility of this conjugation method to enable site-specific proteome-wide stoichiometry analysis of S-sulfenylation and to visualize redox-dependent changes in mitochondrial cysteine oxidation and redox-triggered triphenylphosphonium generation for the controlled delivery of small molecules to mitochondria.

Different Infection Profiles and Antimicrobial Resistance Patterns Between Burn ICU and Common Wards.

Infection is the leading cause of complications and deaths after burns. However, the difference in infection patterns between the burn intensive care unit (BICU) and burn common wards (BCW) have not been clearly investigated. The present study aimed to compare the infection profile, antimicrobial resistance, and their changing patterns in burn patients in BICU and BCW. Clinical samples were analyzed between January 1, 2011, and December 31, 2019, in the Institute of Burn Research in Southwest China. The patient information, pathogen distribution, sources, and antimicrobial resistance were retrospectively collected. A total of 3457 and 4219 strains were detected in BICU and BCW, respectively. Wound secretions accounted for 86.6% and 44.9% in BCW and BICU, respectively. Compared with samples in BCW, samples in BICU had more fungi (11.8% vs. 8.1%), more Gram-negative bacteria (60.0% vs. 50.8%), and less Gram-positive bacteria (28.2% vs. 41.1%). Acinetobacter baumannii were the most common pathogen in BICU, compared with Staphylococcus aureus in BCW. S. aureus was the most frequent pathogen in wound secretions and tissues from both BICU and BCW. However, A. baumannii were the first in blood, sputum, and catheter samples from BICU. Overall, the multidrug-resistance (MDR) rate was higher in BICU than in BCW. However, the gap between BICU and BCW gradually shortened from 2011 to 2019. The prevalence of MDR A. baumannii and Klebsiella pneumonia significantly increased, especially in BCW. Furthermore, Carbapenem resistance among K. pneumoniae significantly increased in BICU (4.5% in 2011 vs. 40% in 2019) and BCW (0 in 2011 vs. 40% in 2019). However, the percentage of MDR P. aeruginosa sharply dropped from 85.7% to 24.5% in BICU. The incidence of MRSA was significantly higher in BICU than in BCW (94.2% vs. 71.0%) and stayed at a high level in BICU (89.5% to 96.3%). C. tropicalis and C. albicans were the two most frequent fungi. No resistance to Amphotericin B was detected. Our study shows that the infection profile is different between BICU and BCW, and multidrug resistance is more serious in BICU than BCW. Therefore, different infection-control strategies should be emphasized in different burn populations.

Parallel evaluation of nucleophilic and electrophilic chemical probes for sulfenic acid: Reactivity, selectivity and biocompatibility.

Cysteine sulfenic acids (Cys-SOH) are pivotal modifications in thiol-based redox signaling and central intermediates en route to disulfide and sulfinic acid states. A core mission in our lab is to develop bioorthogonal chemical tools with the potential to answer mechanistic questions involving cysteine oxidation. Our group, among others, has contributed to the development of nucleophilic chemical probes for detecting sulfenic acids in living cells. Recently, another class of Cys-SOH probes based on strained alkene and alkyne electrophiles has emerged. However, the use of different models of sulfenic acid and methodologies, has confounded clear comparison of these probes with respect to chemical reactivity, kinetics, and selectivity. Here, we perform a parallel evaluation of nucleophilic and electrophilic chemical probes for Cys-SOH. Among the key findings, we demonstrate that a probe for Cys-SOH based on the norbornene scaffold does not react with any of the validated sulfenic acid models in this study. Furthermore, we show that purported cross-reactivity of dimedone-like probes with electrophiles, like aldehydes and cyclic sulfenamides, is a not meaningful in a biological setting. In summary, nucleophilic probes remain the most viable tools for bioorthogonal detection of Cys-SOH.

Safety and Efficacy of a Phage, kpssk3, in an in vivo Model of Carbapenem-Resistant Hypermucoviscous Klebsiella pneumoniae Bacteremia.

Antimicrobial resistance (AMR) is one of the most significant threats to global public health. As antibiotic failure is increasing, phages are gradually becoming important agents in the post-antibiotic era. In this study, the therapeutic effects and safety of kpssk3, a previously isolated phage infecting carbapenem-resistant hypermucoviscous Klebsiella pneumoniae (CR-HMKP), were evaluated in a mouse model of systemic CR-HMKP infection. The therapeutic efficacy experiment showed that intraperitoneal injection with a single dose of phage kpssk3 (1 × 107 PFU/mouse) 3 h post infection protected 100% of BALB/c mice against bacteremia induced by intraperitoneal challenge with a 2 × LD100 dose of NY03, a CR-HMKP clinical isolate. In addition, mice were treated with antibiotics from three classes (polymyxin B, tigecycline, and ceftazidime/avibactam plus aztreonam), and the 7 days survival rates of the treated mice were 20, 20, and 90%, respectively. The safety test consisted of 2 parts: determining the cytotoxicity of kpssk3 and evaluating the short- and long-term impacts of phage therapy on the mouse gut microbiota. Phage kpssk3 was shown to not be cytotoxic to mammalian cells in vitro or in vivo. Fecal samples were collected from the phage-treated mice at 3 time points before (0 day) and after (3 and 10 days) phage therapy to study the change in the gut microbiome via high-throughput 16S rDNA sequence analysis, which revealed no notable alterations in the gut microbiota except for decreases in the Chao1 and ACE indexes.

One-pot synthesis of spherical nanoscale zero-valent iron/biochar composites for efficient removal of Pb(ii).

In this study, a spherical Fe/C composite (AIBC) was successfully prepared via carbonization of Fe3+-crosslinked sodium alginate. The removal capacity and mechanism of AIBC were evaluated for the adsorption of Pb(ii) from aqueous solution and compared with that of commercial nanoscale zero-valent iron (nZVI). The effects of the initial concentration, pH of Pb(ii) solution, the contact time, coexisting anions, and aging under air were investigated. The results showed that the pH had a strong impact on the adsorption of Pb(ii) by AIBC. The adsorption data for AIBC followed the Langmuir model, while the maximum adsorption capacity at pH 5 was 1881.73 mg g-1. The AIBC had a higher adsorption capability than nZVI, especially under the condition of relatively high Pb(ii) concentrations. The oxidation-reduction reaction between Fe and Pb(ii) was the main mechanism for the adsorption of Pb(ii) onto nZVI. AIBC converted the largest amount of Pb(ii) into PbO·XH2O/Pb(OH)2 mainly by generating Fe2+.

Activity-Based Sensing for Site-Specific Proteomic Analysis of Cysteine Oxidation.

Oxidative post-translational modifications (OxiPTMs) of cysteine residues are the molecular foundation of thiol-based redox regulation that modulates physiological events such as cell proliferation, differentiation, and migration and, when dysregulated, can lead to biomolecule damage and cell death. Common OxiPTMs of cysteine thiols (-SH) include reversible modifications such as S-sulfenylation (-SOH), S-glutathionylation (-SSG), disulfide formation (-SSR), S-nitrosylation (-SNO), and S-sulfhydration (-SSH) as well as more biologically stable modifications like S-sulfinylation (-SO2H) and S-sulfonylation (-SO3H). In the past decade, our laboratory has developed first-in-class chemistry-based tools and proteomic methods to advance the field of thiol-based redox biology and oxidative stress. In this Account, we take the reader through the historical aspects of probe development and application in our laboratory, highlighting key advances in our understanding of sulfur chemistry, in the test tube and in living systems. Offering superior resolution, throughput, accuracy, and reproducibility, mass spectrometry (MS)-based proteomics coupled to chemoselective "activity-based" small-molecule probes is the most rigorous technique for global mapping of cysteine OxiPTMs. Herein, we describe the evolution of this field from indirect detection to state-of-the-art site-centric quantitative chemoproteomic approaches that enable mapping of physiological and pathological changes in cysteine oxidation. These methods enable protein and site-level identification, mechanistic studies, mapping fold-changes, and modification stoichiometry. In particular, this Account focuses on activity-based methods for profiling S-sulfenylation, S-sulfinylation, and S-sulfhydration with an eye toward new reactions and methodologies developed in our group as well as their applications that have shed new light on fundamental processes of redox biology. Among several classes of sulfenic acid probes, dimedone-based C-nucleophiles possess superior chemical selectivity and compatibility with tandem MS. Cell-permeable dimedone derivatives with a bioconjugation handle are capable of detecting of S-sulfenylation in living cells. In-depth screening of a C-nucleophile library has yielded several entities with significantly enhanced reactivity over dimedone while maintaining selectivity, and reversible linear C-nucleophiles that enable controlled target release. C-Nucleophiles have also been implemented in tag-switch methods to detect S-sulfhydration. Most recently, activity-based detection of protein S-sulfinylation with electrophilic nitrogen species (ENS), such as C-nitroso compounds and electron deficient diazines, offers significant advantages in simplicity-of-use and target specificity compared to label-free methods. When feasible, the rich information provided by site-centric quantitative proteomics should not be tainted by oxidation artifacts from cell lysis. Therefore, chemoselective probes that function in a native environment with low cytotoxicity, good cell-permeability, and competitive kinetics are desired in modern redox chemoproteomics approaches. As our understanding of sulfur chemistry and redox signaling evolves, newly discovered cysteine OxiPTMs in microorganisms, plants, cells, tissues, and disease models should innovatively promote mechanistic and therapeutic research.

Two novel aromatic hydrocarbons: facile synthesis, photophysical properties and applications in deep-blue electroluminescence.

Two efficient novel fluorescent naphthalene and fluorene-based aromatic hydrocarbon isomers (1 and 2) are prepared and investigated for organic electroluminescence. These compounds show bright violet to deep-blue emission, narrow full width at half maximum (52 nm), and high photoluminescence efficiency (e.g. 0.61 in CH2Cl2, 0.67 in film). Alternation of substituent position on the naphthalene moiety can give rise to remarkable emission variation. The relatively large torsion angle between naphthalene and fluorene suppresses the π-π interactions by weakening the intermolecular interactions in the solid state, which can result in highly efficient fluorescence. Moreover, the 1931 Commission Internationale de L'Eclairage coordinates and maximum emission peak for deep-blue electroluminescence based on 1 are (0.16, 0.08) and 410 nm, respectively.

Characterization and genome sequencing of a novel T7-like lytic phage, kpssk3, infecting carbapenem-resistant Klebsiella pneumoniae.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) has spread globally and emerged as an urgent public health threat. Bacteriophages are considered an effective weapon against multidrug-resistant pathogens. In this study, we report a novel lytic phage, kpssk3, which is able to lyse CRKP and degrade exopolysaccharide (EPS). The morphological characteristics of kpssk3 observed by transmission electron microscopy, including a polyhedral head and a short tail, indicate that it belongs to the family Podoviridae. A one-step growth curve revealed that kpssk3 has a latent period of 10 min and a burst size of 200 plaque-forming units (pfu) per cell. kpssk3 was able to lyse 25 out of 27 (92.59%) clinically isolated CRKP strains, and it also exhibited high stability to changes in temperature and pH. kpssk3 has a linear dsDNA genome of 40,539 bp with 52.80% G+C content and 42 putative open reading frames (ORFs). No antibiotic resistance genes, virulence factors, or integrases were identified in the genome. Based on bioinformatic analysis, the tail fiber protein of phage kpssk3 was speculated to possess depolymerase activity towards EPS. By comparative genomics and phylogenetic analysis, it was determined that kpssk3 is a new T7-like virus and belongs to the subfamily Autographivirinae. The characterization and genomic analysis of kpssk3 will promote our understanding of phage biology and diversity and provide a potential strategy for controlling CRKP infection.

Lytic Bacteriophage Screening Strategies for Multidrug-Resistant Bloodstream Infections in a Burn Intensive Care Unit.

BACKGROUND Increasing antibiotic resistance and multidrug resistance (MDR) in patients with bloodstream infection (BSI) has resulted in treatment using bacteriophage. This study aimed to identify Gram-negative bacilli and Gram-positive cocci and antibiotic resistance in patients with BSI in a burn intensive care unit (BICU). The environment, including sewage systems, were investigated for the presence of lytic bacteriophage. MATERIAL AND METHODS Between January 2011 to December 2017, 486 patients with BSI were admitted to the BICU. Blood culture identified the main infectious organisms. Bacterial screening tests for antibiotic resistance included the D test and the modified Hodge test (MHT). Lytic bacteriophage was isolated from the environment. RESULTS In 486 patients with BSI, the main causative organisms were Gram-negative bacilli (64.6%), Gram-positive cocci (27.7%), and fungi (7.7%). The main pathogenic organisms that showed multidrug resistance (MDR) were Acinetobacter baumannii (26.0%), Staphylococcus aureus (16.8%), and Pseudomonas aeruginosa (14.2%). Bacteriophage was mainly isolated from Gram-negative bacilli. Screening of hospital and residential sewage systems identified increased levels of bacteriophage in hospital sewage. CONCLUSIONS The causative organisms of BSI and the presence of MDR in a hospital BICU were not typical, which supports the need for routine bacterial monitoring. Hospital sewage provides a potential source of bacteriophage for the treatment of MDR pathogenic bacteria.

Effect of Freezing on Photosystem II and Assessment of Freezing Tolerance of Tea Cultivar.

Freezing tolerant tea cultivars are urgently needed. The tea cultivars with highly freezing tolerance showed resistance to freezing stress induced photoinhibition. Freezing sensitivity index (H) of 47 tea clonal cultivars was investigated after severe freezing winter in 2016. To develop instrumental methods for freezing tolerance selection, the maximum photochemical efficiency of photosystem II (PSII) (Fv/Fm) and leaf color indicator a on the Hunter color scale were determined on control group (non-frozen) and frozen group (being frozen at -15 °C for 2 h and then stood at 20 °C for 5 h) of the cultivars. When the two indicators were expressed as the ratios (RFv/Fm and Ra) of frozen group to control group, linear regression of the freezing sensitivity index (H) upon the RFv/Fm and Ra produced significant relationship respectively, i.e., H = 60.31 - 50.09 RFv/Fm (p < 0.01) and H = 30.03 - 10.82 Ra (p < 0.01). Expression of gene psbA encoding D1 protein and gene psbD encoding D2 protein in PSII showed that the frezzing tolerant tea cultivars maintained a high expression level of psbA after freezing stress, which is considered to be beneficial to de novo synthesis of D1 protein and sustaining PSII activity. These findings can provide instrumental tools for assessing freezing tolerance of tea cultivars in tea breeding program.

Involvement of Salicylic Acid in Anthracnose Infection in Tea Plants Revealed by Transcriptome Profiling.

Anthracnose is a major leaf disease in tea plant induced by Colletotrichum, which has led to substantial losses in yield and quality of tea. The molecular mechanism with regards to responses or resistance to anthracnose in tea remains unclear. A de novo transcriptome assembly dataset was generated from healthy and anthracnose-infected leaves on tea cultivars "Longjing-43" (LJ43) and "Zhenong-139" (ZN139), with 381.52 million pair-end reads, encompassing 47.78 billion bases. The unigenes were annotated versus Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), National Center for Biotechnology Information (NCBI) non-redundant protein sequences (Nr), evolutionary genealogy of genes: Non-supervised Orthologous Groups (eggNOG) and Swiss-prot. The number of differential expression genes (DEGs) detected between healthy and infected leaves was 1621 in LJ43 and 3089 in ZN139. The GO and KEGG enrichment analysis revealed that the DEGs were highly enriched in catalytic activity, oxidation-reduction, cell-wall reinforcement, plant hormone signal transduction and plant-pathogen interaction. Further studies by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and high-performance liquid chromatography (HPLC) showed that expression of genes involved in endogenous salicylic acid biosynthesis and also accumulation of foliar salicylic acid are involved in the response of tea plant to anthracnose infection. This study firstly provided novel insight in salicylic acid acting as a key compound in the responses of tea plant to anthracnose disease. The transcriptome dataset in this study will facilitate to profile gene expression and metabolic networks associated with tea plant immunity against anthracnose.