Promotion of apoplastic oxidative burst by artificially selected GhCBSX3A enhances Verticillium dahliae resistance in upland cotton.

Verticillium wilt (VW) is a devasting disease affecting various plants, including upland cotton, a crucial fiber crop. Despite its impact, the genetic basis underlying cotton's susceptibility or defense against VW remains unclear. Here, we conducted a genome-wide association study on VW phenotyping in upland cotton and identified a locus on A13 that is significantly associated with VW resistance. We then identified a cystathionine ß-synthase domain gene at A13 locus, GhCBSX3A, which was induced by Verticillium dahliae. Functional analysis, including expression silencing in cotton and overexpression in Arabidopsis thaliana, confirmed that GhCBSX3A is a causal gene at the A13 locus, enhancing SAR-RBOHs-mediated apoplastic oxidative burst. We found allelic variation on the TATA-box of GhCBSX3A promoter attenuated its expression in upland cotton, thereby weakening VW resistance. Interestingly, we discovered that altered artificial selection of GhCBSX3A_R (an elite allele for VW) under different VW pressures during domestication and other improved processes allows specific human needs to be met. Our findings underscore the importance of GhCBSX3A in response to VW, and we propose a model for defense-associated genes being selected depending on the pathogen's pressure. The identified locus and gene serve as promising targets for VW resistance enhancement in cotton through genetic engineering.

Causal impact of DNA methylation on refracture in elderly individuals with osteoporosis - a prospective cohort study.

BACKGROUND: Osteoporotic vertebral compression fractures (OVCF) in the elderly increase refracture risk post-surgery, leading to higher mortality rates. Genome-wide association studies (GWAS) have identified susceptibility genes for osteoporosis, but the phenotypic variance explained by these genes has been limited, indicating the need to explore additional causal factors. Epigenetic modifications, such as DNA methylation, may influence osteoporosis and refracture risk. However, prospective cohorts for assessing epigenetic alterations in Chinese elderly patients are lacking. Here, we propose to conduct a prospective cohort study to investigate the causal network of DNA polymorphisms, DNA methylation, and environmental factors on the development of osteoporosis and the risk of refracture. METHODS: We will collect vertebral and peripheral blood from 500 elderly OVCF patients undergoing surgery, extract DNA, and generate whole genome genotype data and DNA methylation data. Observation indicators will be collected and combined with one-year follow-up data. A healthy control group will be selected from a natural population cohort. Epigenome-wide association studies (EWAS) of osteoporosis and bone mineral density will be conducted. Differential methylation analysis will compare candidate gene methylation patterns in patients with and without refracture. Multi-omics prediction models using genetic variants and DNA methylation sites will be built to predict OVCF risk. DISCUSSION: This study will be the first large-scale population-based study of osteoporosis and bone mineral density phenotypes based on genome-wide data, multi-time point methylation data, and phenotype data. By analyzing methylation changes related to osteoporosis and bone mineral density in OVCF patients, the study will explore the feasibility of DNA methylation in evaluating postoperative osteoporosis intervention effects. The findings may identify new molecular markers for effective anti-osteoporosis treatment and inform individualized prevention and treatment strategies. TRIAL REGISTRATION: chictr.org.cn ChiCTR2200065316, 02/11/2022.

Quantitative risk analysis of sediment heavy metals using the positive matrix factorization-based ecological risk index method: a case of the Kuye River, China.

Identifying the sources of heavy metals (HMs) in river sediments is crucial to effectively mitigate sediment HM pollution and control its associated ecological risks in coal-mining areas. In this study, ecological risks resulting from different pollution sources were evaluated using an integrated method combining the positive matrix factorization (PMF) and the potential ecological risk index (RI) model. A total of 59 sediment samples were collected from the Kuye River and analyzed for eight HMs (Zn, Cr, Ni, Cu, Pb, As, Cd, and Hg). The obtained results showed that the sediment HM contents were higher than the corresponding soil background values in Shaanxi Province. The average sediment Hg content was 3.42 times higher than the corresponding background value. The PMF results indicated that HMs in the sediments were mainly derived from industrial, traffic, agricultural, and coal-mining sources. The RI values ranged from 26.15 to 483.70. Hg was the major contributor (75%) to the ecological risk in the vicinity of the Yanjiata Industrial Park. According to the PMF-based RI model, coal-mining activities exhibited the strongest impact on the river ecosystem (48.79%), followed, respectively, by traffic (34.41%), industrial (12.70%), and agricultural (4.10%) activities. These results indicated that the major anthropogenic sources contributing to the HM contents in the sediments are not necessarily those posing the greatest ecological risks. The proposed integrated approach in this study was useful in evaluating the ecological risks associated with different anthropogenic sources in the Kuye River, providing valuable suggestions for reducing sediment HM pollution and effectively protecting river ecosystems.

Uncovering genomic and transcriptional variations facilitates utilization of wild resources in cotton disease resistance improvement.

BACKGROUND: Upland cotton wild/landraces represent a valuable resource for disease resistance alleles. Genetic differentiation between genotypes, as well as variation in Verticillium wilt (VW) resistance, has been poorly characterized for upland cotton accessions on the domestication spectrum (from wild/landraces to elite lines). RESULTS: To illustrate the effects of modern breeding on VW resistance in upland cotton, 37 wild/landraces were resequenced and phenotyped for VW resistance. Genomic patterns of differentiation were identified between wild/landraces and improved upland cotton, and a significant decline in VW resistance was observed in association with improvement. Four genotypes representing different degrees of improvement were used in a full-length transcriptome analysis to study the genetic basis of VW resistance. ROS signaling was highly conserved at the transcriptional level, likely providing the basis for VW resistance in upland cotton. ASN biosynthesis and HSP90-mediated resistance moderated the response to VW in wild/landraces, and loss of induction activity of these genes resulted in VW susceptibility. The observed genomic differentiation contributed to the loss of induction of some important VW resistance genes such as HSP90.4 and PR16. CONCLUSIONS: Besides providing new insights into the evolution of upland cotton VW resistance, this study also identifies important resistance pathways and genes for both fundamental research and cotton breeding.

[Analysis of COL1A1 and COL1A2 gene variants in two fetuses with osteogenesis imperfecta].

OBJECTIVE: To explore the genetic basis of two fetuses with an osteogenesis imperfecta (OI) phenotype. METHODS: Two fetuses diagnosed at the Affiliated Hospital of Weifang Medical College respectively on June 11, 2021 and October 16, 2021 were selected as the study subjects. Clinical data of the fetuses were collected. Amniotic fluid samples of the fetuses and peripheral blood samples of their pedigree members were collected for the extraction of genomic DNA. Whole exome sequencing (WES) and Sanger sequencing were carried out to identify the candidate variants. Minigene splicing reporter analysis was used to validate the variant which may affect the pre-mRNA splicing. RESULTS: For fetus 1, ultrasonography at 17+6 weeks of gestation had revealed shortening of bilateral humerus and femurs by more than two weeks, in addition with multiple fractures and angular deformities of long bones. WES revealed that fetus 1 had harbored a heterozygous c.3949_3950insGGCATGT (p.N1317Rfs*114) variant in exon 49 of the COL1A1 gene (NM_000088.4). Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), it was classified as a pathogenic variant (PVS1+PS2+PM2_Supporting) for disrupting the downstream open reading frame resulting in premature translational termination, being de novo in origin, and lacking records in the population and disease databases.For fetus 2, ultrasonography at 23 weeks of gestation also revealed shortening of bilateral humerus and femurs by one and four weeks, respectively, in addition with bending of bilateral femurs, tibias and fibulas. Fetus 2 had harbored a heterozygous c.1557+3A>G variant in intron 26 of the COL1A2 gene (NM_000089.4). Minigene experiment showed that it has induced skipping of exon 26 from the COL1A2 mRNA transcript, resulting in an in-frame deletion (c.1504_1557del) of the COL1A2 mRNA transcript. The variant was inherited from its father and had been previously reported in a family with OI type 4. It was therefore classified as a pathogenic variant (PS3+PM1+PM2_Supporting+PP3+PP5). CONCLUSION: The c.3949_3950insGGCATGT (p.N1317Rfs*114) variant in the COL1A1 gene and c.1557+3A>G variant in the COL1A2 gene probably underlay the disease in the two fetuses. Above findings not only have enriched the mutational spectrum of OI, but also shed light on the correlation between its genotype and phenotype and provided a basis for genetic counseling and prenatal diagnosis for the affected pedigrees.

A meta-analysis comparing the effects of cemented and uncemented prostheses on wound infection and pain in patients with femoral neck fractures.

To providing evidence-based recommendations for surgery in patients with femoral neck fractures, a meta-analysis was conducted to comprehensively evaluate the effects of cemented and uncemented prostheses on postoperative surgical site wound infection and pain in these patients. Relevant studies on the use of cemented prostheses in femoral neck fractures were retrieved from PubMed, EMBASE, Cochrane Library, Ovid, CNKI, and Wanfang databases from the time of their establishment until March 2023. Two authors independently screened and extracted data from the included and excluded literature according to predetermined criteria. Review Manager 5.4 software was used to perform meta-analyses on the collected data. A total of 27 articles comprising 34 210 patients (24 646 cases in the cemented group and 9564 cases in the uncemented group) were included in the final analysis. The results of the meta-analysis showed that, compared with the uncemented group, cemented prostheses significantly reduced the incidence of surgical site wound infections (odds ratio [OR]: 0.75, 95% confidence interval [CI]: 0.64-0.88, p < 0.001) and relieved surgical site wound pain (standardised mean difference: -0.76, 95% CI: -1.12-0.40, p < 0.001), but did not reduce the incidence of pressure ulcers after surgery (OR: 0.50, 95% CI: 0.20-1.26, p = 0.140). Therefore, existing evidence suggests that the use of cemented prostheses in femoral neck fracture surgery can significantly reduce the incidence of surgical site wound infections and relieve surgical site wound pain, which is worthy of clinical recommendation.

Integrating Interactive Noble Metal Single-Atom Catalysts into Transition Metal Oxide Lattices.

Noble metals have broad prospects for catalytic applications yet are restricted to a few packing modes with limited structural flexibility. Here, we achieved geometric structure diversification of noble metals by integrating spatially correlated noble metal single atoms (e.g., Pt, Pd, and Ru) into the lattice of transition metal oxides (TMOs, e.g., Co3O4, Mn5O8, NiO, Fe2O3). The obtained noble metal single atoms exhibited distinct topologies (e.g., crs, fcu-hex-pcu, fcu, and bcu-x) from those of conventional metallic phases. For example, Pt single atoms with a crs topology (Ptcrs-Co3O4) are endowed with synergy of metal-metal and metal-support interactions. A quantitative relationship between various Pt topologies determined by TMO substrates and their electrocatalytic activities was established. We anticipate that this type of interactive single-atom catalysts can bridge the geometric, topological, and electronic structure gaps between the "close-packed" nanoparticles and isolated single atoms as two common categories of heterogeneous catalysts.

Racial and ethnic disparities in nasopharyngeal cancer with an emphasis among Asian Americans.

Despite the overall decreasing incidence, nasopharyngeal cancer (NPC) continues to cause a significant health burden among Asian Americans (AAs), who are a fast-growing but understudied heterogeneous racial group in the United States. We aimed to examine the racial/ethnic disparities in NPC incidence, treatment, and mortality with a specific focus on AA subgroups. NPC patients aged ≥15 years were obtained from the Surveillance, Epidemiology, and End Results (SEER) 18 (1975-2018). AAs were divided into Chinese, Filipino, Vietnamese, Hawaiian, Japanese, Laotian, Korean, Cambodian, Indian/Pakistani and other Asian/Pacific Islanders (APIs). Age-adjusted incidence was calculated using the SEER*Stat software. Cox proportional and Fine-Gray subdistribution hazard models were used to calculate overall and cause-specific mortalities after adjusting for confounders. Among the total 11 964 NPC cases, 18.4% were Chinese, 7.7% Filipino, 5.0% Vietnamese, 1.2% Hawaiian, 1.0% Japanese, 0.8% Laotian, 0.8% Korean, 0.6% Cambodian, 0.5% Indian/Pakistani and 4.4% other APIs. Laotians had the highest age-adjusted NPC incidence (9.21 per 100 000), which was 18.04 times higher than it in non-Hispanic Whites (NHWs). Chinese and Filipinos observed lower overall mortalities, however, Chinese saw increased NPC-specific mortality than NHWs. Disparities in mortality were also found across different histology subtypes. This is the first and largest study examining the NPC incidence and outcomes in AA subgroups. The significant disparities of NPC within AAs underline the importance of adequate AA-subgroup sample size in future studies to understand the prognostic role of ethnicity in NPC and advocate more ethnically and culturally tailored cancer prevention and care delivery.

Pseudohalioglobus sediminis sp. nov., isolated from coastal sediment.

A Gram-stain-negative, strictly aerobic, non-motile and rod-shaped bacterial strain, designated NY5T, was isolated from marine sediment collected from coastal area in Weihai, China (122°07' 38.80'' E, 37°33' 57.60'' N). Cells of strain NY5T were 0.6-0.7 µm width and 1.9-2.0 µm length, catalase-positive and oxidase-positive. Growth of NY5T was observed at 25-37 °C (optimum, 28 °C) and pH 6.5-9.5 (optimum, pH 7.5-8.0) and in the presence of 0.5-7.0% (w/v) NaCl (optimum, 2.0%). The isoprenoid quinone was Q-8 and the predominant fatty acids were summed feature 8 (C18:1 ω7c and/or C18:1 ω6c), summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and C17:1 ω8c. Diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylglycerol were the major polar lipids. The DNA G+C content of strain NY5T was 60.1%. Strain NY5T showed the highest 16S rRNA gene sequence similarity (98.2%) with Pseudohalioglobus lutimaris followed by Parahaliea aestuarii (96.9%), Parahaliea maris (96.7%), Parahaliea mediterranea (95.9%), and Halioglobus japonicus (94.9%). Given these phenotypic and chemotaxonomic properties and phylogenetic analyses, strain NY5T was considered to represent a novel species of the genus Pseudohalioglobus, for which the name Pseudohalioglobus sediminis sp. nov. is proposed. The type strain is NY5T (=KCTC 72416T=MCCC 1H00401T).

Unveiling the Mechanism of Frictional Catalysis in Water by Bi12TiO20: A Charge Transfer and Contaminant Decomposition Path Study.

Tribocatalysis, as a new approach in environmental purification, has drawn increasing attention in the past few years. In this work, we successfully convert mechanical energy to chemical energy by Bi12TiO20, which was synthesized by a hydrothermal method. Under magnetic stirring, electrons transfer from the surface of Bi12TiO20 to the polytetrafluoroethylene-sealed magnetic bar due to their friction. Moreover, the holes that remain on Bi12TiO20 provide oxidation properties in the process for organic matter degradation. According to a series of tests, it is noticed that the shape of the stirring bar and the material of the reaction vessel have a remarkable influence on the removal efficiency of contaminants. Simultaneously, multiple tests reveal the high stability of Bi12TiO20. A great potential for Bi12TiO20 to control water pollutants under dark conditions during collection of ambient mechanical energy was clearly demonstrated in this study.

Adsorption-enhanced photocatalytic property of Ag-doped biochar/g-C3N4/TiO2 composite by incorporating cotton-based biochar.

In this study, the biochar obtained from waste cotton fibers was introduced into the Ag-doped g-C3N4/TiO2 hybrid composite through a facile one-step hydrothermal process. The morphology, elemental composition, crystal structure, microstructure, specific surface area, chemical bonding state, energy band structure, and separation efficiency of photoinduced charge carriers of the resultant composite were examined using scanning electron microscope, energy dispersive X-ray spectrometer, X-ray diffractometer, transmission electron microscope, surface area analyzer, X-ray photoelectron spectroscope, Ultraviolet-visible spectrophotometer, ultraviolet photoelectron spectroscope, and photoluminescence spectroscope. The adsorption isotherms, kinetics and thermodynamics of the biochar, Ag-doped g-C3N4/TiO2 and Ag-doped biochar/g-C3N4/TiO2 were evaluated using the model methyl orange dye. The photoacatalytic degradation of the model pollutants including methyl orange, methylene blue, congo red, and tetracycline hydrochloride and the photocatalytic reduction of Cr(VI) ions were also assessed under visible light. Experimental results indicated that the photocatalytic property of the Ag-doped biochar/g-C3N4/TiO2 was significantly enhanced through the adsorption enhancement compared with the Ag-doped g-C3N4/TiO2. This was due to the uniform doping of multi-scale porous biochar with g-C3N4 nanosheet, Ag and TiO2 nanoparticles. The adsorptive enhancement induced by the biochar resulted in the narrowed band gap, suitable electronic energy band structure, and fast separation of photoinduced charge carriers of the Ag-doped biochar/g-C3N4/TiO2, which was probably due to the coexistence of multi-valence Ti+4/+3 and Ag0/+1 species and oxygen-containing groups of biochar. The major reactive species of the Ag-doped biochar/g-C3N4/TiO2 were 1O2 and h+. The MO dye adsorption onto the Ag-doped biochar/g-C3N4/TiO2 followed the Langmuir isotherm model, pseudo-first-order and pseudo-second-order kinetic models, and the adsorption process was an endothermic reaction with entropy reduction effects. As such, the Ag-doped biochar/g-C3N4/TiO2 exhibited a promising application for the treatment of wastewater containing multi-pollutants especially organic dyes and heavy metal ions.

The role of IL-6, IL-10, and PGE2 in the treatment of intervertebral disc herniation by dual-channel endoscopic lumbar discectomy.

is the This study aimed to explore the role of IL-6, IL-10, and PGE2 in the treatment of intervertebral disc herniation with dual-channel endoscopic lumbar discectomy. For this purpose, we selected 182 patients with intervertebral disc herniation in our hospital and randomly divided them into the control group and the study group according to the order of admission, of which 85 cases were in the control group, 97 cases in the study group, and control group was treated with conventional lumbar discectomy; the study group was treated with dual-channel spine endoscopic lumbar discectomy to observe and compare the operation-related indexes, lumbar function indexes, clinical effects, serum-related indexes and the evaluation value of the two groups of patients. Results showed that the operation time, incision length, intraoperative blood loss, hospital stay, and postoperative pain scores of the study group were lower than those of the control group (p<0.05); the ODI and RMQ scores of the study group after treatment were lower than those of the control group (P<0.05). The excellent and good rate of the study group was 89.69% higher than that of the control group 77.65% (p<0.05); the levels of IL-6 and PGE2 in the study group after treatment were lower than those of the control group, and the IL-10 level was higher than that of the control group. (P<0.05); Using the lumbar spine function score as the comparison standard: IL-6, IL-10, PGE2 for the evaluation value of dual-channel endoscopic lumbar discectomy for the treatment of intervertebral disc herniation: sensitivity 96.18%, specificity 96.27%, the accuracy of 97.06% was higher than the single diagnosis result (P<0.05. It is worthy of clinical promotion. IL-6, IL-10, PGE2 predict dual-channel spine Endoscopic lumbar discectomy for the treatment of intervertebral disc herniation has high prognostic sensitivity and accuracy, which can provide references for clinical treatment and prognostic medication.

The effect of locating and sliding of facet combined with percutaneous endoscopic lumbar discectomy on cell inflammatory indicators and the treatment of disc herniation.

This study aimed to investigate the effects of IL-8, CRP and TXB2 in the treatment of lumbar disc herniation by combining with percutaneous endoscopic discectomy. For this purpose, 290 patients with disc herniation were selected as the research objects and randomly divided into two groups. The control group was treated with traditional intervertebral fenestration of nucleus pulposus, and the research group was treated with joint process location slip technique combined with percutaneous endoscopic lumbar disc discectomy. The clinical efficacy, functional scores and serological indexes of the two groups were compared, and the prognostic value of IL-8, CRP and TXB2 in the treatment of disc herniation by the combination of the sliding technique of facet location and percutaneous endoscopic discectomy was explored. The results showed that the total effective rate of 95.55% in the study group was higher than 79.31% in the control group, and the difference was significant (P<0.05). The operative time, incision length, length of hospital stay and intraoperative blood loss in the study group were lower than those in the control group (P<0.05). JOA score was higher and ODI score was lower in the two groups after surgery than before surgery, and JOA score in the study group was higher than that in the control group, while the ODI score was lower than that in the control group (P<0.05). Il-8, CRP and MDA in 2 groups increased after the operation, while SOD and TXB2 decreased significantly. Il-8, CRP, TXB2 and SOD in the study group were lower than those in the control group, while MDA was higher than those in the control group (P<0.05). ROC curve indicated that the areas under the curves of IL-8, CRP and TXB2 were 0.725, 0.835 and 0.880, and the areas under the curves, sensitivity and specificity of the combined determination were higher than those of any index (P<0.05). In general, compared with traditional interlaminar fenestration of nucleus pulposus, combined with percutaneous endoscopic lumbar disc discectomy has a significant effect on the treatment of disc herniation, and can reduce the levels of IL-8, CRP and TXB2.

Dietary phytoestrogen intake and lung cancer risk: an analysis of the Prostate, Lung, Colorectal and Ovarian (PLCO) cancer screening trial.

Phytoestrogens (PEs) have estrogen-like activity and were found to lower incidences of several hormone-dependent cancers. Emerging evidence suggests that estrogen may play a role in lung cancer carcinogenesis. We aim to evaluate dietary PE intake and lung cancer risk using data from the Prostate, Lung, Colorectal and Ovarian cancer screening trial. A total of 1706 lung cancer cases were identified. The association between lung cancer risk and PE intake (in quartiles) was calculated using the Cox proportional hazard models adjusting for potential confounders. Stratified analyses by smoking status, sex and histology were also performed. The highest quartile of total PE intake was associated with a reduced risk of lung cancer compared with the lowest quartile [hazard ratio (HR) = 0.85, 95% confidence interval (CI): 0.73-0.99 for >1030 µg/day versus <290 µg/day] (P trend = 0.56). Similar patterns were observed among ever smokers (HR = 0.84, 95% CI: 0.71-0.98), non-small cell histology (HR = 0.84, 95% CI: 0.72-0.99), male (HR = 0.84, 95% CI: 0.69-1.03) and female (HR = 0.80, 95% CI: 0.64-0.99 for 510-1030 µg/day, HR = 0.84, 95% CI: 0.67-1.06 for >1030 µg/day versus <290 µg/day) subjects with no significant linear trend observed. Despite a lower consumption compared with the Asian population, increased PE intake still appears to decrease lung cancer risk in a Caucasian-dominant population. Future studies are needed to replicate these results in independent cohorts and shed a light on the potential mechanism of the protective effect of PEs on lung carcinogenesis and the interaction between PEs, smoking and endogenous estrogens.

CO2 Electroreduction to Formate at a Partial Current Density up to 590 mA mg-1 via Micrometer-Scale Lateral Structuring of Bismuth Nanosheets.

2D bismuth nanosheets are a promising layered material for formate-producing via electrocatalytic CO2 conversion. However, the commercial interest of bismuth nanosheets in CO2 electroreduction is still rare due to the undesirable current density for formate at moderate operation potentials (about 200 mA mg-1 ) and harsh synthesis conditions (high temperature and/or high pressure). This work reports the preparation of Bi nanosheets with a lateral size in micrometer-scale via electrochemical cathodic exfoliation in aqueous solution at normal pressure and temperature. As-prepared Bi LNSs (L indicates large lateral size) possess high Faradaic efficiencies over 90% within a broad potential window from -0.44 to -1.10 V versus RHE and a superior partial current density about 590 mA mg-1 for formate in comparison with state-of-the-art results. Structure analysis, electrochemical results, and density functional theory calculations demonstrate that the increasing tensile lattice strain observed in Bi LNSs leads to less overlap of d orbitals and a narrower d-band width, which tuning the intermediate binding energies, and therefore promotes the intrinsic activity.

Palleronia sediminis sp. nov. and Flavivirga algicola sp. nov., two marine bacteria isolated from offshore areas near Weihai.

Two bacterial strains, designated SS33T and Y03T, were isolated from marine sediment and marine red alga collected on the coast of Weihai, PR China. Based on the results of 16S rRNA gene sequence analysis, strain SS33T was found to be closely related to Primorskyibacter marinus PX7T, Pelagivirga dicentrarchi YLY04T, Palleronia marisminoris DSM 26347T and Maribius pontilimi GH1-23T with 94.8, 94.6, 94.5 and 94.5 % sequence similarity; strain Y03T was found to be closest to Flavivirga aquimarina EC2D5T, Flavivirga eckloniae ECD14T and Flavivirga amylovorans JC2681T with 96.4, 96.1 and 96.0 % sequence similarity. Strain SS33T grew at 4-37 °C (optimum, 33 °C), at pH 6.0-9.5 (optimum, pH 7.5-8.0) and in the presence of 0-10 % (w/v) NaCl (optimum, 3.0 %). Chemotaxonomic analysis of strain SS33T showed that the predominant respiratory quinone was ubiquinone-10. The major fatty acids (>10.0 %) included C18 : 1 ω7c and C16 : 0. The major polar lipids included phosphatidylglycerol, phosphatidylcholine, one unidentified phospholipid, one unidentified glycolipid, one unidentified polar lipid and two unidentified aminolipids. Strain Y03T grew at 15-40 °C (optimum, 28 °C), at pH 6.5-8.0 (optimum, pH 7.0) and in the presence of 0.5-9.0 % (w/v) NaCl (optimum, 2.0%). Chemotaxonomic analysis showed that the predominant respiratory quinone was menaquinone-6. The major fatty acids (>10.0 %) included iso-C15 : 0, iso-C15 : 1 G, iso-C17 : 0 3-OH and iso-C15 : 0 3-OH. The major polar lipids included phosphatidylethanolamine, one unidentified phospholipid, one unidentified aminolipid and four unidentified polar lipids. Based on the polyphasic data, strain SS33T is considered to represent a novel species of the genus Palleronia, for which the name Palleronia sediminis sp. nov. is proposed, with the type strain SS33T (=KCTC 62986T=MCCC 1H00387T). Strain Y03T is considered to represent a novel species of the genus Flavivirga, for which the name Flavivirga algicola sp. nov. is proposed, with the type strain Y03T (=KCTC 72001T=MCCC 1H00386T).

Potentiometric Sensor Based on Carbon Paste Electrode for Monitoring Total Residual Chlorine in Electrolytically-Treated Ballast Water.

A new potentiometric sensor based on modified carbon paste electrode (CPE) was prepared for the sensitive and selective detection of total residual chlorine (TRC) in simulated electrolytically-treated ballast water (BW). The modified CPE was prepared using ferrocene (Fc) as the sensing species and paraffin oil as the binder. It is revealed that the addition of Fc can significantly shorten the response time and improve the reproducibility, selectivity, and stability of the sensor. The open circuit potential of the Fc-CPE is in linear proportion to the logarithm of TRC within the TRC concentration range from 1 mg∙dm-3 to 15 mg∙dm-3. In addition, the Fc-CPE sensor exhibits good selectivity to TRC over a wide concentration range of the possible co-exiting interference ions in seawater. The Fc-CPE electrode can be used as a convenient and reliable sensor for the continuous monitoring of TRC during the electrolytic treatment of BW.

Genomic and proteomic characterization of SE-I, a temperate bacteriophage infecting Erysipelothrix rhusiopathiae.

A bacteriophage infecting pathogenic Erysipelothrix rhusiopathiae was isolated from a swine farm experiencing an outbreak of acute swine erysipelas; we designated this phage SE-I. SE-I has an icosahedral head, a long tail and a double-stranded DNA genome. The 34,997-bp genome has a GC content of 34 % and contains 43 open reading frames (ORFs) encoding packaging, structural, lysin-holin, and hypothetical proteins. Components of purified SE-I were separated using SDS-PAGE and analyzed using liquid chromatography-mass spectrometry. Nine proteins were identified, encoded by ORF9, ORF15, ORF23, ORF30, ORF31, ORF33, ORF39, ORF40 and ORF 42. A phylogenetic tree constructed based on the sequence of the large terminase subunit revealed that SE-I is closely related to Staphylococcus phages P954 and phi3396. The CHAP-domain-containing protein encoded by ORF25 was expressed in E. coli and which was able to inactivate host bacteria. SE-I was able to infect 7 of 13 E. rhusiopathiae strains, but was unable to infect Salmonella, Streptococcus suis, and Staphylococcus aureus. This is the first report of the isolation, characterization, and genomic and proteomic analysis of a temperate phage infecting E. rhusiopathiae, and it might lead to the development of new anti- E. rhusiopathiae agents.

Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli.

α-Conotoxin LvIA is derived from Conus lividus, native to Hainan, and is the most selective inhibitor of α3ß2 nicotinic acetylcholine receptors (nAChRs) known to date. In this study, an efficient approach for the production of recombinant α-Conotoxin LvIA is described. Tandem repeats of a LvIA gene fragment were constructed and fused with a KSI gene and a His6 tag in a Escherichia coli (E. coli) expression vector pET-31b(+). The recombinant plasmids were transformed into E. coli and were found to express well. The KSI-(LvIA)n-His6 fusion protein was purified by metal affinity chromatography and then cleaved with CNBr to release recombinant LvIA (rLvIA). High yields of fusion protein ranging from 100 to 500 mg/L culture were obtained. The pharmacological profile of rLvIA was determined by two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing rat nAChR subtypes. The rLvIA antagonized the α3ß2 nAChR subtype selectively with a nano-molar IC50. The rLvIA was analgesic in a mouse hot-plate test model of pain. Overall, this study provides an effective method to synthesize α-conotoxin LvIA in an E. coli recombinant expression system, and this approach could be useful to obtain active conopeptides in large quantity and at low cost.

Identification of Mycoplasma suis MSG1 interaction proteins on porcine erythrocytes.

Adhesion protein MSG1 mediating adherence to porcine erythrocytes in Mycoplasma suis (M. suis) invasion has been identified previously. In order to determine the host membrane proteins that interact with MSG1, recombinant His-tagged MSG1 (rMSG1) was used to screen for interacting proteins in the protein extracts of porcine erythrocyte membrane. Potential rMSG1-interacting proteins were initially identified as band 3 and ß-actin with molecular weight of 46 and 45 kDa, respectively. Immune fluorescence results showed that rMSG1 can specifically bind with the ß-actin of HeLa, BHK-21, and HEK-293A cells, respectively. RNA interference assays further demonstrated that the interaction between ß-actin and rMSG1 on HeLa cells was specific and dose dependent. Confocal microscopy showed that both rMSG1 and M. suis can partially co-localize with ß-actin on the surface of porcine erythrocytes. Pull-down assays showed that rMSG1 can directly interact with ß-actin. Our study is the first to report the interaction of MSG1 with ß-actin, which will be of help to understand the pathogenesis of M. suis and develop a cultivation system.