Background:Treatment of methicillin-resistant Staphylococcus aureus(MRSA)biofilm infections in implant placement surgery is limited by the lack of antimicrobial activity of titanium(Ti)implants.There is a need to expl...Background:Treatment of methicillin-resistant Staphylococcus aureus(MRSA)biofilm infections in implant placement surgery is limited by the lack of antimicrobial activity of titanium(Ti)implants.There is a need to explore more effective approaches for the treatment of MRSA biofilm infections.Methods:Herein,an interfacial functionalization strategy is proposed by the integration of mesoporous polydopamine nanoparticles(PDA),nitric oxide(NO)release donor sodium nitroprusside(SNP)and osteogenic growth peptide(OGP)onto Ti implants,denoted as Ti-PDA@SNP-OGP.The physical and chemical properties of Ti-PDA@SNP-OGP were assessed by scanning electron microscopy,X-ray photoelectron spectroscope,water contact angle,photothermal property and NO release behavior.The synergistic antibacterial effect and elimination of the MRSA biofilms were evaluated by 2′,7′-dichlorofluorescein diacetate probe,1-N-phenylnaphthylamine assay,adenosine triphosphate intensity,O-nitrophenyl-β-D-galactopyranoside hydrolysis activity,bicinchoninic acid leakage.Fluorescence staining,assays for alkaline phosphatase activity,collagen secretion and extracellular matrix mineralization,quantitative real‑time reverse transcription‑polymerase chain reaction,and enzyme-linked immunosorbent assay(ELISA)were used to evaluate the inflammatory response and osteogenic ability in bone marrow stromal cells(MSCs),RAW264.7 cells and their co-culture system.Giemsa staining,ELISA,micro-CT,hematoxylin and eosin,Masson's trichrome and immunohistochemistry staining were used to evaluate the eradication of MRSA biofilms,inhibition of inflammatory response,and promotion of osseointegration of Ti-PDA@SNP-OGP in vivo.Results:Ti-PDA@SNP-OGP displayed a synergistic photothermal and NO-dependent antibacterial effect against MRSA following near-infrared light(NIR)irradiation,and effectively eliminated the formed MRSA biofilms by inducing reactive oxygen species(ROS)-mediated oxidative stress,destroying bacterial membrane integrity and causing leakage of intracellular components(P<0.01).In vitro experiments revealed that Ti-PDA@SNP-OGP not only facilitated osteogenic differentiation of MSCs,but also promoted the polarization of pro-inflammatory M1 macrophages to the anti-inflammatory M2-phenotype(P<0.05 or P<0.01).The favorable osteo-immune microenvironment further facilitated osteogenesis of MSCs and the anti-inflammation of RAW264.7 cells via multiple paracrine signaling pathways(P<0.01).In vivo evaluation confirmed the aforementioned results and revealed that Ti-PDA@SNP-OGP induced ameliorative osseointegration in an MRSA-infected femoral defect implantation model(P<0.01).Conclusions:Ti-PDA@SNP-OGP is a promising multi-functional material for the high-efficient treatment of MRSA infections in implant replacement surgeries.展开更多
Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens...Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.展开更多
Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatme...Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatment.Staphyloxanthin is a crucial virulence factor of S.aureus.Blocking staphyloxanthin production could help the host immune system counteract the invading S.aureus cells.In this study,we first screened for staphyloxanthin inhibitors using a virtual screening method.The outcome of the virtual screening method resulted in the identification of eugenol(300μg/mL),which significantly inhibits the staphyloxanthin production in S.aureus ATCC 29213,S.aureus Newman,MRSA ATCC 43300 and MRSA ATCC BAA1717by 84.2%,63.5%,68.1%,and 79.5%,respectively.The outcome of the growth curve assay,field-emission scanning electron,and confocal laser scanning microscopy analyses confirmed that eugenol at the test concentration did not affect the morphology and growth of S.aureus.Moreover,the survival rate of S.aureus ATCC 29213 and MRSA ATCC 43300 under H_(2)O_(2) pressure decreased to 51.9%and 45.5%in the presence of eugenol,respectively.The quantitative RT-PCR and molecular simulation studies revealed that eugenol targets staphyloxanthin biosynthesis by downregulating the transcription of the crtM gene and inhibiting the activity of the CrtM enzyme.Taken together,we first determined that eugenol was a prominent compound for staphyloxanthin inhibitor to combat S.aureus especially MRSA infections.展开更多
Sensitive detection of Staphylococcus aureus enterotoxin B(SEB)is of importance for preventing food poisoning from threatening human health.In this work,an electrochemical and colorimetric dual-signal detection assay ...Sensitive detection of Staphylococcus aureus enterotoxin B(SEB)is of importance for preventing food poisoning from threatening human health.In this work,an electrochemical and colorimetric dual-signal detection assay of SEB was developed.The probe(Ab2/AuPt@Fe-N-C)was bound to SEB captured by Ab1,where the Ab2/AuPt@Fe-N-C triggered methylene blue degradation and resulted in the decrease of electrochemical signal.Furthermore,the probe catalyzed the oxidation of 3,3’,5,5’-tetramethyl biphenyl to generate a colorimetric absorbance at 652 nm.Once the target was captured and formed a sandwich-like complex,the color changed from colorless to blue.SEB detection by colorimetric and electrochemical methods showed a linear relationship in the concentration ranges of 0.0002-10.0000 and 0.0005-10.0000 ng/mL,with limits of detection of 0.0667 and 0.1670 pg/mL,respectively.The dual-signal biosensor was successfully used to detect SEB in milk and water samples,which has great potential in toxin detection in food and the environment.展开更多
Objective To explore the mechanisms involved in Staphylococcus aureus （S. aureus） invading human monocytic U937 cells. Methods S. aureus were added to U937 cells at multiplicity of infections （MOI） of 20：1 for 0...Objective To explore the mechanisms involved in Staphylococcus aureus （S. aureus） invading human monocytic U937 cells. Methods S. aureus were added to U937 cells at multiplicity of infections （MOI） of 20：1 for 0, 15, 30, 60, and 90 minutes, respectively. Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI） flow cytometry analysis. Akt and nuclear factor-κB （NF-κB） activities were detected by Western blotting. Results Infection of U937 cells with S. aureus induced rapid cell death in a time-dependent manner, and the cells displayed characteristic features of apoptosis. S. aureus-induced apoptosis was associated with a prominent downregulation of activated （phosphorylated） Akt and NF-κB. The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-κB in a dose-dependent manner. Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells. Conclusions S. aureus can stimulate the apoptosis of U937 ceils. S. aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-κB.展开更多
Objective To investigate the effect of SP600125, a specific c-jun N-terminal protein kinase (JNK) inhibitor, on Staphylococcus aureus (S. aureus)-induced U937 cell death and the underlying mechanism. Methods The human...Objective To investigate the effect of SP600125, a specific c-jun N-terminal protein kinase (JNK) inhibitor, on Staphylococcus aureus (S. aureus)-induced U937 cell death and the underlying mechanism. Methods The human monocytic U937 cells were treated with S. aureus at different time with or without SP600125. Cell apoptosis was analyzed by flow cytometry. JNK, Bax, and caspase-3 activities were detected by Western blotting. Results S. aureus induced apoptosis in cultured U937 cells in a time-dependent manner. Expression of Bax and phospho-JNK significantly increased in S. aureus-treated U937 cells, and the level of activated caspase-3 also increased in a time-dependent manner. Inhibition of JNK with SP600125 significantly inhibited S. aureus-induced apoptosis in U937 cells. Conclusions S. aureus can induce apoptosis in U937 cells by phosphorylation of JNK and activation of Bax and caspase-3. SP600125 protects U937 cells from apoptosis induced by S. aureus via inhibiting the activity of JNK.展开更多
Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action mod...Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action modes of ultrasound on both Gram-negative and Gram-positive microorganisms were estimated.From colony results,ultrasound acted as an irreversible effect on both Eshcerichia coli and Staphylococcus aureus without sublethal injury.The result in this study also showed that a proportion of bacteria subpopulation suffered from serious damage of intracellular components(e.g.DNA and enzymes)but with intact cell envelopes.We speculated that the inactivated effects of ultrasound on microbes might more than simply completed disruption of cell exteriors.Those microbial cells who had not enter the valid area of ultrasonic cavitation might be injected with free radicals produced by ultrasound and experienced interior injury with intact exterior structure,and others who were in close proximity to the ultrasonic wave field would be immediately and completely disrupted into debris by high power mechanic forces.These findings here try to provide extension for the inactivation mechanisms of ultrasound on microorganisms.展开更多
This study was conducted to analyze the effects of sodium nitrite,nisin,potassium sorbate,and sodium lactate against Staphylococcus aureus(S.aureus)growth and staphylococcal enterotoxins(SEs)production in cooked pork ...This study was conducted to analyze the effects of sodium nitrite,nisin,potassium sorbate,and sodium lactate against Staphylococcus aureus(S.aureus)growth and staphylococcal enterotoxins(SEs)production in cooked pork sausage by inoculating sausage samples containing preservative with an S.aureus strain producing staphylococcal enterotoxin A(SEA)and then storing them at 37℃ for 36 h.Samples were analyzed every 3 h to count the S.aureus colonies and to detect SEA.The modified Gompertz model was used to describe S.aureus growth in the samples under various conditions,and the preservatives with a significant antimicrobial effect were selected.In addition,the antimicrobial effects of the selected preservatives under various concentrations were tested.Results showed that sodium nitrite,nisin,and potassium sorbate had a weak effect against S.aureus growth and had no effect against SEA production,whereas sodium lactate could significantly inhibit S.aureus growth and SEA production.Moreover,the antimicrobial effect of sodium lactate was concentration-dependent,wherein sodium lactate concentration<12 g/kg showed no inhibitory effect,but when the concentration was increased to 24 g/kg,sodium lactate could effectively inhibit S.aureus growth and SEA production,and at 48 g/kg,sodium lactate had a significant inhibitory effect.展开更多
In this paper,a low-pressure capacitively coupled plasma discharge sustained in an argonoxygen mixture was studied in order to evaluate its properties in terms of inactivation of Staphylococcus aureus.The plasma param...In this paper,a low-pressure capacitively coupled plasma discharge sustained in an argonoxygen mixture was studied in order to evaluate its properties in terms of inactivation of Staphylococcus aureus.The plasma parameters as electron temperature and plasma density were measured by the Langmuir probe(Ne≈1015 m^-3,Te≈1.5 eV),while the neutral atom density was in the range of 1021 m^-3.In the plasma phase,oxygen radicals were taken as reference of the reactive species with antimicrobial activity,and oxygen spectral lines,over a range of plasma process parameters,were investigated by the optical emission spectroscopy.Optimal plasma conditions were found,and a count reduction of 4 log in a few minutes of the bacterium proves the potentiality of an industrial grade plasma reactor as a sterilization agent.展开更多
BACKGROUND:Over the past two decades,methicillin-resistant Staphylococcus aureus(MRSA) has evolved from a hospital-associated infection to a significant public health threat in the community,causing outbreaks of soft ...BACKGROUND:Over the past two decades,methicillin-resistant Staphylococcus aureus(MRSA) has evolved from a hospital-associated infection to a significant public health threat in the community,causing outbreaks of soft tissue infections in otherwise healthy individuals.The goal of this study was to determine the prevalence of nasal MRSA colonization in low acuity Emergency Department(ED) Fast Track patients in order to better characterize the epidemiology of this pathogen.METHODS:We conducted a cross-sectional study of a convenience sample of adult patients from our ED Fast Track.Nasal swabs were analyzed for MRSA using a polymerase chain reaction assay.Study participants completed a survey assessing traditional risk factors for CA-MRSA colonization.RESULTS:A total of 106 ED Fast Track patients were tested.Four(3.8%,95%CI 1.5%-9.3%)were MRSA positive.Three traditional CA-MRSA risk factors(personal history of abscess,family history of abscess,and participation in contact sports) were examined.In patients with a positive MRSA nasal swab,only a personal prior history of abscess retained significance(OR 33,95%C11.7-676,P=0.02).CONCLUSION:This study found a higher prevalence of nasal MRSA colonization in low acuity ED Fast Track patients compared with historical community surveillance studies.A personal history of prior abscess was a significant risk for CA-MRSA carriage.展开更多
Background: Colonization with methicillin-resistant Staphylococcus aureus(MRSA) poses a hygiene risk that does not spare field hospitals or military medical field camps during military deployments. Diagnostic options ...Background: Colonization with methicillin-resistant Staphylococcus aureus(MRSA) poses a hygiene risk that does not spare field hospitals or military medical field camps during military deployments. Diagnostic options for unambiguously identifying MRSA isolates are usually scarce in military environments. In this study, we assessed the stepwise application of two different selective agars for the specific identification of MRSA in screening analyses.Methods: Nasal swabs from 1,541 volunteers were subjected to thioglycollate broth enrichment and subsequently screened on CHROMagar MRSA selective agar for the identification of MRSA. The MRSA identity of suspiciouslooking colonies was confirmed afterwards or excluded by another selective agar, chrom ID MRSA. All isolates from the selective agars with MRSA-specific colony morphology were identified by biochemical methods and mass spectrometry.Results: The initial CHROMagar MRSA screening identified suspicious colonies in 36 out of 1541 samples. A total of 25 of these 36 isolates showed MRSA-like growth on chrom ID agar. Out of these 25 isolates, 24 were confirmed as MRSA, while one isolate was identified as Staphylococcus kloosii. From the 11 strains that did not show suspicious growth on chrom ID agar, 3 were methicillin-sensitive Staphylococcus aureus(MSSA, with one instance of cocolonization with Corynebacterium spp.), 2 were confirmed as MRSA(with 1 instance of co-colonization with MSSA), 2 were lost during passaging and could not be re-cultured, one could not be identified by the applied approaches, and the remaining 3 strains were identified as Staphylococcus saprophyticus, Staphylococcus hominis(co-colonized with Macrococcus caseolyticus) and Staphylococcus cohnii, respectively.Conclusion: The application of the selective agar CHROMagar MRSA alone proved to be too non-specific to allow for a reliable diagnosis of the presence of MRSA. The combined use of two selective agars in a stepwise approach reduced this non-specificity with an acceptably low loss of sensitivity. Accordingly, such a stepwise screening approach might be an option for resource-restricted military medical field camps.展开更多
Coagulase is considered as a major determinant factor for the identification of Staphylococcus aureus strains.The 3-end coding region of the coagulase(coa)gene contains a series of 81-bp tandem repeats,which differ in...Coagulase is considered as a major determinant factor for the identification of Staphylococcus aureus strains.The 3-end coding region of the coagulase(coa)gene contains a series of 81-bp tandem repeats,which differ in the number and location of enzymatic restriction sites among different isolates.coa PCR-RFLP has been used widely to type S.aureus isolates in epidemiological studies.The current study was conducted to investigate the coagulase gene polymorphisms in S.aureus isolated from various food samples using an in house PCR-RFLP method.A total of 100 strains of S.aureus were isolated from food samples.Isolates were typed by PCR-RFLP analysis using NdeI restriction digestion of the coagulase gene PCR products.Results showed that amplification of coagulase genes from S.aureus produced different PCR products.The isolates were grouped into 18 genotypes using RFLP analysis results of the genes.In this study,the S.aureus isolates have been shown to include more than one coagulase genotype,but only had a few coa genotypes predominated.展开更多
借助扫描电镜和透射电镜对尼罗罗非鱼（Oreochromis niloticus）、奥利亚罗非鱼（O.aureus）、奥尼罗非鱼（O.niloticus×O.aureus）和吉富罗非鱼（GIFT Strain in Nile Tilapia,O.niloticus）成熟精子的超微结构进行了比较研究。...借助扫描电镜和透射电镜对尼罗罗非鱼（Oreochromis niloticus）、奥利亚罗非鱼（O.aureus）、奥尼罗非鱼（O.niloticus×O.aureus）和吉富罗非鱼（GIFT Strain in Nile Tilapia,O.niloticus）成熟精子的超微结构进行了比较研究。结果显示,4种罗非鱼精子均为有鞭毛精子,无顶体,尾部轴丝9±2型。尼罗罗非鱼精子植入窝陷入核深度最浅,线粒体个数和平均直径分别为5-9个、（0.36±0.10）μm,鞭毛侧鳍平均翼长为（0.29±0.11）μm;奥利亚罗非鱼精子核内染色质着色程度最深,植入窝陷入核深度约1/3,线粒体个数和平均直径分别为3-10个、（0.40±0.17）μm,鞭毛侧鳍平均翼长为（0.32±0.12）μm;奥尼罗非鱼精子核内染色质着色程度较浅,植入窝陷入核深度约1/3,线粒体个数和平均直径分别为5-7个、（0.32±0.09）μm,鞭毛侧鳍平均翼长（0.26±0.09）μm;吉富罗非鱼精子核内染色质着色最浅,植入窝陷入核最深,约占2/3,线粒体个数和平均直径分别为5-10个、（0.37±0.08）μm,中段存在大量中空囊泡,鞭毛侧鳍平均翼长为（0.15±0.03）μm,与其它三种罗非鱼精子鞭毛平均侧鳍翼长比较具显著性差异（P〈0.05）。展开更多
基金financially supported by the National Natural Science Foundation of China(82101069,82102537,82160411,82002278)the Natural Science Foundation of Chongqing Science and Technology Commission(CSTC2021JCYJ-MSXMX0170,CSTB2022BSXM-JCX0039)+2 种基金the First Affiliated Hospital of Chongqing Medical University Cultivating Fund(PYJJ2021-02)the Beijing Municipal Science&Technology Commission(Z221100007422130)the Youth Incubation Program of Medical Science and Technology of PLA(21QNPY116).
文摘Background:Treatment of methicillin-resistant Staphylococcus aureus(MRSA)biofilm infections in implant placement surgery is limited by the lack of antimicrobial activity of titanium(Ti)implants.There is a need to explore more effective approaches for the treatment of MRSA biofilm infections.Methods:Herein,an interfacial functionalization strategy is proposed by the integration of mesoporous polydopamine nanoparticles(PDA),nitric oxide(NO)release donor sodium nitroprusside(SNP)and osteogenic growth peptide(OGP)onto Ti implants,denoted as Ti-PDA@SNP-OGP.The physical and chemical properties of Ti-PDA@SNP-OGP were assessed by scanning electron microscopy,X-ray photoelectron spectroscope,water contact angle,photothermal property and NO release behavior.The synergistic antibacterial effect and elimination of the MRSA biofilms were evaluated by 2′,7′-dichlorofluorescein diacetate probe,1-N-phenylnaphthylamine assay,adenosine triphosphate intensity,O-nitrophenyl-β-D-galactopyranoside hydrolysis activity,bicinchoninic acid leakage.Fluorescence staining,assays for alkaline phosphatase activity,collagen secretion and extracellular matrix mineralization,quantitative real‑time reverse transcription‑polymerase chain reaction,and enzyme-linked immunosorbent assay(ELISA)were used to evaluate the inflammatory response and osteogenic ability in bone marrow stromal cells(MSCs),RAW264.7 cells and their co-culture system.Giemsa staining,ELISA,micro-CT,hematoxylin and eosin,Masson's trichrome and immunohistochemistry staining were used to evaluate the eradication of MRSA biofilms,inhibition of inflammatory response,and promotion of osseointegration of Ti-PDA@SNP-OGP in vivo.Results:Ti-PDA@SNP-OGP displayed a synergistic photothermal and NO-dependent antibacterial effect against MRSA following near-infrared light(NIR)irradiation,and effectively eliminated the formed MRSA biofilms by inducing reactive oxygen species(ROS)-mediated oxidative stress,destroying bacterial membrane integrity and causing leakage of intracellular components(P<0.01).In vitro experiments revealed that Ti-PDA@SNP-OGP not only facilitated osteogenic differentiation of MSCs,but also promoted the polarization of pro-inflammatory M1 macrophages to the anti-inflammatory M2-phenotype(P<0.05 or P<0.01).The favorable osteo-immune microenvironment further facilitated osteogenesis of MSCs and the anti-inflammation of RAW264.7 cells via multiple paracrine signaling pathways(P<0.01).In vivo evaluation confirmed the aforementioned results and revealed that Ti-PDA@SNP-OGP induced ameliorative osseointegration in an MRSA-infected femoral defect implantation model(P<0.01).Conclusions:Ti-PDA@SNP-OGP is a promising multi-functional material for the high-efficient treatment of MRSA infections in implant replacement surgeries.
基金supported by Brain Korea (BK)21 Plus Project (4299990913942)funded by the Korean Government,Koreathe Collabo Project funded by the Ministry of SMEs and Startups (C1016120-01-02)the National Research Foundation of Korea (NRF) (2018007551)。
文摘Slightly acidic electrolyzed water(SAEW)has proven to be an efficient and novel sanitizer in food and agriculture field.This study assessed the efficacy of SAEW(30 mg/L)at 40℃on the inactivation of foodbome pathogens and detachment of multi-resistant Staphylococcus aureus(MRSA)biofilm.Furthermore.the underlying mechanism of MRS A biofilm under heated SAEW at 40℃treatment on metabolic profiles was investigated.The results showed that the heated SAEW at 40℃significantly effectively against foodbome pathogens of 1.96-7.56(lg(CFU/g))reduction in pork,chicken,spinach,and lettuce.The heated SAEW at 40℃treatment significantly reduced MRS A biofilm cells by 2.41(lg(CFU/cm^(2))).The synergistic effect of SAEW treatment showed intense anti-biofilm activity in decreasing cell density and impairing biofilm cell membranes.Global metabolic response of MRSA biofilms,treated by SAEW at 40℃,revealed the alterations of intracellular metabolites,including amino acids,organic acid,fatty acid,and lipid.Moreover,signaling pathways involved in amino acid metabolism,energy metabolism,nucleotide synthesis,carbohydrate metabolites,and lipid biosynthesis were functionally disrupted by the SAEW at 40℃treatment.As per our knowledge,this is the first research to uncover the potential mechanism of heated SAEW treatment against MRSA biofilm on food contact surface.
基金supported by the National Natural Science Foundation of China (31972169 and 32001798)。
文摘Staphylococcus aureus is a serious foodborne pathogen threatening food safety and public health.Especially the emergence of methicillin-resistant Staphylococcus aureus(MRSA)increased the difficulty of S.aureus treatment.Staphyloxanthin is a crucial virulence factor of S.aureus.Blocking staphyloxanthin production could help the host immune system counteract the invading S.aureus cells.In this study,we first screened for staphyloxanthin inhibitors using a virtual screening method.The outcome of the virtual screening method resulted in the identification of eugenol(300μg/mL),which significantly inhibits the staphyloxanthin production in S.aureus ATCC 29213,S.aureus Newman,MRSA ATCC 43300 and MRSA ATCC BAA1717by 84.2%,63.5%,68.1%,and 79.5%,respectively.The outcome of the growth curve assay,field-emission scanning electron,and confocal laser scanning microscopy analyses confirmed that eugenol at the test concentration did not affect the morphology and growth of S.aureus.Moreover,the survival rate of S.aureus ATCC 29213 and MRSA ATCC 43300 under H_(2)O_(2) pressure decreased to 51.9%and 45.5%in the presence of eugenol,respectively.The quantitative RT-PCR and molecular simulation studies revealed that eugenol targets staphyloxanthin biosynthesis by downregulating the transcription of the crtM gene and inhibiting the activity of the CrtM enzyme.Taken together,we first determined that eugenol was a prominent compound for staphyloxanthin inhibitor to combat S.aureus especially MRSA infections.
基金This work was financially supported by Major Science and Technology Project of Yunnan Province(202302AE090022)Key Research and Development Program of Yunnan(202203AC100010)+4 种基金the National Natural Science Foundation of China(32160597,32160236,32371463)National Key Research and Development Program of China(2022YFC2601604)Cardiovascular Ultrasound Innovation Team of Yunnan Province(202305AS350021)Spring City Plan:the High-level Talent Promotion and Training Project of Kunming(2022SCP001)the second phase of“Double-First Class”Program Construction of Yunnan University.
文摘Sensitive detection of Staphylococcus aureus enterotoxin B(SEB)is of importance for preventing food poisoning from threatening human health.In this work,an electrochemical and colorimetric dual-signal detection assay of SEB was developed.The probe(Ab2/AuPt@Fe-N-C)was bound to SEB captured by Ab1,where the Ab2/AuPt@Fe-N-C triggered methylene blue degradation and resulted in the decrease of electrochemical signal.Furthermore,the probe catalyzed the oxidation of 3,3’,5,5’-tetramethyl biphenyl to generate a colorimetric absorbance at 652 nm.Once the target was captured and formed a sandwich-like complex,the color changed from colorless to blue.SEB detection by colorimetric and electrochemical methods showed a linear relationship in the concentration ranges of 0.0002-10.0000 and 0.0005-10.0000 ng/mL,with limits of detection of 0.0667 and 0.1670 pg/mL,respectively.The dual-signal biosensor was successfully used to detect SEB in milk and water samples,which has great potential in toxin detection in food and the environment.
基金Supported by the Doctor Research Start-up Fund of Liaoning province (20081055)a grant from the Education Department of Liaoning province (2009A737)
文摘Objective To explore the mechanisms involved in Staphylococcus aureus （S. aureus） invading human monocytic U937 cells. Methods S. aureus were added to U937 cells at multiplicity of infections （MOI） of 20：1 for 0, 15, 30, 60, and 90 minutes, respectively. Cell apoptosis was analyzed with Hoechst 33258 staining and Annexin V-fluorescein isothiocyanate （FITC）/propidium iodide （PI） flow cytometry analysis. Akt and nuclear factor-κB （NF-κB） activities were detected by Western blotting. Results Infection of U937 cells with S. aureus induced rapid cell death in a time-dependent manner, and the cells displayed characteristic features of apoptosis. S. aureus-induced apoptosis was associated with a prominent downregulation of activated （phosphorylated） Akt and NF-κB. The inhibition of phosphorylated Akt by LY294002 led to the inhibition of NF-κB in a dose-dependent manner. Inhibition of Akt with LY294002 caused further increase in apoptosis of U937 cells. Conclusions S. aureus can stimulate the apoptosis of U937 ceils. S. aureus induces apoptosis of U937 cells by inhibiting Akt-regulated NF-κB.
基金Supported by the Doctor Research Start-up Fund of Liaoning province (20081055) a grant from the Education Department of Liaoning province (2008771)
文摘Objective To investigate the effect of SP600125, a specific c-jun N-terminal protein kinase (JNK) inhibitor, on Staphylococcus aureus (S. aureus)-induced U937 cell death and the underlying mechanism. Methods The human monocytic U937 cells were treated with S. aureus at different time with or without SP600125. Cell apoptosis was analyzed by flow cytometry. JNK, Bax, and caspase-3 activities were detected by Western blotting. Results S. aureus induced apoptosis in cultured U937 cells in a time-dependent manner. Expression of Bax and phospho-JNK significantly increased in S. aureus-treated U937 cells, and the level of activated caspase-3 also increased in a time-dependent manner. Inhibition of JNK with SP600125 significantly inhibited S. aureus-induced apoptosis in U937 cells. Conclusions S. aureus can induce apoptosis in U937 cells by phosphorylation of JNK and activation of Bax and caspase-3. SP600125 protects U937 cells from apoptosis induced by S. aureus via inhibiting the activity of JNK.
基金the National Major R&D Program of China(grant 2016YFD0400301)the National Natural Science Foundation of China(grants 31401608).
文摘Ultrasound,is thought to a potential non-thermal sterilization technology in food industry.However,the exact mechanisms underlying microbial inactivation by ultrasound still remain obscure.In this study,the action modes of ultrasound on both Gram-negative and Gram-positive microorganisms were estimated.From colony results,ultrasound acted as an irreversible effect on both Eshcerichia coli and Staphylococcus aureus without sublethal injury.The result in this study also showed that a proportion of bacteria subpopulation suffered from serious damage of intracellular components(e.g.DNA and enzymes)but with intact cell envelopes.We speculated that the inactivated effects of ultrasound on microbes might more than simply completed disruption of cell exteriors.Those microbial cells who had not enter the valid area of ultrasonic cavitation might be injected with free radicals produced by ultrasound and experienced interior injury with intact exterior structure,and others who were in close proximity to the ultrasonic wave field would be immediately and completely disrupted into debris by high power mechanic forces.These findings here try to provide extension for the inactivation mechanisms of ultrasound on microorganisms.
基金Development of Application Technology Project(No:2015-114)issued by Science and Technology Committee of Shanghai Municipal GovernmentNational Key Scientific Instruments Project(No:2013YQ150557)issued by Ministry of Science and Technology of the P.R.China.
文摘This study was conducted to analyze the effects of sodium nitrite,nisin,potassium sorbate,and sodium lactate against Staphylococcus aureus(S.aureus)growth and staphylococcal enterotoxins(SEs)production in cooked pork sausage by inoculating sausage samples containing preservative with an S.aureus strain producing staphylococcal enterotoxin A(SEA)and then storing them at 37℃ for 36 h.Samples were analyzed every 3 h to count the S.aureus colonies and to detect SEA.The modified Gompertz model was used to describe S.aureus growth in the samples under various conditions,and the preservatives with a significant antimicrobial effect were selected.In addition,the antimicrobial effects of the selected preservatives under various concentrations were tested.Results showed that sodium nitrite,nisin,and potassium sorbate had a weak effect against S.aureus growth and had no effect against SEA production,whereas sodium lactate could significantly inhibit S.aureus growth and SEA production.Moreover,the antimicrobial effect of sodium lactate was concentration-dependent,wherein sodium lactate concentration<12 g/kg showed no inhibitory effect,but when the concentration was increased to 24 g/kg,sodium lactate could effectively inhibit S.aureus growth and SEA production,and at 48 g/kg,sodium lactate had a significant inhibitory effect.
基金The research leading to these results is partly funded by the research agreement between Istituto di Fisica del Plasma and Kenosistec Srl(protocol CNR-IFP No.63,22/01/2018).
文摘In this paper,a low-pressure capacitively coupled plasma discharge sustained in an argonoxygen mixture was studied in order to evaluate its properties in terms of inactivation of Staphylococcus aureus.The plasma parameters as electron temperature and plasma density were measured by the Langmuir probe(Ne≈1015 m^-3,Te≈1.5 eV),while the neutral atom density was in the range of 1021 m^-3.In the plasma phase,oxygen radicals were taken as reference of the reactive species with antimicrobial activity,and oxygen spectral lines,over a range of plasma process parameters,were investigated by the optical emission spectroscopy.Optimal plasma conditions were found,and a count reduction of 4 log in a few minutes of the bacterium proves the potentiality of an industrial grade plasma reactor as a sterilization agent.
文摘BACKGROUND:Over the past two decades,methicillin-resistant Staphylococcus aureus(MRSA) has evolved from a hospital-associated infection to a significant public health threat in the community,causing outbreaks of soft tissue infections in otherwise healthy individuals.The goal of this study was to determine the prevalence of nasal MRSA colonization in low acuity Emergency Department(ED) Fast Track patients in order to better characterize the epidemiology of this pathogen.METHODS:We conducted a cross-sectional study of a convenience sample of adult patients from our ED Fast Track.Nasal swabs were analyzed for MRSA using a polymerase chain reaction assay.Study participants completed a survey assessing traditional risk factors for CA-MRSA colonization.RESULTS:A total of 106 ED Fast Track patients were tested.Four(3.8%,95%CI 1.5%-9.3%)were MRSA positive.Three traditional CA-MRSA risk factors(personal history of abscess,family history of abscess,and participation in contact sports) were examined.In patients with a positive MRSA nasal swab,only a personal prior history of abscess retained significance(OR 33,95%C11.7-676,P=0.02).CONCLUSION:This study found a higher prevalence of nasal MRSA colonization in low acuity ED Fast Track patients compared with historical community surveillance studies.A personal history of prior abscess was a significant risk for CA-MRSA carriage.
文摘Background: Colonization with methicillin-resistant Staphylococcus aureus(MRSA) poses a hygiene risk that does not spare field hospitals or military medical field camps during military deployments. Diagnostic options for unambiguously identifying MRSA isolates are usually scarce in military environments. In this study, we assessed the stepwise application of two different selective agars for the specific identification of MRSA in screening analyses.Methods: Nasal swabs from 1,541 volunteers were subjected to thioglycollate broth enrichment and subsequently screened on CHROMagar MRSA selective agar for the identification of MRSA. The MRSA identity of suspiciouslooking colonies was confirmed afterwards or excluded by another selective agar, chrom ID MRSA. All isolates from the selective agars with MRSA-specific colony morphology were identified by biochemical methods and mass spectrometry.Results: The initial CHROMagar MRSA screening identified suspicious colonies in 36 out of 1541 samples. A total of 25 of these 36 isolates showed MRSA-like growth on chrom ID agar. Out of these 25 isolates, 24 were confirmed as MRSA, while one isolate was identified as Staphylococcus kloosii. From the 11 strains that did not show suspicious growth on chrom ID agar, 3 were methicillin-sensitive Staphylococcus aureus(MSSA, with one instance of cocolonization with Corynebacterium spp.), 2 were confirmed as MRSA(with 1 instance of co-colonization with MSSA), 2 were lost during passaging and could not be re-cultured, one could not be identified by the applied approaches, and the remaining 3 strains were identified as Staphylococcus saprophyticus, Staphylococcus hominis(co-colonized with Macrococcus caseolyticus) and Staphylococcus cohnii, respectively.Conclusion: The application of the selective agar CHROMagar MRSA alone proved to be too non-specific to allow for a reliable diagnosis of the presence of MRSA. The combined use of two selective agars in a stepwise approach reduced this non-specificity with an acceptably low loss of sensitivity. Accordingly, such a stepwise screening approach might be an option for resource-restricted military medical field camps.
文摘Coagulase is considered as a major determinant factor for the identification of Staphylococcus aureus strains.The 3-end coding region of the coagulase(coa)gene contains a series of 81-bp tandem repeats,which differ in the number and location of enzymatic restriction sites among different isolates.coa PCR-RFLP has been used widely to type S.aureus isolates in epidemiological studies.The current study was conducted to investigate the coagulase gene polymorphisms in S.aureus isolated from various food samples using an in house PCR-RFLP method.A total of 100 strains of S.aureus were isolated from food samples.Isolates were typed by PCR-RFLP analysis using NdeI restriction digestion of the coagulase gene PCR products.Results showed that amplification of coagulase genes from S.aureus produced different PCR products.The isolates were grouped into 18 genotypes using RFLP analysis results of the genes.In this study,the S.aureus isolates have been shown to include more than one coagulase genotype,but only had a few coa genotypes predominated.
文摘借助扫描电镜和透射电镜对尼罗罗非鱼（Oreochromis niloticus）、奥利亚罗非鱼（O.aureus）、奥尼罗非鱼（O.niloticus×O.aureus）和吉富罗非鱼（GIFT Strain in Nile Tilapia,O.niloticus）成熟精子的超微结构进行了比较研究。结果显示,4种罗非鱼精子均为有鞭毛精子,无顶体,尾部轴丝9±2型。尼罗罗非鱼精子植入窝陷入核深度最浅,线粒体个数和平均直径分别为5-9个、（0.36±0.10）μm,鞭毛侧鳍平均翼长为（0.29±0.11）μm;奥利亚罗非鱼精子核内染色质着色程度最深,植入窝陷入核深度约1/3,线粒体个数和平均直径分别为3-10个、（0.40±0.17）μm,鞭毛侧鳍平均翼长为（0.32±0.12）μm;奥尼罗非鱼精子核内染色质着色程度较浅,植入窝陷入核深度约1/3,线粒体个数和平均直径分别为5-7个、（0.32±0.09）μm,鞭毛侧鳍平均翼长（0.26±0.09）μm;吉富罗非鱼精子核内染色质着色最浅,植入窝陷入核最深,约占2/3,线粒体个数和平均直径分别为5-10个、（0.37±0.08）μm,中段存在大量中空囊泡,鞭毛侧鳍平均翼长为（0.15±0.03）μm,与其它三种罗非鱼精子鞭毛平均侧鳍翼长比较具显著性差异（P〈0.05）。
