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1.
A great amount of foodborne pathogens were Gram-positive(G+) bacteria, a threat to public health. In this study, considering the binding ability of nisin towards G+ bacteria and the stable fluorescent ability of EGFP protein, a fluorescent nisin–EGFP protein probe was constructed by a gene engineering method. Nisin and EGFP were used as the receptor and fluorophore, respectively, to detect G+ bacteria. The nisin and egfp gene were amplified separately according to the sequence published in Gen Bank using unique primers. The two genes were cloned into a pET-28b(+) vector resulting in apET-28b(+)–nisin–egfp vector. The vector was transferred into Escherichia coli(E. coli) BL21(DE3) for expression. The expressed protein was extracted, purified by a Ni–NTA column, and then tested by the SDS-PAGE method to confirm its molecular weight. Listeria monocytogenes(L.monocytogenes), Staphylococcus aureus(S. aureus), and Micrococcus luteus(M. luteus) were used as the representations of G+ bacteria. E. coli O157, representing the gram-negative(G-) bacteria, was used as a negative control. The binding specificity of the recombinant protein was performed on two types of bacteria and then detected through fluorescent microscopy. The results indicated that the nisin–EGFP probe could detect G+ bacteria at 10~8CFU/mL.  相似文献   

2.
Six lactic acid bacterial (LAB) strains were isolated from traditionally fermented Xinjiang cheese and evaluated for functional and probiotic properties and potentials as starter cultures. The isolated six LAB strains comprised Lactobacillus rhamnosus (one strain), Lactobacillus helveticus (one strain), and Enterococcus hirae (four strains). All of the six strains were tolerant to acidic and bile salt conditions. Among which, the L. rhamnosus R4 strain showed more desirable antimicrobial, auto-aggregation, and hydrophobic activity. In addition, the strain L. rhamnosus R4 exhibited the highest level of free radical scavenging activity (53.78% of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals and 45.79% of hydroxyl radicals). L. rhamnosus R4 also demonstrated cholesterol and triglyceride degradation by 50.97% and 28.92%, respectively. To further examine the health-promoting effects of these LAB strains on host lifespan, Caenorhabditis elegans was used as an in vivo model. Worms fed LAB as a food source had significant differences in lifespan compared to those fed Escherichia coli OP50 (as a negative control). Feeding of L. rhamnosus R4 extended the mean lifespan of C. elegans by up to 36.1% compared to that of the control. The results suggest that the strains isolated from Xinjiang fermented dairy products have high potential as starter cultures in the cheese industry.  相似文献   

3.

Objective

To investigate the antibacterial mechanism of high-mobility group nucleosomal-binding domain 2 (HMGN2) on Escherichia coli K12, focusing on the antibacterial and antibiofilm formation effects. Its chemotactic activity on human neutrophils was also investigated.

Methods

Human tissue-derived HMGN2 (tHMGN2) was extracted from fresh uterus fiber cystadenoma and purified by HP1100 reversed-phase high-performance liquid chromatography (RP-HPLC). Recombinant human HMGN2 (rHMGN2) was generated in E. coli DE3 carrying PET-32ac(+)- HMGN2. Antibacterial activity of HMGN2 was determined using an agarose diffusion assay and minimum inhibitory concentration (MIC) of HMGN2 was determined by the microdilution broth method. Bacterial membrane permeability assay and DNA binding assay were performed. The antibiofilm effect of HMGN2 was investigated using a crystal violet assay and electron microscopy scanning. The activating effect and chemotactic activity of HMGN2 on neutrophils were determined using a nitroblue tetrazolium (NBT) reduction assay and Transwell chamber cell migration assay, respectively.

Results

HMGN2 showed a relatively high potency against Gram-negative bacteria E. coli and the MIC of HMGN2 was 16.25 μg/ml. Elevated bacterial membrane permeability was observed in HMGN2-treated E. coli K12. HMGN2 could also bind the bacterial plasmid and genomic DNA in a dose-dependent manner. The antibiofilm effect of HMGN2 on E. coli K12 was confirmed by crystal violet staining and scanning electron microscopy. However, the activating effects and chemotactic effects of HMGN2 on human neutrophils were not observed.

Conclusions

As an antimicrobial peptide (AMP), HMGN2 possessed a good capacity for antibacterial and antibiofilm activities on E. coli K12. This capacity might be associated with disruption of the bacterial membrane and combination of DNA, which might affect the growth and viability of E. coli.
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4.
The Bacillus strain BH072 isolated from a honey sample showed strong antifungal activity against phytopathogen. Gene cloning test demonstrated that the strain had a tasA gene encoding an antifungal TasA protein. Although the wild strain simultaneously produced various antifungal substances, only the physicochemical property and antifungal activity of TasA protein were unclear due to the difficulty in extraction. In this study, tasA gene encoding the protein from Bacillus sp. BH072 was amplified by using the polymerase chain reaction (PCR) method and cloned into pET 28a (+) vector, and then expressed in host cells Escherichia coli BL21 (DE3). The expressed proteins were collected by centrifugation and ultrasonic treatment, and then purified by using nickel-nitrilotriacetic acid (Ni-NTA) metal affinity column and dialysis methods. The result of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test showed that an expected protein band appeared with a size of 31 kDa. The expressed products possessed antifungal activity against the phytopathogenic indicator strain Botrytis cinerea. A genetically engineered strain tasA of E. coli was established in this study which can efficiently express Tas A protein.  相似文献   

5.
In this study,we aimed to examine the inhibitory effect of PA003,a Pediococcus acidilactici that produces lactic acid and antimicrobial peptides pediocin,on pathogenic biofilm formation on abiotic surfaces.PA003 and pathogens(Escherichia coli,Salmonella enterica serovar Typhimurium,Staphylococcus aureus and Listeria monocytogenes) were used to evaluate auto-aggregation,hydrophobicity,biofilm formation and biofilm formation inhibition on stainless steel,polyvinyl chloride and glass slides in terms of exclusion,displacement and competition.The results showed the highest auto-aggregation abilities were observed for one of the E.coli strains EAggEC(E58595) and the highest hydrophobic strain was observed with EPEC(E2348/69)(51.9%).The numbers of biofilm cells of E.coli,S.Typhimurium,S.aureus and L.monocytogenes on stainless steel,polyvinyl chloride and glass slide coupons were effectively reduced by approximately 4log CFU/coupon.These results demonstrate that lactic acid bacteria can be used as an alternative to effectively control the formation of biofilms by food-borne pathogens.  相似文献   

6.
Children from low-income families are at greater risk for poor social–emotional development and physical health and may be in need of intervention. This study examined the extent to which the Positive Action (PA) preschool lessons improved low-income children’s social–emotional competence and health behaviors. Mixed findings emerged with regard to whether the lessons facilitated growth in child outcomes. Results showed positive effects of PA on children’s directly assessed social problem solving skills and their parent-rated social–emotional competence and health behaviors. Unexpectedly, program effects on teacher ratings of social–emotional competence were in the opposite direction. These findings provide preliminary evidence that PA may be effective in promoting positive social–emotional and health behavior outcomes for children, particularly when outcomes are assessed directly and by parents.  相似文献   

7.
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8.

Objective

Salmonella enterica remains a major cause of food-borne disease in humans, and Salmonella Typhimurium (ST) contamination of poultry products is a worldwide problem. Since macrophages play an essential role in controlling Salmonella infection, the aim of this study was to evaluate the effect of glycyrrhizic acid (GA) on immune function of chicken HD11 macrophages.

Methods

Chicken HD11 macrophages were treated with GA (0, 12.5, 25, 50, 100, 200, 400, or 800 μg/ml) and lipopolysaccharide (LPS, 500 ng/ml) for 3, 6, 12, 24, or 48 h. Evaluated responses included phagocytosis, bacteria-killing, gene expression of cell surface molecules (cluster of differentiation 40 (CD40), CD80, CD83, and CD197) and antimicrobial effectors (inducible nitric oxide synthase (iNOS), NADPH oxidase-1 (NOX-1), interferon-γ (IFN-γ), LPS-induced tumor necrosis factor (TNF)-α factor (LITAF), interleukin-6 (IL-6), and IL-10), and production of nitric oxide (NO) and hydrogen peroxide (H2O2).

Results

GA increased the internalization of both fluorescein isothiocyanate (FITC)-dextran and ST by HD11 cells and markedly decreased the intracellular survival of ST. We found that the messenger RNA (mRNA) expression of cell surface molecules (CD40, CD80, CD83, and CD197) and cytokines (IFN-γ, IL-6, and IL-10) of HD11 cells was up-regulated following GA exposure. The expression of iNOS and NOX-1 was induced by GA and thereby the productions of NO and H2O2 in HD11 cells were enhanced. Notably, it was verified that nuclear factor-κB (NF-κB) and c-Jun N-terminal kinase (JNK) pathways were responsible for GA-induced synthesis of NO and IFN-γ gene expression.

Conclusions

Taken together, these results suggested that GA exhibits a potent immune regulatory effect to activate chicken macrophages and enhances Salmonella-killing capacity.
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9.
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10.
In plants, lipoxygenases (LOXs) play a crucial role in biotic and abiotic stresses. In our previous study, five 13-LOX genes of oriental melon were regulated by abiotic stress but it is unclear whether the 9-LOX is involved in biotic and abiotic stresses. The promoter analysis revealed that CmLOX09 (type of 9-LOX) has hormone elements, signal substances, and stress elements. We analyzed the expression of CmLOX09 and its downstream genes—CmHPL and CmAOS—in the leaves of four-leaf stage seedlings of the oriental melon cultivar “Yumeiren” under wound, hormone, and signal substances. CmLOX09, CmHPL, and CmAOS were all induced by wounding. CmLOX09 was induced by auxin (indole acetic acid, IAA) and gibberellins (GA3); however, CmHPL and CmAOS showed differential responses to IAA and GA3. CmLOX09, CmHPL, and CmAOS were all induced by hydrogen peroxide (H2O2) and methyl jasmonate (MeJA), while being inhibited by abscisic acid (ABA) and salicylic acid (SA). CmLOX09, CmHPL, and CmAOS were all induced by the powdery mildew pathogen Podosphaera xanthii. The content of 2-hexynol and 2-hexenal in leaves after MeJA treatment was significantly higher than that in the control. After infection with P. xanthii, the diseased leaves of the oriental melon were divided into four levels—levels 1, 2, 3, and 4. The content of jasmonic acid (JA) in the leaves of levels 1 and 3 was significantly higher than that in the level 0 leaves. In summary, the results suggested that CmLOX09 might play a positive role in the response to MeJA through the hydroperoxide lyase (HPL) pathway to produce C6 alcohols and aldehydes, and in the response to P. xanthii through the allene oxide synthase (AOS) pathway to form JA.  相似文献   

11.
Tomato is an ideal model species for fleshy fruit development research. SlYABBY2b regulates the ovary locule number, which is increased by gibberellins, in tomato. However, the relationship between SlYABBY2b and endogenous gibberellin is poorly understood. In this study, SlYABBY2b-overexpressing and RNA interference (RNAi) transgenic tomato plants were used to elucidate the mechanism by which SlYABBY2b regulates the ovary locule number and endogenous gibberellin content in tomato. SlYABBY2b-overexpressing plants showed fewer locules and lower gibberellin content than the control plants. Contrasting results were found in the RNAi lines. Therefore, the SlYABBY2b gene negatively regulates tomato ovary locule number and endogenous gibberellin content. Furthermore, the expression of SlYABBY2b gene was remarkably higher than that of the wild type in the apical shoots of gibberellindeficient mutants. This showed that the gibberellins can inhibit the expression of SlYABBY2b gene negative regulation. Further study revealed that SlYABBY2b suppressed the expression of SlGA20ox1 and SlGA3ox2, but increased that of SlGA2ox1 and SlGA2ox5 in the apical shoots of SlYABBY2b-overexpressing plants, thereby reducing gibberellin content. Contrasting results were found in the RNAi lines. Our results showed that the SlYABBY2b gene was located on gibberellin signal transduction pathways, fed back regulation of the synthesis of gibberellin, and felt exogenous gibberellin signal to further regulate the formation of tomato locule.  相似文献   

12.
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13.
This article examines the productivity of both individuals and institutions, indexed through an examination of five educational psychology journals (Cognition and Instruction, Contemporary Educational Psychology, Educational Psychologist, Educational Psychology Review, and Journal of Educational Psychology) from 2009 to 2014. These results are discussed relative to four previous studies (Hsieh et al. Contemporary Educational Psychology, 29, 333–343, 2004; Jones et al. Contemporary Educational Psychology, 35, 11–16, 2010; Smith et al. Contemporary Educational Psychology, 23, 173–181, 1998; Contemporary Educational Psychology, 28, 422–430, 2003). Vanderbilt University and Fred Paas replaced the University of Maryland and Richard E. Mayer as the top research institution and author, respectively. Sixteen of the top 19 researchers’ institutions were outside the USA, compared to only 10 of the top 32 during 2003–2008 and three of the top 20 during 1991–1996. Educational psychology research continues the trend of becoming more international.  相似文献   

14.
The objective of this study was to describe the fecal microbiota succession of piglets from birth to post-weaning by pyrosequencing. The dominant bacterial phyla were Bacteroidetes, Firmicutes, Proteobacteria, and Fusobacteria, although the composition of the fecal microbiota changed as the piglets grew. Bacteroides, Parabacteroides, Prevotella, Lactobacillus, unclassified Lachnospiraceae, Ruminococcus, Oscillospira, Phascolarctobacterium, and Desulfovibrio were the core genera in the feces of piglets and existed from birth to post-weaning. Bacteroides was the most abundant at birth; however, the proportion of Bacteroides decreased as the piglets aged, while the proportion of Prevotella increased until it became the most abundant genus at the post-weaning stage. Principal coordinate analysis indicated that the microbial communities of piglet fecal samples from birth to post-weaning could be separated into four groups according to the similarity of community composition. These corresponded to time points on day 0 (birth); day 7; days 14, 21, and 28 (pre-weaning); and day 35 (post-weaning). This study suggests that the development of the intestinal microbiota in the piglet is a gradual and sequential process. This methodology establishes a basis for broader studies to investigate the microbiota of the piglet intestinal tract.  相似文献   

15.
Chlamydia psittaci is an important zoonotic pathogen in birds and may be transmitted to humans and result in severe respiratory disease. To assess the prevalence and genotype of C. psittaci in birds in Kunming, Yunnan, China, a total of 136 specimens of psittacine birds involving 8 species were collected from the city’s zoos (n=60) and pet markets (n=76). The frequency of C. psittaci infection was 19.9% (27/136) in the psittacine birds. The prevalence of C. psittaci was higher in pet birds (26.3%; 20/76) than in zoo birds (11.7%; 7/60) (P=0.034). In particular, among Agapornis fischeri, the C. psittaci infection (50%; 10/20) was significantly more frequent in the pet markets than in the zoos (P=0.006). In addition, the highest prevalence of 41.2% (7/17) was found in Ara ararauna. To determine the genotype of C. psittaci, 23 OmpA gene fragments (about 1.4 kb) in 27 positive samples were successfully amplified and sequenced. Phylogenetic analysis showed that all the 23 strains belonged to genotype A. Our results demonstrate the high prevalence of C. psittaci genotype A infection in psittacine birds in Yunnan Province, suggesting a potential threat to human health in this area. Therefore, it is necessary to take effective measures to prevent the spread of C. psittaci among psittacine birds, as well as among employees and customers.  相似文献   

16.
Microbial attacks during storage are one of the primary causes of product deterioration, and can limit the process of prolonging the shelf-life of harvested food. In this study, sweet potatoes were stored at temperatures of 13, 21, and 29 °C for 4 weeks. Samples were collected during storage and plated on potato dextrose agar, from which axenic mold cultures were obtained and identified using 26S rRNA gene sequences. Physiological changes of potato tubers were assessed with respect to pathogenicity, enzyme activity, and atmospheric storage conditions. Six fungal species were identified, namely Penicillium chrysogenum (P. rubens), P. brevicompactum, Mucor circinelloides, Cladosporium cladosporiodes, P. expansum, and P. crustosum. The following fungal isolates, namely P. expansum, P. brevicompactum, and Rhizopus oryzae, were recovered from the re-infected samples and selected according to their levels of enzyme activity. This study revealed high levels of activity for cellulase and pectinase, which were most notable during the initial three days of testing, and were followed by a steady decrease (P<0.05). Polygalacturonase activity was prominent with values ranging from 12.64 to 56.79 U/mg (P. expansum) and 18.36 to 79.01 U/mg (P. brevicompactum). Spoilage was obvious in the control group, which had a 100% decay at the end of the experimental period compared with samples treated with iprodione and sodium hypochlorite, in which the decay rates were 5% and 55%, respectively. The data for the iprodione- and sodium hypochlorite-treated samples at the end of the 3-month storage period showed that they were significantly different (P=0.041), with the sodium hypochlorite-treated samples producing twice the rate of infection compared to the iprodione-treated samples. The comparative rate of the progression of decay in the treated samples can be expressed as iprodione<sodium hypochlorite<control. This study demonstrates that sweet potato tissue damage is due to the activities of microbial enzymes and, in particular, the pectinases of the organisms isolated from the infected potato tissues, and suggests the advantages of utilizing iprodione as a curing agent for potato tubers before storage.  相似文献   

17.
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18.
1　Introduction　AmatrixA∈Cn×missaidtobecentrohermitianifA =Jn AJm,andskewcentrohermitianifA =-Jn AJm,where Adenotetheconjugateofthema trixAandJlisthepermutationmatrixoforderlwithonesonthecrossdiagonal(bottomlefttotopright)andzeroselsewhere .ThehermitianToeplitzmatricesformanimportantsubclassofcentrohermi tianmatriceswhichoccurnaturallyindigitalsignalprocessingandotherareas ,seeforinstance[1] andreferencetherein .Thecentrohermitianandrelatedmatricesrecentlyplayanimportantroleinthesolu…  相似文献   

19.
With the availability of the whole genome sequence of Escherichia coli or Corynebacterium glutamicum, strategies for directed DNA manipulation have developed rapidly. DNA manipulation plays an important role in understanding the function of genes and in constructing novel engineering bacteria according to requirement. DNA manipulation involves modifying the autologous genes and expressing the heterogenous genes. Two alternative approaches, using electroporation linear DNA or recombinant suicide plasmid, allow a wide variety of DNA manipulation. However, the over-expression of the desired gene is generally executed via plasmid-mediation. The current review summarizes the common strategies used for genetically modifying E. coli and C. glutamicum genomes, and discusses the technical problem of multi-layered DNA manipulation. Strategies for gene over-expression via integrating into genome are proposed. This review is intended to be an accessible introduction to DNA manipulation within the bacterial genome for novices and a source of the latest experimental information for experienced investigators.  相似文献   

20.
This empirically driven paper is about workplace learning with specific focus on the ‘work’ of consuming practices. By consuming we refer to the eating, and the drinking, and (at times) to the smoking that workers, in most organisations, do on a daily basis. Indeed, it is the quotidian nature of consuming, coupled with its absence from workplace learning research that make them noteworthy practices to explore. In using the term practice we draw on the recent tranche of practice based theorisations: notably Schatzki (1996, Organization Studies, 26(3), 465-484, 2005, Organization Studies, 27(12), 1863-1873, 2006) and Gherardi (Human Relations, 54(1), 131-139, 2001, 2006, Learning Organization, 16(5), 352-359, 2009). The paper frames consuming practices as ‘dispersed’ (general) practices and, illustrated through empirical data from multiple projects, we progressively outline how these contribute to the learning of ‘integrative’ (specialized work) practices. Our overall aim is to (re)position consuming practices from prosaic, to having much relevance for research on workplace learning.  相似文献   

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