Cloning and efficient expression of <Emphasis Type="Italic">Bacillus sp.</Emphasis> BH072 <Emphasis Type="Italic">tasA</Emphasis> gene in <Emphasis Type="Italic">Escherichia coli</Emphasis>

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.     

<Emphasis Type="Italic">Bildung</Emphasis> in globalizing times

This key paper is intended to produce a comparison of Bildung and globalization, acknowledging that Bildung and globalization are both fuzzy concepts. Our comparison of globalization and Bildung therefore is to be seen as a construction. To reach this objective, we start with the identification of dimensions of the concept of globalization based predominantly on sociological research. This analysis is followed by explication of dimensions of the concept of Bildung as they can be identified before a frame of globalization. We want to find out whether it makes sense to see globalization as one part of a new model of Bildung. In the conclusion, we demonstrate the normativity of each and any theory of Bildung in globalizing times, and we explain why it is necessary to reflect on the normativity of the different concepts. We indicate where we see connections with the other papers of this special issue of “Zeitschrift für Erziehungswissenschaft”.     

Statistical culture-based strategies to enhance chlamydospore production by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> SH2303 in liquid fermentation

Trichoderma-based formulations are applied as commercial biocontrol agents for soil-borne plant pathogens. Chlamydospores are active propagules in Trichoderma spp., but their production is currently limited due to a lack of optimal liquid fermentation technology. In this study, we explored response surface methodologies for optimizing fermentation technology in Trichoderma SH2303. Our initial studies, using the Plackett-Burman design, identified cornmeal, glycerol, and initial pH levels as the most significant factors (P<0.05) for enhancing the production of chlamydospores. Subsequently, we applied the Box-Behnken design to study the interactions between, and optimal levels of, a number of factors in chlamydospore production. These statistically predicted results indicated that the highest number of chlamydospores (3.6×10⁸ spores/ml) would be obtained under the following condition: corn flour 62.86 g/L, glycerol 7.54 ml/L, pH 4.17, and 6-d incubation in liquid fermentation. We validated these predicted values via three repeated experiments using the optimal culture and achieved maximum chlamydospores of 4.5×10⁸ spores/ml, which approximately a 8-fold increase in the number of chlamydospores produced by T. harzianum SH2303 compared with that before optimization. These optimized values could help make chlamydospore production cost-efficient in the future development of novel biocontrol agents.     

Improving the antioxidant activity and enriching salvianolic acids by the fermentation of <Emphasis Type="Italic">Salvia miltiorrhizae</Emphasis> with <Emphasis Type="Italic">Geomyces luteus</Emphasis>

The antioxidant activities and total phenolic content of fermented Salvia miltiorrhiza with fungus Geomyces luteus were investigated. The results revealed that G. luteus fermentation could significantly improve the antioxidant activity and total phenolic content of S. miltiorrhiza. The main antioxidant constituents were characterized by spectroscopic analysis as salvianolic acids. High-performance liquid chromatography (HPLC) quantification also showed the enhanced content of salvianolic acid B after fermentation. The present study suggests that G. luteus fermentations are effective in the S. miltiorrhiza salvianolic acids’ enrichment process.     

Controversial opinion: evaluation of <Emphasis Type="Italic">EGR1</Emphasis> and <Emphasis Type="Italic">LAMA2</Emphasis> loci for high myopia in Chinese populations

Functional studies have suggested the important role of early growth response 1 (EGR1) and Laminin α2-chain (LAMA2) in human eye development. Genetic studies have reported a significant association of the single nucleotide polymorphism (SNP) in the LAMA2 gene with myopia. This study aimed to evaluate the association of the tagging SNPs (tSNPs) in the EGR1 and LAMA2 genes with high myopia in two independent Han Chinese populations. Four tSNPs (rs11743810 in the EGR1 gene; rs2571575, rs9321170, and rs1889891 in the LAMA2 gene) were selected, according to the HapMap database (http://hapmap.ncbi.nlm.nih.gov), and were genotyped using the ligase detection reaction (LDR) approach for 167 Han Chinese nuclear families with extremely highly myopic offspring (<?10.0 diopters) and an independent group with 485 extremely highly myopic cases (<?10.0 diopters) and 499 controls. Direct sequencing was used to confirm the LDR results in twenty randomly selected subjects. Family-based association analysis was performed using the family-based association test (FBAT) software package (Version 1.5.5). Population-based association analysis was performed using the Chi-square test. The association analysis power was estimated using online software (http://design.cs.ucla.edu). The FBAT demonstrated that all four tSNPs tested did not show association with high myopia (P>0.05). Haplotype analysis of tSNPs in the LAMA2 genes also did not show a significant association (P>0.05). Meanwhile, population-based association analysis also showed no significant association results with high myopia (P>0.05). On the basis of our family- and population-based analyses for the Han Chinese population, we did not find positive association signals of the four SNPs in the LAMA2 and EGR1 genes with high myopia.     

Antioxidant activity of alcohol aqueous extracts of <Emphasis Type="Italic">Crypthecodinium cohnii</Emphasis> and <Emphasis Type="Italic">Schizochytrium</Emphasis> sp.

Crypthecodinium cohnii (dinoflagellate) and Schizochytrium sp. (thraustochytrid) are the main sources for docosahexaenoic acid (DHA). The present study aimed to evaluate the antioxidant activity of petroleum ether, ethyl acetate, n-butanol, and water fractions of alcohol aqueous extracts of these two microalgae and to provide a theoretical basis for comprehensive utilization. The antioxidant activity was determined by total antioxidant capacity (TAC) determination, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, ferrous ion-chelating ability (FICA) assay, and reducing power (RP) assay. The total phenolic content (TPC) and total flavonoid content (TFC) were also measured by the Folin-Ciocalteu and spectrophotometry methods, respectively. The results indicated that the extracts from these two microalgae possessed good antioxidant capacity. Analysis showed that most antioxidant performance indicators (TAC, DPPH, and RP) were positively correlated with the TPC of the extracts, suggesting that the phenolics might be the major components in C. cohnii and Schizochytrium sp., contributing to their antioxidative function. Therefore, the polar fractions of C. cohnii and Schizochytrium sp. could be further examined and considered for application in health products or cosmetics.     

Antibiotic resistance mechanisms of <Emphasis Type="Italic">Myroides</Emphasis> sp.

Bacteria of the genus Myroides (Myroides spp.) are rare opportunistic pathogens. Myroides sp. infections have been reported mainly in China. Myroides sp. is highly resistant to most available antibiotics, but the resistance mechanisms are not fully elucidated. Current strain identification methods based on biochemical traits are unable to identify strains accurately at the species level. While 16S ribosomal RNA (rRNA) gene sequencing can accurately achieve this, it fails to give information on the status and mechanisms of antibiotic resistance, because the 16S rRNA sequence contains no information on resistance genes, resistance islands or enzymes. We hypothesized that obtaining the whole genome sequence of Myroides sp., using next generation sequencing methods, would help to clarify the mechanisms of pathogenesis and antibiotic resistance, and guide antibiotic selection to treat Myroides sp. infections. As Myroides sp. can survive in hospitals and the environment, there is a risk of nosocomial infections and pandemics. For better management of Myroides sp. infections, it is imperative to apply next generation sequencing technologies to clarify the antibiotic resistance mechanisms in these bacteria.     

中国地区孢子丝菌临床分离株的分子鉴定(英文)简

In this study, we investigated the molecular phylogeny of 64 clinical isolates which were identified as Sporothrix schenckii sensu lato by morphological identification. All of the strains were isolates from patients from several provinces in China. The phylogeny was inferred by DNA sequence analyses based on datasets of the ribo- somal internal transcribed spacer （ITS） and a combined ITS and partial 13-tubulin region. Reference sequences were retrieved from GenBank. Results showed that all of the isolates were clustered in a distinct clade with a type of Spo- rothrix globosa. Our analysis showed that S. globosa is the causal agent of the tested sporotrichosis in China, rather than S. schenckii that was generally believed to be the case. The existence of S. schenckii in China remains to be confirmed. This study improved our understanding of the distribution of the species in S. schenckii complex.     

Biosynthesis of (<Emphasis Type="Italic">R</Emphasis>)-2-hydroxy-3-phenylpropionic acid using whole recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> cells in an aqueous/<Emphasis Type="Italic">n</Emphasis>-octane biphasic system

(R)-2-hydroxy-3-phenylpropionic acid (PLA) is an ideal antimicrobial compound with broad-spectrum activity against a wide range of Gram-positive bacteria, some Gram-negative bacteria, and fungi. We studied the bioconversion of phenylpyruvate (PPA) to PLA using whole recombinant Escherichia coli cells in a series of buffer/organic solvent systems. Octane was found to be the best organic solvent. The optimum volume ratio of the water phase to the n-octane phase, conversion temperature, substrate concentration, and cell concentration were 6:4, 40 °C, 12.5 g/L, and 30 g/L wet cells, respectively. Under the optimized conditions, the average PLA productivity in the aqueous/n-octane system was 30.69% higher than that in the aqueous system, and 32.31 g/L PLA was obtained with the use of a stirred reactor (2-L scale). Taken together, our findings indicated that PLA biosynthesis was more efficient in an aqueous/n-octane biphasic system than in a monophasic aqueous system. The proposed biphasic system is an effective strategy for enhancing PLA yield and the biosynthesis of its analogues.     

On the Relationship Between <Emphasis Type="Italic">Belief</Emphasis> and <Emphasis Type="Italic">Acceptance</Emphasis> of Evolution as Goals of Evolution Education

The issue of the proper goals of science education and science teacher education have been a focus of the science education and philosophy of science communities in recent years. More particularly, the issue of whether belief/acceptance of evolution and/or understanding are the appropriate goals for evolution educators and the issue of the precise nature of the distinctions among the terms knowledge, understanding, belief, and acceptance have received increasing attention in the 12 years since we first published our views on these subjects. During that time, our own views about these issues have evolved, and this article presents a reconsideration of both these distinctions and the propriety of these goals. In particular, the present paper continues our discussion of the nature of belief as it relates to science education, and more specifically to evolution education. We extend that work to consider the import of the distinction between belief and acceptance.     

Polymorphisms of the <Emphasis Type="Italic">IGF1</Emphasis> gene and their association with growth traits,serum concentration and expression rate of <Emphasis Type="Italic">IGF1</Emphasis> and <Emphasis Type="Italic">IGF1R</Emphasis> in buffalo

The insulin-like growth factor 1 (IGF1) gene is a member of the group of somatotropin axis genes that play a significant role in cell proliferation and growth of muscles. Here, we searched for polymorphisms in buffalo IGF1 and found two novel single nucleotide polymorphisms (SNPs), G64A and G280A, in the noncoding sequences of exon 1 and exon 4, respectively. Statistical analysis of different genotypes showed that the individuals with GG genotypes had significantly (P<0.05) higher body weight (BW) and average daily gain (ADG) than those with other genotypes at ages of 3–6 months in G64A SNP and 6–9 months in G280A SNP. The combined genotypes of these two SNPs produced three haplotypes, GG/GG, AG/AG, and AA/AA, which were significantly associated (P<0.0001) with BW and ADG at an age from 3 to 12 months. Buffaloes with the homozygous GG/GG haplotype showed higher growth performance than other buffaloes. The two SNPs were correlated with mRNA levels of IGF1 and IGF1 receptor (IGF1R) in semitendinosus muscle as well as with the serum concentration level of IGF1. Also, buffaloes with GG/GG haplotype showed higher mRNA and serum concentration levels. The data revealed that these two SNPs could be valuable genetic markers for selection of Egyptian buffaloes for better performance in the population.     

Antimelanogenic effects of <Emphasis Type="Italic">Inula britannica</Emphasis> flower petal extract fermented by <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> KCCM 11613P

The inhibitory effects of Lactobacillus plantarum-fermented and non-fermented Inula britannica extracts on the tyrosinase activity were comparatively investigated to examine whether and how they improve the whitening activity, and the contents of total flavonoids and polyphenolics as bioactive compounds were determined. The skin whitening activity using in vitro or ex vivo tyrosinase and L-3,4-dihydroxyphenylalanine (L-DOPA) staining was examined. The total flavonoid content (TFC) was increased by 13.4% after 72 h-fermentation. The viabilities of the B16F10 cells treated with the fermented and non-fermented control extracts were 100.26% and 92.15% at 500 μg/ml, respectively. In addition, the inhibition of tyrosinase activity was increased by the fermented samples from 29.33% to 41.74% following fermentation for up to 72 h. The tyrosinase activity of the untreated control group was increased to 145.69% in B16F10 cells. The results showed that I. britannica fermented by L. plantarum dose-dependently inhibited tyrosinase activity, which was stimulated by α-melanocyte stimulating hormone. These results suggest that lactic fermented I. britannica extracts can be used as effective skin-whitening materials.     

Antibacterial and anti-biofilm activity of the lipid extract from <Emphasis Type="Italic">Mantidis ootheca</Emphasis> on <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis>

The aim of this study is to assess the antibacterial and anti-biofilm properties of the lipid extract from Mantidis ootheca against the gentamycin resistant Pseudomonas aeruginosa. The chemical composition of the lipid extract and its relative proportion were determined using the technique of gas chromatography coupled with mass spectrometry (GC-MS). Antibacterial susceptibility tests were performed using a disc diffusion assay and the minimum inhibition concentration (MIC) was determined by way of the agar dilution method. The anti-biofilm test was carried out with crystal violet staining and scanning electron microscopy (SEM). There were 16 compounds detected, and the most abundant components were sesquiterpenoids, monoterpenes, and trace aromatic compounds. The MIC for P. aeruginosa was 4 mg/ml and the eradication effect on preformed biofilms was established and compared with a ciprofloxacin control. The results of our study indicated that a lipid extract from M. ootheca could be used as a topical and antibacterial agent with anti-biofilm activity in the future.     

<Emphasis Type="Italic">Pediococcus Acidilactici</Emphasis> Inhibit Biofilm Formation of Food-Borne Pathogens on Abiotic Surfaces

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 4 log 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.     

Non-dispersive solvent extraction of <Emphasis Type="Italic">p</Emphasis>-toluic acid from purified terephthalic acid plant wastewater with <Emphasis Type="Italic">p</Emphasis>-xylene as extractant

Non-dispersive solvent extraction (NDSE) with p-xylene as extractant was employed as a novel separation method to recover both p-toluic (PT) acid and water from purified terephthalic acid (PTA) wastewater. The mass transport behavior of PT acid from aqueous solution to p-xylene was investigated by experiments and numerical simulation. Experiments showed that NDSE is feasible and effective. Residual PT acid in the raffinate can be reduced to lower than the permitted limit of wastewater re-use (100 g/m³) with extraction time longer than 60 s in industrial conditions. A mathematical model of PT acid mass transport was developed to optimize the membrane module performance. The model was validated with the experimental results with relative errors of less than 6%. Numerical analysis for mass transfer through the lumen side, the porous membrane layer, and the shell side showed that PT acid transport in the aqueous solution is the rate determining step. The effects of the membrane and operating parameters on membrane module performance were investigated by means of computational simulations. The key parameters suggested for industrial NDSE design are: fiber inner radius r₁=200–250 µm, extraction time t_e=50–60 s, aqueous/organic volumetric ratio a/o=9.0, and temperature T=318 K.     

Meta-analysis of <Emphasis Type="Italic">C242T</Emphasis> polymorphism in <Emphasis Type="Italic">CYBA</Emphasis> genes: risk of acute coronary syndrome is lower in Asians but not in Caucasians

Background

A lot of studies have demonstrated that C242T polymorphism in CYBA genes may play an important role in the pathological process of acute coronary syndrome (ACS). However, the results are not consistent. To further evaluate this debate, we performed a meta-analysis to determine the relationship between C242T polymorphism and ACS.

Methods and results

We screened PubMed/MEDLINE, EBSCIO, and EMBASE research reports until Mar. 2014 and extracted data from 10 studies involving 6102 ACS patients and 8669 controls. Subgroup analysis by ethnicity documented a significant decreased risk of ACS for C242T polymorphism in the Asian population under allelic comparison (odd ratio (OR) 0.73; 95% confidence intervals (CI) 0.64–0.83), dominant model (OR 0.71; 95% CI 0.62–0.82), and homozygote comparison (OR 0.57; 95% CI 0.35–0.92). However, in the overall population and especially with Caucasians, no significant association was uncovered. Further meta-regression analysis revealed that the heterogeneity among studies was largely attributed to ethnicity. No publication bias was detected through a funnel plot and an Egger’s linear regression test.

Conclusions

Taken together, our results suggest that the C242T polymorphism might be a protective factor against developing ACS in the Asian population. Further researches will be needed to identify the confounding factors which modified the protective effect of T allele among Caucasians.