Determination of Whole Molecular of Thermostable Direct Hemolysins in Milk Powder by HPLC-ESI-TOF.

Although Vibrio parahaemolyticus (V. parahaemolyticus) is a pathogen frequently found in seafood, there is a possibility of its presence in other foods, such as dairy products. The main virulence factors of V. parahaemolyticus are thermostable direct hemolysins (TDHs) which are lethal toxins, so it is necessary to establish qualitative and quantitative methods for determining TDHs. HPLC-ESI-TOF was employed to establish a method for identifying TDHs. The identification and quantification ions of TDHs were confirmed by HPLC-ESI-TOF. The method was developed for detecting TDHs in milk powder using HPLC-ESI-TOF in this paper, and limits of detection (were between 0.20 and 0.40 mg/kg, limits of quantitation were between 0.5 and 1.0 mg/kg and recoveries of all TDHs were between from 78% to 94% with relative standard deviation lower than 10%. This research will provide a reference for developing methods of HPLC-MS/MS to detect TDHs in food samples, which can provide a tool for the government to monitor TDHs contamination in foods.

Combatting cyanobacteria: unraveling the potency of 316L-Cu stainless steel in inhibiting Microcystis aeruginosa growth.

Harmful algal blooms, particularly those of Microcystis aeruginosa, present significant ecological and health risks. To address this issue, this study utilized a custom static algal growth assessment apparatus to investigate the anti-algal performance of a copper-alloyed 316L stainless steel (SS), named 316L-Cu SS. This material was compared with traditional 316L SS, which is widely utilized in freshwater systems for its corrosion resistance. Algal growth dynamics were monitored through optical density (OD) and chlorophyll A concentration measurements. Notably, 316L-Cu SS exhibited superior inhibitory effects on Microcystis aeruginosa growth compared to 316L SS and control groups. Inductively coupled plasma mass spectrometry (ICP-MS) confirmed that the copper ion release from 316L-Cu SS played a critical role in this algal suppression, which interfered with photosynthesis, induced oxidative stress, and damaged algal cell membranes. In contrast, other metal ions (Ni, Cr, Fe) had a negligible impact on algal growth. The study highlights 316L-Cu SS as a promising material for mitigating harmful algal blooms, thereby offering potential benefits for both aquatic ecosystem conservation and public health protection.

Effect of environmental factors on expression of staphylococcal enterotoxin genes.

Staphylococcal enterotoxins (SEs) secreted by Staphylococcus aureus (S. aureus) can cause foodborne disease, nausea, vomiting and diarrhea, and even death. Regulation of SE expression is related to accessory gene regulators (Agr). It is important to reveal which environmental factors influence regulation of SE expression to prevent SE food poisoning outbreak. Hence, natural environmental factors which may have an impact on SE expression were selected, such as temperature, food types, strains, and competing strains. Seven strains of S. aureus carrying different SE genes were collected from the Chinese Academy of Inspection and Quarantine (CAIQ) strain bank for study. Strains were cultured with different conditions. Temperature was 8 °C, 22 °C, and 30 °C. Food type was milk powder and nutrient broth. Competing strains were Vibrio parahaemolyticus (V. parahaemolyticus), Escherichia coli (E. coli), and Bacillus cereus (B. cereus). The expression culture solution was pretreated by centrifugation, then determined by using SDS-PAGE, and distinguished SEs apart from each other by HPLC-ESI-TOF. There are 168 samples collected from SE expression culture; the result of SDS-PAGE suggests 23 samples were positive for SEs, and the other 145 samples were negative for SEs. The result of HPLC-ESI-TOF suggests that SEs with similar molecular weight can be distinguished in terms of m/z. The most important factor contributing to regulate expression of SEs was estimated by logistic regressive analysis. The result shows that McFadden R2 is 0.213; p value is 0.000 (p < 0.05); this result illustrates that the model is valid and meaningful. Strains, food types, temperature, and competing strands can explain the 21% change in SE expression. Temperature (z = 3.029, p = 0.002 < 0.01), strains (z = - 3.132, p = 0.002 < 0.01), and food types (z = - 2.415, p = 0.016 < 0.05) have significant impact on SE expression, and the competing strains (z = 1.230, p = 0.219 > 0.05) have no impact on the SE expression. More important impact on SE expression was estimated by OR value; the result shows that strength of temperature influencing on SE expression is bigger than strains and food types in terms of values of OR, temperature (OR = 2.862), strains (OR = 0.641), and food types (OR = 0.561); consequently, temperature is a key factor for stimulating SE expression and had high expression at 30 °C. Therefore, food easily contaminated with S. aureus should be monitored intensively at early and late summer, when proper temperature for expressing SEs may result in S. aureus food poisoning prevalence.

A Super Uniform Hydrophobic Gas Diffusion Layer for a Proton Exchange Membrane Fuel Cell.

The design and optimization of the gas diffusion layer (GDL) play a crucial role in the improvement of proton exchange membrane fuel cell performance. Hydrophobic treatment of a GDL is an important method for facilitating mass transfer, while conventional Teflon treatment is not uniform and leads to an increase in ohmic and heat resistance. Herein, a homogeneous molecular hydrophobic GDL was prepared by liquid phase synthesis, and a two-dimensional non-isothermal model was developed to investigate the transfer mechanism. The peak power density of cells with the GDL described above was improved by 46% compared to that of the conventional GDL. The ohmic and mass transport resistance decreased by 15% and 52%, respectively, under a current density of 1 A cm-2 using the uniform hydrophobic GDL. The simulation results proved that the uniform hydrophobic GDL eliminates the hydrophilic dots, which prevents the formation of water pools and reduces the resistance to gas flow. The water saturation of the conventional GDL reaches 0.19 at a current density of 1 A cm-2, and the saturation of a modified GDL under the same conditions is only 0.13. A dimensionless parameter, Tf, is proposed to characterize the resistance of oxygen diffusion. In conclusion, molecular-level uniform hydrophobic treatment can effectively reduce the ohmic and mass transfer resistance of a GDL and effectively improve the performance of fuel cells.

Structure and Wear Performance of a Titanium Alloy by Using Low-Temperature Plasma Oxy-Nitriding.

To solve the problems of high nitriding temperature and long nitriding time with conventional plasma nitriding technologies, a kind of low-temperature plasma oxy-nitriding technology containing two-stage processes with different ratios of N to O was developed on a TC4 alloy in this paper. A thicker permeation coating can be obtained with this new technology compared to conventional plasma nitriding technology. The reason for this is that the oxygen introduction in the first two-hour oxy-nitriding step can break the continuous TiN layer, which facilitates the quick and deep diffusion of the solution-strengthening elements of O and N into the titanium alloy. Moreover, an inter-connected porous structure was formed under a compact compound layer, which acts as a buffer layer to absorb the external wear force. Therefore, the resultant coating showed the lowest COF values during the initial wear state, and almost no debris and cracks were detected after the wear test. For the treated samples with low hardness and no porous structure, fatigue cracks can easily form on the surface, and bulk peeling-offcan occur during the wear course.

Investigation on Potential Biomarkers and Immune-Related Mechanisms of Ischemic Stroke.

BACKGROUND: This study aimed to reveal the detailed immune-related mechanisms underlying ischemic stroke (IS) and identify new immune-associated biomarkers for clinical management. METHODS: Differentially expressed genes (DEGs) between IS samples and normal controls were identified using the GSE16561 dataset. The feature genes of the immune cells were investigated using the GSE72642 dataset. Weighted correlation network analysis (WGCNA) was performed to reveal module genes, followed by an investigation of common DEGs and a functional enrichment analysis. Potential biomarkers were identified based on hub genes in protein-protein interaction networks and WGCNA. Finally, GSE158312 was used for biomarker verification. RESULTS: In total, 1230 DEGs were identified between the IS samples and normal controls. Seven clinically significant modules were identified using WGCNA. The yellow module genes were positively correlated with polymorphonuclear cells (PMNC), whereas the brown module genes were positively correlated with CD4+ T cells. Eight genes were selected as hub genes. These genes are mainly involved in functions such as the innate immune response. Upregulated TLR2 and ARG1 levels were significantly different between the two groups in the verification dataset. CONCLUSIONS: Our findings suggest ARG1 and TLR2 as novel biomarkers for IS. Upregulated TLR2 might play a role in IS development by participating in the innate immune response function.

Novel Fe-Based Amorphous Composite Coating with a Unique Interfacial Layer Improving Thermal Barrier Application.

To improve thermal barrier applications in advanced vehicle engines, a novel Fe-based amorphous composite coating was designed by introducing ceramic oxides and was prepared by atmospheric plasma spraying (APS). The microstructure and related properties of the as-deposited coating were investigated in detail. The composite coating comprises a well-formed FeCrNbBSi amorphous metallic matrix and dispersed yttria-stabilized zirconia (YSZ) splats. A unique Si-oxide interfacial layer with a thickness of several nanometers and an amorphous structure forms between the metallic matrix and ceramic phase, which is attributed to a combination of multiple effects. The composite coating displays extremely low thermal conductivity from 2.28 W/mK at 100 °C to 3.36 W/mK at 600 °C and can increase the surface temperature of the piston crown by 18.93 °C, which implies a significant means of enhancing the power efficiency. The improved thermal barrier ability of the composite coating is revealed as the crucial effect of the Si-oxide interfacial layer, which induces an increased interfacial thermal resistance. The fracture toughness of the composite coating remains at 3.40 MPa·m1/2, comparable to that of the monolithic amorphous coating, 3.74 MPa·m1/2, which is closely related to the formation of a Si-oxide layer and its nanoscale thickness. Therefore, the Fe-based amorphous composite coating developed here demonstrates great potential as an innovative metal-based thermal barrier coating for application in vehicle engines and provides specific inspiration for future works exploring the interfacial engineering of coating.

Advanced Reversal Tolerant Anode in Proton Exchange Membrane Fuel Cells: Study on the Attenuation Mechanism during Fuel Starvation.

Fuel starvation at the anode of a proton exchange membrane fuel cell can lead to the increase of anode potential and the reversal of cell voltage followed by water electrolysis and carbon corrosion. A material-based approach (with high active water electrolysis catalysts) does not have much influence on the electrochemical performance, and carbon corrosion can be effectively avoided compared with the complex active control system. However, the membrane electrode assembly shows poor reversal tolerant performance during the hydrogen starvation test in previous studies, and the degradation mechanism is unclear. Therefore, reversal tolerant anode electrodes are designed in this article, and the voltage decrease mechanism is investigated comprehensively. The results exhibit that the increase of anode potential is mainly caused by the increase of mass transport resistance. Additionally, the voltage reversal time can be up to 5020 min, and the degradation rate of cell voltage at 1.2 A cm-2 can be as low as 0.12% h-1 after the first fuel starvation test.

Development and verification of a quantitative real-time PCR method to identify and quantify gelatin derived from animal hide.

The species origin of hide gelatin is a crucial issue with respect to health concerns and religious restrictions. Analysis of the animal-derived ingredients of gelatin by reliable methods is necessary to ensure its authenticity. However, due to the highly processed nature of gelatin, it remains a challenge to identify gelatin end products accurately and robustly. Our study established and verified a quantitative real-time PCR (qPCR) method based on careful selection of target genes and a DNA extraction method. The middle products of the gelatin production streamline were investigated to explore the influence of each critical processing step on the method. Gelatin reference samples were used to quantify the levels of target species. Commercial gelatin commodities were surveyed to highlight the mislabeling situation. In summary, the qPCR method was demonstrated to be highly specific and sensitive, with limits of detection (LOD) of 0.1 to 1 pg/µL and gelatin LODs of 0.1% to 5% (w/w). The transition from decoction to concentrated gel was found to have the most severe effect on the qPCR. Intensification of pressure or temperature or employment of enzyme hydrolysis aggravated the DNA damage, resulting in elevated Cq values. Quantitation of gelatin products was feasible; gelatin products produced from 5% target hide and 95% matrix hide mixtures showed 2.9% to 5.2% target species. The 26% relative error for low gelatin content is acceptable for semiquantitation purposes. A market survey showed that 52.6% of the gelatin products were mislabeled as being of animal origin.

Chemical and cytological evaluation of honeybee pollen antioxidant ability.

The crude flavonoid extract of pollen (CFP) of four species of honeybee pollens were extracted with ethanol, and the total flavonoid contents ranged from 3.4 to 14.5 mg rutin/g dry weight. The antioxidant activities of the CFPs were evaluated from both chemical and cytological aspects. Comprehensive antioxidant scores were determined based on these two evaluation systems. The results showed that canola CFP had the highest antioxidant capacity among the four CFPs. A cytotoxicity assay was conducted to assess the safety threshold of the CFPs, and canola CFP was proved to be the least toxic to vascular endothelial cell. Of the four tested CFPs, this research suggests that canola CFP is the most promising natural antioxidant. In addition, high-performance liquid chromatography (HPLC) analysis detected seven flavonoid glycosides in the hydrolysates of the four CFPs. Among them, quercetin and kaempferol were present in all four honeybee pollen extracts, but there were significant differences between their contents. A correlation analysis revealed a strong correlation between the content of quercetin in the pollen extract and the extract's antioxidant activity. PRACTICAL APPLICATION: Many varieties of honeybee pollen are commercially available. The results of this study help guide consumers to choose honeybee pollens that have a better antioxidant effect. This report can also provide guidance and data in support of the development of honeybee pollen health products.

Three-Dimensional Superlithiophilic Interphase for Dendrite-Free Lithium Metal Anodes.

Lithium metal is among the most promising anode candidates of high-energy-density batteries. However, the formed dendrites result in low Coulombic efficiency and serious security issues. Designing lithiophilic sites is one of the effective strategies to control Li deposition. Herein, we propose a three-dimensional lithiophilic N-rich carbon nanofiber with the decoration of ZnO granules as a protective layer for a dendrite-free lithium metal anode. Theoretical evaluation indicates the synergistic effects of lithiophilic ZnO and N-containing functional groups enhance lithium adsorption and trigger uniform deposition. With the lithiophilic interlayer, the lithium deposition overpotential is only ∼20, 50, and 74 mV at 1, 3, and 6 mA cm-2, respectively, which are much lower than those without the functional interlayer (∼55, 130, and 238 mV). The average Coulombic efficiency of lithium stripping and plating is up to ∼97.4% (94.0% for that without the interlayer) at 0.5 mA cm-2. Meanwhile, the Li|LiFePO4 full cell with the superlithiophilic interlayer demonstrates a high capacity retention rate of 99.6% (91.0% for that without the interlayer) over 200 cycles at 1 C. The introduction of the lithiophilic interphase could provide a convenient strategy and guidance to design the configuration for the practical application of Li metal batteries.

Fluorophosphorus derivative forms a beneficial film on both electrodes of high voltage lithium-ion batteries.

Layered lithium-rich oxides, as a series of highly promising cathode material for lithium-ion batteries, attract extensive attention due to their high specific capacity and high working potential (4.6 V vs Li/Li+). However, the poor interface stability of the cathode and electrolyte seriously restricts their practical application. In this article, theoretical calculations, linear sweep voltammetry and cyclic voltammetry results indicate that tris (pentafluorophenyl) phosphine (TPFPP) is a potential dual-functional electrolyte additive to solve interface problems. The TPFPP additive can decompose preferentially on the surface of both electrodes and form uniform and stable protective films, which effectively inhibit the continuous decomposition of the electrolyte and significantly alleviate the dissolution of transition metal ions during cycling. Owing to the above effects, the capacity retention and coulombic efficiency of Li1.17Ni0.25Mn0.58O2 (LLO)/graphite (Gr) cells are improved from 62.6% and 97.7% to 90.6% and 99.8% after 200 cycles at 0.3 C (1 C = 300 mA g-1), respectively. This study provides a wide prospect for the application of lithium-rich materials in the future.

Applying COI Barcode to Identify Animal Origin of Food.

DNA barcoding possesses advantages of high resolution, high sensitivity, and capability in capturing as much identity information as possible. However, highly varying sources of food materials and a complicated supply chain bring about challenge to the application of barcoding methods. In this study, different barcode systems were compared to establish a robust method for tracing animal species in food. Experiments on food samples from mammal, poultry, and fish proved that a mini barcode system targeting a 192 bp COI gene fragment was able to accurately identify both raw and highly processed animal food. In order to distinguish species in a mixed food sample, cloning technique was used by which as low as 10% target animal ingredient could be detected. Testing of marketed food products verified the capability of the mini barcoding method in identifying illegally claimed product.

Identification of Small Berry Species in Food and Juice Using TaqMan-Based Real-Time PCR.

Background: Public interest is growing for small berries in recent years because they are very delicious, low in energy, and full of bioactive compounds with potential health benefits. Similar to other food products, adulteration of small berry fruit products poses economic and safety problems to consumers. Objective: To protect consumers and regulate the small berry fruit products market, it is necessary to establish a robust method for detecting the authenticity. Methods: In this study, TaqMan-based real-time PCR assay was established for species identification of cranberry, raspberry, and blueberry to ensure authenticity of commercial small berry food products (pulp, dried fruit, fruit juice, jam, and puree). Results: Absolute detection limit was 0.1 pg/µL DNA for raspberries, 1 pg/µL DNA for blueberries, and 10 pg/µL DNA for cranberries. Practical LOD was 0.1% (v/v) for fresh juice. For processed juice, practical LOD was 1% for blueberry and red raspberries, 0.1% for black and yellow raspberries, and 5% for cranberry. Conclusions: The method was shown to be functional and effective to detect the raw material composition of cranberry, raspberry, and blueberry for commercial products. Highlights: TaqMan probe-based real-time PCR methods were designed to identify three small berries (blueberry, raspberry, and cranberry) in berry products. Efficient DNA recovery methods and detection strategy were established to ensure correct and sensitive testing of fresh small berries exhibited a detection limit of 0.1 to 10 pg/µL. The practical minimum detection levels were 0.1 to 5% in fresh and processed juice, including pasteurization and HTHP.

Authentication of Small Berry Fruit in Fruit Products by DNA Barcoding Method.

Small berry fruit products are gaining an expanded market due to their high nutrition value. However, the authenticity of products is challenged by adulteration and mislabeling. To establish an accurate and robust method for identifying both known and unknown fruit species in small berry fruit products, DNA barcoding technology based on Sanger sequencing was adopted. To overcome the influence of processing conditions on DNA recovery, mini-barcodes of rbcL and ITS and a medium-barcode of psbA-trnH were applied. To identify ingredients in products containing mixed species, plasmid cloning was applied to separate mixed barcodes. The method established in this paper could detect 1% to 10% target species in mixed fruit juice.

A subchronic feeding safety evaluation of transgenic milk containing human ß-defensin 3 on reproductive system of C57BL/6J mouse.

Bovine mastitis is an infectious disease of the mammary gland which has been generally treated by antibiotic delivery. While the increasing drug-resistant bacteria and the high consumption of the antibiotic had become a noticeable concern. In a previous study, a mammary special vector expressing human ß-defensin 3 (hBD3) was transfected into bovine fetal fibroblasts to produce mastitis-resistant bovine. This investigation focused on potential unintended effects of transgenic milk containing hBD3 produced by these mastitis-resistant bovine on the reproductive system of C57BL/6J mice. Mice were fed with diets containing transgenic milk or conventional milk, nutritionally balanced to an AIN93G diet for 90 days, and non-milk diet was selected as the negative group. The reproductive system was given special attention including reproductive organ/body ratios, necropsy and histopathology, serum sex hormone, sperm parameters, estrus cycle and the expression level of some specific genes which could indicate the development and function of reproductive system. No diet-related significant differences were observed among three groups in this 90-day feeding study. The results indicated that hBD3 milk does not appear to exert any effect on the reproductive system in C57BL/6J rats compared with conventional milk or the control diet.

Toward a Slow-Release Borate Inhibitor To Control Mild Steel Corrosion in Simulated Recirculating Water.

On the basis of their potential passivating characteristics, in this study, borates have been used to synthesize a novel slow-release inhibitor to suppress long-term mild steel corrosion in simulated recirculating water. The passivating performance was characterized by various electrochemical measurements and the passivating mechanism was interpreted by the point defect model. The experimental results indicated that the slow-release inhibitor exhibited a passivating efficiency of over 98% after 30 days of immersion, due to the formation of a passive film that was predominant by a Fe-O-B structure on the mild steel surface. This study provides a novel controlled-release concept and a slow-release borate inhibitor to control long-term corrosion.

Molecular Tracing of the Origin of Six Different Plant Species in Bee Honey Using Real-Time PCR.

The quality of honey is significantly influenced by floral origin. Mislabeling floral species occurs frequently in bee honey products. To protect consumers from economic fraud and maintain a fair market environment, methods to identify floral species in honey are necessary. In our study, real-time PCRs were established, targeting six honey types mainly produced in China (canola, Chinese milkvetch, Chinese chaste tree, locust tree, litchi, and longan). Sensitivity testing on DNA from plant tissues exhibited LODs of about 0.5-5 pg/µL. For DNA extracts of pollen sediments from different honey species, LODs ranged from 13.6 to 403.2 pg/µL. In an experiment to determine the practical LODs of honey in which adulterant honey was spiked in the genuine honey, adulterant honey as low as about 0.1-0.5% was detected in 90-100% in 10 parallel tests. Additionally, pollen was spiked in the honey and stored under various conditions to investigate the migration of pollen DNA into the honey supernatant. Finally, the efficiency of our method was investigated by testing honey samples of unknown compositions from different geographic regions. Of the 159 honey samples that were supposed to be monofloral that had been collected in five provinces, a small portion were found to be contaminated with foreign pollen (7%). The methods proved to be specific, sensitive, and reliable in identifying the six plant species in honey, which would be a useful tool during the market supervision and QC of honey products.

Effects of Genetically Modified Milk Containing Human Beta-Defensin-3 on Gastrointestinal Health of Mice.

This study was performed to investigate the effects of genetically modified (GM) milk containing human beta-defensin-3 (HBD3) on mice by a 90-day feeding study. The examined parameters included the digestibility of GM milk, general physical examination, gastric emptying function, intestinal permeability, intestinal microflora composition of mice, and the possibility of horizontal gene transfer (HGT). The emphasis was placed on the effects on gastrointestinal (GI) tract due to the fact that GI tract was the first site contacting with food and played crucial roles in metabolic reactions, nutrition absorption and immunity regulation in the host. However, the traditional methods for analyzing the potential toxicological risk of GM product pay little attention on GI health. In this study, the results showed GM milk was easy to be digested in simulated gastric fluid, and it did not have adverse effects on general and GI health compared to conventional milk. And there is little possibility of HGT. This study may enrich the safety assessment of GM product on GI health.

A Real-Time PCR Method Targeting Camel Ingredient for Food Authentication.

The special nutritious value of camel showed high potential for market exploitation. In this paper, a real-time PCR method targeting camel ingredient in camel meat and milk is reported as an approach to fight against adulteration. To understand the impact of processing procedures on the amplifiability of cytb gene, four kinds of processed camel meat were investigated, and the rate of DNA breakage was explored. The method was able to detect 5 fg/µL camel DNA and highly processed food containing 0.01% camel meat with a high confidence level.