Methionine Promotes Milk Protein Synthesis via the PI3K-mTOR Signaling Pathway in Human Mammary Epithelial Cells.

Breast milk is widely considered to be the most natural, safe, and complete food for infants. However, current breastfeeding rates fall short of the recommendations established by the World Health Organization. Despite this, there are few studies that have focused on the promotion of human lactation through nutrient supplementation. Therefore, the aim of this study was to investigate the effect of methionine on milk synthesis in human mammary epithelial cells (MCF-10A cells) and to explore the underlying mechanisms. To achieve this, MCF-10A cells were cultured with varying concentrations of methionine, ranging from 0 to 1.2 mM. Our results indicated that 0.6 mM of methionine significantly promoted the synthesis of milk protein. An RNA-seq analysis revealed that methionine acted through the PI3K pathway. This finding was validated through real-time quantitative polymerase chain reaction (RT-qPCR) and Western blotting. In addition, PI3K inhibition assays confirmed that methionine upregulated the expression of both mTOR and p-mTOR through activation of PI3K. Taken together, these findings suggest that methionine positively regulates milk protein synthesis in MCF-10A cells through the PI3K-mTOR signaling pathway.

Zeolitic Imidazolate Framework-8 Composite-Based Enzyme-Linked Aptamer Assay for the Sensitive Detection of Deoxynivalenol.

The mycotoxin deoxynivalenol (DON) is a prevalent contaminant in cereals that threatens the health of both humans and animals and causes economic losses due to crop contamination. The rapid and sensitive detection of DON is essential for food safety. Herein, a colorimetric biosensor based on horseradish peroxidase- and gold nanoparticle-encapsulated zeolitic imidazolate framework-8 (HRP&Au@ZIF-8) was developed for the sensitive screening of DON. The synthesized HRP&Au@ZIF-8 probes not only held great potential for signal amplification but also exhibited stable catalytic activity even under extreme conditions, which endowed the biosensor with both good sensitivity and stability. Under the optimized conditions, qualitative measurement of DON can be achieved through visual inspection, and quantitative evaluation can be performed via absorbance measurements at a characteristic wavelength of 450 nm. The proposed method has demonstrated high sensitivity with a linear detection range of 1-200 ng/mL and a detection limit of 0.5068 ng/mL. It also presented good selectivity and reliability. Furthermore, DON in spiked cereal samples has been quantified successfully using this method. This novel approach demonstrates significant potential for the facile and expeditious detection of DON in cereal products and brings us one step closer to enhancing food safety.

The neurobehavioral impacts of typical antibiotics toward zebrafish larvae.

Due to the widely usage in livestock, aquaculture and clinics, antibiotic residues are existed in aqueous environments and their potential toxicity to aquatic organisms is concerning. Here, we used zebrafish as the model to investigate the neurotoxicity and involved mechanism of seven antibiotics that were frequently detected in surface waters. The results revealed that the short-term exposure to clarithromycin (CLA), chlortetracycline (CTC) and roxithromycin (ROX) induced behavioral effects, with effective concentration of 1 µg/L (CTC and ROX) and 100 µg/L (CLA, CTC and ROX) respectively. A significant decrease in the travel distance and velocity as well as an increase in turn angle was measured. TUNEL assay identified increased cell apoptosis in brain sections of larvae exposed to three neurotoxic antibiotics, which raised the possibility that the behavioral symptoms were associated with neural damage. Transcriptome sequencing showed that the three antibiotics could affect the nervous system of zebrafish including the alteration of synaptogenesis and neurotransmission. Additionally, ROX and CTC affected pathways involved in mitochondrial stress response and endocrine system in zebrafish larvae. Besides, BDNF, ASCL1, and CREBBP are potential upstream regulatory factors that mediated these impacts. These findings indicated that exposure of CTC, ROX and CLA may cause abnormal behavior toward zebrafish larvae under environmental relevant concentration and revealed the potential role of neural cell apoptosis and synaptogenesis signaling in mediating this effect.

Surface engineering of magnetic peroxidase mimic using bacteriophage for high-sensitivity/specificity colorimetric determination of Staphylococcus aureus in food.

Herein, we proposed surface engineering of magnetic peroxidase mimic using bacteriophage by electrostatic interaction to prepare bacteriophage SapYZU15 modified Fe3O4 (SapYZU15@Fe3O4) for colorimetric determination of S. aureus in food. SapYZU15@Fe3O4 exhibits peroxidase-like activity, catalyzing 3,3',5,5'-tetramethylbenzidine (TMB) chromogenic reaction. After introducing S. aureus, peroxidase-like activity of SapYZU15@Fe3O4 was specifically inhibited, resulting in deceleration of TMB chromogenic reaction. This phenomenon benefits from the presence of unique tail protein gene in the bacteriophage SapYZU15 genome, leading to a specific biological interaction between S. aureus and SapYZU15. On basis of this principle, SapYZU15@Fe3O4 can be employed for colorimetric determination of S. aureus with a limiting detection (LOD), calculated as low as 1.2 × 102 CFU mL-1. With this proposed method, colorimetric detection of S. aureus in food was successfully achieved. This portends that surface engineering of nanozymes using bacteriophage has great potential in the field of colorimetric detection of pathogenic bacterium in food.

Versatile dodecyl trimethyl ammonium bromide modified γ-FeOOH for simultaneous removal and determination of As(⠤).

Inorganic arsenic pollution in water spreads all over the world, tremendously threatening environmental safety and human health. Herein, versatile dodecyl trimethyl ammonium bromide modified γ-FeOOH (DTAB-γ-FeOOH) was prepared for sportive removal and visual determination of As(Ⅴ) in water. DTAB-γ-FeOOH displays a nanosheet-like structure with a high specific surface area calculated as 166.88 m2 g-1. Additionally, DTAB-γ-FeOOH shows peroxidase-mimicking feature, which can catalyze colorless TMB to generate blue oxidized TMB (TMBox) in presence of H2O2. Removal experiments show that DTAB-γ-FeOOH exhibits good As(Ⅴ) removal efficiency because modification of DTAB makes γ-FeOOH carry abundant positive charges, improving affinity between DTAB-γ-FeOOH and As(Ⅴ). It is found that theoretical maximum adsorption capacity is up to 126.91 mg g-1. Moreover, DTAB-γ-FeOOH can resist interference of most of co-existing ions. After that, As(Ⅴ) was detected based on peroxidase-like DTAB-γ-FeOOH. As(Ⅴ) can be adsorbed onto DTAB-γ-FeOOH surface, markedly inhibiting its peroxidase-like activity. Based on it, As(Ⅴ) ranging from 1.67 to 3333.33 µg L-1 can be well detected, with a low LOD (0.84 µg L-1). The successful sorptive removal and visual determination of As(Ⅴ) from real environmental water indicated that DTAB-γ-FeOOH has great potential in the treatment of As(Ⅴ)-containing environment water.

Rapid and effective detection of Macrobrachium rosenbergii nodavirus using a combination of nucleic acid sequence-based amplification test and immunochromatographic strip.

Nucleic acid sequence-based amplification (NASBA) provides a fast and convenient approach for nucleic acid amplification under isothermal conditions, and its combination with an immunoassay-based lateral flow dipstick (LFD) could produce a higher detection efficiency for M. rosenbergii nodavirus isolated from China (MrNV-chin). In this study, two specific primers and a labelled probe of the capsid protein gene of MrNV-chin were constructed. The process of this assay mainly included a single-step amplification at a temperature of 41 â„ƒ for 90 min, and hybridization with an FITC-labeled probe for 5 min, with the hybridization been required for visual identification during LFD assay. The test results indicated that, the NASBA-LFD assay showed sensitivity for 1.0 fg M. rosenbergii total RNA with MrNV-chin infection, which was 104 times that of the present RT-PCR approach for the detection of MrNV. In addition, no products were created for shrimps with infection of other kinds of either DNA or RNA virus, which indicated that the NASBA-LFD was specific for MrNV. Therefore, the combination of NASBA and LFD is a new alternative detection method for MrNV which is rapid, accurate, sensitive and specific without expensive equipment and specialised personnel. Early detection of this infectious disease among aquatic organisms will help implement efficient therapeutic strategy to prevent its spread, enhance animal health and limit loss of aquatic breeds in the event of an outbreak.

Distinctive modulation of hepcidin in cancer and its therapeutic relevance.

Hepcidin, a short peptide synthesized primarily by hepatocytes in response to increased body iron and inflammation, is a crucial iron-regulating factor. Hepcidin regulates intestinal iron absorption and releases iron from macrophages into plasma through a negative iron feedback mechanism. The discovery of hepcidin inspired a torrent of research into iron metabolism and related problems, which have radically altered our understanding of human diseases caused by an excess of iron, an iron deficiency, or an iron disparity. It is critical to decipher how tumor cells manage hepcidin expression for their metabolic requirements because iron is necessary for cell survival, particularly for highly active cells like tumor cells. Studies show that tumor and non-tumor cells express and control hepcidin differently. These variations should be explored to produce potential novel cancer treatments. The ability to regulate hepcidin expression to deprive cancer cells of iron may be a new weapon against cancer cells.

Assessment of the benefits of essential fatty acids and risks associated with antimicrobial residues in aquatic products: A case study of Chinese mitten crab (Eriocheir sinensis).

Much attention has been given to the safety and quality of aquatic products, including consuming Chinese mitten crab (Eriocheir sinensis), which offers both nutritional benefits and toxicological risks. Eighteen sulfonamides, 9 quinolones and 37 fatty acids were analyzed in 92 crab samples from primary aquaculture provinces in China. Enrofloxacin and ciprofloxacin have been mentioned as typical antimicrobials occurring at the greatest concentrations (>100 µg/kg, wm). By use of an in vitro method, the proportions of enrofloxacin, ciprofloxacin and essential fatty acids (EFAs, DHA and EPA) in ingested nutrients were determined to be 12 %, none and 95 %, respectively. The risk-benefit quotient (HQ) between the adverse effects of antimicrobials and nutritional benefits of EFAs in crabs found that HQs based on data after digestion were significantly less (HQ = 0.0086) than that of the control group where no digestion occurred (HQ = 0.055). This result suggested that (1) there was less risk posed by antimicrobials due to the consumption of crab, and (2) ignoring the bioaccessible fraction of antimicrobials in crabs might overestimate risks to the health of humans due to dietary exposure. Meaning bioaccessibility can improve the accuracy of the risk assessment process. Realistic risk evaluation should be recommended to achieve a quantified assessment of the dietary risks and benefits of aquatic products.

Enhancing Milk Production by Nutrient Supplements: Strategies and Regulatory Pathways.

The enhancement of milk production is essential for dairy animals, and nutrient supplements can enhance milk production. This work summarizes the influence of nutrient supplements-including amino acids, peptides, lipids, carbohydrates, and other chemicals (such as phenolic compounds, prolactin, estrogen and growth factors)-on milk production. We also attempt to provide possible illuminating insights into the subsequent effects of nutrient supplements on milk synthesis. This work may help understand the strategy and the regulatory pathway of milk production promotion. Specifically, we summarize the roles and related pathways of nutrients in promoting milk protein and fat synthesis. We hope this review will help people understand the relationship between nutritional supplementation and milk production.

Biodegradation and metabolic pathway of sulfamethoxazole by Sphingobacterium mizutaii.

Sulfamethoxazole (SMX) is the most commonly used antibiotic in worldwide for inhibiting aquatic animal diseases. However, the residues of SMX are difficult to eliminate and may enter the food chain, leading to considerable threats on human health. The bacterial strain Sphingobacterium mizutaii LLE5 was isolated from activated sludge. This strain could utilize SMX as its sole carbon source and degrade it efficiently. Under optimal degradation conditions (30.8 °C, pH 7.2, and inoculum amount of 3.5 × 107 cfu/mL), S. mizutaii LLE5 could degrade 93.87% of 50 mg/L SMX within 7 days. Four intermediate products from the degradation of SMX were identified and a possible degradation pathway based on these findings was proposed. Furthermore, S. mizutaii LLE5 could also degrade other sulfonamides. This study is the first report on (1) degradation of SMX and other sulfonamides by S. mizutaii, (2) optimization of biodegradation conditions via response surface methodology, and (3) identification of sulfanilamide, 4-aminothiophenol, 5-amino-3-methylisoxazole, and aniline as metabolites in the degradation pathway of SMX in a microorganism. This strain might be useful for the bioremediation of SMX-contaminated environment.

Detection of cyprinid herpesvirus 2 by loop-mediated isothermal amplification in combination with a lateral flow dipstick.

We developed a convenient technique to detect Herpesviral haematopoietic necrosis attributed to cyprinid herpes virus 2 (CyHV-2), a serious disease of Crucian carp and goldfish related to high mortality. In the present study, we employed a lateral flow dipstick (LAMP-LFD) to present a loop-mediated isothermal amplification assay. The specificity was ascertained via other six viruses, and the sensitivity was compared using PCR method, which are the reaction conditions changes for the method improved. The results revealed that CyHV-2 performance was observable at 64 °C in a separated tube within 60 min, when the samples hybridized using an FITC-labeled probe. As the LAMP-LFD method's specificity was high, with its sensitivity identical to that of traditional PCR, the overall DNA collected revealed the lowest detection limit of 0.18 pg/µl from goldfish diseased by CyHV-2. In summary, the development of LAMP-LFD's method does not require expensive instruments, and it can be regarded as a fast, simple, and reliable method for CyHV-2 detection.

A universal random DNA amplification and labeling strategy for microarray to detect multiple pathogens of aquatic animals.

Diseases caused by bacteria, fungi, and viruses pose a great threat to aquaculture. As DNA microarrays can be used to detect multiple pathogens, here we reported an array with the potential to simultaneously detect 13 bacterial and 11 viral pathogens of aquatic animals. The array included 853 oligonucleotide probes (20-40 mer) complementary to various virus-specific sequences and four chromosomal loci (16S rRNA, gyrB, dnaJ, and recA) of bacteria. Multiplex PCR, phi29 DNA polymerase, and a Klenow fragment-based method were evaluated for amplifying and labeling the nucleic acid of pathogens. While array hybridization signals were most intense using pathogen sequences amplified by multiplex PCR, the phi29 DNA polymerase method was more convenient and ideal since it did not require sequence-specific primers that could bias against detecting novel pathogens. The feasibility of the phi29 DNA polymerase-based microarray strategy was also demonstrated by detecting multiple unknown pathogens from four samples of diseased fish and shrimps.

Root Hair Single Cell Type Specific Profiles of Gene Expression and Alternative Polyadenylation Under Cadmium Stress.

Transcriptional networks are tightly controlled in plant development and stress responses. Alternative polyadenylation (APA) has been found to regulate gene expression under abiotic stress by increasing the heterogeneity at mRNA 3'-ends. Heavy metals like cadmium pollute water and soil due to mining and industry applications. Understanding how plants cope with heavy metal stress remains an interesting question. The Arabidopsis root hair was chosen as a single cell model to investigate the functional role of APA in cadmium stress response. Primary root growth inhibition and defective root hair morphotypes were observed. Poly(A) tag (PAT) libraries from single cell types, i.e., root hair cells, non-hair epidermal cells, and whole root tip under cadmium stress were prepared and sequenced. Interestingly, a root hair cell type-specific gene expression under short term cadmium exposure, but not related to the prolonged treatment, was detected. Differentially expressed poly(A) sites were identified, which largely contributed to altered gene expression, and enriched in pentose and glucuronate interconversion pathways as well as phenylpropanoid biosynthesis pathways. Numerous genes with poly(A) site switching were found, particularly for functions in cell wall modification, root epidermal differentiation, and root hair tip growth. Our findings suggest that APA plays a functional role as a potential stress modulator in root hair cells under cadmium treatment.

The development and application of a duplex reverse transcription loop-mediated isothermal amplification assay combined with a lateral flow dipstick method for Macrobrachium rosenbergii nodavirus and extra small virus isolated in China.

White tail disease (WTD), a major disease prevailing in the larval stage of Macrobrachium rosenbergii, caused by Macrobrachium rosenbergii nodavirus (MrNV) associated with extra small virus (XSV), led to the economic loss of shrimp industry in China. In order to establish a convenient, sensitive and selective molecular diagnostic method to detect MrNV and XSV for the Chinese shrimp (MrNV/XSV-chin), a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay combined with a lateral flow dipstick (LFD) method were developed. A set of four specific primers and a labeled probe were designed according to the six conserved gene sequence regions encoding for the MrNV capsid protein CP43 and the XSV capsid protein CP17. The detection of MrNV and XSV simultaneously by RT-LAMP was performed at 61 °C in a single reaction for 60 min followed by hybridization with an FITC-labeled probe for 5 min and visualized by LFD. The RT-LAMP-LFD assay had a sensitivity of approximately 100-fold higher than conventional PCR. In addition, the assay could detect MrNV/XSV-chin from limited amount of RNA extracts as low as 1.0 pg extracted from Macrobrachium rosenbergii. This assay was simple to use, required little instrumentation, and exhibited excellent specificity for the MrNV/XSV-chin compared with other shrimp viruses. In conclusion, a convenient, sensitive and selective practical molecular diagnostic method was developed with the potential for diagnosis and prevention of WTD.

Draft Genome Sequence of Aeromonas hydrophila Strain BSK-10 (Serotype O97), Isolated from Carassius carassius with Motile Aeromonad Septicemia in China.

We report here a draft genome sequence of Aeromonas hydrophila strain BSK-10, belonging to serotype O97, isolated from crucian carp (Carassius carassius) with motile aeromonad septicemia in Zhejiang, China. The assembly resulted in 34 scaffolds totaling approximately 4.97 Mb, with an average G+C content of 60.97% and 4,594 predicted coding genes.

Isolation and Characterization of a Novel Dicistrovirus Associated with Moralities of the Great Freshwater Prawn, Macrobrachium rosenbergii.

The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important crustacean and is farmed in many countries. Since 2009, a larval mortality syndrome of M. rosenbergii has broken out and spread widely in the main breeding area, including Zhejiang, Jiangsu, Guangxi, and Guangdong Provinces in mainland China. A novel virus, named Macrobrachium rosenbergii Taihu virus (MrTV), was isolated from the moribund larvae and was determined to be the causative agent of the M. rosenbergii larval mortality syndrome by experimental infection. Further genomic sequencing suggested that the MrTV genome is monopartite, 10,303 nt in length, and dicistronic with two non-overlapping open reading frames (ORFs) separated by an intergenic region (IGR) and flanked by untranslated regions (UTRs). Phylogenetic analysis using the full-length genomic sequence and the putative amino acid sequences of the capsid protein revealed that MrTV was more closely related to the taura syndrome virus (TSV) than to any other viruses. According to these molecular features, we proposed that MrTV is a new species in the genus Aparavirus, family Dicistroviridae. These results may shed light on controlling larval mortality syndrome in M. rosenbergii.

Complete Genome Sequence of the Highly Virulent Aeromonas schubertii Strain WL1483, Isolated from Diseased Snakehead Fish (Channa argus) in China.

We sequenced the complete genome of the highly virulent Aeromonas schubertii strain WL1483, which was isolated from diseased snakehead fish (Channa argus) in China. The full genome sequence of A. schubertii WL1483 is 4,400,034 bp, which encodes 4,376 proteins and contains 195 predicted RNA genes.

[Inactivated vaccine for hemorrhage of Grass carp up-regulates the expressions of major immune-related genes in spleen of Grass carp].

OBJECTIVE: To investigate the impact of the injection of inactivated vaccine for hemorrhage of Grass carp on the expressions of main immune-related genes in spleen including immunoglobulin M (IgM), major histocompatibility complexI(MHC I), interferon I(IFN1), complement 3 (C3), interleukin-1ß (IL-1ß) and intelectin genes. METHODS: Ctenopharyngoden idellus kidney (CIK) cells were inoculated with Grass carp reovirus (GCRV). Then inactivated vaccine was prepared by inactivating virus suspension with formaldehyde. Vaccine was 50- and 100-times diluted with normal saline. The experiments included four groups: undiluted group, 50-times diluted group, 100-times diluted group and normal saline control group. Then healthy Grass carps with the average body mass (20 ± 5) g were intraperitoneally injected with vaccine or normal saline separately for each group. Spleens were collected at different time points after injection (0, 6, 12, 24, 72 hours) and total RNA extraction was performed immediately. Real-time quantitative PCR (qRT-PCR) was used to detect mRNA expressions of the six immune-related genes. RESULTS: Compared with normal saline control group, inactivated vaccine injection increased mRNA expressions of the six immune-related genes in the spleen of Grass carp, indicating that the vaccine stimulated the Grass carp to generate non-specific and specific immune responses. In the immunized groups, relative expressions of the genes intelectin, MHCI, IL-1ß, C3 were all up-regulated at first and then declined as time went on. Relative expression of IFNIgene reached the peak at 6 hours post-injection and then decreased gradually to the normal level. Differently, relative expression of IgM gene increased continuously within 0-72 hours post-injection. CONCLUSION: The expressions of the six major immune-related genes were all up-regulated in the spleen of Grass carp injected with different doses of inactivated vaccine for hemorrhage of Grass carp.

[Molecular mechanisms of quinolone resistance in Aeromonas hydrophilia].

OBJECT: To study the resistance mechanisms to quinolones in Aeromonas hydrohila isolated from aquatic animals. METHODS: The drug-resistant spectrum of 23 strains was determined. Quinolone-resistance determining regions of gyrA and parC genes in both screened and in-vitro induction drug-resistant strains were analyzed. Then the detection of quinolone drugs relative efflux pump genes qepA, oqxA and mdfA was performed. The qnrA, qnrB, qnrC, qnrD and qnrS genes were also analyzed at the same time. RESULTS: All organisms were resistant to more than 5 drugs; 39.1% (9/23) of the isolates were quinolone resistant, of which 55.6% (5/9) were enrofloxacin resistant. All the enrofloxacin-resistant isolates harbored qnrS gene, but none of the enrofloxacin-resistant strains harbored qnrA, qnrB, qnrC , qnrD genes and the efflux pump genes of qepA, oqxA and mdfA. AH19 possessed the gyrA and parC genes double mutation, plasmid-mediated quinolone resistance gene qnrS and efflux pump, 3 drug resistance mechanisms simultaneously, while the two drug-resistant mechanisms of AH4, AH7 and AH20 were gyrA and parC genes double mutation and qnrS gene. GyrA gene mutation and qnrS gene occurred in AH6. Compared to the strain ATCC7966, the in-vitro induction drug-resistant strain ATCC7966-QR had both the gyrA and parC genes mutation. CONCLUSION: The mechanisms of resistance to quinolones in the A. hydrophila isolates of this study mainly depended on the existence of plasmid-mediated gene qnrS and the variation of the target site of quinolone drugs, whereas, the drug resistance mechanism relying on the efflux pump system only existed in individual strains.