Heat stable antifungal factor (HSAF) is considered to be a potential biological pesticide due to its broad antifungal activity and novel mode of action. However, few studies have reported on HSAF ...production during fermentation. Thus, this work was executed to optimize the medium composition to maximize HSAF production by Lysobacter enzymogenes OH11, with soybean flour, glucose and CaCl2 identified as suitable nutrients with concentrations of 8·00, 7·89 and 0·72 g l−1 respectively. Simultaneously, the quantitative analysis of HSAF production was established by eliminating the emulsification problem, and the highest HSAF production was determined to be 356·34 ± 13·86 mg l−1 using the optimized medium, 12‐fold higher than when using the 10% TSB medium (29·34 ± 2·57 mg l−1). Furthermore, the cost of this medium was assessed and nearly 31‐fold lower than that of 10% TSB. This study suggests that the optimized medium is not only effective but also economical for HSAF production.
Significance and Impact of the Study
Significance and Impact of the Study: Heat stable antifungal factor (HSAF) exhibits a potent and broad antifungal activity with a novel mode of action. Increased production and reduced cost of raw materials are particularly important for the future production of HSAF, however, no report was involved in these studies. This study aimed to improve the production of HSAF with cheap raw materials through the medium optimization, which would lay the foundation for the application of HSAF in biological control.
Significance and Impact of the Study: Heat stable antifungal factor (HSAF) exhibits a potent and broad antifungal activity with a novel mode of action. Increased production and reduced cost of raw materials are particularly important for the future production of HSAF, however, no report was involved in these studies. This study aimed to improve the production of HSAF with cheap raw materials through the medium optimization, which would lay the foundation for the application of HSAF in biological control.
Glucose is the primary precursor for the synthesis of lactose, which controls milk volume by maintaining the osmolarity of milk. Glucose uptake in the mammary gland plays a key role in milk ...production. Glucose transport across the plasma membranes of mammalian cells is carried out by 2 distinct processes: facilitative transport, mediated by a family of facilitative glucose transporters (GLUT); and sodium-dependent transport, mediated by the Na+/glucose cotransporters (SGLT). Transport kinetic studies indicate that glucose transport across the plasma membrane of the lactating bovine mammary epithelial cell has a K(m) value of 8.29 mM for 3-O-methyl-D-glucose and can be inhibited by both cytochalasin-B and phloretin, indicating a facilitative transport process. This is consistent with the observation that in the lactating bovine mammary gland, GLUT1 is the predominant glucose transporter. However, the bovine lactating mammary gland also expresses GLUT3, GLUT4, GLUT5, GLUT8, GLUT12, and sodium-dependent SGLT1 and SGLT2 at different levels. Studies of protein expression and cellular and subcellular localizations of these transporters are needed to address their physiological functions in the mammary gland. From late pregnancy to early lactation, expression of GLUT1, GLUT8, GLUT12, SGLT1, and SGLT2 mRNA increases from at least 5-fold to several hundred-fold, suggesting that these transporters may be regulated by lactogenic hormones and have roles in milk synthesis. The GLUT1 protein is detected in lactating mammary epithelial cells. Its expression level decreases from early to late lactation stages and becomes barely detectable in the nonlactating gland. Both GLUT1 mRNA and protein levels in the lactating mammary gland are not significantly affected by exogenous bovine growth hormone, and, in addition, GLUT1 mRNA does not appear to be affected by leptin.
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•A total of 17 antibiotics were detected in water, sediment (biofilm), organism and feed samples.•Greenhouse ponds were more likely to accumulate antibiotics compared to outdoor ...ponds.•Zn as the coexisting contaminant was significantly correlated with antibiotic concentrations.•Aquaculture wastewater and domestic sewage were potential sources of antibiotics.•Ciprofloxacin and enrofloxacin may present high ecological and resistance risks.
This study focused on the distribution, combined pollution, potential source and risk assessment of 17 antibiotics in an aquaculture ecosystem surrounding the Yellow sea, North China. Antibiotics were detected in various matrices (seawater, sediment/biofilm, organism and feed) in different aquaculture modes (greenhouse and outdoor aquaculture) during the wet and dry seasons in coastal areas of Shandong province. The innovation points of the study were as follows: (1) To the best of our knowledge, this study was one of the few to investigate the occurrence and distribution of antibiotics in mariculture environments along the Yellow Sea coast; (2) Biofilms, a focus of the study, might act as a sink for antibiotics in the aquaculture ecosystem; and (3) The correlation of heavy metals and antibiotic concentrations was proved, which could correspondingly be used as an indicator for antibiotic concentrations in the studied area.
The levels of antibiotics in water were observed to be relatively low, at the ng/L level. Trimethoprim was the most prevalent antibiotic, and was detected in all water samples. Oxytetracycline was detected at high concentrations in biofilms (up to 1478.29 ng/g). Moreover, biofilms exhibited a higher antibiotic accumulation capacity compared to sediments. Concentrations of oxytetracycline and doxycycline were high in feed, while other antibiotics were almost undetected. Tetracycline was widely detected and the concentration of enrofloxacin was highest in organisms.
Correlation analysis demonstrated that environmental parameters and other coexisting contaminants (e.g. heavy metals) significantly affected antibiotic concentrations. In addition, the concentration of Zn was significantly correlated with the total antibiotic concentration and was proportional to several antibiotics in water and sediment (biofilm) samples (p < 0.01). High Mn concentrations were closely related to total and individual (e.g. sulfadiazine, sulfamethazine and enrofloxacin) antibiotic levels, which may result in the combined contamination of the environment. Antibiotics in estuaries and groundwater generally originated from aquaculture wastewater and untreated/treated domestic sewage. Most of the detected antibiotics posed no risk to the environment. Ciprofloxacin and enrofloxacin found in water may present high ecological and resistance risks, while the two antibiotics observed to accumulate in fish may pose a considerable risk to human health through diet consumption. All antibiotics detected in seafood were lower than the respective maximum residue limits. This study can act as a reference for the government for the determination of antibiotic discharge standards in aquaculture wastewater and the establishment of a standardized antibiotic monitoring and management system.
When dairy cows are exposed to high-temperature environment, their antioxidant capacity and productive performance decrease, leading to economic losses. Emerging evidence has shown that selenium (Se) ...can effectively alleviate heat stress in dairy cows; however, the cellular mechanism underlying this protection is not clear. The purpose of this study was to investigate and compare the protective effects of inorganic Se (sodium selenite, SS) and organic Se (selenite methionine, SM) in MAC-T (mammary alveolar cells-large T antigen, a bovine mammary epithelial cell (BMEC) line) cells during heat stress. MAC-T cells were treated in 4 ways unless otherwise described: (i) cells in the heat treatment (HT) group were cultured at 42.5°C for 1 h and then recovered in 37°C for another 12 h; (ii) the SM group was pretreated with organic Se for 2 h, cultured at 42.5°C for 1 h, and then recovered in 37°C for 12 h; (iii) the SS group was treated similarly to the SM group except that the cells were pretreated with inorganic Se instead of organic Se; and (iv) the control group was continuously cultured in 37°C and received no Se treatment. The results showed that heat shock at 42.5°C for 1 h triggered heat shock response, sabotaged the redox balance, and reduced cell viability in MAC-T cells; and pretreatment of cells with SM or SS effectively alleviated the negative effects of heat shock on the cells. However, the cells were much more sensitive to SS treatment but more tolerant to SM. In addition, two forms of Se appeared to affect the expression of different genes, including nuclear factor erythroid 2-related factor 2 (Nrf2) and inducible nitric oxide synthase (iNOS) in the SM group and thioredoxin reductase 1 (TXNRD1) in the SS group in Nrf2-ARE (antioxidant response element) antioxidant pathway and inflammation response. In summary, results showed the mechanistic differences in the protective effects of organic and inorganic Se on heat stress in BMECs.
Gastric cancer (GC) is one of the most common tumors and the molecular mechanism underlying its metastasis is still largely unclear. Here, we show that miR-25 was overexpressed in plasma and primary ...tumor tissues of GC patients with tumor node metastasis stage (III or IV) or lymph node metastasis. MiR-25 inhibition significantly decreased the metastasis, invasion and proliferation of GC cells in vitro, and reduced their capacity to develop distal pulmonary metastases and peritoneal dissemination in vivo. Furthermore, miR-25 repressed transducer of ERBB2, 1 (TOB1) expression by directly binding to TOB1-3'-UTR, and the inverse correlation was observed between the expressions of miR-25 and TOB1 mRNA in primary GC tissues. Moreover, the loss of TOB1 increased the metastasis, invasion and proliferation of GC cells, and the restoration of TOB1 led to suppressed metastasis, invasion and proliferation. The receiver operating characteristics analysis yielded an area under the curve value of 0.7325 in distinguishing the GC patients with death from those with survival. The analysis of optimal cutoff value revealed poor survival in GC patients with high plasma concentrations of miR-25 (>0.2333 amol/μl). Taken together, miR-25 promotes GC progression by directly downregulating TOB1 expression, and may be a noninvasive biomarker for the prognosis of GC patients.
Nuclear factor (erythroid-derived 2)-like factor 2 (Nrf2) is a transcription factor that binds to the antioxidant response element (ARE) in the upstream promoter region of many antioxidative genes. ...The Nrf2-ARE signaling plays a key role in the cellular antioxidant-defense system, but whether Nrf2 activation has protective effects against heat shock (HS) stress in mammary epithelial cells (MEC) remains unclear. The objective of this study was to determine whether tert-butylhydroquinone (tBHQ), a well-known Nrf2 activator, could attenuate heat stress-induced cell damage in MAC-T cells of the bovine MEC line. The MAC-T cells were exposed to HS (42.5°C for 1h) followed by recovery at 37°C to mimic HS. Compared with cells that were consistently cultured at normothermia (37°C), the cell viability levels significantly decreased after HS stress. In parallel, heat stress increased the reactive oxygen species levels and induced cellular apoptosis and endoplasmic reticulum stress. The MAC-T cells that were pretreated with tBHQ (10μM) for 2h followed by HS had a reduction in the loss of cell viability. The tBHQ pretreatment significantly decreased cellular reactive oxygen species levels and stress-related marker gene expression. The tBHQ-treated MAC-T cells showed strong Nrf2-ARE signaling activation and a nuclear accumulation of Nrf2 and upregulated expression of Nrf2-ARE downstream genes. Small interfering RNA silencing of Nrf2 in HS-treated MAC-T cells almost completely abolished the cytoprotective effects by tBHQ. Overall, our results demonstrated that HS could cause cell damage in cultured bovine MEC, and that activation of Nrf2 by tBHQ could attenuate HS-induced cell damage.
The spatiotemporal distribution, source apportionment and combined pollution of 14 antibiotics in natural waters adjacent to mariculture farms of Laizhou Bay in the Bohai Sea were studied. The ...contribution proportion and quantity of each potential pollution source to antibiotics in natural water bodies were quantitatively described. The correlations between heavy metals and antibiotics and their underlying mechanisms in natural and aquaculture water environment were analyzed.
Fourteen antibiotics were detected in natural water and sediment in the coastal area of Laizhou Bay. The maximum concentrations of sulfamethazine and trimethoprim in water reached tens or even hundreds of μg/L in winter. Trimethoprim was the main antibiotic in natural water bodies in winter and summer, and enrofloxacin was the principal antibiotic in sediments. Enrofloxacin, ciprofloxacin and oxytetracycline were detected in all underground water samples; thus, control of these antibiotics needs to be made a priority to mitigate groundwater contamination.
PCA–MLR revealed that the potential sources of antibiotics in natural waters of Laizhou Bay include the mariculture wastewater (18.3%), the domestic sewage (63.3%) and the livestock wastewater (18.4%). Therefore, the antibiotic burden of Laizhou Bay was principally from the domestic sewage. In natural water, the concentration of Cu was positively correlated with antibiotics, which might be related to the common sources, the competitive adsorption in sediments and the easy complexation characteristic of Cu and antibiotics. Positive correlations among antibiotics and heavy metals were observed in mariculture sediments, while negative relationships were observed in natural sediments.
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•The main antibiotics in natural water and sediment were TMP and ENR, respectively.•ENR, CIP and OTC were detected in the groundwater surrounding the Laizhou Bay.•Antibiotic burden of seawater in Laizhou Bay was principally from domestic sewage.•As a coexisting contaminant, Cu was positively correlated with some antibiotics.
Infusion of lipopolysaccharides (LPS) into a mammary gland can provoke inflammatory responses and impair lactation in both the infused gland and neighboring glands. To gain insight into the ...mechanisms controlling the spatiotemporal response to localized mastitis in lactating dairy cows, we performed RNA sequencing on mammary tissue from quarters infused with LPS, neighboring quarters in the same animals, and control quarters from untreated animals at 3 and 12 h postinfusion. Differences in gene expression were annotated to Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Comparing mammary transcriptomes from all 3 treatments revealed 3,088 and 1,644 differentially expressed (DE) genes at 3 and 12 h, respectively. Of these genes, >95% were DE only in LPS-infused quarters and represented classical responses to LPS: inflammation, apoptosis, tissue remodeling, and altered cell signaling and metabolism. Although relatively few genes were DE in neighboring quarters (56 at 3 h; 74 at 12 h), these represented several common pathways. At 3 h, tumor necrosis factor (TNF), nuclear factor-κB, and nucleotide-binding and oligomerization domain (NOD)-like receptor signaling pathways were identified by the upregulation of anti-inflammatory (NFKBIA, TNFAIP3) and cell adhesion molecule (VCAM1, ICAM1) genes in neighboring glands. Additionally, at 12 h, several genes linked to 1-carbon and serine metabolism were upregulated. Some responses were also regulated over time. The proinflammatory response in LPS-infused glands diminished between 3 and 12 h, indicating tight control over transcription to re-establish homeostasis. In contrast, 2 glucocorticoid-responsive genes, FKBP5 and ZBTB16, were among the top DE genes upregulated in neighboring quarters at both time points, indicating potential regulation by glucocorticoids. We conclude that a transient, systemic immune response was sufficient to disrupt lactation in neighboring glands. This response may be mediated directly by proinflammatory factors from the LPS-infused gland or indirectly by secondary factors released in response to systemic inflammatory signals.
Iron selenide superconductors exhibit a number of unique characteristics that are helpful for understanding the mechanism of superconductivity in high-Tc iron-based superconductors more generally. ...However, in the case of AxFe2Se2 (A = K, Rb, Cs), the presence of an intergrown antiferromagnetic insulating phase makes the study of the underlying physics problematic. Moreover, FeSe-based systems intercalated with alkali metal ions, NH3 molecules or organic molecules are extremely sensitive to air, which prevents the further investigation of their physical properties. It is therefore desirable to find a stable and easily accessible FeSe-based superconductor to study its physical properties in detail. Here, we report the synthesis of an air-stable material, (Li0.8Fe0.2)OHFeSe, which remains superconducting at temperatures up to ~40 K, by means of a novel hydrothermal method. The crystal structure is unambiguously determined by a combination of X-ray and neutron powder diffraction and nuclear magnetic resonance. Moreover, antiferromagnetic order is shown to coexist with superconductivity. This synthetic route opens a path for exploring superconductivity in other related systems, and confirms the appeal of iron selenides as a platform for understanding superconductivity in iron pnictides more broadly.
MicroRNAs (miRNAs) deregulation is frequent in human gastric cancers (GCs), but the role of specific miRNAs involved in this disease remains elusive. MiR-22 was previously reported to act as tumor ...suppressors or oncogenes in diverse cancers. However, their accurate expression, function and mechanism in GC are largely unclear. Here, we found that the expression of miR-22 was significantly reduced in clinical GC tissues compared with paired adjacent normal tissues, and was significantly correlated with a more aggressive phenotype of GC in patients, and miR-22 low expression correlated with poor overall survival. The introduction of miR-22 markedly suppressed GC cell growth, migration and invasion, and inhibition of miR-22 promoted GC cell proliferation, migration and invasion in vitro. We further demonstrated that miR-22 acted as tumor suppressors through targeting extracellular matrix (ECM) remodeling member matrix metalloproteinase 14 (MMP14) and epithelial-to-mesenchymal transition (EMT) inducer Snail in GC. Moreover, ectopic expression of MMP14 or Snail restored inhibitory effects of miR-22 on cell migration and invasion in GC cells, and a negative relationship between the miR-22 expression and MMP14 or Snail mRNA levels was observed in GC. Finally, overexpression of miR-22 suppressed tumor growth, peritoneal dissemination and pulmonary metastasis in vivo. Taken together, we identified that miR-22 is a potent tumor suppressor in GC. MiR-22 downregulation promotes GC invasion and metastasis by upregulating MMP14 and Snail, and then inducing ECM remodeling and EMT. These findings provide a better understanding of the development and progression of GC and may be an important implication for future therapy of the GC.
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