Coculture of Acinetobacter johnsonii and Shewanella putrefaciens Contributes to the ABC Transporter that Impacts Cold Adaption in the Aquatic Food Storage Environment.

Acinetobacter johnsonii and Shewanella putrefaciens were identified as specific spoilage organisms in aquatic food. The interactions among specific spoilage organisms under cold stress have a significant impact on the assembly of microbial communities, which play crucial roles in the spoilage and cold adaptation processes. The limited understanding of A. johnsonii and S. putrefaciens interactions in the cold adaptation mechanism hinders the elucidation of their roles in protein and metabolism levels. 4D quantitative proteomic analysis showed that the coculture of A. johnsonii and S. putrefaciens responds to low temperatures through ABC transporter proteins, resulting in phospholipid transport and inner membrane components. SapA and FtsX proteins were significantly upregulated, while LolC, LolD, LolE, PotD, PotA, PotB, and PotC proteins were significantly downregulated. Metabolome assays revealed that metabolites of glutathione and spermidine/putrescin were significantly upregulated, while metabolites of arginine/lysine/ornithine were significantly downregulated and involved in the ABC transporter metabolism. The results of ultramicroscopic analyses showed that the coculture of A. johnsonii and S. putrefaciens surface combined with the presence of the leakage of intracellular contents, suggesting that the bacteria were severely damaged and wrinkled to absorb metabolic nutrients and adapt to cold temperatures.

Applications of Genomics, Metabolomics, Fourier Transform Infrared in the Evaluation of Spoilage Targets of Shewanella putrefaciens from Spoiled Bigeye Tuna.

Shewanella putrefaciens is a typical spoiler that is commonly found in seafood and has high spoilage potential. However, the spoilage mechanism against Shewanella putrefaciens at the gene and metabolism levels has not been well elucidated. This work determined the spoilage targets on Shewanella putrefaciens XY07 from spoiled bigeye tuna by genome sequencing, metabolomics, and Fourier transform infrared (FTIR) analysis. Shewanella putrefaciens XY07 contained some genes on spoilage regulating of cys genes, his genes, spe genes and rpoS gene involved in sulfur metabolism, histidine metabolism, arginine and proline degradation, and biofilm formation at the genome level, respectively. Some spoilage genes like speC, cysM, trxB genes were identified. In addition, ABC transporters, arginine and proline metabolism; beta-alanine metabolism; glycine, serine, and threonine metabolism; histidine metabolism; sulfur metabolism; and lipid metabolism were identified as important pathways related to aquatic food during spoilage, which indicated the functions of amino acid degradation in S. putrefaciens XY 07 by metabolomics analysis. The metabolites of l-ornithine, 5-aminopentanoate, and 4-aminobutyraldehyde could be further metabolized to spermidine and spermine, producing a spoilage odor, and were involved in arginine and proline metabolism serving as key spoilage regulating metabolisms. Therefore, Shewanella putrefaciens XY07 was applied to genomics, metabolomics analysis, and FTIR to provide comprehensive insight into the investigation of spoilage targets.

Response to Cold Adaption in Acinetobacter johnsonii XY27 from Spoiled Bigeye Tuna (Thunnus obesus): Membrane Protein Composition and Protein Biomarker Identification by Proteomics.

Acinetobacter johnsonii is one of the major food-spoilage bacteria and can survive under cold stress. In this study, the membrane composition, membrane permeability, and energy transduction of A. johnsonii XY27 cultured at 4 and 30 °C were examined comparatively by flow cytometry combined with liquid chromatography-mass spectrometry and gas chromatography-mass spectrometry. The Na+/K+ATPase activity, alkaline phosphatase and ATPase activity, fluorescence intensity, and cell viability in A. johnsonii XY27 increased with the decrease in cultivation temperature. The polyunsaturated fatty acid and monounsaturated fatty acids have a higher content in A. johnsonii XY27 cultured at 4 °C compared to that cultured at 30 °C, in which the contents of methyl palmitoleate, methyl myristoleate, and methyl oleate increased dramatically with decreasing temperature. Comparative proteomics analysis revealed that 31 proteins were downregulated and 4 proteins were upregulated, in which catalase-peroxidase 1 and cold shock proteins as biomarker proteins could effectively control A. johnsonii during cold adaptation.

SNHG9/miR-214-5p/SOX4 feedback loop regulates osteosarcoma progression.

Osteosarcoma (OS) is a high-grade, aggressive bone sarcoma. LncRNAs play a key regulatory role in controlling biological and pathological processes. The expression of lncRNA SNHG9 varies among different cancer tissues, and the role of SNHG9 in OS progression is unclear. In this study, we found SNHG9 overexpression in OS tissues and cells. In addition, downregulated SNHG9 expression impaired the proliferation, migration, and invasion abilities of OS cells. SNHG9 expression was positively regulated by the transcription factor SOX4. SNHG9 interacted with miR-214-5p as a molecular sponge and SOX4 was identified as the target of miR-214-5p. The interaction affected the expression of SNHG9, miR-214-5p, and SOX4, and regulated OS cell proliferation, migration, and invasion. Therefore, the SNHG9/miR-214-5p/SOX4 feedback loop performs an important role in OS progression and might be used as a new potential therapeutic target for the treatment of OS.

Comparison of Physicochemical Changes and Water Migration of Acinetobacter johnsonii, Shewanella putrefaciens, and Cocultures From Spoiled Bigeye Tuna (Thunnus obesus) During Cold Storage.

This study investigates the physicochemical changes and water migration of Acinetobacter johnsonii (A), Shewanella putrefaciens (S), and cocultured A. johnsonii and S. putrefaciens (AS) inoculated into bigeye tuna during cold storage. The physicochemical indexes [fluorescence ratio (FR), total volatile base nitrogen (TVB-N), thiobarbituric acid (TBA), trimethylamine (TMA), peroxide value (POV), and pH] of bigeye tuna increased cold storage. A significant decrease in trapped water was found in the AS samples, and direct monitoring of the water dynamics was provided by low-field nuclear magnetic resonance. Samples inoculated with A. johnsonii and S. putrefaciens also induced the degradation of myofibrillar proteins and weakness of some Z-lines and M-lines. Higher values of physicochemical indexes and water dynamics were shown in the coculture of S. putrefaciens and A. johnsonii than in the other groups. Therefore, this paper reveals that the coculture of A. johnsonii and S. putrefaciens resulted in a bigeye tuna that was more easily spoiled when compared to the single culture. This study provides insight into the spoilage potential of A. johnsonii and S. putrefaciens during cold storage, which further assists in the application of appropriate technologies to keep the freshness of aquatic foods.

Differences in lipid composition of Bigeye tuna (Thunnus obesus) during storage at 0 °C and 4 °C.

This study aimed to investigate the lipid oxidation and distribution in Bigeye tuna stored at 0 °C and 4 °C for 6 days. Tuna were evaluated by determining the peroxide value (POV), acid value (AV), anisidine value (AnV), polyene index, fluorescence ratio (FR), phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) content, and major glycerophospholipid molecular species. The value of lipid oxidation indexes (POV, AV, AnV, FR, PC, PE and PI) increased as the storage time increased. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) results indicated that the major types of lipids included diacylglycerol (DAG), monoacylglycerol (MAG), phospholipid (PL), and triacylglycerol (TAG). At least 136 PC and 64 PE molecular species were identified in Bigeye tuna. The results of the confocal laser scanning microscope analysis indicated the distribution of TAG and PL particles. In addition, principal component analysis showed that the contents of PI and TAG were positively correlated with PC, polyene index and lipid content but negatively correlated with PI, POV, FR, AOV, AnV, MAG, and DAG, which might be explained by distinguishing the lipid parameters affecting lipid oxidation. Therefore, this study may provide a novel method to evaluate lipid changes and contribute to the balanced nutritional value of aquatic foods during cold storage.

Applications of Non-invasive and Novel Methods of Low-Field Nuclear Magnetic Resonance and Magnetic Resonance Imaging in Aquatic Products.

Aquatic products, such as fish, are popular throughout the world due to their satisfying flavor characteristics as well as rich animal nutrition, and they provide high-value food therapy, but they are easily oxidized and spoiled. It is necessary to detect aquatic products through rapid and accurate technology. Low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) have been widely used in the aquatic product industry due to their sensitivity, fast analysis, non-destructive nature and low cost. The applications of LF-NMR in the measurement of aquatic product quality and nutrients (water, fat, and protein) are summarized in this paper. Applications in aquatic products have been shown to depend on deep processing, storage and authentication. This review discusses the application of MRI technology in the quality control of aquatic products. Therefore, this review will guide the application of the aquatic products industry and aims to supply the reader with both the theory of the method and practical applications of the method for use as a rapid and non-destructive technology in scientific research and the industrial industry.

Quorum Sensing System-Regulated Proteins Affect the Spoilage Potential of Co-cultured Acinetobacter johnsonii and Pseudomonas fluorescens From Spoiled Bigeye Tuna (Thunnus obesus) as Determined by Proteomic Analysis.

Food spoilage by certain species of bacteria is reported to be regulated by quorum sensing (QS). Acinetobacter johnsonii and Pseudomonas fluorescens, the major specific spoilage organisms, are found to be limited in their QS and co-culture interactions. The aim of this study was to determine how QS-regulated proteins affect the spoilage potential of co-cultured A. johnsonii and P. fluorescens obtained from spoiled bigeye tuna (Thunnus obesus) using a proteomics approach. The A. johnsonii, P. fluorescens, and their co-culture tested the N-acyl-homoserine lactone (AHL) activities using reporter Chromobacterium violaceum CV026 and LC-MS/MS in qualitative and quantitative approaches, respectively. These latter showed that, of the 470 proteins and 444 proteins in A. johnsonii (A) and P. fluorescens (P), respectively, 80 were significantly up-regulated and 97 were significantly down-regulated in A vs. AP, whereas 90 were up-regulated and 65 were down-regulated in P vs. AP. The differentially expressed proteins included the AI-2E family transporter OS, 50S ribosomal protein L3, thioredoxin reductase OS, cysteine synthase CysM OS, DNA-binding response regulator, and amino acid ABC transporter ATPase OS. The cellular process (GO:0009987), metabolic process (GO:0008152), and single-organism process (GO:0044699) were classified into the gene ontology (GO) term. In addition, energy production and conversion, amino acid transport and metabolism, translation, ribosomal structure and biogenesis, post-translational modification, protein turnover, and chaperones were distributed into the clusters of orthologous groups of proteins (COG) terms. The KEGG pathways revealed that 84 and 77 differentially expressed proteins were divided into 20 KEGG pathways in A vs. AP and P vs. AP, respectively, and amino acid metabolism, carbohydrate metabolism, energy metabolism, and translation were significantly enriched. Proteins that correlated with the spoilage-related metabolic pathways, including thioredoxin reductase OS, cysteine synthase OS, and pyridoxal phosphate-dependent enzyme family protein OS, were identified. AI-2E family transporter OS and LuxR family transcriptional regulator OS were identified that related to the QS system. These findings provide a differential proteomic profile of co-culture in A. johnsonii and P. fluorescens, and have potential applications in QS and the regulation of spoilage potential.

Growth Kinetics and Spoilage Potential of Co-culturing Acinetobacter johnsonii and Pseudomonas fluorescens from Bigeye Tuna (Thunnus obesus) During Refrigerated Storage.

A. johnsonii and P. fluorescens are the well-known specific spoilage organisms in aquatic products and the study of the interactions between A. johnsonii and P. fluorescens are limited. This study aims to evaluate the growth kinetics, spoilage potential and interactions of A. johnsonii and P. fluorescens isolated from spoiled bigeye tuna (Thunnus obesus) by inoculating into sterile fish slices and stored at 4 °C for 6 days. The growth kinetics of A. johnsonii and P. fluorescens were fitted with Baranyi and Roberts model. The chemical indexes (total volatile base nitrogen (TVB-N), trimethylamine (TMA), pH, proteolytic activity and protein content) of each inoculated block of bigeye tuna were increased during refrigerated storage. Moreover, the higher contents of chemical indexes were observed in co-culture with A. johnsonii and P. fluorescens compared with single culture of A. johnsonii and P. fluorescens. In addition, atomic force microscopy (AFM) observation of co-culturing A. johnsonii and P. fluorescens inoculation into sterile fish slices revealed damage of myofibrillar protein structures and the protein degradation. Based on these parameters, a rapid method to evaluate spoilage potential of A. johnsonii and P. fluorescens was positively correlated with TVB-N value, TMA value and pH value (P < 0.05) by the correlation coefficient. Consequently, spoilage potential of microorganisms became stronger evaluated in a mixed culture than single culture. This paper provides insight for a detection method of interactions of A. johnsonii and P. fluorescens during refrigerated storage.

A non-invasive method for quantitative monitoring of quality changes and water migration in bigeye tuna (Thunnus obesus) during simulated cold chain logistics using low-field nuclear magnetic resonance coupled with PCA.

Bigeye tuna (Thunnus obesus) is an important fish species worldwide due to its desirable flavour and rich nutrition. Cold chain logistics is one of the most common methods to store and sell bigeye tuna (Thunnus obesus). The purpose of this study was to investigate how bigeye tuna (T. obesus) deteriorate during simulated cold chain logistics with fluctuating temperatures by monitoring the quality changes and water distribution. The physical and chemical results showed that the sensory score, texture profile analysis (TPA), and water holding capacity (WHC) decreased, while total volatile basic nitrogen (TVB-N), K value, aerobic plate counts (APC), and psychrotrophic bacterial counts increased with fluctuating temperatures during 132 h. Low-field nuclear magnetic resonance (LF-NMR) and magnetic resonance imaging (MRI) detection showed that the T21 (trapped water) and proton density decreased, while T22 (free water) decreased initially and then increased, indicating continuous water loss and water migration during storage. Principal component analysis (PCA) revealed that springiness, hardness, chewiness, T21, and sensory score were correlated with each other (p < 0.05) and the linear combination of quality indicators and T2transverse relaxation time was established. Besides, temperature fluctuation (batch 3) accelerated the deterioration of bigeye tuna according to the comprehensive score of PCA. Results were based on LF-NMR which can be used to monitor quality changes in a fast non-invasive manner.

Study on the Volatile Organic Compounds and Its Correlation with Water Dynamics of Bigeye Tuna (Thunnus obesus) during Cold Storage.

Volatile organic compounds (VOCs) and water play a key role in evaluating the quality of aquatic products. Quality deterioration of aquatic products can produce some off-odour volatiles and can induce water content changes. However, no previous study has investigated a correlation between water dynamics and VOCs of bigeye tuna during cold storage. The changes in VOCs, water dynamics and quality attributes of bigeye tuna (Thunnus obesus) upon storage at 0 °C and 4 °C for 6 days were investigated. The results showed that the values of ATP, adenosine diphosphate (ADP), adenosine monophosphate (AMP), T21 (trapped water) and the relative value of T1 decreased (p < 0.05), while drip loss and histamine contents increased (p < 0.05), which indicated quality deterioration during cold storage. With haematoxylin and eosin (HE) staining, muscle tissue microstructure was observed. VOCs such as hexanal, heptanal, 4-Heptenal, (Z)-, pentadecanal-, 1-pentanol, 1-hexanol significantly increased, which sharply increased the content of off-odour volatiles. T21 was positively correlated with 1-octen-3-ol, 1-penten-3-ol, while T21 was negatively correlated with hexanal, 1-hexanol. Therefore, good correlations between water dynamics and some VOCs were detected during quality deterioration of bigeye tuna throughout cold storage.

Effects of liraglutide on hemodynamic parameters in patients with heart failure.

Glucagon-like peptide-1 analogues improve left ventricular function in patients with acute myocardial infarction. This study aimed to evaluate the effects of liraglutide on hemodynamic parameters in patients with heart failure. A total of 78 patients with heart failure were enrolled in this study between August 2014 and November 2015. Of these, 52 patients were randomized 1:1 to receive either liraglutide or placebo for 7 days. Hemodynamic measurements were made using transpulmonary thermodilution and arterial pulse contour analysis. At 7 days, the difference in change of the primary endpoint of cardiac output between the liraglutide group and control group was +1.1 1/min (95% CI +0.1 to +2.2; P < 0.001). Stroke volume was significantly higher in the liraglutide group compared with the control group (difference: +14.6 ml; P < 0.001). The difference in an increase in the left ventricular contractile index after 7 days of treatment was +210.7 mmHg/s (liraglutide versus control, 95% CI-92.1 to +501.5; P < 0.001). Liraglutide causes favorable changes in markers of inflammation and oxidative stress. Glucagon-like peptide-1 may be associated with improvement in left ventricular function in patients with heart failure. These findings need to be confirmed by larger invasive trials.

China national lung cancer screening guideline with low-dose computed tomography (2015 version).

BACKGROUND: Lung cancer is the leading cause of cancer-related death in China. Results from a randomized controlled trial using annual low-dose computed tomography (LDCT) in specific high-risk groups demonstrated a 20% reduction in lung cancer mortality. METHODS: A China national lung cancer screening guideline was developed by lung cancer early detection and treatment expert group appointed by the National Health and Family Planning Commission, based on results of the National Lung Screening Trial, systematic review of evidence related to LDCT screening, and protocol of lung cancer screening program conducted in rural China. RESULTS: Annual lung cancer screening with LDCT is recommended for high risk individuals aged 50-74 years who have at least a 20 pack-year smoking history and who currently smoke or have quit within the past five years. Individualized decision making should be conducted before LDCT screening. LDCT screening also represents an opportunity to educate patients as to the health risks of smoking; thus, education should be integrated into the screening process in order to assist smoking cessation. CONCLUSIONS: A lung cancer screening guideline is provided for the high-risk population in China.

[Effects of cutting and reseeding on the ground-dwelling arthropod community in Caragana intermedia forest in desert steppe].

Taking a 25-year-old Caragana intermedia forest in desert steppe as test object, an investigation was conducted on the ground-dwelling arthropod community in cutting and no-cutting stands with and without reseeding, aimed to understand the effects of cutting, reseeding and their interaction on the individual number and group richness of ground-dwelling arthropod in C. intermedia forest. There were significantly lower number and richness of ground-dwelling arthropod in the open spaces than under the shrubs in the no-cutting and no-reseeding stands. Cutting, reseeding and both of them could significantly increase the number and richness of ground-dwelling arthropod in the open spaces, but not under the shrubs, compared with no cutting or reseeding. Consequently, there were no significant differences in the distribution of ground-dwelling arthropod in the open spaces and under the shrubs in the cutting, reseeding, or cutting and reseeding stands. Further, there was a similar buffer effect between cutting and reseeding on the ground-dwelling arthropod. No significant differences were observed in the ground-dwelling arthropod distribution, between cutting stand and reseeding stand, between cutting stand and cutting and reseeding stand, and between reseeding stand and cutting and reseeding stand. It was suggested that cutting, reseeding, or both of them could significantly improve the ground-dwelling arthropod diversity especially in the open spaces, being beneficial for the restoration of degraded grassland ecosystem and the rational management on artificial C. intermedia forest in desert steppe.

Coniferous canopy BRF simulation based on 3-D realistic scene.

It is difficulties for the computer simulation method to study radiation regime at large-scale. Simplified coniferous model was investigated in the present study. It makes the computer simulation methods such as L-systems and radiosity-graphics combined method (RGM) more powerful in remote sensing of heterogeneous coniferous forests over a large-scale region. L-systems is applied to render 3-D coniferous forest scenarios, and RGM model was used to calculate BRF (bidirectional reflectance factor) in visible and near-infrared regions. Results in this study show that in most cases both agreed well. Meanwhile at a tree and forest level, the results are also good.

Expression and significance of Survivin mRNA in lung cancer tissue microarray detected by FISH.

OBJECTIVE: To investigate the expression of Survivin mRNA in lung cancer tissue microarray (TMA) by fluorescence in situ hybridization (FISH) method, and determine the role and significance of it in lung cancer genesis and progress. METHODS: The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. RESULTS: Survivin mRNA was expressed in 66.7% (36/54) of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue (0/10; chi2 = 15.238, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer (20/24, 83.3%) than moderate and well differentiated cancer (16/30, 53.3%; chi2 = 5.40, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis (27/32, 84.4%) than without lymph node metastasis (9/22, 40.9%; chi2 = 11.084, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in stage III-IV(12/13, 92.3%) than stage I - II (24/41, 58.5%; chi2 = 5.066, P < 0.05). CONCLUSION: Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment.

[Expression of KAI1, MRP-1, and FAK proteins in lung cancer detected by high-density tissue microarray].

BACKGROUND & OBJECTIVE: The genesis, development, invasion, and metastasis of tumor are closely correlate with alterations of multi-genes. At present, the roles of Kang-ai-1 (KAI1), motility-related protein-1 (MRP-1), and focal adhesion kinase (FAK) in lung cancer have seldom been reported. This study was designed to investigate the roles of KAI1, MRP-1, and FAK in tumorigenesis and development of lung cancer, and their values in diagnosis and predicting the prognosis of lung cancer. METHODS: The expression of KAI1, MRP-1, and FAK proteins in a high-density tissue microarray containing 240 spots were detected by SP immunohistochemistry. RESULTS: The positive rates of KAI1 and MRP-1 were significantly lower in primary lung cancer than in normal lung tissue (25.9% vs. 100%, 42.6% vs. 100%, P<0.05). The positive rate of FAK was significantly higher in primary lung cancer than in normal lung tissue (44.4% vs. 10.0%, P<0.05). The expression of KAI1, FAK, and MRP-1 in primary lung cancer had no correlation with age and gender of the patients, and macroscopic and histological type of tumor, but had correlations with tumor differentiation, clinical stage, and lymph node metastasis. In addition, the expression of MRP-1 had correlation with histological type of tumor; the positive rate of MRP-1 was significantly lower in small cell lung cancer than in non-small cell lung cancer (0 vs. 50.0%, P<0.05). KAI1 expression was negatively correlated to FAK expression (rs=-0.458, P<0.05); MRP-1 expression was positively correlated with KAI1 expression (rs=0.535, P<0.05), and negatively correlated with FAK expression (rs=-0.438, P<0.05). CONCLUSIONS: The abnormal expression of KAI1, MRP-1, and FAK proteins are related to invasion and metastasis of lung cancer. Combined detection of the 3 proteins may be useful in predicting the development and prognosis of lung cancer.