Viable but non-culturable state formation and resuscitation of different antibiotic-resistant Escherichia coli induced by UV/chlorine.

The presence of "viable but nonculturable" (VBNC) state and bacterial antibiotic resistance (BAR) both pose significant threats to the safety of drinking water. However, limited data was available that explicitly addressed the contribution of bacterial VBNC state in the maintenance and propagation of BAR. Here, the VBNC state induction and resuscitation of two antibiotic-resistant Escherichia coli K12 strains, one carrying multidrug-resistant plasmid (RP4 E. coli) and the other with chromosomal mutation (RIF E. coli) were characterized by subjecting them to different doses of UV/chlorine. The results illustrated that the induction, resuscitation, and associated mechanisms of VBNC ARB exhibit variations based on resistance determinants. RP4 E. coli exhibited a higher susceptibility to enter VBNC state compared to the RIF E. coli., and most VBNC state and resuscitated RP4 E. coli retained original antibiotic resistance. While, reverse mutation in the rpoB gene was observed in VBNC state and recovered RIF E. coli strains induced by high doses of UV/chlorine treatment, leading to the loss of rifampicin resistance. According to RT-qPCR results, ARGs conferring efflux pumps appeared to play a more significant role in the VBNC state formation of RP4 E. coli and the down-regulation of rpoS gene enhanced the speed at which this plasmid-carrying ARB entered into the dormant state. As to RIF E. coli, the induction of VBNC state was supposed to be regulated by the combination of general stress response, SOS response, stringent response, and TA system. Above all, this study highlights that ARB could become VBNC state during UV/chlorine treatments and retain, in some cases, their ability to spread ARGs. Importantly, compared with chromosomal mutation-mediated ARB, both VBNC and resuscitated state ARB that carries multidrug-resistant plasmids poses more serious health risks. Our study provides insights into the relationship between the VBNC state and the propagation of BAR in drinking water systems.

Extracellular vesicles enriched with miR-150 released by macrophages regulates the TP53-IGF-1 axis to alleviate myocardial infarction.

Myocardial infarction (MI) is recognized as a major cause of death and disability around the world. Macrophage-derived extracellular vesicles (EVs) have been reportedly involved in the regulation of cellular responses to MI. Thus, we sought to clarify the mechanism by which macrophage-derived EVs regulate this process. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was performed to determine microRNA-150 (miR-150) expression in an MI mouse model with ligation of the left anterior descending coronary artery (LAD) and in hypoxia/reoxygenation (H/R)-exposed cardiomyocytes. Bioinformatics analysis and dual luciferase reporter gene assay were adopted to identify the correlation of miR-150 with tumor protein 53 (TP53) expression in cardiomyocytes. Gain- and loss-of-function experiments were conducted in H/R-induced cardiomyocytes, cardiomyocytes incubated with EVs from miR-150 mimic-transfected macrophages, or MI-model mice treated with EVs from miR-150 mimic-transfected macrophages. hematoxylin-eosin (HE) and terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining assays were used for detecting inflammatory infiltration and cell apoptosis. The release of lactate dehydrogenase (LDH) by dead cardiomyocytes was measured with an LDH kit, and the apoptosis-related proteins, Bax, and cleaved-caspase 3 were determined by Western blot analysis. miR-150 expression was downregulated in the infarcted cardiac tissues of MI mice. Macrophage-derived EVs could transfer miR-150 into cardiomyocytes, where it directly targeted and suppressed TP53. Furthermore, miR-150 suppressed phosphatase and tensin homology (PTEN) and activated p-Akt to upregulate IGF-1 expression. Furthermore, increased expression of EV-derived miR-150 prevented cardiomyocyte apoptosis in vitro, as evidenced by downregulated Bax and cleaved-caspase 3 and upregulated Bcl2 and alleviated MI in vivo. In conclusion, our study demonstrates the cardioprotective effect of macrophage-derived EV-miR-150 on MI-induced heart injury through negatively regulating the TP53-IGF-1 signaling pathway.NEW & NOTEWORTHY miR-150 is expressed at a low level in cardiac tissues after myocardial infarction. Macrophages-derived EVs transfer miR-150 to cardiomyocytes. miR-150 directly targets TP53. miR-150 elevation regulates TP53-IGF-1 axis to reduce cardiomyocyte apoptosis. EV-derived miR-150 could be a potential therapeutic target for myocardial infarction.

Effects of Organic Copper on Growth Performance and Oxidative Stress in Mice.

Copper (Cu) has been used as a feed additive for many years. However, high Cu amounts can cause oxidative stress and adversely affect animal performance. Such negative effects may depend on the amounts and forms of Cu. In the present study, the effects of inorganic Cu (CuSO4) and organic Cu (chelate-Cu) present in mice feed on daily growth rate and Cu deposition in the liver, kidneys, spleen, brain, and serum were assessed in addition to the oxidative stress levels in the liver and brain. Organic Cu at a concentration of 15 mg/kg significantly enhanced daily growth rate in mice, whereas Cu deposition in the livers was significantly lower than that in the inorganic Cu group. Glutathione peroxidase activity in the liver of the mice fed with organic Cu significantly improved, whereas malondialdehyde levels in the brain and liver were significantly lower than that in the inorganic Cu group. The different effects of organic Cu and inorganic Cu provide key evidence supporting the use of organic Cu in animal feeds.

Comparative proteomics analysis of myocardium in mouse model of diabetic cardiomyopathy using the iTRAQ technique.

BACKGROUND: Diabetic cardiomyopathy (DCM) is one of the most harmful diseases with high morbidity and mortality rates. However, the underlying pathological mechanism of the disorder still remains unclear. OBJECTIVES: The purpose of our study was to identify differentially expressed proteins associated with DCM. MATERIAL AND METHODS: C57BLKS/J db/db (diabetes mellitus group - DM group) and db/m mice (normal control group - NC group) were acclimated in cages for 15 weeks. The general state was recorded. After 15 weeks, the heart tissues were used for histological examination. In addition, quantitative mass spectrometry using isobaric tags for relative and absolute quantitation (iTRAQ) was used to identify differentially expressed proteins in the heart tissues. SEQUEST software was used to identify proteins with data derived from liquid chromatography-tandem mass spectrometry (LC-MS/MS) spectra by searching ipi.MOUSE.v3.72.REVERSED. fasta database. Expert Protein Analysis System (ExPASy) was used to calculate the theoretical parameters. One upregulated protein (sorbin and SH3 domain containing 2 - Sorbs2) and 1 downregulated protein (myosin-3) was measured by western blot to validate the iTRAQ data. RESULTS: The mice in the NC group were active and grew well, while the mice in the DM group presented with obvious polydipsia, polyphagia and polyuria. The results of histological examination revealed that, compared to the NC group, the DM group showed significant myocardial hypertrophy and myofiber disarray accompanied by damaged nuclei. A total of 73 differentially expressed proteins were identified, including 44 upregulated and 29 downregulated proteins. Western blot analysis confirmed that the expression of Sorbs2 was significantly increased (p < 0.01), while the expression of myosin-3 was statistically decreased in the DM group compared to the NC group (p < 0.05). CONCLUSIONS: These results suggest that DCM shows differences in its proteomics compared to normal controls. Our quantitative proteomic analysis may provide a new insight into the distinct molecular profile of DCM.

A Non-Targeted Liquid Chromatographic-Mass Spectrometric Metabolomics Approach for Association with Coronary Artery Disease: An Identification of Biomarkers for Depiction of Underlying Biological Mechanisms.

BACKGROUND We performed non-targeted metabolomics analysis using liquid chromatography-mass spectrometry coupled technique to explore the biological mechanism of coronary artery disease (CAD) events for improved prediction. MATERIAL AND METHODS We studied the association of CAD events in 4092 individuals and observed the replication of sphingomyelin (28:1), lysophosphatidylcholine (18:2), lysophosphatidylcholine (18:1), and monoglyceride (18:2), which were independent of main CAD risk factors. RESULTS We found that these 4 metabolites were responsible for traditional risk factors and also contributed to the modifications related to reclassification and discrimination. Monoglycerides (MonoGs) were positively associated with C-reactive proteins and body mass index, while lysophosphatidylcholines (LPPCs), which had less evidence of subclinical CAD in an additional 1010 participants, yielded a reverse pattern. An association between monoGs and CAD independence of triglycerides (triGs) were also observed. On the basis of Mendelian randomization analysis, we observed a positive but weak irregular effect (odds ratio per unit increase in standard deviation in monoG=1.11, P-value=0.05) on CAD. CONCLUSIONS Our work establishes the relationship of metabolome with coronary artery disease and explains the biological mechanism of CAD events, as we identified the above-mentioned metabolites along with the evidence supporting their clinical use.

Cardioprotective effect of indirubin in experimentally induced myocardial infarction in wistar rats.

Recently, there has been as enhanced interest on global level to recognize the potent antioxidant compounds which are pharmacologically active with less or no side effects. Thus, the current investigation was intended to scrutinize the protective effect of indirubin on the cardiac marker, such as, enzymes, LDH isoenzyme, cardiac troponin-T (cTnT), antioxidant enzymes marker and lipid peroxidation (LPO) in response of isoproterenol (ISO)-induced myocardial infarction (MI) in Wistar rats. The experimental animals were categorized into following groups: Group I received saline; Group II received Indirubin (10 mg/kg); Group III received ISO (100 mg/kg) and Group IV received ISO + indirubin (10 mg/kg) for continuous 10 days. The ISO induced MI injury was confirmed via enhanced level of enzymes markers viz., creatine kinase-MB, creatine kinase, lactate dehydrogensae, troponin-T, alanine transaminase (ALT) and aspartate transaminase (AST) in the rats serum. The enhanced expression of LDH (1 and 2) isoenzyme bands were also observed in the ISO induced MI rats. We have also estimated the level of LPO in the heart and plasma, which was found to be significantly (P<0.05) improved. Moreover, the marker of enzymatic antioxidant enzymes viz., glutathione reductase (GRx), catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPx) and superoxide dismutase (SOD) in the heart, and the level of non-enzymatic antioxidant marker viz., vitamin (C, E) in heart and serum were found to be considerably (P<0.05) reduced in the ISO induced MI in Wistar rats. Whereas, the ISO control Wistar rats showed significant (P<0.05) increase in the uric acid level in the plasma. The Indirubin treated rats confirmed the significant protective effect via modulation of all biological and antioxidant parameters tested. The result of the investigation was further found in agreement of the histopathological studies of the indirubin treated rats which clearly showed recovery from the myocardial infarction. Thus, on the basis of that, it has been suggested that indirubin showed protection of myocardial tissues against the ISO persuaded oxidative stress.