Recovery of Cold Impacted Escherichia Coli O157:H7 Bacteria in Ground Beef

Escherichia coli in fresh minced meat was injured by cooling at 4 °C. A bacterial population has three different physiology which are uninjured or normal cells, sublethally injured cells (or injured cells ), and lethally injured cells (or dead cells). Cell injury is defined as any damage to the components of cells themselves by any stresses which weaken the ability of cells to survive or multiply. This will increase the sensitivity of cells to any harmful factors. The cells can repair their injury which can be extended 48hour depending on the nature of stress and degree of injury. The purpose of this study was to: supplemented some cultural media and preparation new cultural media to isolated E.coli with compounds that supplemented the bacterial growth such as yeast extract, sodium pyruvate, n-propyl gallate, catalase, and Tween. Various concentrations of the compound were tested minced beef meat with mixed it and compared with traditional media. The rest of the compound had variable effects on the recovery of cold stressed cells but they weren’t as efficient as needed. It is, therefore recommended that 0.5% of both catalase and tween 80 be used to supplement tryptic soy agar (TSA) in the repair detection procedure.

Evolving Escherichia coli Host Strains for Efficient Deuterium Labeling of Recombinant Proteins Using Sodium Pyruvate-d3

Labeling of proteins with deuterium (2H) is often necessary for structural biology techniques, such as neutron crystallography, NMR spectroscopy, and small-angle neutron scattering. Perdeuteration in which all protium (1H) atoms are replaced by deuterium is a costly process. Typically, expression hosts are grown in a defined medium with heavy water as the solvent, which is supplemented with a deuterated carbon source. Escherichia coli, which is the most widely used host for recombinant protein production, can utilize several compounds as a carbon source. Glycerol-d8 is often used as a carbon source for deuterium labelling due to its lower cost compered to glucose-d7. In order to expand available options for recombinant protein deuteration, we investigated the possibility of producing a deuterated carbon source in-house. E. coli can utilize pyruvate as a carbon source and pyruvate-d3 can be made by a relatively simple procedure. To circumvent the very poor growth of E. coli in minimal media with pyruvate as sole carbon source, adaptive laboratory evolution for strain improvement was applied. E. coli strains with enhanced growth in minimal pyruvate medium was subjected to whole genome sequencing and the genetic changes were revealed. One of the evolved strains was adapted for the widely used T7 RNA polymerase overexpression systems. Using the improved strain E. coli DAP1(DE3) and in-house produced deuterated carbon source (pyruvic acid-d4 and sodium pyruvate-d3), we produce deuterated (>90%) triose-phosphate isomerase, at quantities sufficient enough for large volume crystal production and subsequent analysis by neutron crystallography.

Recording the Presence of Peanibacillus larvae larvae Colonies on MYPGP Substrates Using a Multi-Sensor Array Based on Solid-State Gas Sensors

American foulbrood is a dangerous disease of bee broods found worldwide, caused by the Paenibacillus larvae larvae L. bacterium. In an experiment, the possibility of detecting colonies of this bacterium on MYPGP substrates (which contains yeast extract, Mueller-Hinton broth, glucose, K2HPO4, sodium pyruvate, and agar) was tested using a prototype of a multi-sensor recorder of the MCA-8 sensor signal with a matrix of six semiconductors: TGS 823, TGS 826, TGS 832, TGS 2600, TGS 2602, and TGS 2603 from Figaro. Two twin prototypes of the MCA-8 measurement device, M1 and M2, were used in the study. Each prototype was attached to two laboratory test chambers: a wooden one and a polystyrene one. For the experiment, the strain used was P. l. larvae ATCC 9545, ERIC I. On MYPGP medium, often used for laboratory diagnosis of American foulbrood, this bacterium produces small, transparent, smooth, and shiny colonies. Gas samples from over culture media of one- and two-day-old foulbrood P. l. larvae (with no colonies visible to the naked eye) and from over culture media older than 2 days (with visible bacterial colonies) were examined. In addition, the air from empty chambers was tested. The measurement time was 20 min, including a 10-min testing exposure phase and a 10-min sensor regeneration phase. The results were analyzed in two variants: without baseline correction and with baseline correction. We tested 14 classifiers and found that a prototype of a multi-sensor recorder of the MCA-8 sensor signal was capable of detecting colonies of P. l. larvae on MYPGP substrate with a 97% efficiency and could distinguish between MYPGP substrates with 1–2 days of culture, and substrates with older cultures. The efficacy of copies of the prototypes M1 and M2 was shown to differ slightly. The weighted method with Canberra metrics (Canberra.811) and kNN with Canberra and Manhattan metrics (Canberra. 1nn and manhattan.1nn) proved to be the most effective classifiers.

Sodium Pyruvate Ameliorates Influenza a Virus Infection In Vivo

Influenza A virus (IAV) causes seasonal epidemics annually and pandemics every few decades. Most antiviral treatments used for IAV are only effective if administered during the first 48 h of infection and antiviral resistance is possible. Therapies that can be initiated later during IAV infection and that are less likely to elicit resistance will significantly improve treatment options. Pyruvate, a key metabolite, and an end product of glycolysis, has been studied for many uses, including its anti-inflammatory capabilities. Sodium pyruvate was recently shown by us to decrease inflammasome activation during IAV infection. Here, we investigated sodium pyruvate’s effects on IAV in vivo. We found that nebulizing mice with sodium pyruvate decreased morbidity and weight loss during infection. Additionally, treated mice consumed more chow during infection, indicating improved symptoms. There were notable improvements in pro-inflammatory cytokine production (IL-1β) and lower virus titers on day 7 post-infection in mice treated with sodium pyruvate compared to control animals. As pyruvate acts on the host immune response and metabolic pathways and not directly on the virus, our data demonstrate that sodium pyruvate is a promising treatment option that is safe, effective, and unlikely to elicit antiviral resistance.

65 Co-culture of porcine epithelial oviductal cells and invitro-produced embryos: Effect on embryo development and quality

The oviduct is involved in many reproductive functions, including early embryo development. The epithelial cells that cover the oviduct produce oviducal fluid and could be used to recreate the invivo environment into which embryo development takes place. This study aimed to evaluate the co-culture of porcine embryos with a monolayer of porcine oviducal epithelial cells (POEC) and its effect on embryo development and quality. The POEC were obtained by pressing the isthmus (from diestrus sow oviducts) using slides and performing 3 cycles of vortexing and decanting in DMEM-F12 medium. Passage 1 cells were used for these experiments (POEC-1). Oocytes were obtained from follicular aspiration of slaughterhouse ovaries. Oocytes were invitro matured for 44h in TCM-199 supplemented with human menopausal gonadotrophin and cyclic AMP during the first 22h. Invitro fertilization was performed with 17°C-refrigerated boar semen for 4h in 100-µL drops of TCM-199 with caffeine, bovine serum albumin, sodium lactate, and sodium pyruvate (20 denuded oocytes per drop, 1×106 spermatozoa mL−1). Presumptive zygotes were washed and randomly assigned to one of the following groups for invitro culture: control (50-µL drop of NCSU-23 with sodium pyruvate and lactate), POEC-1 (same as the control+POEC-1 50 000 cells mL−1), POEC-1+FBS (same as the control+POEC-1 50 000 cells mL−1 and 2.5% of fetal bovine serum). Culture conditions were 7% O2, 5% CO2, 39°C, and humidity. On Day 2, the cleavage rate was recorded, and embryos were transferred to NCSU-23 drops with glucose and without cells. The blastocyst rate was recorded on Day 7. Embryo quality was assessed by counting the number of cells per blastocyst (Hoechst) and the apoptosis index (TUNEL-positive cells/total cells). Co-culture with POEC-1 significantly increased the blastocyst rate (control: 14%; POEC-1+FBS: 10%; POEC-1: 28%; P<0.05 Chi-squared test) and allowed embryo hatching (control: 0; POEC-1+FBS: 22.2%; POEC-1: 7; P<0.05 Chi-squared test). However, there was no significant difference in the number of cells per blastocyst (control: 58.6±6; POEC-1+FBS: 50.3±3.7; POEC-1: 50.6±4.8; nonparametric ANOVA) or in the apoptosis index (control: 8.1; POEC+FBS 8.3; POEC: 7.4; nonparametric ANOVA). The use of POEC-1 during the first 2 days of embryo culture enhanced embryo development and improved culture conditions, allowing embryo hatching. The effect on embryo development could be due to an effect of POEC itself or the effect of feeder cells. Other parameters of embryo quality should be evaluated in the future.

Biocompatible photoinduced CuAAC using sodium pyruvate

Sodium pyruvate, a natural intermediate produced during cellular metabolism, is commonly used in buffer solutions and media for biochemical applications. Here we show the use of sodium pyruvate (SP) as...

NAD+, Senolytics, or Pyruvate for Healthy Aging?

In last decades, healthy aging has become one of research hotspots in life science. It is well known that the nicotinamide adenine dinucleotide oxidized form (NAD+) level in cells decreases with aging and aging-related diseases. Several years ago, one of NAD+ precursors was first demonstrated with its new role in DNA damage repairing in mice, restoring old mice to their physical state at young ones. The finding encourages extensive studies in animal models and patients. NAD+ and its precursors have been popular products in nutrition markets. Alternatively, it was also evidenced that clearance of cellular senescence by senolytics preserved multiorgan (kidney and heart) function and extended healthy lifespan in mice. Subsequent studies confirmed findings in elderly patients subjected with idiopathic pulmonary fibrosis. The senolytic therapy is now focused on various diseases in animal and clinical studies. However, pyruvate, as both a NAD+ substitute and a new senolytic, may be advantageous, on the equimolar basis, over current products above in preventing and treating diseases and aging. Pyruvate-enriched fluids, particularly pyruvate oral rehydration salt, may be a novel intervention for diseases and aging besides critical care. Albeit the direct evidence that benefits healthy aging is still limited to date, pyruvate, as both NAD+ provider and senolytic agent, warrants intensive research to compare NAD+ or senolytics for healthy aging, specifically on the equimolar basis, in effective blood levels. This review briefly discussed the recognition of healthy aging by comparing NAD+ and Senolytics with sodium pyruvate from the clinical point of view.