Addition Of Sodium Pyruvate Research Articles

Role of sodium pyruvate in maintaining the survival and cytotoxicity of Staphylococcus aureus under high glucose conditions.

Glucose is a crucial carbon source for the growth of Staphylococcus aureus, but an excess of glucose is detrimental and even leads to cell death. Pyruvate, the central metabolite of glycolysis, has been shown to have anti-inflammatory and antioxidant properties. This study aimed to investigate the protective effect of pyruvate on S. aureus under high glucose conditions. Sodium pyruvate greatly increased the cytotoxicity of S. aureus strain BAA-1717 to human erythrocytes and neutrophils in vitro. However, the cytotoxicity and survival of S. aureus were significantly reduced by high glucose, which was restored to normal levels by the addition of sodium pyruvate. The expression of hlg and lukS in S. aureus was higher in the LB-GP cultures than that in LB-G cultures, but there was no significant difference in cytotoxicity between LB-GP and LB-G cultures. Furthermore, the hemolytic activity of S. aureus supernatants could be inhibited by the cell-free culture medium (CFCM) of LB-G cultures, suggesting that high levels of extracellular proteases were presence in the CFCM of LB-G cultures, resulting in degradation of the hemolytic factors. The expression of sarA, which negatively regulates extracellular protease secretion, was higher in LB-GP cultures than that in LB-G cultures. Additionally, sodium pyruvate increased acetate production in S. aureus, which helps maintain cell viability under acidic environment. In conclusion, pyruvate plays an important role in the survival and cytotoxicity of S. aureus under high glucose conditions. This finding may aid in the development of effective treatments for diabetic foot infections.

Scaling-Up an Aqueous Self-Degassing Electrochemically Mediated ATRP in Dispersion for the Preparation of Cellulose-Polymer Composites and Films.

Electrochemically mediated atom transfer radical polymerization (eATRP) is developed in dispersion conditions to assist the preparation of cellulose-based films. Self-degassing conditions are achieved by the addition of sodium pyruvate (SP) as a ROS scavenger, while an aluminum counter electrode provides a simplified and more cost-effective electrochemical setup. Different polyacrylamides were grown on a model cellulose substrate which was previously esterified with 2-bromoisobutyrate (-BriB), serving as initiator groups. Small-scale polymerizations (15 mL) provided optimized conditions to pursue the scale-up up to 1000 mL (scale-up factor ~67). Cellulose-poly(N-isopropylacrylamide) was then chosen to prepare the tunable, thermoresponsive, solvent-free, and flexible films through a dissolution/regeneration method. The produced films were characterized by Fourier-transform infrared (FTIR), scanning electron microscopy (SEM), dynamic scanning calorimetry (DSC), and thermogravimetric analysis (TGA).

Media impacts recovery of Salmonella enterica and Enterococcus faecium NRRL B2354 from whole black peppercorns, basil leaves, and chia seeds treated with antimicrobial gasses

Salmonella enterica contamination of low water activity foods (LWAFs) has resulted in recalls of spices, herbs, and seeds and outbreaks of salmonellosis. To improve the safety of these ready-to-eat products, new treatment methods, including fumigation with chlorine dioxide (ClO2) or hydrogen peroxide (H2O2) gas are being explored, and effectiveness determined. To prevent overestimation of treatment effectiveness, it is vital that recovery methods should accurately quantify all viable cells, even those injured. This study evaluated different media and supplements for the recovery of multiple strains of S. enterica and Enterococcus faecium NRRL B2354, from ClO2 or H2O2 treated black peppercorns, dried basil leaves, and chia seeds. Also, this study aimed to compare the log reduction of these two microorganisms to evaluate E. faecium NRRL B2354, as a surrogate for S. enterica. On average, recovery of S. enterica was improved by 1 log CFU from ClO2 and H2O2 treated LWAFs when a non-selective but differential media containing tryptic soy agar with yeast extract, ammonium iron citrate and sodium thiosulfate (MTSAYE) was used, when compared to plating on XLD (p < 0.05). Furthermore, addition of sodium pyruvate, ferrous sulfate, or 3’3’-thiodiproionate supplements to MTSAYE did not show increased recovery of either S. enterica or E. faecium NRRL B2354 (p > 0.05). On each treatment and LWAF combination tested, there was no significant difference between the log reduction of S. enterica and E. faecium NRRL B2354, indicating its suitability as a surrogate under the test conditions.

Pyruvate prevents the onset of the cachectic features and metabolic alterations in myotubes downregulating STAT3 signaling.

Cachexia is a systemic disease associated with several pathologies, including cancer, that leads to excessive weight loss due to enhanced protein degradation. Previously, we showed that cachectic features in myotubes are provoked by a metabolic shift toward lactic fermentation. Our previous results led us to hyphotesise that increasing pyruvate concentration could impede the metabolic modifications responsible for induction of cachexia in myotubes. Here, we demonstrated that the addition of sodium pyruvate in conditioned media from CT26 colon cancer cells (CM CT26) prevents the onset of either phenotypic and metabolic cachectic features. Myotubes treated with CM CT26 containing sodium pyruvate show a phenotype similar to the healthy counterpart and display lactate production, oxygen consumption, and pyruvate dehydrogenase activity as control myotubes. The use of the Mitochondrial Pyruvate Carrier inhibitor UK5099, highlights the importance of mitochondrial pyruvate amount in the prevention of cachexia. Indeed, UK5099-treated myotubes show cachectic features as those observed in myotubes treated with CM CT26. Finally, we found that sodium pyruvate is able to decrease STAT3 phosphorylation level, a signaling pathway involved in the induction of cachexia in myotubes. Collectively, our results show that cachexia in myotubes could be prevented by the utilization of sodium pyruvate which impedes the metabolic modifications responsible for the acquisition of the cachectic features.

Media composition to revive leptospires stored at −80 °C

Leptospires are preserved by frequent sub-culturing in semisolid media due to the challenge of low recovery by freezing or liquid nitrogen methods. The present study evaluated three liquid EMJH medium compositions (Medium A: Leptospira medium base EMJH, Leptospira enrichment EMJH, 5-fluorouracil (3%), rabbit serum (1%) and calf serum (1%); Medium B: same as Medium A but without 5-fluorouracil; Medium C: same as Medium B but with the addition of sodium pyruvate) for the revival of leptospires after storage at −80 °C. A total of 18 Leptospira serovars cultured in Medium A was aliquoted into cryogenic vials and directly stored at −80 °C. A hundred microlitre from each serovar culture stored at −80 °C was sub-cultured on a selected time over a period of 30 months into Media A, B and C. Regrowth on Media B and C showed a better and faster recovery (89–100%) (p-value <0.05) compared to Medium A (67–100%). Leptospires can be stored longer at −80 °C and a good recovery could be obtained when sub-cultured on EMJH medium without 5-fluorouracil.

43 Effect of addition of sodium pyruvate in extender on post-thaw quality of Beetal buck semen.

43 Effect of addition of sodium pyruvate in extender on post-thaw quality of Beetal buck semen.

Induction of a viable but non-culturable state in Salmonella Typhimurium by thermosonication and factors affecting resuscitation.

The objective of this work was to analyze the effects of thermosonication (TS) on induction of a viable but non-culturable (VBNC) state in Salmonella Typhimurium and to examine incubation factors affecting subsequent resuscitation of cells. A TS treatment of 380 W at 53°C for 30 min induced the VBNC state in S. Typhimurium cells in beef peptone yeast (BPY) broth, apple/carrot juice, physiological saline and phosphate buffer solution. The logarithmic and decline phases of growth were more sensitive to the TS treatment compared to stationary phase cells. Meanwhile, VBNC S. Typhimurium could be resuscitated back to culturable cells by using suitable incubation temperatures and media. Addition of Tween 20 hindered resuscitation compared to the use of BPY medium alone. The optimal growth temperature (i.e. 37°C) was the most suitable temperature to resuscitate cells from the VBNC state. The VBNC incidence index decreased with the addition of sodium pyruvate during TS treatment, as it accelerated resuscitation. The results demonstrated that free radicals produced during TS processing and the growth phase of cells affected induction of the VBNC state in S. Typhimurium. Hence, the kinds and amounts of free radicals generated during TS treatment should be analyzed in the future.

Improving coenzyme Q8 production in Escherichia coli employing multiple strategies.

Coenzyme Q (CoQ) is a medically valuable compound and a high yielding strain for CoQ will have several benefits for the industrial production of CoQ. To increase the CoQ(8) content of E. coli, we blocked the pathway for the synthesis of menaquinone by deleting the menA gene. The blocking of menaquinone pathway increased the CoQ(8) content by 81 % in E. coli (ΔmenA). To study the CoQ producing potential of E. coli, we employed previous known increasing strategies for systematic metabolic engineering. These include the supplementation with substrate precursors and the co-expression of rate-limiting genes. The co-expression of dxs-ubiA and the supplementation with substrate precursors such as pyruvate (PYR) and parahydroxybenzoic acid (pHBA) increased the content of CoQ(8) in E. coli (ΔmenA) by 125 and 59 %, respectively. Moreover, a 180 % increase in the CoQ(8) content in E. coli (ΔmenA) was realized by the combination of the co-expression of dxs-ubiA and the supplementation with PYR and pHBA. All in all, CoQ(8) content in E. coli increased 4.06 times by blocking the menaquinone pathway, dxs-ubiA co-expression and the addition of sodium pyruvate and parahydroxybenzoic acid to the medium. Results suggested a synergistic effect among different metabolic engineering strategies.

Media Improvement for Detection Freezing Stressed Escherichia coli

Escherichia coli was injured by freezing at-18oC. Surviving cells were recovered on nonselective medium (tryptone glucose extract agar) and selective medium (violet red bile salt agar) supplemented with compounds that degrade hydrogen peroxide or block its formation. Various concentrations of the following compounds were tested: sodium pyruvate, sodium thioglycolate,and n-propyl gallate.Sodium pyruvate and sodium thioglycolate, when added to either medium, significantly increased (P≤0.05) for recovering of injured cells. Supplementation of violet red bile agar increased recovery cells, but the counts remained considerably lower than the tryptone glucose extract agar counts. The repair detection of freezing stressed E.coli was greatly improved by the addition of sodium pyruvate and thioglycolate .We therefore recommend that pyruvate (0.4%) and thioglycolate (0.04%) be used to supplemented media in the repair detection procedure

Influence of growth conditions on pressure resistance of Vibrio parahaemolyticus in oysters and the optimization of postpressure treatment recovery conditions.

Vibrio parahaemolyticus ATCC 43996 was grown at 15°C for 53 h, 20°C for 24 h, 25°C for 12 h, 30°C for 9 h, 35°C for 9 h, or 40°C for 6 h to early stationary phase. Oyster meats were blended, autoclaved at 121°C for 15 min, inoculated with V. parahaemolyticus, and pressure treated at 250 MPa for 2 and 3 min and at 300 MPa for 1 and 2 min at 21°C. Overall, growth temperatures of 20 and 40°C yielded the greatest pressure resistance in V. parahaemolyticus. The effects of salt concentration and H(2)O(2)-degrading compounds on the recovery of V. parahaemolyticus also were investigated. Sterile oyster meats were inoculated with V. parahaemolyticus and treated at 250 MPa for 1, 2, or 3 min at 21°C. These meats were then blended with 0.1% peptone water supplemented with 0.5 to 1.5% NaCl and plated on tryptic soy agar (TSA) supplemented with 0 to 3.5% NaCl. For recovery of pressure-injured cells, peptone water with 1% NaCl and TSA with 0.5% NaCl were the best diluent and plating medium, respectively. Addition of sodium pyruvate (0.05 to 0.2%) or catalase (8 to 32 U/ml) did not increase the recovery of V. parahaemolyticus after pressure treatment. The effect of incubation temperature and gas atmosphere on the recovery of V. parahaemolyticus after pressure treatment also was determined. Aerobic incubation at 30°C resulted in the highest recovery of V. parahaemolyticus in sterile oyster meats. The 30°C incubation temperature was also the optimum temperature for recovery of V. parahaemolyticus in pressure-treated live oysters. The results of this study indicate that the growth conditions for V. parahaemolyticus before and after high hydrostatic pressure treatment should be taken into consideration when assessing the efficacy of pressure inactivation.

Dormant forms of Micrococcus luteus and Arthrobacter globiformis not platable on standard media

The colony-forming ability of long (3–9 months) incubated cystlike resting cells (CRC) of the nonspore-forming gram-positive bacteria Micrococcus luteus and Arthrobacter globiformis was studied in this work. The preservation of the CRC proliferative potential as assayed by plating on standard LB agar was shown to depend on the conditions of the formation of the dormant cells. In aged post-stationary cultures of micrococci and arthrobacters grown under carbon and phosphorus limitation the number of colony-forming units (CFU/ml) of CRC decreased in the course of 3–9 month incubation to the level of 106–107 CFU/ml. However, M. luteus CRC obtained under carbon and nitrogen limitation and A. globiformis CRC obtained under nitrogen limitation and starvation completely lost their ability to form colonies on standard solid medium after 4–6 months of incubation and turned into a ‘non-culturable’ (non-platable) state. In this case, the ratio of live cells in the population of M. luteus and A. globiformis ‘non-culturable’ CRCs (determined by the Live/Dead staining test) was 10–44% of the total cell number. To study the possible preservation of proliferative potential in non-platable CRCs, various methods of their reactivation were applied. Although preincubation of CRC suspensions in a buffer solution of 0.1 M K2HPO4 (pH 7.4) or in the presence of lysozyme (1 or 10 μg/ml) resulted in increased numbers of live cells (determined by the Live/Dead test) or in disruption of the cell conglomerates, it did not increase considerably the CFU titer on LB medium. Variations in the medium composition, such as addition of sodium pyruvate as an antioxidant or dilution of the medium, promoted the formation of macrocolonies by a small portion of nonplateable CRC of M. luteus (50−80 CFU/ml), whereas the number of the cells capable of microcolony formation (mCFU) was 1.8–6.8 × 105 mCFU/ml, exceeding the CFU titers by four orders of magnitude. The application of semisolid agar and the most probable number (MPN) method was the most efficient for determination of the mCFU titer, and an almost complete reversion of ‘non-culturable’ micrococcal CRCs to microcolony formation was observed (up to 2.3 × 107 mCFU/ml). The usefulness of diluted complete media for the restoration of the colony-forming ability of the dormant forms was confirmed in experiments with ‘nonculturable’ CRCs of A. globiformis. The development of special procedures and methods for determining actively proliferating cells not detected by ordinary methods is of great importance for advanced monitoring studies.

Recovery and detection of Vibrio vulnificus during cold storage

Different cultural techniques and molecular methods for the detection of Vibrio vulnificus during cold storage in a model broth system were compared. Two strains of V. vulnificus were grown to stationary phase and inoculated (10 6 CFU/mL) into tryptic soy broth with 2% sodium chloride (TSBN 2) or artificial seawater (ASW), both pre-chilled to 5 °C. These were stored for 10 days, with sub-sampling conducted at time 0 and every 2 days thereafter. Each subsample was plated, by both pour and spread plate techniques, onto tryptic soy agar 2% sodium chloride (TSAN 2) with or without catalase (400 or 600 U) or sodium pyruvate (80 or 160 mg) supplementation. Nucleic acids were extracted from subsamples and subjected to PCR and RT-PCR with hemolysin as the target. Higher recoveries of V. vulnificus were obtained with spread plating compared to pour plating ( P<0.05). The addition of sodium pyruvate (80 mg) or catalase (400 U) significantly increased cell recovery ( P<0.05). PCR amplification signals were stronger than RT-PCR signals at each timepoint, and results were generally consistent between TSAN 2 and ASW for each strain. These results will aid in the design of optimum methods to recover and/or detect V. vulnificus cells subjected to sublethal stress that might be encountered in food processing and storage.

Improved recovery of chlorine-stressed coliforms with pyruvate supplemented media

The addition of sodium pyruvate to standard membrane-filtration media for enumeration of coliforms and E. coli is shown to improve the recovery of chlorine-stressed viable organisms.

Improved recovery of chlorine-stressed coliforms with pyruvate supplemented media

The addition of sodium pyruvate to standard membrane-filtration media for enumeration of coliforms and E. coli is shown to improve the recovery of chlorine-stressed viable organisms.

Peroxidase and tyrosinase are present in secondary lysosomes that degrade photosensory membranes of the crayfish photoreceptor: possible role in pigment granule formation.

Three enzymes (acid phosphatase, peroxidase, and tyrosinase) were localized by electron microscopy within the retina of crayfish Orconectes limosus. Peroxidase activity was observed only in lamellar bodies, which are secondary lysosomes and degrade photosensory membrane. After H2O2 was omitted from the reaction medium, peroxidase activity in lamellar bodies was partly inhibited but was not missing completely. After addition of sodium pyruvate, which inhibits endogenous generation of H2O2, staining of lamellar bodies was absent. Tyrosinase activity was found in lamellar bodies and in small vesicles within the rhabdoms similar to those found positive for acid phosphatase. Granules (500-700 nm in diameter) with an electron opaque matrix and mature screening pigment granules showed tyrosinase activity. Moreover, lamellar structures within membrane-bound organelles that additionally contained screening pigment-like granules were electron dense because of tyrosinase activity. After addition of phenylthiourea (PTU) to the incubation medium, lamellar bodies did not generally contain electron dense deposits, although weak staining of single membranes still was sometimes observed. After addition of sodium pyruvate in combination with PTU, no staining was detected. The possible role of tyrosinase in ommochrome synthesis within secondary lysosomes that degrade photosensory membrane is discussed.

Comparison of three media used for the enumeration of heat-injured Botrytis cinerea

The efficacy of three media (antibiotic-supplemented plate count agar (3 days, 21 °C); antibiotic–pyruvate–supplemented plate count agar (3 days, 21 °C); and trypticase soy agar (16 h, followed by 2.5 days on antibiotic-supplemented plate count agar, 21 °C)) for recovery of heat-injured Botrytis cinerea spores was compared using hydrophobic grid membrane filters. The filters restrict spreading of fungal colonies and facilitate the transfer of fungal spores from pre-enrichment trypticase soy agar to antibiotic-supplemented plate count agar. Counts obtained from pre-enrichment on trypticase soy agar were significantly larger than counts obtained from the other media (α = 0.05). Addition of sodium pyruvate to antibiotic-supplemented plate count agar did not increase spore recovery (α = 0.05). The use of trypticase soy agar as a pre-enrichment medium should be considered for use when antibiotic-supplemented plate count agar is employed in the enumeration of heat-stressed fungi.

Establishment and characterization of a new cell line from a hereditary renal rat tumor.

A new cloned tissue culture cell line, HRRT, has been established from a hereditary renal rat tumor. Electron microscopic studies indicate that the cells are fibroblasts. The cell line has been maintained in monolayer cultures for more than 3 yr and multiplies with a population doubling time of 21 h. The HRRT cells were found to have a plating efficiency of 50% and were not able to grow at low serum concentrations; they did not multiply in suspension culture and were unable to form colonies in soft agar or to proliferate in the aggregate form. The HRRT cells did not form tumors in nude mice. The new cell line was adapted to grow in serum-free medium by decreasing the serum concentration gradually over a period of several months and by addition of sodium pyruvate, insulin, alanine, serine, ribose, thymidine, and uridine to the medium. In serum-free medium HRRT cells multiplied with a population doubling time of 42 h and were not able to form colonies in soft agar.

On the transformation of coliform types, proper and irregular forms

Physiological changes have been induced in strains of E. coli I and its irregular forms, using a basal synthetic medium and addition of sodium pyruvate, or glucose, or yeast extract, or peptone. Changes in biochemical patterns, using the IMViC reactions plus growth at 44.5 degrees C, were induced, thus brining transformations between the regular types and irregular forms. Ultra-violet irradiation of E. coli I for 90 seconds may turn it into a methyl-red negative strain, while education of E. coli III for producing acid and gas at 44.5 degrees C was possible. Thus, E. coli I could be transformed to give the same reactions as the untyped form 1 of Geldreich (1966) through U.V. irradiation, while E. coli III may give the biochemical reactions of E. coli I via heat-resistance and training in MacConkey broth. The relationship between regular and irregular coliforms was also verified by phage typing. The results obtained were discussed both qualitatively, using "lytic activity", and quantitatively in terms of "efficiency of plating", especially with sewage-derived phages and irregular coliforms.

Surface analysis and depth profiles of calcium in hepatoma cells during pyruvate-induced DNA synthesis

Induction of DNA synthesis is associated with increased uptake of calcium in cultured cells. Calcium distribution within the plasma membrane and adjacent cytoplasmic layers of hepatoma cells was investigated with X-ray photoelectron spectroscopy and oxygen plasma etching. Cells in minimal growth medium initiate active DNA synthesis 16 h after addition of sodium pyruvate. Cells stimulated with pyruvate and pyruvate-free controls were analysed by X-ray photoelectron spectroscopy—oxygen plasma etching at 0–40 Å (layer I), 0–450 Å (layer II) and 0–4000 Å (layer III) from the outer cell surface. Calcium concentrations were elevated in induced cells compared with controls: +20% in layer I, +60% in layer II, and +300% in layer III. As the plasma membrane is 90–120 Å thick, these results indicate that pyruvate-induced DNA symthesis is preceded by an increase in calcium, most marked in the cytoplasm subjacent to the plasma membrane, moderate at its inner surface, and minimal at its outer surface.

Cytochemical localization of peroxidatic activity of catalase in rat hepatic microbodies (peroxisomes).

Prominent staining of rat hepatic microbodies was obtained by incubating sections of aldehyde-fixed rat liver in a modified Graham and Karnovsky's medium for ultrastructural demonstration of peroxidase activity. The electron-opaque reaction product was deposited uniformly over the matrix of the microbodies. The microbodies were identified by their size, shape, presence of tubular nucleoids, and other morphologic characteristics, and by their relative numerical counts. The staining reaction was inhibited by the catalase inhibitor, aminotriazole, and by KCN, azide, high concentrations of H(2)O(2), and by boiling of sections. These inhibition studies suggest that the peroxidatic activity of microbody catalase is responsible for the staining reaction. In the absence of exogenous H(2)O(2) appreciable staining of microbodies was noted only after prolonged incubation. Addition of sodium pyruvate, which inhibits endogenous generation of H(2)O(2) by tissue oxidases, or of crystalline catalase, which decomposes such tissue-generated H(2)O(2), completely abolished microbody staining in the absence of H(2)O(2). Neither diaminobenzidine nor the product of its oxidation had any affinity to bind nonenzymatically to microbody catalase and thus stain these organelles. The staining of microbodies was optimal at alkaline pH of 8.5. The biological significance of this alkaline pH in relation to the similar pH optima of several microbody oxidases is discussed. In addition to staining of microbodies, a heat-resistant peroxidase activity is seen in some of the peribiliary dense bodies. The relation of this reaction to the peroxidase activity of lipofuscin pigment granules is discussed.