COMPARATIVE BIOCHEMICAL STUDIES ON NORMAL AND ON POLIOMYELITIS INFECTED TISSUE CULTURES: III. ENZYME ASSAYS ON HOMOGENATES OF SURVIVING NORMAL RHESUS KIDNEY. EFFECT OF SYNTHETIC NUTRIENT MIXTURES

1956 ◽  
Vol 34 (3) ◽  
pp. 619-636 ◽  
Author(s):  
Ernest Kovacs
Keyword(s):  
Synthetic Medium ◽  
Enzyme Assays ◽  
Fresh Tissue ◽  
Normal Tissues ◽  
Salt Mixtures ◽  
Tissue Homogenates ◽  
Synthetic Media ◽  
Tissue Cultivation ◽  
Enzyme Alkaline Phosphatase

Investigation of enzyme systems in normal tissues is the main subject of this communication. The activity of phosphatases, nucleotidases, and nucleases was explored in "brei" prepared with water, physiological salt mixtures, and synthetic media used for tissue cultivation, to compare the influence of these diluents. The depression of acid phosphatase by medium 597, noted previously in cells growing in vitro, was confirmed in fresh tissue and definitely proved on purified enzyme. Alkaline phosphatase was activated by the same synthetic medium. Differences in kinetics were observed when the effect of medium 597 was examined on 5-nucleotidase concentrate of Russel's viper venom and on pentanucleotidase of fresh kidney. Crystalline RNA-se was inhibited, in a manner similar to the nucleases of kidney extracts. Both phosphomonoesterases were enhanced by the less complex nutrient 697. The deterioration of DNA-se in medium 199 upon storage was striking. Significant observations were gathered on enzymes of concentrated and very diluted (1: 1600) tissue homogenates, on extracts, and on isolated enzymes of various grades of purity.

COMPARATIVE BIOCHEMICAL STUDIES ON NORMAL AND ON POLIOMYELITIS INFECTED TISSUE CULTURES: III. ENZYME ASSAYS ON HOMOGENATES OF SURVIVING NORMAL RHESUS KIDNEY. EFFECT OF SYNTHETIC NUTRIENT MIXTURES

1956 ◽  
Vol 34 (1) ◽  
pp. 619-636 ◽  
Author(s):  
Ernest Kovacs
Keyword(s):  
Synthetic Medium ◽  
Enzyme Assays ◽  
Fresh Tissue ◽  
Normal Tissues ◽  
Salt Mixtures ◽  
Tissue Homogenates ◽  
Synthetic Media ◽  
Tissue Cultivation ◽  
Enzyme Alkaline Phosphatase

Investigation of enzyme systems in normal tissues is the main subject of this communication. The activity of phosphatases, nucleotidases, and nucleases was explored in "brei" prepared with water, physiological salt mixtures, and synthetic media used for tissue cultivation, to compare the influence of these diluents. The depression of acid phosphatase by medium 597, noted previously in cells growing in vitro, was confirmed in fresh tissue and definitely proved on purified enzyme. Alkaline phosphatase was activated by the same synthetic medium. Differences in kinetics were observed when the effect of medium 597 was examined on 5-nucleotidase concentrate of Russel's viper venom and on pentanucleotidase of fresh kidney. Crystalline RNA-se was inhibited, in a manner similar to the nucleases of kidney extracts. Both phosphomonoesterases were enhanced by the less complex nutrient 697. The deterioration of DNA-se in medium 199 upon storage was striking. Significant observations were gathered on enzymes of concentrated and very diluted (1: 1600) tissue homogenates, on extracts, and on isolated enzymes of various grades of purity.

scholarly journals Effects of Dietary Anaplerotic and Ketogenic Energy Sources on Renal Fatty Acid Oxidation Induced by Clofibrate in Suckling Neonatal Pigs

2020 ◽  
Vol 21 (3) ◽  
pp. 726
Author(s):  
Xi Lin ◽  
Brandon Pike ◽  
Jinan Zhao ◽  
Yu Fan ◽  
Yongwen Zhu ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Palmitic Acid ◽  
Fatty Acid Oxidation ◽  
Energy Sources ◽  
Acid Oxidation ◽  
Peroxisome Proliferator ◽  
Fresh Tissue ◽  
Peroxisome Proliferator Activated Receptor ◽  
Tissue Homogenates

Maintaining an active fatty acid metabolism is important for renal growth, development, and health. We evaluated the effects of anaplerotic and ketogenic energy sources on fatty acid oxidation during stimulation with clofibrate, a pharmacologic peroxisome proliferator-activated receptor α (PPARα) agonist. Suckling newborn pigs (n = 72) were assigned into 8 dietary treatments following a 2 × 4 factorial design: ± clofibrate (0.35%) and diets containing 5% of either (1) glycerol-succinate (GlySuc), (2) tri-valerate (TriC5), (3) tri-hexanoate (TriC6), or (4) tri-2-methylpentanoate (Tri2MPA). Pigs were housed individually and fed the iso-caloric milk replacer diets for 5 d. Renal fatty acid oxidation was measured in vitro in fresh tissue homogenates using [1-14C]-labeled palmitic acid. The oxidation was 30% greater in pig received clofibrate and 25% greater (p < 0.05) in pigs fed the TriC6 diet compared to those fed diets with GlySuc, TriC5, and Tri2MPA. Addition of carnitine also stimulated the oxidation by twofold (p < 0.05). The effects of TriC6 and carnitine on palmitic acid oxidation were not altered by clofibrate stimulation. However, renal fatty acid composition was altered by clofibrate and Tri2MPA. In conclusion, modification of anaplerosis or ketogenesis via dietary substrates had no influence on in vitro renal palmitic acid oxidation induced by PPARα activation.

scholarly journals Expression Profiling of Virulence and Pathogenicity Genes of Xanthomonas axonopodis pv. citri

2005 ◽  
Vol 187 (3) ◽  
pp. 1201-1205 ◽  
Author(s):  
Gustavo Astua-Monge ◽  
Juliana Freitas-Astua ◽  
Gisele Bacocina ◽  
Juliana Roncoletta ◽  
Sérgio A. Carvalho ◽  
...  
Keyword(s):  
Data Analysis ◽  
Expression Profiling ◽  
Synthetic Medium ◽  
In Vitro System ◽  
Xanthomonas Axonopodis ◽  
Transcriptional Alterations ◽  
Synthetic Media ◽  
Media Data ◽  
Dna Macroarrays

ABSTRACT DNA macroarrays of 279 genes of Xanthomonas axonopodis pv. citri potentially associated with pathogenicity and virulence were used to compare the transcriptional alterations of this bacterium in response to two synthetic media. Data analysis indicated that 31 genes were up-regulated by synthetic medium XVM2, while only 7 genes were repressed. The results suggest that XVM2 could be used as an in vitro system to identify candidate genes involved in pathogenesis of X. axonopodis pv. citri.

scholarly journals LATENT VIRAL INFECTION OF CELLS IN TISSUE CULTURE

1956 ◽  
Vol 103 (6) ◽  
pp. 765-775 ◽  
Author(s):  
Karl M. Johnson ◽  
Herbert R. Morgan
Keyword(s):  
Amino Acids ◽  
Viral Infection ◽  
Synthetic Medium ◽  
Water Soluble ◽  
Virus Growth ◽  
Viral Multiplication ◽  
Minimum Number ◽  
Synthetic Media ◽  
Stimulation Of

When chick embryo tissues cultivated for 13 days in Hanks's balanced salt solution (BSS) were infected with psittacosis virus (6BC), they did not support active viral multiplication until synthetic medium 199 of Parker (3) was added. By testing various combinations of the substances in this and other synthetic media, it was found that the minimum number of compounds required to effectively stimulate virus growth in the presence of BSS comprised the amino acids and water-soluble vitamins found in medium 199. Addition of either amino acids or water-soluble vitamins alone to BSS resulted in only slight stimulation of viral proliferation. Many constituents of the synthetic media were found not to be essential to the stimulation of viral multiplication. The following substances added to a medium containing amino acids and water-soluble vitamins in BSS failed to increase the quantity of virus produced: diphosphopyridine nucleotide (DPN), triphosphopyridine nucleotide (TPN), coenzyme A, the fat-soluble vitamins, ribose sugars, and three biological reducing agents: cysteine, glutathione, and ascorbic acid. Among other substances that proved to be not essential a group of purines and pyrimidines present in medium 199 were found to be probably toxic to cells in the concentrations used, since virus titers were lower in media containing these compounds than in those from which they were absent. A change in the nutritional status of these cells involving amino acids and water-soluble vitamins has thus permitted to transform a latent, undetectable viral infection to an inactive infection in vitro.

THE METABOLISM OF ANIMAL TISSUES CULTIVATED IN VITRO: III. AMINO ACID METABOLISM OF STRAIN L CELLS IN COMPLETELY SYNTHETIC MEDIA

1958 ◽  
Vol 36 (1) ◽  
pp. 771-782 ◽  
Author(s):  
Arthur E. Pasieka ◽  
Helen J. Morton ◽  
Joseph F. Morgan
Keyword(s):  
Amino Acid ◽  
Glutamic Acid ◽  
Culture Medium ◽  
Synthetic Medium ◽  
Amino Acid Content ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
L Cells ◽  
Synthetic Media

Strain L cells, of mouse fibroblastic origin, have been cultivated in vitro in completely synthetic medium M 150 and in various modifications of this medium. The amino acid changes in the nutrient medium during cell cultivation have been studied by paper chromatography. A characteristic pattern of amino acid uptake and accumulation in the medium has been found. No change in the alanine concentration was observed but omission of alanine from the culture medium resulted in its accumulation in appreciable amounts. Omission of glutamic acid did not alter the pattern of amino acid changes by the cells. Omission of glutamine increased the uptake of amino acids and prevented amino acid accumulation. Omission of both glutamic acid and glutamine resulted in a virtual cessation of amino acid changes in the culture medium. Strain L cells decreased the adenine content of the medium and produced small amounts of hypoxanthine. These changes were not affected by alterations in the amino acid content of the medium. Omission of glutamic acid and glutamine from the culture medium did not cause an appreciable decrease in cell population or apparent degeneration of the cultures over a 30-day period.

Effect of homologous follicular fluid from medium-sized and large follicles on in vitro maturation of equine cumulus - oocyte complexes

2005 ◽  
Vol 17 (6) ◽  
pp. 651 ◽  
Author(s):  
Valéria Amorim Conforti ◽  
Dirk K. Vanderwall ◽  
Gordon L. Woods
Keyword(s):  
Follicular Fluid ◽  
In Vitro Maturation ◽  
Transvaginal Ultrasound ◽  
Synthetic Medium ◽  
Control Medium ◽  
Ultrasound Guided ◽  
Suitable Alternative ◽  
Follicle Aspiration ◽  
Synthetic Media

The in vitro maturation (IVM) of equine oocytes is typically performed using various synthetic media; however, an optimal IVM system for equine oocytes has not been developed. The aim of the present study was to evaluate the effects of two types of follicular fluid (FF) obtained from cyclic mares and two incubation intervals for the IVM of equine cumulus–oocyte complexes (COCs). Follicular fluid was collected from medium-sized (20–29 mm diameter) and large (≥30 mm; post-human chorionic gonadotrophin administration) follicles using transvaginal ultrasound-guided follicle aspiration. Compact (n = 232) and non-compact (n = 183) COCs obtained from a slaughterhouse were incubated separately in the following groups: (1) FF from medium follicles for 24 h; (2) FF from large follicles for 24 h; (3) control (synthetic) medium for 24 h; (4) FF from medium follicles for 24 h then FF from large follicles for an additional 24 h; (5) FF from large follicles for 48 h; and (6) control medium for 48 h. For compact COCs, there was a tendency (P = 0.06) for more COCs incubated in FF from large follicles for 24 h to reach metaphase II compared with those incubated in control medium for 24 h (58% v. 35%, respectively). More (P < 0.05) compact COCs had degenerated after incubation in control medium for 48 h compared with all other groups (51% v. 14–24%, respectively). For non-compact COCs, incubation in FF from medium follicles for 24 h resulted in more (P = 0.05) COCs at metaphase II compared with control medium for 48 h (58% v. 29%, respectively). These results indicate that homologous FF from cyclic mares is a suitable alternative for the IVM of equine COCs and that it may be superior to conventional media for longer (i.e. >24 h) incubation intervals.

scholarly journals Platelet storage for transfusion in synthetic media: further optimization of ingredients and definition of their roles

Blood ◽  
1995 ◽  
Vol 86 (10) ◽  
pp. 3951-3960 ◽  
Author(s):  
S Murphy ◽  
T Shimizu ◽  
J Miripol
Keyword(s):  
Lactic Acid ◽  
Acid Production ◽  
Lactic Acid Production ◽  
Synthetic Medium ◽  
Lactate Production ◽  
Hydrogen Ion ◽  
In Vitro Tests ◽  
Autologous Plasma ◽  
Synthetic Media

Currently, most platelet concentrates (PC) are stored for transfusion at 20 degrees C to 24 degrees C in autologous plasma. There are potential advantages in replacing some of this plasma with a synthetic medium. In this study, our major goals were to define the optimal ingredients and their concentrations in such a medium and to gain insight into the mechanism by which each ingredient confers benefit. In addition, we wished to validate a new polyvinyl chloride container plasticized with di-n-decyl phthalate (DnDP) for PC storage. PC derived from donations of whole blood were stored for 7 days in autologous plasma or a basic synthetic medium (BSM) containing 15 mmol/L glucose, 21 mmol/L citrate, and physiologic concentrations of salts other than bicarbonate within either the DnDP container or a licensed polyolefin container, PL-732. Metabolic events were characterized and a panel of in vitro tests were used to monitor platelet quality as systematic changes in the BSM were made. Platelet quality was as least as good, if not better, after storage in DnDP in comparison to PL-732. pH consistently decreased to less than 6.0 because of inadequate buffering of lactic acid in BSM alone. However, pH and the in vitro tests were well maintained by either the serial addition of bicarbonate (BSM + B) or the addition of at least 15 mmol/L acetate and 10 mmol/L phosphate (BSM + AP). The benefits of BSM + AP were traced to a decrease in lactic acid production by 33% and 19% relative to plasma and BSM + B, respectively, and the vigorous oxidation of acetate (0.66 +/- 0.09 mmol/d/10(12 platelets). The rates of lactate production and acetate consumption were similar and the pH during storage correlated with difference between the two rates, suggesting that acetate oxidation has an alkalinizing effect equivalent on a molar basis to the acidifying effect of production of lactate and a hydrogen ion. When pyruvate replaced acetate, it was also metabolized vigorously (0.52 +/- 0.06 mmol/d/10(12) platelets). Its presence suppressed lactic acid production by 44% relative to BSM + B and allowed maintenance of pH and platelet quality similar to what is achieved with acetate. The results strongly suggest that the benefit from acetate (or pyruvate) is derived from its oxidation and the use of a hydrogen ion during that oxidation. For reasons that are not yet clear, the omission of phosphate resulted in pH decrease to less than 6.0 in 3 of 9 PC even with acetate present.(ABSTRACT TRUNCATED AT 400 WORDS)

THE METABOLISM OF ANIMAL TISSUES CULTIVATED IN VITRO: III. AMINO ACID METABOLISM OF STRAIN L CELLS IN COMPLETELY SYNTHETIC MEDIA

1958 ◽  
Vol 36 (7) ◽  
pp. 771-782 ◽  
Author(s):  
Arthur E. Pasieka ◽  
Helen J. Morton ◽  
Joseph F. Morgan
Keyword(s):  
Amino Acid ◽  
Glutamic Acid ◽  
Culture Medium ◽  
Synthetic Medium ◽  
Amino Acid Content ◽  
Amino Acid Uptake ◽  
Acid Uptake ◽  
L Cells ◽  
Synthetic Media

Strain L cells, of mouse fibroblastic origin, have been cultivated in vitro in completely synthetic medium M 150 and in various modifications of this medium. The amino acid changes in the nutrient medium during cell cultivation have been studied by paper chromatography. A characteristic pattern of amino acid uptake and accumulation in the medium has been found. No change in the alanine concentration was observed but omission of alanine from the culture medium resulted in its accumulation in appreciable amounts. Omission of glutamic acid did not alter the pattern of amino acid changes by the cells. Omission of glutamine increased the uptake of amino acids and prevented amino acid accumulation. Omission of both glutamic acid and glutamine resulted in a virtual cessation of amino acid changes in the culture medium. Strain L cells decreased the adenine content of the medium and produced small amounts of hypoxanthine. These changes were not affected by alterations in the amino acid content of the medium. Omission of glutamic acid and glutamine from the culture medium did not cause an appreciable decrease in cell population or apparent degeneration of the cultures over a 30-day period.

scholarly journals Activation of MAT2A-RIP1 signaling axis reprograms monocytes in gastric cancer

2021 ◽  
Vol 9 (2) ◽  
pp. e001364
Author(s):  
Yan Zhang ◽  
Hui Yang ◽  
Jun Zhao ◽  
Ping Wan ◽  
Ye Hu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Cell Metabolism ◽  
Vital Role ◽  
Methionine Metabolism ◽  
Promoter Regions ◽  
Normal Tissues ◽  
Methionine Cycle

BackgroundThe activation of tumor-associated macrophages (TAMs) facilitates the progression of gastric cancer (GC). Cell metabolism reprogramming has been shown to play a vital role in the polarization of TAMs. However, the role of methionine metabolism in function of TAMs remains to be explored.MethodsMonocytes/macrophages were isolated from peripheral blood, tumor tissues or normal tissues from healthy donors or patients with GC. The role of methionine metabolism in the activation of TAMs was evaluated with both in vivo analyses and in vitro experiments. Pharmacological inhibition of the methionine cycle and modulation of key metabolic genes was employed, where molecular and biological analyses were performed.ResultsTAMs have increased methionine cycle activity that are mainly attributed to elevated methionine adenosyltransferase II alpha (MAT2A) levels. MAT2A modulates the activation and maintenance of the phenotype of TAMs and mediates the upregulation of RIP1 by increasing the histone H3K4 methylation (H3K4me3) at its promoter regions.ConclusionsOur data cast light on a novel mechanism by which methionine metabolism regulates the anti-inflammatory functions of monocytes in GC. MAT2A might be a potential therapeutic target for cancer cells as well as TAMs in GC.

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