The Glucose, Insulin and Glutamine Requirements of Suspension Cultures of HeLa Cells in a Difined Culture Medium

1971 ◽  
Vol 9 (2) ◽  
pp. 529-537
Author(s):  
G. J. BLAKER ◽  
J. R. BIRCH ◽  
S. J. PIRT
Keyword(s):  
Amino Acid ◽  
Hela Cells ◽  
Growth Yield ◽  
Dry Weight ◽  
Defined Medium ◽  
Cell Populations ◽  
Serum Supplement ◽  
Protamine Sulphate ◽  
Protamine Zinc Insulin ◽  
Maximum Cell

The serum supplement in a defined medium for the growth of HeLa cells could be replaced by protamine-zinc-insulin (0.2 u./ml). Insulin (0.4 u./ml) replaced the growth-stimulatory properties of protamine-zinc-insulin, whilst protamine sulphate (5 µg/ml) was found to be toxic to the cells. The addition of insulin to cultures depleted of insulin increased both cell growth rates and maximum cell populations. In the defined medium, HeLa cells could only utilize glutamate when a small amount of glutamine was included. Glucose, at a level of 2 mg/ml, was shown to limit maximum cell populations. The growth yield from glucose was 295 µg cell dry weight/mg glucose. When the medium glucose concentration was increased to 4 mg/ml, HeLa cell populations in excess of 16 x 105 cells (i.e. 640 µg dry weight)/ml were routinely achieved in the defined medium supplemented with insulin. Growth is then limited by the amino acid supply. Increasing the amino acid concentration of the medium by 50% raised the maximum cell population to 23.5x105 cells (i.e. 940 µg dry weight)/ml.

The Quantitative Glucose and Mineral Nutrient Requirements of Mouse LS (Suspension) Cells in Chemically Defined Medium

1971 ◽  
Vol 8 (3) ◽  
pp. 693-700
Author(s):  
J. R. BIRCH ◽  
S. J. PIRT
Keyword(s):  
Growth Response ◽  
Potassium Concentration ◽  
Growth Yield ◽  
Dry Weight ◽  
Defined Medium ◽  
Mineral Nutrient ◽  
Nutrient Requirements ◽  
Suspension Cells ◽  
Glucose Phosphate ◽  
Maximum Population Density

The influences of glucose, phosphate, magnesium and potassium concentration on growth rate and maximum population density were studied. From the data obtained the growth yields for glucose and phosphorus were found to be 2.4 x 105 cells (i.e. 0.76 mg dry weight)/mg glucose and 2.2 x 105 (i.e. 69 µg dry weight)/µg P. The growth response to magnesium and potassium was complex since there were threshold concentrations of these metals below which no growth occurred. The growth yield for potassium (2.4 x 105 cells (i.e. 75 µg dry weight)/µg K) was obtained by measuring potassium uptake during growth. Omission of calcium ions from the medium resulted in poor reproducibility of growth.

The quantitative requirements of human diploid cells (strain MRC-5) for amino acids, vitamins and serum

1975 ◽  
Vol 17 (3) ◽  
pp. 397-411
Author(s):  
K. Lambert ◽  
S.J. Pirt
Keyword(s):  
Amino Acids ◽  
Growth Factor ◽  
Inorganic Compounds ◽  
Calf Serum ◽  
Essential Amino Acids ◽  
Defined Medium ◽  
Cell Populations ◽  
Maximum Cell ◽  
Human Diploid Cells ◽  
Diploid Cells

The uptakes of all essential amino acids, vitamins (except riboflavin), glucose and serum during growth of human diploid cells (MRC-5) were determined. The amino acid uptakes varied considerably with the conditions of culture. The glucose requirement is several times greater than that for mouse LS or human HeLa cells. These analytical results were used to modify the medium so as to ensure that an excess of all defined medium constituents was present and pH was not limiting during study of the serum requirements. It was then found that maximum cell populations were directly proportional to the serum concentration. Hence the growth was limited by the supply of an unknown growth factor in serum. The serum growth factor was not replaced by a mixture of over 60 vitamins, co-enzymes, hormones and other organic and inorganic compounds considered to be possible growth factors, although this mixture did not lower the growth rate and somewhat (22%) increased the yield from the serum growth factor. The unit of serum growth factor is precisely defined in terms of the amount in a standard batch of calf serum. This standard contains 10 units/ml whereas the other batch of serum used contained only 5 units/ml.

Minimization of activated sludge production by chemically stimulated energy spilling

2000 ◽  
Vol 42 (12) ◽  
pp. 189-200 ◽  
Author(s):  
G.-H. Chen ◽  
H.-K. Mo ◽  
S. Saby ◽  
W.-k. Yip ◽  
Y. Liu
Keyword(s):  
Activated Sludge ◽  
Growth Yield ◽  
Dry Weight ◽  
Staining Technique ◽  
Microbial Culture ◽  
Dapi Staining ◽  
E Coli ◽  
Energy Spilling ◽  
Activated Sludge Processes ◽  
Sludge Production

Minimization of excess sludge production in activated sludge processes has been pursued around the world in order to meet stringent environmental regulations on sludge treatment and disposal. To achieve this goal, physical, chemical, and biological approaches have been proposed. In this paper, a chemical compound, 3,3′,4′,5-tetrachlorosalicylanilide (TCS) was tested for enhancing microbial energy spilling of the sludgeso as to minimize its growth. In order to examine this, an exploratory study was conducted using both batch and continuous activated sludge cultures. Batch experiments with these two cultures were carried out at different initial concentrations of TCS. It has been confirmed that an addition of TCS is effective in reducing the production of both the sludge cultures, particularly the continuous culture where the observed growth yield was reduced by around 70%, when the initial TCS concentration was 0.8 ppm. Meanwhile, the substrate removal activity of this culture was found not to be affected at this TCS concentration. To further evaluate the TCS effect, a pure microbial culture of E. coli was employed. Batch experiment results with this culture implied that TCS might be able to reduce the cell density of E. coli drastically when an initial TCS concentration was greater than 0.12 ppm. It was also found that TCS was not toxic to this type of bacteria. Microscopic examinations with a 4′, 6-diamidino-2-phenylindole (DAPI) staining technique revealed that TCS neither affected the cell division nor altered the cell size of E. coli. However, both the cell ATP content and the cell dry weight were reduced significantly with the addition of TCS.

scholarly journals Enterovirus 70 Receptor Utilization Is Controlled by Capsid Residues That Also Regulate Host Range and Cytopathogenicity

2007 ◽  
Vol 81 (16) ◽  
pp. 8648-8655 ◽  
Author(s):  
Melissa Stewart Kim ◽  
Vincent R. Racaniello
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Rhesus Monkey ◽  
Host Range ◽  
Hela Cells ◽  
Lytic Replication ◽  
Monkey Kidney ◽  
Kidney Cells ◽  
Cytopathic Effects ◽  
Axis Of Symmetry

ABSTRACT Enterovirus type 70, an etiologic agent of acute hemorrhagic conjunctivitis, may bind different cellular receptors depending on cell type. To understand how EV70-receptor interaction is controlled, we studied two variants of the virus with distinct receptor utilization. EV70-Rmk, derived by passage in rhesus monkey kidney cells, replicates poorly in HeLa cells and does not cause cytopathic effects. Decay accelerating factor (DAF) is not a cell receptor for EV70-Rmk. Passage of EV70-Rmk in HeLa cells lead to isolation of EV70-Dne, which does not replicate in rhesus monkey kidney cells but grows to high titers in HeLa cells and causes cytopathic effects. DAF is sufficient for cell entry of EV70-Dne. EV70-Rmk replicates in human eye and brain-derived cell lines, whereas the Dne strain replicates only in HeLa cells and in conjunctiva-derived 15C4 cells. The two EV70 strains differ by five amino acid changes in the viral capsid. Single substitution of four of the five EV70-Rmk amino acids with the residue from EV70-Dne leads to lytic replication in HeLa cells. Conversely, substitution of any of the five EV70-Dne amino acids with the EV70-Rmk amino acid does not alter replication in HeLa cells. Three of these capsid amino acids are predicted to be located in the canyon encircling the fivefold axis of symmetry, one amino acid is found at the fivefold axis of symmetry, and one is located the interior of the capsid. The five EV70 residues define a region of the capsid that controls viral host range, DAF utilization, and cytopathogenicity.

The Evidence for Reduced Glyphosate Efficacy on Acetolactate Synthase–Inhibiting Herbicide-Resistant Yellow Nutsedge (Cyperus esculentus)

Weed Science ◽  
2016 ◽  
Vol 64 (3) ◽  
pp. 389-398
Author(s):  
Parsa Tehranchian ◽  
Jason K. Norsworthy ◽  
Matheus Palhano ◽  
Nicholas E. Korres ◽  
Scott McElroy ◽  
...  
Keyword(s):  
Amino Acid ◽  
Production Systems ◽  
Acetolactate Synthase ◽  
Dry Weight ◽  
Cross Resistance ◽  
Purple Nutsedge ◽  
Yellow Nutsedge ◽  
Epsps Gene ◽  
Treated Leaf ◽  
Als Inhibitors

A yellow nutsedge biotype (Res) from an Arkansas rice field has evolved resistance to acetolactate synthase (ALS)-inhibiting herbicides. TheResbiotype previously exhibited cross-resistance to ALS inhibitors from four chemical families (imidazolinone, pyrimidinyl benzoate, sulfonylurea, and triazolopyrimidine). Experiments were conducted to evaluate alternative herbicides (i.e., glyphosate, bentazon, propanil, quinclorac, and 2,4-D) currently labeled in Arkansas rice–soybean production systems. Based on the percentage of aboveground dry weight reduction, control of the yellow nutsedge biotypes with the labeled rate of bentazon, propanil, quinclorac, and 2,4-D was < 44%. Glyphosate (867 g ae ha−1) resulted in 68 and > 94% control of theResand susceptible yellow nutsedge biotypes, respectively, at 28 d after treatment. Dose-response studies were conducted to estimate the efficacy of glyphosate on theResbiotype, three susceptible yellow nutsedge biotypes, and purple nutsedge. Based on the dry weights, theResbiotype was ≥ 5- and ≥ 1.3-fold less responsive to glyphosate compared to the susceptible biotypes and purple nutsedge, respectively. Differences in absorption and translocation of radiolabeled glyphosate were observed among the yellow nutsedge biotypes and purple nutsedge. The susceptible biotype had less14C-glyphosate radioactivity in the tissues above the treated leaf and greater radioactivity in tissues below the treated leaf compared to theResbiotype and purple nutsedge. Reduced translocation of glyphosate in tissues below the treated leaf of theResbiotype could be a reason for the lower glyphosate efficacy in theResbiotype. No amino acid substitution that would correspond to glyphosate resistance was found in the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene of theResbiotype. However, an amino acid (serine) addition was detected in the EPSPS gene of theResbiotype; albeit, it is not believed that this addition contributes to lower efficacy of glyphosate in this biotype.

scholarly journals Sulfur application alleviates chromium stress in maize and wheat

2020 ◽  
Vol 18 (1) ◽  
pp. 1093-1104
Author(s):  
Grzegorz Kulczycki ◽  
Elżbieta Sacała
Keyword(s):  
Adverse Effects ◽  
Mineral Nutrition ◽  
Elemental Sulfur ◽  
Marked Decrease ◽  
Total Yield ◽  
Growth Yield ◽  
Dry Weight ◽  
Total Biomass ◽  
Cr Content ◽  
Chromium Stress

AbstractThis study aimed to examine the influence of increasing doses of chromium (Cr) (26, 39, and 52 mg kg−1 soil) and elemental sulfur (S) (60 mg kg−1 soil) on growth, yield, and mineral nutrition in wheat and maize. Macro- and micronutrients and Cr concentrations were determined in the aboveground parts of plants. All examined doses of Cr caused a marked decrease in the fresh and dry weight of maize. Wheat was more tolerant than maize, and lower Cr doses caused a small but statistically significant increase in the total yield. Wheat accumulated more than twofold Cr than maize, and the concentrations increased with higher Cr concentrations in the soil. The application of S significantly improved the total biomass production and lowered the Cr content in both plants. Cr changed the mineral nutrition in both cereals, but the pattern of changes observed was not the same. Applying S alleviated some adverse effects caused by the Cr. Hence, it is concluded that the application of elemental S may be an effective strategy to reduce adverse effects in plants grown on soil contaminated by heavy metals, especially Cr.

Growth of human diploid cells (strain MRC-5) in defined medium; replacement of serum by a fraction of serum ultrafiltrate

1979 ◽  
Vol 35 (1) ◽  
pp. 381-392
Author(s):  
K. Lambert ◽  
S.J. Pirt
Keyword(s):  
Amino Acids ◽  
Active Material ◽  
Maximum Yield ◽  
Calf Serum ◽  
Defined Medium ◽  
Cell Yield ◽  
Maximum Cell ◽  
Macromolecular Component ◽  
Human Diploid Cells ◽  
Diploid Cells

A calf serum ultrafiltrate fraction permitted growth for at least 3.5 generations, including one subculture, of MRC-5 cells in defined medium in the absence of whole serum. The active material has a molecular weight of 10 000 Daltons or less. This suggests that there may be no requirement for a large macromolecular component of serum. The ultrafiltrate was assayed by maximum cell yield from a serum-limited inoculum in a defined medium containing non-limiting amounts of vitamins, amino acids, glucose, a 68-component supplement, iron and methylcellulose. The levels of vitamins, amino acids and glucose were based on quantitative measurements of uptake and the levels of the other components by minimum amount required for maximum yield in defined medium without ultrafiltrate or serum. With excess ultrafiltrate maximum cell yield was limited by the defined part of the medium, probably the supplement. The cell doubling time in defined medium with ultrafiltrate fractions was 70 h compared with 27 h in the medium with serum. Excess ultrafiltrate did not inhibit growth. The lowered growth rate is attributed to a nutritional deficiency in the supplement.

scholarly journals Effect of arsenic on Amaranthus gangeticus

2018 ◽  
Vol 53 (4) ◽  
pp. 259-264
Author(s):  
MZ Hossain ◽  
Sushmita Dey ◽  
MS Islam
Keyword(s):  
Irrigation Water ◽  
Crop Production ◽  
Growth Yield ◽  
Dry Weight ◽  
Arsenic Contamination ◽  
Pot Experiment ◽  
Production Potential ◽  
Groundwater Arsenic ◽  
Major Nutrients

Groundwater arsenic contamination has become a threat to the crop production potential in the soils of vast areas of Bangladesh. Situation is grave in some districts of the country, particularly the southern part. A pot experiment was conducted to investigate the effects of arsenic treated irrigation water (0, 1, 2, 5 and 10 mgL-1), where a total of ten (10) irrigations were provided thus the treatments received 0, 10, 20, 50, and 100 mg arsenic (As) pot-1. Effects of applied levels of arsenic on Amaranthus gangeticus (Lal shak) were evaluated in terms of the growth, yield, major nutrients’ content, and their translocation in the plant. As treatments significantly reduced (p≤0.05) the dry weight of shoot and root by 19.31% and 44.03% respectively. Both total and available concentrations of nitrogen (N), potassium (K) and sulfur (S) were significantly (p≤ 0.05) suppressed by the As treatments, while only higher three doses significantly (p≤ 0.05) affected both levels of concentrations of phosphorus (P), calcium (Ca) and magnesium (Mg). Translocation coefficients for soil to root for P, K, S, and Mg were significantly reduced (p≤ 0.05), while translocation coefficients for root to shoot were significantly increased (p≤ 0.05) for K and S by 5 and 10 mgL-1 of arsenic treatments.Bangladesh J. Sci. Ind. Res.53(4), 259-264, 2018

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