Influence of S-adenosylmethionine on DAPI-induced fluorescence of polyphosphate in the yeast vacuole

1980 ◽  
Vol 26 (8) ◽  
pp. 912-920 ◽  
Author(s):  
R. A. Allan ◽  
J. J. Miller
Keyword(s):  
Amino Acids ◽  
Brownian Motion ◽  
Aspartic Acid ◽  
Growth Medium ◽  
In Vitro Studies ◽  
Phosphate Solution ◽  
Yeast Vacuole ◽  
Glucose Phosphate ◽  
Ultraviolet Microscopy

Use of the fluorochrome 4′,6-diamidino-2-phenylindole∙2 HCl (DAPI) in ultraviolet microscopy revealed fluorescent objects in Brownian motion within the vacuoles of seven species of yeast. The abundance of these bodies increased when cells of Saccharomyees cerevisiae were transferred from growth medium to a glucose–phosphate solution, indicating that they contain polyphosphate. In addition, the effect on vacuolar fluorescence of supplementing a defined growth medium with amino acids provided evidence that they also contained S-adenosylmethionine. These deductions were supported by in vitro studies of the interaction and fluorescence of polyphosphate, S-adenosylmethionine, and DAPI. Vacuolar fluorescence of cells in suspension in glucose–phosphate solution was less after addition of exogenous arginine, lysine, or glutamine but not after addition of alanine, aspartic acid, or methionine.Mithramycin was superior to DAPI as a fluorochrome for ultraviolet demonstration of yeast nuclei since it stained the nuclei much more intensely and did not fluoresce with other material in the cells.

scholarly journals MICROSPECTROPHOTOMETRY OF NEWLY SYNTHESIZED STARCH IN SITU

1970 ◽  
Vol 18 (6) ◽  
pp. 439-449
Author(s):  
GEORGE E. WHEELER
Keyword(s):  
Absorption Maximum ◽  
In Vitro Studies ◽  
Phosphate Solution ◽  
Starch Digestion ◽  
Spectral Curves ◽  
Similarities And Differences

Many of the cells in stem sections of several Commelinaceae species synthesized much new starch when incubated in buffered 1% glucose 1-phosphate solution. The starch appeared in the cytoplasm rather than in the plastids. Although the starch I2-KI color was uniform within any one cell, there was considerable variation from cell to cell, even in the same section. The colors with I2-KI ranged from blue, through purples to magenta and mahogany. Tests with α-amylase and with β-amylase showed the starch to be amylose. Microspectrophotometrically determined extinction curves, based on the new starch in situ, varied with the visualized color. As expected, starch which stained blue with I2-KI had an absorption maximum in the orange-red wavelengths above 600 mµ; increasingly red I2-KI colors were characterized by shifts of the absorption maximum further into the shorter wavelengths. The course of new starch digestion by α-amylase and by β-amylase was followed visually and with the microspectrophotometer. Similarities and differences between these spectral curves and those published for in vitro studies are pointed out. The difficulties met with in using the microspectrophotometric method are discussed.

scholarly journals Enhancement of aromatase activity by D-aspartic acid in the ovary of the lizard Podarcis s. sicula

Reproduction ◽  
2001 ◽  
pp. 803-808 ◽  
Author(s):  
L Assisi ◽  
V Botte ◽  
A D'Aniello ◽  
MM Di Fiore
Keyword(s):  
Amino Acids ◽  
Aspartic Acid ◽  
Reproductive Cycle ◽  
Catalytic Properties ◽  
Ovarian Tissue ◽  
Experimental Treatment ◽  
Tissue Homogenate ◽  
Aromatase Activity

The present study investigated the role of D-aspartic acid (D-Asp) in ovarian steroidogenesis and its effect on aromatase activity in the lizard, Podarcis s. sicula. It was determined that D-Asp concentrations vary significantly during phases of the reproductive cycle: they vary inversely with testosterone concentrations and directly with oestradiol concentrations in the ovary and plasma. Experimental treatment showed that administration of D-Asp induces a decrease in testosterone and an increase in oestradiol, and that treatment with other amino acids (L-Asp, D-Glu and D-Ala) instead of D-Asp has no effects. Experiments in vitro confirmed these results. Furthermore, these experiments showed an increase in aromatase activity, as the addition of D-Asp either to fresh ovarian tissue homogenate or to acetonic powder of ovarian follicles induced a significant increase in the conversion of testosterone to oestradiol. Aromatase activity is four times greater in the presence of D-Asp than in its absence. However, almost equivalent values of the two K(m) values (both approximately 25 nmol l(-1)) indicate that aromatase has the same catalytic properties in both cases.

Effect of Amygdaloid Kindling on the Content and Release of Amino Acids from the Amygdaloid Complex: In Vivo and In Vitro Studies

2002 ◽  
Vol 65 (3) ◽  
pp. 1240-1249 ◽  
Author(s):  
S. Kaura ◽  
H. F. Bradford ◽  
A. M. J. Young ◽  
M. J. Croucher ◽  
P. D. Hughes
Keyword(s):  
Amino Acids ◽  
Amygdaloid Complex ◽  
In Vitro Studies

The oxygen uptake of Trypanosoma lewisi and Trypanosoma equiperdum, with especial reference to oxygen consumption in the presence of amino-acids

Parasitology ◽  
1958 ◽  
Vol 48 (1-2) ◽  
pp. 149-164 ◽  
Author(s):  
June P. Thurston
Keyword(s):  
Amino Acids ◽  
Oxygen Consumption ◽  
Oxygen Uptake ◽  
Glutamic Acid ◽  
Aspartic Acid ◽  
Yeast Extract ◽  
Casein Hydrolysate ◽  
Phosphate Solution ◽  
Trypanosoma Lewisi ◽  
Rate Of Oxygen Consumption

1. Standard conditions are described for preparing suspensions of washed Trypanosoma lewisi and T. equiperdum in modified Ringer–phosphate solution.2. Oxygen consumption was measured with differential manometers, using microflasks containing 2–5 × 107 trypanosomes in 0·9 ml. of reaction mixture. Measurements of oxygen uptake were carried out at 37° C.3. T. lewisi respired slowly in the absence of substrate for up to 2 hr. The trypanosomes suffered little damage when stored at 5° C. for 24 hr. without substrate. No oxygen uptake was observed with T. equiperdum in the absence of substrate. The trypanosomes were viable after 24 hr. at 5° C. with glucose or glycerol as substrate, but not in the absence of substrate.4. With glucose as substrate, the rate of oxygen consumption by T. lewisi increased with the age of infection. This change was more marked with glutamine as substrate.5. With glucosamine as substrate, the oxygen uptake of T. lewisi was at a slightly lower rate than with glucose. The rate of oxygen uptake was still lower with Na l-glutamic acid, asparagine, aspartic acid, casein hydrolysate, yeast extract and Difco Bacto-peptone. Thirteen other amino-acids had no effect on the motility of the trypanosomes.6. With glycerol as substrate, the oxygen uptake of T. equiperdum was at a slightly lower rate than with glucose. The rate of oxygen uptake was very low with yeast extract, casein hydrolysate and Difco Bacto-peptone. No oxygen uptake or motility was recorded with glutamine, Na l-glutamic acid, glucosamine, asparagine, aspartic acid, dl-alanine, or Na acetate. Thirteen other amino-acids had no effect on the motility of the trypanosomes.7. Ammonia was liberated from glutamine by adult and reproductive phase T. lewisi.

Regulation of Glyconeogenesis from Amino Acids by Acetate in Tetrahymena pyriformis

1973 ◽  
Vol 51 (4) ◽  
pp. 323-331 ◽  
Author(s):  
C. Mavrides
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Glutamic Acid ◽  
Glutamate Dehydrogenase ◽  
Aspartic Acid ◽  
Growth Medium ◽  
Aspartate Aminotransferase ◽  
Alanine Aminotransferase ◽  
Sodium Acetate ◽  
Tetrahymena Pyriformis

The regulation of glyconeogenesis from amino acids by acetate was studied in Tetrahymena pyriformis. Alanine aminotransferase and glutamate dehydrogenase were repressed by 0.1% sodium acetate in the growth medium. Incorporations into the glycogen of washed cells from the respective isotopically labelled amino acids were similarly suppressed.Incorporations into glycogen from uniformly 14C-labelled L-serine, L-leucine, L-isoleucine, L-tyrosine, and DL-β-14C-tyrosine were also suppressed by prior growth in a medium supplemented with 0.1% or 0.3% acetate. Percentage incorporation into glycogen was highest from tyrosine, followed by leucine, isoleucine, and alanine, and lowest from glutamic acid and serine.Supplementation of the medium with 0.25% glucose resulted in repression of the above two enzymes and suppression of incorporation into glycogen from amino acids.Incorporation of aspartic acid into glycogen was negligible and was variously and minimally affected by growth in glucose- or acetate-supplemented media. Aspartate aminotransferase was affected in a like manner.Glycogen content was not significantly affected by growth in media supplemented with 0.1% or 0.3% acetate. On the whole, the data suggest that acetate spares amino acids for glyconeogenesis by a mechanism which entails repression of amino-acid-catabolizing enzymes.

scholarly journals Differences in huperzine A yield when different amino acids were added for in vitro thallus culture of Huperzia serrata (Thunb)Trev and gene expression analysis

2020 ◽  
Author(s):  
Ye YuXuan ◽  
Tu YiSheng ◽  
Yu Xiao ◽  
Huang Qian ◽  
Yuan Huihui
Keyword(s):  
Gene Expression ◽  
Amino Acids ◽  
Aspartic Acid ◽  
Quantitative Pcr ◽  
Dry Weight ◽  
Huperzine A ◽  
Differential Analysis ◽  
Fluorescence Quantitative Pcr ◽  
Alignment Analysis

Abstract Background：the secondary metabolite of H. serrata, huperzine A (HupA) can be used in the treatment of Alzheimer’s disease and can improve the cognitive function of patients. The use of in vitro culture and secondary metabolism engineering to obtain secondary metabolites is the most effective method to solve a lack of HupA sources and protect H. serrata as a natural resource. This study was based on the in vitro thallus culture conditions for H. serrate, and different concentrations of alkaloid precursor amino acids (lysine, aspartic acid, and trytophan) were added. We found that addition of different amino acids to thallus cultures had different effects on HupA accumulation. Transcriptome sequencing was carried out on thalli with significant differences in the HupA content due to treatment with different amino acids for differential analysis, and real-time fluorescence quantitative PCR was used for validation to examine the functional genes involved in exogenous amino acid regulation of HupA accumulation in thalli.Results：We found that addition of 1 mmol·L−1 aspartic acid (D) solution promoted HupA accumulation, at a level of 84.05 μg·g−1 dry weight (DW), which was 1.29-fold that of the control (CK: 65.15 μg·g−1 DW). Addition of 4 mmol·L−1 lysine (K) solution significantly inhibited HupA accumulation, at a level of 48.42 μg·g−1 DW, which was 0.75-fold that of the control.Transcriptome sequencing-bioinformatics alignment analysis of the aforementioned materials showed that in GO alignment analysis, functions were annotated for 16,258 unigenes. From the statistical analysis of the DEGs of the three groups, we found that there were 1046, 782, and 1586 DEGs for CK vs D, CK vs K, and D vs K, respectively, with D vs K having the most DEGs. DEGs that were enriched in KEGG metabolic pathways and validated by fluorescence quantitative PCR included PANK1, GDH2, APX, HA1, ND4L, and COX1. Conclusions：The above results showed that HupA content differences in D and K treatments were directly proportional to DEGs. Gene expression differences are the molecular basis that affects HupA accumulation in in vitro thallus cultures. PANK1 and GDH2 encode enzymes that synthesize an alkaloid intermediate.

In vitro experiments concerning the state of polyphosphate in the yeast vacuole

1984 ◽  
Vol 30 (2) ◽  
pp. 236-246 ◽  
Author(s):  
J. J. Miller
Keyword(s):  
Amino Acids ◽  
Ribonucleic Acid ◽  
Dextran Sulfate ◽  
Insoluble Fraction ◽  
Yeast Cells ◽  
In Vitro Experiments ◽  
Yeast Vacuole ◽  
Low Concentrations ◽  
Cationic Amino Acids

Globules resembling the cytological entity volutin were produced in vitro by mixing solutions of polyphosphate and certain cationic substances known to occur abundantly in the yeast vacuole, i.e., S-adenosylmethionine, Mg2+, and K+. Other cationic substances found to form globules with polyphosphate were Mn2+, Zn2+, Ca2+, Cd2+, S-adenosylethionine, spermidine, and spermine. Cationic amino acids did not form a precipitate with polyphosphate and at low concentrations they inhibited precipitation of globules of polyphosphate–S-adenosylmethionine. Two other polyanions, dextran sulfate and ribonucleic acid, also formed globular precipitates with S-adenosylmethionine. Conditions affecting the formation and stability of polyphosphate–S-adenosylmethionine precipitates were studied in an attempt to reproduce certain characteristics of vacuolar polyphosphate "granules." Procedures commonly used for extraction of polyphosphate from yeast were applied to in vitro granules and the results indicated that part of the "acid-insoluble" fraction of the polyphosphate extracted from yeast cells may exist in combination with S-adenosylmethionine.

In-vitro studies on intestinal malabsorption of amino-acids in young chicks infected with Eimeria acervulina

1975 ◽  
Vol 4 (1) ◽  
pp. 11-16
Author(s):  
D. S. P. Patterson ◽  
L. P. Joyner ◽  
Sylvia Berrett ◽  
C. D. H. Boarer ◽  
F. H. Cheong ◽  
...  
Keyword(s):  
Amino Acids ◽  
In Vitro Studies ◽  
Intestinal Malabsorption ◽  
Eimeria Acervulina ◽  
Young Chicks
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