Regulation of Hemoglobin Synthesis in the Blood Islands of Chick Blastodiscs. Tentative Identification of the Stimulatory Transfer RNA as a Minor Alanine-Specific Species

1972 ◽  
Vol 50 (11) ◽  
pp. 1158-1164 ◽  
Author(s):  
S. D. Wainwright ◽  
Lillian K. Wainwright ◽  
H. M. Tsay
Keyword(s):  
Actinomycin D ◽  
Deae Cellulose ◽  
Transfer Rna ◽  
High Concentration ◽  
Adult Chicken ◽  
Tentative Identification ◽  
Hemoglobin Synthesis ◽  
Amino Acid Mixtures ◽  
Specific Species ◽  
A Minor

Transfer RNA, isolated from adult chicken liver and from the membranes associated with the 5- to 6-day embryo, stimulated hemoglobin formation by de-embryonated chick blastodiscs explanted at the 5-somite stage onto medium containing a high concentration of actinomycin D. Preparations acylated specifically with alanine retained this biological activity when oxidized with periodate. The activity of preparations charged with amino acid mixtures lacking alanine was destroyed by periodate treatment.A minor species, representing less than 1% of the total, was largely resolved from the bulk of the alanine-specific tRNAs of chick liver by serial chromatography on benzoylated DEAE-cellulose. The isolated fraction contained the component which stimulated hemoglobin formation by blastodiscs explanted onto medium containing a high concentration of actinomycin.

scholarly journals Structural defects in rat liver deoxyribonucleic acid. Endogenous single-strained regions in comparison with damage induced in vivo by a carcinogen

1979 ◽  
Vol 179 (2) ◽  
pp. 341-352 ◽  
Author(s):  
B W Stewart ◽  
P H Huang ◽  
M J Brian
Keyword(s):  
Rat Liver ◽  
Structural Damage ◽  
Deae Cellulose ◽  
Minor Component ◽  
Structural Defects ◽  
Structural Abnormality ◽  
Denaturation Kinetics ◽  
Limited Digestion ◽  
A Minor

Rat liver DNA may be separated into two fractions by stepwise elution from benzoylated-DEAE-cellulose with NaCl and caffeine solutions respectively. Other studies using bacterical and yeast DNA suggested that the first fraction contains native DNA, whereas the second may exhibit some degree of single-stranded character. In the present experiments, chromatography of DNA was monitored by labelling in vivo with [methyl-3H]thymidine in rats previously subjected to partial hepatectomy. In animals killed up to 1 h after thymidine injection, radioactivity eluted in the second fraction was inversely related to the incorporation time, being greatest when animals were killed 10 min after radioisotope injection. However, for most experiments, animals were allowed to survive 2-4 weeks after surgery before use, analysis being made on non-dividing DNA. Under these conditions, the proportion of caffeine-eluted DNA was decreased by subjecting the preparation to shear, before chromatography. A procedure that resulted in 12% of the recovered radioactivity being eluted with caffeine was adopted for experiments involving comparisons of the two DNA fractions. Under these conditions, cross-contamination could be detected by rechromatography, but this did not preclude distinction being made between the two fractions in terms of DNA structure. NaCl-eluted DNA did not bind to nitrocellulose filters. Caffeine-eluted DNA was retained by the filters and released by washing with 3mM-Tris/HCl, pH9.4. The fractions did not differ in terms of isopycnic centrifugation in CsCl. The NaCl-eluted fraction migrated as a single band in polyacrylamide gels, and this pattern was not modified by prior digestion with Neurospora crassa endonuclease. In contrast, caffeine-eluted DNA contained a minor component having a wide molecular-weight distribution and was subject to limited digestion by the endonuclease. The kinetics of denaturation of NaCi-eluted DNA in the presence of formaldehyde, in common with unfractionated DNA, were consistent with double-stranded structure. The same analysis of caffeine-eluted DNA revealed structural abnormality equivalent to two defects per 10000 base-pairs. The data are consistent with the minor fraction of rat liver DNA, separated by using benzoylated-DEAE-cellulose, containing regions of local denaturation. We previously showed that administration of the hepatocarcinogen dimethylnitrosamine is associated with an increase in the proportion of caffeine-eluted DNA. In terms of most analysis, differences between DNA fraction from nitrosamine-treated rats were similar to differences exhibited by preparations from control animals. However, structural analysis using denaturation kinetics indicated defects in both the NaCl- and caffeine-eluted DNA isolated from nitrosamine-treated rats. The two fractions differed from each other in that caffeine-eluted DNA exhibited a degree of structural damage far greater than that detected in any preparation from control animals...

Reticulocyte transfer RNA and hemoglobin synthesis

Science ◽  
1975 ◽  
Vol 190 (4214) ◽  
pp. 529-535 ◽  
Author(s):  
D. Smith
Keyword(s):  
Transfer Rna ◽  
Hemoglobin Synthesis

scholarly journals Spermine synthesis in the uterus of the ovariectomized rat in response to oestradiol-17β

1972 ◽  
Vol 128 (5) ◽  
pp. 1109-1115 ◽  
Author(s):  
D H. Russell ◽  
J J. Potyraj
Keyword(s):  
Enzyme Activity ◽  
Half Life ◽  
Actinomycin D ◽  
Deae Cellulose ◽  
Ovariectomized Rat ◽  
Enzymic Synthesis ◽  
Hormone Administration

We reported that spermidine and spermine pools in the uterus both doubled within 24h after oestradiol administration to castrated rats (Russell & Taylor, 1971). Now we have studied the enzymic synthesis of spermine (by spermidine-dependent S-adenosyl-l-methionine decarboxylase) and find that the activity of the enzyme(s) involved is elevated soon after hormone administration. Enzyme activity is increased within 4h and is five times that of controls within 24h. Cycloheximide or actinomycin D administered at the time of oestradiol injection completely blocked the increase in enzyme activity. The enzyme involved in spermine synthesis, S-adenosyl-l-methionine decarboxylase, with S-adenosyl-l-methionine and spermidine as required substrates, was partially purified on Sephadex and DEAE-cellulose columns. The decarboxylation of S-adenosyl-l-methionine could not be separated from the transfer of a propylamine moiety from the decarboxylated S-adenosyl-l-methionine to spermidine to form spermine. We were unable also to separate this system from the enzyme that formed spermidine when S-adenosyl-l-methionine and putrescine are used as substrates. Spermidine-stimulated S-adenosyl-l-methionine decarboxylase has an apparent half-life of 60min, identical with the half-life reported for putrescine-stimulated S-adenosyl-l-methionine decarboxylase. These results strongly suggest that the same enzyme(s) operate in the synthesis of both spermidine and spermine.

LEUKOCYTE MOTILITY: II. EFFECT OF ABSENCE OF GLUCOSE IN MEDIUM; EFFECT OF PRESENCE OF DEOXYGLUCOSE, DINITROPHENOL, PUROMYCIN, ACTINOMYCIN D, AND TRYPSIN ON THE RESPONSE TO CHEMOTACTIC SUBSTANCE; EFFECT OF SEGREGATION OF CELLS FROM CHEMOTACTIC SUBSTANCE

1967 ◽  
Vol 45 (2) ◽  
pp. 269-280 ◽  
Author(s):  
Bruce M. Carruthers
Keyword(s):  
Actinomycin D ◽  
Suppressive Effect ◽  
Inhibitory Effect ◽  
Medium Effect ◽  
Inhibitory Effects ◽  
In Vitro Motility ◽  
Directed Motility ◽  
A Minor ◽  
Chemotactic Effect

The random and directed motility of human leukocytes was studied in vitro. Motility was found not to be dependent upon glucose in the medium. 2-Deoxyglucose was found to inhibit all motility completely. Dinitrophenol had a minor suppressive effect on both random and directed motility. Puromycin at 10−3 M and actinomycin D at 10 μg/ml had a disproportionately great inhibitory effect on directed motility, when compared with minor inhibitory effects on random motility. Actinomycin D at 20 μg/ml and trypsin at 0.1 mg/ml were found to inhibit both types of motility almost completely. Segregation of starch from the field of leukocyte motility was found to abolish its chemotactic effect. Restoration of some chemotactic influence was seen if both starch and leukocytes were present in the segregated area.

scholarly journals N-Acetyl-β-d-hexosaminidase component A. Different forms in human tissues and fluids

1973 ◽  
Vol 135 (3) ◽  
pp. 457-462 ◽  
Author(s):  
J. U. Ikonne ◽  
R. B. Ellis
Keyword(s):  
Salt Concentration ◽  
Human Liver ◽  
Deae Cellulose ◽  
Minor Component ◽  
Human Tissues ◽  
Serum Enzyme ◽  
Hexosaminidase A ◽  
Minor Form ◽  
A Minor ◽  
Cellulose Column

1. Hexosaminidase A of human serum was resolved into two components, a minor form with properties identical with those of the single hexosaminidase A component of human liver, and a major form with significantly different properties. 2. The major serum hexosaminidase A form was eluted from a DEAE-cellulose column at a lower salt concentration than that required to elute the liver form. 3. A multiple-pass technique was used to elute the major serum enzyme A from a Sephadex G-150 column before that of liver enzyme A. 4. Clostridium perfringens neuraminidase converted the major component of serum hexosaminidase A into a form that was held less tightly by DEAE-cellulose, but the minor component of the A enzyme of serum, and the A enzyme of liver were not affected. 5. The hexosaminidase A from tears was similar to the A enzyme from serum, whereas those from several human tissues and from urine and lymph were similar to the liver form. 6. The A enzyme from serum may be derived from the A enzyme from liver by glycosylation before secretion.

scholarly journals Specific protein synthesis in isolated epithelium of guinea-pig seminal vesicle. General features

1977 ◽  
Vol 166 (2) ◽  
pp. 155-166 ◽  
Author(s):  
Carlo M. Veneziale
Keyword(s):  
Guinea Pig ◽  
Seminal Vesicle ◽  
Deae Cellulose ◽  
Specific Protein ◽  
Total Soluble Protein ◽  
High Concentration ◽  
Γ Globulins ◽  
Specific Proteins ◽  
Secondary Antibodies ◽  
Sexually Mature

Four intrinsic soluble proteins are synthesized and secreted by sexually mature guinea-pig seminal-vesicle mucosa, which comprises a monolayer of a homogeneous columnar epithelial cell. All four proteins can be extracted readily in 154mm-NaCl from the organ's luminal constituents in which they are present in high concentration. They are referred to as proteins 1, 2, 3 and 4 in order of their elution during DEAE-cellulose column chromatography. Specific primary antibodies were harvested from goats that had been inoculated with the purified vesicular proteins; secondary antibodies were obtained from a donkey inoculated with goat γ-globulins. Double-antibody-immunoprecipitation techniques were developed to precipitate the vesicular proteins. Thus proteins newly synthesized from14C-labelled amino acids could be precipitated and the incorporated radioactivity assessed. Isolated seminal-vesicle mucosa, incubated in only a buffered salt solution containing glucose, readily synthesized the soluble secreted proteins from added [14C]lysine plus [14C]glycine, [14C]histidine plus [14C]glutamate, [14C]glutamine alone and [14C]arginine alone. The rates of incorporation (d.p.m./mg of total soluble protein) of labelled lysine and glycine and of labelled arginine were linear with time over 180min. With the other labelled precursors, rates diminished between 60 and 180min. Labelled protein could be detected after only 10–15min of incubation. Only 4–9% of the newly synthesized protein remained associated with the mucosa; the remainder was found in the cell-free incubation medium. The isolated seminal-vesicle mucosal preparation will provide a unique opportunity to study the synthesis and secretion of abundant cell-specific proteins by this androgen-dependent tissue.

scholarly journals EFFECT OF ACTINOMYCIN D ON RNA SYNTHESIS AND ANTIBODY FORMATION IN THE ANAMNESTIC RESPONSE IN VITRO

1964 ◽  
Vol 119 (6) ◽  
pp. 881-893 ◽  
Author(s):  
J. Donald Smiley ◽  
John G. Heard ◽  
Morris Ziff
Keyword(s):  
Messenger Rna ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Lymphoid Cells ◽  
Limiting Factor ◽  
High Concentration ◽  
Antibody Synthesis ◽  
Anamnestic Response ◽  
Low Concentrations

Antibody synthesis in anamnestic lymphoid cells, measured by incorporation of leucine-C14 into specific antibody, was inhibited at moderate concentrations of actinomycin D. This was accompanied by marked inhibition of synthesis of RNA as measured by incorporation of H3-cytidine monophosphate. However, at low concentrations of actinomycin D, antibody synthesis was unaffected or even increased while RNA synthesis continued to be inhibited. The results obtained suggest that messenger RNA for antibody synthesis, either because it is relatively stable or present in excess, does not become a limiting factor until its synthesis is maximally inhibited. Puromycin, an inhibitor of amino acid coupling, abolished antibody synthesis in low concentration. 6-Mercaptopurine had no effect on the synthesis of antibody or RNA even at high concentration. The data obtained support the view that antibody synthesis follows pathways similar to those utilized for the formation of other types of proteins.

scholarly journals The Effect of Temperature on the Chromatography of Insulin on Deae-Cellulose

1960 ◽  
Vol 13 (1) ◽  
pp. 69 ◽  
Author(s):  
IJ O'donnell ◽  
EOP Thompson
Keyword(s):  
Ionic Strength ◽  
Elution Curve ◽  
Deae Cellulose ◽  
Minor Component ◽  
Ammonia Removal ◽  
Effect Of Temperature ◽  
Bovine Plasma ◽  
Three Samples ◽  
Ph Range ◽  
A Minor

The effect of ionic strength (range 0,15-0, 3), pH (range 7-9), and temperature (range I-25�C) on the chromatographic behaviour of three samples of insulin on diethylaminoethyl (DEAE)-cellulose columns has been studied. These three factors have a similar effect, a decrease of temperature or pH and an increase in ionic strength lowering the elution volume of the protein. The marked effect of temperature is not due to aggregation-disaggregation of the insulin since bovine plasma albumin which does not aggregate reversibly also showed this effect. The desamido component of insulin could not be detected in commercial insulin under the conditions studied but a minor component varying from 2-6 per cent. of the insulin was separated, as well as various amounts of bound ammonia. Removal of zinc from the insulin did not affect the elution curve.

scholarly journals Autotrophic denitrification via nitrate as an effective approach for removal of dissolved sulfide in anaerobic reactors

2020 ◽  
Vol 82 (8) ◽  
pp. 1628-1634
Author(s):  
Jéssyca de Freitas Lima Brito ◽  
Camylla Rachelle Aguiar Araújo ◽  
Gabriel Alfredo Garcia Neto ◽  
André Bezerra dos Santos ◽  
Marcos Erick Rodrigues da Silva ◽  
...  
Keyword(s):  
Organic Matter ◽  
Elemental Sulfur ◽  
Complete Removal ◽  
Minor Effect ◽  
Autotrophic Denitrification ◽  
High Concentration ◽  
Anaerobic Reactors ◽  
Dissolved Sulfide ◽  
Nitrate Supplementation ◽  
A Minor

Abstract This work assessed the effect of adding different concentrations of nitrate (50–300 mg ·L−1) on the removal of dissolved and gaseous sulfide in an anaerobic reactor treating synthetic effluent containing sulfate (100 mg ·L−1) and organic matter (1 g COD·L−1). Autotrophic denitrification, stimulated by the addition of nitrate, was demonstrated to be a very effective approach for removal of dissolved sulfide even in the presence of a high concentration of organic matter (complete removal with 50 mg mg·L−1). However, it had a minor effect on H2S(g). Sulfide remained partially oxidized to elemental sulfur even with excess nitrate (100–300 mg mg·L−1). Therefore, the competition for this electron acceptor between the autotrophic and heterotrophic denitrification pathways may have prevented the conversion of the generated sulfide into sulfate again. No evidence of inhibition of methanogenesis and sulfidogenesis was found during nitrate supplementation.

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