scholarly journals STUDIES ON THE FEULGEN REACTION WITH HISTOCHEMICAL MODEL SYSTEMS

1964 ◽  
Vol 12 (10) ◽  
pp. 758-767 ◽  
Author(s):  
M. J. HARDONK ◽  
P. VAN DUIJN
Keyword(s):  
Crystal Violet ◽  
Deoxyribonucleic Acid ◽  
Spectral Shift ◽  
The Other ◽  
Model Systems ◽  
Cellulose Film ◽  
Feulgen Reaction ◽  
Cell Nuclei ◽  
Amino Groups ◽  
Schiff Reaction

The absorption spectra of Feulgen-stained deoxyribonucleic acid (DNA)-containing membranes have been studied under various conditions. The absorption curve of the Feulgen-stained DNA-cellulose differs from those of the other model systems and is comparable to that of a solution of apurinic acid stained by the Schiff reaction. The spectral shift caused by the reaction of formaldehyde and SO2 with the stained films was used as a criterion for the degree of substitution of the bound pararosaniline. The spectrum of DNA-cellulose, unlike the spectra of the other models, did not show any change under the action of these reagents. This indicates that the three amino groups of pararosaniline are all bound to the DNA in a stained DNA-cellulose film. When the mobility of the apurinic acid chain is restricted, the formation of the trisubstituted product is apparently hindered. Such a situation may be present in DNA-containing polyacrylamide films containing proteins and also in cell nuclei. From a study of the literature it is concluded that the shoulder in the pararosaniline spectrum is most probably caused by a configurational isomer of the dye as indicated by Lewis et al. (16) for crystal violet. To explain the abnormal spectrum of the stained DNA-cellulose, it is proposed that in this case pararosaniline is bound with the three amino groups to three neighbouring deoxyribose groups in the hydrolyzed DNA. Studies with Stuart models of the molecules involved showed that in this case one conformation isomer of pararosaniline can be constructed more easily than the other. This situation may explain the spectrum of the stained DNA-cellulose. The other spectral results could also be interpreted on basis of this hypothesis.

scholarly journals SYNTHESIS AND PROPERTIES OF MODEL SYSTEMS, WITH THEIR USE IN STUDYING THE SCHIFF REACTION IN HISTOCHEMISTRY

1964 ◽  
Vol 12 (7) ◽  
pp. 533-537 ◽  
Author(s):  
M. J. HARDONK ◽  
P. VAN DUIJN
Keyword(s):  
Sulfhydryl Groups ◽  
Phosphate Group ◽  
Model Systems ◽  
Cellulose Derivatives ◽  
Amino Groups ◽  
Cellulose Films ◽  
Reactive Groups ◽  
Schiff Reaction ◽  
Cellulose Membranes ◽  
Covalently Bound

The synthesis of cellulose films containing several reactive groups is described. These films were developed for use as histochemical model systems to study several Schiff-type reactions. The synthesized model systems include cellulose membranes to which amino groups are covalently bound, cellulose membranes containing sulfhydryl groups, and cellulose membranes to which deoxyribonucleotides have been bound by means of their phosphate group. Evidence is given for the structure of these cellulose derivatives.

scholarly journals DNA level in guard cells nuclei of Ornithogalum umbellatum ovary is 2C-4C

2011 ◽  
Vol 77 (3) ◽  
pp. 207-211
Author(s):  
Maria Kwiatkowska ◽  
Katarzyna Rogala ◽  
Katarzyna Popłońska
Keyword(s):  
Dna Content ◽  
Guard Cell ◽  
Flower Development ◽  
Maximum Level ◽  
Guard Cells ◽  
The Other ◽  
Feulgen Reaction ◽  
Cell Nuclei ◽  
Other Hand ◽  
Ovary Growth

The DNA content after Feulgen reaction in the guard cells and epidermis of <em>Omithogalum umbellatum</em> ovary was cytophotometrically measured in different phases of flower development. Only in bud of flowers guard cells DNA content was 2C while in full blown flowers it was higher, between 2C-4C. This observation was supported by autoradiographic studies with 3H-thymidine which was incorporated into guard cell nuclei in the ovary epidermis of newly developed flowers. Thus DNA level in <em>O. umbellatum</em> guard cells was higher than those in other plants described in literature. On the other hand, DNA content in the epidermis cells increased gradually with ovary growth reaching the maximum level of 8C in some cells.

scholarly journals A QUANTITATIVE STUDY OF THE FEULGEN REACTION WITH THE AID OF HISTOCHEMICAL MODEL SYSTEMS

1964 ◽  
Vol 12 (10) ◽  
pp. 752-757 ◽  
Author(s):  
M. J. HARDONK ◽  
P. VAN DUIJN
Keyword(s):  
Absorption Spectra ◽  
Quantitative Study ◽  
Aldehyde Group ◽  
Uv Absorption ◽  
Model Systems ◽  
Feulgen Reaction ◽  
Amino Groups ◽  
Bovine Albumin ◽  
Free Aldehyde ◽  
Quantitative Results

This paper reports some quantitative results with three histochemical model systems. The model systems are DNA-cellulose, deoxyadenylic acid-cellulose and deoxyguanylic acid-cellulose, and DNP-bovine albumin-polyacrylamide. By measuring the UV-absorption of the nucleotide-cellulose after various hydrolysis times, the rates of splitting-off of the purines by HCl were determined. Adenine is found to be split off twice as fast as guanine. The phosphorus/pararosaniline ratio is found to differ considerably from the theoretical value calculated on the assumption that every free aldehyde group reacts with pararosaniline. The differences in the values found for the phosphorus/pararosaniline ratio of the DNA-cellulose and the nucleotide-cellulose can be related to the differences in the absorption spectra of these models, which points to different degrees of substitution at the amino groups of pararosaniline. Addition of protein to DNP in a polyacrylamide film does not alter the phosphorus/pararosaniline ratio.

Peptide synthesis in bone marrow: insulin and thyroxin effects

1962 ◽  
Vol 203 (4) ◽  
pp. 693-696 ◽  
Author(s):  
Thomas F. Necheles
Keyword(s):  
Bone Marrow ◽  
Nucleic Acid ◽  
Peptide Synthesis ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Optimum Concentration ◽  
The Other ◽  
Anabolic Effect ◽  
Oxygen Buffer

Myeloid marrow was rapidly removed from femurs of fasting young rabbits, sectioned, and incubated in Krebs-bicarbonate-CO2-oxygen buffer with appropriate C14-labeled precursors. All manipulations were designed to preserve the architecture of the tissue. After 1 hr the protein or nucleic acid-adenine was isolated and purified. Insulin, 0.01 U/ml added in vitro, stimulated histidine-2(ring)-C14 incorporation into protein by 26 ± 1.4%; alkali-treated insulin was inactive. Thyroxin elicited a 49.4 ± 2.1% stimulation at an optimum concentration of 10–7 m. Triiodothyronine, but not diiodothyronine, also had a significant effect. Insulin increased incorporation of carbon from adenosine-8-C14 into adenine of ribonucleic acid and deoxyribonucleic acid. Thyroxin, on the other hand, was without consistent effect on this process. Thyroxin stimulated significantly the incorporation of C14 of glycine-2-C14 into adenine. The possibility that part of the anabolic effect of thyroxin on bone marrow may arise from a stimulus to incorporation of precursors into purines is suggested.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

Studies Directed Towards the Total Synthesis of Anticapsin and Related Compounds. IV. Competitive Reaction of Amino and Carboxy Substituents in the Halohydrination of 2-Aminobicyclo[2.2.2]oct-5-ene-2-carboxylic Acid Derivatives

1994 ◽  
Vol 47 (12) ◽  
pp. 2221 ◽  
Author(s):  
MJ Crossley ◽  
SR Davies ◽  
TW Hambley
Keyword(s):  
Total Synthesis ◽  
The Other ◽  
X Ray Diffraction ◽  
Amino Groups ◽  
Competitive Reaction ◽  
Trimethylsilyl Group ◽  
X Ray ◽  
Tricyclic Compounds ◽  
Tricyclic Compound ◽  
Related Compounds

Bromohydrination of benzyl (1RS,2SR,4SR)-2-benzyloxycarbonylamino-1-trimethylsilyloxy-bicyclo[2.2.2]oct-5-ene-2-carboxylate (6a) and the (1RS,2RS,4SR)- diastereomer (6b) with N- bromoacetamide in aqueous dioxan has been investigated. These reactions are highly regio-and stereo-selective and give the corresponding bromohydrins (9a) and (9b), but in moderate to low yield. These bromohydrins have the necessary stereochemistry for conversion into anticapsin. The other products from the reaction are tricyclic compounds formed by capture of the anti- bromonium cation intermediates or resultant bromohydrins by interaction with the proximal protected carboxy and amino groups within the molecules. Thus the carbolactone (11) is formed from the endo -adduct (6a), and the carbonimidic acid derivative (12) and the cyclic urethane (13) are formed from the exo-adduct (6b). Cleavage of the trimethylsilyl group from the tricyclic compound (12) gives benzyl (1RS,2RS,3RS,7RS,8RS)-5-benzyloxy-2-bromo-8-hydroxy-4-oxa-6-azatricyclo[5.3.1.03,8]undec-5-ene-7-carboxylate(14), the structure of which was determined by X-ray diffraction methods and refined to a residual of 0.035 for 1549 independent observed reflections. The crystals of (14) are monoclinic, P21/c, a 12.954(3), b 6.197(3), c 26.784(7) Ǻ, β 95.33(2)°, Z 4. Reactions attempting to generate iodohydrins from the alkenes (6) were also highly regioselective and gave detrimethylsilylated iodo analogues of (11) and (13).

Effects of caffeine treatment on aged porcine oocytes: parthenogenetic activation ability, chromosome condensation and development to the blastocyst stage after somatic cell nuclear transfer

Zygote ◽  
2005 ◽  
Vol 13 (4) ◽  
pp. 335-345 ◽  
Author(s):  
Masaki Iwamoto ◽  
Akira Onishi ◽  
Dai-ichiro Fuchimoto ◽  
Tamas Somfai ◽  
Shun-ichi Suzuki ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Blastocyst Stage ◽  
The Other ◽  
Control Group ◽  
Blastocyst Formation ◽  
Cell Nuclei ◽  
Somatic Nucleus ◽  
Membrane Breakdown ◽  
Caffeine Treatment ◽  
M Phase

The possibility of using aged porcine oocytes treated with caffeine, which inhibits the decrease in M-phase promoting factor activity, for pig cloning was evaluated. Cumulus–oocyte complexes (COCs) were cultured initially for 36h and subsequently with or without 5mM caffeine for 24h (in total for 60h: 60CA+ or 60CA– group, respectively). As a control group, COCs were cultured for 48h without caffeine (48CA–). The pronuclear formation rates at 10h after electrical stimulation in the 60CA+ and 60CA– groups decreased significantly (p<0.05) compared with the 48CA– group. However, the fragmentation rate was significantly higher (p<0.05) in the 60CA– group than in the 60CA+ and 48CA– groups. When the stimulated oocytes were cultured for 6 days, the 60CA+ group showed significantly lower blastocyst formation and higher fragmentation or degeneration rates (p<0.05) than the 48CA– group. However, the number of total cells in blastocysts was not affected by maturation period or caffeine treatment. When somatic cell nuclei were injected into the non-enucleated oocytes and exposed to cytoplasm for a certain duration (1–11h) before the completion of maturation (48 or 60h), the rate of nuclear membrane breakdown after exposure to cytoplasm for 1–2h in the 60CA– oocytes was significantly lower (p;<0.05) than in the other experimental groups. The rate of scattered chromosome formation in the same 60CA– group tended to be lower (p=0.08) than in the other groups. After the enucleation and transfer of nuclei, blastocyst formation rates in the 60CA+ and 60CA– groups were significantly lower (p<0.05) than in the 48CA– group. Blastocyst quality did not differ among all the groups. These results suggest that chromosome decondensation of the transplanted somatic nucleus is affected by both the duration of exposure to cytoplasm and the age of the recipient porcine oocytes, and that caffeine treatment promotes nuclear remodelling but does not prevent the decrease in the developmental ability of cloned embryos caused by oocyte aging.

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