scholarly journals ENZYMATIC AND CHROMOSOMAL CHARACTERIZATION OF HELA VARIANTS

1969 ◽  
Vol 41 (3) ◽  
pp. 806-815 ◽  
Author(s):  
R. H. Bottomley ◽  
A. L. Trainer ◽  
M. J. Griffin
Keyword(s):  
Alkaline Phosphatase ◽  
Chromosome Number ◽  
Cell Line ◽  
Lactic Dehydrogenase ◽  
Single Chromosome ◽  
Modal Chromosome Number ◽  
Cell Strains ◽  
Hela S3 ◽  
Glucose 6 Phosphate Dehydrogenase

Seven strains of HeLa cells have been characterized by the number of chromosomes and the activity of the enzymes alkaline phosphatase, glucose-6-phosphate dehydrogenase, 6-phosphogluconic dehydrogenase, and lactic dehydrogenase. All seven strains were found to differ as to chromosome numbers and enzyme levels despite the fact that two strains were called HeLa and three were called HeLa S3. Three strains were found to have a stemline in which greater than 60% of the cells demonstrated a single chromosome number, and this characteristic was stable for at least 6 months. A nomenclature for these clones has been suggested by the use of the stemline chromosome number as a subscript following HeLa. These three clones were, therefore, designated HeLa65, HeLa71, and HeLa75. Karyotypes were made of the stemlines of these clones and were compared with enzyme levels. Alkaline phosphatase showed the greatest variation from cell line to cell line with a 200-fold difference in levels, whereas glucose-6-phosphate dehydrogenase showed variation in activity over a 12-fold range, lactic dehydrogenase over an 8-fold range, and 6-phosphogluconic dehydrogenase over a 2-fold range. It is suggested that human cell strains can be used for biochemical studies if they are cloned and if the clones are relatively stable at least with respect to modal chromosome number and karyotype.

THE ACTIVITY OF ENZYMES IN THE RABBIT UTERUS AND EFFECT OF PROGESTERONE AND OESTRADIOL

1969 ◽  
Vol 43 (2) ◽  
pp. 167-174 ◽  
Author(s):  
R. N. MURDOCH ◽  
I. G. WHITE
Keyword(s):  
Alkaline Phosphatase ◽  
Enzyme Activity ◽  
Succinic Dehydrogenase ◽  
Lactic Dehydrogenase ◽  
Glutamate Oxaloacetate Transaminase ◽  
Oestrogen Treatment ◽  
Uterine Endometrium ◽  
Uterine Fluid ◽  
Activity Of Enzymes ◽  
Glucose 6 Phosphate Dehydrogenase

SUMMARY The activity of several enzymes has been measured in the uterine endometrium of the rabbit during oestrus and pseudopregnancy and after injecting oestradiol benzoate or progesterone 28 days after ovariectomy. The enzyme activity of the uterine fluid has been determined during oestrus and the effect of uterine ligation studied. Progesterone and the induction of pseudopregnancy stimulated succinic dehydrogenase (SDH) and glucose-6-phosphate dehydrogenase (GDH) activity and depressed amylase and lactic dehydrogenase (LDH) activity. In ovariectomized does, glutamate-oxaloacetate transaminase (GOT) activity increased after the injection of progesterone. Progesterone also stimulated endometrial phosphatase after ovariectomy but, when given after a period of oestrogen treatment, it limited the even greater response of acid and alkaline phosphatase to oestrogen; the activity then attaining the same level as when progesterone alone was given. SDH, GDH and glycerylphosphorylcholine (GPC) diesterase could not be detected in uterine fluid but amylase and alkaline phosphatase were in greater concentration than in the endometrium. GPC diesterase was, however, found to be present in uterine tissue. Ligation of the uterus did not significantly alter the enzyme activity of the endometrium.

scholarly journals Characterization of human foetal intestinal alkaline phosphatase. Comparison with the isoenzymes from the adult intestine and human tumour cell lines

1983 ◽  
Vol 211 (3) ◽  
pp. 553-558 ◽  
Author(s):  
C M Behrens ◽  
C A Enns ◽  
H H Sussman
Keyword(s):  
Alkaline Phosphatase ◽  
Cell Line ◽  
Tumour Cell ◽  
Tumour Cell Line ◽  
Subunit Structure ◽  
Human Tumour ◽  
Intestinal Alkaline Phosphatase ◽  
Alkaline Phosphatases ◽  
Inhibition Studies

The molecular structure of human foetal intestinal alkaline phosphatase was defined by high-resolution two-dimensional polyacrylamide-gel electrophoresis and amino acid inhibition studies. Comparison was made with the adult form of intestinal alkaline phosphatase, as well as with alkaline phosphatases isolated from cultured foetal amnion cells (FL) and a human tumour cell line (KB). Two non-identical subunits were isolated from the foetal intestinal isoenzyme, one having same molecular weight and isoelectric point as placental alkaline phosphatase, and the other corresponding to a glycosylated subunit of the adult intestinal enzyme. The FL-cell and KB-cell alkaline phosphatases were also found to contain two subunits similar to those of the foetal intestinal isoenzyme. Characterization of neuraminidase digests of the non-placental subunit showed it to be indistinguishable from the subunits of the adult intestinal isoenzyme. This implies that no new phosphatase structural gene is involved in the transition from the expression of foetal to adult intestinal alkaline phosphatase, but that the molecular changes involve suppression of the placental subunit and loss of neuraminic acid from the non-placental subunit. Enzyme-inhibition studies demonstrated an intermediate response to the inhibitors tested for the foetal intestinal, FL-cell and KB-cell isoenzymes when compared with the placental, adult intestinal and liver forms. This result is consistent with the mixed-subunit structure observed for the former set of isoenzymes. In summary, this study has defined the molecular subunit structure of the foetal intestinal form of alkaline phosphatase and has demonstrated its expression in a human tumour cell line.

scholarly journals Karyotypic characterization of Prochilodus mariae, Semaprochilodus kneri and S. laticeps(Teleostei: Prochilodontidae) from Caicara del Orinoco, Venezuela

2003 ◽  
Vol 1 (1) ◽  
pp. 47-52 ◽  
Author(s):  
Claudio Oliveira ◽  
Mauro Nirchio ◽  
Ángel Granado ◽  
Sara Levy
Keyword(s):  
Chromosome Number ◽  
Chromosome Pair ◽  
Nucleolus Organizer ◽  
Single Chromosome ◽  
Nucleolus Organizer Regions ◽  
The Family ◽  
Structural Chromosome ◽  
Almost All ◽  
Freshwater Environments

Fish of the family Prochilodontidae are considered one of the most important components of commercial and subsistence fishery in freshwater environments in South America. This family consists of 21 species and three genera. In the present study, the karyotypes of Prochilodus mariae, Semaprochilodus kneri, and S. laticeps from Caicara del Orinoco, Bolivar State, Venezuela were studied. The species P. mariae, S. kneri and S. laticeps exhibited 2n=54 chromosomes (40 metacentric and 14 submetacentric), a single chromosome pair with nucleolus organizer regions, and a large amount of heterochromatin found at centromeric and pericentromeric positions in almost all chromosomes. The P. mariae specimens studied displayed 0 to 3 supernumerary microchromosomes. The data obtained here confirm the conservative nature of the chromosome number and morphology of Prochilodontidae and reinforce the hypothesis that small structural chromosome rearrangements were the main cause of the karyotypic diversification seen in this group.

scholarly journals Characterization of a new megakaryocytic cell line: the Dami cell

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 1968-1977 ◽  
Author(s):  
SM Greenberg ◽  
DS Rosenthal ◽  
TA Greeley ◽  
R Tantravahi ◽  
RI Handin
Keyword(s):  
Cell Line ◽  
Chromosomal Abnormalities ◽  
Light And Electron Microscopy ◽  
Ploidy Levels ◽  
Megakaryoblastic Leukemia ◽  
Modal Chromosome Number ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Megakaryocytic Cell

Abstract A new human megakaryocytic cell line (Dami) has been established from the blood of a patient with megakaryoblastic leukemia. The Dami cells grow primarily in suspension with a doubling time of 24 to 30 hours. By light and electron microscopy, the Dami cells range in size from 12 to 120 micron in diameter and have lobulated nuclei characteristic of megakaryocytes. At least 89% of the cells react with monoclonal antibodies against platelet glycoproteins (GP) Ib and IIB/IIIa, and glycophorin. The cells do not react with antibodies against lymphoid, monocyte, granulocyte, or macrophage antigens. Thirteen percent of the cells become polyploid, spontaneously achieving greater than 4N DNA ploidy levels. In response to phorbol myristate acetate (PMA), the proportion of cells with ploidy levels greater than 4N increased threefold and could be separated into discrete ploidy groups. PMA also increased the expression of GPIb, the GPIIb/GPIIIa complex,l and von Willebrand factor. Cytogenetic analysis revealed a human male hyperdiploid karyotype with a modal chromosome number of 54 to 64 and several consistent clonal chromosomal abnormalities. These included a partial deletion of chromosome 5 and a translocation involving chromosome 3. In contrast to other megakaryocytic cell lines in which only a small portion of the cells express the megakaryocytic phenotype, nearly all of the Dami cells express platelet glycoproteins. Thus, the Dami cells provide a superior model in which to study human megakaryocyte biochemistry and differentiation.

scholarly journals Characterization of a new megakaryocytic cell line: the Dami cell

Blood ◽  
1988 ◽  
Vol 72 (6) ◽  
pp. 1968-1977 ◽  
Author(s):  
SM Greenberg ◽  
DS Rosenthal ◽  
TA Greeley ◽  
R Tantravahi ◽  
RI Handin
Keyword(s):  
Cell Line ◽  
Chromosomal Abnormalities ◽  
Light And Electron Microscopy ◽  
Ploidy Levels ◽  
Megakaryoblastic Leukemia ◽  
Modal Chromosome Number ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Megakaryocytic Cell

A new human megakaryocytic cell line (Dami) has been established from the blood of a patient with megakaryoblastic leukemia. The Dami cells grow primarily in suspension with a doubling time of 24 to 30 hours. By light and electron microscopy, the Dami cells range in size from 12 to 120 micron in diameter and have lobulated nuclei characteristic of megakaryocytes. At least 89% of the cells react with monoclonal antibodies against platelet glycoproteins (GP) Ib and IIB/IIIa, and glycophorin. The cells do not react with antibodies against lymphoid, monocyte, granulocyte, or macrophage antigens. Thirteen percent of the cells become polyploid, spontaneously achieving greater than 4N DNA ploidy levels. In response to phorbol myristate acetate (PMA), the proportion of cells with ploidy levels greater than 4N increased threefold and could be separated into discrete ploidy groups. PMA also increased the expression of GPIb, the GPIIb/GPIIIa complex,l and von Willebrand factor. Cytogenetic analysis revealed a human male hyperdiploid karyotype with a modal chromosome number of 54 to 64 and several consistent clonal chromosomal abnormalities. These included a partial deletion of chromosome 5 and a translocation involving chromosome 3. In contrast to other megakaryocytic cell lines in which only a small portion of the cells express the megakaryocytic phenotype, nearly all of the Dami cells express platelet glycoproteins. Thus, the Dami cells provide a superior model in which to study human megakaryocyte biochemistry and differentiation.

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