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1.
Cell Biol Int ; 31(10): 1214-9, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17517524

ABSTRACT

A non-transformed (Vero) and two tumor cell lines (HepG2 and MCF-7) were treated with 10nM to 100 microM formaldehyde. Lower doses (10nM to 10 microM) enhanced the viability of the cultured cells, measured by MTT assay. Higher doses (75-100 microM) decreased viability of the cells by 50% or more. The 100 microM concentration of HCHO has been chosen for combination treatment of the three cell lines with a series of concentrations (0.2-100 microM) of resveratrol, a phytoestrogen occurring in various fruits. Resveratrol decreased the cytotoxicity of formaldehyde depending on cell line and point of time, especially in case of MCF-7 cells at 24 and 72 h, Vero cells at 24h and HepG2 cells at 48 h after treatment. Possible modes of interactions are discussed, considering the role of resveratrol in formaldehyde metabolism and also the estrogen receptor positivity of MCF-7 cells.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Cell Survival/drug effects , Disinfectants/pharmacology , Formaldehyde/pharmacology , Stilbenes/pharmacology , Angiogenesis Inhibitors/pharmacology , Animals , Breast Neoplasms/drug therapy , Breast Neoplasms/pathology , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Cells, Cultured/drug effects , Chlorocebus aethiops , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Receptors, Estrogen/metabolism , Resveratrol , Vero Cells/drug effects
3.
J Toxicol Environ Health A ; 62(5): 333-47, 2001 Mar 09.
Article in English | MEDLINE | ID: mdl-11261896

ABSTRACT

Many molecular investigations of colorectal cancer (CRC) have suggested that the accumulation of specific mutations in proto-oncogenes and tumor suppressor genes regulating cell growth via signal transduction trigger the stagewise progression to malignancy. In this study, the frequency, location, and type of mutations of the K-ras proto-oncogene exon I and p53 tumor suppressor gene exons 5-8 were analyzed in colorectal carcinomas of 65 patients from Central Europe, using polymerase chain reaction (PCR)-cold single-strand conformation polymorphism (SSCP) screening and direct sequencing. The incidence of K-ras activating mutations in these Central European samples was lower (25%) compared to that obtained in American and western European populations (40-50% at least), while the incidence of p53 inactivating mutations was similar (58%). These results suggest that some other genetically linked mechanisms may play a role in CRC development and progression, and hence K-ras and p53 mutations cannot be considered to be universal genetic markers for CRC.


Subject(s)
Colorectal Neoplasms/genetics , Genes, ras/genetics , Tumor Suppressor Protein p53/genetics , Colorectal Neoplasms/metabolism , DNA Primers , DNA, Neoplasm/genetics , DNA, Neoplasm/isolation & purification , Europe , Exons/genetics , Humans , Introns/genetics , Polymorphism, Single-Stranded Conformational , Proto-Oncogene Mas , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA
4.
J Physiol Paris ; 94(1): 57-62, 2000.
Article in English | MEDLINE | ID: mdl-10761690

ABSTRACT

Numerous studies have reported diverse effects of gut-derived regulatory peptides on growth of the normal pancreas, pancreatic neoplasms induced experimentally in animals, and pancreatic cancer cell lines, but the results of these investigations are rather controversial. The stimulatory effect of epidermal growth factor (EGF) on cell proliferation of pancreatic cell lines is well established. Whether this action can be modulated by somatostatin is not clear. Furthermore, it is not certain whether another regulatory peptide, cholecystokinin (CCK), affects the proliferation of these cells. In the present study we investigated the presence of CCK-A and CCK-B, as well as somatostatin-2 (SSTR2) receptors by RT-PCR, and studied the actions of EGF, CCK and octreotide on DNA synthesis in the human pancreatic adenocarcinoma cell line Capan-2. Octreotide, a long-acting somatostatin analogue was used as somatostatin agonist. Cells were cultured in RPMI-1640 medium. They were incubated in serum free medium containing 0.2% BSA in the absence (control) or the presence of the peptides. [3H]-thymidine incorporation into DNA was measured after 48 h of incubation. By means of RT-PCR analysis we were able to demonstrate SSTR2 expression, but not CCK-A or CCK-B receptor mRNA in Capan-2 cells. DNA synthesis evaluated by [3H]-thymidine incorporation was found to be increased by 45.2 +/- 5.6% in response to EGF (10(-8) M) and decreased by 11.7 +/- 2.6% to octreotide (10(-8) M) compared to controls (P < 0.01). The increase in [3H]-thymidine incorporation was significantly lower when EGF treatment was combined with octreotide administration (10.1 +/- 2.5% over control). In the concentration range of 10(-11)-10(-8) M, CCK did not alter significantly the incorporation of [3H]-thymidine into DNA in Capan-2 cells. In conclusion, these data support a role for EGF as a growth factor for the human pancreatic cancer cell Capan-2. Somatostatin may play an important role in regulating cell proliferation in Capan-2 cells both under basal, and growth factor-stimulated conditions. Our results suggest, however, that CCK receptors are not expressed, and CCK does not affect cell proliferation in this transformed pancreatic cell line.


Subject(s)
Adenocarcinoma/pathology , Epidermal Growth Factor/pharmacology , Octreotide/pharmacology , Pancreatic Neoplasms/pathology , Adenocarcinoma/metabolism , Cell Division/drug effects , Cholecystokinin/pharmacology , DNA/biosynthesis , Humans , Pancreatic Neoplasms/metabolism , Protein Isoforms/metabolism , Receptor, Cholecystokinin A , Receptor, Cholecystokinin B , Receptors, Cholecystokinin/metabolism , Receptors, Somatostatin/metabolism , Thymidine/metabolism , Tumor Cells, Cultured/pathology
5.
Leukemia ; 14(4): 706-11, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10764158

ABSTRACT

We have studied the frequency of p53 mutations in genomic DNA extracted from peripheral blood or the spleen of 61 patients with hairy cell leukemia using PCR-SSCP and automated cycle sequencing. We identified exon 5-8 mutations in 17 cases, corresponding to a frequency of 28%. In four cases, mutations were localized in exon 5; one patient with atypical HCL had a mutation in exon 6 at the 3' boundary; five cases showed mutations in exon 7, while exon 8 was found to be mutated in seven cases. The mutations found could be divided into three major categories: structural (n=9), inactivating (n= 6), and neutral (n= 2) mutations. None of the three transitions found occurred at CpG dinucleotides. The rate of p53 mutations found in this large cohort of HCL patients is unexpectedly high as in other non-Hodgkin lymphomas p53 mutations predict for poor treatment outcome. The character of the mutations we have found is entirely different from that described in other hematologic malignancies.


Subject(s)
Genes, p53 , Leukemia, Hairy Cell/genetics , Mutation , Adult , Aged , Amino Acid Substitution , Cell Cycle , Codon , Cohort Studies , CpG Islands , DNA Mutational Analysis , DNA, Neoplasm/blood , DNA, Neoplasm/genetics , Female , Humans , Leukemia, Hairy Cell/mortality , Leukemia, Hairy Cell/pathology , Lymphoma, Non-Hodgkin/genetics , Lymphoma, Non-Hodgkin/mortality , Male , Middle Aged , Point Mutation , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Prognosis , Sequence Deletion , Spleen/chemistry
6.
Biol Chem ; 381(2): 95-105, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10746740

ABSTRACT

Latent episomal genomes of Epstein-Barr virus, a human gammaherpesvirus, represent a suitable model system for studying replication and methylation of chromosomal DNA in mammals. We analyzed the methylation patterns of CpG dinucleotides in the latent origin of DNA replication of Epstein-Barr virus using automated fluorescent genomic sequencing of bisulfite-modified DNA samples. We observed that the minimal origin of DNA replication was unmethylated in 8 well-characterized human cell lines or clones carrying latent Epstein-Barr virus genomes as well as in a prototype virus producer marmoset cell line. This observation suggests that unmethylated DNA domains can function as initiation sites or zones of DNA replication in human cells. Furthermore, 5' from this unmethylated region we observed focal points of de novo DNA methylation in nonrandom positions in the majority of Burkitt's lymphoma cell lines and clones studied while the corresponding CpG dinucleotides in viral genomes carried by lymphoblastoid cell lines and marmoset cells were completely unmethylated. Clustering of highly methylated CpG dinucleotides suggests that de novo methylation of unmethylated double-stranded episomal viral genomes starts at discrete founder sites in vivo. This is the first comparative high-resolution methylation analysis of a latent viral origin of DNA replication in human cells.


Subject(s)
DNA Methylation , DNA Replication , Genome, Viral , Herpesvirus 4, Human/genetics , Replication Origin/genetics , Virus Replication , Base Sequence , Binding Sites/genetics , Burkitt Lymphoma/genetics , Cell Line , Cytosine/metabolism , DNA Transposable Elements , Electronic Data Processing , Epstein-Barr Virus Nuclear Antigens/genetics , Humans , Lymphocytes , Molecular Sequence Data , Repetitive Sequences, Nucleic Acid , Sequence Analysis, DNA
7.
Biol Chem ; 379(4-5): 545-7, 1998.
Article in English | MEDLINE | ID: mdl-9628350

ABSTRACT

Mutations of the p53 gene may alter the specific regulatory domains of the protein. We examined the conserved domains III, IV and V by SSCP using PCR primers covering exons 5, 6, 7 and 8 from hairy cell leukemia (HCL), polyps, colorectal and gastric carcinomas. A low rate of p53 mutations was detected in HCL and polyps. These mutations may predict the risk of malignant development. However, multiple mutations were a frequent occurrence in tumors. Sequence analysis of our samples did not demonstrate the high frequency of transition mutations (C-->T) that would be predicted if the major course of p53 mutations is deamination of 5-methylcytosine (5mC). Rather, most mutations were found to be single base insertions or deletions.


Subject(s)
Colonic Polyps/genetics , DNA Methylation , Genes, p53 , Leukemia, Hairy Cell/genetics , Mutation , Stomach Neoplasms/genetics , Colonic Polyps/pathology , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Deamination , Humans , Leukemia, Hairy Cell/pathology , Mutagenesis, Insertional , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Sequence Deletion , Stomach Neoplasms/pathology
8.
FEBS Lett ; 419(2-3): 181-5, 1997 Dec 15.
Article in English | MEDLINE | ID: mdl-9428630

ABSTRACT

By applying the 'recognition mask' strategy to 300 mammalian sequences containing NotI sites we demonstrated that 5' ends of genes are highly enriched in NotI sites. A NotI linking clone NL2-252 (D3S1678) containing transferrin receptor (TFRC) gene was used as an initial point for chromosomal jumping. One of the jumping clones, J21-045 traverses 210 kbp and links NL2-252 to NL26 (D3S1632), a NotI linking clone containing highly polymorphic sequences. The TFRC gene was mapped to 3q29, close to the telomeric marker D3S2344, by linkage analysis, a panel of hybrid cell lines, GeneBridge 4 panel and FISH. Clone NLM-007 (D3S4302) was found to contain ras-homologous gene RAB7. By FISH and a panel of hybrid cell lines this gene was mapped to 3q21. This region is of particular interest due to frequent rearrangements in different types of leukemia. Clone L2-081 (D3S4283) containing new member of ubiquitin-specific proteases (HAUSP gene) was localized in 3p21 inspiring further investigation of involvement of this gene in development of lung and renal carcinomas.


Subject(s)
Chromosome Mapping , Chromosomes, Human, Pair 3 , Endopeptidases/genetics , GTP-Binding Proteins/genetics , Gene Library , Genome, Human , Leukemia/genetics , Neoplasms/genetics , Receptors, Transferrin/genetics , rab GTP-Binding Proteins , Cloning, Molecular , Gene Rearrangement , Humans , Molecular Sequence Data , Ubiquitin Thiolesterase , Ubiquitin-Specific Peptidase 7 , rab7 GTP-Binding Proteins
9.
Acta Physiol Hung ; 83(4): 323-32, 1995.
Article in English | MEDLINE | ID: mdl-8863895

ABSTRACT

The maximal capacity of low affinity ouabain binding sites in kidney medulla was found to be increased by 20 +/- 3.8% after 2 weeks, and by 35 +/- 4.5% in 4 weeks diabetes. However, in kidney cortex no similar changes could be detected. Western blot analysis of Na+/K(+)-ATPase subunits in kidney medulla indicated a significant enhancement of both the alpha 1 and beta 1 subunit in two and four weeks diabetic rats (alpha 1: 35 +/- 3.1, 51 +/- 5.8% and beta 1: 31.3 +/- 5.2 and 43.2 +/- 6.8%, respectively). However, kidney cortex showed no significant change in any condition tested. In diabetes we could detect a significant change only in the medulla in case of the b subunit mRNA transcript, which showed 1.69 +/- 0.59 and 2.89 +/- 0.81 times increased in two and four weeks diabetic state, respectively. There was no change in the alpha 1 subunit mRNA abundance. Insulin treatment of diabetic animals did not result in a complete reversal of diabetes-induced changes in ouabain binding capacity or in the amount of Na+/K(+)-ATPase alpha 1 and beta 1 subunit protein and mRNA levels. Our data indicate a good correlation between changes in low affinity ouabain binding capacity and the level of alpha 1 isoform in diabetic rats, and suggest an important role of the b subunit in the regulation of enzyme quantity.


Subject(s)
Diabetes Mellitus, Experimental/metabolism , Kidney/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Streptozocin/pharmacology , Animals , Body Weight/drug effects , Disease Models, Animal , Male , Rats
10.
Acta Physiol Hung ; 83(4): 333-42, 1995.
Article in English | MEDLINE | ID: mdl-8863896

ABSTRACT

The 78 kDa glucose-regulated protein (grp78) is an abundant member of the 70 kDa molecular chaperone family in the lumen of the endoplasmic reticulum participating in the quality control of secretory proteins. In the present paper we have analysed the synthesis and level of grp78 in livers of control, streptozotocin-diabetic, and the spontaneously diabetic Zucker rats. The level of grp78 mRNA significantly decreased in streptozotocin-diabetic rats. The effect was reversed by insulin treatment. In case of Zucker rats we did not detect any significant change in grp78 mRNA, grp78 protein level showed opposite changes being essentially unchanged in streptozotocin-diabetes and significantly reduced in Zucker rats. Autoradiograms of Ca-dependent phosphorylation of postmitochondrial supernatants of control and streptozotocin-diabetic livers indicated no significant changes in the 70 kDa region. Decrease in the availability of grp78 may participate in the attenuation of hepatic protein secretion in diabetes.


Subject(s)
Carrier Proteins/metabolism , Diabetes Mellitus, Experimental/metabolism , Glucose/metabolism , Heat-Shock Proteins , Liver/metabolism , Molecular Chaperones/metabolism , Animals , Disease Models, Animal , Endoplasmic Reticulum Chaperone BiP , Male , Rats , Rats, Sprague-Dawley
11.
Biochem Biophys Res Commun ; 188(3): 1191-7, 1992 Nov 16.
Article in English | MEDLINE | ID: mdl-1332712

ABSTRACT

The Na+/K(+)-ATPase activity and expression of mRNAs encoding alpha and beta subunits of the enzyme were examined in rat myometrium at different stages of pregnancy. The enzyme activity appeared to increase during the pregnancy and reached the highest value at the 17th day. Northern blot analysis of total RNA isolated from the same myometrial samples shows the expressions of alpha 1, alpha 3 and beta mRNAs. A2 mRNA was not detectable. By the progression of pregnancy until the 17th day the expression of all the three kinds of mRNA increased. The change in the abundance of mRNA-beta was much more higher (12-fold) than that of mRNA-alpha 1 and -alpha 3 (4 and 2.5-fold, respectively). Furthermore, the expression of mRNA-beta sharply decreased after the 17th day, while the level of alpha subunits mRNAs barely changed. We conclude that transcriptional regulation of the Na+/K(+)-ATPase subunits could be different during this physiological process.


Subject(s)
Myometrium/metabolism , Pregnancy, Animal/metabolism , RNA, Messenger/analysis , Sodium-Potassium-Exchanging ATPase/biosynthesis , Transcription, Genetic , Animals , Female , Pregnancy , Rats , Rats, Inbred Strains , Rats, Wistar
12.
J Gen Virol ; 73 ( Pt 7): 1687-92, 1992 Jul.
Article in English | MEDLINE | ID: mdl-1321209

ABSTRACT

The genome of Epstein-Barr virus (EBV) codes for two non-translated small RNA molecules, EBER 1 and 2. We found that both EBERs are expressed in the major EBV-carrying cell types, group I and III Burkitt's lymphoma (BL) cell lines, lymphoblastoid cell lines (LCLs) and in two nude mouse-passaged nasopharyngeal carcinoma (NPC) tumours. The relative amount of EBER 1 and EBER 2 varied in different host cells but did not correlate with the cellular phenotype. The EBER coding and flanking sequences were predominantly hypomethylated at HpaII sites not only in LCLs which usually carry hypomethylated EBV genomes but also in BL and NPC cell lines harbouring EBV episomes that are highly methylated in other regions. Thus, the EBER transcription units, actively transcribed by RNA polymerase III in the major EBV-carrying cell types, represent a methylation-free region in the EBV genome similarly to regulatory sequences of the latent membrane protein gene when the latter is transcribed by RNA polymerase II.


Subject(s)
Gene Expression , Herpesvirus 4, Human/genetics , Proteins/genetics , RNA Polymerase III/physiology , RNA-Binding Proteins , Transcription, Genetic , Animals , Burkitt Lymphoma/microbiology , Cell Line , Methylation , Mice , Mice, Nude
13.
Genomics ; 11(4): 1030-9, 1991 Dec.
Article in English | MEDLINE | ID: mdl-1783374

ABSTRACT

A novel procedure for construction of jumping libraries is described. The essential features of this procedure are as follows: (1) two diphasmid vectors (lambda SK17 and lambda SK22) are simultaneously used in the library construction to improve representativity, (2) a partial filling-in reaction is used to eliminate cloning of artifactual jumping clones and to obviate the need for a selectable marker. The procedure has been used to construct a representative human NotI jumping library (220,000 independent recombinant clones) from the lymphoblastoid cell line CBMI-Ral-STO, which features a low level of methylation of its resident EBV genomes. A human chromosome 3-specific NotI jumping library (500,000 independent recombinant clones) from the human chromosome 3 x mouse hybrid cell line MCH 903.1 has also been constructed. Of these recombinant clones 50-80% represent jumps to the neighboring cleavable NotI site. With our previously published method for construction of linking libraries this procedure makes a new genome mapping strategy feasible. This strategy includes the determination of tagging sequences adjacent to NotI sites in random linking and jumping clones. Special features of the lambda SK17 and lambda SK22 vectors facilitate such sequencing. The STS (sequence tagged site) information obtained can be assembled by computer into a map representing the linear order of the NotI sites for a chromosome or for the entire genome. The computerized mapping data can be used to retrieve clones near a region of interest. The corresponding clones can be obtained from the panel of original clones, or necessary probes can be made from genomic DNA by PCR.


Subject(s)
Chromosome Mapping/methods , Chromosomes, Human, Pair 3 , Gene Library , Genome, Human , Animals , Cloning, Molecular , Deoxyribonucleases, Type II Site-Specific/metabolism , Electrophoresis, Gel, Pulsed-Field , Genes, Tumor Suppressor , Genetic Vectors , Humans , Hybrid Cells , Mice , Tumor Cells, Cultured
14.
Genes Chromosomes Cancer ; 3(5): 403-6, 1991 Sep.
Article in English | MEDLINE | ID: mdl-1797089

ABSTRACT

Deletions of the short arm of chromosome 3 and associated allele losses have been reported in the majority of sporadic renal cell carcinomas (RCC). On the basis of the combined cytogenetic and molecular data, it is reasonable to assume that a putative RCC locus, which contributes to tumor development by its loss, is located telomerically of the D3F15S2 site. Using H3E4, a D3F15S2-specific probe, we have isolated a cDNA clone (cl.4-2), and a sequence comparison revealed that the cDNA clone corresponds to the human acyl-peptide hydrolase gene. The gene is fairly universally expressed, but in RCC biopsies its expression is severely reduced, compared to the normal kidney. Cl.4-2 was used for in situ hybridization on metaphase chromosomes prepared from an Epstein-Barr virus (EBV) transformed lymphoblastoid cell line, derived from a t(3;8) (p14.2;q24.1) carrying member of the RCC family described by Cohen et al. in 1979 (N Engl J Med: 301:592-595). Carriers of this translocation regularly develop RCC by middle age. We now report that D3F15S2 is localized on the telomeric side of the constitutional breakpoint, in 3p21. The region of 3p affected by this familial translocation is thus not identical with the region of 3p most frequently deleted in sporadic RCC.


Subject(s)
Carcinoma, Renal Cell/genetics , Chromosome Deletion , Chromosomes, Human, Pair 3 , Kidney Neoplasms/genetics , Alleles , Chromosome Banding , Humans , Karyotyping , Lymphocytes/cytology
15.
Nucleic Acids Res ; 18(21): 6319-24, 1990 Nov 11.
Article in English | MEDLINE | ID: mdl-2243778

ABSTRACT

Two new diphasmid vectors (lambda SK17 and SK22) and a novel procedure to construct linking libraries are described. A partial filling-in reaction provides counter-selection against false linking clones in the library, and obviates the need for supF selection. The diphasmid vectors, in combination with the novel selection procedure, have been used to construct a chromosome 3 specific NotI linking library from a human chromosome 3/mouse microcell hybrid cell line. The application of the new vectors and the strong biochemical and biological selections resulted in a library of 60,000 NotI linking clones. As practically all of them are real NotI linking clones (no false recombinants) the library represents approximately 3,000 human recombinants (equal to 10-15 genomic equivalents of chromosome 3). Previously published methods for construction of linking libraries are compared with the procedure described in the present paper. The advantages of the new vectors and the novel protocol are discussed.


Subject(s)
Chromosomes, Human, Pair 3 , Genetic Linkage , Genomic Library , Animals , Blotting, Southern , Cell Line , Cloning, Molecular/methods , DNA/genetics , DNA/isolation & purification , Deoxyribonucleases, Type II Site-Specific , Humans , Hybrid Cells/cytology , Karyotyping , Metaphase , Mice , Restriction Mapping
16.
Oncogene ; 5(8): 1207-11, 1990 Aug.
Article in English | MEDLINE | ID: mdl-2392324

ABSTRACT

Renal Cell Carcinoma (RCC) has been associated with the loss of heterozygosity at several loci on the short arm of chromosome 3 (3p). We have previously found that one of these loci, D3F15S2 (pH3H2) was lost in 76% of the tumor cells derived from heterozygous donors (Kovacs, G., Erlandsson, R., Boldog, F., Ingvarrsson, S., Müller-Brechlin, R., Klein, G. & Sümegi, J. (1988), Proc. Natl. Acad. Sci., 85, 1571-5). More recently we have identified a putative CpG island in the vicinity of D3F15S2, suggesting that DNA sequences in or around this site may have coding potential (Boldog, F., Erlandsson, R., Klein, G. & Sümegi, J. (1989). Cancer Genet. Cytogenet., 42, 295-306). The screening of a human placenta cDNA library with DNA probes derived from D3F15S2 has led to the isolation of several cDNA clones. They identified a 2.9 Kb long message in human placenta and kidney. In total RNA from 11 of 15 primary RCCs the gene expression was reduced to less than 20% compared to eight normal kidneys. This low level of expression may be due to contaminating normal tissue. In the remaining 4 tumors the expression varied from 24-51% compared to normal kidney. To facilitate reference, the gene was provisionally designated as 'RIK'. It was expressed in the HEK 293, one osteosarcoma (HOS), two carcinoma (COLO320 and QDMT), and three Burkitt lymphoma lines (BL2, BL29 and BL31). It was not expressed in one Burkitt lymphoma (DG75) and two EBV transformed lymphoblastoid cell lines (LCL) (NAD-20 and Cherry).


Subject(s)
Carcinoma, Renal Cell/genetics , Chromosomes, Human, Pair 3 , Kidney Neoplasms/genetics , Kidney/metabolism , Chromosome Aberrations , Chromosome Mapping , DNA/analysis , DNA/isolation & purification , Gene Expression , Heterozygote , Humans , Transcription, Genetic
17.
Acta Med Hung ; 41(4): 213-22, 1984.
Article in English | MEDLINE | ID: mdl-6240018

ABSTRACT

The episodic fluctuation of serum testosterone, androstenedione, dehydroepiandrosterone (DHA), dehydroepiandrosterone sulphate (DHAS) and cortisol levels was analysed to determine the reliability of a single blood estimate in characterizing the mean value of a 4 h period. Radioimmunoassay of the steroids was performed in sera of 8 apparently healthy women in various phases of the menstrual cycle in 10 min intervals between 08 and 12 h a.m. Reliability criteria of the methods used were comparable to those in common use. The within-person fluctuation of individual values was determined by the coefficient of variation of single estimates, and the methodological error of the single estimates was characterized with the maximum increment from the mean at 95% confidential limits. A maximum deviation of 30.8, 28.6, 37.0, 25.0 and 61.2%, respectively, found in the above order of steroids, suggests several hormone estimations to be necessary for judging hormone availability in a subject.


Subject(s)
Androstenedione/blood , Dehydroepiandrosterone/blood , Hydrocortisone/blood , Testosterone/blood , Adult , Analysis of Variance , Androgens/metabolism , Dehydroepiandrosterone/analogs & derivatives , Dehydroepiandrosterone Sulfate , Female , Humans , Menstrual Cycle , Time Factors
18.
Acta Med Hung ; 41(4): 203-11, 1984.
Article in English | MEDLINE | ID: mdl-6514478

ABSTRACT

The episodic fluctuation of serum progesterone, oestradiol and oestrone levels was studied. Progesterone was determined in the luteal phase, oestradiol and oestrone concentrations were measured in the proliferative and luteal phases in 8 subjects in 10 min intervals between 08 and 12 h a.m. Reliability criteria proved the radioimmunological methods used to be comparable to those employed routinely. The within-person fluctuation of hormone levels characterized by the coefficient of variation of single estimates averaged 20.16, 23.0 and 11.6% for progesterone, oestradiol and oestrone, respectively. The considerable fluctuation of hormone concentrations suggest the importance of hormone measurements to control luteal function and folliculogenesis during the menstrual cycle.


Subject(s)
Estradiol/blood , Estrone/blood , Progesterone/blood , Adult , Analysis of Variance , Estradiol/metabolism , Estrone/metabolism , Female , Humans , Menstrual Cycle , Progesterone/metabolism , Time Factors
19.
Acta Med Hung ; 41(4): 195-202, 1984.
Article in English | MEDLINE | ID: mdl-6440116

ABSTRACT

The episode fluctuation of serum LH, FSH and prolactin levels in peripheral blood was studied to determine the reliability of single estimates of the average blood level. Radioimmunoassay of the hormones was performed in serum withdrawn every 10 min between 8 and 12 h a.m. from 8 healthy women in various periods of the menstrual cycle. The rate of fluctuation of the hormone levels was characterized by the within-person coefficient of variation of single estimates, which averaged 30.3, 15.1 and 19.2% for LH, FSH and prolactin, respectively. Due to a pronounced fluctuation of gonadotropin levels, analysis of several serum samples is recommended to approach the actual mean hormone level.


Subject(s)
Follicle Stimulating Hormone/blood , Luteinizing Hormone/blood , Prolactin/blood , Adult , Analysis of Variance , Female , Follicle Stimulating Hormone/metabolism , Humans , Luteinizing Hormone/metabolism , Prolactin/metabolism , Time Factors
20.
Acta Physiol Acad Sci Hung ; 59(4): 349-58, 1982.
Article in English | MEDLINE | ID: mdl-7170987

ABSTRACT

This paper reports on partial purification and characterization of a natural (endogenous) factor capable of inhibiting in vivo DNA synthesis in mouse pneumocytes in a tissue-specific manner. By using a combination of ultrafiltration and various chromatographic techniques, the active agent has been partially purified from aqueous extracts of both bovine and rat pulmonary tissue. The factor responsible for the observed effect was found to be a heat labile compound, most likely a protein of approx. 40 000 molecular weight. Its chemical and physicochemical properties determined so far, and also the manner of its biological action implies that this lung tissue derived agent might be an endogenous proliferation inhibitor with a chalone-like character operating in the pulmonary epithelial cells.


Subject(s)
DNA/biosynthesis , Membrane Proteins/isolation & purification , Pulmonary Alveoli/ultrastructure , Tissue Extracts/isolation & purification , Animals , Cattle , Cell Membrane/analysis , Cell Membrane/metabolism , Chemical Fractionation/methods , Chromatography, Gel , Chromatography, Ion Exchange , DNA/antagonists & inhibitors , Depression, Chemical , Mice , Tissue Extracts/pharmacology , Ultrafiltration/methods
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