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PubMed1995

Effects of calcitriol on proliferation and differentiation of human fetal jejunum.

Several lines of evidence suggest a role for calcitriol in the regulatory mechanisms of intestinal development. Using organ culture as a model, we have verified whether this hormone influenced the cellular proliferation and differentiation processes of the developing human jejunum. We show that calcitriol supports a proliferative process in the fetal human jejunum at 11 weeks of gestation and a differentiating activity in samples obtained between 16 and 20 weeks. The present data therefore suggest that calcitriol has a biphasic effect on the development of the human intestinal epithelium and warrants caution in extrapolating to human, data observed in developing rodents and human carcinoma cell lines.

Calcitriol↗
PubMed2004

Expression and the biological activities of insulin-like growth factor-binding protein related protein 1 in rat uterus during the periimplantation period.

IGF binding protein-related protein 1 (IGFBP-rP1) is highly expressed in the rat uterus around the time of implantation. In the present study, we determined the periimplantation localization of IGFBP-rP1 mRNA and assessed the effects of recombinant IGFBP-rP1 on the proliferative and prostacyclin (PGI(2))-producing abilities of cultured endometrial cells early in pregnancy. IGFBP-rP1 mRNA was detected at high levels in endometrial stromal cells close to the smooth muscle of interimplantation sites around the time of implantation but absent from decidual zones surrounding the embryo. Differential uterine IGFBP-rP1 expression was also recognized in the delayed implanting pregnant model, but the level of mRNA decreased as decidual tissues formed in the decidualization model. Recombinant IGFBP-rP1 inhibited the proliferation of endometrial stromal cells in vitro and arrested them in the G(1) phase of the cell cycle. Furthermore, IGFBP-rP1 significantly stimulated PGI(2) synthesis and cyclooxygenase II mRNA expression in myometrial cells, both of which are essential molecules for successful implantation. These data suggest that IGFBP-rP1 is an implantation-associated protein and that it modulates the proliferation of rat uterine cells and their production of PGI(2) during the periimplantation period.

Animals↗
PubMed1999

Roles for PDGF-A and sonic hedgehog in development of mesenchymal components of the hair follicle.

Skin appendages, such as hair, develop as a result of complex reciprocal signaling between epithelial and mesenchymal cells. These interactions are not well understood at the molecular level. Platelet-derived growth factor-A (PDGF-A) is expressed in the developing epidermis and hair follicle epithelium, and its receptor PDGF-Ralpha is expressed in associated mesenchymal structures. Here we have characterized the skin and hair phenotypes of mice carrying a null mutation in the PDGF-A gene. Postnatal PDGF-A-/- mice developed thinner dermis, misshapen hair follicles, smaller dermal papillae, abnormal dermal sheaths and thinner hair, compared with wild-type siblings. BrdU labeling showed reduced cell proliferation in the dermis and in the dermal sheaths of PDGF-A-/- skin. PDGF-A-/- skin transplantation to nude mice led to abnormal hair formation, reproducing some of the features of the skin phenotype of PDGF-A-/- mice. Taken together, expression patterns and mutant phenotypes suggest that epidermal PDGF-A has a role in stimulating the proliferation of dermal mesenchymal cells that may contribute to the formation of dermal papillae, mesenchymal sheaths and dermal fibroblasts. Finally, we show that sonic hedgehog (shh)-/- mouse embryos have disrupted formation of dermal papillae. Such embryos fail to form pre-papilla aggregates of postmitotic PDGF-Ralpha-positive cells, suggesting that shh has a critical role in the assembly of the dermal papilla.

Actins↗
PubMed2001

Effect of diethylstilbestrol on cell proliferation and expression of epidermal growth factor in the developing female rat reproductive tract.

To evaluate mechanisms of cell proliferation in the fetal female rat reproductive tract, diethylstilbestrol (DES) effects on cell division and estrogen receptor (ER), epidermal growth factor (EGF) and EGF receptor (EGF-R) expressions were determined from gestational day (GD) 15.5 to 21.5. Reproductive tracts were evaluated within three regions along the Müllerian duct axis; these were proximal, middle and caudal, which differentiate into oviduct, uterus and upper vagina respectively. In fetuses from non-treated dams, epithelial and mesenchymal proliferation, as evaluated by 5-bromo-2'-deoxyuridine incorporation, was decreased with development in all regions of the Müllerian duct. EGF levels were determined by immunohistochemistry. Müllerian epithelial EGF immunoreactivity was intense in the proximal and middle regions on GDs 15.5 and 17.5. EGF staining remained intense only in the proximal epithelia by GD 19.5 and was weak in the caudal epithelium, but substantially reduced throughout epithelia in all regions by GD 21.5. Thus, decreased cell proliferation correlated with decreased EGF expression in the developing Müllerian duct. DES (100 microg/kg body weight) was injected from GD 15 to 19 and female fetuses were collected on GD 19.5. DES increased Müllerian duct cell proliferation in the proximal epithelium and mesenchyme but decreased it in the caudal epithelium compared with oil-treated controls. No proliferative DES effect was observed in any cell type in the middle region. Müllerian duct EGF immunoreactivity was suppressed by DES compared with oil. Competitive RT-PCR indicated DES also decreased mRNAs for EGF, ERbeta1 and ERbeta2, but not ERalpha and EGF-R. These results indicate EGF may be an important regulatory factor of Müllerian duct cell proliferation, and that DES may alter cell proliferation by disrupting normal EGF, ERbeta1 and ERbeta2 expression in the developing female rat reproductive tract.

Animals↗
PubMed1991

Chronic heat stress and prenatal development in sheep: II. Placental cellularity and metabolism.

Aspects of placental protein and energy metabolism were examined in pregnant ewes subjected to either thermoneutral (TN, 18 to 20 degrees C, 30% humidity, n = 7) or hot (H, 30 to 40 degrees C, 40% humidity, n = 5) temperatures through mid and late gestation. Fetal and placental weights and total content of protein, RNA, and DNA were reduced (P less than .001) in H ewes. Placental protein and RNA concentrations (mg/g) were not different, and DNA concentrations were slightly greater (P less than .1), in H vs TN ewes. Thus, heat seemed to greatly reduce total cell number and placentome size and only slightly decrease cell size. Ratios of RNA to DNA indicated a reduced capacity for protein synthesis in H placenta. However, in vitro fractional rates of protein synthesis in tissue slices from the fetal and maternal placenta and from the myoendometrium were not different between TN and H ewes. The H ewes had greater placental protein concentrations of hydroxyproline and glycine, perhaps suggesting a greater collagen content. In vitro oxygen consumption of fetal placenta, but not of maternal placenta or myoendometrium, was lower in H than in TN ewes. This lower oxygen consumption was partially due to a lower Na+,K+ ATPase-dependent oxygen consumption.

Animals↗
PubMed1997

Enhanced expression of cytokines may play a fundamental role in the mechanisms of immunologically mediated recurrent spontaneous abortion.

OBJECTIVES: To determine circulatory levels of IL-1 beta, TNF alpha, IFN-gamma and Neopterin in immunologically mediated recurrent abortion. DESIGN: Blood samples were withdrawn from both groups of first trimester recurrent abortion and controls for the determination of serum IL-1 beta, TNF alpha, IFN-gamma by the corresponding IRMA and serum neopterin by a double antibody RIA. SETTING: Department of Obstetrics & Gynecology, Cairo University Hospitals. PATIENTS: Thirty pregnant women in their first trimester who presented with inevitable abortion with history of at least 3 prior spontaneous consecutive abortions, in addition to twenty-three pregnant women in their first trimester presented with their first miscarriage due to chromosomal anomalies (controls). Every attempt was made to eliminate the possible known contributing factors of recurrent abortion except for the presence of cervical mucus sperm antibodies which were present in 24 out of 30 cases. INTERVENTIONS: None. RESULTS: Serum IL1 beta, TNF alpha and IFN-gamma levels of the abortion group were significantly elevated when compared to the corresponding levels of controls. The incidence of abnormal high values of these cytokines varied between 40 to 70%. There was no significant difference of serum neopterin between abortion and control groups. The increase of IFN gamma, the most active principle inducing neopterin release, was below the concentration needed for neopterin stimulation. CONCLUSIONS: In view of the proven findings that cytokines do have detrimental effects on implantation, trophoblast proliferation, embryo development and fetal survival, our results suggest a potential mechanism of immunologic recurrent abortion which involves the secretion of IL-1 beta, TNF alpha, IFN gamma and other cytokines by activated endometrial lymphocytes and macrophages in response to either trophoblast or sperm antigens and these cytokines could partially defuse or penetrate into the systemic circulation.

Abortion, Habitual↗
PubMed1995

[Clinical significance of proliferating cell nuclear antigen (PCNA) and argyrophilic nucleolar organizer region (AgNOR) in testicular tumors].

BACKGROUND: Proliferative potential of testicular tumor was assessed by immunohistochemistry using anti-proliferating cell nuclear antigen (PCNA) monoclonal antibody and silver staining of argyrophilic nucleolar organizer region (AgNOR), and the results were compared with clinical course. METHODS: Fourty-five patients with testicular tumor and 10 normal testicles were investigated. All specimens were fixed with 10% buffered formaliun within 24 hours. The interval between patient's recognition of testicular swelling for the first time and the date for operation was designated as M (months), and the specimen weight was designated as g (gram). Growth rate of the primary tumor was estimated roughly by g/M. RESULTS: PCNA positive rates in seminomas, non-seminomas and normal testes were 70.5 +/- 19.1% (mean +/- S.D.), 80.4 +/- 10.5% and 17.7 +/- 7.8%, respectively. PCNA positive rate in stage I seminoma (64.4 +/- 19.9%) was significantly lower than stage II and III seminoma (83.6 +/- 7.3%) (p < 0.05), but not significant between stage I non-seminoma and stage II, III non-seminoma. The mean numbers of AgNOR per nucleus in seminomas, non-seminomas and normal testes were 8.09 +/- 1.35 (mean +/- S.D.), 6.89 +/- 1.43, 4.18 +/- 1.60, respectively and significantly higher in testicular tumors than normal testes. There were, however, no significant difference between their clinical stages. Logarithmic significant correlation was observed between PCNA positive rate and the growth rate in primary lesion, but no correlation was found between mean number of AgNOR per nuclei and the growth rate. Of the 10 patients with stage I non-seminomas received surveillance policy, four relapsed. Of the 10 patients, 4 of the 6 patients with higher than 75% PCNA positive rate relapsed, whereas, none of the 4 patients with lower than 75% PCNA positive rate relapsed. There were no significant correlation between PCNA positive rate and number of AgNOR. CONCLUSION: PCNA is more accurate indicator of growth potential in testicular tumor than AgNOR and may predict recurrence in stage I non-seminomatous testicular tumor patients followed by surveillance policy.

Carcinoma, Embryonal↗
PubMed2000

Pleiotrophin can be rate-limiting for pancreatic cancer cell growth.

Pancreatic cancer is one of the most aggressive malignant tumors, with an overall survival rate of 2%. The identification of growth factors that contribute to the malignant phenotype can help to identify new targets for therapy. In this study, we analyzed the growth factor pleiotrophin (PTN) that was originally described as a developmentally regulated cytokine during early embryogenesis. More recently, PTN was found to be overexpressed in a variety of neuroectodermal tumors and described as an essential angiogenic growth factor in choriocarcinoma and melanoma, promoting metastatic growth. Recently, we discovered high expression levels of PTN in patients with gastrointestinal malignancies, particularly in those patients with pancreatic cancer. However, it is not known whether PTN is a contributor to the growth of pancreatic cancer or is only a bystander. We used ribozymes to deplete PTN mRNA from Colo357 pancreatic cancer cells and studied the resulting phenotype. The reduction of PTN resulted in a decrease in the proliferation rate, soft agar colony formation, and tumor growth in animals. Supplementation of cells with PTN partially reversed the ribozyme effect. The autocrine function of PTN was confirmed by using PTN-binding antibodies that inhibited the proliferation rate by 50% in Colo357 cells but also in a different pancreatic cancer cell line, Panc89. Our study identifies PTN as a new and essential growth factor for pancreatic cancer. Due to the restricted expression pattern of PTN in adults, PTN is suggested as a target for pancreatic cancer therapy.

Adenocarcinoma↗
PubMed2003

[Ultracytochemical localization of calcium and ATPase activity on the 2,4-D induced somatic embryogenesis of Lycium barbarum L].

In order to research the function mechanism of the 2,4-D during the development of plant somatic embryogenesis, we studied its function mechanism and relationship with the space-time distributing of Ca2+ content and ATPase activity on somatic embryogenesis of Lycium barbarum L. The possible effects on 2,4-dichlorophenoxyacetic acid (2,4-D) induced somatic embryogenesis and changes of Ca2+ and ATPase active at different development period of somatic embryogenesis. The result showed: The 2,4-D was a key hormone for induced embryonic state of Lycium barbarum L. The embryonic callus and non-embryonic callus was separately obtained in the medium that contains the auxin 2,4-D and lack 2,4-D. In the present study, we have observed the Ca2+ was more abundant in the further intercellular matrix and on the cell wall at the multi-cellular stage, and Ca2+ was concentrated in the plasma membrane and vacuoles membrane during embryonic cell differentiate and division, to the globular embryo, more Ca2+ was seen in the nucleus. Afterward, it was also observed to be distributed in the thicken cell wall and intercellular matrix. At the same process, the variations of ATPase activity and Ca2+ were highly similar, ATPase activity was mainly located on the plasma membrane in early embryogenic cells. With further development, it was also observed to be distributed in endoplasm, nucleus and vacuoles, with the thickening of embryogenic cell wall, ATPase activity was found in the thickened region and the intercellular space. However, the variations of ATPase activity and Ca2+ have not clearly observed variety dynamics at the nonembryogenic callus, and with further vacuolation of nonembryogenic cell, Ca2+ content and ATPase activity gradually drop. It was indicated there was a closely relationship between the dynamics of Ca2+ and ATPase activity in somatic embryogenesis by 2,4-D induced. And the space-time distribution of Ca2+ and ATPase activity play a key role on signal transmission and the regulation of relevant gene expression.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1983

Are there factors preventing cancer development during embryonic life?

On the basis of the following literature observations, a hypothesis is advanced that the development of cancer is actively inhibited during embryonic life. Although the processes of cell differentiation and proliferation are--without comparison--most pronounced during embryonic life, cancer is rarely found in the newborn and is seldom a cause of neonatal death or spontaneous abortion. Attempts to induce cancer in early-stage animal embryos by irradiation or by transplacental chemical carcinogenesis have been unsuccessful, even when exposed animals have been observed throughout their life-time. After the period of major organogenesis, however, the embryos become susceptible to carcinogenesis. In humans, the most common embryonic tumors arise in tissues which have an unusually late ongoing development and are still partly immature at or shortly before birth. For many human embryonic tumors the survival rates are higher, and spontaneous regression more frequent, in younger children, i.e. prognosis is age-dependent. Thus, although cancer generally appears in tissues capable of proliferation and differentiation, induction of malignancy in the developmentally most active tissues seems to be beset with difficulty. One possible explanation for this paradox could be that cancer is controlled by the regulators influencing development, regulators that are most active during embryonic life.

Animals↗
PubMedDate not supplied

[Division of regulatory cellular systems (Lvov)].

Two departments of the A. V. Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine were founded in 1969 in Lviv. These were: the Department of Biochemistry of Cell Differentiation headed by Professor S. I. Kusen and Department of Regulation of Cellular Synthesis of Low Molecular Weight Compounds headed by Professor G. M. Shavlovsky. The Lviv Division of the A. V. Palladin Institute of Biochemistry of the National Academy of Sciences of Ukraine with Professor S. I. Kusen as its chief, was founded in 1974 on the basis of these departments and the Laboratory of Modelling of Regulatory Cellular Systems headed by Professor M. P. Derkach. The above mentioned laboratory which was not the structural unit obtained the status of Structural Laboratory of Cellular Biophysics in 1982 and was headed by O. A. Goida, Candidate of biological sciences. From 1983 the Laboratory of Correcting Therapy of Malignant Tumors and Hemoblastoses at the Institute of Molecular Biology and Genetics, Academy of Sciences of Ukraine (Chief--S. V. Ivasivka, Candidate of medical sciences) was included in the structure of the Division. That Laboratory was soon transformed into the Department of Carbohydrate Metabolism Regulation headed by Professor I. D. Holovatsky. In 1988 this Department was renamed into the Department of Glycoprotein Biochemistry and headed by M. D. Lutsik, Doctor of biological sciences. In 1982 one more Laboratory of Biochemical Genetics was founded at the Department of Regulation of Cellular Synthesis of Low Molecular Weight Compounds, in 1988 it was transformed into the Department of Biochemical Genetics (Chief--Professor A. A. Sibirny). In 1989 the Laboratory of Anion Transport was taken from A. V. Palladin Institute of Biochemistry, Academy of Sciences of Ukraine to Lviv Division of this Institute. This laboratory was headed by Professor M. M. Veliky. One more reorganization in the Division structure took place in 1994. The Department of Glycoprotein Biochemistry (Chief--M. D. Lutsik, Doctor of biological sciences) was renamed the laboratory and a new Department of Regulation of Cellular Proliferation headed by R. S. Stoika, Doctor of biological sciences, was founded. Since 1992 the Division has its present name and exists as an independent research institution with the rights of the Institute of the National Academy of Sciences of Ukraine. At present about 100 researchers work at the Division, among them there are 8 Doctors and 27 Candidates of biological sciences. For the period of the Division existence 6 doctor's theses and 30 candidate theses were defended.(ABSTRACT TRUNCATED AT 400 WORDS)

Academies and Institutes↗
PubMed1995

Analysis of cell proliferation kinetics during the secondary palate development in quail.

A study was undertaken to analyze the spatio-temporal pattern of mesenchymal cell proliferation in the developing palate of quail. Quail embryos were grown in shell-less culture. The developing palates were labelled with 3H-thymidine between culture days 2-6 (which corresponded in vivo incubation days 5-9), and processed for light microscopic autoradiography. Percent labelled mesenchymal cells were determined. The data showed that, as in mammals, a high rate of random cell proliferation in mesenchyme was a major component of early palate development in quail. As the palate morphogenesis advanced, the rate of cell proliferation declined. Segmental analysis, however, indicated that, in contrast to mammals, the mesenchymal cell proliferation rates continually changed in various regions of quail palate during morphogenesis. It was suggested that the spatio-temporal changes in the distribution of dividing cells may reflect differences in the timings of cell cycles between various segments, thus resulting in a heterogeneous population of cells in the developing palate of quail. Further, the differences in the segmental pattern of cell proliferation between birds and mammals may form the basis for differences in the morphogenesis of their palates.

Animals↗
PubMed1994

3-Nitropropionic acid toxicity in cultured murine embryonal carcinoma cells.

The morphologic and histochemical effects of 3-nitropropionic acid (NPA) were examined in cultured murine embryonal carcinoma cells. NPA caused a dose-dependent inhibition of cell proliferation of cultured murine embryonal carcinoma cells at concentrations above 1.05 mM and was lethal at 4.2 mM. Morphologic changes included gross swelling of the cells, swelling of mitochondria and accumulation of organellar debris within the cytoplasm. NPA inhibited the activity of succinate dehydrogenase but not of malate, isocitrate or glucose-6-phosphate dehydrogenases, resulting in a decrease in intracellular ATP. Although succinate dehydrogenase activity was decreased by NPA, propionic acid and its mercapto-, 2-chloro-, and 3-chloro- derivates did not affect enzyme activity. 3-Nitropropanol also inhibited succinate dehydrogenase but only at a much higher concentration than was required with NPA. The results provide evidence that cytotoxicity caused by NPA results from inhibition of succinate dehydrogenase activity leading to depression of ATP synthesis. Loss of cellular integrity is probably a direct consequence of failure of energy-dependent cell homeostatic mechanisms such as the plasma membrane Na+/K+ pump, resulting in swelling and ultimately lysis of the cell.

Adenosine Triphosphate↗
PubMed1995

Gap junctions in brain glial cells and development.

In mammalian brain, glia are likely the most abundant and widespread cell population connected by gap junctions. Here, we summarize recent findings concerning the distribution, expression, and regulation of gap junction proteins in different glial subtypes and their possible roles in modulating the function of the nervous system. Intercellular communication through gap junctions is proposed to be one of the required interactions between cells during development, and we discuss how gap junctions could participate in development of brain glial cells.

Aging↗
PubMed1996

Intestinal growth in altricial European starling (Sturnus vulgaris) and precocial Japanese quail (Coturnix coturnix japonica). A morphometric and cytokinetic study.

This paper introduces the comparison of altricial and precocial birds as a model system for the study of patterns of intestinal growth. It reports on comparative morphometric and cytokinetic studies of intestinal growth in altricial European starling (Sturnus vulgaris) and precocial Japanese quail (Coturnix coturnix japonica). Morphometric data at five gut segments of neonates and adults of both species are provided to evaluate the growth of the intestine. A double labelling technique combining 3H-thymidine autoradiography with 5-bromo-2'-deoxy-uridine immunohistochemistry gives labelling index, length of S-phase, and tissue turnover time for hatchlings and adults of both species. The results of morphometry show a similar pattern of intestinal growth in both species, a strong dependency of intestinal growth on body size, and a clear topographic separation of areas of cell proliferation from the functional mucosal epithelium. The results of the cytokinetic experiments indicate a constant length of the S-phase independent of species, development, and age. The 3H-thymidine labelling index is significantly higher in neonate quail than in neonate starling. Between adult animals of both species, the labelling index goes not differ significantly except for the duodenum which has a higher index in starling. The turnover time necessary to replace the mucosal epithelium is 3 days in hatchlings of both species and ranges between 10 and 17 days in adults. A diurnal pattern of cell proliferation, as previously reported in adult quail, is not present in neonate quail. Thus, differences in intestinal growth among altricial and precocial birds are primarily based on size differences of proliferation areas whilst cellular proliferation rates have only minor impact on intestinal growth. The short turnover time and the high rates of proliferation suggest that the intestine is a flexible organ that may be adjusted in size and function to the actual needs of the growing bird.

Aging↗
PubMed1997

Expression of CD30 and CD30 ligand in cultured cell lines from human germ-cell tumors.

Embryonal carcinoma (EC) cells are the multipotent stem cells of human teratocarcinomas. EC cell lines isolated from human testicular teratocarcinomas retain pluripotentiality and show cell-surface marker phenotype and growth characteristics similar to those of primate embryonic stem cells. CD30, a member of the tumor necrosis factor superfamily of cytokine receptors, is expressed on the surface of human EC cells, but little is known regarding its function or the expression of its ligand in this context. Northern blot analysis, reverse transcriptase-PCR, and immunoprecipitation were used to study the expression of CD30 ligand and CD30 in cell lines representing multipotent and nullipotent EC as well as yolk sac carcinoma. Antibodies, recombinant CD30-Fc, and recombinant CD30 ligand were used in bioassays to assess the ability of exogenous CD30 ligand to promote the growth of multipotent human EC stem cells. Nullipotent and multipotent human EC cells, but not yolk sac carcinoma, expressed surface CD30, as demonstrated by immunoblotting and immunofluorescence; in multipotent cells, expression was decreased during differentiation induced by retinoic acid. Transcripts for CD30L were found at high levels in a yolk sac carcinoma cell line by Northern analysis and reverse transcriptase-PCR and were present at lower levels in EC cells as well. Immunoprecipitation confirmed expression of CD30 ligand protein in yolk sac carcinoma and in nullipotent embryonal carcinoma. Antibodies with CD30 cross-linking activity failed to promote the growth of multipotent EC cell line GCT 27 X-1. Anti-CD30 ligand antibodies and CD30-Fc both failed to block the promotion of EC cell growth induced by yolk-sac cell culture supernatants or feeder-cell monolayers. CD30 ligand is expressed in cell lines representative of human yolk sac carcinoma and nullipotent embryonal carcinoma. The membrane-bound cytokine may have an autocrine role in human EC stem-cell renewal, but other exogenous factors are required to maintain the growth of multipotent EC cells in vitro.

3T3 Cells↗