Biomedical subjects

G L Engelmann

Publications and source records attributed to G L Engelmann.

30 records · Page 2Linked to original sources

PubMed1989

Retroviral v-myc infection of primary fetal liver cells: transformation of monocytes in vitro.

Oncogenes carried by retroviruses can alter the growth properties of many cell types. We examined the molecular mechanism by which a retrovirus containing one or a combination of oncogenes can transform and immortalize hematopoietic cells. Murine fetal liver cells were used as an enriched source of early hematopoietic cell progenitors; the cells were infected with a series of recombinant murine retroviruses capable of expressing the avian v-myc, v-H-ras and v-raf oncogenes. Three factor-independent cell lines were obtained: FL-ras/myc, FL-J2 (v-raf/v-myc) and FL-myc, a unique cell line generated using a single oncogene. Cytochemical, morphologic and phenotypic analyses indicated that these cell lines were of the monocyte lineage. Southern and Northern blot analyses revealed that the three cell lines had integrated viral DNA and were expressing the mRNA transcripts corresponding to these viral oncogenes. To examine the mechanism of factor independence, supernatants from these cell lines were tested for CSF-1 activity. Supernatants from FL-myc and FL-ras/myc cells were shown to contain CSF-1 activity and Northern blot analysis of the three cell lines revealed the presence of mRNA transcripts for the CSF-1 and c-fms genes. It is possible that the growth factor independence of these cell lines is related to the development of autocrine-induced proliferation.

Animals↗

PubMed1988

Biochemical characterization of neonatal cardiomyocyte development in normotensive and hypertensive rats.

Final cellular replication and functional maturation of the mammalian ventricular myocyte (cardiomyocyte) is known to occur during the first 2 post-natal weeks in rodents, yet the exact temporal sequence of events remains to be clearly established. When a genetic predisposition toward ventricular hypertrophy also manifests its influence during this time period, possible growth aberrations may occur. Using the spontaneously hypertensive rat (SHR) as an animal model, we have determined that biochemical and morphological indices of cardiomyocyte replicative deficits occur during the first 5 to 8 post-natal days relative to the normotensive control strain, Wistar Kyoto (WKY). Reduced ventricular nucleic acid (DNA and RNA) content and disproportionate RNA- and protein-DNA ratios were found in the neonatal SHR. Incorporation of tritiated thymidine into acid precipitable DNA during the first 5 postnatal days was also reduced in the SHR. Morphological evidence of reduced thymidine incorporation was determined autoradiographically at the cellular and nuclear level using isolated cells and nuclei, respectively. The results of our study indicate that SHR cardiomyocyte hyperplasia is reduced during the first 5 to 8 post-natal days when the last "window" of ventricular myocyte cellular replication occurs. Therefore, the maturing ventricle of the neonatal SHR contains fewer cardiomyocytes than its normotensive control and these cells appear to enter a hypertrophic growth phase at an accelerated rate.

Animals↗

PubMed1988

Insulin-like growth factor-I messenger ribonucleic acid in the developing human placenta and in term placenta of diabetics.

Fetal growth and development are dependent upon the growth and development of the placenta. Control of placental growth and development is little understood. Immunoreactive insulin-like growth factor-I and -II (IGF-I and IGF-II) have been shown to be released by human placental tissue and human placental membranes have been observed to contain specific receptors for these growth factors. Furthermore, we have demonstrated the presence of IGF-II mRNA transcripts in the developing human placenta and at gestational term in placentae of diabetics. Thus, the IGFs may have a regulatory role in the growth and development of the placenta via autocrine and/or paracrine mechanism(s) of action. In this report we demonstrate the presence of four differing size species of placental poly(A)+ RNA which specifically hybridize to an IGF-I probe originally isolated from an adult human liver cDNA library and localize IGF-I and IGF-II mRNA to syncytiotrophoblasts and fibroblasts, respectively, of the placenta by in situ hybridization. The major transcript is 7500 bases in size and the remaining three transcripts are 5000, 1100, and 900 bases in length with no apparent changes from these sizes throughout gestation and at term in diabetics. Quantification by densitometry of placental IGF-I mRNA detected by dot blot hybridization indicated that first and second trimester placentae each express more IGF-I mRNA relative to that expressed in placenta at term. These results suggest that there are developmental changes in the relative amount of IGF-I mRNA expressed in the human placenta. IGF-I is, therefore, most likely important early in gestation as a placental growth factor. This time period is critical for fetal development and growth, when embryonic induction, organogenesis, and rapid cell proliferation occur.

Female↗

PubMed1988

Removal of surgically induced fibrous arterial plaques by argon ion laser angiosurgery using a multifiber delivery system. An experimental study in the dog.

Removal of intravascular atherosclerotic obstructions by laser irradiation has gained the attention of many investigators, but has proven to be considerably more difficult to accomplish than initially envisioned. We tested, in an animal model, an argon ion laser delivery system that permits control of (1) laser power, (2) exposure time, and (3) laser beam spot size. The study was conducted on surgically, induced focal fibrous plaques in the carotid arteries of nine dogs. Plaque removal, vessel patency, and healing were evaluated angiographically and by light and electron microscopy at intervals up to 60 days after treatment. Results showed that intravascular obstructions could be removed, healing occurred, and vessels remained patent for up to 60 days.

Animals↗

PubMed1987

Effect of sodium butyrate on primary cultures of adult rat hepatocytes.

Sodium butyrate, at millimolar concentrations, seems to mediate or initiate multiple effects on many mammalian cells in culture. Although many transformed cell lines respond to butyrate treatment with acquisition of normal cellular characteristics, the effect of butyrate on a normal cell type, the parenchymal hepatocyte, has not been studied. Serum-free primary cultures of adult rat hepatocytes maintain many adult characteristics, yet after several days in culture a loss of adult characteristics occurs while fetal characteristics are often reexpressed. Therefore, we investigated whether butyrate treatment would improve the morphologic and biochemical characteristics of cultured hepatocytes. Exposure to 5 mM butyrate for 3 d did not affect hepatocyte viability or morphology but retarded the progressive decline in cytochrome P-450 levels and 5'-nucleotidase activity. The spontaneous increase in alkaline phosphatase activity was reduced and the induction of tyrosine aminotransferase was inhibited after 3 d in culture. The fetal liver characteristic, gamma glutamyltranspeptidase, was not affected by butyrate treatment. Results of this study suggest that butyrate represents a nontoxic compound capable of improving the maintenance of cell culture characteristics of adult rat hepatocytes.

5'-Nucleotidase↗

PubMed1987

Morphometric analysis of cardiac hypertrophy during development, maturation, and senescence in spontaneously hypertensive rats.

Hypertrophy of the mammalian heart, regardless of the initiating event, results in architectural remodeling of ventricular components that maintain structural and functional characteristics of this organ. Ventricular components that vary their morphology and morphometry in a hypertrophic state are the muscle cells, connective tissue elements, vasculature, or a combination of some or all of the above. Morphologic quantification of the progressive tissue changes occurring throughout the natural life span of the spontaneously hypertensive and normotensive Wistar-Kyoto rats has not been thoroughly documented. Using perfused-fixed tissue from both strains at 1, 6, 12, 18, and 24 months of age, we have determined the morphometric changes that occurred in the subepicardial and midwall regions of the left ventricle. Myocyte cell size, wall thickness, and arterial blood pressure were elevated in 1-month-old spontaneously hypertensive rats, reached significance by 6 months, and remained significantly greater throughout the 24 months examined. Tissue morphometry demonstrated significant tissue component volumetric differences at 6 months in the spontaneously hypertensive rat. Age-related morphometric tissue changes occurred in both strains yet were exacerbated (percent volume of myocytes) or diminished (percent volume interstitial space) in the mature and aging spontaneously hypertensive rat. Capillary density of SHR left ventricle showed a drastic decline so that 6-month-old SHR had the same density as a senescent Wistar-Kyoto. Tissue morphometry and capillary density data strongly support the hypothesis that tissue oxygenation is diminished in the spontaneously hypertensive rat, and as a result, tissue necrosis and myocyte cell death occur.

Aging↗

PubMed1986

Effects of levamisole on primary cultures of adult rat hepatocytes.

Levamisole represents one of several new compounds that exhibit immunomodulating activity. Pharmacological data have documented a relationship between liver drug metabolism of levamisole and its subsequent immunomodulating activity. To directly investigate this relationship in a controlled manner, primary cultures of adult rat hepatocytes were treated with levamisole, and ultrastructural and biochemical effects were analyzed. Ultrastructurally, levamisole did not disrupt the cellular architecture of the hepatocytes. Biochemically, levamisole stimulated alkaline phosphatase activity and elevated microsomal cytochrome P-450 content after a 48-hr incubation. High pressure liquid chromatographic analysis of levamisole metabolites produced by cultured hepatocytes suggested the formation of a hepatocyte-specific metabolite(s) that may be associated with its immunological mode of action.

5'-Nucleotidase↗

PubMed1986

Age-related changes in ploidy levels and biochemical parameters in cardiac myocytes isolated from spontaneously hypertensive rats.

Postnatal growth of the mammalian ventricular myocyte is characterized by a brief period of hyperplasia followed by an extensive period of physiological hypertrophy. Using myocytes isolated from both Wistar-Kyoto and spontaneously hypertensive rats from fetus to 10 months old, we analyzed morphological, biochemical, and ploidy changes. Fetal myocytes from both spontaneously hypertensive and Wistar-Kyoto rats were mononuclear, diploid cells. By 4 weeks, adult binucleation levels (84% binuclear) were found, and myocytes pooled from both ventricles demonstrated nuclear ploidy shifts to tetraploid levels. However, analysis of myocytes isolated from left ventricle, right ventricle, and septum showed that nuclear polyploidation was confined to the right ventricle and septum, with few polyploid nuclei detected in the left ventricle. This pattern remained relatively constant through 10 months of age, although spontaneously hypertensive myocytes showed significantly more polyploidation than Wistar-Kyoto in all regions. Biochemical analysis of isolated myocytes substantiated the nuclear ploidy changes and demonstrated elevated protein and ribonucleic acid content in left ventricular myocytes without extensive polyploidation. Since cardiac hypertrophy, both physiological and pathological, is associated primarily with the left ventricle and occurs in the absence of significant ploidy changes, these findings suggest that a unique pattern of gene regulation may be ongoing in the left ventricle myocytes that is not present in the septum and right ventricle. These variations may be essential for cellular hypertrophy under normal and pathological conditions.

Aging↗

PubMed1985

Maintenance and induction of cytochrome P-450 in cultured rat hepatocytes.

Maintenance of microsomal cytochrome P-450 content by cultured rat hepatocytes has proven an elusive goal. It is reported here that exogenous heme maintains cytochrome P-450 content of cultured rat hepatocytes at high levels during the first 72 h of incubation. The maintenance studies have been expanded to demonstrate the in vitro induction of cytochrome P-450 by phenobarbital treatment. The induction of P-450 in vitro by phenobarbital required the trace element, selenium, in the presence of exogenous heme. The present findings suggest that selenium, and other trace elements, may have an essential role in the formation of holocytochrome P-450 in vitro.

5-Aminolevulinate Synthetase↗

PubMed1981

Age-related changes in isolated rat hepatocytes. Comparison of size, morphology, binucleation, and protein content.

Hepatocytes isolated from 6-, 12-, 18-, and 30-month-old female Fischer F344 rats were examined by scanning electron microscopy. No significant change in cell size with age was observed. However, the surface morphology of the cells isolated from the older animals exhibited a significant increase in surface folds. This feature did not exceed 10% of the cell population until 12 months of age and continued to increase to 31% of the cells in 30-month-old rats. From 6 to 12 months of age, there was a significant increase in protein content of the hepatocytes. No further increase in protein content occurred during senescence. An increase in percentage of binuclear cells occurred after 24 months of age. Because ploidy and binucleation increase with increasing age, it appears that the nuclear/cytoplasmic ratio changes as a function of age.

Aging↗

PubMedDate not supplied

Expression of cardiac muscle markers in rat myocyte cell lines.

Recently developed rat heart myocyte cell lines have afforded us the opportunity to evaluate the expression of several transcription factors associated with early cardiac development. These factors include, but are not limited to, Nkx-2.5/Csx, MEF-2C and MLP (Muscle LIM Protein). These factors have been shown to be temporally expressed in pre-cardiac mesenchyme coincident with the earliest stages of heart development. Using the BWEM and CLEM myocyte cell lines as models of the embryonic, committed cardiomyocyte, we have evaluated the basal expression levels of these three genes over multiple passages. Both cell lines express these genes, with MEF-2C being the most abundant based on Northern blot hybridization analyses. Interestingly, as these cells increased their passage number, there was a corresponding increase in their basal expression levels. To evaluate potential 'downstream' effectors of these genes, we examined the basal expression levels of two cardiac-specific genes cTNC and MLC-2v. Transcript levels for both of these contractile filament genes were elevated with passage, suggestive of a inductive process mediated by one or all these three transcription factors. Promoter analysis of MLC-2v expression in the CLEM line shows that this increase is transcriptionally-mediated and the lines retain the necessary regulatory factors to maintain and control the transcription of this gene. Analysis of the dynamics of the regulatory role(s) that these three transcription factors play in cardiac development can now be evaluated in a homogeneous, cell culture system.

Animals↗

PubMedDate not supplied

Immediate postnatal rat heart development modified by abdominal aortic banding: analysis of gene expression.

Proliferative growth of the ventricular myocyte (cardiomyocyte) is primarily limited to embryonic, fetal and very early neonatal periods of heart development. In contrast, cardiomyocyte maturation, as evidenced by cellular hypertrophy, is a long-term process that can occupy the bulk of the life-span of the mature organism. As the newborn myocyte undergoes a 'transition' from proliferative to hypertrophic growth, ventricular remodeling of the non-myocyte compartment is characterized by increased extracellular matrix (ECM) formation and coronary capillary angiogenesis. A role for ventricular-derived growth factors (GFs) in these inter-related processes are examined in an animal model of altered heart development produced by neonatal aortic banding. The suprarenal abdominal aorta of five day old rat pups were banded (B), sham operated (S), or untreated (C) and ventricular tissue (left ventricular free wall and septum) obtained at 7-, 14-, and 21-days post-intervention. Using Northern blot RNA hybridizations, expression of growth factors (GFs) and/or GF-receptors (GFR's) temporally associated with heart development were evaluated. Transcript levels for TGF-beta 1, IGF-II, and their associated cell surface receptors were increased in B animals. Concomitant changes in extracellular matrix (ECM) genes (as evaluated by Collagens Type I, III, and IV) were also increased in B animals. In addition, transcript levels for the vascular morphogenesis and remodeling-related protein SPARC (Secreted Protein, Acidic and Rich in Cysteine) was also elevated in the B animals. In several instances, S animals demonstrated changes in steady state transcript levels for genes which may influence myocyte maturation during the postnatal period. This suggests that normal autocrine/paracrine growth regulatory stimuli and responses can be modified (by surgical intervention and/or abdominal aortic banding) and these perturbations in gene expression may be related to previously documented changes in myocyte cell number, vascular composition, and ventricular architecture of the banded, neonatal heart. Future studies using this model will provide an opportunity to evaluate and possibly identify the stimuli and signal transduction machinery that regulate the final phases of myocyte proliferation, stimulate capillary formation and ECM deposition, and orchestrate the transition to hypertrophic growth during heart development.

Animals↗