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Biomedical subjects

R L Noble

Publications and source records attributed to R L Noble.

At least 37 records · Page 2Linked to original sources
PubMed1983

Receptor-mediated mitogenic action of prolactin in a rat lymphoma cell line.

PRL and other lactogenic hormones are potent mitogens in a lymphoma cell line derived from a lymph node of an estrogenized Noble (Nb) rat. The present study demonstrates that these cells (designated Nb2 node) possess receptors that bind only lactogenic hormones. There are approximately 12,000 receptor sites per cell. The kinetics of binding of [125I]iodo-PRL exhibited by Nb2 lymphoma cells is unusual in that PRL binding follows a biphasic pattern. Binding of [125I]iodo-PRL reaches a maximum in 1 h at 37 C, followed by a rapid decline. This pattern was not observed if binding was carried out in the presence of chloroquine, a lysosomotropic agent, or if cell homogenate was used for binding. We also examined the relationship between receptor occupancy and the magnitude of PRL response in these cells. Maximal growth stimulation by PRL occurs when only 35% of the maximal binding of PRL is achieved, suggesting that a majority of the PRL binding sites may be spare receptors. This study also revealed that the dissociation constant (Kd) of the PRL receptors in Nb2 cells is 75 pM, approximately 20-fold higher than that of the receptors in other cell types. PRL at 6 pM produces a half-maximal growth response in the Nb2 cells. Antibodies against the PRL receptors are able to abolish the PRL-induced proliferation of Nb2 cells. In the absence of PRL, these antibodies alone can induce proliferation of Nb2 cells, mimicking the action of PRL. Divalent (Fab)2 fragments, but not monovalent Fab, were also effective. These observations suggest that antibodies to the receptor, or the hormone itself, initiate a biological response possibly by cross-linking PRL receptors on the cell membrane, and that the entry of the PRL molecule, or fragments of it, into the intracellular compartment is not necessary for the biological action of PRL.

Animals↗
PubMed1983

Influence of malnutrition and alterations in dietary protein on murine rotaviral disease.

The possible correlation between malnutrition and degree of severity of rotavirus-associated infantile diarrhea which appears to occur in human populations was studied using a mouse model. To determine the effects of general malnutrition or altered levels of dietary protein, female mice were fed throughout pregnancy and infection periods with diets diluted with 0, 300, or 600 g glucose/kg, designated as normal nutrient to calorie ratio (N/C) diet, 70% N/C diet, or 40% N/C diet or with diets containing 75, 150, or 300 g casein/kg, as low-, normal-, or high-protein diets. Murine rotavirus was given by gavage to the 2-day-old offspring of these dams, and the extent of infection determined. Marked increases in severity of diarrheal disease were seen in the infants from dams receiving the 40 and 70% N/C diets and the low-protein diet. Severity of infection was seen as increased deaths, reduced weight gain, and increased passage of diarrheic feces. Intestinal viral levels and intestinal diarrhea scores did not vary appreciably. Serum interferon remained below detectable limits throughout the studies, but serum antibody was determined in dams 30 days post-virus exposure. The latter titers were lower in the infected mice from dams fed the 40 and 70% N/C diets, but were essentially the same in all the protein diet groups. Cross-fostering was done using the 40 and 100% N/C diets, wherein mice from dams fed either diet were placed on mothers fed the opposite diet. Increased severity of infection was again seen when the virus was given 2 days after the exchange, although the greatest infection occurred in animals from dams fed 40% N/C diet which were then fostered by other similarly fed dams. The increased host sensitivity to the rotaviral infection appeared to be a result of both pre- and postnatal dietary effects.

Animals↗
PubMed1982

Effect of iodination on human growth hormone and prolactin: characterized by bioassay, radioimmunoassay, radioreceptor assay, and electrophoresis.

Human GH (hGH) and PRL (hPRL) were iodinated using lactoperoxidase. The iodinated hormones were characterized by RIA, radioreceptor assay (RRA), and bioassay (BA) using the Nb2 Node lymphoma cell line. The proportion of tracer that could bind to rat liver membranes or rabbit antibodies was determined, and the distribution of iodinated hormones was examined using polyacrylamide gel electrophoresis. Excess antibody was capable of precipitating 87.9% of the radioactivity associated with the hGH tracer and 86.0% of the hPRL tracer. The maximal specific binding to a liver membrane preparation averaged 67.3% of the [125I]iodo-hGH radioactivity and 48.8% of the [125I]iodo-hPRL radioactivity. The respective BA and RRA activity estimates for [125I]iodo-hGH averaged 90% and 114% of the activity measured by the RIA. For [125I]iodo-hPRL, the values were 75% by BA and 68% by RRA. The bioactivity profiles of iodinated hGH and hPRL shifted anodally on polyacrylamide gel electrophoresis in comparison to the bioactivity distribution of the respective uniodinated hormones. Iodine incorporation rather than oxidation appeared to be responsible for the shift. After electrophoresis, all eluates which contained significant radioactivity were active in the BA and RIA. Furthermore, specific activities calculated from the bioactive hormone and radioactivity in each electrophoretic segment agreed well with the average specific activity estimated from the amount of iodine incorporated into the protein peak upon gel filtration. These data suggest that hGH and hPRL to a major degree retain biological integrity after iodination.

Biological Assay↗
PubMed1982

Spontaneous rat nephroblastoma: ultrastructure of a transplant line.

Tumor tissue derived from a nephroblastoma arising spontaneously in an Nb hooded rat and carried for 56 generations as a subcutaneous syngeneic transplant was examined by electron microscopy. Histologically, the transplant showed the typical pattern of dense clusters or sheets of basophilic epithelioid cells set within ramifying tracts of fibrous mesenchyme. At the ultrastructural level, the tumor cells within dense clusters were arranged in anastomotic cords that circumscribed clear intervening spaces. The tumor cells were a monomorphic population possessing the morphologic characteristics of blast cells, including polyribosomes as the predominant cytoplasmic organelle. Furthermore, the close contiguity of cells within the cords, their polyhedral form, and their connection by intercellular junctions established the tumor cell type as epithelial. Consistent with benign stroma, mesenchymal tracts contained highly differentiated, mature fibroblasts, collagen, phagocytes, cells of the lymphoid series, and normal blood vessels. No transitional forms between epithelium and mesenchyme suggestive of bipotential differentiation were seen. Although the data represent only a single transplantation line, the observations support the contention (based on light microscopic appraisal of a number of primary tumors) that nephroblastoma in te rat can be a purely epithelial neoplasm.

Animals↗
PubMed1981

Comparison of the interactions of a specific neurotoxin (alpha-bungarotoxin) with the acetylcholine receptor in Torpedo californica and Electrophorus electricus membrane preparations.

alpha-Bungarotoxin, a snake neurotoxin, binds irreversibly and specifically to the acetylcholine receptor isolated from the electroplax of Electrophorus electricus and Torpedo species and has been an important tool in the study of the receptor-ligand binding mechanism. Two distinct kinetic processes have been observed in studies with membranes from E. electricus. A minimum mechanism for the toxin reaction involves (i) the reversible binding of two toxin molecules to the receptor prior to the irreversible formation of toxin receptor complexes and (ii) a toxin-induced conformational change of the receptor which leads to an increase in the affinity of the receptor binding sites for toxin [Hess, G. P., Bulger, J. E., Fu, J.-j. L., Hindy, E. F., & Silberstein, R. J. (1975) Biochem. Biophys. Res. Commun. 64, 1018-1027]. Only one process has been detected in Torpedo membranes. Here, we determine whether the receptors in Torpedo californica and E. electricus membranes have different properties or whether the measurements and their interpretation were responsible for the different results. Two methods which are frequently used in binding studies to separate free and bound toxin, a CM-52 cellulose minicolumn assay and DE-81 filter disk assay, have been compared. The results obtained indicate that the interaction of toxin with receptor from T. californica is similar to that observed with receptor from E. electricus. The apparent differences which have been reported in the literature are shown to have arisen from the design of the experiments in which T. californica membranes were used.

Animals↗
PubMed1981

Occurrence, transplantation, and histologic characteristics of nephroblastoma in the Nb hooded rat.

Ten kidney tumors conforming to nephroblastoma were encountered during a 10-year period in a closed colony of Nb hooded rats. Histologic examination of the primary neoplasms, and of a limited number of transplanted derivatives, indicates that spontaneous nephroblastoma in the rat is exclusively an epithelial neoplasm, consisting of dense aggregates of undifferentiated, basophilic blast cells set in a connective tissue stroma of benign character. This uniform entity should be clearly distinguished from renal mesenchymal tumor, a connective tissue neoplasm of heterogeneous form which is induced frequently in rats by certain chemical carcinogens.

Age Factors↗
PubMed1981

Progressive hyperplastic lesions of the bladder uroepithelium after hormone stimulation in Nb rats.

The bladder epithelium of female Nb rats was markedly stimulated by treatment with pellets of testosterone propionate. Estrone treatment simultaneously reduced the incidence of bladder changes to approximately 50 per cent although estrone alone had no effect on bladder weights. The bladders of male rats were less responsive to hormones and were stimulated only by treatment with estrogen + testosterone. Massive papillary hyperplasia of the uroepithelium that extended into the kidney pelvis could be produced by hormones in the absence of calculi. Transplants of hyperplastic bladder epithelium usually resulted in small pseudo-bladders, but in one case a transplantable squamous cell carcinoma developed.

Animals↗
PubMed1980

Nuclear binding of androgens and acid phosphatase activity in prostatic tumors of Nb rats.

A transplantable prostatic adenocarcinoma derived from the dorsal lobe of the prostate gland of an Nb rat was analyzed for the concentration of nuclear androgen-binding sites and the presence of acid phosphatase activity. When extracts of nuclei from normal prostatic tissue were labelled with [1,2-3H]dihydrotestosterone in the absence and presence of competitor, two types of specific binding were observed: type 1 was characterized by an association constant (Ka) of 6 x 10(7) M-1 and involved a molecule that was excluded from Sephadex G-200; type 2 was characterized by a Ka of 3 x 10(8) M-1 and depended on a binding component that was retained by Sephadex G-200. Nuclei from androgen-stimulated tumors contained reduced concentrations of both androgen-binding components, whereas nuclei from autonomous tumors had only a trace amount of type 1 sites and were entirely devoid of type 2 sites. In all tumors the acid phosphatase activity per mg of protein was markedly elevated. Relative to normal, the activity of this enzyme was 140% and 350% higher in androgen-stimulated and autonomous tumors, respectively. These findings indicate that prostatic tumors are characterized by a decrease in nuclear androgen-binding, and an increase in specific activity of acid phosphatase, and also that such changes are more pronounced in autonomous than in androgen-stimulated tumors.

Acid Phosphatase↗
PubMed1980

Plasminogen activator in prostatic tumors of Nb rats.

Using a fluorometric assay, nonspecific proteolytic activity and plasminogen activator were measured in transplantable tumors of the dorsal prostate of Nb rats. Nonspecific proteolytic activity in prostatic tumors did not differ significantly from that measured in normal dorsal prostate, whereas plasminogen activator activity, undetectable in the latter tissue, was readily measurable in the tumors. Furthermore, plasminogen activator activity in prostatic tumors characterized by hormone-insensitive growth was 8-fold higher than in tumors characterized by androgen-stimulated growth. In both typess of tumors, the plasminogen activator activity per mg protein was highest in the lysosomal fractions. The results indicate that plasminogen activator may be a useful marker for discriminating between androgen-stimulated and autonomous prostatic tumors.

Adenocarcinoma↗
PubMed1980

Inactivation (desensitization) of the acetylcholine receptor in Electrophorus electricus membrane vesicles by carbamylcholine: comparison between ion flux and alpha-bungarotoxin binding.

The inactivation (desensitization) of the acetylcholine receptor by carbamylcholine, a stable analogue of acetylcholine, has been investigated in eel Ringer's solution, pH 7.0, 0 degrees C, by measurements of (i) ion flux and (ii) the kinetics of the reaction of [125I]-alpha-bungarotoxin with the receptor. The effect of preincubation with carbamylcholine is significantly different in the two types of measurement. In both the receptor-controlled flux of inorganic ions and the toxin-binding kinetics a biphasic process has been observed (Hess, G.P., Lipkowitz, S., Struve, G.E., 1978, Proc. Nat. Acad. Sci. USA 75:1703; Hess, G.P. et al., 1975, Biochem. Biophys. Res. Commun. 64:1018; Bulger, J.E. et al., 1977, Biochemistry 16:684), only the initial fast phase of which is inhibited and the subsequent slow phase persists. However, preincubation with carbamylcholine per se has no effect on the toxin reaction. The results obtained are consistent with the proposal of Katz and Thesleff (Katz, B., Thesleff, S., 1957, J. Physiol. (London) 138:65) that the active form of the receptor is converted to an inactive form in the presence of acetylcholine receptor ligands, and with our previous experiments (Hess et al., 1978) which indicated that one receptor form is responsible for the initial fast phase of both the receptor-controlled ion flux and the toxin binding reaction, and that its conversion to the other form results in the slow phases in these two measurements.

Animals↗
PubMed1980

A new sensitive and specific bioassay for lactogenic hormones: measurement of prolactin and growth hormone in human serum.

The replication of Nb 2 Node rat lymphoma cells in suspension culture is specifically stimulated by lactogenic hormones. Human (hPRL), ovine, bovine, and rat PRLs stimulated replication in a dose-dependent manner in the concentration range of 10 pg/ml to 1 ng/ml. Human, ovine, and bovine placental lactogens were similarly active. In addition, cell replication was stimulated by human GH (hGH), which is known to have lactogenic activity. Other hormones and growth factors examined were inactive. The growth stimulatory effects of hPRL and hGH were completely inhibited when excess anti-hPRL and anti-hGH, respectively, were added to the medium. A bioassay based on the response of the Nb 2 Node lymphoma cells to lactogenic hormones has been developed. Human serum stimulated cell replication. The effect was completely abolished if excess antibodies to both hPRL and hGH were present. The stimulation obtained with a number of human serum samples correlated very well with the sum of the hPRL and hGH concentrations in the sera, as determined by RIA (r = 0.95; P < 0.001). The concentrations of either hPRL or hGH in human serum could be individually determined by specifically blocking the growth stimulatory effect of the other hormone by adding excess anti-hGH or anti-hPRl. The sensitivity of this bioassay for PRL and hGH in serum exceeds that of RIAs.

Biological Assay↗
PubMed1980

Prolactin-stimulated growth of cell cultures established from malignant Nb rat lymphomas.

A malignant Nb rat lymphoma which in vivo is stimulated by estrogens has been established in suspension culture. The cultured cells grew readily in Fischer's medium supplemented with fetal calf serum (10%) and 2-mercaptoethanol (10(-4) M). If horse serum was substituted for fetal calf serum, population growth ceased; i.e., cultures became "stationary." Such stationary cultures could be induced to resume active growth by the addition of a pituitary hormone, prolactin (ovine, rat); concentrations as low as 10 pg/ml had a detectable effect. In contrast, other pituitary hormones or estrogens had little or no effect. The evidence in this and an accompanying paper suggests that prolactin (or related substances) has a role in the growth of some cancers of lymphoid origin in rats.

Animals↗
PubMed1980

Evidence in vivo and in vitro of a role for the pituitary in the growth of malignant lymphomas in Nb rats.

The growth of a transplantable malignant lymphoma in Nb rats, the Nb 2 node, was accelerated in rats bearing an estrogen pellet and, in particular, in rats bearing transplanted tumors of the anterior pituitary gland. "Lymphoma cell growth-promoting activity" in the sera from the rats was detected and assayed using cultures of Nb 2 node cells. The "activity" of the sera paralleled the degree to which lymphoma growth was accelerated in the animals. Serum from normal rats at a concentration of 1% was moderately active in stimulating culture growth; serum from estrogenized rats was at least 10 times more active; serum from rats bearing pituitary tumors was extremely active and stimulated growth even at a concentration of 0.001% (1:10(5) dilution). In contrast, serum from hypophysectomized rats was devoid of activity, even if the animals were estrogenized. The evidence indicates (a) that the growth of the lymphoma in Nb rats was stimulated by factor(s) in the peripheral blood, the levels of which were subject to control by the pituitary, and (b) that estrogen stimulated lymphoma growth indirectly, through the pituitary gland.

Animals↗