Common characteristics of normal and cancerous growth in vivo.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
O Greengard
Publications and source records attributed to O Greengard.
Enzymic composition and growth rate of human pleural mesothelioma transplants in nude mice.
The enzymic composition of 7 human mesothelioma lines propagated in nude mice was compared with 4 of the original and 15 additional mesotheliomas sampled during the patients' surgery. The xenografts exhibited several-fold higher thymidine kinase (TK), uridine kinase (UK), phosphoserine phosphatase (PSP) and peptidyl proline hydroxylase (PPH) concentrations than the fresh human samples, while their DNA, gamma-glutamyl transpeptidase (GGT) and beta-galactosidase (Bgal) contents remained similar. The volume growth rate of the xenografts (doubling time, DT = 9.23 +/- 1.25 days) was much faster than that of tumors in the human host, and the decline of this rate with increasing nodule size was accompanied by decreases in TK and PSP concentrations. This first quantitative biochemical study of xenografted human neoplasms indicates that 1) pleural mesotheliomas, though preserving their histological characteristics after heterotransplantation, show considerable increases of enzymes in nucleic acid, collagen, and nonessential amino acid synthesis, and that 2) the concentration of TK is a good indicator of the different growth properties of tumors in a mouse rather than in the human host.
Animals↗
Neutrophil maturation and hypersegmentation promoted in normal bone marrow by a carcinoma-elaborated protein factor.
In rats with subcutaneously transplanted mammary carcinoma 5A (MC) and 2 to 10-fold elevations in the blood content of mature neutrophils, 30-50% of the neutrophils showed pronounced hypersegmentation. This phenomenon could be reproduced in liquid culture of bone marrow cells from normal (tumor-free) animals by 48 hr incubation with the MC host's serum, or with MC-conditioned medium whose activity was attributable to an over 50,000 MW protein. The effects in vitro, occurring without change in total cell number and accompanied by increases in gamma-glutamyltranspeptidase (GGT) and alkaline phosphatase (AP) activity, included decreases in the percents of progenitors (myeloblasts, promyelocytes, myelocytes, metamyelocytes and bands) and an increase in mature neutrophils 50% of which exhibited obvious hypersegmentation. Much less if any neutrophil hypersegmentation, and no statistically significant decrease in immature cells, occurred in response to the several colony stimulating factors (CSFs) tested, although (in addition to inducing GGT and AP) some CSFs did cause an increase in mature neutrophils. These investigations demonstrate the efficacy of a MC-elaborated blood-borne protein to promote myeloid cell maturation, and describe a system for the first time in which neutrophil hypersegmentation can be studied in vitro.
Animals↗
Mitogen activation of human lymphocytes from normal and acute lymphocytic leukemia (ALL) subjects.
Lymphocytes from acute lymphocytic leukemia (ALL) subjects were converted by mitogens to blast-like cells whose microscopic appearance and rate of formation was indistinguishable from those in mitogen incubated control lymphocytes. In ALL lymphocytes, however, pokeweed mitogen (PWM) failed to stimulate GGT expression; the mean increase it caused in thymidine kinase (TK) activity and thymidine incorporation was normal, though there were appreciable individual variations. These variations were also apparent with concanavalin A (Con A) but, in most ALL cases, TK and thymidine incorporation rose to much higher levels than in Con-A-treated control lymphocytes. The results indicate that evaluation of the response to mitogens by quantitative biochemical criteria provides a sensitive method for revealing functional impairments in microscopically normal ALL lymphocytes.
Cells, Cultured↗
gamma-Glutamyltranspeptidase activity in intact leukocytes: flow cytometric analysis and sorting.
A fluorescent method developed for visualizing gamma-glutamyltranspeptidase (GGT) in intact liver cells was adapted to leukocytes and used in a multiparameter flow cytometric study of blood and bone marrow cells from rats with subcutaneous implants of mammary carcinoma 5A. The severe granulocytosis caused by this non-metastatic tumor was preceded by a progressive rise in the percentage of leukocytes with high GGT fluorescence. Both granulocytes and small, immature cells of bone marrow showed increased GGT expression, whereas in blood this increase was attributable entirely to mature granulocytes. At 28 days (but not yet at 14 days) after carcinoma implantation, 20-30% of blood or bone marrow granulocytes constituted a distinct subpopulation in that their GGT fluorescence intensity range was much higher and did not overlap with the range for the rest of the population. The results indicate that fluorescent GGT assay of intact leukocytes provides a useful probe for flow cytometric analysis of population heterogeneity in leukoproliferative disorders.
Animals↗
Phenylalanine uptake in neonatal and infant rat brain.
At the same severe elevations in blood phenylalanine (Phe) levels maintained for 4 h, much higher cerebral Phe concentrations were found in 4-day-old than in 16- or 70-day-old rats. In order to compare this developmental change with 14C-Phe influx mediated by the L transport system, the rapid intracarotid injection method was adapted for use in neonatal rats. The brain uptake index (BUI) thus determined for the first time through the suckling period was significantly higher on the 4th day of age than on the 7th or 24th day, while no significant change occurred during subsequent life. This early period of change in influx across the blood-brain barrier overlapped with the age period of decrease of the hyperphenylalaninemia-associated accumulation of Phe in the brain. The results indicate that by the time when intermittent feeding begins, the brain has developed a considerable ability (a) to protect itself against physiological (e.g. postprandial) fluctuations in circulating Phe levels, and (b) to restrict the cerebral accumulation of Phe from pathologically elevated blood concentrations such as those in phenylketonuria.
Animals↗
Enhancement of gamma-glutamyl transpeptidase expression in bone marrow cells by colony stimulating factors.
Studies on blood serum from mammary carcinoma (MC) hosts, which promoted gamma-glutamyl transpeptidase (GGT) expression by normal rat bone marrow cells in liquid culture, were extended to various granulocyte-macrophage colony stimulating factors (CSFs). GGT concentration per cell was found to increase (without change in total cell number) by incubation for 48 h with purified CSF-2 gamma and CSF-1 (but not interleukin-3), with human giant cell elaborated GM-CSF and L-cell conditioned medium, as well as with the 3 MC preparations (host serum, MC extract and MC conditioned medium). GGT-inducing ability (per milligram protein) ranked the 7 preparations in the same order as did their proliferative effect (number of colonies per milligram protein) in the standard mouse bone marrow agar culture system. The quantitative correlation between these two kinds of activities (linear for their logarithmic values) was highly significant, r = 0.976, p less than 0.001. The alkaline phosphatase concentration of bone marrow cells in liquid culture was also increased in the presence of the same 7 preparations, and this again was proportional (r = 0.985, p less than 0.001) to their colony stimulating potential.
Animals↗
Characterization of a mammary carcinoma elaborated factor stimulating gamma-glutamyltranspeptidase expression in bone marrow cells.
The elevation of bone marrow gamma-glutamyltranspeptidase (GGT) and alkaline phosphatase (AP) content in rats carrying mammary carcinoma 5A (MC), reproduced in a short-term (48-h) liquid culture of normal bone marrow cells, was found to be attributable to a blood-borne protein factor with an apparent molecular weight of 60,000. Partial purification, based on the extent of stimulation of GGT expression in this culture, increased the specific activity of the host serum from 1.5 to 40 units and that of MC extracts from 6 to 560 units. Production of the factor by MC in vitro, however, resulted in specific activities of 3000 units in the conditioned medium, and a further 60-fold purification was achieved by DEAE-cellulose, Sephadex G-100, and hydroxylapatite chromatography. The chemical characteristics of the MC-elaborated protein indicate nonidentity to previously isolated colony formation stimulating factors which also induced GGT (and AP) expression by rat bone marrow cells. Most of the AP inducing ability of the MC-serum and MC-conditioned medium copurified with and was still present in preparations with the highest specific activity vis à vis GGT. In mouse (instead of rat) bone marrow cells, however, no AP response accompanied the stimulation of GGT expression by MC (or colony formation stimulating factor) preparations.
Alkaline Phosphatase↗
Cerebral serotonin regulation by phenylalanine analogues and during hyperphenylalaninemia.
Severe hyperphenylalaninemia induced in infant rats by 3 days of treatment with p-chlorophenylalanine (p-cl phe) plus phenylalanine (phe) did not lower the tryptophan concentration of the brain, and the cerebral serotonin (5-HT) deficiency was attributable entirely to the known suppression to tryptophan hydroxylase (TPH) by p-cl phe. The decrease in 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) was thus no more pronounced than in rats which, treated with p-cl phe alone, were devoid of hyperphenylalaninemia. Suppression of TPH was found to also underlie the decrease in cerebral 5-HT caused by treatment with alpha-methylphenylalanine (alpha-mephe) alone: a 22% loss of midbrain TPH activity was detectable 24 hr after an injection only, reverted toward the normal during the next 2 days, and was clearly unrelated to the weak competitive inhibition of the enzyme by alpha-mephe in vitro. However, alpha-mephe (unlike p-cl phe), when administered together with phe, did not suppress TPH, nor did it counterbalance the reduction of cerebral tryptophan uptake by excess phe. Thus the 5-HT diminution in the rat model of phenylketonuria produced by treatment with alpha-mephe plus phe was attributable to hyperphenylalaninemia and the inhibition of tryptophan transport to the brain. Injection of tryptophan was found to restore the cerebral 5-HT level in the face of persistently severe hyperphenylalaninemia.
Animals↗
Diminished lymphocyte and granulocyte gamma-glutamyltranspeptidase activity in acute lymphocytic leukemia and response to chemotherapy.
gamma-Glutamyltranspeptidase (GGT) activity (per mg protein) in blood lymphoid cells of 27 children with acute lymphoblastic leukemia (ALL) (1.05 +/- 0.15) was significantly below that of controls (2.25 +/- 0.30), became normalized during chemotherapy-induced remission (2.47 +/- 0.26), and was low again (1.59 +/- 0.62) in relapsed subjects. Individual variations in the GGT activity of the blood lymphoid cell fraction (per mg protein) bore a significant inverse correlation to the number of white blood cells (WBC) as well as of blasts per ml blood. Blasts had minimal GGT activity; however, partial GGT deficiency was also exhibited by the microscopically normal circulatory lymphocytes of several patients prior to treatment and in relapsed subjects whose blood was still devoid of blasts. Significantly diminished GGT activity (per mg protein) was found in the blood granulocytes of ALL subjects. This deficit, restored during remission and present again at relapse, varied in magnitude but showed no statistically significant correlation to the different patients' degree of neutropenia. In about one-third of the newly diagnosed or relapsed pre-B ALL children, the circulatory granulocytes' GGT activity was only 10-20% of normal. The results suggest that 1) the presence or absence of this sign of functional maldifferentiation in granulocytes is a factor in the heterogeneity of disease manifestation among subjects with apparently the same type of ALL and that 2) measurement of GGT in the circulatory granulocytes, as well lymphocytes, may be useful for monitoring the efficacy of chemotherapy.
Bone Marrow↗
Cerebral glycine content and phosphoserine phosphatase activity in hyperaminoacidemias.
Chronic hyperphenylalaninemia maintained with the aid of a suppressor of phenylalanine hydroxylase, alpha-methylphenylalanine, increases the glycine concentration and the phosphoserine phosphatase activity of the developing rat brain but not that of liver or kidney. Similar increases occur after daily injections with large doses of phenylalanine alone, while tyrosine, isoleucine, alanine, proline, and threonine, were without effect. Treatment with methionine, which increases the phosphoserine phosphatase activity of the brain and lowered that of liver and kidney, left the cerebral glycine level unchanged. When varying the degrees of gestational or early postnatal hyperphenylalaninemia, a significant linear correlation was found between the developing brains' phosphoserine phosphatase and glycine concentration. Observations on the uptake of injected glycine and its decline further indicate that coordinated rises in the brain's phosphoserine phosphatase and glycine content associated with experimental hyperphenylalaninemia denote a direct impact of phenylalanine on the intracellular pathway of glycine synthesis in immature animals.
Age Factors↗
Lymphocyte thymidine kinase and treatment response in acute lymphocytic leukemia.
The activity of thymidine kinase (TK) and the proportion of its isozymes (TK1/TK2) were studied in peripheral lymphoid cells of 37 children with acute lymphocytic leukemia (ALL). The high TK in 25 untreated subjects (31.5 +/- 8.9) decreased during chemotherapy-induced remission to uniformly low (5.3 +/- 0.4) normal values, and rose again during relapse to a mean of (24.8 +/- 8.1). The proportion of isozyme 1 followed the same pattern but TK was a more sensitive indicator of disease state. The lymphocyte fractions' TK (per mg protein) correlated with the number (per ml blood) of WBCs, blasts and lymphocytes. Although the higher TK of blasts than of apparently normal lymphocytes was confirmed in cases permitting clean physical separation, the lymphocyte fraction of several untreated subjects with minimal blast counts also exhibited elevated TK. Moreover, this elevation was also seen in relapsed cases even if their blood (unlike bone marrow) was devoid of blasts. The results indicate that quantification of TK can reveal a subpopulation of maldifferentiated lymphocytes which are microscopically normal and that it may provide an objective parameter of prognostic differences between ALL subjects with similar hematological characteristics.
Adenosine Triphosphate↗
Enzyme pathology of human mesotheliomas.
In samples of 16 surgically resected mesotheliomas arising from the pleura of the human lung, 6 enzymes from different metabolic pathways, DNA, and mitotic frequency were quantified. The mesotheliomas, irrespective of cell type or grade, showed lower gamma-glutamyl transpeptidase (GGT) concentration than 36 of the 38 pulmonary adenocarcinomas. The mean concentration of this enzyme in the 15 mesotheliomas was an eighth of that in the 56 carcinomas, whereas their DNA content was similar. The quantitative correlation of thymidine kinase (TK), uridine kinase (UK), and phosphoserine phosphatase to mitotic frequency was highly significant for mesotheliomas, as well as for carcinomas. As estimated from their TK [and its recently established quantitative correlation to volume doubling time (DT)], the DT of the 16 mesotheliomas ranged from 50 to over 700 days, with a somewhat longer median than the median for pulmonary carcinomas. Subject survival, though shortest for the 2 sarcomatous mesothelioma cases, varied over an overlapping range for mesotheliomas with epithelial or mixed cell type. The biopsy samples' TK and UK concentrations, however, showed a significant inverse correlation with months of survival after diagnosis. Survival time after the first appearance of symptoms decreased linearly (on log scales) with TK concentration (P less than .001) over the 14 cases. The results of this first quantitative study of a spectrum of biochemical constituents of mesotheliomas identify GGT as an enzyme whose measurement guards against mistaking mesotheliomas and adenocarcinomas for one another and show that the TK concentrations of these mesothelioma samples bear a highly significant, inverse correlation to the postdiagnosis survival time of the individual subjects.
Biopsy↗
Pulmonary carcinoid tumors: enzymic discriminants, growth rate, and early age of inception.
Analyses of enzymes from various metabolic pathways in pulmonary carcinoid tumors and radiological measurements of their volume increase were compared with those for lung carcinomas of various cell types. The results describe new biochemical features in carcinoid tumors, present the first quantitative evidence for their slow growth rate (i.e., long doubling time) in vivo, and show that measurement of 2 or 3 appropriate enzymes in biopsy samples can guard against instances in which carcinoids and adeno- or oat cell carcinomas are mistaken for one another on histological examination. The uridine kinase to thymidine kinase ratio as well as the beta-galactosidase concentration of carcinoid tumors were 5 times higher than of carcinomas, and their gamma-glutamyl transpeptidase was below that of all 35 adeno- and the 11 squamous cell carcinomas. Thymidine kinase, which bears a quantitative inverse correlation to volume doubling time (irrespective of cell type), had much lower titers in the 9 carcinoids than in the 6 oat cell carcinomas and reflects most clearly their very different (despite common histogenesis) clinical malignancy. Owing to their long doubling time, carcinoid tumors on the average required a much longer period (40.5 years) to attain final volume than did carcinomas (17.8 years). The calculated mean age of the subjects when growth began, -0.5 years (as opposed to 51 years for carcinomas), suggests a prenatal or early childhood inception for pulmonary carcinoid tumors.
Carcinoid Tumor↗
Mechanical responses of developing Fisher rat heart. Effects of steroid hormone.
Age-dependent changes in the mechanical responses of developing Fisher rat heart during the first three postnatal weeks were studied in relation to the hypothesis that the abnormality observed in the mechanical responses of the rat heart might be calcium related. Therefore the effect of frequency of stimulation as well as the response to calcium, epinephrine and ouabain on hearts of untreated and cortisol-treated rats was compared. The positive force-frequency response observed in fetal rat heart reverted to a highly negative response by the 12th to 14th postnatal day. The biphasic mechanical responses directly paralleled reported changes in circulating glucocorticoid levels in developing rat. The force-frequency response was maximally negative when the circulating levels of glucocorticoids were lowest. The reversion of the negative force-frequency responses coincided with a gradual increase reported in the circulating levels of glucocorticoids. The negative force-frequency response was absent in the cortisol-treated developing rat heart and a definite positive pattern was observed as the rats developed. A high sensitivity to free calcium concentration, seen in control fetal and and newborn hearts, diminished after the second postnatal week. By the third postnatal week, the sensitivity to high extracellular calcium concentrations was significantly reduced. The sensitivity to calcium persisted in the cortisol-treated hearts during the 3 postnatal weeks. Cortisol-treated hearts were more responsive to epinephrine than controls. The abbreviation of time to peak tension, a hallmark of the catecholamine effect, was observed at a younger age in the cortisol-treated hearts. Cortisol-treated hearts were more responsive to the inotropic effects of ouabain than controls. The possible involvement of glucocorticoids in the control of calcium handling elements of the myocardium is discussed.
Animals↗
Biochemical measure of the volume doubling time of human pulmonary neoplasms.
The volume doubling time (DT) of human lung neoplasms, determined from sequential, presurgery roentgenograms, was compared with biochemical and histologic observations on biopsy samples of the same tumors obtained during surgery. The DTs of the 16 neoplasms ranged from 24 days in an oat cell carcinoma to over 500 in a pulmonary carcinoid tumor, and showed a statistically significant, inverse correlation to the TK (thymidine kinase) and the UK (uridine kinase) concentration of the biopsy samples per g wet weight or mg DNA. Log DT bore a linear relationship to log TK (r = 0.75, P = 0.0008) and to log UK (r = 0.69, P = 0.0067), and an even better fit to the straight line was found when plotting the log of the standardized average of the two enzymes against log DT. The results demonstrate the feasibility of a biochemical method for determining the unknown DT of the many tumors that are not amenable (on account of shape, location, or lack of prior chest x-rays) to the direct, radiologic determination of this useful, dynamic parameter of clinical malignancy.
Biopsy↗
Developmental changes of cerebral phenylalanine uptake from severely elevated blood levels.
Brain phenylalanine concentrations at plasma levels raised to that in phenylketonuric subjects were studied in rats from fetal through postnatal life. Suppression of the hepatic phenylalanine hydroxylase with alpha methylphenylalanine, and injections of age-adjusted doses of phenylalanine on the next day, assured the persistence of the same elevation of plasma levels for at least four hours prior to assay. The net phenylalanine uptake determined under these conditions underwent several-fold decreases between the fourth day and the end of the suckling period, and by about the age of 30 days it was as low as in adulthood. The development of transport properties studied here could contribute to the change with age in the vulnerability of the brain to the same degree of hyperphenylalaninemia and, since the cerebral phenylalanine uptake may decrease to non-damaging levels during childhood, it is pertinent to defining the age at which the rigorous diet of phenylketonurics might be safely relaxed.
Age Factors↗
Responses of bone marrow gamma-glutamyltranspeptidase and alkaline phosphatase in vitro to tumor-elaborated granulocytopoietic factors.
Evidence has been obtained for the humoral mediation of the recently noted tumor-induced rise of the host bone marrow gamma-glutamyltranspeptidase (gamma GT) and alkaline phosphatase (AP) content in vivo: normal rat bone marrow suspensions, if incubated for 18 hr to 3 days with serum from mammary carcinoma hosts, show 2- to 8-fold elevations (per cell) of the same 2 enzymes. The active substance(s) is in the acid-stable, HCI-ethanol-soluble polypeptide fraction of the mammary carcinoma extract, and of the hosts' blood serum. The larger the size of the neoplasm, and the faster its growth rate, the greater the effect of the host serum on the gamma GT and AP of the normal bone marrow cells. In host rats in vivo, this response is followed by increases in the number (as well as the gamma GT and AP content) of circulating granulocytes. Therefore, a positive response on the part of these enzymes in the bone marrow suspension was also sought, and found, upon incubation with preparations which enhance granulocyte colony formation in agar cultures (i.e., colony-stimulating factor and serum from endotoxin-treated rats). The results indicate: (a) that the increase in gamma GT and AP is a necessary prelude to stimulation of granulocyte multiplication by appropriate growth factors; and (b) that measurement of these enzymes in the short-term liquid culture offers a biochemical test for such factors elaborated by cancers or in nonneoplastic conditions.
Alkaline Phosphatase↗