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When incubated with cultured mouse neuroblastoma cells under growth stimulatory condition, [3H]putrescine or [3H]spermidine can metabolically label a cellular protein of apparent molecular mass 18 kDa. The labeling, which leads to hypusine formation, is due to a covalent linkage between a lysine residue and the butylamino group derived from spermidine. This reaction can be demonstrated in the cytosolic fractions obtained from cells whose spermidine pool was depleted by prior treatment with alpha-difluoromethylornithine. In an effort to characterize the enzyme system involved in this unique post-translational modification, we found that NAD+ at 0.1 mM stimulated labeling more than 150-fold. Other nucleotides such as NADP+, ATP and GTP were ineffective. The fact that NAD+ dramatically stimulated labeling of the 18 kDa protein indicated that the enzyme involved in hypusine formation may be an NAD+-requiring enzyme.

Neurite outgrowth from chick embryonic sensory ganglia explants was induced by activators of protein kinase C and other compounds known to stimulate the hydrolysis of inositol phospholipids. The addition of diacylglycerols, phospholipase C and muscarine chloride to a defined growth medium promoted the outgrowth of dense neurites from ganglia explants which were morphologically distinct from those induced by the phorbol ester TPA. Moreover, these neurite-promoting agents did not enhance non-neuronal cell proliferation and were ineffective in the absence of insulin and/or progesterone.

The effect of polyamines (putrescine, spermidine and spermine) on endogenous protein phosphorylation in mouse neuroblastoma cells was investigated by using techniques of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. The results indicated that spermine at 1mM completely inhibited the phosphorylation of the 11,000-dalton and 120,000-dalton proteins in nuclear fractions. The inhibition of the phosphorylation of the 11,000-dalton but not the 120,000-dalton protein by spermine was also observed in five other cell lines examined and appeared to be a general phenomenon. The inhibitory effect of spermine on the phosphorylation of the 11,000-dalton protein was specific, other cations such as ammonium chloride, arginine, putrescine, cyclen and trien were ineffective at equal molar or much higher concentrations.

The acute effects of Nifedipine on hepatic venous pressure gradient (HVPG) and estimated hepatic blood flow (EHBF) were investigated in six patients with HBsAg-positive cirrhosis and portal hypertension by simultaneous hepatic venous catheterization. The mean arterial pressure significantly decreased by 18% at 1 h after sublingual administration of 10 mg Nifedipine. Significant increases in heart rate (13%) and cardiac output (20%) were also demonstrated in our patients 1 h after administration of Nifedipine. However, there were no significant changes in HVPG and EHBF after Nifedipine. We conclude that in patients with HBsAg-positive cirrhosis who receive Nifedipine, EHBF is maintained despite a substantial change in systemic hemodynamics, and that Nifedipine is not effective in acutely reducing HVPG.

This is a study of the regulation of expression of the RI cAMP-binding protein in mouse neuroblastoma cells as it relates to neurotransmitter phenotype and neurite outgrowth. Dibutyryl cAMP was used to promote differentiation of the cholinergic NS-20, the adrenergic N1E-115, the neurotransmitter-inactive N-18, and the neurite-minus N1A-103 mouse neuroblastoma cells. The amount of the RI cAMP-binding protein was quantitated by photoaffinity labeling of the 47,000-dalton RI protein with 8-N3-[32P]cAMP and by Western blot, ELISA, and immunocytochemistry. Our results showed that dibutyryl cAMP induced the RI cAMP-binding protein by three to fivefold in each of the four neuroblastoma cell lines examined. The increased expression of the RI cAMP-binding protein was not linked to neurite outgrowth, a parameter of morphological differentiation in the neuroblastoma cells. Thus, the RI cAMP-binding protein can be induced in the neurite-minus N1A-103 neuroblastoma round cells; further, 8-bromo-cAMP effected neurite outgrowth without inducing the RI cAMP-binding protein in the neurite-positive cell lines. Indirect immunocytochemistry of RI showed a cytoplasmic localization with little evidence of nuclear staining. The increase in RI cAMP-binding protein coincided with an increase in the cAMP-phosphodiesterase and a decrease in cAMP-dependent phosphotransferase activity in the mouse neuroblastoma cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Experimental studies of I-131-LYM-1 MoAb radioimmunotherapy (RIT) and LYM-1 MoAb immunotherapy (IT) were performed in 23 implanted nude mice model. 1 x 10(7) human B-cell lymphoma cells (Raji Burkitt's cells) were injected subcutaneously into the thigh. Three weeks later, they were divided into four groups: Group A (N = 6), as control group; Group B (N = 5), received LYM-1 MoAb 60 micrograms, as IT group; Group C (N = 7), received I-131-LYM-1 MoAb 120 mu ci; and Group D (N = 5), received I-131-LYM-1 MoAb 240 mu ci. Radioimaging was performed on day 7,8,9 postinjection. Tumor growth after 30 days was as follows: Group A, from 0.040 +/- 0.091 CM3 to 7.638 +/- 5.925; Group B, from 0.326 +/- 0.315 to 4.498 +/- 2.572; Group C, from 0.632 +/- 0.449 to 3.330 +/- 4.875; Group D, from 0.490 +/- 0.531 to 2.352 +/- 2.188. When converted to Relative Increase of Tumor Volume (Times-X): The tumor in Group A increased 4346.19 X; in Group B 312.68 X; in Group C 15.94 X; and in Group D 8.36 X. Significant differences were noted between Groups A and B (p less than 0.05), A and C (p less than 0.025), A and D (p less than 0.025), B and C (p less than 0.025) and B and D (p less than 0.025). The difference between Group C and D was insignificant (p greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

A series of chemical extraction procedures are used to obtain data on the partitioning of trace metals among the various geochemical phases of sediments. These components include intersititial water, solubility of solid minerals, ions on exchange sites, metal carbonates, easily reducible phases, organics and sulfides, iron oxides, and lithogenous (mineral residual) fractions. In general, a mass balance of less than 10% deviation can be obtained. Experimental results show very small fractions of trace metals to be in the form of interstital water or soluble ions. Trace metals in the exchangeable phase are almost negligible, and those in the mineral residual phases range from 2.5% Cd for one sediment to 98% Cu for another. The non-residual trace metals content is found to increase with decreasing sand content.

Possible changes of glycoproteins in IMR-90 human embryonic lung fibroblasts during senescence in vitro were studied by the metabolic labeling technique using radioactive precursors for carbohydrate moieties of glycoproteins. IMR-90 fibroblasts at three different population doubling level (PDL) were incubated with [3H]fucose and [3H]glucosamine for various periods of time. The radioactively labeled glycoproteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography. The results indicated a marked increase, by more than eight-fold on per mg protein basis, of labeling by [3H]fucose in old IMR-90 fibroblasts (PDL = 45) as compared to young (PDL = 22) and middle-age (PDL = 30) IMR-90 fibroblasts. In contrast, no significant difference in [3H]glucosamine labeling was observed in young and old IMR-90 cells.

A method is described that permits the rapid extraction of the cell surface glycoproteins of two murine leukemic cells, the P-388 and the L-1210 cells as well as those of the human adenocarcinoma cells, the HeLa cells. Proof of the surface location of these glycoproteins is provided by labeling the intact cells; (a) with 125I by the lactoperoxidase iodination technique; (b) with 3H by the galactose oxidase-reductive tritiation method. Most of these glycoproteins were also shown to incorporate radioactive glucosamine and fucose. By these criteria as well as by the distribution of molecular weights, the surface glycoproteins of the two murine cells are indistinguishable; however, they differ markedly from the surface glycoproteins of HeLa cells. The extracts of the murine cells were shown to contain lectin receptor activity as determined by their ability to inhibit the lectin-induced agglutination of the intact cells.

The minimum requirements for eliciting or enhancing ornithine decarboxylase activity (EC. 4.1.1.17); L-ornithine carboxylase) in neuroblastoma cells incubated in salts-glucose solutions have been investigated. These incubation conditions permit the study of changes in ornithine decarboxylase activity independently of the growth-associated reactions that occur in cell culture media (Chen, K.Y. and Canellakis, E.S. (1977) Proc. Natl, Acad. Sci. U.S.A. 74, 3791-3795). Ornithine decarboxylase activity can be elicited by a variety of asparagine and other amino acid analogs, including alpha-aminoisobutyric acid, that cannot participate in protein synthesis. Of the eleven asparagine analogs tested, alpha-N-CH3-DL-asparagine is the most potent in eliciting ornithine decarboxylase activity and is equivalent to asparagine in this regard. Inclusion of polar groups into the asparagine molecule results in the loss of its ability to elicit ornithine decarboxylase activity. With the use of these analogs and of analogs of other amino acids it is shown that the rapid fall in ornithine decarboxylase activity that is noted following cycloheximide treatment may not be a consequence of the inhibition of protein synthesis. The rapid fall in ornithine decarboxylase activity is primarily due to the removal of the agent that elicits and stabilizes its activity. These results, the finding that alpha-aminoisobutyric acid stimulates ornithine decarboxylase activity and that sodium is required for the stimulation of ornithine decarboxylase activity are discussed in relation to the "A" amino acid transport system.

Aging of IMR-90 human diploid fibroblasts in vitro is accompanied by significant changes of polyamine metabolism, most notably, a 5-fold decrease of serum-induced activity of ornithine decarboxylase, the key enzyme in the biosynthesis of polyamines (Chen, K. Y., Chang, Z. F., and Liu, A. Y.-C. (1986) J. Cell. Physiol. 129, 142-146). In this paper, we employed Northern blot hybridization and affinity radiolabeling techniques to investigate the molecular basis of this age-associated change of ornithine decarboxylase activity. Since the induction of ornithine decarboxylase by serum is a mid-G1 event, we also examined expressions of other cell cycle-dependent genes that are induced before and after the mid-G1 phase to determine if their expressions may also be age-dependent. Our results demonstrated a 3-fold decrease of the amount of active ornithine decarboxylase molecules that can be labeled by alpha-difluoromethyl[3H]ornithine in senescent IMR-90 cells (population doubling level (PDL) = 52) as compared to young cells (PDL = 22). However, the levels and kinetics of induction of ornithine decarboxylase mRNA in both young and senescent IMR-90 cells were found to be identical throughout a 24-h time period after serum stimulation. The time course and the magnitude of the expression of c-myc, an early G1 gene, were quite similar in young and senescent IMR-90 cells and appeared to be PDL-independent. In contrast, the expression of thymidine kinase, a late G1/S gene, was significantly reduced in senescent IMR-90 cells. Levels of thymidine kinase mRNA and thymidine kinase activity in senescent IMR-90 cells were 6- and 8-fold less than those in young cells, respectively. Based on these data, we proposed that impairment of cell cycling in senescent IMR-90 cells may occur at the late G1/S phase and that decreases of ornithine decarboxylase activity and putrescine accumulation during cell senescence may contribute to this impairment.

Mouse leukemia L-1210 cells were iodinated with 125I; this permitted the development of a method for the isolation of the plasma membranes. These show a 10- to 16-fold increase in the specific activity of 125I over that of the cell homogenate and a 20-fold increase in the specific activities of 5'-nucleotidase and alkaline phosphatase; 20-fold increase in the specific activities of 5'-nucleotidase and alkaline phosphatase; no mitochondrial or microsomal marker enzyme activities were detected. Sodium dodecyl sulfate gel electrophoresis of the plasma membranes shows approx. 40 peptides with molecular weights ranging from 10 000 to over 200 000; a polypeptide (Mr 50 000) predominates. Of 13 iodinated surface membrane proteins, the major radioactive peptide has a molecular weight of 85 000. The importance of the selection of the appropriate gel system for the analysis of membrane proteins is emphasized.