Polyamines and tumor cells: effect of transformation of chick embryo fibroblasts by Rous sarcoma virus on polyamine levels

The effect of transformation of chick embryo fibroblasts, by Rous sarcoma virus, on intracellular polyamine levels has been studied. A good correlation between spermidine and cellular protein content has been demonstrated. Upon changing the medium, a sharp increase in spermidine level was noticed both in normal and transformed cells. This increase was accompanied by enhanced protein synthesis. The intracellular concentrations of spermine and spermidine were very similar in normal and transformed cells. On the other hand, significant differences in putrescine levels were demonstrated: in normal cultures the intracellular concentration of putrescine reached a plateau approximately 6 days after seeding, whereas a continuous rise of the diamine in transformed cells was noticed. These differences, which were observed in cultured cells, may explain the known accumulation of polyamines during neoplastic growth.     

Vanadium-induced Cl(-)-secretion in rabbit descending colon is mediated by prostaglandins.

Vanadium in the 4+ (vanadyl-ion) and 5+ (vanadate-ion) oxidation state stimulates furosemide-sensitive electrogenic Cl- secretion in isolated epithelia of rabbit descending colon. This effect is associated with an increased release of prostaglandin E2 from the tissue. Inhibitors of phospholipase A2 or cyclooxygenase abolish both vanadium-induced release of prostaglandin E2 and Cl- secretion. Neuronal mechanisms are not likely to be involved, as tetrodotoxin does not affect the vanadate induced Cl- secretion. Although vanadate is known to inhibit Na+,K(+)-ATPase activity, no inhibition of active Na+ transport was observed in intact colonic epithelia suggesting a rapid intracellular reduction of vanadate ions to vanadyl ions which have no inhibitory effect on the Na+,K(+)-ATPase. The present findings therefore indicate that vanadate stimulated colonic Cl- secretion involves intracellular conversion of vanadate to vanadyl and release of prostaglandin E2.     

6;7 chromosomal translocation in spontaneously arising rat immunocytomas: evidence for c-myc breakpoint clustering and correlation between isotypic expression and the c-myc target.

Our previous studies have shown that spontaneously arising immunocytomas in the LOU/Ws1 strain of rats contain a t(6;7) chromosomal translocation in all seven tumors studied (F. M. Babonits, J. Spira, G. Klein, and H. Bazin, Int. J. Cancer 29:431-437, 1982). We have also shown that the c-myc is located on chromosome 7 (J. Sümegi, J. Spira, H. Bazin, J. Szpirer, G. Levan, and G. Klein, Nature (London) 306:497-499, 1983) and the immunoglobulin H cluster on chromosome 6 (W.S. Pear, G. Wahlström, J. Szpirer, G. Levan, G. Klein, and J. Sümegi, Immunogenetics 23:393-395, 1986). We now report a detailed cytogenetic and molecular analysis of nine additional rat immunocytomas. The t(6;7) chromosomal translocation is found in all tumors. Mapping of the c-myc breakpoints showed that in 10 of 14 tumors, the c-myc breakpoints are clustered in a 1.5-kilobase region upstream of exon 1. In contrast with sporadic Burkitt''s lymphoma and mouse plasmacytoma, only 1 of 14 tumors contains the c-myc breakpoints in either exon 1 or intron 1. Analysis of the sequences juxtaposed to the c-myc show that immunoglobulin H switch regions are the targets in at least five tumors and that there is a strong correlation between the secreted immunoglobulin and the c-myc target. Unlike sporadic Burkitt''s lymphoma and mouse plasmacytoma, at least two rat immunocytomas show recombination of the c-myc with sequences distinct from immunoglobulin switch regions.     

Photoaffinity labeling of the calcium channel antagonist receptor in the heart of the cardiomyopathic hamster

The high affinity 1,4-dihydropyridine receptors of the cardiac membrane calcium channel from Syrian Cardiomyopathic hamsters were studied using [3H] PN200-110 and [3H]azidopine as ligands. [3H]Azidopine was photoincorporated covalently into bands of 180, 100, 79, 45 and 31 kDa, as determined by SDS/polyacrylamide gel electrophoresis. Photolabeling of the 180 kDa band is protected by 2 microM [1H]PN200-110 whereas the lower Mr bands are not. Thus, only the 180 kDa band is the calcium channel linked 1,4 dihydropyridine receptor. The photoincorporation into this 180 kDa band is doubled with samples of myopathic hamsters vs. control hamsters. It is suggested that the increase in calcium channel receptors may be involved in the pathogenesis of this cardiomyopathy.     

Determination of radioligand specific activity using competition binding assays.

Radioligand binding assays are routinely utilized in laboratories throughout the world to study receptors and their related binding sites, carrier proteins, and enzymes. To accurately estimate equilibrium binding parameters, such as the equilibrium dissociation constant (Kd) and maximal number of binding sites (Bmax), the investigator must know the correct value of the specific activity of the radioligand. If the specific activity is overestimated the Kd and Bmax values will be underestimated, while underestimation of the specific activity results in an overestimation of the Kd and Bmax. The present communication describes a simple and rapid method for determining the specific activity of a radioligand using homologous competition binding assays. Performing the competition assays at two or more different concentrations of the radioligand allows the specific activity to be determined from the IC50 values without the need of analytical methods to quantify minute amounts of the radioligand. In addition to providing the specific activity, use of this method estimates the Kd for the radioligand. This method was utilized to determine the specific activity and Kd for two blockers of the dopamine uptake carrier, [3H]GBR-12935 and [3H]-CFT, which share a common binding site in the striatum.     

Declining populations of the fingernail clamMusculium transversum in the upper Mississippi River

We examined recent temporal trends in the abundance of fingernial clamsMusculium transversum (formerlySphaerium transversum) in the upper Mississippi River. Historical data on densities of fingernail clams were obtained from regional scientists and published literature. We also sampled benthos in six navigation pools in summer 1991, finding very few fingernail clams. The combined data set, including historical data and sampling results, extended from 1973 to 1992 and was sufficient to statistically evaluate trends in densities of fingernail clams in eight pools. Populations of fingernail clams declined significantly in five of the eight pools examined (Pools 2, 5, 7, 9, and 19), which spanned a 700-km reach of river from St. Paul, Minnesota, to Keokuk, Iowa. Densities in Pool 19, which had the longest historical record on fingernail clam abundance, averaged 30 000 m⁻² in 1985 and progressively declined to zero in 1990. Combined data from all eight pools showed a significant decline in abundance of fingernail clams. An evaluation of potential causal factors led us to hypothesize that the population declines in Pools 2 to 9 were linked to point-source pollution rather than to dredging activity or commercial navigation traffic. In Pool 19, the declines of fingernail clams may have resulted from low-flow conditions during drought periods, but the causal mechanisms by which low flow influences fingernail clam abundance are unclear. The decrease in fingernail clam populations may adversely affect certain fish and wildlife, such as migrating lesser scaupAythya affinis, which feed heavily on the small mollusk. Moreover, the decreases in populations of this pollution-sensitive mollusk may signal a large-scale deterioration in the health of this riverine ecosystem. The U.S. Government right to retain a non-exclusive royalty-free license in and to any copyright is acknowledged.     

Blood groups of crab-eating macaques (Macaca fascicularis) demonstrated by isoimmune rhesus monkey (Macaca mulatta) sera.

Twenty-one isoimmune sera produced in rhesus monkey (Macaca mulatta) containing type-specific antibodies for simian-type red cell antigens were tested for their cross-reactivity with red cells from crab-eating macaques (M. fascicularis). The majority of the antisera gave cross-reactions determining polymorphisms in the red cells of crab-eating macaques, homologous to those of rhesus monkeys. These results attest to the close taxonomic realationship between the two species of macaques, and have the practical implication that isoimmune sera produced for blood typing can also be used for typing red cells from related species, as has been also observed in studies on apes.     

Genetic polymorphisms in carnitine palmitoyltransferase 1A gene are associated with variation in body composition and fasting lipid traits in Yup'ik Eskimos

Variants of carnitine palmitoyltransferase 1A (CPT1A), a key hepatic lipid oxidation enzyme, may influence how fatty acid oxidation contributes to obesity and metabolic outcomes. CPT1A is regulated by diet, suggesting interactions between gene variants and diet may influence outcomes. The objective of this study was to test the association of CPT1A variants with body composition and lipids, mediated by consumption of polyunsaturated fatty acids (PUFA). Obesity phenotypes and fasting lipids were measured in a cross-sectional sample of Yup'ik Eskimo individuals (n = 1141) from the Center of Alaska Native Health Research (CANHR) study. Twenty-eight tagging CPT1A SNPs were evaluated with outcomes of interest in regression models accounting for family structure. Several CPT1A polymorphisms were associated with HDL-cholesterol and obesity phenotypes. The P479L (rs80356779) variant was associated with all obesity-related traits and fasting HDL-cholesterol. Interestingly, the association of P479L with HDL-cholesterol was still significant after correcting for body mass index (BMI), percentage body fat (PBF), or waist circumference (WC). Our findings are consistent with the hypothesis that the L479 allele of the CPT1A P479L variant confers a selective advantage that is both cardioprotective (through increased HDL-cholesterol) and associated with reduced adiposity.