A study of the toxic hazard that might be associated with the consumption of green potato tops.

Eating green potatoes has reportedly led to poisoning attributed to potato glycoalkaloids (PGA), primarily alpha-solanine and alpha-chaconine. Concentrations of PGA increase during the greening of potatoes but are reportedly much higher in potato tops (leaves). As it is known that members of the UK Bangladeshi community consume potato tops, a study of the toxic hazard that may be associated with the consumption of green potato tops has been carried out. PGA in seven potato varieties were determined by HPLC. Tubers protected from light contained 0.05-0.65 mg/100 g alpha-solanine and 0.3-0.63 mg/100 g alpha-chaconine. Concentrations in leaf samples ranged from 0.64 to 22.6 mg alpha-solanine/100 g and 0.06 to 55.7 mg alpha-chaconine/100 g. Aqueous leaf extracts were cytotoxic to Chinese hamster ovary cells and lysed human, rat and hamster blood cells with no difference in sensitivity among species. Oral administration of potato tops to rats, mice and Syrian hamsters had no adverse effects at the highest practicable dose. A mixture of alpha-solanine and alpha-chaconine (1:1, w/w) given orally at doses of up to 50 mg/kg body weight to hamsters had no effect, but a single ip injection of 25 mg/kg body weight or greater was lethal, with bleeding in the gut. High concentrations of cytotoxic PGA were found in some potato tops, but their effect in laboratory animals was minimal. It is concluded that the consumption of moderate quantities of potato tops (2-5 g/kg body weight/day) is unlikely to represent an acute health hazard to humans.

Distribution of glutamine synthetase and glial fibrillary acidic protein and correlation of glutamine synthetase with glutamate decarboxylase in different regions of the rat central nervous system.

The concentration of soluble glial fibrillary acidic (GFA) protein and the specific activity of glutamine synthetase (GS) were estimated in 11 central nervous system (CNS) regions of the 90-day-old rat. Marked differences were observed in the regional distribution of these astrocyte marker proteins. The striatum and spinal cord contained the lowest concentration (per g wet weight) of GFA protein and GS activity, respectively, while the olfactory bulbs had the highest level of both astrocytic proteins. Differences between the lowest and the highest values were 3-fold for GS and 4-fold for GFA protein. More significant was the marked variation in the ratio of GS to GFA protein in different CNS regions; the highest and lowest values were in the striatum and the spinal cord respectively, and the difference between the highest and the lowest value was about 5-fold. The spinal cord contained low GS and high GFA protein; on the other hand, the colliculi had high GS and relatively low GFA protein. Immunochemical detection of GS and GFA proteins in whole homogenates of different regions showed that the variation of the specific activities of GS and the concentration of soluble GFA protein were due to the differences in their absolute protein concentrations. In different regions of the brain the activity of GS was significantly correlated with that of glutamate decarboxylase, but not with that of choline acetyltransferase. These observations provide further evidence for differing biochemical properties of astrocytes from various CNS regions and for the involvement of GS in processes associated with amino acid neurotransmission.

Regulation of in vivo glutamine synthetase activity by glucocorticoids in the developing rat brain.

Induction of glutamine synthetase in vivo by glucocorticoids was studied in different brain regions of the rat during development. Corticosterone treatment resulted in an age-dependent increase in glutamine synthetase activity. In 11-day-old rats, in comparison with controls, the increase was about 80% in the cerebellum, 50% in the olfactory bulbs and 20% in the forebrain. During development the effect diminished markedly and at day 20 a marked increase was detectable only in the cerebellum, suggesting that the elevation of glutamine synthetase was dependent on the maturational state of the region at the time of hormone treatment. The increase in enzyme activity was dose-dependent, and was also observed after treatment with dexamethasone but not with testosterone, estradiol or progesterone. Pre-treatment with the protein synthesis inhibitor anisomycin blocked the hormonal response. Also, immunochemical detection of glutamine synthetase in brain homogenates fractionated by SDS-polyacrylamide gel electrophoresis and transferred to diazobenzyloxymethyl paper showed that the increase in enzyme activity was due to induction of protein synthesis. This may be the first report showing a regulation by glucocorticoids of mammalian brain glutamine synthetase in vivo, and it is in good agreement with previous findings with chick retina in vivo and various avian and mammalian cells in vitro.

Immunochemical detection of chromosomal protein HMG-17 in chromatin subunits.

Chromosomal protein HMG-17, purified from calf thymus, has been used to elicit specific antibodies in rabbits. Specific serological reaction between the antigen and the antisera is demonstrated by solid-phase radioimmunoassay and by competitive inhibition assays. The antisera did not cross-react with histones or other chromosomal HMG proteins. The antisera bound specifically to chromatin subunits isolated from HeLa cells, demonstrating that it may be used to study the in situ organization of this chromosomal protein. Chromatin purified from HeLa nuclei was digested with micrococcal nuclease, and the resulting mono- and oligonucleosomes were fractionated on a sucrose gradient. Analyses of the content of chromosomal proteins HMG-1, HMG-17, and H4 in different size nucleosomal particles, by the solid-phase radioimmunoassay, reveal that the distribution of HMG-17 was the same as that of H4, but different from that of HMG-1.

Chromatin subunits elicit species-specific antibodies against nucleoprotein antigenic determinants.

Nucleosomes composed of 195 base pairs of DNA associated with histones H2A, H2B, H3, and H4 purified from chicken erythrocyte nuclei were used to elicit antibodies in rabbits. Specific serological reaction between the antisera and the nucleosomes is demonstrated by immunodiffusion, immunofluorescence, microcomplement fixation, solid-phase radioimmunoassay, immunosedimentation, and polyacrylamide gel electrophoresis of 5'-32P end-labeled nucleosomes. The antisera did not react with DNA extracted from these nucleosomes, core histones, or the cross-linked histone octamer from chicken erythocytes, calf thymus total histones, or chromosomal proteins HMG-1 or HMG-17. Nucleosome antigenicity was not affected by redigestion with micrococcal nuclease. Digestion with DNase I brought about 50% loss of reactivity while digestion with trypsin or proteinase K resulted in total loss of activity. The antisera reacted strongly with trimer, dimer, and monomer nucleosomes as well as with the core particle (145 base pairs of DNA) and subnucleosome (greater than 145 base pairs) obtained from chicken. It reacted less well with nucleosomes obtained from HeLa cells and was almost totally devoid of activity against chromatin particles obtained from rat liver or wheat germ. Experiments employing the technique of transferring proteins from a polyacrylamide gel to diazobenzyloxymethyl paper and visualization of antigens by autoradiography excluded the possibility that the serum contains antibodies against tissue-specific antigens which are found in small amounts but are very immunogenic. It is concluded that most of the anitbodies in the sera are directed against nucleoprotein antigenic determinants composed of the N-terminal portion of the histones and segments of DNA. Antibody binding is dependent on contact between the histone and DNA segments and is independent of the integrity of the entire nucleosome. Thus, certain histone DNA contacts remain intact even though the structure of the nucleosome has been disrupted.

Solid phase radioimmunoassay for chromosomal components.

This manuscript describes the use of a solid phase radioimmunoassay for serological analysis of chromosomal components. The applicability of this assay for various studies on nonhistone chromosomal proteins, histones, and chromatin subunits is illustrated. By this technique it is possible to detect and quantify nuclear antigens in the nanogram range. The assay has all the inherent sensitivity and precision of radioimmunoassays and, as such, introduces a new, convenient method for serological analyses of chromosomal components. The results presented reconfirm the serological similarity among the HMG (high mobility group) proteins derived from various sources. The amount of HMG proteins present in mononucleosomes purified from calf thymus is similar to that present in mononucleosomes purified from HeLa cells, suggesting that various tissues contain similar amounts of these proteins. Per nucleosome, dinucleosomes and trinucleosomes contain as much HMG-1 protein as mononucleosomes, suggesting that the protein is not exclusively associated with those regions of DNA which have been solubilized by micrococcal nuclease. Part of the antigenic determinants present in HMG-1 forming a complex in the nucleosomal conformation do not interact with antibodies.