Glycolipid- and glycoprotein-based blood group A antigen expression in human thrombocytes. A1/A2 difference

Total non-acid glycolipid fractions and total sodium dodecylsulphate (SDS) solubilized protein fractions were isolated from human thrombocytes obtained from single human donors having different blood group A₁/A₂ phenotypes. The blood group A glycolipid antigens were characterized by immunostaining of thin layer plates with different monoclonal anti-A antibodies. The glycoproteins carrying blood group A epitopes were identified by SDS-PAGE and Western blot analysis using a monoclonal anti-A antibody. Blood group A glycolipid antigens were found in both A₁ and A₂ thrombocytes but the A₂ individuals expressed at least ten times less A glycolipids compared to the A₁ individuals. Expression of A type 3/4 chain and small amounts of A type 1 chain glycolipids were seen in thrombocytes of both A₁ and A₂ individuals, while the type 2 chain A glycolipids appeared to be missing from the A₂ thrombocytes. Blood group A reactive glycoproteins were only found in thrombocytes of A₁ individuals and could not be detected in A₂ individuals or a blood group O individual. The major blood group A glycoprotein were found as a double band migrating in the 130 kDa region.Abbreviations SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - HPTLC high performance thin layer chromatography - CBB Coomassie brilliant blue - GVH graft versus host Part of this work was presented at the Xth International Symposium on Glycoconjugates, Jerusalem, Israel. September, 1989.In the short hand designation for glycolipids, the letter indicate blood group determinant, the first numeral, the number of sugar residues, and the second numeral, the type of carbohydrate chain. Thus, A-6-1 means a hexaglycosylceramide with a blood group A determinant based on the type 1 carbohydrate chain.     

Proteolysis of ribulose-1,5-bisphosphate carboxylase/oxygenase in leaves of Citrus explants throughout the annual cycle and its regulation by ethylene

Leaf senescence and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBP carboxylase, EC 4.1.1.39) degradation in orange [ Citrus sinensis (L.) Osbeck cv. Washington Navel] explants have been investigated. Explants consisted of a segment of stem (ca 15 cm) and 5 mature leaves. In vitro RuBP carboxylase degradation was determined by culturing the explants in water for different periods of time (3 days usually) and quantifying the two RuBP carboxylase subunits in the extracts following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In vitro RuBP carboxylase degradation was estimated by autodigestion of leaf extracts and SDS-PAGE. The extent of in vivo RuBP carboxylase degradation in explants cultured under 16 h light/8 h dark photoperiod varied throughout the year and showed a cyclic behaviour correlated with the growth cycle of Citrus. The highest proteolytic activity both in vivo and in vitro was found in explants made from April to August coinciding with the maximum vegetative growth period of the tree.
Leaf senescence and abscission could be retarded significantly at any time of the year by maintaining the explants continuously in the dark. Treatment of the explants in the dark with a continuous flow of ethylene enhanced both leaf abscission and rate of RuBP carboxylase degradation, proportionally to ethylene concentration (0.1-0.6 ppm). Ethylene-induced senescence of Citrus leaf explants in the dark appears to be a convenient model system to study the regulation of the proteolytic degradation of RuBP carboxylase.     

Nucleic acid synthesis inChenopodium rubrum L. during photoperiodic induction and its relation to endogenous rhythmicity

Beginning with the second inductive cycle the rate of nucleic acid (NA) synthesis in cotyledons and apical buds ofChenopodium rubrum is higher at the end of the dark period or 4h following transfer of the plants to light in induced plants than in non-induced ones. This is due to an increase in all NA fractions. The greatest difference between NA synthesis in induced and non-induced plants was observed at the end of the second (or sometimes third) inductivecycle. In the subsequent cycles the difference decreased or disappeared eventually. During photoperiodic induction NA synthesis shows a diurnal rhythm with a peak at the end of the dark and at the beginning of the light period. Rhythmicity of NA synthesis is endogenous. The period length of the endogenous oscillation is about 18 h. Interruption of the dark period by light causea amplitude of the first oscillation to be reduced and delays the appearance of the second peak. NA synthesis did not show rhythmicity in plants grown in continuous light. The significance of the observed phenomena for photoperiodic induction is being discussed.     

Changes in the level of endogenous gibberellins and auxins in apical buds ofChenopodium rubrum L. after application of growth substances reversing the effect of (2-chlorethyl)-trimethylammonium chloride (CCC) on flowering

The application of CCC at concentrations inhibiting flowering ofChenopodium rubrum reduces the level of endogenous gibberellins in the apical buds of the plants. The effect of CCC may be reversed by appropriate concentrations of gibberellin (GA-), indole acetic acid (IAA) or kinetin. Kinetin applied to the apical bud during floral induction reduced the level of endogenous gibberellins similarly as CCC and if both CCC and kinetin were applied simultaneously their action was additive. On the other hand IAA applied under the same conditions increased the level of endogenous gibberellins and after joint application of CCC and IAA their level was the same as in untreated control plants. After application of CCC during floral induction the level of endogenous auxins did not change markedly but an active substance “x” appeared on the chromatograms of indole compounds. This substance was found also after simultaneous application of GA- and CCC but not after joint application of CCC and kinetin. If follows from our results that the same morphological phenomenon (flowering) can take place in plants considerably differing as to their level of endogenous growth substances. The ratio of different growth substances is obviously more important than the actual level of the single substances.     

A high level of hybrid dysgenesis in Drosophila: High thermosensitivity, dependence on DNA repair, and incomplete cytotype regulation

Summary An unusually high level of P-M hybrid dysgenesis in Drosophila melanogaster is characteristic of hybrid offspring originating from both, A (M × ) and B (P × M) crosses of a subline of the Harwich P strain, termed H ^s . The novel properties induced by mobility of P elements carried by H ^s paternal chromosomes include: very high (over 95%) gonadal dysgenesis (GD) in both sexes at the low restrictive temperature of 21°C, and highly premature sterility when males are reared at 18°C and aged at 21°C. Although all three major chromosomes of the H ^s subline contributed to this atypical pattern of gonadal dysgenesis, chromosome 3 had the largest effect. Gonadal dysgenesis showed a temperature- and sex-dependent repression pattern by the defective P elements of Muller-5 Birmingham chromosomes; at 21°C there was virtually no repression of male sterility, but most effective repression of GD in females. At 29°C repression was effective in males, but declined in females. The high thermosensitive sterility, low fecundity, and premature aging of the male germ line were greatly exacerbated when males derived from either A or B crosses were deficient either in excision repair (mei-9 mutation) or in post-replication repair (mei-41 mutation). These findings demonstrate that both DNA repair pathways are essential for the repair of lesions induced by P element transposition and support the hypothesis that P element-induced chromosome breaks are responsible for the virtual abolition of the germ line. The relatively high premature sterility of cross B DNA repair-deficient males, reared at 18°C and aged at 21°C, indicates that there is incomplete cytotype regulation in H ^s subline hybrids.     

Fine structure and functional role of the coverings of the eggs inMesostoma ehrenbergii (Focke) (Turbellaria,Neorhabdocoela)

Summary European populations of the turbellarianMesostoma ehrenbergii can produce two types of eggs according to a seasonal cycle. Subitaneous eggs (S eggs) are produced throughout the warm season and develop rapidly inside the parental uteri. They are meant to ensure a fast increase in the number of worms. Dormant eggs (D eggs) are produced during the warm season and are usually deposited in water at the death of the parent. They can hatch at the beginning of the next warm season and are, therefore, meant to ensure the survival of the population throughout the cold season, when adult worms are no longer present.S eggs are bounded by a thin, translucent covering which has a loose structure and is mainly proteinaceous in composition. These characteristics account for the functional role played by the S egg covering during embryonic development. The covering has, in fact, to be permeable to parental nutrients, and to keep pace, by stretching, with the enlargement of the embryo. By contrast, D eggs are bounded by a thick shell, presumably composed of a sclerotin, the precursors of which (polyphenols) are synthesized together with yolk in the vitelline cells. The shell has a smooth surface devoid of pores and has the function of isolating the developing D egg from the external environment.Supported by a grant from the Consiglio Nazionale della RicercheThe authors wish to thank Dr. P. Roach for his help in translating the paper