Anion Channel Blockers Inhibit the Acrosome Reaction of Echinoderm Sperm

Two types of anion channel blockers, SITS (4-acetamide-4'-isothiocyanostilbene-2,2'-disulfonic acid) and DIDS (4,4'-diisothiocyanostilbene-2, 2'-disulfonic acid), inhibited jelly-induced acrosorne reaction in starfish and sea urchin. In starfish sperm, both of the blockers reversibly inhibited the formation of acrosomal process but they had no effect on either the acrosomal exocytosis or acid release from the sperm. Complete acrosome reaction occurred even in Cl⁻- and SO₄²⁻-free artificial seawater whereas HCO₃⁻was required for the acrosomal exocytosis. Importance of anion transport in acrosome reaction is discussed.     

Metabolism of Pyrimidines in Protoplasts from Cultured Catharanthus roseus Cells

The metabolism of [2-¹⁴C]thymine, [2-¹⁴C]thymidine, [2-¹⁴C]uraciland [¹⁴C]uridine was investigated in protoplasts obtained fromsuspension cultures of Catharanthus roseus. Most of the exogenouslysupplied thymine, thymidine and uracil was degraded, and salvageof these pyrimidines accounted for 5–36 per cent of thetotal amount of ¹⁴C-labelled precursors which was metabolized.However, more than 80 per cent of the labelled uridine was utilizedfor the biosynthesis of nucleotides and nucleic acids, and therest was degraded. In contrast to the results from protoplastsof sugar cane cells in suspension culture, the data indicatethat protoplasts possess a pathway for the degradation of pyrimidines,and that the overall metabolism of these pyrimidines in protoplastsis very similar to the metabolism in the intact cells. Catharanthus roseus, madagascar periwinkle, protoplasts, pyrimidine metabolism     

Activities of Enzymes Relating to Starch Synthesis and Endogenous Levels of Growth Regulators in Potato Stolon Tips During Tuberization

Changes in contents of starch and protein, and activities of enzymes involved in starch synthesis were studied during tubcrization of stolon tips of Solanum tuberosum L. cv. Irish Cobbler. Starch content and activities of phosphorylase and granule-bound starch synthctase based on fresh weight increased rapidly in the early phase (stage I, the stolon tips just before swelling; stage 2, the swelling tips; stage 3, young tubers of 0.2–0.5 cm diameter), and they all remained nearly unchanged in the later phase (stage 3 to stage 6, young tubers of 3.5 cm diameter). The content of soluble protein based on fresh weight remained unchanged. Activities of soluble starch snythetase and ADP-glucose pyrophosphorylase were not detected at stage 1 and 2, but increased at later stages. Endogenous levels of auxin, cytokinin and gibberellin were assayed for the materials at the corresponding developmental stages. Auxin content was high at stages 1 and 2, and lowered at later stages. Cytokinin content increased abruptly at stage 6. Gibberellin content was low at all stages. The internal conditions for starch deposition and tuberization in potato were discussed in regard to regulation of enzyme activities by growth regulators.     

Influence of Several Growth Regulators and Amino Acids on in vitro Organogenesis of Torenia fournieri Lind.

The effects of several growth regulators and amino acids onin vitro organogenesis of Torenia fournieri Lind. were determinedusing internodal segments. Treatment with 2,4-D¹ resulted innodular callus formation, while NAA and IAA induced roots constantlybut much less frequently shoot buds. Individually BA, zeatin,and 4-PU induced bud formation, but these shoot buds did notdevelop further. Formation of buds by cytokinin was influencedby a simultaneous application of NAA or 2,4-D, but not of IAA,its degree being reduced when BA was simultaneously appliedwith NAA or 2,4-D. When zeatin or kinetin was added with NAA,numerous roots were induced. The effects of various L-amino acids on in vitro organogenesiswere also investigated using the defined medium in which KNO₃was a principal source of nitrogen. The formation of buds wasconsiderably stimulated by alanine and asparagine, and slightlyby glutamic acid in the medium containing both NAA and BA, inwhich bud formation was easily induced. On the other hand, allamino acids except for glutamic acid and aspartic acid inhibitedroom formation in this medium. Root formation was greatly stimulated by proline, alanine, glutamine,glutamic acid, and aspartic acid, and slightly by arginine andtryptophan in the medium containing NAA but no BA. Glutamicacid and aspartic acid also enhanced bud formation in this medium.     

Tissue Distribution of an Immunogenic Melanoma Associated Antigen,D-1

A human melanoma-associated antigen immunogenic in patients was recently identified by screening an expression cDNA library constructed from cultured human melanoma cell line with sera from patients with melanoma. The nucleic acid sequence of the cloned D-1 cDNA has no significant homology with previously reported mammalian genes. The cDNA D-1 encodes a peptide of about 37 kDa, which showed fivefold higher reactivity with sera from patients with melanoma than with sera from normal donors. In order to detect D-1 gene expression in vivo, in-situ hybridization and immunostaining with cRNA probe and murine anti-D-1 sera were carried out on surgically removed tissues. Digoxigenin-labeled cRNA D-1 was exclusively hybridized with mRNA in the cytoplasm of melanoma cells but not with keratinocytes and fibroblasts adjacent to melanoma nests. Polyclonal anti-D-1 antibodies were obtained by immunization of Balb/c mice with recombinant D-1 peptide and clearly reacted with melanoma cells but not with keratinocytes and fibroblasts, similar to the results of in-situ hybridization. The above information will help to assess the suitability of recombinant D-1 peptide to implement active specific immunotherapy in patients with melanoma.     

EFFECTS OF CULTURE CONDITIONS ON HYDROGENASE ACTIVITY OF SCENEDESMUS D3

The effects of culture conditions and of culture ages upon hydrogenaseactivity in Scenedesmus D₃ was investigated with nitrite andbenzoquinone as acceptors. It was noticed that level of ironin the culture medium played an important part in the developmentof hydrogenase activity. The cells grown on a medium of relatively high level of iron(10^–4 M) showed higher activity both in nitrite and benzoquinonereduction than those grown on a medium of relatively low levelof iron (10^–5 M). In the former case, hydrogenase activityappeared even after a short incubation with hydrogen (30 min). When the cells with low hydrogenase activity were transferredto a medium containing 10^–;4 M iron and the activity wasmeasured every 24 hours, the following facts were observed. (a) Nitrite reduction activity increased markedly during thefirst 24 hours and then decreased gradually during further cultivation.Furthermore, a parallelism was observed between nitrite reductionactivity and cell size. (b) On the other hand, the pattern of change in quinone reductionactivity was somewhat different from that of nitrite reduction.This activity appeared only after 3 days' cultivation, reacheda maximum at 4th or 5th day and then decreased. However, thepresence of glucose in the culture medium suppresses the appearanceof quinone reduction activity, while the nitrite reduction activitywas less markedly affected. ¹Present address: Department of Experimental Radiology, Facultyof Medicine, Kyushu University, Fukuoka.     

ROLE OF CALCIUM IN POTASSIUM UPTAKE BY PLANT ROOTS

The effects of EDTA·Mg or GEDTA·Mg on the uptakeof nutrient ions, the release of Ca⁺⁺ and nucleotides into themedium and the nucleic acid contents in rice and red bean rootswere investigated.

1. Both EDTA and EDTA·Mg induced similarly the release ofCa from roots.
2. EDTA·Mg as well as EDTA brought abouta significant repressionin K uptake, but had an insignificantor no effect on P, NH₄and NO₃ uptakes. EDTA·Ca did notrepress K, P, NH₄ andNO₃ uptakes.
3. EDTA or GEDTA acceleratedthe degradation of nucleic acid andthe release of nucleotides,while EDTA·Mg or GEDTA·Mghad no such effect.

These results indicate that the indispensable role of intracellularCa in K uptake does not appear to be directly associated withRNA metabolism. (Received July 29, 1965; )     

Respiratory Utilization of 13C-Labelled Photosynthate in Nodulated Root Systems of Soybean Plants

Kouchi, H., Akao, S. and Yoneyama, T. 1986. Respiratory utilizationof ¹³C-labelled photosynthate in nodulated root systems of soybeanplants.—J. exp. Bot. 37: 985–993. An improved method for the measurement of respiratory utilizationof current photosynthate in the nodulated root system of water-culturedsoybean (Glycine max L.) plants was developed using a steady-state¹³CO₂ labelling technique. Well-nodulated plants at the latevegetative stage were allowed to assimilate ¹³CO₂ for 10 h incontinuous light at a constant CO₂ concentration with a constant¹³C abundance. The respiratory evolution of ¹³CO₂ from rootsand nodules was measured continuously throughout the periodof ¹³CO₂ assimilation and during a subsequent 36 h chase periodby using a differential infrared ¹³CO₂ analyser. The plantswere grown with nitrogen-free or (15 mmol dm^–3)-containing culture solution for 3 d before¹³CO₂ assimilation. In plants grown without , nodule respiration averaged 69% of the total respiration of the undergroundparts over the full experimental period and the CO₂ respiredreached an apparent isotopic equilibrium at 80–85% labellingafter initiating ¹³CO₂ assimilation. By contrast, the CO₂ respiredfrom the roots did not reach an isotopic equilibrium and labellingwas only 56% at the end of exposure to ¹³CO₂ These findingsdemonstrated that nodule respiration is strongly dependent onrecently assimilated carbon compared with root respiration. Plants supplied with in the culture solution showed a decreased rate of nodule respirationand a slightly increased rate of root respiration. The extentsand time courses of labelling of respired CO₂ from both theroots and nodules were similar in the presence and absence of except that the maximum level of labelling of CO₂ derived from nodule respiration in plantswith was significantly higher (about 91%) than for plants growing without . Key words: Soybean (Glycine max L.), nodule respiration, ¹³CO₂, assimilation, carbon partitioning     

A Thermostable Acid α-Glucosidase from Tetrahymena thermophila: Purification and Characterization

An acid α-glucosidase (EC 3.2.1.20) was purified to homogeneity from the culture medium of Tetrahymena thermophila CU 399. Its general molecular, catalytic and immunological properties were compared to those of the T. pyriformis W enzyme. The enzyme from T. thermophila was a 105-kD monomer and the N-terminus (25 amino acid residues) displayed some homology with that of T. pyriformis enzyme. The purified enzyme was most active at 56° C and showed resistance to thermal inactivation. The acid α-glucosidase appears to have α-1,6-glucosidase as well as α-1,4-glucosidase activity. The Km values determined with p-nitrophenyl-α-glucopyranoside, maltose, isomaltose and glycogen were 0.7 mM, 2.5 mM, 28.5 mM and 18.5 mg/ml, respectively. The enzyme was antigenically distinct from T. pyriformis acid α-glucosidase.