Effect of cooling rate,cryoprotectant and holding time at different transfer temperatures on the survival of cryopreserved cell suspension culture (Puccinellia distans (L.) Parl.)

Reflexed saltmarsh-grass suspension cultures produced by seed callus were frozen to the liquid nitrogen temperature. Cooling rates, cryoprotectants and holding times were taken as a function of transfer temperatures. The highest survival of cells (45%) was found at a freezing rate of 1°C min^-1, without cryoprotectant treatments. The cryoprotectants (proline, dimethyl sulphoxide, glycerol), used at different concentrations and transfer temperatures, increased the survival rate. The maximum value was 78% at 12.5% (w/v) of proline with –30°C transfer temperature. Considerable improvement of viability (from 0% to 95%) among the 12.5 and 15.0% (v/v) dimethyl sulphoxide cryopreserved cells was achieved by holding them at – 20°C for 10–30 min before plunging into the liquid nitrogen. A 20 min holding time at 15.0% (v/v) glycerol level and – 30°C transfer temperature significantly enhanced the viability of the explants from 42% to 92%. Plants were successfully regenerated from cells cryopreserved with proline (w/v) and dimethyl sulfoxide (v/v) levels of 12.5 and 15.0%, respectively.     

Consequences of the iron-dependent formation of ferredoxin and flavodoxin on photosynthesis and nitrogen fixation on Anabaena strains

Iron-dependent formation of ferredoxin and flavodoxin was determined in Anabaena ATCC 29413 and ATCC 29211 by a FPLC procedure. In the first species ferredoxin is replaced by flavodoxin at low iron levels in the vegetative cells only. In the heterocysts from Anabaena ATCC 29151, however, flavodoxin is constitutively formed regardless of the iron supply.Replacement of ferredoxin by flavodoxin had no effect on photosynthetic electron transport, whereas nitrogen fixation was decreased under low iron conditions. As ferredoxin and flavodoxin exhibited the same K_m values as electron donors to nitrogenase, an iron-limited synthesis of active nitrogenase was assumed as the reason for inhibited nitrogen fixation. Anabaena ATCC 29211 generally lacks the potential to synthesize flavodoxin. Under iron-starvation conditions, ferredoxin synthesis is limited, with a negative effect on photosynthetic oxygen evolution.     

Daily changes of neuropeptide Y-like immunoreactivity in the suprachiasmatic nucleus of the rat

Most of the biochemical, physiological and behavioural events in living organisms show diurnal fluctuations, normally synchronized with 24-h environmental rhythms, such as the light-dark cycle. The suprachiasmatic nucleus (SCN) of the hypothalamus is considered to be a pacemaker of the circadian rhythms in several mammals. The light-dark cycle is the primary synchronizing agent for many of the circadian rhythms which are regulated by the SCN. The photic information reaches the SCN also through a neuropeptide Y(NPY)-like immunoreactive pathway from the ventro-lateral geniculate nucleus. We found that in 12-h-dark and 12-h-light housed rats the NPY-like immunoreactive innervation of the ventro-lateral part of the SCN shows a 24 h rhythm with values rising gradually during the light phase and falling during the dark phase. Besides this rhythm, we found two peaks corresponding to the switching on and switching off of the light. The average level of NPY-like immunoreactivity, as assessed by means of semiquantitative immunocytochemistry and expressed in 'arbitrary units', is reduced in rats housed in total darkness for 2 weeks. These results confirm the physiological role of NPY in the timing of the circadian activity of the SCN.     

A new species of Lernanthropus De Blainville, 1822 (Copepoda: Lernanthropidae) parasitic on Menticirrhus ophicephalus (Jenyns) (Teleostei: Sciaenidae) from the Peruvian coast

Lernanthropus huamani n. sp. (Copepoda: Lernanthropidae), a parasite of the Peruvian sciaenid fish Menticirrhus ophicephalus (Jenyns), is described and illustrated. The new species differs from all other species of Lernanthropus by a combination of characters, including the dorsal plate, legs and other appendages. L. guacoldae Villalba & Fernández, 1984 is considered a synonym of L. pacificus Oliva & Duran, 1982.     

Progress in botany

Book Review

Progress in botanyH.-D. Behnke, K. Esser, K. Kubitzki, M. Runge and H. Ziegler (Eds.), 50. Berlin: Springer-Verlag, 1989. XIX+386 pages. DM 258.00. ISBN 3-540-50289-0     

Two modes of cell migration in the ventral horn of the spinal cord in the chick embryo. A Golgi study

The migration process of the ventral horn in chick embryo spinal cord cells has been studied between 2.5 and 5 days of incubation (HH-17, HH-26), using the Golgi technique. Two different migratory modes are observed. Type I--Migration by nucleus translocation. Most of the ventral horn motor neurons migrate by nucleus translocation within the peripheral cylinder of the cytoplasm (migration by nucleus translocation). Type II--Free migration cells. Other cells migrate disconnected from both limiting surfaces (ventricular and pial). On the basis of shape and migratory behaviour they have been identified as smooth cells and multipodial cells.     

Ryanodine receptor purified from crayfish skeletal muscle

The ryanodine receptor was isolated from the sarcoplasmic reticulum of crayfish skeletal muscle. Ryanodine binding to the native fraction was measured by Scatchard analysis and values of 60 nmol/l and 9 pmol/mg were obtained for KD and Bmax respectively. The identity of purified receptor was confirmed by electron microscopy, electrophoresis and incorporation into planar lipid bilayers. At least two conductance states (100 pS and 50 pS) were observed in 100 mmol/l NaCl both for native and purified receptor.     

Environmental influences on the sensitization by drugs measured in peripheral blood lymphocyte chromatin

An "allergo-kinetic" method was introduced, which measures early changes of nuclear chromatin structure of lymphocyte subpopulations. The method is used as an in vitro test for drug allergy. The comparison of data from two sampling places (Budapest-Esztergom) shows the following results: The drug allergy scores (obtained from T-cells) may reflect environmental influences on the population under study. During the 13 months period after Atomic Reactor accident in Chernobyl, both the frequency of severe skin manifestations and that of organ manifestations (without skin lesions) increased. The ratio of negative test results decreased in both of drug allergic patient groups (small town = Esztergom, large city = Budapest)--tendency to polysensibilization. The scores obtained in drug related groups of generalized urticaria and Quincke's oedema increased significantly within 13 months after accident followed by a normalization in the subsequent year.     

Multielectrode culture chamber: a device for long-term recording of bioelectric activities in vitro

A multi-microelectrode culture chamber system was constructed for monitoring simultaneously morphological and electrophysiological development of neural cells in vitro. The setup consisted of a pattern of gold conductor lines evaporated onto a glass substrate and insulated with polyamide. The width of each electrode was 10 microns, and the distance between the electrodes was 60 microns. The electrode patterns were constructed and the uncovering of the electrode tips were carried out by photo-etching. This system allowed us to record spontaneous activities in both explant- and primary monolayer cultures of either rat or mouse spinal cords and forebrains, during neuronal regeneration and maturation.