Development of somatic embryos from cell suspension cultures of Norway spruce (Picea abies Karst.)

Cell suspension cultures were established from embryonal-suspensor masses derived from mature seeds. Transfer of cell masses on to a medium containing abscisic acid stimulated development of already established individual embryos. Somatic embryos developed shoots when supported by cheese cloth in liquid medium in Petri dishes. The percentage of well-developed roots remained low even though all embryos had root meristems. We have recovered an average of 25 plantlets from an initial PCV of ca 1 g fresh wt per 10 ml.     

Expression cloning of a receptor for C5a anaphylatoxin on differentiated HL-60 cells

A cDNA clone encoding the human C5a anaphylatoxin receptor has been isolated by expression cloning from a CDM8 expression library prepared from mRNA of human myeloid HL-60 cells differentiated to the granulocyte phenotype with dibutyryladenosine cyclic monophosphate. The cDNA clone was able to transfer to COS-7 cells the capacity to specifically bind iodinated human recombinant C5a. The cDNA was 2.3 kb long, with an open reading frame encoding a 350-residue polypeptide. Cross-linking of iodinated C5a to the plasma membrane of transfected COS cells revealed a complex with an apparent molecular mass of 52-55 kDa, similar to that observed for the constitutively expressed receptor in differentiated HL-60 cells or human neutrophils. Although differentiated HL-60 cells display a single class of binding sites, with a dissociation constant of approximately 800-900 pM, the C5a-R cDNA, expressed in COS cells, generates both high-affinity (1.7 nM) and low-affinity (20-25 nM) receptors. Sequence comparison established that the degree of sequence identity between the C5a receptor and the N-formylpeptide receptor is 34%.     

Primary culture of gill epithelial cells from the sea bassDicentrarchus Labrax

Summary We have developed the first explant technique that allows the in vitro study of gill physiology and biochemistry in marine species. Gill fragments were cultured at 17° C, in atmospheric Pco₂, with nutrient medium (Leibovitz L15), pH 7.8, supplemented with 10% fetal bovine serum and adjusted to the osmolarity of fish plasma (350 mOsm/liter). Coating plates with collagen, gelatin, or polylysin did not improve our results. Decrease in osmotic pressure, removal of bovine serum, or its replacement by fish serum inhibited growth from the explants. Approximately 50% of the explants produced cell growth, and after 4 days of culture a monolayer of contiguous cells was formed. This technique is rapid and does not require the use of enzymes. The cells appeared flat and thin with an epithelioid shape. They looked polygonal with a maximum length of 10 to 50 μm. Evidence that they are unique gill cells is the presence of polymorphic surface crenelations (microplicae), prominent Golgi apparatus, tight junctions and desmosomes. Comparison with in vivo tissue showed them to be epithelial cells having differentiated in a homogeneous population of respiratorylike (pavement) cells. They are polarized with their apical surface facing the culture medium. The development of this culture system represents a new tool for cellular approaches to determine precisely the functions and transport mechanism of gill cells.     

Non-exercise daily energy expenditure and physical activity pattern in male endurance athletes.

The present study was performed to determine whether differences in non-exercise daily energy expenditure (Md,ne) exist between trained and untrained individuals. The data from seven cross-country skiers were compared with those from eight sedentary men. Daily energy expenditure (Md) was determined using the heart rate-oxygen consumption relationship; resting metabolic rate (Mr) was measured using indirect calorimetry. A physical activity questionnaire and ratios of Md or Md,ne to Mr were used as indices of physical activity. Md and Mr were significantly higher in the trained subjects whereas Md,ne was identical in the two groups. The ratio of Md,ne to Mr and the data from the physical activity questionnaire showed that there was no significant difference in daily energy expenditure and physical activity pattern during the non-exercise time. These results suggest that the exercise-induced increase in daily energy requirements is not compensated by a more sedentary life during the other daily activities in these trained men.     

Identification des Régimes Alimentaires de la Larve D'un Insecte EntomophagePhanerotoma Flavitestacea [Hym.: Braconidae]

Résumé Les régimes alimentaires d'un insecte parasite entomophage,Phanerotoma flavitestacea, sont identifiés par examen histologique du contenu du mésenteron, au cours de ses 2 modes de vie larvaire successifs, endoparasite, puis ectoparasite. Lors de sa phase endophage, la larve se nourrit uniquement de l'hémolymphe des chenilles d'Anagasta kuehniella. Il n'y a pas de phase stéatophage. Au cours de sa phase ectophage, elle ingère tous les organes de son h?te, sans ordre apparent et sans choix. La digestion, commencée dès le repas achevé, est complètement terminée lorsque le cocon est tissé. L'existence de ces 2 phases et les régimes alimentaires du parasite sont discutés par référence à ce qui est connu chez d'autres Hyménoptères entomophages.

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Summary The alimentary diets of an entomophagous insect larva,Phanerotoma flavitestacea are identified by an histological examination of the mid-gut content, during its two modes of larval life: first endoparasite and then ectoparasite. In the phase as an endophagous larva, it feeds only on the haemolymph ofAnagasta kuehniella caterpillars. During its ectophagous phase, all the internal organs of the host are devoured without choice as well as without following any order in feeding. The digestion begins just after the end of feeding and it is completely finished when the cocoon is spun. The existence of these two phases and the alimentary diets of this parasite are discussed by refering to these phenomena already known in other entomophagous hymenoptera.

     

The phospholipid requirement of the (Ca + Mg)-ATPase from human platelets

The phospholipid requirement of the (Ca²⁺ + Mg²⁺)-ATPase present in a membrane fraction from human platelets was studied using various purified phospholipases. Only those phospholipases, which hydrolyse the negatively charged phospholipids, inhibited the (Ca²⁺ + Mg²⁺)-ATPase activity. The ATPase activity could be restored by adding mixed micelles of Triton X-100 and phosphatidylserine or phosphatidylinositol. Micelles with phosphatidic acid, phosphatidylcholine, phosphatidylethanolamine or sphingomyelin could not be used for reconstitution and inhibited the activity of the native enzyme.     

Chick embryo development can be irreversibly altered by early exposure to weak extremely-low-frequency magnetic fields

Several reports have shown that weak, extremely-low-frequency (ELF), pulsed magnetic fields (PMFs) can adversely affect the early embryonic development of the chick. In this study, freshly fertilized chicken eggs were exposed during the first 48 h of postlaying incubation to PMFs with 100 Hz repetition rate, 1.0 μT peak-to-peak amplitude, and 500 μs pulse duration. Two different pulse waveforms were used, having rise and fall times of 85 μs (PMF-A) or 2.1 μs (PMF-B). It has been reported that, with 2 day exposure, these fields significantly increase the proportion of developmental abnormalities. In the present study, following exposure, the eggs were allowed to incubate for an additional 9 days in the absence of the PMFs. The embryos were taken out of the eggs and studied blind. Each of the two PMF-exposed groups showed an excess in the percentage of developmental anomalies compared with the respective sham-exposed samples. This excess of anomalies was not significant for the PMF-A-treated embryos (P = 0.173), whereas it was significant for the PMF-B-exposed group (P = 0.007), which showed a particularly high rate of early embryonic death. These results reveal that PMFs can induce irreversible developmental alterations and confirm that the pulse waveform can be a determinant factor in the embryonic response to ELF magnetic fields. The data also validate previous work based on the study of PMFs' effects at day 2 of embryonic development under field exposure. © 1994 Wiley-Liss, Inc.