Histological and ultrastructural studies of Beauveria bassiana infection in Leptinotarsa decemlineta larvae during ecdysis

Studies of Leptinotarsa decemlineata larvae infected by Beauveria bassiana during ecdysis have enabled us to define the modes of fungal penetration employed to enter the ecdysial cuticle. We have observed the mechanically active passage of the penetrant hyphae and have followed the growth of the filaments and blastospore formation in the molting fluid. The attack of the new integument and its consequent alteration and the entry into the body cavity have also been studied. The infection develops rapidly in some of the larvae which die in premolt, while others are able to molt. Conditions rendered abnormal due to the presence of the fungus cause integumentary injuries which serve as an important factor in pathogenesis since they enhance the entry of fungal elements and bacteria thereby inducing septicemia. Contaminated larvae are able to molt, showing no signs of injury or disease, and survive for a long time, until the fungus finally invades the organism and causes death. This postponement of mortality shows that molting and hemocytic reactions are, to a certain extent, an effective defense mechanism. These last observations can be useful in the understanding of pathological processes associated with a hidden phase of fungal infection.     

Microspatial heterogeneity in the distribution of ciliates in a small pond

Five transects of contiguous samples from the surface of a small pond and one transect from its bottom were collected in order to quantify microspatial heterogeneity in the distribution of ciliated protozoa. Examination of the frequency-abundance relations for these transects suggests that they can be approximated by negative binomial distributions with a commonk of 1.87. Contagiousness or crowding increases with population density.Mean patch size and mean interpatch distance were measured for 4 transects as 1.5 to 2 cm and 3 to 4 cm, respectively. This heterogeneity is suggested to arise from behavioral aggregation about discrete food sources and be very ephemeral.Blocking of adjacent contiguous samples was used to investigate the effect of sample size on the apparent correlation between the numbers of pairs of taxa. In all cases examined, taxa were relatively independent in their distribution at small sample sizes and became more negatively or positively associated as samples were combined. This may reflect that the small scale patches are essentially monospecific.     

Contribution to the structural characterization of eucaryotic PSI reaction centre—I. Critical analysis of the polypeptidic composition of different P700 enriched fractions

In thylakoid membranes, several peptides of high MW are present which may interfere with the study of CP1's components. Modifying Cleveland's technique [7] for limited proteolysis, we have characterized the polypeptides found in the 60 kD region. Some may result from incomplete washing of the CF1 while others come from the CP1; indeed, this chlorophyll protein complex, which has a higher MW (above 100 kD), very often undergoes a dissociation into smaller components of about 60 KD MW.Analysis of the protein content of different preparations commonly used to obtain PSI reaction centre enriched fractions has been performed. The and subunits of CF1 are among the main contaminants of most of these preparations. A further purification step is described which can be applied to all these preparations, but numerous peptides are still present in the active fractions. It is most unlikely that all these polypeptides are required for the primary photochemical event, and this emphasizes the necessity to find a new simple method to purify PSI reaction centres.     

Contribution to the structural characterization of eucaryotic PSI reaction centre — II. Characterization of a highly purified photoactive SDS-CP1 complex

Under precise conditions, SDS PAGE allows purification of a photoactive P700-chla-protein complex from eucaryotic cells. The yield of P700 recovery is close to 100%. A total protein content equivalent to about 140 kD for one mole of P700 has been estimated by chemical analysis, and electrophoresis revealed the presence of two peptidic chains with MWs close to 65 kD. Photochemical and structural properties of this complex are given and compared with those of other complexes previously isolated.     

Orientation of the hemes of high potential cytochromes relative to photosynthetic membranes, as shown by the linear dichroism of oriented preparations

The orientations of high potential cytochromes with respect to photosynthetic membranes was investigated in spinach chloroplasts and in Rhodopseudomonas viridis. The general approach consists in detection with polarized light of photoinduced absorbance changes related to the oxidation of the cytochromes. The orientation of cytochrome c-558 was measured at room temperature in chromatophores and whole cells of Rps. viridis, oriented on glass slides and in a magnetic field, respectively. The orientation of cytochrome b-559 of green plants was detected at 77 K in magnetically oriented chloroplasts. In both cases the dichroic ratio for the band shows that the heme plane makes an angle greater than 35°C with the membrane plane. Moreover, the dichroic ratio is not constant throughout the and β bands, for both cytochrome c-558 and b-559. Linear dichroism spectra of oriented pure horse heart cytochrome c and cytochrome c₂ of Rhodopseudomonas sphaeroides in stretched polyvinyl alcohol films show that the variations of the dichroic ratio in the and β bands can be explained by the occurrence of x- and y-polarized transitions absorbing at slightly different wavelengths.     

Effect of sodium butyrate on the hepatoma cell cycle: Possible use for cell synchronization

Summary Exposure of HTC cells to sodium butyrate caused inhibition of growth. The site of growth inhibition was studied by time-lapse cinematography and [³H]thymidine incorporation studies. Evidence is presented that sodiunm butyrate affected the cell cycle at a specific point immediately after mitosis. Inasmuch as it does not modify the interphase duration after its removal, butyrate may be used for HTC synchronization. This work was supported by l'Institut Nationale de la Santé et de la Recherche Médicale and la Centre Nationale de la Recherche Scientifique (L. T. and J. K.).     

Ultrastructural modifications of the glandular epithelium of the receptaculum seminis in Thermobia domestica (Insecta: Thysanura) during the moulting period

Summary The wall of the receptaculum seminis of Thermobia domestica is composed of numerous glandular units, each with four enveloping cells (denoted 1 to 4) separated by ordinary epithelial cells and associated with a cuticular apparatus. During the moulting periods, which continue to occur in the adult stage, these cells undergo a series of transformations. Just before apolysis there is a dedifferentiation of numerous cytoplasmic organelles, but no mitosis has been observed. When the intima lifts off, the apical system of each glandular unit, i.e. the distal parts of the C2 and C3 cells surrounding the end apparatus, is also eliminated. Then at the apex of each glandular unit, a new ductule is formed in the cavity of which a long ciliary process grows up from cell C1. Finally comes the phase of cuticle formation, i.e., epicuticle for the ductules, epi-and endocuticle for the intima lining the central cavity of the receptaculum. Various cell types participate in secretion of cuticle, the ciliary cells (C1) being responsible for the formation of the porous end apparatus. At ecdysis almost all of the new intima has been secreted and the apical systems are once more differentiated. These transformations are compared with those recently described in other exocrine glands of arthropods, e.g., tegumentary glands and accessory glands of the genital ducts.     

Studies on the mechanism of NADPH oxidation by the granule fraction isolated from human resting polymorphonuclear blood cells

Various factor affecting NADPH-oxidation by resting human leucocyte granules (LG) at acid pH, have been investigated.It was found that:

1) oxidation of NADPH by LG was increasingly inhibited by increased cyanide concentrations in the medium and was abolished by 4 mM cyanide.

2) with or without cyanide in the incubation medium, LG omitted, Mn⁺⁺, in the presence of NADPH induced superoxide anion (O¯₂) production, as evidenced by oxygen consumption and H₂O₂ production, which were abolished (in the absence of cyanide) by cytochrome C (a potent O¯₂ scavenger).

3) Both NADPH oxidation in the presence of 2 mM cyanide (cyanide-resistant) and in its absence (cyanide-sensitive) by LG occured only in the presence of Mn⁺⁺, and both were inhibited by superoxide dismutase.

4) Cyanide-resistant NADPH oxidation by LG generated H₂O₂, was inhibited by H₂O₂ and was not modified by «active catalase. The ratio of cyanide-resistant NADPH oxidation/O₂ uptake was 1 up to 1.25 mM NADPH, and increased above this concentration.

5) Cyanide-sensitive NADPH oxidation was inhibited by catalase and increased upon addition of H₂O₂. The ratio of cyanide-sensitive NADPH oxidation/O₂ uptake was 2.

It was concluded that after initiation by O¯₂, produced independently of LG, two sequential types of LG dependent NADPH oxidations occur. First, an O¯₂-dependent protein mediated NADPH oxidation (cyanide-resistant) which generates H₂O₂ and O¯₂ occurs. Second, NADPH peroxidation (cyanide-sensitive) which utilizes H₂O₂ takes place.     

Partial 9q trisomy associated with a 9,21 translocation

Summary A new case of partial trisomy 9q was found in a child presenting two de novo aberrations: a deletion of the long arms of 9 and a 9,21 translocation. A tentative cytogenetic explanation is put forward.