Different effects of concentric and eccentric muscle actions on plasma volume

The effects of concentric (CON) and eccentric (ECC) contractions on Delta plasma volume (PV), heart rate (HR), and lactate in responses to protocols in different body positions were investigated. CON or ECC contractions were performed in either a single-exercise (6 sets of 12 repetitions of leg extensions completed at 80% of 12 repetition maximum [12RM] with 3-minute rest periods) or multiexercise (4 sets of 10 repetitions for both CON and ECC trials of bench press, leg extension, military press, and leg curl at 80% of 10RM with 90-second rest periods) protocols. HR and lactate increased significantly for both protocols from pre- to postexercise for CON but not ECC trials. DeltaPV was greater following both CON single-exercise (-11.48 +/- 1.38%) and multiexercise (-4.64 +/- 0.33%) trials vs. ECC single-exercise (-1.62 +/- 1.69%) and multiexercise (-1.26 +/- 1.20) trials. Data demonstrate ECC exercise in response to single and multiexercise protocols at the same absolute workload as CON exercise produces less cardiovascular stress.     

Development of a biosensor for on-line detection of tributyltin with a recombinant bioluminescent Escherichia coli strain

A biosensor was developed for the detection of tributyltin (TBT), using a bioluminescent recombinant Escherichia coli:: luxAB strain. Dedicated devices allowed the on-line measurement of bioluminescence, pH and dissolved oxygen values and the feed-back regulation of temperature. Bacterial physiology was monitored by the measurement of the cellular density, respiratory activity and the intracellular level of ATP, glucose and acetate levels. Our results showed that a synthetic glucose medium gave a better TBT detection limit than LB medium (respectively 0.02 micro M and 1.5 micro M TBT). High growth and dilution rates ( D=0.9 h(-1)) allowed maximum light emission from the bacterium. Moreover, simple atmospheric air bubbling was sufficient to provide oxygen for growth and the bioluminescence reaction. Real-time monitoring of bioluminescence after TBT induction occurred with continuous addition of decanal up to 300 micro M, which was not toxic throughout a 7-day experiment. The design of our biosensor and the optimization of the main parameters that influence microbial activity led to the capacity for the detection of TBT.     

Depletion of the photosystem II core complex in mature tobacco leaves infected by the flavum strain of tobacco mosaic virus

The flavum strain of Tobacco mosaic virus (TMV) differs from the wild-type (wt) virus by causing strong yellow and green mosaic in the systemically infected developing leaves, yellowing in the fully expanded leaves, and distinct malformations of chloroplasts in both types of infected tissues. Analysis of the thylakoid proteins of flavum strain-infected tobacco leaves indicated that the chlorosis in mature leaves was accompanied by depletion of the entire photosystem II (PSII) core complexes and the 33-kDa protein of the oxygen evolving complex. The only change observed in the thylakoid proteins of the corresponding wt TMV-infected leaves was a slight reduction of the alpha and beta subunits of the ATP synthase complex. The coat proteins of different yellowing strains of TMV are known to effectively accumulate inside chloroplasts, but in this work, the viral movement protein also was detected in association with the thylakoid membranes of flavum strain-infected leaves. The mRNAs of different enzymes involved in the chlorophyll biosynthesis pathway were not reduced in the mature chlorotic leaves. These results suggest that the chlorosis was not caused by reduction of pigment biosynthesis, but rather, by reduction of specific proteins of the PSII core complexes and by consequent break-down of the pigments.     

Developmental changes in cyclin B1 and cyclin-dependent kinase 1 (CDK1) levels in the different populations of spermatogenic cells of the post-natal rat testis

Spermatogenesis is a highly ordered process which requires mitotic and meiotic divisions. In this work, we studied the relative changes in the levels of the two components of the M-phase promoting factor (MPF): the regulatory subunit cyclin B1 (CycB1) and its catalytic subunit cdk1, in spermatogenic cells of rats between 16 and 90 days of life. A multivariate flow cytometry analysis of forward scatter (FSC), side scatter (SSC) and DNA content was used to identify six populations of rat germ cells: spermatogonia with preleptotene spermatocytes, young pachytene spermatocytes, middle to late pachytene spermatocytes, secondary spermatocytes with doublets of round spermatids, round spermatids, and elongated spermatids. For any population studied no significant difference in the relative cellular content of CycB1 or cdk1 proteins between animals of different ages was observed. By contrast, CycB1 and cdk1 levels were different between the different populations of germ cells. CycB1 and cdk1 were rather high in young pachytene spermatocytes and culminated in late spermatocytes, i.e. just before the first meiotic division. The relative levels of the two proteins remained high in secondary spermatocytes then decreased in round spermatids at the exit of meiosis. Similar results were obtained by Western-blot analysis of total proteins obtained from lysates of elutriated fractions of spermatocytes and spermatids. MPF activity was assessed in lysates of germ cells from 32-day-old rats or adult animals using p13suc1 agarose and histone H1 as an exogenous substrate. H1 kinase activity was higher in pachytene spermatocytes than in round spermatid fractions from both adult and young rats. These results indicate that the meiotic G2/M transition is associated to high levels of CycB1 and cdk1 leading to high MPF activity irrespective of the age of the animals.     

Microgravity and hypergravity effects on fertilization of the salamander Pleurodeles waltl (urodele amphibian)

Effects of microgravity (microG) on fertilization were studied in the urodele amphibian Pleurodeles waltl on board the MIR space station. Genetic and cytomorphologic analyses ruled out parthenogenesis or gynogenesis and proved that fertilization did occur in microG. Actual fertilization was demonstrated by the analysis of the distribution of peptidase-1 genes, a polymorphic sex-linked enzyme, in progenies obtained in microG. Further evidence of fertilization was provided by the presence of spermatozoa in the perivitelline space and in the fertilization layer of the microG eggs and by the presence of a female pronucleus and male pronuclei in the egg cytoplasm. Experiments in microG and in 1.4G, 2G, and 3G hypergravity showed for the first time that, compared to eggs in 1G, several characteristics of the fertilization process including the cortical reaction and the microvillus transformations were altered depending on the gravitational force applied to the eggs. Microvillus elevation, the most evident feature, was reduced on microG-eggs and amplified on eggs submitted to 2G and 3G. No lethal consequences of these alterations on the early development of microG-eggs were observed.     

Nitrogen excretion and changes in blood components during emersion of the subtidal spider crab Maia squinado (L.)

Survival ability of Maia squinado to emersion and subsequent reimmersion was determined in winter and summer conditions. Male spider crabs were less tolerant of emersion than females. Emersion (up to 24 h in summer and to 48 h in winter) induced a marked reduction of nitrogen excretion, especially ammonia excretion. Increase in blood ammonia content was rapid and very high in summer (1750 micromol l(-1)), but non-lethal levels. Estimation of the body ammonia overload showed that only 30% of unexcreted ammonia accumulated in blood. The ammonia release at reimmersion indicated that ammonia also accumulated in other body compartments. Increase in blood urate content, which indirectly reduces ammonia production, was similar at both seasons. Emersed M. squinado was rapidly resorting to anaerobic metabolism, especially in summer when its blood haemocyanin content is low. A strong hyperglycemia was developed in the first 12 h of emersion at both seasons. Mortality occurring beyond 24 h of reimmersion, when the body ammonia overload is cancelled and the recovery of most of blood components is achieved, remains unexplained.     

Spectroscopic studies on ethidium bromide binding to intramolecular parallel and antiparallel triple helices containing T*A:T and G*G:C triplets

The interaction of ethidium bromide (EB), a DNA intercalator, with two intramolecular triplexes 5'd(G4A4G4-[T4]-C4T4C4-[T4]-G4T4G4), 5'd(G4T4G4-[T4]-G4A4G4-[T4]-C4T4C4) ([T4] represents a stretch of 4 thymine residues) and their precursor duplexes has been investigated by circular dichroism, fluorescence and UV absorption spectroscopy. Binding of EB induces a circular dichroism band in the region around 310 nm which is positive for the duplex forms but negative for the triplex forms. We observed that the binding of EB to the duplex form does not induce the formation of the triplex structures. Thermal denaturation experiments demonstrate that EB stabilizes more the parallel triple helix than the antiparallel one. Analysis of the binding process from fluorescence measurements shows that binding constants to the triple helical forms and to the hairpin reference duplex [T4]-G4A4G4-[T4]-C4T4C4) are close. However the binding site size is larger for the triplexes (4-6 base triplets) than for the duplex (2 base pairs).     

Distribution and palaeoecology of freshwater algae and stromatolites: III, some new forms from the Carboniferous, Permian and Triassic of France and Spain

New collections allow us to describe some new algae forming: a) centimetric, unorganized, masses (Toutinella bifurcata from the Middle Triassic of Provence); b) large, decimetric masses (Zarramanella menorica from the ‘Buntsandstein’ of Minorca, Balearic Islands) and c) interesting, plurispecific biocenosis, laminated and silicified, from the French Massif Central: Lower Stephanian of Saint-Étienne basin (with Hankerpella minima, Baltzerella stephaniense, Ponsinella undulata) and Lower Autunian of Bourbon-l'Archambault basin (Hanskerpella pustulata, Baltzerella stephaniense, Ponsinella multifrons, Sarfatigirella intricata). Other forms are badly preserved, one is possibly a Broutinella sp. and another may be a coccoid form of cyanobacteria or chlorophyceae. Silicified biocenoses macroscopically occur in the form of regular laminations, exhibiting vertical, planar joints (1–2 cm wide) and horizontal, planar joints, 5 mm to 3 cm thick, with various sedimentary infillings. These facies represent early and late silicifications of biological felts (mats), probably previously partly calcified and affected by desiccation cracks partly infilled before the silicification. There is no evidence for an early silicification of living filaments and trichomes. The silicification also affects other types of sediments (petites, sandstones, conglomerates, breccias) associated with the algal formations. Macroscopically, these laminated formations do not show domes or hemispheres; on the other hand, these features clearly appear under the microscope, and these formations are true stromatolites. The basins of Saint-Étienne and Bourbon-l'Archambault contain volcanic rocks; the siliceous series, very localized in the basins, represent an impregnation of various primary materials by silica of hydrothermal origin. The present study is voluntarily limited to the palaeontological and palaeoecological appearances; algal laminations formed in ephemeral pools and small lakes, under very particular conditions, ‘hydrothermal’ sensu lato, are still to be stated more precisely.

Résumé

Des récoltes nouvelles permettent de décrire quelques algues nouvelles formant des amas centimétriques non organisés (Toutinella bifurcata du Trias moyen de Provence), de grosses masses décimétriques (Zarmmanella menorica du Buntsandstein de Minorque) et d'intéressantes biocrenoses plurispécifiques laminées et silicifiées du Massif central français: Stephanien inférieur de Saint-Étienne (à Hankerpella minima, Baltzerella stephaniense, Ponsinella undulata), Autunien inférieur du bassin de Bourbon-l'Archambault (Hanskerpella pustulata, Baltzerella stephaniense, Ponxinella multifrons, Sarfatigirella intricata). D'autres formes sont mal conservées, l'une serait attribuables à une ? Broutinella sp. et une autre à une coccoide (cyanobactéries ou chlorophycées). Les biocœnoses silicificées se présentent macroscopiquement sous forme de laminations régulières, accidentées de fentes planes verticales (1–2 cm de largeur) et de fentes planes horizontales (5 mm–3 cm de hauteur), à remplissages sédimentaires variés. Ces faciès correspondent à des silicifications précoces et tardives de feutres biologiques probablement en grande partie calcifiés précocement et affectés de fentes de dessiccation partiellement remplies avant la silicification. Aucun indice de silicification de filaments et de trichomes du vivant des organismes n'a pu être observé. La silicification affecte aussi d'autres sédiments (pélites, grès, conglomérats, brèches), associés aux anciennes formations algaires. Macrospiquement, ces formations laminées ne montrent pas de dômes ni d'hémisphères; en revanche ces structures apparaissent parfaitement au microscope, et ce sont de véritables stromatolites. Les bassins de Saint-Étienne el de Bourbon-l'Archambault contiennent des formations volcaniques; les formations siliceuses, qui sont très localisées dans les bassins, correspondent certainement à une imprégnation par de la silice d'origine hydrothermale de sédiments variés. La présente étude se limite volontairement à l'aspect paléontologique et paléoécologique; les laminations algaires se sont formées dans des bassins ou de petits lacs éphémères, dans des conditions très particulières, hydrothermales sensu lato, encore à préciser.