The modification of the conserved GXXXG motif of the membrane-spanning segment of subunit g destabilizes the supramolecular species of yeast ATP synthase

The supernumerary subunit g is found in all mitochondrial ATP synthases. Most of the conserved amino acid residues are present in the membrane C-terminal part of the protein that contains a dimerization motif GXXXG. In yeast, alteration of this motif leads to the loss of subunit g and of supramolecular structures of the ATP synthase with concomitant appearance of anomalous mitochondrial morphologies. Disulfide bond formation involving an engineered cysteine in position 109 of subunit g and the endogenous cysteine 28 of subunit e promoted g + g, e + g, and e + e adducts, thus revealing the proximity in the mitochondrial membrane of several subunits e and g. Disulfide bond formation between two subunits g in mitochondria increased the stability of an oligomeric structure of the ATP synthase in digitonin extracts. These data suggest the participation of the dimerization motif of subunit g in the formation of supramolecular structures and is in favor of the existence of ATP synthase associations, in the inner mitochondrial membrane, whose masses are higher than those of ATP synthase dimers.     

The response of human colonocytes to folate deficiency in vitro: functional and proteomic analyses

Low folate intake is associated with colon cancer. We combined a proteomics and biochemical approach to identify proteins and pathways affected by folate deficiency in human colonocytes. Folate differentially altered activity and expression of proteins involved in proliferation [e.g., PCNA], DNA repair [e.g., XRCC5, MSH2], apoptosis [e.g., BAG family chaperone protein, DIABLO and porin], cytoskeletal organization [e.g., actin, ezrin, elfin], and expression of proteins implicated in malignant transformation [COMT, Nit2].     

Feeding ecology of juvenile flatfishes of the surf zone of a sandy beach

Prey items of 0- and 1-group plaice Pleuronectes platessa , sole Solea solea , brill Scophthalmus rhombus , turbot S. maximus and dab Limanda limanda of the surf zone of a Belgian sandy beach, included hyperbenthic (e.g. mysids), endobenthic (e.g. polychaetes) and epibenthic (e.g. shrimps) species. Little dietary overlap was observed. If diet overlap did occur, it mainly involved prey species that are dominant in the surf zone of Belgian beaches, such as shrimps and mysids. These results suggest an opportunistic utilization by flatfish of the available food resources in surf zone ecosystems. Also, two strategically different feeding habits could be distinguished between the five flatfish species. Turbot and brill mainly fed on large, highly mobile prey (e.g. fish, mysids) and had a rather narrow prey spectrum, whereas plaice, dab and sole ate more benthic prey (e.g. polychaetes) and had a broader prey spectrum.     

The yeast F(1)F(0)-ATP synthase: analysis of the molecular organization of subunit g and the importance of a conserved GXXXG motif

The F(1)F(0)-ATP synthase enzyme is located in the inner mitochondrial membrane, where it forms dimeric complexes. Dimerization of the ATP synthase involves the physical association of the neighboring membrane-embedded F(0)-sectors. In yeast, the F(0)-sector subunits g and e (Su g and Su e, respectively) play a key role in supporting the formation of ATP synthase dimers. In this study we have focused on Su g to gain a better understanding of the function and the molecular organization of this subunit within the ATP synthase complex. Su g proteins contain a GXXXG motif (G is glycine, and X is any amino acid) in their single transmembrane segment. GXXXG can be a dimerization motif that supports helix-helix interactions between neighboring transmembrane segments. We demonstrate here that the GXXXG motif is important for the function and in particular for the stability of Su g within the ATP synthase. Using site-directed mutagenesis and cross-linking approaches, we demonstrate that Su g and Su e interact, and our findings emphasize the importance of the membrane anchor regions of these proteins for their interaction. Su e also contains a conserved GXXXG motif in its membrane anchor. However, data presented here would suggest that an intact GXXXG motif in Su g is not essential for the Su g-Su e interaction. We suggest that the GXXXG motif may not be the sole basis for a Su g-Su e interaction, and possibly these dimerization motifs may enable both Su g and Su e to interact with another mitochondrial protein.     

Contact relations and guidance of primordial germ cells on their migratory route in embryos of Xenopus laevis

This paper describes the relationship between primordial germ cells (p.g.cs) and the substrate over which they migrate in early embryos of the anuran amphibian Xenopus laevis. P.g.cs migrate from the embryonic gut to the dorsal body wall along the dorsal mesentery at the earliest swimming stage. Our earlier papers have described the way in which p.g.cs move in vitro. In this work we have studied the shape and cytoarchitecture of both p.g.cs and the coelomic epithelial cells (c.e.cs) over which they migrate. We have concentrated on three aspects of the morphology of these cells: first the shapes of the c.e.cs and the way that they affect the shapes of the p.g.cs; secondly the presence of adhesion plaques between the two types of cell; and thirdly the arrangement of cytoskeleton elements. The results show that c.e.cs in the dorsal mesentery are orientated cranio-caudally while those on the dorsal body wall and at the junction with the mesentery are arranged transversely, at 90 degrees to the cranio-caudal plane. P.g.cs are found in both elongated and rounded state. Where elongated, they are always in the same plane as the c.e.cs with which they are associated. The implications of this are discussed. Adhesion plaques between p.g.cs and c.e.cs are shown both by disaggregation studies and transmission electron microscope studies. Plaques are associated with the well defined microfilamentous cytoskeleton of c.e.cs, but only with a sparse array of filaments in p.g.cs. The only parts of p.g.cs where filaments are regularly found are their filopodia, which are generally seen on elongated p.g.cs in longitudinal section. We suggest on the basis of this work that p.g.cs have a dispersed cytoskeleton except during filopod extension, that they move by forming direct adhesion plaques with c.e.cs, and that c.e.cs provide a firm orientated support and possible guide to p.g.c. movement.     

Demineralization in organic solvents by alkylammonium salts of ethylenediaminetetra-acetic acid.

1. Insoluble salts of alklaine earth metals (e. g. calcium phosphate) dissolve in non-aqueous solvents containing EDTA salts of organic cations (e.g. tetraethylammonium). 2. Bone is efficiently demineralized in ethanolic trimethylammonium EDTA. Retention in the matrix of water-soluble materials (e.g. proteoglycan) is much better than in standard aqueous EDTA demineralization procedures.     

Estrogenic and antiestrogenic properties of clomiphene citrate in laboratory mice

The estrogen agonistic and antagonistic properties of clomiphene citrate were investigated in the mice. Clomiphene citrate was tested at various doses of 0.1, 1.0, 10 and 100 μg for three consecutive days in immature and mature bilaterally ovariectomized mice. Clomiphene citrate showed uterotrophic activity in both immature and ovariectomized conditions. The lower doses of 0.1 and 1.0 μg were ineffective to show any uterotrophic stimulation. Clomiphene citrate at 10 μg dose produced 305.56% increase in uterine weight i.e., 27.70 ± 0.24 vs 6.83 ± 0.06 in immature and 182.27% i.e., 42.68 ± 1.12 vs 15.12 ± 0.57 in ovariectomized mice. Clomiphene citrate at 100 μg dose showed significant uterotrophic effect e.g., 435.57% i.e., 36.58 ±0.34 vs 6.83 ± 0.06 in immature and 586% i.e., 103.80 ± 0.60 in ovariectomized mice. When clomiphene citrate was administered in combination with 0.32 μg of estradiol 17-β it caused significant antagonistic effect (decrease in uterine weight) at 10 and 100 μg respectively. Clomiphene citrate at 10 μg dose produced 32% i.e., 28.93 ± 0.43 vs 38.04 ± 2.68 in immature and 35% i.e., 59.64±1.44 vs 83.34 ±0.25 in ovariectomized mice respectively. Histological observation clearly showed that clomiphene citrate at 10 and 100 μg doses did not cause any differential hypertrophy of the epithelial layer. Similar doses in combination with estradiol produced significant antagonistic effect on uterine weight and luminal epithelial cell height.     

The modulation in subunits e and g amounts of yeast ATP synthase modifies mitochondrial cristae morphology

Subunits e and g of Saccharomyces cerevisiae ATP synthase are required to maintain ATP synthase dimeric forms. Mutants devoid of these subunits display anomalous mitochondrial morphologies. An expression system regulated by doxycycline was used to modulate the expression of the genes encoding the subunits e and g. A decrease in the amount of subunit e induces a decrease in the amount of subunit g, but a decrease in the amount of subunit g does not affect subunit e. The loss of subunit e or g leads to the loss of supramolecular structures of ATP synthase, which is fully reversible upon removal of doxycycline. In the absence of doxycycline, mitochondria present poorly defined cristae. In the presence of doxycycline, onion-like structures are formed after five generations. When doxycycline is removed after five generations, cristae are mainly observed. The data demonstrate that the inner structure of mitochondria depends upon the ability of ATP synthase to make supramolecular structures.     

Neighbouring group participation in the unblocking of phosphotriesters of nucleic acids.

Two examples of neighbouring group participation during the removal of protecting groups from phosphotriesters of partially or fully protected intermediates of nucleic acids are presented. The first example shows that ammonolysis of aryl groups from phosphotriesters of partially protected - 5'- hydroxy free - nucleic acids (e.g., 4b approximately to; Ar=2C1C 6H4) gives rise to the formation of unnatural nucleic acids (e.g., 7 approximately to and 8 approximately to). The second one illustrates that fluoride ion promoted hydrolysis of 2,2,2-trichloroethyl groups from phosphotriesters of fully protected nucleic acids (e.g., 18a approximately to), having t-butyldimethylsilyl groups at the 2'-positions, leads to the formation of a considerable amount of side-products (e.g., 20 approximately to and 21 approximately to).     

Effect of anaesthetics on calcium-induced luminescence of aequorin

The effect of some general and local anaesthetics on the calcium-induced luminescence of aequorin was studied in vitro using a photomultiplier tube and recording technique. Purified aequorin (0.1 microliter) was injected into a 500 micron diameter porous cellulose acetate capillary tube containing 0.5 M KC1, 20 mM phosphate (pH 7.2) and calcium-EGTA buffers. The trapped aequorin was superfused with buffer solutions which sometimes contained anaesthetic (test) solutions. The results showed that some anaesthetics, e.g. urethane, etomidate and lignocaine, increased whereas others, e.g. methohexitone, thiopentone, decreased the light output (luminescence) of aequorin in constant ionized calcium and EGTA buffers. Similar results were produced by some non-anaesthetic drugs, e.g. glycerol, TEA, caffeine, etc. Concentration-response curves for calcium-dependent and -independent luminescence of aequorin showed that anaesthetics variously affected the aequorin response. Some anaesthetics, e.g. lignocaine, increased the maximum response while others, e.g. etomidate, increased the affinity (i.e. decreased EC50s) of aequorin to calcium ions without altering the slope, which remained at about 2. It was concluded that anaesthetics can either excite or depress aequorin luminescence, the effect being dependent on the type and the concentration used.     

Environmental injury of the lung: role of humoral mediators.

Environmental lung injury may take the form of acute tracheobronchitis, asthma, pulmonary edema, chronic bronchitis, emphysema, allergic pneumonitis, fibrosing alveolitis, pleurisy, and neoplastic disease. Environmental factors eliciting these responses include irritant gases and fumes, oxidants, organic allergens, inorganic dust, bacterial enzymes, and high partial pressures of oxygen. The basic pulmonary reactions to these toxic agents--bronchoconstriction, vasoconstriction, increased vascular permeability, inflammation, carcinogenesis--may be mediated, aggravated, or modulated by biologically active substances. These humoral agents include biogenic amines (e.g. histamine): peptides (e.g., bradykinin, vasoactive intestinal peptide, and spasmogenic lung peptide); enzymes (e.g., proteases, superoxide dismutase, and mixed function oxidases); and acidic lipids (e.g., prostaglandins, prostaglandin endoperoxides, and thromboxanes).     

Nanostructured luminescently labeled nucleic acids

Important and emerging trends at the interface of luminescence, nucleic acids and nanotechnology are: (i) the conventional luminescence labeling of nucleic acid nanostructures (e.g. DNA tetrahedron); (ii) the labeling of bulk nucleic acids (e.g. single‐stranded DNA, double‐stranded DNA) with nanostructured luminescent labels (e.g. copper nanoclusters); and (iii) the labeling of nucleic acid nanostructures (e.g. origami DNA) with nanostructured luminescent labels (e.g. silver nanoclusters). This review surveys recent advances in these three different approaches to the generation of nanostructured luminescently labeled nucleic acids, and includes both direct and indirect labeling methods.     

Functional morphology and mode of life of Palaeozoic leiocope ostracodes

Leiocopes differ from all other major ostracode groups (e.g. Palaeocopa. Binodicopa, Podocopa, Myodocopa) mainly in having a simple rounded/convex shape and a strong (dorsal and/or ventral) asymmetry. These morphological features are highly conservative in leiocopes through the Palaeozoic (lower Ordovician-middle Devonian). Functional morphology and facies distribution suggest that typical 'dome-like' leiocopes (e.g. aparchitids) may have lived off the bottom in relatively deep water environments, contrasting with most (inferred) benthic Palaeozoic ostracodes (e.g. heavily calcified dimorphic palaeocopes). Reversely, smaller sized and both dorsally and ventrally asymmetrical leiocopes (e.g. Ordovician jaanussoniids) are considered as possible benthic dwellers with reduced mobility on or within the substratum. □ Ostracodes, Leiocopa, Aparchitidae, Jaanussoniidae. Palaeozoic. Ordovician, Silurian, Devonian, asymmetry, functional morphology.     

The seasonality of phytoplankton in African lakes

Although some study of the subject began in 1899, wide-ranging information from African water-bodies has only become available since 1950. Important developments included the establishment of long-term centres of research, the adoption of improved methods for quantitative algal sampling, the more intensive study of environmental conditions, the beginnings of experimental testing, and the improvement of taxonomic knowledge.At higher latitudes (> 20 °) examples of pronounced algal seasonality are long-established; they are accompanied and influenced by marked changes in radiant energy income and so water temperature, and often by effects of seasonal water input. Illustrations are given from lakes in Morocco and South Africa.More generally in Africa, including the tropical belt, annual patterns of phytoplankton seasonality are usually either dominated by hydrological features (water input-output) or by hydrographic ones (water-column structure and circulation). Examples of both types are discussed, together with instances (e.g. L. Volta) of combined hydrological and hydrographic regulation. In both the seasonal abundance of diatoms is often distinct and complementary to that of blue-green algae, with differing relationships to vertical mixing and water retention.Horizontal variability in the seasonal cycle is especially pronounced in the larger or morphometrically subdivided lakes. Some inshore-offshore differentiation is also known to affect phytoplankton quantity (e.g. L. George) and species composition (e.g. L. Victoria). Longitudinal differentiation is common in elongate basins especially when with a massive or seasonal inflow at one end (e.g. L. Turkana, L. Nubia, L. Volta); occasional terminal upwelling can also be influential (e.g. southern L. Tanganyika). Such examples grade into the longitudinally differentiated seasonality of flowing river-reservoir systems, as studied on the Blue and White Niles.The annual amplitude of population density, expressed in orders of magnitude (=log₁₀ units), is one measure of seasonal variability. It can exceed 3 orders both in systems subject to hydrological wash-out (e.g. Nile reservoirs) and in the more variable species components of lakes of long retention (e.g. L. Victoria). Low amplitudes can be characteristic of some components (e.g. green algae in L. Victoria) or of total algal biomass (e.g. L. George, L. Sibaya).Seasonal changes may be subordinated to inter-annual ones, especially in shallow and hydrologically unstable lakes (e.g. L. Nakuru).     

Fish impact on rotifer community structure

Larger species, e.g. Asplanchna priodonta and Conochilus unicornis, and grasping species, e.g. Gastropus stylifer and Ascomorpha spp., became more abundant, while smaller filter feeders, e.g. Keratella cochlearis, decreased after an experimental reduction of the fish population. This development is probably caused by changes in basic regulating factors. The change to invertebrate predator dominance may affect smaller species (e.g. Keratella cochlearis) more than others, the increase of net phytoplankton (e.g. peridineans) will favour grasping species, and the change in seasonal occurrence of certain crucial food species may affect the competitive balance between certain rotifers (e.g. Polyarthra spp.).