Disruption of oxidative phosphorylation and synaptic Na,K-ATPase activity by pristanic acid in cerebellum of young rats

Aims

Peroxisomal biogenesis disorders (PBD) are inherited disorders clinically manifested by neurological symptoms and brain abnormalities, in which the cerebellum is usually involved. Biochemically, patients affected by these neurodegenerative diseases accumulate branched-chain fatty acids, including pristanic acid (Prist) in the brain and other tissues.

Main methods

In the present investigation we studied the in vitro influence of Prist, at doses found in PBD, on oxidative phosphorylation, by measuring the activities of the respiratory chain complexes I–IV and ATP production, as well as on creatine kinase and synaptic Na⁺, K⁺-ATPase activities in rat cerebellum.

Key findings

Prist significantly decreased complexes I–III (65%), II (40%) and especially II–III (90%) activities, without altering the activities of complex IV of the respiratory chain and creatine kinase. Furthermore, ATP formation and synaptic Na⁺, K⁺-ATPase activity were markedly inhibited (80–90%) by Prist. We also observed that this fatty acid altered mitochondrial and synaptic membrane fluidity that may have contributed to its inhibitory effects on the activities of the respiratory chain complexes and Na⁺, K⁺-ATPase.

Significance

Considering the importance of oxidative phosphorylation for mitochondrial homeostasis and of Na⁺, K⁺-ATPase for the maintenance of cell membrane potential, the present data indicate that Prist compromises brain bioenergetics and neurotransmission in cerebellum. We postulate that these pathomechanisms may contribute to the cerebellar alterations observed in patients affected by PBD in which Prist is accumulated.     

Seasonal changes of pCO2 over a subantarctic Macrocystis kelp bed

The partial pressure of carbon dioxide (pCO₂), calculated from pH and total alkalinity measurements, was monitored together with chlorophyll a and bacterioplankton biomass in shallow coastal water located inside and outside a giant kelp bed (Macrocystis pyrifera) situated in the Kerguelen Archipelago, Southern Ocean. In spite of large changes over a short time-scale, pCO₂ variations over the year are large and exhibit a seasonal pattern in which the different stages of the annual biological turnover are well marked. The overall pattern of pCO₂ variations is related to biological activity (development of both photosynthesis and respiration) during almost the whole year. However, physical and thermodynamical constraints exert a strong influence on pCO₂ at meso time-scale (10 days) and/or when biological activity is weak. Macrocystis acts to maintain pCO₂ below saturation almost the whole year and large undersaturations (pCO₂ as low as 20 μatm) were observed within the kelp bed. Furthermore, primary production of Macrocystis covers a period of 8 ∼ 9 months a year from winter to late summer and the kelp bed seems to favour the spring phytoplanktonic bloom. The buffer factor β indicates that, outside the kelp bed, inorganic carbon dynamics are mainly influenced by air-sea exchange and photosynthesis without calcification. Inside the kelp bed, β suggests calcification by the epiphytic community. Accepted: 1 April 2000     

Assessment of river condition at a large spatial scale using predictive models

1. RIVPACS-type predictive models were developed at a relatively large spatial scale for the Australian state called New South Wales (NSW, 801 428 km²). Aquatic macroinvertebrate samples and physical and chemical data were collected from 250 reference sites (little affected by human activities) and 23 test sites (with known human impacts) throughout NSW in autumn and spring 1995 and identified mostly to family level. Reference sites were grouped based on their macroinvertebrate data using classification (UPGMA) and ordination techniques. Relationships between macroinvertebrate and environmental data were established using principal axis correlations and stepwise multiple discriminant function analysis. models for predicting invertebrate assemblages were developed separately for edge and riffle habitats for autumn and spring data sets and for combined autumn and spring data sets. 2. Sites in the lowland sections of the western flowing rivers were characterized by low taxonomic richness and were distinct from the sites in the eastern part of the state. Sites on the western slopes of the Great Dividing Range in southern and northern NSW mostly fell into separate groups. In eastern NSW, site groups did not represent a north, central and south division. Sites on highland streams, coastal fringe streams and large rivers mostly formed distinct groups, but most of the sites on east-flowing rivers fell into large site groups that did not have clear geographic boundaries. 3. Environmental variables that were strongly correlated with ordinations of macroinvertebrate presence/absence at sites were water temperature, altitude, longitude and maximum distance from source. The predictor variables determined by DFA for the six models created included alkalinity, altitude, location (longitude and/or latitude), stream size and substratum composition. These are generally in common with the variables determined for other large geographic areas in Australia and the United Kingdom. 4. Model outputs from reference sites suggest that, among the six models, the riffle model combining autumn and spring is likely to give the most reliable predictions. The combined edge model also performed well but refinements are needed for single season models to provide reliable outputs. 5. Combined season models both for riffles and for edges detected biological impairment at all but one of the test sites. Single season riffle models also detected impairment while single season edge models characterized sites as unimpaired despite other models’ indications of impaired fauna. Riffle models may be more sensitive than edge models but the sampling of riffles is often limited by flow. Edge habitats are available at most sites but there may be few riffles in floodplain rivers. Available resources, desired model sensitivity, and river type should be considered jointly to determine the most useful habitat to sample.     

Structure of the mammalian antimicrobial peptide Bac7(1–16) bound within the exit tunnel of a bacterial ribosome

Proline-rich antimicrobial peptides (PrAMPs) produced as part of the innate immune response of animals, insects and plants represent a vast, untapped resource for the treatment of multidrug-resistant bacterial infections. PrAMPs such as oncocin or bactenecin-7 (Bac7) interact with the bacterial ribosome to inhibit translation, but their supposed specificity as inhibitors of bacterial rather than mammalian protein synthesis remains unclear, despite being key to developing drugs with low toxicity. Here, we present crystal structures of the Thermus thermophilus 70S ribosome in complex with the first 16 residues of mammalian Bac7, as well as the insect-derived PrAMPs metalnikowin I and pyrrhocoricin. The structures reveal that the mammalian Bac7 interacts with a similar region of the ribosome as insect-derived PrAMPs. Consistently, Bac7 and the oncocin derivative Onc112 compete effectively with antibiotics, such as erythromycin, which target the ribosomal exit tunnel. Moreover, we demonstrate that Bac7 allows initiation complex formation but prevents entry into the elongation phase of translation, and show that it inhibits translation on both mammalian and bacterial ribosomes, explaining why this peptide needs to be stored as an inactive pro-peptide. These findings highlight the need to consider the specificity of PrAMP derivatives for the bacterial ribosome in future drug development efforts.     

Structure of a human pre-40S particle points to a role for RACK1 in the final steps of 18S rRNA processing

Synthesis of ribosomal subunits in eukaryotes is a complex and tightly regulated process that has been mostly characterized in yeast. The discovery of a growing number of diseases linked to defects in ribosome biogenesis calls for a deeper understanding of these mechanisms and of the specificities of human ribosome maturation. We present the 19 Å resolution cryo-EM reconstruction of a cytoplasmic precursor to the human small ribosomal subunit, purified by using the tagged ribosome biogenesis factor LTV1 as bait. Compared to yeast pre-40S particles, this first three-dimensional structure of a human 40S subunit precursor shows noticeable differences with respect to the position of ribosome biogenesis factors and uncovers the early deposition of the ribosomal protein RACK1 during subunit maturation. Consistently, RACK1 is required for efficient processing of the 18S rRNA 3′-end, which might be related to its role in translation initiation. This first structural analysis of a human pre-ribosomal particle sets the grounds for high-resolution studies of conformational transitions accompanying ribosomal subunit maturation.     

European Lampreys: New Insights on Postglacial Colonization,Gene Flow and Speciation

Ice ages are known to be the most dominant palaeoclimatic feature occurring on Earth, producing severe climatic oscillations and consequently shaping the distribution and the population structure of several species. Lampreys constitute excellent models to study the colonization of freshwater systems, as they commonly appear in pairs of closely related species of anadromous versus freshwater resident adults, thus having the ability to colonize new habitats, through the anadromous species, and establish freshwater resident derivates. We used 10 microsatellite loci to investigate the spatial structure, patterns of gene flow and migration routes of Lampetra populations in Europe. We sampled 11 populations including the migratory L. fluviatilis and four resident species, L. planeri, L. alavariensis, L. auremensis and L. lusitanica, the last three endemic to the Iberian Peninsula. In this southern glacial refugium almost all sampled populations represent a distinct genetic cluster, showing high levels of allopatric differentiation, reflecting long periods of isolation. As result of their more recent common ancestor, populations from northern Europe are less divergent among them, they are represented by fewer genetic clusters, and there is evidence of strong recent gene flow among populations. These previously glaciated areas from northern Europe may have been colonized from lampreys expanding out of the Iberian refugia. The pair L. fluviatilis/L. planeri is apparently at different stages of speciation in different locations, showing evidences of high reproductive isolation in the southern refugium, and low differentiation in the north.     

Action de molecules a proprietes hormonales sur l'activite phenylalanine ammoniac lyase

A comparative study has been made of phenylalanine ammonia lyase (PAL) activity in plants sensitive or resistant to various herbicides (piclorame, methylchloro-phenoxyacetic acid (MCPA), atrazine). Piclorame, a herbicide with hormonal activity caused a large decrease in PAL activity of sensitive plants (Nicotiana tabacum), even at low concentrations (5 × 10^-9M) whilst in resistant plants (Triticum aestivum) its effect is negligible; MCPA, also a herbicide with hormonal activity, similarly affects the activity of PAL, but only at higher concentrations. On the contrary, the action of atrazine, which has no hormonal activity, is lower and weaker, probably being only a secondary effect. Determinations of PAL activity during the photoperiod following piclorame application indicate that this herbicide influences principally the photodependent enzyme activity.