Myosin motors and not actin comets are mediators of the actin-based Golgi-to-endoplasmic reticulum protein transport

We have previously reported that actin filaments are involved in protein transport from the Golgi complex to the endoplasmic reticulum. Herein, we examined whether myosin motors or actin comets mediate this transport. To address this issue we have used, on one hand, a combination of specific inhibitors such as 2,3-butanedione monoxime (BDM) and 1-[5-isoquinoline sulfonyl]-2-methyl piperazine (ML7), which inhibit myosin and the phosphorylation of myosin II by the myosin light chain kinase, respectively; and a mutant of the nonmuscle myosin II regulatory light chain, which cannot be phosphorylated (MRLC2(AA)). On the other hand, actin comet tails were induced by the overexpression of phosphatidylinositol phosphate 5-kinase. Cells treated with BDM/ML7 or those that express the MRLC2(AA) mutant revealed a significant reduction in the brefeldin A (BFA)-induced fusion of Golgi enzymes with the endoplasmic reticulum (ER). This delay was not caused by an alteration in the formation of the BFA-induced tubules from the Golgi complex. In addition, the Shiga toxin fragment B transport from the Golgi complex to the ER was also altered. This impairment in the retrograde protein transport was not due to depletion of intracellular calcium stores or to the activation of Rho kinase. Neither the reassembly of the Golgi complex after BFA removal nor VSV-G transport from ER to the Golgi was altered in cells treated with BDM/ML7 or expressing MRLC2(AA). Finally, transport carriers containing Shiga toxin did not move into the cytosol at the tips of comet tails of polymerizing actin. Collectively, the results indicate that 1) myosin motors move to transport carriers from the Golgi complex to the ER along actin filaments; 2) nonmuscle myosin II mediates in this process; and 3) actin comets are not involved in retrograde transport.     

Influence of larval rearing diet on the intensive searching behaviour ofHarmonia axyridis [Col.: Coccinellidae] Larvae

The capture of prey by last instar larvae of the aphidophagous coccinellidHarmonia axyridis Pallas (Col.,Coccinellidae) modified larval movement so that extensive search was replaced by intensive search. The continuous rearing of this species on eggs ofEphestia kühniella Zell. (Lep.,Pyralidae) or on the aphidAcyrthosiphon pisum Harris (Homopt.,Aphidae) led to conditioning. Only larvae that ate the same prey as they were reared on, adopted intensive searching movements after feeding. Using larvae reared on substitute prey in biological control systems may decrease the efficiency of the released predators.     

Determination of the aza alkyl lysophospholipid 3-methoxy-2-N,N-methyloctadecylaminopropyloxyphosphorylcholine in rat plasma by liquid chromatography—particle beam—mass spectrometry

A sensitive and specific method for the determination of the aza alkyl lysophospholipid (AALP) 3-methoxy-2-N,N-methyloctadecylaminopropyloxyphosphorylcholine (I) in rat plasma is described. The target molecule was analyzed by high-performance liquid chromatography (HPLC)—mass spectrometry (MS) after one single liquid—liquid extraction with chloroform—methanol (2:1, v/v). 1,2-Didecanoyl-sn-glycero-3-phosphocholine was used as internal standard. HPLC was carried out using a polymeric reversed-phase column; the coupling to the mass spectrometer was a particle beam (PB) interface, and the ionization method was electron impact (EI). This simple and rugged method permits the measurement of I in rat plasma in the range of 25 ng/ml–5 μg/ml with good accuracy and precision and is used in pharmacokinetic studies.     

Locomotor Behavior of Flightless Harmonia axyridis Pallas (Col., Coccinellidae)

To improve the efficiency of the lady beetle H. axyridis as a biological control agent against aphids, a flightless population was obtained by feeding adults with a mutagen and selecting their progeny for nonflying but otherwise morphologically normal individuals. These flightless adults attempted to fly but immediately fell. They softened their fall by opening their elytrae and wings. The inability to fly could result from change in their flying behavior compared to control adults. The flight duration was very much shorter, and the wing beat frequency and, more particularly, the amplitude of the wing beats were clearly lower. More time was spent in the other components of the flight behavior such as wing rotation, wing immobility, and wing folding. The sequence of these patterns differed slightly, due mainly to change in their frequency. The locomotor behavior was not modified by the mutation, which affected only the wing muscles. Searching behavior of mutant adults differed from that of control adults only in that they took longer to encounter and ingest aphids. Nevertheless, the larval growth and reproductive rate remained unchanged. The behavioral and biological features of these flightless adults indicate that it should be possible to use them in biological control programs.     

Effects of fire recurrence in Quercus coccifera L. shrublands of the Valencia Region (Spain): II. plant and soil nutrients

The consequences of fire recurrence (1, 2 and 3 fires in 16 years) on plant and soil C, N, P and K from Quercus coccifera garrigues were analysed in the Valencia Region (E Spain). Plant and forest floor (L horizon) nutrient concentrations either changed weakly or showed no change with fire recurrence. At the soil surface (0-2.5 cm), soil potential mineralisable nitrogen increased and available P decreased after an initial increase, whereas exchangeable K was not affected by successive fires. However, the significance of those observed trends for N and P was site-dependent. Despite the rapid formation of the L soil horizon, fire recurrence did not permit the development of the whole forest floor profile observed in the unburned garrigues. These organic layers contained a great proportion of the total nutrient pool, especially for N. Forest-floor and aboveground plant combustion by fire may produce significant losses of N and P compared with those available in mineral soil, whereas soil exchangeable K is large enough to replace these losses. Belowground nutrient reserves may account for the quick recovery of the Quercus coccifera aboveground biomass although successive fires could deplete these reserves and produce a loss of biomass and productivity in this species.     

Relevance of ammonium oxidation within biological soil crust communities

Thin, vertically structured topsoil communities that become ecologically important in arid regions (biological soil crusts or BSCs) are responsible for much of the nitrogen inputs into pristine arid lands. We studied N(2) fixation and ammonium oxidation (AO) at subcentimetre resolution within BSCs from the Colorado Plateau. Pools of dissolved porewater nitrate/nitrite, ammonium and organic nitrogen in wetted BSCs were high in comparison with those typical of aridosoils. They remained stable during incubations, indicating that input and output processes were of similar magnitude. Areal N(2) fixation rates (6.5-48 micromol C(2)H(2) m(-2) h(-1)) were high, the vertical distribution of N(2) fixation peaking close to the surface if populations of heterocystous cyanobacteria were present, but in the subsurface if they were absent. Areal AO rates (19-46 micromol N m(-2) h(-1)) were commensurate with N(2) fixation inputs. When considering oxygen availability, AO activity invariably peaked 2-3 mm deep and was limited by oxygen (not ammonium) supply. Most probable number (MPN)-enumerated ammonia-oxidizing bacteria (6.7-7.9 x 10(3) cells g(-1) on average) clearly peaked at 2-3 mm depth. Thus, AO (hence nitrification) is a spatially restricted but important process in the nitrogen cycling of BSC, turning much of the biologically fixed nitrogen into oxidized forms, the fate of which remains to be determined.     

Microbial diversity of benthic mats along a tidal desiccation gradient

We investigated the influence of desiccation frequency, indicated by tidal position, on microbial community structure, diversity and richness of microbial mats. We independently characterized cyanobacterial, bacterial and archaeal communities, and their spatial variability for two distinct microbial mat systems: subtidal hypersaline mats and intertidal sand flat mats. Community fingerprints based on 16S rDNA were obtained via denaturing gradient gel electrophoresis using polymerase chain reaction primers specific for each group. Fingerprints for all three groups were consistently similar [> or =85% according to Weighted Pair Group with Arithmetic Mean (WPGMA) analysis] along a 1-km-long transect in subtidal mats. Here, pair-wise comparison analysis yielded minimal variation in diversity and richness for all groups. Fingerprints of three sites along an intertidal transect were heterogenous (> or =32% similarity according to WPGMA analysis) with clear shifts in community structure in all three microbial groups. Here, all groups exhibited statistically significant decreases in richness and diversity with tidal height (as desiccation frequency increases). Regression analysis yielded a strong correlation between diversity or richness estimates and position along the tidal gradient, for both Archaea and Bacteria, with Cyanobacteria exhibiting a weaker correlation. These results suggest that desiccation frequency can shape the structure of microbial mat communities, with Archea being least tolerant and Cyanobacteria most tolerant.