Experimental evidence of the dynamic effect of free-floating plants on phytoplankton ecology

1. There is an increasing concern to understand the role of free-floating plants (FFP) on the functioning and structure of shallow lakes, particularly the mechanisms by which their dominance is self-stabilizing and how they may outcompete phytoplankton.
2. In a field experiment with mesocosms, we simulated three commonly encountered scenarios in warm temperate shallow lakes: FFP dominance, FFP fluctuation and FFP absence. We explored the effects of several key processes, triggered by FFP dynamics, on the composition, diversity and production of phytoplankton, and on the physicochemical conditions.
3. The effects of persistent floating mats on light interception triggered a complex response by the phytoplankton: species were lost and biomass was low, yet high diversity and productivity potential were maintained. A contrasting phytoplankton response characterized mesocosms lacking FFP, where light was sufficient but nitrogen was limiting. Fluctuating FFP cover brought periodic shifts between these two limiting resources for the phytoplankton, which most probably explain the maintenance of richness, diversity and production in these shallow lakes.
4. These results support the recently proposed framework of alternative stable states, providing experimental field evidence of the mechanisms resulting from the shifts between floating plants and phytoplankton dominance.     

Physical aspects of dynamic stereotactic radiosurgery

Dynamic stereotactic radiosurgery is a radiosurgical technique based on a medium-energy isocentric linear accelerator and a stereotactic frame. The technique uses concurrent and continuous rotations of both the gantry (300 degrees, from 30 to 330 degrees) and the couch (150 degrees, from 75 to -75 degrees). It gives a uniform dose (+/- 5%) within the target volume and dose fall-offs outside the target volume comparable to those obtained from presently known radiosurgical techniques.     

Experimental evidence for lack of homodimerization of the G protein-coupled human N-formyl peptide receptor

A large number of G protein-coupled receptors have been shown to form homodimers based on a number of different techniques such as receptor coimmunoprecipitation, cross-linking, and fluorescence resonance energy transfer. In addition, functional assays of cells coexpressing a mutant receptor with a wild-type receptor have shown receptor phenotypes that can best be explained through dimerization. We asked whether the human neutrophil N-formyl peptide receptor (FPR) forms dimers in Chinese hamster ovary cells by coexpressing wild-type FPR with one of two mutants: D71A, which is uncoupled from G protein, and N297A, which has a defect in receptor phosphorylation and endocytosis. Experiments measuring chemotaxis, ligand-induced release of intracellular calcium, and p42/44 mitogen-activated protein kinase activation did not show an inhibitory effect of the coexpressed FPR D71A mutant. Coexpressed wild-type receptor was efficiently internalized, but failed to correct the endocytosis defects of the D71A and the N297A mutants. To explore the possibility that the mutations themselves prevented dimerization, we examined the coimmunoprecipitation of differentially epitope-tagged FPR. Immunoprecipitation of hemagglutinin-tagged FPR failed to coimmunoprecipitate coexpressed c-myc-tagged FPR and vice versa. Together, these data suggest that, unlike many other G protein-coupled receptors, FPR does not form homodimers.     

Evolutionary aspects of gonadotropin-releasing hormone and its receptor

Summary 1. Gonadotropin-releasing hormone (GnRH) was originally isolated as a hypothalamic peptide hormone that regulates the reproductive system by stimulating the release of gonadotropins from the anterior pituitary. However, during evolution the peptide was subject to gene duplication and structural changes, and multiple molecular forms have evolved.2. Eight variants of GnRH are known, and at least two different forms are expressed in species from all vertebrate classes: chicken GnRH II and a second, unique, GnRH isoform.3. The peptide has been recruited during evolution for diverse regulatory functions: as a neurotransmitter in the central and sympathetic nervous systems, as a paracrine regulator in the gonads and placenta, and as an autocrine regulator in tumor cells.4. Evidence suggests that in most species the early-evolved and highly conserved chicken GnRH II has a neurotransmitter function, while the second form, which varies across classes, has a physiologic role in regulating gonadotropin release.5. We review here evolutionary aspects of the family of GnRH peptides and their receptors.     

Structural dynamic aspects of protein synthesis on ribosomes

Three types of conformational changes in the translating ribosome are considered: (1) intersubunit movement (ribosome unlocking) during translocation; (2) L7/L12 stalk mobility affected by elongation factors; (3) change of tRNA residue during its transition from the A-site to the P-site. Relevant experimental data are reviewed.     

Phosphorylation of brain muscarinic receptor: evidence of receptor regulation

Muscarinic receptor, from porcine synaptic membrane, was purified by affinity chromatography. Molecular weight analysis by SDS-gel electrophoresis revealed one major peptide with an apparent Mr of 68 +/- 2 Kda. The purified receptor was phosphorylated by the catalytic subunit of cAMP-dependent protein kinase resulting in a concomitant loss in specific binding, and this loss was reversed by calcineurin.     

Impulse frequency encoding of the dynamic aspects of excitation

Like receptors, neurones of the mammalian CNS react to both the static and the dynamic features of the input signals. The quantitative aspect of the current-to-frequency transduction during input transients were analyzed in spinal motoneurones and in corticospinal neurones by the technique of injecting intracellularly either ramp of sine-wave currents. It was found that, in both types of neurones, the instantaneous firing frequency is linearly correlated, within definite ranges, both to the intensity and to the velocity of change of the input currents. By recording the isometric mechanogram of the muscle units innervated by the impaled motoneurones, it was disclosed that the motoneurones discharges produced by the ramp currents develop tension changes, whose average velocity is proportional to the ramp slope. For both types of neurones, results are consistent with the hypothesis that the major determinants of the double sensitivity, to the intensity and to the velocity, are the kinetics of the potassium conductance system responsible for the spike afterhyperpolarization.     

Experimental aspects of in vitro and in vivo photochemotherapy

The selectivity of in vitro photodynamic reactions and the in vivo effects induced by PRT, whether the irradiation is applied interstitially or externally, still remains unclear. In vitro studies were performed using leukemic cell lines and syngeneic normal hemopoietic progenitors. For these, cells incubated with hematoporphyrin derivative (HPD) and non-incubated cells were irradiated with an argon laser. Data were obtained as the count of cell colonies found after a 7-day incubation period on semi-solid collagen gel medium. In vivo studies employed the HT 29 tumor model grafted into nude mice. Both animals injected with HPD and non-infected controls were irradiated with a dye laser pumped by an argon laser (Coherent) using a 400 micron optic fiber located either at a distance of 65 mm from the skin or inserted into the tumor. The temperature increase occurring during PRT was measured using non-absorbing thermocouples. In vitro, after HPD treatment and argon irradiation leukemic cells showed a greater phototoxicity (greater than 2 log10) than did the normal cells (0.25 log10). In vivo, when the heat rise is very similar (less than 4 degrees C) in both the tissues irradiated externally and those irradiated interstitially after HPD injection, histological examination of these did not reveal any quantitative differences (90% of tumor mass). These results are discussed.     

Experimental evidence of dynamic self-organization in the electron transfer system (example of reaction centers of purple bacteria)

The results of an experimental study of nonlinear dynamic processes in the electron transfer system, the reaction centers (RCs) of purple bacteria are presented. A difference was observed in the absorption spectra of RCs exposed to a rising intensity of acting light compared to a descending intensity of acting light. We observed the hysteresis of the RC optical transmission coefficient at =865 nm, with a quasistationary increase and subsequent decrease of the optical excitation level. The kinetics of charge recombination in an RC containing two quinone acceptors revealed a dependence on the prehistory of the RC illumination. The results were interpreted in terms of the existence of a light-induced memory effect in the electron-conformational system and the appearance of bifurcation in the system at critical values of the photoinduced electron flux through the macromolecule.     

Lactoferrin and Iron: structural and dynamic aspects of binding and release

Lactoferrin (Lf) has long been recognized as a member of the transferrin family of proteins and an important regulator of the levels of free iron in the body fluids of mammals. Its ability to bind ferric iron with high affinity (KD approximately 10(-20) M) and to retain it to low pH gives the protein bacteriostatic and antioxidant properties. This ability can be well understood in terms of its three dimensional (3D) structure. The molecule is folded into two homologous lobes (N- and C-lobes) with each lobe binding a single Fe3+ ion in a deep cleft between two domains. The iron sites are highly conserved, and highly favorable for iron binding. Iron binding and release are associated with large conformational changes in which the protein adopts either open or closed states. Comparison of available apolactoferrin structures suggests that iron binding is dependent on the dynamics of the open state. What triggers release of the tightly bound iron, however, and why lactoferrin retains iron to much lower pH than its serum homologue, transferrin, has been the subject of much speculation. Comparisons of structural and functional data on lactoferrins and transferrins now suggest that the key factor comes from cooperative interactions between the two lobes of the molecule, mediated by two alpha-helices.     

Naming dynamic and static actions: neuropsychological evidence.

There has been considerable interest in identifying the neural correlates of action naming, but the bulk of previous work on this topic has utilized static stimuli. Recent research comparing the visual processing of dynamic versus static actions suggests that these two types of stimuli engage largely overlapping neural systems, raising the possibility that the higher-order processing requirements for naming dynamic and static actions might not be very different. To explore this issue in greater depth, we developed the Dynamic Action Naming Test (DANT), which consists of 158 video clips 3-5s in length, for each of which the participant is asked to produce the most appropriate verb. We administered the DANT to 78 brain-damaged patients drawn from our Patient Registry, and to a demographically matched group of 50 normal participants. Out of the 16 patients who performed defectively on the DANT, nearly all (15/16) had damage in the left hemisphere. Lesion analysis indicated that the frontal operculum was the most frequent area of damage in the 15 patients; also, damage to the posterolateral temporal-occipital sector (in and near MT) was specifically related to defective dynamic action naming. Most of the brain-damaged participants (n=71) also received our Static Action Naming Test (SANT), and we found that performances on verb items that were common across the DANT and SANT were highly correlated (R=.91). Moreover, patients who failed the DANT almost invariably also failed the SANT. These findings lend further support to the hypothesis that there is considerable commonality in the neural systems underlying the use of verbs to orally name dynamic and static actions, a conclusion that is in turn compatible with the concept of "representational momentum". Our results also contribute more generally to the rapidly growing field of research on embodied cognition.