Molecular genetics of cell death in the nematode Caenorhabditis elegans

In C. elegans, cell death can be readily studied at the cellular, genetic, and molecular levels. Two types of death have been characterized in this nematode: (1) programmed cell death, which occurs as a normal component in development; and (2) pathological cell death which occurs aberrantly as a consequence of mutation. Analysis of mutations that disrupt programmed cell death in various ways has defined a genetic pathway for programmed cell death which includes genes that perform such functions as the determination of which cells die, the execution of cell death, the engulfment of cell corpses, and the digestion of DNA from dead cells. Molecular analysis is providing insightinto the nature of the molecules that function in these aspects of programmed cell death. Characterization of some genes that mutate to induce abnormal cell death has defined a novel gene family called degenerins that encode putative membrane proteins. Dominant alleles of at least two degenerin genes, mec-4 and deg-1, can cause cellular swelling and late onset neurodegeneration of specific groups of cells. © 1992 John Wiley & Sons, Inc.     

Getting a grip on the intertidal: flow microhabitat and substratum type determine the dislodgement of the crab Pachygrapsus crassipes (Randall) on rocky shores and in estuaries

Hydrodynamic forces can affect survival as well as limit the movement of motile benthic animals. An animal's danger of dislodgement depends on the hydrodynamic forces it experiences in its microhabitat relative to the force required to dislodge it (tenacity) from the substratum. We measured water flow and substratum characteristics in two different habitats of the shore crab Pachygrapsus crassipes: a wave-swept rocky shore and an intertidal mudflat. The maximum water velocities and accelerations in the microhabitats of the crabs at the wave-swept site were three times and two times greater, respectively, than at the mudflat site. In the laboratory, we measured the tenacity of crabs of various sizes on different substrata, and also measured their drag, lift and added-mass coefficients. Using these data, we calculated the flow conditions under which crabs would be overturned or sheared off the substratum in their two habitats. The net horizontal force (drag plus acceleration reaction) required to dislodge a crab on a rugose rock substratum was an order of magnitude greater than on smooth rock and two orders of magnitude greater than on mud. Our calculations indicate that, under non-storm conditions, crabs will not be dislodged from the substratum in either the mudflat or the wave-swept habitat when grasping the substratum with maximum tenacity. Moving crabs have lower tenacity and our calculations predict that hydrodynamic forces will restrict the mobility of large crabs more than that of small ones on smooth, but not on rugose rock.     

Dimer–monomer equilibrium of human thymidylate synthase monitored by fluorescence resonance energy transfer

An ad hoc bioconjugation/fluorescence resonance energy transfer (FRET) assay has been designed to spectroscopically monitor the quaternary state of human thymidylate synthase dimeric protein. The approach enables the chemoselective engineering of allosteric residues while preserving the native protein functions through reversible masking of residues within the catalytic site, and is therefore suitable for activity/oligomerization dual assay screenings. It is applied to tag the two subunits of human thymidylate synthase at cysteines 43 and 43′ with an excitation energy donor/acceptor pair. The dimer–monomer equilibrium of the enzyme is then characterized through steady‐state fluorescence determination of the intersubunit resonance energy transfer efficiency.     

Collagen targeting using multivalent protein-functionalized dendrimers

Collagen is an attractive marker for tissue remodeling in a variety of common disease processes. Here we report the preparation of protein dendrimers as multivalent collagen targeting ligands by native chemical ligation of the collagen binding protein CNA35 to cysteine-functionalized dendritic divalent (AB₂) and tetravalent (AB₄) wedges. The binding of these multivalent protein constructs was studied on collagen-immobilized chip surfaces as well as to native collagen in rat intestinal tissues. To understand the importance of target density we also created collagen-mimicking surfaces by immobilizing synthetic collagen triple helical peptides at various densities on a chip surface. Multivalent display of a weak-binding variant (CNA35-Y175K) resulted in a large increase in collagen affinity, effectively restoring the collagen imaging capacities for the AB₄ system. In addition, dissociation of these multivalent CNA35 dendrimers from collagen surfaces was found to be strongly attenuated.     

Cluster analysis of behavioural and event-related potentials during a contingent negative variation paradigm in remitting-relapsing and benign forms of multiple sclerosis

Background  

Event-related potentials (ERPs) may be used as a highly sensitive way of detecting subtle degrees of cognitive dysfunction. On the other hand, impairment of cognitive skills is increasingly recognised as a hallmark of patients suffering from multiple sclerosis (MS). We sought to determine the psychophysiological pattern of information processing among MS patients with the relapsing-remitting form of the disease and low physical disability considered as two subtypes: 'typical relapsing-remitting' (RRMS) and 'benign MS' (BMS). Furthermore, we subjected our data to a cluster analysis to determine whether MS patients and healthy controls could be differentiated in terms of their psychophysiological profile.     

Multiphasic On/Off Pheromone Signalling in Moths as Neural Correlates of a Search Strategy

Insects and robots searching for odour sources in turbulent plumes face the same problem: the random nature of mixing causes fluctuations and intermittency in perception. Pheromone-tracking male moths appear to deal with discontinuous flows of information by surging upwind, upon sensing a pheromone patch, and casting crosswind, upon losing the plume. Using a combination of neurophysiological recordings, computational modelling and experiments with a cyborg, we propose a neuronal mechanism that promotes a behavioural switch between surge and casting. We show how multiphasic On/Off pheromone-sensitive neurons may guide action selection based on signalling presence or loss of the pheromone. A Hodgkin-Huxley-type neuron model with a small-conductance calcium-activated potassium (SK) channel reproduces physiological On/Off responses. Using this model as a command neuron and the antennae of tethered moths as pheromone sensors, we demonstrate the efficiency of multiphasic patterning in driving a robotic searcher toward the source. Taken together, our results suggest that multiphasic On/Off responses may mediate olfactory navigation and that SK channels may account for these responses.     

The use of GMOs (genetically modified organisms): agricultural biotechnology or agricultural biopolitics?

Agricultural biotechnologies embrace a large array of conventional and modern technologies, spanning from composting organic by-products of agriculture to innovative improvement of quality traits of about twenty out of the mostly cultivated plants. In EU a rather restrictive legislative framework has been installed for GMOs, requiring a risk assessment disproportionate with respect to conventional agriculture and organic farming products. The latter are far from being proved safe for human and animal health, and for the environment.Biotechnology of GMOs has been overtaken by biopolitics. On one side there are biotechnological challenges to be tackled, on another side there is plenty of ground for biopolitical decisions about GMOs. Perhaps the era of harsh confrontation could be fruitfully replaced by sensible cooperation, in order to get a sustainable agricultural development.     

Determining [H]Thymidine Incorporation into Bacterioplankton DNA: Improvement of the Method by DNase Treatment

Determination of [H] thymidine incorporation into bacterial DNA versus other macromolecules is usually achieved by NaOH and hot trichloroacetic acid hydrolysis. This procedure was found not to be specific enough. An alternative method founded on DNase treatment is proposed. Under the new method, the fraction of thymidine incorporated into DNA ranged from 10 to 83%.