Spin probe clustering in human erythrocyte ghosts

Summary A model has been developed for 5-nitroxide stearate, I(12,3), distribution in human erythrocyte ghosts which accurately predicts ESR spectral alterations observed with increased probe/total lipid (P/L) at 37°C. This spin probe occupies a class of high-affinity, noninteracting sites at low loading. Saturation occurs with increasing probe concentration, and, at higher loading, the probe inserts itself at initially dilute sites to form membranebound clusters of variable size. No low probe remains at high P/L where all I(12,3) clusters in a concentrated phase. This model allows determination of the dilute/clustered probe ratio, and shows that I(12,3) segregates in erythrocytes at what might otherwise be considered low P/L (e.g., 1/359). These findings validate the earlier use of empirical parameters to estimate probe sequestration in biological membranes.     

Keratinolytic and keratinophilic fungi of children's sandpits in the city of Turin

Soils rich in keratinic residues constitute a permanent or occasional reservoir for dermatophytes and keratinolytic and keratinophilic fungi, and are a source of potential infection for man and animals.The keratinolytic and keratinophilic mycoflora of 28 sandpits in Turin was studied to evaluate the risk of fungal skin infections that is run by children as they play in public parks and to lay the ground-work for a next epidemiological study of soil- and animal-inhabiting dermatophytes found in cities, and their impact on human dermatophytoses. Fifty seven species were isolated and a high percentage (52%) showed keratinolytic activity. Species of Microsporum, Trichophyton, Mariannaea, Aphanoascus, Chrysosporium, Malbranchea and Geomyces showed the most active keratinolysis.Work n. 315 performed under the auspices of the Centro di Studio sulla Micologia del Terreno del C.N.R. at the Dipartimento di Biologia vegetale dell Università di Torino.     

Kinetic characterization of early intermediates in the folding of E. coli tryptophan-synthase beta 2 subunit

This report describes the use of fluorescence energy transfer between an intrinsic energy donor (tryptophan 177) and two chemically added acceptors to study intermediates in the folding of the beta 2 subunit of E. coli tryptophan-synthase. Two early folding steps are thus identified and characterized. One is very rapid (its rate constant at 12 degrees C is 0.02 sec-1) and corresponds to the folding of the N-terminal domain into a structure whose overall features approximate well those of the native domain. The second step is somewhat slower (its rate constant at 12 degrees C is 0.008 sec-1) and involves a conformational rearrangement of the N-terminal domain brought about by the interactions between the N- and C-terminal domains within a monomeric beta chain. This brings to five the number of intermediates which have been identified and ordered on the folding pathway of the dimeric beta 2 subunit.     

Population Dynamics of Meloidogyne incognita and Rotylenchulus reniformis Alone and in Combination,and Their Effects on Sweet Potato

Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) interactions on sweet potato were studied in naturally and artificially infested field plots for 3 years. In a naturally infested field, early season counts of Mi or Rr were positively correlated with later counts of the same nematode, but negative correlations were found between early Mi and subsequent Rr, and early Rr and subsequent Mi counts. In field plots fumigated with methyl bromide and then infested with low levels of Rr, Mi, and Rr + Mi, final population densities of Mi juveniles were reduced by Rr, but Rr was not affected by Mi. In field plots with a high natural population density of Rr, artificial infestation with high levels of Mi in both fumigated and nonfumigated treatments inhibited Rr, while the final Mi juvenile population density was not affected. Results indicate that a competitive interaction exists with each species capable of inhibiting the other and becoming the dominant population. The nematodes had no apparent effect on yield at the inoculum densities used, either alone or mixed. Both nematodes increased cracking of sweet potatoes, but mixed populations did not differ in incidence of cracking from either Rr or Mi alone.     

Energetics of cell-cell and cell-biopolymer interactions

The energy vs distance balance of cell suspensions (in the presence and in the absence of extracellular biopolymer solutions) is studied, not only in the light of the classical Derjaguin-Landau-Verwey-Over-beek (DLVO) theory (which considered just the electrostatic (EL) and Lifshitz-van der Waals (LW) interactions), but also by taking electron-acceptor/electron-donor, or Lewis acid-base (AB) and osmotic (OS) interactions into account. Since cell surfaces, as well as many biopolymers tend to have strong monopolar electron-donor properties, they are able to engage in a strong mutual AB repulsion when immersed in a polar liquid such as water. The effects of that repulsion have been observed earlier in the guise of hydration pressure. The AB repulsion is, at close range, typically one or two orders of magnitude stronger than the EL repulsion, but its rate of decay is much steeper. In most cases, AB interactions are quantitatively the dominant factor in cell stability (when repulsive) and in “hydrophobic interactions” (when attractive). OS interactions exerted by extracellularly dissolved biopolymers are weak, but their rate of decay is very gradual, so OS repulsions engendered by biopolymer solutions may be of importance in certain long-range interactions. OS interactions exerted by biopolymers attached to cells or particles (e.g., by glycocalix glycoproteins), are very short-ranged and usually are negligibly small in comparison with the other interaction forces, in aqueous media.     

Host-plant relationships of lycaenid butterflies: large-scale patterns, interactions with plant chemistry, and mutualism with ants

The Lycaenidae are the second-largest family of butterflies. From host-plant data collated for more than 1200 species worldwide, large-scale taxonomic, geographical and ecological patterns emerge which suggest that phytochemical similarities and barriers, coupled with phylogenetic conservatism and constraints are key factors governing hostplant use. More than two thirds of the lycaenid species are restricted to one plant family or genus. Affiliations with toxic plants are rare in the Lycaenidae, and excretion rather than sequestration of plant toxins appears to be their usual way of detoxifying host-plant compounds. Flavonoids are frequently sequestered by lycaenid larvae and are subsequently concentrated as pigments in the adults' wings, where they might play a role in visual communication. Mutualistic associations with ants occur in the larvae of more than 50% of the extant Lycaenidae species. Because of a conflict between the nutrient demands of the larvae and the proportion of plant-derived resources allocated to maintain the mutualism with ants, variation in resource quality often translates into variation of mutualistic capacities of the caterpillars, in particular under nutrient stress.     

Evidence for an in vivo and in vitro modulation of endogenous cortical GABA release by α-glycerylphosphorylcholine

The effects of α-glycerylphosphorylcholine (α-GPC) on endogenous cortical GABA release were studied both in vivo and in vitro. In freely moving rats, equipped with epidural cups, α-GPC (30–300 mg/kg i.p.) increased GABA release. This effect was potentiated by atropine, both systematically administered (5 mg/kg i.p.) and locally applied (1.4 μM), but not by mecamylamine (4 mg/kg i.p.). The α-GPC-induced increasein GABA release was abolished in rats pretreated with the α₁ receptor antagonist prazosin (14 μg/kg i.p.). In cortical slices α-GPC (0.4 mM) increased the spontaneous GABA efflux. This effectwas abolished by tetrodotoxin (0.5 μM) and prazosin (1 μM), but not by atropine (0.15 μM) ormecamylamine (2.5μM). These results indicate that the facilitatory response by α-GPC on GABArelease does not depend on a direct activation of either muscarinic or nicotinic receptors, but suggest the involvement of the noradrenergic system.     

Site-directed mutagenesis of the CP 47 protein of photosystem II: 167W in the lumenally exposed loop C is required for photosystem II assembly and stability

The intrinsic chlorophyll-protein CP 47 is a component of photosystem II which functions in both light-harvesting and oxygen evolution. Using site-directed mutagenesis we have produced the mutant W167S which lies in loop C of CP 47. This strain exhibited a 75% loss in oxygen evolution activity and grew extremely slowly in the absence of glucose. Examination of normalized oxygen evolution traces indicated that the mutant was susceptible to photoinactivation. Analysis of the variable fluorescence yield indicated that the mutant accumulated very few functional PS II reaction centers. This was confirmed by immunoblotting experiments. Interestingly, when W167S was grown in the presence of 20 M DCMU, the mutant continued to exhibit these defects. These results indicate that tryptophan 167 in loop C of CP 47 is important for the assembly and stability of the PS II reaction center.     

Transmissibility of Bacterial Endosymbionts Between Isolates of Acanthamoeba Spp.

ABSTRACT. Experimental transmission of two bacterial endosymbionts to symbiont-free isolates of Acanthamoeba spp. was studied to determine specificity of the host-symbiont relationship. Both symbionts originated from amoebic isolates displaying an identical mitochondrial DNA Eco RI fingerprint (group AcUW II). Symbioses were readily established in one amoebic isolate which displayed a homologous mtDNA fingerprint (group AcUW II). Exposure of a heterologous amoebic isolate (group AcUW IV) to the two symbionts resulted in either cell death or encystation without the establishment of symbioses. While symbioses were established with an amoebic isolate from a second heterologous group (AcUW I), a unique membranous sheath appeared and persisted around one of the symbionts which did not exist in the original host. An isolate representing a third heterologous amoebic group (AcUW VI) was variable in its susceptibility with one symbiont unable to infect the host and the other becoming established only after an initial reaction in which trophozoites rounded-up and floated off the substrate. These studies suggest that a specific recognition system exists between particular isolates of Acanthamoeba and their symbionts, and that the appearance of a killer phenotype is related to contact between mismatched, though recognized, pairs.     

Contribution of unusual Arginine-Arginine short-range interactions to stabilization and recognition in proteins

Although the majority of the ion pairs found in proteins consists of two charges of opposite sign, the observation of some unusual arrangements of two arginines led us to a search of such occurrences in the Brookhaven Protein Data Bank. We have found 41 Arginine-Arginine interactions with a C...C distance less than 5 å. Computer graphics analysis of these structures shows that most of the Arg-Arg pairs are found in the vicinity of the surface of the proteins, in an easily hydrated region. In order to determine which factors could stabilize such arrangements of species of similar charge, we have carried out AM1 semi-empirical calculations on a model of two guanidinium ions surrounded by several water molecules. The results show the existence of stable clusters with six or more water molecules, with distances between C atoms around 3 å. The bridging role of the water molecules is an important structural and energetic feature and we find bridges of two and three molecules between the guanidinium ions. These results are in good agreement with the structures found in our search of the experimental data. Enhancement of the electrostatic potential around these clusters, when compared to one of the guanidinium ions alone, is also demonstrated.