Why preferential hydration does not always stabilize the native structure of globular proteins

The observed preferential hydration of proteins in aqueous MgCl2 solutions at low pH and low salt concentration (Arakawa et al., 1990) prompted a scrutiny of possible protein stabilization by MgCl2 under the same conditions, in view of earlier observations in aqueous solutions of sugars, amino acids, and a number of salts that preferential hydration is usually accompanied by the stabilization of the native structure of globular proteins. The results of thermal transition experiments on five proteins (ribonuclease A, lysozyme, beta-lactoglobulin, chymotrypsinogen, and bovine serum albumin) revealed neither significant stabilization nor destabilization of the protein structures by MgCl2 both at acid conditions (except for ribonuclease A, which was stabilized, but to a much smaller extent than by MgSO4) and at higher pH at which MgCl2 displayed little preferential hydration. This was in contrast to the great stabilizing action of MgSO4 at the same conditions. 2-Methyl-2,4-pentanediol (MPD), which gives a very large preferential hydration of native ribonuclease A at pH 5.8 [Pittz & Timasheff (1978) Biochemistry 17, 615-623], was found to be a strong destabilizer of that protein at the same conditions. Analysis of the preferentially hydrating solvent systems led to their classification into two categories: those in which the preferential hydration is independent of solution conditions and those in which it varies with conditions. The first always stabilize protein structure, while the second do not. In the first category the predominant interaction is that of cosolvent exclusion, determined by solvent properties, with the protein being essentially inert. In the second category interactions are determined to a major extent by the chemical nature of the protein surface.(ABSTRACT TRUNCATED AT 250 WORDS)     

Strength, drag, and dislodgment of two competing intertidal algae from two wave exposures and four seasons

Intertidal macroalgae often experience greater risk of dislodgment with increasing size because of underscaling of breaking force of their stipes relative to drag on their thalli. This ratio (breaking force/drag) indicates safety from breakage at a given flow speed, with values greater than one indicating safety from breakage and values lower than one indicating danger of breakage. We examined this force ratio for the largest thalli of two species of co-dominant, red algae, Chondrus crispus Stackhouse and Mastocarpus stellatus Stack. In With. (Guiry), in four seasons at two wave exposures. During fall and winter, the largest thalli in both populations were dislodged resulting in a decrease in mass of the largest thalli found. This decrease was greater for Chondrus than for Mastocarpus, but their mass-specific force ratios (at 0.55 m s⁻¹) were equal indicating similar size-specific risk of dislodgment. The equality of force ratios was underlain by two similarities: (1) breaking force was independent of mass and not different between species; (2) mass-specific drag was not different between species. These similarities were underlain by dissimilar causes: (i) similarity in breaking force (the product of cross-sectional area and material strength) occurred because greater material strength of Mastocarpus compensated for greater mass-specific cross-sectional area of Chondrus; (ii) similarity in mass-specific drag (a function of planform area and the coefficient of drag) occurred because greater drag coefficients for Mastocarpus compensated for greater mass-specific planform areas of Chondrus. The similarity in force ratio, if it held at season- and site-relevant flow speeds, would suggest that during seasons of minimal growth and high wave exposure, the mass of the largest thalli of both species should be the same. Chondrus, however, had a greater mass at both sites in all seasons. Chondrus experienced greater decreases in mass probably because it grew larger and larger thalli are less safe. Extrapolation of a site-relevant force ratio for Chondrus in the fall revealed (1) that the site-relevant force ratio did not differ between exposures even though the mass-specific force ratio was greater at the protected site, and (2) a paradoxical result that all Chondrus thalli studied ought to have dislodged, but had not. This paradox may be resolved by consideration of the protection conferred by canopies of Chondrus: a canopy may effectively raise its site-relevant force ratio. Perhaps differences in protection conferred by different canopies explain why larger Chondrus persist with Mastocarpus even given a similarity in mass-specific force ratio.     

Response of deposit-feeders to exclusion of epibenthic predators in a Mediterranean intertidal flat

Deposit-feeders are common components of macrofaunal assemblages in intertidal soft sediments. Predation has been considered to have a central role in affecting their distribution and population dynamics. This study investigates the effect of epibenthic predators on deposit-feeders, inhabiting the shallow layers of the sediment (surface and subsurface) and also the deepest layers (e.g., burrowing shrimp Upogebia pusilla; Petagna). The experiment was conducted in summer 2000 (August-September) at three different sites on an intertidal flat in Mediterranean Sea. In the field, predators were excluded using cages, placed on the surface of the sediment. It was predicted that under the cages, (i) abundances of animals would increase and (ii) species composition of assemblages would change as an effect of the exclusion of predators. Potential artefacts caused by the cages were controlled using partial cages (PC). Composition of organic matter and porosity were also analysed under PC and in natural controls to test whether the presence of cages would change sediment characteristics on the surface. At the end of the experiment, there were no significant differences in the composition of organic matter between PC and the undisturbed sediment (UC). After 8 weeks from the beginning of the experiment, exclusion of predators enhanced the abundance of U. pusilla. There were, however, no clear-cut changes in the species composition of macrofaunal assemblages and densities of animals did not increase under the cages. Indeed, some animals (Oligochaeta, Brania oculata, and Tanais dulongii) were less abundant under the cages (EC) than outside (PC and UC). We propose that predation might play a role in regulating interspecific relationships among some surface deposit-feeders and the burrowing shrimp U. pusilla.     

Analysis of competitive interactions between the limpets Patella depressa Pennant and Patella vulgata L. on the northern coast of Portugal

Inter- and intraspecific competitive interactions among the species Patella depressa Pennant and P. vulgata L. were analysed on the northern coast of Portugal, where both species co-occur in similar proportions. Increased (×2, ×4), normal and decreased (×1/2 intraspecific only) densities of limpets were used to test the effects of competition on the growth and mortality of the limpets, and competitive interactions between the different species. Fenced plots of 25×25 cm enclosing marked limpets at various densities were set up at a mid tidal level on the shore. Twelve treatments with three replicates of different combinations of densities and species were established. Mortality was recorded every 15 days and length was measured monthly during the 5 months of the experiment. At the end of the experiment, limpets were collected for biometry, sex determination and gonad staging. Both species of grazing molluscs showed increased mortality and reduced size and weight in increased density treatments. Limpets in decreased density treatments showed lower mortality and higher size. There were no significant differences between the effect of P. vulgata on P. depressa and the effect of P. depressa on P. vulgata on mortality and length. There were, however, decreases in weight due to intraspecific effects for both species. For P. vulgata, interspecific effects were much less than intraspecific effects and for P. depressa, the interspecific effects depended on density. The analysis of the gonad stage at the end of the experiment showed that the lower stages of development were most prevalent at increased densities and the occurrence of neuter individuals only in increased densities single species plots corroborated the evidence of a stronger intraspecific interaction. The availability of food was indirectly assessed by determination of chlorophyll concentration with spectrophotometric analysis of rock chips. Microalgal food assessment in this experiment, however, did not show any clear trends. The results were compared with previous studies of competition in grazing molluscs and discussed in relation to the limits of distribution of P. vulgata on the Portuguese coast.     

Cold tolerance of Littorinidae from southern Africa: intertidal snails are not constrained to freeze tolerance

All intertidal gastropods for which cold tolerance strategies have been assessed have been shown to be freeze tolerant. Thus, freeze tolerance is considered an adaptation to the intertidal environment. We investigated the cold tolerance strategies of three species of subtropical and temperate snails (Gastropoda: Littorinidae) to determine whether this group is phylogenetically constrained to freeze tolerance. We exposed dry acclimated and wet rehydrated snails to low temperatures to determine temperature of crystallisation (Tc), lower lethal temperature and LT₅₀ and to examine the relationship between ice formation and mortality. Tc was lowest in dry Afrolittorina knysnaensis (–13.6±0.4 °C), followed by dry Echinolittorina natalensis (–10.9±0.2 °C) and wet A. knysnaensis (–10.2±0.2 °C). The Tc of both A. knysnaensis and E. natalensis increased with rehydration, whereas Tc of dry and wet Afrolittorina africana did not differ (–9.6±0.2 and –9.0±0.2 °C respectively). Wet snails of all species exhibited no or low survival of inoculative freezing, whereas dry individuals of A. knysnaensis could survive subzero temperatures above –8 °C when freezing was inoculated with ice. In the absence of external ice, Afrolittorina knysnaensis employs a freeze-avoidance strategy of cold tolerance, the first time this has been reported for an intertidal snail, indicating that there is no family-level phylogenetic constraint to freeze tolerance. Echinolittorina natalensis and A. africana both showed pre-freeze mortality and survival of some internal ice formation, but were not cold hardy in any strict sense.     

Limits to the distribution of the southern African Ice Rat (Otomys sloggetti): thermal physiology or competitive exclusion?

1. We test the hypothesis that the thermal physiology of Otomys sloggetti restricts its distribution to altitudes above 2000 m, below which it does not occur.
2. The thermal physiology of O . sloggetti is typical of rodents from warm/hot areas: the thermoneutral zone is relatively broad and lies between 26 and 28 °C; the upper limit of the thermoneutral zone can be extended by 4 °C through the use of hyperthermia; basal metabolic rate concurs with the allometrically predicted value; the lower critical temperature is high; minimal thermal conductance is 21% higher than the allometrically predicted value; and the capacity for noradrenaline-induced non-shivering thermogenesis is 19% lower than the allometrically predicted value.
3. Evaporative water loss and water flux rates are high and typical of mesic-adapted rodents.
4. The absence of O . sloggetti from warmer, lower-lying habitats cannot be attributed to its thermal physiology. We suggest that non-physiological factors, such as interspecific competition with Otomys irroratus , are more likely to restrict the distribution of O . sloggetti .     

Evidences of habitat displacement between two common soft-bottom SW Atlantic intertidal crabs

The intertidal burrowing crab Chasmagnathus granulatus Dana is the dominant species in soft sediments and vegetated intertidal areas along the SW Atlantic estuaries (southern Brazil 28°S to the northern Argentinean Patagonia 41°S) where it produces dense and extensive burrowing beds. The mud crab Cyrtograpsus angulatus Dana coexists with Ch. granulatus in this area, but it also inhabits areas to the south (northern and central Argentinean Patagonia). A survey covering both areas showed that C. angulatus rarely live in burrows when coexisting with Ch. granulatus, but form large burrowing beds when not coexisting with Ch. granulatus. When both species coexisted, burrowing beds of C. angulatus are restricted to sandy-muddy areas. Only rarely are burrows of C. angulatus found within Ch. granulatus beds. However, when Ch. granulatus were experimentally excluded within their burrowing beds, new settlers of C. angulatus made burrows and maintained them until they reached large size. Paired (inside and outside Ch. granulatus burrowing bed) sampling during high tide using beach nets showed that C. angulatus rarely venture inside the Ch. granulatus crab beds. Other field experiments showed that adults Ch. granulatus always displace C. angulatus from burrows. Furthermore, in several sites located south of the limit of distribution of Ch. granulatus at the Patagonian coast, soft bare intertidals are dominated by burrowing beds of C. angulatus mixed with the congener C. altimanus Dana. Together, these evidences suggest that the mud crab C. angulatus is displaced from soft bottom areas by the burrowing crab Ch. granulatus. It is an example of competitive exclusion through aggressive interference in soft-bottom habitats when the shared resource is the access to sediment surface, a two-dimensional well-defined resource.     

Why spacing behavior does not stabilize density in cyclic populations of microtine rodents

Summary There are several published hypotheses that consider spacing behavior to be a significant factor causing the multiannual density fluctuations characteristic of some microtine rodent populations. Recent modeling efforts have concluded, however, that spacing behavior should have a stabilizing rather than a destabilizing effect on population dynamics. Why doesn't spacing behavior stabilize these cyclic populations? We argue that while spacing behavior does have a stabilizing influence on population dynamics by limiting the number of breeding individuals, reproduction continues and population size is not limited in an asymptotic manner. Rather, microtine social organization produces demographic changes within a population that allow density cycles to occur under certain conditions. Using a simulation model, we demonstrate that in a strongly seasonal environment populations with low density dependence in reproduction will cycle whereas populations with high density dependence in reproduction will have relatively stable densities. Given such complicating factors as the annual species nature of microtine rodents, occasionally intense predation, and the tendency for territoriality to break down during the non-breeding season, individuals with low density dependence in reproduction will always be able to invade and eventually dominate populations with high density dependence in reproduction, regardless of the resulting destabilization of population dynamics.     

Recovery of photosynthesis after exposure of intertidal algae to osmotic and temperature stresses: comparative studies of species with differing distributional limits

Summary This study examines possible relationships between stress tolerance by marine algae and distributions of these species. The ability to recover photosynthetic activity following dehydration or temperature treatments was the assay used to evaluate stress tolerance, and Porphyra perforata, Rhodoglossum affine, Gelidium coulteri, and Smithora naiadum differed in thresholds of tolerance, even though plants were collected from low tidal sites.Limits of dehydration tolerance were well correlated with limits of tidal distribution for these species. Additionally, other high tidal species tolerated severe dehydration while subtidal and low tidal species were sensitive to dehydration. High tidal individuals of P. perforata were also more tolerant of dehydration than were low tidal thalli of P. perforata.Limits of high or low temperature tolerance were not well correlated with tidal elevation for any groups of algae studied. However, cold-tolerant species had more northerly extensions, and warm-tolerant species had more southerly distributions. Thus, differential tolerance to temperature extremes may be an important influence for latitudinal ranges of species.By comparing the experimentally determined thresholds of stress with distributions of species, we test the role of stress in influencing photosynthesis and ultimately distributions of marine algae.     

Simple replacement of violaxanthin by zeaxanthin in LHC-II does not cause chlorophyll fluorescence quenching

Recently, a mechanism for the energy-dependent component (qE) of non-photochemical quenching (NPQ), the fundamental photo-protection mechanism in green plants, has been suggested. Replacement of violaxanthin by zeaxanthin in the binding pocket of the major light harvesting complex LHC-II may be sufficient to invoke efficient chlorophyll fluorescence quenching. Our quantum chemical calculations, however, show that the excited state energies of violaxanthin and zeaxanthin are practically identical when their geometry is constrained to the naturally observed structure of violaxanthin in LHC-II. Therefore, since violaxanthin does not quench LHC-II, zeaxanthin should not either. This theoretical finding is nicely in agreement with experimental results obtained by femtosecond spectroscopy on LHC-II complexes containing violaxanthin or zeaxanthin.     

ER stress does not cause upregulation and activation of caspase-2 to initiate apoptosis

A recent report claimed that endoplasmic reticulum (ER) stress activates the ER trans-membrane receptor IRE1α, leading to increased caspase-2 levels via degradation of microRNAs, and consequently induction of apoptosis. This observation casts caspase-2 into a central role in the apoptosis triggered by ER stress. We have used multiple cell types from caspase-2-deficient mice to test this hypothesis but failed to find significant impact of loss of caspase-2 on ER-stress-induced apoptosis. Moreover, we did not observe increased expression of caspase-2 protein in response to ER stress. Our data strongly argue against a critical role for caspase-2 in ER-stress-induced apoptosis.     

Theory does not meet experiment: transient dynamics changes patterns of exclusion in an intraguild predation system

Population Ecology - Although empirical verifications of ecological theory are essential for the advance of our understanding of ecosystems functioning, they are often hard to obtain or even...     

Experimental evidence that evolutionary relatedness does not affect the ecological mechanisms of coexistence in freshwater green algae

The coexistence of competing species depends on the balance between their fitness differences, which determine their competitive inequalities, and their niche differences, which stabilise their competitive interactions. Darwin proposed that evolution causes species' niches to diverge, but the influence of evolution on relative fitness differences, and the importance of both niche and fitness differences in determining coexistence have not yet been studied together. We tested whether the phylogenetic distances between species of green freshwater algae determined their abilities to coexist in a microcosm experiment. We found that niche differences were more important in explaining coexistence than relative fitness differences, and that phylogenetic distance had no effect on either coexistence or on the sizes of niche and fitness differences. These results were corroborated by an analysis of the frequency of the co‐occurrence of 325 pairwise combinations of algal taxa in > 1100 lakes across North America. Phylogenetic distance may not explain the coexistence of freshwater green algae.     

Intraperitoneal injection of synthetic bacterial lipopeptides does not cause a rise in circulating inflammatory cytokines

Abstract The production of TNF-α, IL-1 and IL-6 was measured in mice after i.p. injections of the synthetic bacterial lipopeptide Pam₃Cys-Ala-Gly, a potent macrophage and B cell activator in vitro. Only minor amounts of IL-6 and no TNF-α were detectable in the serum of mice injected with 10 or 100 μg of Pam₃Cys-Ala-Gly. Lipopeptide concentrations up to 1000 μg failed to induce IL-1, and TNF-α production and serum IL-6 levels were only slightly elevated. In contrast to Pam₃Cys-Ala-Gly, i.p. injections of LPS were accompanied by high levels of the inflammatory cytokines TNF-α, IL-1 and IL-6 suggesting fundamental differences of the mode of action of these two substances when applied in vivo.     

Why dominants do not consistently attain high mating and reproductive success: A review of longitudinal Japanese macaque studies

There is wide interest in the effects of reproductive biology, mating partner preference, and rank on mating success (MS) and reproductive success (RS) in primates. In particular, theory stresses importance on the mechanisms for attaining RS. Most theory hedges on competitive ability and priority of access to resources, whether they be food or estrous females. However, the majority of data used in favor of such hypotheses come from relatively short-term studies. We review these hypotheses based on long-term data from provisioned and unprovisioned populations of Japanese macaques. Neither MS nor RS were consistently attained by high-ranking males and females. For males, female choice and mating partner preference is seen to over-ride most male-male competitive behaviors likely to affect MS and RS through priority of access to estrous females. Long-term mating patterns driven largely by female partner preferences, results in decreasing MS and RS for older higher-ranking males. The long-term trend for females to prefer less familiar or novel partners results in higher MS and RS for younger, middle-ranking males. The effects of this vary according to troop size and the duration of male tenure. For females, no consistent trend was recognized for rank related RS in either provisioned or unprovisioned troops. Non-reproductive mating may provide differential benefit to high-ranking females for access to limited food resources in some habitats but overall the relationship was inconclusive. Distribution and defendability of food resource, rather than provisioning per se may be more important.     

A massive systematic infection of Encephalitozoon cuniculi genotype III in mice does not cause clinical signs

Encephalitozoon cuniculi genotype III disseminated intensively into most of the organs in all strains of mice, followed by a chronic infection with massive microsporidia persistence in immunodeficient mice and a partial decrease in C57Bl/6 mice. Treatment with 0.2 mg Albendazole/mouse/day temporarily reduces the number of affected organs in immunocompetent C57Bl/6 mice, but not in CD4^−/− and CD8^−/− mice. The application of medication temporarily decreased the spore burden at least by one order of magnitude in all groups.These results demonstrate that the E. cuniculi genotype III infection had a progressive course and surprisingly, Albendazole treatment had only a minimal effect. The E. cuniculi genotype III spore burden in individual organs reached up to 10⁸ or 10⁹ in immunocompetent or immunodeficient mice, respectively; however, these mice did not demonstrate any obvious clinical signs of microsporidiosis, and the immunodeficient mice survived longer. Our findings clearly show that the survival of mice does not correspond to spore burden, which provides new insight into latent microsporidiosis from an epidemiological point of view.