Pressure treatment of <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> in low-moisture environments

We investigated the influence of cell hydration on the ability of Saccharomyces cerevisiae CBS 1171 to withstand extreme hydrostatic pressure in order to determine the mechanisms involved in cell resistance. Hydration conditions were modified in two different ways. We first modulated the chemical potential of water by adding glycerol in cell suspensions. Another procedure consisted in dehydrating cells aerobically and immersing them in perfluorooctane, an innocuous hydrophobic liquid used as a pressure-transmitting medium, prior to pressure treatments. This original method made it possible to transmit isostatic pressure to yeast powders without changing the initial water activity (a _w) level at which cells had been equilibrated. The a _w ranged between 0.11 and 0.99. Pressure treatments were applied at levels of up to 600 MPa for 10 min, 24 h, and 6 days. The dehydration of cells was found to strongly limit, or even prevent, cell inactivation under pressure. Notably, cells suspended in a water–glycerol mixture with a _w levels of 0.71 or below were completely protected against all pressure treatments. Moreover, cells dehydrated aerobically survived for 6 days at 600 MPa even when a _w levels were relatively high (up to 0.94). We highlighted the crucial role of water content in determining cellular damage under pressure. When water is available in a sufficient amount, high pressure induces membrane permeabilization, causing uncontrolled mass transfers that could lead to death during a prolonged holding under pressure. Possible mechanisms of membrane permeabilization are discussed.     

Unfolding studies of tissue transglutaminase

Activation of tissue transglutaminase by calcium involves a conformational change which allows exposition of the active site to the substrate via movements of domains 3 and 4 that lead to an increase of the inter-domain distance. The inhibitor GTP counteracts these changes. Here we investigate the possible existence of non-native conformational states still compatible with the enzyme activity produced by chemical and thermal perturbations. The results indicate that chemical denaturation is reversible at low guanidine concentrations but irreversible at high concentrations of guanidine. Indeed, at low guanidine concentrations tissue TG-ase exists in a non-native state which is still affected by the ligands as in the native form. In contrast, thermal unfolding is always irreversible, with aggregation and protein self-crosslinkage in the presence of calcium. DSC thermograms of the native protein in the absence of ligands consist of two partly overlapped transitions, which weaken in the presence of calcium and merge together and strengthen in the presence of GTP. Overall, the present work shows, for the first time, the reversible denaturation of a TG-ase isoenzyme and suggests the possibility that also in in vivo, the enzyme may acquire non-native conformations relevant to its patho-physiological functions.     

Senescence-Accelerated Mouse (SAM) with Special References to Neurodegeneration Models,SAMP8 and SAMP10 Mice

The SAM strains, a group of related inbred strains consisting of senescence-prone inbred strains (SAMP) and senescence-resistant inbred strains (SAMR), have been successfully developed by selective inbreeding of the AKR/J strain of mice donated by the Jackson laboratory in 1968. The characteristic feature of aging common to the SAMP and SAMR is accelerated senescence and normal aging, respectively. Furthermore, SAMP and SAMR strains of mice manifest various pathobiological phenotypes spontaneously. Among SAMP strains, SAMP8 and SAMP10 mice show age-related behavioral deterioration such as deficits in learning and memory, emotional disorders (reduced anxiety-like behavior and depressive behavior) and altered circadian rhythm associated with certain pathological, biochemical and pharmacological changes. Here, the previous and recent literature on SAM mice are reviewed with an emphasis on SAMP8 and SAMP10 mice. A spontaneous model like SAM with distinct advantages over the gene-modified model is hoped by investigators to be used more widely as a biogerontological resource to explore the etiopathogenesis of accelerated senescence and neurodegenerative disorders.     

Significant ecosystem-wide effects of the swiftly spreading invasive freshwater bivalve <Emphasis Type="Italic">Limnoperna</Emphasis> <Emphasis Type="Italic">fortunei</Emphasis>

Since its introduction in South America around 1990, the freshwater Asian mussel Limnoperna fortunei has been shown to strongly interact with several components of the local biota. However, investigation of its ecosystem-wide effects was hindered by (1) difficulties associated with evaluation of its densities over large spatial scales and (2) scarcity of pre-invasion environmental data. The present survey overcomes these shortcomings and addresses the question whether Limnoperna’s impact on the ecosystem-wide scale is measurable and significant. On the basis of diver-collected bottom samples, we estimated the overall density of this mussel in a reservoir (Embalse de Río Tercero, Argentina), where Limnoperna is present since 1998 and analyzed changes in several water-column properties before and after the invasion. The 47 km² reservoir hosts around 45 billion mussels; at these densities, a volume equivalent to that of this water body can potentially be filtered by the bivalves every 2–3 days. Data collected regularly since 1996 indicate that after the invasion water transparency increased, and suspended matter, chlorophyll a, and primary production decreased significantly, with strong changes occurring in the area with highest mussel densities. Our results indicate that the ecosystem-wide impacts of Limnoperna are generally comparable to those described in Europe and North America for another invasive mussel—Dreissena polymorpha. However, given Limnoperna’s wider tolerance limits, its influence on newly invaded water bodies, potentially including Europe and North America, will probably be stronger.     

Developing an <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated genetic transformation for a selenium-hyperaccumulator <Emphasis Type="Italic">Astragalus racemosus</Emphasis>

Agrobacterium tumefaciens strain LBA4404 containing the plasmid pBI121, carrying the reporter gene uidA and the kanamycin resistance gene nptII, was used for gene transfer experiments in selenium (Se)-hyperaccumulator Astragalus racemosus. The effects of kanamycin on cell growth and division and acetosyringone on transformation efficiency were evaluated. The optimal concentration of kanamycin that could effectively inhibit cell growth and division in non-transgenic tissues was 50 mg l⁻¹ and thus all putative transgenic plants were obtained on induction medium containing 50 mg l⁻¹ kanamycin. The verification of transformants was achieved by both histochemical GUS assay and PCR amplification of nptII gene. Southern blot analysis was performed to further confirm that transgene nptII was stably integrated into the A. racemosus genome. A transformation frequency of approximately 10% was achieved using this protocol, but no beneficial effect from the addition of acetosyringone (50 μM) was observed. This transformation system will be a useful tool for future studies of genes responsible for Se-accumulation in A. racemosus.     

Speed encoding in correlation motion detectors as a consequence of spatial structure

For animals to carry out a wide range of detection, recognition and navigation tasks, visual motion signals are crucial. The encoding of motion information has therefore, attracted much attention in the experimental and computational study of brain function. Two main alternative mechanisms have been proposed on the basis of behavioural and physiological experiments. On one hand, correlation-type and motion energy detectors are simple and efficient in the design of their basic mechanism but are tuned to temporal frequency rather than to speed. On other hand, gradient-type motion detectors directly represent an estimate of speed, but may require more demanding processing mechanisms. We demonstrate here how the temporal frequency dependence observed for sine-wave gratings can disappear for less constrained stimuli, to be replaced by responses reflecting speed for stimuli like square waves when a phase-sensitive detection mechanism is employed. We conclude from these observations that temporal frequency tuning is not necessarily a limitation for motion vision based on correlation detectors, and more generally demonstrate in view of the typical Fourier composition of natural scenes, that correlation detectors operating in such environments can encode image speed. In the context of our results, we discuss the implications of the loss of phase sensitivity inherent in using a linear system approach to describe neural processing.     

Seasonal dynamics of zooplankton in a shallow eutrophic,man-made hyposaline lake in Delhi (India): role of environmental factors

Old Fort Lake, a small (1.6 ha), shallow, and recreational water body in Delhi (India) was studied through monthly surveys in two consecutive years (January, 2000–December, 2001). Precipitation is the major source of water for this closed basin lake. In addition, ground water is used for replenishing the lake regularly. This alkaline, hyposaline hard water lake contains very high ionic concentration, especially of nitrates. Based on overall ionic composition, this lake can be categorized as chloride–sulfate alkaline waters with the anion sequence dominated by SO₄ ²⁻ > Cl⁻ > HCO₃ ⁻, and the cations by Mg⁺⁺ > Ca⁺⁺. The overall seasonal variability in physicochemical profile was largely regulated by the annual cycle of evaporation and precipitation, whereas the ground water largely influences its water quality. The lake exhibited phytoplankton-dominated turbid state due to dominance of the blue green alga, Microcystis aeruginosa. The persistent cyanobacterial blooms and the elevated nutrient levels are indicative of the cultural eutrophication of the lake. This study focuses on the relative importance of eutrophic vis-à-vis hyposaline conditions in determining the structure and seasonal dynamics of zooplankton species assemblages. A total of 52 zooplankton species were recorded and rotifers dominated the community structure qualitatively as well as quantitatively. The genus Brachionus comprised a significant component of zooplankton community with B. plicatilis as the most dominant species. The other common taxa were B. quadridentatus, B. angularis, Lecane grandis, L. thalera, L. punctata, Mesocyclops sp., and Alona rectangula. Multivariate data analysis techniques, Canonical Correspondence Analysis (CCA) along with Monte Carlo Permutation Tests were used to determine the minimum number of environmental factors that could explain statistically significant (P < 0.05) proportions of variation in the species data. The significant variables selected by CCA were NH₃–N followed by percent saturation of DO, COD, SS, BOD, NO₂–N, rainfall, silicates, and PO₄–P. The results indicate that the seasonal succession patterns of the zooplankton species were largely controlled by physicochemical factors related directly or indirectly to the process of eutrophication, whereas hyposaline conditions in the lake determined the characteristic species composition. Guest Editors: J. John & B. Timms Salt Lake Research: Biodiversity and Conservation—Selected papers from the 9th Conference of the International Society for Salt Lake Research     

Dietary Enrichment with Medium Chain Triglycerides (AC-1203) Elevates Polyunsaturated Fatty Acids in the Parietal Cortex of Aged Dogs: Implications for Treating Age-Related Cognitive Decline

Dogs demonstrate an age-related cognitive decline, which may be related to a decrease in the concentration of omega-3 polyunsaturated fatty acids (n-3 PUFA) in the brain. Medium chain triglycerides (MCT) increase fatty acid oxidation, and it has been suggested that this may raise brain n-3 PUFA levels by increasing mobilization of n-3 PUFA from adipose tissue to the brain. The goal of the present study was to determine whether dietary MCT would raise n-3 PUFA concentrations in the brains of aged dogs. Eight Beagle dogs were randomized to a control diet (n = 4) or an MCT (AC-1203) enriched diet (n = 4) for 2 months. The animals were then euthanized and the parietal cortex was removed for phospholipid, cholesterol and fatty acid determinations by gas-chromatography. Dietary enrichment with MCT (AC-1203) resulted in a significant increase in brain phospholipid and total lipid concentrations (P < 0.05). In particular, n-3 PUFA within the phospholipid, unesterified fatty acid, and total lipid fractions were elevated in AC-1203 treated subjects as compared to controls (P < 0.05). Brain cholesterol concentrations did not differ significantly between the groups (P > 0.05). These results indicate that dietary enrichment with MCT, raises n-3 PUFA concentrations in the parietal cortex of aged dogs.     

Removal of selenite from wastewater using microbial fuel cells

Simultaneous electricity generation and selenium removal was evaluated in single-chamber microbial fuel cells (MFCs) with acetate and glucose as carbon sources. Power output was not affected by selenite up to 125 mg l⁻¹ with glucose as substrate. Coulombic efficiencies of MFCs with glucose increased from 25% to 38% at 150 mg Se l⁻¹. About 99% of 50 and 200 mg Se l⁻¹ selenite was removed in 48 and 72 h for MFCs fed with acetate and glucose, respectively, demonstrating the potential of using MFC technology for Se remediation.     

Quantification of indole-3-acetic acid from plant associated <Emphasis Type="Italic">Bacillus</Emphasis> spp. and their phytostimulatory effect on <Emphasis Type="Italic">Vigna radiata</Emphasis> (L.)

Sixteen Bacillus strains isolated from rhizosphere, histoplane and phyllosphere of different plant species were identified by 16S rDNA gene sequencing and evaluated for in vitro auxin production as well as growth stimulation of Vigna radiata (L.) Wilczek. Auxin production by Bacillus spp. in L-broth medium supplemented with 1,000 μg ml⁻¹ L-tryptophan ranges from 0.60 to 3.0 μg IAA ml⁻¹ as revealed by gas chromatography and mass spectrometric (GC–MS) analysis. Rhizospheric isolates exhibit relatively more IAA synthesis than histoplane and phyllosphere isolates. Plant microbe interaction experiments conducted under gnotobiotic conditions recorded 55.55, 46.46 and 46.20% increase in shoot length with Bacillus megaterium MiR-4, B. pumilus NpR-1 and B. subtilis TpP-1, respectively, over control. Bacillus inoculations also increased shoot fresh weight with B. megaterium MiR-4 (60.94%) and B. pumilus NpR-1 (37.76%). Highly significant positive correlation between auxin production analyzed by GC–MS and shoot length (r = 0.687**, P = 0.01) and shoot fresh weight (r = 0.703**, P = 0.01) was noted under gnotobiotic conditions. Similarly, significant correlation was also found between auxin production by Bacillus spp. (GC–MS analysis) and different growth parameters such as shoot length (r = 0.495*, P = 0.05), number of pods (r = 0.498*, P = 0.05) and grain weight (r = 0.537*, P = 0.05) at full maturity under natural wire house conditions. Results showed that auxin production potential of plant associated Bacillus spp. can be effectively exploited to enhance the growth and yield of V. radiata.