β-d-Glucosidases and related enzymic activities in pig kidney

1. The beta-glucosidase activity of pig kidney is located in the unsedimentable fraction of the cell and is not associated with the lysosomes. 2. The enzyme is active towards beta-d-glucosides, beta-d-galactosides, beta-d-xylosides and alpha-l-arabinosides. 3. These activities could not be separated by gel electrophoresis, gel filtration or DEAE-cellulose chromatography. 4. Response to inhibitors, heat-denaturation and competitive substrates suggests that a single active site is responsible for all four activities. 5. Two forms of the enzyme were found to occur either separately or together in kidneys of pigs from several different breeds. 6. Electro-focusing experiments show these to have a small difference in isoelectric point (4.9 and 5.1), and gel filtration gives an approximate molecular weight of 50000 for both forms. 7. The characteristics of these two enzymes are compared.     

How pregnancy can affect autoimmune diseases progression?

Autoimmune disorders are characterized by tissue damage, caused by self-reactivity of different effectors mechanisms of the immune system, namely antibodies and T cells. Their occurrence may be associated with genetic and/or environmental predisposition and to some extent, have implications for fertility and obstetrics. The relationship between autoimmunity and reproduction is bidirectional. This review only addresses the impact of pregnancy on autoimmune diseases and not the influence of autoimmunity on pregnancy development. Th17/Th1-type cells are aggressive and pathogenic in many autoimmune disorders and inflammatory diseases. The immunology of pregnancy underlies the role of Th2-type cytokines to maintain the tolerance of the mother towards the fetal semi-allograft. Non-specific factors, including hormonal changes, favor a switch to Th2-type cytokine profile. In pregnancy Th2, Th17/Th2 and Treg cells accumulate in the decidua but may also be present in the mother’s circulation and can regulate autoimmune responses influencing the progression of autoimmune diseases.     

How does temperature play a role in the storage of extracellular vesicles?

Extracellular vesicles (EVs) contain specific proteins, lipids, and nucleic acids that can be passed to other cells as signal molecules to alter their function. However, there are many problems and challenges in the conversion and clinical application of EVs. Storage and protection of EVs is one of the issues that need further research. To adapt to potential clinical applications, this type of problem must be solved. This review summarizes the storage practices of EVs in recent years, and explains the impact of temperature on the quality and stability of EVs during storage based on current research, and explains the potential mechanisms involved in this effect as much as possible.     

How do changes in temperature during growth affect leaf pigment composition and photosynthesis in Zea mays genotypes differing in sensitivity to low temperature?

The changes in photosynthetic activity and composition of pigments induced by changes in temperature were examined in the third leaf of three chilling-tolerant and three chilling-sensitive genotypes of maize (Zea mays L.). The plants were grown under a controlled environment at a photon flux density of 550 mol m^-2 s^-1, a 12 h photoperiod and at a suboptimal temperature of 14/12 C (day/night) until the full expansion of the third leaf. After this treatment, the chilling-tolerant genotypes, when compared with the sensitive ones, displayed a higher photosynthetic activity, a higher content of chlorophyll (Chl) a+b, a higher Chl a/b ratio, a larger total carotenoid pool size as well as a different carotenoid composition. When temperature was subsequently increased to 24/22 C for 3 d the composition of the pigments changed, but the chilling-sensitive genotypes, while adjusting their lower Chl a/b ratio and their different carotenoid composition, were unable to adjust their lower content of chlorophyll, their smaller total carotenoid pool size or their lower photosynthetic performance. Moreover, while the chilling-tolerant genotypes converted the most part of zeaxanthin to violaxanthin in the xanthophyll cycle, the chilling-sensitive genotypes retained high amounts of zeaxanthin. The changes in pigment composition that occurred over the 3 d at 24/22 °C were largely conserved when the plants were returned to 14/12 °C, but photosynthetic activity decreased and zeaxanthin accumulated again. The results suggest that the capability of the chilling-tolerant genotypes, when compared with the sensitive ones, to retain high amounts of pigments and to form a competent photosynthetic apparatus at low temperature is the basis for their more vigorous growth in cool climates.     

How will increased temperature and nutrient enrichment affect primary producers in sub‐Arctic streams?

1. Spring‐fed streams, with temperatures ranging from 7.1 to 21.6 °C, in an alpine geothermal area in SW Iceland were chosen to test hypotheses on the effects of nutrients and temperature on stream primary producers. Ammonium nitrate was dripped into the lower reaches of eight streams, with higher reaches being used as controls, during the summers of 2006 and 2007. Dry mass of larger primary producers, epilithic chlorophyll a and biovolumes of epilithic algae were measured. 2. Bryophyte communities were dominated by Fontinalis antipyretica, and biomass was greatest in the warmest streams. Jungermannia exsertifolia, a liverwort, was found in low densities in few samples from cold streams but this species was absent from the warmest streams. 3. Nutrient enrichment increased the biomass of bryophytes significantly in warm streams. No effects of the nutrient addition were detected on vascular plants. The biomass of larger filamentous algae (mainly Cladophora spp.) was significantly increased by nutrient enrichment in cold streams but reduced by nutrients in warm streams. Thalloid cyanobacteria (Nostoc spp.) were not affected by nutrients in cold streams but decreased with nutrient addition in warm streams. Epilithic algal chlorophyll a was increased by nutrients in all streams and to a greater extent in 2007 than in 2006. Nutrient addition did not affect the epilithic chlorophyll a differently in streams of different temperatures. 4. There were small differential effects of nutrients, influenced by pH and conductivity, on different epilithic algal groups. 5. As global temperatures increase, animal husbandry and perhaps crop agriculture are likely to increase in Iceland. Temperature will directly influence the stream communities, but its secondary effects, manifested through agricultural eutrophication, are likely to be much greater.     

How do floating aquatic weeds affect wetland conservation and development? How can these effects be minimised?

The most important floating aquatic weeds (FAWs) are Eichhornia crassipes, Salvinia molesta and Pistia stratiotes. E. crassipes and P. stratiotes reproduce sexually. All three species reproduce asexually. E. crassipes and S. molesta have particularly high growth rates. All can form dense mats and growth rates are increased by high nutrient levels and temperatures. Spread between continents and watersheds is largely the result of human activities. Spread within watersheds is mostly via floating propagules. FAWs are known to affect water resource management, the continued existence of human riverine and wetland communities, and conservation of biodiversity. Waterways can be blocked, and the efficiency of irrigation and hydro generation impaired. People are affected by reduction of the fish catch, inability to travel by boat and consequent isolation from gardens, markets and health services, and also changes in populations of vectors of human and animal diseases. Biodiversity can be reduced and conservation value affected. It is proposed that rational application of physical, chemical and biological control of FAWs, and reduction of nutrient input should be part of every strategy for the sustainable management of wetlands.     

How noise in force fields can affect the structural refinement of protein models?

Structural refinement of predicted models of biological macromolecules using atomistic or coarse‐grained molecular force fields having various degree of error is investigated. The goal of this analysis is to estimate what is the probability for designing an effective structural refinement based on computations of conformational energies using force field, and starting from a structure predicted from the sequence (using template‐based or template‐free modeling), and refining it to bring the structure into closer proximity to the native state. It is widely believed that it should be possible to develop such a successful structure refinement algorithm by applying an iterative procedure with stochastic sampling and appropriate energy function, which assesses the quality (correctness) of protein decoys. Here, an analysis of noise in an artificially introduced scoring function is investigated for a model of an ideal sampling scheme, where the underlying distribution of RMSDs is assumed to be Gaussian. Sampling of the conformational space is performed by random generation of RMSD values. We demonstrate that whenever the random noise in a force field exceeds some level, it is impossible to obtain reliable structural refinement. The magnitude of the noise, above which a structural refinement, on average is impossible, depends strongly on the quality of sampling scheme and a size of the protein. Finally, possible strategies to overcome the intrinsic limitations in the force fields for impacting the development of successful refinement algorithms are discussed. Proteins 2012. © 2011 Wiley Periodicals, Inc.     

Coexistence and coevolution in fluctuating environments: can the storage effect evolve?

Both theoretical and empirical work have shown that the temporal storage effect can promote coexistence. However, the storage effect depends on temporally fluctuating demographic rates, such as interannually variable germination rates. Because variable demographic rates often reduce fitness, we asked how a storage effect might evolve. Using a model of competing annual plants, we find that variable germination (and hence a storage effect) can evolve only if germination is positively correlated with high-fecundity years (predictive germination) or if one species has limited ability to evolve. Outside of these conditions, the storage effect is evolutionarily unstable: if two species were thrown together with traits that would permit a storage effect, they would coevolve constant germination rates and the storage effect would vanish. Our results suggest that for the temporal storage effect to be widespread, either germination must commonly be positively correlated with later growth or fecundity or community assembly must reflect biogeographic processes more than they do coevolution.     

How do water transport and water storage differ in coniferous earlywood and latewood?

The goal of this research project was to determine the water transport behaviour of earlywood versus latewood in the trunk of 21-year-old Douglas-fir [Pseudostuga menziesii (Mirb.) Franco] trees. Specific conductivity (k(s)) and the vulnerability of xylem to embolism were measured on a single growth ring and in a subset of earlywood and latewood samples within the same ring. Earlywood/latewood ratio, trunk water potential (Psi) and relative water content (RWC) were used to predict differences in conductivities and vulnerability to embolism. Earlywood has about 11 times the k(s) of latewood, and up to 90% of the total flow occurred through the earlywood. Earlywood's vulnerability to embolism followed the same trend as that of the whole wood, with 50% loss of conductivity at -2.2 MPa (P(50)). Latewood was more vulnerable to embolism than earlywood at high Psi, but as Psi decreased, the latewood showed very little further embolism, with a P(50) <-5.0 MPa. The lowest trunk Psi estimated in the field was about -1.4 MPa, indicating that latewood and earlywood in the field experienced about 42% and 16% loss of k(s), respectively. The higher vulnerability to embolism in latewood than in earlywood at field Psi was associated with higher water storage capacity (21.8% RWC MPa(-1) versus 4.1% RWC MPa(-1), latewood and earlywood, respectively). The shape of the vulnerability curve suggests that air seeding through latewood may occur directly through pores in the margo and seal off at lower pressure than earlywood pores.     

How do additives affect enzyme activity and stability in nonaqueous media?

The catalytic activities of lyophilized powders of alpha-chymotrypsin and Candida antarctica lipase were found to increase 4- to 8-fold with increasing amounts of either buffer salts or potassium chloride in the enzyme preparation. Increasing amounts of sorbitol in the chymotrypsin preparation produced a modest increase in activity. The additives are basically thought to serve as immobilization matrices, the sorbitol being inferior because of its poor mechanical properties.Besides their role as supports, the buffer species were indispensable for the transesterification activity of chymotrypsin because they prevented perturbations of the pH during the course of the reaction. Hence, increasing amounts of buffer species yielded a 100-fold increase in transesterification activity. Effects of pH changes were not as predominant in the peptide synthesis and the lipase-catalyzed reactions.Immobilization of the protease on celite resulted in a remarkable improvement of transesterification activity as compared to the suspended protease, even in the absence of buffer species. Immobilization of the lipase caused a small improvement of activity. The activity of the immobilized enzymes was further enhanced 3-4 times by including increasing amounts of buffer salts in the preparation.The inclusion of increasing amounts of sodium phosphate or sorbitol to chymotrypsin rendered the catalyst more labile against thermal inactivation. The denaturation temperature decreased with 7 degrees C at the highest content of sodium phosphate, as compared to the temperature obtained for the denaturation of the pure protein. The apparent enthalpy of denaturation increased with increasing contents of the additives. The enhancement of hydration level and flexibility of the macromolecule upon addition of the compounds partly provides the explanation for the observed results. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 54: 67-76, 1997.     

Does dietary arsenic and mercury affect cutaneous bleeding time and blood lipids in humans?

Fish species may contain considerable amounts of trace elements, such as selenium (Se), arsenic (As), and mercury (Hg). The present study investigated the relationships between dietary intake of these elements and cutaneous bleeding time and blood lipids in 32 healthy volunteers. For 6 wk, one group (n=11) consumed approx 250 g Se-rich fish daily, providing them with an average Se intake of 115±31 μg Se/d, Hg intake of 18±8 μg/d, and As intake of 806±405 μg/d, all values analyzed in 4-d duplicate food collections. To study the effect of Se alone, one group (n=11) included Se-rich bread in their normal diet, giving them a Se intake (135±25 μg/d) that was comparable to the fish group. A control group (n=10) ate their normal diet, providing 77±25 gmg Se/d, 3.1±2.5 μg Hg/d, and 101±33 μg As/d. The dietary As load strongly correlated both with bleeding times and changes in bleeding times (r=0.48,p<0.01 andr=0.54,p<0.002, respectively). Dietary Hg showed a positive correlation with LDL-cholesterol (r=0.55,p<0.01), whereas dietary Hg in the fish group showed a strong negative relationship with HDL-cholesterol (r=?0.76,p<0.01). Selenium seemed to have only a modest effect on bleeding time. Our results suggest that mercury and arsenic from fish may be factors contributing to or modifying some of the known effects of fish ingestion.     

How does storage affect the quality and quantity of organic carbon in sewage for use in the bioremediation of acidic mine waters?

Pit lakes (abandoned flooded mine pits) represent a potentially valuable water resource. However, acid mine drainage (AMD) generation due to mining activities often results in pit lake waters with low pH, high sulphate and dissolved metal concentrations. Sulphate reduction-based bioremediation offers tremendous scope for removal of acidity and metals from pit lake water. In this study, the effect of storing sewage on its carbon quality for bioremediation of acidic pit lake water was studied. In addition, the effectiveness of labile organic carbon (lactic acid and ethanol) on SRB activity was tested. Bioremediation experiments were performed in controlled and replicated microcosms with acidic (pH 2.2) water from a pit lake by addition of stored (3 years at 4 °C) sewage for stimulation of sulphate reducing bacteria (SRB) activity. This sewage had been previously used successfully in remediating to pH 7 water from this pit lake. The initial aim was to test the sewage at lower doses (18 and 28 g/L) and in a pulsed addition (over 5 weeks). Bioremediation efficacy was evaluated by measuring pit lake water pH increase, redox potential decrease, and acidity and sulphate removal. Though the stored sewage had retained a very similar high total organic carbon (TOC) equivalent to prior to storage, it failed to increase dissolved organic carbon (DOC) levels in pit lake water. Microcosms amended with doubled doses of sewage and an extended remediation time still failed to demonstrate any substantial improvement in water quality, other than a small amount of sulphate reduction and direct neutralisation by the sewage. In order to determine if low DOC concentrations in sewage were the cause of the bioremediation failure, labile organic carbon (LOC), consisting of 50:50 (w/w) lactic acid and ethanol, was added to all microcosm treatments at concentrations of 3000, 6000 and 9000 mg/L. After LOC addition, water quality improved with effective removal of acidity, sulphate and metals in the lowest carbon concentration (3000 mg/L). However, 6000 and 9000 mg/L LOC concentrations showed a delay in response due to the increased acidity associated with the lactic acid addition. The experiments showed that pulsed dosing of carbon simply slowed the commencement of remediation but it was ultimately able to reach the same effectiveness as the equivalent quantity added all at once. Prolonged storage of sewage leads to loss of LOC. In situ pit lake remediations which aim to make use of sewage as the main carbon source will need to factor in the storage time required to obtain sufficient sewage for the treatment into the design. Pulsing may help reduce issues with storage or supplementation with LOC may need to be considered. Results highlight that LOC is a more useful indicator of material effectiveness compared to a simple measures of TOC.     

How do sheep affect plant communities and arthropod populations in temperate grasslands?

Grasslands being used in sheep farming systems are managed under a variety of agricultural production, recreational and conservational objectives. Although sheep grazing is rarely considered the best method for delivering conservation objectives in seminatural temperate grasslands, the literature does not provide unequivocal evidence on the impact of sheep grazing on pasture biodiversity. Our aim was therefore to review evidence of the impacts of stocking rate, grazing period and soil fertility on plant communities and arthropod populations in both mesotrophic grasslands typical of agriculturally improved areas and in native plant communities. We therefore conducted a literature search of articles published up to the end of the year 2010 using ‘sheep’ and ‘grazing’ as keywords, together with variables describing grassland management, plant community structure or arthropod taxa. The filtering process led to the selection of 48 articles, with 42 included in the stocking rate dataset, 9 in the grazing period dataset and 10 in the soil fertility dataset. The meta-analysis did not reveal any significant trends for plant species richness or plant community evenness along a wide stocking rate gradient. However, we found frequent shifts in functional groups or plant species abundance that could be explained by the functional properties of the plants in the community. The meta-analysis confirmed that increasing soil fertility decreased plant species richness. Despite the very limited dataset, plant species richness was significantly greater in autumn-grazed pastures than in ungrazed areas, which suggests that choosing an appropriate grazing period would be a promising option for preserving biodiversity in sheep farming systems. Qualitative review indicated that low grazing intensity had positive effects on Orthoptera, Hemiptera (especially phytophagous Auchenorrhyncha) and, despite a diverse range of feeding strategies, for the species richness of Coleoptera. Lepidoptera, which were favoured by more abundant flowering plants, also benefited from low grazing intensities. Spider abundance and species richness were higher in ungrazed than in grazed pastures. In contrast, there are insufficient published studies to draw any firm conclusions on the benefits of late grazing or stopping fertilization on insect diversity, and no grounds for including any of this information in decision support tools at this stage.     

How do grazers affect periphyton heterogeneity in streams?

The effects of grazing by stream invertebrates on algal biomass and spatial heterogeneity were tested experimentally in flow-through microcosms with natural substrates (rocks). One experiment tested the effects of fixed densities of three species of grazers (the caddisfly Allomyia sp. and two mayflies, Epeorus deceptivus and Baetis bicaudatus) on periphyton. Baetis was tested with and without chemical cues from fish predators, which reduced grazer foraging activity to levels similar to the less mobile mayfly (Epeorus). Mean algal biomass (chlorophyll a; chl a) was reduced in grazer treatments compared to ungrazed controls, but there were no differences among grazer treatments. Algal heterogeneity (Morisita index) increased with grazer mobility, with the highest heterogeneity occurring in the Baetis-no fish treatment (most mobile grazer) and the lowest in the caddisfly treatment (most sedentary grazer). A second experiment used a three factorial design, and tested whether initial resource distribution (homogeneous vs. heterogeneous), Baetis density (high vs. low) and fish odor (present vs. absent) affected grazer impact on algal resources. Abundances of Baetis and chl a on individual rocks were recorded to explore the mechanisms responsible for the observed distributions of algae. Initial resource heterogeneity was maintained despite being subjected to grazing. Mean chl a was highest in controls, as in experiment I, and effects of Baetis on algal biomass increased with grazer density. There were no fish effects on algal biomass and no effects of grazer density or fish on algal heterogeneity. At the scale of individual rocks Baetis was unselective when food was homogeneously distributed, but chose high-food rocks when it was heterogeneously distributed. Results of these mechanistic experiments showed that Baetis can track resources at the scale of single rocks; and at moderate densities mobile grazers could potentially maintain periphyton distributions observed in natural streams.     

Does storage time influence yield and agar properties in the tropical agarophyte Gracilaria cornea?

The agar yield and quality characteristics of Gracilaria cornea from Yucatán, Mexico, werestudied during 18 months of storage. Biomass wasstored at a temperature of 22.1 ± 0.9 ^°Cand humidity of 59.8±3.6%. The agar contentvaried erratically, but the average value waspractically constant over the storage period with an average of 20.1 ± 1.5±. Gel strength, gelling and meltingtemperatures were negatively affected by the totalstorage time. No significant changes were found duringthe first five months for these characteristics withmean values of 1134 ± 57 g cm^-2, 40.8 ±0.4 ^°C and 91.2 ± 0.9 ^°Crespectively. Agar degradation was evident after thefifth month and accounted for a 17± loss in gelstrength and 7± in gelling and meltingtemperatures. Nevertheless, gel strength valuesremained around 930 ± 23 g cm^-2 with nosignificant changes until the end of the storageperiod. The decrease in gel strength showed asignificant relationship with decrease in3,6-anhydrogalactose but not variation in sulphatecontent. This was probably due to agar hydrolysiscaused by enzymatic processes of endogenous and/ormicrobial origin. These results suggest that thetropical G. cornea had a similar resistance todegradation during storage to that observed for G. chilensis, a cold water species. Agar qualityand yield in G. cornea after one and a half yearof storage are within the range of food grade agar.     

How does the nature of living and dead roots affect the residence time of carbon in the root litter continuum?

Root litter transformation is an important determinant of the carbon cycle in grassland ecosystems. Litter quality and rhizosphere activity are species-dependent factors which depend on the attributes of the dead and living roots respectively. These factors were tested, using non-disturbed soil monoliths ofDactylis glomerata L. andLolium perenne L. monocultures.¹³C-labelled root litter from these monoliths was obtained from a first stand of each crop, cultivated under veryδ ¹³C-depleted atmospheric CO₂ (S1). In a factorial design,¹³C-labelled root litter of each species was submitted to a second, non¹³C-labelled, living stand of each species (S2). Carbon derived from S1 and from S2 was measured during an 18-month incubation in the root phytomass and in three particulate organic matter fractions (POM). The decay rate of each particle size fraction was fitted to the experimental data in a mechanistic model of litter transformation, whose outputs were mineralisation and stabilisation of the litter-C. Few differences were found between species, in the amount and biochemical composition of the initial root litter, butDactylis roots showed a greater C:N ratio, a lower mean root diameter and a greater specific root length compared toLolium. A transient accumulation of litter residues arose successively in POM fractions of decreasing particle size. The litter-continuum hypothesis was validated, i.e. that the attributes of the compartments (C:N, chemical composition and residence time) depended mainly on their particle size. The S1 species influenced the rate of litter decay while the S2 species controlled the efficiency of litter-C stabilisation versus mineralisation:Dactylis litter decomposed faster andLolium rhizosphere allowed a greater proportion of litter C stabilisation. Discussions focus on the processes responsible of species strategy in relation with the morphological root traits, and the implication of strategy diversity for rich grassland communities.     

How can we bring together empiricists and modellers in functional biodiversity research?

Improving our understanding of biodiversity and ecosystem functioning and our capacity to inform ecosystem management requires an integrated framework for functional biodiversity research (FBR). However, adequate integration among empirical approaches (monitoring and experimental) and modelling has rarely been achieved in FBR. We offer an appraisal of the issues involved and chart a course towards enhanced integration. A major element of this path is the joint orientation towards the continuous refinement of a theoretical framework for FBR that links theory testing and generalization with applied research oriented towards the conservation of biodiversity and ecosystem functioning. We further emphasize existing decision-making frameworks as suitable instruments to practically merge these different aims of FBR and bring them into application. This integrated framework requires joint research planning, and should improve communication and stimulate collaboration between modellers and empiricists, thereby overcoming existing reservations and prejudices. The implementation of this integrative research agenda for FBR requires an adaptation in most national and international funding schemes in order to accommodate such joint teams and their more complex structures and data needs.     

What can be observed in real time PCR and when does it show?

Journal of Mathematical Biology - Real time, or quantitative, PCR typically starts from a very low concentration of initial DNA strands. During iterations the numbers increase, first essentially by...