Activity-dependent phosphorylation of SNAP-25 in hippocampal organotypic cultures.

Synaptosomal-associated protein of 25 kDa (SNAP-25) is thought to play a key role in vesicle exocytosis and in the control of transmitter release. However, the precise mechanisms of action as well as the regulation of SNAP-25 remain unclear. Here we show by immunoprecipitation that activation of protein kinase C (PKC) by phorbol esters results in an increase in SNAP-25 phosphorylation. In addition, immunochemical analysis of two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels shows that SNAP-25 focuses as three or four distinct spots in the expected range of molecular weight and isoelectric point. Changing the phosphorylation level of the protein by incubating the slices in the presence of either a PKC agonist (phorbol 12,13-dibutyrate) or antagonist (chelerythrine) modified the distribution of SNAP-25 among these spots. Phorbol 12,13-dibutyrate increased the intensity of the spots with higher molecular weight and lower isoelectric point, whereas chelerythrine produced the opposite effect. This effect was specific for regulators of PKC, as agonists of other kinases did not produce similar changes. Induction of long-term potentiation, a property involved in learning mechanisms, and production of seizures with a GABA(A) receptor antagonist also increased the intensity of the spots with higher molecular weight and lower isoelectric point. This effect was prevented by the PKC inhibitor chelerythrine. We conclude that SNAP-25 can be phosphorylated in situ by PKC in an activity-dependent manner.

Producció de biopolímers amb cultius bacterians mixtes. Comparació de tres reactors discontinus seqüencials (SBR) amb diferents condicions operacionals

La producció de biopolímers (polihidroxialcanoats (PHA) i substàncies polimèriques extracel·lulars (EPS)) a nivell industrial, resulta una nova àrea d’investigació que recull diverses disciplines, entre elles les Ciències Ambientals. Aquest projecte final de carrera amb el títol: “Producció de biopolímers amb cultius bacterians mixtes”, s’ha desenvolupat sota la supervisió de la directora de projecte Dra. María Eugenia Suárez Ojeda del Departament d’Enginyeria Química de la Universitat Autònoma de Barcelona (UAB) i s’ha dut a terme per l’estudiant Jordi Pérez i Forner de la Llicenciatura de Ciències Ambientals, Facultat de Ciències de la UAB, en el Departament d’Enginyeria Química de la mateixa universitat. L’objectiu d’aquest projecte ha estat produir biopolímers simultàniament amb l’eliminació de fòsfor i matèria orgànica en aigües residuals per obtenir un residu final amb un alt valor afegit. Aquests biopolímers reuneixen les característiques necessàries per a poder competir amb els plàstics convencionals i així, reduir l’elevat consum del petroli i la generació de residus no biodegradables. En aquest projecte s’ha dut a terme la posta en marxa d’un reactor discontinu seqüencial (SBR) per a l’acumulació de biopolímers amb cultius bacterians mixtes. Diferents investigadors han estudiat que aquests tipus de cultius bacterians arriben a nivells de fins el 53-97% [Pijuan et al., 2009] de contingut de biopolímers a la biomassa, sometent als microorganismes a diferents situacions d’estrés ja sigui per dèficit de nutrients o per variacions en les fases de feast-famine (festí-fam). Durant el projecte, s’ha realitzat el monitoratge del reactor alimentat amb una aigua sintètica, elaborada en el laboratori, amb les característiques d’un aigua residual provinent de la industria làctica. S’ha sotmès als microorganismes a diferents condicions operacionals, una d’elles amb limitació de fòsfor com a nutrient i una tercera condició amb una variació a les fases feast-famine. D’altra banda, com a segon objectiu, s’ha analitzat el contingut de biopolímers a la biomassa de dos SBRs més, del grup de recerca Bio-GLS del Departament d’Enginyeria Química de la UAB, alimentats amb diferents fonts de carboni, glicerol i àcids grassos de cadena llarga (AGCLL), per observar les influències que té el tipus de substrat en l’acumulació de biopolímers. Els resultats obtinguts en la primera part d’aquest projecte han estat similars als resultats d’altres investigadors [Pijuan et al., 2009; Guerrero et al., 2012]. S’ha determinat que sotmetre als microorganismes a situacions d’estrés té un efecte directe pel que fa a l’acumulació de biopolímers. També s’ha observat com al mateix temps que acumulaven aquests compostos, els microorganismes desenvolupaven la seva tasca de depurar l’aigua residual, obtenint al final del cicle una aigua amb un baix contingut en matèria orgànica i altres contaminants com amoni i fòsfor, en aquest cas. En la segona part del projecte, s’ha observat com el tipus de substrat té un efecte directe pel que fa a l’acumulació de biopolímers i també a l’activitat metabòlica dels microorganismes. Per tant, s’ha conclòs que la producció de biopolímers mitjançant la depuració d’aigües residuals es una via d’investigació molt prometedora pel que fa als resultats obtinguts. Alhora que es tracta un residu, s’obté una producte residual amb un alt valor afegit que pot ser utilitzat per la producció de bioplàstics 100% biodegradables.

Analysis of variance of primary data on plant growth analysis

Plant growth analysis presents difficulties related to statistical comparison of growth rates, and the analysis of variance of primary data could guide the interpretation of results. The objective of this work was to evaluate the analysis of variance of data from distinct harvests of an experiment, focusing especially on the homogeneity of variances and the choice of an adequate ANOVA model. Data from five experiments covering different crops and growth conditions were used. From the total number of variables, 19% were originally homoscedastic, 60% became homoscedastic after logarithmic transformation, and 21% remained heteroscedastic after transformation. Data transformation did not affect the F test in one experiment, whereas in the other experiments transformation modified the F test usually reducing the number of significant effects. Even when transformation has not altered the F test, mean comparisons led to divergent interpretations. The mixed ANOVA model, considering harvest as a random effect, reduced the number of significant effects of every factor which had the F test modified by this model. Examples illustrated that analysis of variance of primary variables provides a tool for identifying significant differences in growth rates. The analysis of variance imposes restrictions to experimental design thereby eliminating some advantages of the functional growth analysis.

08008-004 Rathbun Lake Final Report

The Rathbun Land and Water Alliance and partners have undertaken a highly effective approach to water quality protection through the Rathbun Lake Special Project. This approach is achieving a significant reduction in the sediment and phosphorus that impair water quality in Rathbun Lake and its tributaries as a result of the targeted application of best management practices (BMPs) for priority land in the watershed. This project application proposes to assist landowners to apply BMPs that will reduce sediment and phosphorus delivery from priority land in one targeted sub-watershed as part of the Rathbun Lake Special Project. Features of this project are: (1) use of geographic information system (GIS) analysis to identify priority land that requires BMPs; (2) assistance for landowners to apply BMPs on 1,450 acres that will reduce the annual delivery of sediment by 2,160 tons and phosphorus by 8,210 pounds; (3) evaluation of the benefits from BMP application using GIS analysis and water quality monitoring; and (4) watershed outreach activities that encourage landowners to apply BMPs for priority land to protect water quality.

07043-017 North Fork Maquoketa Final Report

The North Fork Maquoketa River Headwaters (NFMRH) has been identified as impaired by nutrients, episodic slugs of ammonia and sediment. The NFMRH TMDL plan calls for a "phasing approach" to managing water quality when the origin is non-point source contaminants. This project will address phase 1 using a performance reward program for targeted cooperators to improve environmental index scores using cost-share, EQIP practices and flexible management alternatives. Pre-project assessments suggest that rewards should target refined management of erosion-prone fields and farms with livestock populations, which contribute to the P and N loads responsible for fertilizing filamentous algae blooms that depress dissolved oxygen concentrations in the NFMRH. The Phosphorus Index, Soil Conditioning Index and cornstalk nitrate test will be used by producers to determine effective alternatives, such as no-till planting, to reduce nutrient and sediment delivery. These evironmental indexes will be especially useful for livestock producers in the livestock dense watershed. This project will extend a NRCS-sponsored Conservation Innovation Grant currently offered to producers in the Coffee Creek sub-watershed to a three-year, watershed-wide effort that will be necessary to make significant improvements in environmental management.

Growth, yield, water and effluent quality in ponds with different management during tambaqui juvenile production

The objective of this work was to evaluate the effect of pond management on fish feed, growth, yield, survival, and water and effluent quality, during tambaqui (Colossoma macropomum) juvenile production. Fish were distributed in nine 600 m² earthen ponds, at a density of 8 fish per m²; the rearing period was 60 days. Three different pond management were applied: limed and fertilized (LimFer), limed (Lim), and natural (Nat). Fish were fed with a commercial ration containing 34% crude protein three times daily. There were no significant differences in fish growth or yield. Three main items found in tambaqui stomach were insect, zooplankton and ration, without a significant difference among treatments in proportion. Alkalinity, hardness, and CO2 were greater in LimFer and Lim ponds. Chlorophyll a, transparency, ammonia, nitrite, temperature, and dissolved oxygen of pond water were not significantly different among treatments. Biochemical oxygen demand, total phosphorus, orthophosphate, ammonia, and nitrite were significantly greater in effluents from LimFer ponds. Pond fertilization should be avoided, because growth and yield were similar among the three pond management systems tested; besides, it produces a more impacting effluent.

Effects of usual nutrient intake and vitamin D status on markers of bone turnover in Swiss adolescents.

OBJECTIVE: To evaluate the effects of nutrient intake and vitamin D status on markers of type I collagen formation and degradation in adolescent boys and girls. DESIGN: Cross-sectional study. SETTING: Canton of Vaud, West Switzerland. SUBJECTS: A total of 92 boys and 104 girls, aged 11-16 y. Data were collected on height, weight, pubertal status (self-assessment of Tanner stage), nutrient intake (3-day dietary record) and fasting serum concentration of 25-hydroxyvitamin D (25OHD), and markers of collagen formation (P1NP) and degradation (serum C-terminal telopeptides: S-CTX). RESULTS: Tanner stage was a significant determinant of P1NP in boys and girls and S-CTX in girls. Of the nutrients examined, only the ratio of calcium to phosphorus (Ca/P) was positively associated with P1NP in boys, after adjustment for pubertal status. 25OHD decreased significantly at each Tanner stage in boys. Overall, 15% of boys and 17% of girls were identified as being vitamin D insufficient (serum 25OHD <30 nmol/l), with the highest proportion of insufficiency at Tanner stage 4-5 (29%) in boys and at Tanner stage 3 (24%) in girls. A significant association was not found between 25OHD and either bone turnover marker, nor was 25OHD insufficiency associated with higher concentrations of the bone turnover markers. CONCLUSIONS: The marked effects of puberty on bone metabolism may have obscured any possible effects of diet and vitamin D status on markers of bone metabolism. The mechanistic basis for the positive association between dietary Ca/P ratio and P1NP in boys is not clear and may be attributable to a higher Ca intake per se, a critical balance between Ca and P intake or higher dairy product consumption. A higher incidence of vitamin D insufficiency in older adolescents may reflect a more sedentary lifestyle or increased utilisation of 25OHD, and suggests that further research is needed to define their requirements. SPONSORSHIP: Nestec Ltd and The Swiss Foundation for Research in Osteoporosis.

Method for evaluation of root hairs of common bean genotypes

The objective of this work was to test a simple method for root hair evaluation of 21 common bean (Phaseolus vulgaris) genotypes, most of them used in breeding programs in Brazil. Hairs of basal and primary roots of 5-day old seedlings, produced on germination paper with no phosphorus addition, were visually evaluated by a rating scale after staining with 0.05% trypan blue. The method reveals variability among the genotypes, and the standard error of the mean is relatively low.

New isolates of ectomycorrhizal fungi and the growth of eucalypt

The objective of this work was to evaluate the efficiency of ectomycorrhizal isolates on root colonization, phosphorus uptake and growth of Eucalyptus dunnii seedlings. Inocula of ten ectomycorrhizal isolates of Chondrogaster angustisporus, Hysterangium gardneri, Pisolithus spp., and Scleroderma spp. were aseptically produced in a peat-vermiculite mixture supplemented with liquid culture medium. Plants grew in a similar substrate supplemented with macro-and micro-nutrients; treatments were randomly distributed in a greenhouse. After three months, seedlings inoculated with three isolates - UFSC-Sc68 (Scleroderma sp.), UFSC-Ch163 (Chondrogaster angustisporus), and UFSC-Pt188 (Pisolithus microcarpus) - had a phosphorus shoot content and a shoot dry matter higher or equivalent to those of noninoculated controls which had been fertilized with a 16-fold phosphorus amount. These isolates were selected for new studies for establishing inoculum production techniques, in order to be applied in reforestation programmes under nursery and field conditions.

Latex production with depolymerizing compounds of actin cytoskeleton in rubber trees

The objective of this work was to assess stimulated latex flow from rubber trees (Hevea brasiliensis) with saturated macrolide (latrunculin A), 1, 5, and 10% potassium iodide in 2% methylcellulose compared with 0.3% ethylene in 2% methylcellulose (check) and 2% methylcellulose (blank). Latex output and contents of pure rubber, total solid, sucrose, inorganic phosphorus, thiol, and Mg2+ were measured. The treatments containing 1% KI or saturated macrolide increased latex yields compared to the blank with 2% methylcellulose alone. The 1% KI or saturated macrolide treatments were equal to that of 0.3% ethylene check treatment. However, 5 and 10% KI were harmful to bark of rubber trees, even caused prolonged tapping panel dryness.

Dynamic Modelling of Material Flows and Sustainable Resource Use: Case Studies in Regional Metabolism and Space Life Support Systems

Sustainable resource use is one of the most important environmental issues of our times. It is closely related to discussions on the 'peaking' of various natural resources serving as energy sources, agricultural nutrients, or metals indispensable in high-technology applications. Although the peaking theory remains controversial, it is commonly recognized that a more sustainable use of resources would alleviate negative environmental impacts related to resource use. In this thesis, sustainable resource use is analysed from a practical standpoint, through several different case studies. Four of these case studies relate to resource metabolism in the Canton of Geneva in Switzerland: the aim was to model the evolution of chosen resource stocks and flows in the coming decades. The studied resources were copper (a bulk metal), phosphorus (a vital agricultural nutrient), and wood (a renewable resource). In addition, the case of lithium (a critical metal) was analysed briefly in a qualitative manner and in an electric mobility perspective. In addition to the Geneva case studies, this thesis includes a case study on the sustainability of space life support systems. Space life support systems are systems whose aim is to provide the crew of a spacecraft with the necessary metabolic consumables over the course of a mission. Sustainability was again analysed from a resource use perspective. In this case study, the functioning of two different types of life support systems, ARES and BIORAT, were evaluated and compared; these systems represent, respectively, physico-chemical and biological life support systems. Space life support systems could in fact be used as a kind of 'laboratory of sustainability' given that they represent closed and relatively simple systems compared to complex and open terrestrial systems such as the Canton of Geneva. The chosen analysis method used in the Geneva case studies was dynamic material flow analysis: dynamic material flow models were constructed for the resources copper, phosphorus, and wood. Besides a baseline scenario, various alternative scenarios (notably involving increased recycling) were also examined. In the case of space life support systems, the methodology of material flow analysis was also employed, but as the data available on the dynamic behaviour of the systems was insufficient, only static simulations could be performed. The results of the case studies in the Canton of Geneva show the following: were resource use to follow population growth, resource consumption would be multiplied by nearly 1.2 by 2030 and by 1.5 by 2080. A complete transition to electric mobility would be expected to only slightly (+5%) increase the copper consumption per capita while the lithium demand in cars would increase 350 fold. For example, phosphorus imports could be decreased by recycling sewage sludge or human urine; however, the health and environmental impacts of these options have yet to be studied. Increasing the wood production in the Canton would not significantly decrease the dependence on wood imports as the Canton's production represents only 5% of total consumption. In the comparison of space life support systems ARES and BIORAT, BIORAT outperforms ARES in resource use but not in energy use. However, as the systems are dimensioned very differently, it remains questionable whether they can be compared outright. In conclusion, the use of dynamic material flow analysis can provide useful information for policy makers and strategic decision-making; however, uncertainty in reference data greatly influences the precision of the results. Space life support systems constitute an extreme case of resource-using systems; nevertheless, it is not clear how their example could be of immediate use to terrestrial systems.

09018-011 Rathbun Lake Final Report

The Rathbun Land and Water Alliance and partners have undertaken a highly effective approach to water quality protection through the Rathbun Lake Special Project. This approach is achieving a significant reduction in the sediment and phosphorus that impair water quality in Rathbun Lake and its tributaries as a result of the targeted application of best management practices (BMPs) for priority land in the watershed. This application proposes to assist landowners to apply BMPs that will reduce sediment and phosphorus delivery from priority land in targeted sub-watersheds as part of the Rathbun Lake Special Project. Features of this project are: (1) use of geographic information system (GIS) analysis to identify priority land that requires BMPs; (2) assistance for landowners to apply BMPs for 4,000 acres that will reduce the annual delivery of sediment by 6,000 tons and phosphorus by 20,000 pounds; (3) evaluation of the benefits from BMP application using GIS analysis and water quality monitoring; and (4) watershed outreach activities that encourage landowners to apply BMPs for priority land to protect water quality.

09008-005 Hewitt Creek Final Report

The primary goal of the Hewitt Creek watershed council is to have Hewitt-Hickory Creek removed from the Iowa impaired waters (303d) list. Hewitt Creek watershed, a livestock dense 23,005 acre sub-watershed of the Maquoketa River Basin, is 91.2% agricultural and 7.5% woodland. Since 2005, sixty-seven percent of 84 watershed farm operations participated in an organized watershed improvement effort using a performance­ based watershed management approach, reducing annual sediment delivery to the stream by 4,000 tons. Watershed residents realize that water quality improvement efforts require a long-term commitment in order to meet their watershed improvement goals and seek funding for an additional five years to continue their successful watershed improvement project. Cooperators will be provided incentives for improved environmental performance, along with incentives and technical support to address feedlot runoff issues and sub-surface nitrate-nitrogen loss. The Phosphorus Index, Soil Conditioning Index and cornstalk nitrate test will be used by producers as measures of performance to refine nutrient and soil loss management and to determine effective alternatives to reduce nutrient and sediment delivery. Twenty-five livestock operations will improve feedlot runoff control systems and five sub-surface bioreactors will be installed to reduce nitrate delivery from priority tile-drained fields. The Hewitt Creek council will seek additional cost-share funding for high-cost feedlot runoff control structures, sediment control basins and stream bank stabilization projects.

The effects of polycyclic aromatic hydrocarbons and heavy metals on terrestrial annelids in urban soils

The effect of soil contamination by polycyclic aromatic hydrocarbons (PAH) and heavy metals on earthworms and enchytraeids was studied in urban parks, in Brno, Czech Republic. In spring and autumn 2007, annelids were collected and soil samples taken in lawns along transects, at three different distances (1, 5 and 30 m) from streets with heavy traffic. In both seasons, two parks with two transects each were sampled. Earthworms were collected using the electrical octet method. Enchytraeids were extracted by the wet funnel method from soil cores. All collected annelids were counted and identified. Basic chemical parameters and concentrations of 16 PAH, Cd, Cu, Pb, and Zn were analysed from soil from each sampling point. PAH concentrations were rather low, decreasing with the distance from the street in spring but not in autumn. Heavy metal concentrations did not decrease significantly with increasing distance. Annelid densities did not significantly differ between distances, although there was a trend of increase in the number of earthworms with increasing distance. There were no significant correlations between soil content of PAH or heavy metals and earthworm or enchytraeid densities. Earthworm density and biomass were negatively correlated with soil pH; and enchytraeid density was positively correlated with soil phosphorus.

Low-P tolerance mechanisms and differential gene expression in contrasting wheat genotypes

The objectives of this study were to determine low-P tolerance mechanisms in contrasting wheat genotypes and to evaluate the association of these mechanisms to differential gene expression. Wheat seedlings of cultivars Toropi (tolerant to low-P availability) and Anahuac (sensitive) were evaluated. Seedlings were hydroponically grown in the absence or presence of P (1.0 mmol L-1) during three different time periods: 24, 120 and 240 hours. Free phosphate (Pi) and total P contents were measured in shoots and roots. The experiment's design was in randomized blocks with three replicates, each formed by ten plants. The relative expression of genes encoding the malate transporter TaALMT1 and the transcription factor PTF1 was evaluated. Phosphorus starvation beyond ten days increased the expression of TaALMT1 only in 'Toropi'. PTF1's expression was early induced in both genotypes under P starvation, but remained significant after ten days only in 'Toropi'. Shoot Pi concentration in 'Toropi' was independent from P availability; under starvation, 'Toropi' favored the maintenance of shoot Pi concentration. The low-P tolerance of Toropi cultivar at initial growth stages is mainly due to its ability to maintain constant the Pi shoot level.