IN VITRO EFFECTS OF FUMONISIN B1 ALONE AND COMBINED WITH DEOXYNIVALENOL OR ZEARALENONE ON PORCINE GRANULOSA CELL PROLIFERATION AND STEROIDOGENESIS

Fumonisins, zearalenone and trichothecenes, such as deoxynivalenol (DON) and T-2 toxin, are the major Fusarium mycotoxins occurring throughout the world in cereal grains and animal feeds. Direct ovarian effects of fumonisin B1 (FB1) and its interaction with DON or \u3b1-zearalenol (\u3b1-ZOL), zearalenone major active metabolite, have so far not been investigated. Thus, the goal of this thesis was to determine if FB1, alone or combined with DON or \u3b1-ZOL, can impair swine reproductive activity via affecting granulosa cell function. To this aim, two different studies were designed. In the first study the effects of FB1 alone and combined with DON or \u3b1-ZOL on granulosa cell proliferation were evaluated. Porcine granulosa cells from small ovarian follicles (1-5 mm) were cultured for 2 days in 5% fetal bovine serum and 5% porcine serum-containing medium followed by 2 days in serum-free medium containing FB1 at various doses (0, 0.01, 0.4, 10 and 14 \u3bcM) and combinations. At the end of the experiments, numbers of granulosa cells were determined using a Coulter counter. The results revealed that FB1 had inhibitory effects on granulosa cell proliferation at doses 65 10 \u3bcM. DON (3.4 \u3bcM) strongly inhibited (by 80%; P < 0.0001) granulosa cell proliferation and no significant difference was detected in combination with FB1 (10 \u3bcM). \u3b1-ZOL (9.4 \u3bcM) showed a stimulatory effect (P < 0.01) on granulosa cell numbers, even when treated in combination with FB1 (10 \u3bcM). In the second study the effects of FB1 alone and combined with DON or \u3b1-ZOL on granulosa cell steroid production and gene expression were investigated. Porcine granulosa cells from small follicles (1-5 mm) were cultured as described above. At the end of the experiments, concentrations of progesterone and estradiol in culture medium were determined by radioimmunoassays. Real-time RT-PCR was used to elucidate the effects of FB1 on gene expression of P450scc (CYP11A1) and aromatase (CYP19A1). All doses of FB1 (i.e., 0.01, 0.4, 10 and 14 \ub5M) had no significant effect on estradiol production, whereas FB1 showed a stimulatory effect on progesterone production induced by FSH plus insulin-like growth factor-I (IGF-I) at 10 and 14 \ub5M. \u3b1-ZOL (9.4 \ub5M) increased (P < 0.0001) FSH plus IGF-I-induced progesterone production by 51%. Combination of FB1 with \u3b1-ZOL resulted in an increase of progesterone production (91%; P < 0.0001) that was significantly higher than that induced by either Fusarium mycotoxin alone. DON drastically inhibited (by 74%; P < 0.0001) progesterone production and FB1 had little effect on this response. \u3b1-ZOL had no effect on FSH plus IGF-I-induced estradiol production, whereas decreased (P < 0.05) estradiol production when co-treated with FB1. DON (3.4 \ub5M) strongly inhibited (by 67%; P < 0.0001) estradiol production and no difference was detected in combination with FB1 (10 \ub5M). FB1 (10 \ub5M) had no effect on granulosa cell CYP19A1 mRNA abundance, whereas decreased (by 23%; P < 0.0001) granulosa cell CYP11A1 mRNA abundance induced by FSH plus IGF-I. In conclusion, the present thesis indicates that FB1 has direct effects on porcine granulosa cell proliferation, steroid production and gene expression and provides information on the interactions between FB1 and DON or \u3b1-ZOL in granulosa cells. These interactions and direct ovarian effects should be considered in swine reproductive failures

A simple modeling approach to study the regional impact of a Mediterranean forest isoprene emission on anthropogenic plumes

Research during the past decades has outlined the importance of biogenic isoprene emission in tropospheric chemistry and regional ozone photo-oxidant pollution. The first part of this article focuses on the development and validation of a simple biogenic emission scheme designed for regional studies. Experimental data sets relative to Boreal, Tropical, Temperate and Mediterranean ecosystems are used to estimate the robustness of the scheme at the canopy scale, and over contrasted climatic and ecological conditions. A good agreement is generally found when comparing field measurements and simulated emission fluxes, encouraging us to consider the model suitable for regional application. Limitations of the scheme are nevertheless outlined as well as further on-going improvements. In the second part of the article, the emission scheme is used on line in the broader context of a meso-scale atmospheric chemistry model. Dynamically idealized simulations are carried out to study the chemical interactions of pollutant plumes with realistic isoprene emissions coming from a Mediterranean oak forest. Two types of anthropogenic sources, respectively representative of the Marseille (urban) and Martigues (industrial) French Mediterranean sites, and both characterized by different VOC/NOx are considered. For the Marseille scenario, the impact of biogenic emission on ozone production is larger when the forest is situated in a sub-urban configuration (i.e.&nbsp;downwind distance TOWN-FOREST <30km, considering an advection velocity of 4.2 m.s^-1). In this case the enhancement of ozone production due to isoprene can reach +37% in term of maximum surface concentrations and +11% in term of total ozone production. The impact of biogenic emission decreases quite rapidly when the TOWN-FOREST distance increases. For the Martigues scenario, the biogenic impact on the plume is significant up to TOWN-FOREST distance of 90km where the ozone maximum surface concentration enhancement can still reach +30%. For both cases, the importance of the VOC/NO_x ratio in the anthropogenic plume and its evolution when interacting with the forest emission are outlined. In complement to real case studies, this idealized approach can be particularly useful for process and sensitivity studies and constitutes a valuable tool to build regional ozone control strategies

Alkaloid-Containing Plants Poisonous to Cattle and Horses in Europe

Alkaloids, nitrogen-containing secondary plant metabolites, are of major interest to veterinary toxicology because of their occurrence in plant species commonly involved in animal poisoning. Based on epidemiological data, the poisoning of cattle and horses by alkaloid-containing plants is a relatively common occurrence in Europe. Poisoning may occur when the plants contaminate hay or silage or when forage alternatives are unavailable. Cattle and horses are particularly at risk of poisoning by Colchicum autumnale (meadow saffron), Conium maculatum (poison hemlock), Datura stramonium (jimson weed), Equisetum palustre (marsh horsetail), Senecio spp. (ragwort and groundsel) and Taxus baccata (European yew). This review of poisonous alkaloid-containing plants describes the distribution of these plants, conditions under which poisoning occurs, active toxic principles involved and subsequent clinical signs observed

Detection of multiple mycotoxin occurrences in soy animal feed by traditional mycological identification combined with molecular species identification

Soy products are a main component of animal feed. Because mycotoxins may harm farm animals, undermining productivity and health, a mycological and toxigenic screening was carried out on 36 batches used in animal feed, collected in 2008, 2009 and 2010 in Italy. The investigated mycoflora of a subset of soy seed (n = 6) suggested that Aspergillus spp. and Fusarium spp. frequently colonize soy seeds. Aflatoxins, fumonisins and deoxynivalenol were detected in 88.9%, 72.2% and 30.6% of samples, respectively. Co-occurrence of at least two toxins was observed in 72% of cases. The molecular analysis of the Fusarium spp. population identified Fusarium verticillioides as potential producers of fumonisins, but no known deoxynivalenol producers were detected. It is suggested that the widespread presence of toxins can be due to non-optimal storing conditions of the feed. Moreover, our results suggest that mycotoxin thresholds should be adapted to consider the frequent case of toxin co-occurrence. This approach would better reflect the real toxigenic risk of feedstuffs

Mapping and assessing ecosystem services for sustainable policy and decision-making in Eritrea

The mapping and assessment of ecosystems and their services (MAES) is key to inform sustainable policy and decision-making at national and sub-national levels. Responding to the paucity of research in sub-Saharan Africa, we conduct a pilot study for Eritrea that aims to map and assess the temporal dynamics of key ecosystems and their services. We reviewed policy and legal documents, analyzed land cover changes and estimated the potential for ecosystem services supply through an expert-based matrix approach. Our results showed that from 2015 to 2019, the potential supply of the ecosystem services analyzed (e.g., crop provisioning, water supply and recreation) increased, with the exception of wood supply. Overall, our study presents policy-relevant insights as to where to conserve, develop, or restore ecosystem services supply in Eritrea. Our approach is transferable to similar data scarce contexts and can thereby support policies toward more sustainable land development for people and nature

INTEGRATED APPROACH COMBINING DIRECT SURVEY AND OBSERVATION IN DAMAGE ANALYSIS IN CASE OF SOIL SETTLEMENT

Nowadays, different methods are used to study historical masonry buildings. Among these, for the study of architectures with complex geometry, the effectiveness of an integrated approach, that is a method of analysis combining different disciplines, is increasingly evident.The aim of this paper is to show the importance of combining direct observation with structural analysis in order to understand the level of safety in buildings with composite geometries.This paper describes the analysis executed in the XX century parish church of San Bernardino in Sesto Calende (Va), which displays serious cracks and damage caused by soil settlements.The integrated approach starts with historical analysis, by consulting all available documents and drawings. To understand the geometry of the structure a new survey has been made and a three-dimensional digital representation was modeled, by which better deriving the weight of all the elements in the construction, and to find the correct actions and thrust on arches, columns, bases and foundation. All this data was used in the structural analysis based on the static method of limit analysis. For the material behavior the model proposed by J. Heyman (1966) is used, considering the “no tension” failure criterion. The static theorem of minimum reactions for settled states enunciated by M. Como (2010) is employed for the analysis of soil settlement effects on the building’s response.The work shows how direct survey of geometry and damage of a complex building has an effective importance in the structural analysis to ensure Cultural Heritage preservation and safety.</p

Mapping and assessment of ecosystems and their services. Urban ecosystems

Action 5 of the EU Biodiversity Strategy to 2020 requires member states to Map and Assess the state of Ecosystems and their Services (MAES). This report provides guidance for mapping and assessment of urban ecosystems. The MAES urban pilot is a collaboration between the European Commission, the European Environment Agency, volunteering Member States and cities, and stakeholders. Its ultimate goal is to deliver a knowledge base for policy and management of urban ecosystems by analysing urban green infrastructure, condition of urban ecosystems and ecosystem services. This report presents guidance for mapping urban ecosystems and includes an indicator framework to assess the condition of urban ecosystems and urban ecosystem services. The scientific framework of mapping and assessment is designed to support in particular urban planning policy and policy on green infrastructure at urban, metropolitan and regional scales. The results are based on the following different sources of information: a literature survey of 54 scientific articles, an online-survey (on urban ecosystems, related policies and planning instruments and with participation of 42 cities), ten case studies (Portugal: Cascais, Oeiras, Lisbon; Italy: Padua, Trento, Rome; The Netherlands: Utrecht; Poland: Poznań; Spain: Barcelona; Norway: Oslo), and a two-day expert workshop. The case studies constituted the core of the MAES urban pilot. They provided real examples and applications of how mapping and assessment can be organized to support policy; on top, they provided the necessary expertise to select a set of final indicators for condition and ecosystem services. Urban ecosystems or cities are defined here as socio-ecological systems which are composed of green infrastructure and built infrastructure. Urban green infrastructure (GI) is understood in this report as the multi-functional network of urban green spaces situated within the boundary of the urban ecosystem. Urban green spaces are the structural components of urban GI. This study has shown that there is a large scope for urban ecosystem assessments. Firstly, urban policies increasingly use urban green infrastructure and nature-based solutions in their planning process. Secondly, an increasing amount of data at multiple spatial scales is becoming available to support these policies, to provide a baseline, and to compare or benchmark cities with respect to the extent and management of the urban ecosystem. Concrete examples are given on how to delineate urban ecosystems, how to choose an appropriate spatial scale, and how to map urban ecosystems based on a combination of national or European datasets (including Urban Atlas) and locally collected information (e.g., location of trees). Also examples of typologies for urban green spaces are presented. This report presents an indicator framework which is composed of indicators to assess for urban ecosystem condition and for urban ecosystem services. These are the result of a rigorous selection process and ensure consistent mapping and assessment across Europe. The MAES urban pilot will continue with work on the interface between research and policy. The framework presented in this report needs to be tested and validated across Europe, e.g. on its applicability at city scale, on how far the methodology for measuring ecosystem condition and ecosystem service delivery in urban areas can be used to assess urban green infrastructure and nature-based solutions

Determining the reference range of blood presepsin in term and preterm neonates

Introduction Sepsis is still a major cause of morbidity and mortality in neonates, especially in preterm infants. Mortality can reach 60-70% in very low birth weight infants (birthweight 0.20). This reduced model explains 3.8% of the total sum of squares. After adjustment for all the factors in the model, presepsin levels appear to be significantly lower in twins (496 pg/ml \uf0b1 65.5 vs 655 pg/ml \uf0b1 11.8) and in neonates with Apgar at 1 min 658 (644 pg/ml \uf0b1 11.8 vs 774 pg/ml \uf0b1 56.2). So none of the above factors seems worth to be taken into account in determining the reference limits for presepsin blood levels in healthy term neonates. Preterm neonates. The largest differences in presepsin level are observed between small for gestational age (SGA) (903 pg/ml \uf0b1 57.1) and adequate for gestational age (AGA) neonates (703 pg/ml \uf0b1 26.7), between neonates with and without mechanical ventilation at blood sampling (1090 pg/ml \uf0b1 86.9 vs 711 pg/ml \uf0b1 24.7) and at delivery (855 pg/ml \uf0b1 87.3 vs 729 pg/ml \uf0b1 25.8), between neonates with and without venous catether (801 pg/ml \uf0b1 47.5 vs 716 pg/ml \uf0b1 28.9), between neonates who underwent blood sampling after the 4th day or before (797 pg/ml \uf0b1 46.2 vs 716 pg/ml \uf0b1 29.2), between males and females (778 pg/ml \uf0b1 35.1 vs 701 pg/ml \uf0b1 34.7). All these factors, when simultaneously introduced into a multivariable linear model, explain only 18.8% of the total sum of squares. A second multivariable linear model was fitted after removing the factors that showed the lowest effect on presepsin level (those associated with a p-value >0.50). This reduced model explains 13.4% of the total sum of squares. A third and more parsimonious multivariable linear model was fitted after removing the factors that showed the lowest effect on presepsin level (those associated with a p-value >0.20). This reduced model explains 12.3% of the total sum of squares. After adjustment for all the factors in the model, presepsin levels result to be significantly lower in AGA neonates (706 pg/ml \uf0b1 25.7 vs 890 pg/ml \uf0b1 55.0) and between neonates with and without mechanical ventilation at blood sampling (1074 pg/ml \uf0b1 85.3 vs 712 pg/ml \uf0b1 24.2). Even in this case, none of the above factors is expected to substantially affect the reference limits for presepsin blood levels in preterm neonates. Conclusion Presepsin blood levels seem to be quite independent of most of maternal and neonatal conditions examined in this study both in preterm and term neonates. The factors exerting significant effects (multiple birth and Apgar at 1 min, in term neonates, weight by gestational age and mechanical ventilation in preterm neonates) are expected to affect presepsin reference limits only to minor extent. References [1] Evaluation of a newly identified soluble CD14 subtype as a marker for sepsis. Yaegashi Y., Shirakawa K., Sato N., Suzuki Y., Kojika M., Imai S., Takahashi G., Miyata M., Furusako S., Endo S. J Infect Chemother. 2005;11:234-8. [2] CDC/NHSN surveillance definition of health care-associated infection and criteria for specific types of infection in the acute care setting. Horan T.C., Andrus M., Dudeck M.A. Am J Infect Control. 2008;36:309-32

Reproducibility of the WHO histological criteria for the diagnosis of Philadelphia chromosome-negative myeloproliferative neoplasms.

This study, performed on behalf of the Italian Registry of Thrombocythaemias (Registro Italiano Trombocitemie), aimed to test the inter-observer reproducibility of the histological parameters proposed by the WHO classification for the diagnosis of the Philadelphia chromosome-negative myeloproliferative neoplasms. A series of 103 bone marrow biopsy samples of Philadelphia chromosome-negative myeloproliferative neoplasms consecutively collected in 2004 were classified according to the WHO criteria as follows: essential thrombocythaemia (n=34), primary myelofibrosis (n=44) and polycythaemia vera (n=25). Two independent groups of pathologists reviewed the bone marrow biopsies. The first group was asked to reach a collegial 'consensus' diagnosis. The second group reviewed individually all the cases to recognize the main morphological parameters indicated by the WHO classification and report their results in a database. They were subsequently instructed to individually build a 'personal' diagnosis of myeloproliferative neoplasms subtype just assembling the parameters collected in the database. Our results indicate that high levels of agreement ( 6570%) have been reached for about all of the morphological features. Moreover, among the 18 evaluated histological features, 11 resulted statistically more useful for the differential diagnosis among the different Philadelphia chromosome-negative myeloproliferative neoplasms. Finally, we found a high percentage of agreement (76%) between the 'personal' and 'consensus' diagnosis (Cohen's kappa statistic >0.40). In conclusion, our results support the use of the histological criteria proposed by the WHO classification for the Philadelphia chromosome-negative myeloproliferative neoplasms to ensure a more precise and early diagnosis for these patients