Diagnosis of Leishmania infantum infection by Polymerase Chain Reaction in wild mammals

Visceral leishmaniasis is a chronic infectious disease caused by Leishmania infantum (synonym: Leishmania chagasi) and transmitted by the sandfly Lutzomyia longipalpis in Brazil. It is an endemic zoonosis in several regions of the country, including Belo Horizonte (State of Minas Gerais). In urban areas, the domestic dog is susceptible and considered the most important animal reservoir. However, L. infantum has been previously diagnosed in other species, including captive primates and canids. This study aimed to evaluate the presence of the agent DNA in captive animals as well as some free ranging animals from the Zoo-Botanical Foundation of Belo Horizonte by Polymerase Chain Reaction. Eighty one blood samples from primates, carnivores, ruminants, edentates, marsupial, and a monogastric herbivore were analyzed. Three primates Alouatta guariba (brown howler monkey), and two canids Speothos venaticus (bush dog) were positive, demonstrating the importance of leishmaniasis control in endemic areas for preservation of wildlife species in captivity.

Monitoring and molecular characterization of group Drotavirus in Brazilian poultry farms

Abstract: Rotaviruses are etiological agents of diarrhea both in humans and in several animal species. Data on avian Group D rotaviruses (RVD) are scarce, especially in Brazil. We detected RVD in 4 pools of intestinal contents of broilers, layer and broiler breeders out of a total of 111 pools from 8 Brazilian states, representing an occurrence of 3.6%, by a specific RVD RT-PCR targeting the VP6 gene. Phylogenetic tree confirmed that the Brazilian strains belong to group D and 3 of the sequences were identical in terms of amino acid whereas one showed 99.5% identity with the others. The sequences described in this study are similar to other sequences previously detected in Brazil, confirming the conserved nature of the VP6 protein.

Biomolecular screening for inhibitors of butyrylcholinesterase : identification and characterization using in vitro and in silico tools

Drug discovery is a continuous process where researchers are constantly trying to find new and better drugs for the treatment of various conditions. Alzheimer’s disease, a neurodegenerative disease mostly affecting the elderly, has a complex etiology with several possible drug targets. Some of these targets have been known for years while other new targets and theories have emerged more recently. Cholinesterase inhibitors are the major class of drugs currently used for the symptomatic treatment of Alzheimer’s disease. In the Alzheimer’s disease brain there is a deficit of acetylcholine and an impairment in signal transmission. Acetylcholinesterase has therefore been the main target as this is the main enzyme hydrolysing acetylcholine and ending neurotransmission. It is believed that by inhibiting acetylcholinesterase the cholinergic signalling can be enhanced and the cognitive symptoms that arise in Alzheimer’s disease can be improved. Butyrylcholinesterase, the second enzyme of the cholinesterase family, has more recently attracted interest among researchers. Its function is still not fully known, but it is believed to play a role in several diseases, one of them being Alzheimer’s disease. In this contribution the aim has primarily been to identify butyrylcholinesterase inhibitors to be used as drug molecules or molecular probes in the future. Both synthetic and natural compounds in diverse and targeted screening libraries have been used for this purpose. The active compounds have been further characterized regarding their potencies, cytotoxicity, and furthermore, in two of the publications, the inhibitors ability to also inhibit Aβ aggregation in an attempt to discover bifunctional compounds. Further, in silico methods were used to evaluate the binding position of the active compounds with the enzyme targets. Mostly to differentiate between the selectivity towards acetylcholinesterase and butyrylcholinesterase, but also to assess the structural features required for enzyme inhibition. We also evaluated the compounds, active and non-active, in chemical space using the web-based tool ChemGPS-NP to try and determine the relevant chemical space occupied by cholinesterase inhibitors. In this study, we have succeeded in finding potent butyrylcholinesterase inhibitors with a diverse set of structures, nine chemical classes in total. In addition, some of the compounds are bifunctional as they also inhibit Aβ aggregation. The data gathered from all publications regarding the chemical space occupied by butyrylcholinesterase inhibitors we believe will give an insight into the chemically active space occupied by this type of inhibitors and will hopefully facilitate future screening and result in an even deeper knowledge of butyrylcholinesterase inhibitors.

Additional revenue opportunities in pulp mills and their impacts on the kraft process

Although the concept of multi-products biorefinery provides an opportunity to meet the future demands for biofuels, biomaterials or chemicals, it is not assured that its implementation would improve the profitability of kraft pulp mills. The attractiveness will depend on several factors such as mill age and location, government incentives, economy of scale, end user requirements, and how much value can be added to the new products. In addition, the effective integration of alternative technologies is not straightforward and has to be carefully studied. In this work, detailed balances were performed to evaluate possible impacts that lignin removal, hemicelluloses recovery prior to pulping, torrefaction and pyrolysis of wood residues cause on the conventional mill operation. The development of mill balances was based on theoretical fundamentals, practical experience, literature review, personal communication with technology suppliers and analysis of mill process data. Hemicelluloses recovery through pre-hydrolysis of chips leads to impacts in several stages of the kraft process. Effects can be observed on the pulping process, wood consumption, black liquor properties and, inevitably, on the pulp quality. When lignin is removed from black liquor, it will affect mostly the chemical recovery operation and steam generation rate. Since mineral acid is used to precipitate the lignin, impacts on the mill chemical balance are also expected. A great advantage of processing the wood residues for additional income results from the fact that the pulping process, pulp quality and sales are not harmfully affected. For pulp mills interested in implementing the concept of multi-products biorefinery, this work has indicated possible impacts to be considered in a technical feasibility study.

The revival of international solidarity : the Internationale Arbeiterhilfe, Willi Münzenberg and the Comintern in Weimar Germany, 1921-1933

The aim of the study is to write the first comprehensive history of the Internationale Arbeiterhilfe (International Workers’ Relief) and its message of international solidarity during the Weimar Republic, 1921–1933. The Arbeiterhilfe was the Communist International’s (Comintern) primary international solidarity organisation of the time. The work is identified as a contribution to the transnational history of the interwar period as its main focus is not on governmental politics or intra-state relations, but is focused on the transnational world of an international organisation. The history of the Arbeiterhilfe provides the main springboard from which to write a contextually-based analysis of international solidarity during the Weimar Republic. The study highlights for the first time the importance of the German communist Willi Münzenberg (1889–1940), as the leader of the Arbeiterhilfe, in the history of international solidarity. The main question of this study is how an explicit use of language coupled with the visualisation and practices of solidarity were created through the Arbeiterhilfe. How was solidarity actually envisaged, organised and brought to life by the Arbeiterhilfe in Weimar Germany? How did its expressions of solidarity change over time? Throughout the thesis, the changing and complex character of solidarity is analysed. How was the Arbeiterhilfe’s message of solidarity created and changed in relation to the Comintern and the Soviet Union’s policies? How did the Arbeiterhilfe create a new culture of international solidarity thought film, cinema, illustrated newspapers and the organising of mass spectacles of international solidarity? The Arbeiterhilfe had its international headquarters in Berlin which functioned as the base, one could argue, for some of the inter-war period’s most spectacular solidarity campaigns. The Arbeiterhilfe constitutes a significant case study of an early international organisation as it was one of the first international organisations for global (albeit not universal) international solidarity which had unparalleled prospects to develop new transnational identifications and social ties. It could consequently be suggested that the Arbeiterhilfe in several ways could be perceived as a predecessor to several post-1945 transnational solidarity organisations and International Non-Governmental Organisations (INGOs).

Growth and yield of corn grain and green ear in competition with weeds

Studies on plant growth are interesting because they provide explanations for the factors that influence yield in various crops. The objective of this work was to evaluate growth and yield in corn cultivar AG1051, when in competition with weeds. Cultivar AG 1051 was submitted to two groups of treatments: weed control, and sampling periods for dry biomass evaluation. The weed control treatments consisted of hoeing (two hoeings performed at 20 and 40 days after sowing) and no hoeing. Sampling periods consisted of collecting the above-ground part and roots of corn every fifteen days, until 105 days after sowing (DAS); the first sampling was performed 30 DAS. A completely randomized block design with ten replicates was used. For the characteristics evaluated in a single season, statistical analyses were carried out as a random block experiment. For the characteristics evaluated in several periods, statistical analyses were carried out as random blocks with split-plots (weed control assigned to plots). Fourteen weed species, unevenly distributed throughout the experimental area, were the most important. The growth observed for the above-ground part and root system of corn was 30% smaller in the non-hoed plots, compared to the hoed plots. Lack of weed control increased dry matter of the above-ground part of the weeds and reduced the number of unhusked and husked marketable green ears by 23% and 49%, respectively. Grain yield reduction caused by lack of weed control reached 38%.

Excessive functions, Appell polynomials and optimal stopping

The main topic of the thesis is optimal stopping. This is treated in two research articles. In the first article we introduce a new approach to optimal stopping of general strong Markov processes. The approach is based on the representation of excessive functions as expected suprema. We present a variety of examples, in particular, the Novikov-Shiryaev problem for Lévy processes. In the second article on optimal stopping we focus on differentiability of excessive functions of diffusions and apply these results to study the validity of the principle of smooth fit. As an example we discuss optimal stopping of sticky Brownian motion. The third research article offers a survey like discussion on Appell polynomials. The crucial role of Appell polynomials in optimal stopping of Lévy processes was noticed by Novikov and Shiryaev. They described the optimal rule in a large class of problems via these polynomials. We exploit the probabilistic approach to Appell polynomials and show that many classical results are obtained with ease in this framework. In the fourth article we derive a new relationship between the generalized Bernoulli polynomials and the generalized Euler polynomials.

Crowdfunding and its utilization for startup finance in Finland – factors of a successful campaign

This thesis studies crowdfunding with qualitive methods to introduce the phenomenon as well as provide guidance to those interested in its utilization. Knowledge and ideas were gathered form several sources, from academic literature to commercial media and expert interviews. Crowdfunding has already demonstrated its ability to impact the startup scene but is still far from being utilized to its full extent, especially in Finland, where even its legality has been questioned. Crowd financing can provide capital to entrepreneurs who might not otherwise be able to obtain funding as well as enable crowdsourcing the funders in several ways. A successful campaign, however, requires a wealth of knowledge on the subject, careful planning and hard work on the implementation. The thesis will provide most benefit to entrepreneurs who are considering the use of this new form of finance, but should also be of value for investors, academics, politicians and everyone else interested in the subject.

Altmetrics in practice: How are institutional repositories using altmetrics today?

Workshop at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Integrating open articles with open data in the life sciences with Europe PubMed Central.

Presentation at Open Repositories 2014, Helsinki, Finland, June 9-13, 2014

Molecular Basis for Resistance to Fluazifop-P-Butyl in Itchgrass (Rottboellia cochinchinensis) from Costa Rica

Rottboellia cochinchinensis is an annual grass weed species known as itchgrass, or "caminadora" in America´s Spanish speaking countries, and has become a major and troublesome weed in several crops. The application of fluazifop-P-butyl at recommended rates (125 g a.i. ha-1) was observed to be failing to control itchgrass in a field in San José, Upala county, Alajuela province, Costa Rica. Plants from the putative resistant R. cochinchinensis population survived fluazifop-P-butyl when treated with 250 g a.i. ha-1 (2X label rate) at the three- to four-leaf stage under greenhouse conditions. PCR amplification and sequencing of partial carboxyl transferase domain (CT) of the acetyl-CoA carboxylase (ACCase) gene were used to determine the molecular mechanism of resistance. A single non-synonymous point mutation from TGG (susceptible plants) to TGC (putative resistant plants) that leads to a Trp-2027-Cys substitution was found. This Trp-2027-Cys mutation is known to confer resistance to all aryloxyphenoxyproprionate (APP) herbicides to which fluazifop-P-butyl belongs. To the best of our knowledge, this is the first report of fluazifop-P-butyl resistance and a mutation at position 2027 for a Costa Rican R. cochinchinensis population.

Regulation of HSF2 and its function in mitosis

The cell is continuously subjected to various forms of external and intrinsic proteindamaging stresses, including hyperthermia, pathophysiological states, as well as cell differentiation and proliferation. Proteindamaging stresses result in denaturation and improper folding of proteins, leading to the formation of toxic aggregates that are detrimental for various pathological conditions, including Alzheimer’s and Huntington’s diseases. In order to maintain protein homeostasis, cells have developed different cytoprotective mechanisms, one of which is the evolutionary well-conserved heat shock response. The heat shock response results in the expression of heat shock proteins (Hsps), which act as molecular chaperones that bind to misfolded proteins, facilitate their refolding and prevent the formation of protein aggregates. Stress-induced expression of Hsps is mediated by a family of transcription factors, the heat shock factors, HSFs. Of the four HSFs found in vertebrates, HSF1-4, HSF1 is the major stress-responsive factor that is required for the induction of the heat shock response. HSF2 cannot alone induce Hsps, but modulates the heat shock response by forming heterotrimers with HSF1. HSFs are not only involved in the heat shock response, but they have also been found to have a function in development, neurodegenerative disorders, cancer, and longevity. Therefore, insight into how HSFs are regulated is important for the understanding of both normal physiological and disease processes. The activity of HSF1 is mainly regulated by intricate post-translational modifications, whereas the activity of HSF2 is concentrationdependent. However, there is only limited understanding of how the abundance of HSF2 is regulated. This study describes two different means of how HSF2 levels are regulated. In the first study it was shown that microRNA miR-18, a member of the miR-17~92 cluster, directly regulates Hsf2 mRNA stability and thus protein levels. HSF2 has earlier been shown to play a profound role in the regulation of male germ cell maturation during the spermatogenesis. The effect on miR-18 on HSF2 was examined in vivo by transfecting intact seminiferous tubules, and it was found that inhibition of miR-18 resulted in increased HSF2 levels and modified expression of the HSF2 targets Ssty2 and Speer4a. HSF2 has earlier been reported to modulate the heat shock response by forming heterotrimers with HSF1. In the second study, it was shown that HSF2 is cleared off the Hsp70 promoter and degraded by the ubiquitinproteasome pathway upon acute stress. By silencing components of the anaphase promoting complex/cyclosome (APC/C), including the co-activators Cdc20 and Cdh1, it was shown that APC/C mediates the heatinduced ubiquitylation of HSF2. Furthermore, down-regulation of Cdc20 was shown to alter the expression of heat shock-responsive genes. Next, we studied if APC/C-Cdc20, which controls cell cycle progression, also regulates HSF2 during the cell cycle. We found that both HSF2 mRNA and protein levels decreased during mitosis in several but not all human cell lines, indicating that HSF2 has a function in mitotic cells. Interestingly, although transcription is globally repressed during mitosis, mainly due to the displacement of RNA polymerase II and transcription factors, including HSF1, from the mitotic chromatin, HSF2 is capable of binding DNA during mitosis. Thus, during mitosis the heat shock response is impaired, leaving mitotic cells vulnerable to proteotoxic stress. However, in HSF2-deficient mitotic cells the Hsp70 promoter is accessible to both HSF1 and RNA polymerase II, allowing for stress-inducible Hsp expression to occur. As a consequence HSF2-deficient mitotic cells have a survival advantage upon acute heat stress. The results, presented in this thesis contribute to the understanding of the regulatory mechanisms of HSF2 and its function in the heat shock response in both interphase and mitotic cells.

The genera Eriochrysis P. Beauv., Imperata Cirillo and Saccharum L. (Poaceae - Andropogoneae - Saccharinae) in the state of Rio Grande do Sul, Brazil

The subtribe Saccharinae belongs to the tribe Andropogoneae and comprises ca. 140 species in 13 genera, including Eriochrysis, Imperata and Saccharum. This work presents a survey of the species of these three genera in the state of Rio Grande do Sul. Intensive field collections were made in different physiographic regions of the state, as well as studies in several herbaria. The occurrence of three native species of each genus was confirmed in Rio Grande do Sul, in addition to sugar cane (Saccharum officinarum) that is cultivated in the region. Based on the material examined, Eriochrysis villosa is reported for the first time in the state of Paraná, Brazil. Identification keys for the species, descriptions, data on their geographical distributions, habitats and flowering and fruiting periods, as well as illustrations of important taxonomic characters are provided.

Corticolous cyanobacteria from tropical forest remnants in northwestern São Paulo State, Brazil

Cyanobacteria are common in aquatic environments but are also well-adapted to terrestrial habitats where they are represented by a diversified flora. The present study aimed to contribute to our taxonomic knowledge of terrestrial cyanobacteria by way of a floristic survey of the main components of corticolous communities found in seasonal semideciduous forest fragments. Samples of visible growths of Cyanobacteria, algae, and bryophytes found on tree bark were randomly collected and their taxonomies examined. Eighteen species of Cyanobacteria were found belonging to the genera Aphanothece, Chroococcus, Lyngbya, Phormidium, Porphyrosiphon, Hapalosiphon, Hassalia, Nostoc, Scytonema, and Stigonema. Many genera and species observed in the present work have been reported in previous surveys of the aerophytic flora in several regions of the world, although six species were described only on the basis of populations found in the forest fragments studied, which highlights the importance of taxonomic studies of cyanobacteria in these habitats.

Distribution of AMPA-type glutamate receptor subunits in the chick visual system

Several glutamate receptor (GluR) subunits have been characterized during the past few years. In the present study, subunit-specific antisera were used to determine the distribution of the AMPA-type glutamate receptor subunits GluR1-4 in retinorecipient areas of the chick brain. Six white leghorn chicks (Gallus gallus, 7-15 days old, unknown sex) were deeply anesthetized and perfused with 4% buffered paraformaldehyde and brain sections were stained using immunoperoxidase techniques. The AMPA-type glutamate receptor subunits GluR1, GluR2/3 and GluR4 were present in several retinorecipient areas, with varying degrees of colocalization. For example, perikarya in layers 2, 3, and 5 of the optic tectum contained GluR1, whereas GluR2/3 subunits appeared mainly in neurons of layer 13. The GluR4 subunit was only detected in a few cells of the tectal layer 13. GluR1 and GluR2/3 were observed in neurons of the nucleus geniculatus lateralis ventralis, whereas GluR4 was only present in its neuropil. Somata in the accessory optic nucleus appeared to contain GluR2/3 and GluR4, whereas GluR1 was the dominant subunit in the neuropil of this nucleus. These results suggest that different subpopulations of visual neurons might express different combinations of AMPA-type GluR subunits, which in turn might generate different synaptic responses to glutamate derived from retinal ganglion cell axons