Genetic and antigenic analysis of Babesia bigemina isolates from five geographical regions of Brazil

A molecular epidemiological study was performed with Babesia bigemina isolates from five geographical regions of Brazil. The genetic analysis was done with random amplification of polymorphic DNA (RAPD), repetitive extragenic palindromic elements-polymerase chain reaction (REP-PCR) and enterobacterial repetitive intergenic consensus sequences-polymerase chain reaction (ERIC-PCR) that showed genetic polymorphism between these isolates and generated fingerprinting. In RAPD, ILO872 and ILO876 primers were able to detect at least one fingerprinting for each B. bigemina isolate. The amplification of B. bigemina DNA fragments by REP-PCR and ERIC-PCR gave evidence for the presence in this haemoprotozoan of the sequences described previously in microorganisms of the bacterial kingdom. For the first time it was demonstrated that both techniques can be used for genetic analysis of a protozoan parasite, although the ERIC-PCR was more discriminatory than REP-PCR. The dendogram with similarity coefficient among isolates showed two clusters and one subcluster. The Northeastern and Mid-Western isolates showed the greatest genetic diversity, while the Southeastern and Southern isolates were the closest. The antigenic analysis was done through indirect fluorescent antibody technique and Western blotting using a panel of monoclonal antibodies directed against epitopes on the merozoite membrane surface, rhoptries and membrane of infected erythrocytes. As expected, the merozoite variable surface antigens, major surface antigen (MSA)-1 and MSA-2 showed antigenic diversity. However, B cell epitopes on rhoptries and infected erythrocytes were conserved among all isolates studied. In this study it was possible to identify variable and conserved antigens, which had already been described as potential immunogens. Considering that an attenuated Babesia clone used as immunogen selected populations capable of evading the immunity induced by this vaccine, it is necessary to evaluate more deeply the cross-protection conferred by genetically more distant Brazilian B. bigemina isolates and make an evaluation of the polymorphism degree of variable antigens such as MSA-1 and MSA-2.

Expression and distribution of connexin 32 in rat liver with experimentally induced fibrosis

The connexin 32 (Cx32) is a protein that forms the channels that promote the gap junction intercellular communication (GJIC) in the liver, allowing the diffusion of small molecules through cytosol from cell-to-cell. Hepatic fibrosis is characterized by a disruption of normal tissue architeture by cellular lesions, and may alter the GJIC. This work aimed to study the expression and distribution of Cx32 in liver fibrosis induced by the oral administration of dimethylnitrosamine in female Wistar rats. The necropsy of the rats was carried out after five weeks of drug administration. They presented a hepatic fibrosis state. Sections from livers with fibrosis and from control livers were submitted to immunohistochemical, Real Time-PCR and Western-Blot analysis to Cx32. In fibrotic livers the Cxs were diffusely scattered in the cytoplasm, contrasting with the control livers, where the Cx32 formed junction plaques at the cell membrane. Also it was found a decrease in the gene expression of Cx32 without reduction in the protein quantity when compared with controls. These results suggest that there the mechanism of intercellular communication between hepatocytes was reduced by the fibrotic process, which may predispose to the occurrence of a neoplastic process, taken in account that connexins are considered tumor suppressing genes.

Development of IgY antibodies in chickens and IgG in rabbits immunized against proteins of Pythium insidiosum isolated from horses in the state of Rio de Janeiro

Pythiosis is caused by Pythium insidiosum and the occurrence of disease in horses was described in the North and Northwest State of Rio de Janeiro, Brazil. The disease was described in cattle, sheep, humans, and horses in different states and regions across the country. This paper describes the development of IgY and IgG polyclonal antibodies, in chicken and rabbits, respectively against proteins extracted from kunkers and hyphae of P. insidiosum from affected horses. The proteins were recognized by chicken, rabbit and horse antibodies by immunodiffusion and Western blot against majority bands of 27 and 43 KDa, and titrated by ELISA. The antibodies IgY developed by the first time against Brazilian strains of P. insidiosum may represent a valuable tool in the detection of antigens of the pathogen and contribute to further studies aimed at immunotherapy and knowledge about this disease in endemic areas in Rio de Janeiro and in Brazil.

Production and characterization of monoclonal antibodies against Campylobacter fetus subsp. venerealis

Myeloma cells Sp2/0-Ag14 and spleen cells from BALB/c mouse immunized with sonicated Campylobacter fetus subsp. venerealis NCTC 10354 were fused with polyethylene glycol (PEG) for the selection of clones producing antibodies. Clones were obtained by limiting dilution and screened for the production of specific antibodies to C. fetus subsp. venerealis NCTC 10354 by indirect ELISA and western blot against a panel of bacteria: C. fetus subsp. venerealis NCTC 10354, C. fetus subsp fetus ADRI 1812, C. sputorum biovar sputorum LMG 6647, C. lari NCTC 11352, and Arcobacter skirrowii LMG 6621 for the ELISA and C. fetus subsp. venerealis NCTC 10354 and C. sputorum biovar sputorum LMG 6647 for the western blotting. Fifteen clones producing monoclonal antibodies (MAbs) anti-C. fetus subsp. venerealis of the IgM (1) and IgG (14) classes were further screened for species-specificity. Four clones of the 15 obtained were producers of species-specific monoclonal antibodies (MAbs): two were specific for C. fetus subsp. venerealis and two were specific for C. fetus subsp. fetus. None of the clones were reactive against C. sputorum biovar sputorum LMG 6647. All clones recognized a protein with molecular mass of approximately 148 kDa from lysed C. fetus subsp. venerealis NCTC 10354.

Deregulation’s Impact on the Railway Freight Transport Sector’s Future in the Baltic Sea Region

The European transport market has confronted several changes during the last decade. Due to European Union legislative mandates, the railway freight market was deregulated in 2007. The market followed the trend started by other transport modes as well as other previously regulated industries such as banking, telecommunications and energy. Globally, the first country to deregulate the railway freight market was the United States, with the introduction of the Staggers Rail Act in 1980. Some European countries decided to follow suit already before regulation was mandated; among the forerunners were the United Kingdom, Sweden and Germany. The previous research has concentrated only on these countries, which has provided an interesting research gap for this thesis. The Baltic Sea Region consists of countries with different kinds of liberalization paths, including Sweden and Germany, which have been on the frontline, whereas Lithuania and Finland have only one active railway undertaking, the incumbent. The transport market of the European Union is facing further challenges in the near future, due to the Sulphur Directive, oil dependency and the changing structure of European rail networks. In order to improve the accessibility of this peripheral area, further action is required. This research focuses on topics such as the progression of deregulation, barriers to entry, country-specific features, cooperation and internationalization. Based on the research results, it can be stated that the Baltic Sea Region’s railway freight market is expected to change in the future. Further private railway undertakings are anticipated, and these would change the market structure. The realization of European Union’s plans to increase the improved rail network to cover the Baltic States is strongly hoped for, and railway freight market counterparts inside and among countries are starting to enhance their level of cooperation. The Baltic Sea Region countries have several special national characteristics which influence the market and should be taken into account when companies evaluate possible market entry actions. According to thesis interviews, the Swedish market has a strong level of cooperation in the form of an old-boy network, and is supported by a positive attitude of the incumbent towards the private railway undertakings. This has facilitated the entry process of newcomers, and currently the market has numerous operating railway undertakings. A contrary example was found from Poland, where the incumbent sent old rolling stock to the scrap yard rather than sell it to private railway undertakings. The importance of personal relations is highlighted in Russia, followed by the railway market’s strong political bond with politics. Nonetheless, some barriers to entry are shared by the Baltic Sea Region, the main ones being acquisition of rolling stock, bureaucracy and needed investments. The railway freight market is internationalizing, which is perceived via several alliances as well as the increased number of mergers and acquisitions. After deregulation, markets seem to increase the number of railway undertakings at a rather fast pace, but with the passage of time, the larger operators tend to acquire smaller ones. Therefore, it is expected that in a decade’s time, the number of railway undertakings will start to decrease in the deregulation pioneer countries, while the ones coming from behind might still experience an increase. The Russian market is expected to be totally liberalized, and further alliances between the Russian Railways and European railway undertakings are expected to occur. The Baltic Sea Region’s railway freight market is anticipated to improve, and, based on the interviewees’ comments, attract more cargoes from road to rail.

Analysis of corporate social responsibility (CSR) performance and financial performance of selected mining companies

The global concern about sustainability has been growing and the mining industry is questioned about its environmental and social performance. Corporate social responsibility (CSR) is an important issue for the extractive industries. The main objective of this study was to investigate the relationship between CSR performance and financial performance of selected mining companies. The study was conducted by identifying and comparing a selection of available CSR performance indicators with financial performance indicators. Based on the result of the study, the relationship between CSR performance and financial performance is unclear for the selected group of companies. The result is mixed and no industry specific realistic way to measure CSR performance uniformly is available. The result as a whole is contradictory and varies at company level as well as based on the selected indicators. The result of this study confirms that the relationship between CSR performance and financial performance is complicated and difficult to determine. As an outcome, evaluation of benefits of CSR in the mining sector could better be analyzed based on different attributes.

Perheyhtiö ja paikallisuus. A. Ahlström Osakeyhtiön historian perintö Noormarkussa

Tutkimus tarkastelee kulttuuriperinnön asemaa perheyrityksen yrityskulttuurin,identiteetin ja yrityskuvan osatekijänä ja sitä, miten historiatietoisuus ohjaa yrityksen toimintaa paikallisen vaikuttavuuden näkökulmasta. Samalla tutkimus selvittää, miten liiketoiminnan laajeneminen ja globalisoituminen vaikuttavat paikallistason toimintaan ja miten yritys käyttää kulttuuriperintöään muutosten hallinnassa. Tutkimus on historiallinen tapaustutkimus. Esimerkkitapauksena on A. Ahlström Osakeyhtiö ja Länsi-Suomessa sijaitseva Noormarkun ruukki, joka on ollut yrityksen omistuksessa 1870-luvulta lähtien. 1800-luvun lopusta 2000-luvulle ulottuva tutkimus perustuu pääasiassa arkistolähteisiin. Tutkimuksessa yhdistyvät kulttuuriperinnön ja liiketaloustieteen tutkimuksen sekä historiantutkimuksen lähestymistavat. Paikallistason toiminnan tarkastelussa tutkimus saa piirteitä myös tehdasyhdyskuntatutkimuksesta. Tutkimuksessa perheyrityksen kulttuuriperintöön sisällytetään aineellisen kulttuuriperinnön ja menneisyyden kuvausten lisäksi yrityksen jatkuvuuteen, omistajuuteen, kotipaikkaan ja yhteiskuntavastuuseen liittyvät perinteet. Tutkimus osoittaa, että historia ja perinteet voivat olla yritykselle voimavara mutta myös hidaste. Perheyhtiön traditio on kestänyt liiketoiminnallisista ja organisatorisista muutoksista huolimatta. Suvun rooli yhtiön sisällä on kuitenkin muuttunut. Perhejohtajuudesta on siirrytty sukuomistukseen. Ahlström on tyyppiesimerkki siitä, miten yritykset korostavat historian avulla toimintansa jatkuvuutta, luotettavuutta, uskottavuutta ja innovatiivisuutta. Yritys on rakentanut historiallista yrityskuvaansa muun muassa museoiden, näyttelyiden ja julkaisujen avulla. Yrityksen suhde historialliseen kotipaikkaansa, Noormarkkuun, on ollut poikkeuksellisen kiinteä. Noormarkun ruukki on esimerkki siitä, miten yrityksen menneisyyteen liittyvistä paikoista voi muodostua symbolisia muistin paikkoja. Historiatietoisuus synnyttää ja ylläpitää paikallisvastuuta, joka on ilmennyt eri aikakausina eri tavoin. Ruukin toimintoja on kehitetty, kun vanhat toiminnot ovat päättyneet. 2000-luvulla yritys hyödyntää kulttuuriperintöään liiketoiminnan resurssina, paikallisen liiketoiminnan ja matkailun kehittämisen lähtökohdista. Kansainvälisissä tutkimuksissa historian aktiivista käyttöä yrityksen strategisessa suunnittelussa kuvataan käsitteellä history management. Suomessa aihetta on toistaiseksi tutkittu vähän. Tutkimus on siten avaus yrityksiä kulttuuriperinnön säilyttäjinä ja käyttäjinä koskeville keskusteluille.

State of logistics and transport in the Baltic Sea region in 2025

The world’s pace of change is accelerating and new innovations, inventions and technologies come about every day. Change is unavoidable. It is difficult to keep up and even more difficult to prepare for the future. Even though it is not possible to know exactly what will happen in the future, by studying futures people can better anticipate what might lie ahead. By making decisions and realizing the consequences of their choices today, people and governments are able to actively decide how they will act in the future. Both opportunities and pitfalls lie ahead, which encourages actors to make more farsighted decisions. The Baltic Sea region is an interesting area for futures studies. It comprises 11 nations and more than 100 million inhabitants and entails countries with advanced, high-income economies, like Finland, Germany and Denmark, and developing economies, like Russia, Latvia and Lithuania. The western, eastern, northern and southern parts of the region are separated by the Baltic Sea, which at the same time represents a barrier and a facility for trade and travel between the countries belonging to the region The purpose of this study was to uncover the most probable future of transport and logistics in the Baltic Sea region in 2025 by using the Delphi method. Altogether 109 responses were collected in two separate instances from experts in all the Baltic Sea region countries, 56 of whom were defined as academic respondents and 53 of whom business respondents. Only minor differences in the opinions of academic and business experts were discovered, and the larger differences lie between eastern and western response groups. The Baltic Sea region is a very heterogeneous region and the division is clearest between East and West, which differ in political, economic, social, technological and environmental aspects. The probable future of the Baltic Sea region presented in this study is coherent with previous studies on the same subject. The future of the Baltic Sea region in terms of logistics and transport looks quite bright according to the experts who participated in the study. Trade volumes will grow and the importance of logistics and transport to the competitiveness of the region will increase. Respondents from eastern countries seemed to be more optimistic about the future in general. Most differences between opinions could be explained by the gap in technological and infrastructural development between the East and West. As eastern countries are less-developed in some parts of their economies, it is easier for them to improve the technical condition of infrastructure by merely catching up with the western countries.

Teollisen tuotannon ja palvelun ulkoistaminen Intiaan

Diplomityössä on tarkoituksena kartoittaa asioita, joita pitää ottaa huomioon pk-yrityksen tuotannon ulkoistamisessa Intiaan sekä laskemaan kuinka paljon halvemmaksi kiertovoitelukeskusten valmistaminen Intiassa tulee verrattuna Suomeen. Teoriaosuudessa on tarkasteltu Intian erilaista kulttuuria sekä kartoitettu asioita, jotka vaikuttavat länsimaalaisen yrityksen toimintaan Intiassa sekä mahdollisuuksia eri teollisuuden aloille. Empiriaosuudessa on keskitytty eri vaihtoehdoin kiertovoitelukeskuksen valmistuskustannuksiin ja niiden vertailuun Intian ja Suomen välillä. Tuloksena on voitu todeta, että valmistus Intiassa tulee selvästi halvemmaksi kuin Suomessa. Ensi askelten ottaminen luultavasti tulee olemaan vaikeaa lähinnä asiakkaiden löytämisessä ja verkostojen rakentamisessa. Riskejä oman tuotantolaitoksen perustamisessa Intiaan on, mutta myös mahdollisuudet ja potentiaali menestykselliseen liiketoimintaan ovat suuret. Ulkoistaminen on vain tehtävä maltillisesti ja järkevästi ja tehtyihin päätöksiin täytyy sitoutua.

International Advanced Water Technologies Centre, IAWTC/LADEC and LUT 2013 : Questionnaire Responses from Russian Drinking Water and Wastewater Treatment Plants

In this report, information is published concerning Russian water and wastewater treatment plants. The information is based on a questionnaire sent to 70 water and wastewater treatment plants in 2012-2013. The questionnaire was prepared by the International Advanced Water Technologies Centre (IAWTC) and Lahti Development Company (LADEC). The questions dealt with an assessment of the present state, the need for changes, renovation, investments, and how to improve the efficiency of the operation by training and investments. A significant need to renew the old pipelines, constructions, and processes was clearly evident. The aggregated answers can be utilized in Russia as internal benchmarking in order to arrange training and plant visits, which were requested in many of the answers. Sharing this open report with the respondents can aid networking and awareness of HELCOM requirements which relate to waste water treatment plants discharging their waste water directly or indirectly into the Baltic Sea. The aim of this report is to provide information for Finnish small and medium size companies (SMEs) as regards possible water related exportation to different parts of Russia.

The leaves of green plants as well as a cyanobacterium, a red alga, and fungi contain insulin-like antigens

We report the detection of insulin-like antigens in a large range of species utilizing a modified ELISA plate assay and Western blotting. We tested the leaves or aerial parts of species of Rhodophyta (red alga), Bryophyta (mosses), Psilophyta (whisk ferns), Lycopodophyta (club mosses), Sphenopsida (horsetails), gymnosperms, and angiosperms, including monocots and dicots. We also studied species of fungi and a cyanobacterium, Spirulina maxima. The wide distribution of insulin-like antigens, which in some cases present the same electrophoretic mobility as bovine insulin, together with results recently published by us on the amino acid sequence of an insulin isolated from the seed coat of jack bean (Canavalia ensiformis) and from the developing fruits of cowpea (Vigna unguiculata), suggests that pathways depending on this hormone have been conserved through evolution.

Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi

The aim of the present study was to demonstrate the presence of alpha-L-fucosidase in Trypanosoma cruzi. Immunocytochemical and biochemical techniques were used to localize and characterize a membrane-associated, neutral-pH-optimum, alpha-L-fucosidase from the parasite. Light and electron microscopy localized the alpha-L-fucosidase specifically on the surface of the parasite and on membranes in the posterior region of the epimastigote stage. Although much less intense, labeling was also detected on the surface of trypomastigotes. At least 50% of the alpha-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1% Triton X-100, suggesting that alpha-L-fucosidase is peripherally associated with membranes. The enzyme from epimastigotes had a neutral pH optimum (near 7) but displayed low specific activity when p-nitrophenyl-alpha-L-fucoside was employed as substrate (0.028 U/mg protein for epimastigotes and 0.015 U/mg protein for tissue culture-derived trypomastigotes). Polyacrylamide gel electrophoresis and Western blotting analysis both showed an expected 50-kDa polypeptide which was immunoreactive with anti-alpha-L-fucosidase antibodies.

Distortion and internal warping torsion of a double symmetric hollow section

This thesis is made in cooperation with Laboratory of Steel Structures and the steel company SSAB. Maximization of the benefits of high-strength steel usually requires the usage of thin wall thicknesses. This means the failures related to buckling, distortion and warping stand out. One must be aware of these phenomena to design thin-walled structures stressed with forces such as torsional loading. It is also important to take into account small stress ranges when evaluating the accurate fatigue strength of structures. The objective of this thesis is to clarify the theory of the uniform and non-uniform torsion. This paper focuses on warping due to the non-uniform torsion in double symmetric box girder and structural hollow section. The arisen stress states are explained and researched using the finite element method. Another research target is the distortion in double symmetric box girder due to torsion, and the restraining effect of transverse diaphragms at the load end. Multiple transverse diaphragms are used to study more efficient restraining against warping and distortion than a common one end plate structure.

Arsenic trioxide reduces the invasive and metastatic properties of nasopharyngeal carcinoma cells in vitro

Nasopharyngeal carcinoma (NPC) is notorious for the metastases, which are in close association with Epstein-Barr virus-encoded latent membrane protein 1 (LMP1). Arsenic trioxide (As2O3) has been shown to induce apoptosis and differentiation in NPC xenografts. Then, can it repress the cancer cells' metastasis potential? To elucidate this issue, the present study was performed. LMP1-negative cell line HNE1 and LMP1-positive cell line HNE1-LMP1 were used as in vitro model. Cells (1 x 10(5)/mL) were cultured with or without 3 µM As2O3 for 48 h. Then the survival cells were collected to investigate their potential of colony formation, attachment, invasion, and migration. Both confocal immunofluorescence staining and Western blot were used to detect the changes of LMP1 expression. The changes of MMP-9 were examined by RT-PCR assay and Western blot. The results were as follow: i) the colony formation inhibition rate (75.41 ± 3.9% in HNE1-LMP1 cells vs 37.89 ± 4.9% in HNE1 cells), the rate of attachment (HNE1-LMP1 vs HNE1: 56.40 ± 3.5 vs 65.87 ± 5.9%), the invasion inhibitory rate (HNE1-LMP1 vs HNE1: 56.50 ± 3.7 and 27.91 ± 2.1%), and the migration inhibitory rate (HNE1-LMP1 vs HNE1: 48.70 ± 3.9 vs 29.19 ± 6.27%) were all significantly different between the two cell lines (P < 0.01). ii) LMP1 was down-regulated in As2O3-treated HNE1-LMP1 cells. iii) The reduction of MMP-9 was found in As2O3-treated groups, more evident in HNE1-LMP1 cells. Thus, we conclude that As2O3 can reduce metastasis potential of NPC cells, involving inhibition of MMP-9 expression. LMP1 were also reduced in this process and seemed to enhance anti-metastasis activity of As2O3.

A DNA enzyme targeting Egr-1 inhibits rat vascular smooth muscle cell proliferation by down-regulation of cyclin D1 and TGF-β1

We have demonstrated that a synthetic DNA enzyme targeting early growth response factor-1 (Egr-1) can inhibit neointimal hyperplasia following vascular injury. However, the detailed mechanism of this inhibition is not known. Thus, the objective of the present study was to further investigate potential inhibitory mechanisms. Catalytic DNA (ED5) and scrambled control DNA enzyme (ED5SCR) were synthesized and transfected into primary cultures of rat vascular smooth muscle cells (VSMCs). VSMC proliferation and DNA synthesis were analyzed by the MTT method and BrdU staining, respectively. Egr-1, TGF-β1, p53, p21, Bax, and cyclin D1 expression was detected by RT-PCR and Western blot. Apoptosis and cell cycle assays were performed by FACS. Green fluorescence could be seen localized in the cytoplasm of 70.6 ± 1.52 and 72 ± 2.73% VSMCs 24 h after transfection of FITC-labeled ED5 and ED5SCR, respectively. We found that transfection with ED5 significantly inhibited cultured VSMC proliferation in vitro after 24, 48, and 72 h of serum stimulation, and also effectively decreased the uptake of BrdU by VSMC. ED5 specifically reduced serum-induced Egr-1 expression in VSMCs, further down-regulated the expression of cyclin D1 and TGF-β1, and arrested the cells at G0/G1, inhibiting entry into the S phase. FACS analysis indicated that there was no significant difference in the rate of apoptosis between ED5- and ED5SCR-transfected cells. Thus, ED5 can specifically inhibit Egr-1 expression, and probably inhibits VSMC proliferation by down-regulating the expressions of cyclin D1 and TGF-β1. However, ED5 has no effect on VSMC apoptosis.