Characterization of Xanthomonas axonopodis pv. phaseoli isolates

A simple, quick and easy protocol was standardized for extraction of total DNA of the bacteria Xanthomonas axonopodis pv. phaseoli. The DNA obtained by this method had high quality and the quantity was enough for the Random Amplified Polymorphic DNA (RAPD) reactions with random primers, and Polymerase Chain Reaction (PCR) with primers of the hypersensitivity and pathogenicity gene (hrp). The DNA obtained was free of contamination by proteins or carbohydrates. The ratio 260nm/380nm of the DNA extracted ranged from 1.7 to 1.8. The hrp gene cluster is required by bacterial plant pathogen to produce symptoms on susceptible hosts and hypersensitive reaction on resistant hosts. This gene has been found in different bacteria as well as in Xanthomonas campestris pv. vesicatoria (9). The primers RST21 and RST22 (9) were used to amplify the hrp gene of nine different isolates of Xanthomonas axonopodis pv. phaseoli from Botucatu, São Paulo State, Brazil, and one isolate, "Davis". PCR amplified products were obtained in all isolates pathogenic to beans.

Tool-supported invariant-based programming

The development of correct programs is a core problem in computer science. Although formal verification methods for establishing correctness with mathematical rigor are available, programmers often find these difficult to put into practice. One hurdle is deriving the loop invariants and proving that the code maintains them. So called correct-by-construction methods aim to alleviate this issue by integrating verification into the programming workflow. Invariant-based programming is a practical correct-by-construction method in which the programmer first establishes the invariant structure, and then incrementally extends the program in steps of adding code and proving after each addition that the code is consistent with the invariants. In this way, the program is kept internally consistent throughout its development, and the construction of the correctness arguments (proofs) becomes an integral part of the programming workflow. A characteristic of the approach is that programs are described as invariant diagrams, a graphical notation similar to the state charts familiar to programmers. Invariant-based programming is a new method that has not been evaluated in large scale studies yet. The most important prerequisite for feasibility on a larger scale is a high degree of automation. The goal of the Socos project has been to build tools to assist the construction and verification of programs using the method. This thesis describes the implementation and evaluation of a prototype tool in the context of the Socos project. The tool supports the drawing of the diagrams, automatic derivation and discharging of verification conditions, and interactive proofs. It is used to develop programs that are correct by construction. The tool consists of a diagrammatic environment connected to a verification condition generator and an existing state-of-the-art theorem prover. Its core is a semantics for translating diagrams into verification conditions, which are sent to the underlying theorem prover. We describe a concrete method for 1) deriving sufficient conditions for total correctness of an invariant diagram; 2) sending the conditions to the theorem prover for simplification; and 3) reporting the results of the simplification to the programmer in a way that is consistent with the invariantbased programming workflow and that allows errors in the program specification to be efficiently detected. The tool uses an efficient automatic proof strategy to prove as many conditions as possible automatically and lets the remaining conditions be proved interactively. The tool is based on the verification system PVS and i uses the SMT (Satisfiability Modulo Theories) solver Yices as a catch-all decision procedure. Conditions that were not discharged automatically may be proved interactively using the PVS proof assistant. The programming workflow is very similar to the process by which a mathematical theory is developed inside a computer supported theorem prover environment such as PVS. The programmer reduces a large verification problem with the aid of the tool into a set of smaller problems (lemmas), and he can substantially improve the degree of proof automation by developing specialized background theories and proof strategies to support the specification and verification of a specific class of programs. We demonstrate this workflow by describing in detail the construction of a verified sorting algorithm. Tool-supported verification often has little to no presence in computer science (CS) curricula. Furthermore, program verification is frequently introduced as an advanced and purely theoretical topic that is not connected to the workflow taught in the early and practically oriented programming courses. Our hypothesis is that verification could be introduced early in the CS education, and that verification tools could be used in the classroom to support the teaching of formal methods. A prototype of Socos has been used in a course at Åbo Akademi University targeted at first and second year undergraduate students. We evaluate the use of Socos in the course as part of a case study carried out in 2007.

IgA production, coliforms analysis and intestinal mucosa morphology of piglets that received probiotics with viable or inactivated cells

Two types of probiotics were used in piglets. One product is a mixed culture of viable Lactobacillus acidophilus, Enterococcus faecium e Bifidobacterium bifidum. The second product is composed of inactivated Lactobacillus acidophilus cells. The piglets received two weekly oral doses for 30 days while a control group did not receive probiotics. All piglets were euthanized at the 30th day of life and the mesenteric lymph nodes, the small intestine, and blood samples were collected. The tissue samples were studied by light microscopy and the blood serum was analyzed by ELISA method. The treatment with the probiotic with viable cells produced higher serum levels of IgA (P<0.05) and more IgA expressing cells were found in the mesenteric lymph nodes than observed in the inactivated cells treatment or control groups (P<0.05). Also, intestinal villi were longer, crypts were deeper (P<0.05) and fecal coliform count was lower than found in the inactivated product (P<0.05). These results suggest that viable probiotics are more efficient than inactivated probiotics to induce immunostimulation and intestinal modifications in piglets, thus improving their health and development.

Transfer of passive immunity and serum proteinogram in the first six months of life of Criollo Lageano and black and white holstein calves

The objective of this study was to evaluate and compare the transfer of passive immunity and the proteinogram in Criollo Lageano (CL) and Black and White Holstein (BWH) calves. Two groups were utilized with 13 Criollo Lageano and 10 BWH calves. Blood samples were collected for the measurement of total serum protein, electrophoresis of serum proteins, activity of the gamma glutamyl transferase, and concentration of IgG by the method of the zinc sulfate turbidity in periods between 24 and 36 hours of life, 15, 30, 60, 90, 120, 150 and 180 days. Statistical analysis was performed by ANOVA and Tukey test at 5% significance level, and correlations between variables were calculated. Variations of serum proteins followed a pattern of physiological behavior over the first six months of life and production of immunoglobulins was active earlier in BWH calves and slower in the Criollo Lageano, without causing any impact on their health. Gamma globulin in the first days of life (24-36h) was correlated with IgG (r=0.87 for CL and r=0.89 for BWH), PTS (r=0.91 for CL and r=0.92 for BWH), Glob (r=0.99 for CL and r=0.98 for BWH) and GGT (r=0.14 for CL and r=0.83 for BWH). It was concluded that there was no failure in the transfer of passive immunity in Criollo Lageano calves but this failure occurred in the BWH calves. IgG values estimated by the zinc sulfate turbidity and serum proteins were considered good indicators of the transfer of passive immunity in calves between 24 and 36 hours of life.

Toimittajan oikeus uutisoida ja toimitustyötä sääntelevät normit

Luettaessa uutisia lehdestä harvoin pohditaan sitä, onko toimituksellisessa työssä noudatettu lakia ja alan eettisiä ohjeita. Lukija luottaa siihen, että sääntöjä noudatetaan, mutta näin ei aina ole. Käytettäessä materiaalia toiselta lehdeltä jätetään alkuperäinen lähde usein mainitsematta. Tämän tutkielman tarkoituksena on selvittää keskeiset tekijänoikeuslain pykälät ja käytännössä kaikkein keskeisimmät Journalistin ohjeet sanomalehden toimittajan työssä ja miten nämä säännöt pääosin ilmenevät. Suoritetun kyselytutkimuksen perusteella tekijänoikeuslain pykälä oikeus uutisoida toisen lehden uutinen ja Journalistin ohjeista sääntö, jonka mukaan sisältöratkaisut on tehtävä journalistisin perustein, ovat keskeisimpiä sääntöjä käytännön työssä. Tutkimuksessa näitä sääntöjä käsitellään oikeusdogmaattisin menetelmin kieliopillisen tulkinnan kautta sekä toimittajan työn näkökulmasta. Tutkimuksen lainsäädännöllisen viitekehyksen muodostavat Suomen perustuslaki, laki sananvapauden käyttämisestä joukkoviestinnässä, kansallinen tekijänoikeuslaki ja Journalistin ohjeet. Tämän vuoksi tutkimuksessa on aiheellista tarkastella myös sananvapautta ja tekijänoikeuslakia toimituksellisessa työssä sekä näiden suhdetta toisiinsa ja myös median rakenteellista ja toiminnallista muutosta.

Magnetic properties of multi-walled nanotubes on polysterene substrate

Nanotubes are one of the most perspective materials in modern nanotechologies. It makes present investigation very actual. In this work magnetic properties of multi-walled nanotubes on polystyrene substrate are investigated by using quantum magnetometer SQUID. Main purpose was to obtain magnetic field and temperature dependences of magnetization and to compare them to existing theoretical models of magnetism in carbon-bases structures. During data analysis a mathematical algorithm for obtained data filtration was developed because measurement with quantum magnetometer assume big missives of number data, which contain accidental errors. Nature of errors is drift of SQUID signal, errors of different parts of measurement station. Nanotube samples on polystyrene substrate were studied with help of atomic force microscope. On the surface traces of nanotube were found contours, which were oriented in horizontal plane. This feature was caused by rolling method for samples. Detailed comparison of obtained dependences with information of other researches on this topic allows to obtain some conclusions about nature of magnetism in the samples. It emphasizes importance and actuality of this scientific work.

Käyttövarmuusanalyysin tuottama tärkeysjärjestys elintarviketeollisuuden prosessilaitteille – Suuntaviivat kustannustietoisen kunnossapitostrategian luomiselle

Hallitsematon ja reaktiivinen kunnossapito on eräs tuotannon suurimpia kustannustekijöistä. Suunnitelmallisesti ja systemaattisesti johdettuna kunnossapito on tuotantotehokkuuden suurin vaikuttaja. Merkittävä osa tuotannon tehokkuuden ylläpidosta saavutetaan laitteiden käyttövarmuudella. Käyttövarmuuden saaminen hallintaan perustuu ennakoivan kunnossapidon määrän kasvattamiseen. Samalla korjaavan kunnossapidon kustannusriski laskee ja siihen käytetty panos vähenee. Huonolla kunnossapidon suunnitelmallisuudella on päinvastaiset vaikutukset. Tavoitteena on määritellä prosessilaitteiden käyttövarmuuksiin perustuva laitekriittisyys. Tutkimuksessa yhdistetään riskien arviointimenetelmiä, joilla keskimääräiset vikavälit ja seuraukset valmistukseen mallinnetaan. Kriittisyystekijöitä ovat käytettävyys, luotettavuus, kustannustekijät, turvallisuus ja ympäristövaikutukset. Tekijöiden arvottamiseen kehitettiin riksianalyysitaulukko. Kriittisyysluokat jaettiin kolmeen kategoriaan, joista A on kriittisin, B keskinkertainen ja C on matalin luokka. Lähtötietojen keräys toteutettiin triangulaatiomenetelmää soveltaen. Empiirisessä osassa HKScan Oy:n lihanjalostustehtaan jauheliha- ja kestomakkaraosastojen laitteet jaettiin A-, B- ja C-luokkiin. Kriittisimpiä laitteita oli 20 prosenttia analysoidusta laitemäärästä. Nämä A-luokkaan sijoitetut laitteet aiheuttavat 80 prosenttia kustannusriskeistä. B-luokkaan kuuluu 50 prosenttia ja C-luokkaan 30 prosenttia laitteista. Luokittelusta erotettiin havaitut turvallisuusriskit riskienhallinnan toimenpiteitä varten. Kustannustietoinen kriittisyysluokittelu on pohja kunnossapitostrategian rakentamiselle. Tämän avuksi esitettiin taulukot huolto-ohjelman luomiseen ja luokituksien hyödyntämiseen päivittäisessä toiminnassa.

Fibrinolytic action on fresh human clots of whole body extracts and two semipurified fractions from Lonomia achelous caterpillar

The severe bleeding diathesis produced by intoxication with the venom of Lonomia achelous caterpillars is characterized by prolonged bleeding from superficial skin wounds as well as massive hemorrhage into body cavities. The aim of the present study was to evaluate the effect of the crude venom and its fibrinolytic fractions on in vitro lysis of whole blood clots. Venom fractions with fibrinolytic activity were obtained by gel filtration chromatography on Sephadex G75 using imidazole buffer, pH 7.4, at a flow rate of 24 ml/h. Four peaks with fibrinolytic activity were obtained by this method. The highest activity was found in the first two peaks (both peaks were used for the experiments). The results show that the caterpillar venom degraded the preformed clots at a slower rate than plasmin. In addition, plasma protease inhibitors of the fibrinolytic system (a2-antiplasmin, a2-macroglobulin, PAI, etc.) only weakly inhibited the lytic effect of the caterpillar venom. These characteristics, as well as the pattern of fibrinogen degradation products, the delay period on fibrin plate lysis and amidolytic activity on chromogenic substrate, reported previously, indicate that the caterpillar enzymes are different from plasmin and trypsin.

Changes in the TBARs content and superoxide dismutase, catalase and glutathione peroxidase activities in the lymphoid organs and skeletal muscles of adrenodemedullated rats

Thiobarbituric acid reactant substances (TBARs) content, and the activities of glucose-6-phosphate dehydrogenase (G6PDh), citrate synthase (CS), Cu/Zn- and Mn-superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPX) were measured in the lymphoid organs (thymus, spleen, and mesenteric lymph nodes (MLN)) and skeletal muscles (gastrocnemius and soleus) of adrenodemedullated (ADM) rats. The results were compared with those obtained for sham-operated rats. TBARs content was reduced by adrenodemedullation in the lymphoid organs (MLN (28%), thymus (40%) and spleen (42%)) and gastrocnemius muscle (67%). G6PDh activity was enhanced in the MLN (69%) and reduced in the spleen (28%) and soleus muscle (75%). CS activity was reduced in all tissues (MLN (75%), spleen (71%), gastrocnemius (61%) and soleus (43%)), except in the thymus which displayed an increment of 56%. Cu/Zn-SOD activity was increased in the MLN (126%), thymus (223%), spleen (80%) and gastrocnemius muscle (360%) and was reduced in the soleus muscle (31%). Mn-SOD activity was decreased in the MLN (67%) and spleen (26%) and increased in the thymus (142%), whereas catalase activity was reduced in the MLN (76%), thymus (54%) and soleus muscle (47%). It is particularly noteworthy that in ADM rats the activity of glutathione peroxidase was not detectable by the method used. These data are consistent with the possibility that epinephrine might play a role in the oxidative stress of the lymphoid organs. Whether this fact represents an important mechanism for the establishment of impaired immune function during stress remains to be elucidated.

Retinol-induced changes in the phosphorylation levels of histones and high mobility group proteins from Sertoli cells

Chromatin proteins play a role in the organization and functions of DNA. Covalent modifications of nuclear proteins modulate their interactions with DNA sequences and are probably one of the multiple factors involved in the process of switch on/off transcriptionally active regions of DNA. Histones and high mobility group proteins (HMG) are subject to many covalent modifications that may modulate their capacity to bind to DNA. We investigated the changes induced in the phosphorylation pattern of cultured Wistar rat Sertoli cell histones and high mobility group protein subfamilies exposed to 7 µM retinol for up to 48 h. In each experiment, 6 h before the end of the retinol treatment each culture flask received 370 KBq/ml [32P]-phosphate. The histone and HMGs were isolated as previously described [Moreira et al. Medical Science Research (1994) 22: 783-784]. The total protein obtained by either method was quantified and electrophoresed as described by Spiker [Analytical Biochemistry (1980) 108: 263-265]. The gels were stained with Coomassie brilliant blue R-250 and the stained bands were cut and dissolved in 0.5 ml 30% H2O2 at 60oC for 12 h. The vials were chilled and 5.0 ml scintillation liquid was added. The radioactivity in each vial was determined with a liquid scintillation counter. Retinol treatment significantly changed the pattern of each subfamily of histone and high mobility group proteins.

Increased fetal hemoglobin levels in patients infected with human immunodeficiency virus (HIV1/2)

Fetal hemoglobin was measured in HIV1/2 patients under treatment with combined therapy (zidovudine and a protease inhibitor). A total of 143 patients and 103 normal individuals were investigated by the quantitative method of Betke and the semi-quantitative acid elution method of Kleihauer. In the normal person, hemoglobin F makes up less than 1% and an increase higher than 1.5% was observed in 21.4% of HIV patients by the method of Betke and in 24.8% of HIV-infected patients by the method of Kleihauer. The quantitative biochemical method of Betke showed that the populations were significantly different (two-tailed Mann-Whitney test). The reason for this hemoglobin F increase might be ascribed to the effect of zidovudine or to direct viral action on gamma chain expression. The finding of a higher F cell frequency indicated by the method of Kleihauer rather suggests that there is an increased F cell clone proliferation rather than an increase in hemoglobin F level in every cell.

Epäsuoran kysyntätiedon hyödyntäminen verkkokaupan tuotevalikoima- ja hinnoittelupäätöksissä

Tässä diplomityössä tutkitaan, miten verkkokaupan kävijävirran käyttäytymistä analysoimalla voidaan tehdä perusteltuja, tarkoituksenmukaisiin nimikkeisiin ja niiden parametreihin kohdistuvia päätöksiä tilanteessa, jossa laajamittaisemmat historiatiedot toteutuneesta myynnistä puuttuvat. Teoriakatsauksen perusteella muodostettiin ratkaisumalli, joka perustuu potentiaalisten kysyntäajurien muodostamiseen ja testaamiseen. Testisarjan perusteella valittavaa ajuria käytetään estimoimaan nimikkeiden kysyntää, jolloin sitä voidaan käyttää toteutuneen myynnin sijasta esimerkiksi Pareto-analyysissä. Näin huomio on mahdollista keskittää rajattuun määrään merkitykseltään suuria nimikkeitä ja niiden yksityiskohtaisiin parametreihin, joilla on merkitystä asiakkaan ostopäätöstilanteissa. Lisäksi voidaan tunnistaa nimikkeitä, joiden ongelmana on joko huono verkkonäkyvyys tai yhteensopimattomuus asiakastarpeiden kanssa. Ajurien testaamisperiaatteena käytetään kertymäfunktioiden yhdenmukaisuustarkastelua, joka rakentuu kolmesta peräkkäisestä vaiheesta; visuaalisesta tarkastelusta, kahden otoksen 2-suuntaisesta Kolmogorov-Smirnov-yhteensopivuustestistä ja Pearsonin korrelaatiotestistä. Mallia ja sen avulla tuotettua kysynnän ajuria testattiin veneilyalan kuluttaja-asiakkaille suunnatussa verkkokaupassa, jossa sillä tunnistettiin Pareto-jakauman alkupäästä runsaasti nimikkeitä, joiden parametreissa oli myynnin kannalta epäedullisia tekijöitä. Jakauman toisessa päässä tunnistettiin satoja nimikkeitä, joiden ongelmana on ilmeisesti joko huono verkkonäkyvyys tai nimikkeiden yhteensopimattomuus asiakastarpeiden kanssa.

Variability of Vernier acuity measurements in untrained subjects of different ages

The Vernier acuity of 50 normal untrained subjects (20 males and 30 females) was measured by the method of adjustment. Subjects were divided into five age ranges with 10 subjects in each age group: 5-10, 11-20, 21-30, 31-40, and 41-50 years. All subjects had normal visual acuity (20/20) and volunteered to participate in the experiment. Children were selected from a local school and adults recruited from the technical staff of the Department of Ophthalmology of the School of Medicine. Vernier acuity was higher in adults compared to children. Intraindividual variability was high and it was estimated that for most individuals of all age groups a range of 100 to 700 trials was necessary to obtain a mean with a precision of 10%. These results suggest that Vernier acuity variability is an obstacle to its use in clinical settings.

Vedyn käytön tarkastelu Kemira Chemicals Oy:n Joutsenon tehtailla

Kemira Chemicals Oy:n Joutsenon tehtailla valmistetaan lipeää, suolahappoa, natriumhypokloriittia sekä natriumkloraattia. Lipeää, suolahappoa ja natriumhypokloriittia valmistetaan lipeätehtaassa. Natriumkloraattia valmistetaan kloraattitehtaassa. Kloraatti- ja lipeätehtaan tuotteet valmistetaan elektrolyysimenetelmällä. Elektrolyysien sivutuotteena syntyy vetykaasua, joka voidaan käyttää suolahapon valmistukseen, vetyvoimalaitoksen polttoaineena tai myydä asiakkaalle. Työn tavoitteena oli tarkastella vedyn käyttöä Joutsenon tehtailla. Tarkastelun tavoitteena oli löytää mahdollisia kehitys- tai jatkotutkimuskohteita vety- ja höyryjärjestelmästä. Koska vetyä käytetään myös vetyvoimalaitoksen polttoaineena, joka tuottaa tehtailla tarvittavan prosessihöyryn, tarkasteltiin työssä myös höyryn käyttöä tehtailla. Tarkastelua varten tehtiin Microsoft Excel-pohjainen taselaskentamalli, jolla simuloitiin vedyn ja höyryn käyttöä tehtailla. Työn tuloksena saatiin Excel-pohjainen simulointimalli, jolla pystyttiin tutkimaan vedyn ja höyryn käyttöä. Vedyn ja höyryn käyttöä tutkittiin viidessä eri skenaariossa. Skenaariossa yksi määritettiin pienimmät mahdolliset elektrolyysiin syötettävät sähkövirran arvot, joilla tehtaita on turvallista käyttää. Skenaariossa kaksi määritettiin pienimmät mahdolliset elektrolyysiin syötettävät sähkövirran arvot, joilla voimalaitoksen turbiini pysyisi ajossa. Skenaariossa kolme määritettiin tehtaiden tämän hetkinen maksimi kapasiteetti. Skenaarioissa neljä ja viisi tutkittiin, miten mahdollinen tehtaiden tuotantojen kasvattaminen vaikuttaisi vety- ja höyryjärjestelmään. Työn tuloksien perusteella kehitys- ja jatkotutkimuskohteita olisivat lipeän haihdutuksen höyryn kulutuksen pienentäminen, turbiinin käyttöajan kasvattaminen sekä eri lähteistä saatavan hukkalämmön parempi hyödyntäminen kaukolämmön tuotannossa. Tehtaiden tuotantoja kasvatettaessa on syytä kiinnittää huomioita myös voimalaitoksen pääkattilan ja turbiinin kapasiteettiin.

Toxicity of a cyanobacterial extract containing microcystins to mouse lungs

Toxic cyanobacteria in drinking water supplies can cause serious public health problems. In the present study we analyzed the time course of changes in lung histology in young and adult male Swiss mice injected intraperitoneally (ip) with a cyanobacterial extract containing the hepatotoxic microcystins. Microcystins are cyclical heptapeptides quantified by ELISA method. Ninety mice were divided into two groups. Group C received an injection of saline (300 µl, ip) and group Ci received a sublethal dose of microcystins (48.2 µg/kg, ip). Mice of the Ci group were further divided into young (4 weeks old) and adult (12 weeks old) animals. At 2 and 8 h and at 1, 2, 3, and 4 days after the injection of the toxic cyanobacterial extract, the mice were anesthetized and the trachea was occluded at end-expiration. The lungs were removed en bloc, fixed, sectioned, and stained with hematoxylin-eosin. The percentage of the area of alveolar collapse and the number of polymorphonuclear (PMN) and mononuclear cell infiltrations were determined by point counting. Alveolar collapse increased from C to all Ci groups (123 to 262%) independently of time, reaching a maximum value earlier in young than in adult animals. The amount of PMN cells increased with time of the lesion (52 to 161%). The inflammatory response also reached the highest level earlier in young than in adult mice. After 2 days, PMN levels remained unchanged in adult mice, while in young mice the maximum number was observed at day 1 and was similar at days 2, 3, and 4. We conclude that the toxins and/or other cyanobacterial compounds probably exert these effects by reaching the lung through the blood stream after ip injection.