A conserved residue in the tip proteins of CS1 and CFA/I pili of enterotoxigenic Escherichia coli that is essential for adherence

Enterotoxigenic Escherichia coli associated with human diarrheal disease utilize any of a limited group of serologically distinguishable pili for attachment to intestinal cells. These include CS1 and CFA/I pili. We show here that chemical modification of arginyl residues in CS1 pili abolishes CS1-mediated agglutination of bovine erythrocytes, which serves as a model system for attachment. Alanine substitution of the single arginyl residue in CooA, the major pilin, had no effect on the assembly of pili or on hemagglutination. In contrast, substitution of alanine for R181 in CooD, the minor pilin associated with the pilus tip, abolished hemagglutination, and substitution of R20 reduced hemagglutination. Neither of these substitutions affected CS1 pilus assembly. This shows that CooD is essential for CS1-mediated attachment and identifies specific residues that are involved in receptor binding but not in pilus assembly. In addition to mediating agglutination of bovine erythrocytes, CFA/I also mediates agglutination of human erythrocytes. Substitution of R181 by alanine in the CooD homolog, CfaE, abolished both of these reactions. We conclude that the same region of the pilus tip protein is involved in adherence of CS1 and CFA/I pili, although their receptor specificities differ. This suggests that the region of the pilus tip adhesin protein that includes R181 might be an appropriate target for therapeutic intervention or for a vaccine to protect against human diarrhea caused by enterotoxigenic E. coli strains that have serologically different pili.

CooB plays a chaperone-like role for the proteins involved in formation of CS1 pili of enterotoxigenic Escherichia coli

CS1 pili serve as the prototype of a class of filamentous appendages found on the surface of strains of enterotoxigenic Escherichia coli. The four genes needed to synthesize functional CS1 pili in E. coli K12 are: cooA, which encodes the major pilin protein; cooD, which encodes a minor pilin protein found at the tip of the structure; cooC, which encodes a protein found in the outer membrane of piliated bacteria; and cooB. We show here that CooB, which is required for pilus assembly but is not part of the final structure, stabilizes CooA, CooC, and CooD. We previously reported that CooB is complexed with CooA in the periplasm and show here that CooB also is found complexed with CooD in the periplasm. CooB is associated with the membrane fraction only in the presence of CooC, suggesting that these two proteins also interact. This suggests that although it has no homology to known chaperone proteins, CooB serves a chaperone-like role for assembly of CS1.

Limiar anaerobio (4 mM de lactato sanguineo), velocidade critica determinada a partir de diferentes distancias e performance aerobia em nadadores e nadadoras de 10 a 15 anos

Universidade Estadual de Campinas . Faculdade de Educação Física

Aquisition and extinction of two characteristics consistent with fear relevance

Fear relevance, the potential of a stimulus to become quickly associated with fear, is a characteristic assumed to have an evolutionary basis and to result in preferential processing. Previous research has shown that fear relevant stimuli share a number of characteristics, negative valence and preferential identification in a visual search task, for instance. The present research examined whether these two characteristics can be acquired by non-fear relevant stimuli (geometric shapes) as a result of Pavlovian fear conditioning. Two experiments employed an aversive learning paradigm with geometric shape CSs and a shock US, with stimulus ratings, affective priming and visual search performance assessed before and after acquisition and after extinction. Differential electrodermal responses, larger during CS1 than CS, were present during acquisition but not during extinction. Affective priming results suggest that the CS1 acquired negative valence during acquisition, which was lost during extinction. However, negative valence as indexed by more negative ratings for CS1 than for CS shapes seemed to survive extinction. Preferential attentional processing as indexed by faster detection of CS1 among CS shapes than vice versa on the visual search task also remained. The current research confirmed that characteristics of fear relevant stimuli can be acquired in an aversive learning episode and that they may be extinguished. This supports the proposal that fear relevance may be malleable through learning.

Extinction of affective learning: No evidence for dual process accounts of human learning

The acquisition and extinction of affective valence to neutral geometrical shape conditional stimuli was investigated in three experiments. Experiment 1 employed a differential conditioning procedure with aversive shock USs. Differential electrodermal responding was evident during acquisition and lost during extinction. As indexed by verbal ratings, the CS1 acquired negative valence during acquisition,which was reduced after extinction. Affective priming, a reaction time based demand free measure of stimulus valence, failed to provide evidence for affective learning. Experiment 2 employed pictures of happy and angry faces as USs.Valence ratings after acquisitionweremore positive for theCS paired with happy faces (CS-H) and less positive for the CS paired with angry faces (CS-A) than during baseline. Extinction training reduced the extent of acquired valence significantly for both CSs, however, ratings of the CS-A remained different from baseline. Affective priming confirmed these results yielding differences between CS-A and CS-H after acquisition for pleasant and unpleasant targets, but for pleasant targets only after extinction. Experiment 3 replicated the design of Experiment 2, but presented the US pictures backwardly masked. Neither rating nor affective priming measures yielded any evidence for affective learning. The present results confirm across two different experimental procedures that, contrary to predictions from dual process accounts of human learning, affective learning is subject to extinction.

Siirtyminen C-kielestä Pythoniin ohjelmoinnin perusopetuksessa

Ohjelmointitaito on asia, jonka oppimisesta ja opettamisesta voidaan olla montaa mieltä, eikä yhtä oikeaa tapaa toteuttaa ohjelmoinnin opetusta tunnu olevan olemassa. Se on kuitenkin selvää, että jotkin menetelmät ja työkalut tuntuvat olevan parempia kuin toiset. Lukuvuoden 2005-2006 päätteeksi Lappeenrannan teknillinen yliopisto päätti päivittää ohjelmoinnin perusopetusta, ja kokeili siirtymistä Python-ohjelmointikieleen ohjelmoinnin alkeiskursseilla. Koska kurssin varsinaiset muutokset keskittyivät tekniseen infrastruktuuriin, tutustuttiin alustavassa kirjallisuustutkimuksessa ensin erilaisiin lähestymistapoihin,aiempiin tapauksiin sekä mielekkäiden työkalujen löytämiseen. Tässä diplomityössä perehdytään ohjelmoinnin opetuksen työkaluihin sekä erityisesti Python-ohjelmointikielen hyödyntämiseen ohjelmoinnin perusopetuksessa. Diplomityö esittelee useita lähestymistapoja sekä keskittyy tutkimaan Pythonin soveltuvuutta alkeisopetuksen käyttötarkoituksiin. Diplomityö tutustuu myös Lappeenrannassa järjestetyn ohjelmoinnin perusteiden kurssin tuloksiin, ja analysoi sitä, pystyikö Python-pohjainen kurssi toteuttamaan teknisen yliopiston sille asettamat vaatimukset. Lopuksi aineistosta analysoidaan jatkotutkimuksen tarpeita sekä pyritään löytämään ne osa-alueet, joita näissä jatkotutkimuksissa tulisi vielä kehittää.

Python as a programming language for the introductory programming courses

Tämä kandidaatintyö tutkii tietotekniikan perusopetuksessa keskeisen aiheen,ohjelmoinnin, alkeisopetusta ja siihen liittyviä ongelmia. Työssä perehdytään ohjelmoinnin perusopetusmenetelmiin ja opetuksen lähestymistapoihin, sekä ratkaisuihin, joilla opetusta voidaan tehostaa. Näitä ratkaisuja työssä ovat mm. ohjelmointikielen valinta, käytettävän kehitysympäristön löytäminen sekä kurssia tukevien opetusapuvälineiden etsiminen. Lisäksi kurssin läpivientiin liittyvien toimintojen, kuten harjoitusten ja mahdollisten viikkotehtävien valinta kuuluu osaksitätä työtä. Työ itsessään lähestyy aihetta tutkimalla Pythonin soveltuvuutta ohjelmoinnin alkeisopetukseen mm. vertailemalla sitä muihin olemassa oleviin yleisiin opetuskieliin, kuten C, C++ tai Java. Se tarkastelee kielen hyviä ja huonoja puolia, sekä tutkii, voidaanko Pythonia hyödyntää luontevasti pääasiallisena opetuskielenä. Lisäksi työ perehtyy siihen, mitä kaikkea kurssilla tulisi opettaa, sekä siihen, kuinka kurssin läpivienti olisi tehokkainta toteuttaa ja minkälaiset tekniset puitteet kurssin toteuttamista varten olisi järkevää valita.

Ohjelmoinnin perusteet -kurssin opintosuoritusten analyysi

The purpose of this Bachelor’s thesis is to review Fundamentals of programming course over six-year time period (2001–2006). We try to find out if students have passed the course in years 2005 and 2006, after the change of a lecturer, relatively more often than before. We also analyze the problem areas of the course so that these kinds of problems could be avoided in future. The work includes building of the database, creating of the statistics and analysis of the results. The statistics tell many things about development of the course but the analysis phase gives even more accurate information. In analysis phase we look for significant statistical differences with chi-square test and try to find causes and solutions for those differences. In this study we found out that the biggest proportion of passed students was in year 2006. However year 2005 had very bad results which were caused by many different factors. That’s why the null hypothesis, that year 2005 and 2006 had better results than the earlier years, has to be rejected. Despite that the good results in year 2006 imply that the course has developed in right direction.

Laajeneva ohjelmointiesimerkkikokoelma Ohjelmoinnin perusteet -kurssille

Lappeenrannan teknillisen yliopiston Ohjelmoinnin perusteet -kurssin osallistujilla on ollut vaikeuksia aloittelevan ohjelmoijan kannalta laajojen ohjelmien tuottamisessa ja tällaisten ohjelmien jäsentelemisessä aliohjelmiksi. Kurssin harjoitustyö on kuitenkin vaatinut tällaisen laajan ja jäsennellyn ohjelman tuottamista ja osallistujien ongelmat edellä mainittujen asioiden suhteen ovat ilmenneet palautetuista harjoitustöistä. Tässä työssä pyritään tuottamaan ratkaisu edellä kuvattuun ongelmaan konstruktiivisen tutkimuksen keinoin. Työssä tuotettava konstruktio on ohjelmointiesimerkkikokoelma ja sen avulla kurssin osallistujille voidaan esitellä kuinka laajoja ohjelmia voidaan tuottaa vaiheittain ja kuinka tällaisia ohjelmia voidaan jäsennellä aliohjelmiksi.

Harjoitustyön kehittäminen ohjelmoinnin perusteet kurssille

Työn tarkoituksena on luoda uusi harjoitustyö Ohjelmoinnin perusteet –kurssille. Harjoitustyön tavoitteena oli olla nykyistä työtä mielekkäämpi, sekä sidottu käytäntöön. Näiden avulla tavoiteltiin kurssin harjoitustyön suorittavien opiskelijoiden osuuden kasvattamista. Lopputuloksena on kolmeosainen harjoitustyö, jossa opiskelija suorittaa kuvamanipulointia sekä hyödyntää avoimia rajapintoja. Tulevaisuuteen jää haasteeksi harjoitustyön tarkastaminen ja siihen liittyvät ongelmat, sekä työn jatkokehitys.

β-carotene biotransformation to obtain aroma compounds

Carotenoids are important constituents of food due to their color and because their degradation products generate important volatile compounds in foods. Aroma compounds derived from carotenoids are widely distributed in nature, and they are precursors of many important aromas in foods such as fruits and in flowers as well. They present high aromatic potential and are therefore of great interest to the industries of aromas and fragrances. In this study, more than 300 previously isolated microorganisms with potential for biotransformation of β-carotene present in the culture medium were selected using the plate method; about 80 strains presented capacity to produce aroma compounds and 7 strains were selected by an untrained panel of tasters to generate aroma compounds. The β-ionone was the main compound produced by CS1 (34.0 mg.L-1) and CF9 (42.4 mg.L-1) microorganisms at 72 and 24 hours of fermentation, cultured with and without pre-inoculation, respectively. The β-damascone and pseudoionone were found in low concentrations, 1,1,6-trimethyl-1,2,3,4-tetrahydronaphthalen (TTN) was tentatively identified and other compounds such as apocarotenoids, apparently obtained from the cleavage of the central part of the carotenoid, were detected.

Commercial reviews in the tender process of contractors

Purpose – The purpose of this paper is to show the extent to which clients amend standard form contracts in practice, the locus of the amendments, and how contractors respond to the amendments when putting together a bid. Design/methodology/approach – Four live observational case studies were carried out in two of the top 20 UK construction firms. The whole process used to review the proposed terms and conditions of the contract was shadowed using participant observation, interview and documentary analysis. Findings – All four cases showed strong evidence of amendments relating mostly to payment and contractual aspects: 83 amendments in Case Study 1 (CS1), 80 in CS2, 15 in CS3 and 29 in CS4. This comprised clauses that were modified (37 per cent), substituted (23 per cent), deleted (7 per cent) and new additions (33 per cent). Risks inherent in the amendments were mostly addressed through contractual rather than price mechanisms, to reflect commercial imperatives. “Qualifications” and “clarifications” were included in the tender submissions for post-tender negotiations. Thus, the amendments did not necessarily influence price. There was no evidence of a “standard-form contract“ being used as such, although clients may draw on published “standard-form contracts” to derive the forms of contract actually used in practice. Practical implications – Contractors should pay attention to clauses relating to contractual and financial aspects when reviewing tender documents. Clients should draft equitable payment and contractual terms and conditions to reduce risk of dispute. Indeed, it is prudent for clients not to pass on inestimable risks. Originality/value – A better understanding of the extent and locus of amendments in standard form contracts, and how contractors respond, is provided.

Limiar anaeróbio e velocidade crítica determinada com diferentes distâncias em nadadores de 10 a 15 anos: relações com a performance e a resposta do lactato sanguíneo em testes de endurance

O objetivo deste estudo foi comparar a velocidade crítica (VC) determinada através de diferentes distâncias com o limiar anaeróbio (LAn) e as velocidades máximas mantidas em testes de 20 (V20) e 30 (V30) minutos na natação, verificando se a idade cronológica em jovens nadadores interfere nessas relações. Participaram do estudo 31 nadadores (17 meninas e 14 meninos) divididos segundo a idade cronológica em dois grupos: 10 a 12 anos e 13 a 15 anos. O LAn foi determinado como sendo a velocidade correspondente a 4mM de lactato sanguíneo. A VC1 (25/50/100m), VC2 (100/200/400m) e a VC3 (50/100/200m) foram calculadas através do coeficiente angular da reta de regressão linear entre as distâncias e seus respectivos tempos. As V20 e V30 foram determinadas através de três a seis repetições, com coletas de sangue no 10º minuto e ao final do tiro. Para o grupo de 10 a 12 anos, a VC1 (m/s) (0,98 ± 0,17) e o LAn (0,97 ± 0,12) não foram diferentes entre si, sendo maiores do que a VC2 (0,92 ± 0,16), VC3 (0,89 ± 0,18), V20 (0,92 ± 0,11) e V30 (0,90 ± 0,11). Para o grupo de 13 a 15 anos, a VC1 (m/s)(1,11 ± 0,11) foi maior do que o LAn (1,02 ± 0,07), V20 (0,99 ± 0,09), V30 (0,97 ± 0,09), VC2 (0,98 ± 0,11) e VC3 (1,00 ± 0,11). Pode-se concluir que a distância utilizada na determinação da VC interfere no valor obtido, independente da idade cronológica. A VC determinada com distâncias entre 50 e 400m pode ser utilizada na avaliação da capacidade aeróbia de crianças e adolescentes, substituindo os testes contínuos máximos com durações próximas a 20 ou 30 minutos.

Comparison between protocols for the determination of anaerobic threshold in adolescent swimmers

Aim. The purpose of this study was to compare the anaerobic threshold speed (AT) obtained from fixed lactate blood concentrations (AT 4 mM and AT 3.5 mM), lactate minimum speed (LM) and critical speed (CS), determined from different distances in fifteen Brazilian national level swimmers (10 boys = 14.8 ± 0.6 years old and 5 girls = 14.6 ±0.8 year-old). Methods. The tests to determine the AT 4 mM, AT 3.5 mM, LM and CS were performed in a 25 m swimming pool and consisted of 7 or 8 evaluations separated by 24-48 h intervals. Data were submitted to analysis of variance (ANOVA) for repeated measures, followed by the post hoc Scheffé test and Pearson correlation coefficients. Significance was set at P<0.01. Results. There were no significant differences among the values for AT 4 mM and CS1 (1.34 ± 0.05 vs. 1.33 ± 0.05 m.s -1, respectively). However, AT 4 mM and CS1 were significantly higher than AT 3.5 mM (1.28 ± 0.04 m.s -1), LM (1.27 ± 0.05 m.s -1), CS2 (1.26 ± 0.06 m.s -1), CS3 (1.27 ± 0.06 m.s -1) and CS4 (1.25 ± 0.07 m.s -1). There were no significant differences among the values for AT 3.5 mM, LM, CS2, CS3 and CS4. Conclusion. The results obtained in this study suggest that the anaerobic threshold determined by a fixed lactate concentration of 3.5 mM, as well as the LM and the CS methods determined by different distances, seem to be the most appropriate indexes for the evaluation of aerobic capacity in adolescent swimmers.