Effects of thyroid hormones and aldosterone on mineralocorticoid binding sites in the toad bladder.

In the urinary bladder of the toad Bufo marinus triiodothyronine selectively inhibits the late effect of aldosterone on Na+ transport. We have investigated whether T3 might mediate its antimineralocorticoid action by controlling: i) the level of aldosterone binding sites in the soluble (cytosolic) pool isolated from tissues treated with T3 (60 nM) for up to 20 hr of incubation; ii) the kinetics of uptake of 3H-aldosterone into cytoplasmic and nuclear fractions after 2 or 20 hr of exposure to T3. The number and the affinity of Type I (high affinity, low capacity) and Type II (low affinity, high capacity) cytosolic binding sites (measured at 0 degrees C) did not vary significantly after 18 hr of exposure to T3, while aldosterone-dependent Na+ transport was significantly inhibited. In addition, T3 did not modify the kinetics of uptake (90 min) of 3H-aldosterone into cytoplasmic and nuclear fractions of toad bladder incubated in vitro at 25 degrees C. By contrast, aldosterone itself was able to down-regulate its cytosolic and nuclear binding sites after an 18-hr exposure to the steroid hormone (10 or 80 nM). T3 slightly (20%) but significantly potentiated the down regulation of nuclear binding sites. In conclusion, T3 does not appear to have major effects on the regulation of the aldosterone receptor, which could explain in a simple manner its antimineralocorticoid action.

The caspase 8 inhibitor c-FLIP(L) modulates T-cell receptor-induced proliferation but not activation-induced cell death of lymphocytes.

The caspase 8 inhibitor c-FLIP(L) can act in vitro as a molecular switch between cell death and growth signals transmitted by the death receptor Fas (CD95). To elucidate its function in vivo, transgenic mice were generated that overexpress c-FLIP(L) in the T-cell compartment (c-FLIP(L) Tg mice). As anticipated, FasL-induced apoptosis was inhibited in T cells from the c-FLIP(L) Tg mice. In contrast, activation-induced cell death of T cells in c-FLIP(L) Tg mice was unaffected, suggesting that this deletion process can proceed in the absence of active caspase 8. Accordingly, c-FLIP(L) Tg mice differed from Fas-deficient mice by showing no accumulation of B220(+) CD4(-) CD8(-) T cells. However, stimulation of T lymphocytes with suboptimal doses of anti-CD3 or antigen revealed increased proliferative responses in T cells from c-FLIP(L) Tg mice. Thus, a major role of c-FLIP(L) in vivo is the modulation of T-cell proliferation by decreasing the T-cell receptor signaling threshold.

Roman amphora trade across the Straits of Gibraltar: An ancient 'anti-economic practice'?

Summary. Olive oil and fish products from the south of Hispania and North Africa played an important role in the Roman economy. The authors call attention to the asymmetrical distribution of archaeological data available on this subject, in particular the location of amphora kilns, and try to give an explanation, based on the evolution of European archaeology in the twentieth century.

Looking for factors involved in the light-dependent degradation of phytochrome A

SUMMARY : Phytochromes constitute a family of red/far-red photoreceptors regulating all the major transitions during the life cycle of plants. In Arabidopsis, five members: phyA,_ B, C, D and E, were identified. Phytochromes are synthesized in their inactive red-light absorbing form called Pr. Upon light absorbance they convert to the far-red light absorbing Pfr form. The Pfr form is the active conformer which converts back to the Pr form either rapidly upon far-red perception or in a slower process called dark reversion. ph~A represents an exception, in that it does not significantly dark-revert and two specific processes have been developed by the plants to decrease the amount of biologically active phyA. The first one is alight-dependent repression of the PHYA gene expression and the second one is alight-dependent degradation of the phyA protein. The latter is the most efficient process to rapidly decrease the level of active phyA. The ability of plants to regulate the amount of active phyA is critical in a far-red rich environment, a situation observed under a canopy. In these conditions, phyA is essential to induce the germination and the deetiolation of the young seedling. Later in the development the ability of phyA to repress growth counteracts the shade avoidance response. Therefore decreasing the amount of phyA allows stem growth and to compete with neighbours for the light. In this thesis, I investigate the light-dependent degradation of phyA. I developed a reverse genetic approach based on the systematic analysis of the light-dependent accumulation of phyA in the different cullin mutant cull, cul3a; cul3b and cul4. This analysis allowed me to show that CUL1 and CUL3A-based E3 ligase complexes are involved in the regulation of phyA degradation. Surprisingly, our results also demonstrate that cu14 is not affected in the degradation of phyA whereas constitutive Photomorphogenic 1 (COP1) a subunit of one CUL4based E3 complex was reported to be involved. Further investigations showed that the phenotype of cop1 is conditional, the mutant being defective in phyA degradation only in the presence of metabolisable sugars. I also showed that phyA is degraded by a proteasome-dependent mechanism both in the cytoplasm and in the nucleus using mutants and transgenic lines affected in the localization of phyA. Interestingly, I observed that phyA degradation was faster in the nucleus than in the cytosol and that rapid degradation of Pr also occurred in the nucleus suggesting that cytosolic accumulation of phyA in the dark is a way to regulate its proteolysis. Finally, we identify a short region similar to a PEST sequence required for phyA stability and we developed a unbiased genetic screen to identify new components involved in the regulation of the light-dependent degradation of phyA. The significance of these results are discussed. RESUME : Les phytochromes (phy) constituent une famille de photorécepteurs absorbant la lumière rouge et rouge lointaine et régulant toutes les étapes de transitions majeures dans la vie des plantes. Chez Arabidopsis, cinq membres : phyA, B, C, D et E ont été identifiés. Les phytochromes sont synthétisés sous une forme inactive appelée Pr absorbant la lumière rouge. Après perception de lumière ils passent sous une forme active Pfr absorbant dans le rouge lointain. La forme Pfr peut retourner sous la forme Pr après absorption de lumiëre rouge lointaine ou dans un processus lent appelé «réversion à l'obscurité ». phyA représente une exception à cette règle car il ne retoune pas significativement sous sa forme inactive dans le noir. Deux processus spécifiques ont donc été développés pour diminuer le taux de phyA actif. Le premier consiste en la répression du gène PHYA en condition de lumière et le second en une dégradation induite par la lumière de la protéine phyA. Ce dernier processus est le plus efficace pour diminuer rapidement le niveau de phyA. La capacité des plantes à réguler le taux de phyA actifs est critique dans un environnement riche en lumière rouge lointaine, une situation observée sous une canopée. Sous une canopée, phyA est essentiel pour induire la germination et la dé-étiolation de la jeune pousse. Plus tard dans le développement la capacité de phyA de réprimer la croissance freine la «réponse à l'évitement de l'ombre ». Par conséquent diminuer le taux de phyA permet la croissance de la tige et donc de rentrer en compétition pour la lumière avec les plantes avoisinantes. Dans cette thèse, j'ai étudié la dégradation de phyA. J'ai développé une approche génétique inverse basée sur l'analyse systématique de l'accumulation de phyA en condition de lumière dans les différents mutants cullin, cul1, cul3a, cul3b et cul4. Ces analyses nous ont permis d'identifier qu'un complexe E3 ligase CUL1 et un complexe E3 ligase CUL3A sont impliqués dans la régulation de la dégradation de phyA. Mes résultats démontrent aussi que le mutant cul4 n'est pas affecté dans la dégradation de phyA alors que Çonstitutive Photomorphogenic 1 (COPI) une sous unité d'un complexe CUL4 à été identifier dans la régulation de cette dégradation. Des analyses supplémentaires suggèrent que l'effet de la mutation cop1 est dépendante dë la présence de sucres métabolisables. J'ai aussi montré que phyA est dégradé dans le noyau et dans le cytoplasme par un mécanisme dépendant du protéasome et que la dégradation dans le.noyau est non seulement aspécifique de la forme Pr ou Pfr mais aussi est plus rapide que dans le cytoplasme. Ceci suggère que l'accumulation de phyA dans le cytoplasme permet son accumulation à des niveaux élevés à l'obscurité. Enfin j'ai identifié une région similaire à un motif PEST requise pour la stabilité de phyA et j'ai aussi développé un criblage génétique non biaisé pour identifier de nouveaux composants impliqués dans la régulation de la dégradation de phyA. L'importance de ces résultats est discutée dans le dernier chapitre de cette thèse.

Regulation by aldosterone of Na+,K+-ATPase mRNAs, protein synthesis, and sodium transport in cultured kidney cells.

Transepithelial Na+ reabsorption across tight epithelia is regulated by aldosterone. Mineralocorticoids modulate the expression of a number of proteins. Na+,K+-ATPase has been identified as an aldosterone-induced protein (Geering, K., M. Girardet, C. Bron, J. P. Kraehenbuhl, and B. C. Rossier, 1982, J. Biol. Chem., 257:10338-10343). Using A6 cells (kidney of Xenopus laevis) grown on filters we demonstrated by Northern blot analysis that the induction of Na+,K+-ATPase was mainly mediated by a two- to fourfold accumulation of both alpha- and beta-subunit mRNAs. The specific competitor spironolactone decreased basal Na+ transport, Na+,K+-ATPase mRNA, and the relative rate of protein biosynthesis, and it blocked the response to aldosterone. Cycloheximide inhibited the aldosterone-dependent sodium transport but did not significantly affect the cytoplasmic accumulation of Na+,K+-ATPase mRNA induced by aldosterone.

Receptor occupancy vs. induction of Na+-K+-ATPase and Na+ transport by aldosterone.

In the urinary bladder of the toad Bufo marinus aldosterone (between 0.8 and 100 nM) stimulates Na+ transport [half-maximal induction concentration (K1/2) = 6.5 nM]. At low hormone concentrations (0.8-8 nM), the increase of Na+ transport between 0.75 and 2.5 h is accompanied by a fall in transepithelial resistance (R). Higher hormone concentrations (30-800 nM) induce an additional resistance-independent fraction of Na+ transport within 2.5-8 h. From 6 h on, aldosterone (between 0.2 and 20 nM) stimulates in the same tissue the biosynthesis rate of the alpha- and beta-subunits of Na+-K+-ATPase (K1/2 = 3 and 1.5 nM, respectively). New pump synthesis is thus not a prerequisite for the early mineralocorticoid response but might be linked to the late transport event. The mineralocorticoid response is usually ascribed to interaction with the higher affinity type 1 receptor. In the present study we show, however, that at least 55% of the overall Na+ transport response is linked to nuclear occupation of the lower affinity type 2 receptors [dissociation constant (Kd) = 50 nM, maximum number of binding sites (Nmax) = 315 fmol/mg protein]. Distinct aldosterone effects, such as the fall in R and the increase in Na+-K+-ATPase synthesis, are more closely related to occupation of type 1 receptors (Kd = 0.3 nM, Nmax = 23 fmol/mg protein). At maximal induction of these latter parameters, only about 20% of type 2 receptors are occupied. These results suggest that both types of aldosterone receptors are involved in the mediation of the full mineralocorticoid response: type 1 in the early and late and type 2 particularly in the late tissue response.

ERK1/2 controls Na,K-ATPase activity and transepithelial sodium transport in the principal cell of the cortical collecting duct of the mouse kidney.

The collecting duct of normal kidney exhibits significant activity of the MEK1/2-ERK1/2 pathway as shown in vivo by immunostaining of phosphorylated active ERK1/2 (pERK1/2). The MEK1/2-ERK1/2 pathway controls many different ion transports both in proximal and distal nephron, raising the question of whether this pathway is involved in the basal and/or hormone-dependent transepithelial sodium reabsorption in the principal cell of the cortical collecting duct (CCD), a process mediated by the apical epithelial sodium channel and the basolateral sodium pump (Na,K-ATPase). To answer this question we used ex vivo microdissected CCDs from normal mouse kidney or in vitro cultured mpkCCDcl4 principal cells. Significant basal levels of pERK1/2 were observed ex vivo and in vitro. Aldosterone and vasopressin, known to up-regulate sodium reabsorption in CCDs, did not change ERK1/2 activity either ex vivo or in vitro. Basal and aldosterone- or vasopressin-stimulated sodium transport was down-regulated by the MEK1/2 inhibitor PD98059, in parallel with a decrease in pERK1/2 in vitro. The activity of Na,K-ATPase but not that of epithelial sodium channel was inhibited by MEK1/2 inhibitors in both unstimulated and aldosterone- or vasopressin-stimulated CCDs in vitro. Cell surface biotinylation showed that intrinsic activity rather than cell surface expression of Na,K-ATPase was controlled by pERK1/2. PD98059 also significantly inhibited the activity of Na,K-ATPase ex vivo. Our data demonstrate that the ERK1/2 pathway controls Na,K-ATPase activity and transepithelial sodium transport in the principal cell and indicate that basal constitutive activity of the ERK1/2 pathway is a critical component of this control.

Proteomic and transcriptomic profiling of Staphylococcus aureus surface LPXTG-proteins: correlation with agr genotypes and adherence phenotypes.

Staphylococcus aureus infections involve numerous adhesins and toxins, which expression depends on complex regulatory networks. Adhesins include a family of surface proteins covalently attached to the peptidoglycan via a conserved LPXTG motif. Here we determined the protein and mRNA expression of LPXTG-proteins of S. aureus Newman in time-course experiments, and their relation to fibrinogen adherence in vitro. Experiments were performed with mutants in the global accessory-gene regulator (agr), surface protein A (Spa), and fibrinogen-binding protein A (ClfA), as well as during growth in iron-rich or iron-poor media. Surface proteins were recovered by trypsin-shaving of live bacteria. Released peptides were analyzed by liquid chromatography coupled to tandem mass-spectrometry. To unambiguously identify peptides unique to LPXTG-proteins, the analytical conditions were refined using a reference library of S. aureus LPXTG-proteins heterogeneously expressed in surrogate Lactococcus lactis. Transcriptomes were determined by microarrays. Sixteen of the 18 LPXTG-proteins present in S. aureus Newman were detected by proteomics. Nine LPXTG-proteins showed a bell-shape agr-like expression that was abrogated in agr-negative mutants including Spa, fibronectin-binding protein A (FnBPA), ClfA, iron-binding IsdA, and IsdB, immunomodulator SasH, functionally uncharacterized SasD, biofilm-related SasG and methicillin resistance-related FmtB. However, only Spa and SasH modified their proteomic and mRNA profiles in parallel in the parent and its agr- mutant, whereas all other LPXTG-proteins modified their proteomic profiles independently of their mRNA. Moreover, ClfA became highly transcribed and active in fibrinogen-adherence tests during late growth (24 h), whereas it remained poorly detected by proteomics. On the other hand, iron-regulated IsdA-B-C increased their protein expression by >10-times in iron-poor conditions. Thus, proteomic, transcriptomic, and adherence-phenotype demonstrated differential profiles in S. aureus. Moreover, trypsin peptide signatures suggested differential protein domain exposures in various environments, which might be relevant for anti-adhesin vaccines. A comprehensive understanding of the S. aureus physiology should integrate all three approaches.

Hormonal regulation of (Na+,K+)-ATPase biosynthesis in the toad bladder. Effect of aldosterone and 3,5,3'-triiodo-L-thyronine.

Aldosterone stimulates transepithelial Na+ transport in the toad bladder, and thyroid hormone antagonizes this mineralocorticoid action. In the present study, we assessed the influence of these two hormones on the biosynthesis of (Na+,K+)ATPase, the major driving force of Na+ transport. Rates of enzyme synthesis were estimated by immunoprecipitation with monospecific alpha (96,000 daltons) and beta (60,000 daltons) subunit antibodies. After a 30-min pulse of intact tissue with [35S]methionine, the anti-alpha-serum recognized the 96,000-dalton alpha subunit and the anti-beta-serum, a 42,000-dalton protein, in total cell extracts. The biosynthesis rates of both these proteins were increased 2.8- and 2.4-fold respectively, over controls by 80 nM aldosterone after 18 h of hormone treatment. The hormonal effect was not apparent up to 3 h of incubation and was dose dependent between 0.2 and 20 nM aldosterone. The hormonal induction was antagonized by spironolactone (500-fold excess) but not by amiloride. The action of aldosterone thus seems to be a receptor-mediated process and a primary event independent of the Na+ permeability of the apical membrane. Thyroid hormone, on the other hand, had no effect on either basal or aldosterone-stimulated synthesis rates of both enzyme proteins. The results demonstrate a direct effect of aldosterone on gene expression of the (Na+,K+)-ATPase. Ultimately, this phenomenon could be linked to the late mineralocorticoid action of this hormone. On the other hand, thyroid hormone, in contrast to the situation in mammals, does not stimulate de novo enzyme synthesis in amphibia. Neither can the antimineralocorticoid action of thyroid hormone in the toad bladder be explained by an inhibition of the (Na+,K+)-ATPase synthesis.

Rakennukset ja rakentajat Raision Ihalassa rautakauden lopulla ja varhaisella keskiajalla

Turun yliopiston arkeologian oppiaine tutki Raision Ihalan historiallisella kylätontilla, ns. Mullin eduspellolla, asuinpaikan, josta löydettiin maamme oloissa harvinaisen hyvin säilyneitä rakennusten puuosien jäännöksiä. Löytö on ainutlaatuinen Suomen oloissa ja sillä on kansainvälistäkin merkitystä, koska hyvin säilyneet myöhemmän rautakauden ja varhaisen keskiajan maaseutuasuinpaikat, joista tavataan puujäännöksiä, ovat harvinaisia erityisesti itäisen Itämeren piirissä. Rakennukset on ennallistettu käyttäen tiukkaa paikallisen analogian (’Tight Local Analogy’) metodia, erityisesti suoraa historiallista analogista lähestymistapaa. Tätä tarkoitusta varten muodostettiin aluksi arkeologinen, historiallinen ja etnografinen lähdemalli. Tämä valittiin maantieteellisesti ja ajallisesti relevantista tutkimusaineistosta pohjoisen Itämeren piiristä. Tiedot lounaisen Suomen rakennuksista ja rakennusteknologiasta katsottiin olevan tärkein osa mallia johtuen historiallisesta ja spatiaalisesta jatkuvuudesta. Lähdemalli yhdistettiin sitten Mullin arkeologiseen aineistoon ja analyysin tuloksena saatiin rakennusten ennallistukset. Mullista on voitu ennallistaa ainakin kuusi eri rakennusta neljässä eri rakennuspaikassa. Rakennusteknologia perustui kattoa kannattaviin horisontaalisiin pitkiin seinähirsiin, jotka oli nurkissa yhdistetty joko salvoksella tai varhopatsaalla. Kaikissa rakennuksissa ulkoseinän pituus oli 5 – 7 metriä. Löydettiin lisäksi savi- ja puulattioita sekä kaksi tulisijaa, savikupoliuuni ja avoin liesi. Runsaan palaneen saven perusteella on mahdollista päätellä, että katto oli mitä todennäköisimmin kaksilappeinen vuoliaiskatto, joka oli katettu puulla ja/tai turpeella. Kaikki rakennukset olivat samaa tyyppiä ja ne käsittivät isomman huoneen ja kapean eteisen. Kaikki analysoitu puu oli mäntyä. Ulkoalueelta tavattiin lisäksi tunkioita, ojia, aitoja ja erilaisia varastokuoppia. Rakennukset on ajoitettu 900-luvun lopulta 1200-luvun lopulle (cal AD). Lopuksi tutkittiin rakennuksia yhteisöllisessä ympäristössään, niiden ajallista asemaa sekä asukkaiden erilaisia spatiaalisia kokemuksia ja yhteyksiä. Raision Ihalaa analysoidaan sosiaalisen identiteetin ja sen materiaalisten ilmenemismuotojen kautta. Nämä sosiaaliset identiteetit muodostuvat kommunikaatioverkostoista eri spatiaalisilla ja yhteisöllisillä ta¬soilla. Näitä eri tasoja ovat: 1) kotitalous arjen toimintoineen, perhe ja sukulaisuussuhteet traditioineen; 2) paikallinen identiteetti, rakennus, rakennuspaikka, asuinpaikan ympäristö ja sen käyttö, (maa)talo ja kylä; 3) Raision Ihalan kylä laajemmassa alueellisessa kontekstissaan pohjoisen Itämeren piirissä: kauppiaiden ja käsityöläisten kontaktiverkostot, uskonnollinen identiteetti ja sen muutokset.

Incorporação de dióxido de titânio em zeólitas para emprego em fotocatálise heterogênea

This work proposes the study of heterogeneous photocatalysis using TiO2 impregnated in zeolites beta, ZSM-5, mordenite, NaXb, NaXp and NaY for the decomposition of methylene blue. The catalysts were characterized by XRD, IR, textural analyses by N2 adsorption, SEM, DRS and the reaction of decomposition was monitored by UV visible. The results indicated that didn't have structural changes in the catalysts after Ti impregnations, only in the case of NaY and NaX zeolites. The better photocatalyst to metylene blue decomposition was beta/Ti zeolite due had one structure more accessible (with bigger porous) helping in TiO2 dispersion and catalytic active.

Caedes C. J. Caesari injuste illata

Invocatio: B.C.D.

Keille yliopiston portit avautuvat? Tutkimus suomalaisiin yliopistoihin ja eri tieteenaloille valikoitumisesta 2000-luvun alussa

Tutkimuksessa tarkastellaan suomalaisiin yliopistoihin valikoitumista 2000-luvun alussa. Tarkastelu pohjautuu yliopistoon hakeneiden, opiskelemaan hyväksyttyjen ja opiskelupaikkaa ilman jääneiden taustojen vertailuun. Tutkimuksen tarkoituksena on selvittää, miten koulutuksellinen tasa-arvo toteutuu opiskelemaan pääsyssä. Erityistä huomiota kiinnitetään sukupuolten, eri-ikäisten, sosiaalisten ryhmien sekä eri alueella asuvien opiskelijavalinnoissa pärjäämiseen. Lisäksi pohditaan, millaiset taustatekijät ovat yhteydessä opiskelemaan pääsyyn ja miten suomalainen yliopistokenttä on lohkoutunut yliopistoittain ja aloittain hakijoiden ja sisään päässeiden taustojen perusteella. Tutkimuksen pääaineistona on henkilöpohjainen rekisteriaineisto, joka on laadittu valtakunnallisen hakijarekisterin (HAREK) ja Tilastokeskuksen yhteistyönä. Aineisto käsittää 40 %:n satunnaisotoksen vuonna 2003 suomalaisiin yliopistoihin hakeneista (N = 55 790). Aineiston muuttujat kuvaavat hakijoiden taustoja, elämäntilannetta, aiempaa koulutusta ja lapsuudenperheen asemaa. Tutkimuksessa hyödynnetään lisäksi kokonaisjoukosta muodostettua taulukkoaineistoa (N = 139 668). Yliopistoihin hakevat eivät ole yhtenäinen ryhmä. Vaikka suurin osa hakijoista oli nuoria, oli joukossa myös varttuneempia hakijoita, jotka olivat ehtineet hankkia koulutusta ja muuta elämänkokemusta. Päävalinnat toimivat siten myös aikuishakijoiden hakuväylänä; erillisvalintoja eivät hyödynnä läheskään kaikki, joilla siihen olisi mahdollisuus. Klusterianalyysin avulla hakijoista voitiin erottaa neljä ryhmää: 1) nuoret ylioppilaat, 2) toisen tutkinnon suorittajat, 3) koulutuspääoman kartuttajat sekä 4) aikuiset lisäkouluttautujat. Opiskelemaan pääsyyn vaikuttavia tekijöitä analysoitiin logistisen regressioanalyysin avulla. Analyysin mukaan hakijan iällä oli muista taustatekijöistä riippumaton vaikutus opiskelemaan pääsyyn niin, että todennäköisyys päästä yliopistoon vähenee hakijan iän kohotessa. Parhaiten opiskelemaan pääsivät kaikkein nuorimmat, alle 20-vuotiaat hakijat, jotka siis useimmiten ovat saman kevään ylioppilaita. Vanhemmille hakijoille oli usein kertynyt jo koulutusta, mutta aiemmat tutkinnot paransivat sisäänpääsyn mahdollisuuksia vain, mikäli ne olivat korkea-asteelta. Alemmilla ammatillisilla tutkinnoilla oli pikemminkin opiskelemaan pääsyä heikentävä vaikutus. Myös se, mitä hakija oli tehnyt ennen valintakokeita, vaikutti sisäänpääsyn mahdollisuuksiin. Parhaiten valinnoissa pärjäsivät päätoimiset opiskelijat, heikoiten työttömät hakijat. Vaikka miesten hyväksymisprosentit olivat keskimäärin korkeammat kuin naisten, sukupuoli ei osoittautunut itsenäiseksi opiskelemaan pääsyä selittäväksi tekijäksi. Naisten huonompi pärjääminen valinnoissa selittyykin pitkälti sukupuolten eriytyneillä alavalinnoilla. Naisten suosimat alat kun ovat pääsääntöisesti vaikeapääsyisempiä kuin miesten. Tutkimuksessa selvisi myös, että kaupunkilaisuus lisäsi todennäköisyyttä tulla hyväksytyksi. Toisaalta opiskelemaan pääsy erosi myös asuinmaakunnittain, mikä kertoo lähinnä siitä, että eri yliopistojen sisäänpääsyasteissa on varsin suuria eroja. Yliopistojen lohkoutuminen hakijoiden sosiaalisen taustan mukaan oli paljon selvempää kuin alojen. Kaikki pääkaupunkiseudun yliopistot – lukuun ottamatta Teatterikorkeakoulua – luokittuivat isän asemalla mitaten elitistisiksi. Matalimmista taustoista haettiin Lapin, Joensuun ja Vaasan yliopistoihin. Alojen paikka elitistisyyskansanomaisuus -ulottuvuudella vaihteli suuresti yliopistoittain. Teknillistieteellinen, matemaattis-luonnontieteellinen ja kauppatieteellinen ala sijoittuivat kuitenkin keskimääräistä ylemmäs, kun taas kasvatustiede ja farmasia olivat kansanomaisimpia hakukohteita. Opiskelijaksi valikoitumisen peruselementit toistuivat myös tässä tutkimuksessa: koulutetuimpien ja hyvässä asemassa olevien vanhempien jälkeläiset saivat opiskelupaikan useammin kuin muut. Yliopistolaitoksessa vuosikymmenten saatossa toteutetut rakenteelliset muutokset eivät siis ole muuttaneet valikoitumisen peruslinjaa, joskin uutena huomiona nousi maanviljelijöiden jälkeläisten hyvä valinnoissa pärjääminen. Maanviljelijäperheestä tulevien opiskelemaan pääsyn todennäköisyys oli kaikkein suurin.

Ultrassonografia modo B e Doppler na avaliação renal de cães após administração intravenosa de meio de contraste iodado: validação da técnica

Resumo: Meios de contraste iodado podem promover efeitos hemodinâmicos relacionados à vasoconstrição intrarrenal prolongada e redução da perfusão, predispondo à hipóxia e isquemia medular. Alterações de resistência vascular renal podem representar os primeiros sinais de mudança funcional desse órgão. A técnica Doppler pulsado é considerada acessível, não invasiva e permite avaliar a dinâmica vascular dos rins, por meio da aferição dos índices de resistividade (IR) e pulsatilidade (IP). Contudo, na espécie canina, a aquisição de traçados espectrais pode ser penosa devido às dificuldades de varredura e captação de sinal Doppler, sobretudo em relação ao rim direito, devido à sua localização dorsocranial na cavidade abdominal, o que prolonga substancialmente a realização do exame. O objetivo deste estudo é comprovar que a avaliação Doppler pulsado das artérias intrarrenais do rim esquerdo de cães representa a repercussão hemodinâmica renal da administração intravenosa de meios de contraste iodado não sendo necessária a realização do exame nos dois rins. Foram avaliados ambos os rins de seis cadelas adultas em quatro momentos distintos: antes da infusão intravenosa do contraste radiológico e após 1,5 horas, 24 horas e 48 horas, por meio da análise subjetiva da morfologia, ecogenicidade cortical e grau de perfusão renais e análise objetiva do comprimento, volume e resistência vascular intrarrenais (IR e IP). Os parâmetros avaliados ao modo B e Doppler dos rins direito e esquerdo não apresentaram diferenças estatisticamente significativas entre si em cada momento avaliado. Assim, constatou-se que o exame ultrassonográfico Doppler pulsado do rim esquerdo representou a repercussão hemodinâmica renal da aplicação intravenosa de meios de contraste iodado, desde que morfometria, morfologia, ecogenicidade cortical e perfusão de ambos os rins fossem consideradas semelhantes na abordagem ultrassonográfica inicial.

Preparation and purification of Flavobacterium heparinum chondroitinases AC and B by hydrophobic interaction chromatography

Flavobacterium heparinum is a soil bacterium that produces several mucopolysaccharidases such as heparinase, heparitinases I and II, and chondroitinases AC, B, C and ABC. The purpose of the present study was to optimize the preparation of F. heparinum chondroitinases, which are very useful tools for the identification and structural characterization of chondroitin and dermatan sulfates. We observed that during the routine procedure for cell disruption (ultrasound, 100 kHz, 5 min) some of the chondroitinase B activity was lost. Using milder conditions (2 min), most of the chondroitinase B and AC protein was solubilized and the enzyme activities were preserved. Tryptic soy broth without glucose was the best culture medium both for bacterial growth and enzyme induction. Chondroitinases AC and B were separated from each other and also from glucuronidases and sulfatases by hydrophobic interaction chromatography on HP Phenyl-Sepharose. A rapid method for screening of the column fractions was also developed based on the metachromatic shift of the color of dimethylmethylene blue.