Evidence for a modulation of neutral trehalase activity by Ca2+ and cAMP signaling pathways in Saccharomyces cerevisiae

Saccharomyces cerevisiae neutral trehalase (encoded by NTH1) is regulated by cAMP-dependent protein kinase (PKA) and by an endogenous modulator protein. A yeast strain with knockouts of CMK1 and CMK2 genes (cmk1cmk2) and its isogenic control (CMK1CMK2) were used to investigate the role of CaM kinase II in the in vitro activation of neutral trehalase during growth on glucose. In the exponential growth phase, cmk1cmk2 cells exhibited basal trehalase activity and an activation ratio by PKA very similar to that found in CMK1CMK2 cells. At diauxie, even though both cells presented comparable basal trehalase activities, cmk1cmk2 cells showed reduced activation by PKA and lower total trehalase activity when compared to CMK1CMK2 cells. To determine if CaM kinase II regulates NTH1 expression or is involved in post-translational modulation of neutral trehalase activity, NTH1 promoter activity was evaluated using an NTH1-lacZ reporter gene. Similar ß-galactosidase activities were found for CMK1CMK2 and cmk1cmk2 cells, ruling out the role of CaM kinase II in NTH1 expression. Thus, CaM kinase II should act in concert with PKA on the activation of the cryptic form of neutral trehalase. A model for trehalase regulation by CaM kinase II is proposed whereby the target protein for Ca2+/CaM-dependent kinase II phosphorylation is not the neutral trehalase itself. The possible identity of this target protein with the recently identified trehalase-associated protein YLR270Wp is discussed.

Use of molecular epidemiology to monitor the nosocomial dissemination of methicillin-resistant Staphylococcus aureus in a University Hospital from 1991 to 2001

Methicillin-resistant Staphylococcus aureus (MRSA) has been the cause of major outbreaks and epidemics among hospitalized patients, with high mortality and morbidity rates. We studied the genomic diversity of MRSA strains isolated from patients with nosocomial infection in a University Hospital from 1991 to 2001. The study consisted of two periods: period I, from 1991 to 1993 and period II from 1995 to 2001. DNA was typed by pulsed-field gel electrophoresis and the similarity among the MRSA strains was determined by cluster analysis. During period I, 73 strains presented five distinctive DNA profiles: A, B, C, D, and E. Profile A was the most frequent DNA pattern and was identified in 55 (75.3%) strains; three closely related and four possibly related profiles were also identified. During period II, 80 (68.8%) of 117 strains showed the same endemic profile A identified during period I, 18 (13.7%) closely related profiles and 18 (13.7%) possibly related profiles and, only one strain presented an unrelated profile. Cluster analysis showed a 96% coefficient of similarity between profile A from period I and profile A from period II, which were considered to be from the same clone. The molecular monitoring of MRSA strains permitted the determination of the clonal dissemination and the maintenance of a dominant endemic strain during a 10-year period and the presence of closely and possibly related patterns for endemic profile A. However, further studies are necessary to improve the understanding of the dissemination of the endemic profile in this hospital.

Comparison of the Polar S810i monitor and the ECG for the analysis of heart rate variability in the time and frequency domains

The aim of the present study was to compare heart rate variability (HRV) at rest and during exercise using a temporal series obtained with the Polar S810i monitor and a signal from a LYNX® signal conditioner (BIO EMG 1000 model) with a channel configured for the acquisition of ECG signals. Fifteen healthy subjects aged 20.9 ± 1.4 years were analyzed. The subjects remained at rest for 20 min and performed exercise for another 20 min with the workload selected to achieve 60% of submaximal heart rate. RR series were obtained for each individual with a Polar S810i instrument and with an ECG analyzed with a biological signal conditioner. The HRV indices (rMSSD, pNN50, LFnu, HFnu, and LF/HF) were calculated after signal processing and analysis. The unpaired Student t-test and intraclass correlation coefficient were used for data analysis. No statistically significant differences were observed when comparing the values analyzed by means of the two devices for HRV at rest and during exercise. The intraclass correlation coefficient demonstrated satisfactory correlation between the values obtained by the devices at rest (pNN50 = 0.994; rMSSD = 0.995; LFnu = 0.978; HFnu = 0.978; LF/HF = 0.982) and during exercise (pNN50 = 0.869; rMSSD = 0.929; LFnu = 0.973; HFnu = 0.973; LF/HF = 0.942). The calculation of HRV values by means of temporal series obtained from the Polar S810i instrument appears to be as reliable as those obtained by processing the ECG signal captured with a signal conditioner.

Use of anti-PGL-1 antibodies to monitor therapy regimes in leprosy patients

The suitability of IgM antibodies to PGL-1 for monitoring the response to multidrug therapy (MDT) was sequentially tested by ELISA in 105 leprosy patients, and bacterial indexes (BI) were also determined. Patients were divided into 3 groups: group 1, 34 multibacillary (MB) patients treated for 12 months with MDT-MB; group 2, 33 MB patients treated for 24 months with MDT-MB, and group 3, 38 paucibacillary (PB) patients treated for 6 months with MDT-PB. Untreated MB patients exhibited higher antibody levels (mean ± SEM): group 1 (6.95 ± 1.35) and group 2 (12.53 ± 2.02) than untreated PB patients (1.28 ± 0.35). There was a significant difference (P < 0.01) in anti-PGL-1 levels in group 1 patients: untreated (6.95 ± 1.35) and treated for 12 months (2.78 ± 0.69) and in group 2 patients: untreated (12.53 ± 2.02) and treated for 24 months (2.62 ± 0.79). There was no significant difference between untreated (1.28 ± 0.35) and treated (0.62 ± 0.12) PB patients. Antibody levels correlated with BI. The correlation coefficient (Pearson’s r) was 0.72 before and 0.23 (P < 0.05) after treatment in group 1 and 0.67 before and 0.96 (P < 0.05) after treatment in group 2. BI was significantly reduced (P < 0.01) after 12 and 24 months on MDT (group 1: 1.26-0.26; group 2: 1.66-0.36). Our data indicate that monitoring anti-PGL-1 levels during MDT may be a sensitive tool for evaluating treatment efficacy. These data also indicate that the control of leprosy infection can be obtained with 12 months of MDT in MB patients.

Retinoic acid and cAMP inhibit rat hepatocellular carcinoma cell proliferation and enhance cell differentiation

Hepatocellular carcinoma (HCC) is the third highest cause of cancer death worldwide. In general, the disease is diagnosed at an advanced stage when potentially curative therapies are no longer feasible. For this reason, it is very important to develop new therapeutic approaches. Retinoic acid (RA) is a natural derivative of vitamin A that regulates important biological processes including cell proliferation and differentiation. In vitro studies have shown that RA is effective in inhibiting growth of HCC cells; however, responsiveness to treatment varies among different HCC cell lines. The objective of the present study was to determine if the combined use of RA (0.1 µM) and cAMP (1 mM), an important second messenger, improves the responsiveness of HCC cells to RA treatment. We evaluated the proliferative behavior of an HCC cell line (HTC) and the expression profile of genes related to cancer signaling pathway (ERK and GSK-3β) and liver differentiation (E-cadherin, connexin 26 (Cx26), and Cx32). RA and cAMP were effective in inhibiting the proliferation of HTC cells independently of combined use. However, when a mixture of RA and cAMP was used, the signals concerning the degree of cell differentiation were increased. As demonstrated by Western blot, the treatment increased E-cadherin, Cx26, Cx32 and Ser9-GSK-3β (inactive form) expression while the expression of Cx43, Tyr216-GSK-3β (active form) and phosphorylated ERK decreased. Furthermore, telomerase activity was inhibited along treatment. Taken together, the results showed that the combined use of RA and cAMP is more effective in inducing differentiation of HTC cells.

Man and the Milieu: Ecocritical analysis of Ernest Hemingway’s “Big Two-Hearted River” and “Indian Camp

Pro Gradu -tutkielmassani tarkastelen Ernest Hemingwayn alun perin vuonna 1925 julkaistuja kahta novellia ”Big Two-Hearted River” ja ”Indian Camp”. Molemmat teokset kuuluvat Nick Adams -tarinoihin, jotka ovat kokoelma novelleja, joissa Hemingway kuvaa Nick Adamsin elämää pojasta mieheksi. Nick Adams on usein tulkittu Hemingwayn alter egoksi ja monet tapahtumat ovatkin suoraa heijastusta Hemingwayn elämästä. Päähenkilön lisäksi monia Nick Adams -tarinoita yhdistää luonnon vahva läsnäolo ja Hemingwayn kuuluisan maalauksellinen miljöön deskriptiivisyys. Käyttämäni ekokriittisen teorian avulla pyrin osoittamaan tutkielmassani, että Nick Adamsin hahmo peilautuu miljöökuvauksessa syventyen dynaamiseksi hahmoksi ja että Hemingwayn luonto- ja ympäristökuvaus peilaa hahmon sisäistä maailmaa hyvinkin läheisesti. Tutkimukseni teoreettinen viitekehys, ekokritiikki, on kirjallisuudentutkimuksen suuntaus, joka tutkii ihmisen ja fyysisen ympäristön suhdetta toisiinsa. Lisäksi hyödynnän tutkielmassani dikotomioita, joiden avulla tarkastelen ja analysoin novelleista nousevia aiheita. Esimerkiksi seuraavat dikotomiat ovat keskeisiä molemmissa novelleissa: kulttuuri vs. luonto, pastoraali vs. erämaa, matka vs. oleskelu, keho vs. mieli, vesi vs. maa, tuli tai valo vs. vesi ja elämä vs. kuolema.

Avantgarde, karnevalismi ja camp Eric Wareheimin musiikkivideoissa

Tutkielman tavoitteena on analysoida ohjaaja Eric Wareheimin musiikkivideoiden sisältämiä visuaalisia representaatioita ja selvittää, onko niillä ilmaisuna vastakulttuurista potentiaalia. Musiikkivideoiden jakelun ja katsomiskokemuksen siirtymä television kaapelikanavilta internetiin videopalvelimien sivuille viimeisen vuosikymmenen aikana, on muuttanut myös musiikkivideoiden kuvallista sisältöä. Kaapelikanavien ohjelmistopolitiikkaan kuuluva sensuuri ei ole enää ilmaisun esteenä internetissä löydettävien indie-videoiden kohdalla, joihin Wareheimin musiikkivideotuotanto pitkälti paikantuu. Wareheimin videoiden vastakulttuurisen merkityksen avaamiseen käytän tutkimuksessani näkemyksiä avantgardesta, karnevalismista ja camp-estetiikasta. Pyrin näillä poliittiseksi katsomillani näkökulmilla selvittämään lähiluvun kautta Wareheimin videoiden kuvallista voimaa vastakulttuurin muotona suhteuttaen niiden sisältöä aikaisempien vuosikymmenten ja nykyhetken valtavirtamusiikkivideoon. Analyysissani paljastuu, että Wareheimin musiikkivideoissa on uusia kuvallisia sisältöjä aikaisempaan valtavirtamusiikkivideon estetiikkaan nähden. Etnisyys, ruumis ja seksuaalisuus representoidaan videoissa kumouksellisella tavalla shokkihuumoriksi kutsuttavan tyylin kautta. Wareheimin musiikkivideot ottavat kantaa myös sosiaaliseen ja yhteiskunnalliseen järjestykseen rikkoen vaatimuksen valtakulttuurin sanelemasta kulttuurituotteiden sisällöllisestä poliittisesta korrektiudesta. Wareheimin musiikkivideot paljastuvat tutkimuksessani yhdenlaisen, populaarikulttuuriin lukeutuvan, vastakulttuurin visuaaliseksi ilmaisuksi.

18/6/20, Satory [Grand prix des chars d'assauts ou Grand steeple des tanks au camp militaire de Satory, Versailles] : [photographie de presse] / [Agence Rol]

Référence bibliographique : Rol, 59741

Camp britannique à Constantinople : [photographie de presse] / [Agence Rol]

Référence bibliographique : Rol, 59801

[Lettre de M. le Chevalier de Soupire envoyée du camp de Valence le 2 septembre 1755]

Ancien possesseur : Argenson, Antoine-René de Voyer (1722-1787 ; marquis de Paulmy d')

La promenade des bourgeois de Paris au camp de Ville-Juifve, en vers burlesques

[Mazarinade. 1649]

Journal du Camp de Gray commandé par M. Le duc de Randan

Ancien possesseur : Argenson, Antoine-René de Voyer (1722-1787 ; marquis de Paulmy d')

[Journal du Camp de Gray commandé par M. Le duc de Randan]. Ordre du 26. au 27. aoust

Ancien possesseur : Argenson, Antoine-René de Voyer (1722-1787 ; marquis de Paulmy d')