Impairment of cytomegalovirus-specific cellular immune response as a risk factor for cytomegalovirus disease in transplant recipients

Human cytomegalovirus (CMV) infection is common in most people but nearly asymptomatic in immunocompetent individuals. After primary infection the virus persists throughout life in a latent form in a variety of tissues, particularly in precursor cells of the monocytic lineage. CMV reinfection and occurrence of disease are associated with immunosuppressive conditions. Solid organ and bone marrow transplant patients are at high risk for CMV disease as they undergo immunosuppression. Antiviral treatment is effective in controlling viremia, but 10-15% of infected patients can experience CMV disease by the time the drug is withdrawn. In addition, long-term antiviral treatment leads to bone marrow ablation and renal toxicity. Furthermore, control of chronic CMV infection in transplant recipients appears to be dependent on the proper recovery of cellular immunity. Recent advances in the characterization of T-cell functions and identification of distinct functional signatures of T-cell viral responses have opened new perspectives for monitoring transplant individuals at risk of developing CMV disease.

Imbalance of naive and memory T lymphocytes with sustained high cellular activation during the first year of life from uninfected children born to HIV-1-infected mothers on HAART

The immune consequences of in utero HIV exposure to uninfected children whose mothers were submitted to highly active antiretroviral therapy (HAART) during gestation are not well defined. We evaluated 45 HIV-exposed uninfected (ENI) neonates and 45 healthy unexposed control (CT) neonates. All HIV-infected mothers received HAART during pregnancy, and the viral load at delivery was <50 copies/mL for 56.8%. Twenty-three ENI neonates were further evaluated after 12 months and compared to 23 unexposed healthy age-matched infants. Immunophenotyping was performed by flow cytometry in cord and peripheral blood. Cord blood lymphocyte numbers did not differ between groups. However, ENI neonates had a lower percentage of naive T cells than CT neonates (CD4+, 76.6 vs 83.1%, P < 0.001; CD8+, 70.9 vs 79.6%, P = 0.003) and higher percentages of central memory T cells than CT neonates (CD4+, 13.9 vs 8.7%, P < 0.001; CD8+, 8.6 vs 4.8%, P = 0.001). CD38 mean fluorescence intensity of T cells was higher in ENI neonates (CD4+, 62.2 vs 52.1, P = 0.007; CD8+, 47.7 vs 35.3, P < 0.001). At 12 months, ENI infants still had higher mean fluorescence intensity of CD38 on T cells (CD4+, 34.2 vs 23.3, P < 0.001; CD8+, 26.8 vs 19.4, P = 0.035). Despite effective maternal virologic control at delivery, HIV-exposed uninfected children were born with lower levels of naive T cells. Immune activation was present at birth and remained until at least 12 months of age, suggesting that in utero exposure to HIV causes subtle immune abnormalities.

Humoral and cellular immune responses to Blomia tropicalis and concanavalin A-binding fractions in atopic patients

Blomia tropicalis, Dermatophagoides pteronyssinus and D. farinae are prevalent house dust mites. Concanavalin A-binding components derived from B. tropicalis (Bt-ConA extract) are highly immunogenic in allergic diseases. The aim of the present study was to evaluate the humoral and cellular immune responses to B. tropicalis in mite-sensitized patients. A total of 137 patients with allergic rhinitis with/without asthma and 109 non-atopic subjects were selected and analyzed by the skin prick test, and for total serum IgE and specific IgE levels to both Bt-total and Bt-ConA extracts, their proliferative response and cytokine (IFN-γ and IL-5) production by peripheral blood mononuclear cells (PBMC) stimulated with both extracts. Skin prick test showed that 70% of the patients were sensitized to Bt (Bt+) and similar levels of specific IgE to Bt-total and Bt-ConA extracts were demonstrable in Bt+ patients. Significant PBMC proliferation was observed in response to Bt-total extract in Bt+, but not in Bt- patients and non-atopic subjects (P < 0.001). Bt-ConA extract induced increased proliferative responses in all patient groups compared to medium alone (P < 0.05), but these responses were significantly decreased in the presence of the mannopyranoside ConA inhibitor (P < 0.05). Significant IFN-γ production was observed after Bt-ConA stimulation of Bt+ patients (P < 0.05), while Bt-total extract had no effect. IL-5 production was consistently detected in Bt+ patients after allergen-specific stimulation or with no stimulus, indicating that PBMC from allergic patients are prone to produce Th2 profile cytokines, spontaneously or inductively by allergen restimulation. These data showed that ConA-binding components isolated from B. tropicalis may contain relevant antigens that are involved in both humoral and cellular immune responses. However, without an additional purification procedure to eliminate the residual contamination with ConA, its use in immunotherapeutic procedures cannot be recommended.

Molecular and cellular pathogenesis of autosomal dominant polycystic kidney disease

Autosomal dominant polycystic kidney disease (ADPKD) is one of the most common human life-threatening monogenic disorders. The disease is characterized by bilateral, progressive renal cystogenesis and cyst and kidney enlargement, often leading to end-stage renal disease, and may include extrarenal manifestations. ADPKD is caused by mutation in one of two genes, PKD1 and PKD2, which encode polycystin-1 (PC1) and polycystin-2 (PC2), respectively. PC2 is a non-selective cation channel permeable to Ca2+, while PC1 is thought to function as a membrane receptor. The cyst cell phenotype includes increased proliferation and apoptosis, dedifferentiation, defective planar polarity, and a secretory pattern associated with extracellular matrix remodeling. The two-hit model for cyst formation has been recently extended by the demonstration that early gene inactivation leads to rapid and diffuse development of renal cysts, while inactivation in adult life is followed by focal and late cyst formation. Renal ischemia/reperfusion, however, can function as a third hit, triggering rapid cyst development in kidneys with Pkd1 inactivation induced in adult life. The PC1-PC2 complex behaves as a sensor in the primary cilium, mediating signal transduction via Ca2+ signaling. The intracellular Ca2+ homeostasis is impaired in ADPKD, being apparently responsible for the cAMP accumulation and abnormal cell proliferative response to cAMP. Activated mammalian target for rapamycin (mTOR) and cell cycle dysregulation are also significant features of PKD. Based on the identification of pathways altered in PKD, a large number of preclinical studies have been performed and are underway, providing a basis for clinical trials in ADPKD and helping the design of future trials.

De radio aeternitatis seu honestate e suo fonte scilicet jure naturae, emanante

Variantti B.

Cellular and molecular studies of the effects of a selective COX-2 inhibitor celecoxib in the cardiac cell line H9c2 and their correlation with death mechanisms

Cardiovascular disease is one of the leading causes of death worldwide, and evidence indicates a correlation between the inflammatory process and cardiac dysfunction. Selective inhibitors of cyclooxygenase-2 (COX-2) enzyme are not recommended for long-term use because of potentially severe side effects to the heart. Considering this and the frequent prescribing of commercial celecoxib, the present study analyzed cellular and molecular effects of 1 and 10 µM celecoxib in a cell culture model. After a 24-h incubation, celecoxib reduced cell viability in a dose-dependent manner as also demonstrated in MTT assays. Furthermore, reverse transcription-polymerase chain reaction analysis showed that the drug modulated the expression level of genes related to death pathways, and Western blot analyses demonstrated a modulatory effect of the drug on COX-2 protein levels in cardiac cells. In addition, the results demonstrated a downregulation of prostaglandin E2 production by the cardiac cells incubated with celecoxib, in a dose-specific manner. These results are consistent with the decrease in cell viability and the presence of necrotic processes shown by Fourier transform infrared analysis, suggesting a direct correlation of prostanoids in cellular homeostasis and survival.

Oxidative stress and cellular immunity in patients with recurrent aphthous ulcers

Recurrent aphthous ulcer (RAU) is an inflammatory condition of the oral mucosa characterized by painful, well-circumscribed, single or multiple round or ovoid ulcerations. The exact etiologic factor(s) of these ulcerations are not yet understood. The objective of this study was to evaluate inflammatory processes and free radical metabolism of 25 patients with RAUs compared to 25 healthy controls. The levels of malondialdehyde (MDA) and glutathione (GSH) were determined by high-performance liquid chromatography. Tumor necrosis factor-alpha (TNF-α), interleukin-2 (IL-2), IL-10, and IL-12 were determined by ELISA. Nitric oxide (NO), myeloperoxidase (MPO), total antioxidant status (TAS), and total oxidant status (TOS) levels were measured spectroscopically in serum. The levels of MDA, GSH, TNF-α, IL-2, IL-12, MPO, and TOS, and oxidative stress index (OSI) were higher, and the levels of NO, IL-10, and TAS were lower in patients with RAU than in controls. Statistical analysis showed that GSH, TNF-α, IL-2, IL-10, and OSI differed significantly in patients with RAU compared to controls. These parameters have important roles in oxidant/antioxidant defense.

Therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia using Fe2O3 nanoparticles

This study aimed to investigate the therapeutic mechanism of treating SMMC-7721 liver cancer cells with magnetic fluid hyperthermia (MFH) using Fe2O3 nanoparticles. Hepatocarcinoma SMMC-7721 cells cultured in vitro were treated with ferrofluid containing Fe2O3 nanoparticles and irradiated with an alternating radio frequency magnetic field. The influence of the treatment on the cells was examined by inverted microscopy, MTT and flow cytometry. To study the therapeutic mechanism of the Fe2O3 MFH, Hsp70, Bax, Bcl-2 and p53 were detected by immunocytochemistry and reverse transcription polymerase chain reaction (RT-PCR). It was shown that Fe2O3 MFH could cause cellular necrosis, induce cellular apoptosis, and significantly inhibit cellular growth, all of which appeared to be dependent on the concentration of the Fe2O3 nanoparticles. Immunocytochemistry results showed that MFH could induce high expression of Hsp70 and Bax, decrease the expression of mutant p53, and had little effect on Bcl-2. RT-PCR indicated that Hsp70 expression was high in the early stage of MFH (<24 h) and became low or absent after 24 h of MFH treatment. It can be concluded that Fe2O3 MFH significantly inhibited the proliferation of in vitro cultured liver cancer cells (SMMC-7721), induced cell apoptosis and arrested the cell cycle at the G2/M phase. Fe2O3 MFH can induce high Hsp70 expression at an early stage, enhance the expression of Bax, and decrease the expression of mutant p53, which promotes the apoptosis of tumor cells.

De radio aeternitatis seu honestate e suo fonte scilicet jure naturae, emanante

Variantti A.

The effects of mutated cationic amino acid transporter y+LAT1 at the cellular and systemic level

y+LAT1 is a transmembrane protein that, together with the 4F2hc cell surface antigen, forms a transporter for cationic amino acids in the basolateral plasma membrane of epithelial cells. It is mainly expressed in the kidney and small intestine, and to a lesser extent in other tissues, such as the placenta and immunoactive cells. Mutations in y+LAT1 lead to a defect of the y+LAT1/4F2hc transporter, which impairs intestinal absorbance and renal reabsorbance of lysine, arginine and ornithine, causing lysinuric protein intolerance (LPI), a rare, recessively inherited aminoaciduria with severe multi-organ complications. This thesis examines the consequences of the LPI-causing mutations on two levels, the transporter structure and the Finnish patients’ gene expression profiles. Using fluorescence resonance energy transfer (FRET) confocal microscopy, optimised for this work, the subunit dimerisation was discovered to be a primary phenomenon occurring regardless of mutations in y+LAT1. In flow cytometric and confocal microscopic FRET analyses, the y+LAT1 molecules exhibit a strong tendency for homodimerisation both in the presence and absence of 4F2hc, suggesting a heterotetramer for the transporter’s functional form. Gene expression analysis of the Finnish patients, clinically variable but homogenic for the LPI-causing mutation in SLC7A7, revealed 926 differentially-expressed genes and a disturbance of the amino acid homeostasis affecting several transporters. However, despite the expression changes in individual patients, no overall compensatory effect of y+LAT2, the sister y+L transporter, was detected. The functional annotations of the altered genes included biological processes such as inflammatory response, immune system processes and apoptosis, indicating a strong immunological involvement for LPI.

Kognitiivinen radio sotilaallisen maanpuolustuksen kontekstissa

Kognitiiviteknologialla on ennustettu olevan disruptiivisia vaikutuksia sodankäyntiin. Kognitiiviteknologian avulla pyritään löytämään myös ratkaisu kasvavista datansiirtovaatimuksista aiheutuvaan radiotaajuisen spektrin ahtauteen. Tämän tutkimuksen keskiössä on kognitiivinen radio. Diplomityössä ennakoidaan niitä muutoksia, joita radion kognitiiviset ominaisuudet tuovat sotilaalliseen toimintaympäristöön. Tutkimuksessa etsitään kognitiivisen radion suorituskykyyn, suorituskyvyn rakentamiseen ja käyttöön liittyviä tekijöitä, jotka ovat merkityksellisiä sotilaallisesta näkökulmasta. Radion kognitiivisuutta käsitellään kongitiivisten ominaisuuksien, kuten adaptiivisuuden tai tilannetietoisuuden, kautta. Painopisteenä on kognitiivisten ominaisuuksien keskinäinen arvottaminen, mahdollisten vaikutusten tarkastelu operatiivisessa viitekehyksessä sekä tarvittavien toimenpiteiden kartoittaminen kognitiivisen tekniikan operatiivisen käytön mahdollistamiseksi. Tutkimuksen pohjalta esitetään kognitiivisten ominaisuuksien tärkeysjärjestys suorituskyvyn ja johtamisjärjestelmien kehittämisen näkökulmista ja alustavia operatiivisia vaatimuksia kognitiiviselle radiolle sekä kuvataan spektrin hallinnan kehittämismahdollisuuksia. Operatiivisia vaikutuksia tutkitaan myös uhkalähtöisesti eli arvioimalla, miten tietty kognitiivinen ominaisuus vaikuttaisi yleisimpiin tiedonsiirtoverkkoihin kohdistuviin uhkiin. Lisäksi työssä arvioidaan kognitiivisen radion käyttöönoton esteitä Puolustusvoimissa. Tutkimuksen päämenetelmä on delfoi, jota on täydennetty asiantuntijahaastatteluin sekä -arvioin. Kognitiiviselle radiolle on useita erilaisia määritelmiä. Kaikkia kriteerejä täyttävää kognitiivista radiota ei ole vielä olemassa, vaikka kognitiivisia ominaisuuksia radioissa on ollut pitkään. Kognitiivinen radio, tai jo pelkästään ohjelmistoradio kehittyneillä kognitiivisilla ominaisuuksilla, tuo huomattavia mahdollisuuksia parantaa langattoman tiedonsiirron toimintakykyä sotilaallisessa käyttöympäristössä. Merkittävin suorituskykylisä sotilaallisessa käyttöympäristössä saadaan seuraavista kognitiivisista ominaisuuksista: 1) dynaaminen spektrin hyväksikäyttö (DSA) 2) yhteyksien adaptiivisuus ja radioresurssien hallinta (SLA ja RRM) sekä 3) älykäs verkonmuodostus (SON ja RBR). Tiedonsiirtojärjestelmien ylläpito ja rakentaminen vaativat aiempaa enemmän suunnittelua ja ammattitaitoa, mutta toisaalta loppukäyttäjän osaamistasovaatimuksia voidaan laskea sitä mukaa, kun lähestytään 0-konfiguraatiojärjestelmiä. Radioiden kognitiivisuuden myötä voidaan myös passiivisiin uhkiin, esimerkiksi elektroniseen tiedusteluun, varautua aiempaa tehokkaammin. Aktiivisiin uhkiin, kuten häirintään, kognitiiviset ominaisuudet vaikuttavat jopa toiminnan fundamentteja muuttavasti. Tiedonsiirtojärjestelmien ohjelmistopohjaisuus nostaa kyberin merkitttävyyttä arvioitaessa eri uhkien kokonaismerkityksiä. Onnistunut kyberoperaatio voi tarkoittaa koko järjestelmän lamautumista tai vakavaa tietovuotoa. Kognitiivisten ominaisuuksien liittäminen osaksi sotilasradiojärjestelmiä on haasteellista. Käyttöönoton esteitä ovat osaamisen puute, lainsäädännön ja regulaation jäykkyys, tekniikan osittainen kypsymättömyys sekä tietoturvan hallinta. Spektri on muutoksessa, mikä tulee näkymään spektrin sirpaloitumisena vuoteen 2020 mennessä. Paine taajuushallinnan muuttamisesta kognitiivitekniikkaa sallivampaan suuntaan nousee erityisesti siviilipuolen kehittyvien mobiililaitteiden datansiirtomäärien kasvaessa. Lainsäädäntö antaa viranomai-sille suuret valtuudet säädellä spektrin käyttöä poikkeusoloissa. Tämä mahdollistaa sotilaallisten järjestelmien suunnittelun ja kehittämisen taajuusriippumattomasti jo normaalioloissa. Haasteena on spektrin hallinnoinnin kehittäminen esimerkiksi ennakoitua nopeammassa tilannekehityksessä. Spektriä tulisi hallita kokonaisturvallisuuden näkökulmasta joustavasti viranomaisten yhteisillä päätöksillä. Kognitiivisen radion tulisi tulevaisuudessa voida käyttää muille järjestelmille osoitettuja taajuusresursseja taajuuksien ollessa käyttämättömänä. Kognitiivista radiota kannattaisi kehittää taajuusvapaasti. Spektri tullee olemaan tulevaisuudessa yhä ahtaampi, ja tähän haasteeseen kognitiivinen radio tarjoaa osaltaan ratkaisun.

Citizen Jane : exploring the relationship between gender and cellular phones in societies of control

In this thesis, I argue that the mutually productive relationship between women (as gendered subjects) and cellular phone technology is one of control. Women use cellular phones to organize, manage and otherwise control the multiplicity of tasks required of them on a daily basis. At the same time, through using cell phones, women participate in regimes of control including surveillance and persistent connection. I explore this relationship at the level of everyday practice, and conclude by speculating about this relationship at a wider level of social control and organization. This argument emerges from the critical approach suggested by Slack and Wise (2005), who argue that technology and culture are inseparable. They provide articulations and assemblages as tools of analysis. I situate this analysis more broadly within Foucault's (1991) work on govemmentality, in its modem form of societies of control (Deleuze, 1995b).

Response curves of deterministic and probabilistic cellular automata in one and two dimensions

One of the most important problems in the theory of cellular automata (CA) is determining the proportion of cells in a specific state after a given number of time iterations. We approach this problem using patterns in preimage sets - that is, the set of blocks which iterate to the desired output. This allows us to construct a response curve - a relationship between the proportion of cells in state 1 after niterations as a function of the initial proportion. We derive response curve formulae for many two-dimensional deterministic CA rules with L-neighbourhood. For all remaining rules, we find experimental response curves. We also use preimage sets to classify surjective rules. In the last part of the thesis, we consider a special class of one-dimensional probabilistic CA rules. We find response surface formula for these rules and experimental response surfaces for all remaining rules.