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  • Gruzdaitis, Päivi (2011)
    Complications of atherosclerosis such as myocardial infarction and stroke are the primary cause of death in Western societies. The development of atherosclerotic lesions is a complex process, including endothelial cell dysfunction, inflammation, extracellular matrix alteration and vascular smooth muscle cell (VSMC) proliferation and migration. Various cell cycle regulatory proteins control VSMC proliferation. Protein kinases called cyclin dependent kinases (CDKs) play a major role in regulation of cell cycle progression. At specific phases of the cell cycle, CDKs pair with cyclins to become catalytically active and phosphorylate numerous substrates contributing to cell cycle progression. CDKs are also regulated by cyclin dependent kinase inhibitors, activating and inhibitory phosphorylation, proteolysis and transcription factors. This tight regulation of cell cycle is essential; thus its deregulation is connected to the development of cancer and other proliferative disorders such as atherosclerosis and restenosis as well as neurodegenerative diseases. Proteins of the cell cycle provide potential and attractive targets for drug development. Consequently, various low molecular weight CDK inhibitors have been identified and are in clinical development. Tylophorine is a phenanthroindolizidine alkaloid, which has been shown to inhibit the growth of several human cancer cell lines. It was used in Ayurvedic medicine to treat inflammatory disorders. The aim of this study was to investigate the effect of tylophorine on human umbilical vein smooth muscle cell (HUVSMC) proliferation, cell cycle progression and the expression of various cell cycle regulatory proteins in order to confirm the findings made with tylophorine in rat cells. We used several methods to determine our hypothesis, including cell proliferation assay, western blot and flow cytometric cell cycle distribution analysis. We demonstrated by cell proliferation assay that tylophorine inhibits HUVSMC proliferation dose-dependently with an IC50 value of 164 nM ± 50. Western blot analysis was used to determine the effect of tylophorine on expression of cell cycle regulatory proteins. Tylophorine downregulates cyclin D1 and p21 expression levels. The results of tylophorine’s effect on phosphorylation sites of p53 were not consistent. More sensitive methods are required in order to completely determine this effect. We used flow cytometric cell cycle analysis to investigate whether tylophorine interferes with cell cycle progression and arrests cells in a specific cell cycle phase. Tylophorine was shown to induce the accumulation of asynchronized HUVSMCs in S phase. Tylophorine has a significant effect on cell cycle, but its role as cell cycle regulator in treatment of vascular proliferative diseases and cancer requires more experiments in vitro and in vivo.
  • Aaltonen, Kirsimari (Helsingin yliopisto, 2008)
    Breast cancer is the most common malignancy in women in Western countries. It is a heterogeneous disease with varying biological characteristics and aggressiveness. Family history is one of the strongest predisposing factors for breast cancer. The known susceptibility genes explain only around 25% of all familial breast cancers. At least part of the unknown familial aggregation may be caused by several low-penetrance variants that occur commonly in the general population. Cyclins are cell cycle-regulating proteins. Cyclin expression oscillates during the cell cycle and is under strict control. In cancer cells, cyclin expression often becomes deregulated, leading to uncontrolled cell division and proliferation, one of the hallmarks of cancer. In this study, we investigated the role of cyclins in breast cancer predisposition, pathogenesis, and tumor behavior. Cyclin A immunohistochemistry was evaluated both on traditional large sections and on tissue microarrays (TMA). The concordance of the results was good, indicating that TMA is a reliable method for studying cyclin expression in breast cancer. The expression of cyclins D1, E, and B1 was studied among 1348 invasive breast cancers on TMA. Familial BRCA1/2-mutation negative tumors had significantly more often low cyclin E and high cyclin D1 expression than BRCA1/2 related or sporadic tumors. Unique cyclin E and D1 expression patterns among familial non-BRCA1/2 breast cancers may reflect different predisposition and pathogenesis in these groups and help to differentiate mutation-positive from mutation-negative familial cancers. High cyclin E expression was associated with an aggressive breast cancer phenotype and was an independent marker of poor metastasis-free survival. High cyclin D1 was associated with high grade and high proliferation among estrogen receptor (ER)-positive but with low grade and low proliferation among ER-negative breast cancers. Among ER-positive cancers not treated with chemotherapy, high cyclin D1 showed a trend towards shorter metastasis-free survival. These results suggest that different mechanisms may drive proliferation in ER-negative and -positive breast cancers and that cyclin D1 has a particularly important role in tumorigenesis of hormone receptor-positive breast cancer. High cyclin B1 expression was associated with aggressive breast cancer features and had an independent impact on survival. The results suggest that cyclin B1 immunohistochemistry is a method that could easily be adapted for routine use and is an independent prognostic factor, adding specificity to prognostic evaluation conducted with traditional markers. A commonly occurring cyclin D1 gene polymorphism A870G was associated with increased breast cancer risk in a large material of Finnish and Canadian breast cancer patients. The interaction of the high-activity alleles of cyclin D1 gene and estrogen metabolism gene COMT conferred an even higher risk. These results show that cyclin D1 and COMT act synergistically to contribute to breast cancer progression and that individual risk for breast cancer can be altered by the combined effect of polymorphisms with low-penetrance alleles. By investigating critical cell cycle regulator protein cyclins, we revealed new aspects of breast cancer predisposition, pathogenesis, and clinical course.
  • Erkinheimo, Tiina-Liisa (Helsingin yliopisto, 2005)
  • Saukkonen, Kirsi (Helsingin yliopisto, 2003)
  • Sairanen, Jukka (Jukka Sairanen, 2008)
    Painful bladder syndrome/interstitial cystitis (PBS/IC) is a chronic urinary bladder disorder of unknown etiology characterized by symptoms of bladder pain and urinary frequency. PBS/IC is a chronic disease in which drug therapy has not led to significant success over the course of time. If the symptoms of PBS/IC are refractory to standard treatments, a possible cure might demand surgical intervention involving cystectomy. The eventual autoimmune etiology in mind, immunosuppressive drug therapy with cyclosporine A (CyA) was started to patients with refractory PBS/IC. CyA is a potent anti-inflammatory drug, a calcineurin inhibitor which inhibits T lymphocyte IL-2 produc-tion. T cells are present in abundance in inflammation of the bladder in PBS/IC. On the basis of a pilot, short-term study with CyA on PBS/IC, use of CyA was continued empirically over the long term. We conducted a prospective, randomized, six-month study in 64 patients comparing the effect of CyA with the FDA approved treatment, pentosan polysulfate sodium (PPS). We measured the drug effect on patient s symptoms, the potassium sensitivity test, and on urinary biomarkers. We further tested the impact of CyA, PPS, DMSO and BCG therapy on a health-related quality of life questionnaire and evaluated the response rate to treatment with these therapies. Long-term use of CyA was safe and effective in PBS/IC patients. The good clinical effect matured individually during the years in which CyA was continued. Cessation of medication led to the reappearance of symptoms, and restarting CyA to renewed alleviation, so that CyA was administered as continuous medication. The response rate to CyA increased during the study period, comprising 75% of CyA patients at six months. 19% of patients responded to PPS therapy. Adverse effects were more common in the CyA group, underlining the importance of monitoring the drug safety and appropriate titration of the dose. The potassium sensitivity test is positive in the majority of PBS/IC patients. Successful therapy of PBS/IC can alter nerve sensitivity to external potassium. This effect was seen more often after CyA therapy. Successful treatment of PBS/IC with CyA resulted to decreasing urinary levels of EGF. IL-6 levels in urine were higher among older patient with a longer history of PBS/IC. In these patients, reduced levels of urinary IL-6 were measured after CyA therapy. Patients who experience the best treatment response have improved quality of life according to the post-treatment health-related quality of life (HRQOL) questionnaire. CyA had more impact on the ma-jority of the aspects of QoL than PPS. Despite DMSO therapy being more successful than BCG in the count of responders, DMSO and BCG had equal effects on the HRQOL questionnaire.
  • Fanta, Samuel (Helsingin yliopisto, 2009)
    Cyclosporine is an immunosuppressant drug with a narrow therapeutic index and large variability in pharmacokinetics. To improve cyclosporine dose individualization in children, we used population pharmacokinetic modeling to study the effects of developmental, clinical, and genetic factors on cyclosporine pharmacokinetics in altogether 176 subjects (age range: 0.36–20.2 years) before and up to 16 years after renal transplantation. Pre-transplantation test doses of cyclosporine were given intravenously (3 mg/kg) and orally (10 mg/kg), on separate occasions, followed by blood sampling for 24 hours (n=175). After transplantation, in a total of 137 patients, cyclosporine concentration was quantified at trough, two hours post-dose, or with dose-interval curves. One-hundred-four of the studied patients were genotyped for 17 putatively functionally significant sequence variations in the ABCB1, SLCO1B1, ABCC2, CYP3A4, CYP3A5, and NR1I2 genes. Pharmacokinetic modeling was performed with the nonlinear mixed effects modeling computer program, NONMEM. A 3-compartment population pharmacokinetic model with first order absorption without lag-time was used to describe the data. The most important covariate affecting systemic clearance and distribution volume was allometrically scaled body weight i.e. body weight**3/4 for clearance and absolute body weight for volume of distribution. The clearance adjusted by absolute body weight declined with age and pre-pubertal children (< 8 years) had an approximately 25% higher clearance/body weight (L/h/kg) than did older children. Adjustment of clearance for allometric body weight removed its relationship to age after the first year of life. This finding is consistent with a gradual reduction in relative liver size towards adult values, and a relatively constant CYP3A content in the liver from about 6–12 months of age to adulthood. The other significant covariates affecting cyclosporine clearance and volume of distribution were hematocrit, plasma cholesterol, and serum creatinine, explaining up to 20%–30% of inter-individual differences before transplantation. After transplantation, their predictive role was smaller, as the variations in hematocrit, plasma cholesterol, and serum creatinine were also smaller. Before transplantation, no clinical or demographic covariates were found to affect oral bioavailability, and no systematic age-related changes in oral bioavailability were observed. After transplantation, older children receiving cyclosporine twice daily as the gelatine capsule microemulsion formulation had an about 1.25–1.3 times higher bioavailability than did the younger children receiving the liquid microemulsion formulation thrice daily. Moreover, cyclosporine oral bioavailability increased over 1.5-fold in the first month after transplantation, returning thereafter gradually to its initial value in 1–1.5 years. The largest cyclosporine doses were administered in the first 3–6 months after transplantation, and thereafter the single doses of cyclosporine were often smaller than 3 mg/kg. Thus, the results suggest that cyclosporine displays dose-dependent, saturable pre-systemic metabolism even at low single doses, whereas complete saturation of CYP3A4 and MDR1 (P-glycoprotein) renders cyclosporine pharmacokinetics dose-linear at higher doses. No significant associations were found between genetic polymorphisms and cyclosporine pharmacokinetics before transplantation in the whole population for which genetic data was available (n=104). However, in children older than eight years (n=22), heterozygous and homozygous carriers of the ABCB1 c.2677T or c.1236T alleles had an about 1.3 times or 1.6 times higher oral bioavailability, respectively, than did non-carriers. After transplantation, none of the ABCB1 SNPs or any other SNPs were found to be associated with cyclosporine clearance or oral bioavailability in the whole population, in the patients older than eight years, or in the patients younger than eight years. In the whole population, in those patients carrying the NR1I2 g.-25385C–g.-24381A–g.-205_-200GAGAAG–g.7635G–g.8055C haplotype, however, the bioavailability of cyclosporine was about one tenth lower, per allele, than in non-carriers. This effect was significant also in a subgroup of patients older than eight years. Furthermore, in patients carrying the NR1I2 g.-25385C–g.-24381A–g.-205_-200GAGAAG–g.7635G–g.8055T haplotype, the bioavailability was almost one fifth higher, per allele, than in non-carriers. It may be possible to improve individualization of cyclosporine dosing in children by accounting for the effects of developmental factors (body weight, liver size), time after transplantation, and cyclosporine dosing frequency/formulation. Further studies are required on the predictive value of genotyping for individualization of cyclosporine dosing in children.
  • Kinnunen, Marja (2006)
    Past research has shown that the risk of cardiovascular disease is significantly lower among those with higher levels of physical activity. Hostility and its’ cognitive component, cynical distrust, have been found to associate inversely with several health behaviours. However, the results concerning the association between physical activity and hostility have remained ambiguous. Also, high-self-efficacy and positive outcome expectancies have been found to promote one’s physical activity. This cross-sectional population study (n=4957) examined if cynical distrust is related to leisure time physical activity. Also, it was explored if the associations of cynical distrust, self-efficacy on healthy lifestyle and outcome expectancies on healthy lifestyle together are associated with leisure time physical activity. In addition, the effects of age, body mass index (BMI), health status, education, and smoking were adjusted. Cynical distrust was associated inversely with leisure time physical activity (Beta = -0.107, p< 0 .001 in men, and Beta = -0.130, p< 0.001 in women). However, this association disappeared within men (Beta = -0.043, p=0.063), but not in women (Beta = -0.073, p< 0.001) after self-efficacy for healthy lifestyle and outcome expectancies for healthy lifestyle, age, and education were added into the model. The results of the present study suggest that leisure time physical activity is associated with different psychosocial determinants in men and women. Men’s leisure time physical activity is associated with self-efficacy for healthy lifestyle, outcome expectancies for healthy lifestyle, education, positive health status, lower BMI, and non-smoker status. Women’s leisure time physical activity was associated with lower levels of cynical distrust, self-efficacy for healthy lifestyle, higher outcome expectancies for healthy lifestyle, education, lower BMI, and non-smoker status.
  • Kinnunen, Marja (2006)
    Viimeaikaisten tutkimuksien tulokset osoittavat, että ihmisillä, jotka ovat fyysisesti aktiivisia on fyysisesti inaktiivisempia ihmisiä pienempi riski sairastua sydän- ja verisuonitauteihin. Vihamielisyydellä ja sen kognitiivisella komponentilla, kyynisellä epäluottamuksella, on todettu olevan negatiivisia yhteyksiä eri terveyskäyttäytymisiin. Kuitenkin tutkimustulokset koskien fyysisen aktiivisuuden ja vihamielisyyden välistä yhteyttä ovat edelleen epäselviä. Pystyvyyden tunteen ja positiivisten tulosodotuksien on todettu olevan merkitseviä selittäjiä liikuntakäyttäytymisessä. Tämä tutkimus on osa kansallista FINRISKI 2002 – tutkimusta, jossa seurataan sydän- ja verisuonitauteja. Tässä poikkileikkauksellissa väestötukimuksessa tarkasteltiin kyynisen epäluottamuksen yhteyttä vapaa-ajan fyysiseen aktiivisuuteen. Tutkimuksessa tarkasteltiin myös kyynisen epäluottamuksen, pystyvyyden tunteen sekä tulosodotusten yhteyttä vapaa-ajan fyysieen aktiivisuuteen. Tutkimuksessa huomioitiin myös osallistujien ikän, koulutus, painoindeksi, terveyden tila, koulutus sekä tupakointi. Tutkimukseen osallistui 4957 25–74 –vuotiasta miestä ja naista. Tutkimuksen tulos osoitti, että kyynisellä epäluottamuksella oli käänteinen yhteys vapaa-ajan fyysiseen aktiivisuuteen (Beta = -0.107, p< 0 .001miehillä, ja Beta= -0.130, p< 0.001 naisilla). Kyynisen epäluottamuksen ja vapaa-ajan fyysisen aktiivisuuden välinen todettu yhteys pysyi merkitsevänä naisilla (Beta = -0.073, p< 0.001), muttei kuitenkaan miehillä (Beta = -0.043, p=0.063), kun pystyvyys, tulosodotukset, ikä, koulutus, painoindeksi, terveyden tila sekä tupakointi lisättiin regressiomalliin. Tulokset osoittavat, että miesten ja naisten vapaa-ajan fyysisen aktiivisuuden taustalla on erilaisia psykososiaalisia tekijöitä. Miehet, jotka kokevat pystyvänsä elämään terveelisesti, omaavat positiiviset tulosodotukset terveellisiltä elämäntavoilta, ovat koulutetumpia, terveitä, omaavat pienemmän painoindeksin eivätkä tupakoi olivat todennäköisemmin fyysisesti aktiivisia. Naisilla tas korkeampaa fyysisen aktiivisuuden tasoa ennustivat matala kyynisen epäluottamuksen taso, pystyvyyden tunne elää terveellisesti, positiiviset tulosodotukset terveellisestä elämäntavasta, korkeampi koulutus, pienempi painoindeksi ja tupakoimattomuus.
  • Koskela, Outi (2012)
    Farmakogenetiikka on tieteenala, joka tutkii geneettisten erojen vaikusta lääkeaineiden vasteeseen eli farmakokinetiikkaan, tehoon ja haittavaikutuksiin. Kirjallisuuskatsaus käsittelee kipulääkkeiden farmakogenetiikkaa. Tutkituimmat kipulääkkeiden vastetta muuntavat geenivariaatiot ovat μ-opiodireseptoria koodaavan OPRM1-geenin 118A>Gmuutos sekä lääkeaineiden metaboliasta vastaavia sytokromi P450 (CYP) –entsyymejä koodaavien geenien polymorfiat. Myös P-glykoproteiinia koodaavan ABCB1-geenin ja COMT-geenin variaatioilla on osoitettu olevan vaikutusta kipulääkkeiden vasteeseen. Kokeellisessa osassa tutkittiin CYP2D6-, CYP3A4- ja CYP3A5-entsyymien geneettistä vaihtelua. Tavoitteena oli selvittää poikkeavatko CYP2D6-, CYP3A4- ja CYP3A5-geenien alleeli- ja haplotyyppifrekvenssit suomalaisten rintasyöpäpotilaiden ja terveiden vapaaehtoisten välillä. Jatkotutkimuksissa tullaan edelleen selvittämään miten näiden metaboliaentsyymien geneettinen vaihtelu vaikuttaa erään lääkeaineen vasteeseen. Tutkimukseen osallistui 996 suomalaista rintasyöpäpotilasta. Potilaiden DNA-näytteistä määritettiin CYP2D6-, 3A4- ja 3A5-geenien yleiset entsyymiaktiivisuuteen merkittävästi vaikuttavat variaatiot. CYP2D6-geenistä määritettiin 10 yksittäisen nukleotidin muutosta (SNP) sekä geenin kopioluku. CYP3A4-geenistä genotyypitettiin intronin 6 SNP rs35599367, jonka on osoitettu vähentävän CYP3A4-entsyymin ilmentymistä. Lisäksi genotyypitettiin CYP3A5-geenin 6986A>G SNP, joka johtaa lähetti-RNA:n silmikoitumisvirheeseen ja ennenaikaiseen lopetuskodoniin. Genotyypitykset ja kopiolukumääritykset suoritettiin polymeraasiketjureaktioon pohjautuvalla TaqMan® 5’-nukleaasimenetelmällä. CYP2D6-haplotyypit johdettiin genotyypitystuloksista tilastollisella analyysillä ja haplotyyppien perusteella muodostettiin potilaiden CYP2D6-fenotyyppiennuste. Ihmiset jaetaan CYP2D6-entsyymiaktiivisuuden mukaan neljään fenotyyppiluokkaan: hitaisiin (PM), keskinopeisiin (IM), nopeisiin (EM) ja erittäin nopeisiin (UM) metaboloijiin. Tutkimuspopulaatiossa CYP2D6 PM-fenotyyppi esiintyi 2,8 %:lla, IM-fenotyyppi 2,0 %:lla, EM-fenotyyppi 87,7 %:lla ja UM-fenotyyppi 7,6 %:lla potilaista. Tämän tutkimuksen CYP2D6 haplotyyppi- ja fenotyyppifrekvenssit ovat hyvin samanlaiset kuin aikaisemmin suomalaisille terveille vapaaehtoisille julkaistut frekvenssit. Tutkimuspopulaatiossa CYP3A4 rs35599367 SNP:n harvinaisemman alleelin frekvenssi oli 2,7 % ja CYP3A5:n 6986G>A SNP:n 7,6 %. Tutkimuksessa havaitut CYP3A5-alleelifrekvenssit vastaavat terveille vapaaehtoisille suomalaisille julkaistuja esiintymistiheyksiä. CYP3A4 rs35599367 SNP:n esiintymisestä suomalaisilla ei ole aikaisempaa julkaistua tutkimustietoa. Tutkimuksessa ei havaittu merkittäviä eroja CYP2D6- ja CYP3A5-geenien geneettisessä variaatiossa rintasyöpäpotilaiden ja terveiden vapaaehtoisten välillä. CYP3A4-geenin rs35599367 SNP:n esiintyminen suomalaisilla terveillä vapaaehtoisilla tulisi selvittää, jotta sitä voitaisiin verrata variaation esiintymiseen rintasyöpäpotilailla. Tämän tutkimuksen tulosten avulla voidaan edelleen selvittää CYP2D6-, CYP3A4- ja CYP3A5-entsyymien geneettisen vaihtelun vaikutusta lääkevasteeseen.
  • Lassus , Johan (Helsingin yliopisto, 2011)
    Acute heart failure (AHF) is a complex syndrome associated with exceptionally high mortality. Still, characteristics and prognostic factors of contemporary AHF patients have been inadequately studied. Kidney function has emerged as a very powerful prognostic risk factor in cardiovascular disease. This is believed to be the consequence of an interaction between the heart and kidneys, also termed the cardiorenal syndrome, the mechanisms of which are not fully understood. Renal insufficiency is common in heart failure and of particular interest for predicting outcome in AHF. Cystatin C (CysC) is a marker of glomerular filtration rate with properties making it a prospective alternative to the currently used measure creatinine for assessment of renal function. The aim of this thesis is to characterize a representative cohort of patients hospitalized for AHF and to identify risk factors for poor outcome in AHF. In particular, the role of CysC as a marker of renal function is evaluated, including examination of the value of CysC as a predictor of mortality in AHF. The FINN-AKVA (Finnish Acute Heart Failure) study is a national prospective multicenter study conducted to investigate the clinical presentation, aetiology and treatment of, as well as concomitant diseases and outcome in, AHF. Patients hospitalized for AHF were enrolled in the FINN-AKVA study, and mortality was followed for 12 months. The mean age of patients with AHF is 75 years and they frequently have both cardiovascular and non-cardiovascular co-morbidities. The mortality after hospitalization for AHF is high, rising to 27% by 12 months. The present study shows that renal dysfunction is very common in AHF. CysC detects impaired renal function in forty percent of patients. Renal function, measured by CysC, is one of the strongest predictors of mortality independently of other prognostic risk markers, such as age, gender, co-morbidities and systolic blood pressure on admission. Moreover, in patients with normal creatinine values, elevated CysC is associated with a marked increase in mortality. Acute kidney injury, defined as an increase in CysC within 48 hours of hospital admission, occurs in a significant proportion of patients and is associated with increased short- and mid-term mortality. The results suggest that CysC can be used for risk stratification in AHF. Markers of inflammation are elevated both in heart failure and in chronic kidney disease, and inflammation is one of the mechanisms thought to mediate heart-kidney interactions in the cardiorenal syndrome. Inflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) correlate very differently to markers of cardiac stress and renal function. In particular, TNF-α showed a robust correlation to CysC, but was not associated with levels of NT-proBNP, a marker of hemodynamic cardiac stress. Compared to CysC, the inflammatory markers were not strongly related to mortality in AHF. In conclusion, patients with AHF are elderly with multiple co-morbidities, and renal dysfunction is very common. CysC demonstrates good diagnostic properties both in identifying impaired renal function and acute kidney injury in patients with AHF. CysC, as a measure of renal function, is also a powerful prognostic marker in AHF. CysC shows promise as a marker for assessment of kidney function and risk stratification in patients hospitalized for AHF.
  • Isohanni, Mika (Helsingin yliopisto, 2009)
    Lidocaine is a widely used local anaesthetic agent that also has anti-arrhythmic effects. It is classified as a type Ib anti-arrhythmic agent and is used to treat ventricular tachycardia or ventricular fibrillation. Lidocaine is eliminated mainly by metabolism, and less than 5% is excreted unchanged in urine. Lidocaine is a drug with a medium to high extraction ratio, and its bioavailability is about 30%. Based on in vitro studies, the earlier understanding was that CYP3A4 is the major cytochrome P450 (CYP) enzyme involved in the metabolism of lidocaine. When this work was initiated, there was little human data on the effect of inhibitors of CYP enzymes on the pharmacokinetics of lidocaine. Because lidocaine has a low therapeutic index, medications that significantly inhibit lidocaine clearance (CL) could increase the risk of toxicity. These studies investigated the effects of some clinically important CYP1A2 and CYP3A4 inhibitors on the pharmacokinetics of lidocaine administered by different routes. All of the studies were randomized, double-blind, placebo-controlled cross-over studies in two or three phases in healthy volunteers. Pretreatment with clinically relevant doses of CYP3A4 inhibitors erythromycin and itraconazole or CYP1A2 inhibitors fluvoxamine and ciprofloxacin was followed by a single dose of lidocaine. Blood samples were collected to determine the pharmacokinetic parameters of lidocaine and its main metabolites monoethylglycinexylidide (MEGX) and 3-hydroxylidocaine (3-OH-lidocaine). Itraconazole and erythromycin had virtually no effect on the pharmacokinetics of intravenous lidocaine, but erythromycin slightly prolonged the elimination half-life (t½) of lidocaine (Study I). When lidocaine was taken orally, both erythromycin and itraconazole increased the peak concentration (Cmax) and the area under the concentration-time curve (AUC) of lidocaine by 40-70% (Study II). Compared with placebo and itraconazole, erythromycin increased the Cmax and the AUC of MEGX by 40-70% when lidocaine was given intravenously or orally (Studies I and II). The pharmacokinetics of inhaled lidocaine was unaffected by concomitant administration of itraconazole (Study III). Fluvoxamine reduced the CL of intravenous lidocaine by 41% and prolonged the t½ of lidocaine by 35%. The mean AUC of lidocaine increased 1.7-fold (Study IV). After oral administration of lidocaine, the mean AUC of lidocaine in-creased 3-fold and the Cmax 2.2-fold by fluvoxamine (Study V). During the pretreatment with fluvoxamine combined with erythromycin, the CL of intravenous lidocaine was 53% smaller than during placebo and 21% smaller than during fluvoxamine alone. The t½ of lidocaine was significantly longer during the combination phase than during the placebo or fluvoxamine phase. The mean AUC of intravenous lidocaine increased 2.3-fold and the Cmax 1.4-fold (Study IV). After oral administration of lidocaine, the mean AUC of lidocaine increased 3.6-fold and the Cmax 2.5-fold by concomitant fluvoxamine and erythromycin. The t½ of oral lidocaine was significantly longer during the combination phase than during the placebo (Study V). When lidocaine was given intravenously, the combination of fluvoxamine and erythromycin prolonged the t½ of MEGX by 59% (Study IV). Compared with placebo, ciprofloxacin increased the mean Cmax and AUC of intravenous lidocaine by 12% and 26%, respectively. The mean plasma CL of lidocaine was reduced by 22% and its t½ prolonged by 7% (Study VI). These studies clarify the principal role of CYP1A2 and suggest only a modest role of CYP3A4 in the elimination of lidocaine in vivo. The inhibition of CYP1A2 by fluvoxamine considerably reduces the elimination of lidocaine. Concomitant use of fluvoxamine and the CYP3A4 inhibitor erythromycin further increases lidocaine concentrations. The clinical implication of this work is that clinicians should be aware of the potentially increased toxicity of lidocaine when used together with inhibitors of CYP1A2 and particularly with the combination of drugs inhibiting both CYP1A2 and CYP3A4 enzymes.
  • Nieminen, Kaisa (Helsingin yliopisto, 2009)
    Secondary growth of plants is of pivotal importance in terrestrial ecosystems, providing a significant carbon sink in the form of wood. As plant biomass accumulation results largely from the cambial growth, it is surprising that quite little is known about the hormonal or genetic control of this important process in any plant species. The central aim of my thesis studies was to explore the function of cytokinin in the regulation of cambial development. Since their discovery as regulators of plant cell divisions, cytokinins have been assumed to participate in the control of cambial development. Evidence for this action was deduced from hormone treatment experiments, where exogenously applied cytokinin was shown to enhance cambial cell divisions in diverse plant organs and species. In my thesis work, the conservation of cytokinin signalling and homeostasis genes between a herbaceous plant, Arabidopsis, and a hardwood tree species, Populus trichocarpa. Presumably reflecting the ancient origin of cytokinin signalling system, the Populus genome contains orthologs for all Arabidopsis cytokinin signalling and homeostasis genes. Thus, genes belonging to five main families of isopentenyl transferases (IPTs), cytokinin oxidases (CKXs), two-component receptors, histidine containing phosphotransmitters (HPts) and response regulators (RRs) were identified from the Populus genome. Three subfamilies associated with cytokinin signal transduction, the CKI1-like family of two-component receptors, the AHP4-like HPts, and the ARR22-like atypical RRs, were significantly larger in Populus genome than in Arabidopsis. Potential contribution to the extensive secondary development of Populus by the members of these considerably expanded gene families will be discussed. Representatives of all cytokinin signal transduction elements were expressed in the Populus cambial zone, and most of the expressed genes appeared to be slightly more abundant on the phloem side of the meristem. The abundance of cytokinin related genes in the cambium emphasizes the important role of this hormone in the regulation of the extensive secondary growth characteristic of tree species. The function of the pseudo HPts in primary vascular development was studied in Arabidopsis root vasculature. It was demonstrated that the pseudo HPt AHP6 has a role in locally inhibiting cytokinin signalling in the protoxylem position in the Arabidopsis root, thus enabling differentiation of the protoxylem cell file. The possible role of pseudo HPts in cambial development will be discussed. The expression peak of cytokinin signalling genes in the tree cambial zone strongly indicates that cytokinin has a role in the regulation of this meristem function. To address whether cytokinin signalling is required for cambial activity, transgenic Populus trees with modified cytokinin signalling were produced. These trees were expressing a cytokinin catabolic gene from Arabidopsis, CYTOKININ OXIDASE 2, (AtCKX2) under the promoter of a Betula CYTOKININ RECEPTOR 1 (BpCRE1). The pBpCRE1::CKX2 transgenic Populus trees showed a reduced concentration of a biologically active cytokinin, correlating with their impaired cytokinin response. Furthermore, the radial growth of these trees was compromised, as illustrated by a smaller stem diameter than in wild-type trees of the same height. Moreover, the level of cambial cytokinin signalling was down-regulated in these thin-stemmed trees. The reduced signalling correlated with a decreased number of meristematic cambial cells, implicating cytokinin activity as a direct regulator of cambial cell division activity. Together, the results of my study indicate that cytokinins are major hormonal regulators required for cambial development.
  • Mähönen, Ari Pekka (Helsingin yliopisto, 2005)
  • Helanterä, Ilkka (Helsingin yliopisto, 2006)
    Cytomegalovirus (CMV) is a major cause of morbidity, costs and even mortality in organ transplant recipients. CMV may also enhance the development of chronic allograft nephropathy (CAN), which is the most important cause of graft loss after kidney transplantation. The evidence for the role of CMV in chronic allograft nephropathy is somewhat limited, and controversial results have also been reported. The aim of this study was to investigate the role of CMV in the pathogenesis of CAN. Material for the purpose of this study was available from altogether 70 kidney transplant recipients who received a kidney transplant between the years 1992-2000. CMV infection was diagnosed with pp65 antigenemia test or by viral culture from blood, urine, or both. CMV proteins were demonstrated in the kidney allograft biopsies by immunohistochemisrty and CMV-DNA by in situ hybridization. Cytokines, adhesion molecules, and growth factors were demonstrated from allograft biopsies by immunohistochemistry, and from urinary samples by ELISA-methods. CMV proteins were detectable in the 6-month protocol biopsies from 18/41 recipients with evidence of CMV infection. In the histopathological analysis of the 6-month protocol biopsies, presence of CMV in the allograft together with a previous history of acute rejection episodes was associated with increased arteriosclerotic changes in small arterioles. In urinary samples collected during CMV infection, excretion of TGF-β was significantly increased. In recipients with increased urinary excretion of TGF-β, increased interstitial fibrosis was recorded in the 6- month protocol biopsies. In biopsies taken after an active CMV infection, CMV persisted in the kidney allograft in 17/48 recipients, as CMV DNA or antigens were detected in the biopsies more than 2 months after the last positive finding in blood or urine. This persistence was associated with increased expression of TGF-β, PDGF, and ICAM-1 and with increased vascular changes in the allografts. Graft survival and graft function one and two years after transplantation were reduced in recipients with persistent intragraft CMV. Persistent intragraft CMV infection was also a risk factor for reduced graft survival in Cox regression analysis, and an independent risk factor for poor graft function one and two years after transplantation in logistic regression analysis. In conclusion, these results show that persistent intragraft CMV infection is detrimental to kidney allografts, causing increased expression of growth factors and increased vascular changes, leading to reduced graft function and survival. Effective prevention, diagnosis and treatment of CMV infections may a major factor in improving the long term survival of kidney allograft.
  • Uuksulainen, Pasi (2013)
    Tutkielma on rakenteeltaan kaksiosainen. Kirjallisuustutkimus on tehty Helsingin yliopistossa ja kokeellinen tutkimus Espanjassa (Universidad Complutense de Madrid). Tästä johtuen ensimmäinen osa on kirjoitettu suomeksi ja toinen osa englanniksi. Kirjallisuuskatsauksessa perehdytään C6-substituoitujen pyrimidiininukleosidien valmistukseen. C6-substituoiduilla pyrimidiininukleosideilla on havaittu olevan monia erilaisia biologisia vaikutuksia. C6-substituoiduista pyrmidiininukleosideista ei ole kovin paljon julkaisuja johtuen synteettisten keinojen vähäisyydestä. C6-substituoituja pyrimidiininukleosideja voidaan valmistaa joko yhdistämällä C6-substituoitu silyloitu emäs ja suojattu sokeri tai muokkaamalla suoraan valmiin nukleosidin C6-asemaa. Happamissa olosuhteissa glykosylaatioreaktiolla valmistettujen C6-substituoitujen pyrimidiininukleosidien saantoa voidaan parantaa käyttämällä Friedel-Crafts-katalyyttejä ja suojaamalla emäksen N3-asema. Glykosylaatioreaktiolla valmistettujen pyrimidiininukleosidien C6-asemassa olevat substituentit ovat yleensä pieniä ryhmiä. Valmiin pyrimidiininukleosidin C6-aseman muokkaukseen on useita keinoja, joista palladiumkatalyytit tarjoavat yhden tehokkaimmista keinoista liittää esimerkiksi aryyliryhmiä C6-asemaan. Muokkaamalla valmista nukleosidia vältetään myös glykosylaatioreaktiossa syntyvien N3-isomeerien muodostuminen. Joidenkin substituenttien kohdalla havaittiin N-glykosidisidoksen hajoamista. Kokeellisen tutkimuksen tavoitteena oli valmistaa uusia asymmetrisessä katalyysissä käytettäviä kiraalisia kamferijohdannaisia ligandeja. Kahden kamferijohdannaisen ligandin valmistuksessa saatiin valmistettua lupaavia välituotteita. Kolme polyhydroksyloitunutta norbornaani- johdannaista eivät toimineet kiraalisina ligandeina vaan tuottivat raseemisen seoksen.
  • D. 
    Puranen, Kaarina (1948)
  • Lehto, Susanna (2015)
    Dagumin jakauma on jatkuva todennäköisyysjakauma, joka on saanut nimensä Camilo Dagumin mukaan tämän esitellessä jakaumaa 1970-luvulla. Dagumin jakauman kehittäminen sai alkusysäyksen, kun Camilo Dagum ei ollut tyytyväinen jo olemassa oleviin todennäköisyysjakaumiin ja alkoi kehitellä vaatimuksiaan vastaavaa mallia. Tämän kehitystyön tuloksena syntyi kolme jakaumaa, joita kutsutaan Dagumin jakauman tyypeiksi I--III. Tyyppi I on kolme parametria sisältävä jakauma, kun taas tyypit II ja III ovat keskenään hyvin samankaltaisia, neljä parametria sisältäviä jakaumia. Dagumin jakauma tyypistä riippumatta kehitettiin kuvaamaan henkilökohtaisia tuloja, ja tämän vuoksi jakauma yhdistetään usein taloustieteen tulonjako-oppiin. Lisäksi Dagumin jakauman kolme tyyppiä voidaan luokitella tilastollisiksi kokojakaumiksi, joita usein hyödynnetään etenkin taloustieteessä ja vakuutusmatematiikassa. Luku 1 koostuu johdannosta, jossa esitellään pro gradu -tutkielman rakenne pääpiirteissään sekä valotetaan syitä, miksi juuri Dagumin jakauma valikoitui tutkielman aiheeksi. Luvussa 2 esitellään lyhyesti jatkuvien todennäköisyysjakaumien yleistä teoriaa siltä osin kuin sen tunteminen on vähintäänkin tarpeellista. Tässä yhteydessä esitellään myös tärkeitä merkintöjä erityisesti luvun 3 ymmärtämiseksi. Luku 3 alkaa Dagumin jakauman kehittäjän, Camilo Dagumin, henkilöhistorialla. Tästä päästään sujuvasti syihin, jotka motivoivat Dagumia entistä paremman mallin etsimiseen ja johtivat lopulta kokonaan uuden jakauman tai jakaumaperheen syntymiseen. Aivan tuulesta Dagumin jakaumaa ei kuitenkaan ole temmattu, vaan pohjalla on Dagumin laaja-alainen asiantuntemus ja useiden eri jakaumien ja mallien tutkiminen ja testaaminen. Vaikka Dagumin jakauma tyyppeineen on aivan oma jakaumansa, sillä on myös läheisiä yhteyksiä muihin jakaumiin ja näiden yhteyksien vuoksi siitä käytetään usein myös nimeä Burr III -jakauma. Luvussa 3 valotetaan lisäksi Dagumin jakauman perusominaisuuksia, joiden esittelyn myötä katse suunnataan jakauman käyttökelpoisuuteen sovelluksissa: jakauma osoittautuu hyödylliseksi tulonjaon tasa-arvoisuuden mittaamisessa, jossa myös estimoinnilla ja päätelmien tekemisellä on tärkeä rooli. Luvun lopussa käsitellään lyhyesti ja ytimekkäästi Dagumin jakauman käyttämistä tietokoneohjelmien avulla. Vaikka luvussa 3 viitataan monessa kohtaa Dagumin jakauman sovelluksiin, vasta luvussa 4 jakauman soveltaminen käytäntöön otetaan lähempään tarkasteluun. Viimeisessä luvussa kootaan päällimmäisiä ajatuksia ja mietteitä Dagumin jakaumasta sekä haasteista tutustua siihen: yhdessä pro gradussa pystytään vasta raapaisemaan pintaa, joten työsarkaa riittäisi muillekin jakaumasta kiinnostuneille.