Individualized immunosuppression after renal transplantation in childhood

Pediatric renal transplantation (TX) has evolved greatly during the past few decades, and today TX is considered the standard care for children with end-stage renal disease. In Finland, 191 children had received renal transplants by October 2007, and 42% of them have already reached adulthood. Improvements in treatment of end-stage renal disease, surgical techniques, intensive care medicine, and in immunosuppressive therapy have paved the way to the current highly successful outcomes of pediatric transplantation. In children, the transplanted graft should last for decades, and normal growth and development should be guaranteed. These objectives set considerable requirements in optimizing and fine-tuning the post-operative therapy. Careful optimization of immunosuppressive therapy is crucial in protecting the graft against rejection, but also in protecting the patient against adverse effects of the medication. In the present study, the results of a retrospective investigation into individualized dosing of immunosuppresive medication, based on pharmacokinetic profiles, therapeutic drug monitoring, graft function and histology studies, and glucocorticoid biological activity determinations, are reported. Subgroups of a total of 178 patients, who received renal transplants in 1988 2006 were included in the study. The mean age at TX was 6.5 years, and approximately 26% of the patients were <2 years of age. The most common diagnosis leading to renal TX was congenital nephrosis of the Finnish type (NPHS1). Pediatric patients in Finland receive standard triple immunosuppression consisting of cyclosporine A (CsA), methylprednisolone (MP) and azathioprine (AZA) after renal TX. Optimal dosing of these agents is important to prevent rejections and preserve graft function in one hand, and to avoid the potentially serious adverse effects on the other hand. CsA has a narrow therapeutic window and individually variable pharmacokinetics. Therapeutic monitoring of CsA is, therefore, mandatory. Traditionally, CsA monitoring has been based on pre-dose trough levels (C0), but recent pharmacokinetic and clinical studies have revealed that the immunosuppressive effect may be related to diurnal CsA exposure and blood CsA concentration 0-4 hours after dosing. The two-hour post-dose concentration (C2) has proved a reliable surrogate marker of CsA exposure. Individual starting doses of CsA were analyzed in 65 patients. A recommended dose based on a pre-TX pharmacokinetic study was calculated for each patient by the pre-TX protocol. The predicted dose was clearly higher in the youngest children than in the older ones (22.9±10.4 and 10.5±5.1 mg/kg/d in patients <2 and >8 years of age, respectively). The actually administered oral doses of CsA were collected for three weeks after TX and compared to the pharmacokinetically predicted dose. After the TX, dosing of CsA was adjusted according to clinical parameters and blood CsA trough concentration. The pharmacokinetically predicted dose and patient age were the two significant parameters explaining post-TX doses of CsA. Accordingly, young children received significantly higher oral doses of CsA than the older ones. The correlation to the actually administered doses after TX was best in those patients, who had a predicted dose clearly higher or lower (> ±25%) than the average in their age-group. Due to the great individual variation in pharmacokinetics standardized dosing of CsA (based on body mass or surface area) may not be adequate. Pre-Tx profiles are helpful in determining suitable initial CsA doses. CsA monitoring based on trough and C2 concentrations was analyzed in 47 patients, who received renal transplants in 2001 2006. C0, C2 and experienced acute rejections were collected during the post-TX hospitalization, and also three months after TX when the first protocol core biopsy was obtained. The patients who remained rejection free had slightly higher C2 concentrations, especially very early after TX. However, after the first two weeks also the trough level was higher in the rejection-free patients than in those with acute rejections. Three months after TX the trough level was higher in patients with normal histology than in those with rejection changes in the routine biopsy. Monitoring of both the trough level and C2 may thus be warranted to guarantee sufficient peak concentration and baseline immunosuppression on one hand and to avoid over-exposure on the other hand. Controlling of rejection in the early months after transplantation is crucial as it may contribute to the development of long-term allograft nephropathy. Recently, it has become evident that immunoactivation fulfilling the histological criteria of acute rejection is possible in a well functioning graft with no clinical sings or laboratory perturbations. The influence of treatment of subclinical rejection, diagnosed in 3-month protocol biopsy, to graft function and histology 18 months after TX was analyzed in 22 patients and compared to 35 historical control patients. The incidence of subclinical rejection at three months was 43%, and the patients received a standard rejection treatment (a course of increased MP) and/or increased baseline immunosuppression, depending on the severity of rejection and graft function. Glomerular filtration rate (GFR) at 18 months was significantly better in the patients who were screened and treated for subclinical rejection in comparison to the historical patients (86.7±22.5 vs. 67.9±31.9 ml/min/1.73m2, respectively). The improvement was most remarkable in the youngest (<2 years) age group (94.1±11.0 vs. 67.9±26.8 ml/min/1.73m2). Histological findings of chronic allograft nephropathy were also more common in the historical patients in the 18-month protocol biopsy. All pediatric renal TX patients receive MP as a part of the baseline immunosuppression. Although the maintenance dose of MP is very low in the majority of the patients, the well-known steroid-related adverse affects are not uncommon. It has been shown in a previous study in Finnish pediatric TX patients that steroid exposure, measured as area under concentration-time curve (AUC), rather than the dose correlates with the adverse effects. In the present study, MP AUC was measured in sixteen stable maintenance patients, and a correlation with excess weight gain during 12 months after TX as well as with height deficit was found. A novel bioassay measuring the activation of glucocorticoid receptor dependent transcription cascade was also employed to assess the biological effect of MP. Glucocorticoid bioactivity was found to be related to the adverse effects, although the relationship was not as apparent as that with serum MP concentration. The findings in this study support individualized monitoring and adjustment of immunosuppression based on pharmacokinetics, graft function and histology. Pharmacokinetic profiles are helpful in estimating drug exposure and thus identifying the patients who might be at risk for excessive or insufficient immunosuppression. Individualized doses and monitoring of blood concentrations should definitely be employed with CsA, but possibly also with steroids. As an alternative to complete steroid withdrawal, individualized dosing based on drug exposure monitoring might help in avoiding the adverse effects. Early screening and treatment of subclinical immunoactivation is beneficial as it improves the prospects of good long-term graft function.

Distal anterior cerebral artery aneurysms

Objective: Distal anterior cerebral artery (DACA) aneurysms represent about 6% of all intracranial aneurysms. So far, only small series on treatment of these aneurysms have been published. Our aim is to evaluate the anatomic features, microneurosurgical techniques, treatment results, and long-term outcome in patients treated for DACA aneurysms. Patients and methods: We analyzed the clinical and radiological data on 517 consecutive patients diagnosed with DACA aneurysm at two neurosurgical centers serving solely the Southern (Helsinki) and Eastern (Kuopio) Finland in 1936–2007, and used a defined subgroup of the whole study population in each part of the study. Detailed anatomic analysis was performed in 101 consecutive patients from 1998 to 2007. Treatment results were analyzed in 427 patients treated between 1980 to 2005, the era of CT imaging and microneurosurgery. Long-term treatment outcome of ruptured DACA aneurysm(s) was evaluated in 280 patients with a median follow-up of 10 years; no patients were lost to follow-up. Results: DACA aneurysms, found most often (83%) at the A3 segment of the anterior cerebral artery (ACA), were smaller (median 6 mm vs. 8 mm), more frequently associated with multiple aneurysms (35% vs. 18%), and presented more often with intracerebral hematomas (ICHs) (53% vs. 26%) than ruptured aneurysms in general. They were associated with anomalies of the ACA in 23% of the patients. Microsurgical treatment showed similar complication rates (treatment morbidity 15%, treatment mortality 0.4%) as for other ruptured aneurysms. At one year after subarachnoid hemorrhage (SAH), DACA aneurysms had equally favorable outcome (GOS≥4) as other ruptured aneurysms (74% vs. 69%) but their mortality was lower (13% vs. 24%). Factors predicting unfavorable outcome for ruptured DACA aneurysms were advanced age, Hunt&Hess≥3, rebleeding before treatment, ICH, intraventricular hemorrhage, and severe preoperative hydrocephalus. The cumulative relative survival ratio showed 16% excess mortality in patients with ruptured DACA aneurysm during the first three years after SAH compared to the matched general population. From the fourth year onwards, there was no excess mortality during the follow-up. There were four episodes of recurrent SAH, only one due to treated DACA aneurysm, with a 10-year cumulative risk of 1.4%. Conclusions: The special neurovascular features and frequent association with anterior cerebral artery anomalies must be taken into account when planning occlusive treatment of DACA aneurysms. Clipping of DACA aneurysms provides a long-lasting result, with very small rates of rebleeding. After surviving three years from rupture of DACA aneurysm, the long-term survival of these patients becomes similar to that of the matched general population.

Cyclosporine A in the treatment of Interstitial Cystitis

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.

Thrombin regulation in newborn infants

The coagulation system of newborn infants differs markedly from that of older children and adults. The activities of most coagulation factors and anticoagulants are low, leading to altered regulation in the formation of the key enzyme, thrombin. Timely and adequate generation of thrombin is essential, as thrombin activates platelets and many coagulation factors, cleaves fibrinogen into fibrin and activates the antithrombotic and anti-inflammatory protein C pathway. On the other hand, excess thrombin may promote thrombotic complications and exacerbate harmful inflammatory reactions. Despite the characteristic features, the newborn coagulation system can be considered physiological, since healthy newborns rarely show haemorrhagic or thrombotic complications. Sick newborns, however, often encounter clinical situations that challenge their coagulation system. The aim of this study was to clarify the behaviour of the neonatal coagulation system in selected clinical situations, with a special emphasis on the generation of thrombin. Thrombin was measured by in vivo thrombin generation markers and by thrombin generation potential in vitro. The patient groups included sick newborns undergoing intensive care and receiving fresh-frozen plasma (FFP), requiring exchange transfusions (ET) or presenting with a congenital heart defect requiring open heart surgery. Additionally, healthy newborns with inherited heterozygous factor V Leiden (FVL) mutation were studied. Thrombin generation potential was also analysed in cord plasma of healthy infants and in adults. Healthy as well as sick newborn infants showed lower total thrombin generation potential in vitro but faster initiation of thrombin generation than adults. These findings were qualitatively similar when plasma was supplemented with platelets. Platelets, however, significantly altered the effect of the major anticoagulant, activated protein C (APC), on thrombin generation potential. In accordance with previous studies, thrombin generation in healthy newborn platelet-poor plasma was resistant to the anticoagulant effects of APC, but when the plasma was supplemented with platelets APC attenuated thrombin generation significantly more in newborns than in adults. In vivo generation of thrombin was elevated in nearly all of the sick newborn infants. The low-volume FFP transfusion as opposed to the change from neonatal to adult blood in ET exerted markedly different effects on neonatal thrombin generation. FFP reduced the in vivo generation of thrombin in those newborns with the highest pretransfusional thrombin generation, thus acting as an anticoagulant agent. In those infants with lower pretransfusional thrombin generation, the effect of FFP on thrombin generation was fairly neutral. On the other hand, the combination of red blood cells and FFP, used to perform ET, significantly increased the in vivo thrombin formation and shifted the balance in the newborn coagulation system to the procoagulant direction. Cardiopulmonary bypass (CPB) also significantly increased the in vivo thrombin generation, but the thrombin generation profile during CPB differed from that previously observed in adults. Escalation of thrombin at early reperfusion was not observed in newborns; in adults, its occurrence is associated with postoperative myocardial damage. Finally, in healthy newborns with FVL heterozygosity, faster initiation of thrombin generation was observed compared with controls. Interestingly, FV level was lower in FVL-heterozygous infants, possibly to counteract the procoagulant effects induced by FVL. In conclusion, unique features regarding thrombin regulation in newborn infants were observed. These features included a novel platelet effect on the regulation of the protein C pathway. The clinical challenges mainly seemed to shift the balance in the coagulation system of newborns to the procoagulant direction. Blood component transfusions markedly affected coagulation in a manner specific to the product but that could also be altered by the clinical situation. Overall, the results highlight the need for understanding developmental haemostasis for both diagnostic and therapeutic purposes.

On the behaviour of some physical parameterization methods in high-resolution numerical weather prediction models

Modern-day weather forecasting is highly dependent on Numerical Weather Prediction (NWP) models as the main data source. The evolving state of the atmosphere with time can be numerically predicted by solving a set of hydrodynamic equations, if the initial state is known. However, such a modelling approach always contains approximations that by and large depend on the purpose of use and resolution of the models. Present-day NWP systems operate with horizontal model resolutions in the range from about 40 km to 10 km. Recently, the aim has been to reach operationally to scales of 1 4 km. This requires less approximations in the model equations, more complex treatment of physical processes and, furthermore, more computing power. This thesis concentrates on the physical parameterization methods used in high-resolution NWP models. The main emphasis is on the validation of the grid-size-dependent convection parameterization in the High Resolution Limited Area Model (HIRLAM) and on a comprehensive intercomparison of radiative-flux parameterizations. In addition, the problems related to wind prediction near the coastline are addressed with high-resolution meso-scale models. The grid-size-dependent convection parameterization is clearly beneficial for NWP models operating with a dense grid. Results show that the current convection scheme in HIRLAM is still applicable down to a 5.6 km grid size. However, with further improved model resolution, the tendency of the model to overestimate strong precipitation intensities increases in all the experiment runs. For the clear-sky longwave radiation parameterization, schemes used in NWP-models provide much better results in comparison with simple empirical schemes. On the other hand, for the shortwave part of the spectrum, the empirical schemes are more competitive for producing fairly accurate surface fluxes. Overall, even the complex radiation parameterization schemes used in NWP-models seem to be slightly too transparent for both long- and shortwave radiation in clear-sky conditions. For cloudy conditions, simple cloud correction functions are tested. In case of longwave radiation, the empirical cloud correction methods provide rather accurate results, whereas for shortwave radiation the benefit is only marginal. Idealised high-resolution two-dimensional meso-scale model experiments suggest that the reason for the observed formation of the afternoon low level jet (LLJ) over the Gulf of Finland is an inertial oscillation mechanism, when the large-scale flow is from the south-east or west directions. The LLJ is further enhanced by the sea-breeze circulation. A three-dimensional HIRLAM experiment, with a 7.7 km grid size, is able to generate a similar LLJ flow structure as suggested by the 2D-experiments and observations. It is also pointed out that improved model resolution does not necessary lead to better wind forecasts in the statistical sense. In nested systems, the quality of the large-scale host model is really important, especially if the inner meso-scale model domain is small.

Atomic scale engineering of carbon nanotubes

Carbon nanotubes, seamless cylinders made from carbon atoms, have outstanding characteristics: inherent nano-size, record-high Young’s modulus, high thermal stability and chemical inertness. They also have extraordinary electronic properties: in addition to extremely high conductance, they can be both metals and semiconductors without any external doping, just due to minute changes in the arrangements of atoms. As traditional silicon-based devices are reaching the level of miniaturisation where leakage currents become a problem, these properties make nanotubes a promising material for applications in nanoelectronics. However, several obstacles must be overcome for the development of nanotube-based nanoelectronics. One of them is the ability to modify locally the electronic structure of carbon nanotubes and create reliable interconnects between nanotubes and metal contacts which likely can be used for integration of the nanotubes in macroscopic electronic devices. In this thesis, the possibility of using ion and electron irradiation as a tool to introduce defects in nanotubes in a controllable manner and to achieve these goals is explored. Defects are known to modify the electronic properties of carbon nanotubes. Some defects are always present in pristine nanotubes, and naturally are introduced during irradiation. Obviously, their density can be controlled by irradiation dose. Since different types of defects have very different effects on the conductivity, knowledge of their abundance as induced by ion irradiation is central for controlling the conductivity. In this thesis, the response of single walled carbon nanotubes to ion irradiation is studied. It is shown that, indeed, by energy selective irradiation the conductance can be controlled. Not only the conductivity, but the local electronic structure of single walled carbon nanotubes can be changed by the defects. The presented studies show a variety of changes in the electronic structures of semiconducting single walled nanotubes, varying from individual new states in the band gap to changes in the band gap width. The extensive simulation results for various types of defect make it possible to unequivocally identify defects in single walled carbon nanotubes by combining electronic structure calculations and scanning tunneling spectroscopy, offering a reference data for a wide scientific community of researchers studying nanotubes with surface probe microscopy methods. In electronics applications, carbon nanotubes have to be interconnected to the macroscopic world via metal contacts. Interactions between the nanotubes and metal particles are also essential for nanotube synthesis, as single walled nanotubes are always grown from metal catalyst particles. In this thesis, both growth and creation of nanotube-metal nanoparticle interconnects driven by electron irradiation is studied. Surface curvature and the size of metal nanoparticles is demonstrated to determine the local carbon solubility in these particles. As for nanotube-metal contacts, previous experiments have proved the possibility to create junctions between carbon nanotubes and metal nanoparticles under irradiation in a transmission electron microscope. In this thesis, the microscopic mechanism of junction formation is studied by atomistic simulations carried out at various levels of sophistication. It is shown that structural defects created by the electron beam and efficient reconstruction of the nanotube atomic network, inherently related to the nanometer size and quasi-one dimensional structure of nanotubes, are the driving force for junction formation. Thus, the results of this thesis not only address practical aspects of irradiation-mediated engineering of nanosystems, but also contribute to our understanding of the behaviour of point defects in low-dimensional nanoscale materials.

Quantum Chemical Studies on Tropospheric Nucleation Mechanisms Involving Sulfuric Acid

Nucleation is the first step of the process by which gas molecules in the atmosphere condense to form liquid or solid particles. Despite the importance of atmospheric new-particle formation for both climate and health-related issues, little information exists on its precise molecular-level mechanisms. In this thesis, potential nucleation mechanisms involving sulfuric acid together with either water and ammonia or reactive biogenic molecules are studied using quantum chemical methods. Quantum chemistry calculations are based on the numerical solution of Schrödinger's equation for a system of atoms and electrons subject to various sets of approximations, the precise details of which give rise to a large number of model chemistries. A comparison of several different model chemistries indicates that the computational method must be chosen with care if accurate results for sulfuric acid - water - ammonia clusters are desired. Specifically, binding energies are incorrectly predicted by some popular density functionals, and vibrational anharmonicity must be accounted for if quantitatively reliable formation free energies are desired. The calculations reported in this thesis show that a combination of different high-level energy corrections and advanced thermochemical analysis can quantitatively replicate experimental results concerning the hydration of sulfuric acid. The role of ammonia in sulfuric acid - water nucleation was revealed by a series of calculations on molecular clusters of increasing size with respect to all three co-ordinates; sulfuric acid, water and ammonia. As indicated by experimental measurements, ammonia significantly assists the growth of clusters in the sulfuric acid - co-ordinate. The calculations presented in this thesis predict that in atmospheric conditions, this effect becomes important as the number of acid molecules increases from two to three. On the other hand, small molecular clusters are unlikely to contain more than one ammonia molecule per sulfuric acid. This implies that the average NH3:H2SO4 mole ratio of small molecular clusters in atmospheric conditions is likely to be between 1:3 and 1:1. Calculations on charged clusters confirm the experimental result that the HSO4- ion is much more strongly hydrated than neutral sulfuric acid. Preliminary calculations on HSO4- NH3 clusters indicate that ammonia is likely to play at most a minor role in ion-induced nucleation in the sulfuric acid - water system. Calculations of thermodynamic and kinetic parameters for the reaction of stabilized Criegee Intermediates with sulfuric acid demonstrate that quantum chemistry is a powerful tool for investigating chemically complicated nucleation mechanisms. The calculations indicate that if the biogenic Criegee Intermediates have sufficiently long lifetimes in atmospheric conditions, the studied reaction may be an important source of nucleation precursors.

Agoralla : avauksia ympäristöasiantuntijoiden vuorovaikutusprosesseista

Ympäristöasiantuntijoiden vuorovaikutusta on tutkittu agoralla (antiikin tori). Se on julkinen tila, jossa markkinat, politiikka, tiede ja yhteiskunta kohtaavat. Tutkimus kuuluu yhteiskuntatieteellisen ympäristötutkimuksen alaan, mutta siinä hyödynnetään myös tulevaisuudentutkimusta. Työn motivaationa on ollut tekijän monitieteinen koulutustausta: yhteiskuntatieteilijä ja luonnontieteilijä. Miten ja miksi vuorovaikutus eri asiantuntijoiden välillä on haasteellista ja merkityksellistä esimerkiksi metsän biodiversiteetin vähenemisen ehkäisemiseksi. Keskeisiä käsitteitä ovat asiantuntijuus, vuorovaikutus, tiedon luotettavuusja kontekstisidonnaisuus, Väitöskirja koostuu neljästä eri asiantuntijuustarinasta. Ensimmäinen (luku 2) perustuu haastatteluihin suomalaisten ja saksalaisten bio- ja yhteiskuntatietelijöiden käsityksistä luonnosta ja ympäristöstä. Tutkimusongelmana on luonnontieteilijöiden ja yhteiskuntatieteilijöiden Suomessa ja Saksassa ”kulttuurierot” luonnon ja ympäristön käsitteellistämisessä. Johtopäätöksenä on, että aistittu luonto, ympäröivä ympäristö sekä ihmisen muokkaama elinympäristö eivät tunne selkeitä tiede- eikä maanrajoja. Tämä luku toimii ponnahduslautana konstruktioiden taakse vuorovaikutuksen haasteisiin. Kirjan toinen tarina (luku 3) perustuu haastatteluihin suomalaisten metsän biodiversiteettiasiantuntijoiden vuorovaikutuksesta. Tutkimusongelman lähtökohtana on metsän biodiversiteetin väheneminen ja tästä seuraavat polittisetkin vuorovaikutustilanteet. Miten konteksti vaikuttaa eri asiantuntijoiden vuorovaikutukseen ja mitä tästä seuraa? Analyysin päätulos on implisiittisen, vahvasti kontekstisidonnaisen asiantuntijatiedon hyödyntämisen tarve ja voimavara metsän biodiversiteetin vähenemisen ennaltaehkäisemiksi. Kolmas tarina asiantuntijuudesta (luku 4) perustuu Etelä-Suomen metsien suojelutoimikunnassa (Metso) tehtyihin havainnointeihin. Tutkija on näin ollut itse eräänlaisella torilla havainnoijana. Tutkimusongelmana on ”ohipuhuminen”, tiedon luotettavuus ja implisiittien tiedon hyväksyttävyys. Johtopäätöksenä on asiantuntijuuden vahva kontekstisidonnaisuus hetkeen ja paikkaan ja yhteisen kielen (vrt. transdisiplinaarisuus) löytyminen yhteisen tavoitteen saavuttamiseksi. Merkittäviä välineitä vuorovaikutuksen onnistumiseen ovat esimerkiksi yhteinen vahva tavoitetila, interkatio, joka koskee läsnä olevia ihmisiä ei instituutioita sekä fasilitaattorin vahva rooli tulkkina ja välittäjänä. Neljäs tarina (luku 5) vie agoran konkretiaan. Tässä luvussa on kehitetty eläytymiskävely- menetelmää, jossa fasilitaattori (tutkija) johdattaa Espoon keskuksessa hallinnon, politiikan, asukkaiden ja konsultin edustajat aistimaan ja tulkitsemaan alueen sosiaalista tilaa, toiminnallisuutta ja elämyksellisyyttä. Ongelmana on aistimaailman asiantuntemuksen hyödyntämättömyys yhdyskuntasuunnittelun välineenä mm. asiantuntijoiden vuorovaikutuksen välineenä. Menetelmäkehitys on aluillaan, mutta jo tässä tapauksessa käy ilmi, että jaettu tila, jaetut aistikokemukset konkreettisella kävelyllä avaavaat vuorovaikutuksen uusiin ulottuvuuksiin, jossa implisiittiselle asiantuntemukselle annetaan sijansa vuorovaikutuksessa ja tätä kautta voidaan vaikuttaa myös tehtäviin päätöksiin, toimenpiteisiin. Johtopäätöksissä (luku 6) korostuu implisiittisen asiantuntijuuden merkitys. Onnistunut vuorovaikutteinen toiminta eri asiantuntijoiden kesken esimerkiksi erilaisia ympäristöongelmia –ja ilmiöitä ratkottaessa ja pohdittaessa vaatii vuorovaikutusosaamista. Tutkimuksen lopuksi suositellaan esimerkiksi ennakkoluulottomia avauksia agoralla. Asiantuntijuus ei ole yksi ja vain asiaatuntevuus on mahdollista. Agora on jatkuvassa liikkeessä ja juuri siinä piilee voimavara tulevaisuuden haasteisiin erilaisilla rajapinnoilla. Avainsanat: asiantuntijuus, vuorovaikutus, tieto, konteksti, agora

Vapaaehtoiset vuodeosastolla : Etnografinen tutkimus vanhusten ja vapaaehtoisten kohtaamisesta

This ethnographic study investigates encounters between volunteers and older people at the Kerava Municipal Health Centre inpatient ward for chronic care. Volunteer activities have been under development, in cooperation with the Voluntary Work Center (Talkoorengas), since the start of the 1990s. When my research began in 2003, nine of the volunteers came to the ward on set days per week or visited the ward according to their own timetables. The volunteers ranged in age from 54 to 78 years. With one exception, all of them were on pension. Nearly all of them had been volunteers for more than ten years. My study is research on ageing, the focal point being older people, whether volunteers or those receiving assistance. The research questions are: How is volunteer work implemented in daily routines at the ward? How is interaction created in encounters between the older people and the volunteers? What meanings does volunteer work create for the older people and the volunteers? The core material of my research is observation material, which is supplemented by interviews, documentation and photographs. The materials have been analysed by using theme analysis and ethnomethodological discussion analysis. In the presentation of the research findings, I have structured the materials into three main chapters: space and time; hands and touch; and words and tones. The chapter on space and time examines time and space paths, privacy and publicness, and celebrations as part of daily life. The volunteers open and create social arenas for the older people through chatting and singing together, celebrations in the dayroom or poetry readings at the bedside. The supporting theme of the chapter on hands and touch is bodily closeness in care and the associated concrete physical presence. The chapter highlights the importance of everyday routines, such as meals and rituals, as elements that bring security. Stimuli in daily life, such as handicrafts in groups, pass time but also give older people the experience of meaningful activity and bring back positive memories of their own life. The chapter on words and tones focuses on the social interaction and identity. The volunteers’ identity is built up into the identity of a helper and caregiver. The older people’s identity is built up into a care recipient’s identity, which in different situations is shaped into, among others, the identity of one who listens, remembers, does not remember, defends, composes poetry or is dying. The cornerstones of voluntary social care are participation, activity, trust and presence. Successful volunteer work calls for mutual trust between the older people, volunteers and the health care personnel, and for clear agreements on questions of responsibility, the status of volunteers and their role alongside professional personnel. This study indicates that volunteer work is a meaningful resource in work with older people.

Spectroscopy of tissue triglyceride composition : In vitro and in vivo studies

Lipid analysis is commonly performed by gas chromatography (GC) in laboratory conditions. Spectroscopic techniques, however, are non-destructive and can be implemented noninvasively in vivo. Excess fat (triglycerides) in visceral adipose tissue and liver is known predispose to metabolic abnormalities, collectively known as the metabolic syndrome. Insulin resistance is the likely cause with diets high in saturated fat known to impair insulin sensitivity. Tissue triglyceride composition has been used as marker of dietary intake but it can also be influenced by tissue specific handling of fatty acids. Recent studies have shown that adipocyte insulin sensitivity correlates positively with their saturated fat content, contradicting the common view of dietary effects. A better understanding of factors affecting tissue triglyceride composition is needed to provide further insights into tissue function in lipid metabolism. In this thesis two spectroscopic techniques were developed for in vitro and in vivo analysis of tissue triglyceride composition. In vitro studies (Study I) used infrared spectroscopy (FTIR), a fast and cost effective analytical technique well suited for multivariate analysis. Infrared spectra are characterized by peak overlap leading to poorly resolved absorbances and limited analytical performance. In vivo studies (Studies II, III and IV) used proton magnetic resonance spectroscopy (1H-MRS), an established non-invasive clinical method for measuring metabolites in vivo. 1H-MRS has been limited in its ability to analyze triglyceride composition due to poorly resolved resonances. Using an attenuated total reflection accessory, we were able to obtain pure triglyceride infrared spectra from adipose tissue biopsies. Using multivariate curve resolution (MCR), we were able to resolve the overlapping double bond absorbances of monounsaturated fat and polyunsaturated fat. MCR also resolved the isolated trans double bond and conjugated linoleic acids from an overlapping background absorbance. Using oil phantoms to study the effects of different fatty acid compositions on the echo time behaviour of triglycerides, it was concluded that the use of long echo times improved peak separation with T2 weighting having a negligible impact. It was also discovered that the echo time behaviour of the methyl resonance of omega-3 fats differed from other fats due to characteristic J-coupling. This novel insight could be used to detect omega-3 fats in human adipose tissue in vivo at very long echo times (TE = 470 and 540 ms). A comparison of 1H-MRS of adipose tissue in vivo and GC of adipose tissue biopsies in humans showed that long TE spectra resulted in improved peak fitting and better correlations with GC data. The study also showed that calculation of fatty acid fractions from 1H-MRS data is unreliable and should not be used. Omega-3 fatty acid content derived from long TE in vivo spectra (TE = 540 ms) correlated with total omega-3 fatty acid concentration measured by GC. The long TE protocol used for adipose tissue studies was subsequently extended to the analysis of liver fat composition. Respiratory triggering and long TE resulted in spectra with the olefinic and tissue water resonances resolved. Conversion of the derived unsaturation to double bond content per fatty acid showed that the results were in accordance with previously published gas chromatography data on liver fat composition. In patients with metabolic syndrome, liver fat was found to be more saturated than subcutaneous or visceral adipose tissue. The higher saturation observed in liver fat may be a result of a higher rate of de-novo-lipogenesis in liver than in adipose tissue. This thesis has introduced the first non-invasive method for determining adipose tissue omega-3 fatty acid content in humans in vivo. The methods introduced here have also shown that liver fat is more saturated than adipose tissue fat.

Two ex situ fungal technologies to treat contaminated soil

Wood-degrading fungi are able to degrade a large range of recalcitrant pollutants which resemble the lignin biopolymer. This ability is attributed to the production of lignin-modifying enzymes, which are extracellular and non-specific. Despite the potential of fungi in bioremediation, there is still an understanding gap in terms of the technology. In this thesis, the feasibility of two ex situ fungal bioremediation methods to treat contaminated soil was evaluated. Treatment of polycyclic aromatic hydrocarbons (PAHs)-contaminated marsh soil was studied in a stirred slurry-phase reactor. Due to the salt content in marsh soil, fungi were screened for their halotolerance, and the white-rot fungi Lentinus tigrinus, Irpex lacteus and Bjerkandera adusta were selected for further studies. These fungi degraded 40 - 60% of a PAH mixture (phenanthrene, fluoranthene, pyrene and chrysene) in a slurry-phase reactor (100 ml) during 30 days of incubation. Thereafter, B. adusta was selected to scale-up and optimize the process in a 5 L reactor. Maximum degradation of dibenzothiophene (93%), fluoranthene (82%), pyrene (81%) and chrysene (83%) was achieved with the free mycelium inoculum of the highest initial biomass (2.2 g/l). In autoclaved soil, MnP was the most important enzyme involved in PAH degradation. In non-sterile soil, endogenous soil microbes together with B. adusta also degraded the PAHs extensively, suggesting a synergic action between soil microbes and the fungus. A fungal solid-phase cultivation method to pretreat contaminated sawmill soil with high organic matter content was developed to enhance the effectiveness of the subsequent soil combustion. In a preliminary screening of 146 fungal strains, 28 out of 52 fungi, which extensively colonized non-sterile contaminated soil, were litter-decomposing fungi. The 18 strains further selected were characterized by their production of lignin-modifying and hydrolytic enzymes, of which MnP and endo-1,4-β-glucanase were the main enzymes during cultivation on Scots pine (Pinus sylvestris) bark. Of the six fungi selected for further tests, Gymnopilus luteofolius, Phanerochaete velutina, and Stropharia rugosoannulata were the most active soil organic matter degraders. The results showed that a six-month pretreatment of sawmill soil would result in a 3.5 - 9.5% loss of organic matter, depending on the fungus applied. The pretreatment process was scaled-up for a 0.56 m3 reactor, in which perforated plastic tubes filled with S. rugosoannulata growing on pine bark were introduced into the soil. The fungal pretreatment resulted in a soil mass loss of 30.5 kg, which represents 10% of the original soil mass (308 kg). Despite the fact that Scots pine bark contains several antimicrobial compounds, it was a suitable substrate for fungal growth and promoter of the production of oxidative enzymes, as well as an excellent and cheap natural carrier of fungal mycelium. This thesis successfully developed two novel fungal ex situ bioremediation technologies and introduce new insights for their further full-scale application. Ex situ slurry-phase fungal reactors might be applied in cases when the soil has a high water content or when the contaminant bioavailability is low; for example, in wastewater treatment plants to remove pharmaceutical residues. Fungal solid-phase bioremediation is a promising remediation technology to ex situ or in situ treat contaminated soil.

Paluu paratiisiin : Polkuja hyvän ja pahan tuolle puolen Martti Lutherin ja Friedrich Nietzschen perinteissä

Tutkimukseni tarkoitus on paikallistaa polkuja hyvän ja pahan tuolle puolen. Tutkimukseni termit hyvä ja paha määrittyvät moraalisesti. Täten tutkimusaiheeni on ihmisen ja moraalin välinen suhde. Tutkin ihmisen moraalista olotilaa, siinä tapahtuvaa muutosta ja tämän muutoksen ehtoja. Tutkimuksellani on kaksi pääkysymystä: kuinka on mahdollista päästä hyvän ja pahan tuolle puolen ja miksi meidän tulisi siirtyä hyvän ja pahan tuolle puolen. Polut hyvän ja pahan tuolle puolen vievät myös etiikan tuolle puolen. Tutkimukseni pyrkii selvittämään mitä vikaa etiikassa on ja kuinka on mahdollista perustella toimintaa ilman etiikkaa. Tutkimukseni metodi on systemaattinen analyysi. Johdanto-osassa käsittelen yleisesti moraalia, sen vaikutuspiiriä ja niitä syitä, joiden vuoksi se täytyy hylätä. Tarkastelen myös etiikkaa ja selvitän sitä laajempaa kontekstia mihin se liittyy. Ensimmäisessä luvussa käsittelen Martti Lutherin teologiaa. Lutherilla moraalia edustaa Jumalan laki, jonka alaisuudesta kristitty pääsee vapaaksi kun Jeesus ottaa ristinkuolemassaan ihmisten synnin itselleen ja kärsii Jumalan lain mukaisen kuolemanrangaistuksen. Uskon yhteydessä Kristukseen kristitty kohoaa lain ulkopuolelle tekemään rakkauden tekoja. Toinen luku on omistettu Søren Kierkegaardin kristilliselle eksistentialismille. Kierkegaardin eksistenssitasojen filosofiassa uskonnollinen olemisen taso paljastuu korkeammaksi kuin eettinen. Tässä nousee tärkeäksi Kierkegaardin kirja Pelko ja vavistus. Kierkegaard tutkii Raamatun kertomusta Aabrahamista, jolle Jumala antaa käskyn uhrata oma poikansa Iisak. Kierkegaardille tarina on esimerkki eettistä korkeammasta ja sitä tilapäisesti vastustavasta olemisen tavasta. Kolmannessa luvussa käsittelen Dietrich Bonhoefferin teologiaa. Bonhoefferin mukaan Jeesus kutsuu ihmisiä seuraamaan häntä. Tämä on kutsu hyvän ja pahan tuolle puolen paratiisin viattomuuteen, joka rikkoo eettisyyden ihmisen ja Jeesuksen välillä. Neljännessä luvussa tarkastelen Friedrich Nietzschen filosofiaa ja pyrin ymmärtämään mitä hänen ohjelmansa hyvän ja pahan tuolle puolen tarkoittaa. Nietzschen vallantahdon filosofiassa moraali on heikkojen keino alistaa vahvat. Tarkastelen Nietzschen käsitystä moraalin synnystä ja sen haitallisuudesta ja pyrin selvittämään kuinka herrat elävät hyvän ja pahan tuolla puolen. Viidennessä luvussa tutkin Jaques Derridan grammatologiaa. Derridan mukaan teksti on suljettu tila, jonka ulkopuolelle on mahdoton päästä. Derrida sanoittaa kuitenkin sanoittamatonta tilattomuutta tekstin tuolla puolen. Tästä ei-paikasta käsin hypereettinen dekonstruktio purkaa perinteistä etiikkaa. Kuudennessa luvussa tarkastelen Mark C. Taylorin epä/teologiaa. Taylorille olotilan muutosta kuvaa termi harhautuminen , joka vie ihmisen lainsuojattomuuteen hyvän ja pahan tuolle puolen. Selvitän mitä harhautuminen tarkoittaa ja miten se liittyy Taylorin Jumalan kuoleman epä/teologiaan. Johtopäätösluvussa tuon esiin tutkimustuloksia, nostan esille joitain pääluvuissa esiintyviä teemoja ja vertaan tutkimuskohteitteni näkemyksiä toisiinsa. Lutherin ja Nietzschen perinteissä on paljon yhteistä, mutta pohjimmiltaan ne sotivat toisiaan vastaan. Hyvän ja pahan tuolle puolen johtaa kaksi polkua. Toinen niistä vie paratiisiin, toinen tuhoon.

Expression of lactate transporters MCT1, MCT2, MCT4 and the ancillary protein CD147 in horse muscle and red blood cells

Monocarboxylate transporters (MCTs) transport lactate and protons across cell membranes. During intense exercise, lactate and protons accumulate in the exercising muscle and are transported to the plasma. In the horse, MCTs are responsible for the majority of lactate and proton removal from exercising muscle, and are therefore also the main mechanism to hinder the decline in pH in muscle cells. Two isoforms, MCT1 and MCT4, which need an ancillary protein CD147, are expressed in equine muscle. In the horse, as in other species, MCT1 is predominantly expressed in oxidative fibres, where its likely role is to transport lactate into the fibre to be used as a fuel at rest and during light work, and to remove lactate during intensive exercise when anaerobic energy production is needed. The expression of CD147 follows the fibre type distribution of MCT1. These proteins were detected in both the cytoplasm and sarcolemma of muscle cells in the horse breeds studied: Standardbred and Coldblood trotters. In humans, training increases the expression of both MCT1 and MCT4. In this study, the proportion of oxidative fibres in the muscle of Norwegian-Swedish Coldblood trotters increased with training. Simultaneously, the expression of MCT1 and CD147, measured immunohistochemically, seemed to increase more in the cytoplasm of oxidative fibres than in the fast fibre type IIB. Horse MCT4 antibody failed to work in immunohistochemistry. In the future, a quantitative method should be introduced to examine the effect of training on muscle MCT expression in the horse. Lactate can be taken up from plasma by red blood cells (RBCs). In horses, two isoforms, MCT1 and MCT2, and the ancillary protein CD147 are expressed in RBC membranes. The horse is the only species studied in which RBCs have been found to express MCT2, and the physiological role of this protein in RBCs is unknown. The majority of horses express all three proteins, but 10-20% of horses express little or no MCT1 or CD147. This leads to large interindividual variation in the capacity to transport lactate into RBCs. Here, the expression level of MCT1 and CD147 was bimodally distributed in three studied horse breeds: Finnhorse, Standardbred and Thoroughbred. The level of MCT2 expression was distributed unimodally. The expression level of lactate transporters could not be linked to performance markers in Thoroughbred racehorses. In the future, better performance indexes should be developed to better enable the assessment of whether the level of MCT expression affects athletic performance. In human subjects, several mutations in MCT1 have been shown to cause decreased lactate transport activity in muscle and signs of myopathy. In the horse, two amino acid sequence variations, one of which was novel, were detected in MCT1 (V432I and K457Q). The mutations found in horses were in different areas compared to mutations found in humans. One mutation (M125V) was detected in CD147. The mutations found could not be linked with exercise-induced myopathy. MCT4 cDNA was sequenced for the first time in the horse, but no mutations could be detected in this protein.

Pylväskäytävien suojassa : Pompejilainen peristyylipuutarha sosioekonomisen edustamisen muotona

Pro gradu -tutkielmani käsittelee pompejilaisessa yksityistalossa ollutta puutarhaa, jota kiertää pylväskäytävä tai useampia. Tätä tilaa kutsutaan peristyylipuutarhaksi. Peristyyli oli tärkeä osa roomalaisen ja siis myös pompejilaisen talon julkista osaa. Se oli avoin vierailijoille, siksi voidaan olettaa, että peristyylipuutarhalla olisi haluttu esittää omaa vaurautta ja sosiaalista asemaa. Työni keskittyy siihen, miten talon asukkaan varallisuus ja sosiaalinen asema näkyvät peristyylipuutarhassa. Tutkin myös, mitä muuta peristyyli viestii omistajastaan. Tutkimukseni liitteenä on lista Pompejin yksityistalojen peristyyleistä ja niiden tiedoista. Niitä vertailemalla analysoin sitä, millainen tila peristyylipuutarha on ollut. Aineistona tutkielmassani ovat konkreettisesti Pompejin yksityistalojen peristyylipuutarhat ja niiden rakenteet sekä arkeologiset löydöt. Peristyylipuutarhoja on 168 kappaletta. Lisäksi seinämaalaukset ja piirtokirjoitukset välittävät tietoa niin peristyylistä kuin sen omistajasta. Tiedot peristyyleistä olen kerännyt tutkimuskirjallisuudesta löytyneistä luetteloista, joissa on tietoa Pompejin taloista ja puutarhoista. Murto-osassa peristyylipuutarhoista olen käynyt ja kerännyt niistä tietoa työhöni. Huomioin antiikin kirjallisen tradition, sen käsitellessä aihettani. Ensimmäisenä käsittelen peristyylipuutarhaa arkkitehtuurin kannalta. Tutkin, mihin peristyyli on sijoitettu pompejilaisessa talossa ja millaisia arkkitehtonisia ratkaisuja tilassa on käytetty. Peristyyli on usein talon ovelta avautuvalla näköakselilla, joka leikkaa läpi talon tärkeimpien julkisten tilojen. Ne ovat atrium eli eteishalli, tablinum eli vastaanottohuone ja peristyylipuutarha. Aina peristyyli ei ole tällä akselilla. Joissain taloissa peristyyli voi olla atriumin tai tablinumin paikalla. Pompejin peristyyleissä on pylväskäytäviä yhdestä neljään. Usein puutarhaosa on erotettu pylväskäytävistä aidalla. Seuraavaksi tutkin peristyylipuutarhasta löytyneitä rakenteita, irtaimistoa, kasveja ja istutuksia. Tarkastelen näiden yleisyyttä ja sitä, mitä ne kertovat peristyylin käyttötarkoituksesta. Puutarhoissa on ollut vesikouruja, kaivoja, altaita, suihkulähteitä, vesisäiliöitä ja niiden suuaukkojen renkaita. Kiinteitä rakennelmia ovat tricliniumit eli oleskelu- ja ruokailuryhmät, kotijumalien pyhäköt, alttarit, nissit ja katokset. Irtaimia löytöjä on peristyyleissä monenlaisia. Yleisimpiä ovat pöydät, aurinkokellot, lamput, amforat ja erilaiset astiat sekä ruukut. Kasveja on huomioitu vähän Pompejin kaivauksissa, mutta peristyylipuutarhoista on löytynyt merkkejä puista ja pensaista sekä säännöllisistä ja epäsäännöllisistä istutuksista. Viimeisenä käsittelen peristyylipuutarhan koristelua, ja minkälaisen kuvan se välittää talon omistajasta. Puutarhoissa oli veistoksia ja seinämaalauksia. Molemmissa käsitellään luontoaiheita, jotka ovat helposti liitettävissä puutarhaan. Suosittuja ovat myös mytologiset aiheet. Veistoskoristelussa selkeästi yleisin on viinin jumala Dionysos ja häneen liittyvät hahmot. Myös muita jumaluuksia esiintyy kuten Venus. Se ja Dionysos liitettiin antiikissa luontoon ja puutarhaan. Peristyylipuutarha oli monipuolinen tila, jossa eri elämänalueet kohtasivat toisensa. Peristyyleissä voitiin työskennellä. Puutarhaosa saattoi olla hyötykäytössä esimerkiksi kasvimaana. Peristyylissä tai sen ympäristössä oleskeltiin, juhlittiin ja viihdytettiin vieraita, siksi siellä esitettiin omaa varakkuutta runsaalla veistoskoristelulla, altailla ja suihkulähteillä. Jo laaja pinta-alainen peristyyli oli osoitus vauraudesta. Aina ei haluttu näyttää varakkuutta, vaan joskus koristeellinen peristyyli oli syrjässä talon ovelta avautuvalta näköakselilta. Uskonto oli osa peristyyliä, vaikka uskonnon harjoittamiseen liittyvät rakenteet sijoitettiin yleensä talon muihin tiloihin. Uskonto ja mytologia ovat kuitenkin vahvasti läsnä peristyylien koristelussa. Selkeitä poliittisia viittauksia peristyylipuutarhoissa on vähän, mutta roomalaisia piirteitä niissä voidaan havaita. Roomalaiset vaikutteet peristyylin koristelussa ja arkkitehtuurissa viestittävät mahdollisesti talon omistajan roomalaismielisyydestä. Roomalaiset piirteet ovat myös olleet trendejä aikakauden peristyyleissä.