TURUN KAUPUNGIN YMPÄRISTÖNSUOJELUTOIMISTON PROJEKTIT 1995–2000 Yhteiskunnallisen vaikuttavuuden arviointi

Tässä pro gradu -tutkielmassa arvioin Turun kaupungin ympäristönsuojelutoimiston vuosina 1995–2000 toteuttamien projektien yhteiskunnallista vaikuttavuutta. Projektit ovat: muovijätteen kierrättämiseen keskittynyt KAPULA, ilmanlaadun seurannan kehittämiseen luotu City Simulator, eri sektorien väliseen ympäristöyhteistyöhön kannustanut Green Know-How Turku, Hirvensalon kaavoituksen tulevaisuutta valottanut ICTULA sekä kansainvälinen sähköautoprojekti EVD-POST. Kaikki viisi projektia on toteutettu yhteistyönä, ja osallistujina niissä on ollut ympäristönsuojelutoimiston lisäksi niin Turun alueen kuntia, yrityksiä kuin eurooppalaisia kaupunkejakin. Tutkielmassa arvioin projektien yhteiskunnallista vaikuttavuutta julkishallinnollisesta näkökulmasta. Tässä tapauksessa yhteiskunnallinen vaikuttavuus merkitsee muun muassa sitä, miten ympäristönsuojelutoimisto on vastannut sille asetettuihin tavoitteisiin ja miten toiminta on vastannut siihen kohdistuvia yhteiskunnallisia tarpeita. Tutkimukseni perusteella tarpeet ovat muodostuneet 1990-luvun Suomen talouslaman, 1980- ja 1990-lukujen globaalin ja EU:n ympäristöpolitiikan sekä 1990-luvun kaupunkistrategioiden ja ympäristönsuojeluohjelman kautta. Projektien arviointiin hyödynnän tavoitelähtöistä metodia Evert Vedungin Public Policy and Program Evaluation sekä Huey-Tsyh Chenin Theory-Driven Evaluations -teoksia. Metodilla tarkennan tavoitteiden, toimenpiteiden ja lopputulemien suhdetta. Sen lisäksi käytän Ray Pawsonin ja Nick Tilleyn Realistic Evaluation -kirjan realistista arviointitapaa, jolla tutkin kontekstin ja tehtyjen toimenpiteiden vaikutusta projekteissa. Projektien arvioinnin jälkeen kokoan yhteen päätelmiä projektien yhteiskunnallisesta vaikuttavuudesta ja pohdin syitä siihen, miksi projektit eroavat vaikuttavuudeltaan. Tutkielman johtopäätöksinä on, että projektit eroavat yhteiskunnalliselta vaikuttavuudeltaan lähtökohtien ja hyväksyttävyyden johdosta esimerkiksi lainsäädännön ja ympäristöpolitiikan vuoksi. Kyse on osaksi myös siitä, että joissain projekteissa tavoitteet on asetettu epärealistisiksi suhteessa niiden toteutumiseen. Projekteihin kohdistuvat ulkoiset tarpeet, toimintaympäristö ja realistiset tavoitteet eivät kuitenkaan ehdottomasti takaa yhteiskunnallista vaikuttavuutta. Myös yhteistyöprojektien toteuttajien ominaisuudet eli sisäiset tekijät vaikuttavat yhteiskunnallisen vaikuttavuuden ilmenemiseen.

Pissakoulu lasten toiminnallisen kastelun hoidossa: Intervention vaikuttavuuden arviointi kastelun määrän ja lasten kokemuksien avulla

Kastelu on merkittävästi lapsen arkielämää haittaava kiusallinen oire. Koulun aloittavista perusterveistä lapsista 10 – 25 % kastelee viikoittain joko päivällä ja/tai yöllä. Keskeistä kastelevan lapsen hoidossa on kastelua vähentävien ja virtsarakon hallintaa lisäävien tapa-asioiden ohjaus ja uusien toimintamallien vakiinnuttaminen lapsen arkeen. International Children’s Continence Societyn (ICCS) suositusten mukaan arjen tapa-asioiden ohjaukseen perustuvan uroterapian tulisi olla kastelevan lapsen ensisijainen hoitomuoto. Tutkimuksen tarkoituksena oli arvioida pissakoulu-intervention vaikuttavuutta kastelun hoidossa sekä kuvata lasten kokemuksia pissakoulusta. Tutkimusasetelmassa käytettiin toisiaan täydentävinä sekä kvantatiivisia että kvalitatiivisia aineistonkeruumenetelmiä. Kuusikymmentäyhdeksän hoitoresistentiksi kastelijaksi luokiteltua 6-12-vuotiasta lasta osallistui polikliiniseen pissakouluun 4-6 lapsen ryhmissä kahtena erillisenä päivänä kahden kuukauden välein. Otoskooksi määriteltiin voima-analyysin perusteella 52. Ohjauksen sisältö perustui ICCS:n suosituksiin ja toteutuksessa painotettiin lapsilähtöisyyttä sekä yhdessä tekemistä ja keskustelua. Kvantitatiivisen osan aineistonkeruumenetelmänä käytettiin viikon WC-päiväkirjaa. Kastelun määrän muutoksia mitattiin ennen ja jälkeen molempien pissakoulupäivien ja tulokset analysoitiin SPSS tilasto-ohjelmalla. Kvalitatiivinen aineisto kerättiin pissakoulun toisen päivän päätteeksi haastattelemalla viisi ensimmäistä pissakouluryhmää, jonka jälkeen aineiston todettiin saturoituneen. Ryhmäkeskustelut analysoitiin aineistolähtöisellä sisällönanalyysillä. Tutkimukseen osallistuneista 69 lapsesta 58:lta saatiin tiedot kaikilta neljältä mittauskerralta. Lapset olivat iältään keskimäärin 8,3 vuotiaita, 34 oli tyttöä ja 24 poikaa. Verrattaessa ensimmäisen ja viimeisen mittauksen keskiarvojen välisiä eroja kuivat päivät (3,5 vrt. 5,3, p<0.001) ja kuivat yöt (2,4 vrt. 3,9, p<0.001) lisääntyivät tilastollisesti merkitsevästi. Päiväkastelun osalta 24/40 (60 %) ja yökastelun osalta 23/46 (50 %) lapsella kastelu väheni ≥50 %. Kuivaksi tuli 13/58 (22 %) lasta. Ryhmähaastatteluihin osallistui 19 lasta (12 tyttöä ja 7 poikaa, keski-ikä 8,9 vuotta). Lapset kokivat kastelun monella tapaa arkea haittaavaksi noloksi pulmaksi. Pissakoulu auttoi heitä tulemaan pissarakon pomoksi, jolloin pulmasta tuli hallittava tai siitä pääsi kokonaan eroon. Erityisen tärkeää lapsille oli tavata toisia kastelevia lapsia ja jakaa kokemuksia heidän kanssaan sekä oppia tekemällä lapsilähtöisin menetelmin. Pissakoulu on lapsilähtöinen ja vaikuttava hoitotyön interventio, joka arjen tapa-asioiden muutoksilla auttaa lapsia saavuttamaan paremman virtsarakon hallinnan ja vähentämään kastelua tarjoten myös arvokasta vertaistukea.

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Stem Cell Transplantation for Retinal Degenerations

Within the last few years, several reports have revealed that cell transplantation can be an effective way to replace lost neurons in the central nervous system (CNS) of patients affected with neurodegenerative diseases. Concerning the retina, the concept that newborn photoreceptors can integrate the retina and restore some visual functions was univocally demonstrated recently in the mouse eye (MacLaren et al. 2006) and remains to be achieved in human. These results pave the way to a standard approach in regenerative medicine aiming to replace lost photoreceptors. With the discovery of stem cells a great hope has appeared towards elaborating protocols to generate adequate cells to restore visual function in different retinal degeneration processes. Retinal stem cells (RSCs) are good candidates to repair the retina and are present throughout the retina development, including adulthood. However, neonatal mouse RSCs derived from the radial glia population have a different potential to proliferate and differentiate in comparison to adult RSCs. Moreover, we observed that adult mouse RSCs, depending on the culture conditions, have a marked tendency to transform, whereas neonatal RSCs show subtle chromosome abnormalities only after extensive expansion. These characteristics should help to identify the optimal cell source and culture conditions for cell transplantation studies. These results will be discussed in light of other studies using RSCs as well as embryonic stem cells. Another important factor to consider is the host environment, which plays a crucial role for cell integration and which was poorly studied in the normal and the diseased retina. Nonetheless, important results were recently generated to reconsider cell transplantation strategy. Perspectives to enhance cell integration by manipulating the environment will also be presented.

Octopus Standard Automated Perimetry, Pulsar Perimetry, Moorfields Motion Displacement Test and Heidelberg Retinal Tomography in Glaucoma Detection - A Comparison of Diagnostic Accuracy

Purpose: To investigate the accuracy of 4 clinical instruments in the detection of glaucomatous damage. Methods: 102 eyes of 55 test subjects (Age mean = 66.5yrs, range = [39; 89]) underwent Heidelberg Retinal Tomography (HRTIII), (disc area<2.43); and standard automated perimetry (SAP) using Octopus (Dynamic); Pulsar (TOP); and Moorfields Motion Displacement Test (MDT) (ESTA strategy). Eyes were separated into three groups 1) Healthy (H): IOP<21mmHg and healthy discs (clinical examination), 39 subjects, 78 eyes; 2) Glaucoma suspect (GS): Suspicious discs (clinical examination), 12 subjects, 15 eyes; 3) Glaucoma (G): progressive structural or functional loss, 14 subjects, 20 eyes. Clinical diagnostic precision was examined using the cut-off associated with the p<5% normative limit of MD (Octopus/Pulsar), PTD (MDT) and MRA (HRT) analysis. The sensitivity, specificity and accuracy were calculated for each instrument. Results: See table Conclusions: Despite the advantage of defining glaucoma suspects using clinical optic disc examination, the HRT did not yield significantly higher accuracy than functional measures. HRT, MDT and Octopus SAP yielded higher accuracy than Pulsar perimetry, although results did not reach statistical significance. Further studies are required to investigate the structure-function correlations between these instruments.

Autofluorescence Findings Of A Novel Maculopathy In Patient With Spinocerebellar Ataxia Type 1 And Of A Cone - Rod Type Retinal Dysfunction In Three Generation Family With Spinocerebellar Ataxia Type 7

Purpose: To report a novel maculopathy in a patient with SCA1. To describe autofluorescence findings in family with SCA7 and associated cone-rod retinal dysfunction.Methods: 4 affected patients from two families were assessed to investigate a progressive loss of visual acuity (VA). Examinations included fundus photography, autofluorescence (AF) fundus fluorescein angiogragraphy (FFA) and optical coherence tomography. Electroretinogram (full-field) was performed in 2 affected patients. All patients had color vision testing using Ishihara pseudoisochromatic plates. Molecular analysis was performed in family 2.Results: The patient with known diagnosis of SCA1 had a visual acuity of 20/200 bilaterally and dyschromatopsia. He had saccadic pursuit. Fundus examination showed mild retinal pigment epithelium (RPE) changes at the macula. OCT showed bilateral macular serous detachment, which was not obvious at the FFA and explained his VA. AF imaging showed a central hyperfluorescence. The 45 year old proband from family 2 had a visual acuity of 200/20 and dyschromatopsia. ERG testing showed cone type dysfunction of photoreceptors. Her daughter affected at a younger age had the same ERGs findings. Fundus examination showed mild RPE changes in proband, normal findings in her daughter. AF imaging of both patients showed a ring of high density AF around the fovea. The ring was also obvious on near infrared AF. Later onset of gait imbalance led to the diagnosis of SCA7Conclusions: Within the group of spinocerebellar ataxias, only the type 7 is associated with retinal dysfunction. We present the first report of maculopathy associated with SCA1 causing severe vision loss. The ring of high density AF in SCA7 confirmed an early retinal photoreceptor dysfunction in patient with normal fundus.

Mfge8 Expression In Conjunctival Melanocytic Proliferations

Purpose: Milk fat globule epidermal growth factor-8 (MFGE8) is a secreted phosphatidylserine-binding protein that has been involved in phagocytosis, as well as in VEGF dependent neovascularization. In a study evaluating protein expression in membrane rafts of cutaneous melanoma at different stages of progression, MFGE8 expression was only identified in membrane rafts of metastatic cutaneous melanoma cell lines. Furthermore, MFGE8, identified at higher level in the vertical growth phase of cutaneous melanoma, promoted tumor growth in vivo, enhanced invasion in vitro and metastatic spread in a mouse model. The purpose of this study was to assess the expression of MFGE8 in conjunctival melanocytic proliferations.Methods: MFGE8 expression was assessed by immunohistochemistry in 66 melanocytic conjunctival proliferations including 21 conjunctival naevi, 20 Primary Acquired Melanosis (PAM) including (4 PAM without atypia and 16 PAM with atypia) and 25 conjunctival melanomas. Expression was independently assessed by 2 pathologists. Relevant clinico-pathological data were retrieved. Statistical anaylis was performed using JUMP 8 software.Results: The concordance between the 2 pathologists had an 87,5% agreement on the first independent assessment of MFGE8 expression. Complete agreement was further reached after joint revision of discordant cases. In the naevi, MFGE8 expression was found in only 4 cases (3 subepithelial cases and 1 composed combined naevus). In the PAM group, MFGE8 was identified in 1 PAM without atypia and 10 PAM with atypia. In the melanoma group, MFGE8 expression was observed in 68% of cases. The expression of MFGE8 in the conjunctival melanocytic proliferation was significantly higher in the melanoma (p=0,0009) and in the PAM (p=0,0169) than in naevi. Within the PAM subgroup, we found no significant correlation between MFGE8 expression and the presence of atypia in the respective specimen examined so far.Conclusions: We demonstrate a significant higher expression of MFGE8 in conjunctival melanoma compared to benign melanocytic lesions, suggesting that this protein may play a role in tumor progression of conjunctival melanocytic proliferations. Further experimental studies should be performed to better characterize MFGE8 involvement in conjunctival melanoma tumorigenesis.

Bilateral uveal pigmented alterations with remarkable evolution: long term follow-up in a case of possible bilateral diffuse uveal melanocytic proliferation

Purpose: to describe a case of probable bilateral diffuse uveal melanocytic proliferation (BDUMP) with scleral involvement, free from systemic malignancies and cataract. Methods: fifty months of follow up with recurrent complete ophthalmological examinations, including fundus photography, fluorescein/indocyanine green angiography (FA) and optical coherence tomography (OCT). Investigations also included an electroretinography (ERG) and histological examination of scleral biopsy. Extraocular malignancies were repeatedly searched. Results: the patient was a 61 year-old Italian man with chronic hepatitis type C. At first visit his best corrected visual acuity (BCVA) was 20/32 in OS and 20/25 in OD. Funduscopy showed multiple patch-shaped pigmented alterations involving macular region and mid retinal periphery. FA showed corresponding areas of late-phase hyperfluorescent pinpoints (figure 1a, OS) and intemediate-phase hypocyanescence (figure 1b, OS), with subtle serous neurosensory retinal detachment confirmed by OCT. Photopic and scotopic ERG tested normal. Systemic prednisone was administered for one month without any improvement. After ten months round pigmentary lesions appeared also in superior scleral surface of both eyes. Biopsy allowed to disclose slightly pigmented spindle cells. BCVA worsened for further 10 months, with enlargement of FA alteration areas but lenses still clear. After 30 months spontaneous coalescence and atrophy of retinal lesions started, paralleled by progressive visual recovery. At the end of our follow up BCVA was 20/25 in OU while scleral pigmentary lesions remained unchanged. Conclusions: we report the case of a patient with main features of BDUMP and some unusual findings. Although not all classical diagnostic criteria were fulfilled, the presence of scleral pigmented lesions and spontaneous visual recovery may enlarge clinical spectrum of the disease.

Variable phenotypic expressivity in a Swiss family with autosomal dominant retinitis pigmentosa due to mutation T494m in the Prpf3 gene

Purpose:to describe the clinical features in a five generations family segregating autosomal dominant retinitis pigmentosa and to identify the causative gene Patient and Methods:Twenty five individuals of a large five-generation family originating from Western Switzerland were ascertained for phenotypic and genotypic characterization. Ophthalmologic evaluations included color vision testing, Goldman perimetry and digital fundus photography. Some patients had autofluorescence (AF) imaging, ocular coherence tomography (OCT) and ISCEV-standard full-field electroretinography (ERG). Blood samples were collected from 10 affected (4 to 70 years of age) and 15 unaffected members after informed consent. DNA was isolated and exons and intron-exons junctions of known adRP genes were sequenced using a Big Dye sequencing kit 1.1. Results:Age of onset of nightblindness and severity of progression of the disease was variable between members of the family. Some patients had early onset of nightblindess aged 3, others at mid-twenties. Most patients had visual acuity above 0.6 for the first 4 decades. Two older patients still had good vision (0.4) in their seventies. Myopia (range: -2 to -5) was noticed in most affected subjects. Fundus findings showed areas of atrophy along the arcades. The AF imaging showed a large high density ring bilaterally. A T494M change was found in exon 11 of PRPF3 gene. The change segregates with the disease in the family. Conclusion: A mutation in the PRPF3 gene is rare compared with other genes causing ADRP. Although a T494M change has been reported, our family is the first one with a variable expressivity. Mutations in PRPF3 gene can cause a variable phenotype of ADRP unlike the previously described Danish and English families. Our report gives a better understanding as to the phenotype/genotype description of ADRP due to PRPF3 mutation.

Three Generation Family With Pattern Dystrophy and a Successful Treatment of Subfoveal Cnv Related to Pattern Dystrophy With Anti-vegf Intravitreal Injections

Purpose: To phenotype a large 3 generation Swiss family with pattern dystrophy and to report a successful result of treatment with ranibizumab of a subfoveal choroidal neovascularisation (CNV) associated with pattern dystrophy in 1 patient Patients and methods: 4 affected and 3 unaffected patients (3 female 4 male, age range: 19 - 80 years) were assessed with a complete ophthalmologic examination. AF images were taken using Heidelberg Retina Angiograph and the digital color photos, fluorescein angiogragraphy (FFA) using the same TOPCON 501 camera. Electroretinogram (full-field and multifocal) was performed in 1 affected patient. One 48 years old patient developed a subfoveal CNV, which was treated with 2 injections of ranibizumab, at 3 months interval. Blood sample was taken for molecular analysis (screening of the gene RDS). Results: Two patients had a typical fundoscopic appearance of pattern dystrophy with butterfly shaped deposit at the fovea and some peripheral flecks, as shown with AF imaging.. Two others affected patients had a more unusual appearance with some macular atrophy in one or both eyes, surrounded by flecks. The visual acuity ranged from 1.0 to 0.1 according to Snellen EDTRS chart. The patient with subfoveal CNV presented a drop of vision form 1.0 to 0.6 within 10 days prior to the diagnosis and also reported some metamorphopsia. FFA and optical computerized tomography (OCT) confirmed a classic CNV. After the 1st injection her vision improved to 1.0 but persistent metamorphopsia and fluid on OCT motivated a second injection. One month after the second injection the OCT was flat and the patient had no symptoms. The results of RDS screening will be presented at the meeting. Conclusion: We present a family with pattern dystrophy, with some members having an unusual fundus appearance, which was mistaken for an early onset dry AMD. The AF imaging is a useful tool in diagnosing this condition. A CNV associated with pattern dystrophy a rare. This is the first report of a successful treatment of the CNV with anti-VEGF intravitreal injections.

Cell Adhesion Peptide RGDSP and Laminin Support Proliferation and Migration of Postnatal Retinal Stem Cells in a 3D Hydrogel Culture System

Purpose: Retinal stem cells (RSCs) can be isolated from radial glia population of the newborn mouse retina (Angénieux et al., 2006). These RSCs have great capacity to renew and generate neurons including cells differentiated towards the photoreceptor lineage (Mehri-Soussi et al., 2006). However, our published results showed poor integration and survival rate after cell grafting into the retina. The uncontrollable environment of retina seems to be the problem. To bypass this, we are trying to generate hemi-retinal tissue in vitro that can be used for transplantation. Methods: Expanded RSCs were seeded in a mixture of poly-ethylene-glycol (PEG)-polymer-based hydrogels crosslinked by peptides that also serve as substrates for matrix metalloproteinases. Different doses of crosslinker peptides were tested. Several growth factors were studied to stimulate cell proliferation and differentiation. Results: Cells were trapped in hydrogels and cultured in the presence of FGF2 and EGF. Spherical cell clusters indicating proliferation appeared within several days, but there was no cell migration within the gel. We then added cell adhesion molecules integrin ligand RGDSP, or laminin, or a combination of both, into the gel. Cells grown with laminin showed the best proliferation. Cells grown with RGDSP proliferated a few times and then started to spread out. Cells grown with the combination of RGDSP and laminin showed better proliferation than with RGDSP alone and larger spread-outs than with laminin alone. After stimulations with first FGF2 and EGF, and then only FGF2, some cells showed neuronal morphology after 2 weeks. The neuronal population was assessed by the presence of neuronal marker b-tubulin-III. Glial cells were also present. Further characterizations are undergoing. Conclusions: RSC can grow and migrate in 3D hydrogel with the addition of FGF2, EGF, RGDSP and laminin. Further developments are necessary to form a homogenous tissue containing retinal cells.

Transcriptional Activation and Receptor Dimerization is Affected by Mutations in the NR2E3 Ligand-Binding Domain

Purpose: Mutations in the ligand-binding domain (LBD) of NR2E3 cause recessively inherited enhanced short wavelength sensitive (S-) cone syndrome (ESCS), Goldmann-Favre syndrome (GFS) and clumped pigmentary retinal degeneration (CPRD). In addition to ligand binding, the LBD contains also essential amino acid sequences for the oligomerization of nuclear receptors. The aim of our studies is to characterize the impact of mutations in the LBD on receptor oligomerization and transcriptional activity of NR2E3. Methods: The different NR2E3 mutants were generated by QuickChange mutagenesis and analyzed in 293T-based transactivation studies and BRET2 (bioluminescence resonance electron transfer) assays. In silico homology modeling of mutant proteins was also performed using available crystallographic data of related nuclear receptors. Results: The mutants p.W234S, p.A256V, p.A256E, p.L263P, p.R309G, p.R311Q, p.R334G, p.L336P, p.L353V, p.R385P and p.M407K, all located in the LBD, showed impaired receptor dimerization at various degrees. Impaired repressor dimerization as assessed by BRET2 assays did not always correlate with impaired repressor function of NR2E3 as assessed by cell-based reporter assays. There were minor differences of transcriptional activity of mutant proteins on mouse S-opsin (opn1sw), mouse cone arrestin (arr3) and human cone arrestin, suggesting that the effect of LBD mutations was independent of the promoter context. Conclusions: Mutational analysis and homology modeling allowed the characterization of potential oligomerization interfaces of the NR2E3 LBD. Additionally, mutations in NR2E3 LBD may cause recessive retinal degenerations by different molecular mechanisms.

. Phenotype/genotype correlation associated with a novel nonsense Glu20stop mutation in the Rp2 gene in a three-generation family with x-linked retinitis pigmentosa

Purpose: To assess the phenotype of patients in a large 3 generation Swiss family with X-linked retinitis pigmentosa (XLRP) due to a novel nonsense mutation Glu20stop in RP2 gene and to correlate with the genotype. Methods: 6 affected patients (1 male, 5 females, age range: 23 - 73 years) were assessed with a complete ophthalmologic examination. All had fundus autofluorescence images, standardised electroretinography, Goldmann visual fields and Optical Coherence Tomography. In addition, medical records of 2 affected male patients were reviewed. Blood sample was taken for molecular analysis. Results: The male patients were severely affected at a young age with early macular involvement. The youngest 23 y old male had also high myopia and vision of less than 0.05 according to Snellen EDTRS chart bilaterally. All 5 female carriers had some degree of rod-cone dystrophy, but no macular involvement. The visual acuity was 1.0 in the younger carriers, while the 73 years old had VA of 0.5. Two females had mild myopia (range -0.75 to -2) and one had anisometropia of 3.5D, with the more severely affected eye being myopic. Three out of 5 female carriers had optic nerve drusen. Conclusions: We report a novel Glu20stop mutation in RP2 gene, which is a rare cause of XLRP. Our description of severe phenotype in male patients with high myopia and early macular atrophy confirms previous reports. Unlike previous reports, all our female carriers had RP, but not macular involvement or high myopia. The identifiable phenotype for RP2-XLRP aids in clinical diagnosis and targeted genetic screening.