Seeing smells: development of an optoelectronic nose

The development of an array of chemically-responsive dyes on a porous membrane and in its use as a general sensor for odors and volatile organic compounds (VOCs) is reviewed. These colorimetric sensor arrays (CSA) act as an "optoelectronic nose" by using an array of multiple dyes whose color changes are based on the full range of intermolecular interactions. The CSA is digitally imaged before and after exposure and the resulting difference map provides a digital fingerprint for any VOC or mixture of odorants. The result is an enormous increase in discriminatory power among odorants compared to prior electronic nose technologies. For the detection of biologically important analytes, including amines, carboxylic acids, and thiols, high sensitivities (ppbv) have been demonstrated. The array is essentially non-responsive to changes in humidity due to the hydrophobicity of the dyes and membrane.

Tacrolimus vs. Azathioprine in myasthenia gravis treatment: a multicenter, prospective, randomized, double blind, head to head clinical trial

Current evidences show that tacrolimus could be a good therapeutic option in patients non-responsive to first-line treatments and even it could become one of the first-line drugs for treating MG. Our study will try to determine the effectivity and safety of tacrolimus in 190 myasthenia gravis patients with suboptimal response to azathioprine: after 12 months of treatment they will be randomized to continue with azathioprine or they will be switched to tacrolimus. Patients will follow the allocated treatment during 14 months and the response will be assessed as well as adverse effectsPURPOSE: To compare azathioprine vs. tacrolimus effectivity and safety in MG patients with suboptimal response to azathioprine at 12 monthsDESIGN: A 14 months multicenter, prospective, randomized, double blind, head to head clinical trial

Saako historiasta selvää? Monikulttuuriset yläkoululaiset historian lähteillä historian taidot motiivien, seurauksien, historian tulkintojen ja lähteiden luotettavuuden arvioinneissa

The research assesses the skills of upper comprehensive school pupils in history. The focus is on locating personal motives, assessing wider reasons hidden in historical sources and evaluating source reliability. The research also questions how a wide use of multiple sources affects pupils’ holistic understanding of historical phenomena. The participants were a multicultural group of pupils. The origins of their cultures can be traced to the Balkan, the Middle East, Asia and Europe. The number of native Finnish speakers and pupils speaking Finnish as their second language was almost equal. The multicultural composition provides opportunities to assess how culturally responsive learning history from sources is. The intercultural approach to learning in a multicultural setting emphasizes equality as a precondition for learning. In order to set assignments at least to some extent match with all participants only those answers were taken into account which were produced by pupils who had studied history for a similar period of time in the Finnish comprehensive school system. Due to the small number of participants (41), the study avoids wide generalizations. Nevertheless, possible cultural blueprints in pupils’ way of thinking are noted. The first test examined the skills of pupils to find motives for emigration. The results showed that for 7th graders finding reasons is not a problematic task. However, the number of reasons noticed and justifications varied. In addition, the way the pupils explained their choices was a distinguishing factor. Some pupils interpreted source material making use of previous knowledge on the issue, while other pupils based their analysis solely on the text handed and did not try to add their own knowledge. Answers were divided into three categories: historical, explanatory and stating. Historical answers combined smoothly previously learned historical knowledge to one’s own source analysis; explanatory answers often ignored a wider frame, although they were effective when explaining e.g. historical concepts. The stating answers only noticed motives from the sources and made no attempts to explain them historically. Was the first test culturally responsive? All pupils representing different cultures tackled the first source exam successfully, but there were some signs of how historical concepts are understood in a slightly different way if the pupil’s personal history has no linkage to the concepts under scrutiny. The second test focused on the history of Native Americans. The test first required pupils to recognize whether short source extracts (5) were written by Indians or Caucasians. Based on what they had already learned from North American history, the pupils did not find it hard to distinguish between the sources. The analysis of multiphase causes and consequences of the disputes between Native Americans and white Americans caused dispersion among pupils. Using two historical sources and combining historical knowledge from both of them simultaneously was cumbersome for many. The explanations of consequences can be divided into two groups: the ones emphasizing short term consequences and those placing emphasis on long term consequences. The short term approach was mainly followed by boys in every group. The girls mainly paid attention to long term consequences. The result suggests that historical knowledge in sources is at least to some extent read through role and gender lenses. The third test required pupils to explain in their own words how the three sources given differed in their account of living conditions in Nazi Germany, which turned out to be demanding for many pupils. The pupils’ stronghold was rather the assessment of source reliability and accounts why the sources approached the same events differently. All participants wrote critical and justified comments on reliability and aspects that might have affected the content of the sources. The pupils felt that the main reasons that affected source reliability were the authors’ ethnic background, nationality and profession. The assessment showed that pupils were well aware that position in a historical situation has an impact on historical accounts, but in certain cases the victim’s account was seen as a historical truth. The account of events by a historian was chosen most often as the most reliable source, but it was often justified leniently with an indication to professionalism rather than with clear ideas of how historians conduct accounts based on sources. In brief, the last source test demonstrates that pupils have a strong idea that the ethnicity or nationalism determines how people explained events of the past. It is also an implication that pupils understand how historical knowledge is interpretative. The results also imply that history can be analyzed from a neutral perspective. One’s own membership in an ethnical or religious group does not automatically mean that a person’s cultural identity excludes historical explanations if something in them contradicts with his or her identity. The second method of extracting knowledge of pupils’ historical thinking was an essay analysis. The analysis shows that an analytical account of complicated political issues, which often include a great number of complicated political concepts, leads more likely to an inconsistent structure in the written work of pupils. The material also demonstrates that pupils have a strong tendency to take a critical stance when assessing history. Historical empathy in particular is shown if history somehow has a linkage to young people, children or minorities. Some topics can also awake strong feelings, especially among pupils with emigrant background, if there is a linkage between one’s own personal history and that of the school; and occasionally a student’s historical experience or thoughts replaced school history. Using sources during history lessons at school seems to have many advantages. It enhances the reasoning skills of pupils and their skills to assess the nature of historical knowledge. Thus one of the main aims and a great benefit of source work is to encourage pupils to express their own ideas and opinions. To conclude, when assessing the skills of adolescents in history - their work with sources, comments on history, historical knowledge and finally their historical thinking - one should be cautious and avoid cut off score evaluations. One purpose of pursuing history with sources is to encourage pupils to think independently, which is a useful tool for further identity construction. The idea that pupils have the right to conduct their own interpretations of history can be partially understood as part of a wider learning process, justification to study history comes from extrinsic reasons. The intrinsic reason is history itself; in order to understand history one should have a basic understanding of history as a specific domain of knowledge. Using sources does not mean that knowing history is of secondary importance. Only a balance between knowing the contextual history, understanding basic key concepts and working with sources is a solid base to improve pupils’ historical understanding.

Targeting striatal metabotropic glutamate receptor type 5 in Parkinson's disease: bridging molecular studies and clinical trials

Metabotropic glutamate (mGlu) receptors are G protein-coupled receptors expressed primarily on neurons and glial cells modulating the effects of glutamatergic neurotransmission. The pharmacological manipulation of these receptors has been postulated to be valuable in the management of some neurological disorders. Accordingly, the targeting of mGlu5 receptors as a therapeutic approach for Parkinson's disease (PD) has been proposed, especially to manage the adverse symptoms associated to chronic treatment with classical PD drugs. Thus, the specific pharmacological blocking of mGlu5 receptors constitutes one of the most attractive non-dopaminergic-based strategies for PD management in general and for the L-DOPA-induced diskynesia (LID) in particular. Overall, we provide here an update of the current state of the art of these mGlu5 receptor-based approaches that are under clinical study as agents devoted to alleviate PD symptoms.

Determination of Ni(II) in metal alloys by spectrophotometry UV-Vis using dopasemiquinone

A spectrophotometric method was proposed for Ni(II) determination in alloys using a dopa-semiquinone (L-1) to form [Ni(II)(L1-)3]1-, ε = 9.3 x 10³ L mol-1 cm-1. The optimal conditions for the determination were: wavelength 590 nm, temperature 25 °C, reaction time 45 min and pH 7.5. The Beer's law was obeyed for nickel from 3.33 x 10-5 to 1.78 x 10-4 mol L-1. The method was applied to complex samples, such as inox, nickel-titanium and cobalt-chromium alloys. A study of the potential interferents revealed that Mn was the major interferent. The limit of detection and quantification were 2.88 x 10-5 mol L-1 and 3.06 x 10-5 mol L-1, respectively.

Spectrophotometric determination of catecholamine using vanadium and eriochrome cyanine r

A highly sensitive spectrophotometric method for the analysis of catecholamine drugs; L-dopa and methyldopa, is described. The analysis is based on the reaction of drug molecules with vanadium (V) which is reduced to vanadium (IV) and form complex with eriochrome cyanine R to give products having maximum absorbance (lmax) at 565 nm. Beer's law is obeyed in the range 0.028-0.84 and 0.099-0.996 mg mL-1 for L-dopa and methyldopa, respectively. The statistical analysis as well as comparison with reported methods demonstrated high precision and accuracy of the proposed method. The method was successfully applied in the analysis of pharmaceutical preparations.

Fármacos multifuncionais: monoamina oxidase e α-sinucleína como alvos terapêuticos na doença de Parkinson

Parkinson's disease (PD) is a neurodegenerative disorder associated to selective degeneration of dopaminergic neurons caused by an intricate relationship among dopamine metabolism, oxidative stress and α-synuclein fibrillation. Most therapies for PD have focused on dopamine replacement through the use of both monoamine oxidase inhibitors (MAOIs) and dopamine precursor L-dopa. Interestingly, certain MAOIs have a broad spectrum of action including anti-fibrillogenic properties in α-synuclein aggregation. Herein we revisit the chemical properties of MAOIs and their action on important targets associated with PD, notably α-synuclein fibrillation and dopamine metabolism, discussing the strategies associated with the development of multi-target drugs for neurodegenerative diseases.

Ultrastructural calli analysis of Inga vera Willd. subsp. Affinis (DC.) T.D. Penn

Subcellular changes are relevant to understand plant organogenesis and embryogenesis in the early stages of cell development. The cytology during cell development in tissue culture is however still poorly characterized. This study aimed to characterize the ultrastructural differences related to callogenesis of anthers, ovaries, leaf and nodal segments of Inga vera Willd. subsp. Affinis (DC.) T.D. Penn. Flower buds, nodal segments and leaves were disinfected and inoculated in test tubes containing MS medium with 3% sucrose and 4.5µM 2.4-D, except for leaf callogenesis, where 9µM of this auxin was used, and for the callogenesis of anthers and ovaries, where the culture medium was enriched with 0.25% activated charcoal and 90µM PVP. After 45 days in culture medium, the anther, ovary, leaf and nodal segment calli were fixed in Karnovisky and prepared for visualization by scanning and transmission electron microscopy. Ultrastructural differences were observed among the callus cells of anthers, ovaries, segments and leaves. There was no evidence of somatic embryo formation in the anther, leaf and nodal segment calli, in spite of some embryogenic characteristics in the cells. The ovary calli, with indications of embryo formation, seem to be the most responsive explant source for embryogenesis.

Lapsen sosiaalisen kompetenssin tukeminen – vanhempien epäsuorat ja suorat vaikutustavat

Tässä tutkimuksessa tarkastellaan vanhempien havaintoja ja käsityksiä lapsen sosiaalisesta kompetenssista. Lapsen sosiaalisesta kompetenssista tarkastelun kohteena ovat erityisesti vertaissuhteet, sosiaaliset taidot ja sosiaalinen käyttäytyminen. Tarkoituksena on selvittää vanhempien näkemyksiä lapsen sosiaalisesta verkostosta ja lapsesta sosiaalisena toimijana. Kiinnostuksen kohteena on myös, miten vanhemmat vaikuttamaan lapsen sosiaaliseen kompetenssiin. Vanhempien vaikutuksessa voidaan erottaa epäsuora ja suora vaikutus. Vanhempien epäsuoraan vaikutukseen kuuluvat perheen sosioekonomiset tekijät, vanhemmuuteen ja lastenkasvatukseen liittyvät käytännöt sekä lapsen ja vanhemman välinen vuorovaikutus. Suora vaikutus sisältää vanhempien eri roolit ja tehtävät sosiaalisen kompetenssin edistämiseksi. Vanhempien epäsuorilla ja suorilla vaikutustavoilla on havaittu olevan merkittävää vaikutusta lapsen sosiaalisen kompetenssiin muotoutumiseen ja sen laatuun. Tutkimuksessa selvitetään vertaissuhdeongelmaisten ja ei-ongelmaisten lasten vanhempien välisiä eroja näissä vaikutustavoissa. Tutkimuksessa hyödynnetään kyselylomake- ja haastatteluaineistoja. Kyselylomakeaineisto (N=156) kerättiin ”Origins of Exclusion in Early Childhood”-tutkimusprojektissa, jossa tutkittiin lasten vertaissuhteita, sosiaalisia taitoja sekä sosiaalista käyttäytymistä kolmen vuoden seurantatutkimuksena päiväkodista kouluun. Perhekysely toteutettiin lasten ollessa kuusivuotiaita. Vanhempien haastatteluaineisto (N=55) koostuu projektissa mukana olleiden lasten vanhempien teemahaastatteluista. Perhekyselyä analysoidaan tilastollisin analyysimenetelmin. Laadullisen aineiston analyysimenetelmänä käytetään sisällönanalyysia. Vanhempien käsityksissä lasten sosiaalinen verkosto rakentui kotiympäristössä, koulussa, päiväkodissa sekä suvun ja harrastusten parissa muodostuneista suhteista. Tutkimustulosten perusteella on havaittavissa, että vertaissuhdeongelmaisten ja ei-ongelmaisten lasten sosiaaliset verkostot ovat osin erilaiset. Vanhempien arviointien mukaan myös lasten sosiaalisissa taidoissa, käyttäytymisessä, asennoitumisessa sosiaaliseen kanssakäymiseen sekä ryhmään ja leikkeihin liittyvissä strategioissa on eroavaisuuksia. Tutkimuksen pohjalta voidaan todeta, että vanhemmat pystyvät arvioimaan hyvin yksityiskohtaisesti lapsensa sosiaalisia taitoja ja käyttäytymistä. Kaikilla vanhemmilla ei kuitenkaan ollut riittävästi tietoa lasten kaveripiiristä tai sen laadusta eikä lasten sosiaalisesta orientaatiosta. Vanhempien epäsuorissa vaikutustavoissa oli eroja, mutta myös yhtäläisyyksiä. Sosioekonomisia tekijöitä koskevan tarkastelun perusteella vertaissuhdeongelmaisten lasten perheiden taloudelliset ongelmat, isien työttömyys ja lapsen erityisen tuen tarve olivat yhteydessä lapsen sosiaalisten suhteiden ongelmiin. Lähes kaikki tutkimukseen osallistuneet vanhemmat kokivat vanhemmuuden ja kasvatustehtävän kuitenkin hyvin myönteisenä ja tyytyväisyyttä tuottavana asiana elämässä. Valtaosa vanhemmista piti lapsen ja aikuisen välistä vuorovaikutusta positiivisena, vaikka lapsen kanssa ei aina ollutkaan helppoa tulla toimeen. Tyytyväisyydestä huolimatta äidit näkivät itsessään enemmän kehittymisen tarpeita vanhempana kuin isät. Vanhemmuudessa korostuivat ohjaaminen ja kontrolli, mutta myös hoiva, lämpö ja vastavuoroisuus. Hoiva ja lämpö sekä rajojen asettaminen askarruttivat vanhempia suuresti. Vertaissuhdeongelmaisten lasten vanhemmat tarvitsisivat opastusta ohjaavan vanhemmuuden löytämiseksi. Vertaissuhdeongelmaisten lasten vanhemmat kuvasivat kasvatuksen kuormittavuutta, ajan puutetta sekä muuntuvaa isyyttä ja äitiyttä ei-ongelmaisten lasten vanhempia enemmän. Työn ja perheen yhteensovittamisen vaikeus tuli myös esille vanhempien kuvauksissa. Kyvykkään vanhemmuuden kannalta epävirallinen läheisistä muodostunut tukiverkosto on tärkeä vanhemman apu ja kasvatuksen turva. Ensisijaisena tukitahona on epävirallinen verkosto, joka koostuu ystävistä, tuttavista, työtovereista, puolisosta ja omista vanhemmista. Vertaissuhdeongelmaisten lasten vanhempien mukaan arjen tukea ei kuitenkaan ole aina saatavilla, eikä tukiverkosto tyydyttänyt vanhempia. Vanhempien käsityksissä perheen vuorovaikutus sujui hyvin ja vastuu kodista ja kasvatustehtävästä oli molemmilla vanhemmilla tasavertaisesti. Käytännön vastuu kasvatuksesta sekä erilaisten taitojen opettamisesta lapsille kuului äitien tehtäviin. Vanhempien näkemyksissä lapsen sosiaalinen maailma rakentui lähiympäristön tarjoamista mahdollisuuksista. Vanhempien suoriin vaikutustapoihin liittyvien tulosten mukaan vanhemmat pitävät harrastuksia merkittävänä sosiaalista kompetenssia edistävänä tekijänä. Ei-ongelmaisilla lapsilla oli enemmän ja monipuolisempia harrastuksia kuin ongelmaisilla lapsilla. Vaikka vanhemmat eivät mieltäneetkään omaa toimintaansa kaveripiiriin ohjaamiseksi, heillä useinkin oli runsaasti erilaisia rooleja ja tehtäviä sosiaalisten suhteiden ja sosiaalisten taitojen opettamisessa. Lapsen sosiaalisten suhteiden organisoinnissa, ohjaamisessa, valvonnassa ja neuvonnassa oli vanhempiryhmien välillä eroja. Vertaissuhdeongelmaisten lasten vanhemmat eivät olleet riittävän hyvin perillä lapsen taidoista ja kyvykkyydestä tuottaakseen oikea-aikaista ohjausta ja tukeakseen lasta riittävästi sopivilla tavoilla. Vanhempien toimintaa näyttää ohjaavan vakaasti se uskomus, että lapsen kaverisuhteet ovat pelkästään hänen oma asiansa. Vanhemmat arvostivat paljon sosiaalisia taitoja ja pyrkivät opettamaan niitä lapsilleen systemaattisesti. Ohjaamisen tavoissa vanhemmat poikkesivat toisistaan. Tutkimus antaa viitteitä, että eiongelmaisten lasten vanhemmat ovat sensitiivisempiä, vastavuoroisempia ja lapsen näkökulmaa ja tarpeita lähtökohtanaan pitäviä sekä tunnetaitoihin ja tunnetilan käsittelyyn keskittyvämpiä kuin ongelmaisten lasten vanhemmat. Vanhempien ja lasten vuorovaikutuksen vaikeudet, ylimalkainen ja epäjohdonmukainen sosiaalisten taitojen, käyttäytymisen sekä suhteiden ohjaus sekä monet perheen voimavarojen puutteet voivat haitata vakavasti lapsen sosiaalisen kompetenssin kehittymistä. Vanhemmilla on kuitenkin taitoa arvioida lapsensa sosiaalista kyvykkyyttä ja halua toimia hyvin kasvatustehtävässään. Vanhemmuuden tukemisessa olisivatkin tärkeitä epävirallisen tukiverkoston lisäksi lähiympäristön perheille ja lapsille suunnatut palvelut, monitahoinen yhteistyö perheiden, lasten ja heidän kanssaan työskentelevien ammattilaisten kesken sekä yhteiskunnan perheitä koskevat säädökset ja tukitoimet.

18F-Labeled Presynaptic Dopaminergic Tracers. Synthesis, Radiochemical Analyses, and Preclinical Evaluation of [18F]FDOPA and [18F]CFT

Dysfunction of the dopaminergic system in brain is involved in several pathological conditions such as Parkinson’s disease and depression. 2β-Carbomethoxy-3β-(4-[18F] fluorophenyl)tropane ([18F]CFT) and 6-[18F]fluoro-L-dopa ([18F]FDOPA) are tracers for imaging the dopaminergic function with positron emission tomography (PET). Peripheral uptake of [18F]FDOPA is also used in the localization and diagnosis of neuroendocrine tumors. [18F]FDOPA and [18F]CFT can be synthesized by electrophilic fluorodestannylation. However, the specific radioactivity (SA) in the electrophilic fluorination is low with traditional synthetic methods. In this study, [18F]FDOPA and [18F]CFT were synthesized using post-target-produced [18F]F2 as an electrophilic fluorination agent. With this method, tracers are produced with sufficient SA for neuroreceptor studies. Specific aims in this study were to replace Freon-11 in the production of [18F]FDOPA due to the ozone depleting properties of this solvent, to determine pharmacological specificity and selectivity of [18F]CFT with respect to monoamine transporters, and to compare the ability of these tracers to reflect the degree of nigral neuronal loss in rats in which the dopaminergic system in the brain had been unilaterally destroyed by 6- OHDA. Post-target-produced [18F]F2 was successfully used in the production of [18F]FDOPA and [18F]CFT. The SA achieved was substantially higher than in previous synthetic methods. Deuterated compounds, CD2Cl2, CDCl3 and C3D6O, were found to be suitable solvents for replacing Freon-11. Both [18F]FDOPA and [18F]CFT demonstrated nigrostriatal dopaminergic hypofunction and correlated with the number of nigral dopaminergic neurons in the 6-OHDA lesioned rat. However, the dopamine transporter (DAT) tracer [18F]CFT was more sensitive than the dopamine synthesis tracer [18F]FDOPA in detecting these defects because of the higher non-specific uptake of [18F]FDOPA. [18F]CFT can also be used for imaging the norepinephrine transporter (NET) because of the specific uptake into the locus coeruleus. The observation that [18F]CFT exhibits specific uptake in the pancreas warrants further studies in humans with respect to potential utility in pancreatic imaging

Superoxide dismutase 3-mediated cell survival and proliferation

This dissertation studies the signaling events mediated by the extracellular superoxide dismutase (SOD3). SOD3 is an antioxidant enzyme which converts the harmful superoxide into hydrogen peroxide. Overproduction of these reactive oxygen species (ROS) in the cellular environment as a result of tissue injury or impaired antioxidant defense system has detrimental effects on tissue integrity and function. However, especially hydrogen peroxide is also an important signaling agent. Ischemic injury in muscle causes acute oxidative stress and inflammation. We investigated the ability of SOD3 to attenuate ischemia induced inflammation and to promote recovery of skeletal muscle tissue. We found that SOD3 can downregulate the expression of several inflammatory cytokines and cell adhesion molecules thus preventing the accumulation of oxidant-producing inflammatory cells. Secondly, SOD3 was able to promote long-term activation of the mitogenic Erk pathway, but increased only briefly the activity of pro-survival Akt pathway at an early stage of ischemic inflammation, thus reducing apoptosis. SOD3 is a prominent antioxidant in the thyroid gland where oxidative stress is constantly present. We investigated the role of SOD3 in normal thyroid follicular cells and the changes in its expression in various hyperproliferative disorders. We first showed that SOD3 is TSH-responsive which indicated its participation in thyroid function. Its principal function seems to be in follicular cell proliferation since knockdown cells were deficient in proliferation. Additionally, it was overexpressed in goiter tissue. However, SOD3 was consistently downregulated in thyroid cancer cell lines and tissues. In conclusion, SOD3 is involved in tissue maintenance, cell proliferation and inflammatory cell migration. Its mechanisms of action are the activation of known proliferation/survival pathways, inhibition of apoptosis and regulation of adhesion molecule expression.

Satama sydämettömässä maailmassa?

Kirjallisuusarvostelu

JNK, a versatile regulator of kinesin-1 transport and cytoskeleton dynamics

C-Jun N-terminal kinase (JNK) is traditionally recognized as a crucial factor in stress response and inducer of apoptosis upon various stimulations. Three isoforms build the JNK subfamily of MAPK; generally expressed JNK1 and JNK2 and brain specific JNK3. Degenerative potency placed JNK in the spotlight as potential pharmacological option for intervention. Unfortunately, adverse effects of potential drugs and observation that expression of only JNK2 and JNK3 are induced upon stress, restrained initial enthusiasm. Notably, JNK1 demonstrated atypical high constitutive activity in neurons that is not responsive to cellular stresses and indicated existence of physiological activity. This thesis aimed at revealing the physiological functions of JNK1 in actin homeostasis through novel effector MARCKS-Like 1 (MARCKSL1) protein, neuronal trafficking mediated by major kinesin-1 motor protein and microtubule (MT) dynamics via STMN2/SCG10. The screen for novel physiological JNK substrates revealed specific phosphorylation of C-terminal end of MARCKSL1 at S120, T148 and T183 both ex vivo and in vitro. By utilizing site-specific mutagenesis, various actin dynamics and migrations assays we were able to demonstrate that JNK1 phosphorylation specifically facilitates F-actin bundling and thus filament stabilisation. Consecutively, this molecular mechanism was proved to enhance formation of filopodia; cell surface projections that allow cell sensing surrounding environment and migrate efficiently. Our results visualize JNK dependent and MARCKSL1 executed induction of filopodia in neurons and fibroblast indicating general mechanism. Subsequently, inactivation of JNK action on MARCKSL1 shifts cellular actin machinery into lamellipodial dynamic arrangement. Tuning of actin cytoskeleton inevitably melds with cell migration. We observed that both active JNK and JNK pseudo-phosphorylated form of MARCKSL1 reduce actin turnover in intact cells leading to overall diminished cell motility. We demonstrate that tumour transformed cells from breast, prostate, lung and muscle-derived cancers upregulate MARCKSL1. We showed on the example of prostate cancer PC-3 cell line that JNK phosphorylation negatively controls MARCKSL1 ability to induce migration, which precedes cancer cell metastasis. The second round of identification of JNK physiological substrates resulted in detection of predominant motor protein kinesin-1 (Kif5). Mass spectrometry detailed analysis showed evident endogenous phosphorylation of kinesin-1 on S176 within motor domain that interacts with MT. In vitro phosphorylation of bacterially expressed kinesin heavy chain by JNK isoforms displayed higher specificity of JNK1 when compared to JNK3. Since, JNK1 is constitutively active in neurons it signified physiological aspect of kinesin-1 regulation. Subsequent biochemical examination revealed that kinesin-1, when not phosphorylated on JNK site, exhibits much higher affinity toward MTs. Expression of the JNK non-phosphorable kinesin-1 mutant in intact cells as well as in vitro single molecule imaging using total internal reflection fluorescence microscopy indicated that the mutant loses normal speed and is not able to move processively into proper cellular compartments. We identify novel kinesin-1 cargo protein STMN2/SCG10, which along with known kinesin-1 cargo BDNF is showing impaired trafficking when JNK activity is inhibited. Our data postulates that constitutive JNK activity in neurons is crucial for unperturbed physiologically relevant transport of kinesin-1 dependant cargo. Additionally, my work helps to validate another novel physiological JNK1 effector STMN2/SCG10 as determinant of axodendritic neurites dynamics in the developing brain through regulation of MT turnover. We show successively that this increased MT dynamics is crucial during developmental radial migration when brain layering occurs. Successively, we are able to show that introduction of JNK phosphorylation mimicking STMN2/SCG10 S62/73D mutant rescues completely JNK1 genetic deletion migration phenotype. We prove that STMN2/SCG10 is predominant JNK effector responsible for MT depolymerising activity and neurite length during brain development. Summarizing, this work describes identification of three novel JNK substrates MARCKSL1, kinesin-1 and STMN2/SCG10 and investigation of their roles in cytoskeleton dynamics and cargo transport. This data is of high importance to understand physiological meaning of JNK activity, which might have an adverse effect during pharmaceutical intervention aiming at blocking pathological JNK action.

How (not) to interview children : interviews with young children in sexual abuse investigations in Finland

The present thesis had two main objectives: The first was to assess how child sexual abuse (CSA) interviews in Finland are conducted through analysing the interviewing techniques applied and the language used by the interviewers, as well as to suggest ways to improve interviews if they were found to have deficiencies. The second main aim was to contribute to the growing research corpus concerning CSA interviews, in particular, by addressing how interviewers follow up information provided by the child, by analysing whether child health care professionals would use childadapted language, and by studying the kind of modifications in the verbal behaviour of interviewers and children that were associated with a) repeated interviews, b) a support person’s presence at the interview, and c) the use of anatomically detailed dolls. Two complementary samples of CSA interviews were analysed. The first one was composed of child interviews with 3-12-year-old children (N = 27) that had been considered problematic by lawyers or other involved professionals (Studies I and IV). The second sample consisted of unselected interviews (N = 43) with children aged 3 to 8 years conducted in a number of hospitals in different parts of the country (Studies II and III). Study I: The verbal interaction between interviewer and child was analysed in a sample of interviews that had been considered to be problematic by involved professionals. Results showed that interviewers used inappropriate questioning techniques, relying on option-posing, specific suggestive and unspecific suggestive questions to a significant extent, these comprising around 50% of all interviewer utterances. The proportion of invitations, which the research community recommends interviewers to rely on, was strikingly low. Invitations and directive utterances were associated with an increase in informative responses by the child in terms of response type, number of new details reported, as well as length of response. The opposite was true for option-posing and suggestive utterances. Longer questions by the interviewer (in number of words) often rendered no reply from the child, whereas shorter questions were followed by descriptive answers. Even after the child had provided an informative answer, interviewers failed to follow up the information in an adequate way and instead continued to rely on focused and leading questions. Study II: Due to the possible bias of the sample analysed in Study I, the most important analyses were rerun with the unselected sample and reported separately. Results were quite similar between the two studies, indicating that the problems observed in Study I, with interviewers relying on option-posing and suggestive questions to a significant extent, are likely to be general and not specific for those interviews. Even if suggestive questions were slightly less and invitations slightly more common in this sample than in the previous study, almost half of the interviewer questions were still optionposing or suggestive, and also in this sample, interviewers failed to follow up information by the child in a facilitating manner. Differentiating between judicial and contextual details showed that while facilitators, invitations, and directive utterances elicited more contextual than judicial details, the opposite was true for specific suggestive utterances. These results might be explained by the reluctance of children to describe sexual details related to the abuse events. Alternatively, they may also be due to children describing incorrect sexual details as a result of suggestive interviewing techniques. Study III: This study examined features of the language used by the interviewers. Interviewer utterances included multiple questions, long statements, complicated grammar and concepts, as well as unclear references to persons and situations. More than a fifth of the interviewer utterances were coded as belonging to at least one of these categories. The results suggest that even professionals who are experienced in interacting with children may have difficulties in using a child-sensitive language, adding to the pool of studies showing similar problems to occur in legal hearings with children conducted by lawyers. As children rarely comment on, or even recognise, their lack of comprehension, the use of a language that is too complex can have detrimental consequences for the outcomes of investigative interviews. Interviewers used different approaches to introduce the topic of abuse. While 15% of the children spontaneously addressed the topic of abuse, probably indicating that they felt confident with the interviewer and the situation, in almost 50% of the cases, the interviewer introduced the topic of abuse in a way that can be considered leading. Interviews were characterised by a lack of structure, apparent in frequent rapid switches of topic by the interviewer. This manner was associated with a decrease in the number of new details provided by the children. Study IV: This study analysed possible changes in the interview dynamics associated with repeated interviewing, the presence of a support person (related to the child), and the use of anatomically detailed (AD) dolls. Repeated interviewing, in combination with suggestive questions, has previously been found to seriously contaminate children’s accounts. In the present material, interviewers used significantly more suggestive utterances in the repeated condition, thus endangering the reliability of the children’s reports. Few studies have investigated the effects of a support person’s presence at the interview. The results of the present study showed that interviewers talked more and children provided less information when a support person was present. Supporting some earlier findings regarding the use of AD dolls, the present results showed that using AD dolls was associated with longer interviewer utterances and shorter, less responsive, and less detailed child responses. Interviewers used up to five times more unspecific suggestive utterances when dolls were used, for instance through repeatedly asking the child to show “what really happened” with the dolls. Conclusion: The results indicate that CSA interviews in Finland are not conducted in a manner that follows best practice as defined by the research community and as stated in a number of guidelines. When comparing these questioning strategies with the recommendations, which have been predominant in the field for more than ten years now, it can be concluded that the interviews analysed were conducted in a manner that undermines the possibility to elicit an uncontaminated and accurate narrative from the children. A particularly worrying finding was the fact that interviewers did not follow up relevant information by the children in an adequate way. A number of clinical implications can be drawn from the results, particularly concerning the need for improvement in the quality of CSA interviews. There is convincing research regarding how to improve CSA interviews, notably through training forensic child interviewers to use a structured interviewing protocol, and providing them with continuous supervision and feedback. Allocating appropriate resources to improve the quality of forensic child interviews is a matter of protecting the rights of all persons involved in CSA investigations, in particular those of the children.

Enhancing the development of military capabilities by a systems approach

The political environment of security and defence has changed radically in the Western industrialised world since the Cold War. As a response to these changes, since the beginning of the twenty-first century, most Western countries have adopted a ‘capabilities-based approach’ to developing and operating their armed forces. More responsive and versatile military capabilities must be developed to meet the contemporary challenges. The systems approach is seen as a beneficial means of overcoming traps in resolving complex real -world issues by conventional thinking. The main objectives of this dissertation are to explore and assess the means to enhance the development of military capabilities both in concept development and experimentation (CD&E) and in national defence materiel collaboration issues. This research provides a unique perspective, a systems approach, to the development areas of concern in resolving complex real-world issues. This dissertation seeks to increase the understanding of the military capability concept both as a whole and with in its life cycle. The dissertation follows the generic functionalist systems methodology by Jackson. The methodology applies a comprehensive set of constitutive rules to examine the research objectives. This dissertation makes contribution to current studies about military capability. It presents two interdepen dent conceptual capability models: the comprehensive capability meta-model (CCMM) and the holistic capability life cycle model (HCLCM). These models holistically and systematically complement the existing, but still evolving, understanding of military capability and its life cycle. In addition, this dissertation contributes to the scientific discussion of defence procurement in its broad meaning by introducing the holistic model about the national defence materiel collaboration between the defence forces, defence industry and academia. The model connects the key collaborative mechanisms, which currently work in isolation from each other, and take into consideration the unique needs of each partner. This dissertation contributes empirical evidence regarding the benefits of enterprise architectures (EA) to CD&E. The EA approach may add value to traditional concept development by increasing the clarity, consistency and completeness of the concept. The most important use considered for EA in CD&E is that it enables further utilisation of the concept created in the case project.