Construção de uma câmara para monitoramento in situ do processo de secagem de geis e sólidos porosos

The present work deals with the design and construction of an equipment for studying the drying process of gels and solids. The equipment was built using mainly a commercial oven and others peripherals for programmed control of temperature, humidity and data acquisition. The system allows the in situ monitoring of the gel drying, registering the weight loss, and the shrinkage of sample and the oven temperatures. This set of allowed parameters makes possible to establish the main steps involved in the drying of meso and nanostructured porous materials.

Hydrogels of cellulose acetate crosslinked with pyromellitic dianhydride: part I: synthesis and swelling kinetics

This work describes the synthesis of hydrogels of cellulose acetate (AC) crosslinked with 1,2,4,5-benzenotetracarboxylic dianhydride (PMDA). The crosslinking reaction was monitored by FTIR. Analysis of aromatic fragments from the alkaline hydrolysis of the gels by UV spectroscopy indicated that an increase in the stoichiometric ratio of dianhydride resulted in higher degrees of crosslinking. The non-porous nature of the gels was confirmed by analysis of nitrogen adsorption. Water absorption isotherms showed that as the temperature and degree of crosslinking increased, the percentage of water absorbed at equilibrium (%Seq) also increased. The hydrogels presented second order swelling kinetics.

Cristalización de zeolita beta mediante conversión asistida por vapor: influencia de los parámetros de síntesis

In the present work, beta zeolites were prepared by an alternative route called steam-assisted conversion (SAC). Several zeolites were synthesized using amorphous dry gels with a low SDA concentration (0.09 mol, TEAOH). Temperature and crystallization time were the main parameters studied. X-ray diffraction (XRD), infrared spectroscopy (IR) and scanning electron microscopy (SEM) were the characterization techniques employed. The zeolites prepared showed mixed phases such as beta, MTW and ZSM-5 while only one sample treated at 150ºC with 24 h of crystallization time showed a pure ZSM-5 phase (SAC-5). These preliminary results serve as a starting point for optimizing the synthesis of a specific type of zeolite using the SAC method.

A NOVEL POLYACRYLAMIDE-BASED HYDROGEL CROSSLINKED WITH CELLULOSE ACETATE AND PREPARED BY PRECIPITATION POLYMERIZATION

The synthesis of polyacrylamide-cellulose acetate hydrogels by precipitation polymerization in acetone solution is reported herein. These hydrogels exhibit smaller swelling ratios and larger compression moduli than homo polyacrylamide hydrogels. For cellulose acetate concentrations above 20 wt.%, hydrogels with N,N'-methylenebisacrylamide as a crosslinker exhibit swelling ratios and compression moduli similar to those of the hydrogels without the crosslinker. A possible explanation for this behavior is that cellulose acetate crosslinks polyacrylamide via free-radical reaction. The hydrogels obtained without the N,N'-methylenebisacrylamide crosslinker exhibit compression moduli up to 1.7 MPa, making them suitable for tissue engineering applications such as cartilage replacement.

INFLUÊNCIA DO CATALISADOR EM SOLUÇÕES POLIMÉRICAS PARA O PREPARO DE GÉIS ORGÂNICOS DE RESORCINOL-FORMALDEÍDO

Resorcinol-formaldehyde (RF) organic gels have been extensively used to produce carbon aerogels. The organic gel synthesis parameters greatly affect the structure of the resulting aerogel. In this study, the influence of the catalyst quantity on the polymeric solution sol-gel process was investigated. Sodium carbonate was used as a basic catalyst. RF gels were synthesized with a resorcinol to formaldehyde molar ratio of 0.5, a resorcinol to catalyst (R/C) molar ratio equal to 50 or 300, and a resorcinol to solvent ratio of 0.1 g mL-1. The sol-gel process was evaluated in situ by Fourier transform infrared spectroscopy using a universal attenuated total reflectance sensor and measurements of the kinematic viscosity. The techniques showed the evolution of the sol-gel process, and the results showed that the lower catalyst quantity induced a higher gel point, with a lower viscosity at the gel point. Differential scanning calorimetry was used to investigate the thermal behavior of the RF dried gel, and results showed that the exothermic event related to the curing process was shifted to higher temperatures for solutions containing higher R/C ratios.

Histopathological features of infections caused by Fusarium oxysporum and F. solani in purple passionfruit plants (Passiflora edulis Sims)

The purple passionfruit plant, Passifloraedulis Sims, ranks second in fruit exportation in Colombia, and its main destination is the European market. However, its production is affected by several diseases, including fusariosis. This paper presents the histopathological features of different tissues affected by the pathogens Fusarium oxysporum and Fusarium solani. Both microorganisms produce similar responses on the plant: colonization of xylem vessels by hyphae and microconidia, hypertrophy and hyperplasia of the cambium, xylem and phloem; destruction of xylem fibers and amyloplasts in parenchymatous cells; and production of gels by the plant. However, there are differences in the colonization mechanism, F. solani penetrates and is concentrated especially at the collar zone, while F. oxysporum penetrates the roots and moves through the vascular system to colonize the plant.

EDTA- and DTPA-functionalized silica gel and chitosan adsorbents for the removal of heavy metals from aqueous solutions

Adsorbents functionalized with chelating agents are effective in removal of heavy metals from aqueous solutions. Important properties of such adsorbents are high binding affinity as well as regenerability. In this study, aminopolycarboxylic acid, EDTA and DTPA, were immobilized on the surface of silica gel, chitosan, and their hybrid materials to achieve chelating adsorbents for heavy metals such as Co(II), Ni(II), Cd(II), and Pb(II). New knowledge about the adsorption properties of EDTA- and DTPA-functionalizedadsorbents was obtained. Experimental work showed the effectiveness, regenerability, and stability of the studied adsorbents. Both advantages and disadvantages of the adsorbents were evaluated. For example, the EDTA-functionalized chitosan-silica hybrid materials combined the benefits of the silica gel and chitosan while at the same time diminishing their observed drawbacks. Modeling of adsorption kinetics and isotherms is an important step in design process. Therefore, several kinetic and isotherm models were introduced and applied in this work. Important aspects such as effect of error function, data range, initial guess values, and linearization were discussed and investigated. The selection of the most suitable model was conducted by comparing the experimental and simulated data as well as evaluating the correspondence between the theory behind the model and properties of the adsorbent. In addition, modeling of two-component data was conducted using various extended isotherms. Modeling results for both one- and twocomponent systems supported each other. Finally, application testing of EDTA- and DTPA-functionalized adsorbents was conducted. The most important result was the applicability of DTPA-functionalized silica gel and chitosan in the capturing of Co(II) from its aqueous EDTA-chelate. Moreover, these adsorbents were efficient in various solution matrices. In addition, separation of Ni(II) from Co(II) and Ni(II) and Pb(II) from Co(II) and Cd(II) was observed in two- and multimetal systems. Lastly, prior to their analysis, EDTA- and DTPA-functionalized silica gels were successfully used to preconcentrate metal ions from both pure and salty waters

Immobilization of Burkholderia cepacia Lipase: Kinetic Resolution in Organic Solvents, Ionic Liquids and in Their Mixtures

Immobilization of Burkholderia cepacia Lipase: Kinetic Resolution in Organic Solvents, Ionic Liquids and in Their Mixtures Biocatalysis opens the door to green and sustainable processes in synthetic chemistry allowing the preparation of single enantiomers, since the enzymes are chiral and accordingly able to catalyze chemical reactions under mild conditions. Immobilization of enzymes enhances process robustness, often stabilizes and activates the enzyme, and enables reuse of the same enzyme preparation in multiple cycles. Although hundreds of variations of immobilization methods exist, there is no universal method to yield the highly active, selective and stable enzyme catalysts. Therefore, new methods need to be developed to obtain suitable catalysts for different substrates and reaction environments. Lipases are the most widely used enzymes in synthetic organic chemistry. The literature part together with the experimental part of this thesis discusses of the effects of immobilization methods mostly used to enhance lipase activity, stability and enantioselectivity. Moreover, the use of lipases in the kinetic resolution of secondary alcohols in organic solvents and in ionic liquids is discussed. The experimental work consists of the studies of immobilization of Burkholderia cepacia lipase (lipase PS) using three different methods: encapsulation in sol-gels, cross-linked enzyme aggregates (CLEAs) and supported ionic liquids enzyme catalysts (SILEs). In addition, adsorption of lipase PS on celite was studied to compare the results obtained with sol-gels, CLEAs and SILEs. The effects of immobilization on enzyme activity, enantioselectivity and hydrolysis side reactions were studied in kinetic resolution of three secondary alcohols in organic solvents, in ionic liquids (ILs), and in their mixtures. Lipase PS sol-gels were shown to be active and stable catalysts in organic solvents and solvent:IL mixtures. CLEAs and SILEs were highly active and enantioselective in organic solvents. Sol-gels and SILEs were reusable in several cycles. Hydrolysis side reaction was suppressed in the presence of sol-gels and CLEAs.

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.

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.

First identification of natural infection of Rickettsia rickettsii in the Rhipicephalus sanguineus tick, in the State of Rio de Janeiro

The Brazilian Spotted Fever (BSF) is a zoonotic disease caused by Rickettsia rickettsii and transmitted by ticks of the genus Amblyomma, more frequently, Amblyomma cajennense. The aim of this paper was to report the first molecular detection of R. rickettsii on R. sanguineus naturally infected in Rio de Janeiro, Brazil. Ticks were collected from dogs in a rural region of Resende municipality, Rio de Janeiro State, Brazil (22º30'9.46"S, 44º42'44.29"WO), where occurred five human cases of BSF in 2006. The ticks were identified under a stereoscopic microscope and separated in pools by stages, species and sex. DNA extraction was carried out using QIAamp DNA Mini Kit (QIAGEN®). The DNA was submitted to PCR amplification using 04 set of primers: Rr190.70p/Rr190.602n (OmpA, 532bp), BG1-21/BG2-20 (OmpB, 650bp), Tz15/Tz16 (17 kDa protein-encoding gene, 246bp) and RpCS.877p/RpCS.1258n (gltA, 381bp). PCR products were separated by electrophoresis on 1% agarose gels and visualized under ultraviolet light with ethidium bromide. PCR products of the expected sizes were purified by QIAquick® and sequenced by ABI PRISM®. The generated nucleotide sequences were edited with using Bioedit® software and compared with the corresponding homologous sequences available through GenBank, using Discontiguous Mega Blast (http://www.ncbi.nlm.nih.gov). It was confirmed R. rickettsii by sequencing of the material (GenBank FJ356230). The molecular characterization of R. rickettsii in the tick R. sanguineus emphasizes the role of dogs as carriers of ticks from the environment to home. Moreover, this result suggests that there is a considerable chance for active participation of R. sanguineus as one of tick species in the transmission of R. ricketsii to human being in the Brazilian territory.

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.