Double grade – rakenneputkien puristuskestävyys

Double grade S420MH/S355J2H – rakenneputki on Ruukin kylmämuovattujen rakenneputkien vakioteräslaji. Se voidaan mitoittaa joko lujuusluokan S355 tai S420 mukaisesti. Teräslajin S355 mukaisesti mitoitettaessa on suunnittelu yksinkertaista. Painonsäästöä ja pidennettyjä jännevälejä haluttaessa käytetään lujuusluokan S420 mukaista mitoitusta. Työn tavoitteena oli selvittää kylmämuovattujen teräsrakenneputkien todellinen puristuskestävyys. Eurocode 3:n mukaan kylmämuovatut teräsrakenneputket kuuluvat nurjahduskäyrälle c. Tutkimukseen valittiin viisi eri profiilia olevaa rakenneputkea, joiden poikkileikkausluokat olivat 1, 2, 3 ja 4. Käytettäessä rakenneputkia puristussauvoina, on teräksen käyttö tehokkainta poikkileikkausluokassa 3, lähellä poikkileikkausluokkaa 4. Rakenneputkista laskettiin muunnetun hoikkuuden arvoilla 0.1, 0.5, 1.0 ja 1.5 koesauvojen pituudet kaikille profiileille. Valmistettiin kolme samanlaista koesauvaa jokaisesta koosta ja puristuskokeita suoritettiin yhteensä 57 kappaletta. Koesauvojen todelliset pituudet, alkukäyryydet ja poikkileikkaukset mitattiin. Ainestodistuksista saatiin materiaalin todelliset lujuudet. Laskettiin Eurocode 3:n mukaisesti kestävyydet nurjahduskäyrille a, b ja c. Laskennallisia kestävyyksiä verrattiin puristuskokeiden tuloksiin. Puristuskokeiden tulosten perusteella voidaan b-käyrää pitää oikeana profiileille 100x100x3, 150,150x5 ja 200x200x6. Profiili 150x150x5 kuuluu poikkileikkausluokkaan 2. Profiilit 100x100x3 ja 200x200x6 kuuluvat poikkileikkausluokkaan 4. Profiili 50x50x2 kuuluu nurjahduskäyrälle c. Profiilin poikkileikkausluokka on 1 ja aiemmat tutkimukset tukevat nurjahduskäyrän c käyttöä. Profiilista 300x300x8.8 ei saatu testattua täyttä sarjaa sen suuren kapasiteetin rikottua testilaitteiston, mutta puristuskokeiden perusteella se kuuluu nurjahduskäyrälle b. Profiili kuuluu poikkileikkausluokkaan 4.

Advanced analysis and design methods for preparative chromatographic separation processes

Preparative liquid chromatography is one of the most selective separation techniques in the fine chemical, pharmaceutical, and food industries. Several process concepts have been developed and applied for improving the performance of classical batch chromatography. The most powerful approaches include various single-column recycling schemes, counter-current and cross-current multi-column setups, and hybrid processes where chromatography is coupled with other unit operations such as crystallization, chemical reactor, and/or solvent removal unit. To fully utilize the potential of stand-alone and integrated chromatographic processes, efficient methods for selecting the best process alternative as well as optimal operating conditions are needed. In this thesis, a unified method is developed for analysis and design of the following singlecolumn fixed bed processes and corresponding cross-current schemes: (1) batch chromatography, (2) batch chromatography with an integrated solvent removal unit, (3) mixed-recycle steady state recycling chromatography (SSR), and (4) mixed-recycle steady state recycling chromatography with solvent removal from fresh feed, recycle fraction, or column feed (SSR–SR). The method is based on the equilibrium theory of chromatography with an assumption of negligible mass transfer resistance and axial dispersion. The design criteria are given in general, dimensionless form that is formally analogous to that applied widely in the so called triangle theory of counter-current multi-column chromatography. Analytical design equations are derived for binary systems that follow competitive Langmuir adsorption isotherm model. For this purpose, the existing analytic solution of the ideal model of chromatography for binary Langmuir mixtures is completed by deriving missing explicit equations for the height and location of the pure first component shock in the case of a small feed pulse. It is thus shown that the entire chromatographic cycle at the column outlet can be expressed in closed-form. The developed design method allows predicting the feasible range of operating parameters that lead to desired product purities. It can be applied for the calculation of first estimates of optimal operating conditions, the analysis of process robustness, and the early-stage evaluation of different process alternatives. The design method is utilized to analyse the possibility to enhance the performance of conventional SSR chromatography by integrating it with a solvent removal unit. It is shown that the amount of fresh feed processed during a chromatographic cycle and thus the productivity of SSR process can be improved by removing solvent. The maximum solvent removal capacity depends on the location of the solvent removal unit and the physical solvent removal constraints, such as solubility, viscosity, and/or osmotic pressure limits. Usually, the most flexible option is to remove solvent from the column feed. Applicability of the equilibrium design for real, non-ideal separation problems is evaluated by means of numerical simulations. Due to assumption of infinite column efficiency, the developed design method is most applicable for high performance systems where thermodynamic effects are predominant, while significant deviations are observed under highly non-ideal conditions. The findings based on the equilibrium theory are applied to develop a shortcut approach for the design of chromatographic separation processes under strongly non-ideal conditions with significant dispersive effects. The method is based on a simple procedure applied to a single conventional chromatogram. Applicability of the approach for the design of batch and counter-current simulated moving bed processes is evaluated with case studies. It is shown that the shortcut approach works the better the higher the column efficiency and the lower the purity constraints are.

Influence of the neural tube/notochord complex on MyoD expression and cellular proliferation in chicken embryos

Important advances have been made in understanding the genetic processes that control skeletal muscle formation. Studies conducted on quails detected a delay in the myogenic program of animals selected for high growth rates. These studies have led to the hypothesis that a delay in myogenesis would allow somitic cells to proliferate longer and consequently increase the number of embryonic myoblasts. To test this hypothesis, recently segmented somites and part of the unsegmented paraxial mesoderm were separated from the neural tube/notochord complex in HH12 chicken embryos. In situ hybridization and competitive RT-PCR revealed that MyoD transcripts, which are responsible for myoblast determination, were absent in somites separated from neural tube/notochord (1.06 and 0.06 10-3 attomol MyoD/1 attomol ß-actin for control and separated somites, respectively; P<0.01). However, reapproximation of these structures allowed MyoD to be expressed in somites. Cellular proliferation was analyzed by immunohistochemical detection of incorporated BrdU, a thymidine analogue. A smaller but not significant (P = 0.27) number of proliferating cells was observed in somites that had been separated from neural tube/notochord (27 and 18 for control and separated somites, respectively). These results confirm the influence of the axial structures on MyoD activation but do not support the hypothesis that in the absence of MyoD transcripts the cellular proliferation would be maintained for a longer period of time.

Esikiristetyn pulttiliitoksen vaikutus hitsausmuodonmuutoksiin

Tässä työssä tutkitaan propulsioyksikön kiinnitysrenkaan pulttiliitosten vaikutusta asen-nushitsauksesta aiheutuviin hitsausmuodonmuutoksiin. Hitsausmuodonmuutoksissa tutki-taan tärkeimpinä kohtina asennuslohkossa laakerin rajapintaa sekä kääntömoottorin kiinni-tyspintaa. Tutkimuksessa asennuslohkon hitsaaminen ja muodonmuutosten arvioiminen toteutettiin käyttämällä epälineaarista elementtimenetelmää. Ensisijaisena tavoitteena työssä on tutkia esikiristettyjen pulttiliitoksien vaikutusta raken-teen muodonmuutoksiin ja pohtia aiheutuvien siirtymien perusteella pulttien tarpeellisuutta rakenteessa. Tämän lisäksi vertaillaan pultillisen ja pultittoman kiinnitystavan eroavaisuuk-sia tuloksia analysoimalla. Saatujen tuloksien perusteella radiaaliset ja aksiaaliset siirtymät eivät olleet riittävän suuria aiheuttamaan haittoja rakenteen toimivuudelle kummassakaan mallissa. Lisäanalyysejä tar-kemmalla lämmöntuonnilla voidaan pitää tarvittavana pulttiliitoksien tarpeellisuuden tar-kemman testaamisen vuoksi.

Kylmälaitekoneikon runkorakenteen kehittäminen

Kylmälaitekoneikot ovat kylmäkomponentteja sisältäviä rakenteita, joiden avulla toteutetaan suurten tilojen, kuten elintarvikemyymälöiden sisäilman jäähdytys. Lisäksi koneikkojen avulla jäähdytetään matalampiin lämpötiloihin pienempiä kylmähuoneita. Osa koneikoista ottaa talteen kylmäprosessissa syntyvän lämmön, jota hyödynnetään tilojen lämmityksessä. Tämän diplomityön tavoitteena oli suunnitella ja mitoittaa kahdeksalle eri kylmälaitekoneikolle entistä kustannustehokkaammat runkorakenteet, jotka ovat niin kestäviä, että koneikkoja on mahdollista pinota tilan säästämiseksi kolme päällekkäin. Lisäksi runkorakenteilta vaadittiin helppoa kuljetettavuutta, hyviä kiinnitysominaisuuksia ja korroosionkestävyyttä. Aluksi työssä selvitettiin runkorakenteisiin kohdistuvat vaatimukset, jonka jälkeen materiaalin valinta tehtiin materiaaliin kohdistuvien vaatimusten perusteella. Rakenteiden palkit mitoitettiin tarvittavan taivutusvastuksen ja kiepahduksen mukaan. Pilarit puolestaan mitoitettiin nurjahduksen ja kaksiaksiaalisen taivutustilan perusteella. Tämän jälkeen mitoitettiin eri sauvojen väliset hitsi- ja ruuviliitokset siten, että rakenne hajoaa ylikuormitustilanteessa mahdollisimman turvallisesti. Työssä tehdyt laskelmat varmennettiin elementtimenetelmän avulla ja lopullisille rakenteille tehtiin elementtimenetelmällä vielä ominaistaajuusanalyysejä. Lopuksi työssä suunniteltiin runkorakenteille sopiva korroosionsuojaus.

Sähkökoneessa esiintyvien voimien yksityiskohtainen mittausjärjestelmä

Hissiteollisuudessa nostokoneistoina käytettyjen sähkömoottoreiden laatuvaatimukset ovat tiuken-tuneet viime vuosina. Erityisesti koneistojen tuottama ääni ja mekaaninen värähtely ovat olleet jat-kuvasti tiukentuneen tarkastelun alaisena. Hissikoriin ja hissiä ympäröiviin rakenteisiin välittyvästä värähtelystä johtuva ääni on yksi hissin laatuvaikutelmaan merkittävimmin vaikuttavia tekijöitä. Nostokoneisto on yksi tärkeimmistä äänen ja värähtelyn lähteistä hissijärjestelmässä. Koneiston suunnittelulla edellä mainittuja tekijöitä voidaan minimoida. Sähkökoneiden suunnittelussa finiit-tielementtimenetelmien (FEM) käyttö on vakiintunut haastavimmissa sovelluksissa. Kone Oyj:llä nostokoneistoina käytetään aksiaalivuokestomagneettitahtikoneita (AFPMSM), joiden FEM simu-lointiin käytetään yleisesti kolmea eri tapaa. Kukin näistä vaihtoehdoista pitää sisällään omat hyö-tynsä, että haittansa. Suunnittelun kannalta tärkeää on oikean menetelmän valinta ai-ka/informatiivisuus suhteen maksimoimiseksi. Erittäin tärkeää on myös saatujen tulosten oikeelli-suus. Tämän diplomityön tavoite on kehittää järjestelmä, jonka avulla AFPMS-koneen voimia voidaan mitata yksityiskohtaisella tasolla. Järjestelmän avulla voidaan tarkastella käytössä olevien FE-menetelmien tulosten oikeellisuutta sekä äänen että värähtelyn syntymekanismeja. Järjestelmän tarkoitus on myös syventää Kone Oyj tietotaitoa AFPMS-koneiden toiminnasta. Tässä työssä esitellään AFPMS-koneen epäideaalisuuksia, jotka voivat vaikuttaa mittajärjestelmän suunnitteluun. Myös koneen epäideaalisuuksiin lukeutuvaa ääntä on tarkasteltu tässä työssä. Jotta työn tavoitteiden mukaista FE-menetelmien vertailua ja tulosten oikeellisuuden tarkastelua voitai-siin tehdä, myös yleisimpiä AFPMS-koneen FE-menetelmiä tarkastellaan. Työn tuloksena on mittajärjestelmän suunnitelma, jonka avulla voidaan toteuttaa kuuden vapausas-teen voimamittaus jokaiselle koneistomagneetille alle 1N resoluutiolla. Suunnitellun järjestelmän toimivuutta on tarkasteltu FE-menetelmiä käyttäen ja järjestelmässä käytettävän voima-anturin ky-vykkyyttä on todennettu referenssimittauksin. Suunniteltu mittajärjestelmä mahdollistaa sähkömoottorin useiden eri epäideaalisuuksien tarkaste-lun yksityiskohtaisella tasolla. Mittausajatuksen soveltaminen myös muiden koneiden tutkimiseen tarjoaa mahdollisuuksia jatkotutkimuksille.

Global microRNA profiles and signaling pathways in the development of cardiac hypertrophy

Hypertrophy is a major predictor of progressive heart disease and has an adverse prognosis. MicroRNAs (miRNAs) that accumulate during the course of cardiac hypertrophy may participate in the process. However, the nature of any interaction between a hypertrophy-specific signaling pathway and aberrant expression of miRNAs remains unclear. In this study, Spague Dawley male rats were treated with transverse aortic constriction (TAC) surgery to mimic pathological hypertrophy. Hearts were isolated from TAC and sham operated rats (n=5 for each group at 5, 10, 15, and 20 days after surgery) for miRNA microarray assay. The miRNAs dysexpressed during hypertrophy were further analyzed using a combination of bioinformatics algorithms in order to predict possible targets. Increased expression of the target genes identified in diverse signaling pathways was also analyzed. Two sets of miRNAs were identified, showing different expression patterns during hypertrophy. Bioinformatics analysis suggested the miRNAs may regulate multiple hypertrophy-specific signaling pathways by targeting the member genes and the interaction of miRNA and mRNA might form a network that leads to cardiac hypertrophy. In addition, the multifold changes in several miRNAs suggested that upregulation of rno-miR-331*, rno-miR-3596b, rno-miR-3557-5p and downregulation of rno-miR-10a, miR-221, miR-190, miR-451 could be seen as biomarkers of prognosis in clinical therapy of heart failure. This study described, for the first time, a potential mechanism of cardiac hypertrophy involving multiple signaling pathways that control up- and downregulation of miRNAs. It represents a first step in the systematic discovery of miRNA function in cardiovascular hypertrophy.

Safety and outcome of coronary interventions with special reference to anticoagulation and stent type

University of Turku, Faculty of Medicine, Department of Cardiology and Cardiovascular Medicine, Doctoral Programme of Clinical Investigation, Heart Center, Turku University Hospital, Turku, Finland Division of Internal Medicine, Department of Cardiology, Seinäjoki Central Hospital, Seinäjoki, Finland Heart Center, Satakunta Central Hospital, Pori, Finland Annales Universitatis Turkuensis Painosalama Oy, Turku, Finland 2015 Antithrombotic therapy during and after coronary procedures always entails the challenging establishment of a balance between bleeding and thrombotic complications. It has been generally recommended to patients on long-term warfarin therapy to discontinue warfarin a few days prior to elective coronary angiography or intervention to prevent bleeding complications. Bridging therapy with heparin is recommended for patients at an increased risk of thromboembolism who require the interruption of anticoagulation for elective surgery or an invasive procedure. In study I, consecutive patients on warfarin therapy referred for diagnostic coronary angiography were compared to control patients with a similar disease presentation without warfarin. The strategy of performing coronary angiography during uninterrupted therapeutic warfarin anticoagulation appeared to be a relatively safe alternative to bridging therapy, if the international normalized ratio level was not on a supratherapeutic level. In-stent restenosis remains an important reason for failure of long-term success after a percutaneous coronary intervention (PCI). Drug-eluting stents (DES) reduce the problem of restenosis inherent to bare metal stents (BMS). However, a longer delay in arterial healing may extend the risk of stent thrombosis (ST) far beyond 30 days after the DES implantation. Early discontinuation of antiplatelet therapy has been the most important predisposing factor for ST. In study II, patients on long-term oral anticoagulant (OAC) underwent DES or BMS stenting with a median of 3.5 years’follow-up. The selective use of DESs with a short triple therapy seemed to be safe in OAC patients, since late STs were rare even without long clopidogrel treatment. Major bleeding and cardiac events were common in this patient group irrespective of stent type. In order to help to predict the bleeding risk in patients on OAC, several different bleeding risk scorings have been developed. Risk scoring systems have also been used also in the setting of patients undergoing a PCI. In study III, the predictive value of an outpatient bleeding risk index (OBRI) to identify patients at high risk of bleeding was analysed. The bleeding risk seemed not to modify periprocedural or long-term treatment choices in patients on OAC after a percutaneous coronary intervention. Patients with a high OBRI often had major bleeding episodes, and the OBRI may be suitable for risk evaluation in this patient group. Optical coherence tomography (OCT) is a novel technology for imaging intravascular coronary arteries. OCT is a light-based imaging modality that enables a 12–18 µm tissue axial resolution to visualize plaques in the vessel, possible dissections and thrombi as well as, stent strut appositions and coverage, and to measure the vessel lumen and lesions. In study IV, 30 days after titanium-nitride-oxide (TITANOX)-coated stent implantation, the binary stent strut coverage was satisfactory and the prevalence of malapposed struts was low as evaluated by OCT. Long-term clinical events in patients treated with (TITANOX)-coated bio-active stents (BAS) and paclitaxel-eluting stents (PES) in routine clinical practice were examined in study V. At the 3-year follow-up, BAS resulted in better long-term outcome when compared with PES with an infrequent need for target vessel revascularization. Keywords: anticoagulation, restenosis, thrombosis, bleeding, optical coherence tomography, titanium

Monolithic zirconium dioxide as a full contour restorative material: With special emphasis on the optical and mechanical properties

Full contour monolithic zirconia restorations have shown an increased popularity in the dental field over the recent years, owing to its mechanical and acceptable optical properties. However, many features of the restoration are yet to be researched and supported by clinical studies to confirm its place among the other indirect restorative materials This series of in vitro studies aimed at evaluating and comparing the optical and mechanical properties, light cure irradiance, and cement polymerization of multiple monolithic zirconia material at variable thicknesses, environments, treatments, and stabilization. Five different monolithic zirconia materials, four of which were partially stabilized and one fully stabilized were investigated. The optical properties in terms of surface gloss, translucency parameter, and contrast ratio were determined via a reflection spectrophotometer at variable thicknesses, coloring, sintering method, and after immersion in an acidic environment. Light cure irradiance and radiant exposure were quantified through the specimens at variable thicknesses and the degree of conversion of two dual-cure cements was determined via Fourier Transform Infrared spectroscopy. Bi-axial flexural strength was evaluated to compare between the partially and fully stabilized zirconia prepared using different coloring and sintering methods. Surface characterization was performed using a scanning electron microscope and a spinning disk confocal microscope. The surface gloss and translucency of the zirconia investigated were brand and thickness dependent with the translucency values decreasing as the thickness increased. Staining decreased the translucency of the zirconia and enhanced surface gloss as well as the flexural strength of the fully stabilized zirconia but had no effect on partially stabilized zirconia. Immersion in a corrosive acid increased surface gloss and decreased the translucency of some zirconia brands. Zirconia thickness was inversely related to the amount of light irradiance, radiant exposure, and degree of monomer conversion. Type of sintering furnace had no effect on the optical and mechanical properties of zirconia. Monolithic zirconia maybe classified as a semi-translucent material that is well influenced by the thickness, limiting its use in the esthetic zones. Conventional acid-base reaction, autopolymerizing and dual-cure cements are recommended for its cementation. Its desirable mechanical properties give it a high potential as a restoration for posterior teeth. However, close monitoring with controlled clinical studies must be determined before any definite clinical recommendations can be drawn.

Form, size and volumetric expansion of Adzuki beans (Vigna angularis) during soaking

This study analyzed the variation in shape and size of Adzuki beans during soaking at different temperatures. In addition, different mathematical models were fitted to the experimental values of volumetric expansion, selecting the best one. Grains of Adzuki beans (Vigna angularis) with moisture content of approximately 0.25 (decimal d.b.) were manually harvested; they were, then, dried to 0.128 (decimal d.b.). The beans were subjected to soaking in distilled water at the temperatures 18 ± 1, 27 ± 1, 36 ± 1, and 45 ± 1 °C, in five repetitions. Recipients containing 80 mL of distilled water and 20 g of beans for each sample were used. The samples were periodically weighed in order to determine the water absorption. After that, the samples were removed from the recipients and placed on filter papers for two minutes to drain the surface water. Water absorption continued until the beans reached the saturation moisture content. It was concluded that, the form of the Adzuki beans was altered regularly, the orthogonal axes expanded differentially in the radial and axial directions, and that the linear model appropriately described the volumetric expansion of the Adzuki beans, among the series of models analyzed for the temperatures 18, 27, 36 and 45 °C.

Biotechnological richness of the northeastern semi-arid region: antioxidant activity of casein hydrolysates from Moxotó goat milk (Capra hircus Linnaeus, 1758) obtained by papain action

This study aimed to identify antioxidant peptides from caprine casein hydrolysates by papain application using MALDI-TOF mass spectrometer, and a 2² full factorial design, with 4 axial points, in order to evaluate kinetic parameters (time and pH) effects on the degree of hydrolysis as well as the antioxidant activity of Moxotó goat milk casein peptides. Degree of hydrolysis was determined by total and soluble protein ratio in casein. Antioxidant activity was measured by ABTS method with 2, 2-cation-azinobis (3-ethylbenzothiazoline-6-sulfonic acid). TROLOX was used as standard. Peptide pattern and sequence of antioxidant amino acids were obtained using MALDI-TOF/MS. The highest degree of hydrolysis (28.5%) and antioxidant activity (2329.6 mmol.L TROLOX. mg- 1 peptide) were observed in the permeate. NENLL, NPWDQVK and LLYQEPVLGPV peptides, detected in the permeate, were pointed as the responsible for antioxidant activity, suggesting their potential application as food supplement and pharmaceutical products.

Eficácia, tolerabilidade e segurança do uso do sirolimo após o transplante renal

INTRODUÇÃO: Sirolimo (SRL) é um imunossupressor com conhecida eficácia e perfil de segurança na profilaxia da rejeição aguda após o transplante renal. OBJETIVOS: Avaliar eficácia, tolerabilidade e segurança do uso do SRL e de prednisona em associação a ciclosporina (CSA) ou tacrolimo (TAC) após o transplante renal. METODOLOGIA: Estudo retrospectivo de 332 receptores de transplantes renais realizados entre 1999 e 2006. O desfecho primário foi a falha de tratamento, definida como a incidência cumulativa de rejeição aguda confirmada por biópsia (RACB), perda do enxerto, óbito ou descontinuação do SRL. RESULTADOS: Dos 332 transplantes, 92% foram com doador vivo. A média de idade dos receptores foi de 37 anos, sendo 65% homens, 46% brancos e 6% diabéticos. SRL foi associado a CSA ou TAC em 70,8% e 29,2% dos pacientes. A incidência de falha de tratamento foi de 22,2% e de 47,8% no final do primeiro e do quinto ano de transplante, sem diferença entre pacientes recebendo CSA ou TAC. Ao final do quinto ano, as sobrevidas do paciente, do enxerto, do enxerto censorando o óbito e livre de RACB foram de 92,8%, 86,1%, 92,7% e 82,2%, respectivamente. O tratamento com SRL foi interrompido em 27,1% dos pacientes: 22,9% em razão de reações adversas e 3,3% devido à ineficácia. Os principais motivos de suspensão do SRL foram dislipidemia (6,0%), disfunção do enxerto (5,2%), proteinúria (4,5%), infecções (1,5%), dificuldade de cicatrização (1,2%) e anemia (0,9%). CONCLUSÃO: Na população estudada, a eficácia e a segurança do SRL foram semelhantes quando combinado com CSA ou TAC. A tolerabilidade oral foi adequada considerando-se a relativa baixa taxa de interrupção do uso de SRL.

Developing correlative Optical-Photoacustic microscopy for high-resolution in vivo imaging

PhotoAcoustic Imaging (PAI) is a branch in clinical and pre-clinical imaging, that refers to the techniques mapping acoustic signals caused by the absorption of the short laser pulse. This conversion of electromagnetic energy of the light to the mechanical (acoustic) energy is usually called photoacoustic effect. PAI, by combining optical excitation with acoustical detection, is able to preserve the diffraction limited spatial resolution. At the same time, the penetration depth is extended beyond the diffusive limit. The Laser-Scanning PhotoAcoustic Microscope system (LS-PAM) has been developed, that offers the axial resolution of 7.75 µm with the lateral resolution better than 10 µm. The first in vivo imaging experiments were carried out. Thus, in vivo label-free imaging of the mouse ear was performed. The principle possibility to image vessels located in deep layers of the mouse skin was shown. As well as that, a gold printing sample, vasculature of the Chick Chorioallantoic Membrane Assay, Drosophila larvae were imaged by PAI. During the experimental work, a totally new application of PAM was found, in which the acoustic waves, generated by incident light can be used for further imaging of another sample. In order to enhance the performance of the presented system two main recommendation can be offered. First, the current system should be transformed into reflection-mode setup system. Second, a more powerful source of light with the sufficient repetition rate should be introduced into the system.

Caracterização morfológica de frutos e sementes e desenvolvimento pós-seminal de monjoleiro (Acacia polyphylla DC.)

Acacia polyphylla DC. é uma espécie arbórea, característica dos estádios iniciais da sucessão secundária, de ocorrência natural no Brasil. Pertence à família Leguminosae-Mimosoideae, sendo recomendada em programas de reflorestamento misto, recuperação de áreas degradadas e manejo de fragmentos florestais. Entretanto, não foi encontrado nenhuma referência que fizesse menção aos aspectos químicos e morfológicos das sementes, bem como, aos aspectos do desenvolvimento pós-seminal desta espécie. Assim sendo, o presente trabalho objetivou caracterizar morfologicamente e ilustrar frutos e sementes, quantificar alguns componentes químicos presentes nas sementes e descrever as diferentes fases do desenvolvimento pós-seminal. Para tanto, foram realizadas descrições associadas às estruturas externa e interna das sementes. As descrições morfológicas dos frutos e das sementes foram efetuadas em relação a forma, ao tamanho, a superfície, a micrópila e a forma e a localização do embrião. Para a descrição morfológica das plântulas, as sementes foram colocadas para germinar em meio de cultura Murashige & Skoog reduzido à metade da concentração e incubadas a temperatura de 25ºC, sendo descritas e ilustradas as plântulas normais e anormais. O fruto é uma vagem deiscente contendo de oito a 16 sementes achatadas, de tegumento testal, embrião axial e invaginado. A germinação das sementes é epígea e as plântulas fanerocotiledonares.

Development of 3D super-resolution tomographic STED microscopy and its application to studies on bone resorption

In this doctoral thesis, a tomographic STED microscopy technique for 3D super-resolution imaging was developed and utilized to observebone remodeling processes. To improve upon existing methods, wehave used a tomographic approach using a commercially available stimulated emission depletion (STED) microscope. A certain region of interest (ROI) was observed at two oblique angles: one at a standard inverted configuration from below (bottom view) and another from the side (side view) via a micro-mirror positioned close to the ROI. The two viewing angles were reconstructed into a final tomogram. The technique, named as tomographic STED microscopy, was able to achieve an axial resolution of approximately 70 nm on microtubule structures in a fixed biological specimen. High resolution imaging of osteoclasts (OCs) that are actively resorbing bone was achieved by creating an optically transparent coating on a microscope coverglass that imitates a fractured bone surface. 2D super-resolution STED microscopy on the bone layer showed approximately 60 nm of lateral resolution on a resorption associated organelle allowing these structures to be imaged with super-resolution microscopy for the first time. The developed tomographic STED microscopy technique was further applied to study resorption mechanisms of OCs cultured on the bone coating. The technique revealed actin cytoskeleton with specific structures, comet-tails, some of which were facing upwards and some others were facing downwards. This, in our opinion, indicated that during bone resorption, an involvement of the actin cytoskeleton in vesicular exocytosis and endocytosis is present. The application of tomographic STED microscopy in bone biology demonstrated that 3D super-resolution techniques can provide new insights into biological 3D nano-structures that are beyond the diffraction-limit when the optical constraints of super-resolution imaging are carefully taken into account.