Generation and Characterization of Knockout Mice and Analysis of Patient Material Demonstrates a Role for Tissue Inhibitor of Metalloproteinases 4 (Timp4) in the Cardiovascular System and Tumor Metastasis

This thesis focuses on tissue inhibitor of metalloproteinases 4 (TIMP4) which is the newest member of a small gene and protein family of four closely related endogenous inhibitors of extracellular matrix (ECM) degrading enzymes. Existing data on TIMP4 suggested that it exhibits a more restricted expression pattern than the other TIMPs with high expression levels in heart, brain, ovary and skeletal muscle. These observations and the fact that the ECM is of special importance to provide the cardiovascular system with structural strength combined with elasticity and distensibility, prompted the present molecular biologic investigation on TIMP4. In the first part of the study the murine Timp4 gene was cloned and characterized in detail. The structure of murine Timp4 genomic locus resembles that in other species and of the other Timps. The highest Timp4 expression was detected in heart, ovary and brain. As the expression pattern of Timp4 gives only limited information about its role in physiology and pathology, Timp4 knockout mice were generated next. The analysis of Timp4 knockout mice revealed that Timp4 deficiency has no obvious effect on the development, growth or fertility of mice. Therefore, Timp4 deficient mice were challenged using available cardiovascular models, i.e. experimental cardiac pressure overload and myocardial infarction. In the former model, Timp4 deficiency was found to be compensated by Timp2 overexpression, whereas in the myocardial infarct model, Timp4 deficiency resulted in increased mortality due to increased susceptibility for cardiac rupture. In the wound healing model, Timp4 deficiency was shown to result in transient retardation of re-epithelialization of cutaneous wounds. Melanoma tumor growth was similar in Timp4 deficient and control mice. Despite of this, lung metastasis of melanoma cells was significantly increased in Timp4 null mice. In an attempt to translate the current findings to patient material, TIMP4 expression was studied in human specimens representing different inflammatory cardiovascular pathologies, i.e. giant cell arteritis, atherosclerotic coronary arteries and heart allografts exhibiting signs of chronic rejection. The results showed that cardiovascular expression of TIMP4 is elevated particularly in areas exhibiting inflammation. The results of the present studies suggest that TIMP4 has a special role in the regulation of tissue repair processes in the heart, and also in healing wounds and metastases. Furthermore, evidence is provided suggesting the usefulness of TIMP4 as a novel systemic marker for vascular inflammation.

Perfil epidemiológico de acidentes com material biológico entre estudantes de medicina em um pronto-socorro cirúrgico

OBJETIVO: avaliar os acidentes com material biológico entre estudantes de medicina estagiando em um pronto-socorro de trauma e identificar as principais situações relacionadas, causas atribuídas e prevenção. MÉTODOS: estudo com abordagem quantitativa. Os dados foram coletados através de um questionário, aplicado via internet, contendo perguntas fechadas de escolha múltipla, referentes a acidentes com material biológico. A amostra obtida foi 100 estudantes. RESULTADOS: trinta e dois se acidentaram com materiais biológicos. As atividades de maior risco foram anestesia local (39,47%), sutura (18,42%) e recapeamento de agulha (15,79%). As principais vias de exposição ao material biológico foram contato com olho ou mucosa, com 34%, através de seringa com agulha com 45%. Após a contaminação, apenas 52% notificaram o acidente ao setor responsável. CONCLUSÃO: as principais causas de acidente encontradas e vias de exposição podem ser atribuídas a diversos fatores, como falta de treinamento e ao não uso de equipamentos de proteção individual. Ações preventivas e educativas são de extrema importância para diminuir a incidência dos acidentes com materiais biológicos e melhorar a conduta pós-exposição. É preciso entender as principais causas atribuídas e situações relacionadas a fim de implantar medidas gerais e eficazes.

Repair of segmental bone defects with fiber-reinforced composite: a study of material development and an animal model on rabbits

The Repair of segmental defects in load-bearing long bones is a challenging task because of the diversity of the load affecting the area; axial, bending, shearing and torsional forces all come together to test the stability/integrity of the bone. The natural biomechanical requirements for bone restorative materials include strength to withstand heavy loads, and adaptivity to conform into a biological environment without disturbing or damaging it. Fiber-reinforced composite (FRC) materials have shown promise, as metals and ceramics have been too rigid, and polymers alone are lacking in strength which is needed for restoration. The versatility of the fiber-reinforced composites also allows tailoring of the composite to meet the multitude of bone properties in the skeleton. The attachment and incorporation of a bone substitute to bone has been advanced by different surface modification methods. Most often this is achieved by the creation of surface texture, which allows bone growth, onto the substitute, creating a mechanical interlocking. Another method is to alter the chemical properties of the surface to create bonding with the bone – for example with a hydroxyapatite (HA) or a bioactive glass (BG) coating. A novel fiber-reinforced composite implant material with a porous surface was developed for bone substitution purposes in load-bearing applications. The material’s biomechanical properties were tailored with unidirectional fiber reinforcement to match the strength of cortical bone. To advance bone growth onto the material, an optimal surface porosity was created by a dissolution process, and an addition of bioactive glass to the material was explored. The effects of dissolution and orientation of the fiber reinforcement were also evaluated for bone-bonding purposes. The Biological response to the implant material was evaluated in a cell culture study to assure the safety of the materials combined. To test the material’s properties in a clinical setting, an animal model was used. A critical-size bone defect in a rabbit’s tibia was used to test the material in a load-bearing application, with short- and long-term follow-up, and a histological evaluation of the incorporation to the host bone. The biomechanical results of the study showed that the material is durable and the tailoring of the properties can be reproduced reliably. The Biological response - ex vivo - to the created surface structure favours the attachment and growth of bone cells, with the additional benefit of bioactive glass appearing on the surface. No toxic reactions to possible agents leaching from the material could be detected in the cell culture study when compared to a nontoxic control material. The mechanical interlocking was enhanced - as expected - with the porosity, whereas the reinforcing fibers protruding from the surface of the implant gave additional strength when tested in a bone-bonding model. Animal experiments verified that the material is capable of withstanding load-bearing conditions in prolonged use without breaking of the material or creating stress shielding effects to the host bone. A Histological examination verified the enhanced incorporation to host bone with an abundance of bone growth onto and over the material. This was achieved with minimal tissue reactions to a foreign body. An FRC implant with surface porosity displays potential in the field of reconstructive surgery, especially regarding large bone defects with high demands on strength and shape retention in load-bearing areas or flat bones such as facial / cranial bones. The benefits of modifying the strength of the material and adjusting the surface properties with fiber reinforcement and bone-bonding additives to meet the requirements of different bone qualities are still to be fully discovered.

Fatores Associados ao Material Insuficiente em Punção Aspirativa por Agulha Fina nos Nódulos Sólidos da Mama

Objetivo: analisar alguns fatores que possam estar associados à ocorrência de material insuficiente nos aspirados da punção aspirativa por agulha fina (PAAF). Pacientes e Métodos: foram estudadas 351 citologias de pacientes com nódulos sólidos da mama, submetidas a PAAF, como parte de sua investigação diagnóstica. As lâminas foram analisadas por um único citologista, que classificou os esfregaços como malignos, suspeitos, benignos ou material insuficiente para diagnóstico. Foram avaliados a idade da paciente, o tamanho do tumor, o estádio clínico, o Serviço, o dispositivo utilizado na punção e o tipo de lesão puncionada, de acordo com a histologia. A significância de cada variável em relação ao material insuficiente foi testada pelo c². Resultados: houve 67 esfregaços classificados como material insuficiente (19%). O tipo de dispositivo utilizado, o tamanho do tumor, o Serviço e o estádio clínico das lesões não se relacionaram à quantidade de material suficiente ou insuficiente. A idade da paciente e o tipo histológico influenciaram a taxa de material insuficiente, sendo que as pacientes abaixo de 50 anos tiveram uma taxa de 12%, comparada a 30% daquelas acima de 50 anos (p<0,03). Quanto à histologia, nas pacientes com alterações funcionais benignas da mama, a taxa de material insuficiente foi de 30%, seguida daquelas com carcinoma ductal infiltrante, 20%, e, no fibroadenoma, 12% (p<0,02). Conclusão: esse estudo mostrou uma taxa de material insuficiente relativamente alta (19%), a qual foi influenciada pela idade da paciente e pelo tipo histológico da lesão.

Abordagem citogenética e molecular em material de abortos espontâneos

OBJETIVOS: avaliar o desempenho da citogenética e das técnicas de hibridização in situ fluorescente (FISH) e reação em cadeia da polimerase (PCR) no estudo das aneuploidias cromossômicas numéricas e na determinação do sexo fetal em amostras de abortos espontâneos. MÉTODOS: duzentos e dezenove amostras de produtos de abortos espontâneos foram submetidas a estudo citogenético. Deste total, 40 amostras foram também submetidas à técnica de PCR-nested para a determinação do sexo fetal: 32 foram selecionadas devido à falha de crescimento no estudo citogenético e oito foram escolhidas ao acaso. Vinte amostras foram selecionadas para detecção de aneuploidias cromossômicas pela técnica de FISH, utilizando-se sondas para os cromossomos 13, 18, 21, X e Y: 13 casos foram submetidos a FISH devido à falha de crescimento no estudo citogenético e sete foram escolhidos ao acaso. Foi calculada a taxa de sucesso (obtenção de cariótipo) de cada técnica. Para comparação das taxas de sucesso foi utilizado o teste de chi2, sendo considerados significantes resultados com p<0,05. Foi avaliado o índice de acerto entre os resultados das amostras submetidas a mais de um exame, tomando-se como padrão-ouro o resultado do estudo citogenético. RESULTADOS: houve crescimento celular em 84,9% das amostras submetidas a análise citogenética. Em 51,1% dos casos foram encontradas alterações cromossômicas: 65,2% trissomias, 17,9% triploidias, 9,4% tetraploidias, 4,2% monossomia do cromossomo X e 1,1% trissomia dupla, tetrassomia e alteração estrutural. A trissomia mais freqüente foi a do cromossomo 16 (39%). FISH e PCR tiveram taxa de sucesso de 90%, não diferindo significativamente do exame citogenético. Em todos os casos submetidos a mais de um exame os resultados foram concordantes. Nas amostras com falha de crescimento celular no exame citogenético e submetidas a outra técnica, a PCR obteve sucesso em 87,5% e a FISH em 84,6%. CONCLUSÃO: o estudo citogenético de restos ovulares de abortamentos espontâneos teve elevada taxa de sucesso e evidenciou anomalias cromossômicas em mais da metade dos casos. As técnicas de biologia molecular (PCR-nested e FISH) complementaram o estudo citogenético e permitiram a obtenção de resultados seguros na detecção de alterações cromossômicas numéricas e na determinação do sexo fetal.

Infeccoes de transmissao vertical em material abortivo e sangue com enfase em Toxoplasmose gondii

OBJETIVO: Analisar os resultados sorológicos, anatomopatológicos e parasitológicos de material abortivo para infecções com risco de transmissão vertical, com ênfase na toxoplasmose. MÉTODOS: Foi realizado um estudo coorte-transversal tratando da prevalência das doenças infectoparasitárias. Participaram da pesquisa 105 mulheres que sofreram aborto espontâneo completo e/ou incompleto; elas foram entrevistadas por meio de um questionário, e foram coletadas amostras de sangue e material abortivo. Foram realizados testes imunológicos para toxoplasmose, doença de Chagas, rubéola, citomegalovírus e sífilis e análise anatomopatológica nos restos ovulares. RESULTADOS: 55% das mulheres tinham entre 20 e 30 anos de idade. A maioria (68%) apresentou idade gestacional entre a 7ª e a 14ª semanas. 54,3% das mulheres tinham o ensino médio completo ou incompleto. Pela análise da sorologia, a infecção com risco de transmissão vertical mais frequente foi o citomegalovírus (CMV) com 97,1% de positividade, e em seguida a rubéola, com 95,2%. A toxoplasmose teve um percentual de 54,3%, a doença de Chagas, de 1,9% e a sífilis, de 0,95%. A análise dos laudos de biópsia demonstrou que 63,1% apresentaram inflamação e 34%, ausência de inflamação. Das análises sorológica, anatomopatológica e parasitológica das 105 mulheres, 57 foram soropositivas para T. gondii, e nenhuma teve resultado positivo para a Reação em Cadeia da Polimerase (PCR) e para inoculação em camundongos. CONCLUSÕES: A prevalência de doenças com risco de transmissão congênita nas mulheres com abortamento espontâneo é importante, sendo necessárias pesquisas visando esclarecer a etiologia do aborto.

Summaries of lecture recordings used as learning material in blended learning

Julkaisumaa: 530 AN ANT Alankomaiden Antillit

Characterisation of Laser Beam and Paper Material Interaction

It is known already from 1970´s that laser beam is suitable for processing paper materials. In this thesis, term paper materials mean all wood-fibre based materials, like dried pulp, copy paper, newspaper, cardboard, corrugated board, tissue paper etc. Accordingly, laser processing in this thesis means all laser treatments resulting material removal, like cutting, partial cutting, marking, creasing, perforation etc. that can be used to process paper materials. Laser technology provides many advantages for processing of paper materials: non-contact method, freedom of processing geometry, reliable technology for non-stop production etc. Especially packaging industry is very promising area for laser processing applications. However, there are only few industrial laser processing applications worldwide even in beginning of 2010´s. One reason for small-scale use of lasers in paper material manufacturing is that there is a shortage of published research and scientific articles. Another problem, restraining the use of laser for processing of paper materials, is colouration of paper material i.e. the yellowish and/or greyish colour of cut edge appearing during cutting or after cutting. These are the main reasons for selecting the topic of this thesis to concern characterization of interaction of laser beam and paper materials. This study was carried out in Laboratory of Laser Processing at Lappeenranta University of Technology (Finland). Laser equipment used in this study was TRUMPF TLF 2700 carbon dioxide laser that produces a beam with wavelength of 10.6 μm with power range of 190-2500 W (laser power on work piece). Study of laser beam and paper material interaction was carried out by treating dried kraft pulp (grammage of 67 g m-2) with different laser power levels, focal plane postion settings and interaction times. Interaction between laser beam and dried kraft pulp was detected with different monitoring devices, i.e. spectrometer, pyrometer and active illumination imaging system. This way it was possible to create an input and output parameter diagram and to study the effects of input and output parameters in this thesis. When interaction phenomena are understood also process development can be carried out and even new innovations developed. Fulfilling the lack of information on interaction phenomena can assist in the way of lasers for wider use of technology in paper making and converting industry. It was concluded in this thesis that interaction of laser beam and paper material has two mechanisms that are dependent on focal plane position range. Assumed interaction mechanism B appears in range of average focal plane position of 3.4 mm and 2.4 mm and assumed interaction mechanism A in range of average focal plane position of 0.4 mm and -0.6 mm both in used experimental set up. Focal plane position 1.4 mm represents midzone of these two mechanisms. Holes during laser beam and paper material interaction are formed gradually: first small hole is formed to interaction area in the centre of laser beam cross-section and after that, as function of interaction time, hole expands, until interaction between laser beam and dried kraft pulp is ended. By the image analysis it can be seen that in beginning of laser beam and dried kraft pulp material interaction small holes off very good quality are formed. It is obvious that black colour and heat affected zone appear as function of interaction time. This reveals that there still are different interaction phases within interaction mechanisms A and B. These interaction phases appear as function of time and also as function of peak intensity of laser beam. Limit peak intensity is the value that divides interaction mechanism A and B from one-phase interaction into dual-phase interaction. So all peak intensity values under limit peak intensity belong to MAOM (interaction mechanism A one-phase mode) or to MBOM (interaction mechanism B onephase mode) and values over that belong to MADM (interaction mechanism A dual-phase mode) or to MBDM (interaction mechanism B dual-phase mode). Decomposition process of cellulose is evolution of hydrocarbons when temperature is between 380- 500°C. This means that long cellulose molecule is split into smaller volatile hydrocarbons in this temperature range. As temperature increases, decomposition process of cellulose molecule changes. In range of 700-900°C, cellulose molecule is mainly decomposed into H2 gas; this is why this range is called evolution of hydrogen. Interaction in this range starts (as in range of MAOM and MBOM), when a small good quality hole is formed. This is due to “direct evaporation” of pulp via decomposition process of evolution of hydrogen. And this can be seen can be seen in spectrometer as high intensity peak of yellow light (in range of 588-589 nm) which refers to temperature of ~1750ºC. Pyrometer does not detect this high intensity peak since it is not able to detect physical phase change from solid kraft pulp to gaseous compounds. As interaction time between laser beam and dried kraft pulp continues, hypothesis is that three auto ignition processes occurs. Auto ignition of substance is the lowest temperature in which it will spontaneously ignite in a normal atmosphere without an external source of ignition, such as a flame or spark. Three auto ignition processes appears in range of MADM and MBDM, namely: 1. temperature of auto ignition of hydrogen atom (H2) is 500ºC, 2. temperature of auto ignition of carbon monoxide molecule (CO) is 609ºC and 3. temperature of auto ignition of carbon atom (C) is 700ºC. These three auto ignition processes leads to formation of plasma plume which has strong emission of radiation in range of visible light. Formation of this plasma plume can be seen as increase of intensity in wavelength range of ~475-652 nm. Pyrometer shows maximum temperature just after this ignition. This plasma plume is assumed to scatter laser beam so that it interacts with larger area of dried kraft pulp than what is actual area of beam cross-section. This assumed scattering reduces also peak intensity. So result shows that assumably scattered light with low peak intensity is interacting with large area of hole edges and due to low peak intensity this interaction happens in low temperature. So interaction between laser beam and dried kraft pulp turns from evolution of hydrogen to evolution of hydrocarbons. This leads to black colour of hole edges.

Punção aspirativa por agulha fina como método de coleta de material para a histopatologia no osteossarcoma canino

Para a obtenção do diagnóstico definitivo do osteossarcoma realizam-se exames citopatológico e histopatológico. O material para exame citopatológico é coletado através de punção aspirativa por agulha fina (PAAF), já para a realização do exame histopatológico é necessário uma amostra de tamanho maior, geralmente conseguida através de biópsia incisional. Este trabalho tem como objetivo desenvolver uma técnica de coleta de material em cães com suspeita de osteossarcoma através de PAAF para a realização de exame histopatológico. Foram coletadas duas amostras de 12 cães suspeitos de osteossarcoma por PAAF. O material obtido pela primeira coleta foi utilizado para confirmar o diagnóstico através do exame citopatológico, enquanto que o material oriundo da segunda coleta foi fixado em formol a 10% para a análise histopatológica. Quatro das 12 amostras (33,3%) avaliadas histopatologicamente pela metodologia proposta obtiveram também o diagnóstico de osteossarcoma. Esses resultados apontam para uma possível adequação da técnica de coleta de material por PAAF para exame histopatológico.

Carbon Nanotubes as Material for Supercapacitor Electrodes

Electrochemical double-layer supercapacitors have an intermediate position between rechargeable batteries, which can store high amounts of energy, and dielectric capacitors, which have high output power. Supercapacitors are widely suggested to be used in automobiles (recuperation during braking, facilitate engine starting, electric stabilization of the system), industry (forklifts, elevators), hybrid off-road machinery and also in consumer electronics. Supercapacitor electrodes require highly porous material. Typically, activated carbon is used. Specific surface area of activated carbon is approximately 1000 m2 per gram. Carbon nanotubes represent one of prospective materials. According to numerous studies this material allows to improve the properties of supercapacitors. The task of this Master‘s Thesis was to test multiwalled carbon nanotubes and become confident with the testing methods.

Value added logistics in supply and demand chain : COSTFIX : New operation model for maintenance material logistics in paper industry

This research report illustrates and examines new operation models for decreasing fixed costs and transforming them into variable costs in the field of paper industry. The report illustrates two cases – a new operation model for material logistics in maintenance and an examination of forklift truck fleet outsourcing solutions. Conventional material logistics in maintenance operation is illustrated and some problems related to conventional operation are identified. A new operation model that solves some of these problems is presented including descriptions of procurement and service contracts and sources of added value. Forklift truck fleet outsourcing solutions are examined by illustrating the responsibilities of a host company and a service provider both before and after outsourcing. The customer buys outsourcing services in order to improve its investment productivity. The mechanism of how these services affect the customer company’s investment productivity is illustrated.

Critical aspects on the behavior of material from the mechanical tool-workpiece interaction in single point diamond turning

Some material aspects such as grain size, purity and anisotropy exert an important influence on surface quality, especially in single point diamond turning. The aim of this paper is to present and discuss some critical factors that can limit the accuracy of ultraprecision machining of non-ferrous metals and to identify the effects of them on the cutting mechanism with single point diamond tools. This will be carried out through observations of machined surfaces and chips produced using optical and scanning electron microscopy. Solutions to reduce the influence of some of these limiting factors related with the mechanism of generation of mirror-like surfaces will be discussed.

On the development of an unstructured grid solver for inert and reactive high speed flow simulations

An unstructured grid Euler solver for reactive compressible flow applications is presented. The method is implemented in a cell centered, finite volume context for unstructured triangular grids. Three different schemes for spatial discretization are implemented and analyzed. Time march is implemented in a time-split fashion with independent integrators for the flow and chemistry equations. The capability implemented is tested for inert flows in a hypersonic inlet and for inert and reactive supersonic flows over a 2-D wedge. The results of the different schemes are compared with each other and with independent calculations using a structured grid code. The strengths and the possible weaknesses of the proposed methods are discussed.

Potential benefits of building information modeling for building material supplier

This thesis investigated building information modeling (BIM) from a material supplier’s point of view. The objective was to gain understanding about how a building material supplier could benefit from the growing use of BIM in the AEC (architectural, engineering and construction) industry. Increasing amount of inquiries related to BIM from customers and other interest groups had awoken target company’s interest towards BIM. This thesis acts as a pre-study for the target company related to potential of BIM. First of all BIM and its meaning from a material supplier’s point of view was defined based on a literature review. To reveal the potential benefits of BIM for a material supplier a questionnaire survey and in total of 11 interviews were conducted. Based on the literature review and analyzed results it came clear that BIM offers benefits also for material suppliers. Product libraries and material databases for BIM tools can act as an important marketing channel for material suppliers. Material suppliers could also utilize the information from the BIM models to schedule their deliveries more precisely and potentially even to schedule their own production. All this needs deeper cooperation between material suppliers, contractors and other stakeholders in the AEC industry. Based on the results also first steps for the target company to utilize the growing use of BIM were defined.