Tartrate-Resistant Acid Phosphatase 5b: a Serum Marker of Bone Resorption

Bone is a physiologically dynamic tissue being constantly regenerated throughout life as a consequence of bone turnover by bone-resorbing osteoclasts and bone-forming osteoblasts. In certain bone diseases, such as osteoporosis, the imbalance in bone turnover leads to bone loss and increased fracture risk. Measurement of bone mineral density (BMD) predicts the risk of fracture, but also biochemical markers of bone metabolism have been suggested to be suitable for prediction of fractures and monitoring the efficacy of antiresorptive treatment. Tartrate-resistant acid phosphatase 5b (TRACP 5b) is an enzyme released from osteoclasts into the circulation, from where it can be detected kinetically or immunologically. Conventional assays for serum total TRACP were spectrophotometric and suffered from interference by other acid phosphatases and non-osteoclastic TRACP 5a isoform. Our aim was to develop novel immunoassays for osteoclastic TRACP 5b. Serum TRACP 5b levels were elevated in individuals with high bone turnover, such as children, postmenopausal women, patients with osteoporosis, Paget’s disease and breast cancer patients with bone metastases. As expected, hormone replacement therapy (HRT) in postmenopausal women decreased the levels of serum TRACP 5b. Surprisingly, the highest TRACP 5b levels were observed in individuals with rare autosomal dominant osteopetrosis type II (ADO2), which is characterized by high BMD and fracture risk with simultaneously elevated levels of deficient osteoclasts. In ADO2 patients, elevated levels of serum TRACP 5b were associated with high fracture frequency. It is likely that serum TRACP 5b reflects the number of inactive osteoclasts in ADO2. Similar results supporting the hypothesis that TRACP 5b would reflect the number of osteoclasts instead of their activity were observed with cultured osteoclasts and in animal models. Novel TRACP 5b immunoassays may prove to be of value either as independent or combinatory tools with other bone metabolic markers and BMD measurements in clinical practice and bone research.

Osteoclastic tartrate-resistant acid phosphatase 5b. Diagnostic use and biological significance in bone physiology

The skeleton undergoes continuous turnover throughout life. In women, an increase in bone turnover is pronounced during childhood and puberty and after menopause. Bone turnover can be monitored by measuring biochemical markers of bone resorption and bone formation. Tartrate-resistant acid phosphatase (TRACP) is an enzyme secreted by osteoclasts, macrophages and dendritic cells. The secreted enzyme can be detected from the blood circulation by recently developed immunoassays. In blood circulation, the enzyme exists as two isoforms, TRACP 5a with an intact polypeptide chain and TRACP 5b in which the polypeptide chain consists of two subunits. The 5b form is predominantly secreted by osteoclasts and is thus associated with bone turnover. The secretion of TRACP 5b is not directly related to bone resorption; instead, the levels are shown to be proportional to the number of osteoclasts. Therefore, the combination of TRACP 5b and a marker reflecting bone degradation, such as C-terminal cross-linked telopeptides of type I collagen (CTX), enables a more profound analysis of the changes in bone turnover. In this study, recombinant TRACP 5a-like protein was proteolytically processed into TRACP 5b-like two subunit form. The 5b-like form was more active both as an acid phosphatase and in producing reactive oxygen species, suggesting a possible function for TRACP 5b in osteoclastic bone resorption. Even though both TRACP 5a and 5b were detected in osteoclasts, serum TRACP 5a levels demonstrated no change in response to alendronate treatment of postmenopausal women. However, TRACP 5b levels decreased substantially, demonstrating that alendronate decreases the number of osteoclasts. This was confirmed in human osteoclast cultures, showing that alendronate decreased the number of osteoclats by inducing osteoclast apoptosis, and TRACP 5b was not secreted as an active enzyme from the apoptotic osteoclasts. In peripubertal girls, the highest levels of TRACP 5b and other bone turnover markers were observed at the time of menarche, whereas at the same time the ratio of CTX to TRACP 5b was lowest, indicating the presence of a high number of osteoclasts with decreased resorptive activity. These results support the earlier findings that TRACP 5b is the predominant form of TRACP secreted by osteoclasts. The major source of circulating TRACP 5a remains to be established, but is most likely other cells of the macrophage-monocyte system. The results also suggest that bone turnover can be differentially affected by both osteoclast number and their resorptive activity, and provide further support for the possible clinical use of TRACP 5b as a marker of osteoclast number.

Physical activity in adolescence - with special reference to cardiovascular health

Liikunta ja sydän- ja verisuoniterveys nuorilla Aikuisiällä ilmenevien sydän- ja verisuonisairauksien kehitys alkaa lapsuudessa. Vähäinen liikunta lisää vaaraa sairastua sydän- ja verisuonisairauksiin sekä vaikuttaa haitallisesti näiden sairauksien riskitekijöihin läpi elämän. Tämän tutkimuksen tavoitteena oli tutkia nuorten liikuntaa ja sen yhteyttä sydän- ja verisuoniterveyteen. Sydän- ja verisuoniterveyttä tutkittiin riskitekijöiden sekä valtimon laajentumiskyvyn avulla. Tutkittavat koostuivat nuorista, jotka osallistuivat pitkäaikaisen sepelvaltimotaudin ehkäisytutkimuksen (STRIP) 13-vuotistutkimuskäynnille ja jotka raportoivat vapaa-ajan liikuntatottumuksensa (n=560). Liikunnan lisäksi nuorten pituus, paino, verenpaine ja olkavaltimon laajentumiskyky mitattiin sekä analysoitiin laboratorionäytteitä (otettiin verinäyte) ja selvitettiin ruoankäyttöä. Vähäinen vapaa-ajan liikunta oli yleistä etenkin tytöillä. Vapaa-ajallaan vähän liikkuvat pojat viettivät enemmän aikaa televisio- ja tietokoneruudun ääressä kuin vapaa-ajallaan paljon liikkuvat pojat. Äidin, toisin kuin isän, vapaa-ajan liikunta ja paino olivat yhteydessä lapsen vapaa-ajan liikuntaan. Vähän liikkuvat tytöt olivat olleet jo kahden vuoden iästä saakka useammin ylipainoisia kuin runsaammin liikkuvat ikätoverinsa. Sydän- ja verisuonisairauksien riskitekijöiden kasautuminen oli tavallisempaa vähän kuin paljon liikkuvilla nuorilla. Vähän liikkuvilla pojilla oli lisäksi huonompi olkavaltimon laajentumiskyky kuin paljon liikkuvilla pojilla. Tämä on tärkeä löydös, koska valtimon laajentumiskyky kuvannee sydän- ja verisuoniterveyttä jo ennen rakenteellisten muutosten ilmaantumista. Useat nuoret, etenkin tytöt, liikkuivat vähän vapaa-ajallaan. Vähäisellä vapaa-ajan liikunnalla oli haitallinen yhteys sydän- ja verisuonisairauksien riskitekijöihin sekä valtimon laajentumiskykyyn. Nuorten kannustaminen liikunnalliseen elämäntapaan on erittäin tärkeää sydän- ja verisuoniterveyden edistämiseksi sekä nuoren että kansanterveyden kannalta.

Imaging of the Vulnerable Atherosclerotic Plaque. Pre-Clinical Evaluation of PET Tracers for Vascular Inflammation

Atherosclerosis is a vascular inflammatory disease causing coronary artery disease, myocardial infarct and stroke, the leading causes of death in Finland and in many other countries. The development of atherosclerotic plaques starts already in childhood and is an ongoing process throughout life. Rupture of a plaque and the following occlusion of the vessel is the main reason for myocardial infarct and stroke, but despite extensive research, the prediction of rupture remains a major clinical problem. Inflammation is considered a key factor in the vulnerability of plaques to rupture. Measuring the inflammation in plaques non-invasively is one potential approach for identification of vulnerable plaques. The aim of this study was to evaluate tracers for positron emission tomography (PET) imaging of vascular inflammation. The studies were performed with a mouse model of atherosclerosis by using ex vivo biodistribution, autoradiography and in vivo PET and computed tomography (CT). Several tracers for inflammation activity were tested and compared with the morphology of the plaques. Inflammation in the atherosclerotic plaques was evaluated as expression of active macrophages. Systematic analysis revealed that the uptake of 18F-FDG and 11C-choline, tracers for metabolic activity in inflammatory cells, was more prominent in the atherosclerotic plaques than in the surrounding healthy vessel wall. The tracer for αvβ3 integrin, 18Fgalacto- RGD, was also found to have high potential for imaging inflammation in the plaques. While 11C-PK11195, a tracer targeted to receptors in active macrophages, was shown to accumulate in active plaques, the target-to-background ratio was not found to be ideal for in vivo imaging purposes. In conclusion, tracers for the imaging of inflammation in atherosclerotic plaques can be tested in experimental pre-clinical settings to select potential imaging agents for further clinical testing. 18F-FDG, 18F-galacto-RGD and 11C-choline choline have good properties, and further studies to clarify their applicability for atherosclerosis imaging in humans are warranted.

Neural correlates of language learning in adults

The human language-learning ability persists throughout life, indicating considerable flexibility at the cognitive and neural level. This ability spans from expanding the vocabulary in the mother tongue to acquisition of a new language with its lexicon and grammar. The present thesis consists of five studies that tap both of these aspects of adult language learning by using magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) during language processing and language learning tasks. The thesis shows that learning novel phonological word forms, either in the native tongue or when exposed to a foreign phonology, activates the brain in similar ways. The results also show that novel native words readily become integrated in the mental lexicon. Several studies in the thesis highlight the left temporal cortex as an important brain region in learning and accessing phonological forms. Incidental learning of foreign phonological word forms was reflected in functionally distinct temporal lobe areas that, respectively, reflected short-term memory processes and more stable learning that persisted to the next day. In a study where explicitly trained items were tracked for ten months, it was found that enhanced naming-related temporal and frontal activation one week after learning was predictive of good long-term memory. The results suggest that memory maintenance is an active process that depends on mechanisms of reconsolidation, and that these process vary considerably between individuals. The thesis put special emphasis on studying language learning in the context of language production. The neural foundation of language production has been studied considerably less than that of perceptive language, especially on the sentence level. A well-known paradigm in language production studies is picture naming, also used as a clinical tool in neuropsychology. This thesis shows that accessing the meaning and phonological form of a depicted object are subserved by different neural implementations. Moreover, a comparison between action and object naming from identical images indicated that the grammatical class of the retrieved word (verb, noun) is less important than the visual content of the image. In the present thesis, the picture naming was further modified into a novel paradigm in order to probe sentence-level speech production in a newly learned miniature language. Neural activity related to grammatical processing did not differ between the novel language and the mother tongue, but stronger neural activation for the novel language was observed during the planning of the upcoming output, likely related to more demanding lexical retrieval and short-term memory. In sum, the thesis aimed at examining language learning by combining different linguistic domains, such as phonology, semantics, and grammar, in a dynamic description of language processing in the human brain.

Tuning cell motility : roles of nestin and vimentin in cancer cell invasion and motility

The cytoskeleton is a key feature of both prokaryotic and eukaryotic cells. Itis comprised of three protein families, one of which is the intermediate filaments (IFs). Of these, the IFs are the largest and most diverse. The IFs are expressed throughout life, and are involved in the regulation of cell differentiation, homeostasis, ageing and pathogenesis. The IFs not only provide structural integrity to the cell, they are also involved in a range of cellular functions from organelle trafficking and cell migration to signalling transduction. The IFs are highly dynamic proteins, able to respond and adapt their network rapidly in response to intra- and extra- cellular cues. Consequently they interact with a whole host of cellular signalling proteins, regulating function, and activity, and cellular localisation. While the function of some of the better-known IFs such as the keratins is well studied, the understanding of the function of two IFs, nestin and vimentin, is poor. Nestin is well known as a marker of differentiation and is expressed in some cancers. In cancer, nestin is primarily described as is a promoter of cell motility, however, how it fulfils this role remains undefined. Vimentin too is expressed in cancer, and is known to promote cell motility and is used as a marker for epithelial to mesenchymal transition (EMT). It is only in the last decade that studies have addressed the role that vimentin plays in cell motility and EMT. This work provides novel insight into how the IFs, nestin and vimentin regulate cell motility and invasion. In particular we show that nestin regulates the cellular localisation and organisation of two key facilitators of cell migration, focal adhesion kinase and integrins. We identify nestin as a regulator of extracellular matrix degradation and integrin-mediated cell invasion. Two further studies address the specific regulation of vimentin by phosphorylation. A detailed characterisation study identified key phosphorylation sites on vimentin, which are critical for proper organisation of the vimentin network. Furthermore, we show that the bioactive sphingolipids are vimentin network regulators. Specifically, the sphingolipids induced RhoA kinasedependent (ROCK) phosphorylation at vimentin S71, which lead to filament reorganisation and inhibition of cell migration. Together these studies shed new light into the regulation of nestin and vimentin during cell motility.

Training executive functions in the laboratory and in real life : cognitive consequences of computer-based exercises and bilingualism

The question of the trainability of executive functions and the impact of such training on related cognitive skills has stirred considerable research interest. Despite a number of studies investigating this, the question has not yet been solved. The general aim of this thesis was to investigate two very different types of training of executive functions: laboratory-based computerized training (Studies I-III) and realworld training through bilingualism (Studies IV-V). Bilingualism as a kind of training of executive functions is based on the idea that managing two languages requires executive resources, and previous studies have suggested a bilingual advantage in executive functions. Three executive functions were studied in the present thesis: updating of working memory (WM) contents, inhibition of irrelevant information, and shifting between tasks and mental sets. Studies I-III investigated the effects of computer-based training of WM updating (Study I), inhibition (Study II), and set shifting (Study III) in healthy young adults. All studies showed increased performance on the trained task. More importantly, improvement on an untrained task tapping the trained executive function (near transfer) was seen in Study I and II. None of the three studies showed improvement on untrained tasks tapping some other cognitive function (far transfer) as a result of training. Study I also used PET to investigate the effects of WM updating training on a neurotransmitter closely linked to WM, namely dopamine. The PET results revealed increased striatal dopamine release during WM updating performance as a result of training. Study IV investigated the ability to inhibit task-irrelevant stimuli in bilinguals and monolinguals by using a dichotic listening task. The results showed that the bilinguals exceeded the monolinguals in inhibiting task-irrelevant information. Study V introduced a new, complementary research approach to study the bilingual executive advantage and its underlying mechanisms. To circumvent the methodological problems related to natural groups design, this approach focuses only on bilinguals and examines whether individual differences in bilingual behavior correlate with executive task performances. Using measures that tap the three above-entioned executive functions, the results suggested that more frequent language switching was associated with better set shifting skills, and earlier acquisition of the second language was related to better inhibition skills. In conclusion, the present behavioral results showed that computer-based training of executive functions can improve performance on the trained task and on closely related tasks, but does not yield a more general improvement of cognitive skills. Moreover, the functional neuroimaging results reveal that WM training modulates striatal dopaminergic function, speaking for training-induced neural plasticity in this important neurotransmitter system. With regard to bilingualism, the results provide further support to the idea that bilingualism can enhance executive functions. In addition, the new complementary research approach proposed here provides some clues as to which aspects of everyday bilingual behavior may be related to the advantage in executive functions in bilingual individuals.

Resa i röstens landskap : en narrativ studie av hur lärare blir professionella röstanvändare

The theme of this thesis is the learning process that occurs when teachers become professional voice users. The aim is to explore what it (really) means to become a professional voice user in a teaching profession; thereby developing an understanding of how future education in this field can be arranged so as to both effectively prevent vocal problems and to develop oral didactical competence among teachers. The ambition is to describe, interpret, and understand the learning process through a combination of emic and ethic research perspectives. The theoretical frame of reference reflects the cross disciplinary character of the thesis. Voice problems are common among both student teachers and inservice teachers and adversely affect professional competence, identity and quality of life. Additionally, vocal problems are proven to have a negative impact on pupils´ learning. The individual elements of learning are explored in the light of experiential learning theory and transformative learning theory. The social elements of learning are explored in relation to the theory of situated learning. In addition, theories of teacher professionalisation in terms of competence and identity are outlined. The empirical study has a longitudinal and multi method character. It is anchored in a phenomenological hermeneutical tradition, more specifically in narrative inquiry. The point of departure is the learning experiences of ten student teachers, who attended a ten week long course on voice production as part of their teacher training at Åbo Akademi University, in the autumn of 2002 and the spring of 2003. Four interviews in the form of conversations were conducted with each participant. These were crystallised with a process diary, a Swedish Voice Handicap Index, a voice observation, and a video observation. A fifth interview was conducted with each participant five years post teacher training, in the spring of 2008. Participant observation was also conducted throughout the course. The research materials have been analysed and interpreted narratively using a phenomenological hermeneutical method. The results are presented descriptively as individual narratives, which are reflected in logopedic research materials. Learning is here understood as emergent awareness. This is followed by a meta narrative concerning learning as experiences in the four dimensions body, thought, feeling, and relation. Finally, interpretation is expressed with respect to the theory of relational education. Learning is here understood as a movement in the field between the actual and the possible voice. It is also viewed as fundamentally rooted in inter-human relationships, in moments of presence and coexistence. As a tentative answer to the call for an existential space for learning in order to be a professional voice user, I suggest the concept of a learning refuge as a locus for a learning process built on trust, mutuality and openness.

The holistic military capability life cycle model

Abstract - This paper reviews existing military capability models and the capability life cycle. It proposes a holistic capability life-cycle model (HCLCM) that combines capability systems with related capability models. ISO 15288 standard is used as a framework to construct the HCLCM. The HCLCM also shows how capability models and systems relate to each other throughout the capability life cycle. The main contribution of this paper is conceptual in nature. The model complements the existing, but still evolving, understanding of the military capability life cycle in a holistic and systemic way. The model also increases understanding and facilitates communication among various military capability stakeholders.

Item-level life-cycle model for maintenance network – from cost to additional value

The importance of industrial maintenance has been emphasized during the last decades; it is no longer a mere cost item, but one of the mainstays of business. Market conditions have worsened lately, investments in production assets have decreased, and at the same time competition has changed from taking place between companies to competition between networks. Companies have focused on their core functions and outsourced support services, like maintenance, above all to decrease costs. This new phenomenon has led to increasing formation of business networks. As a result, a growing need for new kinds of tools for managing these networks effectively has arisen. Maintenance costs are usually a notable part of the life-cycle costs of an item, and it is important to be able to plan the future maintenance operations for the strategic period of the company or for the whole life-cycle period of the item. This thesis introduces an itemlevel life-cycle model (LCM) for industrial maintenance networks. The term item is used as a common definition for a part, a component, a piece of equipment etc. The constructed LCM is a working tool for a maintenance network (consisting of customer companies that buy maintenance services and various supplier companies). Each network member is able to input their own cost and profit data related to the maintenance services of one item. As a result, the model calculates the net present values of maintenance costs and profits and presents them from the points of view of all the network members. The thesis indicates that previous LCMs for calculating maintenance costs have often been very case-specific, suitable only for the item in question, and they have also been constructed for the needs of a single company, without the network perspective. The developed LCM is a proper tool for the decision making of maintenance services in the network environment; it enables analysing the past and making scenarios for the future, and offers choices between alternative maintenance operations. The LCM is also suitable for small companies in building active networks to offer outsourcing services for large companies. The research introduces also a five-step constructing process for designing a life-cycle costing model in the network environment. This five-step designing process defines model components and structure throughout the iteration and exploitation of user feedback. The same method can be followed to develop other models. The thesis contributes to the literature of value and value elements of maintenance services. It examines the value of maintenance services from the perspective of different maintenance network members and presents established value element lists for the customer and the service provider. These value element lists enable making value visible in the maintenance operations of a networked business. The LCM added with value thinking promotes the notion of maintenance from a “cost maker” towards a “value creator”.