Lymphatic vessels in health and disease: Role of the VEGF-C/VEGFR-3 pathway and the transcription factor FOXC2

The circulatory system consists of two vessel types, which act in concert but significantly differ from each other in several structural and functional aspects as well as in mechanisms governing their development. The blood vasculature transports oxygen, nutrients and cells to tissues whereas the lymphatic vessels collect extravasated fluid, macromolecules and cells of the immune system and return them back to the blood circulation. Understanding the molecular mechanisms behind the developmental and functional regulation of the lymphatic system long lagged behind that of the blood vasculature. Identification of several markers specific for the lymphatic endothelium, and the discovery of key factors controlling the development and function of the lymphatic vessels have greatly facilitated research in lymphatic biology over the past few years. Recognition of the crucial importance of lymphatic vessels in certain pathological conditions, most importantly in tumor metastasis, lymphedema and inflammation, has increased interest in this vessel type, for so long overshadowed by its blood vascular cousin. VEGF-C (Vascular Endothelial Growth Factor C) and its receptor VEGFR-3 are essential for the development and maintenance of embryonic lymphatic vasculature. Furthermore, VEGF-C has been shown to be upregulated in many tumors and its expression found to positively correlate with lymphatic metastasis. Mutations in the transcription factor FOXC2 result in lymphedema-distichiasis (LD), which suggests a role for FOXC2 in the regulation of lymphatic development or function. This study was undertaken to obtain more information about the role of the VEGF-C/VEGFR-3 pathway and FOXC2 in regulating lymphatic development, growth, function and survival in physiological as well as in pathological conditions. We found that the silk-like carboxyterminal propeptide is not necessary for the lymphangiogenic activity of VEGF-C, but enhances it, and that the aminoterminal propeptide mediates binding of VEGF-C to the neuropilin-2 coreceptor, which we suggest to be involved in VEGF-C signalling via VEGFR-3. Furthermore, we found that overexpression of VEGF-C increases tumor lymphangiogenesis and intralymphatic tumor growth, both of which could be inhibited by a soluble form of VEGFR-3. These results suggest that blocking VEGFR-3 signalling could be used for prevention of lymphatic tumor metastasis. This might prove to be a safe treatment method for human cancer patients, since inhibition of VEGFR-3 activity had no effect on the normal lymphatic vasculature in adult mice, though it did lead to regression of lymphatic vessels in the postnatal period. Interestingly, in contrast to VEGF-C, which induces lymphangiogenesis already during embryonic development, we found that the related VEGF-D promotes lymphatic vessel growth only after birth. These results suggest, that the lymphatic vasculature undergoes postnatal maturation, which renders it independent of ligand induced VEGFR-3 signalling for survival but responsive to VEGF-D for growth. Finally, we show that FOXC2 is necessary for the later stages of lymphatic development by regulating the morphogenesis of lymphatic valves, as well as interactions of the lymphatic endothelium with vascular mural cells, in which it cooperates with VEGFR-3. Furthermore, our study indicates that the absence of lymphatic valves, abnormal association of lymphatic capillaries with mural cells and an increased amount of basement membrane underlie the pathogenesis of LD. These findings have given new insight into the mechanisms of normal lymphatic development, as well as into the pathogenesis of diseases involving the lymphatic vasculature. They also reveal new therapeutic targets for the prevention and treatment of tumor metastasis and lymphatic vascular failure in certain forms of lymphedema. Several interesting questions were posed that still need to be addressed. Most importantly, the mechanism of VEGF-C promoted tumor metastasis and the molecular nature of the postnatal lymphatic vessel maturation remain to be elucidated.

VEGFR-2 and VEGFR-3 Specific Signaling in Lymphangiogenesis and Angiogenesis

The circulatory system consists of the blood and lymphatic vessels. While blood vessels transport oxygen, cells, and nutrients to tissues, the lymphatic vessels collect fluid, cells, and plasma proteins from tissues to return back to the blood circulation. Angiogenesis, the growth of new blood vessels from pre-existing ones, is an important process involved in several physiological conditions such as inflammation, wound healing, and embryonic development. Furthermore, angiogenesis is found in many pathological conditions such as atherosclerosis and the growth and differentiation of solid tumors. Many tumor types spread via lymphatic vessels to form lymph node metastasis. The elucidation of the molecular players coordinating development of the vascular system has provided an array of tools for further insight of the circulatory system. The discovery of the Vascular Endothelial Growth Factor (VEGF) family members and their tyrosine kinase receptors (VEGFRs) has facilitated the understanding of the vasculature in different physiological and pathological situations. The VEGFRs are expressed on endothelial cells and mediate the growth and maintenance of both the blood and lymphatic vasculatures. This study was undertaken to address the role of VEGFR-2 specific signaling in maturation of blood vessels during neoangiogenesis and in lymphangiogenesis. We also wanted to differentiate between VEGFR-2 and VEGFR-3 specific signaling in lymphangiogenesis. We found that specific VEGFR-2 stimulation alone by gene therapeutic methods is not sufficient for production of mature blood vessels. However, VEGFR-2 stimulation in combination with expression of platelet-derived growth factor D (PDGF-D), a recently identified member of the PDGF growth factor family, was capable of stabilizing these newly formed vessels. Signaling through VEGFR-3 is crucial during developmental lymphangiogenesis, but we showed that the lymphatic vasculature becomes independent of VEGFR-3 signaling after the postnatal period. We also found that VEGFR-2 specific stimulation cannot rescue the loss of lymphatic vessels when VEGFR-3 signaling is blocked and that VEGFR-2 specific signals promote lymphatic vessel enlargement, but are not involved in vessel sprouting to generate new lymphatic vessels in vivo, in contrast to the VEGFR-2 dependent sprouting observed in blood vessels. In addition, we compared the inhibitory effects of a small molecular tyrosine kinase inhibitor of VEGFR-2 vs. VEGFR-3 specific signaling in vitro and in vivo. Our results showed that the tyrosine kinase inhibitor could equally affect physiological and pathological processes dependent on VEGFR-2 and VEGFR-3 driven angiogenesis or lymphangiogenesis. These results provide new insights into the VEGFR specific pathways required for pre- and postnatal angiogenesis as well as lymphangiogenesis, which could provide important targets and therapies for treatment of diseases characterized by abnormal angiogenesis or lymphangiogenesis.

Dynamical processes in the stratosphere and upper troposphere and their influence on the distribution of trace gases in the polar atmosphere

Transport plays an important role in the distribution of long-lived gases such as ozone and water vapour in the atmosphere. Understanding of observed variability in these gases as well as prediction of the future changes depends therefore on our knowledge of the relevant atmospheric dynamics. This dissertation studies certain dynamical processes in the stratosphere and upper troposphere which influence the distribution of ozone and water vapour in the atmosphere. The planetary waves that originate in the troposphere drive the stratospheric circulation. They influence both the meridional transport of substances as well as parameters of the polar vortices. In turn, temperatures inside the polar vortices influence abundance of the Polar Stratospheric Clouds (PSC) and therefore the chemical ozone destruction. Wave forcing of the stratospheric circulation is not uniform during winter. The November-December averaged stratospheric eddy heat flux shows a significant anticorrelation with the January-February averaged eddy heat flux in the midlatitude stratosphere and troposphere. These intraseasonal variations are attributable to the internal stratospheric vacillations. In the period 1979-2002, the wave forcing exhibited a negative trend which was confined to the second half of winter only. In the period 1958-2002, area, strength and longevity of the Arctic polar vortices do not exhibit significant long-term changes while the area with temperatures lower than the threshold temperature for PSC formation shows statistically significant increase. However, the Arctic vortex parameters show significant decadal changes which are mirrored in the ozone variability. Monthly ozone tendencies in the Northern Hemisphere show significant correlations (|r|=0.7) with proxies of the stratospheric circulation. In the Antarctic, the springtime vortex in the lower stratosphere shows statistically significant trends in temperature, longevity and strength (but not in area) in the period 1979-2001. Analysis of the ozone and water vapour vertical distributions in the Arctic UTLS shows that layering below and above the tropopause is often associated with poleward Rossby wave-breaking. These observations together with calculations of cross-tropopause fluxes emphasize the importance of poleward Rossby wave breaking for the stratosphere-troposphere exchange in the Arctic.

Water in Action : Encounters among 2- to 3-Year-Old Children, Adults, and Water in Day Care

In line with cultural psychology and developmental theory, a single case approach is applied to construct knowledge on how children s interaction emerge interlinked to historical, social, cultural, and material context. The study focuses on the negotiation of constraints and meaning construction among 2-to 3-year-old children, a preschool teacher, and the researcher in settings with water. Water as an element offers a special case of cultural canalization: adults selectively monitor and guide children s access to it. The work follows the socio-cultural tradition in psychology, particularly the co-constructivist theory of human development and the Network of Meanings perspective developed at the University of São Paulo. Valsiner s concepts of Zone of Free Movement and Zone of Promoted Action are applied together with studies where interactions are seen as spaces of construction where negotiation of constraints for actions, emotions, and conceptions occur. The corpus was derived at a Finnish municipal day care centre. During a seven months period, children s actions were video recorded in small groups twice a month. The teacher and the researcher were present. Four sessions with two children were chosen for qualitative microanalysis; the analysis also addressed the transformations during the months covered by the study. Moreover, the data derivation was analyzed reflectively. The narrowed down arenas for actions were continuously negotiated among the participants both nonverbally and verbally. The adults expectations and intentions were materialized in the arrangements of the setting canalizing the possibilities for actions. Children s co-regulated actions emerged in relation to the adults presence, re-structuring attempts, and the constraints of the setting. Children co-constructed novel movements and meanings in relation to the initiatives and objects offered. Gestures, postures, and verbalizations emerged from the initially random movements and became constructed to have specific meanings and functions; meaning construction became abbreviated. The participants attempted to make sense of the ambiguous (explicit and implicit) intentions and fuzzy boundaries of promoted and possible actions: individualized yet overlapping features were continuously negotiated by all the participants. Throughout the months, children s actions increasingly corresponded adults (re-defined) conceptions of water researchers as an emerging group culture. Water became an instrument and a context for co-regulations. The study contributes to discussions on children as participants in cultural canalization and emphasizes the need for analysis in early childhood education practices on the implicit and explicit constraint structures for actions.

Exclusion of an Exotic Top Quark with -4/3 Electric Charge Using Soft Lepton Tagging

We present a measurement of the electric charge of the top quark using $\ppbar$ collisions corresponding to an integrated luminosity of 2.7~fb$^{-1}$ at the CDF II detector. We reconstruct $\ttbar$ events in the lepton+jets final state and use kinematic information to determine which $b$-jet is associated with the leptonically- or hadronically-decaying $t$-quark. Soft lepton taggers are used to determine the $b$-jet flavor. Along with the charge of the $W$ boson decay lepton, this information permits the reconstruction of the top quark's electric charge. Out of 45 reconstructed events with $2.4\pm0.8$ expected background events, 29 are reconstructed as $\ttbar$ with the standard model $+$2/3 charge, whereas 16 are reconstructed as $\ttbar$ with an exotic $-4/3$ charge. This is consistent with the standard model and excludes the exotic scenario at 95\% confidence level. This is the strongest exclusion of the exotic charge scenario and the first to use soft leptons for this purpose.

Monimenetelmäinen kuntoutuksen tutkimus. Artikkelikatsaus kuntoutuksen 3. valtakunnallisesta tutkimusseminaarista

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Omega-3-rasvahappojen käyttö nivelrikkoisten koirien hoidossa

Nivelrikko on koirilla yleinen sairaus. Sen hoitoon yleisesti käytettävien tulehduskipulääkkeiden rinnalle etsitään hoitomuotoja tulehduskipulääkkeiden pitkäaikaiskäytöstä johtuvien haittavaikutusten takia. Tutkimuksemme tavoite oli selvittää omega-3-rasvahappojen käytön hyötyä nivelrikkoisten koirien hoidossa. Tutkimuksen alkuhetkellä tutkimusta vastaavasta aiheesta koirilla ei ollut tehty. Ihmisillä reumatoidin artriitin hoidossa omega-3-rasvahapoilla on todettu olevan positiivisia vaikutuksia, joten siihen viitaten myös nivelrikkoisilla koirilla saattaisi olla hyötyä omega-3-rasvahappojen käytöstä. Omega-3-rasvahapoilla on todettu olevan positiivisia vaikutuksia useisiin tulehduksellisiin sairauksiin. Tutkimushypoteesimme oli, että kalaöljyä syövän ryhmän kohtisuora maksimaalinen voima askelvoimalevyllä kasvaisi ja ryhmän arvioitu kivuliaisuus laskisi tutkimuksen aikana. Tutkimamme tuote oli omega-3-rasvahapporavintolisä (Doils® Nivelet, Nutraceuticoils, Belgia). Tutkimus oli satunnaistettu, kaksoissokkoutettu ja kontrolloitu kliininen tutkimus. Tutkimukseen haettiin koiria ilmoitusten perusteella. Alkukyselyn ja seulontakäynnin perusteella tutkimuksen aloitti 77 koiraa. Seulontakäynnillä koirille tehtiin kliininen yleistutkimus, ortopedinen tutkimus, suppea neurologinen tutkimus, röntgenkuvaus, askelvoimalevymittaus sekä verinäytteenotto. Koirat kävivät varsinaisen tutkimuksen aikana kahdella lähikäynnillä. Molemmilla kerroilla koirille suoritettiin röntgenkuvausta lukuun ottamatta samat toimenpiteet kuin seulontakäynnillä. Tutkimuksen aikana omistajat täyttivät kyselykaavakkeita yhteensä seitsemän kertaa. Tutkimuksessa arvioitiin nivelrikkoisten koirien kipua sekä omistajan että eläinlääkärin arvioiden avulla sekä askelvoimalevyn avulla. Arviointimenetelmistä saatuja tuloksia olivat omistajan täyttämästä kyselykaavakkeesta kipuindeksin kyselyn tulokset, liikkumisvaikeuksista kertova VAS, elämänlaadusta kertova VAS, varakipulääkkeiden käyttö, vertailevat kysymykset, omistajan arvio valmisteen tehosta ja koiran ryhmästä, eläinlääkärin arvio sekä askelvoimalevyltä saadut kaksi muuttujaa (kohtisuora maksimaalinen voima ja impulssi). Tutkimustuloksissa havaittiin omega-3-rasvahappojen käytössä nivelrikkoisten koirien hoidossa pieni hyöty. Kalaöljyryhmän ja lumeryhmän välillä ei havaittu tilastollisesti merkitsevää eroa. Ryhmien sisällä havaittiin muutamia tilastollisesti merkitseviä muutoksia. Kipuindeksin muutos paremmaksi kalaöljyryhmässä tutkimuksen aikana oli vahvasti tilastollisesti merkitsevä muutos (p=0,002). Myös askelvoimalevyn kohtisuoran maksimaalisen voiman parantuminen oli vahvasti tilastollisesti merkitsevä muutos kalaöljyä syöneessä ryhmässä(p=0,001). Myös lumeryhmässä oli kipuindeksissä ja askelvoimalevyn kohtisuorassa maksimaalisessa voimassa havaittavissa tilastollisesti merkitsevä trendi (p=0,07 & p=0,059), mutta p-arvot eivät yltäneet tilastolliseen merkitsevyyteen. Suuremmalla otoskoolla näissä muuttujissa olisi voitu havaita joko tilastollisesti merkitseviä eroja ryhmien välillä tai sitten olisimme voineet osoittaa molemmissa ryhmissä tapahtuneen positiivisia eli parantavia muutoksia. Tutkimuksessa ei havaittu sivuvaikutuksia kalaöljyryhmässä. Tutkimustulokset todistivat tutkimushypoteesimme heikosti todeksi. Omega-3-rasvahappoja voisi siis hyvin käyttää koirilla, joille ei sovi kipulääkkeet.