Novel Functions of ErbB4 and NRG-1 in Development and Cancer

Cells communicate, or signal, with each other constantly to ensure proper functioning of tissues and organs. Cell signaling is often performed by interplay of receptors and ligands that bind these receptors. ErbB receptors (epidermal growth factor receptors, EGFR, HER) bind extracellular growth factors and transduce these signals inside of cells. ErbB dysfunction promotes carcinogenesis, and also results in numerous defects during normal development. This study focused on the functions of one member of the ErbB receptor family, ErbB4, and growth factor, neuregulin-1 (NRG-1), that can bind and activate ErbB4. This study aimed to find novel functions of ErbB4 and NRG-1. Hypoxia, or deficiency of oxygen, is common in cancer and ischemic conditions. One of the key findings of the work was the identification and characterization of a cross-talk between ErbB4 and Hypoxia-inducible factor 1�� (HIF-1��), the central mediator of hypoxia signaling. ErbB4 activation by NRG-1 was found to increase HIF-1�� activity. Interestingly, this regulation occurred in reciprocal manner as HIF-1�� was also able to increase protein levels of NRG-1 and ErbB4. Moreover, expression of NRG-1 and ErbB4 was associated with HIF activity in vivo in human clinical samples and in mice. Reduction of functional ErbB4 in developing zebrafish embryos resulted in defects in development of the skeletal muscles. To study ErbB4 functions in pathological situation in humans, clinical samples of serous ovarian carcinoma were analyzed using tissue microarrays and real-time RT-PCR. A specific isoform of ErbB4, CYT-1, was associated with poor survival in serous ovarian cancer and increased anchorage independent growth of ovarian cancer cells in vitro. These observations demonstrate that ErbB4 and NRG-1 are essential regulators of cellular response to hypoxia, of development, and of ovarian carcinogenesis.

VAP-1 in Leukocyte Trafficking

The extravasation of leukocytes from the blood stream into the tissues is a prerequisite for adequate immune surveillance and immune reaction. The leukocyte movement from the bloodstream into the tissues is mediated by molecular bonds. The bonds are formed between adhesion molecules on endothelial cells and their counterparts expressed on leukocytes. Vascular adhesion protein-1 (VAP-1) is an endothelial adhesion molecule mediating leukocyte interactions with endothelium. It is also an enzyme having semicarbazide sensitive amine oxidase (SSAO) activity. The SSAOactivity catalyses deamination of primary amines into corresponding aldehyde and during the enzymatic reaction hydrogen peroxide and ammonia are produced. The aim of this study was to investigate the relationship between the adhesive and enzymatic activities of VAP-1. The role of VAP-1 in leukocyte traffic was studied <i>in vivo</i> under normal and pathological conditions in VAP-1 deficient mice. The results from <i>in vitro</i> flow-based assays indicated that VAP-1 uses both SSAOactivity and its adhesive epitope to bind leukocytes, and both are perquisites for VAP-1 mediated adhesion. Furthermore, <i>in vivo</i> results demonstrated that leukocyte trafficking was impaired <i>in vivo</i> by deleting VAP-1 or inhibiting SSAO-activity. There was impairment in lymphocyte recirculation as well as leukocyte accumulation into the inflamed area. Moreover, the VAP-1 deficient mice did not show generalized defects in antimicrobial responses, whereas significant reduction in tumor progression and neovascularization was observed. These results indicate that VAP-1 could be used as a target in anti-adhesive therapies either by blocking its adhesive epitope with antibodies or by inhibiting its SSAO-activity using inhibitors. Moreover, targeting of VAP-1 may provide a new way of inhibiting neovascularization in tumors.

��1 integrin regulation

Integrins are heterodimeric adhesion receptors mediating adhesion to extracellular matrix proteins and to other cells. Integrins are important in embryonic development, structural integrity of connective tissue, blood thrombus formation, and immune defense system. Integrins are transmembrane proteins whose ligand binding capacity (activity) is regulated by large conformational changes. Extracellular ligand binding or intracellular effector binding to integrin cytoplasmic face regulate integrin activity. Integrins are thus able to mediate bi-directional signaling. Integrin function is also regulated by intracellular location. Integrins are constantly recycled from endocytic vesicles to plasma membrane, and this has been shown to be important for cell migration and invasion as well. Deregulation of integrin functionality can lead to deleterious illnesses, such as bleeding or inflammatory disorders. It is also evident that integrin deregulation is associated with cancer progression. In this study, a novel Beta1 integrin associating protein, Rab21, was characterized. Rab21 binding to integrin cytoplasmic tail was shown to be important for Beta1 integrin endo- and exocytosis ��� intracellular trafficking. It was furher shown that this interaction has an important role in cell adhesion, migration, as well as in the final step of cell division, cytokinesis. This work showed that abrogation of Rab21 function or ��1 integrin endocytic traffic, can lead to defects in cell division and results in formation of multinucleated cells. Multinucleation and especially tetraploidy can be a transient pathway to aneuploidy and tumorigenesis. This work characterized chromosomal deletions in <i>rab21</i> locus in ovarian and prostate cancer samples and showed that a cell line with <i>rab21</i> deletion also had impairment in cell division, which could be rescued by Rab21 re-expression. The work demonstrates an important role for Rab21 and Beta1 integrin traffic regulation in cell adhesion and division, and suggests a probable associaton with tumorigenesis. In this study, Beta1 integrin activity regulation was also addressed. A novel cell array platform for genome-scale RNAi screenings was characterized here. More than 4500 genes were knocked-down in prostate cancer cells using siRNA-mediated silencing. The effects on Beta1 integrin activity were analyzed upon knock-downs. The screen identified more that 400 putative regulators of Beta1 integrin activity in prostate cancer. In conclusion, this work will help us to understand complex regulatory pathways involved in cancer cell adhesion and migration.

PV-1: Leukocyte trafficking molecule and vascular marker

The distinction between lymphatic vessels and blood vessels is a crucial factor in many studies in immunology, vascular biology and cancer biology. They both share several characteristics and perform related, though different functions. They are equally important for the performance of the human immune system with the continuous recirculation of leukocytes from the tissues via lymphatics to the blood vessels and back into the tissue presenting the link between both systems. This study was undertaken to elucidate the differences in the gene expression between primary blood- and lymphatic endothelial cells as well as the two immortalized cell lines HMEC-1 (human microvascular endothelial cell line 1) and TIME (telomerase immortalized microvascular endothelial cell line). Furthermore, we wanted to investigate the mystery surrounding the identity of the antigen recognized by the prototype blood vascular marker PAL-E. In the last step we wanted to study whether the PAL-E antigen would be involved in the process of leukocyte migration from the bloodstream into the surrounding tissue. Our results clearly show that the gene expression in primary blood endothelial cells (BEC), lymphatic endothelial cells (LEC) and the cell lines HMEC-1 and TIME is plastic. Comparison of a large set of BEC- and LEC datasets allowed us to assemble a catalog of new, stable BEC- or LEC specific markers, which we verified in independent experiments. Additionally, several lines of evidence demonstrated that PAL-E recognizes plasmalemma vesicle associated protein 1 (PV-1), which can form complexes with vimentin and neuropilin-1. Finally, numerous in vitro and in vivo experiments identify the first function of the protein PV-1 during leukocyte trafficking, where it acts as regulatory molecule.

CLEVER-1 as an immune suppressive molecule

The immune response and immune suppression are equally essential for the immune system to protect the host against an infection and to protect self-molecules in different pathophysiological conditions. Pregnancy is one of the conditions where the maternal immune system remains resistant against microbes and yet attains tolerance to protect the fetus, whose genetic material differs partially from the mother���s. However, if the balance of immune suppression is not precise in the host it can favor conditions which lead to diseases, such as cancer and autoimmune disorders. This study was initiated to investigate the expression and functions of CLEVER-1/Stabilin-1, a multifunctional protein expressed on subsets of endothelial cells and type II macrophages, as an immune suppressive molecule. Firstly, the expression of CLEVER-1/stabilin-1 and its function in human placental macrophages were examined. Secondly, the expression profile and functional significance of stabilin-1 on healthy human monocytes was investigated. The results clarified the expression of CLEVER-1/stabilin-1 on placental macrophages, and verified that CLEVER-1/stabilin-1 functions as an adhesion and scavenging molecule on these cells. The data from normal monocytes revealed that the monocytes with low stabilin-1 expression carried a pro-inflammatory gene signature, and that stabilin-1 can directly or indirectly regulate pro-inflammatory genes in monocytes. Finally, it was shown that monocyte CLEVER-1/stabilin-1 dampens IFN��� production by T cells. To conclude, CLEVER-1/stabilin-1 is defined as an immune suppressive molecule on monocytes and macrophages. Strikingly, anti-stabilin-1 antibodies may have the potential to promote the Th1 dependent inflammatory response and counteract the tumor induced immune suppression.

Vitamin B12 Deficiency in the aged: Laboratory Diagnosis, Prevalence and Clinical Profile

<B>B₁₂-vitamiinin puute i��kk��ill��: laboratoriodiagnostiikka, yleisyys ja yhteys sairastavuuteen</b> <b>Tausta:</b> B12-vitamiinin puute on yleist�� i��kk��ill�� ja se tulisi todeta riitt��v��n varhaisessa vaiheessa palautumattomien vaurioiden est��miseksi. On ep��selv���� pit��isik�� diagnostiikka kohdistaa tiettyihin riskiryhmiin vai mahdollisesti seuloa valikoimatonta vanhusv��est����. My��sk����n yksimielisyytt�� laboratoriotutkimusten valinnasta ei ole. <b>Tavoitteet: </b>Tutkimuksen tarkoituksena oli evaluoida uutta HoloTC RIA menetelm���� ja tuottaa viitearvot sille, selvitt���� B12-vitamiinin puutteen yleisyys, yhteys sairastavuuteen ja mahdolliset riskitekij��t suomalaisessa vanhusv��est��ss��, arvioida munuaisfunktion vaikusta B12-vitamiinin puutteen laboratoriotutkimuksiin ja n��iden perusteella ehdottaa suomalaiseen terveydenhuoltoon sopivaa laboratoriotutkimusstrategiaa. <b>Aineisto ja menetelm��t:</b> Liedon i��kk����t -tutkimuksen vanhusaineisto on edustava otos yhden kunnan yli 65-vuotiaasta v��est��st��, yhteens�� 1260 henkil����. Tutkittavat k��viv��t l����k��rintarkastuksessa, ja heist�� on k��ytett��viss�� runsaasti laboratoriotutkimuksia sek�� tiedot sairauksista, ruokavaliosta, l����kkeiden ja vitamiinivalmisteiden k��yt��st��, dementiaseula ja depressiokysely. Viitearvoaineistoa varten ker��ttiin n��ytteet 84 vapaaehtoisesta terveest�� aikuisesta ja menetelm��evaluaatiota varten 107 sairaalapotilaasta. <b>Tulokset:</b> HoloTC RIA menetelm��n toistettavuus oli hyv�� manuaalimenetelm��ksi. 95%:n viitev��li holotranskobalamiinille oli 37-171 pmol/l. Kaikilla tutkittavilla, joilla oli muilla laboratoriotutkimuksilla osoitettu todenn��k��inen B12-vitamiinin puute, my��s holotranskobalamiini oli viitealueen alarajaa pienempi. Suurentuneella kystatiini C-pitoisuudella osoitettu munuaisten vajaatoiminta korreloi voimakkaasti homokysteiinin (rs=0.53, p<0.001) ja metyylimalonihapon (rs=0.27, p<0.001) pitoisuuksiin, mutta ei kokonais-B12-vitamiinin (rs=- 0.04, p=0.227) tai holotranskobamiinin (rs=-0.01, p=0.817) pitoisuuksiin. Suomalaisessa vanhusv��est��ss�� B12-vitamiinin puutteen prevalenssi oli 12%. Kokonais- B12-vitamiinin pitoisuus oli matala (<150 pmol/l) 6%:lla. Miessukupuoli (OR 1.9, 95% CI 1.2-2.9), ik�� ���75 (OR 2.2, 95% CI 1.4-3.4) ja maitotuotteiden v��ltt��minen (OR 2.3, 95% CI 1.2-4.4) lis��siv��t B12-vitamiinin puutteen riski��, mutta anemia (OR 1.3, 95% CI 0.7-2.3) tai makrosytoosi (OR 1.2, 95% CI 0.6-2.7) eiv��t. <b>P����telm��t: </b>Diagnosoimaton B12-vitamiinin puute on yleist�� i��kk��ill��, mutta kliinisesti merkityksellist�� spesifist�� riskiryhm���� ei l��ydy. Koska anemian ja makrosytoosin puuttuminen ei poissulje B12-vitamiinin puutetta ja munuaisten vajaatoiminta heikent���� metabolisten merkkiaineiden k��ytt��kelpoisuutta, kokonais-B12-vitamiinia suositellaan ensisijaiseksi laboratoriotutkimukseksi ep��ilt��ess�� B12-vitamiinin puutetta ja tarvittaessa varmentavina tutkimuksina k��ytet����n homokysteiini�� ja holotranskobalamiinia.

Identiteetti ja luopuminen perheyrityksen sukupolvenvaihdoksessa

Tutkimus on perheyritt��jyystutkimusta, se k��sittelee perheyrityksess�� tapahtunutta sukupolvenvaihdosta sek�� siihen liittyv���� luopumisen prosessia. Poikkitieteellisen�� l��hestymistapana k��ytet����n sosiologisia ja psykologisia katsantokantoja identiteetin rakentumisesta ja sen muuttumisesta. Luopumisk��sitett�� l��hestyt����n perheyritt��j��n identiteetin rakentumisen ja muuttumisen kautta. Tutkimus etsii vastausta nelj����n kysymykseen: 1. Millainen on perheyritt��j��n identiteetti? 2. Miten perheyrityksen sukupolvenvaihdos vaikuttaa identiteettiin? 3. Mit�� luopuminen tarkoittaa sukupolvenvaihdoksessa? 4. Mit�� sukupolvenvaihdoksessa todellisuudessa tapahtuu? Tutkimuksessa paneudutaan luopujan ja jatkajan n��k��kulmiin, joita tarkastellaan el��m��nkertatarinoiden avulla. Tutkimuksen aineisto koostuu viidest�� yrityksest��, joissa jollakin tasolla on sukupolvenvaihdos toteutettu. Tutkimuksessa on haastateltu kolmeatoista henkil����. Tutkimuksen perusteella voidaan todeta, ett�� perheyritt��j��n identiteetti on hyvin kompleksinen ja vaikeasti l��hestytt��v�� asia. Perheyritt��j��n persoona on vahva ja perhe sek�� yritys vaikuttavat oleellisesti sen muotoutumiseen. Individuaalisuus j���� usein familistisen n��kemyksen jalkoihin. Sukupolvenvaihdos vaikutti sek�� luopujan ett�� jatkajan identiteettiin ratkaisevasti. Luopujat kohtasivat kriisin miettiess����n yrityksens�� tulevaisuutta ja moni jatkaja puolestaan muutti el��m����ns�� radikaalisti siirtyess����n perheyrityksen jatkajaksi. Ty��st�� luopuminen osoittautui tutkimuksessa kriittisimm��ksi tekij��ksi, etenkin luopujien n��k��kulmasta katsottuna. Jatkajille se merkitsi oman jo mahdollisen muun uran ja ty��paikan vaihtumista perheyritykseen. Luopujille se teoriassa tarkoitti el��kkeelle siirtymist��. Tutkituissa yrityksiss�� kukaan luopujista ei kuitenkaan ole lopettanut ty��n tekoa. He ovat jokap��iv��inen n��ky yrityksess�� ja enemm��n tai v��hemm��n aktiivisesti mukana yrityksen toiminnoissa. T��m�� aiheuttaa sukupolvien yhteent��rm��yst�� ja konflikteja, t��m�� seikka vie paljon energiaa yrityksess�� teht��v��lt�� ty��lt��. Ty�� on yritt��j��lle eritt��in vahva ja merkitsev�� identiteetin perusta, jonka muuttaminen tai siit�� luopuminen on vaikeaa.

The use of dietary supplements and medication among Finnish elite athletes

Background: Dietary supplements are widely used among elite athletes but the prevalence of dietary supplement use among Finnish elite athletes is largely not known. The use of asthma medication is common among athletes. In 2009, the World Anti-Doping Agency (WADA) and the International Olympic Committee (IOC) removed the need to document asthma by lung function tests before the use of inhaled ��2-agonists. Data about medication use by Paralympic athletes (PA) is limited to a study conducted at the Athens Paralympics. Aims: To investigate the prevalence of the use of self-reported dietary supplements, the use of physician-prescribed medication and the prevalence of physician-diagnosed asthma and allergies among Finnish Olympic athletes (OA). In addition, the differences in the selfreported physician-prescribed medication use were compared between the Finnish Olympic and the Paralympic athletes. Subjects and methods: Two cross-sectional studies were conducted in Finnish Olympic athletes receiving financial support from the Finnish Olympic Committee in 2002 (n=446) and in 2009 (n=372) and in Finnish top-level Paralympic athletes (n= 92) receiving financial support from Finnish Paralympic committee in 2006. The results of the Paralympic study were compared with the results of the Olympic study conducted in 2009. Both Olympic and Paralympic athletes filled in a similar semi-structured questionnaires. Results: Dietary supplements were used by 81% of the athletes in 2002 and by 73% of the athletes in 2009. After adjusting for age-, sex- and type of sport, the odds ratio OR (95% confidence interval, CI) for use of any dietary supplement was significantly less in 2009 as compared with the 2002 situation (OR 0.62; 95% CI 0.43-0.90). Vitamin D was used by 0.7% of the athletes in year 2002 but by 2% in 2009 (ns, p = 0.07). The use of asthma medication increased from 10.4 % in 2002 to 13.7% in 2009 (adjusted OR 1.71; 95% CI 1.08-2.69). For example, fixed combinations of inhaled long-acting ��2-agonists (LABA) and inhaled corticosteroids (ICS) were used three times more commonly in 2009 than in 2002 (OR 3.38; 95% CI 1.26-9.12). The use of any physician-prescribed medicines (48.9% vs. 33.3%, adjusted OR 1.99; 95% CI 1.13-3.51), painkilling medicines (adjusted OR 2.61; 95% CI 1.18-5.78), oral antibiotics (adjusted OR 4.10; 95% CI 1.30-12.87) and anti-epileptic medicines (adjusted OR 37.09; 95% CI 5.92-232.31) was more common among the PA than in the OA during the previous seven days. Conclusions: The use of dietary supplements is on the decline among Finnish Olympic athletes. The intake of some essential micronutrients, such as vitamin D, is suprisingly low and this may even cause harm in those well-trained athletes. The use of asthma medication, especially fixed combinations of LABAs and ICS, is clearly increasing among Finnish Olympic athletes. The use of any physician-prescribed medicine, especially those to treat chronic diseases, seems to be more common among the Paralympians than in the Olympic athletes.

Novus orbis sive America meridionalis et septentrionalis

Irtokartta

Globalization of pharmaceutical R&D. Pharmaceutical R&D governance forms in People's Republic of China

Yrityksen selviytyminen ja menestyminen ovat riippuvaisia sen kyvyst�� innovoida, luoda tietoa ja hy��dynt���� tiet��myst�� ja keksint��j�� (Dunk ja Kilgore 2001). Yrityksen menestyminen erityisesti korkean teknologian alalla on siten suoraan riippuvainen sen T&T:st��, johon tehdyt investoinnit tuovat merkitt��vi�� taloudellisia etuja yritykselle uusien tuotteiden, palveluiden ja prosessien muodossa (McEvily ja Chakravarthy 1999). Teknologinen etumatka ja sen tuotteistaminen innovatiivisiksi tarjoamiksi mahdollistaa monopolististen etujen saavuttamisen yrityksen kansainv��lisess�� kilpailussa (Lall 1977). T��m�� kaltainen kilpailuetu voidaan saavuttaa yrityksen kyvyll�� yhdist���� maantieteellisesti hajautettu T&T:ns�� tehokkaaksi verkostoksi (Porter 1986). Boehen (2008) mukaan T&T:n globalisoitumista voidaan johtaa eri hallint��muodoilla: T&T:n kansainv��listymisell��, T&T:n ulkomaille sijoittamisella ja T&T ulkomaille ulkoistamisella. T&T:n globalisoituminen on osa 2000-luvun taloudellista muutosta, ja sille on esitetty useita vaikuttavia tekij��it��, kuten kustannuserot, ty��voimaresurssit, erityisosaamiskeskukset, paikallinen teknologia osaaminen ja kohdemarkkinoiden potentiaali (bardhan 2006; Norwood, ym. 2006; von Zedtwitz ja Gassmann 2002). Tutkimuksen on osoitettu eroavan tuotekehityksest�� ja eri tekij��iden on osoitettu vaikuttavan niihin (von Zedtwitz ja Gassmann 2002; Leifer ja Triscari 1987). Samoin T&T on osoitettu olevan jatkumo perustavanlaatuisesta soveltavaan ja l����kekehityksen muodostavan vastaavan T&T jatkumon (Lall 1980; Iansiti 1993), jonka yksitt��iset osat vaikuttavat sen hallintomuotoon. Tutkimus esitt���� eri tekij��it�� voivan hy��dynt���� hallintomuodosta riippuen. T��t�� tutkimusta varten tutkija haastatteli l����keteollisuuden johtajia Kiinassa vahvistaakseen tai hyl��t��kseen eri tekij��it�� ja niiden suhdetta l����keteollisen T&T:n hallintomuotoihin. Markkinoiden todettiin olevan ensisijainen tekij�� mutta my��s kustannuserojen, insentiivien, ty��voimaresurssien ja erityisosaamiskeskusten merkitys T&T:n globalisoitumiseen vaikuttavina tekij��in�� vahvistettiin yhdess�� perusvaatimusten ja riskitekij��iden kanssa. Tutkimuksessa vahvistetaan my��s l����keteollisen T&T-jatkumon vaikutus ja esitet����n viitekehys hallintomuodoille.

Globalization of pharmaceutical R&D Pharmaceutical R&D governance forms in People's Republic of China

Firm's survival and success, which are dependent on its ability to innovate, to create knowledge and to capitalize on inventions and know-how, is in essence directly linked to its R&D process (Dunk and Kilgore 2001). Especially in technology driven industries, such as the pharmaceuticals, there are signi���cant positive returns to R&D investments through introduction of new or improved products and services (McEvily and Chakravarthy 1999). Technological lead and its transformation to innovative products as fruits of corporate R&D can be seen as monopolistic advantage that helps enterprises to compete in today���s market (Lall 1977). This competitive advantage can be derived from corporation's ability to integrate its activities across geographic locations (Porter 1986). According to Boehe (2008) globalization of R&D can executed with different governance forms: R&D internationalization, R&D offshoring or R&D offshore outsourcing. Globalization of R&D is intervened with the changes in global economy of the 21st century. Some studies argue for its influencing factors to be access to vast skilled labor pools and centers of excellence (Bardhan 2006). Other studies indicate the R&D cost differentials between countries to be the major expected benefit (Norwood et al. 2006). Von Zedtwitz and Gassmann (2002) presented benefits as divided to accessing markets and customers or to accessing local science and technology. This study proposes that based on governance form distinct factor derived benefits can be capitalized. To corroborate or refute factors and their relations on R&D globalization governance forms, an empirical study based on expert interviews of pharmaceutical directors was conducted in the People's Republic of China. The market was found to be the major influencing factor. Local requirements and adaptation were corroborated as factors connected with markets. Furthermore, influencing factors, such as labour, centers of excellence, cost, financial incentives were corroborated together with conditional and risk factors. Furthermore this research argues that the globalization of pharmaceutical R&D is dependent on the financial, scientific and operational requirements of the drug discovery stage. And thus establishes the influence of drug discovery's stages continuum on pharmaceutical R&D globalization. Finally, a R&D globalization governance form decision framework is proposed based on the frameworks presented in literature and author's corroborated empirical findings.