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Introduction One third of patients with myelodysplastic syndrome (MDS) will relapse after allogeneic stem cell transplantation (SCT), with a dismal prognosis. Early detection of relapse enables pre-emptive treatment and may potentially reduce relapse risk, but is limited by the lack of sensitive markers for minimal residual disease (MRD). We developed a pipeline where patient-specific mutations, as determined by a myeloid next generation sequencing (NGS) panel are tracked using sensitive digital droplet PCR (ddPCR). Method We designed a prospective Nordic study (NMDSG14B; NCT02872662) enrolling all patients with MDS, mixed MDS / MPN or AML with myelodysplasia related disease and < 30% marrow blasts undergoing SCT in the Nordic region. We hypothesized that personalized MRD detection by ddPCR can predict clinical relapse earlier than conventional methods. Patients were included before SCT and serial bone marrow samples were collected before, and 1 and 3 months post SCT, and thereafter every third month for 2 years or until relapse or death. Blood samples were collected monthly. The MRD results were not available for the treating physicians. MRD positivity was defined based on the background noise of the specific ddPCR-assays and varied between 0.05-0.1% VAF. Results Three-hundred and sixteen patients were screened between 2016 and 2020, of which 19 were excluded due to lack of mutation or disease progression preventing SCT. We here present data of 254 patients followed ≥ 6 months after SCT. Median age was 64 (18-78) years and 59% were male. Most WHO subgroups of MDS (n=166), MDS/MPN (n=39), AML (n=8) and therapy-related disease (n=41), were represented. Risk profile according to IPSS-R was very low (n=13), low (n=32), intermediate (n=46), high (n=60) and very high (n=32). The majority of patients received pre-SCT treatment consisting of HMA (n=159) and / or intensive chemotherapy (n=59) while 60 patients did not receive disease-modifying treatment prior to SCT. The most common mutations were ASXL1 (n=69), TET2 (n=58), SRSF2 (n=57) and TP53 (n=44). No mutation was identified in 10 pts, and NGS data is still pending for 11 patients. After a median follow-up of 436 days, estimated 2 years overall survival and relapse free survival were 72% and 63%, respectively. Cumulative incidence of NRM and relapse at 2 years was 16% and 20%, respectively. Forty-six patients relapsed after a median of 170 (53-733) days, and the estimated median survival following relapse was 197 days. The most common pre-SCT mutations in the relapsed cohort were TP53 (n=19), DNMT3A (n=11) and RUNX1 (n=9). Thirty-seven patients died due to non-relapse mortality (NRM) after a median of 83 (4-754) days. To date, MRD results are available for 64 patients. Relapse was preceded by positive MRD in 14 out of 15 patients a median of 79 (21-173) days before clinical relapse. The 15th patient had an extra-medullary relapse only. Borderline positive MRD samples < 0.2% VAF within 100 days after SCT followed by negative samples were seen in 11 non-relapse patients. Twenty-four of 37 patients in continuous complete remission (CCR) were consistently MRD neg. Six CCR patients had positive MRD after 100 days; two with transient borderline peaks ( 0.1%, which turned negative when the patients developed GVHD; one patient with slowly decreasing MRD which turned negative first after 1 y, and finally one patient with prevailing KIT mutation (>700 days post-SCT) despite negative BCOR and STAG2. Two MRD+ patients died from NRM without showing signs of clinical relapse. Discussion In summary, we show that our pipeline of personalized MRD-assessment, based on patient-specific mutations is feasible with a high sensitivity to predict relapse. An update of study progression will be presented at the meeting. Figure 1 Disclosures Illman: Sanofi-Genzyme: Other: Travel Support; Celgene: Other: Travel Support. Mielke:DNA Prime: Honoraria, Other: received via my institution , Speakers Bureau; KIADIS Pharma: Honoraria, Other: received via my institution , Speakers Bureau; Miltenyi: Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: received via my institution , Speakers Bureau; Kite/Gilead: Honoraria, Other: received via my institution , Speakers Bureau; Bellicum: Honoraria, Other: received via my institution, Speakers Bureau; Novartis: Honoraria, Other: received via my institution, Speakers Bureau; Celgene/BMS: Honoraria, Other: received via my institution , Speakers Bureau. Ebeling:Accord Healthcare: Other: Travel Support; Amgen: Other: Travel Support; Celgene: Consultancy, Membership on an entity's Board of Directors or advisory committees, Other: Travel Support; Otsuka Pharma Scandanavia AB: Consultancy, Membership on an entity's Board of Directors or advisory committees.

In myelodysplastic syndrome (MDS) and myeloproliferative neoplasm (MPN), bone marrow (BM) histopathology is assessed to identify dysplastic cellular morphology, cellularity, and blast excess. Yet, other morphologic findings may elude the human eye. We used convolutional neural networks to extract morphologic features from 236 MDS, 87 MDS/MPN, and 11 control BM biopsies. These features predicted genetic and cytogenetic aberrations, prognosis, age, and gender in multivariate regression models. Highest prediction accuracy was found for TET2 [area under the receiver operating curve (AUROC) = 0.94] and spliceosome mutations (0.89) and chromosome 7 monosomy (0.89). Mutation prediction probability correlated with variant allele frequency and number of affected genes per pathway, demonstrating the algorithms' ability to identify relevant morphologic patterns. By converting regression models to texture and cellular composition, we reproduced the classical del(5q) MDS morphology consisting of hypolobulated megakaryocytes. In summary, this study highlights the potential of linking deep BM histopathology with genetics and clinical variables. Significance: Histopathology is elementary in the diagnostics of patients with MDS, but its high-dimensional data are underused. By elucidating the association of morphologic features with clinical variables and molecular genetics, this study highlights the vast potential of convolutional neural networks in understanding MDS pathology and how genetics is reflected in BM morphology. See related commentary by Elemento, p. 195.

Tiivistelmä. Tämän tutkielman tarkoitus on koota ajankohtaista tietoa yhteistoiminnallisesta oppimisesta ja siihen liittyvistä toimintatavoista, sekä siitä, miten tämä opetustapa ilmenee kouluissa. Tavoitteena on avata yhteistoiminnallisen oppimisen tausta-ajatusta vuorovaikutuksesta oppimisessa ja myös selvittää yhteistoiminnallisen oppimisen eri menetelmiä sekä työskentelytapoja. Yhteistoiminnallinen oppiminen koostuu ihmisten välisestä vuorovaikutuksesta, joten tämän käsitteen aukaiseminen on välttämätöntä, jotta yhteistoiminnallisen oppimisen idea ymmärretään oikein. Saloviita on (2006) määritellyt yhteistoiminnallisen oppimisen olevan suoraa vuorovaikusta, jossa tärkeässä roolissa on ryhmän jäsenten positiivinen riippuvuus toisistaan sekä yksilöllinen vastuu. Tämän vuorovaikutuksen aikaansaannoksena syntyy yhteinen tuotos. Yhteistoiminnallinen oppiminen on Kaganin mukaan erotettavissa muista ryhmätyöskentelymuodoista tunnuspiirteiden kautta. Näitä tunnuspiirteitä on samanaikainen vuorovaikutus, yksilöllinen vastuu, positiivinen keskinäisriippuvuus ja yhtäläinen osallistuminen (Viitattu lähteessä Kagan & Kagan, 1999). Tutkielmassa ilmeni, että yhteistoiminnallista opetusta käyttämällä opetuksesta saadaan moniulotteisempaa sekä kiinnostavampaa. Oppimisen yhteydessä oppilaat pääsevät kokemaan rakentavia keskusteluita ja kuulemaan mielenkiintoisia näkökulmia muilta oppilailta. Oppilaat pääsevät tuottamaan yhteisiä tuotoksia kantamalla yksilöllistä vastuuta mukana, joka takaa sen, että yhdessä päästään ainutlaatuisiin lopputuloksiin (Hellström, Johnson, Leppilampi & Sahlberg, 2015). Yhteistoiminnallisen oppimisen menetelmiä käyttäessä oppilaat oppivat arvokkaita taitoja, joita he tulevat tarvitsemaan tulevaisuudessa. Näitä taitoja on esimerkiksi aktiivinen osallisuus, kuunteleminen, toisten kunnioittaminen sekä itseilmaisu. Koulun arjessa yhteistoiminnallinen oppiminen ilmenee niin yksittäisinä tehtävinä kuin monen tunnin mittaisina kokonaisuuksina. Yhteistoiminnallisten menetelmien käyttöön vaikuttavat erityisesti opettajan motivaatio käyttää tätä toimintatapaa opetuksessa, sekä se, miten oppilaat käyttävät vuorovaikutustaitojaan niin, että he voisivat suoriutua yhteisistä tavoitteista.

In myelodysplastic syndrome (MDS), bone marrow (BM) histopathology is visually assessed to identify dysplastic cellular morphology, cellularity, and blast excess. Yet, many morphological findings elude the human eye. Here, we extracted visual features of 236 MDS, 87 MDS/MPN, and 10 control BM biopsies with convolutional neural networks. Unsupervised analysis distinguished underlying correlations between tissue composition, leukocyte metrics, and clinical characteristics. We applied morphological features in elastic net-regularized regression models to predict genetic and cytogenetic aberrations, prognosis, and clinical variables. By parallelizing tile, pixel, and leukocyte-level image analysis, we deconvoluted each model to texture and cellular composition to dissect their pathobiological context. Model-based mutation predictions correlated with variant allele frequency and number of affected genes per pathway, demonstrating the models’ ability to identify relevant visual patterns. In summary, this study highlights the potential of deep histopathology in hematology by unveiling the fundamental association of BM morphology with genetic and clinical determinants.

Tilaseurannassa selvitettiin valkuaislähteiden soveltuvuutta välikasvatusporsaiden liemiruokintaan, niiden vaikutuksia porsaiden tuotanto-ominaisuuksiin (päiväkasvu, rehun syönti ja rehuhyötysuhde) ja hyvinvointiin (hännänpurenta ja kuolleisuus). Tutkitut kotimaiset valkuaisrehukomponentit olivat ohravalkuaisrehu (OVR) ja härkäpapu. Seurannassa oli yhteensä 560 Duroc-kolmiroturisteytysporsasta, joiden alkupaino oli keskimäärin 9.1 kg. Porsaat jaettiin kontrolli- ja koeryhmään. Havaintoyksikkönä kokeessa oli venttiili. Jokaisella venttiilillä oli 40 porsasta. Kontrollirehu sisälsi ohraa, vehnää, kauraa, kasviöljyä, soijapohjaista täydennysrehua ja ohravalkuaisrehua. Koerehussa soijan sijaan käytettiin härkäpapua sekä härkäpapuruokinnan koostumusta täydentävää rehua. Porsailla oli kolmivaiheinen vapaa liemiruokinta ja rehunkulutusta seurattiin tilan ruokintalaitteella. Porsaat punnittiin kokeen alussa, ruokintajaksojen vaihtuessa ja kokeen lopussa. Porsaat kasvoivat keskimäärin 480 g päivässä eikä päiväkasvussa ollut tilastollisesti merkitseviä eroja ryhmien välillä. Rehunkulutus koko kokeen aikana eläintä kohti oli 342 MJ NEk ja 30.4 kg ka, eikä ryhmien välillä ollut merkitseviä eroja. Myöskään rehuhyötysuhteissa ei ryhmien välillä tullut merkitseviä eroja. Välikasvattamossa olennaista on, että porsaat kasvavat nopeasti, tasaisesti ja rehutehokkaasti välityspainoonsa. Molemmissa ruokintaryhmissä yli 75% porsaista myytiin eteenpäin 49 vuorokauden sisällä vieroituksesta. Kontrolliryhmästä suurin osa porsaista myytiin 46‒47 päivän kuluttua (54%) ja koeryhmästä 39‒40 päivän kuluttua vieroituksesta (47%). Koeryhmän porsaita jäi kuitenkin koeseurannan puitteissa merkittävästi enemmän jäljelle (21 vs 12%) suhteessa myytyjen porsaiden määrään. Tilaseurannan tulokset viittaavat siihen, että härkäpavulla ja siihen sopivilla täydennyksillä voitaisiin korvata rehun soija vilja-OVR‒pohjaisella ruokinnalla. Kontrolli- ja koeryhmän porsaiden välillä ei ollut merkittävää eroa tuotantotuloksissa. Tutkimustietoon perustuvien johtopäätösten saamiseksi vaaditaan kuitenkin jatkotutkimuksia suuremmalla havaintomäärällä tai tarkentamalla koeasetelmaa esimerkiksi porsaskohtaisin lisäpunnituksin.

ContextMutations to isocitrate dehydrogenase (IDH) appear to play a prognostic or predictive role in several neoplasias. Immunohistochemical staining designed to detect a specific R132H mutation to IDH1 showed expression in the normal adrenal cortex, raising interest to study the potential role of IDH1 in the pathogenesis of adrenocortical tumors.ObjectiveThe objective of this work is to study the role of IDH1 and its mutations in adrenocortical tumors.Design and patientsIDH1 R132H immunohistological staining was performed on a cohort of 197 adrenocortical tumors. The exon of the IDH1 gene was sequenced in 16 tumors.ResultsPositive IDH1 R132H immunohistochemical staining correlated with a better prognosis among patients with a malignant adrenocortical tumor. However, IDH1 R132H immunohistochemistry did not distinguish between local and metastasized tumors. We were unable to identify IDH1 mutations among our adrenocortical tumors using a targeted next-generation sequencing panel or via exon sequencing.ConclusionsAmong adrenocortical carcinomas, IDH1 R132H immunopositivity correlated with a better prognosis. Thus, IDH1 R132H immunohistochemical staining could serve as a prognostic or as a potential predictive marker in adrenocortical carcinomas. Further research is needed to identify the possible alterations in IDH1 that could explain our findings, because we identified no known mutations to the IDH1 gene.

This is the first in a series of articles about recovering the full algebraic structure of a boundary conformal field theory (CFT) from the scaling limit of the critical Ising model in slit-strip geometry. Here, we introduce spaces of holomorphic functions in continuum domains as well as corresponding spaces of discrete holomorphic functions in lattice domains. We find distinguished sets of functions characterized by their singular behavior in the three infinite directions in the slit-strip domains. We prove convergence results of the distinguished discrete holomorphic functions to the continuum ones. In the subsequent articles, the discrete holomorphic functions will be used for the calculation of the Ising model fusion coefficients (as well as for the diagonalization of the Ising transfer matrix), and the convergence of the functions is used to prove the convergence of the fusion coefficients. It will also be shown that the vertex operator algebra of the boundary conformal field theory can be recovered from the limit of the fusion coefficients via geometric transformations involving the distinguished continuum functions., Comment: v3: 50 pages, minor modifications and corrections

Ports have played an important role in facilitating exchanges among countries since the day when inland transportation was poor. As ports become hubs for global supply chain, they have to maintain their competitiveness not only by reassuring their efficiency, reliability, accessibility to hinterland, and sustainability. In addition, there is a constant challenge from all operational parties of the port to acquire needed information or to trust information received, due to multiple legacy systems and platforms that do not integrate with each other, and to the lack of real time updates. There are differing agendas between parties and, sometimes, distrust within the multi-stakeholder ecosystem leads to working in silos. This jeopardises seamless data exchange and cooperation across the port value chain, resulting in significant inefficiencies. Port community system (PCS) can enhance communication and simplify administrative process resulting economic and environmental benefit for actors in the supply chain. The invisibility of the benefit, actors’ heterogeneity and significant investment to develop the system resulting a reluctance in implementing PCS. This chapter aims to study the evolution mechanism behind the process of PCS network development using lessons learned from industrial symbiosis network development and network trajectories theory. The PCS network development follows a serendipitous and goal-oriented process that can be categorised into three stages: pre-PCS network, PCS network emergence, and PCS network expansion. This chapter contributes to the exploration of network evolution and documents lesson learned to foster PCS implementation.

Vertaisarvioitu. Teema : aivokasvaimet. English summary. Glioomat ovat primaareista aivokasvaimista yleisimpiä. Glioomien diagnostiikka perustuu kasvainten histologisiin piirteisiin ja molekulaarisiin muutoksiin. Vuonna 2016 päivitetty WHO:n aivokasvainluokitus tuo geneettiset muutokset osaksi kasvainten diagnoosia. Integroitu diagnoosi sisältää kasvaimen histologisen tyypin, pahanlaatuisuusasteen ja molekulaariset muutokset. Glioomien tärkeimmät molekyylimuutokset ovat IDH (isositraattidehydrogenaasi) -geenin mutaatio ja 1p/19q-kodeleetio. Astrosytoomissa IDH-mutaatio erottaa suotuisan ennusteen kasvaimet omaksi ryhmäkseen. IDH-mutatoitumaton glioblastooma on glioomista tavallisin ja pahanlaatuisin. Paremman ennusteen oligodendroglioomassa on samanaikaisesti IDH-mutaatio ja 1p/19q-kodeleetio. H3 K27M -mutatoitunut keskiviivan gliooma on uusi aggressiivinen glioomatyyppi. Molekyylimuutoksiin perustuva luokittelu parantaa glioomien diagnostiikan tarkkuutta ja luo pohjan niiden tulokselliselle hoidolle.

Circulating tumor DNA (ctDNA) is released from cancer cells and oncogenic mutations in ctDNA can be measured from plasma samples. Droplet digital PCR (ddPCR) is a sensitive and specific method for the detection of mutations in ctDNA. We analyzed serial plasma samples (n = 80) from ten metastatic colorectal cancer (mCRC) patients with a known KRAS mutation in their primary tumor. The patients were undergoing oncological treatment with bevacizumab in combination with alternating capecitabine and oxaliplatin or irinotecan. Baseline ddPCR KRAS mutation allele frequency (MAF) values ranged from 0% to 63%. The first radiologic response evaluation criteria in solid tumors (RECIST) evaluation was performed 45–63 days after the initiation of treatment, and by this time three patients had an undetectable level of KRAS mutation, one had a MAF value of 0.5%, and one had a MAF value of 3% that had been reduced by 95% from the baseline value. In three of these patients the RECIST assessment was stable disease and in two partial response. In seven patients, ddPCR MAF values increased before radiological disease progression or death, while one patient remained disease-free with an undetectable KRAS mutation level. Next, we analyzed all available plasma samples with the Idylla ctKRAS system (n = 60), and found that the overall degree of agreement between ddPCR and Idylla was almost perfect (kappa value = 0.860). We used next-generation sequencing (NGS) to detect treatment-induced mutations in the last serial plasma sample of each patient, but were unable to find any new mutations when compared to the primary tumor. This study shows that ddPCR and Idylla are equally efficient for the detection of KRAS mutations in the liquid biopsies from mCRC patients and that ctDNA may indicate the disappearance of treatment responsive KRAS positive mCRC clones and serve as an early sign of disease progression.

Tässä tutkimuksessa tarkasteltiin metropolialueen 9. luokkalaisten hyvinvointia ja koulumenestystä. On nostettu esiin huoli siitä, onko suomalaisoppilaiden hyvä menestys kansainvälisissä oppimistulostesteissä saavutettu heikentyneen hyvinvoinnin kustannuksella. Monien yksilöllisten tekijöiden on havaittu vaikuttavan koulussa menestymiseen, mutta ajankohtaista tietoa etenkin hyvinvoinnin ja terveyden vaikutuksesta tarvitaan lisää. Tässä tutkimuksessa tavoitteena oli selvittää, miten itse arvioitu terveys, stressioireet ja koulu-uupumuksen ulottuvuudet ovat yhteydessä koulumenestykseen. Tutkimuksessa hyödynnettiin metropolialueen 9. luokkalaisten hyvinvoinnin ja oppimista tarkastelleen tutkimuksen aineistoa (n=7738). Tätä pro gradu -tutkielmaa varten aineistosta kerättiin tiedot oppilaiden äidinkielen, matematiikan ja luonnontieteiden todistusarvosanoista, joista laskettua keskiarvoa käytettiin koulumenestyksen mittarina. Tutkimuksessa hyödynnettävät tiedot hyvinvoinnin ja terveyden osalta kerättiin sähköisen hyvinvointikyselyn aineistosta. Tutkimus toteutettiin määrällisenä tutkimuksena ja tutkimusmenetelminä käytettiin faktorianalyysia, varianssianalyysia sekä regressioanalyysia. Tutkimuksessa itse arvioidun terveyden ja koulumenestyksen havaittiin olevan yhteydessä toisiinsa. Itse arvioitu terveys luokiteltiin huonon, keskinkertaisen ja hyvän terveyden ryhmiin. Näiden terveysryhmien havaittiin eroavan toisistaan koulumenestyksen suhteen tilastollisesti merkitsevästi. Myös stressioireiden havaittiin selittävän koulumenestyksen vaihtelua ja tilastollisesti merkitsevä yhteys koulumenestykseen oli kuudella tutkimuksessa kartoitetulla stressin oireella. Yksittäisten stressioireiden vaikutus keskiarvon vaihteluun oli kuitenkin melko pientä. Myös koulu-uupumuksen ja koulumenestyksen välillä havaittiin olevan yhteys. Tutkimuksessa koulu-uupumuksen ulottuvuuksia kuvaavat koulun merkityksettömyys ja kuormittavuus yhdessä selittivät koulumenestyksen vaihtelusta 18 prosenttia. Tämän tutkimuksen tulosten mukaan koulumenestyksen parantuessa koulu-uupumus vähenee. Tämä tutkimus tuottaa arvokasta tietoa terveyden ja hyvinvoinnin yhteydestä koulumenestykseen. Tutkimuksen tulosten valossa koulussa menestymistä voitaisiin edesauttaa panostamalla oppilaiden terveyteen ja kokonaisvaltaiseen hyvinvointiin, johon vaikuttavat myös stressi ja koulu-uupumus. Kiinnittämällä huomiota koulutyön kuormittavuuteen ja ennaltaehkäisemällä koulusta johtuvaa stressiä ja uupumusta voidaan vaikuttaa koulumenestykseen, jolla on suuri merkitys paitsi yksilön elämänkulun kannalta, myös yhteiskunnallisella tasolla.

The purpose of this thesis is to highlight why construction materials should be reclaimed and how to reuse them in future architectural projects, drawing ideas from examples in fashion and existing buildings. The relationship between architecture and fashion was studied to explore their close connection and that ideas can be transferred from one field to another. The current practices of the building industry are unsustainable and wasteful: a building is demolished to possibly build a new one on its place and the materials of the old building are thrown away, even if they are not at the end of their useful age yet. We should change this way of thinking and the built-up value and the aesthetics of used materials should be recognized and appreciated. Ideas of reclamation were found from examples in existing buildings and fashion through a case study analysis. They were distilled into categories based on the reclamation techniques to clarify the differences between different methods. Many lessons can be learned from the reclamation architecture and fashion examples to apply to the practices in the field of architecture. There are still some practical obstacles to overcome, when it comes to making the practice widespread and mainstream. These include scaling up the practice of reclamation and managing the supply and demand, as well as everyone in the building industry taking equal responsibility to be transparent about their actions and efforts at sustainability and reclamation. Tämän diplomityön tarkoitus on valottaa syitä rakennusmateriaalien uudelleenkäyttöön, ja tutkia muoti- ja arkkitehtuuriesimerkkien perusteella, miten niitä voi hyödyntää luovasti tulevaisuuden rakennushankkeissa. Muodin ja arkkitehtuurin välistä suhdetta tarkasteltiin uusien ideoiden löytämiseksi alalta toiselle niiden läheisen yhteyden vuoksi. Rakennusalan nykykäytännöt ovat kestämättömiä ja tuhlailevia: rakennuksia puretaan uusien tieltä ja vanhojen rakennuksen materiaaleja ei hyödynnetä, vaikka ne eivät ole vielä käyttöikänsä päässä. Tällaisen ajattelutavan on muututtava. Meidän tulisi arvostaa materiaaleja, niiden estetiikkaa ja niihin kertynyttä arvoa, sekä kunnioittaa niiden jalostamiseen käytettyjä raaka-aineita ja vaivannäköä. Esimerkillisiä ideoita uudelleenkäytöstä etsittiin tapaustutkimuksella olemassa olevista rakennuksista ja muodista. Löydettyjen piirteiden perusteella eri uudelleenkäyttötekniikat jaettiin kategorioihin erojen havainnollistamiseksi ja tutkimiseksi. Näistä tekniikoista, niin uudelleenkäyttöarkkitehtuurissa, kuin uudelleenkäyttömuodissakin, voidaan oppia paljon sovellettavaksi arkkitehtuuriin tulevaisuudessa. Uudelleenkäyttö ei ole vielä valtavirtaa rakennusalalla. Tämä edellyttää uudelleenkäytön mittakaavan kasvattamista suuremmaksi, kysynnän ja tarjonnan kohtaamista, sekä tasapuolista vastuunottoa ja läpinäkyvyyttä uudelleenkäytön ja kestävyyden tavoittelussa rakennusalalla.

Traditionally grass has been considered a suitable feed only for ruminants. Due to global challenges related to growing human population, sustainable feed materials for pigs are actively searched and grasses as a high yielding biomass have been seen as an interesting addition to pig feeds. The digestible nutrients in grasses are embedded in a fibre matrix not digestible to pigs, but suitable processing could alleviate this. The aim of the present study was to evaluate whether silage juice could provide a new sustainable diet ingredient for fattening pigs. Grass silage was fractionated with a twin-screw press to liquid and solid fractions. The silage juice was included into liquid feed of 112 fattening pigs from 62 kg live weight until slaughter (124 kg live weight) and compared to 96 pigs on a conventional control diet. Feed allowance was ad libitum until it reached 33.6 MJ net energy day−1 per pig which was kept as the maximum feed allowance until slaughter. The average daily gain of all the pigs was over 1000 g day−1 in the experimental period but no difference between silage juice fed and control group was found. The feed conversion ratio was, as expected, higher in barrows than gilts, but no difference was found between the two treatment groups. Variation in silage juice dry matter content during the experimental period may have led to a lower dry matter intake of silage juice fed pigs. Gilts had statistically significantly higher meat percentage than barrows in both treatments, but no difference was found between the treatment groups. Meat quality (pH, colour, drip loss and sensory analyses) results were similar in both treatment groups. For gut microbiota, no statistically significant differences were found in any of the bacterial families studied (Actinobacteriota, Bacteroidota, Firmicutes, Deferrivacterota, Spirochaetota, Minor Phyla). In conclusion, silage juice is a potential feed ingredient in liquid feed for pigs. Microbiological data indicates that silage juice does not cause gastrointestinal microbiota imbalance in pigs.

Background BCL-2 inhibitor venetoclax (Ven) in combination with azacitidine (Aza) has substantially improved overall response rates (ORR) and overall survival (OS) in newly diagnosed nonfit acute myeloid leukemia (AML). Nevertheless, 1/3 of patients (pts) do not respond to the treatment, and for relapsed/refractory (R/R) pts the ORR is 20‒50%. While certain genetic changes correlate with treatment outcome (e.g., IDH2, TP53), mutations do not precisely predict Ven-Aza response. Thus, novel approaches are needed to identify those who benefit from Ven-Aza, particularly in the R/R setting. Ex vivo drug sensitivity has been explored in small pilot studies to guide treatment in R/R AML. However, very few prospective clinical trials have analyzed the predictability of ex vivo drug sensitivity testing. It is also unknown whether different drug testing platforms (e.g., flow cytometry (FC) and luminescent cell viability assays) and culture conditions influence the predictive accuracy. Therefore, we designed a prospective trial to evaluate the ex vivo/in vivo correlation of drug sensitivity testing with the Ven-Aza treatment outcome in AML. Methods VenEx is an ongoing, multicenter, open-label phase II study by the Finnish AML Group (NCT04267081) that includes nonfit de novo, R/R, and secondary AML (sAML) pts. Here, we report the interim results from the first part of the trial (validation cohort), in which all patients received study treatment regardless of the ex vivo drug sensitivity testing results (n = 39). All pts received Aza (75 mg/m 2 on days 1‒7, 28-day cycle) and Ven (400 mg once daily, 28-day cycles, shortened in responding pts to 21 days). The primary endpoint was cumulative ORR (CR/CRi/MLFS) during the first three treatment cycles, and a key secondary endpoint was the correlation of ex vivo drug sensitivity with ORR and OS. For drug sensitivity testing, 20 ml of bone marrow (BM) was shipped to the central laboratory, and Ficoll-purified mononuclear cells (MNCs) were plated within 26 hours after sampling. The drug plates contained Ven, Ven + Aza, and other BCL-2 family inhibitors across a 10,000-fold concentration range. Ven sensitivity was measured in three cell culture conditions: 1. RPMI, 2. conditioned medium (CM, HS-5 cell line derived medium (Karjalainen et al., 2017), and 3. StemSpan SFEMII + 20 ng/ml of FLT3L + SCF + TPO (SPM). After 48 h of incubation with the drugs, cell viability was measured in parallel with CellTiter-Glo (CTG) and FC, as described by Kuusanmäki et al. (2020). Drug sensitivity score (DSS) derived from the area under the dose response curve calculations indicated efficacy (Yadav et al., 2014). Results In the validation cohort, 16 pts had de novo AML and 23 had sAML or R/R AML. The ORR for de novo AML was 88% (14/16) and for R/R or sAML 52% (12/23). Drug sensitivity testing was successful in 38/39 pts. The best correlation of responding and non-responding pts occurred when blast-specific Ven sensitivity was measured by FC in CM media (p < 0.001, Figures A-B). When the ex vivo/in vivo correlation with CTG (BM bulk MNC sensitivity) was assessed, several pts who had achieved CR/CRi/MLFS during the first three cycles were resistant to Ven ex vivo (Figure A). Similarly, the SPM medium yielded false negative ex vivo responses in blast-specific FC measurement. Ex vivo testing of the Ven-Aza combination by FC did not increase the predictive accuracy over Ven alone (Figure A). Using a DSS threshold of 10.7 for blast-specific FC Ven sensitivity in CM, the test's sensitivity was 92%, its specificity was 75%, and its positive predictive value was 88% (Figure C). The median OS for patients who had DSS < 10.7 (ex vivo resistant) was 3.5 months, and those with DSS > 10.7 (ex vivo sensitive) have not reached the median OS (p < 0.001, Figure D). Conclusions Ex vivo drug sensitivity testing was feasible for AML pts in a clinical trial context. Leukemic blast-specific ex vivo Ven sensitivity showed a high correlation with ORR, with a positive predictive value of 88%, and predicted longer OS. The interim results argue that ex vivo Ven sensitivity can be used as a predictive biomarker for Ven-Aza therapy. Notably, experimental conditions had considerable influence on the predictive value of ex vivo Ven sensitivity; thus, between-trial standardization is crucial. The ongoing VenEx trial will validate these findings in the second part of the trial (60 pts) where ex vivo Ven sensitivity guides therapy in pts with R/R or sAML. Figure 1 Figure 1. Disclosures Kuusanmäki: AbbVie: Research Funding. Heckman: Novartis: Research Funding; Orion Pharma: Research Funding; Celgene/BMS: Research Funding; Oncopeptides: Consultancy, Research Funding; Kronos Bio, Inc.: Research Funding. Ettala: Novartis: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees; Amgen: Other: Conference expensens; Sanofi: Other: Conference expensens. Pyörälä: Pfizer: Honoraria; Novartis: Honoraria; AbbVie: Honoraria. Rimpiläinen: Astellas: Membership on an entity's Board of Directors or advisory committees; AbbVie: Membership on an entity's Board of Directors or advisory committees; Pfizer: Membership on an entity's Board of Directors or advisory committees. Siitonen: amgen: Honoraria; Novartis: Honoraria, Membership on an entity's Board of Directors or advisory committees; Brystol Myers Squibb: Consultancy; Janssen-Cilag: Consultancy, Membership on an entity's Board of Directors or advisory committees; abbvie: Membership on an entity's Board of Directors or advisory committees; Pfizer: Consultancy, Membership on an entity's Board of Directors or advisory committees; celgene: Membership on an entity's Board of Directors or advisory committees. Kontro: Astellas: Consultancy, Membership on an entity's Board of Directors or advisory committees; Jazz Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees; AbbVie: Membership on an entity's Board of Directors or advisory committees, Research Funding.

Fast progress of the next generation sequencing (NGS) technology has allowed global transcriptional profiling and genome-wide mapping of transcription factor binding sites in various cellular contexts. However, limited number of replicates and high amount of data processing may weaken the significance of the findings. Comparative analyses of independent data sets acquired in the different laboratories would greatly increase the validity of the data. Runx2 is the key transcription factor regulating osteoblast differentiation and bone formation. We performed a comparative analysis of three published Runx2 data sets of chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) analysis in osteoblasts from mouse and human origin. Moreover, we assessed the similarity of the corresponding transcription data of these studies available online. The ChIP-seq data analysis confirmed general features of Runx2 binding, including location at genic vs intergenic regions and abundant Runx2 binding on promoters of the highly expressed genes. We also found high frequency of Runx2 DNA binding without a consensus Runx2 motif at the binding site. Importantly, mouse and human Runx2 showed moderately similar binding patterns in terms of peak-associated closest genes and their associated genomic ontology (GO) pathways. Accordingly, the gene expression profiles were highly similar and osteoblastic phenotype was prominent in the differentiated stage in both species. In conclusion, ChIP-seq method shows good reproducibility in the context of mature osteoblasts, and mouse and human osteoblast models resemble each other closely in Runx2 binding and in gene expression profiles, supporting the use of these models as adequate tools in studying osteoblast differentiation.

In many cancer types, integrin-mediated signaling regulates proliferation, survival and invasion of tumorigenic cells. However, it is still unclear how integrins crosstalk with oncogenes to regulate tumorigenesis and metastasis. Here we show that oncogenic K-RasV12 upregulates α6-integrin expression in Madin–Darby canine kidney (MDCK) cells via activation of the mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK)/Fos-related antigen 1-signaling cascade. Activated α6-integrins promoted metastatic capacity and anoikis resistance, and led to perturbed growth of MDCK cysts. Transcriptomic analysis of K-RasV12-transformed MDCK cells also revealed robust downregulation of αV-class integrins. Re-expression of αV-integrin in K-RasV12-transformed MDCK cells synergistically upregulated the expression of Zinc finger E-box-binding homeobox 1 and Twist-related protein 1 and triggered epithelial-mesenchymal transition leading to induced cell motility and invasion. These results delineate the signaling cascades connecting oncogenic K-RasV12 with α6- and αV-integrin functions to modulate cancer cell survival and tumorigenesis, and reveal new possible strategies to target highly oncogenic K-RasV12 mutants.

Background Solid tumors residing in tissues and organs leave footprints in circulation through circulating tumor cells (CTCs) and circulating tumor DNAs (ctDNA). Characterization of the ctDNA portraits and comparison with tumor DNA mutational portraits may reveal clinically actionable information on solid tumors that is traditionally achieved through more invasive approaches. Methods We isolated ctDNAs from plasma of patients of 103 lung cancer and 74 other solid tumors of different tissue origins. Deep sequencing using the Guardant360 test was performed to identify mutations in 73 clinically actionable genes, and the results were associated with clinical characteristics of the patient. The mutation profiles of 37 lung cancer cases with paired ctDNA and tumor genomic DNA sequencing were used to evaluate clonal representation of tumor in circulation. Five lung cancer cases with longitudinal ctDNA sampling were monitored for cancer progression or response to treatments. Results Mutations in TP53, EGFR, and KRAS genes are most prevalent in our cohort. Mutation rates of ctDNA are similar in early (I and II) and late stage (III and IV) cancers. Mutation in DNA repair genes BRCA1, BRCA2, and ATM are found in 18.1% (32/177) of cases. Patients with higher mutation rates had significantly higher mortality rates. Lung cancer of never smokers exhibited significantly higher ctDNA mutation rates as well as higher EGFR and ERBB2 mutations than ever smokers. Comparative analysis of ctDNA and tumor DNA mutation data from the same patients showed that key driver mutations could be detected in plasma even when they were present at a minor clonal population in the tumor. Mutations of key genes found in the tumor tissue could remain in circulation even after frontline radiotherapy and chemotherapy suggesting these mutations represented resistance mechanisms. Longitudinal sampling of five lung cancer cases showed distinct changes in ctDNA mutation portraits that are consistent with cancer progression or response to EGFR drug treatment. Conclusions This study demonstrates that ctDNA mutation rates in the key tumor-associated genes are clinical parameters relevant to smoking status and mortality. Mutations in ctDNA may serve as an early detection tool for cancer. This study quantitatively confirms the hypothesis that ctDNAs in circulation is the result of dissemination of aggressive tumor clones and survival of resistant clones. This study supports the use of ctDNA profiling as a less-invasive approach to monitor cancer progression and selection of appropriate drugs during cancer evolution. Electronic supplementary material The online version of this article (doi:10.1186/s13045-017-0468-1) contains supplementary material, which is available to authorized users.

The focus in this volume is on social media discourse, (dis)identifications and diversities. It demonstrates how particular ways of mobilizing verbal, discursive and other semiotic resources serve as means for identity work (Bucholtz, 2003; Blommaert, 2003), involving acts, processes and practices of (dis)identification as essential aspects of sociality in social media. It will also illustrate how such social action also increasingly engages with a range of diversities in social media. peerReviewed

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Anti-EGFR antibodies are used for the treatment of RAS wild type metastatic colorectal cancer. We previously showed that EGFR gene copy number (GCN) predicts response to anti-EGFR therapy in KRAS exon 2 wild type metastatic colorectal cancer. The aim of our study was to analyse the predictive role of EGFR GCN in RAS/BRAF/PIK3CA wild type metastatic colorectal cancer. The material included 102 patients with KRAS exon 2 wild type metastatic colorectal cancer treated with anti-EGFR ± cytotoxic therapy. Next generation sequencing was used for KRAS, NRAS, BRAF and PIK3CA gene mutation analyses. EGFR GCN was analysed by EGFR immunohistochemistry guided automated silver in situ hybridisation. Increased EGFR GCN (≥4.0) predicted a better response and prolonged progression free survival in anti-EGFR treated RAS/BRAF/PIK3CA wild type patients (Log-rank test, p = 0.0004). In contrast, survival of RAS/BRAF/PIK3CA wild type, EGFR GCN below 4.0 patients did not differ from patients with mutant RAS, BRAF or PIK3CA. Our study indicates that EGFR GCN predicts anti-EGFR treatment efficacy in patients with RAS/BRAF/PIK3CA wt metastatic CRC. Tumours with EGFR GCN below 4.0 appear to be as refractory to anti-EGFR treatment as tumours with mutation in any of the RAS/RAF/PIK3CA pathway genes.

Strategic thinking is considered a significant business process by management experts due to it's appeal to strengthen organizational performance management and its effectiveness. Furthermore, new product development is regarded as the crucial operation of each and every industrial concern. Grounded in such rationality, the current study presents an attempt to connect the concept of strategic thinking with product innovativeness to formulate a diagnostic approach to reinforce NPD support process. The idea behind formulating such a research framework is to offer industrial leaders, a mechanism, that can offer connectivity among various aspects of operations linked to corporate strategy and product innovativeness. The research survey is conducted on 30 team member's representing new product development (NPD) operations at three selected work locations of a European multinational company. The study results highlighted the proposed theoretical connection between the two business operational areas; strategic thinking and product innovation. According to the study results, the key variables that have shown positive linkage among them were, early client involvement (product innovation) to target reach (strategic thinking); customer value (product innovation) to target reach (strategic thinking); early client involvement (product innovation) to customer value (Product innovation); management initiatives (product innovation) to collaboration (strategic thinking); management initiatives (product innovation) to communication (strategic thinking); and idea generation (product innovation) to responsiveness (strategic thinking).

The proprotein convertase enzyme FURIN processes immature pro-proteins into functional end- products. FURIN is upregulated in activated immune cells and it regulates T-cell dependent peripheral tolerance and the Th1/Th2 balance. FURIN also promotes the infectivity of pathogens by activating bacterial toxins and by processing viral proteins. Here, we evaluated the role of FURIN in LysM+ myeloid cells in vivo. Mice with a conditional deletion of FURIN in their myeloid cells (LysMCre-fur(fl/fl)) were healthy and showed unchanged proportions of neutrophils and macrophages. Instead, LysMCre-fur(fl/fl) mice had elevated serum IL-1β levels and reduced numbers of splenocytes. An LPS injection resulted in accelerated mortality, elevated serum pro-inflammatory cytokines and upregulated numbers of pro-inflammatory macrophages. A genome-wide gene expression analysis revealed the overexpression of several pro-inflammatory genes in resting FURIN-deficient macrophages. Moreover, FURIN inhibited Nos2 and promoted the expression of Arg1, which implies that FURIN regulates the M1/M2-type macrophage balance. FURIN was required for the normal production of the bioactive TGF-β1 cytokine, but it inhibited the maturation of the inflammation-provoking TACE and Caspase-1 enzymes. In conclusion, FURIN has an anti-inflammatory function in LysM+ myeloid cells in vivo.