Screening for potential undiagnosed Gaucher disease patients : Utilisation of the Gaucher earlier diagnosis consensus point-scoring system (GED-C PSS) in conjunction with electronic health record data, tissue specimens, and small nucleotide polymorphism (SNP) genotype data available in Finnish biobanks

Background: Autosomal recessive Gaucher disease (GD) is likely underdiagnosed in many countries. Because the number of diagnosed GD patients in Finland is relatively low, and the true prevalence is currently not known, it was hypothesized that undiagnosed GD patients may exist in Finland. Our previous study demonstrated the applicability of Gaucher Earlier Diagnosis Consensus point-scoring system (GED-C PSS; Mehta et al., 2019) and Finnish biobank data and specimens in the automated point scoring of large populations. An indicative point -score range for Finnish GD patients was determined, but undiagnosed patients were not identified partly due to high number of high-score subjects in combination with a lack of suitable samples for diagnostics in the assessed biobank population. The current study extended the screening to another biobank and evaluated the feasibility of utilising the automated GED-C PSS in conjunction with small nucleotide polymorphism (SNP) chip genotype data from the FinnGen study of biobank sample donors in the identification of undiagnosed GD patients in Finland. Furthermore, the applicability of FFPE tissues and DNA restoration in the next-generation sequencing (NGS) of the GBA gene were tested. Methods: Previously diagnosed Finnish GD patients eligible to the study, and up to 45,100 sample donors in Helsinki Biobank (HBB) were point scored. The GED-C point scoring, adjusted to local data, was automated, but also partly manually verified for GD patients. The SNP chip genotype data for rare GBA variants was visually assessed. FFPE tissues of GD patients were obtained from HBB and Biobank Borealis of Northern Finland (BB). Results: Three previously diagnosed GD patients and one patient previously treated for GD-related features were included. A genetic diagnosis was confirmed for the patient treated for GD-related features. The GED-C point score of the GD patients was 12.5-22.5 in the current study. The score in eight Finnish GD patients of the previous and the current study is thus 6-22.5 points per patient. In the automated point scoring of the HBBPeer reviewe

Real-time detection of cerebrospinal fluid with bioimpedance needle in paediatric lumbar puncture

Background Lumbar puncture is a common clinical procedure that can occasionally be difficult. Various needle guidance methods can facilitate performing this procedure, but at the expense of special expertise, equipment and facility. In the present study, we evaluated the clinical feasibility of a novel bioimpedance needle system regarding its ability to detect cerebrospinal fluid (CSF) in paediatric lumbar punctures.Methods We performed 40 lumbar puncture procedures using the bioimpedance needle system in 37 paediatric patients, aged from 0 days to 17 months, as a part of their prescribed examinations in two university hospitals. The bioimpedance needle is similar to a conventional 22G cutting-edge spinal needle with a stylet, except the needle and stylet are configured as a bipolar electrode with high spatial resolution. The system measures in real-time when the needle tip reaches the subarachnoid space containing CSF. The procedure was considered successful when the erythrocyte count was determined from the obtained CSF sample.Results Subarachnoid space was verifiably reached in 28 out of 40 procedures (70%). Bioimpedance needle system detected CSF in 23 out of these 28 successful procedures (82%) while failed in 3 out of 28 procedures (11%). No adverse events were reported.Conclusion Bioimpedance spinal needle system was found clinically feasible in paediatric lumbar punctures, and it may offer an objective and simple means to detect the time point when the needle tip is in contact with the cerebrospinal fluid.</div

Prognostic significance of spatial and density analysis of T lymphocytes in colorectal cancer

Background Although high T cell density is a strong favourable prognostic factor in colorectal cancer, the significance of the spatial distribution of T cells is incompletely understood. We aimed to evaluate the prognostic significance of tumour cell-T cell co-localisation and T cell densities. Methods We analysed CD3 and CD8 immunohistochemistry in a study cohort of 983 colorectal cancer patients and a validation cohort (N = 246). Individual immune and tumour cells were identified to calculate T cell densities (to derive T cell density score) and G-cross function values, estimating the likelihood of tumour cells being co-located with T cells within 20 mu m radius (to derive T cell proximity score). Results High T cell proximity score associated with longer cancer-specific survival in both the study cohort [adjusted HR for high (vs. low) 0.33, 95% CI 0.20-0.52, P-trend < 0.0001] and the validation cohort [adjusted HR for high (vs. low) 0.15, 95% CI 0.05-0.45, P-trend < 0.0001] and its prognostic value was independent of T cell density score. Conclusions The spatial point pattern analysis of tumour cell-T cell co-localisation could provide detailed information on colorectal cancer prognosis, supporting the value of spatial measurement of T cell infiltrates as a novel, robust tumour-immune biomarker.Peer reviewe

CD3+ and CD8+ T-Cell-Based Immune Cell Score and PD-(L)1 Expression in Pulmonary Metastases of Microsatellite Stable Colorectal Cancer

The objective of this study was to evaluate the prognostic value of CD3+ and CD8+ based immune cell score (ICS), programmed death -1 (PD-1) and programmed death ligand -1 (PD-L1) in pulmonary metastases of proficient mismatch repair colorectal cancer (CRC) patients. A total of 101 pulmonary metastases and 62 primary CRC tumours were stained for CD3+, CD8+, PD-1 and PD-L1 expression. The prognostic value of ICS, PD-1/PD-L1 expression in 67 first pulmonary metastases and 61 primary CRC tumour was analysed. Comparative analysis was also performed between primary tumours and pulmonary metastases, as well as between T-cell densities and PD-1/PD-L1 expression. The 5-year overall survival rates of low, intermediate, and high ICS in pulmonary metastases were 10.0%, 25.5% and 47.0% (p = 0.046), respectively. Patients with high vs. low ICS in pulmonary metastases had a significantly better 5-year survival (adjusted HR 0.25, 95% CI 0.09-0.75, p = 0.013). High tumour cell PD-L1 expression in the pulmonary metastases was associated with improved survival (p = 0.024). Primary tumour CD8+ expression was significantly correlated with all T-cell densities in pulmonary metastases. Conclusion: The ICS evaluated from the resected pulmonary metastases of CRC showed significant prognostic value. High PD-L1 expression in pulmonary metastases is associated with favourable prognosis.publishedVersionPeer reviewe

Head and neck squamous cell carcinoma cell lines: Established models and rationale for selection

Background. Head and neck squamous cell carcinoma (HNSCC) cell lines are important preclinical models in the search for novel and targeted therapies to treat head and neck cancer. Unlike many other cancer types, a wide variety of primary and metastatic HNSCC cell lines are available. An easily accessible guide that organizes important characteristics of HNSCC cell lines would be valuable for the selection of appropriate HNSCC cell lines for in vitro or in vivo studies. Methods. A literature search was performed. Results. Cell growth and culture parameters from HNSCC cell lines were catalogued into tables or lists of selected characteristics. Methods for establishing cancer cell lines and basic cell culture maintenance techniques were reviewed. Conclusions. A compendium of HNSCC cell line characteristics is useful for organizing the accumulating information regarding cell line characteristics to assist investigators with the development of appropriate preclinical models. © 2006 Wiley Periodicals, Inc. Head Neck, 2006Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/55902/1/20478_ftp.pd

Les Six : Draamakasvatuksen työtavoin toteutettu lapsilähtöinen konsertti

Projektissamme kehitimme draamakasvatuksen työtapoja soveltavan lapsilähtöisen toimintamallin musiikkiopistoon. Toteutimme konsertin, jota valmistellessamme hyödynsimme draamakasvatuksen työtapoja. Konsertin teema oli Les Six, joka on 1900-luvun alkupuolella toiminut kuuden ranskalaisen säveltäjän ryhmä. Projektissamme integroimme musiikin historian ja soiton opetuksen. Piano-oppilaat saivat oman kamarimusiikkiprojektin, jossa he pääsivät soittamaan nelikätisesti ja kahdella pianolla. He pääsivät myös musisoimaan ammattilaisten kanssa, sillä myös me soitimme konsertissa. Projektissamme uudistimme siis myös musiikkiopiston konserttikäytäntöä. Halusimme ottaa projektiimme lapsilähtöisen näkökulman. Keräsimme lasten tietoja Les Six- ryhmästä ennen ja jälkeen projektin eläytymismenetelmän avulla. Heidän ajatuksiaan konserttiin valmistautumisesta kartoitimme sadutusmenetelmällä. Projekti toteutettiin Vakka-Suomen musiikkiopistolla lukukauden 2012–2013 aikana. Syyslukukauden alussa aloimme harjoitella konserttiohjelmaa. Konsertissa esitettiin Les Six- ryhmästä kertova näytelmä, jonka käsikirjoituksen kirjoitimme syksyn aikana. Keväällä 2013 pidettiin oppilaille kamarimusiikkiharjoituksia sekä kolmet harjoitukset, joissa harjoiteltiin näytelmää ja tutustuttiin Les Six- ryhmään draamakasvatuksen työtapoja hyödyntäen. Projekti huipentui konserttiin 9.3.2013. Sadutus- ja eläytymismenetelmätehtävillä keräämiemme tulosten mukaan lapset muistivat hyvin projektin aikana oppimansa musiikin historian. He myös nauttivat projektista. Havain-tojemme mukaan ryhmäytyminen onnistui hyvin. Oppilaat ottivat ennakkoluulottomasti osaa kaikkiin draamakasvatuksellisiin harjoitteisiin. Ulkopuoliselta arvioitsijaltamme saamamme palautteen mukaan tällainen projekti kannattaisi toteuttaa myös muissa musiikkiopistoissa. Työmme tarjoaa valmiin mallin Les Six- teemalla toteutettavaan konserttiin ja on myös sovellettavissa joko kokonaan tai osittain muihin teemoihin.In our development project, we created a new approach to instrumental teaching at a music institute. Our project consists of a chamber music and drama project for piano students. The theme of our project was Les Six, which was a group of six French composers of the early 1900s. We combined four aspects in our project. First, we integrated music history with piano playing by using drama pedagogy. Second, the pianists had an opportunity to play chamber music with two pianos and piano four hands. Third, we played with the students, because we wanted to give them the possibility to play in a concert with professional pianists. Fourth, we created a new approach to producing concerts at music institutes. Our project was child centered. We included the student´s point of view in our project and gathered information about their knowledge and thoughts by the storytelling method and passive drama playing. The project took place at Vakka-Suomi Music Institute. We started practicing the concert program in autumn 2012. We also wrote the play, which is based on historical facts, and planned the drama rehearsals. In spring 2013, the students had chamber music rehearsals and three music and drama rehearsals. The concert was held on 9 March 2013. According to the interviews and written stories, the students remembered the music history they learned during the project quite well. In addition, the students enjoyed the project. According to our own observations, the students formed a very good team. Even if many of the drama pedagogical exercises were new to the students, they took part in everything and seemed to have fun. Moreover, we gained feedback that this kind of a project should be carried out at other music institutes, as well. To conclude, our thesis could be used as a guide book when planning a similar project, or modify the drama pedagogical and musical elements of our project for a variety of projects.Työhön kuuluu esittelyvideo, joka on katsottavissa osoitteessa http://youtu.be/bNliiWxRaP

Immune contexture of colorectal cancer and matched liver and lung metastases

Kolorektaalisyöpä on maailman kolmanneksi yleisin syöpä ja sen alttius levitä muihin kudoksiin muodostamalla metastaaseja hankaloittaa hoitoa ja heikentää potilaan ennustetta. Elimistön immuunipuolustusjärjestelmä pyrkii eliminoimaan syöpäsoluja erityisesti T-lymfosyyttien avulla. Terveessä kudoksessa on myös Tlymfosyyttien liiallista toimintaa rajoittavia immuunivasteen tarkastuspisteitä, kuten PD-1 ja PD-L1. Syövässä tarkastuspisteiden ilmeneminen on kuitenkin yleensä normaalia suurempaa, minkä vuoksi syöpien hoitoon on kehitteillä Tlymfosyyttien toimintaa tehostavia immuunivasteen tarkastuspisteen estäjälääkkeitä. Lääkkeiden toimivuuden ja potilaan ennusteen arvioimiseksi on tärkeä selvittää syövän immunologinen tila, joka vaihtelee paljon syövän ja yksittäisten tuumorien välillä. Tutkimuksessa selvitettiin kolorektaalisyövän primäärikasvainten (N=114) sekä niiden maksa- ja keuhkometastaasien Tlymfosyyttitiheyttä ja PD-1/PD-L1-positiivisuutta sekä niiden vaikutusta potilaan ennusteeseen immunohistokemiallisesti värjättyjen tuumorinäytteiden avulla. Tlymfosyyttitiheyden huomattiin olevan korkeampi metastaaseissa kuin primäärikasvaimissa. Lisäksi keuhkometastaaseissa todettiin olevan eniten PD-1- ja PD-L1-positiivisia soluja. Aiemmista tuloksista poiketen ei korkealla Tlymfosyyttitiheydellä eikä PD-1-positiivisuudella ollut vaikutusta potilaan ennusteeseen. Sen sijaan keuhkometastaasien korkea PD-L1-positiivisuus oli yhteydessä potilaan huonompaan selviytymiseen.Colorectal cancer (CRC) is the third most common cancer type worldwide and its susceptibility to metastasize worsens the prognosis of CRC patients. Immune system strives to eliminate cancer cells mainly with tumor-infiltrating lymphocytes, such as CD4 and CD8 T lymphocytes. In healthy tissue, the excessive immune response of T lymphocytes is naturally depleted by immune checkpoints, such as PD-1 and PD-L1. In cancer the immune checkpoint expression is elevated and by targeting immune checkpoints with immune checkpoint inhibitors, their immunosuppressive function can be prevented. High density of T lymphocytes has been associated with better prognosis, whereas the effect of immune checkpoints has shown divergent results. The aim of this study was to investigate the density of T lymphocytes and the expression of PD-1 and PD-L1 both in primary CRC tumors and in their liver and lung metastases as well as their impact on the survival. The study cohort included 114 primary CRC patients with 91 liver and/or 41 lung metastases. The density of T lymphocytes and PD-1/PD-L1 expression were analyzed from immunohistochemically stained tumor samples. The results showed that T lymphocyte density was higher in metastases than in primary tumors. PD-1 and PD-L1 expressions were the highest in lung metastases. Unlike expected, higher density of T lymphocytes and stronger immune checkpoint expression were not associated with the survival of the patient