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Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the Western world covering a range from hepatic steatosis to steatohepatitis (NASH) with or without fibrosis. In a substantial number of patients, steatosis progresses to NASH, which is characterized by inflammation and cell death. Fibrosis increases risk of cirrhosis and overall mortality. NAFLD predisposes to hepatocellular carcinoma. NASH and fibrosis can be diagnosed in liver biopsy. However, non-invasive transient elastography can be used to measure liver stiffness as an estimate for fibrosis. We assessed which physical and biochemical variables associated with liver stiffness measured with transient elastography in 89 subjects. Homeostasis model assessment of insulin resistance (HOMA-IR) had the strongest association with increased liver stiffness in both linear and logistic regression models. It could easily be monitored in patients and used to get initial information on increased liver stiffness. In the literature review, I searched studies examining factors predicting advanced liver fibrosis and evaluated studies that had assessed the accuracy of transient elastography as a non-invasive method.

Pään ja kaulan alueen syövät ovat seitsemänneksi yleisin söpäryhmä. Vuosittain siihen sairastuu 660 000 ja menehtyy 325 000 henkilöä maailmanlaajuisesti. Sen merkittävimmät riskitekijät ovat ikä, tupakointi, alkoholin kuluttaminen sekä HPV-infektio. Sen esiintyvyys on merkittävässä nousussa HPV-infektioiden yleistyessä sekä väestön ikääntymisen myötä. Gerastania on olennainen tekijä potilaan hoidon ja ennusteen kannalta. Siitä huolimatta tätä aihealuetta ja sen yhteyttä pään ja kaulan alueen syöpiin on tutkittu niukasti, sen merkittävyydestä huolimatta. Tällä hetkellä potilaan hoidon- ja ennusteenarvio on yksittäisen tai muutaman kliinikon standardoimattoman arvion käsissä. Tässä narratiivisessa kirjallisuuskatsauksessa perehdytään gerasteniaan ja pään ja kaulan alueen syöpien väliseen yhteyteen kokoamalla PubMed-tietokannasta aihealuetta käsitteleviä artikkeleita hyödyntämällä avainsanoja ’Head and Neck Neoplasms’, ’Frailty’ sekä ’Complications’ sekä ’Therapeutics’. Avainsana ’Therapeutics’ viittaa siis hoitoon, joka pään ja kaulan alueen syövissä on pääosin käsittää kirurgiaa sekä kemosädehoitoa. Tutkimuskysymystä käsitteleviä löydettiin 297 kappaletta, joista 66 hyödynnettiin tutkielman kirjoittamiseen. Tutkielma osoittaa, että pään ja kaulan alueen syöpiin ja niiden hoitoon liittyy monenlaisia komplikaatioita, joilla on osoitetusti huono ennustevaikutus erityisesti gerastenian kanssa. Tutkimus myös osoittaa, että on toivottavaa arvioida systemaattisesti potilaiden gerasteniaa kvantitatiivisesti, sillä pään ja kaulan alueen syöpien potilaat ovat usein hauraita potilaita, joiden kohdalla gerastenian huomioimattomuus voi johtaa väärään tai kohtalokkaaseen hoitoon.

Amyotrophic lateral sclerosis (ALS) is the most common adult-onset motoneuron disease. ALS is characterized by a progressive loss of upper and lower motoneurons, resulting in muscle atrophy, paralysis and ultimately in death. Approximately 30,000 people die of ALS annually. There is no cure for ALS, and only two drugs - riluzole and edavarone - have been approved for the treatment of the disease. The complex pathology of ALS contributes to the lack of effective treatments. Several cellular pathologies have been suggested to contribute to the pathogenesis, including ER stress, disruption of calcium homeostasis, oxidative stress and excitotoxicity. Here we describe the cytoprotective effects of C-terminal fragments of the novel proteins with neurotrophic factor properties MANF (mesencephalic astrocyte-derived neurotrophic factor) and CDNF (cerebral dopamine neurotrophic factor) on a toxin model of ALS in vitro. Unlike the classical neurotrophic factors, MANF and CDNF are predominantly localized to the endoplasmic reticulum (ER) and have been shown to alleviate ER stress by keeping the unfolded protein response (UPR) transducers inactive. ER stress is a major component in many neurodegenerative diseases, including ALS, and is a promising therapeutic target for MANF and CDNF. However, the potential of these proteins in ALS treatment remains to be insufficiently described. We used differentiated motoneuron-like NSC-34 cells treated with a range of toxins, modelling different cellular pathologies linked to ALS. After the toxin addition, we treated the cells with MANF and CDNF variants and riluzole and measured the cell viability. The toxin panel consists of tunicamycin, ionomycin and staurosporine. Tunicamycin causes cell death by activating proapoptotic branches of the UPR. Ionomycin is an ionophore and depletes the ER of calcium, thus inducing both UPR-dependent and UPR-independent apoptosis. Less is known about the mechanisms of staurosporine, but it has been shown to induce caspase-3-dependent apoptosis, increase intracellular calcium levels and cause oxidative stress. We hypothesized that both MANF and CDNF variants protect the cells against UPR-dependent apoptosis but not against UPR-independent cell death. We show that MANF and CDNF variants protect the cells against apoptosis induced by tunicamycin, ionomycin and staurosporine. Interestingly, the protein variants mediated the highest protection against ionomycin-induced stress, and they exhibited mild protective effects against staurosporine as well. These findings suggest that MANF and CDNF variants might have a role in maintaining intracellular calcium homeostasis. However, it is possible that staurosporine induces ER stress as well, which would explain the protection conferred by the protein variant. We report that the CDNF variant mediates higher protection at lower concentrations compared to the MANF variant in every toxin assay, whereas the MANF variant mediates higher protection at the highest tested concentration compared to the CDNF variant. We also show that the CDNF variant-mediated protection against staurosporine-induced stress peaked at lower concentrations, and the highest concentration provided distinctively lower, yet significant effect. These data lead us to hypothesize that the protein variants may have a slightly different mode of action, and that they might provide an additive effect when administered simultaneously. We tested a combination of MANF and CDNF variants in cells treated with tunicamycin, ionomycin and staurosporine. However, the combination treatment did not increase the viability more than MANF and CDNF variants independently did. The results answered our questions as well as raised new ones. In the future, the putative calcium-regulating effects of the protein variants should be investigated. The UPR-modifying effects of the drug candidates and toxins need to be assessed by quantifying changes in the UPR marker mRNA and protein expression levels. If it is revealed that the variants have a different mode of action, the possible additive protective effects must be assessed. Finally, a wider toxin panel is needed to fully explore the potential of MANF and CDNF variants in ALS treatment. This study demonstrates the potential of MANF and CDNF variants in protecting motoneurons against several pathological pathways contributing to ALS pathology. However, the mechanisms of action of the variants need further investigation to fully understood their therapeutic potential.

The aim of the study. Spontaneous eye blink rate (sEBR) is a behavioral index that has been linked to frontostriatal dopaminergic activity. Reduced or increased dopaminergic activity due to clinical conditions tends to be associated with lower or higher sEBR, respectively, and sEBR can be modulated by pharmacological agents that affect dopamine signaling. Consequently, sEBR could serve as an easily accessible method of assessing brain dopaminergic tone indirectly in humans. It might be preferable to more expensive and invasive techniques such as positron emission tomography. However, it remains unclear whether variations in dopaminergic genes predict sEBR. In this cross-sectional study, the relationship between sEBR and dopaminergic genotype was examined in two samples. Two genetic polymorphisms were focused on: the COMT Val158Met polymorphism and the A1 allele of the Taq1A polymorphism. It was hypothesized that the COMT Val158Met polymorphism is associated with higher sEBR, and that the A1 allele of the Taq1A polymorphism is associated with lower sEBR. As BMI and diet have been linked with altered striatal dopamine function, the possible association between BMI, diet, and sEBR was studied exploratively. Methods. Data from three cross-sectional studies was used in this study: The intervention study (n = 31) is an experimental study that examines the effect of acute phenylalanine/tyrosine depletion on cognitive measures. The GREADT study (n = 86) focuses on the effects of genotype and diet on dopamine tone. The BEDOB study (n = 69) investigates neurocognitive mechanisms in obesity and binge eating disorder. Similar methodologies were used in the GREADT and the BEDOB studies, which is why these datasets were combined for the analyses. Blink rates were measured using an infrared eye tracking system. The participants completed the Dietary Fat and free Sugar Questionnaire (DFS) to assess how much they consumed saturated fat and refined sugar. In the GREADT/BEDOB sample, the associations between the polymorphisms, BMI, DFS-score, and sEBR were examined with univariate analyses of variance. In the intervention study sample, a generalized linear mixed model was run to check whether sEBR changed in the intervention and whether the genotypes, BMI, or DFS-score affected sEBR. Results. No influence of the genotypes was found on sEBR in either of the samples. BMI had a significant effect on sEBR in the GREADT/BEDOB sample. The association was significant in the overweight/obese group but not in the normal weight group. DFS-score did not influence sEBR in either of the samples. Conclusions. The results of this study converge with those of authors suggesting caution in using sEBR as a proxy for central dopamine functioning of healthy humans. In future studies, particular attention should be paid to methodological considerations when studying sEBR.

Summary of Background Data. Lenke classification is used to define the curve type in adolescent idiopathic scoliosis (AIS). The association of Lenke classification and long-term postoperative health-related quality of life (HRQoL) remains unclear. Objective. The purpose of this study was to assess the association between Lenke classification and HRQoL in patients who underwent spinal fusion for AIS. Materials and Methods. In all, 146 consecutive patients (mean age 15.1 yr) operated for AIS between 2007 and 2019 with a minimum 2-year follow-up were included. Fifty-three (36%) patients reached the 10-year follow-up. Their HRQoL was assessed with the SRS-24 questionnaire preoperatively, at six months, two years, and 10 years after surgery. Results. The preoperative major curve was the largest in Lenke 3 (mean 63°) and 4 (mean 62°) groups and the lowest in Lenke 5 groups (mean 48°, P<0.05). These curves were corrected to a mean of 15° with no differences between groups. We found no evidence of differences between the preoperative HRQoL scores between the Lenke groups. The self-image domain of SRS-24 was lower in patients with isolated major thoracolumbar scoliosis (Lenke 5) when compared with double-thoracic (Lenke 2) group at the two-year follow-up (mean [95% CI] 3.6 [3.3–3.9] vs. 4.3 [4.1–4.6]). The postoperative satisfaction domain was lower in Lenke 5 group when compared with main thoracic (Lenke 1) group (mean [95% CI] 3.8 [3.5–4.0] vs. 4.3 [4.2–4.5]) and Lenke 2 group (mean 4.4, 95% CI 4.2–4.6) at the two-year follow-up. The mean total score of SRS-24 at the 10-year follow-up was highest in Lenke 1 group (mean 4.06, 95% CI 3.79–4.33) and lowest in Lenke 6 group (mean 2.92, 95% CI 2.22–3.61). Conclusions. Lenke classification and especially its curve type (major thoracic vs. major thoracolumbar scoliosis) was associated with long-term health-related quality of life after instrumented spinal fusion for AIS.

Työperäinen altistuminen voi pahentaa jo olemassa olevan astman oireita tai aiheuttaa uutta työperäistä astmaa, ja sillä on osoitettu olevan myös merkitystä keuhkoahtaumataudin (COPD) synnyssä. Tämän tutkimuksen tarkoituksena oli tarkastella voivatko eroavaisuudet ammattiluokituksissa, työperäisessä altistumisessa ja tupakoinnissa vaikuttaa astman, COPD:n, hengitystieoireiden ja ilmateiden eosinofiilisen tulehduksen esiintyvyyteen. Tutkimuksessa tarkasteltiin myös kuinka eri ammattiluokittelut soveltuvat epidemiologiseen tutkimukseen. Kolmen Itämeren alueen valtion (Suomi, Viro ja Ruotsi) yhteistyönä toteutettiin vuosituhannen vaihteessa monikeskustutkimus, jossa tutkittaville tehtiin jokaisessa keskuksessa yhteneväinen lomakehaastattelu, spirometria sekä uloshengitysilman typpioksidimittaus (FENO). Tätä kolmesta maasta kerättyä aineistoa käytettiin nykyiseen tutkimukseen, jossa sitä tarkasteltiin kolmen eri ammattiluokittelun avulla. Lisäksi arvioitiin työperäistä altistumista altistematriisin (JEM) avulla. Iän, sukupuolen ja tupakoinnin mukaan säädetyssä monimuuttuja-analyysissa havaittiin tilastollisesti merkittävä riski kahdessa eri ammattiluokittelussa, jotka liittyivät ei-manuaaliseen työhön ja terveydenhoito- ja sosiaalialan töihin. Näissä ryhmissä oli myös korkea hengitystieoireiden esiintyvyys. Merkittävä COPD:n riski tuli ilmi kaikissa kolmessa luokitusjärjestelmässä manuaalisten töiden osalta ja työperäinen altistus aiheutti myös merkittävän COPD:n riskin sekä COPD:n liittyvien hengitystieoireiden riskin monimuuttuja-analyysissä. Uloshengitysilman typpioksidimittaustulokset erosivat esiintyvyyksiltään ammattiryhmissä mutta monimuuttuja-analyysissä ei havaittu merkittäviä riskejä kohonneen FENO:n suhteen. Eri ammattiluokituksilla oli erilainen kyky löytää tauteja, oireita ja työperäistä altistumista, ja tutkimuksen hyöty on auttaa sopivien työkalujen käyttämisessä tulevissa epidemiologissa tutkimuksissa, jotka tarkastelevat myös sosioekonomista näkökulmaa.

DBH-geeni koodaa dopamiinia hajottavaa entsyymiä, joka on liitetty palkkiojärjestelmään vaikuttamisen kautta erilaisiin riippuvuuksiin. DBH:n variaatioiden on todistettu liittyvän tupakointikäytänteisiin ja –tapoihin monessa tutkimuksessa. Viimeisimpänä rs3025343 liitettiin tupakoinnin lopettamiseen suuressa GWAS meta-analyysissa. Tutkimuksemme tavoitteena olikin replikoida kyseinen löydös suuressa väestötutkimusnäytteessämme. Lisäksi halusimme tutkia, vaikuttaisiko rs3025343 jonkin muun ympäristötekijän kautta vai itsenäisesti tupakoinnin lopettamiseen. Tutkimusnäytteemme on peräisin suomalaisesta väestönäytteestä, FINRISK-tutkimuksesta. Siihen lukeutuu 26,582 genotyypattua henkilöä tupakointistatuksineen. Analyysit rajasimme 11,926 yksilöön, jotka olivat joko nykyisiä tupakoitsijoita (n=6,578) tai vähintään 6kk sitten tupakoinnin lopettaneita (n=5,348). Henkilöistä oli saatavilla myös kattavasti muita tietoja mukaan lukien sosioekonominen status, terveyteen liittyviä tapoja ja terveydentila, joita käytimme analyyseissä hyväksi. Yhteys rs3025343 ja tupakoinnin lopettamisen välillä (OR=1.12, p=0.094, 95%CI=0.98-1.30) osoittautui tutkimuksessamme identtiseksi GWAS-tutkimuksen kanssa (OR=1.12, 95%CI=1.08-1.18). Mikään testatuista fenotyypeistämme ei vaikuttanut tuohon yhteyteen merkitsevästi. Siviilisääty, koulutustaso, masennus, alkoholin käyttö, itseraportoitu terveys sekä COPD assosioituivat fenotyyppitekijöistä tupakoinnin lopettamiseen, mutta mikään edellämainituista assosiaatioista ei riippunut tutkimastamme genotyypistä. Vaikka tutkimustuloksemme ei ole tilastollisesti merkitsevä, efektikoko viittaa vahvasti siihen, että tutkimallamme polymorfismilla on jonkinasteinen yhteys tupakoinnin lopettamiseen. Merkitevyyden esiinsaamiseksi riittävällä voimalla (80%) otoskoon tulisi olla niinkin suuri kuin 36,092 tapausta ja 29,343 kontrollia, koska harvinaisemman alleelin kantajia on suhteellisen vähän (7,1%). DBH-geenin variaatiot ovat osoittautuneet monessa tutkimuksessa olevan yhteydessä nikotiiniriippuvuuteen tai tupakoinnin lopettamiseen. Jos näitä yhteyksiä saadaan tutkittua lisää, on mahdollista että tietoja voitaisiin käyttää hyväksi tulevaisuudessa esim. yksilöllisesti räätälöidyssä tupakoinnin lopettamisen hoidossa.

Objectives. Recent results of both animal and human studies suggest that intestinal microbiota, i.e. microorganisms inhabiting the gastrointestinal system, may be connected to their host’s cognition. However, the diverse effects of intestinal microbiota are still poorly understood and especially knowledge of its associations with normative childhood cognitive development is very scarce. The purpose of the current study was to examine the possible associations between infant intestinal microbial composition, richness and diversity and cognitive performance in early childhood. Methods. The current study sample consisted of the children taking part in Finnish Health and Early Life Microbiota (HELMi) longitudinal birth cohort study. The cognitive abilities of 424 children were assessed at 2 years of age with Bayley Scales of Infant and Toddler Development, using cognitive, receptive language and expressive language subscales. Of 424 tested children, those from whom microbiota analysis for at least one fecal sample was available at the time of the start of this study, were included. Fecal samples were collected when infants were 3, 6 and 12 weeks old and 6, 9 and 12 months old, and the bacterial composition, richness and diversity were analyzed with 16S rRNA- amplicon sequencing method. Results and conclusions. Intestinal microbial composition in infancy was found to be related to cognitive abilities of the children, more specifically, receptive language skills and expressive language skills. A higher abundance of the genus Finegoldia at 12 weeks of age and the genus Serratia at 6 months of age were related to worse receptive language performance at 2 years of age. A higher abundance of the family Enterococcaceae at 12 weeks of age and the genus Alistipes at 6 months of age, were associated with worse expressive language skills. In addition, the children who scored in lowest 20th percentile in the receptive language tasks, had richer intestinal microbiota at 3 weeks and 6 months of age. Conclusions cannot yet be drawn based on these preliminary findings, but the results suggest that infant intestinal microbiota may be one of the factors influencing cognitive, especially verbal, development in early childhood.

Objective. Depression is associated with increased risk of chronic disease, which may be at least partly due to poor health behaviors. Growing body of evidence has associated depression with unhealthy diet. However, the association of depression with diet quality in the long run is not well known. Furthermore, it is unclear if dietary interventions could mitigate the harmful association of depression with diet. This study examined the association of depression with diet both cross-sectionally and longitudinally in a population-based prospective cohort. The effectiveness of an early-onset dietary intervention in modifying these associations was investigated. Methods. The sample (n = 457) was from The Special Turku Coronary Risk Factor Intervention Project (STRIP). The intervention group (n = 209) had undergone a dietary intervention lasting from age of 7 months until age of 20 years. Depression was measured at age 20 using Beck Depression Inventory II (BDI-II). Diet quality was assessed at ages 20 and 26 using a diet score calculated based on food diaries. Missing values were replaced using multiple imputation by chained equations. Linear regression analyses were used to analyze the association of depression at age 20 with diet at ages 20 and 26, as well as the modifying effect of intervention group on these associations. Results. No cross-sectional association was found for depression and diet at age 20. Depression at age 20 was longitudinally associated with worse diet quality at age 26. The associations did not differ between intervention and control groups at either of the time points. Conclusions. Contrary to previous research, this study did not find cross-sectional association for depression with diet. However, this study offers novel information on longitudinal associations, suggesting that depression may have effects on diet quality that can manifest after several years. Dietary intervention was not found effective in modifying these associations. Since long-term effects on diet may be an important factor explaining the association of depression with chronic diseases, ways to mitigate the adverse consequences of depression for diet should be explored further.

Immunophenotyping by flow cytometry (FC) is an established practice to identify immune cells and their cellular changes at the single-cell level. Since preserving the structural integrity of cellular epitopes is vital for immunophenotyping, samples should be processed shortly after being collected. However, the requirements of complex facilities and trained personnel for flow cytometry make it challenging to handle samples immediately. Fixation and cryopreservation extend sample shelf life and allow analysing longitudinal samples simultaneously while minimizing technical variation. Nevertheless, usage of whole blood cryopreservation in flow cytometry is limited due to challenges in preserving epitope structures during fixation and detecting dim antigens. This thesis investigates the performances of four commercial whole blood cryopreserving kits; 1) Cytodelics, 2) Stable-Lyse V2 and Stable-Store V2 (SLSS-V2), 3) Proteomic stabiliser (PROT-1), and 4) Transfix. Peripheral blood samples were processed with these stabilising buffers immediately after the collection and cryopreserved until further analysis by flow cytometry. Here, we measured the stability of major immune lineages, T cell subpopulations, and activated neutrophil profiles in samples treated with these commercial whole blood stabilisers. Our flow cytometry data showed that PROT-1, Transfix and Cytodelics maintained the distribution of major leukocyte subsets – granulocytes, T cells, natural killer cells and B cells, comparable to unpreserved samples despite the attenuation of fluorescence intensities. Moreover, these three stabilisers also preserved phenotypes of activated neutrophils upon stimulation with N-Formylmethionyl-leucyl-phenylalanine and Lipopolysaccharides. The upregulation of adhesion molecules (CD11b), Fc receptors (CD16) and granule proteins (CD66b) as well as the shedding of surface L-selectin (CD62L) on activated neutrophils was conserved most efficiently in PROT-1, followed by Cytodelics. On the other hand, none of the stabilisers provided a reliable detection of CCR7 for accurate quantification of T cell subpopulations. COVID-19 is caused by a highly transmissible and pathogenic coronavirus, so-called severe acute respiratory syndrome coronavirus 2 (SARS-COV-2). To test the potential of whole blood cryopreservation kits for flow cytometry in COVID-19 research, we studied the detectability of major leukocyte lineages and granulocyte subsets in longitudinal patient samples processed with Cytodelics. High dimensional analysis with Uniform Manifold Approximation and Projection (UMAP) and Self-Organising Maps (FlowSOM) clustering revealed remarkable stability of CD3, CD15, and CD14 expression in samples stored with Cytodelics. It allowed the detection of lymphopenia and emergency granulopoiesis often found during the acute phase of severe SARS-COV-2 infection. Nonetheless, we could not determine signatures of granulocyte subsets, notably suppressive neutrophils, during the acute and convalescent phases of COVID-19. Variable detection of lowly expressed markers and diminished fluorescence intensities in Cytodelics - preserved samples might have hindered the analysis. In conclusion, this study demonstrates that PROT-1, Transfix, and Cytodelics enabled reliable detection of highly expressed leukocyte markers, whereas SLSS-V2 preservation resulted in the most inaccurate identification of studied markers. Notably, our results show that Cytodelics can be applied in COVID-19 studies to immunophenotype major immune lineages by flow cytometry. Nevertheless, more optimisation is needed for less abundant or fixation-sensitive epitopes to enhance the efficacy of whole blood cryopreservation for flow cytometry.