Microtubule-mediated protein transport mechanisms during spermiogenesis

Formation of cilia and flagella is a tightly controlled process organized by intraflagellar transport (IFT), for which two motor proteins, kinesin-2 and dynein, are responsible. The exact functions of IFT and IFT-related proteins in spermiogenesis are poorly understood in mammalian species. To investigate mechanisms of IFT during mouse spermatogenesis, a male germ cell-specific knockout mouse model for kinesin-2 subunit kinesin-like protein 3A (KIF3A) was generated. Depletion of KIF3A caused defects in sperm tail development and the core structure, the axoneme, failed to form. A transient microtubular platform, the manchette, surrounds the sperm head during spermiogenesis. In KIF3A-depleted mice, the manchette appeared constricted and its clearance was delayed, causing abnormal head shape, suggesting that IFT and KIF3A function are also important for manchette function. In addition, we identified the KIF1-binding protein (KBP) and the meiosis-specific nuclear structural protein 1 (MNS1) as a novel interaction candidates for KIF3A. Sperm flagellar protein 2 (SPEF2) has been shown to localize in elongating spermatids, and its interaction with IFT-related protein IFT20 has been established. To characterize the function of SPEF2 in sperm tail formation, a male germ cell-specific conditional knockout mouse model for SPEF2 was generated (Spef2 cKO). Depletion of SPEF2 caused defects in axoneme formation, and similar defects in the manchette and head shaping were observed in KIF3A-depleted mice. We also identified cytoplasmic dynein 1 and GOLGA3 as novel interaction candidates for SPEF2 in the testis. Inhibition of dynein 1 activity in the manchette blocked the SPEF2 transport, suggesting its role in manchette-related transport. IFT20 and SPEF2 localization in the Golgi complex and the delayed appearance of IFT20 in the manchette in Spef2 cKO suggests that SPEF2 functions as an adaptor in dynein 1-mediated transport in elongating spermatids. This study increases our understanding of the protein transport mechanisms required for the formation of functional spermatozoa.Mikrotubulusvälitteiset proteiinien kuljetusmekanismit spermiogeneesin aikana Solunsisäisen kuljetusmekanismin (intraflagellar transport, IFT) oikeanlainen toiminta on edellytys värekarvan ja siittiön hännän muodostumiselle. Kuljetusmekanismissa toimivat moottoriproteiinit, kinesiini-2 ja dyneiini ovat vastuussa proteiinien kuljetuksesta. Kuljetusmekanismin toiminta siittiön hännän muodostuksen aikana on vielä melko tuntematon. Tässä tutkimuksessa olemme pyrkineet selvittämään kuljetusmekanismin toimintaa poistamalla kinesiini-2 proteiinin alayksikön kinesiinin-kaltainen proteiini 3A:n (KIF3A) urosten sukusoluista. KIF3A proteiinin poisto aiheutti ongelmia siittiön hännän muodostuksen aikana. Siittiön hännän ydinrakenne, aksoneema, ei muodostunut sekä siittiön pään ympärille hetkellisesti ilmentyvä mikrotubulusrakenne, mansetti, oli epämuodostunut sekä sen poistuminen viivästynyt. Tunnistimme myös KIF3A:n kanssa toimivia proteiineja kiveksessä kuten KIF1:tä sitovan proteiinin (KBP) sekä meioosi-spesifin tuman rakenne proteiinin 1:n (MNS1). Siittiön flagella proteiini 2:n (SPEF2) on näytetty ilmentyvän siittiön hännän muodostumisen aikana. Sen toiminta IFT kuljetusmekanismiin liittyvän IFT20 proteiinin kanssa on osoitettu aiemmissa tutkimuksissa. Tässä tutkimuksessa teimme urosten sukusoluspesifisen poistogeenisen hiirimallin SPEF2:lle ymmärtääksemme paremmin sen toimintaa siittiön hännän muodostumisen aikana. SPEF2 proteiinin poisto aiheutti ongelmia aksoneeman muodostuksessa, mansetti oli epämuodostunut sekä poisto viivästynyt, samoin kuten havaitsimme KIF3A poistogeenisellä hiirellä. Tunnistimme GOLGA3:n ja sytoplasmisen dyneiini 1:n SPEF2:n kanssa toimiviksi proteiineiksi. Osoitimme myös että dyneiini 1 moottorin toiminnan häiriö estää SPEF2 proteiinin kuljetuksen mansetissa. SPEF2 ja IFT20 proteiinien sijoittuminen Golgiin sekä IFT20 proteiinin viivästynyt paikantuminen mansettiin SPEF2 poistogeenisellä hiirellä viittaavat SPEF2 proteiinin mahdollisesta roolista toimia linkkinä dyneiini 1 moottorin ja IFT20 proteiinin välillä. Tämä tutkimus auttaa ymmärtämään proteiinien kuljetusmekanismeja siittiön kehityksen aikanaSiirretty Doriast

Impedanssitomografia sovellettuna kantasoluihin hydrogeeli skaffoldissa

Electrical impedance tomography (EIT) could provide a label free, non-invasive and fast method for cell culture monitoring. This thesis is a feasibility study on what kind of results EIT may provide when cells are cultured in a 3D hydrogel scaffold. The work is divided into two parts: (1) electrical impedance spectroscopy (EIS) measurements and (2) computer modelling of the EIT setup. In the EIS study part, gellan gum (GG) hydrogels and GG hydrogels with encapsulated stem cells are measured using the impedance spectroscope HF2IS device (Zurich Instruments AG, Switzerland). In the second study part, two dimensional EIT computer models are done in COMSOL Multiphysics. The sensitivity field of EIT setup is simulated in order to find the optimal electrode locations and resistivity values for the aqueous solution used between electrodes and the sample. The EIS experiments indicate that the impedance value is dependent on the amount and viability of cells. Optimal electrode configuration and resistivity values are obtained by the EIT model and these parameters can be used in the future EIT measurements

A novel mutation in the matrix metallopeptidase 2 coding gene associated with intrafamilial variability of multicentric osteolysis, nodulosis, and arthropathy

Background MONA, which stands for a spectrum of Multicentric Osteolysis, subcutaneous Nodulosis, and Athropathia, is an ultra rare autosomal recessive disorder caused by mutations in the matrix metallopeptidase 2 (MMP2) gene. To date only 44 individuals, carrying 22 different mutations have been reported. Here we report on two brothers with identical homozygous MMP2 gene mutations, but with clearly different phenotypes. Methods Genomic DNA was isolated from the affected brothers and the parents. An iliac crest bone biopsy was taken from the younger patient (index case). The level of matrix metallopeptidase 2 enzyme (MMP2) in serum and synovial fluid of the younger patient was analyzed using gelatin zymography. Results The DNA analysis revealed a homozygous c.1188C>A transversion on exon 8 of the gene. The affected brothers had the same homozygous variant and the parents were heterozygous to this variant. This variant has been reported as a compound heterozygous mutation on one individual resulting in scleroderma like skin thickening. Bone histomorphometry indicated increased trabecular bone remodeling and turnover. The zymography revealed that the level of MMP2 was completely nonmeasurable in the serum and only a minor gelatinolytic protein band of about similar molecular weight as MMP2 was found in the synovial fluid. Conclusions Both the age at the onset and the phenotypic severity of the syndrome in these two brothers were different despite identical genotypes. The younger patients had corneal opacities leading to deteriorating visual acuity. For the first time in this disease, opacities were successfully treated with corneal transplantations.Peer reviewe

A splice site variant in INPP5E causes diffuse cystic renal dysplasia and hepatic fibrosis in dogs

Ciliopathies presenting as inherited hepatorenal fibrocystic disorders are rare in humans and in dogs. We describe here a novel lethal ciliopathy in Norwich Terrier puppies that was diagnosed at necropsy and characterized as diffuse cystic renal disease and hepatic fibrosis. The histopathological findings were typical for cystic renal dysplasia in which the cysts were located in the straight portion of the proximal tubule, and thin descending and ascending limbs of Henle’s loop. The pedigree of the affected puppies was suggestive of an autosomal recessive inheritance and therefore, whole exome sequencing and homozygosity mapping were used for identification of the causative variant. The analyses revealed a case-specific homozygous splice donor site variant in a cilia related gene, INPP5E: c.1572+5G>A. Association of the variant with the defect was validated in a large cohort of Norwich Terriers with 3 cases and 480 controls, the carrier frequency being 6%. We observed that the identified variant introduces a novel splice site in INPP5E causing a frameshift and formation of a premature stop codon. In conclusion, our results suggest that the INPP5E: c.1572+5G>A variant is causal for the ciliopathy in Norwich Terriers. Therefore, genetic testing can be carried out in the future for the eradication of the disease from the breed

Suomen zoonoositilanne ja riskit yhteisen terveyden näkökulmasta : Yhteenveto zoonoosien suuntauksista ja lähteistä 2011–2021

Kampylobakteerit ovat yleisin ja salmonellat toiseksi yleisin ihmisillä suolistoinfektioita aiheuttavat zoonoosit Suomessa. Kryptosporidioosi- ja shigatoksinen Eshericia coli (STEC) -tapauksia on ilmoitettu aikaisempaa enemmän. Vektorivälitteisistä zoonooseista borrelioosi on Suomessa yleisimmin todettu. Puutiaisaivotulehdustapausten määrä on noussut. Tuotantoeläimistä salmonellatapaukset naudoilla ja sioilla lisääntyivät, kun taas broilereilla ne vähenivät. Salmonellaa esiintyy kotimaisessa naudan- ja sianlihassa edelleen vähän. Teurasbroilereilla kampylobakteerien esiintyvyys on vähentynyt. Teurasnaudoissa STEC O157 -bakteerit yleistyivät. Cryptosporidium parvum -tautitapaukset nautakarjoissa lisääntyivät voimakkaasti. Trikinellatartunnat sioilla harvinaistuivat, eikä tartuntoja sikaloissa ole viime vuosina todettu. Metisilliiniresistentti Staphylococcus aureus yleistyi teurassioissa. Luonnonvaraisilla linnuilla todettiin Suomessa ennen havaitsematon korkeapatogeeninen lintuinfluenssa. Elintarvikkeista raakamaito toimi useiden suolistoinfektioita aiheuttavien zoonoosien välittäjäelintarvikkeena, lisäksi kasvisten merkitys zoonoosien välittäjänä korostui. Suomen zoonositilanteeseen vaikuttavat ympäristöön ja ilmastoon liittyvät muutokset, vastustuskyvyltään heikentyneiden henkilöiden määrä, sekä joidenkin zoonoosien osalta endeemisiltä aluilta Suomeen tuotavat koirat. Esitettävät tiedot auttavat suuntaamaan ennakoivia toimia Suomen zoonoositilanteen säilyttämiseksi hyvänä ja tunnistamaan ne kohteet, joihin on syytä panostaa.Tämä julkaisu on toteutettu osana valtioneuvoston selvitys- ja tutkimussuunnitelman toimeenpanoa. (tietokayttoon.fi) Julkaisun sisällöstä vastaavat tiedon tuottajat, eikä tekstisisältö välttämättä edusta valtioneuvoston näkemystä

Genes reveal traces of common recent demographic history for most of the Uralic-speaking populations

Background: The genetic origins of Uralic speakers from across a vast territory in the temperate zone of North Eurasia have remained elusive. Previous studies have shown contrasting proportions of Eastern and Western Eurasian ancestry in their mitochondrial and Y chromosomal gene pools. While the maternal lineages reflect by and large the geographic background of a given Uralic-speaking population, the frequency of Y chromosomes of Eastern Eurasian origin is distinctively high among European Uralic speakers. The autosomal variation of Uralic speakers, however, has not yet been studied comprehensively.Results: Here, we present a genome-wide analysis of 15 Uralic-speaking populations which cover all main groups of the linguistic family. We show that contemporary Uralic speakers are genetically very similar to their local geographical neighbours. However, when studying relationships among geographically distant populations, we find that most of the Uralic speakers and some of their neighbours share a genetic component of possibly Siberian origin. Additionally, we show that most Uralic speakers share significantly more genomic segments identity-by-descent with each other than with geographically equidistant speakers of other languages. We find that correlated genome-wide genetic and lexical distances among Uralic speakers suggest co-dispersion of genes and languages. Yet, we do not find long-range genetic ties between Estonians and Hungarians with their linguistic sisters that would distinguish them from their non-Uralic-speaking neighbours.Conclusions: We show that most Uralic speakers share a distinct ancestry component of likely Siberian origin, which suggests that the spread of Uralic languages involved at least some demic component

Genes reveal traces of common recent demographic history for most of the Uralic-speaking populations.

BACKGROUND: The genetic origins of Uralic speakers from across a vast territory in the temperate zone of North Eurasia have remained elusive. Previous studies have shown contrasting proportions of Eastern and Western Eurasian ancestry in their mitochondrial and Y chromosomal gene pools. While the maternal lineages reflect by and large the geographic background of a given Uralic-speaking population, the frequency of Y chromosomes of Eastern Eurasian origin is distinctively high among European Uralic speakers. The autosomal variation of Uralic speakers, however, has not yet been studied comprehensively. RESULTS: Here, we present a genome-wide analysis of 15 Uralic-speaking populations which cover all main groups of the linguistic family. We show that contemporary Uralic speakers are genetically very similar to their local geographical neighbours. However, when studying relationships among geographically distant populations, we find that most of the Uralic speakers and some of their neighbours share a genetic component of possibly Siberian origin. Additionally, we show that most Uralic speakers share significantly more genomic segments identity-by-descent with each other than with geographically equidistant speakers of other languages. We find that correlated genome-wide genetic and lexical distances among Uralic speakers suggest co-dispersion of genes and languages. Yet, we do not find long-range genetic ties between Estonians and Hungarians with their linguistic sisters that would distinguish them from their non-Uralic-speaking neighbours. CONCLUSIONS: We show that most Uralic speakers share a distinct ancestry component of likely Siberian origin, which suggests that the spread of Uralic languages involved at least some demic component

Genomic analyses inform on migration events during the peopling of Eurasia.

High-coverage whole-genome sequence studies have so far focused on a limited number of geographically restricted populations, or been targeted at specific diseases, such as cancer. Nevertheless, the availability of high-resolution genomic data has led to the development of new methodologies for inferring population history and refuelled the debate on the mutation rate in humans. Here we present the Estonian Biocentre Human Genome Diversity Panel (EGDP), a dataset of 483 high-coverage human genomes from 148 populations worldwide, including 379 new genomes from 125 populations, which we group into diversity and selection sets. We analyse this dataset to refine estimates of continent-wide patterns of heterozygosity, long- and short-distance gene flow, archaic admixture, and changes in effective population size through time as well as for signals of positive or balancing selection. We find a genetic signature in present-day Papuans that suggests that at least 2% of their genome originates from an early and largely extinct expansion of anatomically modern humans (AMHs) out of Africa. Together with evidence from the western Asian fossil record, and admixture between AMHs and Neanderthals predating the main Eurasian expansion, our results contribute to the mounting evidence for the presence of AMHs out of Africa earlier than 75,000 years ago.Support was provided by: Estonian Research Infrastructure Roadmap grant no 3.2.0304.11-0312; Australian Research Council Discovery grants (DP110102635 and DP140101405) (D.M.L., M.W. and E.W.); Danish National Research Foundation; the Lundbeck Foundation and KU2016 (E.W.); ERC Starting Investigator grant (FP7 - 261213) (T.K.); Estonian Research Council grant PUT766 (G.C. and M.K.); EU European Regional Development Fund through the Centre of Excellence in Genomics to Estonian Biocentre (R.V.; M.Me. and A.Me.), and Centre of Excellence for Genomics and Translational Medicine Project No. 2014-2020.4.01.15-0012 to EGC of UT (A.Me.) and EBC (M.Me.); Estonian Institutional Research grant IUT24-1 (L.S., M.J., A.K., B.Y., K.T., C.B.M., Le.S., H.Sa., S.L., D.M.B., E.M., R.V., G.H., M.K., G.C., T.K. and M.Me.) and IUT20-60 (A.Me.); French Ministry of Foreign and European Affairs and French ANR grant number ANR-14-CE31-0013-01 (F.-X.R.); Gates Cambridge Trust Funding (E.J.); ICG SB RAS (No. VI.58.1.1) (D.V.L.); Leverhulme Programme grant no. RP2011-R-045 (A.B.M., P.G. and M.G.T.); Ministry of Education and Science of Russia; Project 6.656.2014/K (S.A.F.); NEFREX grant funded by the European Union (People Marie Curie Actions; International Research Staff Exchange Scheme; call FP7-PEOPLE-2012-IRSES-number 318979) (M.Me., G.H. and M.K.); NIH grants 5DP1ES022577 05, 1R01DK104339-01, and 1R01GM113657-01 (S.Tis.); Russian Foundation for Basic Research (grant N 14-06-00180a) (M.G.); Russian Foundation for Basic Research; grant 16-04-00890 (O.B. and E.B); Russian Science Foundation grant 14-14-00827 (O.B.); The Russian Foundation for Basic Research (14-04-00725-a), The Russian Humanitarian Scientific Foundation (13-11-02014) and the Program of the Basic Research of the RAS Presidium “Biological diversity” (E.K.K.); Wellcome Trust and Royal Society grant WT104125AIA & the Bristol Advanced Computing Research Centre (http://www.bris.ac.uk/acrc/) (D.J.L.); Wellcome Trust grant 098051 (Q.A.; C.T.-S. and Y.X.); Wellcome Trust Senior Research Fellowship grant 100719/Z/12/Z (M.G.T.); Young Explorers Grant from the National Geographic Society (8900-11) (C.A.E.); ERC Consolidator Grant 647787 ‘LocalAdaptatio’ (A.Ma.); Program of the RAS Presidium “Basic research for the development of the Russian Arctic” (B.M.); Russian Foundation for Basic Research grant 16-06-00303 (E.B.); a Rutherford Fellowship (RDF-10-MAU-001) from the Royal Society of New Zealand (M.P.C.)

Alaraaja-ateroskleroosi potilaan ohjaus sairaanhoitajan näkökulmasta

Tämän työn teoreettinen osio koostuu ateroskleroosin kuvauksesta, sen syntyyn vaikuttavista tekijöistä sekä hoidollisesta osiosta, ohjauksen, vuorovaikutuksen ja hoitoon sitoutumisen merkityksestä potilasohjauksessa. Ateroskleroosilla (ASO) tarkoitetaan valtimonkovettumatautia. Sitä esiintyy aortassa sekä suurissa että pienissä valtimoissa. ASO:ssa aortan alavartaloon vievä osa tai sen haara ahtautuu. Alaraaja-ASO voi johtaa iskemiaan. Alaraaja-ASO:n syntyyn vaikuttavat tupakointi, diabetes, koholla olevat sokeriarvot sekä verenkiertohäiriöt. Asiantuntijuuteen perustuva tieto nykyään kyseenalaistetaan. Asiakkaan ja terveysalan ammattilaisen välillä tapahtuva terveyden edistäminen perustuu vuorovaikutukseen. Hoitoon sitoutumisella tarkoitetaan asiakkaan vastuullista ja aktiivista toimintaa terveyden edellyttämällä tavalla. Opinnäytetyömme tarkoituksena oli auttaa sairaanhoitajia kehittämään omaa ohjaustyötään alaraaja-ASO potilaiden hoidossa. Työ toteutettiin laadullisena tutkimuksena. Aineiston keräsimme haastattelemalla neljää sairaanhoitajaa, jotka työskentelevät alaraaja-ASO potilaiden kanssa. Haastattelu sisälsi kolme eri teemaa; ohjaus, vuorovaikutus ja hoitoon sitoutuminen. Työssä käytimme aineistolähtöistä sisällönanalyysiä (n=4), jossa lähtökohtana olivat haastateltavien sairaanhoitajien alkuperäiset ilmaukset. Haastattelut nauhoitettiin, jonka jälkeen ne litteroitiin eli kirjoitettiin sanasta sanaan. Tuloksissa kävi ilmi, kuinka tärkeää on ottaa potilaan ohjauksessa huomioon riskitekijät. Suurimpana riskitekijänä tuli esiin tupakka. Vuorovaikutustilanteeseen vaikuttaa suuresti potilaan ikä. Vanhusten ohjaus koettiin ajoittain haasteelliseksi. Suurimpana haasteena ohjauksessa sairaanhoitajat kokivat ajanpuutteen. Aikaa ohjaukseen pitäisi olla enemmän, jotta ohjaus olisi tarpeeksi riittävää. Työn tarkoitus on, että sen avulla sairaanhoitajat saavat uutta tietoa ohjauksesta ja vuorovaikutuksesta potilaiden kanssa sekä saavat lisää teoreettista tietoa ohjauksesta, vuorovaikutuksesta sekä hoitoon sitoutumisesta. Valmis työ toimitettiin Meilahden sairaalan kirurgian poliklinikalle, jossa työskennellään alaraaja-ASO potilaiden kanssa.Nurses’ view of guiding a lower limb atherosclerosis patient The theoretical section of this thesis consists of atherosclerosis, the factors that affect its genesis, a section discussing the treatment of the illness and also the significance of guidance, interaction and commitment to the treatment in patient guidance. Atherosclerosis (also known as arteriosclerotic vascular disease or ASVD) is arteriosclerosis and occurs in both larger and minor arterial blood vessels, such as the aorta. In ASVD, the section of the aorta that leads to the lower body or its branch becomes congested. Lower limb ASVD can lead to ischemia. Factors that affect the genesis of lower limb ASVD are: smoking, diabetes, elevated blood sugar and disturbance in blood flow or circulation. Information based on professionalism is often questioned nowadays. The furtherance of health that takes place between the patient and a healthcare professional is based on interaction. Committing to the treatment refers to the client actively and responsibly taking actions as required by the disease while cooperating and interacting with the healthcare personnel. The purpose of our thesis was to help nurses develop their work with guiding lower limb ASVD patients, and it was executed as a qualitative research. The material was gathered by interviewing four nurses that actively work with lower limb ASVD patients, and it consisted of 3 different themes: guidance, interaction and committing to the treatment. In this thesis we used content-based information retrieval method (n=4) to which the basis was the original expressions of the interviewed nurses. The material was analyzed and sorted after the theme-centered interview frame. In the results we found how important it was to consider risk factors in the guidance. The greatest risk factor appeared to be smoking. The patient’s age had a major effect on the interaction situation, and the guidance of the elderly patients was considered challenging at times. The interviewed nurses felt that the biggest challenge in patient guidance was lack of time. The nurses should be given more time to make sure the patients receive adequate guidance. The purpose of this thesis is to give nurses new information for guidance and interaction with the patients and also further theoretical information of the guidance, interaction and commitment to the treatment. The complete thesis was delivered to Meilahti hospital’s department of surgery where nurses work with lower limb ASVD patients