Cutaneous and mucosal human papillomaviruses differ in net surface charge, potential impact on tropism

Papillomaviruses can roughly be divided into two tropism groups, those infecting the skin, including the genus beta PVs, and those infecting the mucosa, predominantly genus alpha PVs. The L1 capsid protein determines the phylogenetic separation between beta types and alpha types and the L1 protein is most probably responsible for the first interaction with the cell surface. Virus entry is a known determinant for tissue tropism and to study if interactions of the viral capsid with the cell surface could affect HPV tropism, the net surface charge of the HPV L1 capsid proteins was analyzed and HPV-16 (alpha) and HPV-5 (beta) with a mucosal and cutaneous tropism respectively were used to study heparin inhibition of uptake. The negatively charged L1 proteins were all found among HPVs with cutaneous tropism from the beta- and gamma-PV genus, while all alpha HPVs were positively charged at pH 7.4. The linear sequence of the HPV-5 L1 capsid protein had a predicted isoelectric point (pI) of 6.59 and a charge of -2.74 at pH 7.4, while HPV-16 had a pI of 7.95 with a charge of +2.98, suggesting no interaction between HPV-5 and the highly negative charged heparin. Furthermore, 3D-modelling indicated that HPV-5 L1 exposed more negatively charged amino acids than HPV-16. Uptake of HPV-5 (beta) and HPV-16 (alpha) was studied in vitro by using a pseudovirus (PsV) assay. Uptake of HPV-5 PsV was not inhibited by heparin in C33A cells and only minor inhibition was detected in HaCaT cells. HPV-16 PsV uptake was significantly more inhibited by heparin in both cells and completely blocked in C33A cells

Kanta-asiakkaiden tyytyväisyys kanta-asiakasviestintään. Dropin uutiskirjeiden sisällöt, kanavavalinta ja ajoitus.

Tutkimuksessani tarkasteltiin Dropin-nimisen katumuoti- ja lautailulajien monikanava- myymälän kanta-asiakkaiden tyytyväisyyttä sähköpostitse lähetettäviin uutiskirjeisiin. Tavoitteena oli analysoida kanta-asiakkaiden kokemuksia tiedonsaannista, kanta-asia- kasviestinnän sisällöstä, kanavavalinnasta ja viestinnän ajoituksesta. Analyysin lähtö- kohtana oli kanta-asiakasviestit ja niiden sisältö. Tutkimuksen aineistona oli kanta-asi- akkaille lähetetty kysely, johon vastasi 1350 asiakaista. Sovelsin viestintätyytyväi- syyden tutkimukseen löyhästi vajeteoriaa, joka on kehitetty sisäisen viestinnän tutkimi- seen. Tutkielman teoreettisena viitekehyksenä käytin sidosryhmäanalyysia. Tutkimuksessa oli kaksi tutkimusosaa. Ensimmäisessä suoritin kanta-asiakasviesteille sisällönanalyysin. Toisessa osassa hyödynsin sisällönanalyysin avulla muodostamiani kategorioita rakentaessani kyselylomakkeen. Kyselyssä vastaajat arvioivat kanta-asia- kasviestien sisältöjen kiinnostavuutta, vaikutusta ostopäätösten syntyyn sekä viestien sisällöistä haluamaansa ja saamaansa tietoa. Lisäksi vastaajat kertoivat mielipiteensä viestinnän ajoitukseen ja kanavalintaan. Analysoin kyselytutkimuksen vastauksia kvan- titatiivisesti tarkastelemalla niiden kokonaismääriä ja prosentteja. Tutkimuksessa selvisi, että kanta-asiakkaat lukevat uutiskirjeitä aktiivisesti ja ovat tyy- tyväisiä viestinnän kanavavalintaan ja melko tyytyväisiä viestinnän sisältöön. Sisältöi- hin ei liittynyt suuria vajeita. Tärkeimmiksi viestinnän sisällöiksi koettiin tarjoukset, kanta-asiakkaan edut, Dropinin järjestämät kilpailut ja arvonnat sekä uusimmat tuotteet. Vajeita koettiin jonkin verran kaikkien sisältöjen suhteen. Naisten ja miesten välillä ei tiedonsaannissa ilmennyt suuria eroja. Viestinnän sisällöistä vähiten tietoa saatiin tuotteiden teknisistä ominaisuuksista, tuot- teiden vaihto- ja palautusoikeuksista sekä lajivälineistä. Tutkimuksessa selvisi myös, että valittu kanava on kanta-asiakkaille mieluisin. Ajoitukseen yrityksen tulisi kiinnittää huomiota, sillä suurin osa kanta-asiakkaista toivoi yhteydenpidon olevan tiheämpää.fi=Opinnäytetyö kokotekstinä PDF-muodossa.|en=Thesis fulltext in PDF format.|sv=Lärdomsprov tillgängligt som fulltext i PDF-format

Hyvinvoiva yrittäjä – hyvinvoiva yritys

Opas tarjoaa helposti toteutettavia ideoita ja ajatuksia yrittäjille oman hyvinvoinnin ja työolojen kohentamiseksi.1

Yrittäjäyhdistys yrittäjän työhyvinvointia edistämässä

Elän täydesti! -teema antaa ajatuksia ja ideoita, miten yrittäjäyhdistys voi tukea jäsenistönsä hyvinvointia.Hanke: Pienetkin pinnalle – mikroyritykset hyvinvointiin Varsinais-Suomessa 2009–2012.1

Maintenance of respiratory chain function in mouse hearts with severely impaired mtDNA transcription

The basal mitochondrial transcription machinery is essential for biogenesis of the respiratory chain and consists of mitochondrial RNA polymerase, mitochondrial transcription factor A (TFAM) and mitochondrial transcription factor B2. This triad of proteins is sufficient and necessary for mtDNA transcription initiation. Abolished mtDNA transcription caused by tissue-specific knockout of TFAM in the mouse heart leads to early onset of a severe mitochondrial cardiomyopathy with lethality within the first post-natal weeks. Here, we describe a mouse model expressing human TFAM instead of the endogenous mouse TFAM in heart. These rescue mice have severe reduction in mtDNA transcription initiation, but, surprisingly, are healthy at the age of 52 weeks with near-normal steady-state levels of transcripts. In addition, we demonstrate that heavy-strand mtDNA transcription normally terminates at the termination-associated sequence in the control region. This termination is abolished in rescue animals resulting in heavy (H)-strand transcription of the entire control region. In conclusion, we demonstrate here the existence of an unexpected mtDNA transcript stabilization mechanism that almost completely compensates for the severely reduced transcription initiation in rescue hearts. Future elucidation of the underlying molecular mechanism may provide a novel pathway to treat mitochondrial dysfunction in human pathology

Molecular crowding defines a common origin for the Warburg effect in proliferating cells and the lactate threshold in muscle physiology

Aerobic glycolysis is a seemingly wasteful mode of ATP production that is seen both in rapidly proliferating mammalian cells and highly active contracting muscles, but whether there is a common origin for its presence in these widely different systems is unknown. To study this issue, here we develop a model of human central metabolism that incorporates a solvent capacity constraint of metabolic enzymes and mitochondria, accounting for their occupied volume densities, while assuming glucose and/or fatty acid utilization. The model demonstrates that activation of aerobic glycolysis is favored above a threshold metabolic rate in both rapidly proliferating cells and heavily contracting muscles, because it provides higher ATP yield per volume density than mitochondrial oxidative phosphorylation. In the case of muscle physiology, the model also predicts that before the lactate switch, fatty acid oxidation increases, reaches a maximum, and then decreases to zero with concomitant increase in glucose utilization, in agreement with the empirical evidence. These results are further corroborated by a larger scale model, including biosynthesis of major cell biomass components. The larger scale model also predicts that in proliferating cells the lactate switch is accompanied by activation of glutaminolysis, another distinctive feature of the Warburg effect. In conclusion, intracellular molecular crowding is a fundamental constraint for cell metabolism in both rapidly proliferating- and non-proliferating cells with high metabolic demand. Addition of this constraint to metabolic flux balance models can explain several observations of mammalian cell metabolism under steady state conditions

Respiratory chain complex III deficiency due to mutated BCS1L : a novel phenotype with encephalomyopathy, partially phenocopied in a Bcs1l mutant mouse model

Background: Mitochondrial diseases due to defective respiratory chain complex III (CIII) are relatively uncommon. The assembly of the eleven-subunit CIII is completed by the insertion of the Rieske iron-sulfur protein, a process for which BCS1L protein is indispensable. Mutations in the BCS1L gene constitute the most common diagnosed cause of CIII deficiency, and the phenotypic spectrum arising from mutations in this gene is wide. Results: A case of CIII deficiency was investigated in depth to assess respiratory chain function and assembly, and brain, skeletal muscle and liver histology. Exome sequencing was performed to search for the causative mutation(s). The patient's platelets and muscle mitochondria showed respiration defects and defective assembly of CIII was detected in fibroblast mitochondria. The patient was compound heterozygous for two novel mutations in BCS1L, c.306A > T and c.399delA. In the cerebral cortex a specific pattern of astrogliosis and widespread loss of microglia was observed. Further analysis showed loss of Kupffer cells in the liver. These changes were not found in infants suffering from GRACILE syndrome, the most severe BCS1L-related disorder causing early postnatal mortality, but were partially corroborated in a knock-in mouse model of BCS1L deficiency. Conclusions: We describe two novel compound heterozygous mutations in BCS1L causing CIII deficiency. The pathogenicity of one of the mutations was unexpected and points to the importance of combining next generation sequencing with a biochemical approach when investigating these patients. We further show novel manifestations in brain, skeletal muscle and liver, including abnormality in specialized resident macrophages (microglia and Kupffer cells). These novel phenotypes forward our understanding of CIII deficiencies caused by BCS1L mutations.Peer reviewe

p38γ and p38δ as biomarkers in the interplay of colon cancer and inflammatory bowel diseases

descripción no proporcionada por scopusThis research was funded by the MCIN/AEI/10.13039/501100011033 (PID2019-108349RB100 and SAF2016-79792R) to AC and JJSE; Villum Foundation, grant no. 13152 to KA; by Agencia Estatal de Investigación (PID2019-104867RBI00/AEI/10.13039/501100011033) and the Instituto de Salud Carlos III- Fondo Europeo de Desarrollo Regional (CIBERONC/CB16/12/00273 and ICI20/00057) to AM and AB. PF received MCIN FPI fellowship (BES-2017-080139)

Sensory Ataxic Neuropathy in Golden Retriever Dogs Is Caused by a Deletion in the Mitochondrial tRNATyr Gene

Sensory ataxic neuropathy (SAN) is a recently identified neurological disorder in golden retrievers. Pedigree analysis revealed that all affected dogs belong to one maternal lineage, and a statistical analysis showed that the disorder has a mitochondrial origin. A one base pair deletion in the mitochondrial tRNATyr gene was identified at position 5304 in affected dogs after re-sequencing the complete mitochondrial genome of seven individuals. The deletion was not found among dogs representing 18 different breeds or in six wolves, ruling out this as a common polymorphism. The mutation could be traced back to a common ancestor of all affected dogs that lived in the 1970s. We used a quantitative oligonucleotide ligation assay to establish the degree of heteroplasmy in blood and tissue samples from affected dogs and controls. Affected dogs and their first to fourth degree relatives had 0–11% wild-type (wt) sequence, while more distant relatives ranged between 5% and 60% wt sequence and all unrelated golden retrievers had 100% wt sequence. Northern blot analysis showed that tRNATyr had a 10-fold lower steady-state level in affected dogs compared with controls. Four out of five affected dogs showed decreases in mitochondrial ATP production rates and respiratory chain enzyme activities together with morphological alterations in muscle tissue, resembling the changes reported in human mitochondrial pathology. Altogether, these results provide conclusive evidence that the deletion in the mitochondrial tRNATyr gene is the causative mutation for SAN