Unsatisfactory gene transfer into bone-resorbing osteoclasts with liposomal transfection systems

BACKGROUND: Bone-resorbing osteoclasts are multinucleated cells that are formed via fusion of their hematopoietic stem cells. Many of the details of osteoclast formation, activation and motility remain unsolved. Therefore, there is an interest among bone biologists to transfect the terminally differentiated osteoclasts and follow their responses to the transgenes in vitro. Severe difficulties in transfecting the large, adherent osteoclasts have been encountered, however, making the use of modern cell biology tools in osteoclast research challenging. Transfection of mature osteoclasts by non-viral gene transfer systems has not been reported. RESULTS: We have systematically screened the usefulness of several commercial DNA transfection systems in human osteoclasts and their mononuclear precursor cell cultures, and compared transfection efficacy to adenoviral DNA transfection. None of the liposome-based or endosome disruption-inducing systems could induce EGFP-actin expression in terminally differentiated osteoclasts. Instead, a massive cell death by apoptosis was found with all concentrations and liposome/DNA-ratios tested. Best transfection efficiencies were obtained by adenoviral gene delivery. Marginal DNA transfection was obtained by just adding the DNA to the cell culture medium. When bone marrow-derived CD34-positive precursor cells were transfected, some GFP-expression was found at the latest 24 h after transfection. Large numbers of apoptotic cells were found and those cells that remained alive, failed to form osteoclasts when cultured in the presence of RANKL and M-CSF, key regulators of osteoclast formation. In comparison, adenoviral gene delivery resulted in the transfection of CD34-positive cells that remained GFP-positive for up to 5 days and allowed osteoclast formation. CONCLUSION: Osteoclasts and their precursors are sensitive to liposomal transfection systems, which induce osteoclast apoptosis. Gene transfer to mononuclear osteoclast precursors or differentiated osteoclasts was not possible with any of the commercial transfection systems tested. Osteoclasts are non-dividing, adherent cells that are difficult to grow as confluent cultures, which may explain problems with transfection reagents. Large numbers of α(v)β(3 )integrin on the osteoclast surface allows adenovirus endocytosis and infection proceeds in dividing and non-dividing cells efficiently. Viral gene delivery is therefore currently the method of choice for osteoclast transfection

Turvemaiden digitaalinen kartoitus ja turvepeltolohkojen tunnistaminen

Maatalouden turvemaiden ilmasto- ja vesistöpäästöjen vähentäminen edellyttää turvepeltolohkojen tunnistamista, mutta maaperätieto ei ole ollut riittävän tarkkaa tähän tarkoitukseen. Raportissa esitellyn työn tavoitteena oli tuottaa tarkennettua paikkatietoa turvemaiden esiintymisestä ja paksuudesta turvepeltolohkojen tunnistamiseksi. Uusi paikkatietoaineisto turvemaiden esiintymisestä ja paksuudesta luotiin hyödyntämällä koneoppimismallinnusta. Mallinnus tehtiin Random Forest -menetelmällä. Turpeen esiintymistä selittäviksi aineistoiksi valmisteltiin 117 kpl koko maan kattavia satelliitti- ja lentoalustoilta mitattuja kaukokartoitusaineistoja ja geologista paikkatietoaineistoa. Koneoppimismallin opettamista ja testausta varten koottiin 3,5 miljoonaa maaperähavaintoa, josta 70 % käytettiin mallin opetukseen ja 30 % mallin riippumattomaan testaukseen. Mallinnuksessa ennustettiin turvepaksuusluokkien ≥ 10 cm, ≥ 30 cm, ≥ 40 cm ja > 60 cm esiintymistä 50 m × 50 m rasteriresoluutiossa ja ennusteet tuotettiin maankäyttömuodosta riippumatta kaikille maa-alueille. Malliennusteiden tarkkuus oli korkea. Turvepaksuusluokat pystyttiin erottelemaan muista maalajeista ja turvepaksuusluokista 89–96 % tarkkuudella. Tarkkuudet olivat korkeimmillaan ohuissa turvepaksuusluokissa ja hieman heikompia paksuissa luokissa. Maatalousmailla vähintään 30 cm paksun turvemaan alaksi arvoitiin 273 000 ha, mikä on noin 11 % maatalousmaa-alasta. Tästä pinta-alasta 73 % turvekerros oli > 60 cm. Saamamme arvio maatalousmaiden turvemaiden (≥ 30 cm) pinta-alasta on 8 600 ha suurempi kuin mitä mittakaavaltaan 1:200 000 maaperäkartasta voidaan arvioida. Peltolohkokohtainen tarkastelu osoitti, että turve-ennusteet mahdollistavat turvealan ja -paksuuden arvioimisen yksittäisillä peltolohkoilla. Esimerkiksi turvepeltolohkot, joilla on vähintään 50 % alastaan ≥30 cm paksu turvekerros, tunnistettiin yli 90 % tarkkuudella. Uusi paikkatietoaineisto Turpeen paksuus 1.0/2023 tarkentaa aikaisempaa tietoa turvemaiden esiintymisestä ja paksuudesta koko maassa. Aineiston luokittelutarkkuus ja alueellinen erottelukyky ovat olemassa olevia maaperäkartta-aineistoja parempia ja sen avulla tunnistetaan aikaisemmin kartoittamattomia turvemaita. Yleistarkkuusmetriikat raportoidaan jokaiselle luokittelulle erikseen ja epävarmuuksien hajautuminen on esitetty Random Forest -puiden yksimielisyyden avulla rasterisolukohtaisesti. Uudet turve-ennusteet tuovat uusia mahdollisuuksia maaperään ja maankäyttöön liittyvien toimintojen suunnittelun, ohjaukseen ja vaikutusten arviointiin, sekä tutkimukseen

История развития физической культуры и спорта на Урале в дореволюционный период

На сегодняшний день становится чрезвычайно актуальным рассмотрение феномена физической культуры и спорта сквозь призму принципа историзма. Существует еще много неизвестного в истории физической культуры, что требует переоценки событий, фактов с позиции современност

Job strain and chronic obstructive pulmonary disease exacerbations: an individual-participant meta-analysis.

To the Editor:Chronic obstructive pulmonary disease (COPD) is a major cause of mortality and disability worldwide (1). The clinical course of COPD is characterised by exacerbations, which can be minor and manageable at home or in primary care, or severe, leading to hospitalisation or even death. Known causes of exacerbations include tobacco smoke, air pollution, dusts and fumes, and respiratory infections (1, 2). One less well understood risk factor is stress, which could plausibly lead to COPD exacerbations as it can trigger inflammation (3, 4) and is associated with increased smoking (5), which are both implicated in COPD pathology (2). Work is an important source of stress in the age groups in which COPD is typically diagnosed (1, 6). However, we are not aware of previous investigations of work-related stress and the risk of COPD exacerbations.In this study, we examined the associations between job strain (the most widely studied conceptualisation of work-related stress) and severe COPD exacerbations using individual-level data from 10 prospective cohort studies from the Individual Participant Data Meta-analysis in Working Populations (IPD-Work) Consortium (7). Job strain is defined as a combination of high demands (excessive amounts of work) and low control (having little influence on what tasks to

Job strain and tobacco smoking: an individual-participant data meta-analysis of 166,130 adults in 15 European studies.

Tobacco smoking is a major contributor to the public health burden and healthcare costs worldwide, but the determinants of smoking behaviours are poorly understood. We conducted a large individual-participant meta-analysis to examine the extent to which work-related stress, operationalised as job strain, is associated with tobacco smoking in working adults

Job strain as a risk factor for clinical depression: systematic review and meta-analysis with additional individual participant data

Background. Adverse psychosocial working environments characterized by job strain (the combination of high demands and low control at work) are associated with an increased risk of depressive symptoms among employees, but evidence on clinically diagnosed depression is scarce. We examined job strain as a risk factor for clinical depression.Method. We identified published cohort studies from a systematic literature search in PubMed and PsycNET and obtained 14 cohort studies with unpublished individual-level data from the Individual-Participant-Data Meta-analysis in Working Populations (IPD-Work) Consortium. Summary estimates of the association were obtained using random-effects models. Individual-level data analyses were based on a pre-published study protocol.Results. We included six published studies with a total of 27 461 individuals and 914 incident cases of clinical depression. From unpublished datasets we included 120 221 individuals and 982 first episodes of hospital-treated clinical depression. Job strain was associated with an increased risk of clinical depression in both published [relative risk (RR) = 1.77, 95% confidence interval (CI) 1.47-2.13] and unpublished datasets (RR = 1.27, 95% CI 1.04-1.55). Further individual participant analyses showed a similar association across sociodemographic subgroups and after excluding individuals with baseline somatic disease. The association was unchanged when excluding individuals with baseline depressive symptoms (RR = 1.25, 95% CI 0.94-1.65), but attenuated on adjustment for a continuous depressive symptoms score (RR = 1.03, 95% CI 0.81-1.32).Conclusions. Job strain may precipitate clinical depression among employees. Future intervention studies should test whether job strain is a modifiable risk factor for depression.</p

Simultaneous HPLC analysis of fat-soluble vitamins in selected animal products after small-scale extraction

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