Hsp70 binds reversibly to proteasome inhibitor-induced protein aggregates and evades autophagic clearance in ARPE-19 cells

Age-related macular degeneration (AMD) is characterized primarily by degeneration of the macular retinal pigment epithelium (RPE) that secondarily leads to cell death of photoreceptors and impaired central vision. Hallmarks of AMD are accumulation of lysosomal lipofuscin and extracellular drusen, which indicate impaired proteolysis in RPE cells. Cellular proteostasis is strongly regulated by molecular chaperones such as Hsp70 and proteasomal and autophagic clearance systems. We have recently shown that autophagy receptor SQSTM1/p62 binds irreversibly to proteasome inhibitor–induced perinuclear protein aggregates and undergoes autophagic clearance in RPE cell cultures. Revealing decreased autophagy, SQSTM1/p62 accumulates in macular area of donor AMD patient samples. In this study, we show that Hsp70 binds reversibly to proteasome inhibitor–induced perinuclear protein aggregates and does not become degraded by autophagy in ARPE-19 cells. Our observation reveals new opportunities to use a cytoprotective Hsp70 as a therapy target in the prevention of RPE cell degeneration and development of AMD

Developing fine-grained nationwide predictions of valuable forests using biodiversity indicator bird species

Publisher Copyright: © 2021 The Authors. Ecological Applications published by Wiley Periodicals LLC on behalf of The Ecological Society of America.The use of indicator species in forest conservation and management planning can facilitate enhanced preservation of biodiversity from the negative effects of forestry and other uses of land. However, this requires detailed and spatially comprehensive knowledge of the habitat preferences and distributions of selected focal indicator species. Unfortunately, due to limited resources for field surveys, only a small proportion of the occurrences of focal species is usually known. This shortcoming can be circumvented by using modelling techniques to predict the spatial distribution of suitable sites for the target species. Airborne laser scanning (ALS) and other remote sensing (RS) techniques have the potential to provide useful environmental data covering systematically large areas for these purposes. Here, we focused on six bird of prey and woodpecker species known to be good indicators of boreal forest biodiversity values. We used known nest sites of the six indicator species based on nestling ringing records. Thus, the most suitable nesting sites of these species provide important information for biodiversity-friendly forest management and conservation planning. We developed fine-grained, i.e., 96 x 96 m grid cell resolution, predictive maps across the whole of Finland of the suitable nesting habitats based on ALS and other RS data and spatial information on the distribution of important forest stands for the six studied biodiversity indicator bird species based on nesting habitat suitability modelling, i.e., the MaxEnt model. Habitat preferences of the study species, as determined by MaxEnt, were in line with the previous knowledge of species-habitat relations. The proportion of suitable habitats of these species in protected areas was considerable, but our analysis also revealed many potentially high-quality forest stands outside protected areas. However, many of these sites are increasingly threatened by logging due to increased pressures for using forests for bioeconomy and forest industry based on National Forest Strategy. Predicting habitat suitability based on information on the nest sites of indicator species provides a new tool for systematic conservation planning over large areas in boreal forests in Europe, and corresponding approach would also be feasible and recommendable elsewhere where similar data are available.The use of indicator species in forest conservation and management planning can facilitate enhanced preservation of biodiversity from the negative effects of forestry and other uses of land. However, this requires detailed and spatially comprehensive knowledge of the habitat preferences and distributions of selected focal indicator species. Unfortunately, due to limited resources for field surveys, only a small proportion of the occurrences of focal species is usually known. This shortcoming can be circumvented by using modeling techniques to predict the spatial distribution of suitable sites for the target species. Airborne laser scanning (ALS) and other remote sensing (RS) techniques have the potential to provide useful environmental data covering systematically large areas for these purposes. Here, we focused on six bird of prey and woodpecker species known to be good indicators of boreal forest biodiversity values. We used known nest sites of the six indicator species based on nestling ringing records. Thus, the most suitable nesting sites of these species provide important information for biodiversity-friendly forest management and conservation planning. We developed fine-grained, that is, 96 x 96 m grid cell resolution, predictive maps across the whole of Finland of the suitable nesting habitats based on ALS and other RS data and spatial information on the distribution of important forest stands for the six studied biodiversity indicator bird species based on nesting-habitat suitability modeling, that is, the MaxEnt model. Habitat preferences of the study species, as determined by MaxEnt, were in line with the previous knowledge of species-habitat relations. The proportion of suitable habitats of these species in protected areas (PAs) was considerable, but our analysis also revealed many potentially high-quality forest stands outside PAs. However, many of these sites are increasingly threatened by logging because of increased pressures for using forests for bioeconomy and forest industry based on National Forest Strategy. Predicting habitat suitability based on information on the nest sites of indicator species provides a new tool for systematic conservation planning over large areas in boreal forests in Europe, and a corresponding approach would also be feasible and recommendable elsewhere where similar data are available.Peer reviewe

Expression of VEGFA-regulating miRNAs and mortality in wet AMD

MicroRNAs (miRNAs) regulate gene expression; many of them act in the retinal pigment epithelium (RPE), and RPE degeneration is known to be a critical factor in age-related macular degeneration (AMD). Repeated injections with anti-VEGFA (vascular endothelial growth factor A) are the only effective therapy in wet AMD. We investigated the correlation between the expression of 18 miRNAs involved in the regulation of the VEGFA gene in serum of 76 wet AMD patients and 70 controls. Efficacy of anti-VEGFA treatment was evaluated by counting the number of injections delivered up to 12 years. In addition, we compared the relative numbers of deaths in patient with AMD and control groups. We observed a decreased expression of miR-34-5p, miR-126-3p, miR-145-5p and miR-205-5p in wet AMD patients as compared with controls. These miRNAs are involved in the regulation of angiogenesis, cytoprotection and protein clearance. No miRNA was significantly correlated with the treatment outcome. Wet AMD patients had greater mortality than controls, and their survival was inversely associated with the number of anti-VEGFA injections per year. No association was observed between miRNA expression and mortality. Our study emphasizes the need to clarify the role of miRNA regulation in AMD pathogenesis.Peer reviewe

Developing a spatially explicit modelling and evaluation framework for integrated carbon sequestration and biodiversity conservation: application in southern Finland

The challenges posed by climate change and biodiversity loss are deeply interconnected. Successful co-managing of these tangled drivers requires innovative methods that can prioritize and target management actions against multiple criteria, while also enabling cost-effective land use planning and impact scenario assessment. This paper synthesises the development and application of an integrated multidisciplinary modelling and evaluation framework for carbon and biodiversity in forest systems. By analysing and spatio-temporally modelling carbon processes and biodiversity elements, we determine an optimal solution for their co-management in the study landscape. We also describe how advanced Earth Observation measurements can be used to enhance mapping and monitoring of biodiversity and ecosystem processes. The scenarios used for the dynamic models were based on official Finnish policy goals for forest management and climate change mitigation. The development and testing of the system were executed in a large region in southern Finland (Kokemäenjoki basin, 27 024 km2) containing highly instrumented LTER (Long-Term Ecosystem Research) stations; these LTER data sources were complemented by fieldwork, remote sensing and national data bases. In the study area, estimated total net emissions were currently 4.2 TgCO2eq a-1, but modelling of forestry measures and anthropogenic emission reductions demonstrated that it would be possible to achieve the stated policy goal of carbon neutrality by low forest harvest intensity. We show how this policy-relevant information can be further utilised for optimal allocation of set-aside forest areas for nature conservation, which would significantly contribute to preserving both biodiversity and carbon values in the region. Biodiversity gain in the area could be increased without a loss of carbon-related benefits.The challenges posed by climate change and biodiversity loss are deeply interconnected. Successful co-managing of these tangled drivers requires innovative methods that can prioritize and target management actions against multiple criteria, while also enabling cost-effective land use planning and impact scenario assessment. This paper synthesises the development and application of an integrated multidisciplinary modelling and evaluation framework for carbon and biodiversity in forest systems. By analysing and spatio-temporally modelling carbon processes and biodiversity elements, we determine an optimal solution for their co-management in the study landscape. We also describe how advanced Earth Observation measurements can be used to enhance mapping and monitoring of biodiversity and ecosystem processes. The scenarios used for the dynamic models were based on official Finnish policy goals for forest management and climate change mitigation. The development and testing of the system were executed in a large region in southern Finland (Kokemäenjoki basin, 27,024 km2) containing highly instrumented LTER (Long-Term Ecosystem Research) stations; these LTER data sources were complemented by fieldwork, remote sensing and national data bases. In the study area, estimated total net emissions were currently 4.2 TgCO2eq a−1, but modelling of forestry measures and anthropogenic emission reductions demonstrated that it would be possible to achieve the stated policy goal of carbon neutrality by low forest harvest intensity. We show how this policy-relevant information can be further utilized for optimal allocation of set-aside forest areas for nature conservation, which would significantly contribute to preserving both biodiversity and carbon values in the region. Biodiversity gain in the area could be increased without a loss of carbon-related benefits.Peer reviewe

Loss of NRF-2 and PGC-1α genes leads to retinal pigment epithelium damage resembling dry age-related macular degeneration

Age-related macular degeneration (AMD) is a multi-factorial disease that is the leading cause of irreversible and severe vision loss in the developed countries. It has been suggested that the pathogenesis of dry AMD involves impaired protein degradation in retinal pigment epithelial cells (RPE). RPE cells are constantly exposed to oxidative stress that may lead to the accumulation of damaged cellular proteins, DNA and lipids and evoke tissue deterioration during the aging process. The ubiquitin-proteasome pathway and the lysosomal/autophagosomal pathway are the two major proteolytic systems in eukaryotic cells. NRF-2 (nuclear factor-erythroid 2-related factor-2) and PGC-1 alpha (peroxisome proliferator-activated receptor gamma coactivator-1 alpha) are master transcription factors in the regulation of cellular detoxification. We investigated the role of NRF-2 and PGC-1 alpha in the regulation of RPE cell structure and function by using global double knockout (dKO) mice. The NRF-2/PGC-1 alpha dKO mice exhibited significant age-dependent RPE degeneration, accumulation of the oxidative stress marker, 4-HNE (4-hydroxynonenal), the endoplasmic reticulum stress markers GRP78 (glucose-regulated protein 78) and ATF4 (activating transcription factor 4), and damaged mitochondria. Moreover, levels of protein ubiquitination and autophagy markers p62/SQSTM1 (sequestosome 1), Beclin-1 and LC3B (microtubule associated protein 1 light chain 3 beta) were significantly increased together with the Iba-1 (ionized calcium binding adaptor molecule 1) mononuclear phagocyte marker and an enlargement of RPE size. These histopathological changes of RPE were accompanied by photoreceptor dysmorphology and vision loss as revealed by electroretinography. Consequently, these novel findings suggest that the NRF-2/PGC-1 alpha dKO mouse is a valuable model for investigating the role of proteasomal and autophagy clearance in the RPE and in the development of dry AMD.Peer reviewe

Listaamattoman pk-yrityksen arvonmääritys satakuntalaisessa tilitoimistossa

Tämä pro gradu -tutkielma käsittelee yrityksen arvonmääritys prosessia ja sen eri menetelmiä. Tutkielman tarkoituksena on suorittaa arvonmääritysprosessi kokonaisuudessaan listaamattomalle kirjapitoalan pk-yritykselle, hyödyntäen vapaata kassavirtamallia sekä lisäarvomallia. Tutkielmassa arvonmääritysprosessi on jaettu viiteen eri vaiheeseen, jotka ovat strateginen analyysi, tilinpäätösanalyysi, tulevan kehityksen arviointi ja yrityksen arvon määrittäminen valittuja arvonmääritysmenetelmiä hyödyntäen. Näiden eri osa-alueiden tarkoituksena on muodostaa selkeä käsitys kohdeyrityksen menneisyydestä, tästä hetkestä ja tulevasta. Vapaa kassavirtamalli sekä lisäarvomalli ovat hyvin yleisiä arvonmäärittämisen työkaluja ja sopivat hyvin tutkielman kohteena olevan listaamattoman pk-yrityksen arvon määrittämiseen. Tutkielmassa analysoidaan valittujen mallien toimivuutta kohdeyrityksen tapauksessa, tarkastellaan arvonmäärityksen ongelmakohtia ja pohditaan arvonmäärityksen merkitystä osana pk-yrityksen tulevaisuutta. Tämä tutkielma noudattaa toiminta-analyyttiselle tutkimusotteelle tyypillisiä piirteitä, eli tarkoituksena on kuvata teorian ja empirian avulla yksittäistä ilmiötä mahdollisimman tarkasta. Tämän tutkielman tapauksessa satakuntalaista tilitoimistoa. Tutkielman tavoitteena on selvittää, mikä merkitys arvonmäärityksellä on osana pk-yrityksen tulevaisuutta, sekä mikä on kohdeyrityksen arvo valittuja menetelmiä hyödyntäen? Kohdeyrityksen taloudellista tilannetta ja tulevan kehityksen ennakointia on arvioitu strategisen analyysin ja tilinpäätös analyysin pohjalta. Kohdeyrityksen riskikäsitys on muodostettu kokonaispääoman tuottovaatimusta hyödyntäen. Viimeisessä vaiheessa eli kohdeyrityksen arvonmäärityksessä, yrityksen arvoksi muodostui vapaalla kassavirta malilla noin 2,4 miljoonaa euroa ja lisäarvomallia hyödyntäen noin 1,1 miljoonaa euroa. Valittujen mallien tulokset antavat toisistaan selkeästi poikkeavat tulokset. Nämä poikkeamat arvioidaan tässä tutkielmassa johtuvan vapaan kassavirtamallin suuremmasta herkkyydestä eri muuttujille. Vapaan kassavirtamallin herkkyys kokonaispääoman tuottovaatimuksen ja kasvuennusteiden muutoksille oli selvästi suurempi kuin lisäarvonmallissa. Vaikka tässä tutkielmassa lisäarvomallin koetaan antavan realistisempi tulos yrityksen arvosta, ei sitä tule pitää absoluuttisena totuutena. Yrityksen arvon tarkastelu perustuu pitkälti tulevaisuuteen ja näin ollen yksiselitteisesti oikeaa arvoa on mahdoton määrittää. Aina lopussa yrityksen arvo määräytyy sen eri osapuolten välisenä tulkintana yrityksen tilanteesta ja erilaiset menetelmät antavat vain alustan tulkinnan toteutukselle. Arvonmääritys antaa paljon arvokasta tietoa ja sitä tulisikin hyödytään pk-yrityksissä enemmän osana liiketoiminnan kehittämistä

Absence of collagen XVIII in mice causes age-related insufficiency in retinal pigment epithelium proteostasis

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Absence of collagen XVIII in mice causes age-related insufficiency in retinal pigment epithelium proteostasis

Collagen XVIII has the structural propertiesof both collagen and proteoglycan. It has beenfound at the basement membrane/stromal interfacewhere it is thought to mediate their attachment.Endostatin, a proteolytic fragment from collagenXVIII C-terminal end has been reported to possessanti-angiogenic properties. Age-related vision loss incollagen XVIII mutant mice has been accompaniedwith a pathological accumulation of deposits under theretinal pigment epithelium (RPE). We have recentlydemonstrated that impaired proteasomal andautophagy clearance are associated with the pathogenesisof age-related macular degeneration. Thisstudy examined the staining levels of proteasomal andautophagy markers in the RPE of different ages of theCol18a1-/- mice. Eyes from 3, 6–7, 10–13 and18 months old mice were enucleated and embedded inparaffin according to the routine protocol. Sequential5 lm-thick parasagittal samples were immunostainedfor proteasome and autophagy markers ubiquitin (ub),SQSTM1/p62 and beclin-1. The levels of immunopositivityin the RPE cells were evaluated by confocalmicroscopy. Collagen XVIII knock-out mice hadundergone age-related RPE degeneratio