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Global age-sex-specific mortality, life expectancy, and population estimates in...

Schumacher, AE; Kyu, HH; Aali, A; Abbafati, C; Abbas, J; Abbasgholizadeh, R; Abbasi, MA; Abbasian, M; Abd ElHafeez, S; Abdelmasseh, M; Abd Elsalam, S

Background: Estimates of demographic metrics are crucial to assess levels and trends of population health outcomes. The profound impact of the COVID-19 pandemic on populations worldwide has underscored the need for timely estimates to understand this unprecedented event within the context of long-term population health trends. The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2021 provides n...


Inter-laboratory variation in measurement of DNA damage by the alkaline comet a...

Moller, P; Azqueta, A; Collia, M; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, S; Scavone, F

The comet assay is a simple and versatile method for measurement of DNA damage in eukaryotic cells. More specifically, the assay detects DNA migration from agarose gel-embedded nucleoids, which depends on assay conditions and the level of DNA damage. Certain steps in the comet assay procedure have substantial impact on the magnitude of DNA migration (e.g. electric potential and time of electrophoresis). Inter-l...


DNA strand break levels in cryopreserved mononuclear blood cell lines measured ...

Moller, P; Azqueta, A; Rodriguez-Garraus, A; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, S

The comet assay is widely used in biomonitoring studies for the analysis of DNA damage in leukocytes and peripheral blood mononuclear cells. Rather than processing blood samples directly, it can be desirable to cryopreserve whole blood or isolated cells for later analysis by the comet assay. However, this creates concern about artificial accumulation of DNA damage during cryopreservation. In this study, 10 labo...


Long-term cryopreservation of potassium bromate positive assay controls for mea...

Moller, P; Azqueta, A; Rodriguez-Garraus, A; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Ristori, S

The formamidopyrimidine DNA glycosylase (Fpg)-modified comet assay is widely used for the measurement of oxidatively generated damage to DNA. However, there has not been a recommended long-term positive control for this version of the comet assay. We have investigated potassium bromate as a positive control for the Fpg-modified comet assay because it generates many Fpg-sensitive sites with a little concurrent g...


Visual comet scoring revisited: a guide to scoring comet assay slides and obtai...

Moller, P; Azqueta, A; Sanz-Serrano, J; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Bastos, VC; Langie, SAS; Jensen, A; Scavone, F

Measurement of DNA migration in the comet assay can be done by image analysis or visual scoring. The latter accounts for 20%-25% of the published comet assay results. Here we assess the intra- and inter-investigator variability in visual scoring of comets. We include three training sets of comet images, which can be used as reference for researchers who wish to use visual scoring of comets. Investigators in 11 ...


A conformational switch controlling the toxicity of the prion protein

Frontzek, K; Bardelli, M; Senatore, A; Henzi, A; Reimann, RR; Bedir, S; Marino, M; Hussain, R; Jurt, S; Meisl, G; Pedotti, M; Mazzola, F; Siligardi, G

Prion infections cause conformational changes of the cellular prion protein (PrPC) and lead to progressive neurological impairment. Here we show that toxic, prion-mimetic ligands induce an intramolecular R208-H140 hydrogen bond (‘H-latch’), altering the flexibility of the α2–α3 and β2–α2 loops of PrPC. Expression of a PrP2Cys mutant mimicking the H-latch was constitutively toxic, whereas a PrPR207A mutant unabl...


AAV Vector-Mediated Antibody Delivery (A-MAD) in the Central Nervous System

Marino, M; Holt, MG

In the last four decades, monoclonal antibodies and their derivatives have emerged as a powerful class of therapeutics, largely due to their exquisite targeting specificity. Several clinical areas, most notably oncology and autoimmune disorders, have seen the successful introduction of monoclonal-based therapeutics. However, their adoption for treatment of Central Nervous System diseases has been comparatively ...


Astrocyte-targeted gene delivery of interleukin 2 specifically increases brain-...

Yshii, L; Pasciuto, E; Bielefeld, P; Mascali, L; Lemaitre, P; Marino, M; Dooley, J; Kouser, L; Verschoren, S; Lagou, V; Kemps, H; Gervois, P; Boer, A

The ability of immune-modulating biologics to prevent and reverse pathology has transformed recent clinical practice. Full utility in the neuroinflammation space, however, requires identification of both effective targets for local immune modulation and a delivery system capable of crossing the blood-brain barrier. The recent identification and characterization of a small population of regulatory T (Treg) cells...


AAV-mediated delivery of an anti-BACE1 VHH alleviates pathology in an Alzheimer...

Marino, M; Zhou, L; Rincon, MY; Callaerts-Vegh, Z; Verhaert, J; Wahis, J; Creemers, E; Yshii, L; Wierda, K; Saito, T; Marneffe, C; Voytyuk, I

Single domain antibodies (VHHs) are potentially disruptive therapeutics, with important biological value for treatment of several diseases, including neurological disorders. However, VHHs have not been widely used in the central nervous system (CNS), largely because of their restricted blood-brain barrier (BBB) penetration. Here, we propose a gene transfer strategy based on BBB-crossing adeno-associated virus (...


Potassium bromate as positive assay control for the Fpg-modified comet assay

Møller, P; Muruzabal, D; Bakuradze, T; Richling, E; Bankoglu, EE; Stopper, H; Langie, SAS; Azqueta, A; Jensen, A; Scavone, F; Giovannelli, L

The comet assay is a popular assay in biomonitoring studies. DNA strand breaks (or unspecific DNA lesions) are measured using the standard comet assay. Oxidative stress-generated DNA lesions can be measured by employing DNA repair enzymes to recognise oxidatively damaged DNA. Unfortunately, there has been a tendency to fail to report results from assay controls (or maybe even not to employ assay controls). We b...


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