| 1. |
- Gram, Magnus, et al.
(författare)
-
Increased levels of hemoglobin and alpha1-microglobulin in Huntington's disease.
- 2012
-
Ingår i: Frontiers in Bioscience (Elite Edition). - 1945-0508. ; 4, s. 950-957
-
Tidskriftsartikel (refereegranskat)abstract
- Hemoglobin released from damaged erythrocytes is a major pro-oxidant, generator of free radicals and inflammatory mediator. Huntington's disease is an inherited neurodegenerative disorder characterized by both neurological and systemic abnormalities, in which oxidative stress has been suggested as a possible pathogenic mechanism. In the present work we have investigated levels of hemoglobin and markers of oxidative damage, including the heme- and radical-scavenger alpha1-microglobulin, in plasma and urine samples from two separate sample cohorts, including controls, premanifest gene carriers and subjects at different stages of Huntington's disease. The results show statistically significant increased levels of hemoglobin and alpha1-microglobulin in Huntington's disease urine samples. Interestingly, urine hemoglobin levels correlate with clinical severity. The results suggest that hemolysis may be linked to the pathogenesis of Huntington's disease and that assay of hemoglobin and alpha1-microglobulin may provide biomarkers that are linked to biologically relevant processes.
|
|
| 2. |
- Hinkula, Jorma, et al.
(författare)
-
Immunization with DNA Plasmids Coding for Crimean-Congo Hemorrhagic Fever Virus Capsid and Envelope Proteins and/or Virus-Like Particles Induces Protection and Survival in Challenged Mice
- 2017
-
Ingår i: Journal of Virology. - : AMER SOC MICROBIOLOGY. - 0022-538X .- 1098-5514. ; 91:10
-
Tidskriftsartikel (refereegranskat)abstract
- Crimean-Congo hemorrhagic fever virus (CCHFV) is a bunyavirus causing severe hemorrhagic fever disease in humans, with high mortality rates. The requirement of a high-containment laboratory and the lack of an animal model hampered the study of the immune response and protection of vaccine candidates. Using the recently developed interferon alpha receptor knockout (IFNAR(- / -)) mouse model, which replicates human disease, we investigated the immunogenicity and protection of two novel CCHFV vaccine candidates: a DNA vaccine encoding a ubiquitin-linked version of CCHFV Gc, Gn, and N and one using transcriptionally competent virus-like particles (tc-VLPs). In contrast to most studies that focus on neutralizing antibodies, we measured both humoral and cellular immune responses. We demonstrated a clear and 100% efficient preventive immunity against lethal CCHFV challenge with the DNA vaccine. Interestingly, there was no correlation with the neutralizing antibody titers alone, which were higher in the tc-VLP-vaccinated mice. However, the animals with a lower neutralizing titer, but a dominant cell-mediated Th1 response and a balanced Th2 response, resisted the CCHFV challenge. Moreover, we found that in challenged mice with a Th1 response (immunized by DNA/DNA and boosted by tc-VLPs), the immune response changed to Th2 at day 9 postchallenge. In addition, we were able to identify new linear B-cell epitope regions that are highly conserved between CCHFV strains. Altogether, our results suggest that a predominantly Th1-type immune response provides the most efficient protective immunity against CCHFV challenge. However, we cannot exclude the importance of the neutralizing antibodies as the surviving immunized mice exhibited substantial amounts of them. IMPORTANCE Crimean-Congo hemorrhagic fever virus (CCHFV) is responsible for hemorrhagic diseases in humans, with a high mortality rate. There is no FDAapproved vaccine, and there are still gaps in our knowledge of the immune responses to infection. The recently developed mouse models mimic human CCHF disease and are useful to study the immunogenicity and the protection by vaccine candidates. Our study shows that mice vaccinated with a specific DNA vaccine were fully protected. Importantly, we show that neutralizing antibodies are not sufficient for protection against CCHFV challenge but that an extra Th1-specific cellular response is required. Moreover, we describe the identification of five conserved B-cell epitopes, of which only one was previously known, that could be of great importance for the development of diagnostics tools and the improvement of vaccine candidates.
|
|
| 3. |
- Kristiansson, Amanda, et al.
(författare)
-
α1-Microglobulin (A1M) Protects Human Proximal Tubule Epithelial Cells from Heme-Induced Damage In Vitro
- 2020
-
Ingår i: International Journal of Molecular Sciences. - : MDPI AG. - 1661-6596 .- 1422-0067. ; 21:16
-
Tidskriftsartikel (refereegranskat)abstract
- Oxidative stress is associated with many renal disorders, both acute and chronic, and has also been described to contribute to the disease progression. Therefore, oxidative stress is a potential therapeutic target. The human antioxidant α1-microglobulin (A1M) is a plasma and tissue protein with heme-binding, radical-scavenging and reductase activities. A1M can be internalized by cells, localized to the mitochondria and protect mitochondrial function. Due to its small size, A1M is filtered from the blood into the glomeruli, and taken up by the renal tubular epithelial cells. A1M has previously been described to reduce renal damage in animal models of preeclampsia, radiotherapy and rhabdomyolysis, and is proposed as a pharmacological agent for the treatment of kidney damage. In this paper, we examined the in vitro protective effects of recombinant human A1M (rA1M) in human proximal tubule epithelial cells. Moreover, rA1M was found to protect against heme-induced cell-death both in primary cells (RPTEC) and in a cell-line (HK-2). Expression of stress-related genes was upregulated in both cell cultures in response to heme exposure, as measured by qPCR and confirmed with in situ hybridization in HK-2 cells, whereas co-treatment with rA1M counteracted the upregulation. Mitochondrial respiration, analyzed with the Seahorse extracellular flux analyzer, was compromised following exposure to heme, but preserved by co-treatment with rA1M. Finally, heme addition to RPTE cells induced an upregulation of the endogenous cellular expression of A1M, via activation of the nuclear factor erythroid 2-related factor 2 (Nrf2)-pathway. Overall, data suggest that A1M/rA1M protects against stress-induced damage to tubule epithelial cells that, at least partly, can be attributed to maintaining mitochondrial function.
|
|
| 4. |
- Sanchez, Diego, et al.
(författare)
-
Molecular interactions of the neuronal GPI-anchored lipocalin Lazarillo
- 2008
-
Ingår i: Journal of Molecular Recognition. - : Wiley. - 1099-1352 .- 0952-3499. ; 21:5, s. 312-322
-
Tidskriftsartikel (refereegranskat)abstract
- Lazarillo, a glycoprotein involved in axon growth and guidance in the grasshopper embryo, is the only member of the lipocalin family that is attached to the cell surface by a GPI anchor. Recently, the study of Lazarillo homologous genes in Drosophila and mouse has revealed new functions in the regulation of lifespan, stress resistance and neurodegeneration. Here we report an analysis of biochemical properties of Lazarillo to gain insight into the molecular basis of its physiological function. Recombinant forms of the grasshopper protein were expressed in two different systems to test: (1) potential binding of several hydrophobic ligands; (2) protein-protein homophilic interactions; and (3) whether interaction with the function-blocking mAb 10E6 interferes with ligand binding. We tested 10 candidate ligands (retinoic acid, heme, bilirubin, biliverdin, ecdysterone, juvenile hormone, farnesol, arachidonic acid, linoleic acid and palmitic acid), and monitored binding using electrophoretic mobility shift, absorbance spectrum, and fluorimetry assays. Our work indicates binding to heme and retinoic acid, resulting in increased electrophoretic mobility, as well as to fatty acids, resulting in multimerization. Retinoic acid and fatty acids binding were confirmed by fluorescence titration, and heme binding was confirmed with absorbance spectrum assays. We demonstrate that Lazarillo oligomerizes in solution and can form clusters in the plasma membrane when expressed and GPI-anchored to the cell surface, however it is unable to mediate cell-cell adhesion. Finally, by ligand-mAb competition experiments we show that ligand-binding alone cannot be the key factor for Lazarillo to perform its function during axonal growth in the grasshopper embryo. Copyright (C) 2008 John Wiley & Sons, Ltd.
|
|
| 5. |
- Silins, Isabella, 1983-, et al.
(författare)
-
First-in-human evaluation of [18F]CETO : a novel tracer for adrenocortical tumours
- 2023
-
Ingår i: European Journal of Nuclear Medicine and Molecular Imaging. - : Springer Nature. - 1619-7070 .- 1619-7089. ; 50:2, s. 398-409
-
Tidskriftsartikel (refereegranskat)abstract
- Purpose[11C]Metomidate positron emission tomography (PET) is currently used for staging of adrenocortical carcinoma and for lateralization in primary aldosteronism (PA). Due to the short half-life of carbon-11 and a high non-specific liver uptake of [11C]metomidate there is a need for improved adrenal imaging methods. In a previous pre-clinical study para-chloro-2-[18F]fluoroethyletomidate has been proven to be a specific adrenal tracer. The objective is to perform a first evaluation of para-chloro-2-[18F]fluoroethyletomidate positron emission computed tomography ([18F]CETO-PET/CT) in patients with adrenal tumours and healthy volunteers.MethodsFifteen patients underwent [18F]CETO-PET/CT. Five healthy volunteers were recruited for test-retest analysis and three out of the five underwent additional [15O]water PET/CT to measure adrenal blood flow. Arterial blood sampling and tracer metabolite analysis was performed. The kinetics of [18F]CETO were assessed and simplified quantitative methods were validated by comparison to outcome measures of tracer kinetic analysis.ResultsUptake of [18F]CETO was low in the liver and high in adrenals. Initial metabolization was rapid, followed by a plateau. The kinetics of [18F]CETO in healthy adrenals and all adrenal pathologies, except for adrenocortical carcinoma, were best described by an irreversible single-tissue compartment model. Standardized uptake values (SUV) correlated well with the uptake rate constant K1. Both K1 and SUV were highly correlated to adrenal blood flow in healthy controls. Repeatability coefficients of K1, SUV65–70, and SUV120 were 25, 22, and 17%.ConclusionsHigh adrenal uptake combined with a low unspecific liver uptake suggests that 18F]CETO is a suitable tracer for adrenal imaging. Adrenal SUV, based on a whole-body scan at 1 h p.i., correlated well with the net uptake rate Ki.Trial registrationClinicalTrials.gov, NCT05361083 Retrospectively registered 29 April 2022. at, https://clinicaltrials.gov/ct2/show/NCT05361083
|
|
| 6. |
- Åkerström, Sara, 1976, et al.
(författare)
-
Evaluation of individual characteristics in partnering projects
- 2007
-
Ingår i: Proceedings of CIB World congress Cape Town, May 14-17 in Cape Town, South Africa. - 1920017046
-
Konferensbidrag (refereegranskat)abstract
- Partnering arrangements have become more common in the Swedish construction sector over the last years. However, studies show that some issues are not yet established in partnering projects, e.g., the evaluation of “soft parameters” in recruitment processes of staff. Accordingly, there is a need of further investigation of the process of evaluating key persons’ attitudes and cooperation capabilities. Based on a study of three cases, partnering projects, this paper aims at improving the methods that has been used for evaluation of companies’ representatives in partnering projects. Focus is on the relationships between actors, individual characteristics, i.e., soft parameters that may have effect on project outcomes. The finding indicates fundamental problems in the evaluation process of key staff’ characteristics and also deficiencies in the methods used for evaluation purpose. To develop a satisfactory relationship between actors in a partnering project, key persons should fulfil four criteria of competence, integrity, benevolence and attitudes to teamwork. Each of these criteria contains large number of characteristics and attitudes that should be discussed by actors at an early stage of a partnering project.
|
|
| 7. |
- Åkerström, Sara
(författare)
-
SARS coronavirus : the role of accessory proteins and nitric oxide in the replication cycle
- 2008
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Severe acute respiratory syndrome coronavirus (SARS-CoV) caused the first pandemic of the 21st century. The etiological agent was identified as a novel coronavirus. Until then, human coronaviruses (HCoVs) had only been known to cause the common cold . Data indicated that the virus originated from animals. The palm civet (Paguma larvata)was identified as the source of transmitting the virus to humans, however not to be the natural host, but most likely acting as an amplifier before the virus was transmitted to humans. SARS-CoV has 14 potential open reading frames (ORFs). Eight of those are specific for SARS and encodes for the accessory proteins. The accessory proteins of SARS-CoV have no known sequence homology to any other accessory proteins found in coronaviruses. The knowledge about the function of most of the accessory proteins are limited. In addition to the four main structural proteins; the spike (S), the envelope (E), the membrane (M) and the nucleocapsid (N) protein, the accessory protein 3a of SARS-CoV has been shown to be a minor structural protein. SARS-CoV, as the name implies, causes severe lower respiratory syndrome. Person-to- person transmission has been through infectious droplets, and the overall mortality rate is ~10% but can vary with age. Most SARS patients were treated with antiviral drugs and glucocorticoids since no specific treatment has been available. Inhalation of nitric oxide (NO) has been given to only a few patients showed a positive effect. In this thesis we have investigated the importance of accessory proteins 3a/3b and 7a/7b in the replication cycle. Further we compared the neutralizing properties of the endo- and the ectodomian of 3a. We also investigated the antiviral effect of NO on SARS-CoV, and a possible mechanism behind an antiviral effect. Short interfering RNA (siRNA) was designed to specifically target sgRNA 2, 3 and 7 expressing the S, 3a/3b and 7a/7b protein respectively. The yield of progeny virus was significantly reduced for all three siRNAs. The amount of progeny virus was to some extent lower for siRNA 7, which could be due to the fact that siRNA 7 were able to silence both 7a/7b and 8a/8b protein. Cells expressing the siRNAs specifically silenced the expression of targeted proteins without affecting the infection shown by expression of the N protein. The 3a protein was further investigated, comparing neutralizing properties of antibodies towards the endodomain and the ectodomain of 3a. Antibodies towards both ends were able to detect 3a in lysate from infected Vero E6 cells. However, only antibodies against the ectodomain showed neutralizing properties in Vero E6 cells. In order to investigate the antiviral affect of NO on SARS-CoV, both an exogenous NO donor, and endogenously produced NO was used. We showed that NO has a clear antiviral effect on SARS-CoV, inhibiting the replication cycle of the virus. To investigate the mechanism behind the antiviral effect of NO, we first confirmed that NO per se was exerting the observed antiviral effect, and not through peroxynitrite interaction. By using a cell-cell fusion assay, we showed that NO inhibits the fusion step by reducing palmitoylation of the SARS-CoV S protein.
|
|
