| 1. |
- Carlsson, Lena, et al.
(författare)
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High Concentrations of the Angiogenic Peptide VEGF-A in Seminal Fluid and its Association to Prostasomes
- 2016
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Ingår i: Clinical Laboratory. - 1433-6510. ; 62:8, s. 1515-1520
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Tidskriftsartikel (refereegranskat)abstract
- Background: Angiogenesis is the formation of new blood vessels by capillary sprouting from pre-existing vessels. This process is associated with increased expression of angiogenic factors like vascular endothelial growth factor (VEGF). The VEGF family consists of five members denoted VEGF-A, B, C, D and placenta growth factor (PlGF). Prostasomes are exosome-like extracellular vesicles existing in seminal plasma. The present study aimed at investigating the possible relationship between VEGF-A in seminal fluid and blood plasma and the prostasomal association of VEGF-A.Methods: Measurement of VEGF-A concentrations was carried out in seminal plasma from 40 males and in blood plasma from 40 male blood donors utilizing commercial ELISA kits. The prostasomal association of VEGF-A was investigated by flow cytometry.Results: We found highly elevated concentrations of VEGF-A in seminal fluid (median value 150000 pg/mL) compared with those of blood plasma. Flow cytometric analysis showed that VEGF-A is bound to the surface of prostasomes.Conclusions: Prostasomes and seminal plasma contain the angiogenic factor VEGF-A in high concentrations exceeding that of blood plasma by 1000 times.
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| 2. |
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| 3. |
- Dubois, Louise, et al.
(författare)
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Human erythrocyte-derived nanovesicles can readily be loaded with doxorubicin and act as anticancer agents
- 2018
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Ingår i: Cancer Research Frontiers. - : Cancer Research Frontiers. - 2328-5249. ; 4:1, s. 13-26
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Tidskriftsartikel (refereegranskat)abstract
- Purpose: In future therapeutics new formulas are needed that assure lower doses, fewer side effects, targeted administration and protection of the drug from degradation. In a first step to fulfil the requirements defined above, we carried out an in vitro study by developing a new procedure to encapsulate drugs using native vesicles first from prostasomes and then from erythrocyte membranes known to be well tolerated. The new method for production of drug delivery vesicles utilized osmotic loading of detergent resistant membranes (DRMs).Materials and methods: DRMs of prostasomes and prepared human erythrocyte membranes were extracted and separated in a sucrose gradient at a density of 1.10 g/mL containing 1% Triton X-100. These DRMs were characterized by electron microscopy (transmission and scanning EM) and loaded with low and high molecular compounds. PC3 prostate cancer cells were treated with doxorubicin loaded DRMs in triplicate. DAPI (nuclear fluorescent stain) was included and fluorescence microscopic pictures were taken before the cells were trypsinized and counted after 48h.Results: The content of the well separated band was observed ultrastructurally as small spherical, double layered membrane vesicles, (DRM vesicles) which harbored hyperosmolar sucrose of the gradient. Encapsulated hyperosmolar sucrose induced a transient osmotic lysis of the DRM vesicles when suspended in isotonic buffer containing loading molecules allowing vesicular inclusion. After this proof of concept, the method was finally employed for doxorubicin loading of DRM vesicles from human erythrocytes. When incubating such vesicles with PC3 cells a complete arrest of growth was observed in sharp contrast to PC3 cells incubated with plain doxorubicin in similar conditions.Conclusion: The present results open up new possibilities for using DRM vesicles as drug delivery vesicles.
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| 4. |
- Dubois, Louise, et al.
(författare)
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Proteomic profiling of detergent resistant membranes (lipid rafts) of prostasomes
- 2015
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Ingår i: Molecular & Cellular Proteomics. - 1535-9476 .- 1535-9484. ; 14:11, s. 3015-3022
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Tidskriftsartikel (refereegranskat)abstract
- Prostasomes are exosomes derived from prostate epithelial cells through exocytosis by multivesicular bodies. Prostasomes have a bilayered membrane and readily interact with sperm. The membrane lipid composition is unusual with a high contribution of sphingomyelin at the expense of phosphatidylcholine and saturated and monounsaturated fatty acids are dominant. Lipid rafts are liquid-ordered domains that are more tightly packed than the surrounding non-raft phase of the bilayer. Lipid rafts are proposed to be highly dynamic, submicroscopic assemblies that float freely within the liquid disordered membrane bilayer and some proteins preferentially partition into the ordered raft domains. We asked the question whether lipid rafts do exist in prostasomes and, if so, which proteins might be associated with them. Prostasomes of density range 1.13-1.19g/mL were subjected to density gradient ultracentrifugation in sucrose fabricated by phosphate buffered saline (PBS) containing 1% Triton X-100 with capacity for banding at 1.10g/mL, i.e. the classical density of lipid rafts. Prepared prostasomal lipid rafts (by gradient ultracentrifugation) were analyzed by mass spectrometry and electron microscopy. The clearly visible band on top of 1.10g/mL sucrose in the Triton X-100 containing gradient was subjected to LC-MS/MS and more than 370 lipid raft associated proteins were identified. Several of them were involved in intraluminal vesicle formation, e.g. tetraspanins, ESCRTs and Ras-related proteins. This is the first comprehensive LC-MS/MS profiling of proteins in lipid rafts derived from exosomes. Data are available via ProteomeXchange with identifier PXD002163.
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| 5. |
- Inayat, S, et al.
(författare)
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High levels of cathepsins B, L and S in human seminal plasma and their association with prostasomes
- 2012
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Ingår i: Andrologia. - : Hindawi Limited. - 0303-4569 .- 1439-0272. ; 44:6, s. 423-427
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Tidskriftsartikel (refereegranskat)abstract
- Semen is a heterogenous and complex fluid with different functions, some of them well known, others still obscure. The aim of this study was to investigate the presence of cathepsins B, S and L in human seminal plasma and their possible associations with other semen variables. Cathepsin B, L and S concentrations were measured in seminal plasma from 99 men utilising commercial ELISA kits. Seminal plasma cathepsin B was approximately 70 times higher, while the cathepsin L values were approximately 500 times higher and the cathepsin S values approximately 40 times higher in seminal plasma than in a group of serum samples. The study shows that seminal plasma contains high levels of cathepsins B, L and S. All three cathepsins were also bound to the surface of prostasomes.
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| 6. |
- Larssen, Pia, et al.
(författare)
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Tracing Cellular Origin of Human Exosomes Using Multiplex Proximity Extension Assay
- 2017
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Ingår i: Molecular & Cellular Proteomics. - 1535-9476 .- 1535-9484. ; 16:3, s. 502-511
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Tidskriftsartikel (refereegranskat)abstract
- Extracellular vesicles (EVs) are membrane-coated objects such as exosomes and microvesicles, released by many cell-types. Their presence in body fluids and the variable surface composition and content render them attractive potential biomarkers. The ability to determine their cellular origin could greatly move the field forward. We used multiplex proximity extension assays (PEA) to identify with high specificity and sensitivity the protein profiles of exosomes of different origins, including seven cell lines and two different body fluids. By comparing cells and exosomes, we successfully identified the cells originating the exosomes. Furthermore, by principal component analysis of protein patterns human milk EVs and prostasomes released from prostate acinar cells clustered with cell lines from breast and prostate tissues, respectively. Milk exosomes uniquely expressed CXCL5, MIA and KLK6, while prostasomes carried NKX31, GSTP1 and SRC, highlighting that EVs originating from different origins express distinct proteins. In conclusion, PEA provides a powerful protein screening tool in exosome research, for purposes of identifying the cell source of exosomes, or new biomarkers in diseases such as cancer and inflammation.
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| 7. |
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| 8. |
- Nickel, Katrin F., et al.
(författare)
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The polyphosphate-factor XII pathway drives coagulation in prostate cancer-associated thrombosis
- 2015
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Ingår i: Blood. - : American Society of Hematology. - 0006-4971 .- 1528-0020. ; 126:11, s. 1379-1389
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Tidskriftsartikel (refereegranskat)abstract
- Cancer is a leading cause of thrombosis. We identify a new procoagulant mechanism that contributes to thromboembolism in prostate cancer and allows for safe anticoagulation therapy development. Prostate cancer-mediated procoagulant activity was reduced in plasma in the absence of factor XII or its substrate of the intrinsic coagulation pathway factor XI. Prostate cancer cells and secreted prostasomes expose long chain polyphosphate on their surface that colocalized with active factor XII and initiated coagulation in a factor XII-dependent manner. Polyphosphate content correlated with the procoagulant activity of prostasomes. Inherited deficiency in factor XI or XII or high-molecular-weight kininogen, but not plasma kallikrein, protected mice from prostasome-induced lethal pulmonary embolism. Targeting polyphosphate or factor XII conferred resistance to prostate cancer-driven thrombosis in mice, without increasing bleeding. Inhibition of factor XII with recombinant 3F7 antibody reduced the increased prostasome-mediated procoagulant activity in patient plasma. The data illustrate a critical role for polyphosphate/factor XII-triggered coagulation in prostate cancer-associated thrombosis with implications for anticoagulation without therapy-associated bleeding in malignancies.
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| 9. |
- Ronquist, Göran, et al.
(författare)
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Human Prostasomes Contain Chromosomal DNA
- 2009
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Ingår i: The Prostate. - : Wiley. - 0270-4137 .- 1097-0045. ; 69:7, s. 737-743
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND. The aim of this study was to perform a comprehensive evaluation of the occurrence of DNA in human prostasomes. METHODS. Prostasomes were purified from seminal fluid (seminal prostasomes) and from PC-3-cells (PC-3 cell prostasomes). DNA extracted from both sources of prostasomes was visualized on agarose gels. Further, theDNAwas cloned and sequenced (13 clones from seminal prostasomal DNAand 16 clones from PC-3 cell prostasomal DNA) and identified by alignment in the BLAST-nucleotide search database. In order to decide if the DNA was internally or externally located in/on prostasomes, prostasomes were treated with nuclease (DNase) and A260 was measured before and after treatment. Additionally, flow cytometric studies were performed with membrane permeable and membrane impermeable DNA stains. RESULTS. We identified human chromosomal DNA in purified prostasomes from both sources and treatment with DNase demonstrated that the prostasome-shielded DNA was protected from enzyme attack. Membrane-permeable DNA stain raised the fluorescence contrary to membrane-impermeable stain. Clearly discernible nucleic acid of prostasomes was separated on 1% agarose gel yieldingDNAfragments of about 13 kbp and below with a marked band at about 1 kbp. Cloning and sequencing of 13 fragments from seminal prostasomes and 16 from PC-3 cell prostasomes revealed a chromosomal origin of the DNA. In purified seminal prostasomes, 4 out of 13 DNA clones featured gene sequences (31%). The corresponding figure for PC3-derived prostasomes was 4 out of 16 clones featuring gene sequences (25%). CONCLUSION. Human prostasomes contain chromosomal DNA. Both nuclease treatment and differential DNA stainings indicated an inside location of the prostasomal DNA. Our findings suggest a DNA-delivery function of prostasomes to sperm cells.
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| 10. |
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| 11. |
- Ronquist, Göran K, et al.
(författare)
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Biochemical characterization of stallion prostasomes and comparison to their human counterparts
- 2013
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Ingår i: Systems biology in reproductive medicine. - : Informa UK Limited. - 1939-6376 .- 1939-6368. ; 59:6, s. 297-303
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Tidskriftsartikel (refereegranskat)abstract
- Release of nanometer-sized prostasomes into human and equine semen suggests essential functions in their relationships with sperm cells and the fertilization process. The two types of prostasomes displayed ultrastructural similarities, albeit the human prostasomes were somewhat larger than the stallion prostasomes. A high ratio of saturated fatty acids was characteristic for the two prostasome types. Electrophoretic separation systems revealed an equine prostasomal pattern different from that of human. The 21 distinctive low molecular weight protein spots in the 2D-gel (with no counterparts in human prostasomes) were identified via peptide mass fingerprinting, several of which may be different isoforms. Out of the three high molecular weight bands characteristic for human prostasomes (CD10, CD13, and CD26), CD10 and CD13 were retrieved in equine prostasomes. We present some new proteins of horse prostasomes not found in their human counterparts. Further studies are warranted to reveal the function of these proteins.
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| 12. |
- Ronquist, Göran K., et al.
(författare)
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Prostasomal DNA Characterization and Transfer Into Human Sperm
- 2011
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Ingår i: Molecular Reproduction and Development. - : Wiley. - 1040-452X .- 1098-2795. ; 78:7, s. 467-476
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Tidskriftsartikel (refereegranskat)abstract
- Human prostasomes, exosome-like microvesicles secreted by acinar cells of the prostate gland, contain chromosomal DNA. Agarose gel electrophoresis of DNA from seminal prostasomes displayed fragments of over 12 kb and smaller, with a distinct band around 1 kb that was excised, cloned, and sequenced. The sequences showed 8 out of 25 clones (32%) originating from genes. We elaborated the concept further by carrying out a genome-wide DNA copy number analysis of prostasomal DNA, hypothesizing that human prostasomes contain fragments of DNA randomly selected from the entire genome. Acridine orange-stained prostasomes were incubated with freshly prepared sperm for different times, and a transfer of acridine orange-stained prostasomal DNA to sperm (preferentially the head region) was observed. Fluorescence microscopy of slices in the center of 14 optical slides of the sperm head displayed an even fluorescence rather than a halo-like one, indicating DNA-uptake rather than just binding along the sperm head membrane.
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| 13. |
- Ronquist, Göran K, et al.
(författare)
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Prostasomes are heterogeneous regarding size and appearance but affiliated to one DNA-containing exosome family
- 2012
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Ingår i: The Prostate. - : Wiley. - 0270-4137 .- 1097-0045. ; 72:16, s. 1736-1745
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND:Prostate acinar epithelial cells release microvesicles (prostasomes) that possess pleiotropic biological effects relevant for successful fertilization. Prostasomes are formed in a similar way as exosomes but are heterogeneous as regards size and appearance. Like exosomes they are thought to be mediators of intercellular communication.METHODS:We prepared seminal prostasomes in accordance with the prevailing protocol for exosome preparation including passage through a 0.2 µm filter and centrifugation in a sucrose gradient.RESULTS:We compared the "filterable prostasomes" with those trapped on the filter ("nonfilterable prostasomes") and, qualitatively, no conspicuous differences were apparent regarding ultrastructure and SDS-PAGE banding pattern. Moreover, both types of prostasomes contained DNA fragments and Western blot revealed presence of prostate specific membrane antigen (PSMA), CD38, and annexin A1.CONCLUSIONS: Reasonably, prostasomes could be included in the exosome family and be regarded as one entity containing chromosomal DNA.
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| 14. |
- Ronquist, Göran, et al.
(författare)
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Prostasome-derived proteins capable of eliciting an immune response in prostate cancer patients
- 2006
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Ingår i: International Journal of Cancer. - : Wiley. - 0020-7136 .- 1097-0215. ; 119:4, s. 847-853
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Tidskriftsartikel (refereegranskat)abstract
- Prostate cancer consistently remains a difficult clinical enigma. Therefore, the development of novel strategies for diagnosis and treatment (e.g. immunotherapy) of prostate cancer is essential. We tried to identify the prostasome-derived proteins that were immunogenic in prostate cancer patients. Prostate cancer patients’ sera (n 5 44) with high enzyme-linked immunosorbent assay (ELISA) titers against prostasomes were selected for immunoblotting against purified seminal prostasomes. The SDS-PAGE and immunoblotting experiments were performed with Bio-Rad systems. Twenty-five of the recognized proteins were isolated and analyzed by means of mass spectrometry. Out of 44 patients’ sera, 31 (70%) demonstrated in immunoblotting experiments reactivity against several prostasomal protein bands in the molecular weight range of 10– 200 kDa. Some of the bands (55, 70 and 170 kDa) were more frequently recognized by the patients’ sera. Concomitantly run control sera generated only very weak or no bands at all. The most frequently occurring prostasomal proteins were identified as heat shock proteins (HSP 70, 71) and clusterin. This study identified the most important molecular targets of autoantibodies against prostasomes generated in connection with the development of prostate cancer in man. These immunogenic prostasomal proteins could be appropriate target molecules for specific immunotherapy of prostate cancer patients.
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| 15. |
- Ronquist, Göran, et al.
(författare)
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Proteomic analysis of prostate cancer metastasis : derived prostasomes
- 2010
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Ingår i: Anticancer Research. - 0250-7005 .- 1791-7530. ; 30:2, s. 285-290
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Tidskriftsartikel (refereegranskat)abstract
- The secretory epithelial cells of the prostate gland use sophisticated vehicles in the form of prostasomes to relay important information to sperm cells in semen. This prostasome-forming and secretory ability of the epithelial cells is also preserved in poorly differentiated prostate cancer cells. We investigated prostasomes from vertebral metastases of prostate cancer, taken from the operating field at surgery, directly taken care of under protease inhibitory conditions for later 2-dimensional electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) protein characterization. A total of 104 spots were punched out for identification. Twenty five unique protein spots had a MALDI-TOF above 49 and another 5 proteins were determined by MS/MS. The remaining 74 spots were either identical to already determined proteins or had no reliable score. Annexins A1, A3, and A5 as well as dimethylarginine dimethylaminohydrolase 1 were among the identified proteins. The annexins and dimethylarginine dimethylaminohydrolase 1 found in cancer-derived prostasomes can act, among other things, as angiogenic factors and can increase the vascular development in the neighborhood of the tumor. Cancer-derived prostasomes may play an important role in the interaction between tumor cells and their environment.
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| 16. |
- Ronquist, Göran, et al.
(författare)
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Serum antibodies against prostasomal clusterin in prostate cancer patients
- 2008
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Ingår i: Scandinavian Journal of Clinical and Laboratory Investigation. - : Informa UK Limited. - 0036-5513 .- 1502-7686. ; 68:3, s. 219-227
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Tidskriftsartikel (refereegranskat)abstract
- Objective. Clusterin is a ubiquitous secretory sulphated glycoprotein present in prostasomes. It is an antiapoptotic mediator in prostate cancer and is among the most frequently occurring prostasomal proteins immunogenic in prostate cancer patients. The aim of the present study was to investigate the occurrence of anticlusterin antibodies in the serum of patients with prostate cancer and whether there is a relationship between anticlusterin antibody titres and other clinico-pathological variables. Material and methods. Serum samples were collected from 391 consecutive patients with suspected prostate cancer (150 benign prostate and 241 prostate cancer). The patients’ serum samples were used in an ELISA where microtitre wells were coated with purified clusterin from serum of a healthy volunteer. Flow cytometric studies of clusterin and prostasomes were performed. Results. Flow cytometric analyses revealed the presence of clusterin on the surface of seminal prostasomes. Anti-clusterin ELISA titres in sera of patients did not differ significantly from those of a control group. A significant ‘‘inverse’’ correlation existed between anti-clusterin ELISA titres and lymph node metastases (p50.047), but only 11 out of 161 patients had metastases. These titres correlated significantly with total prostate (p50.021) and transitional zone (p50.015) volumes of the patients. Conclusions. The correlation between serum anti-clusterin antibody titres and other clinico-pathological variables was generally weak in prostate cancer patients, although clusterin has been assigned an important role in tumourigenesis and progression of prostate cancer. However, the anti-clusterin antibody titre appeared to be related to prostate volume, correlating to both transitional zone volume and total volume of the prostate.
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| 17. |
- Ronquist, Karl Göran, et al.
(författare)
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Energy-requiring uptake of prostasomes and PC3 cell-derived exosomes into non-malignant and malignant cells
- 2016
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Ingår i: Journal of Extracellular Vesicles. - : Wiley. - 2001-3078. ; 5
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Tidskriftsartikel (refereegranskat)abstract
- Epithelial cells lining the prostate acini release, in a regulated manner (exocytosis), nanosized vesicles called prostasomes that belong to the exosome family. Prostate cancer cells have preserved this ability to generate and export exosomes to the extracellular space. We previously demonstrated that human prostasomes have an ATP-forming capacity. In this study, we compared the capacity of extracellular vesicles (EVs) to generate ATP between normal seminal prostasomes and exosomes secreted by PC3 cells (PC3 exosomes), a prostate cancer cell line. Proteomic analyses identified enzymes of the glycolytic chain in both prostasomes and PC3 exosomes, and we found that both of them were capable of generating ATP when supplied with substrates. Notably, the net production of extracellular ATP was low for prostasomes due to a high ATPase activity contrary to an elevated net ATP level for PC3 exosomes because of their low ATPase activity. The uptake of the 2 types of EVs by normal prostate epithelial cells (CRL2221) and prostate cancer cells (PC3) was visualized and measured, demonstrating differential kinetics. Interestingly, this uptake was dependent upon an ongoing glycolytic flux involving extracellular ATP formation by EVs and/or intracellular ATP produced from the recipient cells. We conclude that the internalization of EVs into recipient cells is an energy-requiring process also demanding an active V-ATPase and the capacity of EVs to generate extracellular ATP may play a role in this process.
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| 18. |
- Ronquist, K Göran, et al.
(författare)
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Human prostasomes express glycolytic enzymes with capacity for ATP production
- 2013
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Ingår i: American Journal of Physiology. Endocrinology and Metabolism. - : American Physiological Society. - 0193-1849 .- 1522-1555. ; 304:6, s. E576-E582
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Tidskriftsartikel (refereegranskat)abstract
- Prostasomes are prostate-derived, exosome-like microvesicles that transmit signaling complexes between the acinar epithelial cells of the prostate and sperm cells. A vast majority of prostasomes has a diameter of 30 - 200 nm and they are generally surrounded by a classical membrane bilayer. Using a selected proteomic approach, it became increasingly clear that prostasomes harbor distinct subsets of proteins that may be linked to adenosine triphosphate (ATP) metabolic turnover that in turn might be of importance in the role of prostasomes as auxiliary instruments in the fertilization process. Among the 21 proteins identified most of the enzymes of anaerobic glycolysis were represented and three of the glycolytic enzymes present are among the ten top proteins found in most exosomes, once again linking prostasomes to the exosome family. Other prostasomal enzymes involved in ATP turnover were adenylate kinase, ATPase, 5'-nucleotidase and hexose transporters. The identified enzymes in their prostasomal context were operational for ATP formation when supplied with substrates. The net ATP production was low due to a high prostasomal ATPase activity that could be partially inhibited by vanadate that was utilized in order to profile the ATP forming ability of prostasomes. Glucose and fructose were equivalent as glycolytic substrates for prostasomal ATP formation and the enzymes involved were apparently surface-located on prostasomes, since an alternative substrate not being membrane-permeable (glyceraldehyde 3-phosphate) was operative, too. There is no clear cut function linked to this subset of prostasomal proteins but some possible roles are discussed.
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| 19. |
- Ronquist, K Göran, et al.
(författare)
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Prostasomes from four different species are able to produce extracellular adenosine triphosphate (ATP)
- 2013
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Ingår i: Biochimica et Biophysica Acta - General Subjects. - : Elsevier BV. - 0304-4165 .- 1872-8006. ; 1830:10, s. 4604-4610
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND:Prostasomes are extracellular vesicles. Intracellularly they are enclosed by another larger vesicle, a so called "storage vesicle" equivalent to a multivesicular body of late endosomal origin. Prostasomes in their extracellular context are thought to play a crucial role in fertilization.METHODS:Prostasomes were purified according to a well worked-out schedule from seminal plasmas obtained from human, canine, equine and bovine species. The various prostasomes were subjected to SDS-PAGE separation and protein banding patterns were compared. To gain knowledge of the prostasomal protein systems pertaining to prostasomes of four different species proteins were analyzed using a proteomic approach. An in vitro assay was employed to demonstrate ATP formation by prostasomes of different species.RESULTS:The SDS-PAGE banding pattern of prostasomes from the four species revealed a richly faceted picture with most protein bands within the molecular weight range of 10-150kDa. Some protein bands seemed to be concordant among species although differently expressed and the number of protein bands of dog prostasomes seemed to be distinctly fewer. Special emphasis was put on proteins involved in energy metabolic turnover. Prostasomes from all four species were able to form extracellular adenosine triphosphate (ATP). ATP formation was balanced by ATPase activity linked to the four types of prostasomes.CONCLUSION:These potencies of a possession of functional ATP-forming enzymes by different prostasome types should be regarded against the knowledge of ATP having a profound effect on cell responses and now explicitly on the success of the sperm cell to fertilize the ovum.GENERAL SIGNIFICANCE:This study unravels energy metabolic relationships of prostasomes from four different species.
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| 20. |
- Ahlström, Katarina, 1966, et al.
(författare)
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Exogenous carbon monoxide does not affect cell membrane energy availability assessed by sarcolemmal calcium fluxes during myocardial ischaemia-reperfusion in the pig
- 2011
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Ingår i: European Journal of Anaesthesiology. - 0265-0215 .- 1365-2346. ; 28:5, s. 356-362
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Tidskriftsartikel (refereegranskat)abstract
- Carbon monoxide is thought to be cytoprotective and may hold therapeutic promise for mitigating ischaemic injury. The purpose of this study was to test low-dose carbon monoxide for protective effects in a porcine model of acute myocardial ischaemia and reperfusion. In acute open-thorax experiments in anaesthetised pigs, pretreatment with low-dose carbon monoxide (5% increase in carboxyhaemoglobin) was conducted for 120 min before localised ischaemia (45 min) and reperfusion (60 min) was performed using a coronary snare. Metabolic and injury markers were collected by microdialysis sampling in the ventricular wall. Recovery of radio-marked calcium delivered locally by microperfusate was measured to assess carbon monoxide treatment effects during ischaemia/reperfusion on the intracellular calcium pool. Coronary occlusion and ischaemia/reperfusion were analysed for 16 animals (eight in each group). Changes in glucose, lactate and pyruvate from the ischaemic area were observed during ischaemia and reperfusion interventions, though there was no difference between carbon monoxide-treated and control groups during ischaemia or reperfusion. Similar results were observed for glycerol and microdialysate Ca-45(2+) recovery. These findings show that a relatively low and clinically relevant dose of carbon monoxide did not seem to provide acute protection as indicated by metabolic, energy-related and injury markers in a porcine myocardial ischaemia/reperfusion experimental model. We conclude that protective effects of carbon monoxide related to ischaemia/reperfusion either require higher doses of carbon monoxide or occur later after reperfusion than the immediate time frame studied here. More study is needed to characterise the mechanism and time frame of carbon monoxide-related cytoprotection.
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| 21. |
- Ahlström, Katarina, 1966, et al.
(författare)
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Metabolic responses in ischemic myocardium after inhalation of carbon monoxide.
- 2009
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Ingår i: Acta Anaesthesiol Scand. - : Wiley. - 1399-6576 .- 0001-5172. ; 53:8, s. 1036-42
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: To clarify the mechanisms of carbon monoxide (CO) tissue-protective effects, we studied energy metabolism in an animal model of acute coronary occlusion and pre-treatment with CO. METHODS: In anesthetized pigs, a coronary snare and microdialysis probes were placed. CO (carboxyhemoglobin 5%) was inhaled for 200 min in test animals, followed by 40 min of coronary occlusion. Microdialysate was analyzed for lactate and glucose, and myocardial tissue samples were analyzed for adenosine tri-phosphate, adenosine di-phosphate, and adenosine mono-phosphate. RESULTS: Lactate during coronary occlusion was approximately half as high in CO pre-treated animals and glucose levels decreased to a much lesser degree during ischemia. Energy charge was no different between groups. CONCLUSIONS: CO in the low-doses tested in this model results in a more favorable energy metabolic condition in that glycolysis is decreased in spite of maintained energy charge. Further work is warranted to clarify the possible mechanistic role of energy metabolism for CO protection.
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| 22. |
- Dubois, Louise, et al.
(författare)
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Malignant Cell-Derived Extracellular Vesicles Express Different Chromogranin Epitopes Compared to Prostasomes
- 2015
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Ingår i: The Prostate. - : Wiley. - 0270-4137 .- 1097-0045. ; 75:10, s. 1063-1073
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND. Prostasomes are nanosized extracellular vesicles exocytosed by prostate epithelial cells. They have been assigned many roles propitious to sperm in favor of fertilization. Prostatic cancer cells can also produce and secrete extracellular vesicles. METHODS. We assessed using ELISA, the surface expression of chromogranin proproteins on prostasomes and malignant extracellular vesicles of four different prostate cancer cell-lines, two hormone sensitive and two hormone refractory. We used a panel of chromogranin A and chromogranin B antibodies against peptides in-between hypothetical cleavage sites along the proproteins. RESULTS. A diverging pattern of chromogranin peptides was apparent when comparing prostasomes and malignant extracellular vesicles indicating a phenotypical change. We also compared western blot patterns (prostasomes and malignant extracellular vesicles) for selected antibodies that displayed high absorbances in the ELISA. Western blot analyses revealed various cleavage patterns of those proproteins that were analyzed in prostasomes and extracellular vesicles. CONCLUSION. Chromogranins are constituents of not only prostasomes but also of malignant prostate cell-derived extracellular vesicles with different amino acid sequences exposed at the membrane surface giving rise to a mosaic pattern. These findings may be of relevance for designing new assays for detection or even possible treatment of prostate cancers.
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| 23. |
- Forshage, Mattias, 1968-, et al.
(författare)
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Quasimodoana, a new Holarctic genus of eucoiline wasps (Hymenoptera, Cynipoidea, Figitidae), with a phylogenetic analysis of related genera
- 2008
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Ingår i: Systematic Entomology. - : Wiley. - 0307-6970 .- 1365-3113. ; 33:2, s. 301-318
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Tidskriftsartikel (refereegranskat)abstract
- A new Holarctic genus, Quasimodoana gen.n. is described for the Palearctic eucoiline wasp Eucoila decipiens Förster, 1869, as Quasimodoana decipiens comb.n. A lectotype is designated for this species. A new North American species is described here as Quasimodoana gibba sp.n. As the new genus has similarities with several eucoiline genera, belonging to two related but distinct lineages, a phylogenetic analysis was carried out, based on 83 morphological characters. The two lineages included in the analysis are referred to here under the tribal names of Eucoilini and Trichoplastini stat.nov. We discuss reasons for adopting available tribal names (rather than 'informal genus groups'), and Trichoplastini is removed from synonymy with Eucoilini. The phylogenetic analysis places Quasimodoana unambiguously within Eucoilini, as a sistergroup to the Trybliographa complex. However, there is only weak support for the monophyly of Eucoilini and for the hypothesized sistergroup relationship between Eucoilini and Trichoplastini.
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| 24. |
- Forshage, Mattias, 1968-
(författare)
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Systematics of Eucoilini : Exploring the diversity of a poorly known group of Cynipoid parasitic wasps
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Many animal species are still undiscovered, even in Sweden. This thesis deals with the wasps of the subfamily Eucoilinae (of Figitidae, Cynipoidea), a poorly known group of small parasitoids of Dipteran flies. In this group, old classifications are chaotic, knowledge of old taxa is scant, phylogenetic work has recently started, and many new taxa remain to be described. In this thesis, an identification key is given to the European genera of Eucolinae, and a tribal classification of the subfamily is reintroduced, based on recent phylogenetic results. Most of the thesis focuses on the tribe Eucoilini. That tribe is analyzed phylogenetically in two analyses, one based on a large morphological dataset, the other on two molecular markers, ribosomal 28S and mitochondrial Cytochrome B. The monophyletic core group of Eucoilini is the Eucoila/Trybliographa complex. It is treated in a big taxonomic revision, based on studies of several hundred possibly relevant type specimens and several thousands of other specimens, and on the phylogenetic analyses. More than 500 names of possible members of this group are investigated, and many new combinations are proposed, of species found not to belong within the group, as well as those that did, including some which are transferred to other genera in accordance with the new phylogenetic results. The new genus Quasimodoana is erected, and 20 new species described (8 from Northern Europe, 12 from elsewhere). More than 180 new combinations, more than 50 new species-level synonyms and ca 10 genus-level synonyms, are proposed. Within this complex, the basal split is found to have been between a Paleotropic, specis-poor lineage (Bothrochacis), and a species-rich and largely Holarctic lineage, dominated by Trybliographa but also including the small genera Eucoila and Linoeucoila. The three are not satisfactorily resolved in the analysis, but still maintained as separate genera for pragmatic reasons. It is hypothesized that this group originated in Asia during the Eocene or Oligocene, attacking calyptrate flies in herbivore dung. Then it separated in a tropical branch retaining the ancestral life history mode, and a Holarctic branch, where some lineages specialised in Anthomyiidae and colonised the microhabitats of fungi and plants, speciating extensively in the boreal forests.
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| 25. |
- Larsson, Anders, et al.
(författare)
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Life style intervention in moderately overweight individuals is associated with decreased levels of cathepsins L and S in plasma
- 2014
-
Ingår i: Annals of Clinical and Laboratory Science. - : The Association of Clinical Scientists. - 0091-7370 .- 1550-8080. ; 44:3, s. 283-285
-
Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Adipose tissue cells produce cathepsins L and S, which have proatherogenic effects. Obesity is strongly linked to atherogenesis, cardiovascular morbidity, and mortality. OBJECTIVE: The aim of the present study was to see if life style interventions/weight reduction could decrease cathepsin L and S levels in blood plasma. METHOD: Study subjects (n=31) were recruited to a life style intervention program aiming at increased physical activity, more healthy eating habits, and weight reduction for most of the participants. Blood samples were collected at inclusion and after 4 and 8 weeks. RESULTS: Cathepsin L was significantly reduced at 4 weeks (p<0.0001) and 8 weeks (p=0.0004). A similar reduction was also seen for cathepsin S at 4 weeks (p=0.03) and 8 weeks (p=0.008). No significant change in fractalkine values was observed at 4 weeks (p=0.58), but a significant increase was apparent at 8 weeks (p=0.0002). CONCLUSION: The intervention program resulted in significant reductions of cathepsin L and S levels in plasma after 4 and 8 weeks of intervention.
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| 26. |
- Larsson, Anders, et al.
(författare)
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Lifestyle intervention is associated with decreased concentrations of circulating pentraxin 3 independent of CRP decrease
- 2013
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Ingår i: Upsala Journal of Medical Sciences. - : Uppsala Medical Society. - 0300-9734 .- 2000-1967. ; 118:3, s. 165-168
-
Tidskriftsartikel (refereegranskat)abstract
- Objectives. Pentraxin 3 (PTX3) is an acute phase marker, which is produced at the site of infection or inflammation in contrast to CRP that is mainly synthesized by the liver. The aim of the present study was to see if lifestyle interventions/weight loss would lead to decreased blood plasma concentrations of PTX3. Methods. Study subjects (n = 31) were recruited to a lifestyle intervention program aiming at increased physical activity, improved eating habits, and weight loss. High-sensitivity C-reactive protein (CRP) and PTX3 methods were used for analysis of CRP and PTX3 in plasma samples collected at inclusion and after 4 and 8 weeks of treatment. Results. Wilcoxon paired samples test showed a significant decrease in PTX3 concentrations from 2068 pg/mL at start to 2007 pg/mL at 4 weeks (P = 0.002) and 1748 pg/mL at 8 weeks (P = 0.003). The PTX3 decrease was not significantly correlated with a corresponding decrease in CRP or weight reduction. Conclusions. The lifestyle intervention program resulted in a significant reduction of circulating concentrations of pentraxin 3 already after 4 and 8 weeks of treatment.
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| 27. |
- Liu, Zhiwei, et al.
(författare)
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The cynipoid genus Paramblynotus : Revision, phylogeny, and historical biogeography (Hymenoptera : Liopteridae)
- 2007
-
Ingår i: Bulletin of the American Museum of Natural History. - 0003-0090 .- 1937-3546. ; :304, s. 3-151
-
Tidskriftsartikel (refereegranskat)abstract
- The genus Paramblynotus is the most species-rich genus of the so-called macrocynipoids, the large cynipoid parasitoids of wood-boring and cone-boring insect larvae. The species range in size from some of the largest to the smallest macrocynipoids, comparable in size to microcynipoids. Paramblynotus members occur on all continents except Europe and Australia, with most species being tropical or subtropical. The biology is poorly known but a few observations indicate that the species are parasitoids of beetle larvae. In this monographic revision of the genus, we present a species-level cladistic analysis based on qualitative and quantitative features of the external morphology. For analysis of quantitative features, we present for the first time a novel coding method, the method of Finite Mixture Coding (FMC) based on k-means clustering, or FMCK. The new method is similar to the FMC method proposed by previous authors in that they both generate codes (character states) for phylogenetic analysis as the direct output of a statistical procedure, thus avoiding the subdivision of quantitative data into discrete states on the basis of arbitrary criteria as with other coding methods. Through incorporating finite mixture analysis and likelihood estimation as used in FMC and k-mean cluster analysis for a priori statistical modeling of component distributions, FMCK is advantageous over FMC in that it can be implemented using readily available statistic programs with k-mean cluster analysis, such as STATISTICA, MINITAB or SYSTAT, available on both PC and Macintosh platforms. We were able to identify 8 quantitative characters among 23 as useful for cladistic analysis by using the new coding method. In total, our character matrix has 132 coded characters. The phylogenetic analysis indicates that species of the previously recognized genus Decelleu form a monophyletic group deeply nested within Paramblynotus. Decellea is therefore synonymized with Paramblynotus, which is separated into seven monophyletic species groups: the virginianus, scaber, ynngambicolars, nigricornis, apeosus, ruficollis, and punclulalus groups. Based on the phylogeny, we reconstruct the historical biogeography of the liopterid subfamily Mayrellinae, consisting of the genera Paramblynotus and Kiefferiellu, using dispersal-vicariance analysis in combination with palaeoenvironmental data. The results suggest that the subfamily originated in the Northern Hemisphere and then expanded its distribution early by way of the Bering area. The divergence between Puramblynotus and Kiefferiellu was apparently associated with the formation of the Rocky Mountains about 50 million years ago. An early Paramblynotus lineage dispersed to Africa from the eastern Palearctic by way of Arabia, and it subsequently diversified along with montane forests in Africa. The relatively high diversity of Paramblynotus in Southeast Asia is considered to be partly caused by the frequent sea level changes since late Oligocene (29 Ma), which drastically changed the land configuration of this area. We end this paper with a taxonomic revision of the genus Paramblynotus, with a total of 92 species treated, including 72 described as new and 20 previously known, of which 18 are redescribed. Keys to the species groups as defined in this paper and to all known species of each species group are provided. For all species, the available information on their biology and distribution are summarized.
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| 28. |
- Löf, Liza, et al.
(författare)
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Detecting individual extracellular vesicles using a multicolor in situ proximity ligation assay with flow cytometric readout
- 2016
-
Ingår i: Scientific Reports. - : Springer Science and Business Media LLC. - 2045-2322. ; 6
-
Tidskriftsartikel (refereegranskat)abstract
- Flow cytometry is a powerful method for quantitative and qualitative analysis of individual cells. However, flow cytometric analysis of extracellular vesicles (EVs), and the proteins present on their surfaces has been hampered by the small size of the EVs - in particular for the smallest EVs, which can be as little as 40 nm in diameter, the limited number of antigens present, and their low refractive index. We addressed these limitations for detection and characterization of EV by flow cytometry through the use of multiplex and multicolor in situ proximity ligation assays (in situ PLA), allowing each detected EV to be easily recorded over background noise using a conventional flow cytometer. By targeting sets of proteins on the surface that are specific for distinct classes of EVs, the method allows for selective recognition of populations of EVs in samples containing more than one type of EVs. The method presented herein opens up for analyses of EVs using flow cytometry for their characterization and quantification.
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| 29. |
- Olovsson, Matts, 1958-, et al.
(författare)
-
Human uterine myocytes retain their energy charge with No gross alterations in morphology for at least 8 days when cultured under anaerobic conditions
- 2000
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Ingår i: Gynecologic and Obstetric Investigation. - : S. Karger AG. - 0378-7346 .- 1423-002X. ; 49:3, s. 165-169
-
Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE:To investigate the ability of human uterine myocytes to grow under anaerobic conditions for a prolonged time period.METHODS:Cells were isolated from fundal myometrium and cultured until subconfluency. The cell type was confirmed by immunostaining for the smooth muscle cell-specific cytoskeletal proteins alpha-actin and desmin. Some cells were further cultured under aerobic conditions and others under anaerobic conditions. Cells were harvested after 0, 4 and 8 days in culture and analyzed for their content of adenylates.RESULTS:Immunostaining revealed that all three preparations contained almost only smooth muscle cells. Energy charge of the myocytes was 0.88 on average at the beginning of the culture experiment. A moderate decrease was noted on day 4 for myocytes grown under both aerobic and anaerobic conditions and no further decrease was noted between days 4 and 8. Morphologically the cells retained their normal appearance and they seemed healthy for at least 8 days in culture under both aerobic and anaerobic conditions.CONCLUSIONS:The results of this study suggest that human myometrial cells can survive for an extended period of time under in vitro conditions regardless of oxygen availability for energy metabolism. This means that anaerobic energy metabolism is enough to sustain vital processes during that period of time.
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| 30. |
- Pathan, M., et al.
(författare)
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A novel community driven software for functional enrichment analysis of extracellular vesicles data
- 2017
-
Ingår i: Journal of Extracellular Vesicles. - : Wiley. - 2001-3078. ; 6:1
-
Tidskriftsartikel (refereegranskat)abstract
- Bioinformatics tools are imperative for the in depth analysis of heterogeneous high-throughput data. Most of the software tools are developed by specific laboratories or groups or companies wherein they are designed to perform the required analysis for the group. However, such software tools may fail to capture "what the community needs in a tool". Here, we describe a novel community-driven approach to build a comprehensive functional enrichment analysis tool. Using the existing FunRich tool as a template, we invited researchers to request additional features and/or changes. Remarkably, with the enthusiastic participation of the community, we were able to implement 90% of the requested features. FunRich enables plugin for extracellular vesicles wherein users can download and analyse data from Vesiclepedia database. By involving researchers early through community needs software development, we believe that comprehensive analysis tools can be developed in various scientific disciplines.
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| 31. |
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| 32. |
- Ronquist, Göran, et al.
(författare)
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Antibody-Loading of Biological Nanocarrier Vesicles Derived from Red-Blood-Cell Membranes
- 2024
-
Ingår i: ACS Omega. - 2470-1343. ; 9, s. 22711–22718-
-
Tidskriftsartikel (refereegranskat)abstract
- Antibodies, disruptive potent therapeutic agents against pharmacological targets, face a barrier in crossing immune systems and cellular membranes. To overcome these, various strategies have been explored including shuttling via liposomes or biocamouflaged nanoparticles. Here, we demonstrate the feasibility of loading antibodies into exosome-mimetic nanovesicles derived from human red-blood-cell membranes, which can act as nanocarriers for intracellular delivery. Goat-antichicken antibodies are loaded into erythrocyte-derived nanovesicles, and their loading yields are characterized and compared with smaller dUTP-cargo molecules. Applying dual-color coincident fluorescence burst analyses, the loading yield of nanocarriers is rigorously profiled at the single-vesicle level, overcoming challenges due to size-heterogeneity and demonstrating a maximum antibody-loading yield of 38-41% at the optimal vesicle radius of 52 nm. The achieved average loading yields, amounting to 14% across the entire nanovesicle population, with more than two antibodies per loaded vesicle, are fully comparable to those obtained for the much smaller dUTP molecules loaded in the nanovesicles after additional exosome-spin-column purification. The results suggest a promising new avenue for therapeutic delivery of antibodies, potentially encompassing also intracellular targets and suitable for large-scale pharmacological applications, which relies on the exosome-mimetic properties, biocompatibility, and low-immunogenicity of bioengineered nanocarriers synthesized from human erythrocyte membranes.
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| 33. |
- Ronquist, Gunnar, et al.
(författare)
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Captopril may reduce biochemical (prostate-specific antigen) failure following radical prostatectomy for clinically localized prostate cancer
- 2009
-
Ingår i: Scandinavian Journal of Urology and Nephrology. - : Informa UK Limited. - 0036-5599 .- 1651-2065. ; 43:1, s. 32-36
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: A prior report suggested that individuals medicated with captopril showed a decreased incidence of prostate cancer. This study therefore investigated whether captopril given postoperatively had any preventive effect on biochemical recurrence for patients treated with radical prostatectomy. MATERIAL AND METHODS: Data were prospectively reviewed for 62 men subjected to radical retropubic prostatectomy due to biopsy-confirmed, clinically localized prostate cancer and comparisons were made between two groups, those receiving captopril postoperatively (12.5 mg twice daily; captopril group, n=32) and those not receiving any captopril (control group, n=30). One surgeon carried out the surgery. RESULTS: The two groups were comparable as regards age at surgery, prostate volume, preoperative prostate-specific antigen values, pathological stage, Gleason score, organ-confined disease, occurrence of positive surgical margins and extraprostatic extension. The incidence of biochemical failure was three out of 32 patients in the captopril group and 10 out of 30 in the control group (p=0.034) during a mean observational time of 29 months. CONCLUSIONS: A lower rate of biochemical recurrence was observed in men subjected to radical prostatectomy treated with captopril postoperatively than in those not receiving captopril. These results were based on only 32 observations; a larger study may show no evidence of an association.
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| 34. |
- Ronquist, Göran
(författare)
-
Extracellular vesicles and energy metabolism
- 2019
-
Ingår i: Clinica Chimica Acta. - : Elsevier BV. - 0009-8981 .- 1873-3492. ; 488, s. 116-121
-
Forskningsöversikt (refereegranskat)abstract
- Glycolytic enzymes are among the most frequently identified proteins in proteomics of exosomes/extracellular vesicles. This review brings up the possibility that exosomes/extracellular vesicles during their life-time in bodily fluids power important energy-consuming functions by glycolytic conversion of glucose or fructose into ATP. It was seen that prostasomes (exosomes of the prostate) could produce ATP by glycolysis and that the produced ATP quickly was consumed by adjacent prostasomal ATPases. The glycolytic ATP production appeared to be coupled to self-sustaining energy requirements. It will also be discussed how a failure in this machinery (lowered activity of ATPases) with a resultant polluting leakage of extracellular ATP could affect cancer development.
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| 35. |
- Ronquist, Göran, 1971-
(författare)
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Some Characteristics of Human Prostasomes and Their Relationship to Prostate Cancer
- 2009
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Background: The secretory epithelial cells of the prostate gland use sophisticated vehicles named prostasomes to relay important information to sperm cells in semen. This prostasome-forming and secretory ability of the epithelial cells is also preserved in poorly differentiated prostate cancer cells. Aim: The aim of this thesis was to examine different characteristics of prostasomes, especially those derived from malignant prostate cells, linked to their potential role in diagnosis and prognostication of prostate cancer. Results: Serum samples of prostate cancer patients contained autoantibodies against seminal prostasomes in a higher concentration than did control sera. These autoantibodies were most frequently directed against 25 prostasome-associated proteins, but no one was prostate specific. Clusterin was one of the most frequently occurring prostasomal proteins. Elevated titers were however seen in both patients´ and control sera. Clusterin turned out to be a major antigen of seminal prostasomes. No prostate specific or prostate cancer specific protein was discovered upon proteomic analysis of prostasomes deriving from malignant cells of vertebral metastases of prostate cancer patients. Human chromosomal DNA was identified in both seminal prostasomes and PC-3 cell prostasomes and strong evidence existed that the DNA was localized inside the prostasomes. Four out of 13 DNA clones of seminal prostasomes featured gene sequences (31%). The corresponding figures for PC-3 cell prostasomes were 4 out of 16 clones (25%). Conclusions: Prostasomes are immunogenic and give rise to serum autoantibodies. The most frequently occurring autoantibodies were directed against 25 prostasomal proteins but none of these was exclusively prostate specific. Thirty different proteins were identified in prostate cancer metastasis-derived prostasomes but none of these proteins was prostate cancer specific. Human chromosomal DNA was identified in prostasomes of both normal and malignant cell origin.
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| 36. |
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| 37. |
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| 38. |
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| 39. |
- Sahlén, Göran, 1957-
(författare)
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Formation,Storage and Secretion of Prostasomes in Benign and Malignant Cells and Their Immunogenicity in Prostate Cancer Patients
- 2007
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Prostasomes are submicron-sized, membrane-bound organelles produced by the epithelial cells of the prostate and normally found in the secretion in the gland ducts. Their physiological role is in the promotion of sperm-function in human reproduction. This thesis contains four papers dealing with the production of prostasomes and some possible applications in clinical urology of the prostasome. Paper I and II provided an ultrastructural description of the synthesis, storage and secretion of prostasomes in benign as well as in malignant tissue. Most notable were the extracellular appearances of prostasomes in metastatic lesions whereby the prostasomes become exposed to the immune system of the patient. This supported findings in earlier studies in which patients with advanced prostate cancer had elevated levels of anti-prostasome antibodies. The results of paper III reinforced the view of the prostate-unique origin of the prostasome. In particular, there were no indications in SDS-PAGE patterns or flow-cytometric studies of material from seminal vesicle secretion that it contained components that could be associated with a production of prostasomes. Some possible clinical functions of the prostasomes were investigated in paper IV. Exposure of prostasomes to the immune system through mechanical and thermal trauma to the prostate did not induce an evident formation of anti-prostasome autoantibodies. Furthermore, the serum levels of anti-prostasome antibodies registered by assays with preparations of prostasomes from seminal plasma as antigen did not correlate with existing prostate cancer. Seminal prostasomes seemed not to function as substitute markers for prostate cancer in the test kit used. A possible explanation could be underestimated differences in antigen properties between seminal or prostate gland-derived prostasomes and prostasomes from tumor tissue.
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| 40. |
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| 41. |
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| 42. |
- Sahlén, Göran, et al.
(författare)
-
Secretions from seminal vesicles lack characteristic markers for prostasomes
- 2010
-
Ingår i: Upsala Journal of Medical Sciences. - : Uppsala Medical Society. - 0300-9734 .- 2000-1967. ; 115:2, s. 107-112
-
Tidskriftsartikel (refereegranskat)abstract
- Background. Prostasomes are suggested to be produced in the prostate gland. Although biochemical studies support this, some immunohistochemical findings indicate that also the seminal vesicles could be a source of prostasomes. Therefore, we have compared the secretion of the vesicles with that of the prostate using biochemical and ultrastructural techniques. Methods. Ultracentrifuged pellets of substance from seminal vesicle secretions were analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and flow cytometry. The secretory cells of the seminal vesicles were examined with transmission electron microscopy. These findings were then compared with published results from similar studies of the prostate secretory cells. Results. In SDS-PAGE, the seminal vesicle pellets lacked the three prostasome-characteristic CD-markers, namely CD10, CD13, and CD26, but expressed two proteins of about 55 kDa and 70 kDa, corresponding to clusterin and heat shock protein (HSP70). Flow cytometry showed the presence of secretion particles in the seminal pellet, although of a smaller size than that of the prostasomes. Electron microscopy of the luminal part of the cells in the seminal vesicles demonstrated many secretion granules, each enclosed in a vesicle with a size of about 1 mum. Conclusions. Pelleted seminal vesicle secretion is different to prostate secretion in several ways. No prostasome characteristics were detected in the pelleted seminal vesicle secretion.
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| 43. |
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| 44. |
- Sanaee, Maryam, et al.
(författare)
-
Antibody-loading of biological nanocarrier vesicles derived from red-blood-cell membranes
-
Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Antibodies, disruptive potent therapeutic agents against pharmacological targets, face a barrier crossing immune-system and cellular-membranes. To overcome these, various strategies have been explored including shuttling via liposomes or bio-camouflaged nanoparticles.Here, we demonstrate the feasibility to load antibodies into exosome-mimetic nanovesicles derived from human red-blood-cell-membranes. The goat-anti-chicken antibodies are loaded into erythrocyte-membrane derived nanovesicles and their loading yields are characterized and compared with smaller dUTP-cargo. Applying dual-color coincident fluorescence burst methodology, the loading yield of nanocarriers is profiled at single-vesicle level overcoming their size-heterogeneity and achieving a maximum of 38-41% antibody-loading yield at peak radius of 52 nm. The average of 14 % yield and more than two antibodies per vesicle is estimated, comparable to those of dUTP-loaded nanovesicles after additional purification through exosomespin-column. These results suggest a promising route for enhancing biodistribution andintracellular accessibility for therapeutic antibodies using novel, biocompatible, and lowimmunogenicity nanocarriers, suitable for large-scale pharmacological applications.
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| 45. |
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| 46. |
- Sanaee, Maryam, et al.
(författare)
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Dual-Color Confocal Fluorescence Characterizations of Antibody Loading in Bioengineered Nanovesicles
- 2023
-
Ingår i: 2023 Conference on Lasers and Electro-Optics Europe and European Quantum Electronics Conference, CLEO/Europe-EQEC 2023. - : Institute of Electrical and Electronics Engineers (IEEE).
-
Konferensbidrag (refereegranskat)abstract
- Confocal fluorescence microscopy is powerful for microscopic and nanoscopic analyses with a broad range of applications in biology and medicine, affording high specificity and sensitivites down to single-molecule level [1]. Here we apply a novel dual-color confocal fluorescence microscopy methodology based on coincident burst analysis [2] to quantitative study of the loading yields of bioengineered nanovesicles derived from human red blood cell (RBC) membranes loaded with fluorescently-tagged Antibody (Ab) or dUTP cargo molecules. We prove the successful loading of the RBC nanovesicles with both types of cargo molecules, assess their size statistics and provide quantifications of the loading at single-vesicle level. Fig. 1a shows the tagging scheme adopted for the experiment where Cellvue claret and Alexa488 dyes are respectively used to fluorescently label the outer membrane of the nanovesicles and the cargo molecules. The experimental setup is shown in Fig. 1b, consisting of an inverted microscope with dual excitations at λ1= 640 nm and λ2 = 485 nm. The fluorescence signals centered at λred= 720 nm and λgreen= 535 nm are single-photon-detected and post-selected in separate time gating windows with a delay of 25ns (Fig. 1c). After looking for coincident red-green bursts in the measured time traces (Fig. 1d), size histograms of the loaded vesicles are retrieved (Fig. 1e-f), obtaining average loading yields of 5% and 20% for Ab and dUTP cargos, respectively. Average values of 1.5 and 1.4 loaded Ab (dUTP) cargo molecules per nanovesicle are retrieved from the histograms of Fig. 1g (Fig. 1h). The average size of Ab-loaded nanovesicles is found to be slightly (≈10 nm in radius) larger than the one of dUTP-loaded ones.
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| 47. |
- Sanaee, Maryam, et al.
(författare)
-
Size stability and self-agglomeration of erythrocyte-derived membrane nanovesicles versus physiological extracellular vesicles
-
Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Extracellular vesicles (EVs) and plasma membrane-derived exosome-mimetic nanovesicles demonstrate significant potential for drug delivery. Latter synthetic provides higher throughput over physiological EVs. However they face size-stability and self-agglomeration challenges in physiological solutions to be properly characterized and addressed. Here we demonstrate a fast and high-throughput nanovesicle screening methodology relying on dynamic light scattering (DLS) complemented by atomic force microscopy (AFM)measurements, suitable for the evaluation of hydrodynamic size instabilities and aggregation effects in nanovesicle solutions under varying experimental conditions and apply it to the analysis of bio-engineered nanovesicles derived from erythrocytes as well as physiological extracellular vesicles isolated from animal seminal plasma. The synthetic vesicles exhibit a significantly higher degree of agglomeration, with only 8 % of them falling within the typical extracellular vesicle size range (30-200 nm) in their original preparation conditions. Concurrent zeta potential measurements performed on both physiological and syntheticnanovesicles yielded values in the range of -17 to -22 mV, with no apparent correlation to their agglomeration tendencies. However, mild sonication and dilution were found to be effective means to restore the portion of EVs-like nanovesicles in synthetic preparations to values of 54% and 63%, respectively, The results illustrate the capability of this DLS-AFM-based analytical method for real-time, high-throughput and quantitative assessments of agglomeration effects and size instabilities in bioengineered nanovesicle solutions, providing a powerful and easy-to-use tool to gain insights to overcome such deleterious effects and leverage the full potential of this promising biocompatible drug-delivery carriers for a broad range of pharmaceutical applications.
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| 48. |
- Stark, Katarina, et al.
(författare)
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Expression of CYP4F12 in gastrointestinal and urogenital epithelia
- 2004
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Ingår i: Basic & Clinical Pharmacology & Toxicology. - : Wiley. - 1742-7835 .- 1742-7843. ; 94:4, s. 177-83
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Tidskriftsartikel (övrigt vetenskapligt/konstnärligt)abstract
- Cytochrome P450 4F12 (CYP4F12) was originally cloned from human liver and small intestine. CYP4F12 can oxidize arachidonic acid, two stable prostaglandin H2 analogues, and an antihistamine, ebastine, but the tissue distribution and catalytic properties of CYP4F12 have not been fully investigated. An antipeptide polyclonal antibody was raised against the C-terminal of CYP4F12 (PLNVGLQ), evaluated by Western blot analysis and used for immunohistological analysis of 50 human tissues. Western blot analysis of recombinant CYP4F12, expressed in yeast, and microsomal proteins from adult and foetal liver, kidney, placenta at term, seminal vesicles, the prostate gland and purified prostasomes showed that the polyclonal antibody detected a protein of the expected size, approximately 60 kDa. CYP4F12 mRNA could be detected in seminal vesicles and prostate gland by reverse transcription-PCR. Prominent CYP4F12 immunoreactivity occurred, inter alia, in the epithelial cells of the gastrointestinal tract (stomach, small intestine, and colon), collecting tubules, transitional epithelium, ovarian follicles, the endothelium of microvessels of placental villi (first trimester), and epidermis. We screened recombinant CYP4F12 for catalytic activity. Arachidonic acid (20:4n-6) was hydroxylated at C18 and laurate at C11, but significant amounts of metabolites of 18:2n-6, 20:3n-9, 20:5n-3, 22:5n-6, and some prostaglandins could not be detected. We conclude that CYP4F12 is widely distributed in gastrointestinal and urogenital epithelia and exhibits a narrow substrate specificity.
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| 49. |
- Vårdal, Hege, 1972-
(författare)
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From Parasitoids to Gall Inducers and Inquilines : Morphological Evolution in Cynipoid Wasps
- 2004
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- One of the large lineages of parasitic wasps, the Cynipoidea, exhibits three distinctly different life modes. Slightly more than half of the about 3000 species are parasitoids in insect larvae, whereas the remaining species are associated with plants, either as gall inducers or as inquilines (guests feeding on plant tissue in galls). The main focus of this thesis has been to identify morphological changes associated with the shifts between life modes. Particular attention was paid to structures believed to be important in gall initiation. Comparative anatomical studies of the egg, larva and venom apparatus were performed, including representatives of parasitoids, gall inducers and inquilines. Examination of gross morphology and ultrastructure revealed that the eggs of the gall inducers are larger and surrounded by a thicker shell than the parasitoid eggs. These differences may be related to the fact that the gall inducer egg contains sufficient egg yolk for the embryo during the entire egg period, whereas the parasitoid egg often absorbs nutrients through the eggshell. Furthermore, the gall inducer egg is probably more exposed to desiccation and therefore a thicker and more resistant eggshell is crucial. Comparing the terminal-instar larvae of about 30 species of parasitoids, gall inducers and inquilines, extensive morphological variation was found, particularly in the head and mouthpart features. The variation was summarized in 33 morphological and one life-history character and parsimony analyses were performed. The resulting phylogenetic estimates were largely in accordance with previous analyses of adult morphology and molecular data. The larval data point to a single origin of the inquilines, in agreement with adult morphology but in conflict with molecular data. The venom apparatus was found to be quite uniform in structure among a sample of 25 species of cynipoid species. It consists of a very short venom duct, a reservoir and a single unbranched venom gland. With few exceptions, the venom apparatus is conspicuously larger relative to the female metasoma in the gall inhabiting species than in the parasitoids. We found little evidence of anatomical structures that could facilitate chemical communication between the gall-inducer embryo and the surrounding plant tissue through the thick eggshell. On the other hand, the enormous venom glands and reservoirs, which are apparently not used for defence, suggest that the adult female plays a significant role in gall induction by injecting secretions into the host plant when laying eggs.
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| 50. |
- Vårdal, Hege, et al.
(författare)
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Morphology and evolution of the cynipoid egg (Hymenoptera)
- 2003
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Ingår i: Zoological Journal of the Linnean Society. - London : Blackwell Publishing. - 0024-4082 .- 1096-3642. ; 139:2, s. 247-260
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Tidskriftsartikel (refereegranskat)abstract
- We describe gross egg morphology and provide the first data on eggshell ultrastructure in cynipoids (Hymenoptera) based on species representing three distinctly different life histories: internal parasitoids of endopterygote larvae, gall inducers and phytophagous inquilines (guests in galls). We then use existing phylogenetic hypotheses to identify putative changes in egg structure associated with evolutionary life-history transitions. We find four major structural changes associated with the shift from parasitoids laying their eggs inside a host larva to gall inducers laying their eggs in or on plants: (1) from a narrow and gradually tapering gross form to a distinct division into a stout body and a long and thin stalk; (2) from a thin to a thick eggshell; (3) from a flexible to a rigid endochorion; and (4) from crystal bundles with shifting orientation in the exochorion to layers of parallel crystal rods. By contrast, we find no major changes in egg structure associated with the transition from gall inducers to inquilines. Comparison between pre- and post-oviposition eggs of one gall inducer and one inquiline suggests that mechanical stress during the passage through the egg canal gives rise to numerous tiny stress fractures in the boundary separating the exo- and endochorion. In one of the gall inducers, Diplolepis rosae, that end of the egg, which is inserted into the plant, has a specialized and apparently porous shell that may permit chemical exchange between the embryo and the plant. Other structures that could facilitate chemical communication with the host plant through the eggshell were, however, not observed in the eggs of gall inhabitants.
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