143 results on '"Vuento M"'
Search Results
2. Detection of canine parvovirus antigens with antibodies to synthetic peptides
- Author
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Vihinen-Ranta, M., Lindfors, E., Heiska, L., Veijalainen, P., and Vuento, M.
- Published
- 1996
- Full Text
- View/download PDF
3. CELL FUSION IN ELECTRIC FIELD IN THE PRODUCTION OF MONOCLONAL ANTIBODIES AGAINST SOLUBLE AND CELLULAR POLYPEPTIDE ANTIGENS
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VUENTO, M., primary, NIKKILÄ, L., additional, and RAUVALA, H., additional
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- 1987
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4. Reconstruction of the Vulva with Sensate Gluteal Fold Flaps
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Kuokkanen, H., primary, Mikkola, A., additional, Nyberg, R. H., additional, Vuento, M. H., additional, Kaartinen, I., additional, and Kuoppala, T., additional
- Published
- 2013
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5. P15.05: Vascularity measurements in predicting deep invasion in endometrial carcinoma: uterine artery color Doppler and uterine 3D‐power Doppler angiography
- Author
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Saarelainen, S. K., primary, Vuento, M. H., additional, Kirkinen, P., additional, and Mäenpää, J. U., additional
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- 2012
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6. Monitoring human parvovirus B19 virus-like particles and antibody complexes in solution by fluorescence correlation spectroscopy
- Author
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Toivola, J., primary, Michel, P. O., additional, Gilbert, L., additional, Lahtinen, T., additional, Marjomäki, V., additional, Hedman, K., additional, Vuento, M., additional, and Oker-Blom, C., additional
- Published
- 2004
- Full Text
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7. Assembly of fluorescent chimeric virus-like particles of canine parvovirus in insect cells
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Gilbert, L, primary, Toivola, J, additional, Lehtomäki, E, additional, Donaldson, L, additional, Käpylä, P, additional, Vuento, M, additional, and Oker-Blom, C, additional
- Published
- 2004
- Full Text
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8. Properties of Baculovirus Particles Displaying GFP Analyzed by Fluorescence Correlation Spectroscopy
- Author
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Toivola, J., primary, Ojala, K., additional, Michel, P.O., additional, Vuento, M., additional, and Oker-Blom, C., additional
- Published
- 2002
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9. Endometrial fluid accumulation in asymptomatic postmenopausal women
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Vuento, M. H., primary, Pirhonen, J. P., additional, Mäkinen, J. I., additional, Tyrkkö, J. E., additional, Laippala, P. J., additional, Gröroos, M., additional, and Salmi, T. A., additional
- Published
- 1996
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10. Staining of Immunoblots Immunochromatography
- Author
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Herranen, M., primary and Vuento, M., additional
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- 1994
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11. Rapid detection of methicillin-resistant Staphylococcus aureus strains not identified by slide agglutination tests
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Kuusela, P, primary, Hildén, P, additional, Savolainen, K, additional, Vuento, M, additional, Lyytikäinen, O, additional, and Vuopio-Varkila, J, additional
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- 1994
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12. Evaluation of Three Slide Agglutination Tests for Rapid Identification of Staphylococcus aureus
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Niskanen, A., primary, Korkeala, H., additional, Manninen, M., additional, Vuento, M., additional, and Kuusela, P., additional
- Published
- 1991
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13. A longitudinal study of screening for endometrial cancer by endometrial biopsy in diabetic females.
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VUENTO, M. H., MAATELA, J. I., TYRKKÖ, J. E., LAIPPALA, P. J., GRÖNROOS, M., and SALMI, T. A.
- Abstract
Diabetics are at high risk of developing endometrial cancer; the relative risk of endometrial cancer in diabetics is fourfold in comparison to non-diabetic controls. The purpose of this longitudinal study was to evaluate the effectiveness of screening asymptomatic diabetic females in terms of the premalignant and malignant endometrial findings, and to try to determine the optimal screening interval. In 1980-1981, a group of 462 diabetic females was identified and registered. One half of them (237) was invited to be screened. Endometrial samples were taken by using Vabra aspiration. The results of this first randomized screening in 1980-1981 have been published elsewhere. At that time 124 females participated. The remaining 225 females acted as an unscreened control group. Eight years later (1988-1989), both groups were invited to be screened. The Pistolet aspiration method was used. At this stage, group 1 (screened in 1980-1981) consisted of 78 females, and group 2 (not screened in 1980-1981) consisted of 148 females. In 85% (193/226) of the females, the uterine cavity was reached with the Pistolet instrument; 96% of the females found the pain acceptable. In the group screened twice (group 1), no pathologic lesions of the endometrium were found in the second screening. In the group screened for the first time (group 2), one female had endometrial adenocarcinoma (0.8%), one had complex hyperplasia without atypia (0.8%) and four had endometrial polyps (3.3%). In 165 cases of 193, both a cytologic and a histologic specimen were available. In 130 cases (79%) the cytology was of class I, including the one endometrial adenocarcinoma. In three cases (2%) it was of class II and in one case (1%) of class III. Endometrial biopsy by Pistolet aspiration was a method highly acceptable by the patients for examining the endometrium. However, cytologic examination was not able to show the existing endometrial adenocarcinoma. One endometrial sampling of asymptomatic diabetic females during early menopause could detect the bulk of the occult, slowly progressing lesions of the endometrium. Such screeening might be most efficient in terms of cost-benefit ratio. [ABSTRACT FROM AUTHOR]
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- 1995
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14. A monkey antigen crossreacting with carcinoembryonic antigen, CEA*.
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Engvall, E, Vuento, M, and Ruoslahti, E
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- 1976
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15. Fibronectin fragmentation induced by dental plaque and Bacteroides gingivalis.
- Author
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LARJAVA, H., UITTO, V-J., HAAPASALO, M., HEINO, J., and VUENTO, M.
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- 1987
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16. Evaluation of ovarian findings in asymptomatic postmenopausal women with color Doppler ultrasound.
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Vuento, Maarit H., Pirhonen, Jouko P., Mäkinen, Juha I., Laippala, Pekka J., Grönroos, Matti, Salmi, Tuula A., Vuento, M H, Pirhonen, J P, Mäkinen, J I, Laippala, P J, Grönroos, M, and Salmi, T A
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- 1995
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17. Mass screening for endometrial cancer directed in risk groups of patients with diabetes and patients with hypertension.
- Author
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Grönroos, Matti, Salmi, Tuula A., Vuento, Maarit H., Jalava, E. Anneli, Tyrkkö, Juhani E., Maatela, Jouni I., Aromaa, Arpo R., Siegberg, Rita, Savolainen, Eeva R., Kauraniemi, Tapani V., Grönroos, M, Salmi, T A, Vuento, M H, Jalava, E A, Tyrkkö, J E, Maatela, J I, Aromaa, A R, Siegberg, R, Savolainen, E R, and Kauraniemi, T V
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- 1993
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18. Fibronectin fragmentation induced by dental plaque and Bacteroides gingivalis
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Vuento M, Veli-Jukka Uitto, Jyrki Heino, Markus Haapasalo, and Hannu Larjava
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Dental Plaque ,Dental plaque ,Microbiology ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Bacteroides ,Humans ,Trypsin ,Fragmentation (cell biology) ,General Dentistry ,030304 developmental biology ,0303 health sciences ,biology ,Chemistry ,Elastase ,Biological activity ,030206 dentistry ,medicine.disease ,biology.organism_classification ,Pepsin A ,In vitro ,Fibronectins ,Fibronectin ,Gelatinases ,biology.protein ,medicine.drug - Abstract
– Degradation of fibronectin (FN) by subgingival and supragingival plaque and Bacteroides gingivalis (Bg) was studied in vitro. The degradation of FN by both types of plaque was relatively rapid, continuous but incomplete. Some differences were found between supra-and subgingival samples. Supragingival plaque extracts produced several FN fragments of 110–180 kd during short incubations, of 15–60 min. The predominant fragment after overnight incubation was a 110 kd polypeptide. With subgingival plaque extract a more extensive degradation of FN was noted. The main degradation product was a 120 kd fragment after overnight incubation. Several peptide fragments were released from fibronectin by Bg extracts. Their molecular size was different from those produced by trypsin, elastase or dental plaque. When cell extracts of Bg were fractionated by high performance liquid chromatography, three separate peaks of fibronectin degrading activity were obtained. Two of those peaks also contained trypsin-like enzyme activity. The degradation of fibronectin and the subsequent formation of biologically active peptides may have many effects in periodontal pockets. These may include modifying effects on plaque growth and wound healing.
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- 1987
19. Identification of fibronectin fragments that bind to carboxy-group-modified proteins
- Author
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Vuento, M, Sekiguchi, K, and Korkolainen, M
- Abstract
Limited proteolysis of human plasma fibronectin with chymotrypsin, trypsin or thermolysin has been used to localize binding sites responsible for binding [Vuento, Korkolainen & Stenman (1982) Biochem. J. 205, 303-311] of fibronectin to carboxy-group-modified proteins. These bindings sites are different from those mediating binding of fibronectin to gelatin or heparin. They are located close to the C-terminus of the polypeptide chains of fibronectin, and apparently overlap with the C-terminal fibrin binding site.
- Published
- 1983
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20. Attachment of staphylococci and streptococci on fibronectin, fibronectin fragments, and fibrinogen bound to a solid phase
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Kuusela, P, Vartio, T, Vuento, M, and Myhre, E B
- Abstract
The attachment of Staphylococcus aureus (Cowan I) and two strains of group A and G streptococci on glass cover slips coated with fibronectin, fibronectin fragments, or fibrinogen was studied. The attachment was quantitated by counting the attached bacteria on glass surfaces coated with a similar molarity of the proteins. Fibronectin was a more effective attachment factor than fibrinogen for staphylococci, while group G streptococci attached better on fibrinogen- than on fibronectin-coated cover slips. In this system, group A streptococci bound almost exclusively to substrate-bound fibrinogen. Attachment experiments involving the use of staphylococci pretreated with soluble fibronectin or fibrinogen revealed that bacterium-bound fibronectin and fibrinogen were able to enhance the adherence on cover slips coated with fibronectin. The 30-kilodalton NH2-terminal and the 120- to 140-kilodalton COOH-terminal fragments of fibronectin, both of which contain bacterial binding sites, mediated the staphylococcal attachment, suggesting that both parts of the molecule are involved in the attachment mediated by fibronectin.
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- 1985
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21. Binding sites for streptococci and staphylococci in fibronectin
- Author
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Kuusela, P, Vartio, T, Vuento, M, and Myhre, E B
- Abstract
Purified cathepsin G fragments of fibronectin were used to locate the binding sites for streptococci and staphylococci in the fibronectin molecule. The iodinated, NH2-terminal, 30-kilodalton (kd) fragment bound to group A and G streptococci and to Staphylococcus aureus. The 125I-labeled, COOH-terminal, 120- to 140-kd fragment bound weakly to group A streptococcus strain and to S. aureus when tested in a buffer of low ionic strength. The 30- and 120- to 140-kd fragments inhibited the binding of iodinated fragments to bacteria. The two fragments were, on a molar basis, equally effective, and they were more potent inhibitors than intact fibronectin. The gelatin-binding 40-kd fragment neither bound to any of the bacterial strains nor inhibited the binding of 125I-labeled 30-kd or 125I-labeled 120- to 140-kd fragments to bacteria. The results indicate that fibronectin has at least two separate binding sites for streptococci and staphylococci, one in the NH2-terminal region and another in the COOH-terminal region of the molecule, both capable of specific interaction with a complementary structure exposed on streptococcal and staphylococcal cell surfaces.
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- 1984
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22. Isolation of a novel cell-attachment and spreading-promoting protein from human serum
- Author
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Vuento, M, Korkolainen, M, Kuusela, P, and Hölttä, E
- Abstract
A protein with potent cell-attachment and spreading-promoting activity was isolated from fibronectin-free human serum. The purification steps included affinity chromatography on heparin-agarose and preparative isoelectric focusing. The purified protein was homogeneous as judged from dodecyl sulphate/polyacrylamide-gel electrophoresis. It had an isoelectric point of 5.0 and an Mr of 52 000. The protein promoted the spreading of Chinese-hamster ovary cells to plastic in a manner similar to that observed with fibronectin.
- Published
- 1985
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23. A monkey antigen crossreacting with carcinoembryonic antigen, CEA*
- Author
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Engvall, E, Vuento, M, and Ruoslahti, E
- Abstract
Normal monkey tissues were found to contain an antigen which crossreacts immunologically with the carcinoembryonic antigen (CEA) of the human digestive tract. The monkey antigen reacted with complete or partial identity to the normal crossreacting antigen (NCA) in humans when tested in immunodiffusion against anti-CEA or anti-NCA. Extracts of monkey tissues inhibited in radioimmunoassays measuring human NCA. It is possible that monkey foetuses and colonic tumours contain CEA.
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- 1976
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24. Essential charged amino acids in the binding of fibronectin to gelatin
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Vuento, M, Salonen, E, Osterlund, K, and Stenman, U H
- Abstract
The binding of fibronectin to gelatin-agarose was strictly dependent on pH, having a pH optimum of 7-9. The binding was strongly inhibited by increasing ionic strength. A chemical modification of lysyl and arginyl groups of fibronectin abolished the binding activity. The anionic detergents sodium dodecyl sulphate and sodium deoxycholate in concentrations of 10-100mM had the same effect. The binding was not affected by the non-ionic detergents Triton X-100, Tween 20 or Lubrol WX. The results demonstrate an important role of ionic interactions in the binding of fibronectin to gelatin. Absence of inhibition by non-ionic detergents suggests that hydrophobic interactions contribute relatively little to the binding of fibronectin to gelatin.
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- 1982
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25. Immunochemical characterization of human plasma fibronectin
- Author
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Vuento, M, Salonen, E, Salminen, K, Pasanen, M, and Stenman, U K
- Abstract
Human plasma fibronectin has been purified by a non-denaturing affinity chromatography procedure [Vuento & Vaheri, (1979) Biochem.J. 183, 331–337], and antisera have been raised by immunizing rabbits with the native protein. The antisera reacted strongly with native fibronectin, but only weakly with reduced and alkylated fibronectin or with heat-denaturated fibronectin. Denaturation also affected the haemagglutinating and gelatin-binding activities of fibronectin and increased its susceptibility to proteolytic degradation. The antisera reacted with fragments of fibronectin obtained by proteolysis with plasmin. Large fragments (mol.wt. 180000–200000), lacking the region harbouring the interchain disulphide bridges but containing the sites responsible for gelatin-binding and haemagglutinating activity, showed as intense a reaction with the antisera as intact fibronectin. Smaller peptides showed a weaker reaction. All fragments tested showed sensitivity to denaturation in their reaction with the antisera. The results were interpreted as showing that: (1) native fibronectin has an ordered conformation that is easily perturbed by denaturation; (2) most of the antigenic determinants of the protein are dependent on conformation; (3) the region of the fibronectin molecule containing the interchain disulphide bridges has only few antigenic determinants; and (4) covalent interaction of the two subunits does not contribute to the antigenic structure recognized by rabbit antisera. The observed correlation between the antigenic activity and a structural and functional intactness of fibronectin suggests that the antibodies to native fibronectin could be used as a conformational probe in studies on this protein.
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- 1980
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26. Purification of fibronectin from human plasma by affinity chromatography under non-denaturing conditions
- Author
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Vuento, M and Vaheri, A
- Abstract
Fibronectin was purified from human plasma by affinity chromatography under nondenaturing conditions. The method was based on the previously known binding of fibronectin to gelatin. The novel features of our method are the use of arginine in the elution of fibronectin from immobilized gelatin [Vuento & Vaheri (1978) Biochem. J. 175, 333-336] and the use of arginine-agarose as second affinity step. The purified protein was homogeneous as judged by polyacrylamide-gel electrophoresis, analytical ultracentrifugation and two-dimensional immunoelectrophoresis. The yield was 60%. We propose that the method would be useful in preparation of fibronectin for studies on its biological activities, where it is important that the protein is obtained in a native state.
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- 1979
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27. Dissociation of fibronectin from gelatin-agarose by amino compounds
- Author
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Vuento, M and Vaheri, A
- Abstract
Soluble fibronectin of human plasma was specifically dissociated at neutral pH from gelatin-agarose by several cationic amino compounds, notably the polyamines spermine, spermidine and putrescine, the basic amino acid arginine, and amino sugars. The neutral and acidic amino acids and the N-acetylated derivatives of amino sugars tested were ineffective. Gel-filtration experiments demonstrated that [14C]spermidine bound to fibronectin but not to gelatin.
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- 1978
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28. Affinity immunosensor for milk progesterone: identification of critical parameters
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Laitinen, M. P. A. and Vuento, M.
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- 1996
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29. Purification of the Carcinoembryonic Antigen (CEA) with Immunoadsorbents
- Author
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VUENTO, M., primary, ENGVALL, E., additional, and RUOSLAHTI, E., additional
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- 1976
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30. Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like particles
- Author
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Vuento Matti, Korhonen Eila, Mäkelä Anna R, White Daniel, Cunningham Claire, Saloniemi Taija, Välilehto Outi, Toivola Jouni, Gilbert Leona, and Oker-Blom Christian
- Subjects
Biotechnology ,TP248.13-248.65 ,Medical technology ,R855-855.5 - Abstract
Abstract Fluorescence correlation spectroscopy (FCS) monitors random movements of fluorescent molecules in solution, giving information about the number and the size of for example nano-particles. The canine parvovirus VP2 structural protein as well as N-terminal deletion mutants of VP2 (-14, -23, and -40 amino acids) were fused to the C-terminus of the enhanced green fluorescent protein (EGFP). The proteins were produced in insect cells, purified, and analyzed by western blotting, confocal and electron microscopy as well as FCS. The non-truncated form, EGFP-VP2, diffused with a hydrodynamic radius of 17 nm, whereas the fluorescent mutants truncated by 14, 23 and 40 amino acids showed hydrodynamic radii of 7, 20 and 14 nm, respectively. These results show that the non-truncated EGFP-VP2 fusion protein and the EGFP-VP2 constructs truncated by 23 and by as much as 40 amino acids were able to form virus-like particles (VLPs). The fluorescent VLP, harbouring VP2 truncated by 23 amino acids, showed a somewhat larger hydrodynamic radius compared to the non-truncated EGFP-VP2. In contrast, the construct containing EGFP-VP2 truncated by 14 amino acids was not able to assemble into VLP-resembling structures. Formation of capsid structures was confirmed by confocal and electron microscopy. The number of fluorescent fusion protein molecules present within the different VLPs was determined by FCS. In conclusion, FCS provides a novel strategy to analyze virus assembly and gives valuable structural information for strategic development of parvovirus-like particles.
- Published
- 2006
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31. Role of mitochondria in parvovirus pathology.
- Author
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Nykky J, Vuento M, and Gilbert L
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- Animals, Calcium metabolism, Cats, Cell Line, Dogs, Enzyme Activation, Extracellular Signal-Regulated MAP Kinases metabolism, MAP Kinase Signaling System, Membrane Potential, Mitochondrial, Mitochondria pathology, Mitochondria ultrastructure, Mitochondrial Membranes metabolism, Mitochondrial Membranes ultrastructure, Mitochondrial Membranes virology, Reactive Oxygen Species metabolism, Mitochondria metabolism, Parvoviridae Infections pathology, Parvoviridae Infections virology, Parvovirus, Canine physiology
- Abstract
Proper functioning of the mitochondria is crucial for the survival of the cell. Viruses are able to interfere with mitochondrial functions as they infect the host cell. Parvoviruses are known to induce apoptosis in infected cells, but the role of the mitochondria in parvovirus induced cytopathy is only partially known. Here we demonstrate with confocal and electron microscopy that canine parvovirus (CPV) associated with the mitochondrial outer membrane from the onset of infection. During viral entry a transient depolarization of the mitochondrial transmembrane potential and increase in ROS level was detected. Subsequently, mitochondrial homeostasis was normalized shortly, as detected by repolarization of the mitochondrial membrane and decrease of ROS. Indeed, activation of cell survival signalling through ERK1/2 cascade was observed early in CPV infected cells. At 12 hours post infection, concurrent with the expression of viral non-structural protein 1, damage to the mitochondrial structure and depolarization of its membrane were apparent. Results of this study provide additional insight of parvovirus pathology and also more general information of virus-mitochondria association.
- Published
- 2014
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32. High-fat feeding induces angiogenesis in skeletal muscle and activates angiogenic pathways in capillaries.
- Author
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Silvennoinen M, Rinnankoski-Tuikka R, Vuento M, Hulmi JJ, Torvinen S, Lehti M, Kivelä R, and Kainulainen H
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- Animals, Blood Glucose analysis, Blotting, Western, Capillaries physiology, Cytochromes c metabolism, Dietary Fats pharmacology, Immunohistochemistry, Male, Mice, Mice, Inbred C57BL, Mitochondria, Muscle enzymology, Muscle, Skeletal enzymology, Real-Time Polymerase Chain Reaction, Capillaries drug effects, Dietary Fats administration & dosage, Muscle, Skeletal blood supply, Neovascularization, Physiologic drug effects
- Abstract
High-fat diet (HFD) increases fatty acid oxidation in skeletal muscles. We hypothesized that this leads to increased oxygen demand and thus to increased capillarization. We determined the effects of high-fat diet on capillarization and angiogenic factors in skeletal muscles of mice that were either active or sedentary. Fifty-eight C57BL/6 J mice were divided into four groups: low-fat diet sedentary (LFS), low-fat diet active (LFA), high-fat diet sedentary (HFS), and high-fat diet active (HFA). The mice in active groups were housed in cages with running wheels and the sedentary mice were housed in similar cages without running wheels. After 19 weeks HFS, LFA and HFA had higher capillary density and capillary-to-fiber-ratio in quadriceps femoris muscles than LFS. Capillarization was similar in HFS and HFA. To reveal possible mechanisms of HFD induced angiogenesis, we measured protein and mRNA levels of angiogenic factors VEGF-A, HIF-1α, PGC-1α and ERRα. VEGF-A protein levels were higher in muscles of HFS, LFA and HFA compared to LFS. However, no significant differences were observed between HFA and HFS. Protein levels of HIF-1α, PGC-1α, and ERRα were similar in all groups. However, the mRNA expression of HIF-1α and VEGF-A was up-regulated in capillaries but not in muscle fibers of HFS. The sedentary and active mice groups had similar mRNA expression levels of angiogenesis regulators studied. We conclude that high-fat feeding induces angiogenesis in skeletal muscle and up-regulates the gene expression of HIF-1α and VEGF-A in capillaries.
- Published
- 2013
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33. [Update on current care guidelines: ovarian cancer].
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Leminen A, Auranen A, Bützow R, Hietanen S, Komulainen M, Kuoppala T, Mäenpää J, Puistola U, Vuento M, Vuorela P, and Yliskoski M
- Subjects
- Antibodies, Monoclonal, Humanized administration & dosage, Bevacizumab, Cisplatin administration & dosage, Combined Modality Therapy, Female, Humans, Neoplasm, Residual drug therapy, Neoplasm, Residual surgery, Practice Guidelines as Topic, Survival Analysis, Taxoids administration & dosage, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Ovarian Neoplasms drug therapy, Ovarian Neoplasms surgery
- Abstract
Ovarian cancer is the most lethal gynaecological cancer. It appears that seemingly ovarian or primary peritoneal carcinomas, in fact, originate from fimbriae. BRCA1/2 mutation carriers are recommended for the removal of ovaries and fimbriae, to reduce the risk of cancer. Treatment of epithelial ovarian cancer is based on the combination of surgery and chemotherapy. The residual tumour volume at the primary operation is the most important predictive factor of survival. The best response at the primary treatment is observed with combination chemotherapy with taxane and platinum. Adding bevacitzumab to first line chemotherapy may improve survival.
- Published
- 2012
34. The putative metal coordination motif in the endonuclease domain of human Parvovirus B19 NS1 is critical for NS1 induced S phase arrest and DNA damage.
- Author
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Kivovich V, Gilbert L, Vuento M, and Naides SJ
- Subjects
- Amino Acid Motifs, Animals, Apoptosis, DNA Mutational Analysis, Endonucleases physiology, Hep G2 Cells, Humans, Mutagenesis, Site-Directed, Spodoptera, Viral Nonstructural Proteins analysis, Viral Nonstructural Proteins physiology, Virus Replication, DNA Damage, Endonucleases chemistry, Parvovirus B19, Human physiology, S Phase Cell Cycle Checkpoints, Viral Nonstructural Proteins chemistry
- Abstract
The non-structural proteins (NS) of the parvovirus family are highly conserved multi-functional molecules that have been extensively characterized and shown to be integral to viral replication. Along with NTP-dependent helicase activity, these proteins carry within their sequences domains that allow them to bind DNA and act as nucleases in order to resolve the concatameric intermediates developed during viral replication. The parvovirus B19 NS1 protein contains sequence domains highly similar to those previously implicated in the above-described functions of NS proteins from adeno-associated virus (AAV), minute virus of mice (MVM) and other non-human parvoviruses. Previous studies have shown that transient transfection of B19 NS1 into human liver carcinoma (HepG2) cells initiates the intrinsic apoptotic cascade, ultimately resulting in cell death. In an effort to elucidate the mechanism of mammalian cell demise in the presence of B19 NS1, we undertook a mutagenesis analysis of the protein's endonuclease domain. Our studies have shown that, unlike wild-type NS1, which induces an accumulation of DNA damage, S phase arrest and apoptosis in HepG2 cells, disruptions in the metal coordination motif of the B19 NS1 protein reduce its ability to induce DNA damage and to trigger S phase arrest and subsequent apoptosis. These studies support our hypothesis that, in the absence of replicating B19 genomes, NS1-induced host cell DNA damage is responsible for apoptotic cell death observed in parvoviral infection of non-permissive mammalian cells.
- Published
- 2012
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35. Gene expression centroids that link with low intrinsic aerobic exercise capacity and complex disease risk.
- Author
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Kivelä R, Silvennoinen M, Lehti M, Rinnankoski-Tuikka R, Purhonen T, Ketola T, Pullinen K, Vuento M, Mutanen N, Sartor MA, Reunanen H, Koch LG, Britton SL, and Kainulainen H
- Subjects
- Animals, Disease Models, Animal, Energy Metabolism genetics, Female, Gene Expression Profiling, Gene Expression Regulation, Genetic Predisposition to Disease, Immunohistochemistry, Metabolic Diseases genetics, Mitochondria metabolism, Muscle, Skeletal cytology, Myosin Heavy Chains genetics, Oligonucleotide Array Sequence Analysis, Oxygen Consumption genetics, Rats, Risk Factors, Exercise Tolerance genetics, Metabolic Diseases etiology, Myosin Heavy Chains metabolism, Physical Conditioning, Animal
- Abstract
A strong link exists between low aerobic exercise capacity and complex metabolic diseases. To probe this linkage, we utilized rat models of low and high intrinsic aerobic endurance running capacity that differ also in the risk for metabolic syndrome. We investigated in skeletal muscle gene-phenotype relationships that connect aerobic endurance capacity with metabolic disease risk factors. The study compared 12 high capacity runners (HCRs) and 12 low capacity runners (LCRs) from generation 18 of selection that differed by 615% for maximal treadmill endurance running capacity. On average, LCRs were heavier and had increased blood glucose, insulin, and triglycerides compared with HCRs. HCRs were higher for resting metabolic rate, voluntary activity, serum high density lipoproteins, muscle capillarity, and mitochondrial area. Bioinformatic analysis of skeletal muscle gene expression data revealed that many genes up-regulated in HCRs were related to oxidative energy metabolism. Seven mean mRNA expression centroids, including oxidative phosphorylation and fatty acid metabolism, correlated significantly with several exercise capacity and disease risk phenotypes. These expression-phenotype correlations, together with diminished skeletal muscle capillarity and mitochondrial area in LCR rats, support the general hypothesis that an inherited intrinsic aerobic capacity can underlie disease risks.
- Published
- 2010
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36. Mechanisms of cell death in canine parvovirus-infected cells provide intuitive insights to developing nanotools for medicine.
- Author
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Nykky J, Tuusa JE, Kirjavainen S, Vuento M, and Gilbert L
- Subjects
- Animals, Apoptosis, Caspases metabolism, Cats, Cell Cycle, Cell Line, DNA Damage, DNA Fragmentation, Dogs, Flow Cytometry, Gene Expression, HeLa Cells, Humans, Membrane Potential, Mitochondrial, Microscopy, Electron, Transmission, Microscopy, Fluorescence, Nanomedicine, Necrosis, Oncolytic Virotherapy trends, Parvovirus, Canine genetics, Viral Nonstructural Proteins genetics, Cell Death, Oncolytic Virotherapy methods, Parvovirus, Canine physiology
- Abstract
Viruses have great potential as nanotools in medicine for gene transfer, targeted gene delivery, and oncolytic cancer virotherapy. Here we have studied cell death mechanisms of canine parvovirus (CPV) to increase the knowledge on the CPV life cycle in order to facilitate the development of better parvovirus vectors. Morphological studies of CPV-infected Norden laboratory feline kidney (NLFK) cells and canine fibroma cells (A72) displayed characteristic apoptotic events. Apoptosis was further confirmed by activation of caspases and cellular DNA damage. However, results from annexin V-propidium iodide (PI) labeling and membrane polarization assays indicated disruption of the plasma membrane uncommon to apoptosis. These results provide evidence that secondary necrosis followed apoptosis. In addition, two human cancer cell lines were found to be infected by CPV. This necrotic event over apoptotic cell death and infection in human cells provide insightful information when developing CPV as a nanotool for cancer treatments.
- Published
- 2010
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37. Parvovirus B19 genotype specific amino acid substitution in NS1 reduces the protein's cytotoxicity in culture.
- Author
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Kivovich V, Gilbert L, Vuento M, and Naides SJ
- Subjects
- Amino Acid Substitution, Flow Cytometry, Genotype, Hep G2 Cells, Humans, Structure-Activity Relationship, Viral Nonstructural Proteins chemistry, Apoptosis drug effects, Parvovirus B19, Human genetics, Parvovirus B19, Human metabolism, Viral Nonstructural Proteins genetics, Viral Nonstructural Proteins toxicity
- Abstract
A clinical association between idiopathic liver disease and parvovirus B19 infection has been observed. Fulminant liver failure, not associated with other liver-tropic viruses, has been attributed to B19 in numerous reports, suggesting a possible role for B19 components in the extensive hepatocyte cytotoxicity observed in this condition. A recent report by Abe and colleagues (Int J Med Sci. 2007;4:105-9) demonstrated a link between persistent parvovirus B19 genotype I and III infection and fulminant liver failure. The genetic analysis of isolates obtained from these patients demonstrated a conservation of key amino acids in the nonstructural protein 1 (NS1) of the disease-associated genotypes. In this report we examine a conserved residue identified by Abe and colleagues and show that substitution of isoleucine 181 for methionine, as occurs in B19 genotype II, results in the reduction of B19 NS1-induced cytotoxicity of liver cells. Our results support the hypothesis that in the setting of persistent B19 infection, direct B19 NS1-induced cytotoxicity may play a role in idiopathic fulminant liver failure.
- Published
- 2010
- Full Text
- View/download PDF
38. Phase coexistence in a triolein-phosphatidylcholine system. Implications for lysosomal membrane properties.
- Author
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Pakkanen KI, Duelund L, Vuento M, and Ipsen JH
- Subjects
- Calorimetry, Differential Scanning, Electron Spin Resonance Spectroscopy, Glycerides chemistry, Membrane Fluidity, Phase Transition, Transition Temperature, Water chemistry, Lipid Bilayers chemistry, Lysosomes chemistry, Phosphatidylcholines chemistry, Triolein chemistry
- Abstract
The effects of tri- and monoglycerides on phospholipid (POPC) membranes were studied using spectroscopical methods. Triolein was found to form two types of POPC-rich membranes, both with POPC or as a three-component system with monopalmitin. These two membrane types were determined as co-existing phases based on their spontaneous and stable separation and named heavy and light phase according to their sedimentation behaviour. Marked differences were seen in the physical properties of these phases, even though only minor compositional variation was detected. The light, less polar phase was found to be less ordered and more fluid and seemed to allow significantly lower amount of water penetration into the membrane-water interface than pure POPC membrane. The heavy phase, apart from their slightly altered water penetration, resembled more a pure POPC membrane. As triglycerides are present in lysosomal membranes, the present results can be seen as an implication for polarity-based water permeability barrier possibly contributing to the integrity of lysosomes.
- Published
- 2010
- Full Text
- View/download PDF
39. [Update on current care guidelines. Diagnosis, treatment and follow-up of cytological changes in the cervix, vagina and vulva].
- Author
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Pekka N, Anttila A, Bützow R, Heikkilä E, Hiltunen-Back E, Mäenpää J, Puistola U, Rantanen V, Rintala M, Räisänen I, Santalahti A, Talvensaari-Mattila A, Vartiainen J, Vuento M, and Yliskoski M
- Subjects
- Female, Finland epidemiology, Humans, Incidence, Mass Screening, Papanicolaou Test, Papillomavirus Infections diagnosis, Papillomavirus Infections therapy, Quality Control, Uterine Cervical Neoplasms pathology, Vaginal Smears, Cervix Uteri pathology, Practice Guidelines as Topic, Uterine Cervical Neoplasms diagnosis, Uterine Cervical Neoplasms therapy, Vagina pathology, Vulva pathology
- Abstract
Approximately 150 cervical cancer cases are diagnosed in Finland annually. Both incidence and mortality have decreased by 80% since organised screening began. Recently, screening based on primary HPV-testing with Pap-smear triage has been shown to be more sensitive and more specific among women over 35 years old in randomised studies and thus may be implemented in routine. Abnormal findings in Pap smears indicate management. Confirmed CIN1 lesions are followed up and CIN2 and worse lesions treated. Follow-up after treatment should be reliably arranged, because elevated risk of cancer remains over 20 years after treatment. Quality control is of utmost importance.
- Published
- 2010
40. Desipramine induces disorder in cholesterol-rich membranes: implications for viral trafficking.
- Author
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Pakkanen K, Salonen E, Mäkelä AR, Oker-Blom C, Vattulainen I, and Vuento M
- Subjects
- Animals, Antidepressive Agents, Tricyclic pharmacology, Cells, Cultured, Disease Models, Animal, Dogs, Molecular Structure, Cell Membrane drug effects, Cholesterol metabolism, Computer Simulation, Desipramine pharmacology, Parvovirus, Canine drug effects, Parvovirus, Canine physiology
- Abstract
In this study, the effect of desipramine (DMI) on phospholipid bilayers and parvoviral entry was elucidated. In atomistic molecular dynamics simulations, DMI was found to introduce disorder in cholesterol-rich phospholipid bilayers. This was manifested by a decrease in the deuterium order parameter S(CD) as well as an increase in the membrane area. Disordering of the membrane suggested DMI to destabilize cholesterol-rich membrane domains (rafts) in cellular conditions. To relate the raft disrupting ability of DMI with novel biological relevance, we studied the intracellular effect of DMI using canine parvovirus (CPV), a virus known to interact with endosomal membranes and sphingomyelin, as an intracellular probe. DMI was found to cause retention of the virus in intracellular vesicular structures leading to the inhibition of viral proliferation. This implies that DMI has a deleterious effect on the viral traffic. As recycling endosomes and the internal vesicles of multivesicular bodies are known to contain raft components, the effect of desipramine beyond the plasma membrane step could be caused by raft disruption leading to impaired endosomal function and possibly have direct influence on the penetration of the virus through an endosomal membrane.
- Published
- 2009
- Full Text
- View/download PDF
41. Early succession of bacterial biofilms in paper machines.
- Author
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Tiirola M, Lahtinen T, Vuento M, and Oker-Blom C
- Subjects
- Bacteria genetics, Biodiversity, Genes, rRNA, Molecular Sequence Data, Phylogeny, RNA, Bacterial genetics, RNA, Ribosomal, 16S genetics, Sequence Analysis, DNA, Bacteria classification, Bacteria growth & development, Biofilms growth & development, Environmental Microbiology, Industrial Microbiology, Paper
- Abstract
Formation of biofilms causes severe problems in paper machines, and hence financial costs. It would be preferable to prevent attachment of the primary-colonizing bacteria than to control the growth of secondary communities, which are sheltered by exopolysaccharide slime layers. We have therefore investigated the early succession of paper-machine biofilms by incubating stainless-steel test coupons in the process water-flow lines in two paper machines operating in slightly alkaline conditions in temperatures (45 and 49 degrees C) supporting thermophilic microbes. Microbial succession was profiled using length heterogeneity analysis of PCR-amplified 16S rRNA genes (LH-PCR) and linking the sequence data of the created 16S rRNA gene libraries to the dominant LH-PCR peaks. Although the bacterial fingerprints obtained from the attached surface communities varied slightly in different samples, the biomarker signals of the dominating primary-colonizing bacterial groups remained high over time in each paper machine. Most of the 16S rRNA gene copies in the early biofilms were assigned to the genera Rhodobacter, Tepidimonas, and Cloacibacterium. The dominance of these sequence types decreased in the developing biofilms. Finally, as phylogenetically identical primary-colonizers were detected in the two different paper mills, the machines evidently had similar environmental conditions for bacterial growth and potentially a common source of contamination.
- Published
- 2009
- Full Text
- View/download PDF
42. Method and apparatus using selected superparamagnetic labels for rapid quantification of immunochromatographic tests.
- Author
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Laitinen MP, Salmela J, Gilbert L, Kaivola R, Tikkala T, Oker-Blom C, Pekola J, and Vuento M
- Abstract
A rapid method and instrumentation for quantification of immunochromatographic tests (ICT) are described. The principle and performance of the method was demonstrated by measuring the levels of human chorionic gonadotropin (hCG) present in urine. The test format was a sandwich assay using two distinct monoclonal antibodies directed against hCG. The first anti-hCG antibody was labeled with superparamagnetic particles whereas the second was immobilized as a narrow detection zone on a porous membrane. The human urine sample was mixed with superparamagnetic particles coated with the first anti-hCG antibody, and the mixture was allowed to migrate past the detection zone containing the second anti-hCG antibody. Capillary forces facilitated migration of the immune complexes along the porous membrane. The amount of superparamagnetic particle-labelled monoclonal anti-hCG bound to the detection zone was directly proportional to the amount of hCG present in the sample as detected by measuring magnetization in the detector coil. The method had a practical detection limit of 20 U/l (54 nM) of hCG per 5 μl of human urine and a linear range of three decades from 20 U/l to 10 000 U/l. In addition, the analysis was completed within less than 10 minutes. Thus, the test format should be suitable for fast detection and monitoring of a large variety of clinically important parameters and analytes.
- Published
- 2009
- Full Text
- View/download PDF
43. Parvovirus capsid disorders cholesterol-rich membranes.
- Author
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Pakkanen K, Kirjavainen S, Mäkelä AR, Rintanen N, Oker-Blom C, Jalonen TO, and Vuento M
- Subjects
- 1,2-Dipalmitoylphosphatidylcholine chemistry, Hydrogen-Ion Concentration, Capsid chemistry, Capsid Proteins chemistry, Cholesterol chemistry, Membrane Fluidity, Membranes, Artificial, Parvovirus, Canine chemistry
- Abstract
In this study canine parvovirus, CPV, was found to induce disorder in DPPC:cholesterol membranes in acidic conditions. This acidicity-induced fluidizing effect is suggested to originate from the N-terminus of the viral capsid protein VP1. In accordance with the model membrane studies, a fluidizing effect was seen also in the endosomal membranes during CPV infection implying an important functional role of the fluidization in the endocytic entry of the virus.
- Published
- 2009
- Full Text
- View/download PDF
44. Late steps of parvoviral infection induce changes in cell morphology.
- Author
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Pakkanen K, Nykky J, and Vuento M
- Subjects
- Animals, Cats, Cell Line, Cell Surface Extensions chemistry, Dogs, Parvoviridae Infections virology, Cell Shape, Cell Surface Extensions virology, Dog Diseases virology, Parvoviridae Infections veterinary, Parvovirus, Canine physiology
- Abstract
Previously, virus-induced non-filopodial extensions have not been encountered in connection with viral infections. Here, we report emergence of long extensions protruding from Norden laboratory feline kidney (NLFK) and A72 (canine fibroma) cells infected with canine parvovirus for 72 h. These extensions significantly differ in length and number from those appearing in control cells. The most striking feature in the extensions is the length, reaching up to 130 microm, almost twice the average length of a healthy NLFK cell. In A72 cells, the extensions were even longer, up to 200 microm. The results presented here also suggest that the events leading to the growth of these extensions start earlier in infection and abnormal extension growth is detectable already at 24-h post-infection (p.i.). These extensions may have a vital role in the cell-to-cell transmission of the virus.
- Published
- 2008
- Full Text
- View/download PDF
45. Ultrasonographic assessment of weight of the myomatous uterus: a pilot study using a new combined geometrical formula.
- Author
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Rovio PH, Luukkaala T, Vuento M, Oksa S, Sundström H, and Heinonen PK
- Subjects
- Adult, Cervix Uteri diagnostic imaging, Cervix Uteri pathology, Female, Humans, Hysterectomy, Leiomyoma pathology, Middle Aged, Organ Size, Pilot Projects, Prospective Studies, Ultrasonography, Uterine Neoplasms pathology, Uterus pathology, Leiomyoma diagnostic imaging, Models, Biological, Uterine Neoplasms diagnostic imaging
- Abstract
Objective: To evaluate the accuracy of a formula combining the prolate ellipsoid (uterine corpus) and cylinder (uterine cervix) formulas in estimating the preoperative weight of the total uterus using a transvaginal ultrasound probe to obtain the uterine dimensions for the formulas., Study Design: Three dimensions of the uterine corpus (length, width and anteroposterior diameter) and cervical length and cervical anteroposterior diameter were preoperatively determined using a transvaginal ultrasound probe in 12 women with symptomatic leiomyomas scheduled to undergo hysterectomy. In two patients whose uteruses were the largest, part of the measurements had to be taken with a transabdominal ultrasound. Three investigators repeated all the rounds of measurements three times, producing in total 108 of findings (12 subjects x 3 investigators x 3 rounds of measurements). The geometric formula of prolate ellipsoid was compared to a formula combining the ellipsoid and cylinder formulas for accuracy in predicting overall uterine size (corpus and cervix) through correlation with hysterectomy specimens. The weight of the uterus in grams was directly derived from the volume of the uterus., Results: All measurements of the uterine corpus and cervix could be obtained preoperatively with a transvaginal ultrasound probe except in two patients who had the largest uteruses. The plain, traditional formula for the prolate ellipsoid overestimated the weight of the uterus and differences between the estimated and the true weight were statistically significant. The difference was not significant when the formula combining the formulas of the prolate ellipsoid and cylinder was used., Conclusion: The new formula combining the prolate ellipsoid and cylinder formulas is more accurate in predicting the true total weight of the uterus than the plain prolate ellipsoid formula. The transvaginal ultrasound probe proved useful in evaluating the dimensions of the uterine corpus and cervix.
- Published
- 2008
- Full Text
- View/download PDF
46. Sphingomyelin induces structural alteration in canine parvovirus capsid.
- Author
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Pakkanen K, Karttunen J, Virtanen S, and Vuento M
- Subjects
- Animals, Capsid metabolism, Capsid Proteins chemistry, Dogs, Parvovirus, Canine metabolism, Phosphatidylserines metabolism, Phospholipases A2 metabolism, Capsid chemistry, Parvovirus, Canine chemistry, Sphingomyelins metabolism
- Abstract
One of the essential steps in canine parvovirus (CPV) infection, the release from endosomal vesicles, is dominated by interactions between the virus capsid and the endosomal membranes. In this study, the effect of sphingomyelin and phosphatidyl serine on canine parvovirus capsid and on the phospholipase A(2) (PLA(2)) activity of CPV VP1 unique N-terminus was analyzed. Accordingly, a significant (P< or =0.05) shift of tryptophan fluorescence emission peak was detected at pH 5.5 in the presence of sphingomyelin, whereas at pH 7.4 a similar but minor shift was observed. This effect may relate to the exposure of VP1 N-terminus in acidic pH as well as to interactions between sphingomyelin and CPV. When the phenomenon was further characterized using circular dichroism spectroscopy, differences in CPV capsid CD spectra with and without sphingomyelin and phosphatidyl serine were detected, corresponding to data obtained with tryptophan fluorescence. However, when the enzymatic activity of CPV PLA(2) was tested in the presence of sphingomyelin, no significant effect in the function of the enzyme was detected. Thus, the structural changes observed with spectroscopic techniques appear not to manipulate the activity of CPV PLA(2), and may therefore implicate alternative interactions between CPV capsid and sphingomyelin.
- Published
- 2008
- Full Text
- View/download PDF
47. Ultrasonographic-guided pervaginal cul-de-sac cytology in the follow-up of ovarian carcinoma.
- Author
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Vuento M, Salmi T, Klemi P, and Grénman S
- Subjects
- Adult, Aged, Aged, 80 and over, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Female, Follow-Up Studies, Humans, Middle Aged, Neoplasm Recurrence, Local diagnostic imaging, Neoplasm Recurrence, Local pathology, Neoplasm Staging, Ovarian Neoplasms drug therapy, Prospective Studies, Ultrasonography methods, Biopsy, Needle methods, Ovarian Neoplasms diagnostic imaging, Ovarian Neoplasms pathology
- Abstract
Unlabelled: The aim of this study was to compare, prospectively, traditional pervaginal cul-de-sac aspiration cytology with an ultrasonographic-guided aspirate in the detection of residual or recurrent ovarian carcinoma., Patients and Methods: Fifty-one patients with ovarian carcinoma were monitored during chemotherapy (21 patients) or follow-up (30 patients) after first-line treatment. All patients underwent both traditional blind pervaginal cul-de-sac aspiration cytology and an ultrasonographic-guided pervaginal aspirate. The samples were classified as class 0 or insufficient when no mesothelial cells were detected in the aspirate. The results of cytological classification of the aspirates were compared with each other according to sampling order., Results: Samples were classified as class 0 in 56% when the traditional cul-de-sac aspiration was taken first, and in 73% when ultrasonographic-guided aspiration was taken first (p = 0.249, Fisher's exact test). The number of class 0 samples was smaller among those taken second than among those taken first (22 (44%) vs. 33 (65%), p = 0.046). Four recurrences were detected during the mean follow-up of six months (range 2-11 months) in 30 patients who were followed-up after the first-line treatment. In one case, a positive cul-de-sac cytology was the first and only early indication of recurrence., Conclusion: The use of ultrasonography did not improve the accuracy of the cul-de-sac aspiration. The greater amount of fluid in the cul-de-sac during the second sampling might contribute to achieving a better result.
- Published
- 2007
48. [Not Available].
- Author
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Nieminen P, Anttila A, Bützow R, Heikkilä E, Hiltunen-Back E, Puistola U, Rantanen V, Räisänen I, Santalahti A, Talvensaari-Mattila A, Vartianen J, Vuento M, and Yliskoski M
- Published
- 2007
49. Truncated forms of viral VP2 proteins fused to EGFP assemble into fluorescent parvovirus-like particles.
- Author
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Gilbert L, Toivola J, Välilehto O, Saloniemi T, Cunningham C, White D, Mäkelä AR, Korhonen E, Vuento M, and Oker-Blom C
- Abstract
Fluorescence correlation spectroscopy (FCS) monitors random movements of fluorescent molecules in solution, giving information about the number and the size of for example nano-particles. The canine parvovirus VP2 structural protein as well as N-terminal deletion mutants of VP2 (-14, -23, and -40 amino acids) were fused to the C-terminus of the enhanced green fluorescent protein (EGFP). The proteins were produced in insect cells, purified, and analyzed by western blotting, confocal and electron microscopy as well as FCS. The non-truncated form, EGFP-VP2, diffused with a hydrodynamic radius of 17 nm, whereas the fluorescent mutants truncated by 14, 23 and 40 amino acids showed hydrodynamic radii of 7, 20 and 14 nm, respectively. These results show that the non-truncated EGFP-VP2 fusion protein and the EGFP-VP2 constructs truncated by 23 and by as much as 40 amino acids were able to form virus-like particles (VLPs). The fluorescent VLP, harbouring VP2 truncated by 23 amino acids, showed a somewhat larger hydrodynamic radius compared to the non-truncated EGFP-VP2. In contrast, the construct containing EGFP-VP2 truncated by 14 amino acids was not able to assemble into VLP-resembling structures. Formation of capsid structures was confirmed by confocal and electron microscopy. The number of fluorescent fusion protein molecules present within the different VLPs was determined by FCS. In conclusion, FCS provides a novel strategy to analyze virus assembly and gives valuable structural information for strategic development of parvovirus-like particles.
- Published
- 2006
- Full Text
- View/download PDF
50. Diversity of bacteria contaminating paper machines.
- Author
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Lahtinen T, Kosonen M, Tiirola M, Vuento M, and Oker-Blom C
- Subjects
- Bacteria genetics, Equipment Contamination, Intercellular Signaling Peptides and Proteins, Organometallic Compounds, Peptides, Phylogeny, RNA, Ribosomal, 16S, Bacteria classification, Bacteria isolation & purification, Biodiversity, Paper
- Abstract
Formation of microbial biofilms and slimes is a general and serious problem in the operation of paper machines. Studies of microbial populations in paper machine-derived biofilms have been conducted using standard microbiological procedures; however, the bacterial genera present in this type of samples as well as their diversity are quite poorly known. Here, the bacterial diversity of 38 process water and 22 biofilm samples from four different Finnish paper machines were analyzed by length heterogeneity analysis of PCR-amplified 16S ribosomal DNA (LH-PCR). In addition, sequencing of the amplified 16S rRNA gene from 69 clones was conducted for characterization of the bacterial genera present in biofilm and slime samples. The LH-PCR profiles of both the free-living (process waters) and immobilized (biofilms) bacteria were diverse at all stages of the papermaking process. Out of the 69 sequenced clones, 44 belonged to alpha-Proteobacteria, most of which were close to the nitrogen-fixing root nodule genera Sinorhizobium, Rhizobium and Azorhizobium. Other clones were assigned to beta- and gamma-Proteobacteria and the phylum Bacteroidetes. In addition, eight of the clones were assigned to a yet uncultivated phylum, TM7. Finally, epifluorescence microscopy revealed that Gram-negative bacteria were predominant in both the biofilm (65%) and process water (54%) samples and a small coccoid cell morphology was most common in all samples. Together, our results show that the analysis of microbial samples from paper machines using modern molecular biology techniques adds valuable information and should, therefore, be useful as a more specific and sensitive microbiological method for the paper industry. This information could further be applied, e.g., in the development of more specific and environmental friendly antimicrobial agents for paper mills.
- Published
- 2006
- Full Text
- View/download PDF
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