| 51. |
- Giacalone, Davide, et al.
(författare)
-
Health and quality of life in an aging population : food and beyond
- 2014
-
Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- In Europe, as in much of the Western world, the number of citizens aged 65 and over has grown at an unprecedented rate and is expected to account for over 30% of the total population by 2060. Coupled with a steady increase in life expectancy, this massive demographic change calls for a major effort to ensure quality of life in our older population. A thorough understanding of the elderly as food consumers, their nutritional needs, and their food perception and preferences is increasingly recognized as key areas for future research. Food perception change at a later age as a result of the psychophysiological changes that occur with aging, such as decreased appetite and chemosensory acuity. The latter generally decrease food intake and the pleasure that the elderly derive from their meals, making the identification of possible compensation strategies (e.g., flavor enhancement, textural changes, etc.) essential to food producers interested in developing products for this increasingly important segment. Promoting food satisfaction among the elderly is also paramount to ensuring adequate nutritional intake. This aspect has major public health implications, such as preventing malnutrition and sarcopenia, which are leading causes of decreased independence and lower quality of life. Additionally, the importance of social and psychological factors is increasingly recognized. Many conditions related to aging (e.g., tiredness, loneliness) may prevent elderly people from preparing and enjoying meals, calling for alternative vehicles – such as tailor-made distribution channels, social food preparation and eating situations – for promoting healthy eating. In this workshop, a range of international speakers with relevant professional experience will present their latest work. More generally, it is our intention with this workshop to raise awareness of how sensory and consumer research can contribute to promote well-being among the elderly, and ultimately to expand the number of healthy life years as we age.
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| 52. |
- Giagounidis, Aristoteles, et al.
(författare)
-
Outcomes in RBC transfusion-dependent patients with Low-/Intermediate-1-risk myelodysplastic syndromes with isolated deletion 5q treated with lenalidomide: a subset analysis from the MDS-004 study
- 2014
-
Ingår i: European Journal of Haematology. - : Wiley. - 1600-0609 .- 0902-4441. ; 93:5, s. 429-438
-
Tidskriftsartikel (refereegranskat)abstract
- ObjectiveA subset analysis of the randomised, phase 3, MDS-004 study to evaluate outcomes in patients with International Prognostic Scoring System (IPSS)-defined Low-/Intermediate (Int)-1-risk myelodysplastic syndromes (MDS) with isolated del(5q). MethodsPatients received lenalidomide 10mg/d (days 1-21; n=47) or 5mg/d (days 1-28; n=43) on 28-d cycles or placebo (n=45). From the placebo and lenalidomide 5mg groups, 84% and 58% of patients, respectively, crossed over to lenalidomide 5 or 10mg at 16wk, respectively. ResultsRates of red blood cell-transfusion independence (RBC-TI) 182d were higher in the lenalidomide 10mg (57.4%; P<0.0001) and 5mg (37.2%; P=0.0001) groups vs. placebo (2.2%). Cytogenetic response rates (major+minor responses) were 56.8% (P<0.0001), 23.1% (P=0.0299) and 0%, respectively. Two-year cumulative risk of acute myeloid leukaemia progression was 12.6%, 17.4% and 16.7% in the lenalidomide 10mg, 5mg, and placebo groups, respectively. In a 6-month landmark analysis, overall survival was longer in lenalidomide-treated patients with RBC-TI 182d vs. non-responders (P=0.0072). The most common grade 3-4 adverse event was myelosuppression. ConclusionsThese data support the clinical benefits and acceptable safety profile of lenalidomide in transfusion-dependent patients with IPSS-defined Low-/Int-1-risk MDS with isolated del(5q).
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| 53. |
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| 54. |
- Gräslund, Torbjörn, et al.
(författare)
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Production of a Thermostable DNA Polymerase by Site-Specific Cleavage of a Heat-Eluted Affinity Fusion Protein
- 1997
-
Ingår i: Protein Expression and Purification. - : Elsevier BV. - 1046-5928 .- 1096-0279. ; 9, s. 125-132
-
Tidskriftsartikel (refereegranskat)abstract
- A novel strategy is described for bacterial expression and affinity purification of a recombinant truncated version of the heat-stable DNA polymerase I fromThermus aquaticus.The DNA polymerase ([Delta]Taq) was produced as a fusion to a serum albumin binding affinity handle (ABP) derived from streptococcal protein G. Based on the thermostability of the [Delta]TaqDNA polymerase, affinity-purified ABP-[Delta]Taqcould be heat-eluted from HSA columns by incubation at 85ï¿œC. To produce free [Delta]TaqDNA polymerase, efficient site-specific cleavage of the affinity tag was performed using a recombinant coxsackievirus 3C protease (3Cpro), also produced as an ABP affinity fusion. Thus, an integrated strategy could be devised where both the cleaved ABP affinity tag and the protease fusion could be recovered after site-specific cleavage using HSA-affinity chromatography. The flow-through fraction contained essentially pure [Delta]TaqDNA polymerase with full enzymatic activity.
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| 55. |
- Grövdal, Michael, et al.
(författare)
-
Maintenance treatment with azacytidine for patients with high-risk myelodysplastic syndromes (MDS) or acute myeloid leukaemia following MDS in complete remission after induction chemotherapy
- 2010
-
Ingår i: British Journal of Haematology. - : Wiley. - 0007-1048 .- 1365-2141. ; 150:3, s. 293-302
-
Tidskriftsartikel (refereegranskat)abstract
- This prospective Phase II study is the first to assess the feasibility and efficacy of maintenance 5-azacytidine for older patients with high-risk myelodysplastic syndrome (MDS), chronic myelomonocytic leukaemia and MDS-acute myeloid leukaemia syndromes in complete remission (CR) after induction chemotherapy. Sixty patients were enrolled and treated by standard induction chemotherapy. Patients that reached CR started maintenance therapy with subcutaneous azacytidine, 5/28 d until relapse. Promoter-methylation status of CDKN2B (P15 ink4b), CDH1 and HIC1 was examined pre-induction, in CR and 6, 12 and 24 months post CR. Twenty-four (40%) patients achieved CR after induction chemotherapy and 23 started maintenance treatment with azacytidine. Median CR duration was 13.5 months, >24 months in 17% of the patients, and 18-30.5 months in the four patients with trisomy 8. CR duration was not associated with CDKN2B methylation status or karyotype. Median overall survival was 20 months. Hypermethylation of CDH1 was significantly associated with low CR rate, early relapse, and short overall survival (P = 0.003). 5-azacytidine treatment, at a dose of 60 mg/m(2) was well tolerated. Grade III-IV thrombocytopenia and neutropenia occurred after 9.5 and 30% of the cycles, respectively, while haemoglobin levels increased during treatment. 5-azacytidine treatment is safe, feasible and may be of benefit in a subset of patients.
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| 56. |
- Grövdal, Michael, et al.
(författare)
-
Negative effect of DNA hypermethylation on the outcome of intensive chemotherapy in older patients with high-risk myelodysplastic syndromes and acute myeloid leukemia following myelodysplastic syndrome
- 2007
-
Ingår i: Clinical Cancer Research. - 1078-0432 .- 1557-3265. ; 13:23, s. 7107-7112
-
Tidskriftsartikel (refereegranskat)abstract
- PURPOSE: Promoter hypermethylation of, for example, tumor-suppressor genes, is considered to be an important step in cancerogenesis and a negative risk factor for survival in patients with myelodysplastic syndromes (MDS); however, its role for response to therapy has not been determined. This study was designed to assess the effect of methylation status on the outcome of conventional induction chemotherapy. EXPERIMENTAL DESIGN: Sixty patients with high-risk MDS or acute myeloid leukemia following MDS were treated with standard doses of daunorubicin and 1-beta-d-arabinofuranosylcytosine. Standard prognostic variables and methylation status of the P15(ink4b) (P15), E-cadherin (CDH), and hypermethylated in cancer 1 (HIC) genes were analyzed before treatment. RESULTS: Forty percent of the patients achieved complete remission (CR). CR rate was lower in patients with high WBC counts (P = 0.03) and high CD34 expression on bone marrow cells (P = 0.02). Whereas P15 status alone was not significantly associated with CR rate (P = 0.25), no patient with hypermethylation of all three genes achieved CR (P = 0.03). Moreover, patients with CDH methylation showed a significantly lower CR rate (P = 0.008), and CDH methylation retained its prognostic value also in the multivariate analysis. Hypermethylation was associated with increased CD34 expression, but not with other known predictive factors for response, such as cytogenetic profile. CONCLUSIONS: We show for the first time a significant effect of methylation status on the outcome of conventional chemotherapy in high-risk MDS and acute myelogenous leukemia following MDS. Provided confirmed in an independent study, our results should be used as a basis for therapeutic decision-making in this patient group.
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| 57. |
- Gullberg, Maria, et al.
(författare)
-
A single coxsackievirus B2 capsid residue controls cytolysis and apoptosis in rhabdomyosarcoma cells.
- 2010
-
Ingår i: Journal of Virology. - 0022-538X .- 1098-5514. ; 84:12, s. 5868-5879
-
Tidskriftsartikel (refereegranskat)abstract
- Coxsackievirus B2 (CVB2), one of six human pathogens of the group B coxsackieviruses within the enterovirus genus of Picornaviridae, causes a wide spectrum of human diseases ranging from mild upper respiratory illnesses to myocarditis and meningitis. The CVB2 prototype strain Ohio-1 (CVB2O) was originally isolated from a patient with summer grippe in the 1950s. Later on, CVB2O was adapted to cytolytic replication in rhabdomyosarcoma (RD) cells. Here, we present analyses of the correlation between the adaptive mutations of this RD variant and the cytolytic infection in RD cells. Using reverse genetics, we identified a single amino acid change within the exposed region of the VP1 protein (glutamine to lysine at position 164) as the determinant for the acquired cytolytic trait. Moreover, this cytolytic virus induced apoptosis, including caspase activation and DNA degradation, in RD cells. These findings contribute to our understanding of the host cell adaptation process of CVB2O and provide a valuable tool for further studies of virus-host interactions.
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| 58. |
- Gullberg, Maria, et al.
(författare)
-
Characterization of a putative ancestor of coxsackievirus B5.
- 2010
-
Ingår i: Journal of Virology. - 0022-538X .- 1098-5514. ; 84, s. 9695-9708
-
Tidskriftsartikel (refereegranskat)abstract
- Like other RNA viruses, coxsackievirus B5 (CVB5) exists as circulating heterogeneous populations of genetic variants. In this study, we present the reconstruction and characterization of a probable ancestral virion of CVB5. Phylogenetic analyses based on capsid protein encoding regions (the VP1 gene of 41 clinical isolates and the entire P1 region of eight clinical isolates) of CVB5 revealed two major co-circulating lineages. Ancestral capsid sequences were inferred from sequences of these contemporary CVB5 isolates using maximum likelihood methods. By using Bayesian phylodynamic analysis, the inferred VP1 ancestral sequence was dated back to 1854 (1807-1898). In order to study the properties of the putative ancestral capsid, the entire ancestral P1 sequence was synthesized de novo and inserted into the replicative backbone of an infectious CVB5 cDNA clone. Characterization of the recombinant virus in cell culture showed that fully functional infectious virus particles were assembled and that these viruses displayed properties similar to those of modern isolates, in terms of receptor preferences, plaque phenotype, growth characteristics and cell tropism. This is the first report describing resurrection and characterization of a picornavirus with a putative ancestral capsid. Our approach, including phylogenetics-based reconstruction of viral predecessors, could serve as a starting point for experimental studies of viral evolution and might also provide an alternative strategy in the development of vaccines.
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| 59. |
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| 60. |
- Hall, Michael, et al.
(författare)
-
Structural basis for glutathione-mediated activation of the virulence regulatory protein PrfA in Listeria
- 2016
-
Ingår i: Proceedings of the National Academy of Sciences of the United States of America. - : Proceedings of the National Academy of Sciences. - 0027-8424 .- 1091-6490. ; 113:51, s. 14733-14738
-
Tidskriftsartikel (refereegranskat)abstract
- Infection by the human bacterial pathogen Listeria monocytogenes is mainly controlled by the positive regulatory factor A (PrfA), a member of the Crp/Fnr family of transcriptional activators. Published data suggest that PrfA requires the binding of a cofactor for full activity, and it was recently proposed that glutathione (GSH) could fulfill this function. Here we report the crystal structures of PrfA in complex with GSH and in complex with GSH and its cognate DNA, the hly operator PrfA box motif. These structures reveal the structural basis for a GSH-mediated allosteric mode of activation of PrfA in the cytosol of the host cell. The crystal structure of PrfAWT in complex only with DNA confirms that PrfAWT can adopt a DNA binding-compatible structure without binding the GSH activator molecule. By binding to PrfA in the cytosol of the host cell, GSH induces the correct fold of the HTH motifs, thus priming the PrfA protein for DNA interaction.
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| 61. |
- Holmberg, Anders, et al.
(författare)
-
Towards 10-nm Soft X-Ray Zone Plate Fabrication
- 2011
-
Konferensbidrag (refereegranskat)abstract
- In this paper the latest efforts to improve our nanofabrication process for soft x‐ray zone plates is presented. The resolving power, which is proportional to the smallest outermost zone width of the zone plate, is increased by introducing cold development of the electron beam resist that is used for the patterning. With this process we have fabricated Ni zone plates with 13‐nm outermost zone and shown potential for making 11‐nm half‐pitch lines in the electron beam resist. Maintaining the diffraction efficiency of the zone plate is a great concern when the outermost zone width is decreased. To resolve this problem we have developed the so‐called Ni‐Ge zone plate in which the zone plate is build up by Ni and Ge, resulting in an increase of the diffraction efficiency. In a proof‐of‐principle experiment with 25‐nm Ni‐Ge zone plates, we have shown a doubling of the diffraction efficiency. When combined with cold development, the Ni‐Ge process has been shown to work down to 16‐nm half‐pitch. It is plausible that further refinement of the process will make it possible to go to 10‐nm outermost zone widths.
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| 62. |
- Israelsson, Stina, et al.
(författare)
-
Cytolytic replication of echoviruses in colon cancer cell lines
- 2011
-
Ingår i: Virology Journal. - 1743-422X. ; 8
-
Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Colorectal cancer is one of the most common cancers in the world, killing nearly 50% of patients afflicted. Though progress is being made within surgery and other complementary treatments, there is still need for new and more effective treatments. Oncolytic virotherapy, meaning that a cancer is cured by viral infection, is a promising field for finding new and improved treatments. We have investigated the oncolytic potential of several low-pathogenic echoviruses with rare clinical occurrence. Echoviruses are members of the enterovirus genus within the family Picornaviridae.METHODS: Six colon cancer cell lines (CaCo-2, HT29, LoVo, SW480, SW620 and T84) were infected by the human enterovirus B species echovirus 12, 15, 17, 26 and 29, and cytopathic effects as well as viral replication efficacy were investigated. Infectivity was also tested in spheroids grown from HT29 cells.RESULTS: Echovirus 12, 17, 26 and 29 replicated efficiently in almost all cell lines and were considered highly cytolytic. The infectivity of these four viruses was further evaluated in artificial tumors (spheroids), where it was found that echovirus 12, 17 and 26 easily infected the spheroids.CONCLUSIONS: We have found that echovirus 12, 17 and 26 have potential as oncolytic agents against colon cancer, by comparing the cytolytic capacity of five low-pathogenic echoviruses in six colon cancer cell lines and in artificial tumors.
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| 63. |
- Israelsson, Stina, et al.
(författare)
-
Improved replication efficiency of echovirus 5 after transfection of colon cancer cells using an authentic 5' RNA genome end methodology
- 2014
-
Ingår i: Investigational new drugs. - : Springer Science and Business Media LLC. - 0167-6997 .- 1573-0646. ; 32:6, s. 1063-1070
-
Tidskriftsartikel (refereegranskat)abstract
- Oncolytic virotherapy is a promising novel form of cancer treatment, but the therapeutic efficiency needs improvement. A potential strategy to enhance the therapeutic effect of oncolytic viruses is to use infectious nucleic acid as therapeutic agent to initiate an oncolytic infection, without administrating infectious viral particles. Here we demonstrate improved viral replication activation efficiency when transfecting cells with 5’ end authentic in vitro transcribed enterovirus RNA as compared to genomic RNA with additional non-genomic 5’ nucleotides generated by conventional cloning methods. We used echovirus 5 (E5) as an oncolytoc model virus due to its ability to replicate in and completely destroy five out of six colon cancer cell lines and kill artificial colon cancer tumors (HT29 spheroids), as shown here. An E5 infectious cDNA clone including a hammerhead ribozyme sequence was used to generate in vitro transcripts with native 5’ genome ends. In HT29 cells, activation of virus replication is approximately 20-fold more efficient for virus genome transcripts with native 5’ genome ends compared to E5 transcripts generated from a standard cDNA clone. This replication advantage remains when viral progeny release starts by cellular lysis 22 h post transfection. Hence, a native 5’ genomic end improves infection activation efficacy of infectious nucleic acid, potentially enhancing its therapeutic effect when used for cancer treatment. The clone design with a hammerhead ribozyme is likely to be applicable to a variety of oncolytic positive sense RNA viruses for the purpose of improving the efficacy of oncolytic virotherapy.
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| 64. |
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| 65. |
- Israelsson, Stina, et al.
(författare)
-
Studies of Echovirus 5 interactions with the cell surface: Heparan sulfate mediates attachment to the host cell.
- 2010
-
Ingår i: Virus Research. - : Elsevier BV. - 0168-1702 .- 1872-7492. ; 151:2, s. 170-176
-
Tidskriftsartikel (refereegranskat)abstract
- Infections caused by Echovirus 5 (E5), an enterovirus of the Picornaviridae family, have been associated with fever, rashes and sporadic cases of aseptic meningitis. To elucidate the receptor usage of this virus, the significance of a previously proposed integrin binding arginine-glycine-aspartic acid (RGD) motif found in the VP3 capsid protein was investigated, as well as the capacity of E5 to interact with heparan sulfate on the cell surface. Using the prototype strain E5 Noyce (E5N), an E5N mutant where the aspartic acid of the RGD motif has been substituted to a glutamic acid and clinical E5 isolates, the RGD motif of VP3 was found to be non-essential and hence not involved in integrin receptor binding. However, E5N and clinical E5 isolates interact with heparan sulfate at the cell surface, as demonstrated by virus replication inhibition assays using heparin and heparinase III, and studies of E5 interactions at the cell surface measured by real-time PCR analysis. In conclusion, E5 utilizes heparan sulfate as a cellular receptor, but the RGD motif of VP3 is not essential for E5 infectivity.
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| 66. |
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| 67. |
- Johannesson, Per, et al.
(författare)
-
A novel and rapid method to quantify cytolytic replication of picornaviruses in cell culture
- 2005
-
Ingår i: Journal of virological methods. - : Elsevier BV. - 0166-0934. ; 130, s. 117-123
-
Tidskriftsartikel (refereegranskat)abstract
- Determining viral titers is a key issue in a wide variety of studies regarding different aspects of virology. The standard methods used for determining picornavirus titers are endpoint titration assay and plaque assay, both time consuming and laborious. The method described uses the tetrazolium salt MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide) that is reduced to formazane by cellular dehydrogenase, genes shown to be down-regulated during picornavirus infection. The amount formazane produced correlates with the viral titers obtained and can easily be measured using an ELISA plate reader. The colorimetric method has been evaluated using virus types from different genera of the Picornaviridae family. The MTT method reduces the time spent on determining the viral titers and still maintains a reliable accuracy.
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| 68. |
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| 69. |
- Johansson, Per-Ola, et al.
(författare)
-
Design and synthesis of potent inhibitors of plasmepsin I and II : x-ray crystal structure of inhibitor in complex with plasmepsin II
- 2005
-
Ingår i: Journal of Medicinal Chemistry. - : American Chemical Society (ACS). - 0022-2623 .- 1520-4804. ; 48:13, s. 4400-4409
-
Tidskriftsartikel (refereegranskat)abstract
- New and potent inhibitors of the malarial aspartic proteases plasmepsin (Plm) I and II, from the deadliest malaria parasite Plasmodium falciparum, have been synthesized utilizing Suzuki coupling reactions on previously synthesized bromobenzyloxy-substituted statine-like inhibitors. The enzyme inhibition activity has been improved up to eight times by identifying P1 substituents that effectively bind to the continuous S1-S3 crevice of Plasmepsin I and II. By replacement of the bromo atom in the P1 p-bromobenzyloxy-substituted inhibitors with different aryl substituents, several inhibitors exhibiting Ki values in the low nanomolar range for both Plm I and II have been identified. Some of these inhibitors are also effective in attenuating parasite growth in red blood cells, with the best inhibitors, compounds 2 and 4, displaying 70% and 83% inhibition, respectively, at a concentration of 5 μM. The design was partially guided by the X-ray crystal structure disclosed herein of the previously synthesized inhibitor 1 in complex with plasmepsin II. © 2005 American Chemical Society.
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| 70. |
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| 71. |
- Johansson, Susanne, et al.
(författare)
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Cell culture propagation and biochemical analysis of the Ljungan virus prototype strain
- 2004
-
Ingår i: Biochemical and Biophysical Research Communication. - : Elsevier BV. - 0006-291X. ; 317:4, s. 1023-1029
-
Tidskriftsartikel (refereegranskat)abstract
- Ljungan virus (LV) is proposed as a potentially important rodent harbored viral human pathogen. Little is known about the biophysical nature of the virus and despite being molecularly characterized, progress in epidemiological and basic biological studies of LV has been hampered by the lack of a robust and reliable cell culture propagation system. Here we report the first description of an efficient lytic multi-cycle cell culture propagation of the LV prototype strain (87-012). Biophysical analysis of gradient purified LV virions generated by this system identified mature infectious virions to possess a sedimentation coefficient of 160S and in agreement with previous molecular prediction, polyprotein analysis suggests that the native virion is composed of only three major structural proteins. The nucleotide composition of the complete genome of the LV cell culture adapted virus was determined and compared to that of the parental prototype LV. Numerous mutations were observed scattered throughout the viral genome and particularly in VP1. The development of this cell culture system for LV should open new avenues in the study of LV biology, structure, pathogenesis, and prevalence of natural infection in the wider community.
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| 72. |
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| 73. |
- Johansson, Susanne
(författare)
-
Genomic Organization and Capsid Architecture of Ljungan Virus : a Novel Member of the Picornaviridae
- 2002
-
Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Ljungan virus är ett virus som isolerades i Sverige i mitten på nittiotalet. Under perioden 1989-1992 avled flera svenska elitorienterare plötsligt i hjärtmuskelinflammation. Man misstänkte att orienterarna kunde ha utsatts för en vektorburen infektion eftersom de exponeras för djur som finns i skog och mark under träning och tävling. Det är sedan tidigare känt att sorkar är den naturliga reservoaren för ett annat virus (Puumala virus) som kan orsaka njurskada hos människor. Sorkantalet i vissa delar av Sverige varierar kraftigt från år till år i ett cykliskt förlopp. Man fann ett samband mellan antalet sorkar och förekomsten av hjärtmuskelinflammation, typ 1 diabetes och Guillain-Barre's syndrom vilket ledde till att ett tidigare okänt virus, Ljungan virus, kunde isoleras från sorkar. Detta virus är ett litet RNA-virus som tillhör familjen Picornavirus. Till denna familj hör också flera kända virus såsom många av våra vanligt förekommande förkylningsvirus, men också virus som kan orsaka svåra sjukdomar, till exemel poliovirus och mul- och klövsjukevirus. Ljungan virus är ett nyupptäckt virus och därför är kunskapen om viruset begränsad. För att öka vår förståelse om viruset så har arvsmassan för tre svenska isolat (87-012, 174F och 145SL) av Ljungan virus kartlagts (artikel IV). Denna studie visade att Ljungan virusets arvmassa har flera unika egenskaper. Släktskapstudier visade att Ljungan virus är endast avlägset släkt med redan kända picornavirus och viruset bör därför utgöra en egen undergrupp i familjen (artikel III). Med kunskap om Ljungan virusets arvsmassa så var det möjligt att visa att Ljungan virus förekommer även på andra ställen än i Sverige (artikel VI). I mitten på 60-talet isolerades ett virus, M1146, från sork som fångats i Oregon, USA. Baserat på egenskaper hos proteinhöljet (kapsiden) så antog man då att M1146 var ett picornavirus. Studier av arvsmassan för detta virus visade att M1146 är närmast besläktat med de svenska Ljungan virus isolaten och har samma unika egenskaper i sin arvsmassa (artikel VI). Dessa studier har varit möjliga eftersom vi tidigare har utvecklat en metod för att producera stora mängder av hela arvsmassan (artikel I och II). Slutligen har det proteinhölje som innesluter och skyddar Ljungan virusets arvsmassa studerats (artikel V). Dessa studier visade att kapsiden är uppbyggd av tre proteiner och inte fyra som hos de flesta picornavirus. Dessa studier underlättades av att en enkel och effektiv metod för att odla Ljungan virus i provröret har utvecklats (artikel V). Sammantaget så har dessa studier försett oss med nya kunskaper om Ljungan virus som möjliggör fortsatta studier av dess biologi och eventuella förmåga att orsaka sjukdom hos människor och djur.
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| 74. |
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| 75. |
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| 76. |
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| 77. |
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| 78. |
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| 79. |
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| 80. |
- Jonsson, Nina, et al.
(författare)
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A rapid and efficient method for studies of virus interaction at the host cell surface using enteroviruses and real-time PCR
- 2009
-
Ingår i: Virology Journal. - 1743-422X. ; 6:Article ID: 217
-
Tidskriftsartikel (refereegranskat)abstract
- Background: Measuring virus attachment to host cells is of great importance when trying to identify novel receptors. The presence of a usable receptor is a major determinant of viral host range and cell tropism. Furthermore, identification of appropriate receptors is central for the understanding of viral pathogenesis and gives possibilities to develop antiviral drugs. Attachment is presently measured using radiolabeled and subsequently gradient purified viruses. Traditional methods are expensive and time-consuming and not all viruses are stable during a purification procedure; hence there is room for improvement. Real-time PCR (RT-PCR) has become the standard method to detect and quantify virus infections, including enteroviruses, in clinical samples. For instance, primers directed to the highly conserved 5' untranslated region (5'UTR) of the enterovirus genome enable detection of a wide spectrum of enteroviruses. Here, we evaluate the capacity of the RT-PCR technology to study enterovirus host cell interactions at the cell surface and compare this novel implementation with an established assay using radiolabeled viruses. Results: Both purified and crude viral extracts of CVB5 generated comparable results in attachment studies when analyzed with RT-PCR. In addition, receptor binding studies regarding viruses with coxsackie- nd adenovirus receptor (CAR) and/or decay accelerating factor (DAF) affinity, further demonstrated the possibility to use RT-PCR to measure virus attachment to host cells. Furthermore, the RT-PCR technology and crude viral extracts was used to study attachment with low multiplicity of infection (0.05 x 10(-4)TCID(50)/cell) and low cell numbers (250), which implies the range of potential implementations of the presented technique. Conclusion: We have implemented the well-established RT-PCR technique to measure viral attachment to host cells with high accuracy and reproducibility, at low cost and with less effort than traditional methods. Furthermore, replacing traditional methods with RT-PCR offers the opportunity to use crude virus containing extracts to investigate attachment, which could be considered as a step towards viral attachment studies in a more natural state.
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| 81. |
- Jonsson, Nina, et al.
(författare)
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Aichi virus infection in elderly people in Sweden.
- 2012
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Ingår i: Archives of Virology. - : Springer Science and Business Media LLC. - 0304-8608 .- 1432-8798. ; 157:7, s. 1365-1369
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Tidskriftsartikel (refereegranskat)abstract
- Aichi virus (AiV), genus Kobuvirus, family Picornaviridae, is associated with gastroenteritis in humans. Previous studies have shown high seroprevalence but low incidence (0.9-4.1%) in clinical samples. We report here the first detection of AiV in Sweden. Two hundred twenty-one specimens from hospitalized patients with diarrhea, who were negative for other enteric viruses, were included in the study. AiV were detected in three specimens, all from elderly patients. Phylogenetic analysis revealed that the three Swedish isolates belonged to genotype A and were genetically closest to European and Asian strains of AiV.
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| 82. |
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| 83. |
- Jonsson, Nina, et al.
(författare)
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Efficient replication of recombinant Enterovirus B types, carrying different P1 genes in the coxsackievirus B5 replicative backbone
- 2015
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Ingår i: Virus genes. - : Springer Science and Business Media LLC. - 0920-8569 .- 1572-994X. ; 50:3, s. 351-357
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Tidskriftsartikel (refereegranskat)abstract
- Recombination is an important feature in theevolution of the Enterovirus genus. Phylogenetic studies ofenteroviruses have revealed that the capsid genomic region(P1) is type specific, while the parts of the genome codingfor the non-structural proteins (P2–P3) are species specific.Hence, the genome may be regarded as consisting of twomodules that evolve independently. In this study, it wasinvestigated whether the non-structural coding part of thegenome in one type could support replication of a virus witha P1 region from another type of the same species. A cas-sette vector (pCas) containing a full-length cDNA copy ofcoxsackievirus B5 (CVB5) was used as a replicative back-bone. The P1 region of pCas was replaced with the corre-sponding part from coxsackievirus B3Nancy(CVB3N),coxsackievirus B6Schmitt(CVB6S), and echovirus 7Wal-lace(E7W), all members of theEnterovirus Bspecies. Thereplication efficiency after transfection with clone-derivedin vitro transcribed RNA was studied and compared withthat of pCas. All the recombinant viruses replicated with similar efficiencies and showed threshold cycle (Ct) values,tissue culture infectivity dose 50 %, and plaque-forming unittiters comparable to viruses generated from the pCas con-struct. In addition to this, a clone without the P1 region wasalso constructed, and Western Blot and immunofluorescencestaining analysis showed that the viral genome could betranslated and replicated despite the lack of the structuralprotein-coding region. To conclude, the replicative back-bone of the CVB5 cassette vector supports replication ofintraspecies constructs with P1 regions derived from othermembers of theEnterovirus Bspecies. In addition to this,the replicative backbone can be both translated and repli-cated without the presence of a P1 region.
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| 84. |
- Jonsson, Nina, et al.
(författare)
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Real-time polymerase chain reaction as a rapid and efficient alternative to estimation of picornavirus titers by tissue culture infectious dose 50% or plaque forming units
- 2009
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Ingår i: Microbiology and immunology. - : Wiley. - 0385-5600 .- 1348-0421. ; 53:3, s. 149-154
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Tidskriftsartikel (refereegranskat)abstract
- Quantification of viral infectious units is traditionally measured by methods based on forming plaques in semisolid media (PFU) or endpoint dilution of a virus-containing solution (TCID(50)), methods that are laborious, time-consuming and take on average 3-7 days to carry out. Quantitative real-time PCR is an established method to quantify nucleic acids at high accuracy and reproducibility, routinely used for virus detection and identification. In the present study, a procedure was developed using a two-step real-time PCR and the SYBR Green detection method to study whether there are correlations between TCID(50)/ml, PFU/ml and Ct values generated by real-time PCR enabling rapid and efficient calculation of titer equivalents when working with viruses in the research laboratory. In addition, an external standard with known concentrations was included using in vitro transcribed viral RNA, thus allowing the calculation of the amount of RNA copies needed for various applications (i.e. per plaque or TCID(50)).The results show that there is a correlation between the three quantification methods covering a wide range of concentration of viruses. Furthermore, a general regression line between TCID(50) and Ct values was obtained for all viruses included in the study, which enabled recording titer equivalents using real-time PCR. Finally, by including an external standard, the amount of RNA genomes generating one TCID(50) or PFU for each enterovirus serotype included was determined.
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| 85. |
- Jonsson, Pär, et al.
(författare)
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A strategy for modelling dynamic responses in metabolic samples characterized by GC/MS
- 2006
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Ingår i: Metabolomics. - : Springer Science and Business Media LLC. - 1573-3882 .- 1573-3890. ; 2:3, s. 135-143
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Tidskriftsartikel (refereegranskat)abstract
- A multivariate strategy for studying the metabolic response over time in urinary GC/MS data is presented and exemplified by a study of drug-induced liver toxicity in the rat. The strategy includes the generation of representative data through hierarchical multivariate curve resolution (H-MCR), highlighting the importance of obtaining resolved metabolite profiles for quantification and identification of exogenous (drug related) and endogenous compounds (potential biomarkers) and for allowing reliable comparisons of multiple samples through multivariate projections. Batch modelling was used to monitor and characterize the normal (control) metabolic variation over time as well as to map the dynamic response of the drug treated animals in relation to the control. In this way treatment related metabolic responses over time could be detected and classified as being drug related or being potential biomarkers. In summary the proposed strategy uses the relatively high sensitivity and reproducibility of GC/MS in combination with efficient multivariate curve resolution and data analysis to discover individual markers of drug metabolism and drug toxicity. The presented results imply that the strategy can be of great value in drug toxicity studies for classifying metabolic markers in relation to their dynamic responses as well as for biomarker identification.
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| 86. |
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| 87. |
- Jonsson, Pär, et al.
(författare)
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Predictive metabolite profiling applying hierarchical multivariate curve resolution to GC-MS data : a potential tool for multi-parametric diagnosis
- 2006
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Ingår i: Journal of Proteome Research. - : American Chemical Society. - 1535-3893 .- 1535-3907. ; 5:6, s. 1407-1414
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Tidskriftsartikel (refereegranskat)abstract
- A method for predictive metabolite profiling based on resolution of GC-MS data followed by multivariate data analysis is presented and applied to three different biofluid data sets (rat urine, aspen leaf extracts, and human blood plasma). Hierarchical multivariate curve resolution (H-MCR) was used to simultaneously resolve the GC-MS data into pure profiles, describing the relative metabolite concentrations between samples, for multivariate analysis. Here, we present an extension of the H-MCR method allowing treatment of independent samples according to processing parameters estimated from a set of training samples. Predictions or inclusion of the new samples, based on their metabolite profiles, into an existing model could then be carried out, which is a requirement for a working application within, e.g., clinical diagnosis. Apart from allowing treatment and prediction of independent samples the proposed method also reduces the time for the curve resolution process since only a subset of representative samples have to be processed while the remaining samples can be treated according to the obtained processing parameters. The time required for resolving the 30 training samples in the rat urine example was approximately 13 h, while the treatment of the 30 test samples according to the training parameters required only approximately 30 s per sample (approximately 15 min in total). In addition, the presented results show that the suggested approach works for describing metabolic changes in different biofluids, indicating that this is a general approach for high-throughput predictive metabolite profiling, which could have important applications in areas such as plant functional genomics, drug toxicity, treatment efficacy and early disease diagnosis.
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| 88. |
- Jääskeläinen, Anne J, et al.
(författare)
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Evidence of ljungan virus specific antibodies in humans and rodents, Finland.
- 2013
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Ingår i: Journal of Medical Virology. - : Wiley. - 0146-6615 .- 1096-9071. ; 85:11, s. 2001-2008
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Tidskriftsartikel (refereegranskat)abstract
- Ljungan virus (LV, genus Parechovirus, family Picornaviridae) is considered currently to be a rodent-borne virus. Despite suggested human disease associations, its zoonotic potential remains unclear. To date, LV antibody prevalence in both humans and rodents has not been studied. In this study, two different LV immunofluorescence assays (LV IFAs) were developed with LV genotypes 1 (LV strain 87-012G) and 2 (LV strain 145SLG), and cross-neutralization and -reaction studies were carried out with LV strain 145SLG. Finally, a panel of 37 Finnish sera was screened for anti-LV antibodies using two different LV IFAs (LV 145SLG and LV 87-012G) and a neutralization (NT) assay (LV 145SLG), and 50 samples from Myodes glareolus by LV IFA (LV 145SLG). The LV seroprevalence study showed 38% and 18% positivity in humans and M. glareolus, respectively. LV IFAs and NT assays were compared, and the results were in good agreement. The data are the first evidence of humans and rodents coming into contact with LV in Finland. Additional studies are required in order to acquire a better understanding of the prevalence, epidemiological patterns and possible disease association of LV infections.
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| 89. |
- Karlsson, Beatrice, et al.
(författare)
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Quasispecies dynamics and molecular evolution of human norovirus capsid P region during chronic infection
- 2009
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Ingår i: Journal of General Virology. - : Microbiology Society. - 0022-1317 .- 1465-2099. ; 90:2, s. 432-441
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Tidskriftsartikel (refereegranskat)abstract
- In this novel study, we have for the first time identified evolutionarily conserved capsid residues in an individual chronically infected with norovirus (GGII.3). From 2000 to 2003, a total of 147 P1-1 and P2 capsid sequences were sequenced and investigated for evolutionarily conserved and functionally important residues by the evolutionary trace (ET) algorithm. The ET algorithm revealed more absolutely conserved residues (ACR) in the P1-1 domain (47/53, 88 %) as compared with the P2 domain (86/133, 64 %). The capsid P1-1 and P2 domains evolved in time-dependent manner, with a distinct break point observed between autumn/winter of year 2000 (isolates P1, P3 and P5) and spring to autumn of year 2001 (isolates P11, P13 and P15), which presumably coincided with a change of clinical symptoms. Furthermore, the ET analysis revealed a similar receptor-binding pattern as reported for Norwalk and VA387 strains, with the CS-4 and CS-5 patch (Norwalk strain) including residues 329 and 377 and residues 306 and 310, respectively, all being ACR in all partitions. Most interesting was that residues 343, 344, 345, 374, 390 and 391 of the proposed receptor A and B trisaccharide binding site (VA387 strain) within the P2 domain remained ACR in all partitions, presumably because there was no selective advantage to alter the histo blood group antigens (HBGA) receptor binding specificity. In conclusion, this study provides novel insights to the evolutionary process of norovirus during chronic infection.
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| 90. |
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| 91. |
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| 92. |
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| 93. |
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| 94. |
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| 95. |
- Kim, MC, et al.
(författare)
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Development of duck hepatitis A virus type 3 vaccine and its use to protect ducklings against infections
- 2009
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Ingår i: Vaccine. - : Elsevier BV. - 0264-410X .- 1873-2518. ; 27:8, s. 6688-6694
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Tidskriftsartikel (refereegranskat)abstract
- A variant type of duck hepatitis A virus (DHAV), DHAV-3 was recently discovered in South Korea and China. Sequence analyses verified that the variant is genetically or serologically different from the DHAV-1 and DHAV-2 types. Duck hepatitis had been reported in South Korea since 1985 and an attenuated DHAV-1 vaccine had efficiently prevented epidemics of DHAV-1 until 2002. Despite the DHAV-1 based vaccine in use the novel DHAV-3 circulating in South Korea remains to be a threat to duckling farming. To develop a live attenuated vaccine against DHAV-3, a representative isolate, AP-04203, was therefore attenuated by repeated passages in SPF chicken embryos 100 times. The 100th passaged virus, AP-04203P100, did not cause clinical sign and mortality in 1-day-old ducklings as well as reversion of virulence capacity. The ducklings vaccinated with AP-04203P100 virus (10(3.0) ELD(50)/0.2 ml) on 1-day-old age via the intramuscular injection were well protected from 2 days after challenge with pathogenic AP04203P1 virus via the intramuscular route. In addition, the vaccine candidate also exhibited complete protection against currently circulating pathogenic DHAV-3 isolates. In conclusion, we demonstrate that the live attenuated virus, AP-04203P100, is a promising vaccine candidate facilitating the prevention of duck hepatitis caused by DHAV-3 around East Asia including South Korea.
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| 96. |
- Kirdok, Emrah, et al.
(författare)
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Metagenomic analysis of Mesolithic chewed pitch reveals poor oral health among stone age individuals
- 2024
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Ingår i: Scientific Reports. - : Springer Nature. - 2045-2322. ; 13:1
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Tidskriftsartikel (refereegranskat)abstract
- Prehistoric chewed pitch has proven to be a useful source of ancient DNA, both from humans and their microbiomes. Here we present the metagenomic analysis of three pieces of chewed pitch from Huseby Klev, Sweden, that were dated to 9,890-9,540 before present. The metagenomic profile exposes a Mesolithic oral microbiome that includes opportunistic oral pathogens. We compared the data with healthy and dysbiotic microbiome datasets and we identified increased abundance of periodontitis-associated microbes. In addition, trained machine learning models predicted dysbiosis with 70-80% probability. Moreover, we identified DNA sequences from eukaryotic species such as red fox, hazelnut, red deer and apple. Our results indicate a case of poor oral health during the Scandinavian Mesolithic, and show that pitch pieces have the potential to provide information on material use, diet and oral health.
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| 97. |
- Knowles, Charles H., et al.
(författare)
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Gastrointestinal neuromuscular pathology: guidelines for histological techniques and reporting on behalf of the Gastro 2009 International Working Group
- 2009
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Ingår i: Acta Neuropathologica. - : Springer Science and Business Media LLC. - 1432-0533 .- 0001-6322. ; 118:2, s. 271-301
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Tidskriftsartikel (refereegranskat)abstract
- The term gastrointestinal neuromuscular disease describes a clinically heterogeneous group of disorders of children and adults in which symptoms are presumed or proven to arise as a result of neuromuscular, including interstitial cell of Cajal, dysfunction. Such disorders commonly have impaired motor activity, i.e. slowed or obstructed transit with radiological evidence of transient or persistent visceral dilatation. Whilst sensorimotor abnormalities have been demonstrated by a variety of methods in these conditions, standards for histopathological reporting remain relatively neglected. Significant differences in methodologies and expertise continue to confound the reliable delineation of normality and specificity of particular pathological changes for disease. Such issues require urgent clarification to standardize acquisition and handling of tissue specimens, interpretation of findings and make informed decisions on risk-benefit of full-thickness tissue biopsy of bowel or other diagnostic procedures. Such information will also allow increased certainty of diagnosis, facilitating factual discussion between patients and caregivers, as well as giving prognostic and therapeutic information. The following report, produced by an international working group, using established consensus methodology, presents proposed guidelines on histological techniques and reporting for adult and paediatric gastrointestinal neuromuscular pathology. The report addresses the main areas of histopathological practice as confronted by the pathologist, including suction rectal biopsy and full-thickness tissue obtained with diagnostic or therapeutic intent. For each, indications, safe acquisition of tissue, histological techniques, reporting and referral recommendations are presented.
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| 98. |
- Knowles, N J, et al.
(författare)
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Family - Picornaviridae
- 2012
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Ingår i: Virus Taxonomy. - San Diego - London : Elsevier. - 9780123846846 ; , s. 855-881
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Bokkapitel (refereegranskat)abstract
- This chapter focuses on Picornaviridae family whose member genuses includeEnterovirus, Cardiovirus, Aphthovirus, Hepatovirus, and Parechovirus. The virions of this family consist of a capsid with no envelope and surrounds a core of ssRNA. Hydrated native particles are 30 nm in diameter, but vary from 22 to 30 nm in electron micrographs due to drying and flattening during preparation. The virions contain one molecule of positive sense, ssRNA, and possess a single long ORF. The UTRs at both termini contain regions of secondary structure, which are essential to genome function. In addition to the major CPs, 1A, 1B, 1C and 1D, and 3B (VPg), small amounts of 1AB (VP0) are commonly seen in lieu of one or more copies of 1A and 1B. Protein 1A is small in hepatoviruses, and 1AB is uncleaved in avihepatoviruses, kobuviruses, parechoviruses, and a number of unclassified picornaviruses. Some picornaviruses carry a sphingosine-like molecule in a cavity located inside 1D, and protein 1A generally has a molecule of myristic acid covalently attached to the amino terminal glycine. The virion RNA is infectious and serves as both the genome and the viral mRNA. Infection is generally cytolytic, but persistent infections are common with some species and reported with others. Poliovirus infected cells undergo extensive vacuolation as membranes are reorganized into viral replication complexes.
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| 99. |
- Knutsen, Carl Henrik, et al.
(författare)
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Conceptual and Measurement Issues in Assessing Democratic Backsliding
- 2023
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- This paper addresses three interrelated questions. First, how strong is the evidence that democracy has declined globally over the last decade? Second, how should we best measure (change in) democracy? Third, given that much of the recent evidencefor global backsliding comes from measurement projects that rely on expert ratings, is there evidence that experts have become harsher judges of democratic quality in recent years? We begin our analysis with a discussion of how to conceptualize democracy and democratic backsliding, stressing that for contested concepts such as democracy, no one operationalization is likely to reign supreme. We then dissect the distinction between “subjective” and “objective” measures, examining how measurement error can affect even seemingly objective indicators, and highlight how subjectivity pervades all measurement enterprises. Next, focusing on V–Dem’s methodology, we show—through both theoretical considerations and empirical tests—that it is highly unlikely that time-varying expert biases drive recent declines in estimates of the state of global democracy. Finally we evaluate Little and Meng’s (2023) recent attempt to assess the prevailing case for global backsliding using “objective” measures. We demonstrate multiple issues that make their measurement strategy ill-suited to studying trends in global democracy.
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| 100. |
- Knutsen, Carl Henrik, et al.
(författare)
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Conceptual and Measurement Issues in Assessing Democratic Backsliding
- 2024
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Ingår i: PS-POLITICAL SCIENCE & POLITICS. - 1049-0965 .- 1537-5935.
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Tidskriftsartikel (refereegranskat)abstract
- During the past decade, analyses drawing on several democracy measures have shown a global trend of democratic retrenchment. While these democracy measures use radically different methodologies, most partially or fully rely on subjective judgments to produce estimates of the level of democracy within states. Such projects continuously grapple with balancing conceptual coverage with the potential for bias (Munck and Verkuilen 2002; Przeworski et al. 2000). Little and Meng (L&M) (2023) reintroduce this debate, arguing that "objective" measures of democracy show little evidence of recent global democratic backsliding.1 By extension, they posit that time-varying expert bias drives the appearance of democratic retrenchment in measures that incorporate expert judgments. In this article, we engage with (1) broader debates on democracy measurement and democratic backsliding, and (2) L&M's specific data and conclusions.
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