| 1. |
- Alfredson, Håkan, et al.
(författare)
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cDNA-arrays and real-time quantitative PCR techniques in the investigation of chronic Achilles tendinosis.
- 2003
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Ingår i: Journal of Orthopaedic Research. - 0736-0266 .- 1554-527X. ; 21:6, s. 970-975
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Tidskriftsartikel (refereegranskat)abstract
- The aetiology and pathogenesis of chronic painful Achilles tendinosis are unknown. This investigation aimed to use cDNA arrays and real-time quantitative polymerase chain reaction (real-time PCR) technique to study tendinosis and control tissue samples. Five patients (females mean age 57.1+/-4.3 (years+/-SD)) with chronic painful Achilles tendinosis were included. From all patients, one biopsy was taken from the area with tendinosis and one from a clinically normal area (control) of the tendon. The tissue samples were immediately immersed in RNAlater and frozen at -80 degrees C until RNA extraction. Portions of pooled RNA from control and tendinosis sites, respectively, were transcribed to cDNA, radioactively labelled (32P), hybridized to cDNA expression arrays, and exposed to phosphoimager screens over night. Expressions of specific genes, shown to be regulated in the cDNA array analysis, were analyzed in the individual samples using real-time PCR. cDNA arrays showed that gene expressions for matrix-metalloproteinase-2 (MMP-2), fibronectin subunit B (FNRB), vascular endothelial growth factor (VEGF), and mitogen-activated protein kinase p38 (MAPKp38) were up-regulated, while matrix-metalloproteinase-3 (MMP-3) and decorin were down-regulated, in tendinosis tissue compared with control tissue. Using real-time PCR, 4/5 and 3/5 patients showed up-regulation of MMP-2 and FNRB mRNA, respectively. For decorin, VEGF, and MAPKp38, real-time PCR revealed a great variability among patients. Interestingly, the mRNAs for several cytokines and cytokine receptors were not regulated, indicating the absence of an inflammatory process in chronic painful Achilles tendinosis. In conclusion, cDNA-arrays and real-time PCR can be used to study differences in gene expression levels between tendinosis and control tendon tissue.
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| 2. |
- Andersson, Agneta, et al.
(författare)
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Aquatic ecosystems at risk for occurrence of pathogenic bacteria
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Annan publikation (övrigt vetenskapligt/konstnärligt)abstract
- Pathogenic bacteria occur naturally in aquatic systems. Co-existence of bacteria and protozoa has led to development of predation resistance strategies, which is suggested to serve as a driver for evolution of pathogenic bacteria. However, the ecological mechanisms for selection for different types of predation resistant and pathogenic bacteria are poorly known. To disentangle effects from nutrient availability and protozoan predation pressure on the occurrence of varying predation resistant bacterial genera, an enrichment-dilution experiment was performed where an aquatic bacterial community was exposed to protozoa. Operational taxonomical units, specific for three predation resistant bacterial genera were identified; Pseudomonas, Rickettsia and Mycobacterium. These genera are also known to harbor species that are potentially pathogenic to mammals. Rickettsia and Mycobacterium were promoted where protozoa were abundant and the predation pressure high, while Pseudomonas dominated the bacterial community at the highest nutrient level where the predation pressure on bacteria were low. Our study thus indicates that waters of all nutrient states can harbor pathogenic bacteria, but that bacteria with different ecological strategies occur depending on nutrient level and perturbation. The generative model approach presented here provide a possibility to integrate environmental data in prediction models of pathogens in complex environments.
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| 3. |
- Golovliov, Igor, 1958-, et al.
(författare)
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Long-Term Survival of Virulent Tularemia Pathogens outside a Host in Conditions That Mimic Natural Aquatic Environments
- 2021
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Ingår i: Applied and Environmental Microbiology. - : Elsevier. - 0099-2240 .- 1098-5336. ; 87:6, s. 1-11
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Tidskriftsartikel (refereegranskat)abstract
- Francisella tularensis, the causative agent of the zoonotic disease tularemia, can cause seasonal outbreaks of acute febrile illness in humans with disease peaks in late summer to autumn. Interestingly, its mechanisms for environmental persistence between outbreaks are poorly understood. One hypothesis is that F. tularensis forms biofilms in aquatic environments. We utilized two fully virulent wild-type strains: FSC200 (Francisella tularensis subsp. holarctica) and Schu S4 (Francisella tularensis subsp. tularensis) and three control strains, the attenuated live vaccine strain (LVS; F. tularensis subsp. holarctica), a Schu S4 DwbtI mutant that is documented to form biofilms, and the low-virulence strain U112 of the closely related species Francisella novicida. Strains were incubated in saline solution (0.9% NaCl) microcosms for 24 weeks at both 4°C and 20°C, whereupon viability and biofilm formation were measured. These temperatures were selected to approximate winter and summer temperatures of fresh water in Scandinavia, respectively. U112 and Schu S4 DwbtI formed biofilms, but F. tularensis strains FSC200 and Schu S4 and the LVS did not. All strains exhibited prolonged viability at 4°C compared to 20°C. U112 and FSC200 displayed remarkable long-term persistence at 4°C, with only 1- and 2-fold log reductions, respectively, of viable cells after 24weeks. Schu S4 exhibited lower survival, yielding no viable cells by week 20. At 24weeks, cells from FSC200, but not from Schu S4, were still fully virulent in mice. Taken together, these results demonstrate biofilm-independent, long-term survival of pathogenic F. tularensis subsp. holarctica in conditions that mimic overwinter survival in aquatic environments.
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| 4. |
- Hägglund, Moa, et al.
(författare)
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Accounting for bacterial overlap between raw water communities and contaminating sources improves the accuracy of signature-based microbial source tracking
- 2018
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Ingår i: Frontiers in Microbiology. - : Frontiers Media S.A.. - 1664-302X. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- Microbial source tracking (MST) analysis is essential to identifying and mitigating the fecal pollution of water resources. The signature-based MST method uses a library of sequences to identify contaminants based on operational taxonomic units (OTUs) that are unique to a certain source. However, no clear guidelines for how to incorporate OTU overlap or natural variation in the raw water bacterial community into MST analyses exist. We investigated how the inclusion of bacterial overlap between sources in the library affects source prediction accuracy. To achieve this, large-scale sampling-including feces from seven species, raw sewage, and raw water samples from water treatment plants - was followed by 16S rRNA amplicon sequencing. The MST library was defined using three settings: (i) no raw water communities represented; (ii) raw water communities selected through clustering analysis; and (iii) local water communities collected across consecutive years. The results suggest that incorporating either the local background or representative bacterial composition improves MST analyses, as the results were positively correlated to measured levels of fecal indicator bacteria and the accuracy at which OTUs were assigned to the correct contamination source increased fourfold. Using the proportion of OTUs with high source origin probability, underpinning a contaminating signal, is a solid foundation in a framework for further deciphering and comparing contaminating signals derived in signature-based MST approaches. In conclusion, incorporating background bacterial composition of water in MST can improve mitigation efforts for minimizing the spread of pathogenic and antibiotic resistant bacteria into essential freshwater resources.
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| 5. |
- Kaden, Rene, et al.
(författare)
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Brucellosis outbreak in a Swedish kennel in 2013 : Determination of genetic markers for source tracing
- 2014
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Ingår i: Veterinary Microbiology. - : Elsevier BV. - 0378-1135 .- 1873-2542. ; 174:3–4, s. 523-530
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Tidskriftsartikel (refereegranskat)abstract
- Brucellosis is a highly infectious zoonotic disease but rare in Sweden. Nonetheless, an outbreak of canine brucellosis caused by an infected dog imported to Sweden was verified in 2013. In total 25 dogs were tested at least duplicated by the following approaches: real-time PCR for the detection of Brucella canis, a Brucella genus-specific real-time PCR, selective cultivation, and microscopic examination. The whole genome of B. canis strain SVA13 was analysed regarding genetic markers for epidemiological examination. The genome of an intact prophage of Roseobacter was detected in B. canis strain SVA13 with whole genome sequence prophage analysis (WGS-PA). It was shown that the prophage gene content in the American, African and European isolates differs remarkably from the Asian strains. The prophage sequences in Brucella may therefore serve of use as genetic markers in epidemiological investigations. Phage DNA fragments were also detected in clustered, regularly interspaced short palindromic repeats (CRISPR) in the genome of strain SVA13. In addition to the recommendations for genetic markers in Brucella outbreak tracing, our paper reports a validated two-step stand-alone real-time PCR for the detection of B. canis and its first successful use in an outbreak investigation.
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| 6. |
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| 7. |
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| 8. |
- Rehnstam, Ann-Sofi, 1959-, et al.
(författare)
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Blooms of sequence-specific culturable bacteria in the sea
- 1993
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Ingår i: FEMS Microbiology Letters. - 0378-1097 .- 1574-6968. ; 102:3-4, s. 161-166
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Tidskriftsartikel (refereegranskat)abstract
- Using specific deoxyoligonucleotide probes we have discovered seasonally strong (up to ∼ 100%) dominance of bacteria hybridizing to a single probe, in near shore waters off Scripps pier (32°53′N; 117°15′W). The probes were designed from partially sequenced 16S rRNA (V3 domain) of isolated marine bacteria. The results indicate that this approach may be used for studies of bacterial populations in the marine environment. We have shown that a number of genotypes that at times are dominant in the natural assemblages are culturable (and not, ‘viable-but-unculturable’). Additionally, our data suggests that the discrepancy between viable counts and direct counts in sea water samples can be explained by low plating efficiency.
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| 10. |
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