1. |
- Cruz, Javier, 1990-, et al.
(author)
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Stable 3D Inertial Focusing by High Aspect Ratio Curved Microfluidics
- 2021
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In: Journal of Micromechanics and Microengineering. - : Institute of Physics Publishing (IOPP). - 0960-1317 .- 1361-6439. ; 31
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Journal article (peer-reviewed)abstract
- Fine manipulation of particles is essential for the analysis of complex samples such as blood or environmental water, where rare particles of interest may be masked by millions of others. Inertial focusing is amongst the most promising techniques for this task, enabling label-free manipulation of particles with sub-micron resolution at very high flow rates. However, the phenomenon still remains difficult to predict due to the focus position shifting in tortuous ways as function of the channel geometry, flow rate and particle size. Here, we present a new line of microfluidics that exploit inertial focusing in High Aspect Ratio Curved (HARC) microchannels and overcome this limitation. Consisting of a single curved channel, HARC systems provide a highly predictable, single focus position near the centre of the inner wall, largely independent of the flow rate and particle size.An explanation of the mechanism of migration and focus of particles, together with its governing equations, is provided based on simulations in COMSOL Multiphysics and experimental results. HARC microchannels built in silicon-glass were used for experimental validation, achieving a high quality, single focus position for a range of microparticles with sizes of 0.7 - 1 µm and bacterial cells (Escherichia coli). The recovery of 1 µm particles was 99.84% with a factor four in concentration.With a stable focus position, we envision that HARC systems will bring the technology closer to implementation in laboratories for analysis of complex fluids with biological particles like cells and organelles.
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3. |
- Hou, Zining, et al.
(author)
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Time lapse investigation of antibiotic susceptibility using a microfluidic linear gradient 3D culture device
- 2014
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In: Lab on a Chip. - : Royal Society of Chemistry (RSC). - 1473-0197 .- 1473-0189. ; 14:17, s. 3409-3418
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Journal article (peer-reviewed)abstract
- This study reports a novel approach to quantitatively investigate the antibacterial effect of antibiotics on bacteria using a three-dimensional microfluidic culture device. In particular, our approach is suitable for studying the pharmacodynamics effects of antibiotics on bacterial cells temporally and with a continuous range of concentrations in a single experiment. The responses of bacterial cells to a linear concentration gradient of antibiotics were observed using time-lapse photography, by encapsulating bacterial cells in an agarose-based gel located in a commercially available microfluidics chamber. This approach generates dynamic information with high resolution, in a single operation, e. g., growth curves and antibiotic pharmacodynamics, in a well-controlled environment. No pre-labelling of the cells is needed and therefore any bacterial sample can be tested in this setup. It also provides static information comparable to that of standard techniques for measuring minimum inhibitory concentration (MIC). Five antibiotics with different mechanisms were analysed against wild-type Escherichia coli, Staphylococcus aureus and Salmonella Typhimurium. The entire process, including data analysis, took 2.5-4 h and from the same analysis, high-resolution growth curves were obtained. As a proof of principle, a pharmacodynamic model of streptomycin against Salmonella Typhimurium was built based on the maximal effect model, which agreed well with the experimental results. Our approach has the potential to be a simple and flexible solution to study responding behaviours of microbial cells under different selection pressures both temporally and in a range of concentrations.
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4. |
- Zining, Hou, et al.
(author)
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A microfluidic approach for dynamic investigation of the antibiotic susceptibility of bacteria
- 2013
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In: MME 2013 24th Micromechanics and Microsystems Europe Conference.
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Conference paper (peer-reviewed)abstract
- The possibility to study the effect on cell growth of a chemical is a valued tool for re- searchers in different areas, such as antibiotic resis- tance, cancer research and metabolic pathways. Traditional approaches need long time and no dynamic information is given. Microfluidics offers short diffusion lengths and steeper gradients for studies of antibiotic susceptibility, which could improve throughput greatly. By combining with time-lapse micrography, information on the dynam- ics may provide additional understanding. A micro- fluidic 3D cell culture chip was used to determine the MIC (minimum inhibitory concentration) and the dynamics of sub-MIC of E. coli against ampicillin and spectinomycin. And the same works have been done in Staphylococcus aureus and Salmonella. It is the first time to reveal this dynamic behaviour of bacteria against antibiotics quantitatively in a micro- fludic device. It is anticipated that it could be ex- tended to many other similar investigations in bio- logical and relevant pharmaceutical or clinical applications.
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5. |
- Andersson, Dan I, et al.
(author)
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Mechanisms and clinical relevance of bacterial heteroresistance
- 2019
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In: Nature Reviews Microbiology. - : Nature Publishing Group. - 1740-1526 .- 1740-1534. ; 17:8, s. 479-496
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Research review (peer-reviewed)abstract
- Antibiotic heteroresistance is a phenotype in which a bacterial isolate contains subpopulations of cells that show a substantial reduction in antibiotic susceptibility compared with the main population. Recent work indicates that heteroresistance is very common for several different bacterial species and antibiotic classes. The resistance phenotype is often unstable, and in the absence of antibiotic pressure it rapidly reverts to susceptibility. A common mechanistic explanation for the instability is the occurrence of genetically unstable tandem amplifications of genes that cause resistance. Due to their instability, low frequency and transient character, it is challenging to detect and study these subpopulations, which often leads to difficulties in unambiguously classifying bacteria as susceptible or resistant. Finally, in vitro experiments, mathematical modelling, animal infection models and clinical studies show that the resistant subpopulations can be enriched during antibiotic exposure, and increasing evidence suggests that heteroresistance can lead to treatment failure.
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6. |
- Arthurson, Veronica, et al.
(author)
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Effects on Glomus mosseae root colonization by Paenibacillus polymyxa and Paenibacillus brasilensis strains as related to soil P-availability in winter wheat
- 2011
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In: Applied and Environmental Soil Science. - : Hindawi Limited. - 1687-7667 .- 1687-7675.
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Journal article (peer-reviewed)abstract
- Greenhouse experiments were conducted to assess the effects of inoculating winter wheat (Triticum aestivum) with plant growth promoting rhizobacteria (PGPR) of the genusPaenibacillusunder phosphate P-limited soil conditions in the presence or absence of the arbuscular mycorrhizal fungus (AMF)Glomus mosseae. FourP. polymyxastrains and oneP. brasilensisstrain were compared at two cell concentrations (106and 108 cells g−1seeds) of inoculation, and surface sterilized AMF spores were added to pots. Mycorrhizal root colonization, plant growth, and plant uptake of phosphorus were analyzed. Bacterial phosphate solubilization was examined separatelyin vitro. MostP. polymyxastrains, isolated from wheat, had dramatic effectsper seon root growth and root P-content. No treatment gave significant effect on shoot growth. AMF root colonization levels and total plant uptake of P were much stimulated by the addition of mostP. polymyxastrains. The AM fungus alone and theP. brasilensis, alone or in combination with the fungus, did not affect total plant P-levels. Our results indicate that practical application of inoculation with plant host-specific rhizobacteria (i.e.,P. polymyxa) could positively influence uptake of phosphorus in P-deficient soils by wheat plants, provided that suitable AM fungi (e.g.,G. mosseae) are present.
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7. |
- Aydemir, Umut, et al.
(author)
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In situ assembly of an injectable cardiac stimulator
- 2024
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In: Nature Communications. - : NATURE PORTFOLIO. - 2041-1723. ; 15:1
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Journal article (peer-reviewed)abstract
- Without intervention, cardiac arrhythmias pose a risk of fatality. However, timely intervention can be challenging in environments where transporting a large, heavy defibrillator is impractical, or emergency surgery to implant cardiac stimulation devices is not feasible. Here, we introduce an injectable cardiac stimulator, a syringe loaded with a nanoparticle solution comprising a conductive polymer and a monomer that, upon injection, forms a conductive structure around the heart for cardiac stimulation. Following treatment, the electrode is cleared from the body, eliminating the need for surgical extraction. The mixture adheres to the beating heart in vivo without disrupting its normal rhythm. The electrofunctionalized injectable cardiac stimulator demonstrates a tissue-compatible Young's modulus of 21 kPa and a high conductivity of 55 S/cm. The injected electrode facilitates electrocardiogram measurements, regulates heartbeat in vivo, and rectifies arrhythmia. Conductive functionality is maintained for five consecutive days, and no toxicity is observed at the organism, organ, or cellular levels. Heart pacing devices are bulky or rely on surgery. Here, the authors present an injectable cardiac stimulator based on a nanoparticle solution which attaches to the heart and forms a conductive path to the skin for external connection. It can regulate heartbeats and is thereafter cleared from the body.
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9. |
- Biccler, Jorne Lionel, et al.
(author)
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Relapse Risk and Loss of Lifetime After Modern Combined Modality Treatment of Young Patients With Hodgkin Lymphoma : A Nordic Lymphoma Epidemiology Group Study
- 2019
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In: Journal of Clinical Oncology. - : AMER SOC CLINICAL ONCOLOGY. - 0732-183X .- 1527-7755. ; 37:9, s. 703-713
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Journal article (peer-reviewed)abstract
- PURPOSE: Estimates of short- and long-term survival for young patients with classic Hodgkin lymphoma (cHL) are of considerable interest. We investigated cHL prognosis in the era of contemporary treatment at different milestones during the follow-up.PATIENTS AND METHODS: On the basis of a Nordic cohort of 2,582 patients diagnosed at ages 18 to 49 years between 2000 and 2013, 5-year relapse risks and 5-year restricted losses in expectation of lifetime were estimated for all patients and for patients who achieved event-free survival (EFS) for 12 (EFS12), 24 (EFS24), 36 (EFS36) or 60 (EFS60) months. The median follow-up time was 9 years (range, 2.9 to 16.8 years).RESULTS: The 5-year overall survival was 95% (95% CI, 94% to 96%). The 5-year risk of relapse was 13.4% (95% CI, 12.1% to 14.8%) overall but decreased to 4.2% (95% CI, 3.8% to 4.6%) given that patients reached EFS24. Relapse risk for patients treated with six to eight courses of bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, and prednisone (BEACOPP) was comparable to that of patients treated with six to eight courses of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD) despite more adverse risk criteria among patients treated with BEACOPP. Both from diagnosis and if EFS24 was reached, the losses in expectation of lifetime during the following 5 years were small (from diagnosis, 45 days [95% CI, 35 to 54 days] and for patients who reached EFS24, 13 days [95% CI, 7 to 20 days]). In stage-stratified analyses of 5-year restricted loss in expectation of lifetime, patients with stages I to IIA disease had no noteworthy excess risk of death after they reached EFS24, whereas risk remained measurable for patients with stages IIB to IV cHL.CONCLUSION: Real-world data on young patients with cHL from the Nordic countries show excellent outcomes. The outlook is particularly favorable for patients who reach EFS24, which supports limited relapse-oriented clinical follow-up.
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10. |
- Blanco, Paula, et al.
(author)
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Antimicrobial Peptide Exposure Selects for Resistant and Fit Stenotrophomonas maltophilia Mutants That Show Cross-Resistance to Antibiotics
- 2020
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In: mSphere. - : AMER SOC MICROBIOLOGY. - 2379-5042. ; 5:5
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Journal article (peer-reviewed)abstract
- Antimicrobial peptides (AMPs) are essential components of the innate immune system and have been proposed as promising therapeutic agents against drug-resistant microbes. AMPs possess a rapid bactericidal mode of action and can interact with different targets, but bacteria can also avoid their effect through a variety of resistance mechanisms. Apart from hampering treatment by the AMP itself, or that by other antibiotics in the case of cross-resistance, AMP resistance might also confer cross-resistance to innate human peptides and impair the anti-infective capability of the human host. A better understanding of how resistance to AMPs is acquired and the genetic mechanisms involved is needed before using these compounds as therapeutic agents. Using experimental evolution and whole-genome sequencing, we determined the genetic causes and the effect of acquired de novo resistance to three different AMPs in the opportunistic pathogen Stenotrophomonas maltophilia, a bacterium that is intrinsically resistant to a wide range of antibiotics. Our results show that AMP exposure selects for high-level resistance, generally without any reduction in bacterial fitness, conferred by mutations in different genes encoding enzymes, transporters, transcriptional regulators, and other functions. Cross-resistance to AMPs and to other antibiotic classes not used for selection, as well as collateral sensitivity, was observed for many of the evolved populations. The relative ease by which high-level AMP resistance is acquired, combined with the occurrence of cross-resistance to conventional antibiotics and the maintained bacterial fitness of the analyzed mutants, highlights the need for careful studies of S. maltophilia resistance evolution to clinically valuable AMPs. IMPORTANCE Stenotrophomonas maltophilia is an increasingly relevant multidrug-resistant (MDR) bacterium found, for example, in people with cystic fibrosis and associated with other respiratory infections and underlying pathologies. The infections caused by this nosocomial pathogen are difficult to treat due to the intrinsic resistance of this bacterium against a broad number of antibiotics. Therefore, new treatment options are needed, and considering the growing interest in using AMPs as alternative therapeutic compounds and the restricted number of antibiotics active against S. maltophilia, we addressed the potential for development of AMP resistance, the genetic mechanisms involved, and the physiological effects that acquisition of AMP resistance has on this opportunistic pathogen.
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11. |
- Dahlin, Andreas P., et al.
(author)
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Methodological aspects on microdialysis protein sampling and quantification in biological fluids : an in vitro study on human ventricular CSF.
- 2010
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In: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 82:11, s. 4376-4385
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Journal article (peer-reviewed)abstract
- There is growing interest in sampling of protein biomarkers from the interstitial compartment of the brain and other organs using high molecular cutoff membrane microdialysis (MD) catheters. However, recent data suggest that protein sampling across such MD membranes is a highly complex process that needs to be further studied. Here, we report three major improvements for microdialysis sampling of proteins in complex biological matrixes. The improvements in this in vitro study using human ventricular cerebrospinal fluid as the sample matrix include increased fluid recovery control, decreased protein adsorption on the microdialysis membrane and materials, and novel quantitative mass spectrometry analysis. Dextrans in different concentrations and sizes were added to the perfusion fluid. It was found that dextrans with molecular mass 250 and 500 kDa provided a fluid recovery close to 100%. An improved fluid recovery precision could be obtained by self-assembly triblock polymer surface modification of the MD catheters. The modified catheters also delivered a significantly increased extraction efficiency for some of the investigated proteins. The final improvement was to analyze the dialysates with isobaric tagged (iTRAQ) proteomics, followed by tandem mass spectrometric analysis. By using this technique, 48 proteins could be quantified and analyzed with respect to their extraction efficiencies. The novel aspects of microdialysis protein sampling, detection, and quantification in biological fluids presented in this study should be considered as a first step toward better understanding and handling of the challenges associated with microdialysis sampling of proteins. The next step is to optimize the developed methodology in vivo.
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12. |
- Ekström, Karin M., et al.
(author)
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"Det blir många nej" : konsumtionens meningar och villkor för barnfamiljer med knapp ekonomi
- 2010
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Reports (other academic/artistic)abstract
- Under de senaste decennierna har konsumtion kommit att bli ett allt viktigare område för människor när det gäller att visa och uppfatta socioekonomisk status. Det finns även forskare (Bauman 1998) som hävdar att konsumtionen blivit viktigare än arbetet när det gäller att visa status. I ett överflödssamhälle som det svenska är det framförallt den symboliska och positionella konsumtionen som är betydelsefull, d.v.s. sådan konsumtion som syftar till att visa omgivningen hur man vill uppfattas. På ett förenklat sätt kan man säga att denna typ av konsumtion rymmer två olika drivkrafter, social emulering och social distinktion. Emulering innebär att konsumtionen syftar till att man vill uppfattas som de flesta, som majoriteten konsumenter. Distinktion innebär däremot att genom konsumtion visa att man är annorlunda än de flesta, att man är unik och sticker ut från mängden. Dessa två drivkrafter är centrala och betydelsefulla för vår kunskap om konsumtionens mening(ar). Forskningsmässigt har det huvudsakliga intresset varit inriktat mot distinktion, d.v.s. konsumtion som kan tolkas som nyare trender och som utmärker sig. Den andra drivkraften, den sociala emuleringen, den ibland kanske gråare och vardagligare konsumtionen har inte tilldragit sig samma intresse. Detta har bl.a. fått som konsekvens att det är konsumenter som ligger i framkanten av förändrade konsumtionsmönster som tilldrar sig intresset medan de som genom sin konsumtion strävar efter att uppfattas som majoriteten, som ”normala” och efterföljare till trendsättare inte studerats i samma omfattning.
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13. |
- Ekström, Karin M., et al.
(author)
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Families Navigating the Landscape of Consumption in the Swedish Welfare Society
- 2010
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In: Journal of Macromarketing. - : SAGE Publications. - 0276-1467 .- 1552-6534. ; 30:4, s. 366-374
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Journal article (peer-reviewed)abstract
- Swedish welfare policy has in several ways promoted the individual rather than the family as a unit. This is not a phenomenon happening only in Sweden, but one occurring in many other Western countries where individualization is strongly emphasized. Sweden is, however, interesting as an example of a country with deep collective roots regarding issues of welfare. The welfare state has been, and to a major part still is, the main provider of social services, a fact that has affected the Swedish family in certain ways. During the last decades, the Swedish welfare society has changed toward marketization implying that individuals and families have to take on more responsibility regarding their welfare. The aim of this article is to discuss the transformation of Swedish families as consumers reflecting upon the effects of the Swedish welfare society.
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14. |
- Ekström, Karin M., et al.
(author)
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Försakelser och förhandlingar : vardag för barnfamiljer med knapp ekonomi
- 2011
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Reports (other academic/artistic)abstract
- Under de senaste decennierna har konsumtion kommit att bli ett allt viktigare område för människor när det gäller att uttrycka och uppfatta socioekonomisk status. Det finns även forskare (Bauman 1998) som hävdar att konsumtion blivit viktigare än arbete när det gäller att visa status. I ett överflödssamhälle som det svenska är det framför allt den symboliska och positionella konsumtionen som är betydelsefull, d.v.s. sådan konsumtion som syftar till att visa omgivningen hur man vill uppfattas. På ett förenklat sätt kan man säga att denna typ av konsumtion rymmer två olika drivkrafter, social emulering och social distinktion. Emulering innebär att konsumtionen syftar till att man vill uppfattas som de flesta, som majoriteten konsumenter. Distinktion innebär däremot att genom konsumtion visa att man är annorlunda än de flesta, att man är unik och sticker ut från mängden.
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22. |
- Ettema, Thijs J. G., et al.
(author)
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Rolf Bernander (1956-2014) : pioneer of the archaeal cell cycle Obituary
- 2014
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In: Molecular Microbiology. - : John Wiley & Sons. - 0950-382X .- 1365-2958. ; 92:5, s. 903-909
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Journal article (pop. science, debate, etc.)abstract
- On 19 January 2014 Rolf (Roffe') Bernander passed away unexpectedly. Rolf was a dedicated scientist; his research aimed at unravelling the cell biology of the archaeal domain of life, especially cell cycle-related questions, but he also made important contributions in other areas of microbiology. Rolf had a professor position in the Molecular Evolution programme at Uppsala University, Sweden for about 8 years, and in January 2013 he became chair professor at the Department of Molecular Biosciences, The Wenner-Gren Institute at Stockholm University in Sweden. Rolf was an exceptional colleague and will be deeply missed by his family and friends, and the colleagues and co-workers that he leaves behind in the scientific community. He will be remembered for his endless enthusiasm for science, his analytical mind, and his quirky sense of humour.
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23. |
- Ge, Xueliang, et al.
(author)
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Inhibition of translation termination by small molecules targeting ribosomal release factors
- 2019
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In: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 9
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Journal article (peer-reviewed)abstract
- The bacterial ribosome is an important drug target for antibiotics that can inhibit different stages of protein synthesis. Among the various classes of compounds that impair translation there are, however, no known small-molecule inhibitors that specifically target ribosomal release factors (RFs). The class I RFs are essential for correct termination of translation and they differ considerably between bacteria and eukaryotes, making them potential targets for inhibiting bacterial protein synthesis. We carried out virtual screening of a large compound library against 3D structures of free and ribosome-bound RFs in order to search for small molecules that could potentially inhibit termination by binding to the RFs. Here, we report identification of two such compounds which are found both to bind free RFs in solution and to inhibit peptide release on the ribosome, without affecting peptide bond formation.
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24. |
- Granhall, Ulf, et al.
(author)
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Bacterial community diversity in paper mills processing recycled paper
- 2010
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In: Journal of Industrial Microbiology & Biotechnology. - : Oxford University Press (OUP). - 1367-5435 .- 1476-5535. ; 37:10, s. 1061-1069
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Journal article (peer-reviewed)abstract
- Paper mills processing recycled paper suffer from biofouling causing roblems both in the mill and final product. The total bacterial ommunity composition and identification of specific taxa in the process ater and biofilms at the stock preparation and paper machine areas in a ill with recycled paper pulp was described by using a DNA-based pproach. Process water in a similar mill was also analyzed to nvestigate if general trends can be found between mills and over time. acterial community profiles, analyzed by terminal-restriction fragment ength polymorphism (T-RFLP), in process water showed that the dominant eaks in the profiles were similar between the two mills, although the verall composition was unique for each mill. When comparing process ater and biofilm at different locations within one of the mills, we bserved a separation according to location and sample type, with the iofilm from the paper machine being most different. 16S rRNA gene clone ibraries were generated and 404 clones were screened by RFLP analysis. rouping of RFLP patterns confirmed that the biofilm from the paper achine was most different. A total of 99 clones representing all RFLP atterns were analyzed, resulting in sequences recovered from nine acterial phyla, including two candidate phyla. Bacteroidetes epresented 45% and Actinobacteria 23% of all the clones. Sequences with imilarity to organisms implicated in biofouling, like Chryseobacterium pp. and Brevundimonas spp., were recovered from all samples even though he mill had no process problems during sampling, suggesting that they re part of the natural paper mill community. Moreover, many sequences howed little homology to as yet uncultivated bacteria implying that aper mills are interesting for isolation of new organisms, as well as or bioprospecting.
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26. |
- Heidarian, Sheida, et al.
(author)
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High prevalence of heteroresistance in Staphylococcus aureus is caused by a multitude of mutations in core genes
- 2024
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In: PLoS biology. - : Public Library of Science (PLoS). - 1544-9173 .- 1545-7885. ; 22:1
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Journal article (peer-reviewed)abstract
- Heteroresistance (HR) is an enigmatic phenotype where, in a main population of susceptible cells, small subpopulations of resistant cells exist. This is a cause for concern, as this small subpopulation is difficult to detect by standard antibiotic susceptibility tests, and upon antibiotic exposure the resistant subpopulation may increase in frequency and potentially lead to treatment complications or failure. Here, we determined the prevalence and mechanisms of HR for 40 clinical Staphylococcus aureus isolates, against 6 clinically important antibiotics: daptomycin, gentamicin, linezolid, oxacillin, teicoplanin, and vancomycin. High frequencies of HR were observed for gentamicin (69.2%), oxacillin (27%), daptomycin (25.6%), and teicoplanin (15.4%) while none of the isolates showed HR toward linezolid or vancomycin. Point mutations in various chromosomal core genes, including those involved in membrane and peptidoglycan/teichoic acid biosynthesis and transport, tRNA charging, menaquinone and chorismite biosynthesis and cyclic-di-AMP biosynthesis, were the mechanisms responsible for generating the resistant subpopulations. This finding is in contrast to gram-negative bacteria, where increased copy number of bona fide resistance genes via tandem gene amplification is the most prevalent mechanism. This difference can be explained by the observation that S. aureus has a low content of resistance genes and absence of the repeat sequences that allow tandem gene amplification of these genes as compared to gram-negative species.
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27. |
- Hjort, Karin, et al.
(author)
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Antibiotic Minimal Selective Concentrations and Fitness Costs during Biofilm and Planktonic Growth
- 2022
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In: mBio. - : American Society for Microbiology. - 2161-2129 .- 2150-7511. ; 13:3
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Journal article (peer-reviewed)abstract
- The use and misuse of antibiotics have resulted in the selection of difficult-to-treat resistant bacteria. Two key parameters that influence the selection of resistant bacteria are the minimal selective concentration (MSC) and the fitness cost of resistance, both of which have been measured during planktonic growth in several studies. However, bacterial growth most often occurs in biofilms, and it is unclear if and how these parameters differ under these two growth conditions. To address this knowledge gap, we compared a selection of several types of antibiotic-resistant Escherichia coli mutants during planktonic and biofilm growth to determine the fitness costs and MSCs. Biofilm-forming Escherichia coli strains are commonly found in catheter-associated and recurrent urinary tract infections. Isogenic strains of a biofilm-forming E. coli strain, differing only in the resistance mechanisms and the fluorescent markers, were constructed, and susceptible and resistant bacteria were grown in head-to-head competitions at various concentrations of antibiotics under planktonic and biofilm conditions. Mutants with resistance to five different antibiotics were studied. The results show that during both planktonic and biofilm growth, selection for the resistant mutants occurred for all antibiotics at sub-MICs far below the MIC of the antibiotic. Even though differences were seen, the MSC values and the fitness costs did not differ systematically between planktonic and biofilm growth, implying that despite the different growth modes, the basic selection parameters are similar. These findings highlight the risk that resistant mutants may, similarly to planktonic growth, also be selected at sub-MICs of antibiotics in biofilms.IMPORTANCE Our understanding of how and where antibiotic resistance is selected in response to antibiotic exposure is still limited, and this is particularly true for selective processes when bacteria are growing in biofilms, arguably the most significant mode of growth of bacteria in human and animal infections as well as in other settings. In this study, we compared how different types of resistant E. coli strains were selected in response to antibiotic exposure during planktonic and biofilm growth. Determination of the minimal selective concentrations (MSCs) and fitness costs of resistance showed that they were comparable under these two different conditions, even though some differences were observed. Importantly, the MSCs were far below the MICs for all mutants under both planktonic and biofilm growth, emphasizing the significance of low antibiotic concentrations in driving the emergence and enrichment of resistant bacteria. Our understanding of how and where antibiotic resistance is selected in response to antibiotic exposure is still limited, and this is particularly true for selective processes when bacteria are growing in biofilms, arguably the most significant mode of growth of bacteria in human and animal infections as well as in other settings. In this study, we compared how different types of resistant E. coli strains were selected in response to antibiotic exposure during planktonic and biofilm growth.
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28. |
- Hjort, Karin, et al.
(author)
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Bacterial chitinase with phytopathogen control capacity from suppressive soil revealed by functional metagenomics
- 2014
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In: Applied Microbiology and Biotechnology. - : Springer Science and Business Media LLC. - 0175-7598 .- 1432-0614. ; 98:6, s. 2819-2828
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Journal article (peer-reviewed)abstract
- Plant disease caused by fungal pathogens results in vast crop damage globally. Microbial communities of soil that is suppressive to fungal crop disease provide a source for the identification of novel enzymes functioning as bioshields against plant pathogens. In this study, we targeted chitin-degrading enzymes of the uncultured bacterial community through a functional metagenomics approach, using a fosmid library of a suppressive soil metagenome. We identified a novel bacterial chitinase, Chi18H8, with antifungal activity against several important crop pathogens. Sequence analyses show that the chi18H8 gene encodes a 425-amino acid protein of 46 kDa with an N-terminal signal peptide, a catalytic domain with the conserved active site F175DGIDIDWE183, and a chitinase insertion domain. Chi18H8 was expressed (pGEX-6P-3 vector) in Escherichia coli and purified. Enzyme characterization shows that Chi18H8 has a prevalent chitobiosidase activity with a maximum activity at 35 °C at pH lower than 6, suggesting a role as exochitinase on native chitin. To our knowledge, Chi18H8 is the first chitinase isolated from a metagenome library obtained in pure form and which has the potential to be used as a candidate agent for controlling fungal crop diseases. Furthermore, Chi18H8 may also answer to the demand for novel chitin-degrading enzymes for a broad range of other industrial processes and medical purposes.
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30. |
- Hjort, Karin, et al.
(author)
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Chitinase genes revealed and compared in bacterial isolates, DNA extracts and a metagenomic library from a phytopathogen-suppressive soil
- 2010
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In: FEMS Microbiology Ecology. - United Kingdom : Wiley-Blackwell Publishing Ltd.. - 0168-6496 .- 1574-6941. ; 71:2, s. 197-207
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Journal article (peer-reviewed)abstract
- Soil that is suppressive to disease caused by fungal pathogens is an interesting source to target for novel chitinases that might be contributing towards disease suppression. In this study, we screened for chitinase genes, in a phytopathogen-suppressive soil in three ways: (1) from a metagenomic library constructed from microbial cells extracted from soil, (2) from directly extracted DNA and (3) from bacterial isolates with antifungal and chitinase activities. Terminal restriction fragment length polymorphism (T-RFLP) of chitinase genes revealed differences in amplified chitinase genes from the metagenomic library and the directly extracted DNA, but approximately 40% of the identified chitinase terminal restriction fragments (TRFs) were found in both sources. All of the chitinase TRFs from the isolates were matched to TRFs in the directly extracted DNA and the metagenomic library. The most abundant chitinase TRF in the soil DNA and the metagenomic library corresponded to the TRF103 of the isolate Streptomyces mutomycini and/or Streptomyces clavifer. There were good matches between T-RFLP profiles of chitinase gene fragments obtained from different sources of DNA. However, there were also differences in both the chitinase and the 16S rRNA gene T-RFLP patterns depending on the source of DNA, emphasizing the lack of complete coverage of the gene diversity by any of the approaches used.
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31. |
- Hjort, Karin, et al.
(author)
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Community structure of actively growing bacterial populations in plant pathogen suppressive soil.
- 2007
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In: Microbial Ecology. - : Springer Science and Business Media LLC. - 0095-3628 .- 1432-184X. ; 53:3, s. 399-413
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Journal article (peer-reviewed)abstract
- The bacterial community in soil was screened by using various molecular approaches for bacterial populations that were activated upon addition of different supplements. Plasmodiophora brassicae spores, chitin, sodium acetate, and cabbage plants were added to activate specific bacterial populations as an aid in screening for novel antagonists to plant pathogens. DNA from growing bacteria was specifically extracted from the soil by bromodeoxyuridine immunocapture. The captured DNA was fingerprinted by terminal restriction fragment length polymorphism (T-RFLP). The composition of the dominant bacterial community was also analyzed directly by T-RFLP and by denaturing gradient gel electrophoresis (DGGE). After chitin addition to the soil, some bacterial populations increased dramatically and became dominant both in the total and in the actively growing community. Some of the emerging bands on DGGE gels from chitin-amended soil were sequenced and found to be similar to known chitin-degrading genera such as Oerskovia, Kitasatospora, and Streptomyces species. Some of these sequences could be matched to specific terminal restriction fragments on the T-RFLP output. After addition of Plasmodiophora spores, an increase in specific Pseudomonads could be observed with Pseudomonas-specific primers for DGGE. These results demonstrate the utility of microbiomics, or a combination of molecular approaches, for investigating the composition of complex microbial communities in soil.
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32. |
- Hjort, Karin, et al.
(author)
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Diversity and reductive evolution of mitochondria among microbial eukaryotes
- 2010
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In: Philosophical Transactions of the Royal Society of London. Biological Sciences. - : The Royal Society. - 0962-8436 .- 1471-2970. ; 365:1541, s. 713-727
-
Research review (peer-reviewed)abstract
- All extant eukaryotes are now considered to possess mitochondria in one form or another. Many parasites or anaerobic protists have highly reduced versions of mitochondria, which have generally lost their genome and the capacity to generate ATP through oxidative phosphorylation. These organelles have been called hydrogenosomes, when they make hydrogen, or remnant mitochondria or mitosomes when their functions were cryptic. More recently, organelles with features blurring the distinction between mitochondria, hydrogenosomes and mitosomes have been identified. These organelles have retained a mitochondrial genome and include the mitochondrial-like organelle of Blastocystis and the hydrogenosome of the anaerobic ciliate Nyctotherus. Studying eukaryotic diversity from the perspective of their mitochondrial variants has yielded important insights into eukaryote molecular cell biology and evolution. These investigations are contributing to understanding the essential functions of mitochondria, defined in the broadest sense, and the limits to which reductive evolution can proceed while maintaining a viable organelle.
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33. |
- Hjort, Karin, et al.
(author)
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Dynamics of Extensive Drug Resistance Evolution of Mycobacterium tuberculosis in a Single Patient During 9 Years of Disease and Treatment
- 2020
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In: Journal of Infectious Diseases. - : Oxford University Press (OUP). - 0022-1899 .- 1537-6613. ; 225:6, s. 1011-1020
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Journal article (peer-reviewed)abstract
- Mycobacterium tuberculosis is one of the hardest to treat bacterial pathogens with a high capacity to develop antibiotic resistance by mutations. Here we have performed whole-genome sequencing of consecutive M. tuberculosis isolates obtained during 9 years from a patient with pulmonary tuberculosis. The infecting strain was isoniazid resistant and during treatment it stepwise accumulated resistance mutations to 8 additional antibiotics. Heteroresistance was common and subpopulations with up to 3 different resistance mutations to the same drug coexisted. Sweeps of different resistant clones dominated the population at different time points, always coupled to resistance mutations coinciding with changes in the treatment regimens. Resistance mutations were predominant and no hitch-hiking, compensatory, or virulence-increasing mutations were detected, showing that the dominant selection pressure was antibiotic treatment. The results highlight the dynamic nature of M. tuberculosis infection, population structure, and resistance evolution and the importance of rapid antibiotic susceptibility tests to battle this pathogen.
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34. |
- Hjort, Karin, 1965-
(author)
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The cell cycle of the hyperthermophilic archaeal genus Sulfolobus
- 2002
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Doctoral thesis (other academic/artistic)abstract
- The third domain of life, Archaea is one of the three main evolutionary lineages together with the Bacteria and the Eukarya domains. The archaea are, despite their prokaryotic cell organisation, more closely related to eukaryotes than to bacteria in terms of the informational pathways (DNA replication, transcription and translation). Organisms from the archaeal hyperthermophilic genus Sulfolobus thrives in a hot (80°C), acidic (pH 2-4) and sulphur-rich environment.In my thesis, I have used a variety of different approaches to study the Sulfolobus cell cycle. After dilution of a stationary phase cell culture with fresh medium, synchronous cell cycle progression was obtained. From the synchronised cell culture experiment we could conclude that the major cell cycle events (nucleoid segregation, cell division and chromosome replication) were tightly coupled to each other and to cellular mass increase. Inhibitors of the elongation stage of chromosome replication, and of cell division, as well as drugs arresting the cell cycle in the post-replicative phase, were found in an in vivo screening of a range of antibiotics. The cell cycle was found to be regulated such that the previous cell cycle step had to be successfully accomplished before the next could initiate, except for DNA replication which could occur without an intervening cell division event.The replication pattern of Sulfolobus solfataricus was analysed using a marker frequency assay. From the results, we were able to determine that a single origin is utilized in vivo, that the replication directionality is bidirectional, and also an approximate location of the replication origin within the genome.Intracellular virus production in vivo of SIRV2 (Sulfolobus islandicus rod-shaped virus2) in Sulfolobus islandicus was also analysed. The effects on the host cell were determined, including loss of cell viability, inhibited initiation of replication at virus infection and DNA degradation and loss of cell integrity at the time of virus release. Also, for the first time intracellular virus DNA was visualized with flow cytometry.
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35. |
- Hjort, Karin, et al.
(author)
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Unstable tandem gene amplification generates heteroresistance (variation in resistance within a population) to colistin in Salmonella enterica
- 2016
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In: Molecular Microbiology. - : Wiley. - 0950-382X .- 1365-2958. ; 102:2, s. 274-289
-
Journal article (peer-reviewed)abstract
- Heteroresistance, a phenomenon where subpopulations of a bacterial isolate exhibit different susceptibilities to an antibiotic, is a growing clinical problem where the underlying genetic mechanisms in most cases remain unknown. We isolated colistin resistant mutants in Escherichia coli and Salmonella enterica serovar Typhimurium at different concentrations of colistin. Genetic analysis showed that genetically stable pmrAB point mutations were responsible for colistin resistance during selection at high drug concentrations for both species and at low concentrations for E. coli. In contrast, for S. Typhimurium mutants selected at low colistin concentrations, amplification of different large chromosomal regions conferred a heteroresistant phenotype. All amplifications included the pmrD gene, which encodes a positive regulator that up-regulates proteins that modify lipid A, and as a result increase colistin resistance. Inactivation and over-expression of the pmrD gene prevented and conferred resistance, respectively, demonstrating that the PmrD protein is required and sufficient to confer resistance. The heteroresistance phenotype is explained by the variable gene dosage of pmrD in a population, where sub-populations with different copy number of the pmrD gene show different levels of colistin resistance. We propose that variability in gene copy number of resistance genes can explain the heteroresistance observed in clinically isolated pathogenic bacteria.
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36. |
- Hjort, Martin, et al.
(author)
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In situ assembly of bioresorbable organic bioelectronics in the brain
- 2023
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In: Nature Communications. - 2041-1723. ; 14:1
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Journal article (peer-reviewed)abstract
- Bioelectronics can potentially complement classical therapies in nonchronic treatments, such as immunotherapy and cancer. In addition to functionality, minimally invasive implantation methods and bioresorbable materials are central to nonchronic treatments. The latter avoids the need for surgical removal after disease relief. Self-organizing substrate-free organic electrodes meet these criteria and integrate seamlessly into dynamic biological systems in ways difficult for classical rigid solid-state electronics. Here we place bioresorbable electrodes with a brain-matched shear modulus-made from water-dispersed nanoparticles in the brain-in the targeted area using a capillary thinner than a human hair. Thereafter, we show that an optional auxiliary module grows dendrites from the installed conductive structure to seamlessly embed neurons and modify the electrode's volume properties. We demonstrate that these soft electrodes set off a controlled cellular response in the brain when relaying external stimuli and that the biocompatible materials show no tissue damage after bioresorption. These findings encourage further investigation of temporary organic bioelectronics for nonchronic treatments assembled in vivo. Temporary bioelectronics can complement classical therapies in non-chronic treatments. Here, the authors describe the minimally invasive implantation of bioresorbable electrodes in the brain that form in situ from water-dispersed nanoparticles and show no tissue damage after bioresorption.
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37. |
- Hjort, Marcus, et al.
(author)
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Increased Inflammatory Activity in Patients 3 Months after Myocardial Infarction with Nonobstructive Coronary Arteries
- 2019
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In: Clinical Chemistry. - : AMER ASSOC CLINICAL CHEMISTRY. - 0009-9147 .- 1530-8561. ; 65:8, s. 1023-1030
-
Journal article (peer-reviewed)abstract
- BACKGROUND: Around 5%-10% of patients with myocardial infarction (MI) present with nonobstructive coronary arteries (MINOCA). We aimed to assess pathophysiological mechanisms in MINOCA by extensively evaluating cardiovascular biomarkers in the stable phase after an event, comparing MINOCA patients with cardiovascular healthy controls and MI patients with obstructive coronary artery disease (MI-CAD).METHODS: Ninety-one biomarkers were measured with a proximity extension assay 3 months after MI in 97 MINOCA patients, 97 age-and sex-matched MI-CAD patients, and 98 controls. Lasso analyses (penalized logistic regression models) and adjusted multiple linear regression models were used for statistical analyses.RESULTS: In the Lasso analysis (MINOCA vs MI-CAD), 8 biomarkers provided discriminatory value: P-selectin glycoprotein ligand 1, C-X-C motif chemokine 1, TNF-related activation-induced cytokine, and pappalysin-1 (PAPPA) with increasing probabilities of MINOCA, and tissue-type plasminogen activator, B-type natriuretic peptide, myeloperoxidase, and interleukin-1 receptor antagonist protein with increasing probabilities of MI-CAD. Comparing MINOCA vs controls, 7 biomarkers provided discriminatory value: N-terminal pro-B-type natriuretic peptide, renin, NF-kappa-B essential modulator, PAPPA, interleukin-6, and soluble urokinase plasminogen activator surface receptor with increasing probabilities of MINOCA, and agouti-related protein with increasing probabilities of controls. Adjusted multiple linear regression analyses showed that group affiliation was associated with the concentrations of 7 of the 8 biomarkers in the comparison MINOCA vs MI-CAD and 5 of the 7 biomarkers in MINOCA vs controls.CONCLUSIONS: Three months after the MI, the biomarker concentrations indicated greater inflammatory activity in MINOCA patients than in both MI-CAD patients and healthy controls, and a varying degree of myocardial dysfunction among the 3 cohorts.
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38. |
- Hjort, Torbjörn, et al.
(author)
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Consumption and inequality
- 2010
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In: Consumer behavior - A Nordic perspective. - 9789144055770 ; 1, s. 341-358
-
Book chapter (other academic/artistic)
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39. |
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40. |
- Hjort, Torbjörn, et al.
(author)
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Hidden consumers in marketing - the neglect of consumers with scarce resources in affluent societies
- 2009
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In: Journal of Marketing Management. - 0267-257X. ; 25:7-8, s. 697-712
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Journal article (peer-reviewed)abstract
- Abstract Research in marketing has neglected economic scarcity in affluent societies, with a few exceptions. Many affluent states are today facing financial difficulties and a global belief in the market as self-regulating, and in de-regulation have led to a focus on consumer agency. This has also contributed to a widening gap regarding opportunities to consume in affluent societies. The purpose of this article is to bring attention to the importance of considering economic scarcity in affluent societies among marketers in studies on consumption by using theoretical concepts from welfare studies such as inclusion and exclusion, participation and inequality. Researchers who ignore the consequences of the lower strata in the income hierarchy disregard the complexity of consumption. It is argued that regardless of income, we are all consumers, but with different opportunities and abilities. The hegemony of free choice needs to be challenged.
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41. |
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42. |
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43. |
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44. |
- Jerlström-Hultqvist, Jon, et al.
(author)
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Hydrogenosomes in the diplomonad Spironucleus salmonicida
- 2013
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In: Nature Communications. - : Springer Science and Business Media LLC. - 2041-1723. ; 4, s. 2493-
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Journal article (peer-reviewed)abstract
- Acquisition of the mitochondrion is a key event in the evolution of the eukaryotic cell, but diversification of the organelle has occurred during eukaryotic evolution. One example of such mitochondria-related organelles (MROs) are hydrogenosomes, which produce ATP by substrate- level phosphorylation with hydrogen as a byproduct. The diplomonad parasite Giardia intestinalis harbours mitosomes, another type of MRO. Here we identify MROs in the salmon parasite Spironucleus salmonicida with similar protein import and Fe-S cluster assembly machineries as in Giardia mitosomes. We find that hydrogen production is prevalent in the diplomonad genus Spironucleus, and that S. salmonicida MROs contain enzymes characteristic of hydrogenosomes. Evolutionary analyses of known hydrogenosomal components indicate their presence in the diplomonad ancestor, and subsequent loss in Giardia. Our results suggest that hydrogenosomes are metabolic adaptations predating the split between parabasalids and diplomonads, which is deeper than the split between animals and fungi in the eukaryotic tree.
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45. |
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46. |
- Jorgensen, Rasmus Rask Kragh, et al.
(author)
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Machine Learning-Based Survival Prediction Models for Progression-Free and Overall Survival in Advanced-Stage Hodgkin Lymphoma
- 2024
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In: JCO Clinical Cancer Informatics. - : Lippincott Williams & Wilkins. - 2473-4276. ; 8
-
Journal article (peer-reviewed)abstract
- PurposePatients diagnosed with advanced-stage Hodgkin lymphoma (aHL) have historically been risk-stratified using the International Prognostic Score (IPS). This study investigated if a machine learning (ML) approach could outperform existing models when it comes to predicting overall survival (OS) and progression-free survival (PFS).Patients and MethodsThis study used patient data from the Danish National Lymphoma Register for model development (development cohort). The ML model was developed using stacking, which combines several predictive survival models (Cox proportional hazard, flexible parametric model, IPS, principal component, penalized regression) into a single model, and was compared with two versions of IPS (IPS-3 and IPS-7) and the newly developed aHL international prognostic index (A-HIPI). Internal model validation was performed using nested cross-validation, and external validation was performed using patient data from the Swedish Lymphoma Register and Cancer Registry of Norway (validation cohort).ResultsIn total, 707 and 760 patients with aHL were included in the development and validation cohorts, respectively. Examining model performance for OS in the development cohort, the concordance index (C-index) for the ML model, IPS-7, IPS-3, and A-HIPI was found to be 0.789, 0.608, 0.650, and 0.768, respectively. The corresponding estimates in the validation cohort were 0.749, 0.700, 0.663, and 0.741. For PFS, the ML model achieved the highest C-index in both cohorts (0.665 in the development cohort and 0.691 in the validation cohort). The time-varying AUCs for both the ML model and the A-HIPI were consistently higher in both cohorts compared with the IPS models within the first 5 years after diagnosis.ConclusionThe new prognostic model for aHL on the basis of ML techniques demonstrated a substantial improvement compared with the IPS models, but yielded a limited improvement in predictive performance compared with the A-HIPI.
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47. |
- Kubicek-Sutherland, Jessica Z., et al.
(author)
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Antimicrobial peptide exposure selects for Staphylococcus aureus resistance to human defence peptides
- 2017
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In: Journal of Antimicrobial Chemotherapy. - : OXFORD UNIV PRESS. - 0305-7453 .- 1460-2091. ; 72:1, s. 115-127
-
Journal article (peer-reviewed)abstract
- Background: The clinical development of antimicrobial peptides (AMPs) is currently under evaluation to combat the rapid increase in MDR bacterial pathogens. However, many AMPs closely resemble components of the human innate immune system and the ramifications of prolonged bacterial exposure to AMPs are not fully understood. Objectives: We show that in vitro serial passage of a clinical USA300 MRSA strain in a host-mimicking environment containing host-derived AMPs results in the selection of stable AMP resistance. Methods: Serial passage experimentswere conducted using steadily increasing concentrations of LL-37, PR-39 or wheat germ histones. WGS and proteomic analysis by MS were used to identify the molecular mechanism associated with increased tolerance of AMPs. AMP-resistant mutants were characterized by measuring in vitro fitness, AMP and antibiotic susceptibility, and virulence in a mouse model of sepsis. Results: AMP-resistant Staphylococcus aureus mutants often displayed little to no fitness cost and caused invasive disease in mice. Further, this phenotype coincided with diminished susceptibility to both clinically prescribed antibiotics and human defence peptides. Conclusions: These findings suggest that therapeutic use of AMPs could select for virulent mutants with crossresistance to human innate immunity as well as antibiotic therapy. Thus, therapeutic use of AMPs and the implications of cross-resistance need to be carefully monitored and evaluated.
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48. |
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49. |
- Loza, Einars, et al.
(author)
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Structure-activity relationship studies on the inhibition of the bacterial translation of novel Odilorhabdins analogues
- 2020
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In: Bioorganic & Medicinal Chemistry. - : PERGAMON-ELSEVIER SCIENCE LTD. - 0968-0896 .- 1464-3391. ; 28:11
-
Journal article (peer-reviewed)abstract
- A structure-activity relationship (SAR) study of NOSO-95179, a nonapeptide from the Odilorhabdin class of antibacterials, was performed by systematic variations of amino acids in positions 2 and 5 of the peptide. A series of non-proteinogenic amino acids was synthesized in high enantiomeric purity from Williams' chiral diphenyloxazinone by highly diastereoselective alkylation or by aldol-type reaction. NOSO-95179 analogues for SAR studies were prepared using solid-phase peptide synthesis. Inhibition of bacterial translation by each of the synthesized Odilorhabdin analogues was measured using an in vitro test. For the most efficient analogues, antibacterial efficacy was measured against two wild-type Enterobacteriaceae (Escherichia coli and Klebsiella pneumoniae) and against an efflux defective E. coli strain (Delta tolC) to evaluate the impact of efflux on the antibacterial activity.
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50. |
- Morales-Lopez, Alvaro, et al.
(author)
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Influence of surface characteristics of polypropylene on E. coli and S. aureus biofilms : From conventional to additive manufacturing of bioprocess equipment
- 2024
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In: Applied Materials Today. - : Elsevier BV. - 2352-9407 .- 2352-9415. ; 39
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Journal article (peer-reviewed)abstract
- The fast-progressing landscape of the bioprocessing industry emphasizes innovation and efficiency enhancement, propelled by the integration of advanced solutions. Additive manufacturing technologies, particularly laserbased powder bed fusion with polypropylene, are pivotal in this industrial metamorphosis. However, despite the substantial scientific effort in the field, a significant gap exists in comprehending the surface characteristics of new surfaces and their implications for bacterial attachment and biofilm formation. This arises, in part, due to the absence of comprehensive and universally applicable topographical characterization analysis specifically designed for additively manufactured-fabricated surfaces. Typically, researchers tend to rely on the commonly used roughness parameter, Sa, that primarily quantifies the average height variation across a surface. Addressing this limitation is crucial for understanding the connection between surface characteristics and bacterial attachment dynamics. Here, we propose an innovative approach using surface analysis including confocal microscopy, advanced roughness measurements, and multivariate statistical analysis to uncover the connections between bacterial attachment for Gram negative Escherichia coli and Gram positive Staphylococcus aureus in early biofilm formation with surfaces produced by standardized and additively manufactured techniques. Finally, we advocate for the adoption of a set of roughness parameters that specifically describe the dale region of the surfaces. By doing so, we intend to establish direct links between surface texture and bacterial adhesion, thus contributing significantly to the advancement of both bioprocessing and additive manufacturing research domains.
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