| 1. |
- Axelsson, Ida, et al.
(författare)
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Impact of storage time prior to cryopreservation on mechanical properties of aortic homografts
- 2024
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Ingår i: Cell and Tissue Banking. - : Springer Science and Business Media LLC. - 1389-9333 .- 1573-6814.
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Tidskriftsartikel (refereegranskat)abstract
- Optimal time spans in homograft procurement are still debatable among tissue banks and needs to be further investigated. Cell viability decreases at longer preparation intervals, but the effect on collagen and elastic fibers has not been investigated to the same extent. These fibers are of importance to the homograft elasticity and strength. The objective of this study was to analyze the mechanical properties of homograft tissue at different time spans in the procurement process. Ten aortic homografts were collected at the Tissue Bank in Lund. Twelve samples were obtained from each homograft, cryopreserved in groups of three after 2–4 days, 7–9 days, 28–30 days, and 60–62 days in antibiotic decontamination. Mechanical testing was performed with uniaxial tensile tests, calculating elastic modulus, yield stress and energy at yield stress. Two randomly selected samples were assessed with light microscopy. Procurement generated a total of 120 samples, with 30 samples in each time group. Elastic modulus and yield stress was significantly higher in samples cryopreserved after 2–4 days (2.7 MPa (2.5-5.0) and 0.78 MPa (0.68-1.0)) compared to 7–9 days (2.2 MPa (2.0-2.6) and 0.53 MPa (0.46–0.69)), p = 0.008 and 0.011 respectively. Light microscopy did not show any difference in collagen and elastin at different time spans. There was a significant decrease in elastic modulus and yield stress after 7 days of decontamination at 4 °C compared to 2–4 days. This could indicate some deterioration of elastin and collagen at longer decontamination intervals. Clinical significance of these findings remains to be clarified.
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| 2. |
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| 3. |
- Ding, Mei, et al.
(författare)
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Secretome screening reveals immunomodulating functions of IFNα-7, PAP and GDF-7 on regulatory T-cells
- 2021
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Ingår i: Scientific Reports. - : Springer Nature. - 2045-2322. ; , s. 16767-
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Tidskriftsartikel (refereegranskat)abstract
- Regulatory T cells (Tregs) are the key cells regulating peripheral autoreactive T lymphocytes. Tregs exert their function by suppressing effector T cells. Tregs have been shown to play essential roles in the control of a variety of physiological and pathological immune responses. However, Tregs are unstable and can lose the expression of FOXP3 and suppressive functions as a consequence of outer stimuli. Available literature suggests that secreted proteins regulate Treg functional states, such as differentiation, proliferation and suppressive function. Identification of secreted proteins that affect Treg cell function are highly interesting for both therapeutic and diagnostic purposes in either hyperactive or immunosuppressed populations. Here, we report a phenotypic screening of a human secretome library in human Treg cells utilising a high throughput flow cytometry technology. Screening a library of 575 secreted proteins allowed us to identify proteins stabilising or destabilising the Treg phenotype as suggested by changes in expression of Treg marker proteins FOXP3 and/or CTLA4. Four proteins including GDF-7, IL-10, PAP and IFNα-7 were identified as positive regulators that increased FOXP3 and/or CTLA4 expression. PAP is a phosphatase. A catalytic-dead version of the protein did not induce an increase in FOXP3 expression. Ten interferon proteins were identified as negative regulators that reduced the expression of both CTLA4 and FOXP3, without affecting cell viability. A transcriptomics analysis supported the differential effect on Tregs of IFNα-7 versus other IFNα proteins, indicating differences in JAK/STAT signaling. A conformational model experiment confirmed a tenfold reduction in IFNAR-mediated ISG transcription for IFNα-7 compared to IFNα-10. This further strengthened the theory of a shift in downstream messaging upon external stimulation. As a summary, we have identified four positive regulators of FOXP3 and/or CTLA4 expression. Further exploration of these Treg modulators and their method of action has the potential to aid the discovery of novel therapies for both autoimmune and infectious diseases as well as for cancer.
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| 4. |
- Isaksson, Ida-Maria, et al.
(författare)
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Methods for 17 beta-oestradiol administration to rats
- 2011
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Ingår i: Scandinavian Journal of Clinical and Laboratory Investigation. - : Informa Healthcare. - 0036-5513 .- 1502-7686. ; 71:7, s. 583-592
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Tidskriftsartikel (refereegranskat)abstract
- Several studies indicate that the beneficial or harmful effects of oestrogens in stroke are dose-dependent. Rats are amongst the most frequently used animals in these studies, which calls for thoroughly validated methods for administering 17 beta-oestradiol to rats. In an earlier study we characterised three different administration methods for 17 beta-oestradiol over 42 days. The present study assesses the concentrations in a short time perspective, with the addition of a novel peroral method. Female Sprague-Dawley rats were ovariectomised and administered 17 beta-oestradiol by subcutaneous injections, silastic capsules, pellets and orally (in the nut-cream Nutella (R)), respectively. One group received 17 beta-oestradiol by silastic capsules without previous washout time. Blood samples were obtained after 30 minutes, 1, 2, 4, 8, 12, 24, 48 and 168 hours and serum 17 beta-oestradiol (and oestrone sulphate in some samples) was subsequently analysed. For long-term characterisation, one group treated perorally was blood sampled after 2, 7, 14, 21, 28, 35 and 42 days. At sacrifice, uterine horns were weighed and subcutaneous tissue samples were taken for histological assessment. The pellets, silastic capsule and injection groups produced serum 17 beta-oestradiol concentrations that were initially several orders of magnitude higher than physiological levels, while the peroral groups had 17 beta-oestradiol levels that were within the physiological range during the entire experiment. The peroral method is a promising option for administering 17 beta-oestradiol if physiological levels or similarity to womens oral hormone therapy are desired. Uterine weights were found to be a very crude measure of oestrogen exposure.
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| 5. |
- Isaksson, Ida-Maria, et al.
(författare)
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Methods for 17β-oestradiol administration to rats.
- 2011
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Ingår i: Scandinavian Journal of Clinical and Laboratory Investigation. - London, United Kingdom : Informa Healthcare. - 0036-5513 .- 1502-7686. ; 71:7, s. 583-592
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Tidskriftsartikel (refereegranskat)abstract
- Several studies indicate that the beneficial or harmful effects of oestrogens in stroke are dose-dependent. Rats are amongst the most frequently used animals in these studies, which calls for thoroughly validated methods for administering 17β-oestradiol to rats. In an earlier study we characterised three different administration methods for 17β-oestradiol over 42 days. The present study assesses the concentrations in a short time perspective, with the addition of a novel peroral method. Female Sprague-Dawley rats were ovariectomised and administered 17β-oestradiol by subcutaneous injections, silastic capsules, pellets and orally (in the nut-cream Nutella(®)), respectively. One group received 17β-oestradiol by silastic capsules without previous washout time. Blood samples were obtained after 30 minutes, 1, 2, 4, 8, 12, 24, 48 and 168 hours and serum 17β-oestradiol (and oestrone sulphate in some samples) was subsequently analysed. For long-term characterisation, one group treated perorally was blood sampled after 2, 7, 14, 21, 28, 35 and 42 days. At sacrifice, uterine horns were weighed and subcutaneous tissue samples were taken for histological assessment. The pellets, silastic capsule and injection groups produced serum 17β-oestradiol concentrations that were initially several orders of magnitude higher than physiological levels, while the peroral groups had 17β-oestradiol levels that were within the physiological range during the entire experiment. The peroral method is a promising option for administering 17β-oestradiol if physiological levels or similarity to women's oral hormone therapy are desired. Uterine weights were found to be a very crude measure of oestrogen exposure.
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| 6. |
- Isaksson, Johan, et al.
(författare)
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Semantic segmentation of microscopic images of H&E stained prostatic tissue using CNN
- 2017
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Ingår i: 2017 International Joint Conference on Neural Networks, IJCNN 2017 - Proceedings. - 9781509061815 ; 2017-May, s. 1252-1256
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Konferensbidrag (refereegranskat)abstract
- There is a need for an automatic Gleason scoring system that can be used for prostate cancer diagnosis. Today the diagnoses are determined by pathologists manually, which is both a complex and a time-consuming task. To reduce the pathologists' workload, but also to reduce variations between different pathologists, an automatic classification system would be of great use. Some previous works have aimed for this, but still more work needs to be done. It is probable that such a tool would benefit from having access to individually segmented, pathologically relevant objects from the images. Therefore, we have developed an algorithm for semantic segmentation of the microscopic images of H&E stained prostate tissue into Background, Stroma, Epithelial Cytoplasm and Nuclei. This algorithm is based on deep learning, or more specifically a convolutional neural network. The network design is inspired by architectures that previously have been proved successful in different applications. It consists of a contracting and an expanding part, which are symmetrical. We have reached an accuracy of 80 %, as measured by the mean of the intersection over union, for segmentation into four classes. Previous works have only investigated nuclei segmentation, and our network performed similar but for the more challenging task of four class segmentation.
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| 7. |
- Isaksson, Sven, et al.
(författare)
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A Novel Method to Analyze Social Transmission in Chronologically Sequenced Assemblages, Implemented on Cultural Inheritance of the Art of Cooking
- 2015
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 10:5
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Tidskriftsartikel (refereegranskat)abstract
- Here we present an analytical technique for the measurement and evaluation of changes in chronologically sequenced assemblages. To illustrate the method, we studied the cultural evolution of European cooking as revealed in seven cook books dispersed over the past 800 years. We investigated if changes in the set of commonly used ingredients were mainly gradual or subject to fashion fluctuations. Applying our method to the data from the cook books revealed that overall, there is a clear continuity in cooking over the ages - cooking is knowledge that is passed down through generations, not something (re-) invented by each generation on its own. Looking at three main categories of ingredients separately (spices, animal products and vegetables), however, disclosed that all ingredients do not change according to the same pattern. While choice of animal products was very conservative, changing completely sequentially, changes in the choices of spices, but also of vegetables, were more unbounded. We hypothesize that this may be due a combination of fashion fluctuations and changes in availability due to contact with the Americas during our study time period. The presented method is also usable on other assemblage type data, and can thus be of utility for analyzing sequential archaeological data from the same area or other similarly organized material.
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| 8. |
- Lindenfors, Patrik, 1964-, et al.
(författare)
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An empirical study of cultural evolution : the development of European cookery from medieval to modern times
- 2015
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Ingår i: Cliodynamics. - : California Digital Library (CDL). - 2373-7530. ; 6:2, s. 115-129
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Tidskriftsartikel (refereegranskat)abstract
- We have carried out an empirical study of long-term change in European cookery to test if the development of this cultural phenomenon matches a general hypothesis about cultural evolution: that human cultural change is characterized by cumulativity. Data from seven cookery books, evenly spaced across time, the oldest one written in medieval times (~1200) and the most recent one dating from late modernity (1999), were compared. Ten recipes from each of three categories (‘poultry recipes’, ‘fish recipes,’ and ‘meat recipes’) were arbitrarily selected from each cookery book by selecting the first ten recipes in each category, and the numbers (per recipe) of steps, separate partial processes, methods, ingredients, semi-manufactured ingredients, compound semi-manufactured ingredients (defined as semi-manufactured ingredients containing no less than two raw products), and self-made semi-manufactured ingredients were counted. Regression analyses were used to quantitatively compare the cookery from different ages. We found a significant increase in the numbers (per recipe) of steps, separate partial processes, methods, ingredients, and semi-manufactured ingredients. These significant increases enabled us to identify the development of cookery as an example of the general trend of cumulativity in long-term cultural evolution. The number of self-made semi-manufactured ingredients per recipe, however, tended to decrease over time, which may reflect the cumulative characteristics of cultural evolution at the level of society, considering the accumulation of knowledge that is required to industrialize food production.
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| 9. |
- Olofsson, Jessica, 1975, et al.
(författare)
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Direct Access and Control of the Intracellular Solution Environment in Single Cells
- 2009
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 81:5, s. 1810-1818
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Tidskriftsartikel (refereegranskat)abstract
- Methods that can control and vary the solution environment around single cells are abundant. In contrast, methods that offer direct access to the intracellular proteome and genome in single cells with the control, flexibility, and convenience given by microfluidic methods are both scarce and in great demand. Here, we present such a method based on using a microfluidic device mounted on a programmable scanning stage and cells on-chip permeabilized by the pore-forming glycoside digitonin. We characterized the on-chip digitonin poration, as well as the solution exchange within cells. Intracellular solution exchange times vary with the dose of exposure to digitonin from less than a second to tens of seconds. Also, the degree of permeabilization obtained for cells treated with the same dose varies considerably, especially for low doses of digitonin exposure and low permeabilities. With the use of the presented setup, the degree of permeabilization can be measured during the permeabilization process, which allows for "on-line" optimization of the digitonin exposure time. Using this calibrated permeabilization method, we demonstrate the generation of intracellular oscillations, intracellular gradients, and the delivery of substrate to initiate enzymatic reactions in situ. This method holds the potential to screen and titrate intracellular receptors or enzymes or to generate intracellular oscillations, useful in the study of signaling pathways and oscillation decoding among other applications.
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| 10. |
- Olofsson, Jessica, 1975, et al.
(författare)
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Probing enzymatic activity inside single cells.
- 2013
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Ingår i: Analytical Chemistry. - : American Chemical Society (ACS). - 0003-2700 .- 1520-6882. ; 85:21, s. 10126-33
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Tidskriftsartikel (refereegranskat)abstract
- We report a novel approach for determining the enzymatic activity within a single suspended cell. Using a steady-state microfluidic delivery device and timed exposure to the pore-forming agent digitonin, we controlled the plasma membrane permeation of individual NG108-15 cells. Mildly permeabilized cells (∼100 pores) were exposed to a series of concentrations of fluorescein diphosphate (FDP), a fluorogenic alkaline phosphatase substrate, with and without levamisole, an alkaline phosphatase inhibitor. We generated quantitative estimates for intracellular enzyme activity and were able to construct both dose-response and dose-inhibition curves at the single-cell level, resulting in an apparent Michaelis contant Km of 15.3 μM ± 1.02 (mean ± standard error of the mean (SEM), n = 16) and an inhibition constant Ki of 0.59 mM ± 0.07 (mean ± SEM, n = 14). Enzymatic activity could be monitored just 40 s after permeabilization, and five point dose-inhibition curves could be obtained within 150 s. This rapid approach offers a new methodology for characterizing enzyme activity within single cells.
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