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  • 1.
    Carlsson, Martin
    et al.
    Linnaeus University, Faculty of Health and Life Sciences, Department of Medicine and Optometry. Kalmar County Hospital.
    Nilsson, Ingela
    Kalmar County Hospital.
    Brudin, Lars
    Kalmar County Hospital ; Linköping University Hospital.
    Von, Siv-Ping
    Kalmar County Hospital.
    Wanby, Pär
    Kalmar County Hospital.
    Erythrocyte fatty acid composition does not influence levels of free, bioavailable, and total 25-hydroxy vitamin D2017In: Scandinavian Journal of Clinical and Laboratory Investigation, ISSN 0036-5513, E-ISSN 1502-7686, Vol. 77, no 1, p. 45-52Article in journal (Refereed)
    Abstract [en]

    In vitro, mono- and polyunsaturated fatty acids (FAs) may decrease the binding affinity of vitamin D metabolites for vitamin D-binding protein, which in turn may influence their bioavailability. FAs incorporated as phospholipids in erythrocyte (ery-) cell membranes reflect dietary intake. The purpose of this study was to investigate ery-FA composition in relation to markers for vitamin D. In healthy females (age 22.6 +/- 2.0 years) total 25(OH)D was measured by LC-MS/MS (n=78), free 25(OH)D with ELISA (n=64 of 78), and bioavailable 25(OH)D was calculated. Analysis of ery-FA composition was by gas chromatography (n=56 of 78). A strong correlation between total 25(OH)D and free 25(OH)D was seen (r=.66, p<.001), and between total-25(OH)D and bioavailable 25(OH)D (r=.68, p<.001). No correlations between 25(OH)D fractions and specific fatty acids were found, and in particular, no associations with mono- and poly-unsaturated FA compositions. All 25(OH)D fractions were correlated with leptin (total 25(OH)D (r=-.33, p<.003); bioavailable 25(OH)D (r=-.47, p<.001); free 25(OH)D (r=-.44, p<.001). Associations were found between PTH and total 25(OH)D (r=-.35, p=.002) and weaker between bioavailable 25(OH)D (r=-.35, p=.040) and free 25(OH)D (r=-.28, p=.079). All fractions of 25(OH)D appear to correlate in a similar way to PTH, BMI and body fat (leptin). No association was found between ery-FA composition and free/bioavailable 25(OH)D. It is unlikely that FAs are a strong uncoupling factor of DBP-bound 25(OH)D.

  • 2.
    Leander, Ellinor
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Artidentifiering av mögelsvamp med MALDI-TOF MS2018Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Rapid and accurate species identification is crucial for successful treatment of fungal infections, especially among immunosuppressed patients. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is used routinely at clinical laboratories to identify characteristic protein patterns of bacteria and yeast by the interpretation of protein spectra in a database for accurate species identification. The hard cell wall of the mold and the heterogeneous growth with varying protein expression due to maturation, complicates identification with MALDI-TOF MS. The potential benefits of this method compared to microscopy as traditional method are shortened turn-around times, safer species identification of more species that is independent on subjective morphological assessment. The purpose of the study was to investigate whether MALDI-TOF MS could be adapted and used for the identification of molds in clinical routine diagnostics. Four reference strains (Aspergillus niger, A.fumigatus, A.terreus, A.flavus) and a clinical isolate (A.terreus) were examined. The preparation methods (I) complete formic acid extraction, (II) direct application and (III) suspension in distilled water were used for analysis of spores and frontmycelium from younger and older mold cultures. Two different masspektradatabases for species identification were compared; routine database BDAL and the specialized mold database, Filamentous Fungi Library. Also the collecting technique of mold prior to analysis with MALDI-TOF MS was evaluated. Sometimes, the species identification improved after extraction of mold cultures, while in other cases direct application was sufficient. Cultures with a lot of spores tended to give slightly more species identifications in BDAL regardless of the age of cultures. Filamentous Fungi Library, in some cases, tended to improve the performance compared to BDAL for younger cultures. More studies are required to evaluate and optimize MALDI-TOF MS as a method of mold identification.

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  • 3.
    Oliveira Ivarsson, Martin
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Verifiering av P-LDL-kolesterol på Beckman Coulter AU6802019Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Cholesterol is transported in the blood by lipoproteins. High levels of low-density lipoprotein (LDL)-cholesterol in the blood is a risk factor for cardiovascular disease. The concentration of LDL-cholesterol can be calculated using the Friedewald formula but there are also methods that measure LDL-cholesterol directly. The aim of this study was to verify the method P-LDL-cholesterol on a Beckman Coulter AU680 analyzer. Within-run imprecision and total imprecision were analyzed. The correlation between direct LDL-cholesterol and calculated LDL-cholesterol was examined using 43 patient samples with triglyceride levels < 4,5 mmol/L and 11 patient samples with triglyceride levels > 4,5 mmol/L. The Friedewald formula is not supposed to be used on triglyceride levels > 4,5 mmol/L, but in this case the formula was used anyway to evaluate differences between the methods at high triglyceride concentrations. The coefficient of variation (CV) for the within-run imprecision was about 0,5 %, both for the low control (A1) and the high control (A2). Total imprecision had a CV of 1,21 % for A1 and 1,11 % for A2. There was a linear relationship between the methods, but the direct method gave slightly higher results at low concentrations and slightly lower results at high concentrations compared to calculated LDL-cholesterol. At triglyceride levels > 4,5 mmol/L the results from the direct method was considerably higher than calculated LDL-cholesterol. The conclusion is that the precision of the method was good. The correlation between the results from direct LDL-cholesterol and calculated LDL-cholesterol was relatively high for samples with triglyceride levels < 4,5 mmol/L. At triglyceride levels > 4,5 mmol/L there was a big difference between the methods, probably because of falsely low results from calculated LDL-cholesterol.

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  • 4.
    Sonesson, Hannah
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Immunhistokemi - Utvärdering av antikropp mot pHH3 som potentiell markör för mitos vid diagnostisering av duktal bröstcancer2017Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Ductal carcinoma of the breast is the most common form of invasive breast tumours. The grading system for breast cancer is defined by Elston and Ellis and is based on three criterions. One of these criterions is the mitotic count in pathological sections of breast carcinomas stained with Hematoxylin Eosin. A common method often applied as a complement in diagnosis of breast carcinoma is immunohistochemical staining with use of antibodies directed against Ki67, a proliferation marker. Phosphohistone H3 is a histone protein that is located in the cell nucleus. The protein is believed to be a specific marker for mitosis since it only is phosphorylated during mitosis, and to some extent at the end of the G2-phase. The purpose of this study was to evaluate pHH3 as a potential marker for mitosis when diagnosing ductal breast cancer. The purpose was also to compare the method to mitotic figuring and the count of Ki67-positive cells, and to study the inter-individual variability when assessing the histological sections. The material consisted of 20 biopsies containing invasive ductal breast cancer. The sections were stained using IHC and all sections were evaluated microscopically. Cells positive for pHH3, Ki67 and mitotic cells were quantified, by three doctors. From the doctors results an average value was determined for each case and method. To be able to compare the methods the coefficient of variation was calculated. The average value of the coefficient of variation was determined for each method and also the standard deviation (SD). The coefficient of variation showed average values of 0,21 for Ki67 +/- 0,10 SD, 0,33 for pHH3 +/- 0,14 SD and 0,46 for mitotic figuring +/- 0,34 SD. The correlation coefficients for the methods and each doctor showed dispersion. The correlations showed average values of r = 0,78 for Ki67 and pHH3, r = 0,74 for Ki67 and mitosis and r = 0,83 for pHH3 and mitosis. According to this study it seems as though anti-pHH3 could complement the other methods. However explicit criteria which defines a threshold value of which cells should be considered pHH3-positive needs to be established. The inter-individual differences seem to decrease using antipHH3 compared with mitotic counting, which is more time consuming. Although the minimum difference can be seen when assessing anti-Ki67 as a proliferation marker.

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  • 5.
    Tornegård, Mimmie
    Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
    Utvärdering av ett nyutvecklat ELISA-kit för analys av fritt kalcidiol: samt dess korrelation med totalt kalcidiol i serum (LC-MS/MS)2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Studies have recently shown that vitamin D deficiency is a more common condition than previously assumed. It is the protein-bound pro-hormone calcidiol in serum that is analysed when the vitamin D status assessed. The largest fraction of calcidiol is bound to plasma proteins, but a very small fraction calcidiol circulate free in plasma. The last decade’s researches have had an interest in the free fraction which can possibly be more biologically relevant for the assessment of vitamin D status. An Enzyme-Linked Immunosorbent Assay (ELISA) method is introduced that can measure the amount of free calcidiol in serum. The purpose of this study was to compare the new ELISA method with a previous studies result, where in protein bound calcidiol was measured by LC-MS/MS. Study participants were 46 individuals, 20 men and 26 women. The average and standard deviation for total calcidiol was 74.2 nmol/L and 18.8 nmol/L and for free calcidiol it was 5.0 pg/mL and1.4 pg/mL. The results showed that the method had a total imprecision (CV%) between two samples of 6.0 %. The total imprecisionen for the high control was 26.6 % and for the low control it was 17.8 %. The coefficient of determination between the LC-MS/MS and ELISA was weak, R2 = 0.114. The weak correlation might have been caused by the small spread of calcidiol concentrations in the group studied, as well as the limited number of individuals sampled. In a manual ELISA are drift and pipetting errors not unusual, because the analysis includes several manual steps and can be minimized with automation of the method. Studies of various disease groups, with varying vitamin D status and more analyses of free and protein bound calcidiol are needed to establish the relevance of measuring free calcidiol as a marker of vitamin D status.

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