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  • 1. Abdelfatah Possnert, Heba
    Detection of Thymidine Kinase 1 Activity in Whole Blood Using an Oligonucleotide System2014Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    In today’s medical science studies, many tumor markers are being used to monitor cancer cell proliferation, but the number of assays for analysis of these markers are few. The aim of this study was to find an easier and more time-efficient way to measure the activity of a specific tumor marker called tymidine kinase 1 (TK1). This tumor marker is an important enzyme involved in cell proliferation and is a key enzyme in the salvage pathway. TK1 activity is related to the occurrence of hematological malignancies and cell activity and therefore have been used as a marker when monitoring this group of patients in treatment. Measurement of the enzyme activity in this study was performed by using an oligonucleotide assay. Detection of the enzyme activity in whole blood and in plasma has not previously been shown. The TK1 activity measured in whole blood and plasma correlated with TK1 activity measured in serum (R2=0,8651 and R2 =0,9845, respectively). It was found that it is possible to determine the TK1 activity in whole blood but only if the activity was measured on the same day as the blood samples were taken. The results shows that the activity measurement of TK1 in plasma and whole blood can be used as a marker to verify patients' therapy in cancer care. This study is only the beginning and further investigations should be made in the future to determine if the method that is subject to this study has the requested effects.

  • 2.
    Abelson, Klas
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience.
    Acetylcholine in Spinal Pain Modulation: An in vivo Study in the Rat2005Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The spinal cord is an important component in the processing and modulation of painful stimuli. Nerve signals from the periphery are relayed and further conducted to the brain (nociception) in the spinal cord, and the most essential modulation of painful information (antinociception) occurs here. Several neurotransmitters are involved in spinal pain modulation, among them acetylcholine. However, the role of acetylcholine has previously been little investigated.

    In the present thesis, the acetylcholine release in the spinal cord was studied in vivo. By using spinal microdialysis on anaesthetised rats, the effects on the intraspinal acetylcholine release of various receptor ligands and analgesic agents were examined. This, together with pain behavioural tests and in vitro pharmacological assays, was used to evaluate the role of acetylcholine in spinal pain modulation. The four studies in this thesis resulted in the following conclusions:

    An increased release of spinal acetylcholine is associated with an elevated pain threshold, while a decreased acetylcholine release is associated with hyperalgesia, as seen after systemic treatment with a muscarinic agonist and an antagonist.

    Lidocaine is a potent analgesic when given systemically. It was found to produce an increase of intraspinal acetylcholine after intravenous injection of analgesic doses. This effect was attenuated after muscarinic, and abolished after nicotinic, receptor blockade.

    Various a2-adrenergic ligands, associated with nociceptive or antinociceptive effects, were found to affect intraspinal acetylcholine release via action on nicotinic receptors.

    Finally, the involvement of spinal acetylcholine in the analgesic effects of aspirin and paracetamol was examined. It was found that spinal acetylcholine could participate in the analgesic effects of aspirin, but not of paracetamol.

    The present thesis provides data that clearly demonstrate a relationship between intraspinal acetylcholine and antinociception, and elucidate interactions between acetylcholine and other mechanisms that mediate antinociception in the spinal cord.

    List of papers
    1. Intravenously administered oxotremorine and atropine, in doses known to affect pain threshold, affect the intraspinal release of acetylcholine in rats
    Open this publication in new window or tab >>Intravenously administered oxotremorine and atropine, in doses known to affect pain threshold, affect the intraspinal release of acetylcholine in rats
    2002 (English)In: Pharmacology and Toxicology, ISSN 0901-9928, E-ISSN 1600-0773, Vol. 90, no 4, p. 187-192Article in journal (Refereed) Published
    Abstract [en]

    Abstract:Both systemically and intrathecally administered cholinergic agonists produce antinociception while cholinergic antagonists decrease pain threshold. The mechanism and the site of action of these substances are not known. In the present study it was hypothesized that systemically administered muscarinic agonists and antagonists modify nociceptive threshold by affecting intraspinal release of acetylcholine (ACh). Catheters were inserted into the femoral vein in rats maintained on isoflurane anaesthesia for administration of oxotremorine (10–300 μg/kg) and atropine (0.1, 10, 5000 μg/kg). Spinal microdialysis probes were placed intraspinally at approximately the C2–C5 spinal level for sampling of acetylcholine and dialysis delivery of atropine (0.1, 1, 10 nM). Additionally, the tail-flick behaviour was tested on conscious rats injected intraperitoneally with saline, atropine (10, 100 and 5000 μg/kg), or subcutaneously with oxotremorine (30, 100, 300 μg/kg). Subcutaneous administration of oxotremorine (30, 100, 300 μg/kg) significantly increased the tail-flick latency. These doses of oxotremorine dose-dependently increased the intraspinal release of acetylcholine. Intravenously administered atropine, in a dose that produced hyperalgesia (5000 μg/kg) in the tail-flick test, significantly decreased the intraspinal release of acetylcholine. Our results suggest an association between pain threshold and acetylcholine release in spinal cord. It is also suggested that an approximately 30% increase in basal ACh release produces antinociception and that a 30% decrease in basal release produces hyperalgesia.

    Place, publisher, year, edition, pages
    Blackwell, 2002
    National Category
    Biomedical Laboratory Science/Technology
    Identifiers
    urn:nbn:se:uu:diva-7215 (URN)10.1034/j.1600-0773.2002.900403.x (DOI)
    Available from: 2006-10-18 Created: 2006-10-18 Last updated: 2017-12-14Bibliographically approved
    2. Intravenously administered lidocaine in therapeutic doses increases the intraspinal release of acetylcholine in rats
    Open this publication in new window or tab >>Intravenously administered lidocaine in therapeutic doses increases the intraspinal release of acetylcholine in rats
    2002 (English)In: Neuroscience Letters, ISSN 0304-3940, E-ISSN 1872-7972, Vol. 317, no 2, p. 93-6Article in journal (Refereed) Published
    Abstract [en]

    The local anesthetic lidocaine suppresses different pain conditions when administered systemically. Part of the antinociceptive effect appears to be mediated via receptor mechanisms. We have previously shown that muscarinic and nicotinic agonists that produce antinociception increase the intraspinal release of acetylcholine. In the present study it was hypothesized that systemically administered lidocaine is acting through the same mechanisms as cholinergic agonists and affects the intraspinal release of acetylcholine. Microdialysis probes were placed in anesthetized rats for sampling of acetylcholine. Ten and 30 mg/kg lidocaine injected intravenously significantly increased the intraspinal release of acetylcholine. The effect of lidocaine could be reduced by pretreatment with intraspinally administered atropine or mecamylamine. Our results suggest that the antinociceptive effect produced by systemically administered lidocaine is mediated through an action on muscarinic and nicotinic receptors.

    Place, publisher, year, edition, pages
    Elsevier, 2002
    Keywords
    Lidocaine; Spinal cord; Pain; Microdialysis; Acetylcholine; Muscarinic; Nicotinic
    National Category
    Biomedical Laboratory Science/Technology
    Identifiers
    urn:nbn:se:uu:diva-7227 (URN)10.1016/S0304-3940(01)02440-5 (DOI)
    Available from: 2006-10-24 Created: 2006-10-24 Last updated: 2017-12-14Bibliographically approved
    3. The effects of the alpha2-adrenergic receptor agonists clonidine and rilmenidine, and antagonists yohimbine and efaroxan, on the spinal cholinergic receptor system in the rat
    Open this publication in new window or tab >>The effects of the alpha2-adrenergic receptor agonists clonidine and rilmenidine, and antagonists yohimbine and efaroxan, on the spinal cholinergic receptor system in the rat
    2004 (English)In: Basic & Clinical Pharmacology & Toxicology, ISSN 1742-7835, E-ISSN 1742-7843, Vol. 94, no 4, p. 153-60Article in journal (Refereed) Published
    Abstract [en]

    Cholinergic agonists produce spinal antinociception via mechanisms involving an increased release of intraspinalacetylcholine. The cholinergic receptor system interacts with several other receptor types, such as a2-adrenergic receptors.To fully understand these interactions, the effects of various receptor ligands on the cholinergic system must be investigatedin detail. This study was initiated to investigate the effects of the a2-adrenergic receptor agonists clonidine and rilmenidineand the a2-adrenergic receptor antagonists yohimbine and efaroxan on spinal cholinergic receptors in the rat. Spinalmicrodialysis was used to measure in vivo changes of acetylcholine after administration of the ligands, with or withoutnicotinic receptor blockade. In addition, in vitro binding properties of the ligands on muscarinic and nicotinic receptorswere investigated. It was found that clonidine and rilmenidine increased, while yohimbine decreased spinal acetylcholinerelease. Efaroxan affected acetylcholine release differently depending on concentration. Nicotinic receptor blockade atten-uated the effect of all ligands. All ligands showed poor binding affinity for muscarinic receptors. On the other hand, allligands possessed affinity for nicotinic receptors. Clonidine and yohimbine binding was best fit to a one site binding curveand rilmenidine and efaroxan to a two site binding curve. The present study demonstrates that the tested a2-adrenergicreceptor ligands affect intraspinal acetylcholine release in the rat evoked by nicotinic receptor mechanisms in vivo, andthat they possess binding affinity to nicotinic receptors in vitro. The binding of a2-adrenergic receptor ligands to nicotinicreceptors might affect the intraspinal release of acetylcholine.

    Place, publisher, year, edition, pages
    Blackwell, 2004
    National Category
    Biomedical Laboratory Science/Technology
    Identifiers
    urn:nbn:se:uu:diva-92739 (URN)10.1111/j.1742-7843.2004.pto940401.x (DOI)15078339 (PubMedID)
    Available from: 2005-03-30 Created: 2005-03-30 Last updated: 2017-12-14Bibliographically approved
    4. Spinal cholinergic involvement after treatment with aspirin and paracetamol in rats
    Open this publication in new window or tab >>Spinal cholinergic involvement after treatment with aspirin and paracetamol in rats
    2004 In: Neurosci. Lett., ISSN 0304-3940, Vol. 368, no 1, p. 116-120Article in journal (Refereed) Published
    Identifiers
    urn:nbn:se:uu:diva-92740 (URN)
    Available from: 2005-03-30 Created: 2005-03-30Bibliographically approved
  • 3.
    Abelson, Klas
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Hau, Jann
    Carlsson, Hans-Erik
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Undergraduate and postgraduate students' responses to mandatory courses (FELASA category C) in laboratory animal science 1997-20032005In: Internationalisation and Harmonisation of Laboratory Animal Care and Use Issues: Proceedings of the Ninth FELASA Symposium 14-17 June 2004, Nantes, France / [ed] M. R. Gamble, 2005Conference paper (Other academic)
  • 4.
    Abelson, Klas
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Höglund, Urban
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Intravenously administered lidocaine in therapeutic doses increases the intraspinal release of acetylcholine in rats2002In: Neuroscience Letters, ISSN 0304-3940, E-ISSN 1872-7972, Vol. 317, no 2, p. 93-6Article in journal (Refereed)
    Abstract [en]

    The local anesthetic lidocaine suppresses different pain conditions when administered systemically. Part of the antinociceptive effect appears to be mediated via receptor mechanisms. We have previously shown that muscarinic and nicotinic agonists that produce antinociception increase the intraspinal release of acetylcholine. In the present study it was hypothesized that systemically administered lidocaine is acting through the same mechanisms as cholinergic agonists and affects the intraspinal release of acetylcholine. Microdialysis probes were placed in anesthetized rats for sampling of acetylcholine. Ten and 30 mg/kg lidocaine injected intravenously significantly increased the intraspinal release of acetylcholine. The effect of lidocaine could be reduced by pretreatment with intraspinally administered atropine or mecamylamine. Our results suggest that the antinociceptive effect produced by systemically administered lidocaine is mediated through an action on muscarinic and nicotinic receptors.

  • 5.
    Abelson, Klas
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Höglund, Urban
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Intravenously administered oxotremorine and atropine, in doses known to affect pain threshold, affect the intraspinal release of acetylcholine in rats2002In: Pharmacology and Toxicology, ISSN 0901-9928, E-ISSN 1600-0773, Vol. 90, no 4, p. 187-192Article in journal (Refereed)
    Abstract [en]

    Abstract:Both systemically and intrathecally administered cholinergic agonists produce antinociception while cholinergic antagonists decrease pain threshold. The mechanism and the site of action of these substances are not known. In the present study it was hypothesized that systemically administered muscarinic agonists and antagonists modify nociceptive threshold by affecting intraspinal release of acetylcholine (ACh). Catheters were inserted into the femoral vein in rats maintained on isoflurane anaesthesia for administration of oxotremorine (10–300 μg/kg) and atropine (0.1, 10, 5000 μg/kg). Spinal microdialysis probes were placed intraspinally at approximately the C2–C5 spinal level for sampling of acetylcholine and dialysis delivery of atropine (0.1, 1, 10 nM). Additionally, the tail-flick behaviour was tested on conscious rats injected intraperitoneally with saline, atropine (10, 100 and 5000 μg/kg), or subcutaneously with oxotremorine (30, 100, 300 μg/kg). Subcutaneous administration of oxotremorine (30, 100, 300 μg/kg) significantly increased the tail-flick latency. These doses of oxotremorine dose-dependently increased the intraspinal release of acetylcholine. Intravenously administered atropine, in a dose that produced hyperalgesia (5000 μg/kg) in the tail-flick test, significantly decreased the intraspinal release of acetylcholine. Our results suggest an association between pain threshold and acetylcholine release in spinal cord. It is also suggested that an approximately 30% increase in basal ACh release produces antinociception and that a 30% decrease in basal release produces hyperalgesia.

  • 6.
    Abelson, Klas
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Höglund, Urban
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    The effects of the alpha2-adrenergic receptor agonists clonidine and rilmenidine, and antagonists yohimbine and efaroxan, on the spinal cholinergic receptor system in the rat2004In: Basic & Clinical Pharmacology & Toxicology, ISSN 1742-7835, E-ISSN 1742-7843, Vol. 94, no 4, p. 153-60Article in journal (Refereed)
    Abstract [en]

    Cholinergic agonists produce spinal antinociception via mechanisms involving an increased release of intraspinalacetylcholine. The cholinergic receptor system interacts with several other receptor types, such as a2-adrenergic receptors.To fully understand these interactions, the effects of various receptor ligands on the cholinergic system must be investigatedin detail. This study was initiated to investigate the effects of the a2-adrenergic receptor agonists clonidine and rilmenidineand the a2-adrenergic receptor antagonists yohimbine and efaroxan on spinal cholinergic receptors in the rat. Spinalmicrodialysis was used to measure in vivo changes of acetylcholine after administration of the ligands, with or withoutnicotinic receptor blockade. In addition, in vitro binding properties of the ligands on muscarinic and nicotinic receptorswere investigated. It was found that clonidine and rilmenidine increased, while yohimbine decreased spinal acetylcholinerelease. Efaroxan affected acetylcholine release differently depending on concentration. Nicotinic receptor blockade atten-uated the effect of all ligands. All ligands showed poor binding affinity for muscarinic receptors. On the other hand, allligands possessed affinity for nicotinic receptors. Clonidine and yohimbine binding was best fit to a one site binding curveand rilmenidine and efaroxan to a two site binding curve. The present study demonstrates that the tested a2-adrenergicreceptor ligands affect intraspinal acetylcholine release in the rat evoked by nicotinic receptor mechanisms in vivo, andthat they possess binding affinity to nicotinic receptors in vitro. The binding of a2-adrenergic receptor ligands to nicotinicreceptors might affect the intraspinal release of acetylcholine.

  • 7.
    Abelson, Klas
    et al.
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Neuroscience, Comparative Medicine.
    Kommalage, Mahinda
    Höglund, Urban
    Spinal cholinergic involvement after treatment with aspirin and paracetamol in rats2004In: Neuroscience Letters, ISSN 0304-3940, Vol. 368, no 1, p. 116-120Article in journal (Refereed)
  • 8.
    Adelholt, Denise
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    The Effects of Cell Culture Medium and Supplements on the Differentiation of Boundary Cap Neural Crest Stem Cells2016Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Boundary cap neural crest stem cells (bNCSCs) are multipotent cells that form a barrier between CNS and PNS, playing an important role in ingrowth of neurites into the spinal cord during development. Because of the stemness and multipotency of bNCSCs, they self-renew and can be used directly for transplantation or as a source of matured neural cells. It is important that cells used for cell therapy differentiate and develop into the mature cells that the recipient needs. To ensure this, cells are guided towards specific cell fates, and one way of doing this is with medium supplements. The purpose of this study was to analyze the effects of three different media with supplements on the differentiation of bNCSCs. Two cell lineages of bNCSCs expressing green- and red fluorescent protein were treated with different media for differentiation. The effects of the media supplements neurotrophic glial cell line-derived neurotrophic factor (GDNF), cilinary neurotrophic factor (CTNF) and fetal bovine serum (FBS) were compared, with one medium containing no additional factors. It was found that when GDNF and CTNF are supplemented in the differentiation media, bNCSCs are guided towards astrocytes. Interestingly, the medium containing no additional factors gave rise to an even amount of neurons and astrocytes. FBS had an inhibitory effect on overall differentiation of bNCSCs, giving rise to the smallest amount of neurons and astrocytes. The bNCSCs are promising for cell therapy, as their differentiation can be guided with the use of medium supplements.

  • 9. Adinda, Mathia
    Nutrition supplements when undergoing orthopedic surgery2014Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Malnutrition is prevalent in elderly populations with orthopedic disabilities, which is especially critical during surgery when the body is under much stress. It is important that these patients are well nourished to be able to cope when mechanisms such as immune system are activated. Nutrition is also important after surgery when the body is healing and sometimes struggling against different complications as infections. The aim of this study was to evaluate if nutrition supplements decreases the time needed for rehabilitation and improve the outcome after orthopedic surgeries. This was performed by analyzing biomarkers involved in wound healing. The study population comprised of 100 surgical patients at the age of 50 or older. The participant where divided into two groups, one test group that received nutrition supplements and one control group who did not receive any extra nutrition. Sandwich ELISA was used to measure myostatin, cathepsin S and cathepsin B concentrations in patient serum before and after surgery. There were no significant difference between the control group and the test group for any of the three biomarkers. The conclusion is that nutrition supplement does not decrease the rehabilitation time and outcome according to the results in this study.

  • 10. Adlerz, Linda
    Immunohistochemistry in an automated platform forPrion Protein in Transmissible Spongiform Encephalopathy2019Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Background: Transmissible spongiform encephalopathies, like Creutzfeldt-Jakobs Diseaseare fatal neurodegenerative disorders. They are caused by misfolding and aggregation of anendogenously expressed protein, prion. Detection of the pathological form of prions byimmunohistochemistry has become a valuable tool for diagnostics. However,immunohistochemistry of prions is traditionally performed manually and with various tediouspretreatments.Purpose: In clinical diagnostics fast, specific, reliable and standardised protocols are needed.In this study an optimised protocol for prion immunohistochemistry in an automated platformwas developed.Material and methods: Sections of paraffin-embedded brain tissue from patients with andwithout sporadic Creutzfeldt-Jakobs Disease was used for comparison withimmunohistochemistry with two different anti-prion antibodies 12F10 and 3F4. The effect ofdifferent pre-treatments like vaporised cooking under pressure, use of proteinase K, citratebuffer, guanidine thiocyanate and picric acid was investigated.Results: Both antibodies displayed similar patterns of immunoreactivity in the manual as wellas in the automated platform. Vaporised cooking under pressure was preferred before heatingin the automated platform due to minimal change in tissue morphology with the former. Theuse of citrate buffer and proteinase K was essential for detecting immunoreactivity whereasguanidine thiocyanate and picric acid could be excluded.Conclusion: Both 12F10 and 3F4 can be used for immunohistochemistry on an automatedplatform for clinical diagnostics. Pre-treatments in this protocol include formic acid,vaporised cooking under pressure in citrate buffer and the use of proteinase K.

  • 11.
    Ahlgren, Evelina
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Retrospective serological and virological survey of influenza D virus among cattle in Sweden2019Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Respiratory diseases in cattle can cause economic losses due to the decreased dairy and meat production. Virus is the main reason for these diseases. Symptoms can be fever, cough and nasal discharge.

        Influenza are a group of viruses belonged in the Ortomyxoviridae family. The big influenza groups are influenza A, B and C. The viruses can cause respiratory signs, and mammals can be affected. Recently a new influenza virus was found in the United States. The influenza virus was found in swine, but the natural host was later considered to be cattle. The virus was named influenza D. Different studies worldwide have confirmed the virus in a variety of regions. Antibodies have also been reported.

        In this study, virologic and serologic methods were used to detect if influenza D circulates among cattle in Sweden. The serologic method performed was indirect ELISA. Serum and milk samples were investigated in the ELISA method. For the virologic detection a real-time RT-PCR was made, with a variety of study material.

        Antibodies against influenza D were found in both serum and milk samples. No virus was found in the real-time RT-PCR. In Sweden the animal keeping is different compared to several other nations. For instance, the conditions of health and hygiene are better in Sweden, this may be an important cause of a system more resistant against spreading of infections. Influenza D could be more common in Sweden, but in that case further researches are needed to determine the prevalence.

  • 12.
    Ajalloueian, Fatemeh
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Polymer Chemistry. Uppsala University, Science for Life Laboratory, SciLifeLab.
    Zeiai, Said
    Fossum, Magdalena
    Hilborn, Jöns G.
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Polymer Chemistry. Uppsala University, Science for Life Laboratory, SciLifeLab.
    Constructs of electrospun PLGA, compressed collagen and minced urothelium for minimally manipulated autologous bladder tissue expansion2014In: Biomaterials, ISSN 0142-9612, E-ISSN 1878-5905, Vol. 35, no 22, p. 5741-5748Article in journal (Refereed)
    Abstract [en]

    Bladder regeneration based on minced bladder mucosa in vivo expansion is an alternative to in vitro culturing of urothelial cells. Here, we present the design of a hybrid, electrospun poly(lactic-co-glycolide) (PLGA) - plastically compressed (PC) collagen scaffold that could allow in vivo bladder mucosa expansion. Optimisation of electrospinning was performed in order to obtain increased pore sizes and porosity to consolidate the construct and to support neovascularisation and tissue ingrowth. Tensile tests showed an increase in average tensile strength from 0.6 MPa for PC collagen to 3.57 MPa for the hybrid construct. The optimised PLGA support scaffold was placed between two collagen gels, and the minced tissue was distributed either on top or both on top and inside the construct prior to PC; this was then cultured for up to four weeks. Morphology, histology and SEM demonstrated that the construct maintained its integrity throughout cell culture. Cells from minced tissue migrated, expanded and re-organised to a confluent cell layer on the top of the construct after two weeks and formed a multilayered urothelium after four weeks. Cell morphology and phenotype was typical for urothelial mucosa during tissue culture. (C) 2014 Elsevier Ltd. All rights reserved.

  • 13.
    Akgun, Kocere Kurdé
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Evaluation of Different Extraction- and Analysis Methods for Calprotectin in Feces2012Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Background Calprotectin is a protein expressed in the cytoplasm inside the neutrophile granulocytes. During inflammatory bowel disease (IBD), the neutrophile granulocytes are involved in a complex interaction at the inflammatory area where they die and release their content into the intestinal lumen. Therefore, calprotectin in stool is a suitable marker for diagnosis and measurement of the disease-activity in patients with IBD. The most commonly used method to detect calprotectin in stool is ELISA, but the process of manual preparation of stool samples is time-consuming.

    Aim The objective of the study was to evaluate an extraction method that could replace manual preparation of fecal samples and to compare different methods for measuring Calprotectin in stool using two ELISA-methods from two manufacturers and one rapidtest.

    Methods For extraction of calprotectin from stool samples we used sample collector tubes from Epitope Diagnostics and fecal preparation kits from Roche. Two different ELISA-kits for measuring calprotectin concentration in stool were compared. Measurements of calprotectin with rapid-test from Epitope Diagnostics were also performed and were compared with the two ELISA kits.

    Results The results indicate a poor correlation between two extraction methods with Sample Collector Tube and Roche preparation kit. The comparison between the two ELISA-kits showed poor correlation. Evaluation of rapid test showed 33% false negative results with a cut-off value at 50 mg/kg.

    Conclusion Evaluation of products from Epitope Diagnostics showed poor correlation with the Bühlmann ELISA and an unreliable rapid test. Therefore, none of evaluated products from Epitope Diagnostics is accurate enough to be used for clinical diagnosis in the laboratory.

  • 14.
    Akula, Srinivas
    et al.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
    Mohammadamin, Sayran
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology.
    Hellman, Lars
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Chemical Biology.
    Fc Receptors for Immunoglobulins and Their Appearance during Vertebrate Evolution2014In: PLoS ONE, ISSN 1932-6203, E-ISSN 1932-6203, Vol. 9, no 5, p. e96903-Article in journal (Refereed)
    Abstract [en]

    Receptors interacting with the constant domain of immunoglobulins (Igs) have a number of important functions in vertebrates. They facilitate phagocytosis by opsonization, are key components in antibody-dependent cellular cytotoxicity as well as activating cells to release granules. In mammals, four major types of classical Fc receptors (FcRs) for IgG have been identified, one high-affinity receptor for IgE, one for both IgM and IgA, one for IgM and one for IgA. All of these receptors are related in structure and all of them, except the IgA receptor, are found in primates on chromosome 1, indicating that they originate from a common ancestor by successive gene duplications. The number of Ig isotypes has increased gradually during vertebrate evolution and this increase has likely been accompanied by a similar increase in isotype-specific receptors. To test this hypothesis we have performed a detailed bioinformatics analysis of a panel of vertebrate genomes. The first components to appear are the poly-Ig receptors (PIGRs), receptors similar to the classic FcRs in mammals, so called FcRL receptors, and the FcR gamma chain. These molecules are not found in cartilagous fish and may first appear within bony fishes, indicating a major step in Fc receptor evolution at the appearance of bony fish. In contrast, the receptor for IgA is only found in placental mammals, indicating a relatively late appearance. The IgM and IgA/M receptors are first observed in the monotremes, exemplified by the platypus, indicating an appearance during early mammalian evolution. Clearly identifiable classical receptors for IgG and IgE are found only in marsupials and placental mammals, but closely related receptors are found in the platypus, indicating a second major step in Fc receptor evolution during early mammalian evolution, involving the appearance of classical IgG and IgE receptors from FcRL molecules and IgM and IgA/M receptors from PIGR.

  • 15.
    Albeer, Merna
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of ELISA and rapid test for the analysis of fecal Calprotectin2013Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    ABSTRACT

    Background Calprotectin is a protein found in the cytoplasm of neutrophile granulocytes. In the course of inflammatory bowel disease (IBD), calprotectin is released during chronic inflammation in the gut. Activation of neutrophils during the inflammation is followed by activation and secretion of pro-inflammatory molecules such as calprotectin. Calprotectin is stable in stool up to 7 days and can therefore be used as a non-invasive marker for diagnosis, treatment and measurement of the disease activity in patients with IBD. The most common method for analysis of calprotectin concentration is ELISA. This method is time-consuming and many manufactures have therefore developed rapid tests as a faster alternative for quantification of calprotectin in stool.

    Aim The aim of the study was to evaluate one ELISA and one rapid test from the same manufacture compare the data with the existing ELISA-method used in the laboratory for routine analysis.

    Methods A rapid test (CalFast) and an ELISA method (CalPrest) from Eurospital, were used for analysis of calprotectin in stool. These two methods were compared with known concentrations of calprotectin obtained by the ELISA method from Bühlmann used in the routine work. 

    Results The results showed poor correlation between the rapid test and the ELISA method. Furthermore, the comparison between the two ELISA-methods showed a poor correlation.

    Conclusion Evaluation of the two new methods showed poor correlation with the existing ELISA method from Bühlmann. Evaluation of the rapid test did not show any correlation with the two ELISA methods and the data cannot be trusted. It is difficult to conclude which of the two ELISA methods gives accurate results due to the absence of an international standard.

  • 16.
    Ali, Iman
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Pan Genera Detection-​test, a new bacterial detection systems for durability extension of platelets2013Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Platelets are blood cells that important for transfusion and blood coagulation. Patients will get transfusion with manufactured platelets if they have platelet disorder or extensive blood loss. Platelets isolated from 5 buffy-coats by different ways at a blood center are usable for about 5 days. The sustainability of platelets could be extended by various methods. The previously used method in Gävle BLC was the bacterial culture method using aerobic and anaerobic bottles, which took 5 days to receive an answer. Therefore, this project used Pan Genera Detection (PGD) test as an alternative method to extend the sustainability of platelets. The PGD ​​Test is a rapid method that only takes about 30 minutes. This study showed that PGD the test yielded good results to reduced costs compared with the old method. The durability was extended by 36 h and the analysis runs in routine now at Gävle blood center.

    Finally, this project has shown that PGD-test is a cheaper, faster and safer approach for patient health compared to the old method, the bacterial culture method.

  • 17. Allabwani, Haifaa
    Verification of Bordetella pertussis and Bordetella holmesii withreal-time PCR2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Bordetella pertussis causes whooping cough, which is a very dangerous disease especially for children. A correct diagnose is very important. Bordetella pertussis usually misidentifies with Bordetella holmesii by routine polymerase chain reaction.The aim of the study is to evaluate a target sequence that can verify a Bordetella pertussis infection and distinguish it from other Bordetella species, in specific Bordetella holmesii.The target sequences' sensitivities were tested on the control bacterial strains. Sixty-five clinical samples were analyzed with real-time PCR.The study concludes that both IS481 and ptxA target sequences are highly specific to detect B. pertussis, while both ho_IS1001 and IS481 are specific to detect B. holmesii. Pa_IS1001 is specific to detect B. parapertussis. Culturing is less sensitive to detect Bordetella infection than real-time PCR.

  • 18. Almeflo, Sandra
    The effect of shifting host plants on growth of butterfly larvae of Polygonia c-album.2014Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
  • 19. Alsaadi, Hani
    A new High Sensitive Functional Nephelometrical Assay for Assaying C- reactive protein in Serum Based on Phosphocholine Interaction2015Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Abstract

    C-reactive protein (CRP) is able to bind phosphocholine in the presence of calcium ions. According to a previous functional property of CRP, we tried to develop an affordable and cheap high sensitive nephelometric CRP assay using soy oil.

    Serum samples were measured by Nephelometer BNII (Siemens), by mixing the serum with diluted soy oil emulsion (Intralipid

    ®

    20%) and Tris-calcium buffer (PH 7.5). The measurement took place after 12 min incubation time at 37°C by measuring the agglutination between CRP and phosphocholine. Results from our automated functional assay were compared with results obtained using an immunoturbidimetric CRP assay.

    Results showed a good correlation coefficient for method comparison between functional nephelometric CRP assay and immunoturbidimetric CRP assay, r = 0.895, significance level p <0.0001. The limit of detection for the functional nephelometric CRP assay was 0.1 mg/L. However, the within run % CV values for the functional assay were 6.1 % (20 mg/L), 4.7 % (50 mg/L) and 4.5 % (100 mg/L). The between-run % CV values were 17.6 % (20 mg/L), 18.8 % (50 mg/L), and 11.3 % (100 mg/L).

    The new functional nephelometric CRP assay enables high sensitive CRP measurement in serum in the range of 0.1 mg/L to 300 mg/L. The functional assay could be used for veterinary analysis due to the ability to measure CRP according to the functional properties, not the morphological properties which depend on specific antibodies.

  • 20.
    Andersson, Daniel
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of Correlation between mRNA and Protein Expression of Tripeptidyl-Peptidase II: Possible Future Use as a Biomarker for Cancer?2013Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Cancer remains one of the most common causes for death in the world today. Researchers are continuously trying to improve old, and develop new, methods in order to strife this global problem. Much research is being made trying to find new specific biomarkers that can be used to detect and diagnose cancer in an early stage.

    One candidate protein for possible future use as a biomarker is tripeptidyl-peptidase II (TPPII) which has previously been shown to be up-regulated in Burkitt´s lymphoma. This paper focuses on the expression of TPPII on an mRNA-level to see if there is any difference between expression in human leucocytes from patients with a leukemia diagnosis and a healthy volunteer, in order to evaluate if the expression of TPPII have any future use as a biomarker.

    Patient samples were analyzed using real time qPCR, to study the expression of mRNA, and Western blot, in order to correlate the mRNA findings with protein expression. Three different cell lines with different characteristics regarding expression and function of TPPII were also used to validate the methods used and for comparison with the patient samples analyzed.

    A difference in expression of mRNA were seen between the different patient samples, both individually and between larger groups of samples with the same diagnosis, indicating a large individual variation, thus making future use in a clinical setting difficult. However, seeing as only a few samples were analyzed in this study, more research must be done in order to draw any final conclusions.

  • 21.
    Andersson, Frida
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Dementia; common cause of suicide among elderly?2006Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Elderly committing suicide can be in a “preclinical phase” of dementia. Depressive symptoms may indicate a risk to develop a disease of dementia, for example Alzheimer’s Disease. Today almost 10% of the Swedish population older than 65 years suffer from a cognitive impairment diagnosed as dementia. Symptoms of dementia are associated with degenerative changes in the brain caused by a deposition of amyloid, leading among others things to a nerve cell death. A clinical diagnosis can be hard to set, and a definitive diagnose can only be set after a pathological examination, which only is possible after death. For this study we used Congo red staining of brains sections to find amyloid in autopsies from elderly people committing suicide. 35 cases (>60 year) were studied. Of the 35 cases 1/3 showed to be positive for amyloid deposition. This result in addition to other studies suggest that depressive symptoms is a “preclinical phase” of dementia, and therefore the suicide risk for this group must be consider to be elevated. However, more reliable prospective studies most be done to confirm this retrospective study.

  • 22.
    Andersson, Jim
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    LEFT VENTRICULAR EJECTION FRACTION: A RETROSPECTIVE STUDY COMPARING 2D ECHOCARDIOGRAPHY AND GATED SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY (SPECT) IN CLINICAL USE.2009Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Objectives

    The aim of this study was to compare left ventricular ejection fraction (LVEF) results derived from gated single photon emission computed tomography (SPECT) using Cedars-Sinai quantitative gated SPECT (QGS) processing software with results from 2D echocardiography, both obtained in routine clinical diagnostic use.

     

    Methods

    Data from previously performed tests were obtained from 73 patients who had undergone both 2D echocardiography and gated SPECT within a time span of 6 months and had not had significant events that could influence LVEF. LVEF from 2D echocardiography was reassessed to obtain discrete values and then the data was compared using Bland-Altman analysis.

     

    Results

    The correlation between the tests was shown to be good, but precision lacked. Bland-Altman analysis showed a bias of -0.8 percentage points when gated SPECT compared to mean values and 2 standard deviations (SD) ranged from -20.2 to 18.6.

     

    Conclusions

    LVEF values from the two methods can differ quite a bit and comparisons between them should be done with great caution.

  • 23. Andersson, Sara
    Effects of the different surface nanotopography of Poly(MAA-DAP) on the activation of the cascade systems in blood2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Background: Biomaterials are used for transplants and dialyses. A biomaterial that is not well adjusted to the body but is inserted into the body, may cause serious thrombosis and thromboembolic events as well as inflammation. PVC hoses used in dialysis today causes substantial complement activation. Therefore, poly (MAA-DAP), a polymer that has been developed in to four different nanotopographies, could be a possible biomaterial used in haemodialysis and catheters.Aim: The aim of these nanostructures is to see if it can be used as a substitute for PVC, and to see if there is a difference between the four different structures. Thus, these nanotopographies should have a low thrombogenic effect and a low complement activation.Material and methods: To analyse the compatibility with the cascade system, the intrinsic pathway and the alternative pathway, blood samples were analysed by use of ELISA. Plasma was collected from the 2-well slide chamber blood experiments were the whole blood was added to a slide chamber. The amount of platelets was, in addition to ELISA, also analysed using Sysmex XP-300 cellcounter.Results: The results showed that the MAA-DAP polymer surfaces were complement activating but not platelet activating. There was no significant difference in complement activation between the different MAA-DAP polymer surfaces. The more structured wires and fibrils were slightly more platelet activating but the difference was not significant enough.Conclusion: It can be concluded that the nanotopographies do not have significant impact on blood, it is the polymer material that effects the blood. Future studies could be performed on other type of material with different topographies and with blood.

  • 24.
    Andersson, Vidar
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of CellaVision DM1200 Vet and its ability to differentiate feline leukocytes compared to manual differential count and Advia 21202016Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Leukocyte differential count in peripheral blood smear has, ever since the method was developed more than 100 years ago, been one of the most frequently used diagnostics tool in the routine hematology laboratory. The manual differential count of leukocytes using a microscope is still the standard method in most small and medium sized laboratories. Even though the method does not require any expensive instruments it comes at a high cost due to it being labor intensive and time consuming. In recent years the rapid technical advancements has led to the development of automatic or semi-automatic methods in which the leukocytes are differentiated. In this study a method comparison was made between manual leukocyte differential counts, CellaVision DM1200 Vet and Advia 2120 when analyzing 106 fresh, feline blood samples. The general agreement between results was good, especially for the most common leukocytes, such as neutrophils and lymphocytes. Results for eosinophils and monocytes had moderate agreement. The confidence intervals were generally wider when CellaVision DM1200 Vet was compared with Advia 2120, than when CellaVision DM1200 Vet was compared to the manual differential count. Despite the fact that Advia 2120 and CellaVision DM1200 Vet are both faster and often show comparable results to the manual differential count, the light microscopy will remain the gold standard for difficult samples, where there is suspicion of inflammation (band neutrophils), intracellular microorganisms, reactive lymphocytes or if the sample contains a high degree of smudge cells or artifacts. 

  • 25. Andreassen, Jenny
    PRESERVATION OF DNA AND BACTERIA FOR INCORPORATION IN CULTURE COLLECTIONS2015Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    This study contains of two different parts, an investigation of long time stored prokaryotic DNA for incorporation in a DNA-bank, and lyophilization of two Flavobacterium strains with different lyoprotectants and validation of an alternative method for determination of concentration. For the DNA-project PCR and capillary gel electrophoresis was used for sequencing, and lyophilization, CFU-counts and Start of growth time was used to investigate the Flavobacteriums. 87.5% of the DNA samples with maintained quality were correctly identified. The lyophilization was successful, with varying results from the revivals. The validation could not be completed due to incoherent results between the two methods.An alternative validation method is suggested. The presence of viable but non-cultivable should also be considerated for continued studies.

  • 26.
    Ask, Alexandra
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    REPLICATING THE TUMOUR MICROENVIRONMENT:CHEMOSENSITIVITY TESTING IN FIBROBLAST COCULTURES2017Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
  • 27.
    Attevall, Janine
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Genetic investigation of rare microdeletions and microduplications with distinct clinical features2018Independent thesis Advanced level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Chromosomal microduplications and microdeletions represent the largest significant fraction of Copy Number Variants (CNV) and some can cause developmental delay and intellectual disability. The relatively common microdeletions and microduplications syndromes are well known but there are clinical cases with unique microduplications/-deletions that have not been studied sufficiently. The aim of this study was to verify positive SNP-microarray results during the genetic investigation of four patients with different unique microduplications or microduplications with metaphase-FISH. Since sodium heparin tubes contain the existing test material for these patients, a validation of RNA extraction was also made to verify if blood taken in these tubes can be used for further gene expression analysis.

    Blood samples and amniotic fluid were analyzed with SNP-microarray and verified with metaphase-FISH. RNA was extracted from blood taken in EDTA and sodium heparin tubes from five different individuals. Gene expression analysis with RT-qPCR were performed as a control for the RNA extraction using the genes ISPD and GUSB.

    FISH-analysis could detect the chromosomal rearrangements in all patients and investigation of the parents showed that these rearrangements were de novo. These results contribute to a better understanding of these unique aberrations and the patients´ phenotypes. There was no significant difference in RNA quality between sodium heparin and EDTA tubes. However RT-qPCR showed lower efficiency for both target gene (ISPD) and reference gene (GUSB) in RNA samples extracted from sodium heparin tubes. Blood samples in sodium heparin tubes should therefore not be used for RNA-analysis in further investigations.

  • 28.
    Aulin, Cecilia
    Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Materials Chemistry.
    Extracellular Matrix Based Materials for Tissue Engineering2010Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The extracellular matrix is (ECM) is a network of large, structural proteins and polysaccharides, important for cellular behavior, tissue development and maintenance. Present thesis describes work exploring ECM as scaffolds for tissue engineering by manipulating cells cultured in vitro or by influencing ECM expression in vivo. By culturing cells on polymer meshes under dynamic culture conditions, deposition of a complex ECM could be achieved, but with low yields. Since the major part of synthesized ECM diffused into the medium the rate limiting step of deposition was investigated. This quantitative analysis showed that the real rate limiting factor is the low proportion of new proteins which are deposited as functional ECM. It is suggested that cells are pre-embedded in for example collagen gels to increase the steric retention and hence functional deposition.

    The possibility to induce endogenous ECM formation and tissue regeneration by implantation of growth factors in a carrier material was investigated. Bone morphogenetic protein-2 (BMP-2) is a growth factor known to be involved in growth and differentiation of bone and cartilage tissue. The BMP-2 processing and secretion was examined in two cell systems representing endochondral (chondrocytes) and intramembranous (mesenchymal stem cells) bone formation. It was discovered that chondrocytes are more efficient in producing BMP-2 compared to MSC. The role of the antagonist noggin was also investigated and was found to affect the stability of BMP-2 and modulate its effect. Finally, an injectable gel of the ECM component hyaluronan has been evaluated as delivery vehicle in cartilage regeneration. The hyaluronan hydrogel system showed promising results as a versatile biomaterial for cartilage regeneration, could easily be placed intraarticulary and can be used for both cell based and cell free therapies.

    List of papers
    1. Extracellular matrix-polymer hybrid materials produced in a pulsed-flow bioreactor system
    Open this publication in new window or tab >>Extracellular matrix-polymer hybrid materials produced in a pulsed-flow bioreactor system
    2009 (English)In: Journal of Tissue Engineering and Regenerative Medicine, ISSN 1932-6254, Vol. 3, no 3, p. 188-195Article in journal (Refereed) Published
    Abstract [en]

    Cell adhesion, interaction with material, cell proliferation and the production of an extracellular matrix (ECM) are all important factors determining the successful performance of an engineered scaffold. Scaffold design should aim at creating structures which can guide cells into forming new, functional tissue. In this study, the concept of in situ deposition of ECM by human dermal fibroblasts onto a compliant, knitted poly (ethyleneterephtalate) support is demonstrated, creating in vitro produced ECM polymer hybrid materials for tissue engineering. Comparison of cells cultured under static and dynamic conditions were examined, and the structure and morphology of the materials so formed were evaluated, along with the amount collagen deposited by the seeded cells. In vitro produced ECM polymer hybrid scaffolds could be created in this way, with the dynamic culture conditions increasing ECM deposition. Histological analysis indicated a homogenous distribution of cells in the 1 mm thick scaffold, surrounded by a matrix-like structure. ECM deposition was observed throughout the materials wigh 81.6 µg/cm2 of collagen deposited after 6 weeks. Cell produced bundles of ECM fibres bridged the polymer filaments and anchored cells to the support. These findings open hereto unknown possibilities of producing materials with structure designed by engineering together with biochemical composition given by cells.

    Place, publisher, year, edition, pages
    John Wiley & Sons, Ltd, 2009
    Keywords
    extracellular matrix, scaffold, polymer support, fibroblasts, bioreactor, dynamic culture conditions
    National Category
    Chemical Sciences
    Research subject
    Inorganic Chemistry
    Identifiers
    urn:nbn:se:uu:diva-106096 (URN)10.1002/term.152 (DOI)000265268400003 ()
    Available from: 2009-06-15 Created: 2009-06-15 Last updated: 2010-08-02Bibliographically approved
    2. Bulk collagen incorporation rates into knitted stiff fibre polymer in tissue-engineered scaffolds: the rate-limiting step
    Open this publication in new window or tab >>Bulk collagen incorporation rates into knitted stiff fibre polymer in tissue-engineered scaffolds: the rate-limiting step
    Show others...
    2008 (English)In: Journal of Tissue Engineering and Regenerative Medicine, ISSN 1932-6254, Vol. 2, no 8, p. 507-514Article in journal (Refereed) Published
    Abstract [en]

    Fabrication of tissue-engineered constructs in vitro relies on sufficient synthesis of extracellular matrix (ECM) by cells to form a material suitable for normal function in vivo. Collagen synthesis by human dermal fibroblasts grown in vitro on two polymers, polyethylene terephthalate (PET) and polyglycolic acid (PGA), was measured by high-performance liquid chromatography (HPLC). Cells were either cultured in a dynamic environment, where meshes were loaded onto a pulsing tube in a bioreactor, or in a static environment without pulsing. Collagen synthesis by cells cultured on a static mesh increased by six-fold compared to monolayer culture, and increased by up to a further 5.4-fold in a pulsed bioreactor. However, little of the collagen synthesized was deposited onto the meshes, almost all being lost to the medium. The amount of collagen deposited onto meshes was highest when cells were cultured dynamically on PET meshes (17.6 µg), but deposition still represented only 1.4% of the total synthesized. Although total collagen synthesis was increased by the use of 3D culture and the introduction of pulsing, the results suggest that the limiting factor for fabrication of a tissue-engineered construct within practical timescales is not the amount of collagen synthesized but the quantity retained (i.e. deposited) within the construct during culture. This may be enhanced by systems which promote or assemble true 3D multi-layers of cells.

    Place, publisher, year, edition, pages
    John Wiley & Sons, Ltd., 2008
    Keywords
    collagen synthesis, collagen deposition, tissue engineering, polyethylene terephthalate, 3D culture, bioreactors
    National Category
    Chemical Sciences
    Research subject
    Inorganic Chemistry
    Identifiers
    urn:nbn:se:uu:diva-99347 (URN)10.1002/term.126 (DOI)000262272900007 ()
    Available from: 2009-03-12 Created: 2009-03-12 Last updated: 2010-08-04Bibliographically approved
    3. Comparative studies on BMP-2 processing and secretion in chondrocytes and mesenchymal cells and the effect of noggin
    Open this publication in new window or tab >>Comparative studies on BMP-2 processing and secretion in chondrocytes and mesenchymal cells and the effect of noggin
    (English)Manuscript (preprint) (Other academic)
    Keywords
    bone formation, BMP-2, noggin, chondrocytes, mesenchymal stem cells
    Identifiers
    urn:nbn:se:uu:diva-110746 (URN)
    Available from: 2009-11-24 Created: 2009-11-24
    4. Evaluation of an injectable hyaluronan hydrogel for cartilage regeneration
    Open this publication in new window or tab >>Evaluation of an injectable hyaluronan hydrogel for cartilage regeneration
    Show others...
    (English)Manuscript (preprint) (Other academic)
    Keywords
    hyaluronan, chondrocytes, mesenchymal stem cells, cartilage, injectable, bone morphogenetic protein-2, rabbit model
    Identifiers
    urn:nbn:se:uu:diva-110750 (URN)
    Available from: 2009-11-24 Created: 2009-11-24
  • 29. Aymara, Tagyzade
    Molecular expression of receptive stage endometriumin healthy women and women with endometriosis2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Endometriosis affects about 10-15% of women in their reproductive age, which increases the risk for infertility. The molecular expression profile of receptive endometrium of women with endometriosis may differ from that of receptive endometrium of healthy women. Thus, the objective of this study was to examine the molecular expression in endometrium of women with endometriosis and compare it with the endometrium from healthy women collected during receptive phase.Endometrium was collected during receptive phase from women with endometriosis (n=4) and from healthy women with proven fertility (n=8). Paraformaldehyde fixed and paraffin embedded tissues were sectioned and the differential protein expression of SOX17, ezrin, WT1 and SLPI were studied by use of immunohistochemistry, and analysed under light microscopy for staining intensity and area. Mann-Whitney U-test was performed to find differences in protein staining.We found that protein expression of SOX17 was confined to endometrial glands. The expression of ezrin present in glands was significantly higher in endometrium from women with endometriosis (p<0.01). There were no differences between the two groups in the expression of SOX17 or WT1. We could not find any detectable levels of SLP1 in the endometrial sections. Thus, we conclude that the endometrium of women with endometriosis have differential expression of ezrin.

  • 30. Aziz, Sheima
    Real-time PCR detection of MRSA on the CepheidGeneXpert®System2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Background: MRSA or "Meticillin-resistant Staphylococcus aureus" has become a healthcareissue around the world. Over the last few years, MRSA infections has increased in 520 casesand become more common in society, causing a life threating infection and healthcareproblem. In order to avoid spreading of MRSA between patients in hospitals, specific analysisare necessary to monitor the bacteria.Aim: The purpose of the project was to validate NxG-kit in GeneXpert-instruments toverify and compare this kit to the Nasal Complete kit and to SYBR-Green based on in-housereal-time PCR. This comparison was made to see if NxG-kits can detect the SCCmec carryingMRSA's mecA and mecC-gene to replace in-house PCR and Nasal Complete-kits with NxGkits.Material and methods: In this project, some analyses such as in-house real time PCR andGeneXpert systems were performed using patient’s samples from different locations includingsome strains to detect specific genes in MRSA at the microbiology laboratory (Unilabs) inStockholm.Result: The result of the analysis performed with NxG-kit in GeneXpert-instruments hasmore potential to detect of positive MRSA samples compared to Nasal Complete-kit and inhousePCR.Conclusion: NxG-kit could easily detect the presence of MRSA because the kit targetsSCCmec carrying mecA- or mecC-gene in the orfX-gene, which provides accurate and rapidMRSA detection.

  • 31.
    Backlund, Ingrid
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of a selective media for the detection of gram-positive bacteria in leg ulcers and pressure wounds2015Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Hard-to-heal ulcers are resource intensive due to the fact that they are difficult to treat and especially vulnerable to bacterial invasion. The bacterial culture contaminating these wounds often consist of several different bacterial organisms that originate from endogenous sources. Necrotic material in ischemic ulcers provide nutrition which support bacterial reproduction, increasing the risk of infection. Determining causative pathogen in infected ulcers proves to be difficult when culturing swab samples, however Staphylococcus aureus and hemolytic streptococci generally act as primary pathogens.

        The aim of the study was to investigate if the detection rate increased for S. aureus and hemolytic streptococci when culturing swab samples from ulcers on Columbia CNA; a media selective for gram-positive bacteria. In the experimental procedure the inhibitory action of CNA upon gram-negative bacterial growth was evaluated, using simulated ulcer samples (n=6) containing bacterial quality control strains in arbitrary concentrations. Additionally, patient samples (n=51) were cultured and screened for primary pathogens to investigate differences in the detection rate for CNA and the current culture media; Blood agar, Chocolate agar, Gentian violet blood agar and CLED agar.

       Results from simulated ulcer samples showed excellent inhibitory function regarding the antibiotic substances of the CNA agar. Culturing patient samples from lower leg- and pressure ulcers on CNA, provided indications of diverse circumstances yielding higher respectively lower detection rate concerning S. aureus and hemolytic streptococci. Samples containing mixed flora with gram-negative bacteria generated higher detection rate and samples containing S. aureus yielded a lower detection rate when culturing on CNA, compared with that of the routine method. 

  • 32. Bahabozorgi, Bahareh
    Optimization of Legionella diagnostics by developing a multiplex real-time PCR assay for simultaneous detection of Legionella pneumophila and Legionella species2017Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    The genus Legionella are intracellular organisms causing infection in the lower respiratory tracts and are accountable for global outbreaks of Legionnaires' disease. Legionella exists in various aquatic environments i.e. water systems in hospital buildings. Immunosuppressed patients are particularly vulnerable, considering the relatively high mortality rate within hospital-acquired pneumonia (5-20 %). The aim of this study was to optimize the current Legionella diagnostics by comparing different extraction methods and three real-time PCR assays for detecting L. species and L. pneumophila. Material used in this study consisted of nine clinical specimens collected from the respiratory tract from inpatients at the Uppsala University Hospital and surrounding regions, ten clinical samples from Quality Control for Molecular Diagnostics and reference bacterial cultures from Culture Collection University of Gothenburg. Extraction methods were tested with different pretreatment procedures. Different concentrations of primers and probes were combined to optimize the sensitivity and specificity of the method. Phocine herpesvirus-1 served as an inhibition control within the multiplex PCR assay. The results demonstrated 90 % and 100 % agreement when comparing current method with the commercial kit and the multiplex PCR assay, respectively. Sensitivity and specificity were improved using primer concentrations above 0.5 μM with 0.2 μM probe. Using Phocine herpesvirus-1 as an inhibition control showed successful detection in the multiplex PCR assay, although it had an impact on the detection of L. pneumophila and L. species. This indicates competition of reagents within the reaction, thus further optimization is required to improve the multiplex PCR assay.

  • 33.
    Bahrmann, Philipp
    et al.
    Friedrich Alexander Univ, Inst Biomed Aging.
    Bertsch, Thomas
    Paracelsus Med Univ, Gen Hosp Nuremberg, Inst Clin Chem Lab Med & Transfus Med.
    Giannitsis, Evangelos
    Univ Hosp Heidelberg, Dept Cardiol.
    Christ, Michael
    Luzerner Kantonsspital, Emergency Dept.
    Hofner, Benjamin
    Friedrich Alexander Univ, Dept Med Informat Biometry & Epidemiol.
    Christenson, Robert
    Univ Maryland, Sch Med, Dept Pathol.
    Lindahl, Bertil
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Cardiology. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Medicinska och farmaceutiska vetenskapsområdet, centrumbildningar mm, UCR-Uppsala Clinical Research Center.
    Mueller, Christian
    Univ Hosp Basel, Dept Cardio.;Univ Hosp Basel, Cardiovasc Res Inst Basel.
    Quantification of Renal Function and Cardiovascular Mortality in Patients Admitted to the Emergency Department with Suspected Acute Coronary Syndromes: Results from the TRAPID-AMI Study2017In: Clinical Laboratory, ISSN 1433-6510, Vol. 63, no 9, p. 1457-1466Article in journal (Refereed)
    Abstract [en]

    Background: Increases in the novel serum marker cystatin C are detectable much earlier in the course of chronic kidney disease (CKD) even when levels of serum creatinine are still in the normal range. A major factor causing a decrease in serum creatinine is increasing age. Patients with CKD are more likely to develop cardiovascular disease (CVD) than a healthy population and to suffer premature deaths from CVD related to CKD. The aim of this study was to investigate whether cystatin C, serum creatinine, and estimated glomerular filtration rate (eGFR) predict cardiovascular mortality in patients admitted to the emergency department (ED) with suspected acute coronary syndromes (ACS).

    Methods: In 1,282 patients (mean age 62 15 years, 477 women, 805 men) with suspected ACS, baseline cystatin C concentrations, serum creatinine, and estimated glomerular filtration rate (eGFR) were measured at the ED. Clinical assessment and serial high sensitivity cardiac troponin T (hs-cTnT) measurements were used for the diagnosis of ACS. Seventeen cardiovascular deaths were registered during a median follow-up of 365 days.

    Results: HRs from univariate Cox regression models for each of the potential biomarkers were 12.02 (95% CI 5.10 - 28.34) for cystatin C, 4.53 (1.75 - 11.70) for serum creatinine, and 0.97 (0.96 - 0.99) for eGFR. All three biomarkers showed a significant association with cardiovascular mortality in univariate analyses. The HRs from a model with all three potential biomarkers were 59.21 (95% CI 9.69 - 361.76) for cystatin C, 0.08 (0.01 - 0.58) for serum creatinine, and 0.98 (0.96 - 1.01) for eGFR. The risk association was significant for ln (cystatin C) and ln (serum creatinine).

    Conclusions: Results of this prospective study show that the quantification of renal function using cystatin C is useful for predicting cardiovascular mortality in patients with suspected ACS at the ED.

  • 34.
    Bentley, Katie
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Vascular Biology. Harvard Med Sch, Beth Israel Deaconess Med Ctr, Computat Biol Lab, Boston, MA USA..
    Chakravartula, Shilpa
    Harvard Med Sch, Beth Israel Deaconess Med Ctr, Computat Biol Lab, Boston, MA USA..
    The temporal basis of angiogenesis2017In: Philosophical Transactions of the Royal Society of London. Biological Sciences, ISSN 0962-8436, E-ISSN 1471-2970, Vol. 372, no 1720, p. 1-11, article id 20150522Article in journal (Refereed)
    Abstract [en]

    The process of new blood vessel growth (angiogenesis) is highly dynamic, involving complex coordination of multiple cell types. Though the process must carefully unfold over time to generate functional, well-adapted branching networks, we seldom hear about the time-based properties of angiogenesis, despite timing being central to other areas of biology. Here, we present a novel, time-based formulation of endothelial cell behaviour during angiogenesis and discuss a flurry of our recent, integrated in silico/in vivo studies, put in context to the wider literature, which demonstrate that tissue conditions can locally adapt the timing of collective cell behaviours/decisions to grow different vascular network architectures. A growing array of seemingly unrelated 'temporal regulators' have recently been uncovered, including tissue derived factors (e.g. semaphorins or the high levels of VEGF found in cancer) and cellular processes (e.g. asymmetric cell division or filopodia extension) that act to alter the speed of cellular decisions to migrate. We will argue that 'temporal adaptation' provides a novel account of organ/disease-specific vascular morphology and reveals 'timing' as a new target for therapeutics. We therefore propose and explain a conceptual shift towards a 'temporal adaptation' perspective in vascular biology, and indeed other areas of biology where timing remains elusive. This article is part of the themed issue 'Systems morphodynamics: understanding the development of tissue hardware'.

  • 35.
    Bergfors, Monica
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of Microsatellite Instability Analysis as a Diagnostic Tool to Identify Lynch Syndrome in Endometrial Cancer Patients2014Independent thesis Advanced level (degree of Master (One Year)), 20 credits / 30 HE creditsStudent thesis
    Abstract [en]

    Hereditary endometrial cancer (EC) is a Lynch syndrome (LS) related cancer variant and 2-10% of all EC are hereditary. The aim of this study was to develop a method for analysis of microsatellite instability (MSI) as such analysis would assist in identifying potential LS patients with EC at an early state of their disease, before a possible second cancer is developed in another organ.

    Twenty-six patients with adenocarcinoma in the endometrium, diagnosed at Uppsala University Hospital in Sweden between 1993 and 2012, were included in the study. Seven of these patients were also diagnosed with LS and the rest were sporadic EC. DNA was extracted from the patients’ formalin-fixed and paraffin-embedded tissues. The extracted DNA was subjected to a multiplex PCR with fluorescently labelled primers and then analysed by using capillary electrophoresis.

    Of the sporadic EC, 26% was MSI-High, which correlates well with published data. Of the LS patients, 83% was MSI-High. The outcome of this project resulted in that MSI analysis is now a validated and established method used in the process of identifying potential LS among patients with EC.

  • 36. Berglund, Elias
    The effects of probiotics on sleep and fatty acids2014Independent thesis Basic level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Probiotics are dietary supplements that contain bacteria that are potentially beneficial for the intestinal flora. The positive effects of probiotics are however not limited to the intestine. The use of probiotics is becoming more and more common and therefore needs to be studied more closely.The purpose of this study is to see how healthy individuals, in the age between 18 and 28 years old and with a BMI between 20 and 25, respond to the probiotic LactiPlus (also called FF8).Blood samples were collected before and after the subjects have eaten a standardized meal. The subject´s glucometabolic responses to food, sleep patterns and their fatty acid profile was analyzed in relation to the probiotic composition.Due to difficulties including study subjects, three subject completed the participation in the study. The three study subjects had similar sleeping habits, one had slightly higher fruit intake, the word and number memory were similar, but it was not possible to relate any data to the use of probiotics. It can be summarized that inclusion additional study subjects is needed.

  • 37.
    Bin Kaderi, Mohamed Arifin
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Genetics and Pathology, Hematology and Immunology.
    Assessment of Novel Molecular Prognostic Markers in Chronic Lymphocytic Leukemia2010Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The clinical course of chronic lymphocytic leukemia (CLL) is highly heterogeneous, which has prompted the search for biomarkers that can predict prognosis in this disease. The IGHV gene mutation status and certain genomic aberrations have been identified as reliable prognostic markers of clinical outcome for this disorder. However, the search for more feasible prognostic markers in CLL is still being pursued. Recently, certain single nucleotide polymorphisms (SNPs) in the GNAS1, BCL2 and MDM2 genes and the RNA expression levels of the LPL, ZAP70, TCL1, CLLU1 and MCL1 genes were suggested as novel prognostic markers in CLL.

    In papers I-III, we performed genotyping analyses of the GNAS1 T393C, BCL2 -938C>A and MDM2 SNP309 polymorphisms in 268-418 CLL patients and related the genotypes with clinical data. Association studies between the polymorphisms and established prognostic markers (i.e. IGHV mutation status, genomic aberrations, CD38 expression) were also performed. Our studies did not find any significant relationship between these SNPs with either clinical outcome or other known prognostic markers in CLL.

    In paper IV, we measured the RNA expression levels of LPL, ZAP70, TCL1, CLLU1 and MCL1 in 252 CLL cases and correlated these levels with clinical outcome. Here, we verified that high expression of all these RNA-based markers, except MCL1, were associated with an unfavourable prognosis. We also confirmed a close relationship between IGHV mutation status and the RNA-based markers, especially for LPL and CLLU1 expression. Among the RNA-based markers, multivariate analysis revealed LPL expression as the strongest independent prognostic marker for overall survival and time to treatment. Furthermore, the RNA-based markers could add further prognostic information to established markers in subgroups of patients, with LPL expression status giving the most significant results.

    In summary, data from papers I-III could not verify the GNAS1 T393C, BCL2 -938C>A and MDM2 SNP309 polymorphisms as prognostic markers in CLL. Future SNP markers must hence be confirmed in large, independent cohorts before being proposed as prognostic marker in CLL. In paper IV, we conclude that LPL expression appears to be the strongest among the RNA-based markers for CLL prognostication. Further efforts to standardize LPL quantification are required before it can be applied in the clinical laboratory to predict clinical outcome in this disease.

    List of papers
    1. The GNAS1 T393C polymorphism and lack of clinical prognostic value in chronic lymphocytic leukemia
    Open this publication in new window or tab >>The GNAS1 T393C polymorphism and lack of clinical prognostic value in chronic lymphocytic leukemia
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    2008 (English)In: Leukemia research: a Forum for Studies on Leukemia and Normal Hemopoiesis, ISSN 0145-2126, E-ISSN 1873-5835, Vol. 32, no 6, p. 984-987Article in journal (Refereed) Published
    Abstract [en]

    Chronic lymphocytic leukemia (CLL) is a clinically heterogeneous disease with no known single predisposing genetic factor shown in all cases. Recently, a single nucleotide polymorphism (SNP) T393C in the GNAS1 gene has been reported to have a clinical impact on CLL progression and overall survival. In order to further investigate the T393C SNP in CLL, we have genotyped 279 CLL cases and correlated the genotypes to clinical outcome and other known prognostic factors such as the immunoglobulin heavy chain variable (IGHV) gene mutation status and CD38 expression. In the present study, no difference in overall survival or time to treatment was observed in the CLL patients with the different genotypes in contrast to the previous report. Furthermore, no correlation was observed with the T393C genotypes and IGHV mutational status, Binet stage or CD38 in this cohort. In summary, our data does not support the use of the T393C GNAS SNP as a clinical prognostic factor in CLL.

    Keywords
    GNAS1 T393C single nucleotide polymorphism, chronic lymphocytic leukemia prognosis
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-13005 (URN)10.1016/j.leukres.2007.10.003 (DOI)000255269100021 ()18006055 (PubMedID)
    Available from: 2008-01-20 Created: 2008-01-20 Last updated: 2017-12-11Bibliographically approved
    2. The BCL-2 promoter (-938C>A) polymorphism does not predict clinical outcome in chronic lymphocytic leukemia
    Open this publication in new window or tab >>The BCL-2 promoter (-938C>A) polymorphism does not predict clinical outcome in chronic lymphocytic leukemia
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    2008 (English)In: Leukemia, ISSN 0887-6924, E-ISSN 1476-5551, Vol. 22, no 2, p. 339-343Article in journal (Refereed) Published
    Abstract [en]

    The (-938C>A) polymorphism in the promoter region of the BCL-2 gene was recently associated with inferior time to treatment and overall survival in B-cell chronic lymphocytic leukemia (CLL) patients displaying the -938A/A genotype and may thus serve as an unfavorable genetic marker in CLL. Furthermore, the -938A/A genotype was associated with increased expression of Bcl-2. To investigate this further, we analyzed the -938 genotypes of the BCL-2 gene in 268 CLL patients and correlated data with treatment status, overall survival and known prognostic factors, for example, Binet stage, immunoglobulin heavy-chain variable (IGHV) mutational status and CD38 expression. In contrast to the recent report, the current cohort of CLL patients showed no differences either in time to treatment or overall survival in relation to usage of a particular genotype. In addition, no correlation was evident between the (-938C>A) genotypes and IGHV mutational status, Binet stage or CD38. Furthermore, the polymorphism did not appear to affect the Bcl-2 expression at the RNA level. Taken together, our data do not support the use of the (-938C>A) BCL-2 polymorphism as a prognostic marker in CLL and argue against its postulated role in modulating Bcl-2 levels.

    Keywords
    chronic lymphocytic leukemia, BCL-2 promoter polymorphism, immunoglobulin heavy-chain variable gene mutation status, Binet stage, CD38, prognosis
    National Category
    Medical and Health Sciences
    Identifiers
    urn:nbn:se:uu:diva-13004 (URN)10.1038/sj.leu.2405042 (DOI)000253166900015 ()18046447 (PubMedID)
    Available from: 2008-01-20 Created: 2009-01-12 Last updated: 2017-12-11Bibliographically approved
    3. Lack of association between the MDM2 promoter polymorphism SNP309 and clinical outcome in chronic lymphocytic leukemia
    Open this publication in new window or tab >>Lack of association between the MDM2 promoter polymorphism SNP309 and clinical outcome in chronic lymphocytic leukemia
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    2010 (English)In: Leukemia research: a Forum for Studies on Leukemia and Normal Hemopoiesis, ISSN 0145-2126, E-ISSN 1873-5835, Vol. 34, no 3, p. 335-339Article in journal (Refereed) Published
    Abstract [en]

    The 309T>G polymorphism in the promoter region of the MDM2gene, known as SNP309, has recently been suggested as an unfavorable prognostic marker in chronic lymphocytic leukemia (CLL) although this has been questioned. To investigate this further, we analyzed the MDM2 SNP309 genotypes in 418 CLL patients and correlated the results with established CLL prognostic factors, time to treatment and overall survival. In this Swedish cohort, no association existed between any particular MDM2 SNP309 genotype, overall survival and time to treatment. Furthermore, no correlation was shown between the MDM2 SNP309 genotypes and Binet stage, IGHV mutational status and recurrent genomic aberrations. In summary, this study argues against the use of the MDM2 SNP309 as a prognostic marker in CLL.

    Keywords
    MDM2 SNP309, Chronic lymphocytic leukemia, Binet stage, IGHV mutational status, Genomic aberrations, Prognostic markers
    National Category
    Medical and Health Sciences
    Research subject
    Clinical Genetics; Medicine; Oncology; Medical Genetics; Molecular Genetics
    Identifiers
    urn:nbn:se:uu:diva-111075 (URN)10.1016/j.leukres.2009.06.006 (DOI)000274529600013 ()19573916 (PubMedID)
    Available from: 2009-12-02 Created: 2009-12-02 Last updated: 2017-12-12Bibliographically approved
    4. LPL is the strongest prognostic factor in a comparative analysis of RNA-based markers in early chronic lymphocytic leukemia
    Open this publication in new window or tab >>LPL is the strongest prognostic factor in a comparative analysis of RNA-based markers in early chronic lymphocytic leukemia
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    2011 (English)In: Haematologica (online), ISSN 0390-6078, E-ISSN 1592-8721, Vol. 96, no 8, p. 1153-1160Article in journal (Refereed) Published
    Abstract [en]

    BACKGROUND:

    The expression levels of LPL, ZAP70, TCL1A, CLLU1 and MCL1 have recently been proposed as prognostic factors in chronic lymphocytic leukemia. However, few studies have systematically compared these different RNA-based markers.

    DESIGN AND METHODS:

    Using real-time quantitative PCR, we measured the mRNA expression levels of these genes in unsorted samples from 252 newly diagnosed chronic lymphocytic leukemia patients and correlated our data with established prognostic markers (for example Binet stage, CD38, IGHV gene mutational status and genomic aberrations) and clinical outcome.

    RESULTS:

    High expression levels of all RNA-based markers, except MCL1, predicted shorter overall survival and time to treatment, with LPL being the most significant. In multivariate analysis including the RNA-based markers, LPL expression was the only independent prognostic marker for overall survival and time to treatment. When studying LPL expression and the established markers, LPL expression retained its independent prognostic strength for overall survival. All of the RNA-based markers, albeit with varying ability, added prognostic information to established markers, with LPL expression giving the most significant results. Notably, high LPL expression predicted a worse outcome in good-prognosis subgroups, such as patients with mutated IGHV genes, Binet stage A, CD38 negativity or favorable cytogenetics. In particular, the combination of LPL expression and CD38 could further stratify Binet stage A patients.

    CONCLUSIONS:

    LPL expression is the strongest RNA-based prognostic marker in chronic lymphocytic leukemia that could potentially be applied to predict outcome in the clinical setting, particularly in the large group of patients with favorable prognosis.

    National Category
    Medical and Health Sciences Medical Genetics Cancer and Oncology
    Research subject
    Clinical Genetics; Medical Genetics; Oncology
    Identifiers
    urn:nbn:se:uu:diva-111078 (URN)10.3324/haematol.2010.039396 (DOI)000294722700013 ()21508119 (PubMedID)
    Available from: 2009-12-02 Created: 2009-12-02 Last updated: 2018-01-12Bibliographically approved
  • 38.
    Björklund, Kristofer
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Evaluation and optimization of four real-time PCRs, using TaqMan-probes, for detection of and discrimination between barley, oat, rye and wheat2008Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Coeliac disease is a chronic inflammatory disease treated with a gluten-free diet, excluding barley, rye and wheat. Hence, there is a demand for methods able to detect gluten in foods in order to ensure correct labeling of products. According to the Codex Alimentarius Commission, 20ppm gluten is the maximum amount allowed in food labeled gluten-free.

    PCR can detect DNA from cereals in food. Four real-time PCR-systems,

    using TaqMan®-probes for detection of barley, oat, rye and wheat were optimized and evaluated. Evaluations were carried out using seeds. Primers were targeted to genes coding for prolamines, seed storage proteins. PCR-systems targeted to barley, oat and wheat were shown to be specific for the cereals corresponding to each system. The system targeted to rye showed cross-reactions with durum wheat and spelt wheat. Detection limits were 50pg, corresponding to <10 haploid genome copies for each cereal. All systems were able to detect 250ppm amounts of DNA, most likely even smaller amounts are detectable. All systems showed an amplification efficiency of ≥95%.

    Systems for detection of barley, oat and wheat are ready for further evaluation, using food products as samples. The rye system however, needs to be re-designed before further evaluation can take place.

  • 39.
    Blom, Kristin
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences.
    Nygren, Peter
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Experimental and Clinical Oncology.
    Larsson, Rolf
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Cancer Pharmacology and Computational Medicine.
    Andersson, Claes R.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Cancer Pharmacology and Computational Medicine.
    Predictive Value of Ex Vivo Chemosensitivity Assays for Individualized Cancer Chemotherapy: A Meta-Analysis2017In: SLAS TECHNOLOGY, ISSN 2472-6303, Vol. 22, no 3, p. 306-314Article in journal (Refereed)
    Abstract [en]

    Current treatment strategies for chemotherapy of cancer patients were developed to benefit groups of patients with similar clinical characteristics. In practice, response is very heterogeneous between individual patients within these groups. Precision medicine can be viewed as the development toward a more fine-grained treatment stratification than what is currently in use. Cell-based drug sensitivity testing is one of several options for individualized cancer treatment available today, although it has not yet reached widespread clinical use. We present an up-to-date literature meta-analysis on the predictive value of ex vivo chemosensitivity assays for individualized cancer chemotherapy and discuss their current clinical value and possible future developments.

  • 40.
    Bollen, Lise Svendsen
    Uppsala University, Department of Comparative Medicine.
    Production of polyclonal antibodies in rabbits and chickens immunised with human immunoglobulin G: A study of the utility of egg yolk antibody production as a substitute for rabbit serum antibody production1997Doctoral thesis, comprehensive summary (Other academic)
    Abstract [en]

    The utility of chicken egg yolk antibodies as an alternative to rabbit antisera was studied by comparing antibody and avidity development in immunised animals (15 rabbits and 30 chickens) during a one-year immunisation scheme. Human IgG was used as the model antigen and the efficacy of three adjuvants, Freund's Complete Adjuvant, Freund's Incomplete Adjuvant and Hunter's TiterMax, was compared. Purification procedures for egg yolk antibodies were developed, rendering the harvest and processing time for yolk and serum antibodies comparable. A majoradvantage of producing egg yolk antibodies instead of rabbit antisera is an increased productivity.Although the antibody response was found to be higher in rabbit serum than in egg yolk of chickensimmunised using identical schemes, the volumes of obtainable antibody source were ten timeshigher with egg yolk than with rabbit sera. Depending on the immunisation scheme and, inparticular, the choice of adjuvant, approximately five times more antibody can be produced per yearby a chicken than by a rabbit. Considering the cost of purchase and maintenance, rabbit antibodiesare ten times more expensive to produce. Serum antibody response in young and old chickens werecompared and no significant difference was found. The avidity of the egg yolk antibodies was foundto be similar to rabbit serum antibodies, but the qualitative properties of the immunoglobulinsdiffer. Different immunochemical and immunoelectrophoretic assays (different ELISAs, liquidphase immunosorbent assay, rocket-immunoelectrophoresis, fused-rocket-immunoelectrophoresis,line-immunoelectrophoresis, crossed-immunoelectro-phoresis, crossed-tandem-immunoelectro-phoresis, crossed-affino-immunoelectrophoresis, charge-shift-crossed-immunoelectrophoresis) weredeveloped for measuring antibody response, and analysing specificity and binding properties. TheIgG levels in developing oozytes (6 - 37 mm) were similar, and there was a significant linearcorrelation between antibody response in serum and corresponding egg yolk. From an animalwelfare point of view there are improvements associated with producing egg yolk antibodies insteadof rabbit serum antibodies.

  • 41.
    Borén, Therese
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Study of proteins related to the thyroid in the endometrium of women with unexplained infertilityTherese Borén2014Independent thesis Basic level (university diploma), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Infertility is a global health issue that can have many causes. It is, however, not always known why some couples cannot conceive even though there are no apparent reason in either the man or the woman. The woman can have normal menstrual cycles and the man can have normal semen.The purpose of this study was to investigate the presence and distribution of thyroid related proteins in endometrial biopsies. Eight biopsies was taken from women with unexplained infertility and 35 biopsies from healthy fertile woman. These were divided into smaller groups based on menstrual cycle day and analysed with immunohistochemistry for the detection of the thyroid related proteins MCT8 and DIO2.There were no significant differences between the groups and therefore are more studies required in the subject to be able to find factors that may help solving some cases of unexplainedinfertility.

  • 42.
    Bouro Wallgren, Sofia
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Tolerance to virus infections could explain increased winter colony survival observed in Varroa destructor-resistant honey bees2018Independent thesis Advanced level (professional degree), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Honey bee colonies all over Europe and North America have been declining dramatically for over three decades and is continuing to do so which is causing significant threats to economy, agriculture and ecosystems. The main reason behind the declining colonies is an ectoparasitic mite known as Varroa destructor and viruses vectored by the mite. In previous studies, it has been suggested that a unique mite-resistant subpopulation of honey bees (Apis mellifera) in Gotland, Sweden have developed adaptive tolerance to these viruses as they have managed to survive high mite infestation through natural selection without any mite control treatment. This indicates that there might be a correlation between resistance to Varroa destructor and virus tolerance. This project examined if a correlation between virus resistance and/or virus tolerance can be observed in Varroa-resistant honey bees from unique subpopulations in Europe covering Sweden, Norway, France and Netherlands. Results showed that no correlation could be established based on the findings in this project. However, significant differences in winter colony survival numbers between mite-resistant and mite-susceptible honey bees suggest that tolerance mechanisms could be present in these subpopulations. Further studies are required to verify this hypothesis.

  • 43.
    Broddesson, Sandra
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Evaluation of an automated multiplex real-time RT-PCR assay for rapid detection of Influenza A and B viruses2015Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Influenza is a viral infection that affects global health and economy with its endemic and sometimes pandemic spread. Rapid detection of Influenza viruses enables antiviral use and can bring financial savings. It is also essential for the global surveillance of prevalent Influenza strains. RT-PCR is considered the most specific and sensitive method for detection of Influenza, but Influenza mutates at a high rate and it is therefore crucial that RT-PCR methods are updated regularly.

    In 2014, Cepheid released their Xpert Flu/RSV XC assay, which can detect Influenza A and B and RSV by multiplex RT-PCR in approximately one hour. The aim of this study was to evaluate this assay at Laboratoriemedicin Västernorrland by using the laboratory’s previous PCR assay for detection of Influenza viruses as reference method.

    Real-time RT-PCR was used to compare Xpert Flu/RSV XC to the reference method. A dilution series was performed to estimate the methods’ PCR efficiencies and precision was calculated from quadruplicates of a positive control sample. Clinical specimens (n=42) were used to evaluate the diagnostic sensitivity and specificity of Xpert Flu/RSV XC. Objective statistical analysis of PCR data was performed and discussed.

    The Xpert Flu/RSV XC was equivalent to the reference method and demonstrated high diagnostic sensitivity and specificity. Estimated PCR efficiencies were however low.

    With the introduction of Xpert Flu/RSV XC to the laboratory follows many potential benefits, primarily in form of a simplified pre analytical procedure and a shortened analysis time. The Xpert Flu/RSV XC assay enables fast diagnosis of Influenza infection.

  • 44.
    Broman, Mikael
    et al.
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Cell Biology.
    Frenckner, Bjorn
    Bjallmark, Anna
    Broome, Michael
    Recirculation during veno-venous extra-corporeal membrane oxygenation - a simulation study2015In: International Journal of Artificial Organs, ISSN 0391-3988, E-ISSN 1724-6040, Vol. 38, no 1, p. 23-30Article in journal (Refereed)
    Abstract [en]

    Purpose: Veno-venous ECMO is indicated in reversible life-threatening respiratory failure without life-threatening circulatory failure. Recirculation of oxygenated blood in the ECMO circuit decreases efficiency of patient oxygen delivery but is difficult to measure. We seek to identify and quantify some of the factors responsible for recirculation in a simulation model and compare with clinical data. Methods: A closed-loop real-time simulation model of the cardiovascular system has been developed. ECMO is simulated with a fixed flow pump 0 to 5 l/min with various cannulation sites -1) right atrium to inferior vena cava, 2) inferior vena cava to right atrium, and 3) superior+ inferior vena cava to right atrium. Simulations are compared to data from a retrospective cohort of 11 consecutive adult veno-venous ECMO patients in our department. Results: Recirculation increases with increasing ECMO-flow, decreases with increasing cardiac output, and is highly dependent on choice of cannulation sites. A more peripheral drainage site decreases recirculation substantially. Conclusions: Simulations suggest that recirculation is a significant clinical problem in veno-venous ECMO in agreement with clinical data. Due to the difficulties in measuring recirculation and interpretation of the venous oxygen saturation in the ECMO drainage blood, flow settings and cannula positioning should rather be optimized with help of arterial oxygenation parameters. Simulation may be useful in quantification and understanding of recirculation in VV-ECMO.

  • 45.
    Bäckryd, Emmanuel
    et al.
    Linköping University, Linköping, Sweden.
    Lind, Anne-Li
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences.
    Thulin, Måns
    Uppsala University, Disciplinary Domain of Humanities and Social Sciences, Faculty of Social Sciences, Department of Statistics.
    Larsson, Anders
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences.
    Gerdle, Björn
    Linköping University, Linköping, Sweden.
    Gordh, Torsten
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Surgical Sciences.
    High Levels of Cerebrospinal Fluid Chemokines Point to the Presence of Neuroinflammation in Peripheral Neuropathic Pain: A Cross-Sectional Study of Two Cohorts of Patients Compared to Healthy ControlsManuscript (preprint) (Other academic)
    Abstract [en]

    Animal models suggest that chemokines are important mediators in the pathophysiology of neuropathic pain. Indeed, these substances have been called “gliotransmitters”, a term that illustrates the close interplay between glial cells and neurons in the context of neuroinflammation and pain. However, evidence in humans is scarce. The aim of the study was to determine a comprehensive cerebrospinal fluid (CSF) inflammatory profile for neuropathic pain patients. Our hypothesis was that we would thereby find indications of a postulated on-going process of central neuroinflammation.  

    CSF samples were collected from two cohorts of patients with neuropathic pain (n=11 and n=16, respectively) and healthy controls (n=11). The samples were analyzed with a multiplex proximity extension assay in which 92 inflammation-related proteins were measured simultaneously (Proseek® Multiplex Inflammation I, Olink Bioscience, Uppsala, Sweden). Univariate testing with control of false discovery rate, as well as orthogonal partial least squares – discriminant analysis, were used for statistical analyses.

    CSF levels of chemokines CXCL6, CXCL10, CCL8, CCL11, CCL23, as well as protein LAPTGF-beta-1, were significantly higher in both neuropathic pain cohorts compared to healthy controls, pointing to neuroinflammation in patients. These 6 proteins were also major results in a recent similar study in fibromyalgia patients. The findings need to be confirmed in larger cohorts, and the question of causality remains to be settled. Since it has been suggested that prevalent co-morbidities to chronic pain (e.g., depression, anxiety, poor sleep, and tiredness) also are associated with inflammation, it will be important to determine whether inflammation is a common mediator.

  • 46.
    Camacho, Emely
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Optimization of Lentivirus Production for Cancer Therapy2011Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Vectors based on lentivirus backbones have revolutionized our ability to transfer genesinto many cell types. Lentiviral vectors integrate into the chromatin of target cells and do not transfer any viral genes causing vector replication. Both of these features arecommonly used in gene therapy and have been used clinically in individuals sufferingfrom cancer, infections and genetic diseases. It has been discovered that T-cells can be genetically modified to be used as effective weapons against cancer: therefore virus mustbe produced to deliver the gene of interest into the T-cells. In this project, lentiviralvectors have been produced to transfer the gene coding for a chimeric antigen receptor(CAR) which is directed to CD19 on B-cells. The vectors will, hence, be used to generateCD19 retargeted T-cells in purpose to kill CD19 cells such as B-cell lymphoma andleukemia. We have evaluated two production protocols to determine a feasible method toculture these vectors. We have also stimulate T-cells with two different antibodies (anti-CD3 and anti-CD28) and transduced T-cells. Our results demonstrate that theconcentration of virus was higher after prolonged incubation in 4˚C, which can not beexplained. The stimulation demonstrated that bound anti-CD3 was the best stimulator,and moreover the FACS-analysis showed that addition of anti-CD28 gave a highertransduction level. In conclusion, the viral vectors may be kept in 4˚C for two days beforeconcentrating the virus, and bound anti-CD3 is a better choice than soluble anti-CD3 forstimulation of T-cells.

  • 47.
    Carls, Stefan
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Optimization of pyrosequencing method for copy number analysis of CYP2D62017Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    CYP2D6, a member of the cytochrome P450 enzyme system, has a central role in drug metabolism, it metabolizes 25 % of clinically used drugs. The gene that codes for the enzyme displays a high degree of polymorphism, which effects enzyme functions to various degrees. Aside from smaller mutations like SNPs, alleles may also feature duplications or deletion of the whole gene. Due to the clinical relevance of these mutations, a simple and precise method for genotyping is needed. In this study, a method based on pyrosequencing for copy number analysis was evaluated, wherein the copy number was determined by relative quantification to a reference gene CYP2D8P. During evaluation of the method, several adjustments were tried for optimization, including adjustments of annealing temperature and primer concentration. The results showed a difficulty in distinguishing between copy numbers using the method, as well as a high coefficient of variation. Therefore, further optimization is required before the method could be implemented into clinical practice.

  • 48.
    Cedergren, Linda
    Uppsala University, Medicinska vetenskapsområdet, Faculty of Medicine, Department of Medical Biochemistry and Microbiology.
    Expression of recombinant protein including an His-tag to facilitate purification for diagnosis of CCHF and Lassa Viruses2006Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Abstract

    Crimean-Congo Hemorrhagic Fever virus (CCHF) and Lassa virus are giving sources illness to humans. In addition to zoonotic transmission, CCHF and Lassa virus can spread from person to person. After a short incubation period, CCHF and Lassa virus infections are characterized by a sudden onset of high fever, chills, headache and cough just like flu. Even some people are vomiting and have diarrhoea. After a few days of illness hemorrhagic manifestations occur. Treatment options for CCHF and Lassa viruses are limited, and there is no vaccine available for use in humans. The purpose of the present study was to produce recombinant nucleocapsid protein of Lassavirus and CCHF virus including an aminoterminal His-tag by a Semliki Forest Virus Replicon (pSFV 4.2). The recombinant proteins are planned to be used in future development of diagnostic methods.

  • 49.
    Chabo, Maria
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Medical Sciences, Clinical Physiology.
    Ventilation heterogeneity, assessed by nitrogen washout, in relation to Chronic obstructive pulmonary disease symptoms2018Independent thesis Advanced level (professional degree), 10 credits / 15 HE creditsStudent thesis
  • 50.
    Chen, Gunilla
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Women's and Children's Health.
    Changes in adipose tissue mRNA expression due to perinatal exposure to bisphenol A in rats2014Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesis
    Abstract [en]

    Bisphenol A (BPA) is an estrogen receptor binding chemical, widely used in the plastics industry, and as such commonly encountered from plastic containers etc. Even at very low doses, BPA is believed to induce obesity and to have various endocrine disruptive effects. The purpose of this study was to determine possible gene expression changes in gonadal and inguinal adipose tissue from rats perinatally exposed to BPA. The method used was quantitative real-time PCR, and genes found to be up-regulated were PLZF, adiponectin, RXRa and Tcf21, while down-regulated genes were PPARγ, Tmem26, EsR1, Resistin, LPL, Chemerin, Serpina6, TFAM and Ahr. This is so far largely unsupported by other studies, and more research is needed.

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