Allan Carlsson

Cryptococcus neoformans Ex Vivo Capsule Size Is Associated With Intracranial Pressure and Host Immune Response in HIV-associated Cryptococcal Meningitis

Background. The Cryptococcus neoformans polysaccharide capsule is a well-characterized virulence factor with immunomodulatory properties. The organism and/or shed capsule is postulated to raise intracranial pressure (ICP) in cryptococcal meningitis (CM) by mechanical obstruction of cerebrospinal fluid (CSF) outflow. Little is known regarding capsule phenotype in human cryptococcosis. We investigated the relationship of ex vivo CSF capsular phenotype with ICP and CSF immune response, as well as in vitro phenotype. Methods. In total, 134 human immunodeficiency virus (HIV)-infected Ugandan adults with CM had serial lumbar punctures with measurement of CSF opening pressures, quantitative cultures, ex vivo capsule size and shedding, viscosity, and CSF cytokines; 108 had complete data. Induced capsular size and shedding were measured in vitro for 48 C. neoformans isolates. Results. Cryptococcal strains producing larger ex vivo capsules in the baseline (pretreatment) CSF correlated with higher ICP (P = .02), slower rate of fungal clearance (P = .02), and paucity of CSF inflammation, including decreased CSF white blood cell (WBC) count (P < .001), interleukin (IL)-4 (P = .02), IL-6 (P = .01), IL-7 (P = .04), IL-8 (P = .03), and interferon γ (P = .03). CSF capsule shedding did not correlate with ICP. On multivariable analysis, capsule size remained independently associated with ICP. Ex vivo capsular size and shedding did not correlate with that of the same isolates grown in vitro. Conclusions. Cryptococcal capsule size ex vivo is an important contributor to virulence in human cryptococcal meningitis.

Fiber Orientation Control Related to Papermaking

This thesis deals with fibre orientation in wall-bounded shear flows. The primary application in mind is papermaking. The study is mainly experimental,but is complemented with theoretical considerations.The main part of the thesis concerns the orientation of slowly settlingfibres in a wall-bounded viscous shear flow. This is a flow case not dealt withpreviously even at small Reynolds numbers. Experiments were conducted usingdilute suspensions with fibres having aspect ratios of rp ≈ 7 and 30. It is foundthat the wall effect on the orientation is small for distances from the wall wherethe fibre centre is located farther than half a fibre length from the wall. Farfrom the wall most fibres were oriented close to the flow direction. Closer tothe wall than half a fibre length the orientation distribution first shifted to bemore isotropic and in the very proximity of the wall the fibres were orientedclose to perpendicular to the flow direction, nearly aligned with the vorticityaxis. This was most evident for the shorter fibres with rp ≈ 7.Due to the density difference between the fibres and the fluid there is anincreased concentration near the wall. Still, a physical mechanism is requiredin order for a fibre initially oriented close to the flow direction at about half afibre length from the wall to change its orientation to aligned with the vorticityaxis once it has settled down to the wall. A slender body approach is usedin order to estimate the effect of wall reflection and repeated wall contacts onthe fibre rotation. It is found that the both a wall reflection, due to settlingtowards the wall, and contact between the fibre end and the wall are expectedto rotate the fibre closer to the vorticity axis. A qualitative agreement withthe experimental results is found in a numerical study based on the theoreticalestimation.In addition an experimental study on fibre orientation in the boundarylayers of a headbox is reported. The orientation distribution in planes parallelto the wall is studied. The distribution is found to be more anisotropic closerto the wall, i.e. the fibres tend to be oriented closer to the flow direction nearthe wall. This trend is observed sufficiently far upstream in the headbox.Farther downstream no significant change in the orientation distribution couldbe detected for different distances from the wall.

Counterflow Dielectrophoresis for Trypanosome Enrichment and Detection in Blood

Human African trypanosomiasis or sleeping sickness is a deadly disease endemic in sub-Saharan Africa, caused by single-celled protozoan parasites. Although it has been targeted for elimination by 2020, this will only be realized if diagnosis can be improved to enable identification and treatment of afflicted patients. Existing techniques of detection are restricted by their limited field-applicability, sensitivity and capacity for automation. Microfluidic-based technologies offer the potential for highly sensitive automated devices that could achieve detection at the lowest levels of parasitemia and consequently help in the elimination programme. In this work we implement an electrokinetic technique for the separation of trypanosomes from both mouse and human blood. This technique utilises differences in polarisability between the blood cells and trypanosomes to achieve separation through opposed bi-directional movement (cell counterflow). We combine this enrichment technique with an automated image analysis detection algorithm, negating the need for a human operator.

Rheology at the micro-scale: new tools for bio-analysis

We present a simple and non-invasive experimental procedure to measure the linear viscoelastic properties of cells by passive particle tracking microrheology. In order to do this, a generalised Langevin equation is adopted to relate the timedependent thermal fluctuations of a probe sensor, immobilised to the cell’s membrane, to the frequency-dependent viscoelastic moduli of the cell. The method has been validated by measuring the linear viscoelastic response of a soft solid and then applied to cell physiology studies. It is shown that the viscoelastic moduli are related to the cell’s cytoskeletal structure, which in this work is modulated either by inhibiting the actin/myosin-II interactions by means of blebbistatin or by varying the solution osmolarity from iso- to hypo-osmotic conditions. The insights gained from this form of rheological analysis promises to be a valuable addition to physiological studies; e.g. cell physiology during pathology and pharmacological response.

Fibre orientation measurements near a headbox wall

Abstract Experimental results on the fibre orientation in a laboratory scale headbox are reported. Images containing fibres in approximately 1 mm thick slices parallel to the wall were captured at different wall distances. A steerable filter was used to determine the orientation of bleached and unbeaten birch fibres, suspended in water, at different distances from one of the inclined walls of the headbox contraction. Due to optical limitations only dilute suspensions were studied. It is shown that the fibre orientation distribution varies with the distance from the wall. Sufficiently far upstream in the headbox a more anisotropic distribution is found closer to the wall.

Fibre Orientation Control Related to Papermaking

The wall effect on the orientation of fibres suspended in a shear flow has been studied experimentally. A fibre suspension, driven by gravity down an inclined glass plate, constitutes the shear flo ...