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Computerised method for recording platelet density distribution
Department of Clinical Chemistry, Örebro Medical Center Hospital, Örebro, Sweden.
1995 (English)In: European Journal of Haematology, ISSN 0902-4441, E-ISSN 1600-0609, Vol. 54, no 5, 304-309 p.Article in journal (Refereed) Published
Abstract [en]

In the present study a computerized apparatus was employed for scanning light transmission variations along test tubes containing density-separated platelets. The device consists of a stepping motor, a stationary halogen lamp and a photopotentiometer connected to a personal computer. Anticoagulated whole blood was layered on a preformed continuous Percoll gradient having a density span from 1090 kg/l (bottom) to 1040 kg/l (top). After centrifugation at 3400g for 1.5 hours, high-density cells (i.e. erythrocytes) pass through to the bottom of the test tube and the lighter platelets remain in the gradient. The test tube is moved by the computer between the halogen lamp and the photopotentiometer. Transmission variations along the gradient were recorded and registered in the computer. Density markers beads were used as an internal standard and platelet peak density was determined. After perforating the test tube the gradient was divided into 45 aliquots. In all fractions determination of platelet counts and mean platelet volume was carried out. In addition, in the aliquots having a platelet count > 20 × 1012/l the ratio β-thromboglobulin per platelet was also determined. The platelet distribution in the gradient was illustrated graphically. A good agreement was found when comparing platelet distributions in the gradients and light transmission variations along the test tubes.

Place, publisher, year, edition, pages
1995. Vol. 54, no 5, 304-309 p.
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:liu:diva-84674DOI: 10.1111/j.1600-0609.1995.tb00690.xOAI: oai:DiVA.org:liu-84674DiVA: diva2:561103
Available from: 2012-10-17 Created: 2012-10-17 Last updated: 2017-12-07Bibliographically approved
In thesis
1. Platelets and the inflammatory response in coronary heart disease
Open this publication in new window or tab >>Platelets and the inflammatory response in coronary heart disease
2003 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The thesis investigates a new invented computerised apparatus suitable for determining platelet density distribution and peak platelet density in undisturbed linear Percoll™ gradients. The device measures light transmission through test tubes containing density-separated platelets. The transmission reflects the distribution of platelets in the gradient. Coloured particles (density marker beads™) with known density are used as calibration. Consequently, the technique makes it possible to determine peak platelet density by comparing the position of the platelet peak in the gradient with the locations of the beads. The thesis describes the construction of the device and investigates its reliability (paper I). A subsequent article (II) compares platelet density alterations and changes of platelet reactivity as estimated from the ADP-evoked platelet fibrinogen binding. We examined individuals with stable angina pectoris (n=18) subject to coronary angiography. Sampling was carried out immediately before angiography and 24 hours thereafter. In a further study (paper III) platelet density was measured in patients having acute myocardial infarctions with ST-segment elevations (STEMI) (n=36). Platelet counts and volumes were also determined. Soluble P-selectin was used to assess platelet activity. The neutrophil and monocyte counts were used to estimate the inflammatory response. Myeloperoxidase and interleukin 6 (IL-6) were employed to quantify neutrophil and monocyte activity, respectively. All measures except platelet density were repeated after 6 months of recovery.

The second part of the thesis (papers IV and V) investigates Chlamydia pneumoniae in coronary heart disease. Paper IV studies a cohort (n=92) with stable angina pectoris. C. pneumoniae IgG was compared with disease severity i.e. the number of diseased coronary arteries (1-3) as determined by coronary angiography. Determination of the following was carried out before angiography: C. pneumoniae IgG, neutrophil count, myeloperoxidase and IL-6. Article V examines if the organism affects platelet activity in acute STEMI (n=I4). In this study C. pneumoniae IgM and IgG together with soluble P-selectin were determined on day I. Except for IgM the measures were repeated after 6 months. Finally, in the last article (paper VI) individual heterogeneity of platelet inhibition after a clopidogrel-loading dose was explored. Platelet reactivity as estimated from the ADP-evoked platelet fibrinogen binding was determined before angiography, stenting and the clopidogrel load (day I). The analysis was repeated after 24 hours (day 2). The thesis demonstrates that the optical apparatus is technically reliable (paper I). Healthy individuals repeatedly have platelet density subpopulations as evidenced from additional transmission peaks (paper I). The subpopulations frequently display enhanced a-granule content as expected from their positions in the gradient (paper I). The second paper shows that platelet density alterations are inversely related to changes of platelet reactivity (paper II). Decreased platelet count and an elevated inflammatory response are features of acute STEMI (paper III). We demonstrate that some patients have substantial inflammatory reactions whereas others had lower inflammatory responses. The difference persisted in the recovery. Compared to subjects with higher platelet density, individuals with lower/disturbed density displayed more severe inflammatory characteristics (paper III). Manuscript IV shows that C. pneumoniae is associated to the severity of coronary atherosclerosis as estimated from to coronary angiography. Paper V reveals that elevated C. pneumoniae IgM at the acute STEMI is related to enhanced soluble P-selectin. Both parameters proved to be associated with IgG both at the MI and after 6 months of recovery (paper V). Thus, enhanced soluble P-selectin at the STEMI is associated with a reactivation of a chronic C. pneumoniae infection. Finally, the thesis shows a substantial individual heterogeneity of platelet inhibition after a clopidogrel load (paper VI). Some individuals had strong inhibition most likely susceptible to bleedings. Others had weak reactions after clopidogrel exposure indicating elevated risk for thrombotic events.

Place, publisher, year, edition, pages
Linköping: Linköpings universitet, 2003. 62 p.
Series
Linköping University Medical Dissertations, ISSN 0345-0082 ; 816
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:liu:diva-27503 (URN)12158 (Local ID)91-7373-505-1 (ISBN)12158 (Archive number)12158 (OAI)
Public defence
2003-10-31, Elsa Brändström salen, Hälsouniversitetet, Linköping, 13:00 (Swedish)
Opponent
Available from: 2009-10-08 Created: 2009-10-08 Last updated: 2012-10-17Bibliographically approved

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Järemo, Petter

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