Anders Ljungman

Properties and toxicological effects of particles from the interaction between tyres, road pavement and winter traction material

In regions where studded tyres and traction material are used during winter, e.g. the Nordic countries, northern part of USA, Canada, and Japan, mechanically generated particles from traffic are the main reason for high particle mass concentrations in busy street and road environments. In many Nordic municipalities the European environmental quality standard for inhalable particles (PM(10)) is exceeded due to these particles. In this study, particles from the wear of studded and studless friction tyres on two pavements and traction sanding were generated using a road simulator. The particles were characterized using particle sizers, Particle Induced X-Ray Emission Analysis and electron microscopy. Cell studies were conducted on particles sampled from the tests with studded tyres and compared with street environment, diesel exhaust and subway PM(10), respectively. The results show that in the road simulator, where resuspension is minimized, studded tyres produce tens of times more particles than friction tyres. Chemical analysis of the sampled particles shows that the generated wear particles consist almost entirely of minerals from the pavement stone material, but also that Sulfur is enriched for the submicron particles and that Zink is enriched for friction tyres for all particles sizes. The chemical data can be used for source identification and apportionment in urban aerosol studies. A mode of ultra-fine particles was also present and is hypothesised to originate in the tyres. Further, traction material properties affect PM(10) emission. The inflammatory potential of the particles from wear of pavements seems to depend on type of pavement and can be at least as potent as diesel exhaust particles. The results imply that there is a need and a good potential to reduce particle emission from pavement wear and winter time road and street operation by adjusting both studded tyre use as well as pavement and traction material properties.

(1→3)-β-d-Glucan stimulates nitric oxide generation and cytokine mRNA expression in macrophages

Beta-glucans are known for their potent ability to induce nonspecific inflammatory reactions and are believed to play a role in bioaerosol-induced respiratory symptoms seen in both occupational and residential environments. Here, the ability of a (1→3)-β-d-glucan (Curdlan) to stimulate nitric oxide generation and cytokine mRNA expression in rat alveolar macrophages (AMs) and the murine monocyte/macrophage cell line, RAW 264.7 was investigated. Exposure to (1→3)-β-d-glucan (20, 100 and 500 μg/ml) induced a dose-dependent increase in the expression of inducible nitric oxide synthase mRNA and a release of nitric oxide into the culture medium in both rat AMs and RAW 264.7 cells. The mRNA expression of a number of other inflammatory mediators such as interleukin-1β, interleukin-6, tumor necrosis factor-α and cyclooxygenase-2 was also increased by the exposure to β-glucan. The capability of (1→3)-β-d-glucan (500 μg/ml) to induce mRNA synthesis of these various mediators were comparable to that of endotoxin (1 μg/ml). These results imply that (1→3)-β-d-glucan stimulates the generation of nitric oxide, cytokines and prostaglandins in macrophages and suggest the possibility that this may contribute to bioaerosol-induced respiratory symptoms seen in exposed individuals.

Increased Gene Expression of Novel Cytosolic and Secretory Phospholipase A2 Types in Human Airway Epithelial Cells Induced by Tumor Necrosis Factor-α and IFN-γ

Phospholipase A(2) (PLA(2)) is a growing family of enzymes that may play a major role in inflammation. We investigated the effect of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) on the gene expression of 19 different PLA(2) types (IB, IIA, IID, IIE, IIF, III, IVA, IVB, IVC, V, VIA, VIB, VIIA, VIIB, VIIIA, VIIIB, X, XII, and XIII) in human bronchoepithelial (BEAS-2B) and nasal epithelial (RPMI 2650) cells. The cells were stimulated with TNF-alpha or IFN-gamma for different lengths of time (1, 4, 18, and 48 h), and the mRNA levels of the different PLA(2) types were determined by reverse transcriptase-PCR (RT-PCR) and normalized to those of the housekeeping gene, GAPDH. In both cell lines, TNF-alpha increased the expression of PLA(2) IVA and IVC, and IFN-gamma increased the expression of PLA(2) IIA and IID. No influence on the gene expression of PLA(2)-activating protein (PLAP) was noted on cytokine stimulation. These findings indicate that TNF-alpha and IFN-gamma induce gene expression of two novel cytosolic and secretory PLA(2) types (IVC and IID, respectively) in human airway epithelial cells. The possibility that these PLA(2) types are involved in cytokine-mediated inflammation in the respiratory tract is inferred.

Examination involving students as peer examiners

The main interest in this article is students’ involvement in assessment as a part of growth towards self‐directedness in learning. In order to enhance students’ development of autonomy in learning, a project involving ‘older’ students as peer examiners for ‘younger’ students was designed and carried out. Students in the sixth semester in a PBL‐based Master’s program of Medical Biology participated, together with faculty, as examiners of fifth‐semester students. The examination and the assessment situation was carefully designed based on learning theories, empirical evidence and experiences underpinning student‐centred learning, especially in the form of PBL used at the faculty. The project was evaluated and analysed in order to understand students’ learning processes related to the responsibility for assessing peers. The situation of the peer examiners was interpreted based on their own experiences with statements from the students assessed and faculty involved in the assessment. Evaluations from six occasions, spring and fall, 2003–2005, were included in the study. The findings suggest that involving students in assessment as equal partners with faculty makes it is possible for students to apprehend the metacognitive competences needed to be responsible and autonomous in learning. The peer examiners experience motivation to learn about learning, they acquire tacit knowledge about assessment and they learn through being involved and trusted. The student‐centred educational context, which requires responsibility throughout the programme, is recognized as very important.

Expression of members of the phospholipase A2 family of enzymes in human nasal mucosa

Phospholipase A2 (PLA2) is a family of enzymes thought to play a key role in inflammation by releasing arachidonic acid for the synthesis of eicosanoids and lysophospholipid for the synthesis of platelet-activating factor. However, the precise contribution of different PLA2 types to the formation of inflammatory lipid mediators in the upper airways is not known and the expression of different PLA2 genes in the human nasal mucosa has not been examined. This study therefore investigated the occurrence of messenger ribonucleic acids (mRNAs) for different PLA2 forms (IB, IIA, IID, IIE, III, IVA, IVB, IVC, V, VI, VII, X, acid calcium-independent (aiPLA2), and calcium-independent membrane bound PLA2, (iPLA2-2)) in the nasal mucosa of five healthy human subjects. Using reversed transcription-polymerase chain reaction (RT-PCR) techniques it was found that all these PLA2 types except PLA2 V were expressed in all subjects, whereas PLA2 V was detected in only one individual on one single occasion. The relative abundance of the different PLA2 transcripts were aiPLA2 > X approximately = IVA > IIA approximately = IIE approximately = IVB approximately = VI > IB approximately = IID approximately = III approximately = IVC approximately = VII approximately = iPLA2-2. To further quantify the mRNA-expression of PLA2 X, IVA and IIA, the samples were reanalysed with a quantitative PCR-technique utilizing competitive deoxyribonucleic acid (DNA) mimics as references. The amounts of PLA2 X, IVA and IIA mRNA were then estimated to 0.9 +/- 0.2, 1.1 +/- 0.7, and 0.0025 +/- 0.0021 amol (mean +/- SE), respectively, confirming the relative abundance of these PLA2 transcripts and indicating that the recently described PLA2 X form is relatively strongly expressed. These findings demonstrate that a large number of PLA2 types are expressed in the normal human nasal mucosa. Moreover, this investigation demonstrates, for the first time, the presence of the newly discovered phospholipase A2 forms IID, IIE, III, IVB, IVC, X and calcium-independent membrane bound phospholipase A2 in the human nasal mucosa and raises the possibility that one or several of these may be involved in inflammatory reactions in the nose.

Wear Particles from Studded Tires and Granite Pavement Induce Pro-inflammatory Alterations in Human Monocyte-Derived Macrophages : A Proteomic Study.

Airborne particulate matter is considered to be one of the environmental contributors to the mortality in cancer, respiratory, and cardiovascular diseases. For future preventive actions, it is of major concern to investigate the toxicity of defined groups of airborne particles and to clarify their pathways in biological tissues. To expand the knowledge beyond general inflammatory markers, this study examined the toxicoproteomic effects on human monocyte derived macrophages after exposure to wear particles generated from the interface of studded tires and a granite-containing pavement. As comparison, the effect of endotoxin was also investigated. The macrophage proteome was separated using two-dimensional gel electrophoresis. Detected proteins were quantified, and selected proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry. Among analyzed proteins, seven were significantly decreased and three were increased by exposure to wear particles as compared to unexposed control cells. Endotoxin exposure resulted in significant changes in the expression of six proteins: four decreased and two increased. For example, macrophage capping protein was significantly increased after wear particle exposure only, whereas calgizzarin and galectin-3 were increased by both wear particle and endotoxin exposure. Overall, proteins associated with inflammatory response were increased and proteins involved in cellular functions such as redox balance, anti-inflammatory response, and glycolysis were decreased. Investigating the effects of characterized wear particles on human macrophages with a toxicoproteomic approach has shown to be useful in the search for more detailed information about specific pathways and possible biological markers.

CLARA CELL 10-KDA PROTEIN INHIBITS ENDOTOXIN-INDUCED AIRWAY CONTRACTION IN ISOLATED PERFUSED RAT LUNGS

Clara cell 10-kDa protein (CC10) is a major component of bronchoalveolar lavage fluid and is suggested to be a natural regulator of airway inflammation, possibly through its effects on the proinflammatory enzyme(s), phospholipase A 2. We examined the effect of recombinant human (rh) CC10 on endotoxin-induced airway contraction and cytokine release in isolated perfused rat lungs. We found that rhCC10 added to the lung perfusate abolished the endotoxin-induced airway contraction, and that it inhibited both the release of interleukin-1 β and interleukin-6 into the lung perfusate and the release of tumor necrosis factor- α into the pulmonary lavage fluid. By contrast, the levels of interferon- γ were unaffected by CC10 administration. Rutin, a phospholipase A 2 inhibitor, and N ω -nitro- L -arginine methyl ester (L -NAME), a nitric oxide synthase inhibitor, also attenuated the contraction induced by endotoxin. These findings demonstrate that rhCC10 inhibits endotoxin-induced airway contraction and the release of proinflammatory cytokines (interleukin-1 β, interleukin-6, and tumor necrosis factor- α) in isolated perfused rat lungs. The results also indicate that phospholipase A 2 and nitric oxide are involved in the airway contraction in this model, possibly through their influence on the production of eicosanoids.

Calcium Ionophore-Activated Neutrophils Prestimulated with Endotoxin Increase Pulmonary Arterial Pressure and Vascular Leakage in Isolated Perfused Rat Lungs: Role of Platelet-Activating Factor

The influence of stimulated polymorphonuclear neutrophils on pulmonary arterial pressure and vascular leakage in isolated perfused rat lungs was investigated. We exposed isolated neutrophils to various stimuli in vitro, instilled the cells in the lung perfusate, and studied the effects on pulmonary arterial pressure and passage of fluorescently labeled dextran (4100 dalton) from the pulmonary circulation into the lung. We found that neutrophils stimulated with the calcium ionophore A23187 or with E. coli endotoxin had no significant influence on the pressure or the passage of dextran. On the other hand, neutrophils preincubated with endotoxin and then stimulated with A23187 caused significant increases, both in pulmonary arterial pressure and accumulation of dextran in the lung. Both these effects were attenuated by BN 52021, a specific platelet-activating factor antagonist, and by nordihydroguaiaretic acid, an agent that inhibited the generation of platelet-activating factor in A23187-stimulated neutrophils. These findings demonstrate that activated neutrophils can increase pulmonary arterial pressure and lung fluid accumulation and suggest that endotoxin-stimulated activated neutrophils exert at least some of their action via generation of platelet-activating factor.

Two-Dimensional Gel Electrophoresis and Mass Spectrometry in Studies of Nanoparticle-Protein Interactions

Over the years a number of epidemiological studies have shown that PM from combustion sources such as motor vehicles contributes to respiratory and cardiovascular morbidity and mortality.Especially so do the ultra-fine particles (UFPs) with a diameter less than 0.1 micrometer.UFPs from combustion engines are capable to translocate over the alveolar–capillary barrier. When nano-sized PM (nanoparticles, NP), which are small enough to enter the blood stream, do so they are likely to interact with plasma proteins and this protein-NP interaction will probably affect the fate of and the effects caused by the NPs in the human body. Here, by using a proteomic approach, we present results showing that several proteins indeed are associated to NPs that have in vitro been introduced to human blood plasma.

Using a heart operation video with live comments to inspire students in pre-clinical studies - A pilot study

A simple moment of inspiration based on the American empiric psychologist Csikszentmihalyi’s theory about optimal experience, internal motivation and flow was presented to medical students. The stu ...