Main emissive species produced with Helium as plasma carrier gas.

<p>Main emissive species produced with Helium as plasma carrier gas.</p

Electrical high voltage and current signals analyzed during one impulse (2a) and during one period (2b) (He gas flow = 2slm, voltage frequency = 10kHz, voltage amplitude = 5kV, voltage duty cycle = 1).

<p>Electrical high voltage and current signals analyzed during one impulse (2a) and during one period (2b) (He gas flow = 2slm, voltage frequency = 10kHz, voltage amplitude = 5kV, voltage duty cycle = 1).</p

Device settings and cell culture parameters influence cell fate.

<p><b>(a)</b> hPDL were treated with He-GIW using different voltages and <b>(b)</b> exposure lengths. Adherent cell number was evaluated 24h after treatment and expressed as a percentage of control. <b>(c)</b> Cell morphology was monitored by phase contrast microscopy 6h and 24h after treatment. <b>(d)</b> Cell culture incidence on He-GIW treatment was evaluated using different FCS concentrations and <b>(e)</b> cell confluence. Adherent cell number was assayed 24h after treatment and expressed as percentage of control. Error bars represent S.E.M. of three independent experiments p<0.05 (*), p<0.01 (**) and p<0.001 (***).</p

Mitochondrial membrane depolarization precedes cell membrane alteration.

<p><b>(a)</b> Adherent cell percentage was monitored for 6 h after He-GIW treatment. <b>(b)</b> Cell Δψm was monitored by FACS analysis for 6h using DiOC6 staining. Results of one representative experiment of at least three. <b>(c)</b> Geometric means of DiOC6 staining relative to control 1, 3 or 6 h after He-GIW treatment. <b>(d)</b> Δψm (DiOC6) and plasma cell membrane permeability (7AAD) were monitored by FACS analysis for 6 h after He-GIW treatment. When present, error bars represent S.E.M. of three independent experiments p<0.05 (*), p<0.01 (**) and p<0.001 (***)</p

Experimental set-up.

<p>Experimental set-up.</p

Relative concentration of the main emissive species in the guided ionization wave (conditions: He gas flow = 2slm; voltage frequency = 10kH; voltage amplitude = 5kV; voltage duty cycle = 1%.

<p>Optical fiber placed 5mm after the exit ceramic tube).</p

He-GIW induces a necrotic cell death.

<p><b>(a-c)</b> Presence of apoptosis was assayed by FACS analysis of Annexin V/PI dual staining. <b>(a)</b> An example of flow cytometry profile 6 h after treatment with He-GIW or with staurosporine, a positive control for apoptosis. X axis represents annexin-V-APC staining and Y axis PI staining. <b>(b)</b> Percentage of cells positive for Annexin V only (black), PI only (white) or both (grey) was measured 6 h and 24 h after treatment with He-GIW or staurosporine in standard cell culture conditions (c) or with 1% or 5% FCS. <b>(a-c)</b> are representative of 3 independent experiments. <b>(d)</b> Caspase 3 cleavage (red) was assayed by immunofluorescence 6h after He-GIW or staurosporine treatment. Cell nuclei are labeled with Hoechst and appear blue. <b>(e)</b> Caspase 3 cleavage was monitored by Western Blot for 2 h after He-GIW treatment. <b>(f)</b> Adherent cell percentage 6 h after He-GIW treatment in presence or absence of caspase inhibitor Z-VAD. Error bars represent S.E.M. of three independent experiments. p<0.05 (*), p<0.01 (**) and p<0.001 (***)</p

Expression of cytokines in SD rat brains infected by K173.

<p>Cerebral cytokine gene expression was assessed in ECM (n = 14), NoECM (n = 23) and Control brains (n = 7). Samples were collected on day ECM onset for ECM rats, after the parasitemia reached 30% for NoECM rats and for control rats at the same time as ECM and NoECM groups. Box plots represent medians and 25th and 75th percentiles. Among all the cytokines investigated only the gene expression of INFγ (p<0.001), IL10 (p<0.001), IL1β (p = 0.001), TNFα (p = 0.045), MIP1α (p<0.014) and MIP1β (p<0.001) were significantly different between infected and control rats. The symbol * indicates outliers.</p

Biochemical parameters of SD rats infected by K173.

<p>Globally significant differences between ECM, NoECM and control (CTRL) groups were observed for creatinine (p = 0.001) potassium (p<0.001) glycemia (p<0.001), calcium (p = 0.002), albumin (p<0.001) and total CO₂ (p<0.001). The ECM group was significantly different from the CTRL group for creatinine (p = 0.001), glucose (p = 0.037) and total CO₂ (p<0.001), from the NoECM_LP group only for potassium (p = 0.004) and from the NoECM_HP group for albumin (p = 0.001) and chloride (p = 0.0029). To suppress the influence of parameter units, the data are plotted on a standardized scale, e.g. mean = 0 and SD = 1.</p

Localization of parasites in cerebral vessels in ECM K173 infected SD rats.

<p>(<b>A</b>) Thrombosis. (<b>B</b>) Parasites were localized in peripheries of cerebral vessels in direct contact with the endothelium. <b>(C</b> and <b>D)</b> Extravascular hemorrhage with parasites (arrows) localized in peripheries. ECM brains were collected from rats without systemic lavage. Histological slices of ECM brains stained using hematoxylin-eosin.</p