High-performance autotolerant bioelectrodes should be ideally suited to design implantable bioelectronic devices. Because of its high redox potential and ability to reduce oxygen directly to water, human ceruloplasmin, HCp, the only blue multicopper oxidase present in human plasma, appears to be the ultimate biocatalyst for oxygen biosensors and also biocathodes in biological power sources. In comparison to fungal and plant blue multicopper oxidases, e.g. Myrothecium verrucaria bilirubin oxidase and Rhus vernicifera laccase, respectively, the inflammatory response to HCp in human blood is significantly reduced. Partial purification of HCp allowed to preserve the native conformation of the enzyme and its biocatalytic activity. Therefore, electrochemical studies were carried out with the partially purified enzyme immobilised on nanostructured graphite electrodes at physiological pH and temperature. Amperometric investigations revealed low reductive current densities, i.e. about 1.65 µA cm−2 in oxygenated electrolyte and in the absence of any mediator, demonstrating nevertheless direct electron transfer based O2 bioelectroreduction by HCp for the first time. The reductive current density obtained in the mediated system was about 12 µA cm−2. Even though the inflammatory response of HCp is diminished in human blood, inadequate bioelectrocatalytic performance hinders its use as a cathodic bioelement in a biofuel cell.

Purpose Physical exercise is reported to affect the immune response in various ways. Thus, the levels of pro-inflammatory cytokines as well as the abundance of circulating leukocytes are changed. In this study, the occurence of circulating cell-free mitochondrial DNA (cfmtDNA) and nuclear DNA (nDNA) was investigated in connection with a single bout of strenuous physical exercise. Methods Healthy volunteers performed a controlled ergo-spirometry cycle test and venous blood samples were taken at different time-points to analyze the concentration of blood components before, during and after the test. The number of circulating leukocytes was measured, as well as secretion of the soluble urokinase activator receptor (suPAR). Results Cf-mtDNA significantly increased during exercise, compared to baseline values and after 30 and 90 min of rest. Circulating leukocytes increased during exercise, but returned to baseline levels afterwards. Surface expression of the urokinase plasminogen activating receptor (uPAR) on neutrophils decreased significantly during exercise. The concentration of suPAR tended to increase during exercise but only significantly after 90 min of rest. Conclusion Increased concentration of cf-mtDNA indicates that cell damage takes place during high intensity training. Hypoxia and tissue damage are likely causes of cf-mtDNA from muscle cells. The levels of cf-mtDNA remain high during the initial rest, due to the decreasing numbers of leukocytes normally clearing the plasma from cf-mtDNA. The increased levels of suPAR further emphasize that strenuous physical exercise causes a reaction similar to inflammation. Further studies are needed to detect the source of increased cf-mtDNA and the corresponding increase of suPAR liberation.

OBJECTIVE: Inflammation is associated with major depressive disorder (MDD) and suicidal behavior. According to the 'leaky gut hypothesis', increased intestinal permeability may contribute to this relationship via bacterial translocation across enterocytes. We measured plasma levels of gut permeability markers, in patients with a recent suicide attempt (rSA), MDD subjects with no history of a suicide attempt (nsMDD), and healthy controls (HC), and related these markers to symptom severity and inflammation. METHOD: We enrolled rSA (n = 54), nsMDD (n = 13), and HC (n = 17). Zonulin, intestinal fatty acid binding protein (I-FABP), soluble CD14, and interleukin-6 (IL-6) were quantified in plasma. Montgomery-Asberg Depression Rating Scale (MADRS) and Suicide Assessment Scale (SUAS) were used for symptom assessments. RESULTS: The rSA group displayed higher I-FABP and lower zonulin levels compared with both the nsMDD and the HC groups (all P < 0.001). IL-6 correlated positively with I-FABP (r = 0.24, P < 0.05) and negatively with zonulin (r = -0.25, P < 0.05). In all subjects, I-FABP levels correlated positively with MADRS (r = 0.25, P < 0.05) and SUAS scores (r = 0.38, P < 0.001), and the latter correlation was significant also in the nsMDD group (r = 0.60, P < 0.05). CONCLUSION: The 'leaky gut hypothesis' may improve our understanding of the link between inflammation and suicidal behavior. These findings should be considered preliminary until replicated in larger cohorts.

Thin macroporous poly(vinyl alcohol) (PVA) hydrogels were produced by cross-linking of PVA in a semi-frozen state with glutaraldehyde (GA) on glass slides or in the wells of microtiter plates. The 100-130 mu m-thick gels were mechanically transferable, squamous translucent films with a high porosity of 7.2 +/- 0.3 mL/g dry PVA i.e. similar to larger cylindrical PVA monoliths of the same composition. Additional treatment of the gels with 1% GA increased the aldehyde group content from 0.7 to 2.4 mu mol/mL as estimated using dinitrophenylhydrazine (DNPH) reagent. Translucency of the gels allowed registration of UV-visible spectra of the DNP' ;Wined films. The catalytic activity of trypsin covalently immobilized on thin gels in the microtiter plates was estimated with chromogenic substrate directly in the wells, and indicated that the amount of protein immobilized was at least 0.34 mg/mL gel. Human immunoglobulin G (IgG) immobilized on thin gels at 0.1-10 mg/mL starting concentrations could be detected in a concentration-dependent manner due to recognition by anti-human rabbit IgG conjugated with peroxidase and photometric registration of the enzymatic activity. The results indicate good permeability of the hydrogel pores for macromolecular biospecific reagents and suggest applications of thin reactive PVA hydrogels in photometric analytical techniques.

Background: Mitochondrial DNA copy number (mtDNA-cn), which represents the number of mitochondrial genomes per cell, can be quantified in peripheral blood mononuclear cells (PBMC) and is thought to reflect variations in mitochondrial biogenesis. Additionally, mtDNA may be released at low levels into the circulation from mitochondria under cellular stress, resulting in circulating cell-free mtDNA (ccf-mtDNA) detectable in plasma. The source or physiological significance of ccf-mtDNA in psychiatric illness is unknown but may reflect cell damage, cell death, or bioenergetic compromise. Methods: We enrolled suicide attempters (across diagnoses), non-suicidal subjects with Major Depressive Disorder (MDD), and healthy controls (all medication-free) in two independent cohorts (n=110 & n=74). MtDNA was quantified in cell-free plasma and in PBMCs. Results: Ccf-mtDNA was elevated in suicide attempters and in non-suicidal MDD subjects, compared to healthy controls. These group effects were very large (Cohen’s d ranging from 0.9 to 4.0, all p<0.00001). Ccf-mtDNA and cellular PBMC mtDNA-cn were not significantly correlated with each other (r=0.02, p=0.87), suggesting they reflect different processes. Ccf-mtDNA correlated with post-dexamethasone cortisol (r=0.5, p<0.001), suggesting that HPA-axis hyperactivity may be associated with cellular damage and release of ccf-mtDNA into the blood. Ccf-mtDNA also directly correlated with the antioxidant enzyme glutathione peroxidase (r=0.32, p=0.001), possibly reflecting a compensatory attempt to upregulate antioxidant defence mechanisms due to cellular stress. Conclusions: Ccf-mtDNA may represent a novel marker of cellular stress, which is increased in certain psychiatric conditions. These results call for replication in larger cohorts and in longitudinal studies.

Inflammation has been proposed to play a role in the generation of depressive symptoms. Previously, we demonstrated that patients with major depressive disorder (MDD) have increased plasma levels of the soluble form of the urokinase receptor (suPAR), a marker for low-grade inflammation. The aim of this study was to test the hypothesis that acute exercise would induce inflammatory response characterized by increased suPAR and elucidate whether patients with MDD display altered levels of suPAR in response to acute exercise. A total of 17 patients with MDD and 17 controls were subjected to an exercise challenge. Plasma suPAR (P-suPAR) was analyzed before, during, and after exercise. There was a significantly higher baseline P-suPAR in the patients with MDD, and the dynamic changes of P-suPAR during the exercise were significantly lower in the patients with MDD, compared with the controls. This study supports the hypothesis that an activation of systemic inflammatory processes, measured as elevated P-suPAR, is involved in the pathophysiology of depression. The study concludes that P-suPAR is influenced by acute exercise, most likely due to release from activated neutrophils.

Background: Preclinical data suggest that chronic stress may cause cellular damage and mitochondrial dysfunction, potentially leading to the release of mitochondrial DNA (mtDNA) into the bloodstream. Major Depressive Disorder has been associated with an increased amount of mtDNA in leukocytes from saliva samples and blood, but no previous studies have measured plasma levels of free-circulating mtDNA in a clinical psychiatric sample. Methods: In this study, free circulating mtDNA was quantified in plasma samples from 37 suicide attempters, who had undergone a dexamethasone suppression test (DST), and 37 healthy controls. We hypothesized that free circulating mtDNA would be elevated in the suicide attempters and associated with hypothalamic pituitary adrenal (HPA)-axis hyperactivity. Results: Suicide attempters had significantly higher plasma levels of free-circulating mtDNA compared to healthy controls at different time points (pre- and post-DST) (all p-values ,2.98E-12, Cohen’s d ranging from 2.55-4.01). Pre-DST plasma levels of mtDNA were positively correlated with postDST cortisol levels (rho50.49, p,0.003). Conclusions: Suicide attempters may have elevated plasma levels of free-circulating mtDNA, which are related to impaired HPA-axis negative feedback. This peripheral index is consistent with increased cellular or mitochondrial damage. The specific cells and tissues contributing to plasma levels of free-circulating mtDNA are not known, as is the specificity of this finding for suicide attempters. Future studies are needed in order to better understand the relevance of increased freecirculating mtDNA in relation to the pathophysiology underlying suicidal behavior and depression.

Here we report an enzymic fuel cell in a vein replica that generates sustained electricity, enough to power an e-​ink display, in an authentic human blood stream. We also detail a simple and safe approach for fuel cell evaluation under homeostatic conditions. Our results demonstrate proof-​of-​principle operation of a biocompatible and safe biodevice that could be implanted in superficial human veins, which we anticipate to be a starting point for more sophisticated investigations of personal sources of electricity.

Monolithic composites of Polyphepan (R) or Kraft lignin embedded in a poly(vinyl alcohol) (PVA) matrix were synthesized using cryogelation technique and studied as flow permeable adsorbents for bisphenol A and erythromycin removal from water. Adsorption isotherms of bisphenol A on pristine Polyphepan provided the equilibrium dissociation constant K-L = 2.6 x 10 (6) M and the maximal binding capacity Q(max) = 20 mu mol/g; for erythromycin K-L was in the 9.6 x 10 (6) M to 5.8 x 10 (5) M range, and Q(max) was between 55 mu mol/g and 94 mu mol/g. Embedment of lignins into PVA cryogels resulted in monoliths with adequate flow permeability and the composites essentially retained the binding capacity for both bisphenol A and erythromycin. Percolation of contaminated water through the monoliths resulted in 10-fold reduction of the pollutant concentrations within 12-70 column volumes of the effluent. Due to the higher loading of lignin, the Kraft lignin-PVA composite showed higher adsorption capacity for erythromycin than Polyphepan-PVA. Stability and reversible compression of the monoliths in the flow of water were studied. Limitations are associated with leakage of soluble lignin, strongly expressed in the case of Kraft lignin-containing composites. (C) 2016 Elsevier Ltd. All rights reserved.

Preclinical data suggest that chronic stress may cause cellular damage and mitochondrial dysfunction, potentially leading to the release of mitochondrial DNA (mtDNA) into the bloodstream. Major depressive disorder has been associated with an increased amount of mtDNA in leukocytes from saliva samples and blood; however, no previous studies have measured plasma levels of free-circulating mtDNA in a clinical psychiatric sample. In this study, free circulating mtDNA was quantified in plasma samples from 37 suicide attempters, who had undergone a dexamethasone suppression test (DST), and 37 healthy controls. We hypothesized that free circulating mtDNA would be elevated in the suicide attempters and would be associated with hypothalamic-pituitary-adrenal (HPA)-axis hyperactivity. Suicide attempters had significantly higher plasma levels of free-circulating mtDNA compared with healthy controls at different time points (pre- and post-DST; all P-values < 2.98E - 12, Cohen's d ranging from 2.55 to 4.01). Pre-DST plasma levels of mtDNA were positively correlated with post-DST cortisol levels (rho = 0.49, P < 0.003). Suicide attempters may have elevated plasma levels of free-circulating mtDNA, which are related to impaired HPA-axis negative feedback. This peripheral index is consistent with an increased cellular or mitochondrial damage. The specific cells and tissues contributing to plasma levels of free-circulating mtDNA are not known, as is the specificity of this finding for suicide attempters. Future studies are needed in order to better understand the relevance of increased free-circulating mtDNA in relation to the pathophysiology underlying suicidal behavior and depression.

The soluble form of the urokinase receptor, suPAR, has been suggested as a novel biomarker of low-grade inflammation. Activation of the immune system has been proposed to contribute to the development of depression and suicidal behavior. In order to identify depressed and suicidal individuals who could benefit from an anti-inflammatory treatment, a reliable biomarker of low-grade inflammation is vital. This study evaluates plasma suPAR levels as a biomarker of low-grade inflammation in patients with major depressive disorder and in patients who recently attempted suicide. The plasma suPAR and an established biomarker, C reactive protein (CRP) of suicide attempters (n = 54), depressed patients (n = 19) and healthy controls (n = 19) was analyzed with enzyme-linked immunosorbent assays. The biomarker attributes of sensitivity and sensibility were evaluated using ROC curve analysis. Both the depressed patients and suicide attempters had increased plasma suPAR. The levels of suPAR discriminated better between controls and suicide attempters than did CRP. In the future, plasma suPAR might be a superior prognosticator regarding outcome of treatment applying conventional antidepressants in conjunction with anti-inflammatory drugs.

AIMS: To investigate the association between experimental tooth clenching and the release of β-endorphin in patients with myofascial temporomandibular disorders (M-TMD) and healthy subjects. METHODS: Fifteen M-TMD patients and 15 healthy subjects were included and assigned an experimental tooth-clenching task. Venous blood was collected and pain intensity was noted on a visual analog scale. The masseter pressure pain threshold (PPT) was assessed 2 hours before the clenching task and immediately after. A mixed-model analysis of variance was used for statistical analyses. RESULTS: Significant main effects for time and group were observed for pain intensity and PPT, with significantly lower mean values of pain intensity (P < .001) and PPT (P < .01) after the clenching task compared with baseline. M-TMD patients had significantly higher pain intensity (P < .001) and significantly lower PPT (P < .05) than healthy subjects. No significant time or group effects were observed for the level of β-endorphin. Neither pain intensity nor PPT correlated significantly with β-endorphin levels. CONCLUSION: This experimental tooth-clenching task was not associated with significant alterations in β-endorphin levels over time, but with mechanical hyperalgesia and low to moderate levels of pain in healthy subjects and M-TMD patients, respectively. More research is required to understand the role of the β-endorphinergic system in the etiology of M-TMD

Dental caries is a localized, transmissible, pathological infection process that ends up in the destruction of hard dental tissue. Numerous reports have shown the close relationship between salivary levels of Streptococcus mutans and dental caries. As S. mutans, is considered to be the principle etiological agent of dental caries, the development of a quick and convenient method for detection and quantification of these bacteria from patient saliva samples would simplify diagnosis and treatment. The purpose of this study was to compare a new means of quantifying bacteria using FTATM Elute cards and Real-Time Polymerase Chain Reaction to a conventional culture-based assay using oral S. mutans as a model sample. A total of 60 different saliva samples were investigated. The results show a significant negative correlation between the two methods, with a correlation coefficient of −0.577 (Spearman’s Correlation) and p < 0.01. The method demonstrates a high sensitivity, specificity and reliable quantitative results, covering a large range of bacterial concentrations.

We investigated whether ingestion of probiotic bacteria could influence salivary IgA levels, specific anti-mutans streptococci IgA levels and specific antibodies towards the ingested probiotic bacterium. The study was a randomised, double-blind, placebo-controlled trial, where the test group (n = 11) received twice daily chewing of gum containing Lactobacillus reuteri (2 9 108 CFU per dose) and the control group (n = 12) received placebo. Resting saliva was collected before and after 12 weeks of treatment and 4 weeks after end of treatment. Total salivary IgA concentrations were measured by ELISA. Specific IgA reactivity was determined using a whole-cell ELISA. Results were expressed as % IgA per protein in saliva. The level of total IgA% per protein increased significantly between pretreatment levels (13.5%) and follow-up treatment levels (14.4%) within the test group only (P < 0.05). No changes were seen in the control group during the trial. The level of probiotic-reactive antibodies decreased significantly between pre- and post-treatment samples (from 12.2% to 9.0%, P < 0.05) in the test group. Similarly, the level of specific mutans streptococci antibodies decreased significantly between pre- and post-treatment samples (P < 0.05) in the test group only (for Streptococcus mutans from 20.1% to 15.0%; for Streptococcus sobrinus from 7.4% to 5.3%). Ingestion of probiotic bacteria might influence the adaptive immune response of the host.

Isothermal titration calorimetry (ITC) was used to study interactions between water vapour and the surface of thermally converted sodium bicarbonate (NaHCO3). The decarboxylation degree of the samples was varied from 3% to 35% and the humidity range was 54-100%. The obtained enthalpy values were all exothermic and showed a positive linear correlation with decarboxylation degrees for each humidity studied. The critical humidity, 75% (RHo), was determined as the inflection point on a plot of the mean-DeltaH kJ/mole Na2CO3 against RH. Humidities above the critical humidity lead to complete surface dissolution. The water uptake (m) was determined after each calorimetric experiment, complementing the enthalpy data. A mechanism of water vapour interaction with decarboxylated samples, including the formation of trona and Wegscheider's salt on the bicarbonate surface is proposed for humidities below RHo.

Abstract: It has been suggested that soluble urokinase plasminogen activator (suPAR) can be used as a marker of disease severity and risk of mortality in sepsis. The aim with the present study was to compare plasma levels of suPAR in patients with severe sepsis to control subjects and correlate it with the level of inflammatory activation, severity and mortality. Samples were collected from 27 sepsis patients at the intensive care unit (ICU), Lund, Sweden; 90-day mortalities were registered. The suPAR level was significantly elevated in sepsis patients compared to controls, but not significantly higher in nonsurvivors than survivors. Plasma levels of suPAR did correlate weakly with the SOFA score and myeloperoxidase (MPO) but not with CRP, PCT, IL-6 or IL-10 in patients with severe sepsis. The weak correlation between suPAR and other inflammatory markers might suggest that suPAR reflects general activation of the immune system rather than exerting inflammatory actions.

The soluble urokinase plasminogen activator receptor (suPAR) has been detected in blood, plasma, serum, urine, ovarian cystic fluid, and cerebrospinal fluid. Elevated suPAR levels in plasma have been associated with negative outcomes in various diseases, such as bacteremia, sepsis, SIRS, cardiovascular disease, cancer, and tuberculosis. The primary aim of this study was to investigate whether suPAR can be detected in saliva from healthy individuals and thus, if saliva suPAR can be related to plasma suPAR, CRP, BMI, or gender. Blood and unstimulated whole saliva was collected from 20 healthy individuals (10 female and 10 male, median age of 28 years; range 21–41). CRP and suPAR were measured with ELISA in saliva and serum/plasma. suPAR was detected in all saliva samples in the 5.2–28.1 ng/mL range, with a median value of 17.1 ng/mL. Saliva suPAR was significantly higher (P , 0.001) but not correlated to plasma suPAR in healthy young adults with normal plasma suPAR levels. suPAR and CRP levels were correlated in blood but not in saliva. No correlation was found between BMI, age, or gender and suPAR in saliva.

Avsikten med studien var att undersöka förekomsten av Lactobacillus reuteri i saliv ef- ter användandet av ett probiotiskt kosttillskott innehållande L. reuteri samt fastställa om L. reuteri har effekt på plackindex (PlI) samt på den supra- och subgingivala mikrofloran. Studien var dubbelblind, randomiserad, placebokontrollerad och löpte över 12 veckor. Totalt deltog 23 friska individer randomiserade i test- respektive kontrollgrupp. I försöket testades placebotuggummi mot tuggummi med L. reuteri (ATCC 55730 och ATCC PTA 5289). Tuggummi togs 2 gånger per dag, morgon och kväll efter tandborstning. Startda- gen och efter 12 veckor samlades vilosaliv samt plack supra- och subgingivalt. Dessutom bedömdes plackindex enligt Silness & Löe på 4 indextänder mesialt och distalt. Efter ytterligare 4 veckor gjordes en uppföljning med prov på saliv och bedömning av plack på indextänderna. Mikrobiologisk analys utfördes med hjälp av checkerboard DNA-DNA hybridiseringsteknik samt odling på selektiva medier för bestämning av laktobacill- populationen. Totala laktobacillhalten i saliv ökade signifikant i båda grupperna (p<0,05) med en signifikant ökning av L. reuteri i testgruppen (p=0,008). Efter 16 veckor påträffades ingen L. reuteri i saliv. Kontrollgruppen visade en signifikant ökning av PII efter 12 veckor (p=0,023), medan testgruppen inte visade någon förändring. I båda grupperna regist- rerades en signifikant ökning supra- och subgingivalt av de flesta bakteriearter, men det fanns ingen signifikant skillnad mellan grupperna för undersökta bakterier. Kvoten mellan ”onda/goda” supragingivala bakterier minskade i testgruppen men var inte sig- nifikant. Motsvarande kvot för subgingivala bakterier minskade signifikant i båda grup- perna. Slutsats: Intag av L. reuteri resulterade i förekomst av L. reuteri i saliv men bakterien måste tillföras kontinuerligt. Ingen signifikant förändring i den undersökta supra- och subgingivala mikrofloran kunde påvisas. Den signifikanta ökningen i PlI i kontrollgruppen utan motsvarande förändring i testgruppen kan tyda på en probiotisk effekt av L. reuteri i denna population

A promising approach in sepsis therapy is the use of peptides truncated from serum- and membrane-proteins with binding domains for LPS: antimicrobial peptides (AMPs). AMPs can be useful in combination with conventional antibiotics to increase killing and neutralize LPS. Although many AMPs show a high specifi city towards bacterial membranes, they can also exhibit toxicity, i.e. non-specifi c membrane lysis, of mammalian cells such as erythrocytes and therefore, unsuitable as systemic drugs. A way to overcome this problem may be an extracorporeal therapy with immobilized peptides. This study will compare neutralization of LPS using different AMPs in solution and when immobilized on to solid phases. The peptides ability to neutralize LPS-induced cytokine release in whole blood will also be tested. The peptides are truncated derivates from the known AMPs LL-37, SC4, BPI, S3Δ and CEME. Two different methods were used to immobilize peptides, biomolecular interaction analysis, and Pierce SulfoLink Coupling Gel. To investigate LPS binding in solution the LAL test was used. After whole blood incubation with LPS and AMPs ELISA was used to measure TNF α , IL-1 β and IL-6 production. The results suggest that immobilization of antimicrobial peptides does not inhibit their capacity to neutralize LPS, although there are differences between the peptides tested. Thus, peptides derived from LL-37 and CEME were more effi cient both in LPS binding and neutralizing LPS-induced cytokine production

SUMMARY BACKGROUND: Real-time perfusion (RTP) adenosine stress echocardiography (ASE) can be used to visually evaluate myocardial ischaemia. The RTP power modulation technique, provides images for off-line parametric perfusion quantification using Qontrast software. From replenishment curves, this generates parametric images of peak signal intensity (A), myocardial blood flow velocity (beta) and myocardial blood flow (Axbeta) at rest and stress. This may be a tool for objective myocardial ischaemia evaluation. We assessed myocardial ischaemia by RTP-ASE Qontrast((R))-generated images, using 99mTc-tetrofosmin single-photon emission computed tomography (SPECT) as reference. METHODS: Sixty-seven patients admitted to SPECT underwent RTP-ASE (SONOS 5500) during Sonovue infusion, before and throughout adenosine stress, also used for SPECT. Quantitative off-line analyses of myocardial perfusion by RTP-ASE Qontrast-generated A, beta and Axbeta images, at different time points during rest and stress, were blindly compared to SPECT. RESULTS: We analysed 201 coronary territories [corresponding to the left anterior descendent (LAD), left circumflex (LCx) and right coronary (RCA) arteries] from 67 patients. SPECT showed ischaemia in 18 patients. Receiver operator characteristics and kappa values showed that A, beta and Axbeta image interpretation significantly identified ischaemia in all territories (area under the curve 0.66-0.80, P = 0.001-0.05). Combined A, beta and Axbeta image interpretation gave the best results and the closest agreement was seen in the LAD territory: 89% accuracy; kappa 0.63; P<0.001. CONCLUSION: Myocardial isachemia can be evaluated in the LAD territory using RTP-ASE Qontrast-generated images, especially by combined A, beta and Axbeta image interpretation. However, the technique needs improvements regarding the LCx and RCA territories.

Interactions of bacterial and host products in activating the innate immune system is an important area to address. The role of lipoteichoic acid (LTA) in these interactions is particularly important because it is understudied in comparison to other factors. This study evaluated the effect of cationic peptides (CPs) on LTA-induced proinflammatory cytokine production in human whole blood and on purified leukocytes. Four different CPs of truncated derivatives from the known peptides LL37, BPI, and CP207 were used. Two of the CPs (IG33 and LL33), derivatives from LL37, potentiated S. aureus LTA induced TNFα, IL-6 and IL-1β production in whole blood. The release of TNFα was increased 30-fold after 16 hours incubation. Intact LL37 also increased LTA-induced TNFα and IL-1β in a time dependent manner. LTA in combination with either LL33 or IG23 demonstrated a synergistic enhanced TNFα and IL-1β secretion on isolated leukocytes but not on purified monocytes. When complexed with IG23 and LL33, the electrophoretic mobility of LTA was altered in a non-denaturating gel electrophoresis. LTA was disaggregated and migrated more rapidly, suggesting an amphiphilic effect of CPs on LTA. In conclusion, LTA synergizes with LL37 and its truncated derivatives and this may lead to proinflammatory cytokine production and cause problems in sepsis therapy.

Background: Real-time perfusion (RTP) contrast echocardiography can be used during adenosine stress echocardiography (ASE) to evaluate myocardial ischemia. We compared two different types of RTP power modulation techniques, angiomode (AM) and high-resolution grayscale (HR), with 99mTc-tetrofosmin single-photon emission computed tomography (SPECT) for the detection of myocardial ischemia. Methods: Patients with known or suspected coronary artery disease (CAD), admitted to SPECT, were prospectively invited to participate. Patients underwent RTP imaging (SONOS 5500) using AM and HR during Sonovue® infusion, before and throughout the adenosine stress, also used for SPECT. Analysis of myocardial perfusion and wall motion by RTP-ASE were done for AM and HR at different time points, blinded to one another and to SPECT. Each segment was attributed to one of the three main coronary vessel areas of interest. Results: In 50 patients, 150 coronary areas were analyzed by SPECT and RTP-ASE AM and HR. SPECT showed evidence of ischemia in 13 out of 50 patients. There was no significant difference between AM and HR in detecting ischemia (p = 0.08). The agreement for AM and HR, compared to SPECT, was 93% and 96%, with Kappa values of 0.67 and 0.75, respectively (p < 0.001). Conclusion: There was no significant difference between AM and HR in correctly detecting myocardial ischemia as judged by SPECT. This suggests that different types of RTP modalities give comparable data during RTP-ASE in patients with known or suspected CAD.

Aim: The aim of this stydy was to assess whether supplementation of Lactobacillus reuteri could have an impact on the oral microbiota. Material and Methods: Twent-three healthy people aged 29 to 63 years were included. A randomised double-blind placebo-controlled study was executed consisting of 12 persons in the test group which was given the study product twice a day for twelve weeks containing L. reuteri (an equal mix of ATCC 55730 and PTA 5289 at a total of 2x108 CFU/dose) and the control group of 11 persons having corresponding placebo. Pre and post of study period plaque index and oral health status (gingivitis, probing pocket depth, bleeding on probing) were measured together with sampling of supra-, subgingival plaque and saliva collection. Microbiological analysis was done by using checkerboard DNA-DNA hybridization technique and selective culturing for lactobacilli determination. Four weeks after the last intake of the product reassessments of plaque and saliva was performed. Results: No difference in general oral health could be observed between the groups after L. reuteri supplementation. Plaque index increased significantly in the control group after twelve weeks (p= 0.023). A significant increase in total Lactobacillus counts in saliva occurred in both groups (p<0.05). The probiotic intervention resulted in a significant increase of L. reuteri (p=0.008) corresponding to 13.8% of the total lactobacilli couont. A distribution ratio of 1:4 (ATCC 55730/ATCC PTA 5289) between the two installed L. reuteri strains in saliva was noticed. Termination of intervention resulted in a wash out of L. reuteri. A significant increase was found for most bacterial species in both groups and both in supra- and subgingival plaque during the test period. There was no significant difference detected for any of the bacterial species between the groups neither in plaque location. The ratio between "bad/good" supragingival bacteria decreased for the test group, however, not significant. The corresponding ratio for subgingival bacteria decreased significantly for both groups (p= 0.05)with no significant difference between the groups. Conclusions: The supplementation of L. reuteri did not affect general oral health. Presence of L. reuteri in saliva is only temporary and washed out after termination of intervention. Microbiologically no significant effect of the probiosis was observed. It can not be concluded whether L. reuteri was established in the plaque of the test group or not.

Aims: Real-time perfusion (RTP) adenosine stress echocardiography (ASE) can be used to visually evaluate myocardial ischaemia. The RTP power modulation technique angio-mode (AM), provides images for off-line perfusion quantification using Qontrast® software, generating values of peak signal intensity (A), myocardial blood flow velocity (β) and myocardial blood flow (Axβ). By comparing rest and stress values, their respective reserve values (A-r, β-r, Axβ-r) are generated. We evaluated myocardial ischaemia by RTP-ASE Qontrast® quantification, compared to visual perfusion evaluation with 99mTc-tetrofosmin singlephoton emission computed tomography (SPECT). Methods and Results: Patients admitted to SPECT underwent RTP-ASE (SONOS 5500) using AM during Sonovue® infusion, before and throughout adenosine stress, also used for SPECT. Visual myocardial perfusion and wall motion analysis, and Qontrast® quantification, were blindly compared to one another and to SPECT, at different time points off-line. We analyzed 201 coronary territories (left anterior descendent [LAD], left circumflex [LCx] and right coronary [RCA] artery territories) in 67 patients. SPECT showed ischaemia in 18 patients and 19 territories. Receiver operator characteristics and kappa values showed significant agreement with SPECT only for β-r and Axβ-r in all segments: area under the curve 0.678 and 0.665; P < 0.001 and < 0.01, respectively. The closest agreements were seen in the LAD territory: kappa 0.442 for both β-r and Axβ- r; P < 0.01. Visual evaluation of ischaemia showed good agreement with SPECT: accuracy 93%; kappa 0.67; P < 0.001; without non-interpretable territories. Conclusion: In this agreement study with SPECT, RTP-ASE Qontrast® quantification of myocardial ischaemia was less accurate and less feasible than visual evaluation and needs further development to be clinically useful.

Glycosaminoglycans (GAGs) are structurally heterogeneous negatively charged polysaccharides. Endothelial GAGs, also known as glycocalyx, are involved in capillary permeability. In rat venules stimulated with proinflammatory substances ex vivo, the GAG-containing proteoglycan, syndecan-1, is shed from the endothelium. We wanted to investigate if we could trace the same response during septic shock as reflected in the circulating GAG levels. Arterial plasma samples were collected from 18 consecutive septic shock patients admitted to our intensive care unit. Plasma GAGs were measured with an Alcian blue slot binding assay, and syndecan-1 levels were measured with enzymelinked immunosorbent assay. Effects of GAGs on the antibacterial activity of plasma were assessed by a radial diffusion assay. The median plasma GAG level was significantly higher in the septic shock patients than in matched controls (median [interquartile range], 2.7 2g/mL [1.9 Y 4.8 2g/mL] vs. 1.8 2g/mL [1.7 Y 2.0 2g/mL]). Furthermore, the GAG levels were significantly higher in nonsurvivors (4.6 2g/mL [3.1 Y 8.8 2g/mL], n = 8) than survivors (1.8 2g/mL [1.6 Y 2.6 2g/mL], n = 10). The syndecan-1 levels were also increased in the patients compared with controls (246 ng/mL [180 Y 496 ng/mL] vs. 26 ng/mL [23 Y 31 ng/mL]) and correlated to the cardiovascular Sequential Organ Failure Assessment (SOFA) score. The GAGs inhibited the endogenous antibacterial activity of plasma as well as isolated antimicrobial peptides. The concentrations required were in the same range as the GAG levels measured in the patients. These results show that the GAG levels are increased in septic shock patients, possibly reflecting peripheral endothelial cell damage. We also found that GAGs in relevant concentrations neutralize antimicrobial peptides in plasma.

The nature and role of human milk microbiota in the early colonization and protection of infants from infection is the subject of increasing research. This study investigated the occurrence of Lactobacillus reuteri in milk of nursing mothers living in rural or urban areas in different geografic locations. Breat milk samples were collected from 220 mothers, 6-32 days after delivery, and analysed for the presence of total lactobacilli and L. reuteri. In all, 50% of mothers from rural areas in Japan and Sweden were L. reuteri-positive, whereas mothers from urban areas in South Africa, Israel and Denmark had very low and non-detectable levels. Overall, 15% of mothers had detectable L. reuteri in their milk. There were no significant differences in the prevalence of total Lactobacillus or L. reuteri in women from rural and urban habitats in the participating countries.