Beta Release Research Articles

The chondroprotective effect of selective COX-2 inhibition in osteoarthritis: ex vivo evaluation of human cartilage tissue after in vivo treatment

Recent in vitro studies showed that celecoxib, a selective cyclooxygenase (COX)-2 inhibitor, protects human osteoarthritic cartilage tissue from degeneration. The objective was to substantiate these beneficial effects in an in vivo (clinical) study with celecoxib treatment of patients with severe knee osteoarthritis (OA) and subsequent evaluation of cartilage tissue ex vivo. Patients with knee OA were treated 4 weeks prior to total knee replacement surgery with either celecoxib 200mg b.d., indomethacin 50mg b.d., or received no treatment. During surgery cartilage and synovium were collected and analyzed in detail ex vivo. When compared to non-treated patients, patients treated with celecoxib showed significant beneficial effects on proteoglycan synthesis, -release, and -content, confirming the in vitro data. In the indomethacin group, no significant differences were found compared to the control group. On the contrary, a tendency towards a lower content and lower synthesis rate was found. In the treated groups prostaglandin-E(2) levels were lower than in the control group, indicating COX-2 inhibition. Ex vivo release of interleukin-1 beta (IL-1 beta) and tumour necrosis factor-alpha by synovial tissue was decreased by treatment with celecoxib, whereas in the indomethacin group only IL-1 beta release was decreased. Using this novel approach we were able to demonstrate an in vivo generated chondrobeneficial effect of celecoxib in patients with end stage knee OA.

A Free Radical Scavenger, Edaravone, Prevents Ischemia-Reperfusion Injury in Liver Grafts From Non–Heart-Beating Donors

Background Due to the increase in liver transplantation, the donor shortage has become a serious problem, requiring marginal, non–heart-beating donors (NHBDs). The aims of this study were to evaluate the cytoprotective effect of edaravone, a free radical scavenger, on warm ischemia-reperfusion (I/R) injury of liver grafts from NHBDs. Methods Rat livers were harvested from heart-beating donors (HB group) or from NHBDs undergoing cardiac arrest for 30 minutes led by thoracotomy (NHB group), and reperfused for 60 minutes with Krebs-Henseleit bicarbonate buffer after cold preservation for 6 hours. In another group (ED group), warm ischemic livers from NHBDs were reperfused with buffer containing edaravone (1 mg/L) after cold preservation. Results In the ED group, portal flow volume, bile production, and energy charge were significantly ameliorated. Lipid peroxidation, elevation of hepatic enzymes, and release of tumor necrosis factor-alpha and interleukin-1 beta were significantly alleviated, compared with the NHB group. Conclusions These results suggested that edaravone has suppressive effects on warm I/R injury in liver grafts from NHBDs.

α-Synuclein enhances secretion and toxicity of amyloid beta peptides in PC12 cells

α-Synuclein is the fundamental component of Lewy bodies which occur in the brain of 60% of sporadic and familial Alzheimer’s disease patients. Moreover, a proteolytic fragment of α-synuclein, the so-called non-amyloid component of Alzheimer’s disease amyloid, was found to be an integral part of Alzheimer’s dementia related plaques. However, the role of α-synuclein in pathomechanism of Alzheimer’s disease remains elusive. In particular, the relationship between α-synuclein and amyloid beta is unknown. In the present study we showed the involvement of α-synuclein in amyloid beta secretion and in the mechanism of amyloid beta evoked mitochondria dysfunction and cell death. Rat pheochromocytoma PC12 cells transfected with amyloid beta precursor protein bearing Swedish double mutation (APPsw) and control PC12 cells transfected with empty vector were used in this study. α-Synuclein (10 μM) was found to increase by twofold amyloid beta secretion from control and APPsw PC12 cells. Moreover, α-synuclein decreased the viability of PC12 cells by about 50% and potentiated amyloid beta toxicity leading to mitochondrial dysfunction and caspase-dependent programmed cell death. Inhibitor of caspase-3 (Z-DEVD-FMK, 100 μM), and a mitochondrial permeability transition pore blocker, cyclosporine A (2 μM) protected PC12 cells against α-synuclein or amyloid beta evoked cell death. In contrast Z-DEVD-FMK and cyclosporine A were ineffective in APPsw cells containing elevated amount of amyloid beta treated with α-synuclein. It was found that the inhibition of neuronal and inducible nitric oxide synthase reversed the toxic effect of α-synuclein in control but not in APPsw cells. Our results indicate that α-synuclein enhances the release and toxicity of amyloid beta leading to nitric oxide mediated irreversible mitochondria dysfunction and caspase-dependent programmed cell death.

The RAVE/VERTIGO vertex reconstruction toolkit and framework

A detector-independent toolkit for vertex reconstruction (RAVE1) is being developed, along with a standalone framework (VERTIGO2) for testing, analyzing and debugging. The core algorithms represent state-of-the-art for geometric vertex finding and fitting by both linear (Kalman filter) and robust estimation methods. Main design goals are ease of use, flexibility for embedding into existing software frameworks, extensibility, and openness. The implementation is based on modern object-oriented techniques, is coded in C++ with interfaces for Java and Python, and follows an open-source approach. A beta release is available.

Fas / FasL system, IL‐1β expression and apoptosis in chronic HBV and HCV liver disease

The Fas / Fas-ligand (FasL) system is an important death signal pathway in the liver. An enhanced local inflammatory response prompted by FasL expression, which contributes to neutrophil recruitment and interleukin-1 beta (IL-1beta) release, seems to be crucial to chronic liver damage, persistence of viral infections, and probably initiation and / or promotion of HCC. In order to evaluate the expression of Fas, FasL, and IL-1beta in different stages of human liver disease and to determine whether hepatitis B virus (HBV) and hepatitis C virus (HCV) infections modulate their expression, also in relation to apoptosis, we examined 87 liver samples obtained from patients with: chronic hepatitis (CH) (n.42), cirrhosis (n.9) and hepatocellular carcinoma (HCC) (n.16) and corresponding peritumoural tissues (n.16); histologically-normal liver (n.4) as controls. Fas, FasL and IL-1beta mRNA were quantified using reverse transcriptase-polymerase chain reaction. The apoptotic index was evaluated by TUNEL analysis. Our data showed a progressive Fas / FasL increase from CH to cirrhosis followed by a decline from the latter to HCC. In histological sections apoptosis was detected in HCC. A significant difference emerged between HCV and HBV-related disease for IL-1beta expression only in CH. A significant positive correlation between IL-1beta and FasL in HCV-related disease (P = 0.014) and an inverse correlation between IL-1beta and Fas in HBV-related disease (P = 0.021) were observed. The different pattern of IL-1beta, Fas and FasL expression found in HCV- and HBV-mediated liver disease, points to a different modulation of immune response B and C virus induced, while the decline in Fas / FasL expression in HCC may be related to defence mechanisms adopted by HCC cells against the immune system.

Enhanced gastric IL-18 mRNA expression in Helicobacter pylori-infected children is associated with macrophage infiltration, IL-8, and IL-1β mRNA expression

To investigate IL-18 mRNA expression in the gastric mucosa in Helicobacter pylori-infected children and its association with macrophage infiltration, IL-8, and IL-1 beta mRNA expression. From 39 children, blood samples were taken for IL-1 beta gene polymorphism analysis and antral biopsies were obtained for histology (including macrophage immunostaining), culture and semiquantitative analysis of IL-18, IL-8, IL-1 beta, and CD14 mRNA expression by reverse transcription-PCR (RT-PCR). RT-PCR was used for H. pylori ureA and cagA mRNA detection in gastric tissue. H. pylori-infected patients had significantly higher IL-18, IL-8, and IL-1 beta transcript levels and macrophage numbers in the antral mucosa than H. pylori-negative children. IL-1 beta-511/31 gene polymorphism had no impact on gastric IL-1 beta mRNA levels. IL-18 mRNA expression correlated with mRNA expression of IL-8 and IL-1 beta, and transcript levels of all three cytokines were associated with macrophage infiltration and CD14 mRNA expression in the gastric tissue. Significant correlation was also observed between macrophage numbers and histological parameters of gastritis. These results suggest that interleukin(IL)-18 and macrophages may have an important function in gastric inflammatory response to H. pylori infection in children. IL-18, and possibly CD14 receptor signalling pathway, may be involved in macrophage activation and subsequent IL-8 and IL-1 beta release.

Interleukin-1β Secretion From Cord Blood Mononuclear Cells In Vitro Involves P2X7 Receptor Activation

The purinergic receptor P2X(7) is activated by extracellular adenosine 5'-triphosphate (ATP) and promotes the efficient release of interleukin-1 beta. The authors examine protein and molecular expression of the P2X(7) receptor and its ability to stimulate interleukin-1 beta release in cord blood mononuclear cells (CBMCs) from placentae of term nonlaboring and laboring women. They show both mRNA and protein (78 kDa) expression for the P2X( 7) receptor in CBMCs of parturient and nonparturient women. Costimulation of CBMCs in vitro with bacterial endotoxin and ATP resulted in significantly (P < .05) enhanced interleukin-1 beta release in laboring (54.17 +/- 24.78 pg/mL(-1); n = 8) compared with nonlaboring (13.60 +/- 3.20 pg/mL(-1); n = 7) deliveries. Release of interleukin-1 beta in both groups was blocked by preincubation with oxidized ATP, a P2X(7) receptor antagonist. The authors provide evidence for a novel inflammatory pathway in the release of interleukin-1 beta, which may be linked to the onset of labor.

Agonist potency at P2X7 receptors is modulated by structurally diverse lipids

The P2X(7) receptor exhibits a high degree of plasticity with agonist potency increasing after prolonged receptor activation. In this study we investigated the ability of lipids to modulate agonist potency at P2X(7) receptors. A variety of lipids, including lysophosphatidylcholine, sphingosylphosphorylcholine and hexadecylphosphorylcholine were studied for their effect on P2X(7) receptor-stimulated ethidium bromide accumulation in cells expressing human recombinant P2X(7) receptors and on P2X(7) receptor-stimulated interleukin-1 beta (IL1 beta) release from THP-1 cells. The effects of the lipids were also assessed in radioligand binding studies on human P2X(7) receptors. At concentrations (3-30 microM) below the threshold to cause cell lysis, the lipids increased agonist potency and/or maximal effects at P2X(7) receptors in both ethidium accumulation and IL1 beta release studies. There was little structure activity relationship (SAR) for this effect and sub-lytic concentrations of Triton X-100 partially mimicked the effects of the lipids. The lipids caused cell lysis and increased intracellular calcium at higher concentrations (30-100 microM) which complicated interpretation of their effects in functional studies. However, the lipids (3-100 microM) also increased agonist potency 30-100 fold in radioligand binding studies. This study demonstrates that a diverse range of lipids increase agonist potency at the P2X(7) receptor in functional and binding studies. The broad SAR, including the effect of Triton X-100, suggests this may reflect changes in membrane properties rather than a direct effect on the P2X(7) receptor. Since many of the lipids studied accumulate in disease states they may enhance P2X(7) receptor function under pathophysiological conditions.

Hormonal regulation of bone growth and remodelling.

Many systemic and local hormones influence bone growth and remodelling. These include calcium regulating hormones, systemic growth regulators and local growth factors. Parathyroid hormone (PHT) is a potent stimulator of osteoclastic bone resorption and a direct inhibitor of osteoblastic collagen synthesis. However, intermittent low-dose PTH administration can increase bone formation in vivo. PTH may act indirectly via local factors. It has been shown to increase prostaglandin E2 (PGE2) and transforming growth factor beta (TGF-beta) release from bone. Both PGE2 and TGF-beta have complex effects on bone metabolism and are likely to be physiological regulators of bone remodelling. Oestradiol has been shown to inhibit bone resorption in vivo but not in vitro. While there is evidence for oestrogen receptors in cultured bone cells, the effect could still be indirect. Oestradiol can inhibit bone PGE2 release in an in vivo-in vitro model in the rat. Glucocorticoids are potent inhibitors of bone formation and inhibit PGE2 and interleukin 1 production both in vivo and in vitro. While many regulatory factors affect prostaglandin production in bone, the complex effects of PGE2 on bone metabolism make it difficult to predict the ultimate response. The major effects of PGE2 are stimulation of bone formation and resorption and an increase in bone turnover. However, opposite effects can occur at certain times and concentrations. Interactions among these factors could explain some physiological, pathological, and therapeutic responses in skeletal tissue.

P2 purinoceptors in the immune system.

Immune cells express plasma membrane receptors for extracellular nucleotides. Both G protein-linked metabotropic and channel-forming ionotropic receptors have been described, although no P2 receptor subtype has been cloned from the immune system thus far. Metabotropic receptors have been described in human B but not T lymphocytes; they have not been found in mouse B and T cells. Ionotropic receptors seem to be ubiquitously expressed in the immune system; however, their functional properties, if not their pharmacology, appear to be different in different immune cells. Human T normal and B leukaemic lymphocytes, human macrophages, mouse B and T lymphocytes, mouse microglial and macrophage cells, and rat mast cells express ionotropic receptors that recognize ATP4- as the preferred ligand, are activated by 3'-O-(4-benzoyl)benzoyl ATP and inhibited by oxidized ATP. The pharmacological profile of ionotropic receptors expressed by different immune cells is similar, but their permeability properties may be different: the pore formed by receptors expressed by macrophages, microglial cells and mast cells is typically permeable to charged molecules of molecular mass up to 900 Da; on the contrary, that expressed by lymphocytes has a molecular cut-off of 200-300 Da. The ionotrpic receptor of immune cells is modulated by inflammatory cytokines (e.g. interleukin [IL]-2 and gamma-interferon) and is also modulated during monocyte to macrophage differentiation. Transient stimulation of the ionotropic receptor of macrophages and microglial cells elicits IL-1 beta release. Sustained activation leads to cell death, either by necrosis or apoptosis, depending on the given cell type.

RAVE—a Detector-independent vertex reconstruction toolkit

A detector-independent toolkit for vertex reconstruction (RAVE 1 1 RAVE = “reconstruction (of vertices) in abstract versatile environments”, ) is being developed, along with a standalone framework (VERTIGO 2 2 VERTIGO = “vertex reconstruction toolkit and interface to generic objects”. ) for testing, analyzing and debugging. The core algorithms represent state of the art for geometric vertex finding and fitting by both linear (Kalman filter) and robust estimation methods. Main design goals are ease of use, flexibility for embedding into existing software frameworks, extensibility, and openness. The implementation is based on modern object-oriented techniques, is coded in C ++ with interfaces for Java and Python, and follows an open-source approach. A beta release is available.

Cryptanalysis of an elliptic curve cryptosystem for wireless sensor networks

We present a brute-force attack on an elliptic curve cryptosystem implemented on UC Berkley's TinyOS operating system for Wireless Sensor Networks (WSNs). The attack exploits the short period of the Pseudorandom Number Generator (PRNG) used by the cryptosystem to generate private keys. In order to define failure in the event a brute-force attack takes too long to execute, we create a metric that relates the duty cycle of the mote to the compromise rate and the period of the key generation algorithm. Experimental results show that roughly 50% of the mote's address space leads to a private key compromise in 25 min on average. Furthermore, approximately 32% of the mote address space leads to a compromise in 17 min on average, 11% in 6 min and the remaining 7% in 2 min or less. We examine two alternatives to the PRNG our own design modified from a published algorithm and the new PRNG distributed with the beta release of TinyOS 2.0. Our design executes 12.47 times faster than the other alternative and requires 50 CPU cycles more than the original PRNG. In addition, our design is 6.3 times smaller than the other alternative and requires 106 additional bytes of memory. The period of our PRNG is uniform for all mote addresses and requires 6.6 years on average for a key compromise with the attack presented in this paper.

Functional Role of the Chemokine Binding Duffy Antigen Receptor Complex (DARC) in Human Inflammation In Vivo.

Background: The Duffy receptor is a promiscuous receptor for chemokines that binds selected members of both the C-X-C and C-C families with high affinity. Duffy antigen may influence plasma levels of proinflammatory cytokines by acting as a “chemokine sink”. Additionally, Duffy knockout mice experienced an exaggerated response to endotoxin in comparison to wild-type mice. The current trial was designed to elucidate the functional role of the Duffy blood group antigen in human inflammation. We hypothesized that “Duffy negative“ volunteers might show an increased inflammatory response to endotoxin (LPS) infusion in terms of cytokine response. The human endotoxemia model is a well established model of systemic inflammation, where a well defined, self-limited inflammatory stimulus permits the elucidation of key players involved in the inflammatory response. We therefore used this model to investigate the functional role of the Duffy Antigen Receptor Complex (DARC) in the inflammatory response after endotoxin challenge.Methods: Thirty-two healthy male volunteers received an intravenous infusion of 2ng/kg endotoxin, 16 Caucasians (Duffy antigen positive on erythrocytes) and 16 subjects of African descent (“Duffy negatives”). Cytokines, chemokines, as well as their receptors were quantified by ELISA, RT-PCR and FACS.Results: Plasma levels of TNF, IL-6, IL-8 and IL-8 mRNA in whole blood increased to a similar extent in both groups after LPS infusion. In contrast, peak MCP-1 levels at 3 hours were roughly 2-fold higher in Duffy positive subjects 16ng/mL as compared to Duffy negative subjects (7ng/mL p<0.001). Similarly, GRO-alpha levels were 2.5-fold higher in Duffy positive subjects at 2 hours after LPS infusion (210pg/mL vs 85pg/mL; p= 0.0001), whereas baseline values showed no difference between both groups. There was no difference in the LPS induced release of MIP-1 beta (which was used as a negative control). Baseline levels of MCP-1 and GRO-alpha in erythrocyte lysates were significantly higher in Duffy positive individuals, whereas IL-8 and MIP-1beta levels were comparable between both groups. Values of MCP-1 and GRO-α, both known to bind to the Duffy antigen, increased extensively in erythrocyte lysates of Duffy positive subjects over time (P=<0.001 vs. time). In contrast, MCP-1 only moderately changed over time in Duffy negative subjects (P<0.03 vs. time), whereas GRO-alpha remained comparable to baseline at all time points (P=0.2 vs. time). As a consequence, MCP-1 and GRO-alpha were several hundred-fold higher in Duffy positive subjects at all time points and reached peak levels at 2–4 hours. Additionally, IL-8, which also binds to erythrocytes via the Duffy antigen, was roughly 7-fold higher in erythrocyte lysates of Duffy positive subjects 2 hours after LPS infusion. As expected, MIP-1beta remained similar at all time points and was comparable between both groups. The expected increase in CD11b, and decrease in CCR-2 and CXCR-1, CXCR-2 occurred without differences between groups.Conclusion: This study characterized for the first time the in vivo function of the Duffy Antigen in humans. The blood group antigen has a profound effect on erythrocyte-bound chemokines and free chemokines in a model of human endotoxemia.

Does Off-pump Coronary Revascularization Reduce the Release of the Cerebral Markers, S-100β and NSE?

The aims of this study were to (1) compare the release of S-100 beta and NSE in off-pump coronary artery bypass grafting (CABG) versus on-pump surgery; (2) investigate whether the S-100 beta and NSE serum concentrations correlate with cardiopulmonary bypass (CPB) duration. Between October 2002 and May 2004, 42 patients undergoing first time CABG surgery were enrolled in the study. The exclusion criteria were: LVEF<35%, age>70 years, previous myocardial infarction, REDO surgery, the presence of valvular heart disease and/or cerebrovascular disease, abnormal preoperative carotid vessels angiography, coronary artery disease involving the distal circumflex artery, renal dysfunction, coagulopathy. The patients were randomly assigned either to undergo on-pump CABG surgery [group I, n=24 patients] or off-pump CABG [group II, n=18 patients]. Blood was not re-transfused from the cardiotomy suction. All patients presenting haemolysis were excluded from the study. The preoperative S-100beta was 0.13+/-0.08 (microg/l) and NSE 7+/-1.5 (microg/l) in group I and 0.12+/-0.1 (microg/l) and 6.9+/-2.7 (microg/l), respectively in group II. Six hours after the surgery, S-100beta in patients of group I reached a maximum level of 1.38+/-0.4 (microg/l) and NSE of 17.7+/-6.5 (microg/l) compared to 0.5+/-0.11 (microg/l) [S-100B] and NSE 8.6+/-4.2 (microg/l) in group II (p=0.001). Three (12%) patients in group I and none (0%) in group II suffered postoperative delirium, p=0.247. No strokes occurred linear regression analysis revealed a strong correlation between cardiopulmonary bypass duration and S-100beta and NSE peak levels, p<0.0021 (r(2)=0.36) and p<0.0001 (r=0.81), respectively. Coronary artery bypass surgery with CPB causes a significantly greater increase in NSE and S-100beta serum levels than off-pump surgery and correlates with CPB duration.

Nonorthogonal decomposition of binary matrices for bounded-error data compression and analysis

This article presents the design and implementation of a software tool, PROXIMUS, for error-bounded approximation of high-dimensional binary attributed datasets based on nonorthogonal decomposition of binary matrices. This tool can be used for analyzing data arising in a variety of domains ranging from commercial to scientific applications. Using a combination of innovative algorithms, novel data structures, and efficient implementation, PROXIMUS demonstrates excellent accuracy, performance, and scalability to large datasets. We experimentally demonstrate these on diverse applications in association rule mining and DNA microarray analysis. In limited beta release, PROXIMUS currently has over 300 installations in over 10 countries.

Drug Targets in Stress-Related Disorders

Nervous and immune systems mutually cooperate via release of mediators of both neurological and immunological derivation. Adrenocorticotropin hormone (ACTH) is a product of the hypothalamus-pituitary adrenal axis (HPAA) which stimulates secretion of corticosteroids from adrenals. In turn, corticosteroids modulate the immune response in virtue of their anti-inflammatory activity. On the other hand, catecholamines, products of the sympathetic nervous system (SNS), regulate immune function by acting on specific beta-adrenergic receptors. Conversely, cytokines released by monocytes/macrophages and lymphocytes, upon antigenic stimulation, are able to cross the blood-brain-barrier, thus modulating nervous functions (e.g., thermoregulation, sleep, and appetite). However, cytokines are locally produced in the brain, especially in the hypothalamus, thus contributing to the development of anorexic, pyrogenic, somnogenic and behavioural effects. Besides pathogens and/or their products, the so-called stressors are able to activate both HPAA and SNS, thus influencing immune responses. In this respect, many studies conducted in medical students taking exams have evidenced an array of stress-induced immune alterations. Phobic disorders and migraine without aura (MWA) represent examples of stress-related disorders in which phagocytic immune deficits, endotoxemia and exaggerated levels of proinflammatory cytokines [Tumor Necrosis Factor-alpha (TNF- alpha), and interleukin- 1 beta] have been detected. Quite interestingly, administration of a thymic hormone could ameliorate clinical symptoms in phobic patients. In MWA patients, a beta-blocker, propranolol, could mitigate migraine, whose cessation coincided with a drop of TNF-alpha serum concentration. In phobic disorders and in MWA, benzodiazepines are very often administered and, in this respect, some of them, such as diazepam, inhibit immune functions, while others, e.g., alprazolam, enhance immune responses. Alprazolam could improve clinical symptoms in MWA patients. Chronic Fatigue Syndrome (CFS) is a disorder whose etiology and pathogenesis are still unknown. In this syndrome both abnormalities of nervous and immune systems have been reported. Despite many immune parameters evaluated in CFS no specific biomarkers of disease have been found. Our own data are in agreement with current literature in that we found decreased levels of serum (IFN)-gamma in these patients, thus indicating a predominance of T helper (h)1 response in CFS. Also leptin, a hormone which regulates food intake, fluctuates within normal ranges in CFS individuals. Quite interestingly, in depressed patients, used as controls, leptinaemia was more elevated than in CFS. Finally, in a series of recent therapeutic trials several immunomodulating agents have been used, such as staphypan Berna, lactic acid bacteria, kuibitang and intravenous immunoglobulin. In conclusion, it seems that major drug targets in stress-related disorders are immune cells in terms of inhibition of proinflammatory cytokines and modulation of Th responses. In particular, according to recent evidences, antidepressants seem to exert beneficial effects in experimental autoimmune neuritis in rats by decreasing IFN- beta release or augmenting NK activity in depressed patients.

In vitro IL‐1β release from gingival fibroblasts in response to pure metals, dental alloys and ceramic

Little information is available on the immunological basis for side-effects of dental materials. The objective of this study is to evaluate effects of pure metals, dental alloys and ceramic on cell viability and interleukin-1 beta (IL-1beta) release in three-dimensional human gingival fibroblast cultures as an indicator of their biological performance in gingival tissues. The gingival fibroblast cultures were exposed to test specimens fabricated from nickel, iron, molybdenum, copper, indium, gold, Ni-Cr-Mo alloy (Remanium CS), Au-Pt-In alloy (Pontostar) and a dental ceramic (In-ceram). Cell viability was determined by the MTT method 24 and 48 h after exposure. Assays for IL-1beta were carried out by ELISA. Statistical analysis was performed applying the non-parametric Mann-Whitney pairwise test. Dental ceramic and gold did not influence cell viability after 24 and 48 h. Cell viability was determined after 24 and 48 h to nickel (79-77%), iron (92-90%), molybdenum (86-83%), copper (48-36%), indium (90-90%), Remanium CS (83-80%), Pontostar (94-91%) compared with control cultures. Dental ceramic, Pontostar and gold had no significant influence on IL-1beta secretion. The highest amounts of IL-1beta (10-fold) levels were determined in cell cultures exposed to copper. Indium, molybdenum and iron induced twofold IL-1beta levels compared with untreated control cultures. These results support that some metals may alter immune responses and thereby contribute to a variety of dental pathological conditions and three-dimensional cell culture models for gingival fibroblasts appear to be suitable for in vitro studies.

Anteil des frühen systolischen Flussanstiegs am antegrad fließenden Gesamtvolumen bei Phasenkontrast-Flussmessungen in Atemanhaltetechnik

Purpose: To evaluate the contribution of early systole for the assessment of antegrade aortic flow volume by breath-hold velocity-encoded magnetic resonance (MR) flow measurements. Materials and Methods: Expiratory breath-hold fast low-angle shot (FLASH) phase-contrast flow measurements (temporal resolution tRes 61 msec, shared phases) perpendicular to the proximal ascending aorta and short axis true fast imaging with steady-state precession (TrueFISP) cine MR ventriculometry (tRes 34.5 msec) were performed in ten healthy male volunteers on a 1.5 T MR system (Sonata, Siemens Medical Solutions). Antegrade aortic flow volume (AFV) and left ventricular stroke volume (LV-SV) were evaluated using Argus Ventricular Function and Argus Flow Software, version MR 2002B (Siemens Medical Solutions). A beta release of Argus Flow MR 2004A allowed interpolation of the flow up-slope during early systole to the preceding R-wave trigger. The respective intraindividual median differences between the AFV of each flow evaluation and LV-SV as well as between both AFV measurements were calculated and compared using the sign test for paired samples. Results: Non-interpolated AFV significantly deviated from LV-SV (p=0.006), underestimating the latter by 13.1 mL (13%). Interpolating aortic flow during early systole significantly increased AFV by 10.8 mL (13 %) compared to the flow evaluation which did not take early systole into account (p = 0.006). AFV with interpolation of early systolic flow agreed well with LV-SV (median difference -3.0mL or -3%, respectively), and no significant difference between these measurements was found (p = 1.0). Conclusion: Flow during early systole contributes substantially to total forward flow volume in the ascending aorta. Interpolation of the early systolic up-slope is therefore recommended for the evaluation of breath-hold phase-contrast flow measurements.

Anteil des frühen systolischen Flussanstiegs am antegrad fließenden Gesamtvolumen bei Phasenkontrast-Flussmessungen in Atemanhaltetechnik

To evaluate the contribution of early systole for the assessment of antegrade aortic flow volume by breath-hold velocity-encoded magnetic resonance (MR) flow measurements. Expiratory breath-hold fast low-angle shot (FLASH) phase-contrast flow measurements (temporal resolution tRes 61 msec, shared phases) perpendicular to the proximal ascending aorta and short axis true fast imaging with steady-state precession (TrueFISP) cine MR ventriculometry (tRes 34.5 msec) were performed in ten healthy male volunteers on a 1.5 T MR system (Sonata, Siemens Medical Solutions). Antegrade aortic flow volume (AFV) and left ventricular stroke volume (LV-SV) were evaluated using Argus Ventricular Function and Argus Flow Software, version MR 2002B (Siemens Medical Solutions). A beta release of Argus Flow MR 2004A allowed interpolation of the flow up-slope during early systole to the preceding R-wave trigger. The respective intraindividual median differences between the AFV of each flow evaluation and LV-SV as well as between both AFV measurements were calculated and compared using the sign test for paired samples. Non-interpolated AFV significantly deviated from LV-SV (p = 0.006), underestimating the latter by 13.1 mL (13 %). Interpolating aortic flow during early systole significantly increased AFV by 10.8 mL (13 %) compared to the flow evaluation which did not take early systole into account (p = 0.006). AFV with interpolation of early systolic flow agreed well with LV-SV (median difference - 3.0 mL or - 3 %, respectively), and no significant difference between these measurements was found (p = 1.0). Flow during early systole contributes substantially to total forward flow volume in the ascending aorta. Interpolation of the early systolic up-slope is therefore recommended for the evaluation of breath-hold phase-contrast flow measurements.

Differential growth factor retention by platelet-rich plasma composites

This study evaluates the temporal sequence and growth factor release from platelet-rich plasma (PRP) combined with different bone substitutes (BS), to identify an optimal substrate for extended growth factor retention. PRP was clotted with bovine thrombin or thrombin receptor activator peptide-6 (TRAP). In addition, PRP was clotted using Allogro (Ceramed, Lakewood, CO), BioGlass (Mo-Sci, Rolla, MN), or BioOss (Osteohealth, Shirley, NY). The effects of media exchange and BS on platelet-derived growth factor (PDGF) and transforming growth factor-beta (TGF beta) release were quantified via enzyme-linked immunosorbent assay. At day 1, the thrombin group released 36% more PDGF than the TRAP group and 80% more than the BS groups. At 7 days, PDGF release was the greatest for the TRAP group. PDGF release was minimal for all groups at day 14, with BS groups retaining 60% more PDGF than thrombin clots. Similarly, the thrombin group released the greatest amount of TGF beta (81.4% of the total), whereas TRAP and BS groups released significantly less TGF beta at day 1. Compared with thrombin, TRAP retained 39.2% more TGF beta, whereas BS groups retained even greater levels (Allogro, 54.3%; BioOss, 45.8%; BioGlass, 67.0%). No significant difference in TGF beta release was observed among the substitutes after day 1. The BS groups continued to retain TGF beta after 14 days, whereas all TGF beta in the thrombin clots was depleted. PRP preparation with thrombin results in a large, immediate release of growth factors that could be lost into the interstitium in vivo. TRAP-BS may prove more efficacious than thrombin in sustaining growth factor levels critical for the cascade of events leading to bone formation.