Pharmacological targeting of C5a receptors during organ preservation improves kidney graft survival.

Cadaveric renal transplants suffer frequently from delayed graft function, which is associated with increased risk for long-term graft survival loss. One-third of kidney grafts that are stored in current organ preservation solutions experience delayed graft function, demonstrating the urgent need for improvement. Although ischaemic graft injury is complex in nature, complement activation is considered important to the process. Here we show that pharmacological targeting of the complement 5a receptor (C5aR) during cold ischaemia has a protective effect on early kidney graft survival, inflammation and apoptosis in a mouse model of syngeneic kidney transplantation. Graft survival of kidneys that were stored in University of Wisconsin solution in the presence of a C5aR antagonist increased from 29% to 57%. Increased graft survival was associated with less tubular damage and apoptosis, protection from sustained C5aR expression and decreased production of tumour necrosis factor-alpha and macrophage inflammatory protein-2. In a translational approach, we determined C5aR expression in paediatric living-related and cadaveric allografts. C5aR expression was significantly higher in all compartments of kidneys from cadaveric compared with kidneys from living-related donors. C5aR expression in cadaveric kidneys correlated positively with cold ischaemia time, renal dysfunction and the frequency of apoptotic tubular cells, suggesting a novel role for C5a in delayed graft function pathogenesis. Supplementing organ preservation solutions with C5aR inhibitors may improve early graft function following cadaveric kidney transplantation.

Precision of computer-aided volumetry of artificial small solid pulmonary nodules in ex vivo porcine lungs.

The purpose of this study was to investigate the precision of CT-based volumetric measurements of artificial small pulmonary nodules under ex vivo conditions. We implanted 322 artificial nodules in 23 inflated ex vivo porcine lungs in a dedicated chest phantom. The lungs were examined with a multislice spiral CT (20 mAs, collimation 16x0.75 mm, 1 mm slice thickness, 0.7 mm increment). A commercial volumetry software package (LungCARE VA70C-W; Siemens, Erlangen, Germany) was used for volume analysis in a semi-automatic and a manual corrected mode. After imaging, the lungs were dissected to harvest the nodules for gold standard determination. The volumes of 202 solitary, solid and well-defined lesions without contact with the pleura, greater bronchi or vessels were compared with the results of volumetry. A mean nodule diameter of 8.3 mm (+/-2.1 mm) was achieved. The mean relative deviation from the true lesion volume was -9.2% (+/-10.6%) for semi-automatic and -0.3% (+/-6.5%) for manual corrected volumetry. The subgroup of lesions from 5 mm to <10 mm in diameter showed a mean relative deviation of -8.7% (+/-10.9%) for semi-automatic volumetry and -0.3% (+/-6.9%) for manually corrected volumetry. We conclude that the presented software allowed for precise volumetry of artificial nodules in ex vivo lung tissue. This result is comparable to the findings of previous in vitro studies.

Thermoablation of colorectal liver metastases promotes proliferation of residual intrahepatic neoplastic cells.

BACKGROUND: Resection of liver neoplasms is believed to promote growth of residual intrahepatic neoplastic cells. As the effects of radiofrequency thermoablation (RFA) are still unknown, we aimed to compare the influence of RFA versus liver resection on residual intrahepatic neoplastic cells. METHODS: A primary metastasis was established by injection of syngenic CT-26 coloncarcinoma cells into the right liver lobe of Balb/C mice. Five days later, 3 x 10(5) GFP-transfected CT-26 tumor cells were injected intraportally, and the primary metastasis was treated by resection (group I, n = 7) or RFA (group II, n = 7). The effect of resection/RFA on the growth of single intrahepatic GFP neoplastic cells was evaluated by intravital microscopy 7 days later. RESULTS: Resection of a primary metastasis enhanced the proliferation of residual intrahepatic neoplastic cells, compared with the control group. RFA led to an increased survival of residual neoplastic cells (5% +/- 2% vs 1% +/- 1% single cells) and significantly promoted the proliferation of neoplastic cells, compared with resection (13% +/- 4% vs 2% +/- 2% micrometastases). CONCLUSIONS: RFA strongly promotes intrahepatic growth of residual neoplastic cells. On the basis of our findings, RFA should not be recommended as an alternative curative treatment to resection. Furthermore, if RFA is performed as palliative therapy, postinterventional chemotherapy may be advisable to overcome the stimulation of residual neoplastic cells by RFA.

[Clinical evaluation of a software for automated localization of lung nodules at follow-up CT examinations]. / Klinische Evaluation einer Software zur automatischen Lokalisation von Lungenrundherden in CT-Verlaufskontrollen.

PURPOSE: To evaluate a software algorithm for automated localization of pulmonary nodules at follow-up CT examinations of the chest and to determine factors influencing the rate of correctly matched nodules. MATERIALS AND METHODS: The "real-time automatic matching" (RAM) algorithm (Siemens LungCare trade mark software) was applied to 22 follow-up multirow-detector CT (MDCT) examinations in 11 patients (Siemens Somatom VolumeZoom, tube voltage 120 kVp; effective tube current 20 mAs (n = 18) or 100 mAs (n = 4); 4 x 1 mm detector configuration, 1.25 mm slice thickness; 0.8 mm reconstruction increment; standard lung kernel B50f) with a total of 190 lung nodules (mean diameter 6.7 +/- 3.5 mm, range 2 - 17 mm). The following nodule features were recorded: diameter, edge definition (well- or ill-defined), location (upper, middle or lower third; central or peripheral; right or left lung) and inspiration level (considered identical if the difference of diaphragm-apex distance between baseline and follow-up examination was < 5 %, otherwise it was considered different). A nodule was regarded as correctly localized if the marking box drawn by the software was visible on at least one slice together with the nodule and the center of the nodule was located inside the marking box. chi(2)-test was used to describe influence of nodule features on detection rate. Influence of nodule size was assessed using Mann-Whitney-U-Test. RESULTS: RAM correctly located 164 of 190 of all lung nodules (86.3 %). Detection rate did not depend on nodule location (left vs. right lung: p = 0.48; upper vs. middle vs. lower third: p = 0.96; peripheral vs. central: p = 0.47) or diameter (p = 0.30). Influence of inspiration level was highly significant (p < 0.001): nodules were detected in 100 % (146/146) for identical inspiration levels and in 40.9 % (18/44) for different inspiration levels. The observation of a significant better localization of ill-defined nodules (p = 0.028) corresponds to a statistical artifact due to the inhomogeneous distributions of this specific feature in our data. CONCLUSION: RAM is a valuable tool for follow-up of lung nodules at CT. Only very different inspiration levels influenced detection rate.

Multi-slice computed tomography as a screening tool for colon cancer, lung cancer and coronary artery disease.

Recent promising trials that use low-dose CT for the early detection of lung cancer have reinvigorated the interest in screening approaches. At the same time the development of fast image acquisition techniques, such as multislice CT, have sparked renewed interest in cardiac imaging within the radiological community. In addition to special cardiac capabilities, multislice CT has several other features such as high acquisition speed and low-dose requirements that may make this modality a universal radiological screening tool. Non-invasive disease detection is the radiologist's domain. In this paper we identify criteria for effective screening and apply these criteria to screening approaches with multislice CT when used for detection of three disease entities: colon cancer; lung cancer; and cardiovascular disease.

Activation, stimulation and uptake of bacterial ghosts in antigen presenting cells.

Bacterial ghosts have been shown to be an innovative system to prepare vaccines of various bacteria with all features of the intact bacterial cell envelopes, especially all antigenic epitopes, but also to target recombinant proteins inserted in the cell envelopes of the ghost preparations to specific antigen presenting cells. To investigate the activation of the antigen presenting cell by bacterial ghosts in more detail we studied the uptake of bacterial ghosts in dendritic porcine cells and RAW macrophages and the induction of inflammatory mediators or mediators directing the immune response in THP-1 human macrophage cell line. The synthesis of inflammatory macrophage mediators such as TNFalpha in the THP1 cell line was stimulated by a hundred-fold higher dose of ghosts from Vibrio cholerae than the corresponding LPS using ELISA-analysis. These results confirm in vivo experiments indicating no toxic effects of ghosts in rabbits even after intravenous administration in doses stimulating significant humoral responses. We were also able to see a significant activation of IL-12 indicated by the analysis of IL-12(p70) synthesis and IL-12(p40) mRNA accumulation. This interleukine is of special importance in the activation of cellular TH1 immune responses. A rapid uptake of bacterial ghosts in macrophages within 10-30 min could be confirmed by electron microscopy. As antigen presentation is especially effective in porcine dendritic cells (DC) and even a low capacity of antigen uptake is sufficient for an induction of immune responses we investigated uptake and activation of bacterial ghosts by DC. DC are known to be phagocytic in specific immature stages. We found a significant uptake of bacterial ghosts from Actinobacillus pleuropneumoniae (App) and V. cholerae conjugated with FITC (fluorescinisothiocyanate) within 2 h. These data suggest that bacterial ghosts effectively stimulate monocytes and macrophages for the induction of TH1 directed immune responses and dendritic cells treated with bacterial ghosts may serve as a promising vehicle for active immunization and immunotherapy in situ.

Corticotropin-releasing-hormone lacks analgesic properties: an experimental study in humans, using non-inflammatory pain.

The antinociceptive potency of corticotropin-releasing-hormone (CRH) has been established in several animal studies in which both central and peripheral sites of action were considered. However, there have not yet been any experimental trials, besides one attempt using clinical dental pain demonstrating the potential analgesic properties of CRH in humans. For this reason, we studied the effect of CRH on experimental heat pain sensitivity in 18 healthy men, using a double-blind, cross-over and placebo-controlled design. A dose of 100 microg (i.v.) was chosen because of its well-known neuroendocrine effects in humans. The pain parameters assessed were, visual analog scale (VAS) ratings for pain intensity and pain unpleasantness, pain thresholds and scores for discrimination ability. To differentiate between a direct analgesic effect of CRH and indirect effects via evoked hormonal responses in the hypothalamic-pituitary-adrenocortical (HPA) system (beta-endorphin, ACTH, cortisol), CRH was applied with and without a pre-treatment with dexamethasone. In neither of the two conditions was there any systematic change in our pain parameters. This failure to find any evidence suggesting an analgesic action of CRH or of the subsequent hormones of the HPA system was obtained despite the fact that CRH produced clear neuroendocrine responses such as increases in the plasma concentration of beta-endorphin and cortisol. It is unclear whether the lack of analgesic action of CRH is due to its non-existence in humans, due to the use of a pain model which does not assess minute changes in pain sensitivity and does not trigger substantial inflammatory responses, or due to an insufficient dose of CRH.

Dissociated central and peripheral release of vasopressin, but not oxytocin, in response to repeated swim stress: new insights into the secretory capacities of peptidergic neurons.

To investigate the effects of an ethologically-relevant stressor on central and peripheral release of arginine vasopressin and oxytocin, we forced adult male Wistar rats to swim for 10 min and simultaneously measured the release of the two peptides (i) within the hypothalamic supraoptic and paraventricular nuclei (by means of the microdialysis technique) and (ii) into the blood (by chronically-implanted jugular venous catheters). Forced swimming caused a significant rise in the release of arginine vasopressin and oxytocin within both the supraoptic nuclei (four-fold and three-fold, respectively) and the paraventricular nuclei (three-fold and four- to five-fold, respectively). Release patterns measured before, during and after repeated stress exposure on three consecutive days indicated that, at the level of the hypothalamus, the two neuropeptides are critically involved in the rats' stress response in a peptide-, locus- and stress-specific manner. Particularly, despite a general reduction of the recovery of the microdialysis probes over the time, the release of arginine vasopressin within the paraventricular nuclei and of oxytocin within the supraoptic nuclei tended to increase upon repeated stress exposure. Measurement of plasma peptide concentrations revealed that the central release of oxytocin was accompanied by a secretion of this peptide into the systemic circulation. In contrast, arginine vasopressin, assayed in the same plasma samples, failed to respond to the stressor. The latter finding is consistent with a dissociated release of the neuropeptide from different parts of a single neuron (soma/dendrites vs axon terminals). It provides evidence that under physiological conditions plasma hormone levels do not necessarily reflect the secretory activity of central components of the respective neuropeptidergic system.

Release of vasopressin from supraoptic neurons within the median eminence in vivo. A combined microdialysis and push-pull perfusion study in the rat.

The present study was designed to investigate whether or not arginine vasopressin (AVP) is released from magnocellular neurons within the median eminence (ME) in vivo. Urethane-anesthetized adult male Wistar rats were equipped with a microdialysis probe aimed at the supraoptic (SON) or paraventricular nucleus (PVN), a push-pull perfusion probe resting in the ME, and a blood microdialysis probe within the jugular vein. Dialysis of the SON (but not the PVN) with Ringer's solution containing 56 mmol l-1 K+ resulted in an increase in AVP release within the ME (to 492 +/- 192% of release during basal conditions, P < 0.05) and into blood (to 138 +/- 9%, P < 0.01) whereby the release probably occurred from axonal swellings and nerve terminals of supraoptic neurons which project through the internal zone of the ME to the posterior pituitary. The calculated amount of AVP released into the extracellular fluid of the ME was high enough (approximately 1 pg/microliter) to hypothesize that the neuropeptide could enter the portal blood capillaries in physiologically relevant concentrations. Taken together, the present study indicates that activation of magnocellular neurons is accompanied by release of AVP within the median eminence. We assume that AVP released in this way might mediate a communication between the hypothalamic-neuro-hypophysial system and the hypothalamic-pituitary-adrenal axis in response to selected stressful stimuli.