Certification of the reference material IRMM-426 for radionuclides in dried bilberries.

A new reference material for the activity concentration of (137)Cs, (40)K and (90)Sr in a dried berry matrix was certified from a batch of bilberries collected in the vicinity of the Chernobyl nuclear power plant. Radionuclides in this material were metabolised by the plants, therefore no spiking had to be performed. The material was processed at IRMM and homogeneity and stability of the certified parameters were demonstrated. The certified property values for (137)Cs, (40)K and (90)Sr were determined in the course of a supplementary comparison, CCRI(II)-S8.

PACAP immunoreactivity in human malignant tumor samples and cardiac diseases.

Pituitary adenylate cyclase activating polypeptide (PACAP) is a pleiotropic and multifunctional neuropeptide having important roles in various physiological processes. Recent trends in PACAP research point to the clinical introduction of PACAP or its analogs/fragments possibly in the near future. Recently, we have shown the presence of PACAP in human plasma, milk, placenta, and follicular fluid samples. However, relatively few data are available on PACAP in human tissues from patients with different disorders. The aim of the present study was to determine, by radioimmunoassay, the tissue level of PACAP38-like immunoreactivity (LI) and PACAP27-LI in different primary non-small cell lung cancer, colon tumor samples, and in cardiac muscle samples from patients suffering from ischemic heart disease and valvular disorders. We also labeled the PAC1 receptors in human cardiac cells. All samples showed significantly higher PACAP38-LI compared with PACAP27-LI. We found significantly lower levels of PACAP38-LI and PACAP27-LI in tumoral and peripheral samples compared with normal healthy tissue in both lung and colon cancers. Further investigations are necessary to describe the exact function of PACAP in oncogenesis. We showed that PACAP38-LI and PACAP27-LI are significantly higher in ischemic heart diseases compared with valvular abnormalities, suggesting that PACAP might play a role in ischemic heart disorders.

Results of an international comparison for the determination of radionuclide activity in bilberry material.

Radioactivity found in wild food products has assumed greater importance when assessing the total exposure of the population. For this reason, IRMM has been developing a reference material for the activity concentration of three radionuclides in bilberry samples. In order to characterise this new material, a CCRI(II) supplementary comparison was organised. The difficulties encountered in this comparison are discussed, in particular the efficiency calibration for volume sources of gamma-ray emitters, and comparison reference values for (137)Cs and (40)K are calculated.

Complications after ultrasonic lung parenchyma biopsy: a strong note for caution.

BACKGROUND: This study aimed to determine the possible cause for an unacceptable frequency of postresectional pneumothorax in cases of ultrasonic scalpel use without a further reinforcing maneuver in lung biopsy during video-assisted thoracic surgery (VATS). METHODS: Data for a series of 16 consecutive VATS lung biopsy patients (group A) in which a disturbingly high number of minor and medium complications occurred were compared with data for a group of 20 patients previously subject to the same ultrasonic lung biopsy method (group B) without complication. RESULTS: The two groups were identical in terms of all significant factors considered in relation to ultrasonic scalpel biopsy. Six notable air leakage complications occurred among the 16 patients of group A. One patient needed redrainage while still in the hospital. Two other patients required readmission and redrainage. In 4 of the 16 cases, late pneumothorax was detected after a "silent" 48-h postoperative period prolonging their hospital stay. Altogether, three medium complications occurred in group A, as compared with none in group B. The drainage duration in group B was not significantly shorter than in group A . Multivariate analysis showed a significant difference in complications favoring group B (odds ratio, 1.88). CONCLUSIONS: A high postoperative air leakage rate was observed in a simple case series using an unsecured harmonic scalpel after a randomized trial of the same method in the same institute with a diametrically opposite outcome. The medium complication rate of 3 in 16 cases is unacceptable for a minor procedure such as lung biopsy. The two groups differed only in their thromboembolic prophylaxis protocol. Therefore, it is hypothesized that the recent introduction of low-molecular-weight heparin from day 1 may influence the complication rate. The authors' observation calls for caution in use of the harmonic scalpel on lung tissue without reinforcing maneuvers (i.e., stitches or clips). To avoid unnecessary complications, operative technique adjustment is recommended.

PKA-Bad-14-3-3 and Akt-Bad-14-3-3 signaling pathways are involved in the protective effects of PACAP against ischemia/reperfusion-induced cardiomyocyte apoptosis.

The neuropeptide PACAP (pituitary adenylate cyclase activating polypeptide) and its receptors are widely expressed in the nervous system and various other tissues. PACAP has well-known anti-apoptotic effects in neuronal cell lines. Recent data suggest that PACAP exerts anti-apoptotic effects also in non-neuronal cells. The peptide is present in the cardiovascular system, and has various distinct effects. The aim of the present study was to investigate whether PACAP is protective against in vitro ischemia/reperfusion-induced apoptosis in cardiomyocytes. Cultured cardiomyocytes were exposed to 60 min ischemia followed by 120 min reperfusion. The addition of PACAP1-38 significantly increased cell viability and decreased the ratio of apoptotic cells as measured by MTT test and flow cytometry. PACAP induced the phosphorylation of Akt and protein kinase A. In the present study we also examined the possible involvement of Akt- and protein kinase A-induced phosphorylation and thus inactivation of Bad, a pro-apoptotic member of the Bcl-2 family. It was found that ischemia significantly decreased the levels of phosphorylated Bad, which was counteracted by PACAP. Furthermore, PACAP increased the levels of Bcl-xL and 14-3-3 protein, both of which promote cell survival, and decreased the apoptosis executor caspase-3 cleavage. All effects of PACAP1-38 were inhibited by the PACAP antagonist PACAP6-38. In summary, our results show that PACAP has protective effects against ischemia/reperfusion-induced cardiomyocyte apoptosis and provides new insights into the signaling mechanisms involved in the PACAP-mediated anti-apoptotic effects.

Interpretation of scoring functions using 3D molecular fields. Mapping the diacyl-hydrazine-binding pocket of an insect ecdysone receptor.

A method is presented for the interpretation of receptor docking score values (rough measures of binding affinities) of ligands in terms of 3D molecular field interaction contributions. The FlexX and FlexX-Pharm methods were used to dock the structures of designed sets of ligands into the ligand-binding pocket of a selected receptor. In the next step the relationship was investigated between the FlexX and CScore scores and 3D molecular fields obtained for the docked conformations of the ligands, using the CoMFA (Comparative Molecular Field Analysis) and CoMSIA (Comparative Molecular Similarity Indices Analysis) methods. The approach yielded highly significant CoMFA and CoMSIA models demonstrating that a high portion of the variance in the docking score values of the ligands can be explained by steric, electrostatic, hydrophobic, and hydrogen bond donor and acceptor molecular field interaction contributions. The approach was exemplified by using the crystal structure of the ligand-binding domain of the ecdysone receptor (EcR) of the moth Heliotis virescens as well as virtual molecule libraries of analogues of known diacyl-hydrazine (DAH) type ecdysteroid agonists. By docking appropriately designed virtual compound libraries into the DAH binding pocket of EcR followed by CoMFA and CoMSIA of the docked conformations, hitherto unexplored regions of the receptor cavity could be mapped. By mapping the significant molecular field interaction contributions onto the model of the receptor-ligand complex, important receptor-ligand interactions could be highlighted that may help the design of novel highly scored receptor ligands. An advantage of the method is that no experimental biological activity data are required to exhaustively map the receptor-binding site.

Expression of CD97 and adhesion molecules on circulating leukocytes in patients undergoing coronary artery bypass surgery.

OBJECTIVE: Leukocyte activation is thought to be responsible for the adverse effects and postoperative complications following cardiopulmonary bypass (CPB). A novel cell surface molecule, CD97, is a sensitive marker of leukocyte and primary lymphocyte activation. The present study aimed to determine the activation of different leukocyte subsets by comparing the expression of CD97 and adhesion molecules (CD11, CD18) in patients receiving coronary surgery with or without CPB. METHODS: 30 patients were enrolled and scheduled for coronary bypass surgery under CPB (20 patients, group A) and with off-pump (OP) operation (10 patients, group B). Blood samples were taken before and during surgery, and over the following first week. RESULTS: Here, we report an early decrease in CD97 expression of granulocytes (PMN) and monocytes (MC) followed by an intensive increase reaching the maximum on postoperative days 2 and 3 in patients operated with CPB. The rate of active CD97-positive lymphocytes showed a marked, gradual increase until postoperative day 3 and remained elevated up to day 7 after CPB. OP surgery resulted in moderate alteration in the presence of CD97 on PMN, MC and lymphocytes. The expression of adhesion molecules was similar to CD97 in all leukocyte subsets. CONCLUSION: The findings about CD97 expression suggest considerable leukocyte activation following coronary bypass with CPB compared to OP surgery. The collected data show that the lymphocytes are highly activated and involved in leukocyte sequestration after CPB. Moreover, the importance of CD97 in CPB-related inflammatory response can be stated.

Dynamism of NF-kappaB and AP-1 activation in the signal transduction of ischaemic myocardial preconditioning.

Nuclear factor (NF)-kappaB and activation protein (AP)-1 transcription factors play an important role in the signal transduction of delayed ischaemic preconditioning (PC) leading to myocardial cytoprotection. Because the exact mechanism of the activation of these factors is still not clear, we aimed to monitor the time fluctuation of NF-kappaB and AP-1 induction in an in vivo animal model. Furthermore, we measured the induction rate of these factors using repeated cycles of PC. Following median thoracotomy, anaesthetized animals (24 New Zealand White rabbits) were subjected to ischaemic PC by occlusion of the left anterior descending coronary artery for 5 min. After 10 and 30 min, and 1, 2, 3 and 4 h of reperfusion, tissue samples were taken from the ischaemic myocardium, and the DNA binding activity of the transcription factors was measured with electrophoretic mobility shift assay. A further 12 animals were subjected to 2 x, 3 x or 4 x 5-min ischaemic PC, and after a 30-min or 1-hour reperfusion period, we investigated the possible modulation of NF-kappaB and AP-1 induction. Our results show significant, biphasically increased NF-kappaB activity with peak levels at 30 min and 3 h of reperfusion in preconditioned myocardium. AP-1 increased monophasically, with the peak level at 1 h of reperfusion. Repeated PC stimuli enhanced the activity of both transcription factors analyzed, but there was no significant correlation between the number of cycles and the rate of activation. Our results show that the activation of NF-kappaB and AP-1 have a specific time curve, and the induction of these factors is only slightly influenced by the number of PC cycles.

Mitigation of oxidative injury by classic and delayed ischemic preconditioning prior to small bowel autotransplantation.

Ischemic preconditioning (IPC) has been defined as short periods of ischemia with intermittent reperfusion. IPC induces two phases of protection. We sought to investigate the effects of classic and delayed preconditioning on oxidative stress markers prior to autotransplantation. Total orthotopic intestinal autotransplantation was performed on 18 mongrel dogs in three groups: group I (GI, nonpreconditioned), group II (GII, classic preconditioned), and group III (GIII, delayed preconditioned). In GI 3-hour cold preservation in University of Wisconsin solution was followed by 1 hour of reperfusion. In GII before this procedure the intestine was preconditioned by occlusion of the mesenteric artery with four cycles each of 5 minutes of ischemia and 10 minutes of reperfusion (IPC protocol). In GIII on day 1 the animals underwent the IPC protocol, and autotransplantation was performed on day 2. Oxidative stress parameters included malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD) measurements in tissue samples. Our results showed increased lipid peroxidation with decreased GSH level and SOD activity in GI (control: 254.38 +/- 18.32 IU/g; reperfused: 55.01 +/- 26.40 IU/g; P <.05). In GII MDA was slightly elevated, and the GSH concentration was increased markedly. Furthermore, better preservation of SOD activity was observed at the end of the reperfusion. Meanwhile, in GIII GSH was significantly increased, indicating the activation of the endogenous antioxidant protective system (control: 382.13 +/- 24.22 micromol/L per gram; reperfused: 515.25 +/- 26.36 micromol/L per gram; P <.05). Moreover, SOD surpassed the control activity. Our findings confirmed that both forms of preconditioning mitigate the severity of oxidative stress prior to preservation and autotransplantation. Delayed preconditioning is more effective to protect bowel tissue against oxidative injury.

Bradykinin and angiotensin-converting enzyme inhibition in cardioprotection.

OBJECTIVES: To show that angiotensin-converting enzyme (ACE) inhibition potentiates subthreshold ischemic preconditioning (IPC) via the elevation of bradykinin activity, leading to a fully delayed cardioprotective response. METHODS: On day 1 of the experiment, pigs were subjected to sham (group 1, controls) or IPC protocols. In groups 2 and 3, 4x5 min and 2x2 min of IPC, respectively, were elicited by occluding the left anterior descending coronary artery with percutaneous transluminal coronary angioplasty inflatable balloon catheter. Group 4 was subjected to the ACE inhibitor perindoprilate only. In group 5, the pigs were pretreated with perindoprilate (0.06 mg/kg) and then subjected to 2x2 min IPC. In group 6, intracoronary HOE 140 (a selective bradykinin B(2) receptor antagonist) was added before the perindoprilateaugmented subthreshold (2x2 min) PC stimulus. On the second day, all animals underwent 40 min left anterior descending coronary artery ligation and 3 h reperfusion, followed by infarct size analysis using triphenyl tetrazolium chloride staining. RESULTS: THE RATES OF INFARCT SIZE AND RISK ZONE WERE THE FOLLOWING IN THE EXPERIMENTAL GROUPS: group 1, 42.8%; group 2,19.5% (P<0.05); group 3, ischemia/reperfusion (I/R) 33.4%; group 4, I/R 18.4% (P<0.05); group 5, I/R 31.2%; and group 6, I/R 36.3%. A significant increase of nuclear factor kappa B activation in groups 2 and 4 was seen. CONCLUSIONS: Results confirm that ACE inhibitors do not give total pharmacological IPC, but they enhance the induction effect of small ischemic insults, which raises the ischemic tolerance of myocardium. It was determined that enhanced bradykinin activity leads to downstream nuclear factor kappa B activation in this model.

Effects of pretreatment with PACAP on the infarct size and functional outcome in rat permanent focal cerebral ischemia.

PACAP exerts neuroprotective effects under various neurotoxic conditions in vitro. In vivo, it reduces brain damage after global and transient focal ischemia. The present study investigated whether PACAP has neuroprotective effects when applied before the onset of permanent ischemia. Rats were given bolus injections of PACAP38 intracerebroventricularly, and then underwent permanent middle cerebral artery occlusion. The results show that 2 microg of PACAP significantly reduced the infarct size measured 12 and 24h after the onset of ischemia. No further reduction was obtained by a 7-day pretreatment. PACAP also ameliorated certain sensorimotor deficits. Our present study provides further evidence for the neuroprotective effects of PACAP, and implies that it might be a promising preventive therapeutic agent in ameliorating ischemic brain damage.

[Partial replacement of the trachea with jejunal autograft in the dog]. / Légcsópótlás vékonybél szabadlebennyel állatkísérletes modellben (Elókísérlet).

Extensive lesions of the cervical trachea can occur as a result of malformation, inflammation, tumor disease or trauma. If a short segment of the trachea is resected primary anastomosis may be possible. The problem of how to treat lesions longer than 50% of total tracheal length is still unresolved. The aim of our study was to clarify the possible use and limitations of small bowel interposition for tracheal replacement in veterinary model. We resected 6 cm of the cervical trachea of 5 adult mongrel dogs. Autolog jejunum free flap was used for replacement. The wall of the implant was supported with 5 polytetrafluoroethylene rings placed on the outer surface of the bowel perpendicular to its axis. We performed the surgery in two stages. Intraoperative tracheostomy was necessary in every case. The tracheal tube extended through the whole length of the implant. The survivals, the viability of the graft and complications were examined. The length of survival ranged between 18 h and 72 h. In one case obstruction of the tracheal tube occurred, in 3 cases the micro vessel anastomosis of the graft thrombosed, in one case both complications developed. In our experience use of the small bowel flap was complicated with fatal technical difficulties. The micro vessel anastomosis proved to be highly risky. Further investigations are needed to improve the results with this method.

[Monitoring of oxidative stress after experimental small bowel autotransplantation]. / Az oxidatív stressz monitorozása kísérletesen létrehozott vékonybél autotranszplantációt követóen.

The difficulty at transplanting the small bowel mainly is caused by the biology of the intestine. It is highly immunogenic, is one of the most sensitive tissues to ischemia-reperfusion injury. Our aims were to investigate changes of oxygen free radical mediated reactions after autotransplantation at different preservation times and perfusion fluids. Our results prove that this model is feasible to examine ischemia-reperfusion injury in the small intestine. Euro Collins (EC) is a suitable preserving solution for small bowel transplantation. There was no significant lipid peroxidation within the first 6 hours of graft preservation. However superoxide dismutase (SOD) activity was dramatically reduced during reperfusion in the tissues samples. Significant increase of reduced glutathione at the same time can be explained by compensatory mechanism to neutralize increased free radical production.

Traumatic myiasis of geese in Hungary.

Five geese flocks were surveyed to gather data on the prevalence and clinical manifestation of traumatic myiasis and the fly species involved. Myiasis was recorded in all the flocks and the total number of infested geese was 26 (ca. 0.1% of the total numbers). The first cases were observed at the end of May, the last ones in August. Most birds (16/26) were infested in August. Each affected goose had only one lesion, which was located more frequently on the wings (14/26) than on any other body. In seven geese, Wohlfahrtia magnifica (Diptera: Sarcophagidae) was the only myiasis-causing species. In these cases the detransformed mean number of larvae per wound was 18.1 (range 5-40). Lucilia sericata (Diptera: Calliphoridae) was found to be solely responsible for the lesions of 12 birds, with detransformed mean of 94.0 (range 2-893) larvae per goose. The larvae of this species appeared to be generally less invasive than those of W. magnifica, but in three cases they were also deeply embedded in the wounds. In seven geese larvae of both fly species developed together in and around the wounds. With the exception of one lesion, there were more larvae of W. magnifica (detransformed mean of 21.8 with a range of 1-55) than that of L. sericata (detransformed mean of 11.2 with a range of 2-61) in these mixed infections. Predisposing conditions for development of traumatic myiasis in geese included plucking of feathers, other injuries and bacterial infections (e.g. inflammation of the phallus).

Comparative three-dimensional quantitative structure-activity relationship study of safeners and herbicides.

The competitive antagonist hypothesis for safeners and herbicides was investigated by studying the 3D similarity between 28 safener and 20 herbicide molecules in their putative biologically active, low-energy conformations using comparative molecular field analysis (CoMFA). In addition, CoMFA provided information about the structural requirements for the interactions of safeners and herbicides with a proteinaceous component (SafBP) isolated from etiolated corn seedlings. Statistically significant CoMFA models have been developed for the united and separate safener and herbicide molecule sets using retrospective binding affinity data of the ligands measured at the SafBP receptor. The predictive power of the models was characterized by squared cross-validated correlation coefficients (q(2)) of 0.708, 0.564, and 0.4000 for the united safener plus herbicide set, the safener set, and the herbicide set, respectively. The CoMFA results support the competitive antagonist hypothesis between certain types of safeners and herbicides. The findings suggest that structural similarity between these two classes of agrochemicals is a useful guide in the design of new safeners.

Effect of the different factors on the iontophoretic delivery of calcium ions from bentonite.

The calcium content of mud patches used for therapy is very small. Several mineral clays originating from Hungary served as a base material for experiments in order to find a suitable drug for transdermal introduction of calcium ions into the body. The Ca++ transport through the pig skin has been investigated in vitro in diffusion cells applying iontophoresis. Studies of electrical and physicochemical factors acting on the permeation kinetics of in vitro experiments were performed. The utilization of direct current has intensified the Ca++ transport through the pig skin (129.78+/-26. 15 microgram Ca/cm2). On using pulsate currents the amount of the Ca++ penetrating through the skin was 5-10 times higher (283.18+/-16.89 microgram Ca/cm2, 388.71+/-19.90 microgram Ca/cm2) than that of the passive transport (36.22+/-14.20 microgram Ca/cm2). The amount of Ca++ cumulated in the receptor compartment was directly proportional to the amount of bentonite (a natural mineral clay with a large cation exchange capacity) in the donor compartment and to the concentration of Ca++ in the lattice of the applied mineral clay. Therefore, the experiments were carried out on a bentonite previously enriched in Ca++ in its lattice (50 mg Ca/g bentonite). The results of the in vitro studies could open a new field of application in the therapy of osteoporosis or in the use of mineral substances.