Lung Transplantation in Hungary From Cardiac Surgeons' Perspective.

Thoracic organ transplantation made a fresh start in Hungary with the first double lung transplant in December 2015. This major leap in Hungarian transplantation was preceded by almost 10 years of preparation, new infrastructure development, and structural changes not only at the organizational level but in human resources as well. In the following years, until recently, altogether 47 lung transplants were performed on 24 men and 23 women. The underlying pathologies were as follows: chronic obstructive pulmonary disease, 25; cystic fibrosis, 11; idiopathic pulmonary fibrosis, 7; as well as other diseases, including bronchiectasis, eosinophilic granuloma, lymphangioleiomyomatosis, and primary pulmonary hypertension in 4 cases. The youngest recipient was 13 and the oldest was 65 years old. Overall survival rates at 30 days and at 1 year were 96% and 82%, respectively. No patients were lost in the cystic fibrosis and other diseases group, whereas the 1-year survival rates of the chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis groups were 73% and 71%, respectively. The results show the robustness and viability of the program, although there is still opportunity for further improvement. In this short paper, we summarize the fields of possible further cooperation of thoracic and cardiac teams as well as future challenges facing the new Hungarian lung transplant program.

Antibody-Mediated Rejection in a Multiple Lung Transplant Patient: A Case Report.

Lung transplant is an effective way to treat many end-stage lung diseases. However, one of the main barriers of allograft organ transplant is still the immunologic rejection of transplanted tissue, which is a response of the HLA molecules. Rejection is a complex process involving both T-cell-mediated delayed-type hypersensitivity reactions and antibody-mediated hypersensitivity reactions to histocompatibility molecules on foreign grafts. We report the case of a 25-year-old female patient with cystic fibrosis who underwent 2 lung transplants because of her initial diagnosis and appearance of bronchiolitis obliterans syndrome after the first transplant. Only 13 months after the second transplant, despite the therapies applied, a new rejection occurred associated with high mean fluorescent intensity donor-specific antibody levels, which resulted later in the death of the patient. The present case draws attention to the importance of matching HLA molecules between donor and recipient in addition to immunosuppressive therapy.

Early Postoperative Problems After Lung Transplantation: First-Year Experiences in Light of the Newly Established National Hungarian Lung Transplantation Program.

Until December 2015, Hungarian patients' lung transplantations (LTXs) were done at the Medical University of Vienna. After several years of preparation, the National Hungarian Lung Transplantation Program was launched and the first milestone LTX was performed in Budapest on December 12, 2015. During the first 12 months, 18 lung transplantations took place in Hungary, including the first one. Data were retrospectively collected to analyze the early postoperative problems of the first 18 LTX patients of the newly launched Hungarian National Lung Transplantation Program. No patients with primary pulmonary hypertension and no children were transplanted during this period. We found that the postoperative problems of LTX differ from those of other huge thoracic surgeries both in a quantitative and a qualitative manner. We also reveal problems that are not present with other thoracic surgeries. The wide variety of problems during the early postoperative period after LTX can be managed by a highly organized and coordinated interdisciplinary teamwork.

Urgent Lung Transplantation in Severe Acute Respiratory Failure Based on Rapidly Progressive Interstitial Lung Disease: A Case Report.

Lung transplantation (LUTX) became a worldwide accepted standard therapy for certain well-defined chronic end-stage lung diseases. Until recently, patients on mechanical ventilation or extracorporeal life support techniques were hardly eligible for LUTX because of the inferior short-term results. However, a paradigm shift has occurred, and now these techniques represent bridging options to LUTX for listed patients. In the current practice, transplantation from the intensive care unit (ICU) is not extraordinary in patients on the waiting list. On the other hand, transplantation of an ICU patient who has previously been healthy without any chronic lung disease is still exceptional. Here we report a unique case of a 37-year-old woman without any relevant medical history who developed acute lung failure based on a cryptogenic organizing pneumonia. Her condition rapidly deteriorated and she required mechanical support, then she was bridged to transplantation on venovenous extracorporeal membrane oxygenation. She was listed for LUTX, and despite elevated panel-reactive antibody values, positive crossmatch LUTX was performed. Induction therapy, alemtuzumab, plasmapheresis, and intravenous immunoglobulin were administered. Her recovery was slow but finally she could be discharged from hospital in stable condition. After 2 months at home, she was readmitted to the hospital with respiratory failure from combined antibody-mediated rejection and infection. Before December 2015, the launch of Hungarian National Lung Transplantation Program, Hungarian patients were transplanted in Vienna. This case presents an exceptional example of national and international teamwork that aimed to save a young woman's life.

ESI-MS and MS/MS identification of the first ceramide analogues of fumonisin B1 mycotoxin from a Fusarium verticillioides culture following RP-HPLC separation.

Following the earlier detection of six new esterified fumonisin B1 (EFB1) isomers containing three acyl groups in a Fusarium verticillioides-inoculated rice culture, it was assumed that linoleic, palmitic or oleic acid esterifies one of the free OH groups on the fumonisin backbone. On the basis of the results of our recent investigations we now propose that these EFB1 isomers are actually 3-O- and 5-O-acyl derivatives of FB1 (3-O-linoleoyl-FB1, 5-O-linoleoyl-FB1, 3-O-palmitoyl-FB1, 5-O-palmitoyl-FB1, 3-O-oleoyl-FB1 and 5-O-oleoyl-FB1). A F. verticillioides strain was identified that produced not only O-acyl-FB1 isomers, but also low amounts of three N-acyl derivatives (N-linoleoyl-FB1, N-palmitoyl-FB1 and N-oleoyl-FB1), which eluted from the HPLC column after the six O-acyl compounds and in the same sequence as for the O-acyl compounds. The characteristic positive and negative ESI-MS/MS spectra obtained after solid-phase extraction of the culture extract facilitated identification of these N-acyl-FB1 derivatives. The biosynthesis of N-palmitoyl-FB1 by F. verticillioides was verified by spiking the culture extract with synthetic N-palmitoyl-FB1. This is the first report of the separation and mass spectrometric identification of the six O-acyl- and three N-acyl-FB1 derivatives extracted from a F. verticillioides culture.

Identification of the first fumonisin mycotoxins with three acyl groups by ESI-ITMS and ESI-TOFMS following RP-HPLC separation: palmitoyl, linoleoyl and oleoyl EFB1 fumonisin isomers from a solid culture of Fusarium verticillioides.

The aim of this study was to apply RP-HPLC/ESI-ITMS and RP-HPLC/ESI-TOFMS to investigate and characterise six new higher molecular weight fumonisins (three pairs of isomers) extracted from a Fusarium verticillioides-infected solid rice culture. The ITMS and ITMS² spectra clearly indicated the m/z values (960, 984 and 986) of the protonated molecules and the FB1 toxin-like structures of these compounds, respectively. Moreover, the data evaluation software of the TOFMS equipment unambiguously demonstrated the exact masses of the protonated molecules and the suggested empirical formulae (C50H89NO16, C52H89NO16 and C52H91NO16) of the new fumonisins, with mass accuracy in the range between 0.1 and -1.1 ppm. Subtraction of the empirical formula of FB1 toxin (C34H59NO15) from these formulae and correction for the mass of water split-off from the fumonisin molecule during ester formation resulted in the empirical formulae of the fumonisin backbone esterifying agents (fatty acids): C16H32O2 (palmitic acid, PA), C18H32O2 (linoleic acid, LA) and C18H34O2 (oleic acid, OA). We denoted the new compounds as esterified FB1 (EFB1) toxins, with the suggested names EFB1 PA, iso-EFB1 PA, EFB1 LA, iso-EFB1 LA, EFB1 OA and iso-EFB1 OA. The total amount of these new compounds comprised 0.1% of the FB1 concentration, which may be rated as significant when it is considered that these new components are significantly more apolar than earlier-described fumonisins, and their uptake into and toxicity elicited in the various tissues of living organisms may therefore also be significantly different from those of other fumonisins.

Fumonisin contamination and fumonisin producing black Aspergilli in dried vine fruits of different origin.

Aspergillus niger isolates are able to produce fumonisins in high quantities on agar media with a low water activity. Several agricultural products fit this criterion, including dried vine fruits, dates and figs. Data on the occurrence and role of this species in fumonisin contamination of agricultural products with high sugar content are needed to clarify the importance of A. niger in human health. The mycobiota and fumonisin contamination of various dried vine fruit samples collected from different countries were examined to clarify the role of black Aspergilli in fumonisin contamination of such products. All except two of the examined samples were contaminated with black Aspergilli. Species assignment of the isolates was carried out using sequence analysis of part of the calmodulin gene. The range of fumonisin isomers present in the raisins samples, and produced by A. niger isolates collected from dried vine fruits was also examined using reversed-phase high-performance liquid chromatography/electrospray ionization-ion trap mass spectrometry (RP-HPLC/ESI-ITMS). Among the 30 A. niger/A. awamori isolates identified, 20 were found to be able to produce fumonisins (average contamination: 5.16 mg/kg; range: 0.017-19.6 mg/kg). The average fumonisin content of the 7 dried vine fruit samples which were found to be contaminated by potential fumonisin producing black Aspergilli was 7.22 mg/kg (range: 4.55-35.49 mg/kg). The isolates produced several fumonisin isomers also present in the dried vine fruit samples, including fumonisins B(1-4), 3-epi-FB(3), 3-epi-FB(4), iso-FB(1), and two iso-FB(2,3) forms. Fumonisin B(1) was detected for the first time in A. niger cultures. Most of these isomers have previously only been identified in Fusarium species. Our data indicate that A. niger and A. awamori are responsible for fumonisin contamination of dried vine fruits worldwide. The observed levels of contamination are alarming and pose a new threat for food safety.

Prevention of Fusarium mycotoxin contamination by breeding and fungicide application to wheat.

The two main possibilities for decreasing toxin contamination were investigated in this paper. In the breeding section, we found that for resistance evaluation the ratio of Fusarium-damaged kernels is more important as this has a closer correlation with the deoxynivalenol (DON) content than the extent of Fusarium head blight (FHB). A high variability was found among lines from the non-Fusarium programme. A 50% decrease of DON contamination could be achieved by a simple and consequent resistance control. The tests with the variety candidates proved the same; therefore, the resistance screening for variety registration is an effective means to stop the spreading of highly susceptible genotypes. The resistance breeding programme showed an even larger DON decrease related to regular susceptible control varieties. Fungicide treatments were generally only weakly effective. However, it was shown that the testing methodology was poor, and with the optimal coverage spraying as much as 90% reduction of DON on small plot tests can be achieved. A farm-scale technology was also developed where the DON reduction as a mean for 3 years was higher than 70%, more than double the regular praxis data. To stabilize this efficacy, we need the most powerful fungicides, a nearly horizontal spraying of heads (like Turbo FloodJet nozzles) that receive the necessary coverage and so enough active ingredient to protect heads and the spraying technology should be kept rigorously. A combination of resistance and fungicide application can reduce DON contamination levels to below the European Union limit of 1.25 mg kg(-1) for levels which would otherwise be around 8-10 mg kg(-1). We think that this will solve most of the problems.

Influence of Wheat Cultivar, Species of Fusarium, and Isolate Aggressiveness on the Efficacy of Fungicides for Control of Fusarium Head Blight.

Attempts to control Fusarium head blight (FHB) with fungicides have been highly variable. Variability is caused by cultivar resistance, fungicide efficacy, fungicide coverage, timing, and pathogen aggressiveness. In this research, fungicides were tested on winter wheat cultivars with different levels of resistance to FHB using different isolates of Fusarium graminearum and F. culmorum to evaluate the role of host resistance and isolate aggressiveness on severity of FHB. Fungicides were applied to groups of wheat heads to provide full coverage. Incidence and severity of FHB was measured by the severity of head symptoms, percentage of Fusarium-damaged kernels (FDK), yield loss, and deoxynivalenol (DON) contamination. Development of FHB was affected by fungicides, cultivars, fungal isolates, and most of the two-way interactions of these variables. Among the fungicides tested, those containing tebuconazole tended to be more effective in reducing FHB. Reduction of disease in susceptible cultivars may not be adequate to produce marketable yields under conditions of high disease pressure. In most cases, if a fungicide reduced FHB visual symptoms, similar decreases were detected in yield loss, DON concentration, and FDK reaction. In 1998, an increase in DON contamination compared with the Fusarium check was observed with azoxystrobin and carbendazim on the more susceptible cultivar. This increase in DON with some fungicide requires additional research. Research to develop more resistant cultivars, better spraying technology, and more effective fungicides is also needed.

Influence of the polyphenolic tannic acid on the toxicity of the insecticide deltametihrin to fish. A comparative study examining both biochemical and cytopathological parameters.

Humics and pesticides are present in aquatic environment and the toxicological consequences of their chemical interaction is well studied. However, data concerning the mechanism of the biochemical action of humic-pesticide combinations are scarce, especially in vertebrates. Thus we have chosen to study the in vivo effects of the plant polyphenolic tannic acid and the pyrethroid insecticide deltamethrin [Decis] alone or in combination on hepatic xenobiotic-metabolizing enzyme activities and the associated redox-parameters in carp, as the complex assessment of these systems are regarded to serve as a relevant biomarker of environmental pollution. Stress effects and tissue damage were followed by determination of the plasma glucose level, the activities of plasma transaminases, and by electron microscopy. Tannic acid alone exerted weak prooxidant effect due to its marked antioxidant enzyme inhibitory activity. Deltamethrin, applied in a very low dose, induced oxyradical production in fish via activation of cytochrome P450 isozymes. This effect was promoted by the antioxidant enzyme inhibitory action of tannic acid, when the two chemicals were combined; however, the ultrastructural damage of the hepatocytes was reduced by the common cytoprotective capacity of the phenolic. Numerous humics are known to alter the toxicity of pesticides and their influence depends on their type and concentration. Therefore, our work taken together with other comparative studies may contribute to the assessment of the impact of humics in nature, especially in case of environmental pollution.

Cytokinin activities in cells of wheat inflorescence in depependence of its developmental stage.

We demonstrate that differences in the endogenous biologically active levels and patterns of cytokinins depend on the organ differentiation, i. e. immature inflorescence before meiosis and immature embryo 14 days after anthesis, respectively, in winter wheat of two genotypes: Grana and Almari. Two cell lines were subjected to analysis of cytokinin contents. Higher levels of endogenous cytokinin were found in cells derived from immature embryo (E) in comparison to cells obtained from immature inflorescence (I). The levels of zeatin ryboside ([9R]Z) and benzyladenosine ([9R]BAP) were predominant and isopentenyladenine (iP) was very low for the cells E of both genotypes. The cytokinin amounts in cells E and I of the zeatin group were smaller than those of other groups. We suggest that the concentration of cytokinins in tissue capable of generative development in winter wheat should be on a level that occurred in inflorescence cell lines, in comparison to immature embryo cell lines. These results indicate that the effect of the developmental stage of ears of wheat on the capability to flower in winter wheat plants regenerated in in vitro culture may be induced by the hormone level.

New results on the mass spectra of cinchona alkaloids

The electrospray ionization (ESI) mass spectra of 16 cinchona alkaloid compounds were studied for the first time. The electron ionization (EI) spectra of 22 cinchona alkaloids were also recorded, 14 of which had not been examined previously. In the case of EI the characteristic direction of the fragmentation is the scission of the C8-C9 bond. Under EI the cleavage of the C4'-C9 bond occurs only in the case of hydrogenated cinchona alkaloids, whereas the C9-O bond cleavage can be observed in the case of ester and ether derivatives. At a low capillary exit voltage (CapEx) in the ESI measurements there is no fragmentation, and only the [M + H](+) and in some cases the double protonated [M + 2H](2+) ions can be detected. On increasing the CapEx the characteristic primary direction is the cleavage of the C9-O bond, which was observed in the case of epialkaloids and esterified or etherified cinchona derivatives, respectively. Copyright 2000 John Wiley & Sons, Ltd.

Diterpenoids from Euphorbia peplus.

From a pro-inflammatory active extract of Euphorbia peplus, two new diterpene polyesters based on the pepluane and jatrophane skeletons were isolated, together with four known ingenane and jatrophane diterpenes. The structures were determined on the basis of extensive NMR studies. Ingenol 3-angelate, which was obtained for the first time from this plant, is an irritant toxin with high activity.

Identification of ethyl pyruvate and dihydrocinchonidine adducts by electrospray ionization mass spectrometry.

Several ethyl pyruvate and dihydrocinchonidine adducts, formed by non-covalent interactions with alkali cations, have been identified for the first time using electrospray ionization mass and tandem mass spectrometry. This type of adduct may have an important role in asymmetric reactions of pyruvates in the presence of cinchonas.

Analysis of deprenyl metabolites in the rat brain using HPLC-ES-MS.

Methylamphetamine and amphetamine, the two major metabolites of deprenyl in the rat brain were analyzed using HPLC method combined with electrospray-mass spectrometer. (-)-Deprenyl and (+)-deprenyl were orally administered to rats either in a single dose of 10 mg/kg, or three times a week for three weeks. The metabolites were determined in four different parts of the rat brain, such as in the frontal cortex, corpus striatum, hippocampus, and hypophysis. The ratio of methylamphetamine to amphetamine was also compared after (-)-deprenyl and (+)-deprenyl treatments.

Urinary excretion of deprenyl metabolites.

(+)-Deprenyl metabolites in rat's urine, such as nordeprenyl. methamphetamine amphetamine and p-hydroxy. methamphetamine were identified by HPLC-MS. After oral administration of 10 mg of pure (-)- and (+)-deprenyl to human volunteers, their urine was analyzed by gas chromatography. The concentration of methamphetamine was found to be overwhelming in the case of the (-)-isomer, while amphetamine and methamphetamine were excreted in equal amounts when (+)-deprenyl was administered. The metabolic processes of deprenyl resulted in metabolites possessing different lipophilicity, as it has been shown by planar displacement chromatography.

Trichothecene chemotypes ofFusarium graminearum isolated from corn in Hungary.

Twenty three strains ofFusarium graminearum isolated from corn were screened for their ability to produce type A and B trichothecenes and zearalenone (ZEA) on the solid substrate rice grains in the dark at 28 °C for 21 days. Toxin analyses were made with HPLC technique. Of 23 total isolates, 10 produced deoxynivalenol (DON), 4 produced DON and nivalenol (NIV), 1 produced DON and 15 acetyl-DON (15-ADON), 1 produced NIV and 4-acetyl-NIV (4-ANIV) and 1 produced NIV. Of 23 totalF. graminearum isolates, 20 produced ZEA. These results suggest that strains ofF. graminearum, prevailing in Hungarian corn growing regions, might belong to DON-and NIV-chemotypes. This is the first report demonstrating that DON-, DON-NIV-, DON-15-ADON-, NIV-4-ANIV and NIV-producingF. graminearum isolates are distributed in Hungary.