Zinkserumresponse beim Pferd nach oraler Verabreichung von unterschiedlichen Zinkverbindungen

Summary In the here presented study, the zinc serum response was investigated after the application of 4 different zinc compounds with varying dosages. To be able to consider species differences, dogs and cats were also supplemented with the same compounds and the zinc serum values measured. A total of 4 ponies, 12 dogs and 11 cats were alternately fed with zinc oxide, zinc sulfate, zinc lactate and B-Traxim using a single dose of 10 mg zinc/kg KM and the zinc serum levels measured over the next 24 hours. For the ponies, the effect of a single dose of 20 mg zinc/kg KM (except zinc lactate) was also measured. In the third approach, the ponies were supplemented 2,5 mg zinc/kg KM per day for a total of 14 days. The results were as follows: • The zinc serum level increased significantly after the application of zinc sulfate and B-traxim in relation to the dosage (Tab. 1). Zinc lactate also showed signs of a potential increase but due to the reduced tolerance was only tested in the lower dosages. In contrast, zinc oxide did not show a significant zinc serum response regardless of the applied amount. Tab. 1: Mean values of the maxima with standard deviation (SD) and minimum / maximum zinc serum values for the ponies (Mean values, not indicated with the same letter differ significantly) 10 mg Zn / kg KM 20 mg Zn / kg KM Compound Mean Maxima SD Range Mean Maxima SD Range Control 860 200b 660-1102 854 209b 649-1146 Zinc oxide 1100 340ab 776-1564 1136 463b 842-1785 Zinc sulfate 2080 800a 1030-2805 4134 1323a 2880-5888 B-Traxim 2120 670a 1397-2885 4895 1497a 3136-6206 Zinc lactate 1810 630ab 1171-2638 • No significant increase of the zinc serum levels could be found after the daily application of 2,5 mg zinc/kg KM over the 2 weeks. All levels were within the range of the ones presented in current literature. • For the dogs and cats, no significant zinc serum response could be determined after the supplementation of 10 mg zinc/kg KM, regardless of the zinc compound. For the cats, it could be speculated that the zinc serum response showed a certain tendency to higher serum levels after the application of zinc sulfate, B-traxim and zinc lactate. For these investigations, it can be concluded that zinc sulfate and B-traxim are available in the horse and are thus suited for the use in studies on zinc supplementation in regards to hoof quality. But one needs to consider, that from the results presented in this study, it can not be excluded that zinc oxide or zinc lactate are also available in the horse under certain circumstances.Zusammenfassung In der hier vorliegenden Arbeit wurde der Zinkserumresponse nach der Verabreichung von 4 unterschiedlichen Zinkpräparaten und gleichzeitig der Einfluss von variierender Dosierung untersucht. Um tierartliche Unterschiede mit berücksichtigen zu können, wurden Fleischfresser mit den gleichen Präparaten supplementiert und auch deren Zinkserumwerte bestimmt. Es wurden insgesamt 4 Ponys, 12 Hunde und 11 Katzen jeweils abwechselnd mit Zinkoxid, Zinksulfat, Zinklaktat und B-Traxim in einer Dosierung von 10 mg Zink/kg KM einmalig gefüttert und deren Zinkserumwerte über einen Zeitraum von 24 Std. untersucht. Bei den Ponys wurde ausserdem der Effekt auf den Zinkserumspiegel über 24 Std. nach einer einmaligen Supplementierung von 20 mg Zink/kg KM (Aussnahme Zinklaktat) untersucht. Als 3. Versuchsansatz wurde bei den Ponys nach einer täglichen Zugabe von 2,5 mg Zink/kg KM über 14 Tage der Zinkserumspiegel dokumentiert. Die Ergebnisse stellen sich wie folgt dar: • Beim Pony stieg der Serumzinkspiegel nach Verabreichung von Zinksulfat und B-Traxim dosisabhängig signifikant an (Tab. 1.). Auch beim Zinklaktat, das wegen Hinweisen auf geringere Verträglichkeit nur in der kleineren Dosis verabreicht wurde, kam es zu einem tendenziellen Anstieg des Zinkserumspiegels. Dagegen erfolgte nach Aufnahme von Zinkoxid in beiden Dosen so gut wie kein Zinkserumresponse. Tab. 1.: Mittelwerte der Maxima mit Standardabweichung, minimaler und maximaler Zinkserumwert beim Pony. (Mittelwerte die nicht mit demselben Buchstaben gekennzeichnet sind, unterscheiden sich signifikant). 10 mg Zn / kg KM 20 mg Zn / kg KM Verbindung Mittelwert der Maxima Stabw Range Mittelwert der Maxima Stabw Range Kontrolle 860 200b 660-1102 854 209b 649-1146 Zinkoxid 1100 340ab 776-1564 1136 463b 842-1785 Zinksulfat 2080 800a 1030-2805 4134 1323a 2880-5888 B-Traxim 2120 670a 1397-2885 4895 1497a 3136-6206 Zinklaktat 1810 630ab 1171-2638 • Nach zweiwöchiger Supplementierung von 2,5 mg Zink / kg KM zeigten sich bei den Ponys keine signifikanten Effekte auf den Zinkserumspiegel. Diese lagen innerhalb des in der Literatur beschriebenen Normalbereichs. • Bei den Hunden und Katzen kam es nach Verabreichung von 10 mg Zink /kg KM bei keinem der 4 Zinkpräparate zu einem signifikanten Anstieg des Serumzinkgehaltes. Allenfalls war bei den Katzen nach Aufnahme von Zinksulfat, B-Traxim und Zinklaktat eine gewissen Tendenz zu höheren Serumspiegeln zu erkennen. Aus den Untersuchungen kann geschlossen werden, dass Zinksulfat und B-Traxim beim Pferd verfügbar sein dürften, und sich daher für die Verwendung in Studien zu Zinksupplementation und Hufhornqualität eignen. Es kann aufgrund der eigenen Untersuchungen jedoch nicht ausgeschlossen werden, dass Zinkoxid und Zinklaktat nicht ebenfalls unter bestimmten Umständen beim Pferd verfügbar sind

Flow Field Estimation of Active Solute Transport – Information Transfer from Synthetic Data to Hele-Shaw Cell Experiments Using Convolutional Neural Networks

Variable density groundwater flow associated with active solute transport is understood reasonably well. Nevertheless, predictions are operationally still difficult due to joint effects of nonlinear processes and uncertain boundary conditions. Gaining deeper insight into the dynamics of these groundwater systems therefore relies on the availability of accurate and dense measurements of the complete system state and parameters. Often, such measurements are hard to come by, hence our information is incomplete. Recent deep learning methods in conjunction with numerical simulation of the physical processes to create large training datasets enable the information transfer to real world problems. To demonstrate this, I chose a laboratory experiment on density-driven active solute transport observed in a Hele-Shaw cell, where high resolution measurements of the solute concentration distribution are available. With the use of deep convolutional neural networks I was able to estimate the otherwise inaccessible flow fields and to identify the influence of background flow for this experiment without explicit knowledge of the boundary conditions. The situation of missing data, as encountered here, is typical also for other hydrological systems, from soil-vegetation-atmosphere interactions to catchment dynamics and groundwater recharge. Hence, I believe that the approach has wide applicability

Hematopoietic chimerism after allogeneic stem cell transplantation: a comparison of quantitative analysis by automated DNA sizing and fluorescent in situ hybridization

BACKGROUND: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is performed mainly in patients with high-risk or advanced hematologic malignancies and congenital or acquired aplastic anemias. In the context of the significant risk of graft failure after allo-HSCT from alternative donors and the risk of relapse in recipients transplanted for malignancy, the precise monitoring of posttransplant hematopoietic chimerism is of utmost interest. Useful molecular methods for chimerism quantification after allogeneic transplantation, aimed at distinguishing precisely between donor's and recipient's cells, are PCR-based analyses of polymorphic DNA markers. Such analyses can be performed regardless of donor's and recipient's sex. Additionally, in patients after sex-mismatched allo-HSCT, fluorescent in situ hybridization (FISH) can be applied. METHODS: We compared different techniques for analysis of posttransplant chimerism, namely FISH and PCR-based molecular methods with automated detection of fluorescent products in an ALFExpress DNA Sequencer (Pharmacia) or ABI 310 Genetic Analyzer (PE). We used Spearman correlation test. RESULTS: We have found high correlation between results obtained from the PCR/ALF Express and PCR/ABI 310 Genetic Analyzer. Lower, but still positive correlations were found between results of FISH technique and results obtained using automated DNA sizing technology. CONCLUSIONS: All the methods applied enable a rapid and accurate detection of post-HSCT chimerism

Cytotoxic Capacity of IL-15-Stimulated Cytokine-Induced Killer Cells Against Human Acute Myeloid Leukemia and Rhabdomyosarcoma in Humanized Preclinical Mouse Models

Allogeneic stem cell transplantation (allo-SCT) has become an important treatment modality for patients with high-risk acute myeloid leukemia (AML) and is also under investigation for soft tissue sarcomas. The therapeutic success is still limited by minimal residual disease (MRD) status ultimately leading to patients’ relapse. Adoptive donor lymphocyte infusions based on MRD status using IL-15-expanded cytokine-induced killer (CIK) cells may prevent relapse without causing graft-versus-host-disease (GvHD). To generate preclinical data we developed mouse models to study anti-leukemic- and anti-tumor-potential of CIK cells in vivo. Immunodeficient mice (NOD/SCID/IL-2Rγc−, NSG) were injected intravenously with human leukemic cell lines THP-1, SH-2 and with human rhabdomyosarcoma (RMS) cell lines RH41 and RH30 at minimal doses required for leukemia or tumor engraftment. Mice transplanted with THP-1 or RH41 cells were randomly assigned for analysis of CIK cell treatment. Organs of mice were analyzed by flow cytometry as well as quantitative polymerase chain reaction for engraftment of malignant cells and CIK cells. Potential of CIK cells to induce GvHD was determined by histological analysis. Tissues of the highest degree of THP-1 cell expansion included bone marrow followed by liver, lung, spleen, peripheral blood (PB), and brain. RH30 and RH41 engraftment mainly took place in liver and lung, but was also detectable in spleen and PB. In spite of delayed CIK cell expansion compared with malignant cells, CIK cells injected at equal amounts were sufficient for significant reduction of RH41 cells, whereas against fast-expanding THP-1 cells 250 times more CIK than THP-1 cells were needed to achieve comparable results. Our preclinical in vivo mouse models showed a reliable 100% engraftment of malignant cells which is essential for analysis of anti-cancer therapy. Furthermore our data demonstrated that IL-15-activated CIK cells have potent cytotoxic capacity against AML and RMS cells without causing GvHD

Epigenetic regulation of inflammation by microRNAs in post-infectious bronchiolitis obliterans

Post-infectious bronchiolitis obliterans (PiBO) is a rare, chronic disease initiated by severe infection and followed by perpetuating inflammation and obliteration of the small airways. MicroRNAs (miRNAs) have been proposed to play a central role as epigenetic regulators, which control resolution and prevent the uncontrolled progress of inflammation. The aim of this study was to define biomarkers on the level of post-transcriptional gene regulation in order to characterise PiBO. A total of 39 patients with well-defined PiBO and 31 controls from two centres, Barcelona, Spain, and Frankfurt, Germany, were analysed by next-generation sequencing (NGS). The evaluation of the biological targets of the miRNAs was performed by pathway enrichment analysis and protein-protein interaction network analysis respectively. Patients with PiBO had significantly lower lung function values and increased airway inflammation in induced sputum as indicated by total cell counts, neutrophils, IL-1β, IL-6, IL-8 and TGF-β compared to controls. Next-generation sequencing analysis revealed a total of 22 dysregulated miRNAs, which passed significance threshold for P adj ≤ 0.001 with 17 being upregulated and 5 being downregulated. Of these dysregulated miRNAs, miR-335-5p, miR-186-5p, miR-30b-5p and miR-30c-5p were further validated using qRT-PCR. Interestingly, these miRNAs are functionally implicated in cytokine-cytokine receptor interaction, TGF-β signalling and FoxO signalling pathway and significantly correlated with lung function values (FEV1). Our results demonstrate an aberrant miRNA expression profile in PiBO, which impacts pathways responsible for the regulation of inflammation and fibrosis. The defined miRNAs are useful biomarkers and should be assessed as potential target in the field of miRNA therapeutics. We identified dysregulated miRNAs, which impact pathways for inflammatory cytokines and TGF-β signalling in post-infectious bronchiolitis obliterans. The miRNAs reflect bronchial inflammation and fibrosis and could be considered as novel biomarkers supporting diagnosis and treatment options

Epigenetic regulation of inflammation by microRNAs in post-infectious bronchiolitis obliterans

Post-infectious bronchiolitis obliterans (PiBO) is a rare, chronic disease initiated by severe infection and followed by perpetuating inflammation and obliteration of the small airways. MicroRNAs (miRNAs) have been proposed to play a central role as epigenetic regulators, which control resolution and prevent the uncontrolled progress of inflammation. The aim of this study was to define biomarkers on the level of post-transcriptional gene regulation in order to characterise PiBO. A total of 39 patients with well-defined PiBO and 31 controls from two centres, Barcelona, Spain, and Frankfurt, Germany, were analysed by next-generation sequencing (NGS). The evaluation of the biological targets of the miRNAs was performed by pathway enrichment analysis and protein-protein interaction network analysis respectively. Patients with PiBO had significantly lower lung function values and increased airway inflammation in induced sputum as indicated by total cell counts, neutrophils, IL-1β, IL-6, IL-8 and TGF-β compared to controls. Next-generation sequencing analysis revealed a total of 22 dysregulated miRNAs, which passed significance threshold for P adj ≤ 0.001 with 17 being upregulated and 5 being downregulated. Of these dysregulated miRNAs, miR-335-5p, miR-186-5p, miR-30b-5p and miR-30c-5p were further validated using qRT-PCR. Interestingly, these miRNAs are functionally implicated in cytokine-cytokine receptor interaction, TGF-β signalling and FoxO signalling pathway and significantly correlated with lung function values (FEV1). Our results demonstrate an aberrant miRNA expression profile in PiBO, which impacts pathways responsible for the regulation of inflammation and fibrosis. The defined miRNAs are useful biomarkers and should be assessed as potential target in the field of miRNA therapeutics. We identified dysregulated miRNAs, which impact pathways for inflammatory cytokines and TGF-β signalling in post-infectious bronchiolitis obliterans. The miRNAs reflect bronchial inflammation and fibrosis and could be considered as novel biomarkers supporting diagnosis and treatment options

Epigenetic regulation of inflammation by microRNAs in post-infectious bronchiolitis obliterans

Post-infectious bronchiolitis obliterans (PiBO) is a rare, chronic disease initiated by severe infection and followed by perpetuating inflammation and obliteration of the small airways. MicroRNAs (miRNAs) have been proposed to play a central role as epigenetic regulators, which control resolution and prevent the uncontrolled progress of inflammation. The aim of this study was to define biomarkers on the level of post-transcriptional gene regulation in order to characterise PiBO. A total of 39 patients with well-defined PiBO and 31 controls from two centres, Barcelona, Spain, and Frankfurt, Germany, were analysed by next-generation sequencing (NGS). The evaluation of the biological targets of the miRNAs was performed by pathway enrichment analysis and protein-protein interaction network analysis respectively. Patients with PiBO had significantly lower lung function values and increased airway inflammation in induced sputum as indicated by total cell counts, neutrophils, IL-1β, IL-6, IL-8 and TGF-β compared to controls. Next-generation sequencing analysis revealed a total of 22 dysregulated miRNAs, which passed significance threshold for P adj ≤ 0.001 with 17 being upregulated and 5 being downregulated. Of these dysregulated miRNAs, miR-335-5p, miR-186-5p, miR-30b-5p and miR-30c-5p were further validated using qRT-PCR. Interestingly, these miRNAs are functionally implicated in cytokine-cytokine receptor interaction, TGF-β signalling and FoxO signalling pathway and significantly correlated with lung function values (FEV1). Our results demonstrate an aberrant miRNA expression profile in PiBO, which impacts pathways responsible for the regulation of inflammation and fibrosis. The defined miRNAs are useful biomarkers and should be assessed as potential target in the field of miRNA therapeutics. We identified dysregulated miRNAs, which impact pathways for inflammatory cytokines and TGF-β signalling in post-infectious bronchiolitis obliterans. The miRNAs reflect bronchial inflammation and fibrosis and could be considered as novel biomarkers supporting diagnosis and treatment options