37 results on '"Grazyna Domanska"'
Search Results
2. Increased mortality and altered local immune response in secondary peritonitis after previous visceral operations in mice
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Jonas Menz, Laura Hundt, Tobias Schulze, Katrin Schmoeckel, Pia Menges, and Grazyna Domanska
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Medicine ,Science - Abstract
Abstract Postoperative peritonitis is characterized by a more severe clinical course than other forms of secondary peritonitis. The pathophysiological mechanisms behind this phenomenon are incompletely understood. This study used an innovative model to investigate these mechanisms, combining the models of murine Colon Ascendens Stent Peritonitis (CASP) and Surgically induced Immune Dysfunction (SID). Moreover, the influence of the previously described anti-inflammatory reflex transmitted by the vagal nerve was characterized. SID alone, or 3 days before CASP were performed in female C57BL/6 N mice. Subdiaphragmatic vagotomy was performed six days before SID with following CASP. The immune status was assessed by FACS analysis and measurement of cytokines. Local intestinal inflammatory changes were characterized by immunohistochemistry. Mortality was increased in CASP animals previously subjected to SID. Subclinical bacteremia occurred after SID, and an immunosuppressive milieu occurred secondary to SID just before the induction of CASP. Previous SID modified the pattern of intestinal inflammation induced by CASP. Subdiaphragmatic vagotomy had no influence on sepsis mortality in our model of postoperative peritonitis. Our results indicate a surgery-induced inflammation of the small intestine and the peritoneal cavity with bacterial translocation, which led to immune dysfunction and consequently to a more severe peritonitis.
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- 2021
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3. Immune Polarization Potential of the S. aureus Virulence Factors SplB and GlpQ and Modulation by Adjuvants
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Daniel M. Mrochen, Patricia Trübe, Ilka Jorde, Grazyna Domanska, Cindy van den Brandt, and Barbara M. Bröker
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Staphylococcus aureus ,vaccine ,adjuvants ,SplB ,GlpQ ,immune polarization ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Protection against Staphylococcus aureus is determined by the polarization of the anti-bacterial immune effector mechanisms. Virulence factors of S. aureus can modulate these and induce differently polarized immune responses in a single individual. We proposed that this may be due to intrinsic properties of the bacterial proteins. To test this idea, we selected two virulence factors, the serine protease-like protein B (SplB) and the glycerophosphoryl diester phosphodiesterase (GlpQ). In humans naturally exposed to S. aureus, SplB induces a type 2-biased adaptive immune response, whereas GlpQ elicits type 1/type 3 immunity. We injected the recombinant bacterial antigens into the peritoneum of S. aureus-naïve C57BL/6N mice and analyzed the immune response. This was skewed by SplB toward a Th2 profile including specific IgE, whereas GlpQ was weakly immunogenic. To elucidate the influence of adjuvants on the proteins’ polarization potential, we studied Montanide ISA 71 VG and Imject™Alum, which promote a Th1 and Th2 response, respectively. Alum strongly increased antibody production to the Th2-polarizing protein SplB, but did not affect the response to GlpQ. Montanide enhanced the antibody production to both S. aureus virulence factors. Montanide also augmented the inflammation in general, whereas Alum had little effect on the cellular immune response. The adjuvants did not override the polarization potential of the S. aureus proteins on the adaptive immune response.
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- 2021
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4. The Immunomodulator 1-Methyltryptophan Drives Tryptophan Catabolism Toward the Kynurenic Acid Branch
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Elisa Wirthgen, Anne K. Leonard, Christian Scharf, and Grazyna Domanska
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1-MT ,IDO ,KYNA ,kynurenine pathway ,tryptophan ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Background: Animal model studies revealed that the application of 1-methyltryptophan (1-MT), a tryptophan (TRP) analog, surprisingly increased plasma levels of the TRP metabolite, kynurenic acid (KYNA). Under inflammatory conditions, KYNA has been shown to mediate various immunomodulatory effects. Therefore, the present study aims to confirm and clarify the effects of 1-MT on TRP metabolism in mice as well as in humans.Methods: Splenocytes from Balb/C or indoleamine 2,3-dioxygenase knockout (IDO1−/−) mice or whole human blood were stimulated with 1-MT for 6, 24, or 36 h. C57BL/6 mice received 1-MT in drinking water for 5 days. Cell-free supernatants and plasma were analyzed for TRP and its metabolites by tandem mass spectrometry (MS/MS).Results: 1-MT treatment induced an increase in TRP and its metabolite, KYNA in Balb/C, IDO−/− mice, and in human blood. Concurrently, the intermediate metabolite kynurenine (KYN), as well as the KYN/TRP ratio, were reduced after 1-MT treatment. The effects of 1-MT on TRP metabolites were similar after the in vivo application of 1-MT to C57BL/6 mice.Conclusions: The data indicate that 1-MT induced an increase of KYNA ex vivo and in vivo confirming previously described results. Furthermore, the results of IDO−/− mice indicate that this effect seems not to be mediated by IDO1. Due to the proven immunomodulatory properties of KYNA, a shift toward this branch of the kynurenine pathway (KP) may be one potential mode of action by 1-MT and should be considered for further applications.
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- 2020
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5. Activation of the Kynurenine Pathway in Human Malignancies Can Be Suppressed by the Cyclin-Dependent Kinase Inhibitor Dinaciclib
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Christin Riess, Björn Schneider, Hanna Kehnscherper, Julia Gesche, Nina Irmscher, Fatemeh Shokraie, Carl Friedrich Classen, Elisa Wirthgen, Grazyna Domanska, Annette Zimpfer, Daniel Strüder, Christian Junghanss, and Claudia Maletzki
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targeted therapy ,solid tumor models ,tryptophan metabolites ,IDO1 ,chemotherapy ,Immunologic diseases. Allergy ,RC581-607 - Abstract
Indoleamine 2,3-dioxygenase (IDO) and tryptophan 2,3-dioxygenase (TDO2) are the key enzymes of tryptophan (TRP) metabolism in the kynurenine pathway (KP). Both enzymes function as indicators of immunosuppression and poor survival in cancer patients. Direct or indirect targeting of either of these substances seems thus reasonable to improve therapy options for patients. In this study, glioblastoma multiforme (GBM) as well as head and neck squamous cell carcinomas (HNSCC) were examined because of their different mechanisms of spontaneous and treatment-induced immune escape. Effects on gene expression and protein levels were examined. Accompanying assessment of TRP metabolites from treated GBM cell culture supernatants was conducted. Our results show a heterogeneous and inversely correlated expression profile of TRP-metabolizing genes among GBM and HNSCC cells, with low, but inducible IDO1 expression upon IFNγ treatment. TDO2 expression was higher in GBM cells, while genes encoding kynurenine aminotransferases were mainly confined to HNSCC cells. These data indicate that the KP is active in both entities, with however different enzymes involved in TRP catabolism. Upon treatment with Temozolomide, the standard of care for GBM patients, IDO1 was upregulated. Comparable, although less pronounced effects were seen in HNSCC upon Cetuximab and conventional drugs (i.e., 5-fluorouracil, Gemcitabine). Here, IDO1 and additional genes of the KP (KYAT1, KYAT2, and KMO) were induced. Vice versa, the novel yet experimental cyclin-dependent kinase inhibitor Dinaciclib suppressed KP in both entities. Our comprehensive data imply inhibition of the TRP catabolism by Dinaciclib, while conventional chemotherapeutics tend to activate this pathway. These data point to limitations of conventional therapy and highlight the potential of targeted therapies to interfere with the cells' metabolism more than anticipated.
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- 2020
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6. Obesity Impairs Mobility and Adult Hippocampal Neurogenesis
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Alexander Bracke, Grazyna Domanska, Katharina Bracke, Steffen Harzsch, Jens van den Brandt, Barbara Bröker, and Oliver von Bohlen und Halbach
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Neurosciences. Biological psychiatry. Neuropsychiatry ,RC321-571 - Abstract
Currently, it is controversially discussed whether a relationship between obesity and cognition exists. We here analyzed a mouse model of obesity (leptin-deficient mice) to study the effects of obesity on the morphology of the hippocampus (a brain structure involved in mechanisms related to learning and memory) and on behavior. Mice aged 4 to 6 months were analyzed. At this age, the obese mice have nearly double the body weight as controls, but display smaller brains (brain volume is about 10% smaller) as control animals of the same age. Adult hippocampal neurogenesis, a process that is linked to learning and memory, might be disturbed in the obese mice and contribute to the smaller brain volume. Adult hippocampal neurogenesis was examined using specific markers for cell proliferation (phosphohistone H3), neuronal differentiation (doublecortin), and apoptosis (caspase 3). The number of phosphohistone H3 and doublecortin-positive cells was markedly reduced in leptin-deficient mice, but not the number of apoptotic cells, indicating that adult hippocampal neurogenesis on the level of cell proliferation was affected. In addition, dendritic spine densities of pyramidal neurons in the hippocampal area CA1 were analyzed using Golgi impregnation. However, no significant change in dendritic spine densities was noted in the obese mice. Moreover, the performance of the mice was analyzed in the open field as well as in the Morris water maze. In the open field test, obese mice showed reduced locomotor activity, but in the Morris water maze they showed similar performance compared with control animals.
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- 2019
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7. The Impact of Lidocaine on Adipose-Derived Stem Cells in Human Adipose Tissue Harvested by Liposuction and Used for Lipotransfer
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Felix Grambow, Rico Rutkowski, Fred Podmelle, Katrin Schmoeckel, Florian Siegerist, Grzegorz Domanski, Matthias W. Schuster, and Grazyna Domanska
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lipofilling ,autologous lipotransfer ,adipose-derived stem cells ,lidocaine ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
The local anesthetic lidocaine, which has been used extensively during liposuction, has been reported to have cytotoxic effects and therefore would be unsuitable for use in autologous lipotransfer. We evaluated the effect of lidocaine on the distribution, number, and viability of adipose-derived stem cells (ASCs), preadipocytes, mature adipocytes, and leukocytes in the fatty and fluid portion of the lipoaspirate using antibody staining and flow cytometry analyses. Adipose tissue was harvested from 11 female patients who underwent liposuction. Abdominal subcutaneous fat tissue was infiltrated with tumescent local anesthesia, containing lidocaine on the left and lacking lidocaine on the right side of the abdomen, and harvested subsequently. Lidocaine had no influence on the relative distribution, cell number, or viability of ASCs, preadipocytes, mature adipocytes, or leukocytes in the stromal-vascular fraction. Assessing the fatty and fluid portions of the lipoaspirate, the fatty portions contained significantly more ASCs (p < 0.05), stem cells expressing the preadipocyte marker Pref-1 (p < 0.01 w/lidocaine, p < 0.05 w/o lidocaine), and mature adipocytes (p < 0.05 w/lidocaine, p < 0.01 w/o lidocaine) than the fluid portions. Only the fatty portion should be used for transplantation. This study found no evidence that would contraindicate the use of lidocaine in lipotransfer. Limitations of the study include the small sample size and the inclusion of only female patients.
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- 2020
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8. Effects of 1-Methyltryptophan on Immune Responses and the Kynurenine Pathway after Lipopolysaccharide Challenge in Pigs
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Elisa Wirthgen, Winfried Otten, Margret Tuchscherer, Armin Tuchscherer, Grazyna Domanska, Julia Brenmoehl, Juliane Günther, Daniela Ohde, Werner Weitschies, Anne Seidlitz, Eberhard Scheuch, and Ellen Kanitz
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indoleamine 2,3-dioxygenase ,kynurenine pathway ,methyltryptophan ,LPS ,pig ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
An enhanced indoleamine 2,3-dioxygenase 1 (IDO1) activity is associated with an increased mortality risk in sepsis patients. Thus, the preventive inhibition of IDO1 activity may be a promising strategy to attenuate the severity of septic shock. 1-methyltryptophan (1-MT) is currently in the interest of research due to its potential inhibitory effects on IDO1 and immunomodulatory properties. The present study aims to investigate the protective and immunomodulatory effects of 1-methyltryptophan against endotoxin-induced shock in a porcine in vivo model. Effects of 1-MT were determined on lipopolysaccharide (LPS)-induced tryptophan (TRP) degradation, immune response and sickness behaviour. 1-MT increased TRP and its metabolite kynurenic acid (KYNA) in plasma and tissues, suppressed the LPS-induced maturation of neutrophils and increased inactivity of the animals. 1-MT did not inhibit the LPS-induced degradation of TRP to kynurenine (KYN)—a marker for IDO1 activity—although the increase in KYNA indicates that degradation to one branch of the KYN pathway is facilitated. In conclusion, our findings provide no evidence for IDO1 inhibition but reveal the side effects of 1-MT that may result from the proven interference of KYNA and 1-MT with aryl hydrocarbon receptor signalling. These effects should be considered for therapeutic applications of 1-MT.
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- 2018
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9. Psychological stress-induced, IDO1-dependent tryptophan catabolism: implications on immunosuppression in mice and humans.
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Cornelia Kiank, Jan-Philip Zeden, Solveig Drude, Grazyna Domanska, Gerhard Fusch, Winfried Otten, and Christine Schuett
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Medicine ,Science - Abstract
It is increasingly recognized that psychological stress influences inflammatory responses and mood. Here, we investigated whether psychological stress (combined acoustic and restraint stress) activates the tryptophan (Trp) catabolizing enzyme indoleamine 2,3-dioxygenase 1(IDO1) and thereby alters the immune homeostasis and behavior in mice. We measured IDO1 mRNA expression and plasma levels of Trp catabolites after a single 2-h stress session and in repeatedly stressed (4.5-days stress, 2-h twice a day) naïve BALB/c mice. A role of cytokines in acute stress-induced IDO1 activation was studied after IFNgamma and TNFalpha blockade and in IDO1(-/-) mice. RU486 and 1-Methyl-L-tryptophan (1-MT) were used to study role of glucocorticoids and IDO1 on Trp depletion in altering the immune and behavioral response in repeatedly stressed animals. Clinical relevance was addressed by analyzing IDO1 activity in patients expecting abdominal surgery. Acute stress increased the IDO1 mRNA expression in brain, lung, spleen and Peyer's patches (max. 14.1+/-4.9-fold in brain 6-h after stress) and resulted in a transient depletion of Trp (-25.2+/-6.6%) and serotonin (-27.3+/-4.6%) from the plasma measured 6-h after stress while kynurenine levels increased 6-h later (11.2+/-9.3%). IDO1 mRNA up-regulation was blocked by anti-TNFalpha and anti-IFNgamma treatment. Continuous IDO1 blockade by 1-MT but not RU486 treatment normalized the anti-bacterial defense and attenuated increased IL-10 inducibility in splenocytes after repeated stress as it reduced the loss of body weight and behavioral alterations. Moreover, kynurenic acid which remained increased in 1-MT treated repeatedly stressed mice was identified to reduce the TNFalpha inducibility of splenocytes in vitro and in vivo. Thus, psychological stress stimulates cytokine-driven IDO1 activation and Trp depletion which seems to have a central role for developing stress-induced immunosuppression and behavioral alteration. Since patients showed Trp catabolism already prior to surgery, IDO is also a possible target enzyme for humans modulating immune homeostasis and mood.
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- 2010
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10. Association between waist circumference and gray matter volume in 2344 individuals from two adult community-based samples.
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Deborah Janowitz, Katharina Wittfeld, Jan Terock, Harald Jürgen Freyberger, Katrin Hegenscheid, Henry Völzke, Mohamad Habes, Norbert Hosten, Nele Friedrich, Matthias Nauck, Grazyna Domanska, and Hans Jörgen Grabe
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- 2015
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11. Increased mortality and altered local immune response in secondary peritonitis after previous visceral operations in mice
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Grazyna Domanska, P. Menges, Katrin Schmoeckel, Jonas Menz, Laura Hundt, and Tobias Schulze
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medicine.medical_specialty ,Science ,Immunology ,Peritonitis ,Inflammation ,Pathogenesis ,Gastroenterology ,Article ,Mice ,Peritoneal cavity ,Postoperative Complications ,Immune system ,Internal medicine ,medicine ,Animals ,CASP ,Peritoneal Cavity ,Subclinical infection ,Multidisciplinary ,business.industry ,Immunity ,medicine.disease ,humanities ,Pathophysiology ,Experimental models of disease ,Disease Models, Animal ,medicine.anatomical_structure ,Bacteremia ,Medicine ,medicine.symptom ,business - Abstract
Postoperative peritonitis is characterized by a more severe clinical course than other forms of secondary peritonitis. The pathophysiological mechanisms behind this phenomenon are incompletely understood. This study used an innovative model to investigate these mechanisms, combining the models of murine Colon Ascendens Stent Peritonitis (CASP) and Surgically induced Immune Dysfunction (SID). Moreover, the influence of the previously described anti-inflammatory reflex transmitted by the vagal nerve was characterized. SID alone, or 3 days before CASP were performed in female C57BL/6 N mice. Subdiaphragmatic vagotomy was performed six days before SID with following CASP. The immune status was assessed by FACS analysis and measurement of cytokines. Local intestinal inflammatory changes were characterized by immunohistochemistry. Mortality was increased in CASP animals previously subjected to SID. Subclinical bacteremia occurred after SID, and an immunosuppressive milieu occurred secondary to SID just before the induction of CASP. Previous SID modified the pattern of intestinal inflammation induced by CASP. Subdiaphragmatic vagotomy had no influence on sepsis mortality in our model of postoperative peritonitis. Our results indicate a surgery-induced inflammation of the small intestine and the peritoneal cavity with bacterial translocation, which led to immune dysfunction and consequently to a more severe peritonitis.
- Published
- 2021
12. Siponimod (BAF312) Treatment Reduces Brain Infiltration but Not Lesion Volume in Middle-Aged Mice in Experimental Stroke
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Grazyna Domanska, Michael Kirsch, Juliane Schulze, Johanna Ruhnau, Antje Vogelgesang, and Alexander Dressel
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Male ,Sphingosine 1 Phosphate Receptor Modulators ,Pathology ,medicine.medical_specialty ,T-Lymphocytes ,Central nervous system ,Ischemia ,Brain Ischemia ,Lesion ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Benzyl Compounds ,medicine ,Animals ,Neuroinflammation ,030304 developmental biology ,Advanced and Specialized Nursing ,0303 health sciences ,business.industry ,Multiple sclerosis ,Age Factors ,Brain ,medicine.disease ,Pathophysiology ,Mice, Inbred C57BL ,Stroke ,Disease Models, Animal ,Treatment Outcome ,Lymphatic system ,medicine.anatomical_structure ,Siponimod ,chemistry ,Azetidines ,Neurology (clinical) ,medicine.symptom ,Cardiology and Cardiovascular Medicine ,business ,030217 neurology & neurosurgery - Abstract
Background and Purpose— The contribution of neuroinflammation and, in particular, the infiltration of the brain by lymphocytes is increasingly recognized as a substantial pathophysiological mechanism after stroke. The interaction of lymphocytes with endothelial cells and platelets, termed thromboinflammation, fosters microvascular dysfunction and secondary infarct growth. Siponimod is an S1PR (sphingosine-1-phosphate receptor) modulator, which blocks the egress of lymphocytes from lymphoid organs and has demonstrated beneficial effects in multiple sclerosis treatment. We investigated the effect of treatment with siponimod on stroke outcome in a mouse model of cerebral ischemia. Methods— Transient middle cerebral artery occlusion was induced in middle-aged wild-type mice. Animals were either treated with siponimod (3 mg/kg; intraperitoneal) or vehicle for 6 days. Stroke outcome was assessed by magnetic resonance imaging (spleen volume: prestroke, day 3, and day 7; infarct volume: days 1, 3, and 7) and behavioral tests (prestroke, day 2, and day 6). Immune cells of the peripheral blood and brain-infiltrating cells ipsilateral and contralateral were analyzed by VETScan and by flow cytometry. Results— Siponimod significantly induced lymphopenia on day 7 after transient middle cerebral artery occlusion and reduced T-lymphocyte accumulation in the central nervous system. No effect was detected for lesion size. Conclusions— For siponimod administered at 3 mg/kg in transient middle cerebral artery occlusion mouse model, our findings do not provide preclinical evidence for the use of S1PR1/5 modulators as neuroprotectant in stroke therapy.
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- 2019
13. Immune Polarization Potential of the S. aureus Virulence Factors SplB and GlpQ and Modulation by Adjuvants
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Ilka Jorde, Daniel M. Mrochen, Grazyna Domanska, Barbara M. Bröker, Cindy van den Brandt, and Patricia Trübe
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lcsh:Immunologic diseases. Allergy ,0301 basic medicine ,Staphylococcus aureus ,Virulence Factors ,030106 microbiology ,Immunology ,Antigen-Presenting Cells ,Virulence ,chemical and pharmacologic phenomena ,Inflammation ,medicine.disease_cause ,Immunoglobulin E ,Microbiology ,Th2 ,Mice ,03 medical and health sciences ,Th2 Cells ,Immune system ,Adjuvants, Immunologic ,Bacterial Proteins ,Immunity ,vaccine ,medicine ,Animals ,Immunology and Allergy ,mouse models ,GlpQ ,Original Research ,Antigens, Bacterial ,biology ,Phosphoric Diester Hydrolases ,Chemistry ,SplB ,Th1 Cells ,Acquired immune system ,Antibodies, Bacterial ,Mice, Inbred C57BL ,030104 developmental biology ,adjuvants ,biology.protein ,Cytokines ,immune polarization ,Female ,Immunization ,Bacterial antigen ,medicine.symptom ,lcsh:RC581-607 - Abstract
Protection against Staphylococcus aureus is determined by the polarization of the anti-bacterial immune effector mechanisms. Virulence factors of S. aureus can modulate these and induce differently polarized immune responses in a single individual. We proposed that this may be due to intrinsic properties of the bacterial proteins. To test this idea, we selected two virulence factors, the serine protease-like protein B (SplB) and the glycerophosphoryl diester phosphodiesterase (GlpQ). In humans naturally exposed to S. aureus, SplB induces a type 2-biased adaptive immune response, whereas GlpQ elicits type 1/type 3 immunity. We injected the recombinant bacterial antigens into the peritoneum of S. aureus-naïve C57BL/6N mice and analyzed the immune response. This was skewed by SplB toward a Th2 profile including specific IgE, whereas GlpQ was weakly immunogenic. To elucidate the influence of adjuvants on the proteins’ polarization potential, we studied Montanide ISA 71 VG and Imject™Alum, which promote a Th1 and Th2 response, respectively. Alum strongly increased antibody production to the Th2-polarizing protein SplB, but did not affect the response to GlpQ. Montanide enhanced the antibody production to both S. aureus virulence factors. Montanide also augmented the inflammation in general, whereas Alum had little effect on the cellular immune response. The adjuvants did not override the polarization potential of the S. aureus proteins on the adaptive immune response.
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- 2021
14. Vitamin B6 deficiency in new born rats affects hepatic cardiolipin composition and oxidative phosphorylation
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Uwe Lendeckel, Grazyna Domanska, Daniela Peter, Lorenz Schild, Jens Weingärtner, Sarah Gürtler, and Carmen Wolke
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0301 basic medicine ,Male ,medicine.medical_specialty ,Cardiolipins ,Mitochondria, Liver ,Oxidative phosphorylation ,01 natural sciences ,General Biochemistry, Genetics and Molecular Biology ,Oxidative Phosphorylation ,03 medical and health sciences ,chemistry.chemical_compound ,Oxygen Consumption ,Pregnancy ,Internal medicine ,Cardiolipin ,medicine ,Animals ,Original Research ,010401 analytical chemistry ,Plasma levels ,medicine.disease ,Vitamin D Deficiency ,Mitochondrial respiration ,0104 chemical sciences ,Rats ,030104 developmental biology ,Endocrinology ,chemistry ,Animals, Newborn ,Liver ,Rats, Inbred Lew ,Composition (visual arts) ,Female ,Vitamin b6 - Abstract
Vitamin B6 deficiency during pregnancy translates into a severe vitamin B6 deficiency (plasma levels decreased by 97%) in new-born rats. Further, hallmarks are increased (+89%) concentrations of homocysteine, gross changes in gene methylation and expression, and metabolic alterations including lipid metabolism. This study focuses on determining the effects of vitamin B6-deficiency on cardiolipin composition and oxidative phosphorylation in liver. For this purpose, hepatic cardiolipin composition was analyzed by means of LC/MS/MS, and mitochondrial oxygen consumption was determined by using a Clark-type electrode in a rat model of vitamin B6 deficiency. Liver mitochondria from new-born rats with pre-term vitamin B6 deficiency responded with substantial alterations in cardiolipin composition that include the following changes in the amounts of cardiolipin incorporated fatty acids: increase in C16, decrease in C18, decrease in saturated fatty acid, as well as increase in amount of oxidized cardiolipin species. These changes were accompanied by significantly decreased capacity of oxidative phosphorylation. In conclusion, vitamin B6 deficiency in new born rats induces massive alterations of cardiolipin composition and function of liver mitochondria. These findings support the importance of sufficient periconceptional supply of vitamin B6 to prevent vitamin B6 deficiency.Impact statementVitamin B6 (VitB6) is an active co-enzyme for more than 150 enzymes and is required for a great diversity of biosynthesis and metabolic reactions. There is an increased need for VitB6 during pregnancy and sufficient supply of VitB6 is crucial for the prevention of cleft palate and neural tube defects. We show that liver mitochondria from new-born rats with pre-term VitB6 deficiency respond with substantial alterations in cardiolipin (CL) composition and in the amount of oxidized CL species. These changes are associated with a decrease in the efficiency of oxidative phosphorylation. The results of this study support the significance of sufficient supply of VitB6 during pregnancy (and periconceptional) for diminishing the number of early abortions and minimizing malformation. The established link between VitB6 deficiency, CL composition, and mitochondrial respiration/energy production provides mechanistic insight as to how the VitB6 deficiency translates into the known pathophysiological and clinically relevant conditions.
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- 2019
15. Obesity alters mobility and adult neurogenesis, but not hippocampal dependent learning in ob/ob mice
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Katharina Bracke, von Bohlen und Halbach O, van den Brandt J, Barbara M. Bröker, Alexander Bracke, Steffen Harzsch, and Grazyna Domanska
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medicine.medical_specialty ,education.field_of_study ,Dentate gyrus ,Population ,Neurogenesis ,Morris water navigation task ,Hippocampus ,Hippocampal formation ,Biology ,Cell morphology ,Doublecortin ,Endocrinology ,Internal medicine ,medicine ,biology.protein ,education - Abstract
sBackgroundObesity has become a severe problem among the world’s population with clearly increasing prevalence over the last decades. Because obesity is associated with several comorbidities (e.g. hypertension or cancer) it constitutes an increasing burden for the health care system. Correlations between obesity and cognition have been studied in humans with ambivalent results. Here, we studied the effects of obesity on hippocampus dependent learning and memory and cell morphology in a mouse model of obesity.MethodsThe body mass of male and female Lep+/+(wt) and Lepob/ob(ob/ob) animals with access to food and water ad libitum was measured between postnatal day 60-200 and animals with clear adiposity (4-6 months) were further analyzed. Adult hippocampal neurogenesis in the dentate gyrus was examined using phosphohistone H3 as a marker for proliferation, doublecortin as a marker for differentiation and caspase3 as a marker for apoptosis. Moreover, the density of dendritic spines on apical and basal dendrites of pyramidal neurons of the cornu ammonis 1 (CA1) were analyzed using Golgi impregnation. In addition, mice were subjected to the open field and Morris water maze test in order to analyze locomotor activity and spatial learning.ResultsThe body weight of ob/ob mice nearly doubled during the first 120 postnatal days. Adult hippocampal neurogenesis was reduced in ob/ob mice due to reduced cell proliferation. Dendritic spine densities in the hippocampal area CA1 were not altered in ob/ob mice. Four to six months old ob/ob mice showed reduced locomotor activity in the open field test but similar performance in the Morris water maze compared to control mice.ConclusionOur data show that alterations in adult neurogenesis in leptin-deficient mice are not associated with an impairment in spatial learning abilities. Moreover, ob/ob mice are inconspicuous in the Morris water maze and do not display altered spine densities in the hippocampus, suggesting that obesity does not have a severe impact upon hippocampal neuronal plasticity and spatial learning.
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- 2019
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16. Simulation of the Measured Reactivity Distributions in the Subcritical MYRRHA Reactor
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Jerzy Janczyszyn, Grażyna Domańska, and Mikołaj Oettingen
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ADS ,MYRRHA ,reactivity ,detectors ,spatial effect ,area method ,Technology - Abstract
The designed MYRRHA reactor, in its subcritical version, will be equipped with a set of detectors monitoring its condition by measuring the current value of negative reactivity, which is a crucial parameter for its safe operation. In subcritical systems, accurate and precise measurement of negative reactivity is disturbed by the so-called spatial effect, i.e., the response of detectors depends on their placement in the reactor core. This paper focuses on the Monte Carlo simulations of reactivity measurements using the area method for natU, 238U, 241Am, 239Pu, and 232Th detectors. The simulations were performed in six positions with increasing distance from the center of the core and at three axial levels. The obtained results allow for selecting optimum locations for detectors and detector nuclides in terms of the accuracy of reactivity measurement and illustrate the dependence of the reactivity on the distance. Additionally, the possibility of using 103Rh in self-powered neutron detectors was investigated. The influence of spatial effect in calculations using the area method was directly indicated in the MYRRHA reactor core for chosen isotopes and in-core positions. The results closest to true values were obtained for the second fuel assembly for 239Pu, and the third fuel assembly for natU, 238U, 232Th, and 241Am; thus, these nuclides and positions should be preferred when selecting detectors for MYRRHA.
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- 2024
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17. Altered hepatic mRNA expression of immune response and apoptosis-associated genes after acute and chronic psychological stress in mice
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Maren Depke, Uwe Völker, Cornelia Kiank, Leif Steil, Christine Schütt, and Grazyna Domanska
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Cell signaling ,Programmed cell death ,Immunology ,Antigen presentation ,Gene Expression ,Apoptosis ,Biology ,medicine.disease_cause ,Protein Carbonylation ,Mice ,Immune system ,Cell Movement ,Gene expression ,medicine ,Animals ,RNA, Messenger ,Molecular Biology ,Oligonucleotide Array Sequence Analysis ,Mice, Inbred BALB C ,Gene Expression Profiling ,Up-Regulation ,Gene expression profiling ,Oxidative Stress ,Liver ,Acute Disease ,Chronic Disease ,Female ,Stress, Psychological ,Oxidative stress - Abstract
Using a combination of transcriptional profiling and Ingenuity Pathway Analysis (IPA, www.ingenuity.com) we investigated acute and chronic psychological stress induced alterations of hepatic gene expression of BALB/c mice. Already after a 2-h single stress session, up-regulation of several LPS and glucocorticoid-sensitive immune response genes and markers related to oxidative stress and apoptotic processes were observed. Support for the existence of oxidative stress was gained by measuring increased protein carbonylation, but no alterations of immune responsiveness or cell death were measured in mice after acute stress compared to the control group. When animals were repeatedly stressed during 4.5-days, we found reduced transcription of antigen presentation molecules, altered mRNA levels of immune cell signaling mediators and persisting high expression of apoptosis-related genes. These alterations were associated with a measurable immune suppression characterized by a reduced ability to clear experimental Salmonella typhimurium infection from the liver and a heightened hepatocyte apoptosis. Moreover, genes associated with anti-oxidative functions and regenerative processes were induced in the hepatic tissue of chronically stressed mice. These findings indicate that modulation of the immune response and of apoptosis-related genes is initiated already during a single acute stress exposure. However, immune suppression will only manifest in repeatedly stressed mice which additionally show induction of protective and liver regenerative genes to prevent further hepatocyte damage.
- Published
- 2009
18. Pharmacokinetics of 1-methyl-L-tryptophan after single and repeated subcutaneous application in a porcine model
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Margret Tuchscherer, Ellen Kanitz, Grazyna Domanska, Anne Seidlitz, Elisa Wirthgen, Winfried Otten, Werner Weitschies, Armin Tuchscherer, and Eberhard Scheuch
- Subjects
0301 basic medicine ,Single administration ,pig ,medicine.medical_specialty ,Time Factors ,Original ,Swine ,Injections, Subcutaneous ,methyltryptophan ,General Biochemistry, Genetics and Molecular Biology ,Subcutaneous application ,03 medical and health sciences ,Pharmacokinetics ,Internal medicine ,medicine ,Animals ,Indoleamine-Pyrrole 2,3,-Dioxygenase ,Tissue Distribution ,Enzyme Inhibitors ,Indoleamine 2,3-dioxygenase ,chemistry.chemical_classification ,General Veterinary ,biology ,Tryptophan ,General Medicine ,pharmacokinetics ,indoleamine 2,3-dioxygenase ,tryptophan ,Enzyme assay ,Sus scrofa domestica ,030104 developmental biology ,Endocrinology ,Enzyme ,3-dioxygenase ,chemistry ,Models, Animal ,biology.protein ,indoleamine 2 ,Animal Science and Zoology - Abstract
Increased activity of the tryptophan-metabolizing enzyme indoleamine 2,3-dioxygenase (IDO) is associated with immunological and neurological disorders, and inhibition of its enzyme activity could be a therapeutic approach for treatment of these disorders. The aim of the present study was to establish a large animal model to study the accumulation of the potential IDO inhibitor 1-methyltryptophan (1-MT) in blood and different organs of domestic pigs (Sus scrofa domestica). Because 1-MT has not been previously evaluated in pigs, the pharmacokinetics of a single subcutaneous 1-MT application was investigated. Based on this kinetic study, a profile for repeated 1-MT applications over a period of five days was simulated and tested. The results show that a single administration of 1-MT increases its concentrations in blood, with the maximum concentration being obtained at 12 h. Repeated daily injections of 1‑MT generated increasing plasma concentrations followed by a steady-state after two days. Twelve hours after the final application, accumulation of 1-MT was observed in the brain and other organs, with a substantial variability among various tissues. The concentrations of 1-MT measured in plasma and tissues were similar to, or even higher, than those of tryptophan. Our data indicate that repeated subcutaneous injections of 1-MT provide a suitable model for accumulation of 1-MT in plasma and tissues of domestic pigs. These findings provide a basis for further research on the immunoregulatory functions of IDO in a large animal model.
- Published
- 2015
19. Association between waist circumference and gray matter volume in 2344 individuals from two adult community-based samples
- Author
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Harald J. Freyberger, Jan Terock, Grazyna Domanska, Katrin Hegenscheid, Nele Friedrich, Mohamad Habes, Deborah Janowitz, Hans Jörgen Grabe, Matthias Nauck, Katharina Wittfeld, Norbert Hosten, and Henry Völzke
- Subjects
Adult ,Male ,Waist ,pathology [Obesity] ,Cognitive Neuroscience ,Olfactory sulcus ,Cuneus ,Lingual gyrus ,Sex Factors ,Supramarginal gyrus ,Gyrus ,pathology [Brain] ,pathology [Gray Matter] ,medicine ,Humans ,Obesity ,ddc:610 ,Gray Matter ,Postcentral gyrus ,Age Factors ,Brain ,Anatomy ,Middle Aged ,Magnetic Resonance Imaging ,medicine.anatomical_structure ,Cross-Sectional Studies ,nervous system ,Neurology ,Frontal lobe ,Female ,Waist Circumference ,Psychology ,Neuroscience - Abstract
We analyzed the putative association between abdominal obesity (measured in waist circumference) and gray matter volume (Study of Health in Pomerania: SHIP-2, N=758) adjusted for age and gender by applying volumetric analysis and voxel-based morphometry (VBM) with VBM8 to brain magnetic resonance (MR) imaging. We sought replication in a second, independent population sample (SHIP-TREND, N=1586). In a combined analysis (SHIP-2 and SHIP-TREND) we investigated the impact of hypertension, type II diabetes and blood lipids on the association between waist circumference and gray matter. Volumetric analysis revealed a significant inverse association between waist circumference and gray matter volume. VBM in SHIP-2 indicated distinct inverse associations in the following structures for both hemispheres: frontal lobe, temporal lobes, pre- and postcentral gyrus, supplementary motor area, supramarginal gyrus, insula, cingulate gyrus, caudate nucleus, olfactory sulcus, para-/hippocampus, gyrus rectus, amygdala, globus pallidus, putamen, cerebellum, fusiform and lingual gyrus, (pre-) cuneus and thalamus. These areas were replicated in SHIP-TREND. More than 76% of the voxels with significant gray matter volume reduction in SHIP-2 were also distinct in TREND. These brain areas are involved in cognition, attention to interoceptive signals as satiety or reward and control food intake. Due to our cross-sectional design we cannot clarify the causal direction of the association. However, previous studies described an association between subjects with higher waist circumference and future cognitive decline suggesting a progressive brain alteration in obese subjects. Pathomechanisms may involve chronic inflammation, increased oxidative stress or cellular autophagy associated with obesity.
- Published
- 2015
20. Activation of indoleamine 2,3-dioxygenase by LPS in a porcine model
- Author
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Grazyna Domanska, Winfried Otten, Armin Tuchscherer, Margret Tuchscherer, Ellen Kanitz, K. Wollenhaupt, and Elisa Wirthgen
- Subjects
Lipopolysaccharides ,Male ,medicine.medical_specialty ,Kynurenine pathway ,Swine ,Immunology ,Stimulation ,Biology ,Pharmacology ,Microbiology ,chemistry.chemical_compound ,Internal medicine ,medicine ,Animals ,Humans ,Indoleamine-Pyrrole 2,3,-Dioxygenase ,Indoleamine 2,3-dioxygenase ,Molecular Biology ,Lung ,Cells, Cultured ,Kynurenine ,chemistry.chemical_classification ,Blood Cells ,Tumor Necrosis Factor-alpha ,Tryptophan ,Cell Biology ,Interleukin-10 ,Up-Regulation ,Enzyme Activation ,Infectious Diseases ,Enzyme ,Endocrinology ,chemistry ,Immune System Diseases ,Liver ,Models, Animal ,Feasibility Studies ,Tumor necrosis factor alpha ,Inflammation Mediators ,Ex vivo - Abstract
Indoleamine 2,3-dioxygenase (IDO) is a rate-limiting enzyme for the degradation of tryptophan (Trp) along the kynurenine (Kyn) pathway, and its increased activation is associated with immunologic disorders. Because the specific role of IDO activation is not yet completely clear, the aim of the present study was to establish a pig model of IDO activation for further research. The activation of IDO in pigs was induced experimentally by LPS stimulation in vivo and ex vivo. IDO activation was characterized by measuring Trp, Trp metabolites and IDO protein expression in blood, liver, lung, muscle and different brain areas. The results show that the in vivo LPS administration induced increased plasma concentrations of TNF-α and IL-10, a depletion of Trp and an increase of Kyn, indicating an elevated enzymatic activity of IDO. This was supported by an LPS-induced IDO protein expression in blood, liver and lung. The ex vivo LPS stimulation also resulted in increased TNF-α concentrations and an IDO activation, characterized by an increase of Trp metabolites and IDO protein expression. In conclusion, our data emphasize that the LPS stimulation is a suitable model for IDO activation in the domestic pig, which provides a basis for further research on immunoregulatory IDO functions.
- Published
- 2013
21. Systemic changes of tryptophan catabolites via the indoleamine-2,3-dioxygenase pathway in primary cervical cancer
- Author
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Christina, Fotopoulou, Jalid, Sehouli, Rene, Pschowski, Stephan, VON Haehling, Grazyna, Domanska, Elena-Ioana, Braicu, Gerhard, Fusch, Petra, Reinke, and Joerg C, Schefold
- Subjects
Adult ,Tryptophan ,Humans ,Indoleamine-Pyrrole 2,3,-Dioxygenase ,Uterine Cervical Neoplasms ,Female ,Middle Aged ,Aged - Abstract
Induction of tryptophan catabolism is mediated by inflammatory mechanisms including up-regulation of the immunoregulatory enzyme indoleamine-2,3-dioxygenase (IDO). This leads to the formation of mediators collectively referred to as kynurenines. Kynurenines are involved in various diseases such as renal failure, sepsis and cancer. We aimed to investigate whether systemic levels of kynurenines are induced in primary cervical cancer (PCC).Tryptophan, serotonin, kynurenine, kynurenic acid, quinolinic acid and estimated IDO activity were determined using tandem mass spectrometry for serum samples of 20 PCC patients (mean age: 45.1±11.3 years, FIGO-stage: 1b1-2b) prior to radical abdominal surgery. Data were compared to those from 40 healthy controls. Receiver operating curve (ROC) analyses were performed.Mean tryptophan (22.7±15.1 vs. 18.9±3.5 μM; p=0.27) and kynurenine levels (2.25±0.7 vs. 2.59±0.25 μM; p=0.1) were unchanged in PCC patients when compared to controls. Estimated IDO activity (kynurenine level × 100/tryptophan: 11.8±4.5 vs. 14.1±2.4; p=0.04) and mean levels of kynurenic acid (0.25±0.06 vs. 0.55±0.23 μM; p0.0001) were significantly lower in PCC patients compared to controls, while mean levels of quinolinic acid (0.35±0.07 vs. 0.24±0.09 μM, p0.0001) were significantly higher. The ratio of quinolinic acid to kynurenic acid (Q/K) differed significantly between patients with and those without cancer (p0.0001). When this index was0.95, the sensitivity and specificity for identification of PCC patients were 100% and 90%, respectively (AUC=0.981, 95% CI=0.907-0.999; positive likelihood ratio +10.0).PCC is associated with increased systemic levels of quinolinic acid and reduced levels of kynurenic acid. In our study population, the Q/K allowed identification of PCC patients with a high level of accuracy. The prognostic power and relevance of this novel proposed index remains to be elucidated in further larger prospective studies.
- Published
- 2011
22. Excessive tryptophan catabolism along the kynurenine pathway precedes ongoing sepsis in critically ill patients
- Author
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Grazyna Domanska, B Holtfreter, Petra Reinke, Gerhard Fusch, Christine Schuett, M Gruendling, A Westerholt, Joerg C. Schefold, J P Haas, and Jan-Philip Zeden
- Subjects
Adult ,Male ,medicine.medical_specialty ,Kynurenine pathway ,Critical Illness ,Critical Care and Intensive Care Medicine ,Gastroenterology ,law.invention ,Sepsis ,chemistry.chemical_compound ,law ,Internal medicine ,medicine ,Humans ,Renal Insufficiency ,Intensive care medicine ,Indoleamine 2,3-dioxygenase ,Kynurenine ,Septic shock ,business.industry ,Area under the curve ,Tryptophan ,Middle Aged ,medicine.disease ,NAD ,Intensive care unit ,Anesthesiology and Pain Medicine ,chemistry ,Female ,business ,Quinolinic acid - Abstract
It has recently been shown that an increased plasma level of the tryptophan catabolite kynurenine is an early indicator for the development of sepsis in major trauma patients. We examined the predictive value of kynurenine pathway activity for ongoing sepsis in patients being admitted to a surgical intensive care unit for different reasons. In addition, we asked whether an accumulation of kynurenines in patients’ plasma depends on reduced renal clearance. We conducted a prospective observational study including 100 consecutive patients and monitored laboratory variables, physiological and adverse events, sepsis and outcome. Using tandem mass spectrometry, we quantified the five indoleamines tryptophan, serotonin (5-HT), kynurenine, quinolinic acid and kynurenic acid at baseline and twice a week during the intensive care unit stay. Among the patients enrolled, 50 did not develop sepsis in the intensive care unit (non-septic), 18 patients did not have sepsis at baseline but developed sepsis later on (preseptic) and 32 patients already fulfilled the criteria of severe sepsis and septic shock at baseline (septic). In general, non-septic critically ill patients showed activation of the kynurenine pathway, but septic shock coincided with an exacerbation of kynurenine pathway activity even in the absence of renal failure. Importantly, plasma concentrations of quinolinic acid (area under the curve 0.832 [95% confidence interval 0.710 to 0.954]) and the Quin/Trp ratio (area under the curve 0.835 [95% confidence interval; 0.719 to 0.952]) showed the best discrimination between non-septic and pre-septic patients at baseline. These findings open new avenues for further investigations on the pathophysiology of sepsis.
- Published
- 2010
23. Side effects of control treatment can conceal experimental data when studying stress responses to injection and psychological stress in mice
- Author
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Christine Schuett, Solveig Drude, Jakob Olfe, Annett Geissler, Cornelia Kiank-Nussbaum, Astrid Starke, and Grazyna Domanska
- Subjects
Male ,medicine.medical_specialty ,medicine.medical_treatment ,Intraperitoneal injection ,Sodium Chloride ,medicine.disease_cause ,Handling, Psychological ,Dexamethasone ,Injections ,Mice ,Receptors, Glucocorticoid ,Internal medicine ,Lymphopenia ,medicine ,Psychological stress ,Animals ,Receptor ,Saline ,Glucocorticoids ,Control treatment ,Cyclodextrins ,Mice, Inbred BALB C ,General Veterinary ,business.industry ,Stressor ,medicine.disease ,Mifepristone ,Endocrinology ,Animal Science and Zoology ,Female ,Lymphocytopenia ,Pharmaceutical Vehicles ,business ,Corticosterone ,Glucocorticoid ,Stress, Psychological ,medicine.drug - Abstract
Routine laboratory procedures, such as handling or transporting animals or carrying out injections on animals, are stressful for animals but are necessary in many pre-clinical studies. Here, the authors show that multiple injections of the non-toxic vehicle cyclodextrin moderately increased plasma corticosterone concentrations in female BALB/c mice. Additionally, male BALB/c mice that had received a single intraperitoneal injection of harmless saline had an increased glucocorticoid response to a second saline injection. The authors found that female mice that had been exposed to an acute psychological stress session had a decreased glucocorticoid response to a second homotypic stressor. In contrast, multiple psychological stress sessions led to increased glucocorticoid release in female mice. Acute injection(s) of saline in male mice and of cyclodextrin in female mice led to transient lymphocytopenia. Further analysis showed that repeated stress-induced lymphocytopenia is glucocorticoid-dependent. The authors conclude that laboratory stress can affect physiological parameters in mice, potentially altering study results.
- Published
- 2010
24. Different stress-related phenotypes of BALB/c mice from in-house or vendor: alterations of the sympathetic and HPA axis responsiveness
- Author
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Christine Schuett, Cornelia Kiank, Grazyna Domanska, and Jakob Olfe
- Subjects
medicine.medical_specialty ,Hypothalamo-Hypophyseal System ,Time Factors ,Physiology ,Pituitary-Adrenal System ,Breeding ,lcsh:Physiology ,BALB/c ,chemistry.chemical_compound ,Mice ,Norepinephrine ,Adrenocorticotropic Hormone ,Corticosterone ,Stress, Physiological ,Physiology (medical) ,Internal medicine ,Monoaminergic ,Research article ,medicine ,Animals ,Analysis of Variance ,Mice, Inbred BALB C ,biology ,lcsh:QP1-981 ,Body Weight ,General Medicine ,Reserpine ,biology.organism_classification ,Phenotype ,Housing, Animal ,Monoamine neurotransmitter ,Endocrinology ,chemistry ,Female ,Analysis of variance ,Glucocorticoid ,Stress, Psychological ,medicine.drug - Abstract
Background Laboratory routine procedures such as handling, injection, gavage or transportation are stressful events which may influence physiological parameters of laboratory animals and may interfere with the interpretation of the experimental results. Here, we investigated if female BALB/c mice derived from in-house breeding and BALB/c mice from a vendor which were shipped during their juvenile life differ in their HPA axis activity and stress responsiveness in adulthood. Results We show that already transferring the home cage to another room is a stressful event which causes an increased HPA axis activation for at least 24 hours as well as a loss of circulating lymphocytes which normalizes during a few days after transportation. However and important for the interpretation of experimental data, commercially available strain-, age- and gender-matched animals that were shipped over-night showed elevated glucocorticoid levels for up to three weeks after shipment, indicating a heightened HPA axis activation and they gained less body weight during adolescence. Four weeks after shipment, these vendor-derived mice showed increased corticosterone levels at 45-min after intraperitoneal ACTH challenge but, unexpectedly, no acute stress-induced glucocorticoid release. Surprisingly, activation of monoaminergic pathways were identified to inhibit the central nervous HPA axis activation in the vendor-derived, shipped animals since depletion of monoamines by reserpine treatment could restore the stress-induced HPA axis response during acute stress. Conclusions In-house bred and vendor-derived BALB/c mice show a different stress-induced HPA axis response in adulthood which seems to be associated with different central monoaminergic pathway activity. The stress of shipment itself and/or differences in raising conditions, therefore, can cause the development of different stress response phenotypes which needs to be taken into account when interpreting experimental data.
- Published
- 2010
25. Hypermetabolic syndrome as a consequence of repeated psychological stress in mice
- Author
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Robert Geffers, Cornelia Kiank, Christine Schuett, Uwe Völker, Grazyna Domanska, Gerhard Fusch, Maren Depke, and Ernst-Moritz-Arndt-University, Interfaculty Institute of Genetics and Functional Genomics, 17487 Greifswald, Germany.
- Subjects
medicine.medical_specialty ,Ratón ,Drinking Behavior ,Biology ,Polymerase Chain Reaction ,Mice ,Endocrinology ,Insulin resistance ,In vivo ,Internal medicine ,medicine ,Animals ,Acidosis ,Oligonucleotide Array Sequence Analysis ,Metabolic Syndrome ,Mice, Inbred BALB C ,Metabolic Syndrome X ,Stressor ,Water ,medicine.disease ,Disease Models, Animal ,Kinetics ,Gluconeogenesis ,Acoustic Stimulation ,Acute Disease ,Lean body mass ,Female ,medicine.symptom ,Energy Intake ,Energy Metabolism ,Dyslipidemia ,Stress, Psychological - Abstract
Stress is a powerful modulator of neuroendocrine, behavioral, and immunological functions. After 4.5-d repeated combined acoustic and restraint stress as a murine model of chronic psychological stress, severe metabolic dysregulations became detectable in female BALB/c mice. Stress-induced alterations of metabolic processes that were found in a hepatic mRNA expression profiling were verified by in vivo analyses. Repeatedly stressed mice developed a hypermetabolic syndrome with the severe loss of lean body mass, hyperglycemia, dyslipidemia, increased amino acid turnover, and acidosis. This was associated with hypercortisolism, hyperleptinemia, insulin resistance, and hypothyroidism. In contrast, after a single acute stress exposure, changes in expression of metabolic genes were much less pronounced and predominantly confined to gluconeogenesis, probably indicating that metabolic disturbances might be initiated already early but will only manifest in repeatedly stressed mice. Thus, in our murine model, repeated stress caused severe metabolic dysregulations, leading to a drastic reduction of the individual’s energy reserves. Under such circumstances stress may further reduce the ability to cope with new stressors such as infection or cancer.
- Published
- 2008
26. Protein targeting to mitochondria of Saccharomyces cerevisiae and Neurospora crassa: in vitro and in vivo studies
- Author
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Panagiotis, Papatheodorou, Grazyna, Domanska, and Joachim, Rassow
- Subjects
Protein Transport ,Neurospora crassa ,Green Fluorescent Proteins ,Saccharomyces cerevisiae ,In Vitro Techniques ,Protein Precursors ,Biochemistry ,Mitochondria - Abstract
Most studies on the biogenesis of mitochondrial proteins have been carried out using fungal mitochondria as a model system. In particular, baker's yeast, Saccharomyces cerevisiae, combines several experimental advantages, allowing both genetic and biochemical approaches and thus a combination of investigations in vivo and in vitro. However, the red bread mold Neurospora crassa has also been an important research tool. Isolated mitochondria can be used from both organisms for import experiments in a reconstituted system, using radiolabeled precursor proteins synthesized in reticulocyte lysate or purified preproteins. Assays are available for studies on the import pathways and localization of mitochondrial proteins and for the characterization of the components of the protein import machinery.
- Published
- 2007
27. Protein Targeting to Mitochondria of Saccharomyces cerevisiae and Neurospora crassa: In Vitro and In Vivo Studies
- Author
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Panagiotis Papatheodorou, Grazyna Domanska, and Joachim Rassow
- Published
- 2007
28. Protein Targeting to Mitochondria of Saccharomyces cerevisiae and Neurospora crassa
- Author
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Grazyna Domanska, Panagiotis Papatheodorou, and Joachim Rassow
- Subjects
Biochemistry ,biology ,Protein targeting ,Saccharomyces cerevisiae ,medicine ,Mitochondrion ,biology.organism_classification ,medicine.disease_cause ,In vitro ,Yeast ,Biogenesis ,Neurospora crassa ,Transport protein - Abstract
Most studies on the biogenesis of mitochondrial proteins have been carried out using fungal mitochondria as a model system. In particular, baker's yeast, Saccharomyces cerevisiae, combines several experimental advantages, allowing both genetic and biochemical approaches and thus a combination of investigations in vivo and in vitro. However, the red bread mold Neurospora crassa has also been an important research tool. Isolated mitochondria can be used from both organisms for import experiments in a reconstituted system, using radiolabeled precursor proteins synthesized in reticulocyte lysate or purified preproteins. Assays are available for studies on the import pathways and localization of mitochondrial proteins and for the characterization of the components of the protein import machinery.
- Published
- 2007
29. The enteropathogenic Escherichia coli (EPEC) Map effector is imported into the mitochondrial matrix by the TOM/Hsp70 system and alters organelle morphology
- Author
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Brendan Kenny, Olaf Selchow, Grazyna Domanska, Panagiotis Papatheodorou, Johannes Mathieu, Joachim Rassow, and Marius Oxle
- Subjects
Saccharomyces cerevisiae Proteins ,Immunology ,Amino Acid Motifs ,Molecular Sequence Data ,Apoptosis ,Saccharomyces cerevisiae ,Mitochondrion ,In Vitro Techniques ,Microbiology ,Mitochondrial Membrane Transport Proteins ,Membrane Potentials ,Mitochondrial membrane transport protein ,Virology ,Escherichia coli ,Translocase ,Animals ,HSP70 Heat-Shock Proteins ,Amino Acid Sequence ,biology ,Effector ,Escherichia coli Proteins ,Intracellular Signaling Peptides and Proteins ,Membrane Transport Proteins ,Transport protein ,Cell biology ,Mitochondria ,Rats ,Protein Transport ,Biochemistry ,Cytoplasm ,Chaperone (protein) ,Mitochondrial Membranes ,biology.protein ,Rabbits ,Bacterial outer membrane ,Carrier Proteins - Abstract
Enteropathogenic Escherichia coli (EPEC) is a human intestinal pathogen and a major cause of diarrhoea, particularly among infants in developing countries. EPEC target the Map and EspF multifunctional effector proteins to host mitochondria - organelles that play crucial roles in regulating cellular processes such as programmed cell death (apoptosis). While both molecules interfere with the organelles ability to maintain a membrane potential, EspF plays the predominant role and is responsible for triggering cell death. To learn more about the Map-mitochondria interaction, we studied Map localization to mitochondria with purified mitochondria (from mammalian and yeast cells) and within intact yeast. This revealed that (i) Map targeting is dependent on the predicted N-terminal mitochondrial targeting sequence, (ii) the N-terminal 44 residues are sufficient to target proteins to mitochondria and (iii) Map import involves the mitochondrial outer membrane translocase (Tom22 and Tom40), the mitochondrial membrane potential, and the matrix chaperone, mtHsp70. These results are consistent with Map import into the mitochondria matrix via the classical import mechanism. As all known, Map-associated phenotypes in mammalian cells are independent of mitochondrial targeting, this may indicate that import serves as a mechanism to remove Map from the cytoplasm thereby regulating cytoplasmic function. Intriguingly, Map, but not EspF, alters mitochondrial morphology with deletion analysis revealing important roles for residues 101-152. Changes in mitochondrial morphology have been linked to alterations in the ability of these organelles to regulate cellular processes providing a possible additional role for Map import into mitochondria.
- Published
- 2006
30. Conserved mechanism of Oxa1 insertion into the mitochondrial inner membrane
- Author
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Grazyna Domanska, Elke A Dian, Christian Motz, Joachim Rassow, Olga Randelj, Thomas Krimmer, and Sebastian Reif
- Subjects
Saccharomyces cerevisiae Proteins ,Translocase of the outer membrane ,Saccharomyces cerevisiae ,Biology ,Electron Transport Complex IV ,Mitochondrial Proteins ,Mitochondrial membrane transport protein ,Structural Biology ,Mitochondrial Precursor Protein Import Complex Proteins ,HSP70 Heat-Shock Proteins ,Molecular Biology ,Integral membrane protein ,Peripheral membrane protein ,Membrane Transport Proteins ,Nuclear Proteins ,Intracellular Membranes ,Cell biology ,Mitochondria ,Molecular Weight ,Membrane protein ,Translocase of the inner membrane ,Mutation ,biology.protein ,ATP–ADP translocase ,Intermembrane space ,Protein Binding - Abstract
Oxa1 is the mitochondrial representative of a family of related proteins that mediate the insertion of substrate proteins into the membranes of bacteria, chloroplasts, and mitochondria. Several studies have demonstrated that the bacterial homologue YidC participates both in the direct uptake of proteins from the bacterial cytosol, and in the uptake of nascent proteins from the Sec translocase. Studies on the biogenesis of membrane proteins in mitochondria established that Oxa1 has the capability to receive substrates at the inner surface of the inner membrane. In this study, we asked if Oxa1 may similarly cooperate with a protein translocase within the membrane. Since Oxa1 is involved in its own biogenesis, we used the precursor of Oxa1 as a model protein and investigated its import pathway. We found that immediately after import into mitochondria, Oxa1 initially accumulates at Tim23 that forms the inner membrane protein translocase. Cleavage of the Oxa1 presequence is dependent on mtHsp70, a heat shock protein of the mitochondrial matrix. However, mutant mtHsp70 showing a defect in the release of bound substrate proteins does not interfere with subsequent membrane insertion, indicating that membrane insertion of the mature protein is essentially mtHsp70-independent. We conclude that Oxa1 has the ability to accept preproteins within the membrane.
- Published
- 2005
31. 32. Modulation of HPA-axis responsiveness by laboratory stress or shipment of mice alters experimental data
- Author
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S. Drude, J. Olfe, Christine Schuett, Grazyna Domanska, A. Geissler, and Cornelia Kiank
- Subjects
Stress (mechanics) ,Behavioral Neuroscience ,Endocrine and Autonomic Systems ,Modulation ,Chemistry ,Immunology ,Experimental data ,Cell biology - Published
- 2009
32. 65. Proinflammatory cytokines mediate acute stress-induced loss of intestinal barrier function in the terminal ileum which enhances tryptophan catabolism in BALB/C mice
- Author
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Cornelia Kiank, Christine Schuett, S. Voss, Jan-Philip Zeden, Grazyna Domanska, Astrid Starke, Yvette Taché, and Gerhard Fusch
- Subjects
medicine.medical_specialty ,biology ,Endocrine and Autonomic Systems ,Chemistry ,Immunology ,biology.organism_classification ,BALB/c ,Tryptophan catabolism ,Proinflammatory cytokine ,Behavioral Neuroscience ,Endocrinology ,medicine.anatomical_structure ,Internal medicine ,medicine ,Terminal ileum ,Acute stress ,Barrier function - Published
- 2009
33. 69. Hypermetabolic syndrome as a consequence of chronic psychological stress in mice
- Author
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Robert Geffers, Gerhard Fusch, Grazyna Domanska, Maren Depke, Cornelia Kiank, Christine Schuett, and Uwe Voelker
- Subjects
Behavioral Neuroscience ,Endocrine and Autonomic Systems ,business.industry ,Immunology ,Physiology ,Medicine ,Psychological stress ,business ,medicine.disease_cause - Published
- 2008
34. Stress-like CRF1 receptor signaling: Anti-inflammatory immune conditioning and altered tight junction protein expression in the proximal colon of female BALB/c mice
- Author
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Yvette Taché, Grazyna Domanska, Mulugeta Million, Muriel H. Larauche, Pu-Qing Yuan, and Cornelia Kiank
- Subjects
biology ,Tight junction ,Endocrine and Autonomic Systems ,medicine.drug_class ,Chemistry ,Immunology ,Crf1 receptor ,biology.organism_classification ,Protein expression ,Anti-inflammatory ,BALB/c ,Cell biology ,Behavioral Neuroscience ,Immune system ,medicine ,Proximal colon - Published
- 2010
35. Helicobacter pylori VacA Toxin/Subunit p34: Targeting of an Anion Channel to the Inner Mitochondrial Membrane
- Author
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Elke A. Dian-Lothrop, Joachim Rassow, Antoine Galmiche, Oliver Kepp, Kathrin Günnewig, Grazyna Domanska, Anke Harsman, Christian Motz, Richard Wagner, Lars Becker, Michael Meinecke, Boris Reljic, and Panagiotis Papatheodorou
- Subjects
QH301-705.5 ,Protein subunit ,Immunology ,TIM/TOM complex ,Mitochondrion ,Biology ,Microbiology ,Mitochondrial apoptosis-induced channel ,Cell Biology/Membranes and Sorting ,Bacterial Proteins ,Virology ,Genetics ,Animals ,Humans ,Biology (General) ,Inner mitochondrial membrane ,Molecular Biology ,Helicobacter pylori ,RC581-607 ,Rats ,Electrophysiology ,Microscopy, Fluorescence ,Membrane protein ,Biochemistry ,Biophysics/Membrane Proteins and Energy Transduction ,Mitochondrial Membranes ,Translocase of the inner membrane ,Parasitology ,Immunologic diseases. Allergy ,Microbiology/Cellular Microbiology and Pathogenesis ,Bacterial outer membrane ,Research Article ,HeLa Cells - Abstract
The vacuolating toxin VacA, released by Helicobacter pylori, is an important virulence factor in the pathogenesis of gastritis and gastroduodenal ulcers. VacA contains two subunits: The p58 subunit mediates entry into target cells, and the p34 subunit mediates targeting to mitochondria and is essential for toxicity. In this study we found that targeting to mitochondria is dependent on a unique signal sequence of 32 uncharged amino acid residues at the p34 N-terminus. Mitochondrial import of p34 is mediated by the import receptor Tom20 and the import channel of the outer membrane TOM complex, leading to insertion of p34 into the mitochondrial inner membrane. p34 assembles in homo-hexamers of extraordinary high stability. CD spectra of the purified protein indicate a content of >40% β-strands, similar to pore-forming β-barrel proteins. p34 forms an anion channel with a conductivity of about 12 pS in 1.5 M KCl buffer. Oligomerization and channel formation are independent both of the 32 uncharged N-terminal residues and of the p58 subunit of the toxin. The conductivity is efficiently blocked by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), a reagent known to inhibit VacA-mediated apoptosis. We conclude that p34 essentially acts as a small pore-forming toxin, targeted to the mitochondrial inner membrane by a special hydrophobic N-terminal signal., Author Summary VacA is a toxic protein produced by Helicobacter pylori, the bacteria that cause gastritis and ulcer diseases. p34, the toxic component of VacA, is known to damage mitochondria, defined cell organelles in the target cells. However, both the mechanism of mitochondrial targeting and the toxic activity inside the mitochondria are unclear. In this study, we show that p34 carries a unique targeting signal that is different from all targeting signatures that were previously identified in endogenous mitochondrial proteins. Eventually, p34 seems to act as an anion channel in the mitochondrial inner membrane and thus to destroy the balance of salt ions in the organelles.
- Published
- 2010
36. 91. Chronic psychological stress in mice: Interference of impulsive overreaction with depression-like behavioral alterations
- Author
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Cornelia Kiank, Christine Schuett, Grazyna Domanska, D. Muschter, and Winfried Otten
- Subjects
Behavioral Neuroscience ,Endocrine and Autonomic Systems ,business.industry ,Immunology ,Medicine ,Psychological stress ,business ,Interference (genetic) ,medicine.disease_cause ,Depression (differential diagnoses) ,Clinical psychology - Published
- 2009
37. Helicobacter pylori VacA toxin/subunit p34: targeting of an anion channel to the inner mitochondrial membrane.
- Author
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Grazyna Domańska, Christian Motz, Michael Meinecke, Anke Harsman, Panagiotis Papatheodorou, Boris Reljic, Elke A Dian-Lothrop, Antoine Galmiche, Oliver Kepp, Lars Becker, Kathrin Günnewig, Richard Wagner, and Joachim Rassow
- Subjects
Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The vacuolating toxin VacA, released by Helicobacter pylori, is an important virulence factor in the pathogenesis of gastritis and gastroduodenal ulcers. VacA contains two subunits: The p58 subunit mediates entry into target cells, and the p34 subunit mediates targeting to mitochondria and is essential for toxicity. In this study we found that targeting to mitochondria is dependent on a unique signal sequence of 32 uncharged amino acid residues at the p34 N-terminus. Mitochondrial import of p34 is mediated by the import receptor Tom20 and the import channel of the outer membrane TOM complex, leading to insertion of p34 into the mitochondrial inner membrane. p34 assembles in homo-hexamers of extraordinary high stability. CD spectra of the purified protein indicate a content of >40% beta-strands, similar to pore-forming beta-barrel proteins. p34 forms an anion channel with a conductivity of about 12 pS in 1.5 M KCl buffer. Oligomerization and channel formation are independent both of the 32 uncharged N-terminal residues and of the p58 subunit of the toxin. The conductivity is efficiently blocked by 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), a reagent known to inhibit VacA-mediated apoptosis. We conclude that p34 essentially acts as a small pore-forming toxin, targeted to the mitochondrial inner membrane by a special hydrophobic N-terminal signal.
- Published
- 2010
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