Ingo Nolte

Publications (107)222 Total impact

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  Available from: Siegfried Wagner

  Article: Comparison of non-coding RNAs in human and canine cancer

  Siegfried Wagner, Saskia Willenbrock, Ingo Nolte, Hugo Murua Escobar
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  ABSTRACT: Thediscoveryofthepost-transcriptionalgenesilencing(PTGS)bysmallnon-protein-codingRNAsisconsideredasamajorbreakthroughinbiology.Inthelastdecadewejuststartedtorealizethebiologicfunctionandcomplexityofgeneregulationbysmallnon-codingRNAs.PTGSisaconservedphenomenonwhichwasobservedinvariousspeciessuchasfungi,worms,plants,andmammals.MicroRNAs(miRNA)andsmallinterferingRNAs(siRNAs)aretwogenesilencingmediatorsconstitutinganevolutionaryconservedclassofnon-codingRNAsregulatingmanybiologicalprocessesineukaryotes.AsthissmallRNAsappeartoregulategeneexpressionattranslationalandtranscriptionallevelitisnotsurprisingthatduringthelastdecademanyhumandiseasesamongthemAlzheimer’sdisease,cardiovasculardiseases,andvariouscancertypeswereassociatedwithderegulatedmiRNAexpression.ConsequentlysmallRNAsareconsideredtoholdbigpromisesastherapeuticagents.However,despiteoftheenormoustherapeuticpotentialmanyquestionsremainunanswered.Amajorcriticalpoint,whenevaluatingnoveltherapeuticapproaches,isthetransferofinvitrosettingstoaninvivomodel.Classicalanimalmodelsrelyonthelaboratorykeptanimalsunderartificialconditionsandoftenmissinganintactimmunesystem.Modelorganismswithspontaneouslyoccurringtumorsase.g.,dogsprovidethepossibilitytoevaluatetherapeuticagentsunderthesurveillanceofaninintactimmunesystemandtherebyprovidinganauthentictumorreactingscenario.ConsideringthegenomicsimilaritybetweencaninesandhumansandtheadvantagesofthedogascancermodelsystemforhumanneoplasiastheanalysesofthecomplexroleofsmallRNAsincaninetumordevelopmentcouldbeofmajorvalueforbothspecies.HereinwediscusscomparativelytheroleofmiRNAsinhumanandcaninecancerdevelopmentandhighlightthepotentialandadvantagesofthemodelorganismdogfortumorresearch.
  Frontiers in Genetics. 04/2013; 4(46).
• Article: Establishment and Evaluation of a Bead-based Luminex Assay Allowing Simultaneous Quantification of Equine IL-12 and IFN-γ.

  Maria Carolina Duran, Saskia Willenbrock, Jessika-M V Müller, Ingo Nolte, Karsten Feige, Hugo Murua Escobar
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  ABSTRACT: Background/Aim: Interleukin-12 (IL-12) and interferon gamma (IFN-γ) are key cytokines in immunemediated equine melanoma therapy. Currently, a method for accurate simultaneous quantification of these equine cytokines is lacking. Therefore, we sought to establish an assay that allows for accurate and simultaneous quantification of equine IL-12 (eIL-12) and IFN-γ (eIFN-γ). Several antibodies were evaluated for cross-reactivity to eIL-12 and eIFN-γ and were used to establish a bead-based Luminex assay, which was subsequently applied to quantify cytokine concentrations in biological samples. Cytokine detection ranged from 31.5-5,000 pg/ml and 15-10,000 pg/ml for eIL-12 and eIFN-γ, respectively. eIL-12 was detected in supernatants of stimulated peripheral blood mononuclear cells (PBMCs) and supernatants/cell lysates of eIL-12 expression plasmid-transfected cells. Low or undetectable cytokine concentrations were measured in negative controls. In equine serum samples, the mean measured eIL-12 concentration was 1,374±8 pg/ml. The bead-based assay and ELISA for eIFN-γ used to measure eIFN-γ concentrations, showed similar concentrations. Results demonstrate, to our knowledge for the first time, that cross-reactive antibody pairs to eIL-12 and eIFN-γ and Luminex bead-based technology allow for accurate, simultaneous and multiplexed quantification of these key cytokines in biological samples.
  Anticancer research 04/2013; 33(4):1325-36. · 1.73 Impact Factor
• Article: Comparison of non-coding RNAs in human and canine cancer.

  Siegfried Wagner, Saskia Willenbrock, Ingo Nolte, Hugo Murua Escobar
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  ABSTRACT: The discovery of the post-transcriptional gene silencing (PTGS) by small non-protein-coding RNAs is considered as a major breakthrough in biology. In the last decade we just started to realize the biologic function and complexity of gene regulation by small non-coding RNAs. PTGS is a conserved phenomenon which was observed in various species such as fungi, worms, plants, and mammals. Micro RNAs (miRNA) and small interfering RNAs (siRNAs) are two gene silencing mediators constituting an evolutionary conserved class of non-coding RNAs regulating many biological processes in eukaryotes. As this small RNAs appear to regulate gene expression at translational and transcriptional level it is not surprising that during the last decade many human diseases among them Alzheimer's disease, cardiovascular diseases, and various cancer types were associated with deregulated miRNA expression. Consequently small RNAs are considered to hold big promises as therapeutic agents. However, despite of the enormous therapeutic potential many questions remain unanswered. A major critical point, when evaluating novel therapeutic approaches, is the transfer of in vitro settings to an in vivo model. Classical animal models rely on the laboratory kept animals under artificial conditions and often missing an intact immune system. Model organisms with spontaneously occurring tumors as e.g., dogs provide the possibility to evaluate therapeutic agents under the surveillance of an in intact immune system and thereby providing an authentic tumor reacting scenario. Considering the genomic similarity between canines and humans and the advantages of the dog as cancer model system for human neoplasias the analyses of the complex role of small RNAs in canine tumor development could be of major value for both species. Herein we discuss comparatively the role of miRNAs in human and canine cancer development and highlight the potential and advantages of the model organism dog for tumor research.
  Frontiers in genetics. 01/2013; 4:46.
• Article: RAGE Splicing Variants in Mammals.

  Katharina Anna Sterenczak, Ingo Nolte, Hugo Murua Escobar
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  ABSTRACT: The receptor for advanced glycation end products (RAGE) is a multiligand receptor of environmental stressors which plays key roles in pathophysiological processes, including immune/inflammatory disorders, Alzheimer's disease, diabetic arteriosclerosis, tumorigenesis, and metastasis. Besides the full-length RAGE protein in humans nearly 20 natural occurring RAGE splicing variants were described on mRNA and protein level. These naturally occurring isoforms are characterized by either N-terminally or C-terminally truncations and are discussed as possible regulators of the full-length RAGE receptor either by competitive ligand binding or by displacing the full-length protein in the membrane. Accordingly, expression deregulations of the naturally occurring isoforms were supposed to have significant effect on RAGE-mediated disorders. Thereby the soluble C-truncated RAGE isoforms present in plasma and tissues are the mostly focused isoforms in research and clinics. Deregulations of the circulating levels of soluble RAGE forms were reported in several RAGE-associated pathological disorders including for example atherosclerosis, diabetes, renal failure, Alzheimer's disease, and several cancer types.Regarding other mammalian species, the canine RAGE gene showed high similarities to the corresponding human structures indicating RAGE to be evolutionary highly conserved between both species. Similar to humans the canine RAGE showed a complex and extensive splicing activity leading to a manifold pattern of RAGE isoforms. Due to the similarities seen in several canine and human diseases-including cancer-comparative structural and functional analyses allow the development of RAGE and ligand-specific therapeutic approaches beneficial for human and veterinary medicine.
  Methods in molecular biology (Clifton, N.J.) 01/2013; 963:265-76.
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  Available from: Iris F. Chaberny

  Dataset: Nienhoff- MRSPseudointermedius among cats admitted to a veterinary teaching hospital

  Ulrike Nienhoff, Kristina Kadlec, Iris F Chaberny, Jutta Verspohl, Gerald-F Gerlach, Stefan Schwarz, Lothar Kreienbrock, Ingo Nolte, Daniela Simon
• Article: Permanent activation of HMGA2 in lipomas mimics its temporal physiological activation linked to the gain of adipose tissue

  Helge W. Thies, Ingo Nolte, Heiner Wenk, Fredrik Mertens, Jörn Bullerdiek, Dominique N. Markowski
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  ABSTRACT: Apparently, up-regulation of HMGA2 in lipomas with 12q14∼15 rearrangement contributes directly to tumorgenesis of these lesions, but the cell of origin has not been identified, yet. In white adipose tissue, new fat cells develop from their progenitors to keep homeostasis or as a result of overnutrition. It is tempting to speculate, that in lipomas as well as a prerequisite of normal gain of adipose tissue stem and/or progenitor cells are targeted by appropiate stimuli to initiate proliferation and differentiation. In this study the activation of HMGA2 by FGF1-driven stimulation of adipose tissue derived stem cells and its overexpression in adipose tissue tumors were analyzed. In addition, the expression of HMGA2 and PPAR-gamma mRNA were quantified in canine subcutaneous abdominal adipose tissue from normal and overweight purebred dogs. The results suggest that the permanent expression of HMGA2 due to chromosomal rearrangements in lipomas mimics its temporal activation resulting from stimuli like FGF1. Furthermore a highly significant increase of HMGA2 expression and a concomitant significant decrease of PPAR-gamma expression was noted in dogs with obesity. These findings suggest the expression of HMGA2 in subcutaneous abdominal adipose tissue as a marker indicating ongoing expansion of fat.
  Obesity 10/2012; · 4.28 Impact Factor
• Article: Longitudinal MRI contrast enhanced monitoring of early tumour development with manganese chloride (MnCl2) and superparamagnetic iron oxide nanoparticles (SPIOs) in a CT1258 based in vivo model of prostate cancer.

  Katharina A Sterenczak, Martin Meier, Silke Glage, Matthias Meyer, Saskia Willenbrock, Patrick Wefstaedt, Martina Dorsch, Jörn Bullerdiek, Hugo Murua Escobar, Hans Hedrich, Ingo Nolte
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  ABSTRACT: Cell lines represent a key tool in cancer research allowing the generation of neoplasias which resemble initial tumours in in-vivo animal models. The characterisation of early tumour development is of major interest in order to evaluate the efficacy of therapeutic agents. Magnetic resonance imaging (MRI) based in-vivo characterisation allows visualisation and characterisation of tumour development in early stages prior to manual palpation. Contrast agents for MRI such as superparamagnetic iron oxide nanoparticles (SPIOs) and manganese chloride (MnCl2) represent powerful tools for the in-vivo characterisation of early stage tumours. In this experimental study, we labelled prostate cancer cells with MnCl2 or SPIOs in vitro and used 1 T MRI for tracing labelled cells in-vitro and 7 T MRI for tracking in an in-vivo animal model. Labelling of prostate cancer cells CT1258 was established in-vitro with MnCl2 and SPIOs. In-vitro detection of labelled cells in an agar phantom was carried out through 1 T MRI while in-vivo detection was performed using 7 T MRI after subcutaneous (s.c.) injection of labelled cells into NOD-Scid mice (n = 20). The animals were scanned in regular intervals until euthanization. The respective tumour volumes were analysed and corresponding tumour masses were subjected to histologic examination. MnCl2in-vitro labelling resulted in no significant metabolic effects on proliferation and cell vitality. In-vitro detection-limit accounted 105 cells for MnCl2 as well as for SPIOs labelling. In-vivo 7 T MRI scans allowed detection of 103 and 104 cells. In-vivo MnCl2 labelled cells were detectable from days 4-16 while SPIO labelling allowed detection until 4 days after s.c. injection. MnCl2 labelled cells were highly tumourigenic in NOD-Scid mice and the tumour volume development was characterised in a time dependent manner. The amount of injected cells correlated with tumour size development and disease progression. Histological analysis of the induced tumour masses demonstrated characteristic morphologies of prostate adenocarcinoma. To the best of our knowledge, this is the first study reporting direct in-vitro MnCl2 labelling and 7 T based in-vivo MRI tracing of cancer cells in a model of prostate cancer. MnCl2 labelling was found to be suitable for in-vivo tracing allowing long detection periods. The labelled cells kept their highly tumourigenic potential in-vivo. Tumour volume development was visualised prior to manual palpation allowing tumour characterisation in early stages of the disease.
  BMC Cancer 07/2012; 12:284. · 3.01 Impact Factor
• Article: Flow cytometric evaluation of peripheral blood and bone marrow and fine-needle aspirate samples from multiple sites in dogs with multicentric lymphoma.

  Alexa E Joetzke, Nina Eberle, Ingo Nolte, Reinhard Mischke, Daniela Simon
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  ABSTRACT: To determine whether the extent of disease in dogs with lymphoma can be assessed via flow cytometry and to evaluate the suitability of fine-needle aspirates from the liver and spleen of dogs for flow cytometric examination. 44 dogs with multicentric B-cell (n = 35) or T-cell lymphoma (9) and 5 healthy control dogs. Procedures-Peripheral blood and bone marrow samples and fine-needle aspirates of lymph node, liver, and spleen were examined via flow cytometry. Logarithmically transformed T-cell-to-B-cell percentage ratio (log[T:B]) values were calculated. Thresholds defined by use of log(T:B) values of samples from control dogs were used to determine extranodal lymphoma involvement in lymphoma-affected dogs; results were compared with cytologic findings. 12 of 245 (5%) samples (9 liver, 1 spleen, and 2 bone marrow) had insufficient cellularity for flow cytometric evaluation. Mean log(T:B) values of samples from dogs with B-cell lymphoma were significantly lower than those of samples from the same site in dogs with T-cell lymphoma and in control dogs. In dogs with T-cell lymphoma, the log(T:B) of lymph node, bone marrow, and spleen samples was significantly higher than in control dogs. Of 165 samples assessed for extranodal lymphoma involvement, 116 (70%) tested positive via flow cytometric analysis; results agreed with cytologic findings in 133 of 161 (83%) samples evaluated via both methods. Results suggested that flow cytometry may aid in detection of extranodal lymphoma involvement in dogs, but further research is needed. Most fine-needle aspirates of liver and spleen were suitable for flow cytometric evaluation.
  American Journal of Veterinary Research 06/2012; 73(6):884-93. · 1.27 Impact Factor
• Article: Bone remodeling after total hip arthroplasty with a short stemmed metaphyseal loading implant: Finite element analysis validated by a prospective DEXA investigation.

  Matthias Lerch, Agnes Kurtz, Christina Stukenborg-Colsman, Ingo Nolte, Nelly Weigel, Anas Bouguecha, Bernd A Behrens
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  ABSTRACT: In total hip arthroplasty (THA), short stemmed cementless implants are used because they are thought to stimulate physiological bone remodeling and reduce stress shielding. We performed a numerical investigation on bone remodeling after implantation of a specific short stemmed implant using finite element analysis (FEA). Overall bone mass loss was 2.8% in the entire femur. Bone mass decrease was mostly found in the proximal part of the calcar and in the greater trochanter due to the vast cross section of the implant, probably leading to stress shielding. In the diaphysis, no change in the apparent bone density was proven. The assumptions made agreed well with bone remodeling data from THA recipients who underwent dual-energy X-ray absorptiometry. However, the clinical investigation revealed a bone mass increase in the minor trochanter region that was less pronounced in the FEA. Further comparisons to other stem designs must be done to verify if the relative advantages of the investigated implant can be accepted. © 2012 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 30:1822-1829, 2012.
  Journal of Orthopaedic Research 04/2012; 30(11):1822-9. · 2.81 Impact Factor
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  Available from: Johannes Junginger

  Article: Immunohistochemical investigation of Foxp3 expression in the intestine in healthy and diseased dogs.

  Johannes Junginger, Ulrike Schwittlick, Frederik Lemensieck, Ingo Nolte, Marion Hewicker-Trautwein
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  ABSTRACT: Intestinal immune regulation including development of oral tolerance is of great importance for the maintenance of intestinal homeostasis. Concerning this, regulatory T cells (Tregs) occupy a pivotal role in cell-mediated immunosuppression. Dysregulation of mucosal immunology leading to an abnormal interaction with commensal bacteria is suggested to play a key role in the pathogenesis of Inflammatory Bowel Disease (IBD) in men and dogs. The aim of this study was to characterise the expression of Foxp3 in the normal canine gut of 18 dogs (mean age: 6.03 years), in 16 dogs suffering from IBD (mean age: 5.05 years), and of 6 dogs with intestinal nematode infection (mean age: 0.87 years) using immunohistochemistry. In the duodenum, Tregs in healthy dogs declined from villi (median: 10.67/62 500 μm2) to crypts (median: 1.89/62 500 μm2). Tregs were further increased in the villi of middle-aged dogs (median: 18.92/62 500 μm2) in contrast to juvenile (median: 3.50/62 500 μm2) and old (median: 9.56/62 500 μm2) individuals. Compared to healthy controls, animals suffering from IBD revealed reduced numbers of Tregs in duodenal villi (median: 4.13/62 500 μm2). Dogs with intestinal nematode infection displayed increased numbers of Tregs (median: 21.06/62 500 μm2) compared to healthy animals.Age-related changes indicate a progressive establishment of oral tolerance and immunosenescence in the canine elderly. The results further suggest that a defect in Treg homeostasis may be involved in the pathogenesis of canine IBD. In contrast, increased numbers of Tregs in the duodenum may be due to nematode infection.
  Veterinary Research 03/2012; 43(1):23. · 4.06 Impact Factor
• Article: Comparison of multi-detector row computed tomography with echocardiography for assessment of left ventricular function in healthy dogs.

  Christiane R Henjes, Stephan Hungerbühler, Iwona B Bojarski, Ingo Nolte, Patrick Wefstaedt
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  ABSTRACT: To evaluate the use of retrospectively ECG-gated, contrast-enhanced, multi-detector row computed tomography (MDCT) for assessment of left ventricular function in dogs and to compare the results with those obtained by use of 2-D and M-mode echocardiographc techniques. 10 healthy Beagles. Dogs underwent MDCT (performed by use of a 64-detector row CT system) and echocardiography under general anesthesia. Left ventricular end-systolic volume (ESV), end-diastolic volume (EDV), and ejection fraction (EF) were determined in MDCT-generated multiplanar reformatted images by use of Simpson and biplane area-length calculation methods. Results were compared with left ventricular ESV, EDV, and EF determined in echocardiographc images by use of Teichholz and bullet method calculations. Results were evaluated via Deming regression analysis and Pearson correlation tests. Bland-Altman analysis was used to assess limits of agreement and systematic errors between the 2 methods. Mean values for EDV and ESV determined by use of MDCT were highly correlated with those determined by use of echocardiography, regardless of the calculation methods compared (r = 0.91 to 0.96); volumes determined by use of MDCT appeared to be higher than those determined by use of echocardiography, although most differences were nonsignificant. Mean EF determined by use of MDCT with the Simpson calculation method was highly correlated with that determined by use of echocardiography with bullet method calculations (r = 0.90). Results suggested that assessment of left ventricular volume and function in dogs is feasible with MDCT. To estimate left ventricular EF with MDCT. use of the Simpson calculation method is advised.
  American Journal of Veterinary Research 03/2012; 73(3):393-403. · 1.27 Impact Factor
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  Available from: Sandeep S Patil

  Article: Oncolytic virotherapy in veterinary medicine: current status and future prospects for canine patients.

  Sandeep S Patil, Ivaylo Gentschev, Ingo Nolte, Gregory Ogilvie, Aladar A Szalay
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  ABSTRACT: Oncolytic viruses refer to those that are able to eliminate malignancies by direct targeting and lysis of cancer cells, leaving non-cancerous tissues unharmed. Several oncolytic viruses including adenovirus strains, canine distemper virus and vaccinia virus strains have been used for canine cancer therapy in preclinical studies. However, in contrast to human studies, clinical trials with oncolytic viruses for canine cancer patients have not been reported. An 'ideal' virus has yet to be identified. This review is focused on the prospective use of oncolytic viruses in the treatment of canine tumors - a knowledge that will undoubtedly contribute to the development of oncolytic viral agents for canine cancer therapy in the future.
  Journal of Translational Medicine 01/2012; 10:3. · 3.41 Impact Factor
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  Available from: Hugo Murua Escobar

  Article: Authentication of primordial characteristics of the CLBL-1 cell line prove the integrity of a canine B-cell lymphoma in a murine in vivo model.

  Barbara C Rütgen, Saskia Willenbrock, Nicola Reimann-Berg, Ingrid Walter, Andrea Fuchs-Baumgartinger, Siegfried Wagner, Boris Kovacic, Sabine E Essler, Ilse Schwendenwein, Ingo Nolte, Armin Saalmüller, Hugo Murua Escobar
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  ABSTRACT: Cell lines are key tools in cancer research allowing the generation of neoplasias in animal models resembling the initial tumours able to mimic the original neoplasias closely in vivo. Canine lymphoma is the major hematopoietic malignancy in dogs and considered as a valuable spontaneous large animal model for human Non-Hodgkin's Lymphoma (NHL). Herein we describe the establishment and characterisation of an in vivo model using the canine B-cell lymphoma cell line CLBL-1 analysing the stability of the induced tumours and the ability to resemble the original material. CLBL-1 was injected into Rag2(-/-)γ(c) (-/-) mice. The generated tumor material was analysed by immunophenotyping and histopathology and used to establish the cell line CLBL-1M. Both cell lines were karyotyped for detection of chromosomal aberrations. Additionally, CLBL-1 was stimulated with IL-2 and DSP30 as described for primary canine B-cell lymphomas and NHL to examine the stimulatory effect on cell proliferation. CLBL-1 in vivo application resulted in lymphoma-like disease and tumor formation. Immunophenotypic analysis of tumorous material showed expression of CD45(+), MHCII(+), CD11a(+) and CD79αcy(+). PARR analysis showed positivity for IgH indicating a monoclonal character. These cytogenetic, molecular, immunophenotypical and histological characterisations of the in vivo model reveal that the induced tumours and thereof generated cell line resemble closely the original material. After DSP30 and IL-2 stimulation, CLBL-1 showed to respond in the same way as primary material. The herein described CLBL-1 in vivo model provides a highly stable tool for B-cell lymphoma research in veterinary and human medicine allowing various further in vivo studies.
  PLoS ONE 01/2012; 7(6):e40078. · 4.09 Impact Factor
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  Available from: Sandeep S Patil

  Article: Virotherapy of Canine Tumors with Oncolytic Vaccinia Virus GLV-1h109 Expressing an Anti-VEGF Single-Chain Antibody.

  Sandeep S Patil, Ivaylo Gentschev, Marion Adelfinger, Ulrike Donat, Michael Hess, Stephanie Weibel, Ingo Nolte, Alexa Frentzen, Aladar A Szalay
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  ABSTRACT: Virotherapy using oncolytic vaccinia virus (VACV) strains is one promising new strategy for cancer therapy. We have previously reported that oncolytic vaccinia virus strains expressing an anti-VEGF (Vascular Endothelial Growth Factor) single-chain antibody (scAb) GLAF-1 exhibited significant therapeutic efficacy for treatment of human tumor xenografts. Here, we describe the use of oncolytic vaccinia virus GLV-1h109 encoding GLAF-1 for canine cancer therapy. In this study we analyzed the virus-mediated delivery and production of scAb GLAF-1 and the oncolytic and immunological effects of the GLV-1h109 vaccinia virus strain against canine soft tissue sarcoma and canine prostate carcinoma in xenograft models. Cell culture data demonstrated that the GLV-1h109 virus efficiently infect, replicate in and destroy both tested canine cancer cell lines. In addition, successful expression of GLAF-1 was demonstrated in virus-infected canine cancer cells and the antibody specifically recognized canine VEGF. In two different xenograft models, the systemic administration of the GLV-1h109 virus was found to be safe and led to anti-tumor and immunological effects resulting in the significant reduction of tumor growth in comparison to untreated control mice. Furthermore, tumor-specific virus infection led to a continued production of functional scAb GLAF-1, resulting in inhibition of angiogenesis. Overall, the GLV-1h109-mediated cancer therapy and production of immunotherapeutic anti-VEGF scAb may open the way for combination therapy concept i.e. vaccinia virus mediated oncolysis and intratumoral production of therapeutic drugs in canine cancer patients.
  PLoS ONE 01/2012; 7(10):e47472. · 4.09 Impact Factor
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  Available from: Cornelia Silaghi

  Article: Seroprevalence and bacteremia [corrected] of Anaplasma phagocytophilum in cats from Bavaria and Lower Saxony (Germany).

  Dietmar Hamel, Anna Bondarenko, Cornelia Silaghi, Ingo Nolte, Kurt Pfister
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  ABSTRACT: Anaplasma (A.) phagocytophilum is a tick-transmitted obligate intracellular bacterium and has been identified in a wide range of mammalian species, causing febrile disease in some. Few reports show that it can also cause granulocytic anaplasmosis in cats. As data on the occurrence of A. phagocytophilum in cats from Germany is limited, a total of 326 serum and 306 EDTA-blood samples from cats from Germany were screened by direct (Giemsa-stained blood/buffy coat smears, real-time PCR) and indirect (IFAT) methods. Of 274 Giemsa-stained blood smears which could be evaluated none was positive for morulae, but one blood sample (< or =0.1%; 1/306) was positive for A. < or = phagocytophilum-DNA in PCR. Antibodies (cutoff > or = 1:64) were detected in 53 out of 326 samples (16.2%). Altogether, the results show a high seroprevalence rate of anti-A. phagocytophilum antibodies in cats in Germany while the low detection rate of this bacterial agent by direct methods is similar to those of other studies on A. phagocytophilum infections in cats.
  Berliner und Münchener tierärztliche Wochenschrift 01/2012; 125(3-4):163-7. · 0.82 Impact Factor
• Article: Radiographic Evaluation of Early Periprosthetic Femoral Bone Contrast and Prosthetic Stem Alignment after Uncemented and Cemented Total Hip Replacement in Dogs.

  Ayman A Mostafa, Svenja Drüen, Ingo Nolte, Patrick Wefstaedt
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  ABSTRACT: OBJECTIVE: To radiographically evaluate periprosthetic femoral bone contrast and assess alignment of the prosthetic stem after uncemented and cemented total hip replacement (THR). STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n = 15). METHODS: Dogs were classified into uncemented (n = 8) and cemented (n = 7) THR groups. Radiographs were analyzed using image processing software to evaluate femoral bone contrast (gray scale value, GV) for each and combined modified Gruen zone(s) immediately and 4 months after THR. Modified Gruen zones were classified into 5 zones to analyze retrospectively the regional radiographic GV of the femur around uncemented and cemented prosthetic stem. Alignment of prosthetic stem was assessed immediately and 4 months postoperatively. Variables were compared by use of 2-tailed t-test, with P < .05 considered significant. RESULTS: Zone 1 showed significant decrease in the mean bone GV 4 months after uncemented THR. No differences in zones 1-5 after 4 months of cemented THR. Combined zones showed significant decrease in overall mean bone GV 4 months after uncemented THR. No changes were observed 4 months after cemented THR. Number of limbs with varus-aligned femoral stem markedly increased after 4 months of uncemented THR. CONCLUSIONS: Regional bone contrast and prosthetic stem alignment vary with the design of THR.
  Veterinary Surgery 12/2011; · 1.26 Impact Factor
• Article: TNF-α induced secretion of HMGB1 from non-immune canine mammary epithelial cells (MTH53A).

  Saskia Willenbrock, Olga Braun, Judith Baumgart, Sandra Lange, Christian Junghanss, Alexander Heisterkamp, Ingo Nolte, Jörn Bullerdiek, Hugo Murua Escobar
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  ABSTRACT: Mammary neoplasias are one of the most frequent and spontaneously occurring malignancies in dogs and humans. Due to the similar anatomy of the mammary gland in both species, the dog has become an important animal model for this cancer entity. In human breast carcinomas, the overexpression of a protein named high-mobility group box 1 (HMGB1) was reported. Cells of the immune system were described to release HMGB1 actively exerting cytokine function. Thereby it is involved in the immune system activation, tissue repair, and cell migration. Passive release of HMGB1 by necrotic cells at sites of tissue damage or in necrotic hypoxic regions of tumors induces cellular responses e.g. release of proinflammatory cytokines leading to elevated inflammatory response and neo-vascularization of necrotic tumor areas. Herein we investigated if a time-dependent stimulation with the separately applied proinflammatory cytokines TNF-α and IFN-γ can cause secretion of HMGB1 in a non-immune related HMGB1-non-secreting epithelial canine mammary cell line (MTH53A) derived from non-neoplastic tissue. The canine cell line was transfected with recombinant HMGB1 bicistronic expression vectors and stimulated after transfection with the respective cytokine independently for 6, 24 and 48 h. HMGB1 protein detection was performed by Western blot analysis and quantified a by enzyme-linked immunosorbent assay. Live cell laser scanning multiphoton microscopy of MTH53A cells expressing a HMGB1-GFP fusion protein was performed in order to examine, if secretion of HMGB1 under cytokine stimulating conditions is also visible by fluorescence imaging. The observed HMGB1 release kinetics showed a clearly time-dependent manner with a peak release 24h after TNF-α stimulation, while stimulation with IFN-γ had only small effects on the HMGB1 release. Multiphoton HMGB1 live cell microscopy showed diffuse cell membrane structure changes 29 h after cytokine-stimulation but no clear secretion of HMGB1-GFP after TNF-α stimulation was visible. Our results demonstrate that non-immune HMGB1-non-secreting cells of epithelial origin derived from mammary non-neoplastic tissue can be induced to release HMGB1 by single cytokine application. This indicates that tumor and surrounding tissue can be stimulated by tumor present inflammatory and necrotic cytokines to release HMGB1 acting as neo-vascularizing factor thus promoting tumor growth.
  Cytokine 12/2011; 57(2):210-20. · 3.02 Impact Factor
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  Available from: Cornelia Silaghi

  Article: Relationship of molecular and clinical findings on Anaplasma phagocytophilum involved in natural infections of dogs.

  Cornelia Silaghi, Barbara Kohn, Aleksandra Chirek, Claudia Thiel, Ingo Nolte, Gabriele Liebisch, Kurt Pfister
  Journal of clinical microbiology 12/2011; 49(12):4413-4. · 4.16 Impact Factor
• Article: BMP4 increases expression of HMGA2 in mesenchymal stem cells.

  Dominique Nadine Markowski, Burkhard Maria Helmke, Frauke Meyer, Inga von Ahsen, Rolf Nimzyk, Ingo Nolte, Jörn Bullerdiek
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  ABSTRACT: BMP4 has been linked to early steps of adipocyte lineage differentiation but only little is known about its corresponding downstream pathways. Herein, we have investigated whether or not the expression of high mobility group protein HMGA2, another protein linked to proliferation and differentiation within the process of adipogenesis, may be influenced by BMP4 signaling in adipose tissue derived stem cells. Compared to FGF1, a strong inducer of HMGA2 in immortalized pre-adipocytes, BMP4 was found moderately to induce the HMGA2 mRNA expression in serum starved adipose tissue derived stem cells and myometrial cells. In contrast, no such activity was noted in canine bone marrow derived mesenchymal stem cells. As to adipocyte lineage differentiation the functions of BMP4 and HMGA2 mechanistically overlap. Thus, we propose that in adipose tissue BMP4 acts in part by activating HMGA2 making this architectural transcription factor one of the major downstream players in that system.
  Cytokine 12/2011; 56(3):811-6. · 3.02 Impact Factor
• Article: Haemostatic abnormalities in cats with naturally occurring liver diseases.

  Brigitte Dircks, Ingo Nolte, Reinhard Mischke
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  ABSTRACT: Alterations in the haemostatic system were characterized in cats with different naturally occurring liver diseases. The study looked at 44 healthy cats and 45 cats with different liver diseases confirmed histologically or cytologically (neoplasia, n=9; inflammation, n=12; hepatic lipidosis, n=13; other degenerative liver diseases, n=11). The following parameters were evaluated: platelet count; prothrombin time; activated partial thromboplastin time; thrombin time; factor (F) II, FV, FVII, FX, and FXIII activities; fibrinogen concentration; activities of antithrombin, protein C, plasminogen, and α(2)-plasmin inhibitor, and D-dimer concentration. In cats with liver diseases, 44/45 (98%) had one or more abnormalities of the coagulation parameters measured. In cats with inflammatory liver diseases, increased D-dimer concentrations and decreased FXIII activity were the most consistent abnormalities and were found in 83% and 75% of cats, respectively. The most common abnormality in cats with neoplastic liver disease was FXIII deficiency (78%). The most consistent abnormalities in cats with hepatic lipidosis were increased FV activity and D-dimer concentration with 54% of cats having values above the reference range for both parameters. Cats with miscellaneous degenerative liver disease most frequently showed FXIII deficiency (64%). The results of this study show that alterations of single haemostatic components are a frequent finding in cats with liver disease. Activation of haemostasis with subsequent consumptive coagulopathy (rather than decreased synthesis) seems to be responsible for these alterations. Increased blood levels of different haemostatic components in cats with inflammatory lesions may be related to an acute phase reaction.
  The Veterinary Journal 11/2011; 193(1):103-8. · 2.24 Impact Factor