bluuebunny/crossref_metadata_2025_split huggingface dataset:
The Isolated Haemoperfused Distal Cow Limb as an Experimental Model for Studies on the Pathomechanisms of Bovine laminitis
Authors: Wüstenberg R. Y. , Mülling Ch. K. W. , Budras K. ‐D. 
Published: November 2005

Introduction:  The pathomechanisms of bovine laminitis are still poorly understood. Isolated perfused organ models are promising tools to study the biological mechanisms that lie behind diseases. They are preferable for ethical and financial reasons but also for experimental necessities.Aims:  To develop an isolated haemoperfused distal cow limb model, basing on an already existing porcine limb model used in experiments studying the effects of bioactive molecules.Methods:  Isolated limbs and blood were obtained from routinely slaughtered cows and subsequently perfused in the laboratory of a biotech company (Vitro‐Tec, Berlin) with expertise in organ perfusion. A standard perfusion apparatus enables perfusion for up to six hours under close to physiological conditions. Perfusion pressure and flow were calculated based on values available for horses and pigs. The oxygen saturation was kept at 100% and the pH between 7.35 and 7.45. Glucose was added as nutrient. The lactate concentration was measured as a parameter for oxygen supply and free haemoglobin as an indicator for haemolysis. Potassium content and pressure served as vitality indicators. Additionally, morphological criteria of tissue integrity were assessed using light and electron microscopy.Results:  A constant flow of 150–200 ml/min was optimal. The resulting pressure ranged from 30 to 117 mmHg. Light microscopy revealed, with exception of a few thromboses, no cellular damage after 4 h. The perfusion was successfully extended to 5 h. Then first pressure related alterations including vascular dilatations and cellular degeneration occurred in the dermal lamellae but not in the other regions. Potassium never exceeded physiological values.Conclusion:  The model is now available for experiments. Equipment and technical implementation have been standardized and described in a standard operating procedure. The model will be challenged by bioactive molecules such as lactate, neuropeptides and chemokines. Ischaemic conditions will be simulated and studied.This work was supported by the European Communities under the Lame Cow Project QLK5‐CT‐2002‐00969.The authors are solely responsible and the work does not necessarily represent the opinion of the European Communities.
A Potential Role for Platelet‐activating Factor (PAF) During Bovine Placentation
Authors: Bücher K. , Pfarrer C. , Leiser R. 
Published: November 2005

Platelet‐activating factor (PAF) is an important phospholipid messenger involved in diverse physiological and pathological conditions. Actions of PAF are mediated by a specific PAF receptor (PAF‐R) and the inhibition of biological activity by PAF acetylhydrolase (PAF‐AH). To elucidate the potential role of PAF in stimulating angiogenesis and trophoblast giant cell migration during gestation, we examined the expression and localization of PAF‐R mRNA and protein as well as PAF‐AH mRNA in the bovine placenta. RT‐PCR, in situ hybridization and immunohistochemistry were applied to placentomes from days 20, 80, 150, 200, 240, 270 of gestation, and immediately pre‐partum (around day 280). PAF‐R mRNA and protein were colocalized in immature trophoblast giant cells (TGC), uterine epithelial cells, and endothelial cells of fetal and maternal vessels. There was no apparent difference in placental PAF‐R mRNA or protein levels until day 270. In contrast, in placentomes taken immediately pre‐partum the localization of the immunohistochemical and hybridization signals switched from immature TGC and maternal epithelium to cells of the maternal stroma, particularly endothelial cells of maternal vessels. mRNAs for PAF‐AH isoforms Ib (β‐ and γ‐subunit), II, and plasma‐isoform were detected by RT‐PCR in placentomes throughout gestation. In tissue sections mRNA of the PAF specific PAF‐AH‐Ib β‐subunit was predominantly found in TGC, while the intensity of the signal was weaker in maternal epithelium and endothelium of fetal and maternal vessels. Corresponding to the PAF‐R expression, mRNA of PAF‐AH Ib was detected mainly in maternal endothelial cells prior to parturition. The constitutive expression and localization of PAF‐R and PAF‐AH in epithelial and endothelial cells during gestation, together with the parturition‐related switch to the maternal stroma suggests a functional role of PAF for the interaction of TGC with the maternal epithelium, and thus may be associated with TGC migration, placental angiogenesis and pre‐partum tissue programming.Acknowledgement:  This study was supported by the German Research Foundation.
Expression and Role of Connexin Proteins in Bovine Claw Epidermis
Authors: Frohberg‐Wang D. , Mülling Ch. K. W. , Budras K. D. 
Published: November 2005

Gap junction mediated inter‐cellular communication plays a crucial role in maintaining tissue homeostasis in skin. Gap junctions consist of hemi‐channels (connexons), which in turn are established by transmembrane proteins (connexins, Cx). The pattern of Cx‐ protein expression is characteristic for tissues and depends on the differentiational and functional state.Aim:  As part of our investigations of cell biological causes for lameness in dairy cows we wanted to assess the expression of Cx‐proteins in relation to differentiation in bovine claw epidermis.Methods:  We examined hoof tissue from five defined regions of the claws of 10 cows from the abattoir by immunohistochemistry, Western blot and by transmission electron microscopy. The following monoclonal antibodies were used: anti‐ Cx26, Cx31, Cx37 (ADI, USA) and anti‐Cx43 (Chemicon, USA). Immunohistochemistry was carried out on frozen and paraffin sections.Results:  Cx43 was expressed in the bovine claw epidermis in a pattern described for skin. Cx43 expression was strongest in the stratum basale in all regions. Cells of the stratum spinosum expressed lower levels, and in terminal differentiating cells Cx43 was disappearing. Expression of Cx26, 31 and 37 showed a high variation between individuals and regions. Protein expression was verified by Western blotting. The formation of gap junctions in the respective layers was confirmed by electron microscopy.Conclusion:  Our results provide evidence that Cx 43 is expressed in bovine claw epidermis in a distribution similar to skin. Cx 43 is progressively downregulated with the progress of differentiation. This suggests a role of Cx43 in cellular coordination particularly in the early stages of differentiation. Results for Cx 26, 31 and 37 will be confirmed in further studies also on the mRNA level to get deeper insight in the regulation of Cx expression in claw epidermis. We hypothesize that Cx 43 plays a similar role in controlling differentiation as in skin and may be involved in disruption of claw horn formation in claw disorders.Acknowledgement:  This work was supported by the European Communities under the Lamecow project QLK5‐CT‐2002‐00969. The authors are solely responsible and the work does not necessarily represent the opinion of the European Communities.
Morphological and Ultrastructural Study of Vaccine‐induced Granuloma in Atlantic Salmon (Salmo salar)
Authors: Haugarvoll E. , Bjerkås I. , Poppe T. T. , Koppang E. O. 
Published: November 2005

Aim:  The aims of this study were immunhistochemical (IHC) and ultrastruktural characterization of cells involved in vaccine granuloma formation in Atlantic salmon.Methods:  Fish suffering from severe reaction 6 to 9 months after vaccination were selected. We investigated granuloma from peritoneum, at the injection site. Morphological staining techniques, IHC to visualise MHC+ cells and ultrastructural analysis were performed.Results:  Microbial investigations revealed no infective agents. Histological and ultrastructural studies revealed Russel bodies, macrophages, multinucleated giant cells, melanomacrophages (MMP), mast cells (MC) and Splendore–Hoepply material in the lesions.Conclusion:  Signet ring cells identified after vaccination have been hitherto described as svollen mast cells. This study shows that these cells fit the Russel body characteristic, consistent with humoral stimulation following vaccination and foreign material precipitate, Splendore‐Hoepply material. MHC+ cells were located circumferal to granuloma centers. MMP were present containing melanosomes in different developmental stages.
How to Open the Mouth: the Kinetic Skull, and the Jaw and Branchial Apparatus in Sharks
Authors: Homberger D. G. , Westphal K. A. , Gudo M. , Bragulla H. H. 
Published: November 2005

Aims:  Mammals cannot swallow if their necks and, hence, their mm. sternohyoidei et sternothyroidei are stretched. In order to understand the functional morphology of mastication and swallowing in mammals, it is useful to understand its evolutionary origin.Material and Methods:  Five formalin‐preserved specimens of the Spiny Dogfish (Squalus ancanthias) were used for dissection or to prepare sagittal or transverse serial sections.Results and Conclusions:  The feeding apparatus of the shark Squalus acanthias as a representative of the cartilaginous fishes serves as an excellent model of that of an ancestral vertebrate with a kinetic skull, in which the chondrocranium is movable relative to the jaw apparatus. To open the mouth, the cranium is raised by the epibranchial musculature, the palatoquadrate cartilage (i.e., upper jaw) is entrained, and the mandibular cartilage is held in place and prevented from following the upper jaw by the coracomandibular muscle. At the same time, the branchial basket and oropharyngeal cavity are expanded and held in place by the coracohyoid and coracoarcual muscles, which prevent the hyoid and branchial arches from being entrained cranially by the raised chondrocranium. The mandible is not retracted during the opening of the mouth in sharks because the branchial region would have to be compressed instead of expanded. In the course of the evolutionary modifications leading to mammals, the heart and respiratory apparatus became decoupled from the feeding apparatus through the formation of a neck. The skull became akinetic through the integration of the upper jaw into the base of the cranium. The mouth is not opened any longer by lifting the cranium, but by depressing the mandible through a special muscle, the m. depressor mandibulae. The coracomandibular muscle lost its function as a stabilizer of the mandible during mouth opening and became modified into the lingual and geniohyoid muscles.
Fish immunology. Fish Erythrocytes Release Cytokine‐like Factors that Modulate Autologous Macrophage Function
Authors: Jirillo E. , Passantino L. , Ribaud M. R. , Passantino G. F. 
Published: November 2005

According to our previous findings (Passantino et al. 2002a,b; 2004) we discovered that fish erythrocytes (FE) act as immune cells in that they bind and engulf Candida albicans (CA). Then, these activated FE interact with macrophages (MØ) so that a so‐called rosette results from such an interplay. A rosette consists of one or more MØ containing ingested CA, centrally located and surrounded by CA activated FE. Quite interestingly, we found that supernatants of CA‐activated FE contain cytokine‐like factors able to modulate MØ function. In particular, active supernatants are able to inhibit MØ migration under agarose and this activity was identified as macrophage inhibiting factor (MIF) – like function, by analogy to human MIF. On the other hand, same supernatants could enhance MØ phagocytosis of CA, thus contributing to fungus killing. Taken together, these results suggest that FE act as immune cells by secreting cytokine‐like factors, which contribute to the rosette formation through modulation of MØ.References  Passantino, L., M. Altamura, A. Cianciotta, R. Patruno, A. Tafaro, E. Jirillo, and Passantino G.F., 2002a: Fish immunology. I. Binding and engulfment of Candida albicans by erythrocytes of rainbow trout (Salmo gairdneri Richardson). Immunopharmacol. Immunotoxicol. 24: 665–678Passantino, L., A. Tafaro, M. Altamura, R. Arena, G.F. Passantino, E. Jirillo, 2002b: Fish immunology. II. Morphological and cytochemical characterization and phagocytic activities of head kidney macrophages from rainbow trout (Salmo gairdneri Richardson). Immunopharmacol. Immunotoxicol.24: 679–691Passantino, L., M. Altamura, A. Cianciotta, F. Jirillo, M.R. Ribaud, E. Jirillo, and G.F. Passantino, 2004: Maturation of fish erythrocytes coincides with changes in their morphology, enhanced ability to interact with Candida albicans and release of cytokine‐like factors active upon autologous macrophages. Immunopharmacol. Immunotoxicol. 26 (in press).
A Novel In Vitro Model of Canine Malignant Hemangioendothelioma
Authors: Kühn D. , Kohn B. , Brunnberg L. , Plendl J. 
Published: November 2005

Introduction and Aim:  Canine malignant haemangioendothelioma is an aggressive neoplasia that affects mostly older dogs of large breeds with a strong predilection for the spleen, liver, heart and skin. The tumour originates in the vascular endothelium and consists of transformed cells forming large and leaky vessel‐like structures. Prognosis is poor because surgery and chemotherapy have limited success in prolonging survival times and increasing quality of patients. A new strategy to treat this malignancy could be anti‐angiogenic therapy based on the inhibition of proliferation, migration and three‐dimensional organization of transformed cells. In order to reduce animal experiments, in vitro‐models are required to test the safety and efficacy of anti‐angiogenic drugs. So far only few models of angiogenesis are available using mostly human, rodent and bovine cells. Therefore, the aim of our study was to establish an in vitro model of canine haemangioendothelioma.Materials and Methods:  Tumours were collected from dogs during surgery or immediately after euthanasia. Isolation of cells was done from different areas of the tumours and by enzymatic digestion of the tissue. Cells were incubated in culture media with and without endothelial growth factors. Cells were characterized by lectin histochemistry using Dolichos biflorus agglutinin, Ulex europaeus agglutinin and Bandeiraea simplicifolia agglutinin I. Moreover, RT‐PCR (polymerase chain reaction) was employed to investigate the expression of vascular endothelial growth factor (VEGF) and its endothelium‐specific receptors VEGF‐R1 and ‐R2.Results and Conclusions:  Different populations of cells were isolated and cultured successfully from canine malignant haemangioendothelioma. Cells show characteristics of microvascular endothelial cells of an angiogenic phenotype, i.e. the formation of spheroids and tube‐like structures as well as strong labelling for Bandeiraea simplicifolia agglutinin I. Thus, morphological and glycohistochemical results confirm the vascular character of the cells isolated. RT‐PCR showed expression of VEGF. However, endothelium‐specific VEGF receptors were not expressed. Loss of typical receptors is common in cancer and may correlate with increased tumour dedifferentiation.
Regulation of Cellular Proliferation and Differentiation in Claw Epidermis by Epidermal–Dermal Interactions
Authors: Mülling Ch. K. W. , Nebel U. , Budras K. ‐D. 
Published: November 2005

Introduction:  Epidermal–dermal interactions play an important role in regulation of proliferation and differentiation of keratinocytes and also in disease and regeneration of skin. A double paracrine regulation of keratinocyte growth and differentiation has been postulated and described in vitro recently. Interleukin‐1 (Il‐1) and keratinocyte growth factor (KGF/FGF‐7) and their receptors are major mediators in this epidermal‐dermal signalling.Aim:  We wanted to generate evidence for a reciprocal regulation of epidermal proliferation and differentiation in the bovine claw involving Il‐1 and KGF.Methods:  Tissue samples from claws of slaughtered cows, co‐cultures and three‐dimensional cultures of bovine hoof keratinocytes and fibroblasts were used. Immunohistochemistry for Il‐1 and Il‐1 receptor, KGF and KGF receptor were carried out on frozen and paraffin sections from tissue and cell culture. RT‐PCR was used to study regulation at the mRNA level in dermis and epidermis (after micro separation) and in fibroblasts and keratinocytes from the cultures respectively.Results:  In claw epidermis Il‐1 was accumulated and KGF receptor was detectable. Interleukin‐1 receptor and KGF were present in dermal cells. Il‐1 stimulated fibroblasts responded with KGF expression in vitro. Neutralization of Il‐1 reduced or stopped this effect. Keratinocytes co‐cultured with fibroblasts showed a significant better growth. Presence of keratinocytes increased KGF expression in the co‐cultured fibroblasts.Conclusion:  Our results suggest that Il‐1 is leading to stimulation of KGF expression. KGF seems to be an important stimulator of keratinocyte proliferation in our co‐culture system. So far our results provide initial evidence for a reciprocal regulatory mechanism present in the bovine claw involving Il‐ 1 produced in the epidermis and KGF originating from fibroblasts. Future in vitro studies including mRNA analysis will provide deeper insight into the epidermal‐dermal cross‐talk and the involved transcriptional/translational regulation in the bovine claw.This work was supported by the European Communities under the Lame cow project QLK5‐CT‐2002‐00969. The authors are solely responsible and the work does not necessarily represent the opinion of the European Communities.
3D‐Reconstructions: New Application Fields in Modern Veterinary Anatomy
Authors: Nöller C. , Henninger W. , Grönemeyer D. , Budras K. ‐D. 
Published: November 2005

Introduction:  Today, the claims on the anatomical education have changed with the considerable increase of modern medical imaging. For the analysis of computed tomography (CT) images, magnetic resonance (MR) images or ultrasound, the knowledge of cross sectional anatomy is an important prerequisite for proper diagnosis. Admittedly, it is difficult for students and also for practitioners to imagine three‐dimensional objects in space. The possibilities and convenience of 3D‐reconstructions were demonstrated by generating virtual casts of frontal sinuses.Methods:  Using a HiSpeed (DX/i Plus, General Electric) scanner transversal CT images of Persian and European Shorthair cats were acquired. The slice thickness and distance were set to 1 mm. With the software system VG Studio MAX 1.1, the obtained volumetric data was used to create volume graphics displaying object surfaces and interiors based on voxel data. Because of density value acquisition, it was possible to identify and label the objects of interest. The extension of the paranasal sinuses was analyzed and demonstrated in relation to the skull. For verification, conventional corrosion casting employing Technovit or Tensolcement, respectively, were carried out on the same specimens.Results:  The frontal sinuses of both feline breeds showed great variations in size, shape and symmetry. With models from the skull based on 3D‐datasets including the virtual casts of the paranasal sinuses, it was easy to imagine spatial relations and localizations inside the head. The advantages of the virtually derived casting method are: creating the virtual casts in vivo; the method does not imply any health risks and is repeatedly applicable.Conclusion:  3D‐reconstructions based on CT‐datasets are excellently suited for generating spatial sense. Additionally, a single scan series can be applied for different anatomical problems. The resulting digital 3D‐objects are readily available and can be employed for educational purposes for better comprehension of the difficult spatial relations of the head.
The Origin of Melanomacrophages Centers in Salmo Gairdneri Richardson
Authors: Passantino L. , Ribaud M. R. , Cianciotta A. , Altamura M. , Massaro M. A. , Passantino G. , Jirillo E. 
Published: November 2005

By definition, melanomacrophage centres (MMC) consist of aggregates of macrophage‐like pigmented cells located in the kidney and spleen, and to a lesser extent in the liver. In 30 Salmo gairdneri Richardson trout, MMC have been identified in various organs. In the kidney, liver and spleen, respectively, macrophages (MØ) are aggregated in clusters, which are next to vessels. MMC have been cytoenzymatically identified with periodic acid Schiff (PAS), α‐naphthyl acetate esterase (ANAE), peroxidase (PEROX) reactions. Finally, in order to show pigments in the context of MMC, in particular, we used iron stain to detect iron pigments and Prussian blue reaction to detect melanin and lipofuscin. The presence of iron pigments indicates that MMC function as lymphoreticular centres where fish erythrocytes (FE) are destroyed together with other foreign material. In previous studies, we found that FE are able to bind and engulf Candida albicans (CA) without killing this fungus. FE containing engulfed CA form with MØ functional rosettes and, in reason of that MØ are solicited to phagocytose and kill CA. It is likely that rosettes and/or single FE containing engulfed foreign material migrate to distant organs, such as the kidney, the spleen and the liver. Therefore, the MMC may, in part, derive from these peripheral rosettes and pigments result from FE and foreign material digestion and destruction operated by MØ. Reference  Passantino, L., Cianciotta, A., Jirillo, F., Carrassi, M., Jirillo, E. and Passantino, G.F. 2005: Lymphoreticular system in fish: erythrocyte‐mediated immunomodulation of macrophages contributes to the formation of melanomacrophage centers. Immunopharmacol Immunotoxicol. 27, 147–161.
Rhizoctonia solani AG 2‐1 as a causative agent of cotyledon rot on European beech (Fagus sylvatica)
Authors: Hietala A. M. , Mehli L. , Nagy N. E. , Kvaalen H. , La Porta N. 
Published: November 2005

Summary Rhizoctonia solani was frequently isolated in the Italian Alps from nursery‐grown European beech (Fagus sylvatica) seedlings displaying symptoms of cotyledon rot. Koch's postulates were verified and mode of infection of the associated isolates was investigated with light and scanning electron microscopy. Population structure of the pathogen was investigated by scoring the anastomosis reaction type in pairings between different isolates from the same seedbed. One pathogen genotype showed a large distribution area within the seedbed, this implying that the inoculum had been building up in the seedbed over a longer time period. Hyphal anastomosis tests and sequence analysis of the internal transcribed spacer (ITS) region of ribosomal DNA indicated that the pathogen belongs to AG 2‐1 of R. solani. ITS sequence analysis indicates that the isolates from beech are closely related to R. solani isolates causing a disease on tulip. The striking similarities between the two diseases are discussed.
Spontaneous uterine rupture at 35 weeks’ gestation, 3 years after laparoscopic myomectomy, without signs of fetal distress
Authors: Banas Tomasz , Klimek Marek , Fugiel Andrzej , Skotniczny Krzysztof 
Published: November 2005

AbstractLaparoscopic myomectomy (LM) is a recently developed surgical technique, and every obstetrician should be aware of its possible complications, which can occur not only during labor but also during pregnancy. We report a case of a primigravid woman who was hospitalized at 35 weeks’ gestation because of irregular abdominal pain. She conceived spontaneously 3 years after LM. After a 20‐h stay on the obstetrician ward due to increased abdominal tenderness and vaginal bleeding, the patient was qualified for an emergency cesarean section without any symptoms of fetal distress in cardiotocography. During cesarean section a newborn with 9 Apgar points was delivered and a rupture of the uterine wall was seen. Dehiscence of the pregnant uterus following LM is an incidental case, and can therefore be misdiagnosed. Close attention should be paid to every pregnancy in previously operated uteri as the dehiscence of the pregnant uterus can occur without symptoms of fetal distress.