bluuebunny/crossref_metadata_2025 huggingface dataset:
Anatomical Location and Histology of the Tonsilla Paraepiglottica in Sheep
Authors: Cocquyt G. , Simoens P. , Van den Broeck W.
Published: November 2005

The location, size and histological structure of the ovine paraepiglottic tonsils were determined in 20 sheep. Macroscopic examination of the pharyngeal tonsils was performed in 10 sheep of 9 months of age, of which the heads were fixed in acetic acid 2% for 24 h, and in non‐fixed heads of five sheep aged between 6 and 9 months. The paraepiglottic tonsil is located lateral to the epiglottis in the ventromedial segment of the pharyngeal sidewall, i. e. at the floor of the piriform recess. The tonsil appears as a cluster of small nodular elevations, which are white after fixation in acetic acid and are called hereafter tonsillar nodules. The number of these nodules per tonsil ranged from 1 through 8. The length of the tonsil varied from 5 till 14 mm, and the width ranged from 2 till 5 mm. The tonsils of 5 sheep aged between 6 and 9 months and of five sheep aged between 9 and 12 months were excised and fixed in 3.5% formaldehyde. The blocks were processed routinely for histology and embedded in paraffin wax. Serial sections (8 μm thick) were stained with eosin and haematoxylin. Histology revealed that each tonsillar nodule consisted of several lymphoid follicles (noduli lymphatici) surrounded by parafollicular tissue. Most lymphoid follicles were secondary follicles consisting of a germinal centre, a dark zone and a mantle zone. Primary follicles devoid of germinal centres were most obvious at the periphery of the tonsillar nodules. The stratified squamous pharyngeal epithelium covering the lymphoid tissue was frequently infiltrated by numerous lymphocytes, forming a follicle‐associated epithelium. Crypts were absent, but in some cases the pharyngeal epithelium was slightly infolded into the lymphoid tissue. The tonsillar nodules were almost completely encapsulated by dense connective tissue from which septa extended into the larger nodules.

Acknowledgement:  This study was supported by BOF Grant of Ghent University Project #011B4101).
Teaching Developmental Anatomy of the Genital System to VM1 Students at the University of Missouri‐Columbia, USA
Authors: Constantinescu I. A.
Published: November 2005

Developmental anatomy is integrated with gross anatomy. It consists of 12 lectures and is divided into body systems. The development of each system is correlated to its adult structures and species differences. The urinary and genital systems are developmentally and anatomically associated and originate together from the intermediate mesoderm. The urethra forms from the distal part of the urogenital sinus. Starting with the gonadogenesis, and continuing with the duct systems and external genitalia, the lectures are focused on the male and then on the female structures. At the end, the anomalies of the genital organs are exposed in detail, with suggestions for students, specialists and breeders to prevent and treat teratological conditions. The gonadogenesis is first presented in the indifferent stage. The differential stage is explained for the testis and then for the ovary. The genital duct system is exposed in the indifferent stage. The differential stage for the male includes the formation of the efferent ductules, the epididymal duct and the ductus deferens, while in the female, the focus is on the formation of the uterine tubes, uterus (including the horns, the body and the cervix), and the vagina. The indifferent stage of external genitalia is a complex stage, which originates from the mesodermal swellings adjacent to the cloacal membrane. In the male, the differential stage includes the formation of penis and penile urethra. In the female the genital tubercle forms the clitoris, the urethral folds form the labia, while the genital swellings degenerate. The hermaphroditism, gonadal dysplasia, the cryptorchidism, the persistent vestigial structures, and the penischisis (hypospadias and epispadias) as anomalies of the genital system conclude the lecture.
Comparison of the Comformation of 20 International Level and the 20 Low Level Jumping Horses Using a 3‐D Video Morphometric Measurement Method
Authors: Crevier‐Denoix N. , Pourcelot P. , Erlinger D. , Concordet D. , Lagache C. , Ricard A. , Tavernier L. , Denoix J. M.
Published: November 2005

A new method of morphometric measurement has been developed (method presented as a poster at the 2002 EAVA congress). Its main originalities are (1) to use video (each horse is filmed while walking instead of being photographed at standing), (2) to be in 3‐D (4 digital cameras are used; they film the horse from its lateral side, and also from the front and rear), (3) not to require skin markers (which makes the recording procedure quicker). This method is presently applied in the context of the breeding program of the Selle Français (SF) horse, through two large sample studies. Before starting the analysis of the corresponding data, a preliminary study was performed in order to settle the analysis methodology and to select the most pertinent morphometric parameters to characterize the “model” (conformation) of sport horses. Twenty international level SF horses (jumping performance index (ISO) >= 150, group A), and 20 low‐level (class D) jumping horses (group B) were compared using the 3‐D method. Out of the 400 morphometric parameters calculated for each horse, about 132 revealed significant differences (Student test, P < 0,05) between the 2 groups; P was < 0.001 for 26 of these parameters. The height of the main anatomical landmarks of the pelvic limb, acting as essential lever arms for propulsion muscles (ex: point of the hock (tuber calcanei), ischial tuber), and the height – absolute and relative (to the withers height) – of the croup, the length of the croup and thigh, the length of the fore cannon, the shortness of the forearm, and the relative shortness of the trunk, are all conformational traits that characterize group A compared with group B.
Detection of Steroid Hormone Receptors in the Canine Circumanal Gland
Authors: Egerbacher M. , Spillner A. , Weissenböck H.
Published: November 2005

The morphology and function of the circumanal glands in dogs is under current discussion. They are clinically important because they rank fourth in frequency as the site of canine skin tumours. The tissue is known to be governed by steroid hormones, as the gland exhibit sexual dimorphism, castration reduces the size of the gland and lead to tumour regression. The aim of our study was to demonstrate the hormone receptors in healthy circumanal glands as well as adenomas and adenocarcinomas in intact and castrated female and male dogs of different age groups. Immunohistochemical staining was performed using antibodies against androgen receptor (AR, Affinity Bioreagents, rabbit pc, 1:150) and oestrogen receptor (ER, Zymed Lab., rabbit pc, 1:200) after antigen retrieval with microwave cooking. Positive and negative nuclei were counted using Eclipse software and subsequently statistically evaluated. ER and AR‐positive nuclei could be detected in healthy as well as in diseased tissue sections of the circumanal gland. The number of positive nuclei was higher in the flattened peripheral cells compared to the large polygonal cells in the centre of the lobule. There was no significant difference between male and female tissue and between healthy glands and tumours although in tumours there was a tendency to a higher number of positive cells. In tumours, the distribution of hormone receptor positive cells was more irregular and varied between single lobules. Castrated animals of both sexes showed a slightly higher number of positive nuclei than intact animals, but the difference was not significant. Our results demonstrate for the first time the presence of steroid hormone receptors in the canine circumanal gland. We suggest that an evaluation of steroid hormone receptors is justified before general hormonal or anti‐hormonal treatment of patients.
Specific Localization of the Gap Junctional Protein Connexin 32 in Equine Gastric Mucosa
Authors: Fink C. , Hembes T. , Brehm R. , Weigel R. , Heeb C. , Tatar M. , Kressin M.
Published: November 2005

Gap junctional inter‐cellular communication (GJIC) is known to be important in the maintenance of tissue homeostasis. Evidence is accumulating that GJIC plays an important role in the gastric mucosal defence system and that the loss of GJIC is associated with gastric ulcer formation. The prevalence of gastric ulceration in foals and horses is high. However, there are no studies about the presence of gap junctions in gastric mucosa of horses. The purpose of this study was to investigate the distribution pattern of the gap junction protein 32 in normal gastric mucosa of horses. Immunocytochemical observations were made with light microscopy on cryosections of fresh frozen gastric mucosa from the fundic region, pyloric region, margo plicatus and pars nonglandularis. Immunohistochemical staining was performed by standard immunoperoxidase techniques using an anti‐connexin 32 polyclonal antibody. In Situ Hybridization, Western Blot Analysis and RT‐PCR were performed to confirm the presence of connexin 32. In normal horse gastric mucosa, connexin 32 was abundant in the surface epithelium and in a decreasing staining gradient extending down to the neck of the glands. Gastric surface mucous cells are formed in the neck of the glands and migrate along the foveola toward the mucosal surface. So, immature forms of surface mucous cells are found in the generative zone and in the deep part of the foveola, whereas well‐matured ones cover the upper part of the foveola and the luminal surface of the mucosa. Therefore, these findings are in line with other studies, which described larger and more numerous gap junctions in mature surface mucous cells than in immature cells. These results show that gap junctions develop during the maturation of surface mucous cells and suggest that GJIC between gastric surface mucous cells plays an important role in the regulation of cell differentiation and in tissue homeostasis. Further studies are required to investigate the distribution pattern of connexin 32 in the gastric mucosa of horses with gastric ulcer.
Anatomical Basis of Physiotherapeutic Treatment of Lumbar Spinal Nerve Disorders
Authors: Grünenfelder F. I. , Steffen F. , Mouwen M. A. , Boos A.
Published: November 2005

In contrast to human medicine, the significance of appropriate post‐operative care in companion animals is often neglected or even unknown. Physiotherapeutic exercises, also called physical therapies, are the golden standard of rehabilitation in human orthopaedics. In dogs and cats, such supportive therapies following surgery are rarely performed. Presently, physical therapeutic exercises, which are well established in man, are transferred to companion animals. However, there is no generally accepted scheme of procedures adequate to cats and dogs. Furthermore there is no study dealing with the exact effects of the various exercises performed with these species. Due to this paucity of information, the effect of several physical exercises concerning the lumbar spinal nerve roots transferred from man to the dog were assessed macroscopically. In ten euthanized healthy beagles up to eight years of age, the lumbar spine was surgically separated from adhering dorsal muscles. The lumbar nerve roots were visualized by removing the connective tissues surrounding the foramen intervertebrale. Nerve roots, i.e. spinal ganglia, were marked immediately outside the vertebral column applying a dot of yellow paint while the cadaver was in lateral recumbency. In man, for maximal sciatic nerve root movement the straight leg raising (SLR) (Maitland, 1986) exercise is applied on the patient lying on the back. For nervus femoralis movement the passive knee bending exercise is performed. In the dog, however, performing only the transferred SLR has a poor effect on L7. It can greatly be enhanced by a supplemental inward rotation of the hip joint by fixing the stifle joint and moving the lower leg medially. Concerning L4‐L6 nerve roots, the hip joint is maximally extended and the effect can be improved by an outward rotation of the leg. The results of the present study indicate that the simple transfer of physical exercises from man to companion animals does not result in identical results in the latter species. Modifications considering different anatomical conditions should be kept in mind if clinical results are poor.

Reference  Maitland, G.D., 1986: Vertebral Manipulation, 5th edition, Butterworth–Heinemann, London
New Concepts for the Functional Maturation of the Bovine Claw: Endothelial Biology, Angiogenesis and Ageing
Authors: Hirschberg R. M. , Müller K. , Plendl J.
Published: November 2005

Introduction:  Endothelial dysfunction and impaired angiogenesis are connected to ageing. However, the mechanisms involved in age‐related alterations are not fully elucidated, whereby reduced VEGF‐expression and decreased mobilization of circulating endothelial progenitor cells are suggested as possible causes. When cultured in vitro, endothelial cells often enter a growth arrest phase called ‘cellular senescence’ that is consistent with many changes in age‐related vascular disease. Based on the decreasing lifespan observed in high yielding dairy cows, a hypothesis has been formulated that these cows, due to their enormous metabolic performance, show an accelerated process of ageing, resulting in typical production‐related diseases, particularly lameness. As a basis for further research into the role of endothelial cell biology in high producing cattle, the aim of this study was to set up a model of endothelial ageing. As a first step, the role of angiogenesis in the functional adaptation of the microvasculature of the bovine pododerma was examined.

Material and methods:  Scanning electron microscopy of vascular corrosion casts of healthy juvenile and adult claws as well as pathologically altered claws, and routine light microscopy of pododermal serial sections was compared to an in vitro model of angiogenesis based on cultured bovine microvascular endothelial cells. For detection of angiogenic molecular control mechanisms effective in both, the pododermal system and our cell culture model, lectin‐ and immunohistochemistry as well as RT‐PCR were carried out.

Results and discussion:  In the adolescent and adult claw, sprouting and intussusceptive angioadaptation was demonstrated. Evidence of angiogenic processes was also detected in the pathologically altered claws. Immunolocalization of angiogenic factors within the dermo‐epidermal system of the claw was established. The detected angioadaptation may be visible expression of the increased metabolic demands of the claw caused by the growing body‐weight‐load. Increase of the pododermal vascular density via angiogenesis was also the connectional reparative principle in pathologically altered claws.
Long‐term Cultivation of Bovine Hoof Cells in a Novel Perfusion Chamber System
Authors: Hoffmann D. , Nebel U. , Mülling Ch. , Budras K. D.
Published: November 2005

The aim of this study was to establish a co‐culture of bovine hoof keratinocytes and fibroblasts in different types of perfusion chambers under defined conditions. The perfusion chamber PCS3c (Oligene, Berlin) was used to culture dermal fibroblasts and epidermal keratinocytes separated by a Millicell® Insert (Millipore, Schwalbach). In addition novel perfusion chambers developed by Dirk Hoffmann were used in combination with S & S membrane filters (Schleicher & Schuell, Dassel). First, fibroblasts were seeded onto one side of the insert/membrane and allowed to grow for 4 days. Subsequently keratinocytes were seeded onto the other side and allowed to adhere for one day. Then the chambers were connected to the tubing system with an attached 8‐channel pump and a gas permeable media bag. We worked with different media and flow rates ranging from 0.035 ml/min to 0.35 ml/min. The system was run for up to 20 days. After a few days in culture the cells had grown to confluence. Then keratinocytes began to differentiate and built up stratified colonies. Within these colonies the cells showed the characteristic morphology of a stratified squamous epithelium. The use of perfusion chambers allows three‐dimensional cultures to grow and survive for weeks, because of an unlimited medium supply and gas exchange. Additional, perfusion chamber systems enable the co‐cultivation of fibroblasts and keratinocytes separated by membranes, which permit the exchange of molecules like growth factors. Furthermore, it is possible to perfuse the two different parts of the perfusion chamber with various and even different types of media. This provides the opportunity to add a growth factor to the medium for only one cell type and to study the effect of this factor to the other cell type. 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.
Angiogenesis and Vascular Regression in the Corpus Luteum Periodicum
Authors: Hünigen H. , Bisplinghoff P. , Plendl J.
Published: November 2005

Introduction and Aim:  The corpus luteum is one of the most intensely vascularized tissues. Luteal angiogenesis is strictly controlled and blood vessels regress completely within a short period of time. The aim of this study was to investigate vascular dynamics in relation to cellular and molecular mechanisms of luteal angiogenesis and anti‐angiogenesis.

Material and Methods:  Endothelial cells of blood vessels in paraffin sections of bovine corpora lutea from different stages were examined by labelling with the lectin Bandeiraea simplicifolia agglutinin I. Angiogenesis was studied by morphometry of the capillaries, and immunolocalization of the angiogenic factor VEGF and VEGF‐receptor 2. Presence of apoptotic luteal and endothelial cells was investigated using the TUNEL test and transmission electron microscopy.

Results:  During development of the corpus luteum (day 3–8 of the oestrous cycle) a dense capillary network (8–12% area ratio) is established and maintained until day 17. Early regression (day 18–24) is characterized by a remarkable decrease of capillaries (1% area ratio). In the regressing corpus luteum the number of apoptotic luteal cells is closely correlated (r = 0.9) to the number of apoptotic endothelial cells. VEGF is immunolocalized in luteal cells (day 3–17), smooth muscle cells and endothelial cells of arterioles of the regressing corpus luteum. During late luteal regression, a moderate increase of capillaries (2.5% area ratio) is obvious.

Conclusions:  The dynamic changes of the capillarity during development and regression of the cyclic corpus luteum correlate with VEGF and VEGF‐R2 activities. In contrary to expectations the late stage of luteal regression is accompanied by angiogenesis. One reason for this phenomenon may be an increase in metabolic activity resulting in re‐organization of blood vessels already regressed.
Angiogenesis and Interstitial Pressure in a Rat Tumour Model
Authors: Hünigen H. , Drees R. , Schnorr J. , Plendl J.
Published: November 2005

Introduction and Aim:  Angiogenesis, the formation of new blood vessels, is a crucial process in physiological and pathological growth. Pathological angiogenesis is responsible for growth and metastasis of solid tumours, and, when blocked, improves prognosis. As a result of the angiogenic cascade in solid tumours an irregular, leaky capillary network develops. The aim of the present study was to define malignant tumours’ vascular characteristics and reveal functional anatomy by quantification of the microvasculature and interstitial pressure (IP) in relation to tumour fluid dynamics as visualized by contrast enhanced magnetic resonance imaging (MRI).

Material and Methods:  Dynamic MRI and measurement of the IP was performed in 21 rats implanted with colon carcinomas subcutaneously. Angiogenesis was studied by morphometry of the capillaries, and immunolocalization of the angiogenic factor VEGF and VEGF‐Receptor 2.

Results and Conclusions:  Histology, immunohistochemistry and MRI confirmed concentric arrangement of 4 tumour zones. The tumour margin included loose connective tissue with abundant mononuclear cells. Many large microvessels were seen in this most intensely vascularized zone. IP measurement in this zone was adjusted to the zero level. Diameter of the peripheral zone of vital cells measured 1.3 mm. Capillaries were smaller and sparse. Dynamic MRI revealed peripheral washout of the contrast agent in this zone. After an initial increase of the signal intensity a hypo‐intense rim was formed within a few minutes. The intermediate region was characterized by islands of vital tumour cells containing 3% capillaries (hot spots). The innermost area, the necrotic zone, took 35% of the total tumour area with less than 0.5% vessels. The IP increased from the periphery to the centre. VEGF and VEGF‐receptor 2 was found in the vessels of the tumour margin and vital tumour cells of the peripheral zone. From this can be concluded that the peripheral washout phenomenon seems to be correlated with elevated interstitial pressure and increased capillary density and therefore may be a reliable sign of malignancy.
Leishmaniasis in Canis familiaris – a Report About Microvascular and Cellular Architecture of the Infected Spleen
Authors: Alexandre‐Pires G. , Gilot T. , Pais D. , Correia M. , Esperança Pina J. A.
Published: November 2005

Leishmaniases are a globally widespread group of parasitic diseases from the group of anthropozoonoses. They are caused by an intracellular protozoan parasite that causes a wide spectrum of diseases in humans and dogs worldwide. In fact, the dog is considered one of the most important reservoir. The spleen is one of the several haematopoietic and immunocompetent organs involved. As this organ is a blood filter, the authors looked for the expression of this infection over the microvascular environment and modifications of the spleen cell population related to immunological responses to this parasitic condition.

Tools:  Scanning electronic microscopy over intact tissue and corrosion casts, transmission electronic microscopy, histology and immunohistochemistry (MAC 387 and CD3).

Results:  Our results show three important modifications concerning the microvascular architecture of the spleen when compared to the normal pattern. A striking scarcity of the sinusoidal system sheath that surrounds the central arteries of the white pulp, a huge development of pulp venules and veins, differentiation of typical features of high endothelial venular cells.

Remarks:  According to our results it seems that independent of the virulence of the parasite involved and the kind of immunity prevalent in a particular host, the spleen develops blood dynamic conditions that allows a sluggish blood flow so that cells involved in immunological processes can proliferate and differentiate and it also contributes to the trapping of lymphocytes to the area through the differentiation of typical characteristics of HEV endothelial cells.
Vasculogenesis by Endothelial Progenitor Cells In Vitro
Authors: Käßmeyer S. , Plendl J.
Published: November 2005

Introduction and Aim:  Vasculogenesis is the de novo formation of blood vessels from embryonic precursor cells. The formation of additional vessels is called angiogenesis, the sprouting from pre‐existing vessels by migration, proliferation and differentiation of endothelial cells. Until recently vasculogenesis was considered to be restricted to embryogenesis, however, evidence suggests that progenitor cells exist in the adult, able to participate in neovascularization. In order to study cellular and molecular control mechanisms without animal testing we have established in featuring the three‐dimensional formation of capillary‐like structures from microvascular endothelial cells. The aim of this study was to identify the specific subpopulation of cells that triggers the primary process of vasculogenesis in vitro.

Material and Methods:  Endothelial cells were isolated from the bovine corpus luteum and examined by phase contrast and transmission electron microscopy. Immunolabelling was done for the most important pro‐angiogenic factor (VEGF) and its endothelium‐specific receptors (VEGF‐R1, ‐R2) as well as for endothelial cell surface markers (CD 31, CD51/61, CD29) and presumptive stem cell marker CD 34. Additionally VEGF and VEGF‐R1, ‐R2 expression was demonstrated by polymerase chain reaction (RT‐PCR).

Results:  The beginning of vessel formation in vitro was initiated by confluence of endothelial cells. A linear and circular side‐by‐side arrangement of cells and the formation of capillary‐like structures with an internal lumen could be observed. Specific clusters of cells were identified. These cells were arranged in a star‐like formation, i.e., one particular round cell in the centre surrounded by several cells with extensions. Immunolocalization of VEGF, VEGF‐R1, ‐R2, CD 31, CD51/61, CD29 and CD34 was positive. VEGF‐R2 and CD 34 were found to be co‐localized in specific cells exclusively.

Conclusions:  Results for the first time reveal the specific subpopulation of endothelial cells that trigger the start of vasculogenesis in vitro. These progenitor cells are characterized by a specific morphology, by the presence of endothelial cell markers and by the expression of stem cell marker in vitro models
Morphological Studies on the Development of the Bronchial Epithelium of the Fetal Camel Lung
Authors: Kamel G. , Mubarak W. , Sayed R.
Published: November 2005

Light microscopic observations revealed that in camel foetuses of 25 mm crown‐to‐rump length (CRL) the primordial tubular system of the prospective lung was formed of several tubules lined by undifferentiated columnar epithelium. Intra‐epithelial neuroendocrine cells were the first elements to be differentiated in the lining epithelium of the primordial tubular system of the prospective lung as early as 25 mm CRL. On reaching 50–67 mm CRL, the primordial tubular system started to differentiate into two systems of primordial tubules, the prospective bronchial or light tubules and the future respiratory or dark tubules. The lining epithelium of the prospective bronchial tubules revealed a clear evidence of ciliogenesis as early as 80 mm CRL. From 800 mm CRL onwards, the bronchial epithelium demonstrated ciliated and non‐ciliated secretory cells. The non‐ciliated secretory cells of the bronchial epithelium of fetal camel lung showed moderate reaction to AB/PAS technique, for the first time, in fetuses reaching 600 mm CRL.
Epithelium of the Bursa of Fabricius Mucosa
Authors: Khomych V. , Kalynovska I. , Kolych N. , Mazurkevych T.
Published: November 2005

The structure and topography of the Bursa of Fabricius (BF) differs from other central immune organs. Structural components, providing BF function are located in mucosa, which is constantly influenced by antigen stimulation. Therefore the reports, which have appeared recently, indicate that BF may function as a peripheral immune organ. The latter is known to be situated in the alimentary canal mucosa (caecal tonsils, Peyer's patches) and possess peculiar epithelium. The aim of our study was to investigate the structure and cell composition of BF epithelium in chicken, turkey, quails, ducks and pigeons using light and electronic microscopy. The studies have shown that epithelium overlying BF folds is pseudostratified columnar, but between them and in their base it might be cuboidal or columnar. Pseudostratified epithelium consists of columnar cells with a few microvilli on their apical surface, goblet cells, infrequent endocrine cells and macrophages. BF epithelium is spongious due to its infiltration by lymphoid cells. The large quantity of the latter is found in subepithelial connective tissue. Infiltration by lymphoid cells is considered to be typical for the alimentary tract mucosa, where the aggregated lymphoid nodules are situated. Thus, our study supports the thoughts that BF may have function of central and peripheral immune organ.
Morphometric and Structural Examination of the Collateral Ligaments of the Canine Elbow Joint
Authors: Koch R. , Hemmes M. J. , Meyer W. , Waibl H.
Published: November 2005

The aim of this study was to describe morphological differences between the lateral collateral ligament (LCL) and the medial collateral ligament (MCL) of the canine elbow joint. Forty forelimbs of 22 dogs (adult German shepherd dogs and shepherd mongrels) were dissected. The length and width of defined segments of the ligaments (i. e. total length, areas of origin and insertion, ‘free part’, ‘humeral part’, ‘antebrachial part’) were measured in extension (160°) and flexion (30°). Statistical analyses of the data were performed using the data analysis software SAS 8.0. In addition the collateral ligaments of ten forelimbs of five shepherd mongrels were studied histologically. The LCL differs from the MCL in the following morphological items: a) the areas of origin and insertion are larger and in consequence the ‘free part’ of the lateral ligament is shorter; b) the caudal crus of the LCL is fan‐shaped with a broad insertion area at the ulna whereas the caudal crus of the MCL remains slim; c) in the LCL the ‘humeral part’ varies in length depending on extended or flexed position of the joint, which is due to the spiral shape of the capitulum humeri; d) the microscopical structure of the LCL is organized less tightly; the collagen fibre bundles cross in varying angles. Referring to these details the LCL seems to allow and to limit a slight rotation of the forearm when the elbow joint is extremely flexed.
Functional Morphology of Equine Pre‐ovulatory Cumulus‐oocyte Complexes
Authors: Kölle S. , Duchamp G. , Lahuec C. , Caillaud M. , Sinowatz F. , Goudet G.
Published: November 2005

Knowledge of the functional morphology of equine cumulus‐oocyte‐complexes (COCs) shortly before ovulation is scarce. Therefore the aim of our studies was to characterize COCs matured in vivo by light microscopy, transmission electron microscopy (TEM), scanning electron microscopy (SEM) and immunohistochemistry (ICC). Pre‐ovulatory COCs were obtained by ultrasound‐guided transvaginal aspiration of pre‐ovulatory follicles of 26 pony mares. The morphology of pre‐ovulatory COCs was compared to that of immature COCs obtained by slicing of ovaries from the slaughterhouse. As shown by light microscopy and SEM, immature COCs possess numerous round, densely packed cumulus cells, which contact each other and possess merely single short processes. Contrary, in pre‐ovulatory oocytes the cumulus cells are widely separated but send long cytoplasmic processes to the zona pellucida (ZP). All the cumulus cells are embedded in a huge amount of homogenous extracellular matrix. As shown by alcian blue staining and Periodic Acid Schiff Reaction (PAS) with and without amylase digestion, this matrix contains glycoproteins and acidic and sulphated glycosaminoglycans. In pre‐ovulatory COCs both the oocyte and the cumulus cells produce glycosaminoglycans, whereas immature COCs are negative for alcian blue. Similarly, glycoproteins are synthesized in pre‐ovulatory, but not in immature COCs. As shown by ICC, hyaluronic acid is one of the most abundant mucopolysaccharide in the pre‐ovulatory COC. The high synthetic activity of the cumulus cells in the pre‐ovulatory COC is confirmed by TEM showing densely packed endoplasmic reticulum (ER) and accumulation of glycogen in the cumulus cells. Our results imply that in the equine in vitro maturation of the oocyte is characterized by the cumulus cells synthesizing an extracellular matrix of glycoproteins and acidic and sulphated glycosaminoglycans. The extensive production of extracellular, water‐binding matrix in the pre‐ovulatory COC ensures mechanical protection and nutrition of the oocyte.
Regulation of Cumulus Cell Numbers in the Course of the Canine Estrous Cycle
Authors: Kölle S. , Woehl‐Wenigerkind A. , Sinowatz F. , Braun J.
Published: November 2005

In the canine, success rates of assisted reproduction techniques are very low. Since cumulus cells are known to play an essential role during the in vitro maturation (IVM) of oocytes, the aim of this study was to investigate the regulation of cumulus cell numbers in the course of the canine estrous cycle. Eighty‐two cumulus‐oocyte‐complexes (COCs) were obtained from 16 bitches undergoing elective ovariectomy during proestrus, estrus, metestrus and anestrus. First, mean cumulus cell numbers during the different stages of the cycle were compared. Then the incidence of programmed cell death (apoptosis) was detected by terminal deoxynucleotidyl transferase mediated dUTP labelling (TUNEL). Scanning electron microscopy (SEM) was used to confirm the presence of apoptotic cumulus cells. Additionally, mitosis was detected by using antibodies directed against the nuclear cell proliferation‐associated antigen Ki‐67. The mean number of cumulus cells in the COCs increased from proestrus to estrus (168.2 ± 9.2 to 265.5 ± 32.7, P < 0.01) and decreased from estrus to metestrus (265.5 ± 32.7 to 139.1 ± 10.7, P < 0.01) till anestrus (150.0 ± 8.3, P < 0.01). The percentage of apoptotic cumulus cells was generally very low during proestrus and oestrus (3.3% and 12.6%, P < 0.01) and distinctly increased during metestrus (83.1 %, P < 0.001). The predominance of apoptosis in the cumulus cells during metestrus was confirmed by SEM, which showed a high number of cells revealing cell shrinkage, loss of membrane integrity and cell disruption. Mitosis was highest during proestrus (2.7%) and oestrus (2.6 %) and decreased in the course of metestrus (1.8%) and anestrus (0.2%, P < 0.05).

Our results imply that the distinct increase of cumulus cell numbers during proestrus and oestrus is due to increased mitosis and reduced apoptosis. The distinct reduction of cumulus cells during metestrus is predominantly regulated by apoptosis.
Embryo‐maternal Communication during the First Days of Embryonic Life
Authors: Kölle S. , Wolf E. , Sinowatz F.
Published: November 2005

The mechanisms of embryo‐maternal communication during the first days of embryonic life are largely unknown. Using the bovine as a model, the aims of our study were to morphologically characterize the interaction between the pre‐implantation embryo and the epithelium of the maternal ampulla, isthmus and uterotubal junction by light and scanning electron microscopy. For this purpose, oviducts were removed from cows revealing a functional corpus luteum on day 3 after insemination. These were compared to oviducts removed on day 3 (metestrus) of the estrous cycle. Three days after insemination, the majority of the epithelial cells in the ampulla were secretory cells distinctly protruding into the oviductal lumen. Contrary the ampulla of cows on day 3 of the cycle predominantly revealed ciliated cells in the oviductal epithelium. As shown by Periodic Acid Schiff reaction (PAS) with and without amylase digestion, the secretory cells of the ampulla synthesized merely glycoproteins during metestrus, but large amounts of glycogen during pregnancy. In the isthmus no morphological differences were seen between pregnant and cyclic cows. The most conspicuous finding during pregnancy was seen in the uterotubal junction: Vital cumulus cells embedded in between epithelial cells had developed short cytoplasmic processes intensely contacting the epithelial uterine cells. The embryos obtained ex vivo were regularly covered with a thick layer of homogenous extracellular matrix. Contrary embryos produced in vitro– both with and without coculture with oviductal cells –revealed a clearly visible zona pellucida with spongy appearance and numerous pores. Our results imply that already during the first days of life there is intense interaction of the pre‐implantation embryo and the maternal genital tract part of which may be mediated by cumulus cells.
Laser Micro‐dissection and Quantitative RT‐PCR as a Tool for Cell Identification
Authors: Koppang E. O. , Huang Q. , Hordvik I. , Landsverk T. , Press C. McL. , Bakke‐McKellep A. M.
Published: November 2005

We have applied a laser micro‐dissection microscope followed by RT‐PCR analysis of micro‐dissected tissue to identify gene expression of various immune genes in the Atlantic salmon, Salmo salar. A major problem in teleost anatomy is the lack of specific cell markers to identify different leukocyte populations. For instance, the involvement of T‐cells in inflammatory reactions is unknown. We used frozen sections from the gut of salmon fed to induce localized inflammatory reactions and micro‐dissected regions showing proliferative changes. RNA was extracted from the micro‐dissected specimens and subjected to quantitative RT‐PCR using primers and TaqMan probes for T‐cell receptor, MHC class II and elongation factor, which served as a house‐keeping gene. The results showed expression of T‐cell receptor, MHC class II and elongation factor. The expression of the elongation factor corresponded invariably to the area micro‐dissected tissue. In conclusion, T‐cells are present in teleost inflammatory reactions.
Incompleteness of the Structure of Liver Veins and Tissue Components in Neonatal Animals
Authors: Krishtoforova B. V. , Lemeschenko V. V.
Published: November 2005

Introduction:  The formation of the liver structure at mammal embryogenesis is interconnected by drawing up of venous mains. In parallel to the transformation of the main veins, there is a reduction of the blood formation function and the liver transformation from the primary organ of embryogenesis into the secondary one, but the exception is immaturity and underdeveloped maturity mammals (Smirnova, 1967; Mechtiev, 1968; Bresler, 1993). The purpose of research is to establish morphofunctional criteria of the incompleteness in the blood vessels and in tissue components of the liver in neonatal animals.

Material and Methods:  The afferent and efferent liver veins of calves, piglets and puppies at one day of age were investigated. The complex of morphological, roentgenological, ultrasound and statistic methods was used.

Results:  It was established that the liver veins in newborn animals portoumbilical (afferent) and caval (efferent) collectors. In 1‐day‐old calves, the afferent collector is connected with caval way of the large vein flow of the main type. Relative weight of the liver in the calves is 2.31–2.85%, hepatocytes form balks and they are not always oriented radially to the central venuels. There are no seats of haemopoesis. Sonography of the liver and its veins proves inconstancy of the sizes depending on acts of breath. In 1‐day‐old piglets, many treelike anastomoses follow with the left hepatic vein. Relative weight of the liver in the piglets is 2.27–4.64%. The parenchyma of the organ is spongy with the seats of haemopoesis (0.04 3.61%). In 1‐day‐old puppies afferent and efferent collectors combine through the venous duct. Additionally, in the abdominal cavity of puppies, there are the vitelline artery (starts from a. mesenterica) and vein (runs to v. porta near the afferent collector), following to the umbilical ring. Relative weight of the liver in the puppies is about 5.18–6.04%. Hepatocytes form the chaotic congestion with haemopoetic points (2.04–7.14%).

Conclusion:  Thus, incompleteness of morphology of the liver is shown in preservation of provisional vein structures and tissue components of the embryonic type, which is one of the factors complicating diseases of digestive organs in neonatal animals.
Comparative Morphology of the Arterial Circle of the Brain in Some Species of Mammals
Authors: Kuchinka J. , Nowak E. , Szczurkowski A. , Kuder T.
Published: November 2005

Investigations were performed on 10 adult individuals of chinchilla (Chinchilla laniger), midday gerbil (Gerbillus gerbillus), rabbit (Oryctolagus cuniculus), guinea pig (Cavia porcellus) and Egyptian spiny mouse (Acomys cahirinus). The animals were killed with ether anaesthesia and injected with coloured acryl latex through the abdominal aorta or left heart ventricle. The heads of the animals were fixed in formalin and after decalcification in 5% HNO3, injected arteries of the base of the brain were exposed. The different types of arterial circle of the brain in investigated species were observed. The open type with morphological asymmetry in the anterior part was found in chinchilla. It depended on an unusually strongly developed anterior cerebral artery at the right side. In the midday gerbil the most common form was the closed type of investigated circle, with a lack of typical communicating artery posterior. The morphological asymmetry of the anterior part of the circle without fully developed anterior cerebral artery was found in the rabbit. Typical, the closed type of investigated structure with little individual variability was observed in guinea pig. The most individual variability connected with anterior and posterior part of the Willi's circle was found in the Egyptian spiny mouse where it was difficult to describe the typical arterial circle of the base of the brain.
Veterinary Anatomy Education in Korea
Authors: Lee H. S.
Published: November 2005

In Korea, there are 10 veterinary schools and about 500 new veterinarians are produced annually. Veterinary education is 6‐year‐system composed of pre‐veterinary (2 yrs) and veterinary course (4 yrs). Total of 28 to 33 subjects are taught in professional curriculum, including 21 of 23 subjects recommended by the World Veterinary Association (WVA) as minimum requirements for veterinary education. Only economics and botany are excluded. Currently, 6648 veterinarians are active in the profession in Korea; about 31% of these in clinical practice and 69% in non‐clinical areas. Korea has 3.5 million pets, 10.4 million livestocks, 105 million chickens, and 0.5 million other animals. In general, 630 livestock units (LU) are covered by each veterinarian. With improvement of economic status and life style change, the number and variety of species of companion animals are expanding, accordingly. Also the importance of molecular biology is recognized and included in the professional curriculum. In these changing times, it is believed that gross anatomy, histology and embryology are essential integral component of found in veterinary education. But the teaching hours of histology, embryology and gross anatomy have reduced 15% during last 5 years due to new subjects. Total teaching hours in professional curriculum is about 4100 hours (pre‐clinical 29%, paraclinical 32%, clinical course 38%). Average 200 hours are allocated for gross anatomy, histology and embryology lecture and average 250 hours for laboratory practice. The number of students per class ranges 40–80 depending on schools. The ratio of male and female students is 6–4, and the teaching faculty and students 1–22. The Korean Association of Veterinary Anatomists (KAVA) recommend one cow, five dogs, two pigs, five goats and 20 chickens per 20 students to be used for the dissection. The students are evaluated using various types of exam such as assignments, written and oral tests. Recently some schools are beginning to adopt block lectures and/or PBL.
Calcium‐Binding Proteins and Neurotransmitters in the Stomach Myenteric Plexus of the Korean Native Goat
Authors: Lee H. S. , Lee I. S. , Nam Y. S. , Yoon Y. S.
Published: November 2005

The expression of calbindin D‐28k (CB), calretinin (CR), substance P (SP) and calcitonin gene‐related peptide (CGRP) in the stomach myenteric plexus of the Korean native goat stomach was investigated by immunohistochemistry. The results demonstrated the presence of nerve fibers and cell bodies immunoreactive (IR) to CB, CR, SP and CGRP. In tissues of rumen, reticulum, omasum and abomasum, some distinct neuronal populations could be distinguished according to their morphologic and neuronal chemical properties: Dogiel type I cells which have irregular lamellar dendrites and a single axon, Dogiel type II cells which have large ovoid cell bodies and several long axon‐like processes, and small filamentous interneurons. CB‐, CR‐, SP‐ and CGRP‐IR neurons and fibers were observed in the myenteric plexus of stomach, and varicose nerve fiber immunostained to SP and CGRP also were found in the muscle layer. In myenteric plexus of the stomach, CB‐ and SP‐positive neurons were characterized by Dogiel type II and CR‐IR neurons were classified Dogiel type I with lamellar dendrites, and immunoreactivity of CGRP was very weak in the somata. SP‐ and CGRP‐IR nerve fibers formed dense networks within the myenteric ganglia. SP‐IR cell bodies and their fibers were found in the myenteric plexus, and the immunoreactivity and number of cell bodies were more than CB‐, CR‐, and CGRP‐IR neurons. These results suggest that SP, CGRP, CB and CR in the myenteric neurons of Korean native goat stomach may have play an important role in the dynamic movement.

(Support contributed by: Korean Research Foundation 2003‐015‐E00195).
Effect of Flooring Systems on Microarchitecture of the Bovine Hoof Horn
Authors: Lendner G. , Mülling Ch. , Nebel U. , Bergsten Ch. , Magnusson Ch.
Published: November 2005

Introduction and aims:  Claw diseases in dairy cows not only lead to high economic losses, but also cause pain and reduce animal welfare. Former studies pointed out that flooring properties influence the microstructure of hoof horn. In this study, we looked for correlations between different flooring systems and horn quality. We obtained horn samples from an experimental farm in Alnarp/Sweden from cows that were first kept on slatted floor and subsequently on mastic asphalt respectively on rubber mats.

Methods:  Untreated horn samples were glued onto wooden blocks: 7 μm sections were prepared with a Polycut‐S (Leica) microtome and afterwards stained with the PAS reaction. For morphometrical analysis we used a Zeiss Axioskope, a digital camera (Progress 3012) and the Lucia 32 G software (Nikon). These data were exported into EXCEL (Microsoft) and statistical analysis was carried out with SPSS (SPSS Inc. [[Au: please provide manufacturer details for ‘SPSS’: city, state code (if USA) and country]]). We measured area, minimal and maximal diameter of medulla and cortex of the horn tubules. Statistical evaluation included number of tubules and tubular area per visual field, total area of medulla and cortex and the limiting diameters.

Results:  We found significant alterations of the hoof horn after moving the cows from hard to soft floor. The diameter of the horn tubules decreased significantly. The area of medulla and the ratio of medulla to cortex decreased as well.

Conclusions:  Our findings indicate a higher quality of claw horn and an increased resistance against mechanical, chemical and microbial environmental influences in animals kept on soft floors. The functional implications of these observations are discussed with regard to claw health as well as the arising consequences for dairy housing systems.

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.