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Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold‐free three‐dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19‐day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14‐ and 19‐day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki‐67 immunoreactivity showed an even distribution in two‐dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis‐associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold‐free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell–cell and cell–matrix interactions.  相似文献   
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A field study was conducted aimed at (i) evaluating the practicability of a fixed‐time insemination regime for medium‐sized dairy operations of north‐western Germany, representative for many regions of Central Europe and (ii) substituting hCG for GnRH as ovulation‐inducing agent at the end of a presynch or ovsynch protocol in an attempt to reduce the incidence of premature luteal regression. Cows of two herds synchronized by presynch and two herds synchronized by ovsynch protocol were randomly allotted to three subgroups; in one group ovulation was induced by the GnRH analog buserelin, in another by hCG, whereas a third group remained untreated. The synchronized groups were fixed‐time inseminated; the untreated group bred to observed oestrus. Relative to untreated herd mates, pregnancy rate in cows subjected to a presynch protocol with buserelin as ovulation‐inducing agent was 74%; for hCG it was 60%. In cows subjected to an ovsynch protocol, the corresponding relative pregnancy rates reached 138% in the case of buserelin and 95% in the case of hCG. Average service interval was shortened by 1 week in the presynch and delayed by 2 weeks in the ovsynch group. It may be concluded that fixed‐time insemination of cows synchronized via ovsynch protocol with buserelin as ovulation‐inducing agent is practicable and may help improve efficiency and reduce the work load involved with herd management in medium‐sized dairy operations. The substitution of hCG for buserelin was found to be not advisable.  相似文献   
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OBJECTIVE: To determine historical, physical, and microbiologic findings in horses with limb cellulitis and identify factors associated with short- and long-term outcome. DESIGN: Retrospective case series. ANIMALS: 44 horses with limb cellulitis. PROCEDURES: Information obtained from medical records included use, history, affected limb, diagnostic procedures, treatment, and short-term outcome. Long-term follow-up information was obtained by means of a telephone survey. RESULTS: Twenty-four horses had primary cellulitis, and 18 of the 24 (75%) had a hind limb affected. Results of microbiologic culture were positive for all 20 horses with primary cellulitis from which specimens were obtained, with coagulase-positive Staphylococcus spp recovered from 12 of the 20. Short-term survival rate was 67% (16/24) for horses with primary cellulitis; 7 of the 9 horses available for long-term follow-up were being used for their intended use, and 4 had had a recurrence. Results of microbiologic culture were positive for 13 of the 16 (81%) horses with secondary cellulitis from which specimens were obtained. Short-term survival rate for horses with secondary cellulitis was 90% (18/20). Eleven of the 17 horses available for long-term follow-up were being used for their intended use; 2 had had a recurrence. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that cellulitis can be a life-threatening condition in horses. Horses that were febrile at admission or that developed laminitis were significantly less likely to survive. The prognosis for return to function was guarded, and recurrence was a potential concern.  相似文献   
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Between 2000 and 2004 a disease occurred in an aviary in Germany affecting various bird species belonging to the order Passeriformes including Collared Grosbeaks (Mycerobas affinis), Eurasian Bullfinches (Pyrrhula pyrrhula griseiventris), Brown Bullfinches (Pyrrhula nipalensis), Grey-headed bullfinches (Pyrrhula erythaca) and Yellow-bellied Tits (Periparus venustulus).The major clinical signs included increased mortality of fledglings and young birds, as well as feather disorders and feather loss in adult birds. In addition, adult Eurasian Bullfinches showed in one year a disease course, in which the major symptom was inflammation of the skin beginning on the basis of the beak and spreading over the head occurring a few days before death. Bacteriological and parasitological investigations did not reveal any consistent findings. Using a newly developed polymerase chain reaction protocol, DNA of the recently discovered finch polyomavirus (FPyV) was demonstrated in several affected birds. Because of the consistent detection of FPyV-DNA and the similarity of the symptoms with those observed during infection with the closely related avian polyomavirus in other bird species, an etiological role of FPyV in the observed disease is assumed.  相似文献   
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To determine if Moraxella bovoculi (M. bovoculi), a recently characterized coccoid Moraxella that was isolated from the eyes of calves affected with infectious bovine keratoconjunctivitis (IBK), and Moraxella ovis (M. ovis), originally isolated from sheep with conjunctivitis, possessed genes encoding RTX proteins, genomic DNA was amplified with oligonucleotide primers targeting RTX operon genes of Moraxella bovis (M. bovis). Complete classical RTX operons composed of RTXCABD genes closely linked to a putative secretion accessory protein encoding gene (tolC) were identified in M. bovoculi and M. ovis and were designated mbvCABDtolC and movCABDtolC, respectively. These genes were closely related to M. bovis mbxCABDtolC. Polyclonal rabbit antiserum against the carboxy terminus of M. bovoculi MbvA neutralized hemolytic activity of both M. bovoculi and M. ovis; this antiserum did not neutralize the hemolytic activity of M. bovis. M. bovoculi and M. ovis possess genes that encode proteins related to pathogenic factors of M. bovis.  相似文献   
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