In vitro studies of a photo-oxidized bovine articular cartilage     

Akens MK  von RB  Bittmann P  Nadler D  Zlinszky K  Auer JA 《Journal of veterinary medicine. A, Physiology, pathology, clinical medicine》2002,49(1):39-45

Bovine articular cartilage was photo-oxidized and cultured with native articular bovine cartilage and synovial membrane to study the interaction between these tissues mimicking the physiological situation in the joint. The photo-oxidation was applied as a pretreatment of cartilage for future use in cartilage resurfacing procedures in joints. Properties of the transplant were assessed by testing the production of local mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), and neutral metalloproteinase activities under normal conditions and after stimulation with various stimulants representative of inflammatory changes in pathophysiological conditions. Unlike normal cartilage photo-oxidized cartilage did not release significant amounts of NO and PGE2 and showed less gelatinolytic and caseinolytic activity compared to native bovine articular cartilage. Enzyme activity of the combined cultures was at a level intermediate between that of photo-oxidized cartilage and native cartilage cultures alone. In contrast to normal cartilage, living chondrocytes were not visible in photo-oxidized cartilage using live/dead staining. These results indicate, that the photo-oxidized cartilage may have a beneficial effect on adjacent native host cartilage and therefore be a suitable transplant for use in in vivo experiments.  相似文献   

Evidence for oxidative stress involved in physiological leaf spot formation in winter and spring barley     

Wu YX  von Tiedemann A 《Phytopathology》2002,92(2):145-155

ABSTRACT A leaf spot disease with unknown etiology has become more pronounced in spring and winter barley in Germany in recent years. The symptoms are similar to net blotch and Ramularia leaf spots, but the causal agents of these diseases are not identified. The symptom expression varied much on cultivars. Cultivars most affected by the disease of both spring and winter barley showed a significantly higher level of superoxide (O(2) ) production and lipid peroxidation (malondialdehyde), but a lower level of antioxidant potential expressed as superoxide dismutase (SOD) activity, catalase activity, and integral water-soluble antioxidant capacity (ACW) than insensitive cultivars. A high positive correlation between O(2) production and leaf spot development between ear emergence and milk ripeness was established in the most sensitive winter barley cv. Anoa (r(2) = 0.9622) and spring barley cv. Barke (r(2) = 0.9434). Leaf H(2)O(2) levels increased with the severity of leaf spots. The histochemical localization of O(2) and H(2)O(2) in the tissues adjacent to leaf spots indicated that these two active oxygen species (AOS) are involved in the formation of leaf spots. Reduction of symptom severity by applying strobilurin and azole fungicides was always associated with elevated SOD activity and ACW content and suppressed O(2) production. However, peroxidase activities were significantly higher in sensitive cultivars and in more severely affected tissues and decreased by applying fungicides. Thus, it is assumed that a possible genetic mechanism based on the imbalanced AOS metabolism contributes to formation of physiological leaf spots.  相似文献   

Results of the German investigation on the EU project "Salmonella in Pork (Salinpork)"--1. Investigations in the farms     

von Altrock A  Schütte A  Hildebrandt G 《Berliner und Münchener tier?rztliche Wochenschrift》2000,113(5):191-201

Reports of human salmonellosis caused by the consumption of pork and the introduction of control and surveillance programmes in different countries of the European Union were the reason for carrying out an international study under the title "Salmonella in Pork (Salinpork)": Six different EU-countries were involved in this study, which was supported by the Commission of the European Community and which was carried out over a period of April 1996 to April 1999. The aim of the investigation presented was to determine the prevalence of Salmonella in fattening, breeding and farrow-to-feeder herds as well as the determination of risk factors for the introduction of Salmonella into the farm (Part 1). In addition, sources of contamination of pork should be detected by taking samples of the product and the environment in the slaughterhouse (Part 2). In Germany, the investigation into Salmonella infections of 60 fattening, 20 breeding and 20 farrow-to-feeder herds were carried out in Schleswig-Holstein. The investigation included bacteriological examinations of feed and faecal samples for sero- and phagetyping and serological examinations by using the Danish Mix-ELISA. From 2,947 serological investigated fattening pigs were 7.3% (n = 213) positive, from 797 breeding sows 9.2% (n = 73) were serological positive and 4.5% (n = 18) of the investigated sows (N = 399) in farrow-to-feeder herds were serological positive. Altogether, 28.3% of the fattening, 50.0% of the breeding and 15.0% of the farrow-to-feeder herds were serological positive. A questionnaire was used to capture data about management, hygiene measures, feeding systems and the occurrence of diseases in the herd. After statistical analysis the common risk factor of fattening herds and sow herds was the use of pelleted feed. But in a control study with 17 different fattening herds the result could not be proven. Other factors which can influence the occurrence of Salmonella infections were discussed.  相似文献   

Results of the German investigation in the EU Project "Salmonella in Pork (Salinpork)"--2. Investigations in a slaughterhouse     

von Altrock A  Schütte A  Hildebrandt G 《Berliner und Münchener tier?rztliche Wochenschrift》2000,113(6):225-233

In 1997 bacteriological examinations for the distribution of Salmonella in slaughterhouses were carried out in Germany within the framework of an international study "Salmonella in Pork (Salinpork)". During 6 days, 1,200 swab and water samples from slaughtered pigs and the environment were taken. 4.4% of the samples (n = 53) were Salmonella positive. S. typhimurium was isolated mainly (69.8%; n = 37), and 6 phagetypes were differentiated. In addition, S. derby and S. panama could be demonstrated. The resistance pattern of the different isolated S. typhimurium-phagetypes are presented. The phagetype DT 104 was multiresistant to ampicillin, spectinomycin, streptomycin, sulphonamide and tetracycline. In comparison with the serological prevalence of 7.3% of the fattening pigs in the farms (Part 1), only 1.0% of the samples taken from the surface of the carcass were Salmonella-positive. Swabs taken from the liver were in 2.7% positive and samples from the tongue gave in 5.3% of the cases Salmonella-positive results. In the examination of the environment Salmonella was demonstrated mainly from the water outlets, whereas Salmonella could not be isolated from water of the scalding tank. There was only one case (0.7%) in which Salmonella could be isolated from the hands of the personnel, and also only one swab of the polishing machine was positive (1.1%). But 6.7% samples of the saw were Salmonella-positive. A comparison of repeated, at intervals taken samples showed that the number of Salmonella-positive samples was higher in the last examination round of the particular slaughter days. The reason is suspected in the increasing number of slaughtered pigs and supplying farms, which may increase the probability of bringing in Salmonella.  相似文献   

A Universal `One-tube' RT-PCR Protocol for Amplifying Isolates of Bovine Viral Diarrhoea Virus     

Pfeffer M  Freyburg MV  Kaaden OR  Beer M 《Veterinary research communications》2000,24(7):491-503

The increase in the knowledge of the genetic variability of BVDV and the identification of some of the genetic determinants of its pathogenicity require robust and practical tools for rapid molecular characterization of the various genotypes of this virus. This study was undertaken to develop a standard protocol for RT-PCR that allows the amplification of various parts of the genome of BVDV without the need for optimizing each individual reaction. The reaction set-up is very flexible because it consists of two pre-mixes. These are a master mix, with all the required reagents except the desired primers, which are the components of the second pre-mix and are therefore easily interchangeable between the different reactions. After adding any primer-containing pre-mix to the fixed master mix, a non-interrupted cycling protocol led to the generation of amplicons of up to 4 kbp in size in amounts sufficient for subsequent sequencing reactions. The method was applied to five different regions of the BVDV genome: (i) the well-known 5-UTR to differentiate genotypes I and II; (ii) the entire E2 gene, or an approximately 550 bp region within the E2 gene, in order to find the molecular equivalent of antigenic varieties; (iii) the entire structural protein coding region covering the Npro, capsid, E ^RNS, E1 and E2 genes; (iv) a 2.1 kbp region embracing the NS2/3 junction which is known to be cleaved in cytopathic biotypes of BVDV; and (v) the region covering the entire NS4B and NS5A/B genes. All six RT-PCRs were successfully applied using (i) primers with lengths of between 20 and 52 nucleotides, (ii) an aliquot of RNA extracted from either 10⁶ infected bovine embryonal lung cells or the same number of leukocytes from viraemic cattle, and (iii) all the genotype I and II strains of BVDV tested. The technique described was used to generate various Sindbis virus/BVDV recombinants. The correct processing of the amplicon-derived E2 glycoprotein of BVDV strain PT810 was demonstrated by its reaction with a monoclonal antibody in an immunofluorescence assay. Given the variety of RT-PCRs tested, we conclude that this universal protocol may be useful with other RNA viruses.  相似文献   

Spontaneous production of nitric oxide (NO), prostaglandin (PGE2) and neutral metalloproteinases (NMPs) in media of explant cultures of equine synovial membrane and articular cartilage from normal and osteoarthritic joints     

von Rechenberg B  McIlwraith CW  Akens MK  Frisbie DD  Leutenegger C  Auer JA 《Equine veterinary journal》2000,32(2):140-150

Nitric oxide (NO), prostaglandin E2 (PGE2), and the activity of neutral metalloproteinases (NMPs) were measured in conditioned media of equine synovial membrane and articular cartilage explant cultures from horses with normal joints (n = 7) and from horses affected with moderate (n = 7) or severe osteoarthritis (n = 14) as judged by macroscopic appearance. Normal articular cartilage appeared glossy and bluish-white, was of normal thickness and showed no evidence of discolouration, fibrillation or other cartilage discontinuity. Slight discolouration and fibrillation or minor clefts of the cartilage were considered as moderate OA, whereas erosions of articular cartilage down to the subchondral bone were considered as cases of severe OA. Explant cultures of equine synovial membrane and articular cartilage released the local mediators, NO and PGE2, as well as detectable levels of NMP activity into culture media. Concentrations of NO were higher in articular cartilage explants compared to synovial membrane explants, whereas concentrations of PGE2 were higher in synovial membrane explants. The NMPs with collagenolytic activities were similar in both explant cultures, whereas gelatinolytic activities were higher in synovial membrane explant cultures and caseinolytic activities were generally higher in articular cartilage explant cultures. Furthermore it was shown that concentrations or enzyme activities increased according to the severity of disease of the joints. Concentrations for NO, collagenolytic and gelatinolytic NMPs were relatively stable, whereas PGE2 and caseinolytic NMP concentrations increased over time in culture.  相似文献   

Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs   总被引：5，自引：0，他引：5  

Erika Varkonyi-Gasic  Rongmei Wu  Marion Wood  Eric F Walton  Roger P Hellens 《Plant methods》2007,3(1):12

MicroRNAs (miRNAs) are a class of small non-coding RNAs with a critical role in development and environmental responses. Efficient and reliable detection of miRNAs is an essential step towards understanding their roles in specific cells and tissues. However, gel-based assays currently used to detect miRNAs are very limited in terms of throughput, sensitivity and specificity. Here we provide protocols for detection and quantification of miRNAs by RT-PCR. We describe an end-point and real-time looped RT-PCR procedure and demonstrate detection of miRNAs from as little as 20 pg of plant tissue total RNA and from total RNA isolated from as little as 0.1 μl of phloem sap. In addition, we have developed an alternative real-time PCR assay that can further improve specificity when detecting low abundant miRNAs. Using this assay, we have demonstrated that miRNAs are differentially expressed in the phloem sap and the surrounding vascular tissue. This method enables fast, sensitive and specific miRNA expression profiling and is suitable for facilitation of high-throughput detection and quantification of miRNA expression.  相似文献   

THE PHYSIOLOGIC RESPONSE OF DOMESTIC ANIMALS TO IONIZING RADIATION: A REVIEW     

Christina  von ZallingerDVM  Karlheinz  TempelDVM 《Veterinary radiology & ultrasound》1998,39(6):495-503

With respect to clinical signs of the radiation syndromes, some remarkable species variations exist. For example the marked delayed reaction of the acute hematologic response in cows. An unusually high sensitivity of the central nervous system is found in burros, which is probably caused by acute vascular and/or metabolic changes in the brain. The species-specific number of intestinal crypt and hemopoietic stem cells may explain the early survival differences among species after high doses of irradiation. Mortality due to acute radiation syndromes is lowest in chickens. Regarding late effects, various neoplasms are typical in dogs, and cattle more commonly develop cataracts.  相似文献   

Comparative serological studies of methods for demonstrating antibodies in infectious laryngotracheitis of chickens     

B Fuchs  G B?ddecker  V von Bülow 《Berliner und Münchener tier?rztliche Wochenschrift》1985,98(8):261-266

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Die Entwicklung holzverkohlender Betriebe     

H. D. von Bonin-Poniß 《Forstwissenschaftliches Centralblatt》1929,51(14):509-521

Ohne Zusammenfassung  相似文献