Gregor Majdič and young researcher Katerina Čeh have developed a new method for healing arthritis and other joint problems in dog. As far as we know, Gregor Majdič's research group is the third in the world and second in Europe that will offer such treatment commercially to dog owners. This is a new, highly technological medical method (developed purely by our own knowledge) with potentially huge market. Therefore, we established a new spin-off company called Animacel, ltd at the beginning of 2011 for commercialization of this method. For this innovation, Gregor Majdic was awarded two awards for best innovations in public sector in Slovenia in 2011. During award ceremony for the best innovation at University of Ljubljana, this innovation was described with following words: “Company Animacel with business plan »Regenerative treatment of domestic animals« has pretty much everything that perspective, highly technological company should have – important, even revolutionary innovation, huge potential market, well prepared business strategy, investor and references from the first satisfied patients. The company has even more than that. The company works in the field of stem cells, where the team behind this project has, with its innovative process for use of stem cells, opened very large area for further studies as well as applications, not just in veterinary medicine, where they are currently active, but also in human medicine. We sincerely hope that their contribution will have important impact on health and better quality of life for animals in humans.” Animacel ltd. was established in February 2011. The company provides stem cell treatments for animals, what is completely novel treatment paradigm for joint and ligament problems that were until now incurable. Similar problems in dogs and horses were until now treated only with pain alleviation while our original procedure offers the healing of the pathological process for the first time. According to our experiences so far, with more than 40 dogs already treated, majority of dogs do not need any pain alleviating treatment three months after our procedure. The treatment is therefore also cost effective for the owner since the price is similar to one year supply of pain alleviating medicine. The procedure was developed autonomously in the laboratory of Professor Gregor Majdič at the Veterinary faculty, University of Ljubljana. By the end of 2011, we have treated 40 dogs and 2 horses with different problems with locomotor apparatus (arthritis, joint dysplasia, tendonitis). All patients are still regularly monitored and in all treated dogs and both horses, there was a marked improvement of their health/ability to walk without pain. Animacel ltd. is collaborating with Veterinary hospital Šentjur, Veterinary faculty in Ljubljana, Veterinary hospital Slovenska Bistrica and Veterinary hospital Vetpoint in Zagreb, Croatia. As far as we know, there are currently two other companies in the world that perform stem cell treatments in animals, one based in US and the second based in Belgium. We believe that we have important advantages in both our procedure and market plan. In comparison to the US company, we need much less adipose tissue to perform the treatment, what is especially important in small and very lean dogs, where large amount of tissue, required by US company, is very difficult to obtain. We are not very familiar with the procedure of Belgian company but our advantage is that we are directly present in Slovenian and neighbouring, very large markets (Italy, Austria, Germany), and also our marketing plan which is focused on direct contacts with veterinarians and animal owners. Besides that, the market for stem cell treatments in animals in US, EU and other countries is so large that there is enough marketing potential for all three companies. Furthermore, we have advantage over potential new competitors on the market by already developed and proven protocol for treatment, establ
F.20 Company spin off
Professor Zlatko Pavlica was awarded the FECAVA prize for the best new work paper in the European Journal of Companion Animal Practice (EJCAP) for the year 2011. In 2011 the selection was made among the articles published in the field of veterinary dentistry and the paper "Periodontal disease from the whole body perspective" was chosen for the award. The award was presented to Professor Pavlica in September 2011 at the 17th FECAVA Eurocongress in Istanbul. This award is a reflection of Professor Pavlica committed long-term clinical and research work as well as cutting-edge expertise in the field of periodontal medicine of small animals. Periodontal disease is the most common chronic infection in dogs and humans, affecting tooth supporting tissues and leading to tooth loss. Although periodontal disease is primarily an infectious disease, the disease progression is determined by the critical loss of the balance between bacteria and the host response. The importance of oral focal infection for the systemic health was already mentioned at the end of 19th century, however systemic diseases were only recently associated with periodontitis. The common denominator of many mechanisms that connect oral focal infection with systemic effects is the chronic state of systemic inflammation that accompanies severe periodontitis. Periodontitis is characterized by elevated levels of acute phase proteins such as C-reactive protein (CRP), pro-inflammatory cytokines (interleukin (IL)-6) and coagulation factors (fibrinogen). The same can also be observed in patients with gingivitis only. However, there is a speculation that a common underlying hyper-inflammatory phenotype might predispose animals and humans, to both periodontitis as well as systemic inflammation. These bidirectional effects mean that elevated levels of inflammatory biomarkers may signify both the causes and consequences of periodontitis. Epidemiological studies have shown that patients with periodontitis are at significantly higher risk of developing cardiovascular disease. Gram-negative bacteria or their lipopolysaccharides are reported to induce atherosclerosis-like lesions in experimental animals, and similar data comes from human clinical studies, where traces of periodonto-pathogenic bacteria were detected in atherosclerotic plaques. Although bacteraemia in periodontal disease patients is suggested to be a passive event, some periodonto-pathogenic bacteria can actively spread and enter endothelial cells. Bacterial endocarditis is also reported to be a possible sequel of bacterial spread from the oral cavity. In dogs, periodontal disease has been described in association with atrioventricular valve disease and myocardial disease. Not only bacteria, but also infection-associated increased systemic levels of pro-inflammatory cytokines, and systemic (nitr)oxidative stress in periodontal disease patients are likely involved in the development of cardiovascular disease. When oral bacteria and/or their products/toxins are inhaled aspiration pneumonia can develop. Poor oral health, especially if the periodonto-pathogenic bacterium Porphyromonas gingivalis is present in the oral cavity, has been described to increase risk for aspiration pneumonia. This might be related to the immunomodulatory effect of some periodonto-pathogenic bacteria, leading to increased susceptibility for bacterial tissue colonisation and infection. Our study revealed that aspiration pneumonia caused by inoculation of experimental animals with dead or live P. gingivalis increases systemic pro-inflammatory cytokine levels that may affect distant tissues and organs. However, it seems that aspiration and/or ingestion of P. gingivalis inhibits systemic immune response at the very early phases of infection as determined by systemic nitric oxide levels. Nitric oxide has an important protective role in P. gingivalis infections, its’ reduced levels might indicate a transient tolerance of the host to the presence of P. g
E.02 International awards
Tamara Dolenšek and Tanja Knific, students of the 5th year at Veterinary Faculty received Prešeren award of University of Ljubljana for year 2011 for student's research work: «Separation of apoptotic boar spermatozoa using magnetic activated cell sorting with annexin V-conjugated microbeads«. Work was done under the supervision of Assit. Prof. Petra Zrimšek, PhD, and co-supervision of Assist. Prof. Janko Mrkun, PhD in the frame of the research programme P4-0053 in the area of investigation that contributes towards improvements in the semen preservation biotechnology. Programmed cell death (apoptosis) most likely contributes to failed assisted reproductive techniques and to the decrease in semen quality after cryopreservation. One of the features of apoptosis is the externalization of phosphatidylserine (PS) which could be used to remove apoptotic cells from semen preparations. Magnetic-activated cell sorting (MACS) using annexin V-conjugated microbeads which bind to PS was already successfully used to enhance human semen quality for assisted reproductive techniques, whereas boar spermatozoa were separated by MACS for the first time in the present study. Additionally, we also performed the technique using microbeads that recognize an antigen in the plasma membrane of apoptotic as well as dead cells. MACS of 12 boar semen samples after 3 days of liquid storage at 16 - 17 °C delivers two fractions: unbound (non-apoptotic/live spermatozoa) and bound (apoptotic/dead spermatozoa). Flow cytometry enabled analysis of subpopulations according to membrane integrity. Staining by Annexin V-conjugated with Alexa Fluor 488 (A) and propidium iodide (PI) revealed four subpopulations: live (A-/PI-), early apoptotic; live (A+/P-), late apoptotic and early necrotic; dead (A+/PI+), late necrotic; dead (A-/PI+). Comparing bound fractions obtained by both procedures as well as the unbound ones reveals no significant difference in any of the tested semen parameters. Although early apoptotic and late necrotic (dead) spermatozoa were found in all semen samples, their frequency was significantly higher in bound in comparison to unbound fractions and to control samples. The lowest level of live spermatozoa was found in bound fractions and it differs significantly in comparison to control samples. Motility and progressive motility obtained by computer-assisted semen analysis (CASA) and viability according to Hoechst staining decreased in unbound fractions in comparison to control samples, whereas the results of bound fractions obtained by CASA are irrelevant due to a small number of spermatozoa in samples analyzed. Morphological changes of spermatozoa assessed on Giemsa-stained samples occurred during MACS. An unusual anomaly on the head of spermatozoa was observed in both fractions. A significant decrease in proportion of morphologically normal spermatozoa and an increase in spermatozoa subpopulation with detached acrosome were observed in fractions after MACS in comparison to control samples. DNA fragmentation was detected by introducing a commercial Halomax® test kit. The level of DNA fragmentation showed by spermatozoa heads with large halos of diffused spots did not differ significantly among the control samples and fractions, whereas a significant positive correlation was observed between DNA fragmentation and the presence of cytoplasmic droplets in control samples. MACS for separation of apoptotic and dead cells caused damage to the boar spermatozoa in terms of motility, viability and morphology. Therefore, the results of our study show that MACS seems to be inappropriate for boar semen separation under tested conditions, although it has been successfully used for excluding apoptotic spermatozoa from human semen. Research work enables further studies, also with colleagues from Faculty of Medicine in the field of assisted reproduction technology.
E.01 National awards
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