Marija Lipar

Bone morphogenetic protein (BMP)1-3 enhances bone repair

Members of the astacin family of metalloproteinases such as human bone morphogenetic protein 1 (BMP-1) regulate morphogenesis by processing precursors to mature functional extracellular matrix (ECM) proteins and several growth factors including TGFβ, BMP2, BMP4 and GFD8. We have recently discovered that BMP1-3 isoform of the Bmp-1 gene circulates in the human plasma and is significantly increased in patients with acute bone fracture. We hypothesized that circulating BMP1-3 might have an important role in bone repair and serve as a novel bone biomarker. When administered systemically to rats with a long bone fracture and locally to rabbits with a critical size defect of the ulna, recombinant human BMP1-3 enhanced bone healing. In contrast, neutralization of the endogenous BMP1-3 by a specific polyclonal antibody delayed the bone union. Invitro BMP1-3 increased the expression of collagen type I and osteocalcin in MC3T3-E(1) osteoblast like cells, and enhanced the formation of mineralized bone nodules from bone marrow mesenchymal stem cells. We suggest that BMP1-3 is a novel systemic regulator of bone repair.

Antimicrobial prophylaxis in addition to premedication in gastrointestinal surgery.

Abstract The aim of this study was to evaluate three combinations of antibiotics applied perorally prior to laparotomy and gastrointestinal surgery and to select the most favorable combination. Research was performed on 24 female and male dogs divided into four groups. Following abdominal wall incision the caecum was compressed manually and 5 mL of saline solution was injected into it. Five mL of caecum contents was aspirated in a sterile syringe and only 0.1 mL was incubated into blood agar and homogenous agar. Bacterial colonies were counted and determined. Prior to surgery the control group (n=6) received no antibiotics. The group which received the combination of gentamicin and clindamycin (n=6) achieved the best reduction of E. coli whereas Enterococcus faecals was 100% destroyed. The group which received amoxicillin with clavulanic acid and metronidazole (n=6) acquired a reduction of bacteria Enterococcus faecalis, which could be sufficient for prophylaxis. The combination of gentamicin and erythromycin (n=6) failed to accomplish a reduction of the number of bacteria.

Progesterone and Insulin-Like Growth Factor I Levels in Blood of Boer Goats During Puerperium Out-of-Season in a Mild Climate Region

The aims of this study were to compare insulin-like growth factor-I (IGF-I) and progesterone concentrations as indicators of cycle activity in the blood of Boer goats during puerperium out-of-season in mild climate conditions and to investigate the influence of parity and litter size on the concentration of IGF-I and progesterone in serum during the study period. Twenty Boer (8 primiparous and 12 pluriparous) goats were examined in this study. Blood samples were taken every 4 days starting on the 3rd day and up to the 40th day of puerperium. IGF-I and progesterone (P4) blood levels were determined using radio immune assay (RIA). Results indicated cyclic ovarian activity outside the breeding season in 18 of 20 Boer goats. IGF-I levels in blood sera followed oestrus and cyclic ovarian activity. IGF-I concentrations correlated significantly with P4 concentrations. The IGF-I peak preceded the P4 peak by approximately 8 days in cycling goats. IGF-I and P4 blood levels were not influenced by parity or litter size. Changes in blood serum P4 and IGF-I concentrations levels during puerperal period can assist in the evaluation of reproductive status of goats.

Influence of different therapy approaches - with or without manual extraction - of retained placenta on reproductive performance in simmental cows

The aim of the research was to determine the influence of different methods used in the therapy of retained placenta on conception rate and duration of days open in cows. Research was performed on 241 Simmental cows aged 2-8 years divided in three groups. The first group of cows (group A) (n=82) with retained placenta was treated with manual extraction of placenta 12-24 h after parturition combined with application of intrauterine antibiotics. The second group (group B) (n=79) with retained placenta was treated with intrauterine antibiotics only 12-24 h after parturition and repeated 2-3 times in 48 h intervals. The third group (group C) (n=80) was the control group and had physiological puerperium (without retained placenta). To assess the reproductive performance of Simmental cows, interval from calving to first insemination (days open to first service, DOFS), interval from calving to pregnancy (days open to pregnancy, DOP), relative pregnancy rate (%), first service conception rate (FSCR, %) and all service conception rate (ASCR, %) were measured. The estimate of hazard ratio for DOFS in group A relative to cows in group B was 2.20 (p<0.0001), implying that cows in group B had oestrus earlier and were open shorter until the first insemination, that is, the relative rate of first service decreased by 54.5% in the group A while holding all other variables constant. The variable that had a significant influence on DOFS was lactation (HR=0.99; p<0.0001). The estimate of hazard ratio for DOP in group A relative to cows in group B and C was 3.53 (p<0.0001) and 1.73 (p<0.0001), respectively, implying that cows in group B and C were open longer until pregnancy, that is, the relative rate of pregnancy in group A decreased by 71.6% and 42.2% in comparison with group B and C while holding all other variables constant. Variables that had a significant influence on DOP were lactation (HR=0.99; p<0.0001) and peripartal diseases (HR=0.48; p=0.02). The estimate of hazard ratio for DOFS and DOP between group B and C was not significant. Cows treated with intrauterine foaming tablets after RFM became pregnant earlier and had a better conception rate in contrast to cows treated with manual extraction of placentae.

Canine babesiosis treatment with three different medicines

Canine babesiosis is a relatively frequent disease in Croatia. Elevated body temperature, anemia and haemoglobinuria are the most common signs. Diagnosis is rapidly obtained by employing blood smears, as B. canis is present in the red blood cells of affected dogs. Treatment is favourable and without consequences. Blood work was performed initialy, prior to treatment, and on the 1st and the 7th day following treatment. Following history and examination of the dogs blood and urine samples were taken. After confirmation of B. canis in the red blood cells, alltogether 226 dogs were tretated. Out of them 80 were tretaed with Berenil® (diminazen aceturate, Hoechst), 72 were tretated with Imizol® (imidocarb dipropionate, Schering-Plough-Animal-Health) and 74 with Oxopirvedin® (fenamidine dizetionate, Merial). Clinical findings, haematological analysis and urine analysis are given and statistically assesed. After tretment with Berenil®, symptoms of babesiosis regressed within 24 hours. Health improved more slowly in the group treated with Oxopirvedin® in comparioson with the group treated with Berenil®. Contrary to the above, Imizol® displayed the slowest regression of the disease and reinfestation with B. canis within 30 days was not noted. That is not the case if treatment was provided by Berenil® and Oxopirvedin®. In all 226 cases of canine babesiosis side effects were not noted, except topically inflammed tissue at the site of subcutaneous application.

Extracellular matrix supports healing of transected rabbit Achilles tendon

Extracellular matrix (ECM) has been intensively used in cardio surgery. The main goal of this research was to determine if Achilles tendon healing could be promoted by applying extracellular matrix scaffold (CorMatrix®, USA). Sixteen (n = 16) New Zealand white mature rabbits (Oryctolagus cuniculus) were randomly allocated into two groups. Following complete surgical transection, rabbits in group A (ECM applied) (n = 8) had their Achilles tendons reconstructed using both, nylon suture and extracellular matrix scaffold, whereas in group B (without ECM) the tendons were reconstructed using nylon suture only. After four weeks, the rabbits were euthanized and tendon samples harvested and stained with hematoxylin eosin, Mallory, and Gomory and subsequently histologically analyzed according to modified Bonnar scale. Group B had significantly stronger inflammatory response, including abundant cell infiltration and neovascularization. In group A collagen fibers were predominantly found, whereas in group B reticular fibers were more abundant. Extracellular matrix scaffold has been found to have the real potential for promoting tendon healing through accelerating collagen formation, which is crucial for restoring biomechanical properties of a tendon, decreasing peritendineous adhesion formation, and reducing inflammatory edema and subsequently pain.

Osteogrow: A Novel Bone Graft Substitute for Orthopedic Reconstruction

Complications associated with the clinical use of BMP2 and BMP7 result from the limited understanding of their molecular mechanisms in bone remodeling. Recently, a novel BMP6-based approach has been developed with superior healing results and reduced side effects in preclinical studies. BMP6-containing osteogenic medicinal product called Osteogrow, which is aimed to induce and accelerate bone formation, is currently being tested in clinical studies. It comprises of a biologically compatible autologous carrier made from the patient’s peripheral blood and of rhBMP6 as an active ingredient. Such formulation circumvents the use of animal-derived materials, significantly limits inflammatory processes common in commercial bone devices, and renders the carrier flexible and injectable ensuring the ease of use. The ongoing clinical trial results will provide a more detailed insight into the safety, tolerability, pharmacokinetics, and bone healing effects in humans and hopefully provide novel and valuable therapeutic options in the field of bone regeneration.

Osteosynthesis of the radius and ulna in a brown hare lepus europeus

Six months old brown hare was presented at the Surgery Clinic for evaluation of a right forelimb third grade lameness. Full physical examination, blood samples for complete blood count, and serum biochemistry were performed. Radiographic examination from anteroposterior and lateral views confirmed a old, closed, complete, non-comminuted, transverse fracture in the middle part of right radius and ulna. Fracture repair in the brown hare is a similar to those in cats and dogs. Stabilization of fractures with bone plates and screws are the most popular methods of fracture fixation. Osteosynthesis was performed by 2.0, 8-hole stainless steel Synthes dynamic compression plate, DCP (Synthes Holding AG, 4500 Solothurn, Switzerland) placed on radius in neutral position to achieve anatomical repositioning of fragments and to restore normal biological function of the limb. Surgical procedure was performed within 45 minutes in order to reduce residence in a hospital environment, which hares generally find stressful. The fracture was assessed as clinically stable in the 5th day postoperatively and the animal was weightbearing on its forelimb. Osteosynthesis of hare radius using DCP plate 2.0 is a good method of fixation and healing fractures. After performing full clinical examination and bloodwork, controled and modulated anaesthesia and analgesia, it is possible to perform osteosynthesis with excellent outcome and also a good way to preserve fragile wildlife.

The efficacy of milk ejection induced by luteal oxytocin as a method of early pregnancy diagnostics in cows

The aim of this research was to determine the accuracy and reliability of early diagnosis of pregnancy in cows using PGF2α analogue. Namely in the case of corpus luteum (CL) presence, the release of luteal oxytocin is induced by intravenous administration of a non luteolytic dose of PGF2α. The research was performed in 30 lactating cows, 18-22 days after insemination, and three hours prior to evening milking. A teat cannula is placed in the left fore-teat. When the cisternal milk flow ceased, Dinoprost (256 μg Dinolytic) was injected in the v. jugularis. If CL was present, the alveolar milk flow (559.4±83.7 mL) started 189±18 s later and early pregnancy could be presumed. If CL was no longer functional, the milk flow did not start again and the cow was considered not pregnant. Seventeen out of 30 cows were confirmed pregnant 35 days after insemination by ultrasound and 13 were found not pregnant. The accuracy of positive results is 100% and is identical to that obtained by ELISA of milk progesterone used as a reference method. The accuracy of negative results was lower (46.1% vs. 84.6%) (p<0.05), because seven out of 13 non pregnant cows has responded to the milk ejection test. In spite of this disadvantage, the test approved as being inexpensive, rapid, and easy to interpret and is therefore very applicable, especially under field conditions. It can be applied i.e. 20 days after insemination. Progesterone rapid test from milk or by milk ejection test can be used with similar accuracy.