Expression of the Vascular Endothelial Growth Factor (VEGF‐A) and its Receptors in the Umbilical Cord in the Course of Pregnancy in the Pig

The umbilical cord (UC) and the placenta are important organs through which respiratory gases, nutrients, wastes and biologically active substances are exchanged between the maternal and the foetal system. A rapid placental vascularization observed in the second half of pig pregnancy is positively correlated with the mRNA expression of the vascular endothelial growth factor (VEGF). Based on these findings, we hypothesized that VEGF may have a stimulatory effect in the dynamically growing UC. To further understand the role of the VEGF–VEGFR system during UC development, mRNA and protein expression as well as the cellular localization of VEGF‐A, VEGFR‐1 and VEGFR‐2 in UC were examined on days 40, 60, 75 and 90 of pregnancy and after physiological delivery in the pig (day 114 of pregnancy). Real Time RT‐PCR analysis showed an increase in the mRNA levels of VEGF120 and VEGF164 from day 90 of pregnancy. VEGFR‐1 mRNA expression was significantly increased on day 75 of pregnancy. No significant changes in VEGFR‐2 mRNA expression were detected. In turn, western blot analysis revealed an increase in VEGF‐A protein expression on day 40, compared to the later days of pregnancy. A rapid increase in the VEGFR‐1 protein level was noted on day 75 and 90 of gestation. No significant changes in VEGFR‐2 protein expression were detected on any of the analysed days of pregnancy. Immunohistochemical staining enabled detection of VEGF–VEGFR system, in endothelial and tunica media cells of the umbilical vessels and in allantoic duct and amniotic epithelium on all analysed days of pregnancy. Positive reactions for VEGF‐A and VEGFR‐1, but not VEGFR‐2, were also observed in myofibroblasts. In conclusion, this data shows that members of the VEGF–VEGFR system are temporally and spatially well localized for playing key roles during umbilical cord formation and its intensive growth observed after day 75 of pregnancy.     

Serum neopterin levels in female dogs with malignant mammary tumours

In this study, we have determined serum neopterin levels in female dogs with primary malignant mammary tumours. The study involved 50 female dogs which had a malignant mammary tumours removed surgically (32 animals with carcinoma, 12 animals with sarcoma and 6 animals with carcinosarcoma) and 10 clinically healthy female dogs. Serum neopterin levels were determined using a commercial ELISA kit. The mean neopterin levels were lower in the malignant tumour groups than in healthy animals but differences were statistically significant only in carcinoma and sarcoma groups. The decrease of neopterin levels in animals with malignant mammary tumours may suggest their decreased cellular immunity. Moreover, it might indicate that decreased activity of cellular mechanisms of the anti‐neoplastic response is one of the factors associated with the development and course of malignant mammary tumours in female dogs; however, further studies are necessary.     

Comparative Evaluation of 2 In-Clinic Assays for Vector-Borne Disease Testing in Dogs

Vector-borne agents comprise medically important infections affecting dogs throughout much of the world. Sensitive detection of antibodies directed at tick-borne disease-causing organisms in dogs is diagnostically important for veterinarians, pets and their owners, and epidemiologically important for public health surveillance. The SNAP 4Dx Plus Test (IDEXX Laboratories, Inc., Westbrook, ME) identifies antibodies to or infection with multiple tick-borne pathogens and canine heartworm antigen in a single assay. Recently, VetScan FLEX4 Rapid Test (Abaxis, Inc., Union City, CA) was launched as a new assay to detect tick-borne pathogen antibodies and heartworm antigen. In the present study, we evaluated the comparative performance of SNAP 4Dx Plus (SNAP) and FLEX4 Rapid Test (FLEX4) using samples selected based on geographic distributions for canine vector borne diseases, including Borrelia burgdorferi (n = 105), Anaplasma phagocytophilum (160), Anaplasma platys (115), Ehrlichia canis (154), Ehrlichia ewingii (163), Ehrlichia chaffeensis (151) and Dirofilaria immitis (105). Canine vector borne diseases infection status was established for each sample by a combination of reference methods that included necropsy (D. immitis, heartworm disease), Western immunoblotting (B. burgdorferi), immunofluorescence assays (A. phagocytophilum and E. canis) and species-specific ELISAs (A. platys, E. canis, E. ewingii and E. chaffeensis). For comparisons among the 2 assays, samples were evaluated per the manufacturers’ instructions for each test kit.By testing each same sample set compared to the defined reference results, sensitivities differed substantially between SNAP and FLEX4, at 95.5 vs. 40.9%, respectively for B. burgdorferi, 97.1% vs. 61.4% for E. canis, 98.2% vs. 59.3% for E. ewingii, 64.3% vs. 35.7% for E. chaffeensis, 84.5% vs. 12.7% for A. phagocytophilum, 83.3% vs. 33.3% for A. platys, and 94.1% vs. 88.2% for D. immitis. Specificities for both rapid assay tests ranged from 98% to 100%.Based upon the comparative results derived from this study, the SNAP test was more sensitive than the FLEX4 test for detection of antibodies to all tick-borne pathogens and heartworm disease (Dirofilaria immitis) antigen in dogs.     

Chemically induced sterility in wheat for hybrid seed production

Summary The gametocidal effects of RH-531, RH-532, and RH-2956 were studied with three rates and times of foliar application on two spring wheat (Triticum aestivum L.) varieties, Anza and Yecora 70. RH-531 and RH-532 applied at or before booting stage caused rather large reduction in plant height while RH-2956 had a small effect on height. Application of any of the three chemicals on Anza at meiotic stages reduced self-pollination (SP) fertility by 97–99%. If 90% SP fertility reduction is acceptable, the application time could vary from premeiosis to postmeiosis for Anza. Yecora 70 was less sensitive than Anza to gametocide treatments. The lowest SP fertility obtained in Yecora 70 with RH-531, RH-532, and RH-2956 was by treatment at booting (4 kg/ha), at premeiosis (2 kg/ha), and heading (8 kg/ha), and the corresponding SP fertility reductions were 74, 89, and 71%. Pollen stainability-determined by aceto-carmine, IKI, and Alexander's stains-was not affected by gametocide treatment and was not useful in evaluating sterility induced by these chemicals. For the RH-531-treated Anza and Yecora 70 and the RH-532-treated Anza, more than 94% of the seeds produced by open-pollination on treated plants were from self-or sib-pollination as determined by progeny testing. However, RH-532-treated Yecora 70 and RH-2956-treated Anza and Yecora 70 did result in more crossed seeds, with a maximum of 20% outcrossing. It was believed that spike compactness induced by the chemicals and the related poor flower opening were the major limitations for outcrossing. Artificial pollinations made to the test varieties showed that RH-531 induced both male and female sterility and was undesirable for practical use. RH-532, however, could be applied at the booting stage to avoid severe female sterility, and RH-2956 clearly was the best of the three chemicals, with the female fertility very close to those of the untreated checks of both Anza and Yecora 70. Of the three chemicals, RH-2956 gave the highest percentage of hybrid plants in the progeny of hand-or open-pollinated spikes. Its practical use for hybrid seed production, however, will depend largely on improvement of methods to increase cross pollination in wheat.     

A COMPARISON OF DIATRIZOATE* AND IOXAGLATE† FOR POSITIVE CONTRAST SHOULDER ARTHROGRAPHY IN DOGS

In six experimental dogs, arthrographic quality and synovial inflammatory response with shoulder arthrography comparing meglumine-sodium diatrizoate (Urovison) and the monoacidic dimer, meglumine-sodium ioxaglate (Hexabrix) was evaluated. In our study initial films were of equally high diagnostic quality for both contrast media, but delayed films significantly favored ioxaglate for diagnostic quality. The rise in white blood cells in synovial fluid samples collected 24 hours after the arthrographic procedure was significantly lower after the use of ioxaglate. Histologic examination performed 14 days after the intra-articular injection revealed no drug related lesions.     

Associations between use of electric cow-trainers and clinical diseases, reproductive performance and culling in Swedish dairy cattle

The aims of this follow-up study were: (a) to evaluate the role of ECT technology as a risk factor for several diseases; and (b) to determine if the effects of these diseases on cows' reproductive performance and as risk factors for culling are influenced by exposure to ECTs. Diseases considered were retained placenta, metritis, ketosis, cystic ovaries, silent heat, milk fever, clinical mastitis, and foot and leg problems. We used historical and contemporary controls (with control herds selected to match the experimental herds for size and location). Data consisted of 10264 Swedish Red and White (SRB) and 5461 Swedish Friesian (SLB) lactation records in 150 herds of which 33 used cow-trainers. Logistic regression was used to estimate the effects of parity and exposure to electric cow-trainers on the risks of diseases and the effects of diseases and exposure to electric cow-trainers on risk of culling. The least-squares procedure was used to estimate the effects of diseases on reproductive performance.

The dominant effects associated with use of electric cow-trainers were an increased risk for silent heat, clinical mastitis, ketosis and culling relative to cows in herds not using cow-trainers. Diseases had negative effects on reproductive performance and the effects were larger for cows in herds using cow-trainers. In herds using electric cow-trainers, the largest increase in the interval from first service to conception (58 days) was caused by the occurrences of silent heat, cystic ovaries and the combination of two or more diseases. Retained placenta, metritis, cystic ovaries, clinical mastitis and a combination of two or more diseases increased the risk of culling about two times relative to healthy primiparous cows with the increase being greater for cows in herds using cow-trainers. Silent heat did not increase risk of culling in control groups, but was the largest risk factor for culling in the exposed group. We concluded that exposure to electric cow-trainers increased the incidence risk of silent heat, clinical mastitis, and ketosis and changed silent heat from a neutral disease with respect to culling to a major risk factor. Finally, exposure to cow-trainers increased the general negative effect of diseases on the cows' reproductive performance and risk for culling.     

In vitro and in vivo bioactivity of recombinant canine hepatocyte growth factor

Hepatocyte growth factor (HGF) is crucial for the development and regeneration of the liver and offers a possible new therapeutic strategy for the treatment of canine liver disease. In this study, the in vitro and in vivo bioactivity of recombinant canine HGF (rcHGF) produced with a baculoviral expression system in insect cells was measured. In vitro rcHGF induced mitogenesis, motogenesis, and phosphorylated the HGF receptor c-MET and its downstream mediators PKB and ERK1/2 in two canine epithelial cell lines. After a partial hepatectomy (phx) in dogs, rcHGF increased phosphorylation of c-MET, PKB and ERK1/2. A moderate increase was seen with the cell cycle protein PCNA in rcHGF treated livers, but no HGF-induced increase in liver weight was seen 7 days after phx. Furthermore, rcHGF treated livers showed lower levels of the key mediator of apoptosis, caspase-3, at 7days after phx. It is concluded that rcHGF is a biologically active protein in vitro and in vivo and the baculoviral expression system supplies sufficient amounts of rcHGF for future clinical studies in dogs.