Immunohistochemical detection of P-glycoprotein (PGP) and multidrug resistance-associated protein (MRP) in canine cutaneous mast cell tumors

Fifty-four canine cutaneous mast cell tumors were evaluated immunohistochemically for the expression of P-glycoprotein (PGP) and multidrug-resistance-associated protein (MRP). All tumors examined were graded according to the histological malignancy. ranging from grade I to III. The expression of PGP was confirmed in 15% (8/54) of whole, 33% (5/15) of grade I, 10% (3/31) of grade II, and 0% (0/8) of grade III tumors. The expression of MRP was found in 18% (10/54) of whole, 26% (4/15) of grade I, 19% (6/31) of grade II, and 0% (0/8) of grade III tumors. The cases positive to at least one of these 2 multidrug markers were 26%, 47%, 23% and 0% of whole and grade I to III tumors, respectively. These results indicate that at least 26% of canine cutaneous mast cell tumors express PGP and/or MRP and that these tumors may be resistant to several anti-cancer drugs.     

Hepatic lymphangiomatosis in a young dog

A 9-month-old intact male American cocker spaniel was referred because of hepatomegaly and ascites. Ultrasonographic evaluation of the liver revealed congestion and increased parenchymal echogenicity with focal and more hyperechoic nodules. Histopathology of the hepatic lesion revealed diffuse, ill-defined vascular proliferation. A single layer of endothelial cells, which showed signs of minimal cellular atypia, lined the irregular vessels. On immunohistochemistry, the proliferative endothelial cells lining the irregular vessels were positive for an antiserum to factor VIII related antigen. Based on these findings, the dog was diagnosed with hepatic lymphangiomatosis.     

Vascular endothelial growth factor (VEGF) promotes the early development of bovine embryo in the presence of cumulus cells

Three experiments were conducted to determine the effect of Vascular Endothelial Growth Factor (VEGF) on bovine embryonic development in vitro. Human recombinant VEGF(165) was employed at 5 ng/ml in modified synthetic oviduct fluid. In Exp. 1, bovine cumulus oocyte complexes were matured with or without VEGF for 22 hr, inseminated without VEGF for 6 hr, then cultured with or without VEGF for 42 hr. The cleavage rate and the development rate to 4- to 8-cell were higher (P<0.05) in groups with VEGF during in vitro maturation (IVM, 71.4% and 59.6%), in vitro culture (IVC, 70.3% and 62.3%), and both IVM and IVC (75.9% and 67.8%) than in the group cultured thoroughly without VEGF (49.9% and 38.4%, respectively). In Exp. 2, 4- to 8-cell embryos produced in vitro without VEGF were removed from cumulus cells at 48 hr post-insemination (Pi) and cultured with or without VEGF for 144 hr. The development rates to blastocyst at 96 hr (D6), 120 hr (D7) and 144 hr (D8) were similar (P>0.05) in both groups. In Exp. 3, cumulus cells were removed from presumptive embryos produced by IVM and IVF without VEGF at 10 hr Pi. Denuded embryos were cultured with or without VEGF for 38 hr or 182 hr. The cleavage rate and the development rates to 4- to 8-cell at 48 hr Pi and to blastocyst on D6, D7 and D8 were similar (P>0.05) in all groups. These results suggest that VEGF has a beneficial effect on the initial development of bovine embryo through surrounding cumulus cells.     

Insulin responsiveness to glucose and tissue responsiveness to insulin in lactating, pregnant, and nonpregnant, nonlactating beef cows

Insulin responsiveness to glucose and tissue responsiveness to insulin, using the hyperglycemic clamp and the hyperinsulinemic euglycemic clamp techniques, were measured in lactating, late pregnant, and nonpregnant, nonlactating (NPNL) beef cows. The glucose infusion rate (GIR) in the hyperglycemic clamp technique was higher (P less than .05). in lactating cows than in NPNL cows. The plateau in plasma insulin concentration (insulin responsiveness) was higher (P less than .05) in lactating cows than in late pregnant and NPNL cows. Pregnant cows tended to have higher GIR and lower plateau in plasma insulin concentration than NPNL cows. In the hyperinsulinemic euglycemic clamp technique, GIR (tissue responsiveness to insulin) was higher (P less than .05) in lactating cows than in late pregnant cows; values for NPNL cows were intermediate. We conclude that insulin responsiveness to glucose and tissue responsiveness to insulin were enhanced during lactation but tended to be decreased during late pregnancy in beef cows.     

Effects of bacterial strains on the development of the ripening flavor of cured pork loins

To determine the effect of bacteria on the development of the ripening flavor of cured meat, pork loins were cured with pickles containing 8 or 16% sodium chloride at 0 and 8°C. The bacterial flora, total free amino acid content and total free fatty acid content in the cured loins, sensory properties of cooked pork loins and the relationships between the proteolytic and lipolytic activities of isolates and curing conditions were investigated. Desirable bacteria, including salt‐tolerant bacteria and lactic acid bacteria, were predominant under the condition of 8% sodium chloride in the pickles and a curing temperature of 8°C. The total free amino acid content and total free fatty acid content at a curing temperature of 8°C were higher than those at 0°C. Cooked pork loins cured in pickles containing 8% sodium chloride at a temperature of 8°C for more than 7 days were preferred in terms of color, flavor and taste. Before the curing procedure, gram‐negative bacteria (Vibrio, Acinetobacter, Pseudomonas and Enterobacteriaceae) were predominant in the pork loins. During the curing period, the numbers of viable gram‐positive bacteria (Micrococcus, Staphylococcus and Pediococcus) increased and the numbers of viable gram‐negative bacteria decreased. Strains of Micrococcus and Staphylococcus with proteolytic and/or lipolytic activities in cured meat also increased during the curing period and were more predominant in the pork loins cured in pickles containing 8% sodium chloride and at a curing temperature of 8°C than in pork loins cured in pickles containing 16% sodium chloride and at a curing temperature of 0°C. The actions of these bacteria were thought to be important factors affecting the flavor of the cured pork. Micrococcus and Staphylococcus with proteolytic activity might contribute to the development of the ripening flavor of ham. The results of the present study together with the results of further investigations on the relationships of the enzyme activities of Micrococcus and Staphylococcus in cured meat with a preferable flavor would be useful for establishing a novel effective method for using bacteria to produce ham of a high quality.     

Molecular biology and biotechnology for reduction of Fusarium mycotoxin contamination

Fusarium head blight (FHB) is a devastating disease of important cereal crops resulting in significant yield loss and mycotoxin contamination. Persistent outbreaks of FHB in Europe and North America have led to various efforts to understand the mechanisms of resistance to this disease and mycotoxin biosynthesis. In this minireview, we summarize basic and applied studies conducted in our laboratories into reducing mycotoxin contamination in FHB.     

Light-chain multiple myeloma in a cat.

A diagnosis of light-chain multiple myeloma was made in an 11-year-old male American Shorthair cat. The cat showed atypical plasma cell infiltration in the bone marrow, biclonal gammopathy caused by polymerization of myeloma protein (M-protein), and Bence-Jones proteinuria. The M-protein in the serum of the cat was analyzed by using 12% sodium dodeyl sulfate (SDS) polyacrylamide gel electrophoresis with Coomassie brilliant blue staining. An intense band with a size of 27 kDa, the size of the immunoglobulin light chain, was clearly observed, whereas the band corresponding to the immunoglobulin heavy chain (59 kDa) was undetectable. The 27-kDa band was confirmed to be an immunoglobulin light chain by Western blotting by using antibodies for feline immunoglobulin. These data suggested that the neoplastic plasma cells produce light chain only, leading to the diagnosis of light-chain multiple myeloma in the cat.     

Inhibitory effects of isoleucine and antagonism of the other branched-chain amino acids on fermentation parameters by mixed ruminal microbes in batch cultures and rumen simulating fermenters (Rusitec)

The inhibitory effect of isoleucine (Ile) and antagonistic effect of leucine (Leu) and valine (Val) against the inhibition by Ile were investigated using simple batch and semicontinuous cultures of mixed ruminal microbes under feeding conditions using ordinary feed (timothy hay and corn grain). In the batch culture experiment, supplementation with Ile inhibited digestion and fermentation at 8–24 h of incubation as the concentration of Ile increased up to 5 mmol/L, but these inhibitions diminished at 48 h. The inhibition caused by Ile was mitigated by the addition of Leu and Val. In a semi‐continuous culture experiment using Rusitec, digestion and fermentation of the diet was suppressed by the administration of 1 and 5 mmol/L of Ile. The daily yield of microbial‐N was also reduced by Ile, but the density of the protozoa was not. The presence of Leu and Val alleviated the inhibition by Ile. These results suggest that the dietary inclusion of Ile possibly represses the bacterial growth, feed digestion and fermentation in the rumen in ordinary feeding situations by feedback inhibition, which could be diminished in the presence of other branched‐chain amino acids. The influences of the passage and level of supplementation on inhibition by amino acids are discussed.