Plasma digoxin concentration in dogs with mitral regurgitation.

Fifteen and eight mature beagles, without (normal group) and with experimental mitral regurgitation (MR group), respectively, were given 0.02 mg/kg/day digoxin powder for 10 days orally. The optimum time for sample collection after administration of digoxin was observed to be 8-18 hr and 10-22 hr in the normal and MR groups, respectively. In both groups, a stable concentration was reached after 3-5 days of treatment. No differences in plasma level were observed between sexes. The optimum concentration of digoxin was attained at an earlier stage than has been previously reported for both dogs and humans.     

Influence of Body Weight and Food Density on Kinetics of Ammonia Excretion by the Brine Shrimp Artemia franciscana

This study was undertaken to quantify the relationships between the rate of ammonia excretion by brine shrimp and dry weight (body size) and food density. Measurements of the rate of ammonia excretion of the brine shrimp were made at 25 C in darkness. Under these conditions, the excretion rate of ammonium-nitrogen can be generalized as a function of body weight and food (Nannochloropsis sp.) density. The relationship between the body weight (W) and excretion rate of ammonium nitrogen (E_N) is expressed as E_N=α W^γ, where α is the metabolic level. The parameter α is dependent on ration, such that α=α_o±α_r where α_o= metabolic level in starvation and α_r= the metabolic level which increased with feeding or changed with the food density. In experiments in which the animal starved, the equation above becomes E_N=α_oW^γ, and the values of α and γ were 0.22 and 0.93, respectively. The rate of ammonium excretion rose from 5.42 × 10^?4 to 2.28 × 10^?1μg N/animal/h as dry weight measured from 1.57 × 10^?3? 1.04 mg dry weight per animal. Next, it is convenient to express α_r as α_r=α_fmax= (1 ? 10^?kc), where α_fmax is the maximum rate of α_f at saturated level of food density, C is the mean cell density, and k is a constant defining the rate of change of the metabolic rate with cell density. Therefore, the rate of ammonia excretion by brine shrimp could be expressed as E_N=α_o [1 ? a(l ? 10^?kC)]W^γ, and a =α_fmax/α_o. The values of the kinetics parameters obtained from experiments where shrimp were fed were a = 2.3 and k = 0.15×10^?7. Consequently, the maximum rate of ammonia excretion at a saturated food density is equivalent to 3.3 times the rate of animals that are not fed.     

Identification of c-kit mutations-independent neoplastic cell proliferation of canine mast cells

Gain-of-function mutations in the proto-oncogene c-kit have been considered the molecular mechanism of neoplastic proliferation of mast cells. However, the importance of c-kit gene mutations is not well evaluated in canine mast cell tumors (MCTs). In the present study, we established and characterized a mast cell line, HRMC, derived from a dog with MCT. We also examined c-kit mutations in HRMC cells and assessed an inhibitory effect of a tyrosine kinase inhibitor, STI571, on HRMC cells. HRMC cells had cytoplasmic metachromatic granules, chymase and tryptase, and expressed both KIT and FcepsilonRI on the cell surface. HRMC cells contained histamine and released beta-hexosaminidase through FcepsilonRI cross-linking and calcium ionophore stimulation. Nucleotide sequence analysis demonstrated no mutations in an open reading frame of c-kit cDNA and genomic DNA of the juxtamembrane domain of c-kit in HRMC cells. STI571 did not show any inhibitory effects on the proliferation of HRMC cells. These findings clearly demonstrated the existence of c-kit mutations-independent neoplastic canine mast cell proliferation. The growth factor-independent mast cell line established in this study might be valuable to explore novel mechanisms of c-kit mutations-independent neoplastic proliferation of mast cells in dogs.     

Direct penetration of Rhizopus stolonifer into stone fruits causing rhizopus rot

Rhizopus rot, caused by Rhizopus stolonifer, is a major postharvest disease of stone fruits. The disease is related to the occurrence of mechanical and physical damage; however, observations at a Brazilian wholesale market suggest that direct penetration can occur. Therefore, the penetration mechanisms of R. stolonifer in stone fruits were evaluated. To identify the production of enzymes that help with direct penetration by the pathogen, esterase activity, both in mycelial discs and in spore suspensions of the fungus in water and in modified Van Etten nutrient solution, was measured. Assays were also conducted to evaluate the growth of R. stolonifer on glucose or cutin as a sole carbon source. The pathogen grew on both media, and higher esterase activity was observed in the cutin medium. Wounded and unwounded peaches and nectarines were inoculated with R. stolonifer spore suspensions in water or in modified Van Etten nutrient solution. Wounded fruit inoculated with either of the R. stolonifer spore suspensions developed rhizopus rot, whereas unwounded fruit developed the rot only in the presence of spores in the modified Van Etten nutrient solution. Scanning electron and light microscopic examination showed the fungus can directly penetrate the nectarine cuticle. Diisopropyl fluorophosphate, a serine hydrolase inhibitor, prevented rot development in peaches. The results provide valuable evidence for the ability of R. stolonifer to directly penetrate unwounded stone fruits, probably due to the production of esterase enzymes.     

Study of the Structure–Activity Relationship of an Anti-Dormant Mycobacterial Substance 3-(Phenethylamino)Demethyl(oxy)aaptamine to Create a Probe Molecule for Detecting Its Target Protein

The current tuberculosis treatment regimen is long and complex, and its failure leads to relapse and emergence of drug resistance. One of the major reasons underlying the extended chemotherapeutic regimen is the ability of Mycobacterium tuberculosis to attain a dormant state. Therefore, the identification of new lead compounds with chemical structures different from those of conventional anti-tuberculosis drugs is essential. The compound 3-(phenethylamino)demethyl(oxy)aaptamine (PDOA, 1), isolated from marine sponge of Aaptos sp., is known as an anti-dormant mycobacterial substance, and has been reported to be effective against the drug resistant strains of M. tuberculosis. However, its target protein still remains unclear. This study aims to clarify the structure–activity relationship of 1 using 15 synthetic analogues, in order to prepare a probe molecule for detecting the target protein of 1. We succeeded in creating the compound 15 with a photoaffinity group that retained antimicrobial activity, which proved to be a suitable probe molecule for identifying the target protein of 1.     

Pecking behaviour of laying hens in single-tiered aviaries with and without outdoor area

1. The objective of the present study was to examine the behaviour of laying hens in single-tiered aviaries with and without outdoor areas with particular reference to the proportion of each behaviour and the ways it changed. 2. In all, 144 interbred cross layers (WL/RIR cross-breed) were used. At the age of 16 weeks, the hens were divided at random into two groups and moved to single-tiered aviary (SA) and free-range systems (FR, SA with in addition an outdoor range area covered with clover) with 18 hens per pen. Behavioural observations were conducted before, during and after access to the range. 3. All behaviours using the beak (eating, grazing, drinking, preening, aggressive pecking, feather pecking, litter pecking, object pecking and mate pecking) were recorded as pecking behaviour. 4. While most of the FR hens spent their time outside foraging, the proportion of hens eating, preening, litter pecking, object pecking, aggressive pecking and feather pecking was higher in SA than in FR hens. 5. The proportion of hens performing pecking behaviour of all types was very similar in SA (61.7 +/- 2.0%) and in FR (64.0 +/- 0.8%). The proportion of hens performing overall pecking behaviour increased as pre-laying sitting decreased. 6. The proportion of hens feather pecking decreased in FR during access to range and a similar tendency was found for aggressive pecking. 7. In conclusion, the total proportion of hens pecking was almost the same regardless of whether an outdoor area was provided or not, but the incidence of different types of pecking behaviour differed between SA and FR. The risk of feather pecking in FR may be lower when an outdoor grazing area is provided, although further testing on a larger scale would be essential.     

Molecular cloning, chromosomal location, and biological activity of porcine interleukin-21

A pig interleukin-21 (IL-21) cDNA was successfully cloned and sequenced from porcine peripheral blood lymphocytes (PBL) stimulated with 10 microg/ml concanavalin A (ConA), 10 microg/ml phytohemagglutinin P (PHA), 50 ng/ml phorbol 12-myristate 13-acetate (PMA), and 0.5 microg/ml anti-porcine CD3 antibody for 48 hr. The open reading frame of the porcine IL-21 cDNA is 459 base pairs in length and encodes 152 amino acids. The predicted amino acid sequence of the porcine IL-21 shows 86.2%, 77.7%, and 58.4% identity to the bovine, human, and murine IL-21, respectively. The porcine IL-21 gene was mapped to porcine chromosome 8 (8q22-->q23) by means of fluorescence in situ hybridization and radiation hybrid mapping, where the porcine IL-2 gene had been mapped nearby. The recombinant porcine mature IL-21 expressed by E. coli induced dose-dependent proliferation and IFN-gamma production from a human NK cell line, NK0. The porcine IL-21 identified in this study will be helpful for the enhancement of innate immune responses of pigs.     

Comparison of the activities of enzymes related to glycolysis and gluconeogenesis in the liver of dogs and cats.

Activities of enzymes related to glucose metabolism were measured in canine and feline liver. There were no significant differences in plasma glucose and immunoreactive insulin concentrations between dogs and cats. Glucokinase activities were absent in feline liver, however, activities of other glycolytic enzymes such as hexokinase, phosphofructokinase and pyruvate kinase, were significantly higher than those in canine livers. Activities of rate limiting enzymes of gluconeogenesis such as pyruvate carboxylase, fructose-1, 6-bisphosphatase and glucose-6-phosphatase in feline livers were significantly higher than those in canine livers.     

Appearance of slow-reacting and complement-requiring neutralizing antibody in cattle infected with Akabane virus

Slow-reacting complement-requiring neutralizing (NT) antibody was detected in sera from cattle 2 weeks after infection with Akabane virus. Bovine sera obtained 3 or 4 weeks after infection contained slow-reacting noncomplement-requiring NT antibody. The slow-reacting complement-requiring NT antibody was sensitive to 2-mercaptoethanol (2-ME), whereas the slow-reacting noncomplement-requiring NT antibody was resistant to 2-ME. The initial phase may represent the IgM response and the later phase a change to IgG. A NT test was developed in which virus-serum mixtures were incubated at 4 degrees C for 48 h and then with complement at 37 degrees C for 60 min; this gave an improved sensitivity over the previous incubation at 37 degrees C for 60 min.