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.     

Effect of progesterone on ovulation in the hypophysectomised hen

1. One hundred and seven laying hens were hypophysec‐tomised to clarify the relationship between hypophysectomy and the effect of progesterone (P4) on ovulation.

2. Hens were hypophysectomised at 6 intervals from 4.5 to 11.0 h before the expected time of ovulation. Ovulation occurred in the hens operated on from 4.5 to 7.3 h, but did not take place in birds operated on from 9.2 to 9.8 h before the expected time of ovulation.

3. When a single dose of P4 (2 mg/hen) was injected iv immediately after the removal of the anterior pituitary from 7.5 to 9.8 h before the expected time of ovulation, ovulation was induced. However, the percentage of hens responding decreased in proportion to the lapse of time between the hypophysectomy and the expected time of ovulation. No ovulation was induced in hens which were hypophysectomized and given P4 10.2 to 11.0 h before the expected time of ovulation.

4. It is suggested that ovulation is induced by P4 alone possibly in the absence of preovulatory gonadotrophins and that P4 acts directly on the ovary to induce follicle rupture.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002