A streak disease of pearl millet caused by a leafhopper-transmitted geminivirus

The cause of a streak disease of pearl millet (Pennisetum glaucum), originating from Nigeria, has been attributed to a geminivirus belonging to the African streak virus cluster. A full-length, infectious clone of the virus was obtained which was transmissible by the vectorCicadulina mbila (Naudé). Analysis of the complete nucleotide sequence of the coat protein gene of this virus shows it to be most closely related to sugarcane streak virus. The possible evolutionary implications of this finding are discussed.     

Characterisation of a maize-infecting potyvirus from Spain

In order to characterise and classify an unknown maize-infecting potyvirus isolated from fields in northeast Spain, the entire coat protein gene and the C-terminal twothirds of the large nuclear inclusion protein (NIb) gene were cloned and sequenced. Protein sequencing enabled the cleavage site between the two proteins to be deduced and also revealed that on storage the viral coat protein undergoes a specific degradation in which the N-terminal 39 amino acids are removed. Comparison of the nucleotide sequence of the 3 non-coding region of the viral RNA and the predicted amino acid sequence of the coat protein with the equivalent regions of other members of the potyvirus group revealed that the Spanish virus is closely related to maize dwarf mosaic virus strain A.     

Chylothorax associated with cryptococcal mediastinal granuloma in a cat

A 10-year-old neutered female cat had chylothorax, precaval syndrome, and a mediastinal granuloma resulting from infection with Cryptococcus neoformans. Diagnosis of a chylous effusion was made by cytologic examination of pleural fluid and by finding higher triglyceride levels in the effusion than in serum (825 vs. 64 mg/dl, respectively). Postmortem examination revealed cryptococcal organisms in the mediastinal granuloma, lungs, cerebral meninges, and connective tissues adjacent to the thyroid gland. Chylous effusion in a cat associated with cryptococcosis has not been reported previously. Cryptococcosis should be included in the differential diagnosis in chylous effusions in cats.     

‘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.     

Sexual compatibility types of Bremia lactucae isolates originating from Lactuca serriola

Results are given on the occurrence of sexual compatibility types of seven isolates ofBremia lactucae originating fromLactuca serriola (prickly lettuce). It is concluded that the isolates studied are heterothallic. Both compatibility types (B1 en B2) were determined, but type B2 was prevalent. Sexual recombination ofB. lactucae isolates originating from wild and cultivated lettuce may occur.Samenvatting Zeven isolaten vanBremia lactucae, afkomstig vanLactuca serriola in Tsjechoslowakije, zijn onderzocht op hun sexuele compatibiliteitstype door ze te combineren met Nederlandse fysio's vanB. lactucae, afkomstig van cultuursla (L. sativa), waarvan het compatibiliteitstype (B1 of B2) bekend is. Alle isolaten vanL. serriola bleken heterothallisch te zijn, waarbij type B2 meer werd aangetroffen dan type B1. Sexuele recombinatie vanBremia-isolaten van wildeLactuca-soorten en cultuursla blijkt goed mogelijk te zijn.     

How to perform transvenous electrical cardioversion in horses with atrial fibrillation

Electrical cardioversion of atrial fibrillation is a well-established technique for restoration of sinus rhythm in humans. While transthoracic cardioversion is more commonly used, transvenous electrical cardioversion (TVEC) has been reported as having higher efficacy at substantially lower energy levels. In horses, treatment of atrial fibrillation has essentially been limited to the administration of quinidine salts either orally or intravenously. TVEC provides an alternative to quinidine salts, especially for those animals in which quinidine is neither effective nor tolerated. The present report details this technique in horses, discusses possible complications of the procedure, and provides guidance for successful outcome. Still and video images are used to illustrate details with regard to TVEC techniques in horses. Please view supplemental material for the videos.     

Effects of phosphodiesterase inhibitor-induced insulin resistance on glucose and lipid metabolism in rats

AIM:To investigate the effects of milrinone (a selective phosphdiesterase III inhibitor PDE₃) on insulin secretion, blood glucose, plasma free fatty acids (FFA) and dose-response relationship, and assess possible effects of milrinone on glucose metabolism and insulin sensitivity in conscious rats.METHODS:The catheterized nonstressed rats were administered by the varying doses of milrinone (1, 5, 25 μmol/kg) and were compared with controls. A hyperinsulinaemic- euglycaemic clamp was established in awake rats, and milrinone(25 μmol/kg) and 25% dimethyl sulfoxide (DMSO, as a control) were given at 120 min during hyperinsulinaemic- euglycaemic clamp. Glucose turnover was decided by gas chromatograph mass spectrometer (GC-MS).RESULTS:After dosing, plasma FFA levels in 3 milrinone groups significantly increased compared with the controls and before dosing. The percentages of elevation of FFA by the different milrinone doses were very similar, 50%, 52%, 55% for 1, 5, 25 μmol/kg respectively at 2 min after dosing. Plasma insulin levels were significantly elevated in the 5 and 25 μmol/kg groups, and the effect of milrinone on glucose concentration was detectable only 25 μmol/kg group. During hyperinsulinaemic clamp, there were significant increase in plasma FFA (from 173.1±15.2 to 633.8±87.3 μEq/L) and hepatic glucose production (HGP), and a significant decrease in glucose infusion rates (GIR) (to about 21%).CONCLUSION:These data suggest that milrinone impaires the abilities of insulin to suppress lipolysis and HGP, and insulin-mediated glucose utilization in peripheral tissue. Therefore, milrinone administration may induce an acute insulin resistancein vivo.