Molecular characterization of Iranian wheat stripe virus shows its taxonomic position as a distinct species in the genus Tenuivirus.

The full lengths of three genome segments of Iranian wheat stripe virus (IWSV) were amplified by reverse transcription (RT) followed by polymerase chain reaction (PCR) using a primer complementary to tenuivirus conserved terminal sequences. The segments were sequenced and found to comprise 3469, 2337, and 1831 nt, respectively. The gene organization of these segments is similar to that of other known tenuiviruses, each displaying an ambisense coding strategy. IWSV segments, however, are different from those of other viruses with respect to the number of nucleotides and deduced amino acid sequence for each ORF. Depending on the segment, the first 16-22 nt at the 5' end and the first 16 nt at the 3' end are highly conserved among IWSV and rice hoja blanca virus (RHBV), rice stripe virus (RSV) and maize stripe virus (MStV). In addition, the first 15-18 nt at the 5' end are complementary to the first 16-18 nt at the 3' end. Phylogenetic analyses showed close similarity and a common ancestor for IWSV, RHBV, and Echinochloa hoja blanca virus (EHBV). These findings confirm the position of IWSV as a distinct species in the genus Tenuivirus.

Induction of Spodoptera littoralis nuclear polyhedrosis virus in cell cultures.

A nuclear polyhedrosis virus (NPV) infection was initiated in a Spodoptera littoralis cell line when haemocytes from granulosis virus (GV)-infected S. littoralis larvae and ultra-violet-inactivated Sendai virus were added.

Reduced pyridine nucleotide oxidases of Eugena gracilis var. bacillaris.

Cell-free extracts of a streptomycin-bleached strain of Euglena gracilis var. bacillaris have been examined for enzyme systems primarily responsible for the oxidation of reduced pyridine nucelotides. NADH lipoyl dehydrogenase, NADH and NADPH oxidase, NADH and NADPH diaphorase, and NADH and NADPH cytochrome c reductase have been demonstrated. The NADPH-linked enzymes had lower activity rates and were less sensitive to N-ethyl maleimide and p-hydroxymercuribenzoate than their NADH-linked counterparts. NADH cytochrome c reductase was the most sensitive to antimycin A. Michaelis-Menten constants (Km) determined were as follows: NADH diaphorase, 350 muM; NADPH oxidase 150 muM ; NADH lipoyl dehydrogenase, 0.35 muM. Enzyme activities after storage at -5 C indicate that the diaphorases are less labile than the other tested enzymes, and the differential activities of the NADH and NADPH linked enzymes suggest that functionally they may have different roles.

Comparative study of the thiourea carrier in erythrocytes.

A densimeter technique was used to measure the rate of exit of thiourea from erythrocytes of various species of mammals. The cells were first equilibrated with a 200 mM thiourea solution in 1% NaCl. An aliquot of these cells was added to 1% NaCl containing 4.6-23.1 mM thiourea. Facilitated diffusion was demonstrated in each case. Using exit times or initial rates, calculations of half-saturation constants (phi) in mM and maximum transport rates (K) in isotones per min were made by three different methods. The following values were obtained: human-phi=60, 42, 35; K=1.2, 2.9, 0.9; rabbit-phi=46, 33, 32; K=0.8, 2.1, 0.8; mouse-phi=46, 40, 30; K=3.4, 8.5, 3.2; rat-phi=65, 42, 23; K=6.1, 15.3, 3.7; ox-phi=107, 63, 88; K=0.6, 1.4, 0.4; sheep-phi=56, 38, 56; K=0.9, 2.2, 0.6; and pig-phi=110, 64, 49; K=1.6, 3.6, 1.1.

Permeability of trout erythrocytes to nonelectrolytes.

Using a densimeter technique, a kinetic analysis was made, employing both entrance and exit studies, of the permeability of erythrocytes of brook trout (Salvelinus fontinalis), rainbow trout (Salmo gairdneri), German brown trout (Salmo trutta) and cutthroat trout (Salmo clarki) to glycerol, ethylene glycol, thiourea and urea. All of the data indicate that these four nonelectrolytes cross the membrane of the erythrocytes of these four species of fishes by simple diffusion only.

Measurements of exit rates to distinguish between facilitated and simple diffusion.

A modification of the method of Sen and Widdas (J. Physiol. London 160:392-403, 1962) was used to measure the rate of exit of several nonelectrolytes from erythrocytes of various species. In spite of additional errors introduced by the larger half-saturation values of the carriers (phi) and concentrations, it was possible to distinguish between systems with small values of phi, systems with relatively large values of phi, and systems involving only simple diffusion. Approximate values of phi in millimoles and of maximum transfer rate (K) in isotonic units per minute were obtained using times and initial slopes measured on experimental curves. The following values in the foregoing units were obtained: human glucose, phi = 1.8, 1.0, K = 0.8, 1.1; human glycerol, phi = 178, 94, K = 4.3, 3.3; sheep thiourea, phi = 56, 56, K = 0.9, 0.6; and rabbit glycerol, phi = 328, 64, K = 2.2, 1.0. Simple diffusion was demonstrated for the following systems: ox-ethylene glycol; ox-glycerol; sheep-ethylene glycol; and sheep glycerol.

Kinetic analysis of the permeability of human erythrocytes to NH4C1.

A densimeter technique was used to make a kinetic analysis of the rate of swelling of human erythrocytes suspended in 1% NaCl following successive additions of NH(4)Cl. Two series of experiments were performed, one in the absence of and the other in the presence of 6 x 10(-4)M NaHCO(3). An analysis of the data using Widdas's equations gave a family of curves in each instance. When LeFevre's equation was used with a value of ø = 1.3 isotones, a straight line was obtained with both series of data. It is concluded that this system shows carrier kinetics.