The [Ru(Hedta)NO](0.1-) system: structure, chemical reactivity and biological assays

The [Ru(II)(Hedta)NO(+)] complex is a diamagnetic species crystallizing in a distorted octahedral geometry, with the Ru-N(O) length 1.756(4) A and the RuNO angle 172.3(4) degrees . The complex contains one protonated carboxylate (pK(a)=2.7+/-0.1). The [Ru(II)(Hedta)NO(+)] complex undergoes a nitrosyl-centered one-electron reduction (chemical or electrochemical), with E(NO+/NO)=-0.31 V vs SCE (I=0.2 M, pH 1), yielding [Ru(II)(Hedta)NO](-), which aquates slowly: k(-NO)=2.1+/-0.4x10(-3) s(-1) (pH 1.0, I=0.2 M, CF(3)COOH/NaCF(3)COO, 25 degrees C). At pHs>12, the predominant species, [Ru(II)(edta)NO](-), reacts according to [Ru(II)(edta)NO](-)+2OH(-)-->[Ru(II)(edta)NO(2)](3-), with K(eq)=1.0+/-0.4 x 10(3) M(-2) (I=1.0 M, NaCl; T=25.0+/-0.1 degrees C). The rate-law is first order in each of the reactants for most reaction conditions, with k(OH(-))=4.35+/-0.02 M(-1)s(-1) (25.0 degrees C), assignable mechanistically to the elementary step comprising the attack of one OH(-) on [Ru(II)(edta)NO](-), with subsequent fast deprotonation of the [Ru(II)(edta)NO(2)H](2-) intermediate. The activation parameters were DeltaH(#)=60+/-1 kJ/mol, DeltaS(#)=-31+/-3 J/Kmol, consistent with a nucleophilic addition process between likely charged ions. In the toxicity up-and-down tests performed with Swiss mice, no death was observed in all the doses administered (3-9.08 x 10(-5) mol/kg). The biodistribution tests performed with Wistar male rats showed metal in the liver, kidney, urine and plasma. Eight hours after the injection no metal was detected in the samples. The vasodilator effect of [Ru(II)(edta)NO](-) was studied in aortic rings without endothelium, and was compared with sodium nitroprusside (SNP). The times of maximal effects of [Ru(II)(edta)NO](-) and SNP were 2 h and 12 min, respectively, suggesting that [Ru(II)(edta)NO](-) releases NO slowly to the medium in comparison with SNP.     

Expression and Stability of Arabidopsis CDC6 Are Associated with Endoreplication

Studies on the CDC6 protein, which is crucial to the control of DNA replication in yeast and animal cells, are lacking in plants. We have isolated an Arabidopsis cDNA encoding the AtCDC6 protein and studied its possible connection to the occurrence of developmentally regulated endoreplication cycles. The AtCDC6 gene is expressed maximally in early S-phase, and its promoter contains an E2F consensus site that mediates the binding of a plant E2F/DP complex. Transgenic plants carrying an AtCDC6 promoter-beta-glucuronidase fusion revealed that it is active in proliferating cells and, interestingly, in endoreplicating cells. In particular, the extra endoreplication cycle that occurs in dark-grown hypocotyl cells is associated with upregulation of the AtCDC6 gene. This was corroborated using ctr1 Arabidopsis mutants altered in their endoreplication pattern. The ectopic expression of AtCDC6 in transgenic plants induced endoreplication and produced a change in the somatic ploidy level. AtCDC6 was degraded in a ubiquitin- and proteosome-dependent manner by extracts from proliferating cells, but it was degraded poorly by extracts from dark-grown hypocotyl endoreplicating cells. Our results indicate that endoreplication is associated with expression of the AtCDC6 gene and, most likely, the stability of its product; it also apparently requires activation of the retinoblastoma/E2F/DP pathway. These conclusions may apply to endoreplicating cells in other tissues of the plant and to endoreplicating cells in other eukaryotes.     

Preferences for Call Spectral Properties in Hyla intermedia

We studied the pattern of variation in the spectral properties of male advertisement calls and female preference functions for these same properties in the Italian treefrog (Hyla intermedia). The call spectral properties (fundamental and dominant frequencies) were highly negatively correlated with body size, they showed low within‐individual variation, and, at the population level, they were found to be under weak stabilizing selection. In two‐choice discrimination tests, females did not show preferences between calls with fundamental and dominant frequencies within two standard deviations from the population mean, whereas females significantly discriminated against calls when their frequencies were three (or more) standard deviations above or below the population mean. Consistent with the observed permissiveness in female preference over the call spectral structure, in the natural population, we found no evidence either of directional selection on male body size or of a size‐assortative mating pattern. The pattern of preferences observed at the population level did not mirror the preferences of individual females. In fact, in a multi‐trial discrimination experiment, females showed significant differences in their choice between the average frequency call and a two standard deviation lower than average frequency alternative, for which no significant preferences were observed at the population level. Variations in preference were not found to correlate with female body size.     

Agrobacterium rhizogenes—transformed plant roots as a source of grapevine viruses for purification

Agrobacterium rhizogenes-mediated transformation was applied toVitis spp. andNicotiana spp. infected by different grapevine phloem-limited viruses (grapevine fleck virus, grapevine virus A, grapevine virus B) to obtain root cultures for virus purification. All plant species were successfully transformed, and several clones were established in liquid culture. Transformed grapevine roots contained as much virus as non transformed roots and more than leaves, as assessed by ELISA and thin sectioning. Likewise, transformed roots ofNicotiana benthamiana Domin. contained in average more GVA than leaves, especially those at the base and the top of the plant, whereas withNicotiana occidentalis wheel., GVB was apparently less concentrated than in leaves.Nicotiana root grew faster than those ofVitis. All viruses multiplied and persisted in root cultures, which were successfully used for purification. Virus yields were the same (GFkV and GVB) or higher (GVB) than those reported in the literature. Grapevine roots may prove useful for culturing and purifying other non-mechanically transmissible grapevine viruses.     

Monitoring of airborne pollen and pollen calendar of Cosenza,Southern Italy

Summary From May 1986 to May 1989 surveys of airborne biological particles have been performed in the atmosphere of Cosenza, Italy with the aim of monitoring the presence of airborne pollen. The survey station is situated at Arcavacata of Rende, a hilly area 474 meters above sea level, 7 km north-west of the main town. The sampler (VPPS 2000) is located about 20 meters above ground level. The monitoring, performed in accordance with the criteria suggested by the Italian Association of Aerobiology (A.I.A.), enabled the identification of 26 different pollen types. The pollination graphics show: 1) a late winter period in which pollen from trees appears; 2) a spring period with a prevalence of Gramineae, Fagaceae, Oleaceae andParietaria pollen; 3) a late spring-summer period during which Fagaceae,Pinus and Compositae pollen grains are present; 4) a late summer-autumn and winter period characterized by a drastic reduction of airborne pollen. A study of the calendar reveals, furthermore, that: 1) Gramineae pollen is, amongst those of allergological importance, the most representative; 2)Parietaria is not perennial and is present in modest concentrations; 3)Olea reaches a high peak in June, but not as high as in other southern regions; 3) pollen from arboreal plants is prevalent compared to that of herbaceous plants.     

Effect of exogenous FSH on estrus, ovulation and endogenous hormone release in dairy cows

Fifteen lactating Holstein cows were randomly allotted to receive either 0 mg (group 0), 32 mg (group 1) or 50 mg (group 2) porcine follicle stimulating hormone (FSH-P) injected in 10 fractions at 12 hr intervals beginning on day 9 of the estrous cycle. All cows received 25 mg prostaglandin (PG) on day 11. Jugular blood samples were collected from cows in all groups at 6 hr intervals beginning on day 7 and continuing through expression of estrus. Mean duration to occurrence of estrus and preovulatory LH surge after PG injection was reduced (P<.05) by injection of FSH-P. Mean number of ovulations increased (P<.05) progressively with increased dose of FSH-P. Mean peripheral progesterone declined more uniformly in FSH-P treated cows after PG and increased earlier (P<.05) after estrus in group 2 cows compared to group 0 and 1 cows. Mean plasma estradiol-17beta elevated (P<.05) after PG injection in both FSH-P-treated groups compared to group 0 cows. Both LH and FSH increased (P<.05) for 36 hr after initiation of FSH-P injection in groups 1 and 2, then declined until after PG injection. Peak LH and FSH occurred more uniformly following PG in treated cows. Results indicate that FSH-P increased endogenous gonadotropin release, estradiol-17beta, ovulation rate and reduced duration to estrus and preovulatory gonadotropin release after PG. Injection of 50 mg FSH-P increased plasma estradiol-17beta and ovulation rate compared to injection of 32 mg FSH-P.