Transfer of Agrobacterium DNA to Plants Requires a T-DNA Border But Not the virE Locus

Agrobacterium tumefaciens induces tumors in plants by transferring and integrating oncogenes (T-DNA) into the chromosomes of host plant cells. Agrobacterium strains were used to transfer complementary DNA copies of a potato spindle tuber viroid (PSTV) to plant cells at a wound site on tomato plant stems. Subsequently, infectious viroid RNA was found in the leaves of these plants, indicating systemic PSTV infection. This process utilized the T-DNA transfer mechanisms of Agrobacterium since PSTV infection required most virulence genes (vir) as well as one of the DNA sequences that flank either side of the Agrobacterium T-DNA. However, transfer still occurred from virE mutants of Agrobacterium, strains that fail to induce tumors even though a completely functional T-DNA is present. The virE gene seems to be directly involved in the integration of foreign DNA into plant chromosomes.     

Composition, stability, and bioavailability of garlic products used in a clinical trial

In support of a new clinical trial designed to compare the effects of crushed fresh garlic and two types of garlic supplement tablets (enteric-coated dried fresh garlic and dried aged garlic extract) on serum lipids, the three garlic products have been characterized for (a) composition (14 sulfur and 2 non-sulfur compounds), (b) stability of suspected active compounds, and (c) availability of allyl thiosulfinates (mainly allicin) under both simulated gastrointestinal (tablet dissolution) conditions and in vivo. The allyl thiosulfinates of blended fresh garlic were stable for at least 2 years when stored at -80 degrees C. The dissolution release of thiosulfinates from the enteric-coated garlic tablets was found to be >95%. The bioavailability of allyl thiosulfinates from these tablets, measured as breath allyl methyl sulfide, was found to be complete and equivalent to that of crushed fresh garlic. S-Allylcysteine was stable for 12 months at ambient temperature. The stability of the suspected active compounds under the conditions of the study and the bioavailability of allyl thiosulfinates from the dried garlic supplement have validated the use of these preparations for comparison in a clinical trial.     

Bayesian estimation of cluster-level test accuracy based on different sampling schemes

We develop Bayesian models to estimate cluster-level test characteristics, sensitivity, specificity, prevalence, and predictive values, based on four different sampling schemes: a single test case and three sequential test cases. The corresponding cluster-level characteristics are calculated and compared for different sample sizes, sampling schemes, individual-level sensitivities, specificities, and cut-off values. We compared posterior estimates of individual-level and cluster-level characteristics for these four sampling schemes with simulated data. Two illustrations, one for Johne’s disease in cattle and another for Salmonella in pig herds, are used to demonstrate application of the methods.     

Degradation of selected polycyclic aromatic hydrocarbons in coastal sediments: Importance of microbes and polychaete worms

Rates of degradation of polycyclic aromatic hydrocarbons (PAH) were examined after addition of crude oil enriched with anthracene, fluoranthene, benz(a)anthracene, and benzo(a)pyrene to coastal sediments in a laboratory (20°) flowing seawater system. Three types of sediment (fine sand, medium sand, and marsh sediment) with and without the benthic polychaete worm, Capitella capitata, were used. After extraction from the sediment, PAH concentrations were measured by liquid chromatography with fluorescent detection. Microbial degradation was studied by incubating sediments with radioactive PAH and measuring subsequent production of ¹⁴C0₂. Concentrations of the four PAH decreased significantly with time in fine and medium sized sands. PAH levels also decreased in marsh sediment but trends were not significant (P < 0.05). C. capitata stimulated PAH degradation. Microbial degradation was more rapid in upper surfaces than in lower layers of the sediments.     

Kinetic and stoichiometric assessment of the antioxidant activity of flavonoids by electron spin resonance spectroscopy

There is current interest in the use of naturally occurring flavonoids as antioxidants for the preservation of foods and the prevention of diseases such as atherosclerosis and cancers. To establish the molecular characteristics required for maximum antioxidant activity, electron spin resonance (ESR) spectroscopy has been used to determine the stoichiometry and kinetics of the hydrogen-donating ability of 15 flavonoids and d-alpha-tocopherol to galvinoxyl, a resonance-stabilized, sterically protected aryloxyl radical. The second-order reaction rates, which will be governed by O-H bond dissociation energies, were myricetin > morin > quercetin > fisetin approximately catechin > kaempferol approximately luteolin > rutin > d-alpha-tocopherol > taxifolin > tamarixetin > myricetin 3',4',5'-trimethyl ether > datiscetin > galangin > hesperitin approximately apigenin. Reactivity is highly dependant on the configuration of OH groups on the flavonoid B and C rings, there being little contribution from the A ring to antioxidant effectiveness. Highest reaction rates and stoichiometries were observed with flavonols capable of being oxidized to orthoquinones or extended paraquinones. However, rates and stoichiometries did not always correlate and the data suggest that kinetic factors may be of greater importance within a biological context.     

Preparation of tetrahydroagathic acid: a serum metabolite of isocupressic acid, a cattle abortifacient in ponderosa pine

Isocupressic acid (1) was used to synthetically prepare a mixture of (8S,13R,S)-labda-15,19-dioic acid (tetrahydroagathic acid) (5) via a two-step oxidation procedure followed by hydrogenation of the double bonds at C13 and C8. Reduction of the C8,17 double bond was stereospecific producing only the 8S isomer and confirmed by the nOe interaction between the resulting C17 and C20 methyl groups. The 13R and 13S isomers of 5 were separated and analyzed by HPLC/MS, and (13S)-tetrahydroagathic acid was isolated and identified by comparison to a standard prepared by hydrogenation of naturally occurring (13S)-dihydroagathic acid (4). (13R,S)-dihydroagathic acid was prepared by selective sodium metal-catalyzed hydrogenation of the C13,14 allylic double bond of agathic acid (3). The prepared compounds were then used as standards to confirm the presence of 4 and 5 and their respective 13R and 13S isomers in bovine serum samples. Tetrahydroagathic acid was shown to be the only metabolite detected in serum samples taken from a suspected cattle abortion case submitted for diagnosis; and, thus, 5 could be a valuable diagnostic marker for pine needle-induced abortions.     

Toxigenic type D Pasteurella multocida in New South Wales pig herds—prevalence and factors associated with infection

Between March and July 1987, a study was undertaken to determine the prevalence of and factors associated with toxigenic type D Pasteurella multocida infection in New South Wales pig herds. Toxigenic type D P. multocida was isolated from the nasal cavities of pigs in one (2%) of 50 randomly selected herds. Toxigenic isolates were also recovered from 2 (8%) of a separate group of 25 herds that had purchased pigs from a known infected piggery in South Australia (herd SA). Snout abnormalities were present in 9.4%, 3.2% and 1.8% of grower pigs in the 3 affected herds. Isolation of toxigenic P. multocida was significantly associated (p less than 0.0001) with the occurrence of clinically affected pigs in the herd. Purchase of at least 5 pigs from herd SA was associated with an elevated risk (p less than 0.05) of isolation of toxigenic P. multocida.     

Response of the Rieger begonia to ozone and sulphur dioxide

Ozone (O₃) and sulphur dioxide (SO₂) are at present the two most pervasive air pollutants in the urban and suburban areas in which most Rieger begonias are grown. These air pollutants have been shown to impair crop productivity and quality in many other species. Controlled environment experiments were therefore conducted to elucidate the responses of ‘Schwabenland Red’ to separate or bombined O₃ and SO₂ at different growth stages.Plants at the early vegetative or prefloral stages had much more leaf damage after combined O₃ + SO₂ than after separate gases at 15 p.p.h.m. O₃ or 60 p.p.h.m. SO₂ for 5 days in a synergistic type of response. Exposure for an additional day at twice these concentrations greatly increased leaf damage. Growth reductions were not found after 2 weeks but delaying measurements until 8 weeks after treatment revealed some growth reductions.Experiments were conducted with flowering plants using O₃ of 0, 10, 20 or 30 p.p.h.m. in all combinations with SO₂ at 0, 60, 120 or 180 p.p.h.m. Fully developed leaves were damaged, and undamaged leaves were smaller and lighter on plants exposed to 20 p.p.h.m. or more O₃, 120 p.p.h.m. or more SO₂, or any combination of O₃ and SO₂ at or above 10 and 60 p.p.h.m., respectively.     

The effect of the molluscicide Frescon on smooth and cross-striated muscles of Lymnaea stagnalis and Helix aspersa

The molluscicide Frescon (N-(triphenylmethyl)morpholine) induces prolonged, irreversible contractions (contractures) in the isolated heart ventricle, penis retractor, and foot muscles of the aquatic snail Lymnaea stagnalis. It also causes contracture of the penis retractor muscle, but not the heart ventricle, of the terrestrial snail Helix aspersa. Since Frescon-induced contracture is not accompanied by a depolarization of the sarcolemma of Lymnaea heart ventricle muscle cells, it is suggested that the action of this molluscicide is associated with a membrane potential-independent activation of the contractile machinery. The rapid response of Lymnaea musculature to Frescon suggests that the direct contracture-inducing action of this molluscicide could explain its toxicity to the whole animal.