Thermodynamic resolution of the iron-sulfur centers of the succinic dehydrogenase of Rhodopseudomonas sphaeroides.

A single alkaline wash removes most of the succinic dehydrogenase activity from chromatophores of Rhodopseudomonas sphaeroides. Three iron-sulfur centers are also removed by this washing. Two of these are ferredoxin-like centers with electron paramagnetic resonance signals at g_v = 1.94 and midpoint potentials of +50 and ?250 mV at pH 7. The third is a high-potential iron-sulfur protein type signal centered at g 2.01 and a midpoint potential of +80 mV at pH 7. These centers have very similar properties to those of the well-characterized mammalian succinic dehydrogenase and account for the majority of iron-sulfur centers observed in chromatophores. Because it is so easily removed, it is concluded that succinic dehydrogenase is located on the outer surface of the chromatophore membrane, a conclusion supported by the fact that removal of the enzyme does not interfere with the kinetics of light-induced electron flow, nor does it allow cytochrome c₂ to escape from inside the chromatophore vesicles.     

Identification and breeding biology of the diving petrels Pelecanoides georgicus and P. urinatrix exsul at South Georgia

Abstract

The biology of two species of diving petrel (Pelecanoides georgicus and P. urinatrix exsul) was studied at Bird Island, South Georgia. Existing criteria (bill shape and morphology, wing length) for distinguishing these species are reviewed, and several new characters are recognised. For adults bill depth, the colour of the posterior part of the tarsus, and vocalisations are distinctive; chicks have down of different colours (pale grey in P. u. exsul, dark grey in P. georgicus). At South Georgia the species breed in different habitats and at different times - P. u. exsul nests in steep tussock slopes in very early summer, P. georgicus in fine scree slopes in midsummer. P. georgicus lays a proportionately larger egg, and has an incubation period of 46 days (c.54 days in P. u. exsul) and a chick fledging period of 46 days. The fledging period of P. u. exsul is 54 days, very similar to recorded values for P. u. urinatrix (53.5 days) and P. u. chathamensis (56 days). Data on feeding frequency and feed size were derived from daily chick weighings and from twice-daily weighing during 30 days preceding fledging. In both species chicks are fed every night, and often by both parents. In P. georgicus true mean chick feed size is c.37 g; in P. u. exsul it may be slightly less. Analysis of chick stomach contents suggests that P. u. exsul feeds extensively on copepods, whereas P. georgicus largely takes krill (Euphausia superba). P. u. exsul breeding adults commence moulting before their chicks have fledged; P. georgicus moults exclusively in the non-breeding season. Ectoparasites were collected, and the feather louse Pelmatocerandra setosa was found to be restricted to P. u. exsul, and Pelmatocerandra enderleini to P. georgicus. P. georgicus, which breeds later and whose chicks fledge faster, is suggested to be better adapted to the climatic and marine environmental conditions than P. u. exsul, which may be the more recent colonist of these high latitudes.     

Long-distance dispersal of the beach strawberry,Fragaria chiloensis,from North America to Chile and Hawaii

Background and AimsThe beach strawberry, Fragaria chiloensis, is found in a narrow coastal band from the Aleutian Islands to central California and then jumps thousands of kilometres all the way to Hawaii and Chile. As it probably had a North American origin, it must have been introduced to the other locations by long-distance dispersal. The aim of this study was to determine which agent carried the beach strawberry to its Pacific and South American locations.MethodsA deductive framework was constructed to separate between the possible modes of long-distance dispersal involving animals, wind and ocean currents. Bird migration was subsequently identified as the most likely scenario, and then the routes, habitats, feeding preferences and flight distances of all the shorebird species were evaluated to determine the most likely carrier.Key ResultsSix species migrate between North America and Chile and feed on the beaches and rocky shores where F. chiloensis grows naturally: Black-bellied Plovers, Greater Yellowlegs, Ruddy Turnstones, Sanderlings, Whimbrels and Willets. Of these, only two eat fruit and migrate in long continuous flight: Ruddy Turnstones and Whimbrels. Two species travel between North America and Hawaii, eat fruit and forage on the beaches and rocky shores where F. chiloensis grows naturally: Pacific Golden-plovers and Ruddy Turnstones. Ruddy Turnstones eat far less fruit than Pacific Golden-plovers and Whimbrels, making them less likely to have introduced the beach strawberry to either location.ConclusionsWe provide evidence that F. chiloesis seeds were probably dispersed to Hawaii by Pacific Golden-plovers and to Chile by Whimbrels.     

Quality assurance of monoclonal products: Virologic and molecular biologic considerations

Summary The FDA has set limits concerning the viral and molecular contamination of monoclonal antibody products intended for human use. Industry has an obligation to be as familiar with these limits as it has been with federal requirements pertaining to pyrogens and bacteria. The assessment of risk from polynucleotides, based on molecular biologic and existing technical limitations, is discssed, as is the strategy of validating the purification of monoclonal antibodies of viral contaminants in terms of an indicator organism concept.     

Spectroscopic properties of the hydroxylase of methane monooxygenase

The hydroxylase component of methane monooxygenase (EC 1.14.13.25), which catalyzes the oxidation of methane to methanol, has been studied by visible, electron spin resonance and X-ray spectroscopies. The enzyme appears to possess a mu-oxo- or mu-hydroxo-bridged binuclear iron site, with no sulfur ligands to the cluster. Each Fe has 4-6 oxygen (or nitrogen) ligands, at an average distance of 1.92 +/- 0.03 A. The Fe-Fe distance is 3.05 +/- 0.05 A. Essentially all of the irons are in the Fe3+ state as the enzyme is prepared, but reduction with N-methylphenazonium methosulfate generates ESR-detectable states that appear to emanate from mixed-valence binuclear sites. One of these, with gav near 1.85, displays typical Curie law microwave saturation behavior, but the other, gav near 1.73, has a very potent method of spin-relaxation. Together they account for approximately 0.6 spins per molecule.     

The role of the Rieske iron-sulfur center as the electron donor to ferricytochrome c2 in Rhodopseudomonas sphaeroides

The Rieske iron-sulfur center in the photosynthetic bacterium Rhodopseudomonas sphaeroides appears to be the direct electron donor to ferricytochrome c₂, reducing the cytochrome on a submillisecond timescale which is slower than the rapid phase of cytochrome oxidation ($\text{t}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{3–5 μ}\text{s}$). The reduction of the ferricytochrome by the Rieske center is inhibited by 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole (UHDBT) but not by antimycin. The slower (1–2 ms) antimycin-sensitive phase of ferricytochrome c₂ reduction, attributed to a specific ubiquinone-10 molecule (Q_z), and the associated carotenoid spectral response to membrane potential formation are also inhibited by UHDBT. Since the light-induced oxidation of the Rieske center is only observed in the presence of antimycin, it seems likely that the reduced form of Q_z (Q_zH₂) reduces the Rieske center in an antimycin-sensitive reaction. From the extent of the UHDBT-sensitive ferricytochrome c₂ reduction we estimate that there are 0.7 Rieske iron-sulfur centers per reaction center.UHDBT shifts the EPR derivative absorption spectrum of the Rieske center from g_y 1.90 to g_y 1.89, and shifts the E_m,7 from 280 to 350 mV. While this latter shift may account for the subsequent failure of the iron-sulfur center to reduce ferricytochrome c₂, it is not clear how this can explain the other effects of the inhibitor, such as the prevention of cytochrome b reduction and the elimination of the uptake of H⁺_II; these may reflect additional sites of action of the inhibitor.     

A microcosm for measuring survival and conjugation of genetically engineered bacteria in rhizosphere environments

A microcosm is described to evaluate and measure bacterial conjugation in the rhizosphere of barley and radish with strains ofPseudomonas cepacia. The purpose was to describe a standard method useful for evaluating the propensity of genetically engineered microorganisms (GEMs) to transfer DNA to recipient bacteria. Results demonstrated the formation of transconjugants from the rhizosphere of each plant 24 h after inoculation. Transconjugant populations peaked at 1.8 × 10² colony forming units (CFU)/g root and associated soil in barley and 2.0×10² CFU/g root and associated soil in radish; they then declined over the next five days of the experiment. No significant differences were found in the survival of transconjugant populations monitored from the two plant species. The microcosm was also used to document the formation of false positive transconjugants, which resulted from donor and recipientP. cepacia mating on the surface of selective agar plates instead of in microcosms. Transconjugants resulting from such plate mating occurred in substantial numbers during the first 5 days of the experiment but declined to undetectable numbers by day 7. The use of nalidixic acid was investigated to determine the magnitude of plate mating. The number of transconjugants detected from radish rhizosphere was reduced by two orders of magnitude by including nalidixic acid in the plating medium; this indicated that 99% of the transconjugants were a result of plate mating.     

Is the cognitive function of older patients affected by antihypertensive treatment? Results from 54 months of the Medical Research Council's trial of hypertension in older adults.

OBJECTIVE--To establish whether initiation of treatment with diuretic or beta blocker is associated over 54 months with change in cognitive function. DESIGN--A cognitive substudy, nested within a randomised, placebo controlled, single blind trial. SETTING--226 general practices from the Medical Research Council''s general practice research framework. SUBJECTS--A subset of 2584 subjects sequentially recruited from among the 4396 participants aged 65-74 in the trial of treatment of hypertension in older adults. The 4396 subjects were randomised to receive diuretic, beta blocker, or placebo. Subjects had mean systolic pressures of 160-209 mm Hg and mean diastolic pressures <115 mm Hg during an eight week run in. OUTCOME MEASURES--The rate of change in paired associate learning test (PALT) and trail making test part A (TMT) scores (administered at entry and at 1, 9, 21, and 54 months) over time. RESULTS--There was no difference in the mean learning test coefficients (rate of change of score over time) between the three treatments: diuretic -0.31 (95% confidence interval -0.23 to -0.39), beta blocker -0.33 (-0.25 to -0.41), placebo -0.30, (-0.24 to -0.36). There was also no difference in the mean trail making coefficients (rate of change in time taken to complete over time) between the three groups: diuretic -2.73 (95% confidence interval -3.57 to -1.88), beta blocker -2.08 (-3.29 to -0.87), placebo -3.01, (-3.69 to -2.32). A less conservative protocol analysis confirmed this negative finding. CONCLUSION--Treating moderate hypertension in older people is unlikely to influence, for better or for worse, subsequent cognitive function.