A model for gramicidin A′-phospholipid interactions in bilayers

A model is proposed for the effect of gramicidin A on the order and structure of phospholipid dispersions. According to this model, the addition of gramicidin A influences the surrounding lipids via two independent mechanisms. The first arises from a drop in surface pressure for those lipids substantially bounded by gramicidin A. The second mechanism arises from the increase in the phospholipid headgroup spacing due to the small polar region of the polypeptide. The model provides an explanation for the currently available NMR, X-ray diffraction and Langmuir monolayer results. The model also suggests mechanisms for the ability of gramicidin A to trigger a transition of the lipid from the lamellar to hexagonal II phase, the dependence of this transition on the lipid chain length and the formation of a lamellar phase with lysophosphatidylcholine.Abbreviations NMR nuclear magnetic resonance - DMPC dimyristoylphosphatidylcholine - S molecular order parameter - CSA chemical shift anisotropy - DPPC dipalmitoylphosphati-dylcholine - LPC lysophosphatidylcholine     

Phosphorus sorption and extractability in Andic soil incubated with plant residues of variable P content

Sodium acetate extractable P and P sorption were measured in a high P-sorbing Andept soil (Mission series) following incubation with Winter wheat (Triticum aestivum (L.)Nugaines) and red clover (Trifolium pratense (L.)Florex) plant residue. The effects of plant residue P content, moisture regime, and quantity added were evaluated over a twelve week incubation period. Increased extractable P and decreased P sorption occured when the P content of residue exceeded 0.1% P for both winter wheat and red clover. The extractable P decreased and the P sorbed increased for all treatments and the check as the incubation time progressed. Extractable P increased and P sorption decreased more under air dry moisture conditions than at –0.03 MPa or saturation. Extractable P levels increased and P sorption decreased with increasing amounts of added residue. The effects of the residue additions decreased with increasing incubation time.     

Purification and properties of a F420-nonreactive,membrane-bound hydrogenase from Methanosarcina strain Gö1

The distribution of the F₄₂₀-reactive and F₄₂₀-nonreactive hydrogenases from the methylotrophic Methanosarcina strain Gö1 indicated a membrane association of the F₄₂₀-nonreactive enzyme. The membrane-bound F₄₂₀-nonreactive hydrogenase was purified 42-fold to electrophoretic homogeneity with a yield of 26.7%. The enzyme had a specific activity of 359 mol H₂ oxidized · min^-1 · mg protein^-1. The purification procedure involved dispersion of the membrane fraction with the detergent Chaps followed by anion exchange, hydrophobic and hydroxylapatite chromatography. The aerobically prepared enzyme had to be reactivated anaerobically. Maximal activity was observed at 80°C. The molecular mass as determined by native gel electrophoresis and gel filtration was 77000 and 79000, respectively. SDS gel electrophoresis revealed two polypeptides with molecular masses of 60000 and 40000 indicating a 1:1 stoichiometry. The purified enzyme contained 13.3 mol S^2-, 15.1 mol Fe and 0.8 mol Ni/mol enzyme. Flavins were not detected. The amino acid sequence of the N-termini of the subunits showed a higher degree of homology to cubacterial uptake-hydrogenases than to F₄₂₀-dependent hydrogenases from other methanogenic bacteria. The physiological function of the F₄₂₀-nonreactive hydrogenase from Methanosarcina strain Gö1 is discussed.Abbreviations transmembrane electrochemical gradient of H^- - CoM-SH 2-mercaptoethanesulfonate - F₄₂₀ (N-l-lactyl--l-glutamyl)-l-glutamic acid phospodiester of 7,8-didemethyl-8-hydroxy-5-deazariboflavin-5-phosphate - F₄₂₀H₂ reduced F₄₂₀ - HTP-SH 7-mercaptoheptanoylthreonine phosphate - Mb. Methanobacterium - PMSF phenylmethyl-sulfonylfluoride - Cl₃AcOH trichloroacetic acid     

Morphogenesis of the foetal membranes and placenta of the root-rat (Tachyoryctes splendens (Rüppel))

The morphogenesis of the foetal membranes and placenta of the root-rat Tachyoryctes splendens was studied by light microscopy. Implantation of the blastocyst is antimesometrial. The mode of implantation is secondary interstitial and the decidual reaction is similar to that in other rodents. Amniogenesis is by cavitation with a temporary open epamniotic cavity. Inversion of the yolk sac is complete but the disappearance of the parietal segment occurs relatively late. With the breakdown of both the decidua capsularis and parietalis, the yolk sac wall near the margins of the placenta becomes bathed in a pool of maternal blood and tissue debris resembling the haemophagous organ. The yolk sac villi are well vascularized. The allantoic cavity is persistent up to the limb bud stage. The definitive placenta is haemochorial and shows conspicuous endodermal sinuses or placental pits.     

Platinum-supported Bilayer Lipid Membranes modified by ferrocene and its derivatives

The biferrocene-containing Schiff base complexes (1) and (2) were synthesized and characterized by elemental analyses and spectral data. The Pt-supported Bilayer Lipid Membranes (BLMs) modified by ferrocene and its derivatives were studied by cyclic voltametry (CV) and the electrochemical properties of this system are reported. The oxidation mechanism of electrocatalysis of ascorbic acid on the Pt-supported BLMs is discussed.     

Complex subcellular distributions of enzymatic markers in intestinal epithelial cells

Summary Current procedures for isolating intestinal epithelial cell surface and intracellular membranes are based on the assumption that each organelle is marked by some unique constitutent. This assumption seemed inconsistent with the dynamic picture of subcellular organization emerging from studies of membrane turnover and recycling. Therefore, we have designed an alternative fractionation which is independent ofa priori marker assignments. We subjected mucosal homogenates to a sequence of separations based on sedimentation coefficient, equilibrium density, and partitioning in aqueous polymer twophase systems. The resulting distributions of protein and enzymatic markers define a total of 17 physically and biochemically distinct membrane populations. Among these are: basal-lateral membranes, with Na,K-ATPase enriched 21-fold; brush-border membranes, with alkaline phosphatase enriched as much as 38-fold; two populations apparently derived from the endoplasmic reticulum; a series of five populations believed to have been derived from the Golgi complex; and a series of five acid phosphatase-rich populations which we cannot identify unequivocally. Each of the five enzymatic markers we have followed is associated with a multiplicity of membrane populations. Basallateral, endoplasmic reticulum, and Golgi membranes contain alkaline phosphatase at the same specific activity as the initial homogenate. Similarly, Na,K-ATPase appears to be associated branes at specific activities two-to seven-fold that of the initial homogenate.     

Quantity and quality of seston in an impounded and a free-flowing river in Virginia,U.S.A.

Seston in the impounded North Anna River (NAR) was analyzed and compared to that in the free-flowing South Anna River (SAR) in Virginia, U.S.A. A wet filtration technique was used to separate seston into five size classes. The overall quantity of organic seston was much lower in the NAR than in the SAR. The seston in the NAR was composed of more living organisms, in particular zooplankters in the medium large (234–864 µm) and small (105–234 µm) size classes and diatoms and other algae in the fine (43–105 µm) and very fine (25–43 µm) size classes. The percentage of zooplankton declined sharply before reaching the downstream study site (32 km). The seston in the NAR tended to consist of slightly larger particles, but 80–85% of the seston in both rivers was in the ultrafine (0.45–25 µm) size class, which was almost entirely composed of detritus. One indication of seston food quality, the organic/inorganic ratio, was considerably higher for the medium large and small size classes in the NAR immediately below the dam and for the fine and very fine size classes farther downstream. Another indication of food quality, the usable caloric content, showed that the seston in the NAR contained considerably less total assimilable energy as far as 32 km downstream from the dam, but that the medium large and small size classes were rich in assimilable energy immediately below the dam because of zooplankton released from the reservoir. Thus, the overall quantity and quality of seston was lower in the NAR, but the quality of the seston, for at least a short distance below the dam, was higher in a well-defined size range that can be effectively utilized by certain filter feeders.     

Sulphate reduction in oxic and sub-oxic North-East Atlantic sediments

Abstract Oxic and sub-oxic N.-E. Atlantic sediments were examined for sulphate-reducing activity. Oxygen and/or nitrate reduction are probably the dominant mineralisation processes in the abyssal plain sediment studied. A low rate of sulphate reduction (0.1 nmol SO²⁻₄/ml/day) was recorded in the surface 5 cm of the continental slope sediment, together with the presence of a range of sulphate-reducing bacteria (SRB). A higher activity of sulphate reduction (2.2 nmol SO²⁻₄/ml/day) occurred in the continental shelf sediment which led to a small decrease in pore water sulphate and an increase in titration alkalinity. This sediment contained approx. 10²–10³ acetate, lactate and propionate oxidising SRB/ml. No low- M _r organic acids were detected in these sediments. However, amendment with 75 μM acetate stimulated sulphate-reducing activity in the shelf sediment.     

The formation and biotransformation of cysteine conjugates of halogenated ethylenes by rabbit renal tubules

The nephrotoxicity of chlorotrifluoroethylene (CTFE) was examined using isolated rabbit renal tubules suspensions. Exposure of the tubules to CTFE resulted in consumption of CTFE, formation of a glutathione conjugate and inhibition of active organic acid transport. Synthetic cysteine, N-acetylcysteine or glutathione conjugates of CTFE inhibited transport indicating S-conjugation as a possible toxic pathway. 1,2-dichlorovinyl glutathione (DCVG), a model synthetic glutathione conjugate, was used to examine the degradation and toxicity of these conjugates. DCVG inhibited rabbit renal tubule transport in vivo and in vitro. The DCVG was found to be degraded with the evolution of glutamine and glycine to produce the ultimate nephrotoxicant, dichlorovinyl cysteine. Dichlorovinyl cysteine is then bioactivated with the release of ammonia. This sequential degradation explains the latency of DCVG-induced renal transport inhibition relative to dichlorovinyl cysteine. It is now evident that certain halogenated ethylenes are capable of being biotransformed to glutathione conjugates in the kidney with their subsequent hydrolysis to nephrotoxic cysteine conjugates.