Effect of straw on the behaviour of growing pigs

The effects of straw on the behaviour of young growing pigs were studied in two experiments. In Experiment 1, groups of three pigs, aged about 7 weeks, were housed in raised decks with or without straw bedding. Time-lapse video recording showed no major differences between treatments in the amount or daily pattern of overall activity and feeding. Detailed observations showed that bedding reduced the incidence of rooting and chewing on pen-mates, but had little effect on other social activities such as mounting and aggressive biting. Experiment 2 compared groups of eight growing pigs aged about 10 weeks, housed in floor pens with or without a small amount of straw provided daily in a rack. Pigs with straw concentrated more of their daily activity into the period when straw was fresh, but the total amount of overall activity and time spent feeding were not affected. Rooting and chewing of pen-mates were the only social activities reduced by the provision of fresh straw. In these studies, where straw was not required to compensate for deficiencies such as low temperatures or hunger, the one major function of straw was to provide a stimulus and outlet for rooting and chewing, with a resulting reduction in such activities directed at pen-mates.     

Growth of an adherent continuous cell line entrapped in a peg/alginate matrix

Summary CHO-K1 cells, an anchorage-dependent line, were entrapped in beads prepared from a Na alginate/polyethylene glycol mixture and grown, through successive passages, to an average maximum density of 4.5×10⁷ viable cells/g of bead. Cell growth and viability was unaffected by repeated alginate re-solubilization and reformation of the gel beads through five passages.     

Dictyostelium discoideum lipids modulate cell-cell cohesion and cyclic AMP signaling.

During Dictyostelium discoideum development, cell-cell communication is mediated through cyclic AMP (cAMP)-induced cAMP synthesis and secretion (cAMP signaling) and cell-cell contact. Cell-cell contact elicits cAMP secretion and modulates the magnitude of a subsequent cAMP signaling response (D. R. Fontana and P. L. Price, Differentiation 41:184-192, 1989), demonstrating that cell-cell contact and cAMP signaling are not independent events. To identify components involved in the contact-mediated modulation of cAMP signaling, amoebal membranes were added to aggregation-competent amoebae in suspension. The membranes from aggregation-competent amoebae inhibited cAMP signaling at all concentrations tested, while the membranes from vegetative amoebae exhibited a concentration-dependent enhancement or inhibition of cAMP signaling. Membrane lipids inhibited cAMP signaling at all concentrations tested. The lipids abolished cAMP signaling by blocking cAMP-induced adenylyl cyclase activation. The membrane lipids also inhibited amoeba-amoeba cohesion at concentrations comparable to those which inhibited cAMP signaling. The phospholipids and neutral lipids decreased cohesion and inhibited the cAMP signaling response. The glycolipid/sulfolipid fraction enhanced cohesion and cAMP signaling. Caffeine, a known inhibitor of cAMP-induced adenylyl cyclase activation, inhibited amoeba-amoeba cohesion. These studies demonstrate that endogenous lipids are capable of modulating amoeba-amoeba cohesion and cAMP-induced activation of the adenylyl cyclase. These results suggest that cohesion may modulate cAMP-induced adenylyl cyclase activation. Because the complete elimination of cohesion is accompanied by the complete elimination of cAMP signaling, these results further suggest that cohesion may be necessary for cAMP-induced adenylyl cyclase activation in D. discoideum.     

Cloning and sequencing of a gene encoding an aminoglycoside 6'-N-acetyltransferase from an R factor of Citrobacter diversus.

The aacA1 gene, which encodes a 6'-N-acetyltransferase [AAC(6')-I] mediating resistance to kanamycin, tobramycin, and amikacin, was cloned from the Citrobacter diversus R plasmid pBWH100 into the Escherichia coli vector pBR322. The complete nucleotide sequence of the gene and flanking regions was determined. A protein of approximately 21 kilodaltons was identified when the chimeric plasmid encoding the aacA1 gene was introduced into E. coli maxicells. This value is consistent with the size predicted for a protein translated from the open reading frame of the gene.     

Influence of molecular packing and phospholipid type on rates of cholesterol exchange

The rates of [14C]cholesterol transfer from small unilamellar vesicles containing cholesterol dissolved in bilayers of different phospholipids have been determined to examine the influence of phospholipid-cholesterol interactions on the rate of cholesterol desorption from the lipid-water interface. The phospholipids included unsaturated phosphatidylcholines (PC's) (egg PC, dioleoyl-PC, and soybean PC), saturated PC (dimyristoyl-PC and dipalmitoyl-PC), and sphingomyelins (SM's) (egg SM, bovine brain SM, and N-palmitoyl-SM). At 37 degrees C, for vesicles containing 10 mol% cholesterol, the half-times for exchange are about 1, 13, and 80 h, respectively, for unsaturated PC, saturated PC, and SM. In order to probe how differences in molecular packing in the bilayers cause the rate constants for cholesterol desorption to be in the order unsaturated PC greater than saturated PC greater than SM, nuclear magnetic resonance (NMR) and monolayer methods were used to evaluate the cholesterol physical state and interactions with phospholipid. The NMR relaxation parameters for [4-13C]cholesterol reveal no differences in molecular dynamics in the above bilayers. Surface pressure (pi)-molecular area isotherms for mixed monolayers of cholesterol and the above phospholipids reveal that SM lateral packing density is greater than that of the PC with the same acyl chain saturation and length (e.g., at pi = 5 mN/m, where both monolayers are in the same physical state, dipalmitoyl-PC and palmitoyl-SM occupy 87 and 81 A2/molecule, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of the structure of the lipid-water interface on the activity of hepatic lipase

Factors affecting the hydrolytic activity of purified rat hepatic lipase have been examined in mixed-monolayer systems. When nonsubstrate lipids [either egg sphingomyelin or beta-O-hexadecyl-gamma-O-(1-ocadec-9-enyl)-DL-phosphatidylcholine (OPPC-ether)] were used as inert matrices, hydrolytic activity for both triolein and dioleoylphosphatidylethanolamine was shown to decrease with increasing surface pressure (pi); negligible activity occurred at pi greater than or equal to 30 mN/m. Examination of the effect of introduction of cholesterol into either matrix containing 2 mol % triolein indicated that the mean molecular area decreased with increasing cholesterol and that, at pi = 24 mN/m, triolein was fully miscible in the sphingomyelin matrix at cholesterol concentrations less than or equal to 32.5 mol % and in the OPPC-ether matrix at cholesterol concentrations less than or equal to 49 mol %. Above these critical concentrations of cholesterol, the phase diagrams indicate transitions that suggest that triolein is forced out of the monolayer. Introduction of increasing amounts of cholesterol into either inert matrix increased the rate of hydrolysis of triolein by hepatic lipase, although by different degrees. There are at least two factors contributing to these effects: (1) condensation of the monolayer by cholesterol, thus increasing the total surface concentration of triolein at pi = 24 mN/m in the constant area surface balance, and (2) some change in triolein conformation and/or accessibility since at identical surface concentrations of triolein (8.7 +/- 0.1 pmol/cm2) and pi (24 mN/m) the rate of hydrolysis of triolein by hepatic lipase is 1.5-fold higher in the OPPC-ether matrix than in the egg sphingomyelin matrix.(ABSTRACT TRUNCATED AT 250 WORDS)     

Polymerization of G-actin by myosin subfragment 1

The polymerization of actin from rabbit skeletal muscle by myosin subfragment 1 (S-1) from the same source was studied in the depolymerizing G-actin buffer. The polymerization reactions were monitored in light-scattering experiments over a wide range of actin/S-1 molar rations. In contrast to the well resolved nucleation-elongation steps of actin assembly by KC1 and Mg2+, the association of actin in the presence of S-1 did not reveal any lag in the polymerization reaction. Light scattering titrations of actin with S-1 and vice versa showed saturation of the polymerization reaction at stoichiometric 1:1 ratios of actin to S-1. Ultracentrifugation experiments confirmed that only stoichiometric amounts of actin were incorporated into a 1:1 acto-S-1 polymer even at high actin/S-1 ratios. These polymers were indistinguishable from standard complexes of S-1 with F-actin as judged by electron microscopy, light scattering measurements, and fluorescence changes observed while using actin covalently labeled with N-(1-pyrenyl)iodoacetamide. F-actin obtained by polymerization of G-actin by S-1 could initiate rapid assembly of G-actin in the presence of 10 mM KC1 and 0.5 mM MgCl2 and showed normal activation of MgATPase hydrolysis by myosin.     

The influence of the triglyceride content of low density lipoprotein on the interaction of apolipoprotein B-100 with cells

To study the effect of triglyceride content of low density lipoprotein (LDL) on its physicochemical and biological properties, we have depleted the triglyceride by incubation with hepatic lipase (HL-LDL) and raised the triglyceride by incubation of HL-LDL with very low density lipoprotein and lipoprotein-deficient serum. HL-LDL was taken up by human monocyte-derived macrophages and by human skin fibroblasts at an increased rate compared to untreated LDL. Incubation of the various LDL preparations revealed that cellular LDL degradation as well as LDL-mediated cholesterol esterification were inversely related to the triglyceride content of the LDL preparation. Modification of the triglyceride content of LDL also was associated with changes in the free fatty acid content, but the interaction of the LDL with cells was unaffected by the level of this component. The triglyceride content of LDL was found to be reciprocally related to the number of free lysine amino groups of LDL apolipoprotein B (apoB) which could be labeled with trinitrobenzenesulfonic acid. 13C-Nuclear magnetic resonance (NMR) spectra of native LDL and HL-LDL samples containing [13CH3]2 lysine residues formed by reductive methylation (11-13% modification) showed that the arrangement of apoB lysines is perturbed by the exposure to hepatic lipase. The ratio of labeled lysines with pK 8.9 to those with pK 10.5 exposed on the surface of LDL particles was decreased by about 40% by lipase treatment. These effects are apparently due to changes in local apoB conformation because circular dichroism spectra revealed that the average secondary structure of the entire apoB molecule is the same in native LDL and HL-LDL. The triglyceride content of LDL reciprocally affected its binding to a monoclonal antibody which recognizes epitopes around the LDL receptor binding domain of apoB. The above evidence indicates that modulation of the core triglyceride and possibly also surface phospholipid content of LDL can alter the conformation of apoB on the surface of the particle, thereby influencing the interaction with cell surface LDL receptors.     

Preparation of cytoplasmic bodies (cytospheres) from isolated hepatocytes and their biochemical properties

The controlled centrifugation of isolated rat hepatocytes at 260 000 g results in the formation of membrane-bounded cell fragments that we have termed 'cytospheres'. A method is described for the isolation of these cytospheres. Cytospheres are spherical, have a mean diameter of 9.2 +/- 3.2 microns (SD) and a protein content of 225 +/- 12 mg/g wet wt. About 3% of the protein from the original isolated hepatocyte suspension is recoverable. Transmission electron microscopy (TEM) shows cytospheres to possess a trilaminar membrane, and a finely granular hyaloplasm generally devoid of organelles, filaments and microtubules. Freeze-fracture studies reveal a membrane structure typical of a plasma membrane. Ouabain and wheat germ agglutinin (WGA)-binding studies indicate that the original orientation of the plasma membrane is maintained throughout the formation of the cytospheres. The cytospheres have also been characterized biochemically. Cytospheres are enriched in the enzymes normally associated with the hyaloplasm, whereas the activities of enzymes localized in organelles are greatly diminished. Lipid analysis of the cytosphere membrane indicates that it is derived from the plasma membrane of the hepatocyte. Cytospheres are sensitive to changes in the osmolarity and ionic composition of their environment. Cytospheres should therefore prove a useful preparation for the study of hyaloplasm metabolism and of plasma membrane receptor and permeability properties.