Antimicrobial defense mechanisms in the horseshoe crab, Limulus polyphemus: preliminary observations with heat-derived extracts of Limulus amoebocyte lysate.

Heat-derived (60°C) extracts of Limulus amoebocyte lysate (LAL) were found to contain potent “broad-spectrum” antimicrobial activity. Additional heating of the LAL extracts to 100°C for 30 min completely inactivated the antimicrobial activity and served as a control. Antimicrobial activity was observed over a temperature range of 0° to 37°C (higher temperatures not tested) with greatest activity at 37°C. Antimicrobial activity of LAL extracts was variable when tested against Gram-negative bacteria of the family Enterobacteriaceae. A twofold concentration of the extracts resulted in a significant decrease in antimicrobial effectiveness. Dialysis of single- and double-strength LAL extracts against deionized water produced a marked and significant enhancement of antimicrobial activity against both resistant and sensitive species, confirming the presence of a dialyzable inhibitor(s). Dialyzed LAL extracts were active against 13 of 14 species of Enterobacteriaceae tested. Two strains of Pseudomonas aeruginosa were susceptible as were two of three Gram-positive cocci tested. Highly sensitive bacterial species were rapidly killed with a greater than 90% reduction in viable counts occurring within the first 30 min of reaction time. Dialyzed LAL extracts also possessed considerable antifungal activity. The role of the Limulus polyphemus amoebocyte in defense against microbial invasion and dissemination is discussed.     

Rat and human embryo and post-natal sera contain a potent endogenous competitor of estrogen-rat alpha-fetoprotein interactions

A highly active inhibitor of the binding of estrone and estradiol-17β to rat alpha-fetoprotein is demonstrated for the first time in embryo, immature and adult rat sera as well as in fetal and adult human sera. The competitive character and the narrow specificity of this inhibition effect is shown. The major compound responsible for this activity is isolated by successive column Sephadex LH₂₀ and thin layer chromatography : it is characterized as a nonpolar, nonphenolic, dialysable and thermostable substance, unreactive towards anti-estrone and anti-estradiol-17β anti-bodies. The possible biological role of an endogenous non-estrogen ligand of rodent fetoproteins is discussed.     

SEAWEED ANATOMY AND PHOTOSYNTHETIC PERFORMANCE: THE ECOLOGICAL SIGNIFICANCE OF LIGHT GUIGES,HETEROGENEOUS ABSORPTION AND MULTIPLE SCATTER12

Light absorption by two green seaweeds with similar photophysiology but different anatomies are compared: i) Ulva lactuca var. rigida (C. Ag.) Le Jolis, an optically translucent species of two cell layers both bearing chloroplasts; and, ii) Codium fragile subsp. tomentosoides (van Goor) Silva, an optically opaque species with a colorlelss medulla surrounded by a cortex of choloroplast-bearing utriclels. Thallus absorptance (fraction of incident light absorbed) was measured for various pigment contents. Absorptance by U. lactuca was dependent on pigment concentration in an exponential manner and never exceeded 0.6, whereas absorptance by C. fragile was independent of pigment concentration and always approached a balue of 1.0. Water in the medullary tissue of C. fragile is often of the utricles. The utricles appear to be “integrating spheres” enhancing the capture of incident light, aided by the wave-guide function of the thin peripheral layer of cytoplasm and a reflector function at their base. Photosynthitic performance for U. lactuca saturates at high light intensities and attenuates rapidly with decreasing intensities. In contrast, photosynthetic performance for C. fragile saturates at low light intensities and attenuates slowly with diminishing radiation. Extrapolated diel variation in photosynthesis shows that U. lactuca's anatomy is adaptive for high light intensity environments, whereas C. fragile's anatomy is adaptive for low light intensity environments. Both seaweeds fit into the ecological category of “fugitive” species, and compete in the Long Island Sound (Atlantic Ocean) rocky intertidal for free-space. Predictions are presented for relative species abundances along a monotonic gradient of light intensity.     

Kinetics of the cooperativity of the Ca2+-transporting adenosine triphosphatase of sarcoplasmic reticulum and the mechanism of the ATP interaction.

The extent of the negative cooperativity with MgATP of the Ca²⁺-stimulated ATPase activity of sarcoplasmic reticulum has been studied with various membrane preparations and under various conditions. Preparations studied were fragmented sarcoplasmic reticulum vesicles, deoxycholate-solubilized and fractionated ATPase, triton extracted reticulum, vesicles reconstituted from either detergent, and limited trypsin digests of the reticulum. Conditions studied were suboptimal, optimal, and inhibitory Ca²⁺ concentrations; temperatures from 13 to 46 °C; 1 or 5 mm MgCl₂; 0.1 m KCl, 0.1 m NaCl, or no added salt; and Triton or deoxycholate present in the assay. With preparations in which vesicles could accumulate Ca²⁺ ion, the ionophore A23187 was added to prevent inhibition by internal Ca²⁺ ions. Under all circumstances, the negative cooperativity of MgATP was present (Hill coefficient of 0.2 to 0.8), indicating the persistence of the properties of the enzyme molecule and its lipid environment giving rise to kinetic negative cooperativity. Attempts to measure the number of ATP sites by protection against N-ethylmaleimide inactivation and by binding of an analog suggested, but did not prove, that there was only one specific, active ATP binding site below 0.5 mm. These results are interpreted to be consistent with either of two mechanisms for ATP cooperativity of the Ca²⁺-stimulated ATPase activity of sarcoplasmic reticulum: (a) a single, high affinity ATP active site and a second, lower affinity “allosteric” activator site; or (b) a single ATP site which demonstrates two affinities through some kinetic mechanism such as a substrate-induced, slow transition.     

The Effects of Heat and Cobalt-60 Gamma-Radiation on Excystation of the Rat Coccidium,Eimeria nieschulzi Dieben, 1924*

SYNOPSIS. Sporulated oocysts of Eimeria nieschulzi Dieben, a rat coccidium, were exposed for 1 hr to Cobalt-60 γ-radiation (15, 30. or 60 k-rads), to heat (35, 40, or 45 C). or to both concurrently (15, 30, or 60 k-rads at 35 C) to compare the excystation capabilities of treated vs nontreated parasites. Intact, treated oocysts appeared structurally unaltered when viewed with the light microscope. Excystation of sporozoites occurred in all treated groups when their sporocysts were exposed to a trypsin-sodium taurocholate (TST) fluid, but after 150 min in TST the excystation rate was significantly lower than in non-treated sporocysts. Sporozoites which excysted from treated sporocysts were abnormal both in the excystation process and in their form and movement once outside the sporocyst.     

Effects of water conductivity on electrocommunication in the weak-electric fish Brienomyrus niger (Mormyriformes)

A characteristic electric organ discharge display in social encounters between mormyrid fish is a temporary discharge cessation. Using this response, we have investigated the useful range of electrocommunication under different water conductivity conditions in the mormyrid Brienomyrus niger. An individual fish was confined to a porous ceramic shelter tube and moved from a starting distance of 380 cm toward a similarly confined conspecific until discharge, cessation occurred. The moved fish was subsequently returned to its original, position. Water conductivity affects the peak-to-peak source voltage of the electric organ and the sensitivity of the fish's electroreceptors. Within a range of 10 to 36 000 μS/cm, the peak-to-peak amplitude of the electric organ discharge declined as a power function. At 120 μS/cm, the amplitude was 50%, and at 300μS/cm, 30% of the 10 μS/cm value. The interfish distance at which discharge cessation occurred and the associated electric field gradients were dependent on water conductivity and upon the spatial orientation of the two fish (end-to-end or parallel orientations of their shelter tubes). The respective ranges were from 135 cm and 0.02 mV/cm at 52 μS/cm (parallel orientation) to 22 cm and 0.36 mV/cm at 678 μS/cm (end-to-end orientation). When the data for both tube orientations were combined, the relationship between water conductivity (x) and the distance at which discharge cessation occurred (y) could be expressed by a power function, y=K·x^a (with K=10^2.97 and a=?0.56). When an electrically ‘silent’ fish was moved away from its conspecific, a discharge resumption in the form of a high-frequency rebound occasionally effected changes in the other fish's discharge activity at distances up to 157 cm (with an associated electric, field gradient of 0.01 mV/cm under the lowest conductivity condition).     

Light-dependent conformational change at rhodopsin's cytoplasmic surface detected by increased susceptibility to proteolysis

Rhodopsin in bovine photoreceptor disk membranes was subjected to limited proteolysis by thermolysin, removing twelve amino acids from rhodopsin's carboxyl terminus. (1) The rate of proteolysis is significantly faster with rhodopsin following exposure to light than with unbleached rhodopsin, provided that the incubation conditions (pH, temperature) favor the formation of metarhodopsin II. (2) If the disk membranes are illuminated under conditions in which metarhodopsin I is the predominant photoproduct (pH 8.5, 0°C), no increase in the rate of proteolysis is observed compared to unilluminated membranes. (3) The light-induced increase in the rate of proteolysis is transient: it slowly decays in the dark to the original rate found for unbleached rhodopsin. The enhanced susceptibility to proteolysis appears to measure a conformational change at rhodopsin's cytoplasmic surface which is first exhibited at the metarhodopsin II stage. This and possibly other light-dependent changes may allow rhodopsin to mediate its signal as a light-receptor protein by binding to and activating certain rod cell enzymes.     

Characterization and determination of titratable groups of proteins by linearization of titration curves. II. Application to lysozyme

The potentiometric acid-base titration curve of fully protonated lysozyme at ionic strengths of 0.10 and 1.0 m has been performed. The stoichiometry and the pK_a values of each titratable group have been determined through the linearization of titration curves. Two types of carboxylic groups with pK_a values of 3.76 and 5.02, the imidazole group with pK_a 7.37 and the amine group with pK_a 9.63, have been identified at an ionic strength of 0.10 m at 25.0°C. The number of titratable groups found per mole of protein has been 5.12 and 5.60 for the two types of carboxylic groups, 1.13 for the imidazole group, and 3.19 for the amino groups. The endpoint of the titration of the protein obtained by this method accords quite well with the endpoint obtained by the use of Gran function applied to the excess of strong base.     

阿胶,新阿胶,黄明胶,马皮胶,杂皮胶的质量对比研究：Ⅰ.氨基酸分析

本文用835-50型氨基酸自动分析仪测定了阿胶、新阿胶、黄明胶、马皮胶和杂皮胶中游离氨基酸和总氨基酸的含量,并计算出游离氨基酸、总氨基酸按侧链基团分类和按医药用途分类的相对含量。结果表明,各种氨基酸含量一般以驴皮胶为较高,但与其他几种胶比较并无显著差异,似可以猪皮马皮、牛皮代替驴皮。这也说明,单从氨基酸的含量来鉴别胶的种类和质量似乎不可行。     

Non-light-dependent Shikimate Pathway in Plastids from Pea Roots

Non-green plastids (leucoplasts) isolated from pea roots are shown to be considerably active in forming aromatic amino acids by the shikimate pathway which, in contrast to the chloroplast pathway, is independent of light. Supply of phosphoenolpyruvate and 3-dehydroquinate, 3-dehydroshikimate, shikimate and quinate effectively enhances the formation of aromatic amino acids suggesting an intra- or/and intercellular intermediate transport.