The Effect of Sterilization, pH, Filler and Spore Inoculum Concentration on the Preparation of Alginate Pellets

Alginate encapsulation of an atoxigenic strain of Aspergillus flavus was studied in order to optimize encapsulation of fungal inocula with alginic acid. Sterilization by autoclaving is known to depolymerize sodium alginate. Buffered solutions (pH = 7-8) reduced this effect. Autoclaving the alginate solution with a filler/nutrient further inhibited the depolymerization reaction. Autoclaving under optimal conditions allowed a less expensive alginate (medium viscosity) to be used at a lower concentration (1%) to produce a stable product. The lowest cost pellets resulted from use of 1% medium viscosity sodium alginate with 10% cotton-seed meal. Further savings may be achieved by performing fermentations directly in alginate-nutrient mixtures and thus eliminating the mixing and blending steps. In such formulations, the nutrient composition and length of fermentation must be adjusted to prevent alginate hydrolysis. The ultimate composition of alginate pellets is influenced by the diffusion of nutrients during gelation. Up to 65% of water-soluble nutrients were lost from alginate pellets during gelation. Once pellets are introduced into the environment, organisms other than the formulated agent compete for pelleted nutrients. A minimum concentration of the biocontrol agent must be present to ensure the agent excludes competitors and successfully converts the nutrients to biomass. For A. flavus, 5000 spores g^-1 were required.     

Female sand gobies prefer good fathers over dominant males

Female preference for males successful in male-male competition is generally assumed to result in mating with high quality males. Here I report results from an experiment disentangling the effects of intra- and intersexual selection in the sand goby, Pomatoschistus minutus, a marine fish that exhibits paternal care. I show that large males are successful in male–male competition, but contrary to what one would expect, dominants are not preferred by females and are not better at taking care of the eggs. Female preference, however, correlated with the subsequent hatching success of the eggs. Thus, female choice selects for good parenting. Hence, direct benefits in the form of superior paternal care can explain female choice in this species, supporting a good parent process of sexual selection. However, choosing on the outcome of male–male competition does not enable females to mate with the ''best'' males.     

Formulating Atoxigenic Aspergillus flavus for Field Release

A procedure was developed to encapsulate mycelia of an atoxigenic strain of Aspergillus flavus in alginate pellets for seeding into agricultural fields in order to reduce aflatoxin contamination via competitive exclusion. Kaolin, a clay filler commonly employed in alginate formulations, was detrimental to pellet performance as measured by spore yield. Corn cob grits, a by-product of the corn industry, was found to be an excellent replacement for kaolin. Of nine nutritive adjuvants tested, wheat gluten improved pellet performance the most, although gluten concentrations above 5% were difficult to process. The best formulation tested consisted of 1% sodium alginate, 5% corn cob grits and 5% wheat gluten. On a 'per gram' basis, this alginate formulation yielded more spores than either A. flavus sclerotia or colonized wheat seed. Pesticides were also tested as adjuvants with potential use for protecting pellets under field conditions. Only one (chloramphenicol) of four tested pesticides (the others were dichloran, rose Bengal and cyfluthrin) reduced pellet sporulation. Formulations with or without pesticide adjuvants retained similar spore yield potential during a 2-year storage at 8 C. However, spore production in stored products lagged behind that of fresh products. At 75% relative humidity (RH), pellet storage stability decreased with increasing temperature from 27 to 42 C. Pellet spore yield at 32 C decreased as RH decreased from 100 to 90%. Sporulation occurred at 90% RH but not at 88% RH. Spore yield varied widely in four field tests, and the cumulative spore yield was inversely correlated (r2= -0.798, P 0.01) with rainfall. The results suggest that alginate pellets may be effective formulations for delivery of atoxigenic A. flavus strains to furrow-irrigated cotton in desert environments, where aflatoxin contamination of cottonseed is most severe.     

Synthesis and bioassay of isoprenoid 3-alkylthio-1,1,1-trifluoro-2-propanones: Potent,selective inhibitors of juvenile hormone esterase

Four 3-alkylthio-1,1,1-trifluoro-2-propanones with juvenile hormone-like side chains were prepared from citronellol and homogeraniol. These substrates were designed as possible transition-state analogs for the juvenile hormone (JH)-specific esterases present in insects. These four isoprenoid trifluoromethyl ketones were assayed in vitro with JH esterase and general esterases from larvae of the cabbage looper, Trichoplusia ni (Lepidoptera, Noctuidae), and with eel acetylcholinesterase and bovine chymotrypsin. JH esterase inhibition I₅₀ values were in the nanomolar range for all four compounds, while the other esterases had I₅₀'S which were 10³ to 10⁵ higher. The high selectivity of these inhibitors is believed to be due to their similarity in size and functionality to natural JH III. Treatment of T. ni larvae in vivo with solutions of the most active analog, 3-[(E)-4,8-dimethyl-3,7-nonadienylthio]-1,1,1-trifluoro-2-propanone (DNTFP) causes a dose-dependent delay in pupation and a concurrent selective inhibition of JH esterase. These data support the hypothesis that the reduction in in vivo JH titer in larval T. ni is due, in part, to hydrolysis of the hormone by selective esterases. DNTFP appears to be competing with JH for the active site of JH esterase.     

1 alpha, 25-Dihydroxyvitamin D3 directly induces fusion of alveolar macrophages by a mechanism involving RNA and protein synthesis, but not DNA synthesis

The results of our present study indicate that 1 alpha, 25-dihydroxyvitamin D3[1 alpha, 25(OH)2D3] directly induces fusion of mouse alveolar macrophages without any participation of T-lymphocytes by a mechanism involving RNA and protein synthesis but not DNA synthesis. We have reported that 1 alpha, 25(OH)2D3 induces fusion of alveolar macrophages by a direct mechanism and by a spleen cell-mediated indirect mechanism [(1983) Proc. Natl. Acad. Sci. USA 80, 5583-5587]. Alveolar macrophages pretreated with or without anti-Thy 1.2 antibody and complement fused similarly when they were incubated with 1 alpha, 25(OH)2D3. The vitamin suppressed DNA synthesis, but it significantly enhanced RNA and protein synthesis. The 1 alpha, 25(OH)2D3-induced fusion was blocked by adding actinomycin D or cycloheximide, but not by hydroxyurea.     

beta-Endorphin: structure-activity relationships in the guinea pig ileum and opiate receptor binding assays.

The opiate activities of some derivatives and enzymatic digests of camel and human β-endorphin were determined in the guinea pig ileum and rat brain opiate receptor binding assays. Derivatives of β-endorphins altered within the amino-terminal five residues showed pronounced losses in activity. Anisylation of the C-terminal glutamic acid residue of β_h-endorphin produced only small reductions in activity. Chymotryptic digestion greatly weakened the opiate activities of β_h-endorphin, whereas carboxypeptidase A, tryptic and leucine aminopeptidase digests showed only small losses in potency. The C-terminus of β-endorphin appears to contribute little directly to opiate activity. Amino acid analysis and assay of the leucine aminopeptidase digests suggest that the larger potency of β-endorphin relative to Met-enkephalin may be a consequence of its greater resistance to exopeptidase attack.     

Effect of inoculation withGlomus mosseae on nitrogen fixation by field grown soybeans

Summary This field study was undertaken to determine the effect of inoculation withGlomus mosseae on N₂ fixation and P uptake by soybean. The inoculation withGlomus mosseae was achieved using a new type of inoculant, alginate-entrapped (AE) endomycorrhizal fungus. N₂ fixation was assessed using the A value method. In P-fertilized plots, inoculation with AEGlomus mosseae increased the harvest index based on dry weight (+20%) and N content of seeds (+17%), the A value (+31%) and %N derived from fixation (+75%). Inoculation with AEGlomus mosseae decreased the coefficient of variation for the A value and for the dry weights of the different plant parts.     

Effects of microtubule-disruptive and membrane-stabilizing agents on low density lipoprotein metabolism by cultured human fibroblasts

The cellular mechanisms involved in the uptake and metabolism of low density lipoprotein (LDL) by cultured normal human fibroblasts have been investigated with the aid of drugs known to disrupt cytoplasmic microtubules or to inhibit membrane fusion.Two drugs which disrupt microtubules by differing mechanisms, colchicine and vinblastine, each reduced the high affinity surface binding of ¹²⁵I-labelled LDL by fibroblasts. Associated reductions of the endocytosis and degradation of the lipoprotein could be attributed almost entirely to this effect. In contrast, lumicolchicine, an analogue of colchicine without microtubule-disruptive activity, had little or no effect on ¹²⁵I-labelled LDL metabolism.Each of two groups of membrane-stabilizing agents, the phenothiazines and the tertiary amine local anaesthetics, directly inhibited both the internalization of ¹²⁵I-labelled LDL following high affinity binding to cell surface receptors and the catabolism of the lipoprotein subsequent to endocytosis, supporting previous morphological evidence for the importance of membrane fusion in these processes.