The effect ofMetarhizium anisopliae (DAT F-001) concentration and exposure time on the survival of the subterranean pasture pest,Adoryphorus couloni (Col.: Scarabaeidae)

The virulence of the DAT F-001 isolate ofMetarhizium anisopliae forAdoryphorus couloni was tested by exposing final instar larvae (L3) to concentrations of 10¹ to 10⁷ spores/g in a sand-peat mix for up to 112 days at 20°±2°C. All concentrations were pathogenic to L3 larvae and survival was dependent on concentration and correlated with exposure time. The time to kill 50% of the treated larvae (LT₅₀) ranged from 18.9 days (10⁷ spores/g) to 82.7 days (10¹ spores/g). The effect of DAT F-001 on feeding by L3 larvae was determined by comparing the production and weight of frass pellets/larva/day of untreated control larvae with larvae rolled in sporulating cultures ofM. anisopliae DAT F-001 and DAT F-054 (low virulence). Exposure to, and infection byM. anisopliae DAT F-001 had minimal impact on larval feeding. L3 larvae continued to feed at the same rate as both untreated and DAT F-054 treated larvae virtually until they died.

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Résumé La virulence deM. anisopliae DAT F-001 pourA. couloni a été analysée en exposant des larves L3 à des concentrations de 10¹ à 10⁷ spores/g dans un mélange de sable et de tourbe pendant un maximum de 112 jours à une température de 20°±2°C. Toutes les concentrations sont pathogènes pour les larves L3; la survie dépendait de la concentration et est en corrélation avec le temps d'exposition. Les LT₅₀ sont compris entre 18,9 jours (10⁷ spores/g) et 82,7 jours (10¹ spores/g). L'effet de DAT F-001 sur l'alimentation des larves L3 a été évalué en comparant la production et le poids des fèces/larve/jour des larves non-traitées avec des larves roulées dans des cultures sporulées deM. anisopliae DAT F-001 et DAT F-054 (faible virulence). L'exposition àM. anisopliae DAT F-001 suivie de l'infection a peu d'effet sur l'alimentation des larves. Les larves L3 continuent à s'alimenter au même rythme que les larves non-traitées de DAT F-054, quasiment jusqu'à leur mort.

     

Senescence mechanisms

Senescence in plants is usually viewed as an internally programmed degeneration leading to death. It is a developmental process that occurs in many different tissues and serves different purposes. Generally, apoptosis refers to programmed death of small numbers of animal cells, and it shows some special features at the cell level. Some senescing plant cells show some symptoms typical of apoptosis, while others do not. This review will focus primarily on leaf senescence with ultimate aim of explaining whole plant senescence (i.e., monocarpic senescence). Traditionally, the ideas on senescence mechanisms fall into two major groupings, nutrient deficiencies (e.g., starvation) and genetic programming (i.e., senescence-promoting and senescence-inhibiting genes). Considerable evidence indicates that nutrient deficiencies are not central senescence program components, while increasing evidence supports genetic programming. Because chlorophyll (Chl) and chloroplast (CP) breakdown are so prominent, leaf senescence is generally measured in terms of Chl loss. Although CP breakdown may not be the proximate cause of leaf cell death, it certainly is important as a source of nutrients for use elsewhere, e.g., for developing reproductive structures in monocarpic plants, and this loss limits assimilatory capacity. The CP is dismantled in an orderly sequence. Individual protein complexes seem to be taken out all at once, not one subunit at a time. Removal of any component, e.g., Chl, seems to destabilize the whole complex. It is of special interest that senescing CPs secrete Chl-containing globules indicating that some CP components are broken down outside the CP. Senescence appears to be imposed on the CP by the nucleus, and all the known senescence-altering genes except one, cytG in soybean, are nuclear. Only the d₁d₂ mutation(s) in soybean prevents a broad range of leaf senescence processes. Exactly, what causes cell death is unclear; however, the selective thiol protease inhibitor, E-64, does delay death, and this suggests that proteases play a key role.     

Petroleum bioremediation — a multiphase problem

Microbial degradation of hydrocarbons is a multiphase reaction, involving oxygen gas, water-insoluble hydrocarbons, water, dissolved salts and microorganisms. The fact that the first step in hydrocarbon catabolism involves a membrane-bound oxygenase makes it essential for microorganisms to come into direct contact with the hydrocarbon substrate. Growth then proceeds on the hydrocarbon/water interface. Bacteria have developed two general strategies for enhancing contact with water-insoluble hydrocarbons: specific adhesion mechanisms and production of extracellular emulsifying agents. Since petroleum is a complex mixture of many different classes of hydrocarbons, of which any particular microorganism has the potential to degrade only part, it follows that the microorganisms must also have a mechanism for desorbing from used' oil droplets.The major limitations in bioremediation of hydrocarbon-contaminated water and soil is available sources of nitrogen and phosphorus. The usual sources of these materials, e.g. ammonium sulfate and phosphate salts, have a high water solubility which reduces their effectiveness in open systems because of rapid dilution. We have attempted to overcome this problem by the use of a new controlled-release, hydrophobic fertilizer, F-1, which is a modified urea-formaldehyde polymer containing 18% N and 10% P as P₂O₅. Microorganisms were obtained by enrichment culture that could grow on crude oil as the carbon and energy source and F-1 as the nitrogen and phosphorus source. The microorganisms and the F-1 adhered to the oil/water interface, as observed microscopically and by the fact that degradation proceeded even when the water phase was removed and replaced seven times with unsupplemented water — a simulated open system. Strains which can use F-1 contain a cell-bound, inducible enzyme which depolymerizes F-1.After optimizing conditions in the laboratory for the use of F-1 and the selected bacteria for degrading crude oil, a field trial was performed on an oil contaminated sandy beach between Haifa and Acre, Israel, in the summer of 1992. The sand was treated with 5 g F-1 per kg sand and inoculated with the selected bacteria; the plot was watered with sea water and plowed daily. After 28 days the average hydrocarbon content of the sand decreased from 5.1 mg per g sand to 0.6 mg per g sand. Overall, there was an approx. 86% degradation of pentane extractables as demonstrated by dry weight, I.R. and GLC analyses. An untreated control plot showed only a 15% decrease in hydrocarbons. During the winter of 1992, the entire beach (approx. 200 tons of crude oil) was cleaned using the F-1 bacteria technology. The rate of degradation was 0.06 mg g^-1 sand day^-1 (10°C) compared to 0.13 mg g^-1 sand day^-1 during the summer (25°C).     

Investigations on feasibility of in situ development of amphotericin B liposomes for industrial applications

Amphotericin B (AmB) liposome formulations are very successful in the treatment of fungal infections and leishmaniasis. But higher cost limits its widespread use among people in developing countries. Therefore, we have developed a modified ethanol-injection method for the preparation of AmB liposomes. Two liposomal formulations were developed with dimyristoyl phosphatidylcholine [F-1a] and soya phosphatidylcholine [F-2a], along with egg phosphatidyl glycerol and cholesterol. AmB was dissolved in acidified dimethyl acetamide and mixed with ethanolic lipid solution and rapidly injected in 5% dextrose to prepare liposomes. Liposomes were characterized on the basis of size (~100?nm), zeta (–43.3?±?2.8 mV) and percent entrapment efficiency (>95%). The in vitro release study showed an insignificant difference (P?≥?0.05) for 24-hour release between marketed AmB liposomes (AmBisome) and F-1a and F-2a. Proliposome concentrate, used for the preparation of in situ liposomes, was physically stable for more than 3 months at experimental conditions. Similarly, AmB showed no sign of degradation in reconstituted liposomes stored at 2–8°C for more than 3 months. IC₅₀ value of Ambisome (0.18 µg/mL) was comparatively similar to F-1a (0.17 µg/mL) and F-2a (0.16 µg/mL) against intramacrophagic amastigotes. Under experimental conditions, a novel modified method for AmB liposomes is a great success and generates interest for development as a platform technology for many therapeutic drug products.     

DegePrime,a Program for Degenerate Primer Design for Broad-Taxonomic-Range PCR in Microbial Ecology Studies

The taxonomic composition of a microbial community can be deduced by analyzing its rRNA gene content by, e.g., high-throughput DNA sequencing or DNA chips. Such methods typically are based on PCR amplification of rRNA gene sequences using broad-taxonomic-range PCR primers. In these analyses, the use of optimal primers is crucial for achieving an unbiased representation of community composition. Here, we present the computer program DegePrime that, for each position of a multiple sequence alignment, finds a degenerate oligomer of as high coverage as possible and outputs its coverage among taxonomic divisions. We show that our novel heuristic, which we call weighted randomized combination, performs better than previously described algorithms for solving the maximum coverage degenerate primer design problem. We previously used DegePrime to design a broad-taxonomic-range primer pair that targets the bacterial V3-V4 region (341F-805R) (D. P. Herlemann, M. Labrenz, K. Jurgens, S. Bertilsson, J. J. Waniek, and A. F. Andersson, ISME J. 5:1571–1579, 2011, http://dx.doi.org/10.1038/ismej.2011.41), and here we use the program to significantly increase the coverage of a primer pair (515F-806R) widely used for Illumina-based surveys of bacterial and archaeal diversity. By comparison with shotgun metagenomics, we show that the primers give an accurate representation of microbial diversity in natural samples.     

Different cytotoxic and clastogenic effects of epigallocatechin gallate in various cell-culture media due to variable rates of its oxidation in the culture medium

Positive genotoxicity results are often observed using mammalian cells in culture with agents that are not in vivo genotoxins. We here illustrate one possible explanation: interaction of test chemicals with the cell-culture media used. We find that the toxicity and clastogenicity of epigallocatechin gallate (EGCG) to Chinese Hamster ovary (CHO) cells is affected by the culture medium used and appears largely or entirely due to variable rates of formation of hydrogen peroxide (H₂O₂) by chemical reactions of EGCG with the culture media. Catalase decreased EGCG toxicity substantially. Of seven different types of commonly used media evaluated, F-10 and F-12 nutrient mixtures were the least prone to produce this artefact. Although it generated H₂O₂ in the culture media, ascorbate was not toxic to CHO cells because the H₂O₂ levels achieved were insufficient to kill these cells. Thus, the culture medium, the cell type and the presence or absence of catalase (e.g. its variable amounts in S9 fractions) must be taken into account in in vitro genotoxicity testing.     

The Application of the Acoustic Complexity Indices (ACI) to Ecoacoustic Event Detection and Identification (EEDI) Modeling

In programs of acoustic survey, the amount of data collected and the lack of automatic routines for their classification and interpretation can represent a serious obstacle to achieving quick results. To overcome these obstacles, we are proposing an ecosemiotic model of data mining, ecoacoustic event detection and identification (EEDI), that uses a combination of the acoustic complexity indices (ACIt_f, ACIf_t, and ACIf_te) and automatically extracts the ecoacoustic events of interest from the sound files. These events may be indicators of environmental functioning at the scale of individual vocal species (e.g., behavior, phenology, and dynamics), the acoustic community (e.g., dawn and dusk chorus), the sound marks (e.g., the flag species of a community), or the soundscape (e.g., sonotope types). The EEDI model is represented by three procedural steps: 1) selecting acoustic data according to environmental variables, 2) detecting the events by creating an ecoacoustic event space (EES) produced by plotting ACIf_t and its evenness (ACIf_te), 3) identifying events according to the level of correlation between the acoustic signature (ACIt_f) of the detected events and an ad hoc library of previously identified events. The EEDI procedure can be extensively used in basic and applied research. In particular, EEDI may be used in long-term monitoring programs to assess the effect of climate change on individual vocal species behavior (fishes, frogs, birds, mammals, and arthropods), population, and acoustic community dynamics. The EEDI model can be also used to investigate acoustic human intrusion in natural systems and the effect in urban areas.     

The role of mast cell-derived secreted phospholipases A2 in respiratory allergy

Secreted phospholipases A₂ (sPLA₂s) are molecules released in plasma and biological fluids of patients with systemic inflammatory, autoimmune and allergic diseases. These molecules exert proinflammatory effects by either enzymatic-mechanisms or through binding to surface molecules expressed on inflammatory cells. sPLA₂s are released at low levels in the normal airways and tend to increase during respiratory allergies (e.g., rhinitis and bronchial asthma) as the result of local secretion. Several sPLA₂ isoforms are expressed in the human lung and some of them (e.g., group IIA and group X) are released in the airways of patients with rhinitis or asthma. Mast cells play a major role in the pathogenesis of respiratory allergies and other chronic inflammatory lung diseases. Recent evidence indicates that mast cells purified from human lung express most of the sPLA₂ isoforms so far described. IgE-mediated activation of these cells induce the release of sPLA₂s suggesting that mast cells are a main source of extracellular sPLA₂s during allergic reactions. Once released, sPLA₂s may contribute to the generation of eicosanoids (e.g., PGD₂ and LTC₄) and to the release of preformed mediators (e.g., histamine) by an autocrine loop involving the interaction of sPLA₂s with surface molecules such as heparan sulphate proteoglycans or the M-type receptor. Thus, mast cell-derived sPLA₂s may play an important role in the initiation and amplification of the inflammatory reactions in patients with allergic rhinitis and bronchial asthma.     

Effects of red blood cell aggregation on myocardial hematocrit gradient using two approaches to increase aggregation

The normal transmyocardial tissue hematocrit distribution (i.e., subepicardial greater than subendocardial) is known to be affected by red blood cell (RBC) aggregation. Prior studies employing the use of infused large macromolecules to increase erythrocyte aggregation are complicated by both increased plasma viscosity and dilution of plasma. Using a new technique to specifically alter the aggregation behavior by covalent attachment of Pluronic F-98 to the surface of the RBC, we have determined the effects of only enhanced aggregation (i.e., Pluronic F-98-coated RBCs) versus enhanced aggregation with increased plasma viscosity (i.e., an addition of 500 kDa dextran) on myocardial tissue hematocrit in rapidly frozen guinea pig hearts. Although both approaches equally increased aggregation, tissue hematocrit profiles differed markedly: 1) when Pluronic F-98-coated cells were used, the normal transmyocardial gradient was abolished, and 2) when dextran was added, the hematocrit remained at subepicardial levels for about one-half the thickness of the myocardium and then rapidly decreased to the control level in the subendocardial layer. Our results indicate that myocardial hematocrit profiles are sensitive to both RBC aggregation and to changes of plasma viscosity associated with increased RBC aggregation. Furthermore, they suggest the need for additional studies to explore the mechanisms affecting RBC distribution in three-dimensional vascular beds.     

Near‐Infrared,Heavy Metal‐Free Colloidal “Giant” Core/Shell Quantum Dots

“Giant” core/shell quantum dots (g‐QDs) are a promising class of materials for future optoelectronic technologies due to their superior chemical‐ and photostability compared to bare QDs and core/thin shell QDs. However, inadequate light absorption in the visible and near‐infrared (NIR) region and frequent use of toxic heavy metals (e.g., Cd and Pb) are still major challenges for most g‐QDs (e.g., CdSe/CdS) synthesized to date. The synthesis of NIR, heavy metal‐free, Zn‐treated spherical CuInSe₂/CuInS₂ g‐QDs is reported using the sequential cation exchange method. These g‐QDs exhibit tunable NIR optical absorption and photoluminescence (PL) properties. Transient fluorescence spectroscopy shows prolonged lifetime with increasing shell thickness, indicating the formation of quasi type‐II band alignment, which is further confirmed by simulations. As a proof‐of‐concept, as‐synthesized g‐QDs are used to sensitize TiO₂ as a photoanode in a photoelectrochemical (PEC) cell, demonstrating an efficient and stable PEC system. These results pave the way toward synthesizing NIR heavy metal‐free g‐QDs, which are very promising components of future optoelectronic technologies.     

Immunopontentiating and antitumor activities of the purified polysaccharides from Phellodendron chinese SCHNEID

The polysaccharide fractions were isolated and purified from Phellodendron chinese SCHNEID, and antitumor activities were examined at dosages of 2, 5 and 10 mg/100 g. F-7 and F-8 showed the highest tumor inhibitory activities (inhibition ratio 96.4 and 98.2% in 2 mg/100 g), and in dose of 5 mg/100 g, the inhibitory ratios were 95.3 and 97.5%, respectively. Furthermore, 10 mg/100 g of intraperitoneal (i.p.) injection gave 97.3 and 98.7% of inhibition. In oral administration, the inhibitory activities were not markedly observed, indicating that the polysaccharides are directly acting to immune system. Also the polysaccharides increased the number of circulating blood leukocytes and total peritoneal exudate cells. Although implantation of tumor cells greatly decreased the productivity of antibody (antibody-mediated) and T lymphocyte reactivity (delayed-type) as 6.3 from 9.3 and 5.9 from 7.7, represented by the increase of footpad thickness, respectively. The polysaccharides elevated the reactivity of T lymphocyte in tumor-bearing mice, which were rapidly recovered by discontinuance of sample treatments. Especially, F-2, F-5, F-7 and F-8 remarkably recovered the decreased sensitivity. When the effects on thymidylate synthase (TS) and thymidine kinase (TK) activities were determined, TS activities in the F-2 and F-7-treated mice were markedly suppressed to 73.7% and 79.5% of that in the control (p < 0.01), while there was little difference in TK activity with a slight decrease in F-2 only. However, in i.p. injection, TS activities in the F-2, F-5, F-7 and F-8-treated mice were markedly suppressed to 83% to 85% of that in the control (p < 0.01). Furthermore, there were also significant differences in TK activities in F-2, F-5, F-7 and F-8-treated mice (p < 0.05). These results clearly indicated that the i.p. injection is much effective to suppress tumor growth than oral administration.     

A method to accurately quantitate intensities of 32P-DNA bands when multiple bands appear in a single lane of a gel is used to study dNTP insertion opposite a benzo[a]pyrene-dG adduct by Sulfolobus DNA polymerases Dpo4 and Dbh

Quantitating relative ³²P-band intensity in gels is desired, e.g., to study primer-extension kinetics of DNA polymerases (DNAPs). Following imaging, multiple ³²P-bands are often present in lanes. Though individual bands appear by eye to be simple and well-resolved, scanning reveals they are actually skewed-Gaussian in shape and neighboring bands are overlapping, which complicates quantitation, because slower migrating bands often have considerable contributions from the trailing edges of faster migrating bands. A method is described to accurately quantitate adjacent ³²P-bands, which relies on having a standard: a simple skewed-Gaussian curve from an analogous pure, single-component band (e.g., primer alone). This single-component scan/curve is superimposed on its corresponding band in an experimentally determined scan/curve containing multiple bands (e.g., generated in a primer-extension reaction); intensity exceeding the single-component scan/curve is attributed to other components (e.g., insertion products). Relative areas/intensities are determined via pixel analysis, from which relative molarity of components is computed. Common software is used. Commonly used alternative methods (e.g., drawing boxes around bands) are shown to be less accurate. Our method was used to study kinetics of dNTP primer-extension opposite a benzo[a]pyrene-N²-dG-adduct with four DNAPs, including Sulfolobus solfataricus Dpo4 and Sulfolobus acidocaldarius Dbh. V_max/K_m is similar for correct dCTP insertion with Dpo4 and Dbh. Compared to Dpo4, Dbh misinsertion is slower for dATP (∼20-fold), dGTP (∼110-fold) and dTTP (∼6-fold), due to decreases in V_max. These findings provide support that Dbh is in the same Y-Family DNAP class as eukaryotic DNAP κ and bacterial DNAP IV, which accurately bypass N²-dG adducts, as well as establish the scan-method described herein as an accurate method to quantitate relative intensity of overlapping bands in a single lane, whether generated from ³²P-signals or by other means (e.g., staining).     

Structural requirements for anion substrates of the methotrexate transport system in L1210 cells

A broad spectrum of structurally diverse anions reversibly inhibits the influx of methotrexate in L1210 cells. Several of the more effective anions and their respective inhibition constants (K_i values) were: 5-methyltetrahydrofolate (0.3 μm), bromosulfophthalein (2 μm), thiamine pyrophosphate (3 μm), 8-anilino-1-naphthalene sulfonate (7 μm), phthalate (20 μm), and AMP (50 μm). Moderate inhibition was observed with P_i (K_i = 400 μm) and other divalent inorganic anions, while small monovalent anions such as Cl^? (K_i = 30 mm) were the least effective. When these same anions were tested for an effect on methotrexate efflux, stimulation was observed with some anions, while others had no effect. Enhancement was produced by folate compounds and p-aminobenzoylglutamate, small monovalent (e.g., Cl^?, acetate, and lactate) and divalent (e.g., phosphate and succinate) anions, a few nucleotides (e.g., AMP), and thiamine pyrophosphate, while little or no effect was associated with trivalent anions (e.g., citrate), most nucleotides, and large organic anions (e.g., bromosulfophthalein, NAD, and NADP). Anions with the ability to promote methotrexate efflux in control cells lost this capacity upon exposure of the cells to an irreversible inhibitor of methotrexate influx. These results support the hypothesis that methotrexate transport proceeds via an anion-exchange mechanism and moreover provide evidence that anion substrates for this system can be identified by their ability to promote methotrexate efflux. Anions which appear most likely to participate in this exchange cycle in vivo are P_i and AMP.     

Effect of detergents on the thermal behavior of elastin‐like polypeptides

Elastin‐like polypeptide (ELP) fusions have been designed to allow large‐scale, nonchromatographic purification of many soluble proteins by using the inverse transition cycling (ITC) method; however, the sensitivity of the aqueous lower critical solubility phase transition temperature (T_t) of ELPs to the addition of cosolutes, including detergents, may be a potential hindrance in purification of proteins with surface hydrophobicity in such a manner. To identify detergents that are known to solubilize such proteins (e.g., membrane proteins) and that have little effect on the T_t of the ELP, we screened a number of detergents with respect to their effects on the T_t and secondary structures of a model ELP (denoted here as ELP180). We found that mild detergents (e.g., n‐dodecyl‐β‐D ‐maltoside, Triton‐X100, and 3‐[(3‐cholamidopropyl) dimethylamino]‐1‐propanesulfonate) do not alter the phase transition behavior or structure (as probed by circular dichroism) of ELP180. This result is in contrast to previous studies that showed a strong effect of other detergents (e.g., sodium dodecylsulfate) on the T_t of ELPs. Our results clearly indicate that mild detergents do not preclude ITC‐based separation of ELPs, and thus that ELP fusions may prove to be useful in the purification of detergent‐solubilized recombinant hydrophobic proteins, including membrane proteins, which are otherwise notoriously difficult to extract and purify by conventional separation methods (e.g., chromatography). © 2012 Wiley Periodicals, Inc.     

Effect of Pluronic F-68 on the mechanical properties of mammalian cells

The mechanical properties of TB/C3 hybridoma cells taken from a continuous culture were measured by micromanipulation. The culture conditions were constant except for the presence or absence of Pluronic F-68 in the medium. It was found that the mean bursting membrane tension and the mean elastic area compressibility modulus of the cells were significantly greater (60% and 120%, respectively) in a medium with 0.05% (w/u) Pluronic F-68 compared to that without Pluronic. Pluronic F-68 therefore affected the strength of the membranes when the cells were exposed to it for a long period of time, i.e., in culture. The short-term effect of Pluronic F-68 on cell strength was also tested by its addition at various levels up to 0.2% (w/v) immediately before the mechanical property measurements. The resulting cell strength depended on the Pluronic concentration, but a significant short-term effect could only be detected above a threshold of 0.1% (w/v). Previous reports on the effect of Pluronic F-68 on animal cell culture are evaluated in the light of these observations.     

Regulation and roles for alternative pathways of hexose metabolism in plants

Plant cells have two cytoplasmic pathways of glycolysis and gluconeogenesis for the reversible interconversion of fructose 6-phosphate (F-6-P) and fructose 1,6-bisphosphate (F-1,6-P₂). One pathway is described as a maintenance pathway that is catalyzed by a nucleotide triphosphate-dependent phosphofructokinase (EC 2.7.1.11; ATP-PFK) glycolytically and a F-1,6 bisphosphatase (EC 3.1.3.11) gluconeogenically. These are non-equilibrium reactions that are energy consuming. The second pathway, described as an adaptive pathway, is catalyzed by a readily reversible pyrophosphate-dependent phosphofructokinase (EC 2.7.1.90; PP-PFK) in an equilibrium reaction that conserves energy through the utilization and the synthesis of pyrophosphate. A constitutive regulator cycle is also present for the synthesis and hydrolysis of fructose 2,6-bisphosphate (F-2,6-P₂) via a 2-kinase and a 2-phosphatase, respectively. The pathway catalyzed by the ATP-PFK and F-1,6-bisphosphatase, the maintenance pathway, is fairly constant in maximum activity in various plant tissues and shows less regulation by F-2,6-P₂. Plants use F-2,6-P₂ initially to regulate the adaptive pathway at the reversible PP_i-PFK step. The adaptive pathway, catalyzed by PP_i-PFK, varies in maximum activity with a variety of phenomena such as plant development or changing biological and physical environments. Plants can change F-2,6-P₂ levels rapidly, in less than 1 min when subjected to rapid environmental change, or change levels slowly over periods of hours and days as tissues develop. Both types of change enable plants to cope with the environmental and developmental changes that occur during their lifetimes. The two pathways of sugar metabolism can be efficiently linked by the cycling of uridylates and pyrophosphate required for sucrose breakdown via a proposed sucrose synthase pathway. The breakdown of sucrose via the sucrose synthase pathway requires half the net energy of breakdown via the invertase pathway. Pyrophosphate occurs in plant tissues as a substrate pool for biosynthetic reactions such as the PP_i-PFK or uridine diphosphate glucose pyrophosphorylase (EC 2.7.7.9; UDPG pyrophosphorylase) that function in the breakdown of imported sucrose. Also, pyrophosphate links the two glycolytic/gluco-neogenic pathways; and in a reciprocal manner pyrophosphate is produced as an energy source during gluconeogenic carbon flow from F-1,6-P₂ toward sucrose synthesis.     

Diminution of phagocytosed perfluorocarbon emulsions using perfluoroalkylated polyethylene glycol surfactant

Perfluorocarbon emulsions have been considered as potential blood substitutes for years due to their high capacity of dissolving respiratory oxygen and carbon dioxide. However, they have been reported to associate with side effects (e.g., flu-like syndrome) after being injected into animal's bloodstream. The cause of these side effects is related to the phagocytosis of perfluorocarbon emulsions by cells (e.g., macrophages). Inspired by the approach of using polyethylene glycol (PEG) to camouflage liposomes, we synthesized a perfluoroalkylated PEG (R(F)-PEG) surfactant to provide steric hindrance for decreasing phagocytosis of perfluorocarbon emulsions. The R(F)-PEG surfactant along with Pluronic F-68 and egg yolk phospholipid mediated perfluorocarbon emulsions were incubated individually with J774A.1 macrophages to examine the degree of phagocytosis. 19F NMR studies were used to quantitatively determine the amount of perfluorocarbon emulsions phagocytosed by macrophages. Results showed that the degree of phagocytosis was diminished to a large extent for perfluorocarbon microparticles emulsified by the R(F)-PEG surfactant.     

Strong Patterns in Homooligomer Tracts Occurrences in Non-Coding and in Potential Regulatory Sites in Eukaryotic Genomes

Abstract

Previous studies of the dinucleotides flanking both the 5′ and 3′ ends of homooligomer tracts have shown that some flanks are consistently preferred over others (1,2). In the first preferred group, the homooligomer tracts are flanked by the same nucleotide and/or the complementary nucleotides, e.g., ATA_n, TTA_n, CCG_n, where n=2–5. Runs flanked by nucleotides with which they cannot base pair are distinctly disfavored. (In this group A/T_n are flanked by C and/or G; G_n/C_n are flanked by A/T, e.g., CGA_n, T_nGG, G., AT). The frequencies of runs flanked by AorT, and G or C (“mixed” group) are as expected. Here we seek the origin of this effect and its relevance to protein-DNA interactions. Surprisingly, within the first group, runs flanked by their complements with a pyrimidine-purine junction (e.g., TTA_n, C_nGG) are greatly preferred. The frequencies of their purine-pyrimidine junction mirror-images is just as expected. This effect, as well as additional ones enumerated below, is seen universally in eukaryotes and in prokaryotes, although it is stronger in the former. Detailed analysis of regulatory regions shows these strong trends, particularly in GC sequences. The potential relationship to DNA conformation and DNA-protein interaction is discussed.     

Valuing the Benefits of Weight Loss Programs: An Application of the Discrete Choice Experiment

Objective: Obesity is a leading health threat. Determination of optimal therapies for long‐term weight loss remains a challenge. Evidence suggests that successful weight loss depends on the compliance of weight loss program participants with their weight loss efforts. Despite this, little is known regarding the attributes influencing such compliance. The purpose of this study was to assess, using a discrete choice experiment (DCE), the relative importance of weight loss program attributes to its participants and to express these preferences in terms of their willingness to pay for them. Research Methods: A DCE survey explored the following weight loss program attributes in a sample of 165 overweight adults enrolled in community weight loss programs: cost, travel time required to attend, extent of physician involvement (e.g., none, monthly, every 2 weeks), components (e.g., diet, exercise, behavior change) emphasized, and focus (e.g., group, individual). The rate at which participants were willing to trade among attributes and the willingness to pay for different configurations of combined attributes were estimated using regression modeling. Results: All attributes investigated appeared to be statistically significant. The most important unit change was “program components emphasized” (e.g., moving from diet only to diet and exercise). Discussion: The majority of participants were willing to pay for weight loss programs that reflected their preferences. The DCE tool was useful in quantifying and understanding individual preferences in obesity management and provided information that could help to maximize the efficiency of existing weight loss programs or the design of new programs.