Form and environment of Gryphaea arcuata

Gryphaea arcuata is one of the most studied fossils, but its detailed palaeoecology has been largely neglected. Specimens were collected within a short stratigraphic range (three ammonite zones) in the 'Calcaire à gryphées' of Xeuilley (Lorraine, France) dated Hettangian to Lower Sinemurian. As far as possible, they were sampled from each marly bed of the section. A biometric study and an isotopic analysis are compared in regard to organic matter measurements and palynological data, the results demonstrating a clear relationship between the shape of G. arcuata and environmental parameters. Factors responsible for the various shapes are temperature, oxygen levels on the sea floor and nutrient levels. Two main morphotypes can be related to two kinds of environment. In the first, controlled by a relatively hot and humid climate and tending towards eutrophication, the growth rate of Gryphaea was low, and the shells small, wide and thin. In the second environment, cooler than the first one and closer to the optimal living conditions of G. arcuata, the shell was large, thick and narrow, and exhibited a high growth rate.     

Rates and equilibria at the acetylcholine receptor of electrophorus electroplaques. A study of neurally evoked postsynaptic currents and of voltage-jump relaxations

Kinetic measurements are employed to reconstruct the steady-state activation of acetylcholine [Ach] receptor channels in electrophorus electroplaques. Neurally evoked postsynaptic currents (PSCs) decay exponentially; at 15 degrees C the rate constant, α, equals 1.2 ms(-1) at 0 mV and decreases e-fold for every 86 mV as the membrane voltage is made more negative. Voltage-jump relaxations have been measured with bath-applied ACh, decamethonium, carbachol, or suberylcholine. We interpret the reciprocal relaxation time 1/τ as the sum of the rate constant α for channel closing and a first-order rate constant for channel opening. Where measureable, the opening rate increases linearly with [agonist] and does not vary with voltage. The voltage sensitivity of small steady-state conductances (e- fold for 86 mV) equals that of the closing rate α, confirming that the opening rate has little or no additional voltage sensitivity. Exposure to α-bungarotoxin irreversibly decreases the agonist-induced conductance but does not affect the relaxation kinetics. Tubocurarine reversibly reduces both the conductance and the opening rate. In the simultaneous presence of two agonist species, voltage-jump relaxations have at least two exponential components. The data are fit by a model in which (a) the channel opens as the receptor binds the second in a sequence of two agonist molecules, with a forward rate constant to 10(7) to 2x10(8) M(-1)s(-1); and (b) the channel then closes as either agonist molecule dissociates, with a voltage-dependent rate constant of 10(2) to 3x10(3)s(-1).     

Activities of Various Peptidases in the First Leaf of Wheat

In the first leaf of wheat (Triticum aestivum L. cv. Capelle) the content of soluble protein diminished to about 50% of the initial value between the 7th and the 19th day after sowing. In order to understand proteolysis in the leaves, the activities of several peptidases were measured in extracts from leaves at four different ages. The carboxypeptidase activities increased during the growth of the leaves, and then began to decrease. The activities of the alkaline peptidases increased, while that of the benzoyl-DL-arginine-p-nitroanilide (BAPA) hydrolysing enzyme decreased during the whole period studied. The “naphthylamidase” activities showed first a slow rise, but then leveled off. Two bands with naphthylamidase activity could be detected after disc electrophoresis. All the peptidases studied were present in the leaves at rather high concentrations. This indicates that they all may participate in the hydrolysis of leaf proteins into free amino acids.     

INSULIN PRODUCES A BIPHASIC RESPONSE INTETRAHYMENA THERMOPHILABY STIMULATING CELL SURVIVAL AND ACTIVATING PROLIFERATION IN TWO SEPARATE CONCENTRATION INTERVALS

Cells ofTetrahymenamay produce autocrine signal molecules with effects on survival and proliferation. Here we have tested the effects of human recombinant and bovine insulin, and the B22–B30 fragment of bovine insulin over a wide range of concentrations (10^?5–10^?18m ) on cell survival and proliferation in a synthetic nutrient medium. The cells were grown in conical flasks at low initial cell densities (40 and 400cells/ml). Insulin prevented rapid cell death and/or promoted cell proliferation over two separate concentration ranges: down to nanomolar levels and again in the low pico- and femtomolar range. At an initial population density of 400cells/ml the cells multiplied at both concentration intervals. At 40 or fewer organisms/ml the cells multiplied in the high concentration interval, whereas in the low interval they survived for about four times longer than those in the control cultures. B22–B30 added to cultures of 40 initial cells/ml produced a stimulation of cell survival in the low pico- and high femtomolar range. In the presence of hemin (50nm ) cells at 400 initial organisms/ml multiplied at insulin concentrations down to about 3nm and again from 300am to 10pm . In some cases, hemin plus insulin activated cell proliferation between the two concentration intervals as well. At 40cells/ml the cells not only survived but proliferated in the femtomolar range. Cells in cultures supplemented with both hemin and B22–B30 multiplied at the low concentration interval (from about 100fm to 10pm ).     

Fermentative hydrolysate of mycelial waste of aminoglycoside antibiotics production as component of culture media used in biosynthesis of tobramycin]

New nutrient media for cultivation of the tobramycin-producing organism were developed. As an additional source of nitrogen the media contain fermentative hydrolysate of the mycelial waste of manufacture of aminoglycoside antibiotics (tobramycin and apramycin). The use of the media provided a 20 to 50% decrease of consumption of soybean meal, an essential food raw material, and design of a low-waste technology for biosynthesis of tobramycin.     

PCR: how to kill unwanted DNA.

Avoidance of contamination in the PCR laboratory requires the use of strict precautions. Among these, chemical decontamination of surfaces and equipment is desirable to prevent inadvertent contamination of samples by the gloved hand and by pipettors. We have investigated the use of sodium hypochloride (Clorox), in comparison to concentrated HCl, for PCR sterilization. Ten percent Clorox was found to eliminate all ethidium bromide-stainable DNA and to prevent PCR amplification of a 600-bp DNA segment within one minute of template treatment. RNA was similarly destroyed. By contrast, even 2.0 N HCl did not destroy DNA detectable by PCR within five minutes. Because of its high efficacy, low cost and relatively low corrosiveness, we recommend the use of ten percent Clorox as a decontaminant for elimination of DNA templates in the PCR laboratory.     

Sixty simple sequence repeat markers for use in the Festuca–Lolium complex of grasses

So far only very few simple sequence repeat (SSR) markers developed from grass species have had their primer sequences published. To make more markers available to the scientific community, we isolated and sequenced 256 microsatellite‐containing clones from four genome libraries of a Lolium multiflorum×Festuca glaucescens F₁ hybrid following enrichment in (TC)_n, (TG)_n, or both repeats. In this work, we report the primer sequences of 60 SSRs including preliminary results of polymorphism for mapping.     

Base Modification and the Phosphoramidite Approach

Abstract

Oligodeoxynucleotides 18–150 bases in length were synthesized with both 0-(2-cyanoethyl)- and 0-(methyl)-phosphoramidites. After enzymatic degradation of the purified products, base modification and composition were evaluated by HPLC. Additionally, synthesis of 5′-d[GCGCGCTT] with O-(methyl) phosphorus protection generated 3-methylthymidine when thiophenoxide was omitted from the deprotection protocol.     

Purification of 5′-O-Trityl-on Oligoribonucleotides. Investigation of Phosphate Migration During Purification and Detritylation.

Abstract

Synthetic oligoribonucleotides (RNA) are efficiently prepared with 2′-O-tert-butyldimethylsilyl nucleoside 3′-O-phosphoramidites with labile base-protection; A_dmf or A_Pac, G_dmf, C_ibu, U. After cleavage from the polystyrene support, the exocyclic amine protecting groups are removed with conc. NH₄OH: ethanol/3:1 by heating at 55°C for 3–5 h. The 2′-O- silyl protecting groups are removed with tetra-n-butylammonium fluoride in THF or more conveniently with neat triethylamine trihydrofluoride. To gain the advantages of increased capacity on reverse phase HPLC and the convenience of cartridge based purification (OPC, Oligonucleotide Purification Cartridge), the 5′ trityl was left on the RNA as the final protecting group to be removed. The mild conditions which are effective for trityl removal are shown to preserve 3′-5′ phosphate linkage integrity in RNA. The absence of phosphate migration is demonstrated by model studies, utilizing N⁴ -isobutyryl-5′-O-DMT-3′-O-TBDMS-2′-O-(2-cyanoethyl-N,N-diisopropylphosphoramidite) as a control monomer and digestion by 3′-5′ selective P1 nuclease and alkaline phosphatase and HPLC analysis. Oligoribonucleotides were analyzed by Microgel capillary electrophoresis, anion-exchange HPLC, and the enzymatic digest/HPLC method.

     

悬钩子属植物化学成分及药理活性研究进展

本文对近10年来悬钩子属植物的化学成分和药理活性研究进行了综述,为该属植物的进一步开发利用提供参考。悬钩子属植物的化学成分主要包括黄酮、萜、鞣质、甾等。药理活性主要包括抗菌、抗炎、抗肿瘤、抗氧化、抗过敏、保肝、镇痛等。