Siderophores mediate reduced and increased uptake of cadmium by Streptomyces tendae F4 and sunflower (Helianthus annuus), respectively.

AIMS: As a toxic metal, cadmium (Cd) affects microbial and plant metabolic processes, thereby potentially reducing the efficiency of microbe or plant-mediated remediation of Cd-polluted soil. The role of siderophores produced by Streptomyces tendae F4 in the uptake of Cd by bacteria and plant was investigated to gain insight into the influence of siderophores on Cd availability to micro-organisms and plants. METHODS AND RESULTS: The bacterium was cultured under siderophore-inducing conditions in the presence of Cd. The kinetics of siderophore production and identification of the siderophores and their metal-bound forms were performed using electrospray ionization mass spectrometry. Inductively coupled plasma spectroscopy was used to measure iron (Fe) and Cd contents in the bacterium and in sunflower plant grown in Cd-amended soil. Siderophores significantly reduced the Cd uptake by the bacterium, while supplying it with iron. Bacterial culture filtrates containing three hydroxamate siderophores secreted by S. tendae F4 significantly promoted plant growth and enhanced uptake of Cd and Fe by the plant, relative to the control. Furthermore, application of siderophores caused slightly more Cd, but similar Fe uptake, compared with EDTA. Bioinoculation with Streptomyces caused a dramatic increase in plant Fe content, but resulted only in slight increase in plant Cd content. CONCLUSION: It is concluded that siderophores can help reduce toxic metal uptake in bacteria, while simultaneously facilitating the uptake of such metals by plants. Also, EDTA is not superior to hydroxamate siderophores in terms of metal solubilization for plant uptake. SIGNIFICANCE AND IMPACT OF THE STUDY: The study showed that microbial processes could indirectly influence the availability and amount of toxic metals taken up from the rhizosphere of plants. Furthermore, although EDTA is used for chelator-enhanced phytoremediation, microbial siderophores would be ideal for this purpose.

A type III polyketide synthase from Wachendorfia thyrsiflora and its role in diarylheptanoid and phenylphenalenone biosynthesis.

Chalcone synthase (CHS) related type III plant polyketide synthases (PKSs) are likely to be involved in the biosynthesis of diarylheptanoids (e.g. curcumin and polycyclic phenylphenalenones), but no such activity has been reported. Root cultures from Wachendorfia thyrsiflora (Haemodoraceae) are a suitable source to search for such enzymes because they synthesize large amounts of phenylphenalenones, but no other products that are known to require CHSs or related enzymes (e.g. flavonoids or stilbenes). A homology-based RT-PCR strategy led to the identification of cDNAs for a type III PKS sharing only approximately 60% identity with typical CHSs. It was named WtPKS1 (W. thyrsiflora polyketide synthase 1). The purified recombinant protein accepted a large variety of aromatic and aliphatic starter CoA esters, including phenylpropionyl- and side-chain unsaturated phenylpropanoid-CoAs. The simplest model for the initial reaction in diarylheptanoid biosynthesis predicts a phenylpropanoid-CoA as starter and a single condensation reaction to a diketide. Benzalacetones, the expected release products, were observed only with unsaturated phenylpropanoid-CoAs, and the best results were obtained with 4-coumaroyl-CoA (80% of the products). With all other substrates, WtPKS1 performed two condensation reactions and released pyrones. We propose that WtPKS1 catalyses the first step in diarylheptanoid biosynthesis and that the observed pyrones are derailment products in the absence of downstream processing proteins.

Sex pheromone of horse-chestnut leafminer Camneraria ohridella and its use in a pheromone-based monitoring system.

Gas chromatography combined with electroantennographic detection (GC-EAD), electroantennography (EAG), and wind-tunnel and field experiments were used to reinvestigate the composition of Cameraria ohridella (Lepidoptera, Gracillariidae, Lithocolletinae) sex pheromone. The GC-EAD experiments showed one EAD-active area corresponding to the major pheromone component. (8E,10Z)-tetradeca-8,10-dienal. The EAG experiments proved that (9E)-tetracedecenal and stereoisomers of (8E,10Z)-tetradeca-8,10-dienal exhibited significant electrophysiological activity and could, therefore, be considered as possible minor pheromone components. However, wind-tunnel and field experiments demonstrated that none of these compounds affect the efficacy of the main pheromone component. A monitoring system based on (8E,10Z)tetradeca-8,10-dienal was developed and used to study the flight activity of C. ohridella.

Evaluation of the synthetic major component of the sex pheromone of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae).

In wind-tunnel bioassays, dispensers loaded with 1 microg of the synthetic major component (3E,8Z, 11Z)-3,8,11-tetradecatrienyl acetate (TDTA) of the sex pheromone emitted by Tuta absoluta (Meyrick) females were found to be highly attractive to conspecific males. Field experiments were conducted to evaluate the efficacy of five trap designs. The best trap, baited with 100 microg of the synthetic sex pheromone caught on average 1,200 males per trap per night, while those baited with virgin females caught only 201 males. The male response to this pheromone is restricted to the same early-morning time window during which females exhibit calling behavior. The high biological activity of the synthetic pheromone suggests that it could be useful for pest monitoring and in mating disruption.

Detection and removal of an artefact fatty acid from the binding site of recombinant Bombyx mori pheromone-binding protein.

Recombinant Bombyx mori pheromone-binding protein (PBP), purified from an Escherichia coli expression system, has been found to contain (11Z)-octadecenoic acid (cis-vaccenic acid) as an artefact ligand. An efficient delipidation procedure is described to overcome what would appear to be a general problem with recombinant lepidopteran PBPs.

The divergence in the conception of Pavlov and Bayliss-Starling concerning the function of the nervous system.

The history of the discoveries of the mechanisms of the pancreatic secretion is a fine example of the conflict of two schools and the concurrence of two concepts: The nerve control (Pavlov and his co-workers) and the humoral control (Bayliss and Starling). Only the synthesis of both conceptions brought about the most accurate model of the control mechanisms.

Characterization of vinyl-substituted, carbon-carbon double bonds by GC/FT-IR analysis.

Vapor-phase infrared spectra allow the determination of the stereochemistry of carbon-carbon double bonds conjugated with a vinyl group. Cis and trans isomers of unsubstituted 1,3-alkadienes can be differentiated on the basis of the differences observed in the 900-1000 cm-1 region (spectra of cis isomers show two bands at 993 and 906 cm-1, while those of trans compounds show three absorptions at 998, 949, and 902 cm-1) and the 1590-1650 cm-1 region (the C=C stretch bands are observed at 1595 and 1642 cm-1 for cis compounds and at 1604 and 1650 cm-1 for trans compounds). Compounds bearing CH2=CHC(CH3)=CHCH2- and CH2=CHC(=CH2)-CH2- structural moieties, referred to as alpha- and beta-type compounds, are frequently encountered as natural products. For compounds bearing alpha-type groups, the cis/trans configuration of the trisubstituted double bond can be determined unambiguously. An absorption at 3095-3091 cm-1, for the =CH2 stretch vibration, is common to both of these groups; however, due to the presence of two =CH2 groups, the relative intensity of the band is much higher for beta-type compounds. For alpha-type compounds, a cis configuration at the C-3 carbon atom is characterized by a =CH2 wag absorption at 907-906 cm-1. For beta-type compounds and 3E-alpha-type compounds, this band appears at 899-897 cm-1. In addition, a wavy "fingerprint" pattern with two minima at 1632 (low intensity) and 1595-1594 cm-1 (high intensity) is characteristic for beta-type compounds. Our generalizations are based on spectra of cis and trans ocimene, myrcene, and dehydration products of many 3-methyl-1-alken-3-ols. Six isomers of farnesene can be characterized by GC/FT-IR. Furthermore, gas-phase IR allows the determination of the configuration of the trisubstituted double bond at C-3 in alpha-type farnesene congeners. For example, the homo- and bishomofarnesene isomers from Myrmica ants were shown to include a 3Z bond.

[Changes in elastase and collagenase serum levels in patients with sclerosing connective tissue diseases treated with uropancreozymin]. / Zmeny koncentrace elastázy a kolagenázy v krevním séru nemocyných s chorobami s tvorbou sklerotické vazivové tkáne behem lécby uropankreozyminem.

BACKGROUND: The objective of this work is to clarify the mechanism of the therapeutic effect of uropancreozymin upon the sclerosal process. METHOD AND RESULTS: The sick suffering from sclerosed connective tissue formation underwent uropancreozymin treatment. Their elastase and collagenase blood serum level was contemporaneously determined. One day after the administration of uropancreozymin the elastase level rose by 144.6%, after the third day by 187.4%, after the seventh day by 150.2%. The collagenase activity was elevated after the first day of treatment to 234.7%, after the third day by 187.4%, after the seventh day by 150.2%. The collagenase activity was elevated after the first day of treatment to 144.6%, after the third day to 132.7%, and after the seventh day to 150.8%. CONCLUSIONS: The therapeutic effect of uropancreozymin is explained as the faculty to wash out the pancreatic enzymes in active form, mainly elastase et collagenase, into the blood circulation. So it differs from pancreozymin which does not elevate the pancreatic enzyme level in blood in these diseases and which contains more hormonal factors. For this reason uropancreozymin is therapeutic and pancreozymin diagnostic. Pancreozymin was later mistakenly characterized as cholecystokinin (4.9) but this error has been already repaired (17).

(3E,8Z,11Z)-3,8,11-tetradecatrienyl acetate, major sex pheromone component of the tomato pest Scrobipalpuloides absoluta (Lepidoptera: Gelechiidae).

The major sex attractant emitted by Scrobipalpuloides absoluta females is shown to be (3E,8Z,11Z)-3,8,11-tetradecatrien-1-yl acetate by a novel strategy involving the random reduction of double bonds, followed by methylthiolation of the reduced products. Each female sex gland contains ca. 1-5 ng of this pheromone. This triene ester, synthesized by a stereospecific procedure, shows spectral and gas chromatographic properties identical to those of the natural substance. In field tests and wind tunnel bioassays, the synthetic ester was found to be highly attractive to conspecific males. The male response to this pheromone, however, is restricted to the same early-morning time window during which females exhibit calling behavior.

New mimics of the acetate function in pheromone-based attraction.

Several analogues of (Z)-8-dodecenyl acetate (1a), the major pheromone component of the Oriental fruit moth, Cydia molesta, with chloroformate and lactone functional groups in place of the acetate moiety, were synthesized and investigated for their biological activity at four evaluation levels, i.e. by electroantennography (EAG), electrosensillography (ESG), short-range sexual stimulation and activation in the flight-tunnel. We found very strict requirements on the shape as well as on the electron distribution of the acetate group for a productive interaction with the receptor. The behavioral results showed that, among the analogues investigated, the chloroformate 1b, alken-4-olide 2a and also dodecyl acetate (1c) possess significant (60-85%) inhibitory activities. Based on electrophysiological evidence demonstrating that (i) only 1b is competing with the major pheromone component 1a for the same receptor sites on the male antennal sensilla, (ii) 1c elicits moderate EAG but no ESG responses and (iii) 2a does not produce any electrophysiological response at all, three possible inhibitory mechanisms by which these analogues are acting could be distinguished.

Sex pheromone of tomato pestScrobipalpuloides absoluta (Lepidoptera: Gelechiidae).

The sex attractant ofScrobipalpuloides absoluta females is a 90:10 mixture of (3E,8Z,11Z)-3,8,11-tetradecatrien-1-yl acetate and (3E,8Z)-3,8-tetradecadien-1-yl acetate. Tetradecadienyl acetates bearing 8Z,11Z; 3E,8Z; and 3E,11Z double bonds were synthesized by stereospecific procedures; the mass spectral and gas chromatographic properties of the 3E,8Z isomer were found to be congruent with those of the tetradecadienyl acetate fromS. absoluta. In wind tunnel bioassays, a 10:1 mixture of synthetic (3E,8Z,11Z)-3,8,11-tetradecatrien-1-yl acetate and (3E,8Z)-3,8-tetradecadien-1-yl acetate was highly attractive toS. absoluta males. Interestingly, the presence of (8Z,11Z)-8,11-tetradecadien-1-yl acetate (10%) inhibited the response to (3E,8Z,11Z)-3,8,11-tetradecatrien-1-yl acetate completely.

Gas-phase infrared spectroscopy for determination of double bond configuration of monounsaturated compounds.

Gas-phase Fourier-transform infrared spectra allow unambiguous determination of the configuration of the double bonds of long-chain unsaturated compounds bearing RCH=CHR' type bonds. Although the infrared absorption at 970-967 cm-1 has been used previously for the identification of trans bonds, the absorption at 3028-3011 cm-1 is conventionally considered to be incapable of distinguishing cis and trans isomers. In this paper, we present a large number of gas-phase spectra of monounsaturated long-chain acetates which demonstrate that an absorption, highly characteristic for the cis configuration, occurs at 3013-3011 cm-1, while trans compounds fail to show any bands in this region. However, if a double bond is present at the C-2 or C-3 carbon atoms, this cis=CH stretch absorption shows a hypsochromic shift to 3029-3028 and 3018-3017 cm-1, respectively. Similarly, if a cis double bond is present at the penultimate carbon atom, this band appears at 3022-3021 cm-1. All the spectra of trans alkenyl acetates showed the expected C-H wag absorption at 968-964 cm-1. In addition, the spectra of (E)-2-alkenyl acetates show a unique three-peak "finger-print" pattern which allows the identification of the position and configuration of this bond. Furthermore, by synthesizing and obtaining spectra of appropriate deuteriated compounds, we have proved that the 3013-3011 cm-1 band is representative of the C-H stretching vibration of cis compounds of RCH=CHR' type.

[The effect of amino acids, sugars and fats on activation of pancreatic enzymes]. / Vliv aminokyselin, cukru a tuku na aktivaci pankreatických enzymu.

BACKGROUND: Investigations of pancreatic enzymes have again become a topic. Attention concentrates rather on exogenous activators. The objective of the present work was to assess to what extent some amino acids, sugars and fats affect the activity of pancreatic enzymes "in vitro". METHODS AND RESULTS: The author compared the degree of hydrolysis of enzyme-substrate with the degree of hydrolysis enzyme-substrate-activator. Twenty-four amino acids, 25 sugars and 7 fats were analyzed. Of the 24 tested amino acids only six activate amylase: cysteine 39.9 x, histidine 21 x, lysine 3.8 x, methionine 3.3 x, tryptophan 2.9 x and tyrosine 1.5 x. As compared with control groups, the differences are significant, p < 0.01 with the exception of tyrosine (p > 0.05). All amino acids activate lipase: serine 3.4 x, aspartic acid 3.1 x, asparagine 3.0 x, hydroxyproline 2.7 x alanine 1.1 x (p > 0.05). Of the investigated sugars none activated amylase, 14 do not activate picase. Fructose activates lipase 1.9 x, inosine 1.8 x, laevulose 1.7 x, maltose 1.6 x, sucrose 1.3 x, xylitol 1.2 x, sorbitol 1.1 x, glucosamine 2.6 x and galactosamine 2.0 x. Pure fats do not activate pancreatic enzymes. Butter causes a rise of trypsin activity by 1.3 x, lard of lipase activity 2 x and trypsin 6.2 x. CONCLUSIONS: In in vitro experiments evidence was provided that cysteine, histidine lysine and methionine tryptophan and tyrosine activate amylase, all 24 tested amino acids activate lipase as well as trypsin to a varying extent. The investigated sugars do not activate amylase at all; lipase is activated by 10 sugars and trypsin by four sugars. Pure fats do not activate any pancreatic enzymes.

[Sephadex G 100- Separation of the mucicrinic and the mucimitiginic factors from an extract of duodenal mucosa (author's transl)]. / Séparation sur séphadex G 100 de facteurs mucicriniques et mucimitiginiques à partir d'un extrait de muqueuse duodénale

The mucicrin stimulating the hexosamine gastrointestinal secretion and extracted form porcine duodenum, was concentrated in the zone 7 using sephadex G 100. The molecular weight of 4000-5000 corresponds to this zone. An opposite activity was detected in zone 3. The factor from digestive mucous origin being able to diminish the hexosamine gastric secretion and not influencing the arterial pressure was called mucimitigin. The molecular weight of about 35000 corresponds to this factor.