Anti-Pseudomonas aeruginosa activity of hemlock (Conium maculatum, Apiaceae) essential oil.

Conium maculatum is a nitrophilous weed belonging to the Apiaceae family and occurring in hedgerows, pastures, waste ground, along rivers and roadsides. Little is known on the chemistry and bioactivity of other secondary metabolites occurring in the plant. In the present work, we have analysed the chemical composition and antimicrobial activity of the essential oils hydrodistilled from leaves and inflorescenes of C. maculatum growing in Sicily, Italy. The composition of essential oils was achieved by gas chromatography-mass spectrometry (GC-MS) analysis, whereas the inhibitory effects on the growth of two Gram negative strains, namely Escherichia coli and Pseudomonas aeruginosa were assessed by two different analysis. The essential oils exhibited different chemical profiles (1-butylpiperidine and myrcene in the inflorescenes), (mostly (E)-caryophyllene in the leaves). The latter oil was particularly active in inhibiting the growth of P. aeruginosa. These results shed light on the possible application of hemlock essential oils as antimicrobial agents.

Hemlock alkaloids from Socrates to poison aloes.

Hemlock (Conium maculatum L. Umbelliferae) has long been known as a poisonous plant. Toxicity is due to a group of piperidine alkaloids of which the representative members are coniine and gamma-coniceine. The latter is the more toxic and is the first formed biosynthetically. Its levels in relation to coniine vary widely according to environmental conditions and to provenance of the plants. Surprisingly, these piperidine alkaloids have turned up in quite unrelated species in the monocotyledons as well as the dicotyledons. Aloes, for instance, important medicinal plants, are not regarded as poisonous although some species are very bitter. Nevertheless a small number of mostly local species contain the alkaloids, especially gamma-coniceine and there have been records of human poisoning. The compounds are recognized by their characteristic mousy smell. Both acute and chronic symptoms have been described. The compounds are neurotoxins and death results from respiratory failure, recalling the effects of curare. Chronic non-lethal ingestion by pregnant livestock leads to foetal malformation. Both acute and chronic toxicity are seen with stock in damp meadows and have been recorded as problems especially in North America. The alkaloids derive biosynthetically from acetate units via the polyketide pathway in contrast to other piperidine alkaloids which derive from lysine.

Exploring the structural basis of neurotoxicity in C(17)-polyacetylenes isolated from water hemlock.

Water hemlock, Cicuta virosa, belonging to the Umbelliferae, is well-known as a toxic plant responsible for lethal poisonings in humans as well as animals, causing tonic and clonic convulsions and respiratory paralysis. Cicutoxin (1), being a major violent toxin of the plant, is a chemical in the class of C(17)-polyacetylenes bearing a long pi-bond conjugation system, a terminal hydroxyl, and an allylic hydroxyl in its structure, and a variety of its analogues have been isolated from the plant. In the present study, various derivatives of these toxins were synthesized through acetylation, methylation, and oxidation of cicutoxin (1) and virol A (3) and B (4). 1-Dehydroxyvirol A (28) was prepared through the coupling of (7S)-dodeca-3,5-dien-1-yn-7-ol and 1-iodopentyne under Sonogashira's conditions. A monoacetylenic compound (29) was also prepared through the coupling of (5S)-1-chlorodeca-1,3-dien-5-ol and 1-iodopentyn-5-ol. The structure-activity relationships involved in the acute toxicity of cicutoxin derivatives in mice were investigated, and the length and geometry of pi-bond conjugation and the O-functional groups were found to be important for activity. The potency in inhibition of the specific binding of the noncompetitive GABA antagonist, [(3)H]EBOB, to GABA-gated Cl(-) channels of GABA receptors in rat brain cortex was found to be correlated with acute toxicity, indicating that the ability to bind to these channels plays an important role in the acute toxicity of these compounds.

Biochemistry of hemlock (Conium maculatum L.) alkaloids and their acute and chronic toxicity in livestock. A review.

The literature on Conium maculatum biochemistry and toxicology, dispersed in a large number of scientific publications, has been put together in this review. C. maculatum is a weed known almost worldwide by its toxicity to many domestic animals and to human beings. It is an Umbelliferae, characterized by long, hollow stems, reaching up to 2 m height at maturity, producing a large amount of lush foliage during its vegetative growth. Its flowers are white, grouped in umbels formed by numerous umbellules. It produces a large number of seeds that allow the plant to form thick stands in modified soils, sometimes encroaching on cultivated fields, to the extent of impeding the growth of any other vegetation inside the C. maculatum area of growth. Eight piperidinic alkaloids have been identified in this species. Two of them, gamma-coniceine and coniine are generally the most abundant and they account for most of the plant acute and chronic toxicity. These alkaloids are synthesized by the plant from eight acetate units from the metabolic pool, forming a polyketoacid which cyclises through an aminotransferase and forms gamma-coniceine as the parent alkaloid via reduction by a NADPH-dependent reductase. The acute toxicity is observed when animals ingest C. maculatum vegetative and flowering plants and seeds. In a short time the alkaloids produce a neuromuscular blockage conducive to death when the respiratory muscles are affected. The chronic toxicity affects only pregnant animals. When they are poisoned by C. maculatum during the fetuses organ formation period, the offspring is born with malformations, mainly palatoschisis and multiple congenital contractures (MCC; frequently described as arthrogryposis). Acute toxicity, if not lethal, may resolve in the spontaneous recovery of the affected animals provided further exposure to C. maculatum is avoided. It has been observed that poisoned animals tend to return to feed on this plant. Chronic toxicity is irreversible and although MCC can be surgically corrected in some cases, most of the malformed animals are lost. Since no specific antidote is available, prevention is the only way to deal with the production loses caused by this weed. Control with herbicides and grazing with less susceptible animals (such as sheep) have been suggested. C. maculatum alkaloids can be transferred to milk and to fowl muscle tissue through which the former can reach the human food chain. The losses produced by C. maculatum chronic toxicity may be largely underestimated, at least in some regions, because of the difficulty in associate malformations in offspring with the much earlier maternal poisoning.

Effect of temperature and host tree on cold hardiness of hemlock looper eggs along a latitudinal gradient.

The hemlock looper, Lambdina fiscellaria, is an economically important insect pest of Canadian forests which overwinters as eggs. Although the hemlock looper causes extensive damages, no information on the mechanisms related to its cold tolerance is known. The objective of this study was to determine the effect of temperature and exposure duration on hemlock looper winter survival but also to identify seasonal supercooling capacity and cryoprotectant levels of three populations along a latitudinal gradient. As host plant may contribute to offspring overwintering success, cold tolerance of hemlock looper eggs from parents whose larvae were fed on three different tree species was also measured. Mean supercooling point (SCP) of hemlock looper eggs was lower than -30 °C from October through the following spring with values being as low as -47 °C in February. Trehalose was the most abundant sugar found in hemlock looper eggs with a peak concentration of 0.3 µg mg⁻¹ DW⁻¹. Glycerol, a polyol, was more often absent in eggs of the different populations and tree species tested in the study. When exposed to different temperature regimes for various periods of time, significant mortality of hemlock looper eggs occurred at higher temperatures than the mean SCP. Thus, hemlock looper could be considered as a chill tolerant species. No clear pattern of population and host plant effects on SCP and cryoprotectants was detected in this study. However, when exposed to different winter temperatures and exposure duration, hemlock looper from higher latitudes survived better (survival rates ranging between 0 and 89% at -20 °C) than those from lower latitudes (survival rates ranging between 0 and 56% at -20 °C). Our results may contribute to a better understanding of hemlock looper winter biology and thus facilitate predictions of outbreaks and range expansion.

Supercritical CO2 fractionation of bio-oil produced from wheat-hemlock biomass.

The biomass i.e. wheat-hemlock used in this study was first characterized for its composition. The physical and chemical characterization of biomass was estimated using proximate analysis, calorific value, crystallinity, devolatilization behaviour, ultimate analysis, ICP-MS of ash, FT-IR, XRD, CHNS, and HPLC analysis. For commercial purpose the same biomass was used for conversion to bio-oil by fast pyrolysis process. Therefore, in order to investigate its composition, the bio-oil was also characterized using proximate analysis, calorific value, whereas the chemical composition of the bio-oil was estimated using CHNS, (1)H NMR, GC-FID and GC/MS. The bio-oil obtained from wheat-hemlock biomass was supplied by Advanced Biorefinery Co. and after the analysis, its composition has been determined. It contains a mixture of hydrocarbons, pyranoids, furanoids, benzenoids and fatty acids/alcohols with 45% of water, which forms azeotrope with organic polar compounds. The supercritical CO(2) (SC-CO(2)) is an advanced method for selective extraction of valuable chemicals from bio-oil without solvent residue. The organic fraction of the bio-oil was isolated by SC-CO(2). It was observed that SC-CO(2) fractions collected at 10 and 25 MPa pressure were enriched with furanoids, pyranoids and bezenoids. Similarly the bio-oil was also fractionated by conventional column chromatographic method and the yields and chemical compositions were compared with fractionated bio-oil obtained using SC-CO(2).

Highly enantioselective synthesis of chiral cyclic amino alcohols and conhydrine by ruthenium-catalyzed asymmetric hydrogenation.

A highly efficient enantio- and diastereoselective synthesis of chiral cis-beta-N-alkyl/arylamino cyclic alcohols has been realized by asymmetric hydrogenation of racemic alpha-amino cyclic ketones via DKR catalyzed by [RuCl(2)((S)-Xyl-SDP)((R,R)-DPEN)]. The enantioselectivities of the reaction were up to 99.9% ee with 99:1 cis-selectivities. A practical catalytic asymmetric synthesis of all four isomers of conhydrine was also developed.

Differential effects of sugar maple, red oak, and hemlock tannins on carbon and nitrogen cycling in temperate forest soils.

Tannins are abundant secondary chemicals in leaf litter that are hypothesized to slow the rate of soil-N cycling by binding protein into recalcitrant polyphenol-protein complexes (PPCs). We studied the effects of tannins purified from sugar maple, red oak, and eastern hemlock leaf litter on microbial activity and N cycling in soils from northern hardwood-conifer forests of the northeastern US. To create ecologically relevant conditions, we applied tannins to soil at a concentration (up to 2 mg g(-1) soil) typical of mineral soil horizons. Sugar maple tannins increased microbial respiration significantly more than red oak or hemlock tannins. The addition of sugar maple tannins also decreased gross N mineralization by 130% and, depending upon the rate of application, decreased net rates of N mineralization by 50-290%. At low concentrations, the decrease in mineralization appeared to be driven by greater microbial-N immobilization, while at higher concentrations the decrease in mineralization was consistent with the formation of recalcitrant PPCs. Low concentrations of red oak and hemlock tannins stimulated microbial respiration only slightly, and did not significantly affect fluxes of inorganic N in the soil. When applied to soils containing elevated levels of protein, red oak and hemlock tannins decreased N mineralization without affecting rates of microbial respiration, suggesting that PPC formation decreased substrate availability for microbial immobilization. Our results indicate that tannins from all three species form recalcitrant PPCs, but that the degree of PPC formation and its attendant effect on soil-N cycling depends on tannin concentration and the pool size of available protein in the soil.