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Over 1,000 pyrrolizidine alkaloids (PAs) and their N-oxides (PA-N-oxides) occur in 3% of all flowering plants. PA-N-oxides are toxic when reduced to their parent PAs, which are bioactivated into pyrrole intermediates that generate protein- and DNA-adducts resulting in liver toxicity and carcinogenicity. Literature data for senecionine N-oxide in rats indicate that the relative potency (REP) value of this PA-N-oxide compared to its parent PA senecionine varies with the endpoint used. The first endpoint was the ratio between the area under the concentration-time curve (AUC) for senecionine upon dosing senecionine N-oxide or an equimolar dose of senecionine, while the second endpoint was the ratio between the amount for pyrrole-protein adducts formed under these conditions. This study aimed to investigate the mode of action underlying this endpoint dependent REP value for senecionine N-oxide with physiologically based kinetic (PBK) modeling. Results obtained reveal that limitation of 7-GS-DHP adduct formation due to GSH depletion, resulting in increased pyrrole-protein adduct formation, occurs more likely upon high dose oral PA administration than upon an equimolar dose of PAN- oxide. At high dose levels, this results in a lower REP value when based on pyrrole-protein adduct levels than when based on PA concentrations. At low dose levels, the difference no longer exists. Altogether, the results of the study show how the REP value for senecionine N-oxide depends on dose and endpoint used, and that PBK modeling provides a way to characterize REP values for PA-N-oxides at realistic low dietary exposure levels, thus reducing the need for animal experiments. [ABSTRACT FROM AUTHOR]

Background and purpose: This research aimed at evaluating the effects of senecionine and Senecio vulgaris extract on apoptosis and cell cycle inhibition. Also, their cytotoxicity on prostate cancer cell lines (PC3, DU145 and HGFs), antioxidant effects, and influences on AR and CLU gene expression were investigated. Materials and methods: In this experimental study, senecionine and S. vulgaris extract were prepared and the induced morphological changes on cell lines were evaluated in DMEM F12 cell culture after 24, 48, 72, and 96 hours by an invert microscope. Then, cell viability and apoptosis were determined by MTT and fluorescence analysis, respectively. Finally, AR and CLU gene expression and methylation of AR gene were evaluated using Real-time PCR and Bi-Sulfide test, respectively. Results: Cytotoxicity data showed morphological changes 24 hours after treatment with senecionine in all cell lines and all concentrations which also increased in longer time. Based on MTT results, senecionine showed cytotoxicity in 50 and 500 μg/ml on PC3 and du145 prostate cancer cell lines. Conclusion: According to current findings, Ragwort aerial part extract are suggested for in vivo studies with minimum destruction on normal cells and inducing effects on destruction and apoptosis of prostate carcinogenic cell lines.

1,2-unsaturated Pyrrolizidine Alkaloids (PAs) are secondary plant metabolites that occur as food contaminants. Upon consumption, they can cause severe liver damage. PAs have been shown to induce apoptosis, to have cytotoxic and genotoxic effects, and to impair bile acid homeostasis in the human hepatoma cell line HepaRG. The major mode of action of PAs is DNA- and protein-adduct formation. Beyond that, nuclear receptor activation has only been observed for one receptor and two PAs, yielding the possibility that other cellular mediators are involved in PA-mediated toxicity. Here, the mode of action of Senecionine (Sc), a prominent and ubiquitous representative of hepatotoxic PAs, was investigated by analyzing 7 hepatic microRNAs (miRNAs) in HepaRG cells. Ultimately, 11 target genes that were predicted with Ingenuity Pathway Analysis software (IPA) were found to be significantly downregulated, while their assigned miRNAs showed significant upregulation of gene expression. According to IPA, these targets are positively correlated with apoptosis and cellular death and are involved in diseases such as hepatocellular carcinoma. Subsequent antagomiR-inhibition analysis revealed a significant correlation between PA-induced miRNA-4434 induction and P21-Activated Kinase-1 (PAK1) downregulation. PAK1 downregulation is usually associated with cell cycle arrest, suggesting a new function of Sc-mediated toxicity in human liver cells.

Pyrrolizidine alkaloids (PAs) are produced by plants as secondary compounds that are the most widely distributed natural toxins. There have been many cases of human toxicity caused by consumption of toxic plant species, as herbal teas and grain or grain products contaminated with PA-containing seeds have been reported. Companies that produce dried spices and tea leaves should examine the PA level in their products. For the first time in the literature, a simple and inexpensive electrochemical assay based on a single-use sensor was introduced for quantitative determination of senecionine (SEN) in the most frequently contaminated food sources. SEN was immobilized on a pencil graphite electrode surface by the passive adsorption technique. Differential pulse voltammetry (DPV) was used to evaluate the oxidation signal of SEN, which was observed to be around +0.95 V. The oxidation signal was specific to the SEN in the sample, and the current value was proportional to its concentration. The selectivity of our assay was also tested in the presence of other similar PAs such as intermedine, lycopsamine, and heliotrine. The detection limit is calculated by developed assay and found to be 5.45 µg/mL, which is an acceptable concentration value of SEN occurring at toxic levels for consumers. As an application of the developed sensor in food products, the electrochemical detection of SEN was successfully performed in flour and herbal tea products.

Pyrrolizidine alkaloids (PAs) are a large group of botanical toxins of concern, as they are considered genotoxic carcinogens, with long-term dietary exposure presenting an elevated risk of liver cancer. PAs can contaminate honey through honeybees visiting the flowers of PA-containing plant species. A program of monitoring New Zealand honey has been undertaken over several years to build a comprehensive dataset on the concentration, regional and seasonal distribution, and botanical origin of 18 PAs and PA N-oxides. A bespoke probabilistic exposure model has then been used to assess the averaged lifetime dietary risk to honey consumers, with exposures at each percentile of the model characterized for risk using a margin of exposure from the Joint World Health Organization and United Nations Food and Agriculture Organization Expert Committee on Food Additives (JECFA) Benchmark Dose. Survey findings identify the typical PA types for New Zealand honey as lycopsamine, echimidine, retrorsine and senecionine. Regional and seasonal variation is evident in the types and levels of total PAs, linked to the ranges and flowering times of certain plants. Over a lifetime basis, the average exposure an individual will receive through honey consumption is considered within tolerable levels, although there are uncertainties over high and brand-loyal consumers, and other dietary contributors. An average lifetime risk to the general population from PAs in honey is not expected. However, given the uncertainties in the assessment, risk management approaches to limit or reduce exposures through honey are still of value.

Jacobaea aquatica (=Senecio aquaticus, Asteraceae), marsh ragwort, grows regionally in low management intensity wet grasslands in various European countries. The plant can be regarded as a noxious weed as it contains pyrrolizidine alkaloids (PAs), which cause health problems in livestock. In the Waldviertel region of Northern Austria, marsh ragwort has established dense populations as the fertilizing and production intensity of the meadows changed during the last decades. Ragwort biomass production and alkaloid concentration in J. aquatica were recorded during three consecutive years at three sites to study the effects of two treatments: cutting twice a year either early in June plus September, or in July plus August, respectively. Six PAs (Z‐erucifoline > senecionine > integerrimine > seneciphylline > acetyl‐erucifoline and acetyl‐seneciphylline) were evaluated. The alkaloid contents were highest in July and August when 600–1,400 mg/kg dry matter (DM) total PAs occurred in most ragwort samples. Consequently, the forage can easily be contaminated with 40–80 mg/kg DM of PAs and may present a health risk for domestic animals and also affect human health as these alkaloids are transferred into milk. [ABSTRACT FROM AUTHOR]

Pyrrolizidine alkaloids (PAs) are heterocyclic secondary metabolites with a typical pyrrolizidine motif predominantly produced by plants as defense chemicals against herbivores. They display a wide structural diversity and occur in a vast number of species with novel structures and occurrences continuously being discovered. These alkaloids exhibit strong hepatotoxic, genotoxic, cytotoxic, tumorigenic, and neurotoxic activities, and thereby pose a serious threat to the health of humans since they are known contaminants of foods including grain, milk, honey, and eggs, as well as plant derived pharmaceuticals and food supplements. Livestock and fodder can be affected due to PA-containing plants on pastures and fields. Despite their importance as toxic contaminants of agricultural products, there is limited knowledge about their biosynthesis. While the intermediates were well defined by feeding experiments, only one enzyme involved in PA biosynthesis has been characterized so far, the homospermidine synthase catalyzing the first committed step in PA biosynthesis. This review gives an overview about structural diversity of PAs, biosynthetic pathways of necine base, and necic acid formation and how PA accumulation is regulated. Furthermore, we discuss their role in plant ecology and their modes of toxicity towards humans and animals. Finally, several examples of PA-producing crop plants are discussed.

Senecio inaequidensis a neophyte originating from South Africa that has managed to spread to Europe and colonize large areas. This plant has toxic pyrrolizidine alkaloids (PAs) that represent a health risk to humans and animals. The aim of this work was to measure the content of PAs of these plants in continuous intervals over the entire growing season. The plants were separated into their organs, such as sprouts, stems, leaves and inflorescences. PAs were extracted from the dried plants using an acidic methanol solution, purified and measured by GC/MS. The average PA content of plants over the growing season was 0.33% of dry weight. The highest PA levels were about 1% in the dry weight and were found in the young sprouts and flower heads. There were nine PAs present, of which six could be identified. The main alkaloid was retrorsine followed by senecivernine, senecionine, integerrimine, usaramine and seneciphylline. Due to the high PA content of the inflorescences, the long flowering period and the rapid invasion dynamics, this species presents a high health risk for humans and animals. [ABSTRACT FROM PUBLISHER]

Coltsfoot (Tussilago farfara L.) is a common species, widely used in European and Chinese traditional medicine for the treatment of respiratory diseases. However, raw material from this plant contains hepatotoxic pyrrolizidine alkaloids (PAs). The aim of the study was to determine the variability of the level of PAs (senkirkine and senecionine) in leaves of coltsfoot originated from natural populations in Poland. In the phytochemical analysis, 20 samples of T. farfara were used. This plant material was obtained from the Garden of Medicinal Plants in Plewiska near Poznań and originated from different regions of Poland. Coltsfoot leaves were harvested in the middle of July of 2010 and then dried at room temperature. The alkaloid content was detected using the HPLC-DAD method. The amount of PAs in leaves of T. farfara changed in a wide range from 0.06 to 1.04 μg g−1 of dry matter (DM). The content of senkirkine and senecionine was positively correlated (r = 0.68, P = 0.001). There was no statistically significant correlation between the amount of PAs as well as leaf weight and water content in leaves of T. farfara. Our results showed that a medium-sized leaf of coltsfoot (0.33 g DM) may contain from 0.02 to 0.34 μg of PAs (on average 0.14 μg). The level of PAs was not associated with the region of Poland, but phytochemical similarity of samples was usually visible at the local scale. Coltsfoot leaves are characterized by a high variability of the content of toxic PAs, much higher than in the case of the main active compounds, especially flavonoids and mucilage. This phytochemical variability is mainly genetically determined (samples came from a garden collection), and it can be increased by environmental factors. Our investigations indicate that Polish natural populations of T. farfara may provide raw material with a low level of toxic PAs.

Coltsfoot (Tussilago farfara L.) is a common species, widely used in European and Chinese traditional medicine for the treatment of respiratory diseases. However, raw material from this plant contains hepatotoxic pyrrolizidine alkaloids (PAs). The aim of the study was to determine the variability of the level of PAs (senkirkine and senecionine) in leaves of coltsfoot originated from natural populations in Poland. In the phytochemical analysis, 20 samples of T. farfara were used. This plant material was obtained from the Garden of Medicinal Plants in Plewiska near Poznań and originated from different regions of Poland. Coltsfoot leaves were harvested in the middle of July of 2010 and then dried at room temperature. The alkaloid content was detected using the HPLC-DAD method. The amount of PAs in leaves of T. farfara changed in a wide range from 0.06 to 1.04 µg g-1 of dry matter (DM). The content of senkirkine and senecionine was positively correlated (r = 0.68, P = 0.001). There was no statistically significant correlation between the amount of PAs as well as leaf weight and water content in leaves of T. farfara. Our results showed that a medium-sized leaf of coltsfoot (0.33 g DM) may contain from 0.02 to 0.34 µg of PAs (on average 0.14 µg). The level of PAs was not associated with the region of Poland, but phytochemical similarity of samples was usually visible at the local scale. Coltsfoot leaves are characterized by a high variability of the content of toxic PAs, much higher than in the case of the main active compounds, especially flavonoids and mucilage.This phytochemical variability is mainly genetically determined (samples came from a garden collection), and it can be increased by environmental factors. Our investigations indicate that Polish natural populations of T. farfara may provide raw material with a low level of toxic PAs. [ABSTRACT FROM AUTHOR]

1,2-unsaturated Pyrrolizidine Alkaloids (PAs) are secondary plant metabolites that occur as food contaminants. Upon consumption, they can cause severe liver damage. PAs have been shown to induce apoptosis, to have cytotoxic and genotoxic effects, and to impair bile acid homeostasis in the human hepatoma cell line HepaRG. The major mode of action of PAs is DNA- and protein-adduct formation. Beyond that, nuclear receptor activation has only been observed for one receptor and two PAs, yielding the possibility that other cellular mediators are involved in PA-mediated toxicity. Here, the mode of action of Senecionine (Sc), a prominent and ubiquitous representative of hepatotoxic PAs, was investigated by analyzing 7 hepatic microRNAs (miRNAs) in HepaRG cells. Ultimately, 11 target genes that were predicted with Ingenuity Pathway Analysis software (IPA) were found to be significantly downregulated, while their assigned miRNAs showed significant upregulation of gene expression. According to IPA, these targets are positively correlated with apoptosis and cellular death and are involved in diseases such as hepatocellular carcinoma. Subsequent antagomiR-inhibition analysis revealed a significant correlation between PA-induced miRNA-4434 induction and P21-Activated Kinase-1 (PAK1) downregulation. PAK1 downregulation is usually associated with cell cycle arrest, suggesting a new function of Sc-mediated toxicity in human liver cells. [ABSTRACT FROM AUTHOR]

Pyrrolizidine alkaloids (PAs) are considered to be one of the most hepatotoxic groups of compounds of plant origin and are present in about 3% of the world's flowering plants. Most PAs represent a considerable health hazard to both livestock and humans through the consumption of plants and PA-contaminated products such as milk, honey, herbal teas, and medicines. This study determined the differences in the in vivo pharmacokinetic behavior of senecionine (SEN), adonifoline (ADO), and their main metabolites in rats after intravenous administration and oral administration by ultraperformance liquid chromatography/electrospray ionization mass spectrometry. Upon intravenous administration and oral administration of SEN and ADO, significant differences in pharmacokinetics were observed, with the SEN and ADO being absorbed fast with lower bioavailability and being quickly metabolized to PA N-oxides and hydroxylation products of PAs or their N-oxides. It could be seen that the plasma concentration ratio of senecionine N-oxide (SEN-NO) to SEN ( C/ C) was significantly larger than that for adonifoline N-oxide (ADO-NO) and ADO ( C/ C) ( P < 0.001) for both dosing routes in rats. The high N-oxygenation activity and extensive toxicity of SEN, compared with ADO, in rats raised the question of whether or not the higher metabolic rate of SEN in rats in vivo was related to its potent toxicity. The toxicity of SEN-NO and ADO-NO needs to be evaluated further and compared in vitro/in vivo. This study was most helpful for interpreting the metabolism of metabolic bioactivation and detoxication, and toxicity differences among SEN, ADO and other PAs. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Abstract: A rapid, selective and sensitive ultra-performance liquid chromatography–electrospray ionization mass spectrometry (UPLC–ESIMS) method was firstly developed and validated for the simultaneous determination of two hepatotoxic pyrrolizidine alkaloids (HPAs), senecionine (SEN), adonifoline (ADO), and their N-oxides (SENNOX and ADONOX), the main metabolites in rat serum. The whole analysis was achieved within 4.5min by gradient elution on an ACQUITY UPLC BEH C18 column (50mm×2.1mm, i.d. 1.7μm) following a solid phase extraction for serum samples. Good linearity was achieved using weighted (1/x 2) least squares linear regression over a 1600-fold dynamic range for SEN and ADO (LLOQ was about 0.006μg/ml) and 800-fold dynamic range for SENNOX and ADONOX (LLOQ was about 0.012μg/ml). The R.S.D. of intra- and inter-day precision was below 4.91% and 11.15% respectively, while the R.E. of accuracy was within 4.52%, 6.81%, 2.69%, and 7.12% for SEN, SENNOX, ADO, and ADONOX, respectively. The developed method was successfully applied to the in vivo pharmacokinetic study in rats after intravenous administration of SEN and ADO. [Copyright &y& Elsevier]

Abstract: Pyrrolizidine alkaloids (PAs) are well-known natural hepatotoxins distributed widely in thousands of plants in the world. Adonifoline (Adon), senecionine (Sene) and monocrotaline (Mono) are retronecine-type PAs, and the present study is designed to observe the effects of intracellular glutathione on toxicity of these three PAs in human normal liver L-02 cells. The ratio of cellular reduced glutathione (GSH) and oxidized glutathione (GSSG) was assayed after L-02 cells were incubated with these three PAs for various times. Results showed that Adon, Sene and Mono all significantly decreased the ratio of GSH/GSSG in L-02 cells in the time- and concentration-dependent manner. The results of 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) and trypan blue staining assay showed that these three PAs all significantly decreased cell viability in L-02 cells when pretreated with 10μM BSO (L-Buthionine-S-R-Sulfoximine) for 24h to deplete intracellular GSH. Further results showed that anti-oxidant compounds such as NAC (N-Acetyl-Cysteine) and GSH could rescue the cytotoxicity caused by these three PAs with BSO pretreatment. Taken together, those results suggest that intracellular GSH plays important roles in regulating the cytotoxicity induced by PAs. [Copyright &y& Elsevier]

Abstract: Gas chromatography–mass spectrometry analysis of seeds from 28 species of Crotalaria from Brazil (sections Calycinae, Crotalaria, Chrysocalycinae and Hedriocarpae) showed that pyrrolizidine alkaloids (PAs) are important as chemotaxonomic markers at the infrageneric level. The sections Calycinae and Crotalaria were characterized by 11-membered macrocyclic monocrotaline-type PAs. In the section Chrysocalycinae, a single species in the subsection Incanae, C. incana, showed integerrimine, a 12-membered macrocyclic senecionine-type, as main PA. The group close to the subsection Stipulosae (C. micans and C. maypurensis) showed distinctive PA patterns: C. micans presented mainly the 12-membered macrocyclic integerrimine, and C. maypurensis the unusual 7-hydroxy-1-methylene-8-pyrrolizidine. In the group close to the subsection Glaucae, the PAs with otonecine as the necine base were the main alkaloids, except in C. rufipila which showed an assamicadine-like PA (monocrotaline-type). The section Hedriocarpae showed main 12-membered macrocyclic senecionine-type PAs. [Copyright &y& Elsevier]

A rapid reversed-phase high-performance liquid chromatographic (RP-HPLC) method was established for simultaneous determination of senecionine, senlciphylline, and senecionine N-oxide in a famous traditional Chinese medicine, Gynura segetum, which has been commonly used for hemostasis. The HPLC assay was performed on a Kromasil KR100-5 C18 column (25 cm × 4.6 mm, 5 µm) with mobile phase composed of acetonitrile and 0.2% phosphoric acid-triethylamine within 40 min. The detection wavelength was 220 nm. All the compounds showed good linearity (r2 > 0.9997). The method was reproducible with intra- and interday variation less than 2.82%. The recovery of the assay was in the range of 96.55-103.88%. The method was successfully applied to the quantification of three constituents in 15 Gynura segetum samples collected from different metropolis. The results indicated that the developed assay could be considered as a suitable quality-control method for Gynura segetum. [ABSTRACT FROM AUTHOR]

Abstract: Tussilago farfara (Kuan Donghua) is an important Chinese herbal medicine which has been shown to contain many bioactive compounds and widely used to relieve cough and resolve phlegm. However, besides therapeutic bioactive compounds, this herb has been found to contain toxic pyrrolizidine alkaloids (PAs), mainly senkirkine and traces of senecionine. In this report, conditions for microwave-assisted extraction (MAE) and pressurized hot water extraction (PHWE) were optimized for the extraction of the PAs. The results were compared against heating under reflux. It was found that the binary mixture of MeOH:H2O (1:1) acidified using HCl to pH 2–3 was the optimal solvent for the extraction of the PAs in the plant materials. Liquid chromatography (LC) with ultra-violet (UV) detection and electrospray ionization mass spectrometry (ESI-MS) in the positive mode was used for the determination and quantitation of senkirkine and senecionine in the botanical extract. The proposed extraction methods with LC/MS allow for the rapid detection of the major and the minor alkaloids in T. farfara in the presence of co-eluting peaks. With LC/MS, the quantitative analysis of PAs in the extract was done using internal standard calibration and the precision was found to vary from 0.6% to 5.4% on different days. The limits of detection (LODs) and limits of quantitation (LOQs) for MAE and PHWE were found to vary from 0.26μg/g to 1.04μg/g and 1.32μg/g to 5.29μg/g, respectively. The method precision of MAE and PHWE were found to vary from 3.7% to 10.4% on different days. The results showed that major and minor alkaloids extracted using MAE and PHWE were comparable to that by heating under reflux. Our data also showed that significant ion suppression was not observed in the analysis of senkirkine and senecionine in the botanical extracts with co-eluting peaks. [Copyright &y& Elsevier]

Pyrrolizidine alkaloids (PAs) are a large group of botanical toxins of concern, as they are considered genotoxic carcinogens, with long-term dietary exposure presenting an elevated risk of liver cancer. PAs can contaminate honey through honeybees visiting the flowers of PA-containing plant species. A program of monitoring New Zealand honey has been undertaken over several years to build a comprehensive dataset on the concentration, regional and seasonal distribution, and botanical origin of 18 PAs and PA N-oxides. A bespoke probabilistic exposure model has then been used to assess the averaged lifetime dietary risk to honey consumers, with exposures at each percentile of the model characterized for risk using a margin of exposure from the Joint World Health Organization and United Nations Food and Agriculture Organization Expert Committee on Food Additives (JECFA) Benchmark Dose. Survey findings identify the typical PA types for New Zealand honey as lycopsamine, echimidine, retrorsine and senecionine. Regional and seasonal variation is evident in the types and levels of total PAs, linked to the ranges and flowering times of certain plants. Over a lifetime basis, the average exposure an individual will receive through honey consumption is considered within tolerable levels, although there are uncertainties over high and brand-loyal consumers, and other dietary contributors. An average lifetime risk to the general population from PAs in honey is not expected. However, given the uncertainties in the assessment, risk management approaches to limit or reduce exposures through honey are still of value. [ABSTRACT FROM AUTHOR]

Pyrrolizidine alkaloids (PAs) are produced by plants as secondary compounds that are the most widely distributed natural toxins. There have been many cases of human toxicity caused by consumption of toxic plant species, as herbal teas and grain or grain products contaminated with PA-containing seeds have been reported. Companies that produce dried spices and tea leaves should examine the PA level in their products. For the first time in the literature, a simple and inexpensive electrochemical assay based on a single-use sensor was introduced for quantitative determination of senecionine (SEN) in the most frequently contaminated food sources. SEN was immobilized on a pencil graphite electrode surface by the passive adsorption technique. Differential pulse voltammetry (DPV) was used to evaluate the oxidation signal of SEN, which was observed to be around +0.95 V. The oxidation signal was specific to the SEN in the sample, and the current value was proportional to its concentration. The selectivity of our assay was also tested in the presence of other similar PAs such as intermedine, lycopsamine, and heliotrine. The detection limit is calculated by developed assay and found to be 5.45 µg/mL, which is an acceptable concentration value of SEN occurring at toxic levels for consumers. As an application of the developed sensor in food products, the electrochemical detection of SEN was successfully performed in flour and herbal tea products. [ABSTRACT FROM AUTHOR]

Abstract: The contents and the compositions of the pyrrolizidine alkaloid (PA) complex of ragwort (Senecio jacobaea L.) were examined as potential drivers of fungal community structure in the rhizosphere. S. jacobaea plants within the coastal sand dune reserve of Meijendel (the Netherlands) were assayed for concentration and composition of PAs in roots. Rhizosphere soil was collected from pre-flowering plants, which differed up to 8-fold in PA production, and represented both jacobine and senecionine/seneciphylline chemotypes. Bulk soil samples from the same site were also collected for comparative examination. A culture-independent approach, involving direct DNA isolation, PCR of fungal 18S rRNA genes, and denaturing gradient gel electrophoresis (DGGE), was applied to compare the fungal communities of plants with different PA contents, as well as differences between bulk and rhizosphere samples. Cluster analysis of PCR-DGGE profiles revealed no clear evidence for PA-induced selection of specific fungal communities. However, canonical variance analysis showed that fungal communities associated with high-PA jacobine chemotypes could be discriminated from low PA samples and from the senecionine/seneciphylline chemotypes. The diversity of DGGE banding patterns, both in terms of band number and evenness, showed a trend toward lower diversity in the rhizosphere of high-PA plants as compared to low-PA plants and bulk soil. These results indicate that PA chemotypes of S. jacobaea differ in their influence on soil-borne fungal communities, with jacobine-containing plants exerting a greater selection in the rhizosphere than plants containing senecionine/seneciphylline. [Copyright &y& Elsevier]

Pyrrolizidine alkaloids (PAs) are known to protect Arctiidae moths and Danainae and Ithomiinae butterflies against the orb-weaving spider Nephila clavipes (Araneae, Araneidae, Tetragnathinae), which liberates adults of these insects unharmed from its web. We tested against this spider the role of eight PAs and one derived structure [an 89:11 mixture of the 12-membered macrocyclic diester free base integerrimine and senecionine and the respective N-oxide; two hydrolysis products from this mixture (the necine base retronecine, its respective N-oxide, and a mixture of integerriminic and senecionic necic acids); the 12-membered macrocyclic senkirkine; the 9- O-monoester free base senecioylretronecine and its respective N-oxide; and the 9- O-monoester free base callimorphine (a PA biosynthesized only by insects from retronecine)]. The mixture integerrimine–senecionine N-oxide seems to be more active than the respective free base [LibD50 (liberation dose 50) = 0.042 and 0.153 μg/dry weight of prey, respectively], but the difference in activity between the N-oxide and free base of the 9- O-monoester senecioylretronecine was slight (LibD50 = 0.167 and 0.104, respectively). Senkirkine, an otonecine base PA that does not form N-oxide and is not found in insects, was the less active, showing the highest LibD50 (0.354). The difference in antipredator activity between N-oxides and free bases from macrocyclic diesters and monoesters may be correlated with physicochemical properties of these molecules in interaction with the Nephila receptors. For the active structures, there was a significant correlation between dosage and antipredator activity. Both forms of retronecine and a necic acid mixture were inactive, supporting the hypothesis that PAs biosynthesized by insects from retronecine were originally produced and stored in order to optimize chemical defense. Comparison of dose/activity data with reported amounts of PAs in butterflies suggested that, in general, PA-specialist insects are protected against predation by Nephila. [ABSTRACT FROM AUTHOR]

Hepatic sinusoidal obstruction syndrome ( HSOS), characterized by hepatomegaly, ascites and hyperbilirubinemia, is caused by toxic injury to hepatic sinusoidal endothelial cells. One major etiology of HSOS in China is the intake of products containing pyrrolizidine alkaloids ( PA) such as Tusanqi. The manifestations of patients with HSOS are usually non-specific, presenting with abnormal liver function and portal hypertension. Diagnosis of the disease depends mostly on liver histopathology when clinical and imaging data are not sufficient. A history of Tusanqi intake is mostly important for the diagnosis. Due to a lack of effective, evidence-based treatments for HSOS, avoiding the mistaken use of PA-containing products including Tusanqi is important for the prevention of HSOS. [ABSTRACT FROM AUTHOR]

Pyrrolizidine alkaloids (PAs) are heterocyclic secondary metabolites with a typical pyrrolizidine motif predominantly produced by plants as defense chemicals against herbivores. They display a wide structural diversity and occur in a vast number of species with novel structures and occurrences continuously being discovered. These alkaloids exhibit strong hepatotoxic, genotoxic, cytotoxic, tumorigenic, and neurotoxic activities, and thereby pose a serious threat to the health of humans since they are known contaminants of foods including grain, milk, honey, and eggs, as well as plant derived pharmaceuticals and food supplements. Livestock and fodder can be affected due to PA-containing plants on pastures and fields. Despite their importance as toxic contaminants of agricultural products, there is limited knowledge about their biosynthesis. While the intermediates were well defined by feeding experiments, only one enzyme involved in PA biosynthesis has been characterized so far, the homospermidine synthase catalyzing the first committed step in PA biosynthesis. This review gives an overview about structural diversity of PAs, biosynthetic pathways of necine base, and necic acid formation and how PA accumulation is regulated. Furthermore, we discuss their role in plant ecology and their modes of toxicity towards humans and animals. Finally, several examples of PA-producing crop plants are discussed. [ABSTRACT FROM AUTHOR]