Poisoning in ruminants caused by species of the genus Cestrum L. (Solanaceae) in Brazil: A review of toxicological and phytochemical evidence.

Some plant species of the genus Cestrum L. (Solanaceae family) are known to cause poisoning in farming animals in Brazil, negatively affecting the livestock sector. In this context, this study aimed to carry out a systematic review of the Cestrum species that cause poisoning in ruminants in Brazil and to list the main phytochemicals involved in these toxic activities that have already been identified. Scientific documents were retrieved in Google Scholar, PubMed®, ScienceDirect®, and SciELO databases. After applying the inclusion criteria, a total of 38 articles published between 1920 and 2023 were included in the present study. Cestrum axillare Vell. [Syn. Cestrum laevigatum Schltdl.], Cestrum corymbosum Schltdl., Cestrum intermedium Sendtn., and Cestrum parqui L'Hér. were found to have reported cases of poisoning in the Northeast, Southeast, and South of Brazil. Natural poisonings in ruminants caused by these species have been recorded in ten Brazilian states, mostly in Rio de Janeiro, Santa Catarina, Rio Grande do Sul, and Pernambuco. In general, Cestrum species cause liver damage and a clinical-pathological state characterized by acute liver failure of the poisoned animals. Cattle are more susceptible to poisoning by these plants, but there are reports of poisoning by C. axillare in goats and buffaloes as well. Several chemical constituents were identified in C. axillare and C. parqui, including some saponins and terpenoids that may be associated with the cases of poisoning. However, only one chemical compound has been identified in C. intermedium, and no phytochemical investigation has been carried out regarding toxic compounds in C. corymbosum. It is expected that future studies fill the gap in determining the toxic principles present in these species.

Essential Oil of Cestrum diurnum L.: GC/MS Analysis, in Vitro and in Silico Anti-HCoV-229E Effects and Inhibitory Activity against LPS-Induced Inflammation.

Cestrum diurnum L. (Solanaceae) is a fragrant ornamental tree cultivated in different parts around the world. In this study, the essential oil (EO) of the aerial parts was extracted by hydrodistillation (HD), steam distillation (SD) and microwave-assisted hydro-distillation (MAHD). GC/MS analysis of the three EOs revealed that phytol represents the major component in SD-EO and MAHD-EO (40.84 and 40.04 %, respectively); while in HD-EO it only represented 15.36 %. The SD-EO showed a strong antiviral activity against HCoV-229E with IC50 of 10.93 µg/mL, whereas, MAHD-EO and HD-EO showed a moderate activity with IC50 values of 119.9 and 148.2 µg/mL, respectively. The molecular docking of EO major components: phytol, octadecyl acetate and tricosane showed a strong binding to coronavirus 3-CL (pro). Moreover, the three EOs (50 µg/mL) decreased the levels of NO, IL-6 and TNF-α and suppressed IL-6 and TNF-α gene expression in LPS-induced inflammation model in RAW264.7 macrophage cell lines.

Production and bioprocessing of camptothecin from Aspergillus terreus, an endophyte of Cestrum parqui, restoring their biosynthetic potency by Citrus limonum peel extracts.

The metabolic potency of fungi as camptothecin producer elevates their prospective use as an industrial platform for commercial production, however, the loss of camptothecin productivity by fungi with the storage and subculturing are the major obstacle. Thus, screening for endophytic fungal isolates inhabiting ethnopharmacological plants with an obvious metabolic stability and sustainability for camptothecin biosynthesis could be one of the most feasible paradigms. Aspergillus terreus ON908494.1, an endophyte of Cestrum parqui was morphologically and molecularly verified, displaying the most potent camptothecin biosynthetic potency. The chemical identity of A. terreus camptothecin was confirmed from the HPLC, FTIR and LC-MS/MS analyses, gave the same molecular structure and mass fragmentation patterns of authentic one. The purified putative camptothecin displayed a strong anticancer activity towards HepG-2 and MCF-7 with IC50 values 0.96 and 1.4 µM, respectively, with no toxicity to OEC normal cells. As well as, the purified camptothecin displayed a significant antifungal activity towards fungal human pathogen Candida albicans, Aspergillus flavus, and A. parasiticus, ensuring the unique structural activity relationships of A. terreus camptothecin, as a powerful dually active anticancer and antimicrobial agent. The camptothecin productivity of A. terreus was maximized by bioprocessing with Plackett-Burman design, with an overall 1.5 folds increment (170.5 µg/L), comparing to control culture. So, the optimal medium components for maximum yield of camptothecin by A. terreus was acid why (2.0 mL/L), Diaion HP20 (2.0 g/L), Amberlite XAD (2.0 g/L), dextrin (5.0 g/L), glucose (10.0 g/L), salicylic acid (2.0 g/L), serine (4.0 g/L), cysteine (4.0 g/L) and glutamate (10.0 g/L), at pH 6 for 15 days incubation. By the 5th generation of A. terreus, the camptothecin yield was reduced by 60%, comparing to zero culture. Interestingly, the productivity of camptothecin by A. terreus has been completely restored and over increased (210 µg/L), comparing to the 3rd generation A. terreus (90 µg/L) upon addition of methanolic extracts of Citrus limonum peels, revealing the presence of some chemical signals that triggers the camptothecin biosynthetic machinery. The feasibility of complete restoring of camptothecin biosynthetic-machinery of A. terreus for stable and sustainable production of camptothecin, pave the way for using this fungal isolate as new platform for scaling-up the camptothecin production.

Cestrum axillare (Solanaceae) poisoning in ruminants.

Cestrum axillare poisoning causes significant economic losses in farms of ruminant production due to a fatal acute hepatic disease. The consumption of C. axillare occurs on farms or pastures with a scarcity of feed or with dry forage. Epidemiological, clinical, and pathological data of poisoning outbreaks by C. axillare from 1953 to 2021 in grazing ruminants in southeastern Brazil are reported. A total of 68 bovines, two buffaloes, and two goats exhibited clinical signs and resulted in death due to C. axillare consumption, with 79% of the cases occurring during the dry period. Clinical signs were apathy, anorexia, ruminal arrest, arched back, and constipation with hard stools, sometimes with blood or mucus. Cases with neurological signs due to hepatic encephalopathy showed excitement, aggressiveness, drooling, staggering, and muscle tremors. The pathological findings included hepatocellular necrosis in the liver and microcavitations in the brain's white matter (status spongiosus). The hepatotoxins, carboxyparquin and parquin, were detected in C. axillare leaf samples collected from paddocks grazed by cattle in three southeastern Brazilian municipalities where outbreaks of C. axillare poisoning occurred. This is the first report of parquin and carboxyparquin in C. axillare.

Network pharmacology-based study on apigenin present in the methanolic fraction of leaves extract of Cestrum nocturnum L. to uncover mechanism of action on hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the sixth most common type of cancer in the world. It is the third leading cause of cancer-related mortality. In more than 80% of people liver cancer-related death is due to its poor prognosis. The flavonoids obtained from natural sources have potent therapeutic effects on HCC. The flavonoid rich methanolic fraction obtained from ethyl acetate extract of leaf of Cestrum nocturnum (MFLCN) was analyzed by UPLC-QTOFMS/MS for the presence of different flavonoids. The physiochemical and pharmacokinetics properties of the identified flavonoids were performed by absorption, distribution, metabolism, excretion, and toxicity (ADMET). It was selected on the basis of Lipinski rule and hepatotoxicity evaluations. The potential gene dataset of HCC were taken from gene card database and targets compounds were selected from target net prediction. Gene ontology and pathway enrichment analysis of HCC was performed via enricher and David web tools. Cytoscape was used to visualize targets and network pathways. MFLCN contains 33 flavonoids. Among these flavonoids, apigenin was selected as principal active compound on the basis of their pharmacokinetic and ADMET properties. Apigenin has 92 targets out of 627 total targets related to HCC, while there was13 pathways in the target-pathway network. Results revealed that apigenin regulates cell proliferation and survival, primarily through different signaling pathways like estrogen, VEGF, PI3K/AKT1, TNF, FoXO, and Ras signaling pathways. Thus, integrating network pharmacology prediction with m-RNA and human protein atlas validation could be an effective method for understanding the molecular mechanism of apigenin on HCC.

Diversity of the repetitive DNA fraction in Cestrum, the genus with the largest genomes within Solanaceae.

BACKGROUND: Cestrum species present large genomes (2 C = ~ 24 pg), a high occurrence of B chromosomes and great diversity in heterochromatin bands. Despite this diversity, karyotypes maintain the chromosome number 2n = 16 (except when they present B chromosomes), and a relative similarity in chromosome morphology and symmetry. To deepen our knowledge of the Cestrum genome composition, low-coverage sequencing data of C. strigilatum and C. elegans were compared, including cytogenomic analyses of seven species. METHODS AND RESULTS: Bioinformatics analyses showed retrotransposons comprising more than 70% of the repetitive fraction, followed by DNA transposons (~ 17%), but FISH assays using retrotransposon probes revealed inconspicuous and scattered signals. The four satellite DNA families here analyzed represented approximately 2.48% of the C. strigilatum dataset, and these sequences were used as probes in FISH assays. Hybridization signals were colocalized with all AT- and GC-rich sequences associated with heterochromatin, including AT-rich Cold-Sensitive Regions (CSRs). Although satellite probes hybridized in almost all tested species, a satDNA family named CsSat49 was highlighted because it predominates in centromeric regions. CONCLUSIONS: Data suggest that the satDNA fraction is conserved in the genus, although there is variation in the number of FISH signals between karyotypes. Except to the absence of FISH signals with probes CsSat1 and CsSat72 in two species, the other satellites occurred in species of different phylogenetic clades. Some satDNA sequences have been detected in the B chromosomes, indicating that they are rich in preexisting sequences in the chromosomes of the A complement. This comparative study provides an important advance in the knowledge on genome organization and heterochromatin composition in Cestrum, especially on the distribution of satellite fractions between species and their importance for the B chromosome composition.

Isolation and Identification of Pennogenin Tetraglycoside from Cestrum nocturnum (Solanaceae) and Its Antifungal Activity against Fusarium kuroshium, Causal Agent of Fusarium Dieback.

Antifungal assay-guided fractionation of the methanolic crude extract of Cestrum nocturnum (Solanaceae), popular known as 'lady of the night', led the isolation and identification of the steroidal saponin named pennogenin tetraglycoside, which was identified for the first time in this plant species by spectroscopic means. The crude extract, fractions and pennogenin tetraglycoside exhibited mycelial growth inhibition of Fusarium solani and F. kuroshium. F. solani is a cosmopolitan fungal phytopathogen that affects several economically important crops. However, we highlight the antifungal activity displayed by pennogenin tetraglycoside against F. kuroshium, since it is the first plant natural product identified as active for this phytopathogen. This fungus along with its insect symbiont known as Kuroshio shot hole borer (Euwallacea kuroshio) are the causal agents of the plant disease Fusarium dieback that affects more than 300 plant species including avocado (Persea americana) among others of ecological relevance. Scanning electron microscopy showed morphological alterations of the fungal hyphae after exposure with the active fractions and 12 phenolic compounds were also identified by mass spectrometry dereplication as part of potential active molecules present in C. nocturnum leaves.

Sensing of mercury ion using light induced aqueous leaf extract mediated green synthesized silver nanoparticles of Cestrum nocturnum L.

In this study, a simple, one-pot, and eco-friendly biosynthesis of silver nanoparticles (AgNPs) was accomplished with the use of aqueous leaves extract of Cestrum nocturnum L.(AECN). Different techniques like ultraviolet-visible (UV-Vis) spectrophotometry, Fourier transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning area electron diffraction were used to investigate the optical, operational, and physical properties of the green synthesized AECN-AgNPs.The AECN-AgNPs were further used for the detection of Hg2+ by UV-Vis and electrochemical methods. The disintegration of the AECN-AgNPs solution caused the formation of an Ag-Hg amalgam, which caused discoloration of the solution. Sensing performance for a variety of metals such as Na+, K+, Mg2+, Ca2+, Ni2+, Cu 2+, Fe3+, Zn2+, Co2+, Cd2+, Pb2+, As3+, and Mn2+ at 10-mM concentrations was measured in order to determine the selectivity of the sensor towards the Hg2+. For the electrochemical determination of 2 + Hg2+ , AECN-AgNPs were immobilized on a glassy carbon (GC) electrode, and the resulting modified electrode (GC/AECN-AgNPs) was characterized by cyclic voltammetry. This phenomenon is advantageously used for the sensitive determination of trace level Hg2+. GC/AECN-AgNPs demonstrated a linear calibration range of 100 nM to 10 µM and a limit of detection of 21 nM for Hg2+ determination.

Green Synthesized Zinc Oxide Nanoparticles Based on Cestrum diurnum L. of Potential Antiviral Activity against Human Corona 229-E Virus.

SARS-CoV-2 has caused more than 596 million infections and 6 million fatalities globally. Looking for urgent medication for prevention, treatment, and rehabilitation is obligatory. Plant extracts and green synthesized nanoparticles have numerous biological activities, including antiviral activity. HPLC analysis of C. dirnum L. leaf extract showed that catechin, ferulic acid, chlorogenic acid, and syringic acid were the most major compounds, with concentrations of 1425.16, 1004.68, 207.46, and 158.95 µg/g, respectively. Zinc nanoparticles were biosynthesized using zinc acetate and C. dirnum extract. TEM analysis revealed that the particle size of ZnO-NPs varied between 3.406 and 4.857 nm. An XRD study showed the existence of hexagonal crystals of ZnO-NPs with an average size of 12.11 nm. Both ZnO-NPs (IC50 = 7.01 and CC50 = 145.77) and C. dirnum L. extract (IC50 = 61.15 and CC50 = 145.87 µg/mL) showed antiviral activity against HCOV-229E, but their combination (IC50 = 2.41 and CC50 = 179.23) showed higher activity than both. Molecular docking was used to investigate the affinity of some metabolites against the HCOV-229E main protease. Chlorogenic acid, solanidine, and catchin showed high affinity (-7.13, -6.95, and -6.52), compared to the ligand MDP (-5.66 Kcal/mol). Cestrum dinurum extract and ZnO-NPs combination should be subjected to further studies to be used as an antiviral drug.

Biosystematic studies of some Egyptian species of Cestrum (Solanaceae).

Cestrum is the second largest genus of family Solanaceae, after Solanum, distributed in warm to subtropical regions. Species of genus Cestrum are one of the most ethnopharmacological relevant plants, for their broad biological and pharmacological properties. There is a scarcity to taxonomical studies and identification of these plants in Egypt, thus, the objective of this study was to implement various morphological features, chemical markers and molecular tools to emphasize the taxonomical features of the different Cestrum species. Morphologically, the epidermal cells of C. diurnum, C. elegans and C. parqui were irregular with sinuate anticlinal wall patterns for both surfaces, while, C. nocturnum has anticlinal walls, sinuolate with polygonal to irregular epidermal cells on the abaxial surface. The species of Cestrum have hypostomatic leaves, except C. parqui that has amphistomatic leaves. The experimented species of Cestrum have Anomocytic and anisocytic stomata, while, C. elegans has a diacytic stomata. The morphologically identified Cestrum spp were molecular confirmed based on their ITS sequences, the sequences of C. diurnum, C. nocturnum, C. elegans and C. parqui were deposited on genbank with accession # MT742788.1, MT749390.1, MW091481.1 and MW023744.1, respectively. From the SCOT analyses, the four species of Cestrum were grouped into 2 clusters (I, II), cluster I contains C. elegans, C. nocturnum and C. parqui, while cluster II contains only C. diurnum with 100% polymorphism for all primers. From the GC-MS profile, the C. diurnum exhibited a diverse metabolic paradigm, ensuring their richness with different metabolites comparing to other experimented Cestrum species. Among the total resolved metabolites, 15-methyltricyclo 6.5.2-pentadeca-1,3,5,7,9, 11,13-heptene was the highly incident compound in C. elegans (35.89%) followed by C. parqui (21.81%) and C. diurnum (11.28%), while it absent on C. nocturnum. The compound, 2,2',6,6'-tetra-tert-butyl-4,4'-methylenediphenol was highly detected in C. elegans and C. dirunum with minor amounts in the other Cestrum species. Cypermethrin and 3-butynyl-2,2,5-trimethyl-1,3-dioxane-5-methanol were pivotally reported in C. nocturnum. Taken together, from molecular and metabolic markers, C. diurnum, C. parqui and C. elegans have higher proximity unlike to C. nocturnum.

Anatomical and genetic bases underlying the convergent evolution of fleshy and dry dehiscent fruits in Cestrum and Brugmansia (Solanaceae).

The mechanisms controlling evolutionary shifts between dry and fleshy fruits in angiosperms are poorly understood. In Solanaceae, Cestrum and Brugmansia represent cases of convergent evolution of fleshy and dry fruits, respectively. Here we study the anatomical and genetic bases of the independent origin of fleshy fruits in Cestrum and the reversion to dry dehiscent fruits in Brugmansia. We also characterize the expression of candidate fruit development genes, including ALCATRAZ/SPATULA, FRUITFULL, HECATE1/2/3, REPLUMLESS and SHATTERPROOF. We identify anatomical changes to establish developmental stages in the ovary-to-fruit transition in Cestrum nocturnum and Brugmansia suaveolens. We generate reference transcriptomes for both species, isolate homologs for all genes in the fruit genetic regulatory network (GRN) and perform gene expression analyses for ALC/SPT, FUL, HEC1/2/3, RPL and SHP throughout fruit development. Finally, we compare our results to expression patterns found in typical capsules of Nicotiana tabacum and berries of Solanum lycopersicum available in public repositories. We have identified homologous, homoplasious and unique anatomical features in C. nocturnum and B. suaveolens fruits, resulting in their final appearance. Expression patterns suggest that FUL, SHP and SPT might control homologous characteristics, while ALC and RPL likely contribute to homoplasious anatomical features. The fruit GRN changes considerably in these genera when compared to typical capsules and berries of Solanaceae, particularly in B. suaveolens, where expression of FUL2 and RPL1 is lacking.

Spontaneous poisoning by Cestrum intermedium in dairy cattle / Intoxicação espontânea por Cestrum intermedium em bovinos de leite

Acute hepatotoxicity caused by plants poisoning is responsible for economic losses in farm animals in Brazil. Reports of Cestrum intermedium natural poisoning in cattle are not commonly described in Rio Grande do Sul (RS). This study aimed to document an outbreak of spontaneous C. intermedium poisoning in dairy cattle in the Central-Eastern Mesoregion of RS. Three nine-month-old Holstein and Jersey heifers were affected after they were placed in a small paddock with shortage forage. In this area, specimens of C. intermedium Sendtn with signs of consumption were observed. Morbidity and lethality rates were 100% and clinical courses ranged from 9 to 12 hours. At post mortem examination of the three heifers, there was predominance of acute liver lesions. The liver was moderately enlarged and on the cut surface there was a marked accentuation of the lobular pattern and hemorrhage. Inside the rumen, partially digested C. intermedium Sendtn leaves were observed. The histological aspects of the liver were mostly centrilobular coagulative necrosis and hemorrhage, frequently extended to the midzonal region. The immunohistochemistry technique was performed, in which the polyclonal antibody caspase 3 was used in liver fragments. Moderate to marked immunolabeling was observed in the cytoplasm and nucleus of hepatocytes, predominantly on the periphery of areas of hepatic necrosis indicating cell apoptosis. The diagnosis of C. intermedium Sendtn poisoning in dairy cattle in this study was based on epidemiological, clinical and anatomopathological findings. Since the C. intermedium poisoning is uncommon in dairy cattle, we are describing it for the first time in the Central-Eastern Mesoregion of RS, and represents a differential diagnosis of other acute toxic liver diseases in cattle.(AU)

Intoxicações por plantas que causam hepatotoxicidade aguda são responsáveis por prejuízos econômicos em animais de produção no Brasil. Relatos de intoxicações naturais por Cestrum intermedium em bovinos não são comumente descritos no Rio Grande do Sul (RS). Este trabalho teve como objetivo documentar um surto de intoxicação espontânea por C. intermedium em bovinos leiteiros ocorrido na região Centro-Oriental do RS. Três bovinos com nove meses de idade, das raças Holandês e Jersey, foram acometidos depois de serem introduzidos em um pequeno piquete com escassez de forragem. Nessa área foram observados exemplares de Cestrum intermedium Sendtn com sinais de consumo. As taxas de morbidade e letalidade foram de 100% e os cursos clínicos variaram de 9 a 12 horas. Na necropsia dos três bovinos, observou-se predomínio de lesões hepáticas agudas. O fígado dos animais acometidos estava moderadamente aumentado de tamanho e na superfície de corte havia marcada acentuação do padrão lobular e hemorragia. Na abertura do rúmen, foram observadas folhas de Cestrum intermedium Sendtn parcialmente digeridas. Histologicamente, o fígado apresentava necrose coagulativa centrolobular e hemorragia. Frequentemente, essas alterações se estendiam para a região mediozonal. Foi realizada a técnica de imuno-histoquímica, na qual utilizou-se o anticorpo policlonal caspase 3 em fragmentos de fígado. Notou-se marcação moderada a acentuada em citoplasma e núcleo de hepatócitos, predominantemente na periferia das áreas de necrose hepática indicando apoptose celular. O diagnóstico de intoxicação por Cestrum intermedium Sendtn nos bovinos leiteiros desse estudo, foi realizado com base nos achados epidemiológicos, clínicos e anatomopatológicos. A intoxicação por C. intermedium é incomum em bovinos leiteiros, sendo descrita pela primeira vez na região centro oriental do RS e representa um diagnóstico diferencial de outras hepatopatias tóxicas agudas.(AU)

A Total of Eight Novel Steroidal Glycosides Based on Spirostan, Furostan, Pseudofurostan, and Cholestane from the Leaves of Cestrum newellii.

Previously, various steroidal glycosides were reported from plants of Cestrum species. However, phytochemical investigation has not been conducted on Cestrum newellii. A systematic phytochemical investigation of the leaves of C. newellii resulted in the isolation of eight novel steroidal glycosides (1-8), which were classified into three spirostanol glycosides (1-3), two furostanol glycosides (4 and 5), two pseudofurostanol glycosides (6 and 7), and one cholestane glycoside (8). In addition, three known cholestane glycosides (9-11) were isolated and identified. The structures of the new compounds were determined based on spectroscopic data and chemical transformations. Compounds 1 and 2 are spirostanol glycosides having hydroxy groups at C-2, C-3, C-12, and C-24 of the aglycone moiety. Although C. newellii is known to be a poisonous plant, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay exhibited that none of the isolated compounds were cytotoxic to HL-60 human promyelocytic leukemia cells.

IGS sequences in Cestrum present AT- and GC-rich conserved domains, with strong regulatory potential for 5S rDNA.

The 35S and 5S ribosomal DNA (rDNA) organized in thousands of copies in genomes, have been widely used in numerous comparative cytogenetic studies. Nevertheless, several questions related to the diversity and organization of regulatory motifs in 5S rDNA remain to be addressed. The 5S rDNA unit is composed of a conserved 120 bp length coding region and an intergenic spacer (IGS) containing potential regulatory motifs (Poly-T, AT-rich and GC-rich) differing in number, redundancy and position along the IGS. The Cestrum species (Solanaceae) have large genomes (about 10 pg/1C) and conserved 2n = 16 karyotypes. Strikingly, these genomes show high diversity of heterochromatin distribution, variability in 35S rDNA loci and the occurrence of B chromosomes. However, the 5S rDNA loci are highly conserved in the proximal region of chromosome 8. Comparison of seventy-one IGS sequences in plants revealed several conserved motifs with potential regulatory function. The AT- and GC-rich domains appeared highly conserved in Cestrum chromosomes. The 5S genic and the GC-rich IGS probe produced FISH signals in both A (pair 8) and B chromosomes. The GC-rich domain presented a strong potential for regulation because it may be associated with CpG islands organization, as well as to hairpin and loop organization. Another interesting aspect was the ability of AT- and GC-rich motifs to produce non-heterochromatic CMA/DAPI signals. While the length of the 5S rDNA IGS region varied in size between the Cestrum species, the individual sequence motifs seem to be conserved suggesting their regulatory function. The most striking feature was the conserved GC-rich domain in Cestrum, which is recognized as a signature trait of the proximal region of chromosome pair 8.

In vitro anticancer and antioxidant potential of Cestrum species.

The methanolic extract of leaves and stem of Cestrum nocturnum and Cestrum diurnum were investigated for their antioxidant and anticancer attribute through standard methods. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was carried out to estimate the antioxidant activity of the extracts. Whereas, anticancer potential of extracts were tested against colon cancer cell line, HCT 116 and acute myeloid leukemia (AML) cell lines, THP-1. Results showed that extracts of both plants exhibited a very strong antioxidant activity in a dose dependent manner. In addition, both extracts efficiently increased the cell death in two different cancer cell lines. Moreover, DNA fragmentation analysis further strengthens the anticancer potential of extracts of both types of plants. Current study, therefore, provide a preliminary data highlighting the antioxidant and anticancer activities of methanolic extract of leaves and stem of Cestrum nocturnum and Cestrum diurnum.

Hypolipidemic effect of steroid compound from Cestrum diurnum (Solanaceae: Solanales) in normocholesterolemic albino rat.

The present study was carried out to establish the hypolipidemic effect of a phyto-steroid compound isolated from the chloroform: methanol extract of fresh mature leaves of the plant Cestrum diurnum (Solanaceae: Solanales). Change in the haematological parameters was studied in normocholesterolemic albino rats after oral administration of single dose of isolated phytosteroid (2 mg/ day) up to 15 days and compared with control rats. Application of phytosteroid fraction resulted in a significant reduction in total plasma cholesterol and free cholesterol levels. The plasma triglyceride levels also decreased significantly. A sharp increase in the HDL cholesterol level and a significant decrease in the LDL and VLDL amount were also documented. Free fatty acid level was significantly low in treated rats.

Comparação dos efeitos das folhas de Cestrum axillare Vell. com as saponinas isoladas em caprinos / Comparison of the effects of Cestrum axillare leaves with isolated saponins in goats

Cestrum axillare Vell. (formerly Cestrum laevigatum Schltd.), family Solanaceae, is the most important hepatotoxic plant in Brazil that causes acute poisoning. It occurs in the Southeast and Center-West regions and in coastal areas of the Northeast Brazil. Spontaneous poisoning was described in cattle, goats and sheep, with clinical signs evidenced within 24 hours after ingestion of the leaves and death within 48 hours after signs onset. The clinical signs observed in acute poisoning are apathy, anorexia, ruminal arrest, arched back, constipation with feces in small spheres, sometimes covered with mucus and blood streaks, muscle tremors, staggering gait and sometimes sialorrhoea. Neurological signs may be observed, due to interference in the urea cycle due to hepatic insufficiency resulting in hyperammonemia (hepatic encephalopathy). The main pathological finding is centrilobular hepatic necrosis. The toxic principle present in C. axillare was not yet definitively proven, but some authors attribute the toxicity of the plant to the presence of saponins gitogenin and digitogenin. However, it has not been determined whether the saponins present in C. axillare are responsible for the hepatotoxic effect of the plant. Thus, the objective of this work is to determine if the saponins are the compounds responsible for the hepatotoxic effects produced by the ingestion of the leaves of C axillare, using goats as experimental model. For this, the effects of the administration of the leaves were compared with those produced by the saponins isolated from the leaves in goats. Six goats were randomly assigned to three experimental groups that received [1] dry leaves of C. axillare (animals A1 and A2), [2] saponins extract from leaves (animals S1 and S2) or [3] control group (animals C1 and C2). For goats receiving the dry leaves the administered dose of plant was 10g/kg for one animal (A1) and 5g/kg for the other one (A2). For animals receiving the saponins extract, administration was done at a dose equivalent to 20g/kg repeated after 24 hours. The dry leaves administered at a dose of 10g/kg to a goat produced toxic effects, with alterations in biochemistry (indicating hepatic lesion) and histopathology showing centrilobular hepatic necrosis. At the dose of 5 g/kg of dry leaves, clinical signs of poisoning were not observed, but hepatic necrosis was found; after 15 days after the last administration, the hepatic parenchyma of this animal was already normal, with only hemorrhagic areas, demonstrating full regeneration. The administration of extracts of saponins containing gitogenin and digitogenin to goats did not produce significant toxic effects, proving that these compounds are not responsible for intoxication. In addition, goats are a good experimental model for studies of this intoxication.(AU)

Cestrum axillare Vell. (anteriormente C. laevigatum Schltd.), família Solanaceae, é a mais importante planta hepatotóxica do Brasil que causa intoxicação aguda. Tem ocorrência nas regiões Sudeste e Centro-Oeste e em áreas litorâneas do Nordeste. A intoxicação natural foi descrita em bovinos, caprinos e ovinos, com sinais clínicos evidenciados em até 24 horas após a ingestão das folhas e morte em até 48 horas após o início da sintomatologia. Os sinais clínicos observados na intoxicação aguda são apatia, anorexia, parada ruminal, dorso arqueado, constipação com fezes em formas de pequenas esferas, por vezes recobertas com muco e com estrias de sangue, tremores musculares, andar cambaleante e, às vezes, sialorreia. Podem ser observados sinais neurológicos, devido à interferência no ciclo da ureia pela insuficiência hepática resultando em hiperamonemia (encefalopatia hepática). O principal achado patológico é a necrose hepática centrolobular. O princípio tóxico presente no C. axillare ainda não está definitivamente comprovado, mas alguns autores atribuem a toxicidade da planta à presença das saponinas gitogenina e digitogenina. No entanto, ainda não foi determinado se as saponinas presentes em C. axillare são as responsáveis pelo efeito hepatotóxico da planta. Assim, o objetivo deste trabalho é determinar se as saponinas são os compostos responsáveis pelos efeitos hepatotóxicos produzidos pela ingestão das folhas de C. axillare, usando caprinos como modelo experimental. Para isto, foram comparados os efeitos da administração das folhas com os produzidos pelas saponinas isoladas destas folhas em caprinos. Foram utilizados seis caprinos, distribuídos aleatoriamente em três grupos experimentais que receberam [1] folhas secas de C. axillare (Caprinos A1 e A2), [2] extrato de saponinas das folhas (Caprinos S1 e S2), e [3] grupo controle (Caprinos C1 e C2). Para os caprinos que receberam as folhas secas a dose administrada de planta foi de 10g/kg para um animal (A1) e de 5g/kg para outro (A2). Para os animais que receberam o extrato de saponinas, a administração foi feita na dose equivalente a 20g/kg, repetida após 24 horas. Foi verificado que as folhas secas, quando administradas na dose de 10g/kg a um caprino, produziram efeitos tóxicos, com alterações na bioquímica (indicando lesão hepática) e histopatológica apresentando necrose hepática centrolobular. Na dose de 5g/kg de folhas secas, não foi observado sintomatologia clínica da intoxicação, mas houve necrose hepática; 15 dias após a última administração, o parênquima hepático deste animal já se encontrava normal, apenas com áreas hemorrágicas, demonstrando plena regeneração. A administração do extrato de saponinas contendo gitogenina e digitogenina a caprinos não produziu efeitos tóxicos significantes, comprovando não serem estes compostos os responsáveis pela intoxicação. Além disto, a espécie caprina é um bom modelo experimental para estudos desta intoxicação.(AU)

BAL31-NGS approach for identification of telomeres de novo in large genomes.

This article describes a novel method to identify as yet undiscovered telomere sequences, which combines next generation sequencing (NGS) with BAL31 digestion of high molecular weight DNA. The method was applied to two groups of plants: i) dicots, genus Cestrum, and ii) monocots, Allium species (e.g. A. ursinum and A. cepa). Both groups consist of species with large genomes (tens of Gb) and a low number of chromosomes (2n=14-16), full of repeat elements. Both genera lack typical telomeric repeats and multiple studies have attempted to characterize alternative telomeric sequences. However, despite interesting hypotheses and suggestions of alternative candidate telomeres (retrotransposons, rDNA, satellite repeats) these studies have not resolved the question. In a novel approach based on the two most general features of eukaryotic telomeres, their repetitive character and sensitivity to BAL31 nuclease digestion, we have taken advantage of the capacity and current affordability of NGS in combination with the robustness of classical BAL31 nuclease digestion of chromosomal termini. While representative samples of most repeat elements were ensured by low-coverage (less than 5%) genomic shot-gun NGS, candidate telomeres were identified as under-represented sequences in BAL31-treated samples.

Usos y costumbres sobre hierbas medicinales en barrios periurbanos del sudeste de la ciudad de Salta, Argentina / Uses and customs on medicinal herbs in periurban neighborhoods ofsoutheastern Salta City, Argentina

Desde la medicina oficial se desconoce y subestima el uso de otras formas de atención, por lo que este trabajo tiene como objetivos caracterizar el conocimiento sobre uso de hierbas medicinales en la zona sudeste de la ciudad de Salta; identificar cuáles se utilizan para tratar problemas de salud; y valorar su frecuencia de uso. Se realizó una investigación cuali-cuantitativa, entrevistando a informantes claves y grupos focales. Se confeccionaron listas de hierbas medicinales, y sus usos se categorizaron por problemas de salud. Se realizó una encuesta sobre frecuencia de usos medicinales. Se mencionaron 59 hierbas medicinales con una media de 2,3 usos por planta. Con un "Nivel de importancia Significativo" se encuentran quimpe, tusca, paico, coca, molle, ruda y hediondilla. El "coqueo" se encontró en 54% de las familias. El 52% utiliza alguna hierba para afecciones gastrointestinales, el 45% para problemas de la piel, el 40 % para afecciones respiratorias, el 33% para "empacho" y 17% para problemas renales, sin diferencias significativas en los usos por grupo etáreo. La población tiene amplio conocimiento sobre hierbas medicinales. La medicina formal debe legitimar estos conocimientos e incluirlos en su práctica.

Traditional health professionals underestimate and fail to recognize the use of other forms of health care, therefore this research has the objective of characterizing knowledge about the use of medicinal herbs in the southeastern area of Salta city; to identify which herbs are used to treat health problems; and to assess its frequency of use. Qualitative and quantitative research was conducted by interviewing key informants and focus groups. Lists of medicinal herbs and uses were created and categorized by health problems. Finally, a closed survey on frequency of use was carried out. Fifty-nine herbs were mentioned with an average of 2.3 uses per plant. quimpe, tusca , paico, coca, molle, ruda and hediondillawere herbs marked as “with a Significant Level of Impotanse”. The use of coca was found in 54% of the families interviewed. Fifty-two percent used some herb for gastrointestinal disorders, 45% for skin problems, 40% for respiratory conditions, 33% for indigestion and 17% for kidney problems, with no significant differences in uses according toage groups. Population has a good knowledge of medicinal herbs. Formal medicine should legitimize this knowledge and include it in its practice.

Spirostanol glucosides from the leaves of Cestrum laevigatum L.

Two new steroidal saponins, (25R)-spirost-5-ene-3ß,26ß-diol 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-[(1 → 2)-α-L-rhamnopyranosyl]-ß-D-glucopyranoside (1) and (25R)-spirost-6-ene-3ß,5ß-diol 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-[(1 → 2)-α-L-rhamnopyranosyl]-ß-D-glucopyranoside (2), along with the known diosgenin 3-O-α-L-rhamnopyranosyl-(1 → 4)-α-L-rhamnopyranosyl-(1 → 4)-ß-D-glucopyranoside (3), chonglouoside SL-5 (4) and Paris saponin Pb (5) were isolated from the leaves of Cestrum laevigatum. The structures of the compounds were determined using spectroscopic analyses including HRESI-MS, 1D and 2D NMR data, followed by comparison with data from the literature. Among them, two are particularly unique, compound 1 is the first (6)Δ-spirostanol saponin and compound 2 has an unusual C-26 hydroxyl in the (5)Δ-spirostanol skeleton. Antifungal testing showed a potent activity to formosanin C against Candida albicans and Candida parapsilosis. Evaluation of the cytotoxic activity indicated that compound 1 has a moderate activity against HL-60 and SF-295 cell lines, while compound 2 were active only against HL-60.