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1.
Am J Emerg Med ; 54: 329.e1-329.e3, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34776281

RESUMO

Acute angle closure glaucoma is a sight-threatening condition that may lead to blindness. This is a case report of a woman who presented to the emergency department (ED) with acute angle closure glaucoma following use of an over-the-counter (OTC) homeopathic eye drop containing atropa belladonna (deadly nightshade). A 55-year-old woman presented to the ED with a 5-day history of left eye redness, swelling, tearing, and foreign-body sensation that had acutely worsened in the last two days. Her exam revealed mild left conjunctival injection with watery tearing and a hazy appearance of her left cornea. Fluorescein staining was negative, while tonometry revealed elevated intraocular pressure on the left, suggestive of acute angle closure glaucoma. She was urgently referred to ophthalmology. The etiology of the acute angle closure glaucoma was initially unclear however, with additional prompting, she revealed that two days prior she had started using homeopathic OTC eye drops. Inspection of the eyedrop's ingredients revealed that atropa belladonna was the primary ingredient and likely precipitated her isolated episode of acute angle closure glaucoma. A high level of clinical suspicion and focused ophthalmic exam including tonometry is essential to identify acute angle closure glaucoma in the ED. We present a case report of acute angle closure glaucoma associated with the use of homeopathic belladonna-containing eyedrops. Our report reinforces the necessity to perform thorough medication and supplement history given the prevalence of physiologically active substances available in OTC medications.


Assuntos
Atropa belladonna , Glaucoma de Ângulo Fechado , Glaucoma , Feminino , Glaucoma/induzido quimicamente , Glaucoma/tratamento farmacológico , Glaucoma de Ângulo Fechado/induzido quimicamente , Glaucoma de Ângulo Fechado/tratamento farmacológico , Humanos , Pressão Intraocular , Pessoa de Meia-Idade , Soluções Oftálmicas/efeitos adversos
2.
Clin Toxicol (Phila) ; 60(2): 252-254, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33988064

RESUMO

BACKGROUND: A 53-year-old male with no pre-existing conditions and no permanent medication presented to our emergency department with an anticholinergic syndrome including confusion, anxiety, ataxia and dysarthria after ingestion of a homeopathic solution containing Atropa belladonna extract supposedly in a D4 dilution. METHODS: Atropine sulphate was quantitatively analysed in serum and the homeopathic preparation via liquid chromatography/mass spectrometry. RESULTS: Analysis revealed concentrations of approximately 3 mg/mL atropine sulphate in the homeopathic solution and a serum level of 5.7 ng/mL (±1.4) in the patient's blood proving a 600-fold overdose of atropine due to a production error of the homeopathic dilution. The patient was observed and recovered without further intervention. CONCLUSION: Rare but possibly dangerous manufacturing errors should be considered when faced with symptoms occurring after ingestion of homeopathic or holistic remedies.


Assuntos
Síndrome Anticolinérgica , Atropa belladonna , Síndrome Anticolinérgica/etiologia , Síndrome Anticolinérgica/terapia , Atropa belladonna/química , Atropina/uso terapêutico , Humanos , Masculino , Pessoa de Meia-Idade , Antagonistas Muscarínicos , Extratos Vegetais/química
3.
Nat Prod Res ; 36(5): 1370-1374, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33459053

RESUMO

Alkaloid profiles from Amaryllis belladonna plants collected in Chile were examined by GC-MS to assess their inhibitory activity on acetylcholinesterase (AChE) using in vitro and in silico methodologies. The alkaloid extract was roughly separated by column chromatography on silica gel. AChE inhibitory activities from extracts and purified alkaloids were tested by the Ellman method and a molecular docking study was performed to assess the interaction between AChE and purified alkaloids. Sixteen alkaloids were found from hexane and chloroform extracts, and three were isolated and identified as buphanidrine, acetylcaranine and lycorine. Chloroform extract showed the greatest AChE inhibitory activity with IC50 value 8.89 µg/mL, whereas buphanidrine exhibited the highest inhibitory activity, with IC50 value 17.56 µg/mL. Inhibition kinetics showed that buphanidrine acts as a mixed inhibitor and molecular docking supports this inhibition mechanism. Overall, our study supports the potential use of A. belladonna as an alkaloid source with AChE inhibitory activity.[Formula: see text].


Assuntos
Acetilcolinesterase , Atropa belladonna , Acetilcolinesterase/química , Chile , Inibidores da Colinesterase/química , Simulação de Acoplamento Molecular , Extratos Vegetais/química
4.
Biosci Biotechnol Biochem ; 85(12): 2404-2409, 2021 Nov 24.
Artigo em Inglês | MEDLINE | ID: mdl-34562094

RESUMO

Tropane alkaloids, including clinically important hyoscyamine and scopolamine, are produced in the roots of medicinal plant species, such as Atropa belladonna, from the Solanaceae family. Recent molecular and genomic approaches have advanced our understanding of the metabolic enzymes involved in tropane alkaloid biosynthesis. A noncanonical type III polyketide synthase, pyrrolidine ketide synthase (PYKS) catalyzes a two-step decarboxylative reaction, which involves imine-ketide condensation indispensable to tropane skeleton construction. In this study, we generated pyks mutant A. belladonna hairy roots via CRISPR/Cas9-mediated genome editing and analyzed the metabolic consequences of the loss of PYKS activity on tropane alkaloids, providing insights into a crucial role of the scaffold-forming reaction in the biosynthetic pathway.


Assuntos
Atropa belladonna
5.
Physiol Plant ; 172(4): 2098-2111, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33942310

RESUMO

Hyoscyamine (HYO) and scopolamine (SCO) are tropane alkaloids acting as anticholinergic factors on the parasympathetic nervous system in humans and are produced by Solanaceous plants. Two strains of Agrobacterium rhizogenes, A4 and LBA9402, were used to infect Atropa acuminata Royle ex Miers and Atropa belladonna L. leaf explants. A. acuminata was inoculated either by direct infection or sonicated-assisted A. rhizogenes-mediated transformation (SAAT) was performed. A. belladonna was inoculated with the A4 strain using a direct method. The selected hairy root lines of both species were elicited with 50 mM methyl-ß-cyclodextrin (ß-CD), 0.5 µM coronatine (Cor) or 50 mM ß-CD + 0.5 µM Cor on Day 14 of culture. The elicitor effect on growth and HYO and SCO content was analyzed after one (T1) and two (T2) weeks of treatment. In A. acuminata explants, the highest transformation percentage (T%) was obtained with strain A4 and the SAAT method (T%: 96.43). Cor significantly reduced the growth of A. acuminata hairy roots (fresh weight and dry weight [DW]: 2.52 and 0.3 g, respectively), whereas ß-CD increased their DW (0.4 g). Also, the combined ß-CD + Cor treatment had a positive significant effect on the DW of A. belladonna hairy roots (0.41 g). In A. acuminata hairy roots, the HYO level was lower under Cor treatment than in the control at both sampling times. In contrast, the SCO content was increased 10-fold by Cor elicitation at T1 compared to the control (10.95 mg g-1 DW) and was also positively affected by ß-CD + Cor. In A. belladonna hairy roots, all the elicitors had a negative effect on both HYO and SCO production. This report is the first assessment of the effect of ß-CD and Cor elicitors on tropane alkaloid production.


Assuntos
Atropa belladonna , Atropa , Agrobacterium , Aminoácidos , Indenos , Raízes de Plantas , Tropanos , beta-Ciclodextrinas
6.
Appl Biochem Biotechnol ; 193(6): 1654-1674, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33620666

RESUMO

Suitable recognition of invasive microorganisms is a crucial factor for evoking a strong immune response that can combat the pathogen. Toll-like receptors (TLRs) play a pivotal role in the induction of this innate immune response through stimulation of interferons (IFNs) that control viral replication in the host via distinct signaling pathways. Though the antiviral property of Atropa belladonna has been established, yet the role of one of its active components scopolamine in modulating various factors of the innate immune branch has not yet been investigated until date. Thus, the present study was conducted to assess the antiviral effects of scopolamine and its immunomodulatory role against Japanese encephalitis virus (JEV) infections in embryonated chick. Pre-treatment with scopolamine hydrobromide showed a significant decrease in the viral loads of chorioallantoic membrane (CAM) and brain tissues. Molecular docking analysis revealed that scopolamine hydrobromide binds to the active site of non-structural protein 5 (NS5) that has enzymatic activities required for replication of JEV, making it a highly promising chemical compound against the virus. The binding contributions of different amino acid residues at or near the active site suggest a potential binding of this compound. Pre-treatment with the scopolamine hydrobromide showed significant upregulation of different TLRs like TLR3, TLR7, and TLR8, interleukins like IL-4, and IL-10, as well as IFNs and their regulatory factors. However, virus-infected tissues (direct infection group) exhibited higher TLR4 expression as compared to scopolamine hydrobromide pre-treated, virus-infected tissues (medicine pre-treated group). These results indicate that scopolamine hydrobromide contributes much to launch antiviral effects by remoulding the TLR and IFN signaling pathways that are involved in sensing and initiating the much-needed anti-JEV responses.


Assuntos
Proteínas Aviárias/metabolismo , Vírus da Encefalite Japonesa (Espécie)/metabolismo , Encefalite Japonesa/tratamento farmacológico , Escopolamina/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Atropa belladonna/química , Embrião de Galinha , Encefalite Japonesa/metabolismo , Escopolamina/química
7.
Int J Mol Sci ; 22(4)2021 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-33572199

RESUMO

Atropa belladonna L. is one of the most important herbal plants that produces hyoscyamine or atropine, and it also produces anisodamine and scopolamine. However, the in planta hyoscyamine content is very low, and it is difficult and expensive to independently separate hyoscyamine from the tropane alkaloids in A. belladonna. Therefore, it is vital to develop A. belladonna plants with high yields of hyoscyamine, and without anisodamine and scopolamine. In this study, we generated A. belladonna plants without anisodamine and scopolamine, via the CRISPR/Cas9-based disruption of hyoscyamine 6ß-hydroxylase (AbH6H), for the first time. Hyoscyamine production was significantly elevated, while neither anisodamine nor scopolamine were produced, in the A. belladonna plants with homozygous mutations in AbH6H. In summary, new varieties of A. belladonna with high yields of hyoscyamine and without anisodamine and scopolamine have great potential applicability in producing hyoscyamine at a low cost.


Assuntos
Atropa belladonna/metabolismo , Hiosciamina/biossíntese , Engenharia Metabólica/métodos , Oxigenases de Função Mista/genética , Proteínas de Plantas/metabolismo , Atropa belladonna/genética , Atropina/biossíntese , Sistemas CRISPR-Cas/genética , Edição de Genes/métodos , Regulação da Expressão Gênica de Plantas , Técnicas de Inativação de Genes , Hiosciamina/isolamento & purificação , Oxigenases de Função Mista/metabolismo , Mutagênese , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Escopolamina/metabolismo , Sementes/genética , Alcaloides de Solanáceas/biossíntese
8.
J Biotechnol ; 328: 23-33, 2021 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-33422568

RESUMO

Curcumin has ignited global interest as an elite drugable molecule, owing to its time-honoured pharmacological activities against diverse human ailments. Limited natural accessibility and poor oral bioavailability caused major hurdles in the curcumin-based drug development process. We report the first successful testimony of curcumin and its glucoside synthesis in Atropa belladonna hairy roots (HR) through metabolic engineering. Re-routing the inherent biosynthetic precursors of the phenylpropanoid pathway of A. belladonna by heterologous expression of key curcumin biosynthetic pathway genes (i.e., Diketide-CoA synthase-DCS and Curcumin synthase-CURS3) and glucosyltransferase gene (CaUGT2) resulted in the production of curcumin and its glucoside in HR clones. Under shake-flask cultivation, the PGD2-HR1clone bearing DCS/ CURS3 genes showed the maximum curcumin yield (180.62 ± 4.7 µg/g DW), while the highest content of curcumin monoglucoside (32.63 ± 2.27 µg/g DW) along with curcumin (67.89 ± 2.56 µg/g DW) were noted in the PGD3-HR3 clone co-expressing DCS/CURS3 and CaUGT2 genes. Bioreactor up-scaling showed yield improvements in the PGD2-HR1 (2.3 fold curcumin) and the PGD3-HR3 clone (0.9 and 1.65 folds of curcumin-monoglucoside and curcumin respectively). These findings proved the advantageous use of HR cultures as the production source for curcumin and its glucoside, which remained unexplored so far.


Assuntos
Atropa belladonna , Curcumina , Glucosídeos , Humanos , Raízes de Plantas , Biologia Sintética
9.
Nat Prod Res ; 35(7): 1090-1096, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31303055

RESUMO

The culture broth of endophytic Streptomyces sp. AB100, isolated from the shoots of medicinal plant Atropa belladonna (L.) was investigated for the presence of antibacterial compounds. After initial testing followed by bioactivity-guided fractionation, six new piperazic acid (PA)-containing congeners of two known peptides, JBIR-39 and JBIR-40, were identified by HR-MS/MS and NMR analyses. Only the dehydroxylated hexapeptidic derivatives with unusual incorporation of four PA moieties exhibited weak antibacterial activity against Gram-positive test organism Bacillus subtilis. A 16S rDNA-based phylogenetic tree of known Streptomyces spp. producing PA-containing hexapeptides isolated from different habitats and endophyte Streptomyces AB100 showed considerable diversity, suggesting that these metabolites may play an important environmental role beyond their antibacterial activity.


Assuntos
Atropa belladonna/microbiologia , Endófitos/química , Peptídeos/farmacologia , Plantas Medicinais/química , Piridazinas/farmacologia , Streptomyces/química , Streptomyces/isolamento & purificação , Antibacterianos/farmacologia , Bacillus subtilis/efeitos dos fármacos , DNA Ribossômico/genética , Espectroscopia de Ressonância Magnética , Testes de Sensibilidade Microbiana , Filogenia , Brotos de Planta/microbiologia , Espectrometria de Massas em Tandem
10.
Biomed Chromatogr ; 35(5): e5053, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33314218

RESUMO

Atropine is an antimuscarinic alkaloid identified in Atropa belladonna. In pharmacopeias, percolation is standardized as an extraction method for A. belladonna leaves, along with liquid-liquid extraction as a cleanup procedure and titration as an analytical method for assaying the atropine in the leaves. In this study, a faster, solvent-saving, and more reliable method for quality control of A. belladonna samples was developed. Ultrasound-assisted extraction was proposed and optimized by fractional factorial design followed by Box-Behnken design. For modeling atropine content, the following optimal conditions were established: particle size, 180 µm; percentage methanol in water, 50%; volume of solvent, 15 ml; time of extraction, 60 min; and number of extractions, two. This led to a significant improvement in atropine extraction (P < 0.001). For cleanup, solid-phase extraction was used as an alternative to liquid-liquid extraction, giving similar results, with higher reproducibility. Finally, for the atropine assay, a UPLC method was validated as a substitute for the classic titration method. Taken together, the development of an ultrasound-assisted extraction-solid-phase extraction-UPLC approach allowed the determination of atropine content in A. belladonna leaves in a time- and solvent-saving manner, with high reliability.


Assuntos
Atropa belladonna/química , Atropina/análise , Cromatografia Líquida de Alta Pressão/métodos , Extratos Vegetais/química , Folhas de Planta/química , Extração em Fase Sólida/métodos , Antagonistas Muscarínicos/análise , Solventes/química
11.
ACS Chem Neurosci ; 11(21): 3683-3696, 2020 11 04.
Artigo em Inglês | MEDLINE | ID: mdl-33054164

RESUMO

Japanese encephalitis virus (JEV) is the main cause of viral encephalitis resulting in more than 68 000 clinical cases every year with case fatality rate as high as 30-40% for which no specific treatments are available. We have recently exhibited belladonna may be widely applicable for the treatment of various neurological disorders. Therefore, we developed a hydroalcoholic formulation of belladonna (B200) consisting of atropine and scopolamine and showed its antiviral efficacy against JEV infection. B200 treatment increases neuronal cell survival by reducing JEV induced cytopathic effects which were evident from significant reduction in necrotic cell population by flow-cytometry analysis and caspase 3 and 8 enzymatic activities. B200 treatment was found to reduce the intracellular JEV level observed by significant reduction in JEV-fluorescein isothiocyanate (FITC) expression in both neurons and microglia. Because microglia plays a crucial role in JEV pathogenesis, we further investigated the anti-JEV effects of B200 on human microglia cells and elucidated the mechanism of action by performing whole-transcriptome sequencing. Gene expression analysis revealed that B200 reduces the pro-apoptotic and inflammatory gene expression observed by significant reduction in BAD, BAX, CASP3, CASP8, IL1B, and CXCL10 and increase in IL10 responsive gene expression. Interestingly, our molecular docking analysis revealed that atropine and scopolamine interact with the His288 residue of NS3 protein, a crucial residue for RNA unwinding and ATPase activity that was further confirmed by degradation of NS3 protein. Drug likeness, ADME (absorption, distribution, metabolism, and excretion), and toxicity analysis further suggests that atropine and scopolamine both cross the blood-brain barrier, which is crucial for effective treatment of Japanese encephalitis (JE).


Assuntos
Atropa belladonna , Vírus da Encefalite Japonesa (Espécie) , Encefalite Japonesa , Antivirais/farmacologia , Antivirais/uso terapêutico , Sobrevivência Celular , Encefalite Japonesa/tratamento farmacológico , Humanos , Inflamação/tratamento farmacológico , Microglia , Simulação de Acoplamento Molecular , Neurônios
12.
Nature ; 585(7826): 614-619, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32879484

RESUMO

Tropane alkaloids from nightshade plants are neurotransmitter inhibitors that are used for treating neuromuscular disorders and are classified as essential medicines by the World Health Organization1,2. Challenges in global supplies have resulted in frequent shortages of these drugs3,4. Further vulnerabilities in supply chains have been revealed by events such as the Australian wildfires5 and the COVID-19 pandemic6. Rapidly deployable production strategies that are robust to environmental and socioeconomic upheaval7,8 are needed. Here we engineered baker's yeast to produce the medicinal alkaloids hyoscyamine and scopolamine, starting from simple sugars and amino acids. We combined functional genomics to identify a missing pathway enzyme, protein engineering to enable the functional expression of an acyltransferase via trafficking to the vacuole, heterologous transporters to facilitate intracellular routing, and strain optimization to improve titres. Our integrated system positions more than twenty proteins adapted from yeast, bacteria, plants and animals across six sub-cellular locations to recapitulate the spatial organization of tropane alkaloid biosynthesis in plants. Microbial biosynthesis platforms can facilitate the discovery of tropane alkaloid derivatives as new therapeutic agents for neurological disease and, once scaled, enable robust and agile supply of these essential medicines.


Assuntos
Alcaloides/biossíntese , Alcaloides/provisão & distribuição , Hiosciamina/biossíntese , Saccharomyces cerevisiae/metabolismo , Escopolamina/metabolismo , Aciltransferases/genética , Aciltransferases/metabolismo , Animais , Atropa belladonna/enzimologia , Derivados da Atropina/metabolismo , Transporte Biológico , Datura/enzimologia , Glucosídeos/biossíntese , Glucosídeos/metabolismo , Hiosciamina/provisão & distribuição , Lactatos/metabolismo , Ligases/genética , Ligases/metabolismo , Modelos Moleculares , Doenças do Sistema Nervoso/tratamento farmacológico , Oxirredutases/genética , Oxirredutases/metabolismo , Engenharia de Proteínas , Saccharomyces cerevisiae/genética , Escopolamina/provisão & distribuição , Vacúolos/metabolismo
13.
Anal Bioanal Chem ; 412(27): 7405-7416, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32851457

RESUMO

Alkaloids represent a major group of natural products (NPs), derived from highly diverse organisms. These structurally varied specialized metabolites are widely used for medicinal purposes and also known as toxic contaminants in agriculture and dietary supplements. While the detection of alkaloids is generally facilitated by GC- or LC-MS, these techniques do require considerable efforts in sample preparation and method optimization. Bypassing these limitations and also reducing experimental time, matrix-free laser desorption ionization (LDI) and related methods may provide an interesting alternative. As many alkaloids show close structural similarities to matrices used in matrix-assisted laser desorption ionization (MALDI), they should ionize upon simple laser irradiation without matrix support. With this in mind, the current work presents a systematic evaluation of LDI properties of a wide range of structurally diverse alkaloids. Facilitating a direct comparison between LDI and ESI-MS fragmentation, all tested compounds were further studied by electrospray ionization (ESI). Moreover, crude plant extracts of Atropa belladonna, Cinchona succirubra, and Colchicum autumnale were analyzed by LDI in order to evaluate direct alkaloid detection and dereplication from complex mixtures. Finally, dose-dependent evaluation of MALDI and LDI detection using an extract of Rosmarinus officinalis spiked with atropine, colchicine, or quinine was conducted. Overall, present results suggest that LDI provides a versatile analytical tool for analyzing structurally diverse alkaloids as single compounds and from complex mixtures. It may further serve various potential applications ranging from quality control to the screening for toxic compounds as well as the build up of MS databases. Graphical abstract.


Assuntos
Alcaloides/análise , Atropa belladonna/química , Cinchona/química , Colchicum/química , Extratos Vegetais/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
14.
Zhongguo Zhong Yao Za Zhi ; 45(2): 321-330, 2020 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-32237314

RESUMO

Atropa belladonna seedlings were used as experimental materials and cultivated by soil culture method. Different concentrations(0,0.05,0.1,0.2,0.5 mmol·L~(-1))of NO donor sodium nitroprusside(SNP) were sprayed on the leaves. The effects of different concentrations of SNP and different treatment time(4,8,12,16 d) on nitrogen metabolism, secondary metabolite content, precursor content of tropane alkaloid synthesis pathway and expression of key enzyme genes under 100 mmol·L~(-1) NaCl stress were studied. The results showed that with the prolongation of salt stress, the nitrogen metabolism and the accumulation of secondary metabolites of A. belladonna were inhibited to some extent. After treatment with different concentrations of exogenous SNP, the ammonium nitrogen content decreased dramatically, and the contents of nitrate nitrogen, free amino acid, soluble protein and the activities of key enzymes of nitrogen metabolism(NR, GS, GDH) were all greatly improved; the contents of precursor amino acids(ornithine, arginine) and polyamines(Put, Spd, Spm) in the secondary metabolic pathway have increased to varying degrees. The qRT-PCR analysis showed that exogenous SNP treatment can effectively promote the high expression of key enzyme genes PMT, TRⅠ and H6H in the secondary metabolic pathway of A. belladonna, and the production of hyoscyamine and scopolamine were increased notably. In summary, the application of appropriate concentration of SNP can effectively alleviate the inhibition of salt stress on the nitrogen metabolism and secondary metabolism of Atropa belladonna, and enhance its salt tolerance. Overall, 0.1 mmol·L~(-1) and 0.2 mmol·L~(-1) SNP treatment achieved the most remarkable effect.


Assuntos
Atropa belladonna/metabolismo , Hiosciamina/análise , Nitrogênio/metabolismo , Escopolamina/análise , Nitroprussiato , Metabolismo Secundário , Cloreto de Sódio , Estresse Fisiológico
15.
Plant Cell Rep ; 39(7): 921-939, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32300886

RESUMO

KEY MESSAGE: Atropa acuminata aqueous leaf extract biosynthesized silver nanoparticles showed strong antioxidant, anticancerous (HeLa cells) and anti-inflammatory activities. Besides, this bio syn-AgNP also proved effective against mosquito vectors causing malaria, dengue and filariasis. Present study highlights eco-friendly and sustainable approach for the synthesis of silver nanoparticles (AgNP) using aqueous leaf extract of A. acuminata, a critically endangered medicinal herb. The addition of 1 mM silver nitrate to aqueous leaf extract resulted in the synthesis of AgNP when solution was heated at 60 °C for 30 min at pH 7. Absorption band at 428 nm, as shown by UV-Vis spectroscopy confirmed the synthesis of AgNP. XRD patterns revealed the crystalline nature of AgNP and TEM analysis showed that most of the nanoparticles were spherical in shape. Zeta potential of AgNP was found to be - 33.5 mV which confirmed their high stability. FT-IR investigations confirmed the presence of different functional groups involved in the reduction and capping of AgNP. The synthesized AgNP showed effective DPPH (IC50-16.08 µg/mL), H2O2 (IC50-25.40 µg/mL), and superoxide (IC50-21.12 µg/mL) radical scavenging activities. These plant-AgNP showed significant inhibition of albumin denaturation (IC50-12.98 µg/mL) and antiproteinase activity (IC50-18.401 µg/mL). Besides, biosynthesized AgNP were found to have strong inhibitory effect against a cervical cancer (HeLa) cell line (IC50-5.418 µg/mL) as well as larvicidal activity against 3rd instar larvae of Anopheles stephensi (LC50-18.9 ppm, LC90-40.18 ppm), Aedes aegypti (LC50-12.395 ppm, LC90-36.34 ppm) and Culex quinquefasciatus (LC50-17.76 ppm, LC90-30.82 ppm) and were found to be non-toxic against normal cell line (HEK 293), and a non-target organism (Mesocyclops thermocyclopoides). This is the first report on the synthesis of AgNP using aqueous leaf extract of A. acuminata, validating their strong therapeutic potential.


Assuntos
Anti-Inflamatórios/farmacologia , Antineoplásicos/farmacologia , Antioxidantes/farmacologia , Atropa belladonna/química , Química Verde , Nanopartículas Metálicas/química , Extratos Vegetais/farmacologia , Prata/farmacologia , Animais , Culicidae , Flavonoides/análise , Sequestradores de Radicais Livres/farmacologia , Células HEK293 , Células HeLa , Humanos , Concentração de Íons de Hidrogênio , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Nanopartículas Metálicas/ultraestrutura , Fenóis/análise , Folhas de Planta/química , Nitrato de Prata/farmacologia , Espectrofotometria Ultravioleta , Taninos/análise , Temperatura , Fatores de Tempo , Difração de Raios X
16.
Toxicon ; 177: 52-88, 2020 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-32217234

RESUMO

Datura stramonium, Atropa belladonna, Hyoscyamus niger, and Scopolia carniolica are all temperate plants from the family Solanaceae, which as a result of their anticholinergic tropane alkaloids, hyoscyamine/atropine and scopolamine, have caused many cases of poisoning around the world. Despite the danger these nightshade plants represent, the literature often presents incomplete cases lacking in details and filled with ambiguity, and reviews on the topic tend to be limited in scope. Many also point to a gap in knowledge of these plants among physicians. To address this, the following review focuses on intoxications involving these plants as reported in the literature between 1966 and 2018, with brief mention to pertinent related plants to contextualise and provide a fuller picture of the situation surrounding the presently discussed temperate plants. Analysis of the literature displays that D. stramonium is largely associated with drug use among teens while A. belladonna is primarily ingested as a result of the berries being mistaken for edible fruits. H. niger was found to be largely ingested when mistaken for other plants, and S. carniolica was the cause of incredibly few intoxications.


Assuntos
Antagonistas Colinérgicos/toxicidade , Extratos Vegetais/toxicidade , Intoxicação por Plantas/epidemiologia , Solanaceae , Alcaloides de Solanáceas/toxicidade , Alcaloides , Atropa belladonna , Europa (Continente) , Humanos , Hyoscyamus , Escopolamina , Scopolia , Tropanos
17.
ACS Synth Biol ; 9(2): 437-448, 2020 02 21.
Artigo em Inglês | MEDLINE | ID: mdl-31935324

RESUMO

Ornithine decarboxylase (ODC) plays an important role in various biological processes; however, its role in plant secondary metabolism, especially in the biosynthesis of tropane alkaloids (TAs) such as pharmaceutical hyoscyamine, anisodamine, and scopolamine, remains largely unknown. In this study, we characterized the physiological and metabolic functions of the ODC gene of Atropa belladonna (AbODC) and determined its role in TA production using metabolic engineering approaches. Feeding assays with enzyme inhibitors indicated that ODC, rather than arginine decarboxylase (ADC), plays a major role in TA biosynthesis. Tissue-specific AbODC expression analysis and ß-glucuronidase (GUS) staining assays showed that AbODC was highly expressed in secondary roots, especially in the cylinder tissue. Enzymatic assays indicated that AbODC was able to convert ornithine to putrescine, with the highest activity at pH 8.0 and 30 °C. Additionally, AbODC showed higher catalytic efficiency than other plant ODCs, as evident from the Km, Vmax, and Kcat values of AbODC using ornithine as the substrate. In A. belladonna root cultures, suppression of AbODC greatly reduced the production of putrescine, N-methylputrescine, and TAs, whereas overexpression of AbODC significantly increased the biosynthesis of putrescine, N-methylputrescine, hyoscyamine, and anisodamine. Moreover, transgenic A. belladonna plants overexpressing AbODC showed a significantly higher production of hyoscyamine and anisodamine compared with control plants. These findings indicate that AbODC plays a key role in TA biosynthesis and therefore is a valuable candidate for increasing TA production in A. belladonna.


Assuntos
Atropa belladonna/enzimologia , Ornitina Descarboxilase/metabolismo , Tropanos/metabolismo , Alcaloides/metabolismo , Citosol/metabolismo , Concentração de Íons de Hidrogênio , Cinética , Engenharia Metabólica , Ornitina/metabolismo , Ornitina Descarboxilase/química , Ornitina Descarboxilase/genética , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Putrescina/biossíntese , Interferência de RNA , Alcaloides de Solanáceas/biossíntese
18.
Cell Mol Neurobiol ; 40(1): 179-188, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31456135

RESUMO

Recent advances in ethnobotanical and neurological research indicate that ingested plants from our diet may not only be a source of nutrition but also a source of biologically relevant nucleic-acid-encoded genetic information. A major source of RNA-encoded information from plants has been shown to be derived from small non-coding RNAs (sncRNAs) such as microRNAs (miRNAs) that can transfer information horizontally between plants and humans. In human hosts, the 3'-untranslated region (3'-UTR) of messenger RNAs (mRNAs) is targeted by these miRNAs to effectively down-regulate expression of that mRNA target in the host CNS. In this paper, we provide evidence that the Atropa belladonna aba-miRNA-9497 (miRBase conserved ID: bdi-miRNA-9497) is highly homologous to the CNS-abundant Homo sapiens miRNA-378 (hsa-miRNA-378) and both target the zinc-finger transcription factor ZNF-691 mRNA 3'-UTR to down-regulate ZNF-691 mRNA abundance. We speculate that the potent neurotoxic actions of the multiple tropane alkaloids of Atropa belladonna may be supplemented by the neuroregulatory actions of aba-miRNA-9497 on ZNF-691, and this may be followed by the modulation in the expression of ZNF-691-sensitive genes. This is the first example of a human brain-enriched transcription factor, ZNF-691, targeted and down-regulated by a naturally occurring plant microRNA, with potential to modulate gene expression in the human CNS and thus contribute to the neurotoxicological-and-psychoactive properties of the Atropa belladonna species of the deadly nightshade Solanaceae family.


Assuntos
Regiões 3' não Traduzidas/genética , Atropa belladonna/genética , MicroRNAs/metabolismo , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/genética , Dedos de Zinco/genética , Sequência de Bases , Humanos , MicroRNAs/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
19.
São Paulo; s.n; 2020. 29 p.
Tese em Português | HomeoIndex - Homeopatia, HomeoIndex - Homeopatia | ID: biblio-1122681

RESUMO

Relato de caso clínico de um paciente com Transtorno do Espectro Autista (TEA), através de anamnese e entrevista seguindo os critérios diagnósticos do DSM-V e o CID-10 6. Foram realizadas consultas, utilizando o Repertorio de Homeopatia5. Feita uma revisão bibliográfica, enfocando no tema do TEA e a homeopatia. Optou-se pela homeopatia como tratamento auxiliar no paciente autista do ambulatório-escola da Associação Paulista de Homeopatia.(AU)


Clinical case report of a patient with Autistic Spectrum Disorder (ASD), through anamnesis and interview following the diagnostic criteria of DSM-V and ICD-10. Consultations were carried out, using the Homeopathy Directory. A bibliographic review was made, focusing on the topic of ASD and homeopathy. Homeopathy was chosen as an auxiliary treatment for autistic patients at the outpatient school of the Associação Paulista de Homeopatia.(AU)


Assuntos
Humanos , Masculino , Criança , Atropa belladonna , Transtorno do Espectro Autista/terapia , Homeopatia
20.
ACS Synth Biol ; 8(3): 474-481, 2019 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-30721031

RESUMO

Enzymes are the ultimate entities responsible for chemical transformations in natural and engineered biosynthetic pathways. However, many natural enzymes suffer from suboptimal functional expression due to poor intrinsic protein stability. Further, stability enhancing mutations often come at the cost of impaired function. Here we demonstrate an automated protein engineering strategy for stabilizing enzymes while retaining catalytic function using deep mutational scanning coupled to multiple-filter based screening and combinatorial mutagenesis. We validated this strategy by improving the functional expression of a Type III polyketide synthase from the Atropa belladonna biosynthetic pathway for tropane alkaloids. The best variant had a total of 8 mutations with over 25-fold improved activity over wild-type in E. coli cell lysates, an improved melting temperature of 11.5 ± 0.6 °C, and only minimal reduction in catalytic efficiency. We show that the multiple-filter approach maintains acceptable sensitivity with homology modeling structures up to 4 Å RMS. Our results highlight an automated protein engineering tool for improving the stability and solubility of difficult to express enzymes, which has impact for biotechnological applications.


Assuntos
Aciltransferases/química , Aciltransferases/genética , Atropa belladonna/enzimologia , Biotecnologia/métodos , Ciência de Dados/métodos , Engenharia de Proteínas/métodos , Aciltransferases/metabolismo , Alcaloides de Belladona/metabolismo , Vias Biossintéticas , Códon sem Sentido , Estabilidade Enzimática/genética , Escherichia coli/metabolismo , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Substâncias Luminescentes/química , Substâncias Luminescentes/metabolismo , Mutagênese , Mutação de Sentido Incorreto , Saccharomyces cerevisiae/metabolismo , Solubilidade , Temperatura de Transição
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