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Cardiovascular & Hematological... 2020The relationship between vascular damage and diabetes mellitus was exploited using avocado seed extracts. The purpose of the study was to understand the therapeutic...
BACKGROUND AND OBJECTIVES
The relationship between vascular damage and diabetes mellitus was exploited using avocado seed extracts. The purpose of the study was to understand the therapeutic relevance of glycosides compared to standard vascular and anti-diabetic drugs. Constituent Avocado Seed Glycosides (ASG) were analysed and administered to rats with Diabetes-Induced Vascular Damage (DIVD).
METHODS
The rats were first administered with streptozotocin and screened after seven days for alterations in blood glucose, insulin, vascular cell adhesion molecule (VCAM-1), Von Willebrand factor (VWF), Renin-Angiotensin-Aldosterone System (RAS), eNOx, and endothelin-1 (ET-1). Only rats that satisfied these criteria were recruited and treated with either glibenclamide, met.su + losart, or 200 mg/kg body weight ASG for 28 days.
RESULTS
There was an abundance of digitoxin (13.41 mg/100g), digoxin (17.98 mg/100g), avicularin (165.85 mg/100g), and hyperoside (282.51 mg/100g). ASG or met.su + losart exhibited slight modulatory properties on glucose homeostasis. Rats with DIVD showed elevated renin, angiotensin, VCAM-1 and Lp-PLA2 levels but slightly decreased with glibenclamide treatment and normalized with ASG or met.su + losart administration. All treatments normalized Hcy levels. DIVD caused the overproduction of CnT, LDH, Crt-K, LDL-c, TG, and TC and suppressed HDL-c but was completely normalized by the ASG. Water intake remained altered in treated rats.
CONCLUSION
The ASG had no relevant effect on glucose homeostasis during DIVD but showed significant vasoprotective properties.
Topics: Animals; Diabetes Mellitus, Experimental; Endothelium, Vascular; Glycosides; Hypoglycemic Agents; Persea; Plant Extracts; Protective Agents; Rats; Seeds
PubMed: 32386502
DOI: 10.2174/1871529X20666200510012012 -
Data in Brief Jun 2020Cardiac glycosides, steroid derivatives extracted from the foxglove plants, have been used for the treatment of heart failure since the 18th century. A method based on...
Cardiac glycosides, steroid derivatives extracted from the foxglove plants, have been used for the treatment of heart failure since the 18th century. A method based on liquid chromatography coupled with high-resolution tandem mass spectrometry (LC/MS) has been developed to characterize and quantify cardiac glycosides in fresh-leaf extracts of the foxglove () plants [1]. In this report, the fragmentation spectra of additional authentic standards of cardiac glycoside (digitoxigenin, digoxigenin, -acetyldigoxin) and cardenolides identified in the leaves of () and () were provided with high resolution. The exact mass of signature peaks for the aglycones and the sugar units of cardenolides were measured. This dataset is valuable to researchers interested in characterizing cardenolides in plants, or quantifying cardenolides in drug tablets, or studying cardenolide toxicities in animals. The fragmentation patterns of authentic cardenolide standards provided in these data can be used to validate relevant cardenolides in various biological samples and to infer chemical structures of unknown cardiac glycosides.
PubMed: 32300626
DOI: 10.1016/j.dib.2020.105464 -
Toxins Apr 2020Cardiac glycosides (CGs) are naturally occurring plant secondary metabolites that can be toxic to humans and animals. The aim of this work was to develop a targeted...
Development and Validation of a UHPLC-ESI-MS/MS Method for Quantification of Oleandrin and Other Cardiac Glycosides and Evaluation of Their Levels in Herbs and Spices from the Belgian Market.
Cardiac glycosides (CGs) are naturally occurring plant secondary metabolites that can be toxic to humans and animals. The aim of this work was to develop a targeted analytical method utilizing liquid chromatography-tandem mass spectrometry (LC-MS/MS) for quantification of these plant toxins in a herbal-based food and human urine. The method included oleandrin, digoxin, digitoxin, convallatoxin, and ouabain. Samples of culinary herbs were extracted with acetonitrile and cleaned using Oasis MAX solid-phase extraction (SPE), while samples of urine were diluted with acidified water and purified on Oasis HLB SPE cartridges. Limits of quantification were in the range of 1.5-15 ng/g for herbs and 0.025-1 ng/mL for urine. The mean recovery of the method complied with the acceptable range of 70-120% for most CGs, and relative standard deviations were at maximum 14% and 19% for repeatability and reproducibility, respectively. Method linearity was good with calculated R² values above 0.997. The expanded measurement uncertainty was estimated to be in the range of 7-37%. The LC-MS/MS method was used to examine 65 samples of culinary herbs and herb and spice mixtures collected in Belgium, from supermarkets and local stores. The samples were found to be free from the analyzed CGs.
Topics: Belgium; Cardenolides; Cardiac Glycosides; Chromatography, High Pressure Liquid; Humans; Limit of Detection; Plant Preparations; Reproducibility of Results; Spectrometry, Mass, Electrospray Ionization; Spices; Supermarkets; Tandem Mass Spectrometry; Urinalysis
PubMed: 32283845
DOI: 10.3390/toxins12040243 -
Archives of Pathology & Laboratory... Sep 2020Immunoassays using the interaction between streptavidin and biotin are used for clinical chemical analytes on platforms by many different manufacturers. The design can...
CONTEXT.—
Immunoassays using the interaction between streptavidin and biotin are used for clinical chemical analytes on platforms by many different manufacturers. The design can be susceptible to interference from high-dose biotin intake in patients, which remains an often-overlooked confounder despite recently increased awareness.
OBJECTIVE.—
To evaluate an easily implementable method of in vitro biotin depletion for the removal of biotin interference in immunoassays for potentially time-critical analytes.
DESIGN.—
A biotin stock solution was made and de-identified patient samples were spiked to reach a biotin concentration of 1.126 × 106 pg/mL, the maximum reported biotin concentration 1 to 2 hours after a single oral dose of 300 mg biotin. Then, the resulting interference in Elecsys immunoassays for cortisol, cyclosporine A, tacrolimus, digitoxin, thyroid-stimulating hormone, free triiodothyronine, free thyroxine, C-peptide, insulin, N-terminal pro-B-type natriuretic peptide, troponin T high sensitive, human immunodeficiency virus, procalcitonin, β human chorionic gonadotropin, toxoplasma immunoglobulin M, and toxoplasma immunoglobulin G was evaluated before and after biotin depletion using streptavidin particles.
RESULTS.—
All tested immunoassays, with the exception of toxoplasma immunoglobulin M and toxoplasma immunoglobulin G, suffered from significant biotin interference. The depletion protocol removed assay interference due to biotin and produced results that were close or identical to initial prespike measurements.
CONCLUSIONS.—
Despite an increase in turnaround times, biotin adsorption is a feasible countermeasure for biotin interference in Elecsys immunoassays. Until test kits with an increased resistance to the interference from high-dose biotin intake are distributed, the evaluated protocol can provide results properly reflecting the patient's clinical condition.
Topics: Biotin; Humans; Immunoassay
PubMed: 31944861
DOI: 10.5858/arpa.2019-0425-OA -
Biochimica Et Biophysica Acta. General... Apr 2020Imatinib mesylate (imatinib) is the first-line treatment for newly diagnosed chronic myeloid leukemia (CML) due to its remarkable hematologic and cytogenetic responses....
BACKGROUND
Imatinib mesylate (imatinib) is the first-line treatment for newly diagnosed chronic myeloid leukemia (CML) due to its remarkable hematologic and cytogenetic responses. We previously demonstrated that the imatinib-resistant CML cells (Myl-R) contained elevated Lyn activity and intracellular creatine pools compared to imatinib-sensitive Myl cells.
METHODS
Stable isotope metabolic labeling, media creatine depletion, and Na/K-ATPase inhibitor experiments were performed to investigate the origin of creatine pools in Myl-R cells. Inhibition and shRNA knockdown were performed to investigate the specific role of Lyn in regulating the Na/K-ATPase and creatine uptake.
RESULTS
Inhibition of the Na/K-ATPase pump (ouabain, digitoxin), depletion of extracellular creatine or inhibition of Lyn kinase (ponatinib, dasatinib), demonstrated that enhanced creatine accumulation in Myl-R cells was dependent on uptake from the growth media. Creatine uptake was independent of the Na/creatine symporter (SLC6A8) expression or de novo synthesis. Western blot analyses showed that phosphorylation of the Na/K-ATPase on Tyr 10 (Y10), a known regulatory phosphorylation site, correlated with Lyn activity. Overexpression of Lyn in HEK293 cells increased Y10 phosphorylation (pY10) of the Na/K-ATPase, whereas Lyn inhibition or shRNA knockdown reduced Na/K-ATPase pY10 and decreased creatine accumulation in Myl-R cells. Consistent with enhanced uptake in Myl-R cells, cyclocreatine (Ccr), a cytotoxic creatine analog, caused significant loss of viability in Myl-R compared to Myl cells.
CONCLUSIONS
These data suggest that Lyn can affect creatine uptake through Lyn-dependent phosphorylation and regulation of the Na/K-ATPase pump activity.
GENERAL SIGNIFICANCE
These studies identify kinase regulation of the Na/K-ATPase as pivotal in regulating creatine uptake and energy metabolism in cells.
Topics: Antineoplastic Agents; Cell Proliferation; Cell Survival; Creatine; Drug Resistance, Neoplasm; Drug Screening Assays, Antitumor; Humans; Imatinib Mesylate; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; Tumor Cells, Cultured; src-Family Kinases
PubMed: 31881245
DOI: 10.1016/j.bbagen.2019.129507 -
Case Reports in Cardiology 2019Foxglove ( L.) leaves are frequently confused with borage ( L.), which is traditionally used as a food ingredient. Due to the presence of the cardiac glycosides, mostly...
Foxglove ( L.) leaves are frequently confused with borage ( L.), which is traditionally used as a food ingredient. Due to the presence of the cardiac glycosides, mostly digitoxin, foxglove leaves are poisonous to human and may be fatal if ingested. A 55-year-old Caucasian woman complaining weakness, fatigue, nausea, and vomiting was admitted to the Emergency Department. Her symptoms started following consumption of a home-made savory pie with 5 leaves from a plant bought in a garden nursery as borage. Digoxinemia was high (10.4 g/L). The patient was admitted to the cardiac intensive care unit for electrocardiographic monitoring. Two days after admission, a single episode of advanced atrioventricular (AV) block was recorded by telemetry, followed by a second-degree AV block episode. Plasma samples at day 11 were analysed by LC-MS spectrometry, and gitoxin was identified suggesting that this compound may be responsible for the clinical toxicity rather than digoxin. In the case of spp. poisoning, laboratory data should be interpreted according to the clinical picture and method of analysis used since a variety of glycosides, which are chemically similar to the cardioactive glycosides but without or with fewer cardiac effects, may be incorrectly recognized as digoxin by the test, giving misleading results.
PubMed: 31871798
DOI: 10.1155/2019/9707428 -
Respiratory Research Dec 2019Several small molecule corrector and potentiator drugs have recently been licensed for Cystic Fibrosis (CF) therapy. However, other aspects of the disease, especially...
BACKGROUND
Several small molecule corrector and potentiator drugs have recently been licensed for Cystic Fibrosis (CF) therapy. However, other aspects of the disease, especially inflammation, are less effectively treated by these drugs. We hypothesized that small molecule drugs could function either alone or as an adjuvant to licensed therapies to treat these aspects of the disease, perhaps emulating the effects of gene therapy in CF cells. The cardiac glycoside digitoxin, which has been shown to inhibit TNFα/NFκB signaling in CF lung epithelial cells, may serve as such a therapy.
METHODS
IB3-1 CF lung epithelial cells were treated with different Vertex (VX) drugs, digitoxin, and various drug mixtures, and ELISA assays were used to assess suppression of baseline and TNFα-activated secretion of cytokines and chemokines. Transcriptional responses to these drugs were assessed by RNA-seq and compared with gene expression in AAV-[wildtype]CFTR-treated IB3-1 (S9) cells. We also compared in vitro gene expression signatures with in vivo data from biopsied nasal epithelial cells from digitoxin-treated CF patients.
RESULTS
CF cells exposed to digitoxin exhibited significant suppression of both TNFα/NFκB signaling and downstream secretion of IL-8, IL-6 and GM-CSF, with or without co-treatment with VX drugs. No evidence of drug-drug interference was observed. RNA-seq analysis showed that gene therapy-treated CF lung cells induced changes in 3134 genes. Among these, 32.6% were altered by digitoxin treatment in the same direction. Shared functional gene ontology themes for genes suppressed by both digitoxin and gene therapy included inflammation (84 gene signature), and cell-cell interactions and fibrosis (49 gene signature), while genes elevated by both were enriched for epithelial differentiation (82 gene signature). A new analysis of mRNA data from digitoxin-treated CF patients showed consistent trends in expression for genes in these signatures.
CONCLUSIONS
Adjuvant gene therapy-emulating activities of digitoxin may contribute to enhancing the efficacy of currently licensed correctors and potentiators in CF patients.
Topics: Animals; Cardiotonic Agents; Cells, Cultured; Cystic Fibrosis; Digitoxin; Dose-Response Relationship, Drug; Genetic Therapy; Humans; Inflammation Mediators; Rats; Rats, Inbred F344; Respiratory Mucosa; Treatment Outcome
PubMed: 31864360
DOI: 10.1186/s12931-019-1214-8 -
Food Chemistry Feb 2020Protein conformation and the 3D water structure play important roles in the ability of bovine serum albumin (BSA) to form stable nanostructures with bioactive molecules....
Protein conformation and the 3D water structure play important roles in the ability of bovine serum albumin (BSA) to form stable nanostructures with bioactive molecules. We studied the influence of BSA unfolding and those of two Hofmeister salts, sodium chloride (NaCl) as kosmotrope and sodium thiocyanate (NaSCN) as chaotrope, on BSA/lutein binding at pH 7.4 using fluorescence spectroscopy. The BSA/lutein complex formation was entropically driven and lutein was preferentially bound to site III of BSA. The binding constant (10 L mol), complex stoichiometry (1:1), and thermodynamic potential for BSA/lutein binding were independent of protein conformation and Hofmeister salts. However, the enthalpic and entropic components of BSA/lutein binding in the presence of NaSCN decreased as the temperature increased. The opposite was observed for BSA/lutein binding in the presence of NaCl and for denatured BSA/lutein binding. Therefore, the BSA conformation and 3D water structure directly affected the BSA/lutein binding thermodynamics.
Topics: Animals; Binding Sites; Cattle; Lutein; Protein Binding; Protein Conformation; Salts; Serum Albumin, Bovine; Sodium Chloride; Spectrometry, Fluorescence; Temperature; Thermodynamics; Thiocyanates
PubMed: 31520921
DOI: 10.1016/j.foodchem.2019.125463 -
Shokuhin Eiseigaku Zasshi. Journal of... 2019Assuming the intentional adulteration of beverages with plant toxins, an LC-MS/MS method for the simultaneous determination of 18 plant toxins (lycorine, galantamine,...
Assuming the intentional adulteration of beverages with plant toxins, an LC-MS/MS method for the simultaneous determination of 18 plant toxins (lycorine, galantamine, ricinine, scopolamine, gelsemine, atropine, colchicine, α-solanine, jervine, α-chaconine, veratramine, mesaconitine, digoxin, protoveratrine A, aconitine, hypaconitine, oleandrin, and digitoxin) was developed. As analytical samples, beer, distilled spirits, blend tea, ready-to-drink (RTD) coffee, and fermented milk drink were selected. The extraction and purification of the analytes were performed using modified QuEChERS method. Method validation in terms of intra-day precision, accuracy, and extraction recovery obtained satisfactory results. The calibration curves for the analytes were linear from 5 to 200 ng/mL (r>0.990), which enabled the determination of toxins in even trace amounts.
Topics: Beverages; Chromatography, Liquid; Food Analysis; Tandem Mass Spectrometry; Toxins, Biological
PubMed: 31474651
DOI: 10.3358/shokueishi.60.108 -
Frontiers in Oncology 2019Castration Resistant Prostate Cancer (CRPC) is thought to be driven by a collaborative mechanism between TNFα/NFκB and TGFβ signaling, leading to inflammation,...
Castration Resistant Prostate Cancer (CRPC) is thought to be driven by a collaborative mechanism between TNFα/NFκB and TGFβ signaling, leading to inflammation, Epithelial-to-Mesenchymal-Transition (EMT), and metastasis. Initially, TGFβ is a tumor suppressor, but in advanced metastatic disease it switches to being a tumor promoter. TGFBR2 may play a critical role in this collaboration, as its expression is driven by NFκB and it is the primary receptor for TGFβ. We have previously reported that the cardenolide drug digitoxin blocks TNFα/NFκB-driven proinflammatory signaling. We therefore hypothesized that digitoxin might break the collaborative process between NFκB and TGFβ by also inhibiting expression of TGFBR2. We therefore tested whether TGFβ-driven EMT and resulting metastases would be suppressed. Here we show, , that digitoxin inhibits NFκB-driven TGFBR2 expression, as well as Vimentin, while elevating E-cadherin expression. Digitoxin also significantly reduces HSPB1 mRNA and the HSPB1/RBFOX2 mRNA ratio in PC3 cells. , in a syngeneic, immune competent rat model of metastatic CRPC, we show that digitoxin also suppresses expression, as well as expression of other genes classically driven by NFκB, and of multiple EMT genes associated with metastasis. Concurrently, digitoxin suppresses tumor growth and metastasis in these animals, and prolongs survival. Gross tumor recurrence following tumor resection also appears prevented in 30% of cases. While the existence of a collaboration between NFκB and TGFβ to drive EMT and metastasis has previously been appreciated, we show here, for the first time, that chronic, low concentrations of digitoxin are able to block CRPC tumor progression, EMT and the ensuing metastatic disease.
PubMed: 31428571
DOI: 10.3389/fonc.2019.00630