Cardiac Steroids Research Articles

Thevetia thevetioides Cardenolide and Related Cardiac Glycoside Profile in Mature and Immature Seeds by High-Resolution Thin-Layer Chromatography (HPTLC) and Quadrupole Time of Flight-Tandem Mass Spectrometry (Q-TOF MS/MS) Reveals Insights of the Cardenolide Biosynthetic Pathway.

Thevetia thevetioides is a species within the Apocynaceae family known for containing cardenolide-glycosides, commonly referred to as cardiac glycosides, which are characteristic of this genus. The seeds of the Thevetia species are frequently used as a model source for studying cardiac steroids, as these glycosides can be more readily extracted from the oil-rich seeds than from the plant's green tissues. In this work, the cardenolide profile of ripe and immature seeds was determined and compared to establish the main differences. Ripe seeds contain six related cardenolides and triosides, with thevetin B being the predominant component. In contrast, immature seeds exhibit a total of thirteen cardiac glycosides, including monoglycosides such as neriifolin and peruvosides A, B, and C, as well as diglycosides like thevebiosides A, B, and C. Some of these compounds have previously been identified as degradation products of more complex cardiac glycosides; however, their presence in immature seeds, as described in this study, suggests that they may serve as biosynthetic precursors to the triosides observed in mature seeds. The glycoside patterns observed via HPTLC are associated with specific chemical structures characteristic of this genus, typically featuring thevetose or acetyl-thevetose at the first position, followed by glucose or gentibiose in di- or trisaccharides, independent of the trioside aglycones identified: digitoxigenin, cannogenin, or yccotligenin. Ripe seeds predominantly contain triosides, including thevetin B, C, and A, the latter of which has not been previously reported.

Involvement of the Na+, K+-ATPase α1 Isoform and Endogenous Cardiac Steroids in Depression- and Manic-like Behaviors.

Bipolar disorder (BD) is a severe and common chronic mental illness characterized by recurrent mood swings between depression and mania. The biological basis of the disease is poorly understood, and its treatment is unsatisfactory. Na+, K+-ATPase is a major plasma membrane transporter and signal transducer. The catalytic α subunit of this enzyme is the binding site for cardiac steroids. Three α isoforms of the Na+, K+-ATPase are present in the brain. Previous studies have supported the involvement of the Na+, K+-ATPase and endogenous cardiac steroids (ECS) in the etiology of BD. Decreased brain ECS has been found to elicit anti-manic and anti-depressive-like behaviors in mice and rats. However, the identity of the specific α isoform involved in these behavioral effects is unknown. Here, we demonstrated that decreasing ECS through intracerebroventricular (i.c.v.) administration of anti-ouabain antibodies (anti-Ou-Ab) decreased the activity of α1+/- mice in forced swimming tests but did not change the activity in wild type (wt) mice. This treatment also affected exploratory and anxiety behaviors in α1+/- but not wt mice, as measured in open field tests. The i.c.v. administration of anti-Ou-Ab decreased brain ECS and increased brain Na+, K+-ATPase activity in wt and α1+/- mice. The serum ECS was lower in α1+/- than wt mice. In addition, a study in human participants demonstrated that serum ECS significantly decreased after treatment. These results suggest that the Na+, K+-ATPase α1 isoform is involved in depressive- and manic-like behaviors and support that the Na+, K+-ATPase/ECS system participates in the etiology of BD.

Strategies for senolytic drug discovery.

Senolytics are a category of drugs that reduce the impact of cellular senescence, an effect associated with a range of chronic and age-related diseases. Since the discovery of the first senolytics in 2015, the number of known senolytic agents has grown dramatically. This review discusses the broad categories of known senolytics-kinase inhibitors, Bcl-2 family protein inhibitors, naturally occurring polyphenols, heat shock protein inhibitors, BET family protein inhibitors, P53 stabilizers, repurposed anti-cancer drugs, cardiac steroids, PPAR-alpha agonists, and antibiotics. The approaches used to screen for new senolytics are articulated including a range of methods to induce senescence, different target cell types, various senolytic assays, and markers. The choice of methods can greatly influence the outcomes of a screen, with high-quality screens featuring robust systems, adequate controls, and extensive validation in alternate assays. Recent advances in single-cell analysis and computational methods for senolytic identification are also discussed. There is significant potential for further drug discovery, but this will require additional research into drug targets and mechanisms of actions and their subsequent rigorous evaluation in pre-clinical models and human trials.

Beneficial effect of digoxin-specific Fab fragments in bipolar disorder- a preliminary study

A vast amount of experimental data supports the notion that Na+, K+-ATPase and endogenous cardiac steroids (ECS) such as digoxin and ouabain are involved in bipolar disorders. The aim of the study was to examine the possible beneficial effect of Digoxin-specific antibody Fab fragments Digibind, in bipolar disorder (BD) patients. In a pilot, open-label study without a control group, BD inpatient subjects received a single dose of Digibind, injected intravenously. During the 72 hrs following the intervention. the response to Digibind was measured using the Montgomery-Asberg Depression Rating Scale (MADRS). A total 6 patients with a moderate to severe bipolar depressive episode (MADRS 30.3, 24–38) participated in the study. We found a significant clinical improvement at all time points after Digibind administration, manifested by a maximal decrease in the MADRS score of 8.3 points at 24 hrs, followed by a gradual rebound to baseline scores. No control group was included, treatment was open-label, and only 6 patients were studied. This preliminary study showed that a one-time injection of Digibind resulted in a transient significant beneficial effect on depression symptoms in BD patients.

Automated machine learning approach for developing a quantitative structure-activity relationship model for cardiac steroid inhibition of Na+/K+-ATPase.

Quantitative structure-activity relationship (QSAR) modeling is a method of characterizing the relationship between chemical structures and biological activity. Automated machine learning enables computers to learn from large datasets and can be used for chemoinformatics. Cardiac steroids (CSs) inhibit the activity of Na+/K+-ATPase (NKA) in several species, including humans, since the binding pocket in which NKA binds to CSs is highly conserved. CSs are used to treat heart disease and have been developed into anticancer drugs for use in clinical trials. Novel CSs are, therefore, frequently synthesized and their activities evaluated. The purpose of this study is to develop a QSAR model via automated machine learning to predict the potential inhibitory activity of compounds without performing experiments. The chemical structures and inhibitory activities of 215 CS derivatives were obtained from the scientific literature. Predictive QSAR models were constructed using molecular descriptors, fingerprints, and biological activities. The best predictive QSAR models were selected based on the LogLoss value. Using these models, the Matthews correlation coefficient, F1 score, and area under the curve of the test dataset were 0.6729, 0.8813, and 0.8812, respectively. Next, we showed automated construction of the predictive models for CS derivatives, which may be useful for identifying novel CSs suitable for candidate drug development. The automated machine learning-based QSAR method developed here should be applicable for the time-efficient construction of predictive models using only a small number of compounds.

Synthesis and Biological Evaluation of Novel Bufalin Derivatives.

Bufalin and other cardiac steroids (CS) have been used for centuries for the treatment of congestive heart failure, arrhythmias, and other maladies. However, toxicity and the small therapeutic window of this family of steroids limit their use. Therefore, attempts to synthesize a potent, but less toxic, CS are of major importance. In the present study, two novel bufalin derivatives were synthesized and some of their pharmacological properties were characterized. The reaction of bufalin with Ishikawa’s reagent resulted in the production of two novel bufalin derivatives: bufalin 2,3-ene and bufalin 3,4-ene. The compounds were purified with TLC and HPLC and their structure was verified with UV, NMR, and MS analyses. The biological activities of these compounds were evaluated by testing their ability to inhibit the Na+, K+-ATPase activity of the brain microsomal fraction to induce cytotoxic activity against the NCI-60 human tumor cell line panel and non-cancer human cells, and to increase the force of contraction of quail embryonic heart muscle cells in culture. The two steroids exhibited biological activities similar to those of other CS in the tested experimental systems, but with reduced cytotoxicity, advocating their development as drugs for the treatment of heart failure and arrhythmias.

Endogenous Cardiac Steroids in Bipolar Disorder: State of the Art.

Bipolar disorder (BD) is a severe psychiatric illness with a poor prognosis and problematic, suboptimal, treatments. Treatments, borne of an understanding of the pathoetiologic mechanisms, need to be developed in order to improve outcomes. Dysregulation of cationic homeostasis is the most reproducible aspect of BD pathophysiology. Correction of ionic balance is the universal mechanism of action of all mood stabilizing medications. Endogenous sodium pump modulators (collectively known as endogenous cardiac steroids, ECS) are steroids which are synthesized in and released from the adrenal gland and brain. These compounds, by activating or inhibiting Na+, K+-ATPase activity and activating intracellular signaling cascades, have numerous effects on cell survival, vascular tone homeostasis, inflammation, and neuronal activity. For the past twenty years we have addressed the hypothesis that the Na+, K+-ATPase-ECS system may be involved in the etiology of BD. This is a focused review that presents a comprehensive model pertaining to the role of ECS in the etiology of BD. We propose that alterations in ECS metabolism in the brain cause numerous biochemical changes that underlie brain dysfunction and mood symptoms. This is based on both animal models and translational human results. There are data that demonstrate that excess ECS induce abnormal mood and activity in animals, while a specific removal of ECS with antibodies normalizes mood. There are also data indicating that circulating levels of ECS are lower in manic individuals, and that patients with BD are unable to upregulate synthesis of ECS under conditions that increase their elaboration in non-psychiatric controls. There is strong evidence for the involvement of ion dysregulation and ECS function in bipolar illness. Additional research is required to fully characterize these abnormalities and define future clinical directions.

Neuroinflammation and Neutrophils: Modulation by Ouabain.

Cardiotonic steroids are natural compounds that present many physiological and pharmacological functions. They bind Na+/K+-ATPase (NKA) modifying cellular ion concentration and trigger cell signaling mechanisms without altering ion balance. These steroids are known to modulate some immune responses, including cytokine production, neutrophil migration, and inflammation (peripherally and in the nervous system). Inflammation can occur in response to homeostasis perturbations and is related to the development of many diseases, including immune-mediated diseases and neurodegenerative disorders. Considering the neutrophils role in the general neuroinflammatory response and that these cells can be modulated by cardiac steroids, this work aims to review the possible regulation of neutrophilic neuroinflammation by the cardiac steroid ouabain.

Quality Assessment of Selected Anti-Asthmatic Herbal Products Marketed in Kaduna State Metropolis, Nigeria

Background: Herbal medicines are the oldest form of healthcare known to humanity and it is used worldwide for prevention and treatment of different disease conditions. The presence and proliferation of substandard herbal medicines in the market that do not fulfil standards set by the regulating bodies in Nigerian is a matter of concern. Objective: To assess quality of some anti-asthmatic herbal products marketed in Kaduna State using standard methods. Method: Ten (10) different samples of anti-asthmatic herbal products marketed in Kaduna metropolis were selected randomly and standard methods by World Health Organization methods, Trease and Evans, and digestion by AOAC were used to assess the quality of the herbal samples. Results: The result obtained showed that 50 % of the herbal preparations contain moisture content higher than the permissible limit (8 %), while 40 and 10 % of the samples had water extractive and alcohol extractive values respectively lower than the 15 % minimum limit stipulated by European Pharmacopeia. Also, 30 % of the samples had higher ash values than the 14 % maximum acceptable limit recommended by European Pharmacopoeia. All the herbal preparations analyzed contain cardiac glycosides, carbohydrates, alkaloids, steroids and flavonoid, while 80 % of the samples contain terpenoids, 70 % of the samples contain saponins and 50 % contain tannins. Also, 80 and 50 % of the samples analyzed contained cadmium and lead respectively above WHO permissible limits (0.3 and 10 ppm respectively). Conclusion: None of the anti-asthmatic herbal products analyzed passed the quality assessment carried out in this study.

Microwave assisted extraction, phytochemical and antimicrobial evaluation of ethyl acetate extracts of stem bark of Ficus exasperata (Vahl)

Stem bark of Ficus exasperata was extracted using ethyl acetate by microwave-assisted solvent extraction. Phytochemical screening and antimicrobial evaluation were carried out on the extract. Phytochemical screening revealed the presence of cardiac glycosides, tannins, flavonoids, triterpenes and steroids. Antimicrobial evaluation revealed that the extract is active against Methicillin – resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus pneumoniae, Salmonella typhi, Proteus mirabilis, Shigella dysenteriae, Candida stellatoidea and Candida tropicalis. The zones of inhibition (mm) for the test organisms were (24 - 31) mm. Minimum inhibition concentrations (mg/L) of extract against MRSA, Staphylococcus aureus, Streptococcus pneumonia, Salmonella typhi, Proteus mirabilis, Shigella dysenteriae, Candida stellatoidea and Candida tropicalis were 2.5, 1.25,1.25, 2.5, 1.25, 1.25, 1.25 and 2.5 respectively. Minimum bactericidal concentrations (mg/L) of extract against Methicillin – resistant Staphylococcus aureus (MRSA), Staphylococcus aureus, Streptococcus pneumonia, Salmonella typhi, Proteus mirabilis Shigella dysenteriae Candida stellatoidea and Candida tropicalis were 5, 2.5, 2.5, 5, 5, 2.5,2.5 and 5 respectively. The extract showed high inhibition against MRSA and Candida tropicalis.

Preliminary sub-acute toxicological assessment of methanol leaves extract of Culcasia angolensis (Araceae) in Wistar rats

BackgroundThe plant Culcasia angolensis (Araceae) has diverse ethnomedicinal uses, including the management of rheumatic pain, healing of cuts, dislocations, and bruises. Despite its potential therapeutic uses, the toxicity profile of Culcasia angolensis has not been evaluated. This study assessed the sub-acute toxicity effects of Culcasia angolensis leaves extract (CAE). The phytochemical determination of the CAE was conducted as per the standard protocols. The median lethal dose (LD50) was determined using the Organization for Economic Cooperation and Development (OECD) 423 guideline. Besides, the sub-acute toxic effects of the CAE (125, 250, and 500 mg/kg) were investigated following administration of the CAE daily for 28 consecutive days as per the OECD 407 guideline. The weekly body weights were recorded. The animals were euthanized on the 29th day, and blood samples were obtained for haematological and biochemical investigations. The heart, kidney, liver, and lungs were collected for histological examinations. Besides, the relative organ weights (ROW) were determined.ResultsThe CAE contains cardiac glycosides, alkaloids, tannins, flavonoids, steroids, saponins, and terpenoids. The oral LD50 was above 5 g/kg. There was a remarkable decline in the weekly body weight at all the CAE doses. The CAE increased the lymphocytes, aspartate transaminase, and urea. However, the levels of alanine transaminase and alkaline phosphatase were elevated remarkably. The histological studies did not reveal any serious organs abnormalities.ConclusionThe CAE is relatively safe on acute administration. However, it may be slightly toxic on sub-acute administration, especially to the liver and kidney.

Ouabain inhibitor rostafuroxin attenuates dextromethorphan-induced manic potential

Dextromethorphan (DM) abuse produces mania-like symptoms in humans. ERK/Akt signaling activation involved in manic potential can be attenuated by the inhibition of ouabain-like cardiac steroids. In this study, increased phosphorylations of ERK/Akt and hyperlocomotion induced by DM (30 mg/kg, i.p./day × 7) were significantly protected by the ouabain inhibitor rostafuroxin (ROSTA), suggesting that DM induces the manic potential. ROSTA significantly attenuated DM-induced protein kinase C δ (PKCδ) phosphorylation, GluN2B (i.e., MDA receptor subunit) expression, and phospho-PKCδ/GluN2B interaction. DM instantly upregulated the nuclear factor erythroid-2-related factor 2 (Nrf2)-dependent system. However, DM reduced Nrf2 nuclear translocation, Nrf2 DNA binding activity, γ-glutamylcysteine mRNA expression, and subsequent GSH/GSSG level and enhanced oxidative parameters following 1-h of administration. ROSTA, PKCδ inhibitor rottlerin, and GluN2B inhibitor traxoprodil significantly attenuated DM-induced alterations in Nrf2-related redox parameters and locomotor activity induced by DM in wild-type mice. Importantly, in PKCδ knockout mice, DM failed to alter the above parameters. Further, ROSTA and traxoprodil also failed to enhance PKCδ depletion effect, suggesting that PKCδ is a critical target for the anti-manic potential of ROSTA or GluN2B antagonism. Our results suggest that ROSTA inhibits DM-induced manic potential by attenuating ERK/Akt activation, GluN2B/PKCδ signalings, and Nrf2-dependent system.

A Review on: Digitalis Purpurea

Digitalis contain cardiac glycosides and used in the treatment of congestive heart failure. Digitalis drug obtain from Digitalis Purpurea L. belongs to family scrophulariaceae. 200 years ago William Withering discover the use of digitalis in CHF treatment in 1785.In 18th century a controversy has existed about the benefits and risks of digitalis in patients with heart problems. Active component of digitalis classified as cardiac glycosides and cardiotonic steroids. The term "digitalis" or "cardiac glycosides" are used to refer to any of steroids or steroid glycosides compound which exert characteristic positively inotropic and electrophysiological effect on heart. In 20th century digitalis and its derivatives specially, digoxin is used in treatment of CHF. Cardiac glycosides are used in therapy of CHF since William Withering identify their uses. Digitalis purpurea is not only use in treatment of CHF also has wound healing, antioxidant and cytotoxic activity. In 1990 most prescribed drug is digoxin. It shows intermediate duration of action. Digoxin is use for its rate control in atrial fibrillation in the presence of marginal blood pressure. The introduction of diuretic provide an alternative more effective and safer for treatment of CHF.
 Keywords: CHF., Digitalis Purpurea L

Antimicrobial, antioxidant and quantitative phytochemical evaluation of the dichloromethane and methanolic leaf extracts of Solenostemon monostychus P. Beauv

Solenostemon monostachyus P. Beauv (Lamiaceae family) is an important herb that is widespread in West and Central Africa.The leaves are used in various decoctions traditionally to treat diseases most especially microbial infections associated with the foot. In this study, the phytochemicals (qualitative and quantitative) of the plant were determined. The antimicrobial and antioxidant activities of the plant were also evaluated by standard methods. Results showed alkaloids, tannins, flavonoids, saponins phenolics in the methanolic extract of Solenostemon monostachyus. Cardiac glycosides and steroids absent in the methanolic extract were found present in the dichloromethane extract. The quantitative phytochemicals recorded were alkaloids (18.05 ± 0.87%), saponins (11.3 ± 0.56 %), tannins (3.3 ± 0.19 GAE). The antioxidant activity testing showed that the methanolic extract has higher DPPH scavenging ability over dichloromethane extract and standard ascorbic acid. In the antimicrobial activity testing, the cup-plate diffusion method was used and the result showed that the dichloromethane extract inhibited the growth of Bacillus species at concentrations 100 mg/ml, 50 mg/ml, 20 mg/ml and 10 mg/ml while methanolic extract inhibited the growth of bacillus species only at 100 mg/ml with 5 mm zone of inhibition. However, both methanol and dichloromethane extract showed no antimicrobial activity on the other test organisms such as Psudomonas aeruginosa, Escherichia coli and Staphylococcus aureus. In conclusion both methanolic and dichloromethane extracts of Solenostemon monostachyus are potential sources of antimicrobial and antioxidant.

Na+/K+-ATPase Revisited: On Its Mechanism of Action, Role in Cancer, and Activity Modulation.

Maintenance of Na+ and K+ gradients across the cell plasma membrane is an essential process for mammalian cell survival. An enzyme responsible for this process, sodium-potassium ATPase (NKA), has been currently extensively studied as a potential anticancer target, especially in lung cancer and glioblastoma. To date, many NKA inhibitors, mainly of natural origin from the family of cardiac steroids (CSs), have been reported and extensively studied. Interestingly, upon CS binding to NKA at nontoxic doses, the role of NKA as a receptor is activated and intracellular signaling is triggered, upon which cancer cell death occurs, which lies in the expression of different NKA isoforms than in healthy cells. Two major CSs, digoxin and digitoxin, originally used for the treatment of cardiac arrhythmias, are also being tested for another indication—cancer. Such drug repositioning has a big advantage in smoother approval processes. Besides this, novel CS derivatives with improved performance are being developed and evaluated in combination therapy. This article deals with the NKA structure, mechanism of action, activity modulation, and its most important inhibitors, some of which could serve not only as a powerful tool to combat cancer, but also help to decipher the so-far poorly understood NKA regulation.

Na+, K+-ATPase α Isoforms and Endogenous Cardiac Steroids in Prefrontal Cortex of Bipolar Patients and Controls

Bipolar disorder is a chronic multifactorial psychiatric illness that affects the mood, cognition, and functioning of about 1–2% of the world’s population. Its biological basis is unknown, and its treatment is unsatisfactory. The α1, α2, and α3 isoforms of the Na+, K+-ATPase, an essential membrane transporter, are vital for neuronal and glial function. The enzyme and its regulators, endogenous cardiac steroids like ouabain and marinobufagenin, are implicated in neuropsychiatric disorders, bipolar disorder in particular. Here, we address the hypothesis that the α isoforms of the Na+, K+-ATPase and its regulators are altered in the prefrontal cortex of bipolar disease patients. The α isoforms were determined by Western blot and ouabain and marinobufagenin by specific and sensitive immunoassays. We found that the α2 and α3 isoforms were significantly higher and marinobufagenin levels were significantly lower in the prefrontal cortex of the bipolar disease patients compared with those in the control. A positive correlation was found between the levels of the three α isoforms in all samples and between the α1 isoform and ouabain levels in the controls. These results are in accordance with the notion that the Na+, K+-ATPase-endogenous cardiac steroids system is involved in bipolar disease and suggest that it may be used as a target for drug development.

English

The antibacterial effect of the leaf, bark and leaf and bark combined of Alstonia boonei on bacterial pathogens, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Streptococcus pneumonia, Proteus mirabilis, Klebsiella pneumonia, Shigella dysenteriae and Salmonella typhimurium was determined using agar diffusion technique to investigate their efficacy as anti-bacterial agent. The phytochemical components of the leaf and bark were studied. The phytochemical screening of the ethanolic leaf extract indicated the presence of tannins, phlobatannins, alkaloids, and cardiac glycosides, reducing sugar, saponins, anthraquinones and steroids, while the bark contains alkaloids, flavonoids, saponins, phlobatannins, anthraquinones, steroids and reducing sugars. The extracts showed varying inhibitory effect against P. aeruginosa and K. pneumonia. S. typhimurium (13±0.27 mm at 100 mg/ml for the leaf extract) being the most susceptible at all concentrations, 12±0.21 and 12±0.62 mm at concentrations 100 and 50 mg/ml respectively for bark extract and similarly for the leaf + bark extract. The result of the study suggests that the extracts of the different component parts of the plant can be used for the treatment of infections caused by the test organisms. Key words: In vitro, antibacterial, Alstonia boonei, extracts, agar diffusion.

Genus Periploca (Apocynaceae): A Review of Its Classification, Phytochemistry, Biological Activities and Toxicology.

The genus Periploca belongs to the family Apocynaceae, which is composed of approximately ten species of plants according to incomplete statistics. Most of these plants serve as folk medicines with a long history, especially Periploca sepium and Periploca forrestii. The botanical classifications, chemical constituents, biological activities and toxicities of the genus Periploca were summarized in the literature from 1897 to early 2019. Though the botanical classification of this genus is controversial, these species are well-known to be rich sources of diverse and complex natural products—above all, cardiac steroids and C21 pregnane steroids with special structures and obvious pharmacological activities. The various crude extracts and 314 isolated metabolites from this genus have attracted much attention in intensive biological studies, indicating that they are equipped with cardiotonic, anti-inflammatory, immunosuppressive, antitumor, antimicrobial, antioxidant, insecticidal and other properties. It is noteworthy that some cardiac glycosides showed hepatotoxicity and cardiotoxicity at certain doses. Therefore, in view of the medical and agricultural value of the genus Periploca, in-depth investigations of the pharmacology in vivo, the mechanisms of biological actions, and the pharmacokinetics of the active ingredients should be carried out in the future. Moreover, in order to ensure the safety of clinical medication, the potential toxicities of cardiac glycosides or other compounds should also be paid attention. This systematic review provides an important reference base for applied research on pharmaceuticals and pesticides from this genus.

Na+, K+-ATPase α3 isoform in frontal cortex GABAergic neurons in psychiatric diseases

Na+, K+-ATPase is an essential membrane transporter. In the brain, the α3 isoform of Na+, K+-ATPase is vital for neuronal function. The enzyme and its regulators, endogenous cardiac steroids (ECS), were implicated in neuropsychiatric disorders. GABAergic neurotransmission was also studied extensively in diseases such as schizophrenia and bipolar disorder (BD). Post mortem brain samples from subjects with depression, schizophrenia or BD and non-psychiatric controls were provided by the Stanley Medical Research Institute. ECS levels were determined by ELISA. Expression levels of the three Na+, K+-ATPase-α isoforms, α1, α2 and α3, were determined by Western blot analysis. The α3 levels in GABAergic neurons in different regions of the brain were quantified by fluorescence immunohistochemistry. The results show that Na+, K+ -ATPase α3 isoform levels were lower in GABAergic neurons in the frontal cortex in BD and schizophrenia as compared with the controls (n = 15 subjects per group). A study on a ‘mini-cohort’ (n = 3 subjects per group) showed that the α3 isoform levels were also lower in GABAergic neurons in the hippocampus, but not amygdala, of bipolar and schizophrenic subjects. In the temporal cortex, higher Na+, K+ -ATPase α3 protein levels were found in the three psychiatric groups. No significant differences in ECS levels were found in this brain area. This is the first report on the distribution of α3 in specific neurons in the human brain in association with mental illness. These results strengthen the hypothesis for the involvement of Na+, K+ -ATPase in neuropsychiatric diseases.

Wallflower (Erysimum cheiri (L.) Crantz) from Past to Future

Wallflower (Erysimum cheiri (L.) Crantz) is a common medicinal plant in Persian medicine and nowadays some traditional products from wallflower are consumed on global markets. The aim of the present study was to study the phytochemical constituents of wallflower and discuss safety evaluations related to the traditional wallflower preparations. Major Persian scholars (e.g. Avicenna) books, Persian manuscripts (e.g. Makhzan-al-advia) and Arabic medical manuscripts (e.g Alshamel-fi alsanaat altebya) of the medieval Islamic era as well as current search engines including Pubmed, Scopus, Siencedirect, and Google Scholar were included in the study from 1700 up to 2018 A.D. In traditional medicine manuscripts, various topical and oral dosage forms of wallflower were administered in low doses. After renaissance, phytochemical investigations reported cardiac steroids in wallflower and it might be the reason that next medical investigations on the herb have been interrupted. According to in vivo studies, topical indications of cardiac steroids in doses lower than their inhibitory concentration 50 (IC50) should be safe and effective in some cutaneous disorders. Wallflower is reported to have several different classes of compounds including: 11 types of cardenolides (such as strophanthidin, bipindogenin, uzarigenin, cannogenol and digitoxygenin derivatives), two flavonoids, a cyanidin and two glucosinolates. Therefore, for safety guarantee, wallflower products require dose adjustment based on IC50 and probable cardenolide soluble content in that dosage forms.