Cellular Action Research Articles

Istaroxime treatment ameliorates calcium dysregulation in a zebrafish model of phospholamban R14del cardiomyopathy

The heterozygous Phospholamban p.Arg14del mutation is found in patients with dilated or arrhythmogenic cardiomyopathy. This mutation triggers cardiac contractile dysfunction and arrhythmogenesis by affecting intracellular Ca2+ dynamics. Little is known about the physiological processes preceding induced cardiomyopathy, which is characterized by sub-epicardial accumulation of fibrofatty tissue, and a specific drug treatment is currently lacking. Here, we address these issues using a knock-in Phospholamban p.Arg14del zebrafish model. Hearts from adult zebrafish with this mutation display age-related remodeling with sub-epicardial inflammation and fibrosis. Echocardiography reveals contractile variations before overt structural changes occur, which correlates at the cellular level with action potential duration alternans. These functional alterations are preceded by diminished Ca2+ transient amplitudes in embryonic hearts as well as an increase in diastolic Ca2+ level, slower Ca2+ transient decay and longer Ca2+ transients in cells of adult hearts. We find that istaroxime treatment ameliorates the in vivo Ca2+ dysregulation, rescues the cellular action potential duration alternans, while it improves cardiac relaxation. Thus, we present insight into the pathophysiology of Phospholamban p.Arg14del cardiomyopathy.

Comprehensive chemical proteomics analyses reveal that the new TRi-1 and TRi-2 compounds are more specific thioredoxin reductase 1 inhibitors than auranofin

Anticancer drugs that target cellular antioxidant systems have recently attracted much attention. Auranofin (AF) is currently evaluated in several clinical trials as an anticancer agent that targets the cytosolic and mitochondrial forms of the selenoprotein thioredoxin reductase, TXNRD1 and TXNRD2. Recently, two novel TXNRD1 inhibitors (TRi-1 and TRi-2) have been developed that showed anticancer efficacy comparable to AF, but with lower mitochondrial toxicity. However, the cellular action mechanisms of these drugs have not yet been thoroughly studied. Here we used several proteomics approaches to determine the effects of AF, TRi-1 and TRi-2 when used at IC50 concentrations with the mouse B16 melanoma and LLC lung adenocarcinoma cells, as these are often used for preclinical mouse models in evaluation of anticancer drugs. The results demonstrate that TRi-1 and TRi-2 are more specific TXNRD1 inhibitors than AF and reveal additional AF-specific effects on the cellular proteome. Interestingly, AF triggered stronger Nrf2-driven antioxidant responses than the other two compounds. Furthermore, AF affected several additional proteins, including GSK3A, GSK3B, MCMBP and EEFSEC, implicating additional effects on glycogen metabolism, cellular differentiation, inflammatory pathways, DNA replication and selenoprotein synthesis processes. Our proteomics data provide a resource for researchers interested in the multidimensional analysis of proteome changes associated with oxidative stress in general, and the effects of TXNRD1 inhibitors and AF protein targets in particular.

Age-dependent response to T4 overtreatment and recovery on systemic and organ level.

Thyroid hormone (TH) metabolism and cellular TH action are influenced by ageing. To investigate the response to thyroxine (T4) overtreatment, a kinetic study was conducted in young and aged mice with chronic hyperthyroidism and hormone withdrawal. Five and 22 months old male mice were treated with T4 or PBS over 5 weeks, followed by observation for up to 12 days. Serial analysis was performed for thyroid function parameters, transcript levels of TH target genes, deiodinase type 1 (DIO1) activity as well as serum lipids at 12, 24, 72, 144, 216, and 288 h after cessation of T4 administration. Higher FT3 concentrations and higher renal DIO1 activities were noted in aged mice 12 h after T4 withdrawal and marked thyroid-stimulating hormone elevation was found in aged mice after 12 days compared to respective controls. A biphasic expression pattern occurred for TH target genes in all organs and a hypothyroid organ state was observed at the end of the study, despite normalization of TH serum concentrations after 72 h. In line with this, mirror-image kinetics were detected for serum cholesterol and triglycerides in aged and young mice. Recovery from TH overtreatment in mice involves short- and medium-term adaption of TH metabolism on systemic and organ levels. Increased renal DIO1 activity may contribute to higher T3 concentrations and prolonged thyrotoxicosis followed by hypothyroidism in an aged-mouse organism. Translation of these findings in the clinical setting seems warranted and may lead to better management of hyperthyroidism and prevention of T4 overtreatment in aged patients.

Repolarization heterogeneity within the myocardial infarction border zone correlates with variability of myocyte remodeling

Abstract Background Myocardial infarction (MI) results in a regional scar, with a border zone (BZ) of surviving myocytes interspersed with fibrosis providing an anatomical substrate for re-entry. Heterogeneous repolarization within the BZ may add a functional component aggravating re-entrant arrhythmias. Purpose We studied BZ heterogeneity and developed novel methodology for high resolution mapping of local in vivo activation-repolarization intervals (ARI) within the BZ and for studying the relation to cellular action potential (AP) profiles of cells isolated from the BZ. Methods Anterior-septal myocardial infarction was induced in 5 domestic pigs by 120-minute occlusion of the left anterior descending artery followed by reperfusion (18.9±4.7% of the left ventricle). After 1-month, electro-anatomical mapping was performed. Contact mapping was used to define the BZ (bipolar voltage 0.5–1.5mV). A non-contact recording of a 64-electrode array was translated to 2048 non-contact electrograms distributed over the LV. The non-contact electrograms were processed to determine the ARIs using a custom-made algorithm, validated against monophasic action potential recordings. After 2–4 days recovery, single cardiomyocytes were enzymatically isolated from the anterior-septal BZs and remote regions. Cardiomyocytes were field stimulated at 1Hz at 37°C and cellular AP duration (APD) was optically recorded (fluorescent voltage-sensitive dye Di-8-Annepps). Results In vivo, regional ARIs tended to be longer in the BZs than remote. ARI heterogeneity, quantified as the standard deviation of ARIs in a neighborhood of 1cm radius, was increased in the BZ (anterior BZ: 3.4±1.0 ms, P=0.052, septal BZ: 3.6±1.7 ms, P=0.027 vs remote: 2.0±0.5 ms). Cellular APD was measured in large population samples (>100 cells per region in each pig) and was longer in BZ myocytes compared to the remote region. Cellular APD heterogeneity, measured as the standard deviation within cell population samples pooled by region per animal, was increased in the BZ (anterior BZ: 105.9±17.0 ms, P=0.0010; septal BZ: 98.1±20.8 ms, P=0.0127 vs remote: 73.9±8.6 ms). Cell APD correlated to in vivo ARI (R2=0.34, P=0.021) and cellular heterogeneity correlated strongly with in vivo heterogeneity (R2=0.67, P=0.002). Conclusion In the BZ of MI, in vivo regional heterogeneity adds a functional substrate for re-entry that may result from heterogeneous cellular remodeling and increased cell-cell APD variability. Funding Acknowledgement Type of funding sources: Public Institution(s). Main funding source(s): KU Leuven BOF-C1 “Blood pressure induced premature ventricular beats as triggers for ventricular arrhythmia in ischemic cardiomyopathy”

Limitations of drug concentrations used in cell culture studies for understanding clinical responses of NSAIDs.

In this review, the in vitro cellular effects of six nonsteroidal anti-inflammatory drugs (NSAIDs), salicylate, ibuprofen, naproxen, indomethacin, celecoxib and diclofenac, are examined. Inhibition of prostanoid synthesis in vitro generally occurs within the therapeutic range of plasma concentrations that are observed in vivo, consistent with the major action of NSAIDs being inhibition of prostanoid production. An additional probable cellular action of NSAIDs has been discovered recently, viz. decreased oxidation of the endocannabinoids, 2-arachidonoyl glycerol and arachidonyl ethanolamide. Many effects of NSAIDs, other than decreased oxidation of arachidonic acid and endocannabinoids, have been put forward but almost all of these additional processes are observed at supratherapeutic concentrations when the concentration of albumin, the major protein that binds NSAIDs, is taken into account. However, one exception is salicylate, a very potent inhibitor of the neutrophilic enzyme, myeloperoxidase, the inhibition of which leads to reduced production of the inflammatory mediator, hypochlorous acid, and inhibition of the inflammation associated with rheumatoid arthritis.

Evaluation of the use of advanced ozone oxidative process in reducing the danger of environmental toxicity by endocrine interferences of magistral pharmacy

The presence of emerging pollutants in the waters has been observed worldwide, resulting from improper domestic disposal, non-recommended veterinarian use, and product waste from pharmaceutical industries and magistral pharmacies. The contamination provoked, besides causing damage to the environment, remains in potable water even after passing through the treatment plants. The objective of this work was to verify the existence of environmental toxicity of raw effluents from gross pharmacy laboratories, as well as the same effluent treated with POA via ozone in the time of 1 hour, having as a risk parameter the changes that they cause in Euglena gracilis algae. Photosynthetic efficiency tests were conducted via PAM, and chlorophyll concentration and behavioral evaluation were checked via NGTOX. The results demonstrate that the hormone laboratory was considered the most impacted effluent treated, with lower production and significant chlorophyll reduction. It presented reduction in photosynthetic post-ozonation activity, due to the hormone decomposition, oxidative potential and ethylene formation. Effluents from psychotropic and solid laboratories presented different production demand, but similar follow-up, with impact on the behavior and algae’s photosynthetic activity, due to the presence of active substances on cellular action potentials. The treated effluent from dermocosmetics laboratory influenced the chlorophyll concentration, as well as the general speed and velocity of surface ascent. The behavioral differences between the laboratories and the pre and post-ozonation conditions demonstrate that the effluent treatment should be distinguished, according to the characteristics of the manipulated substances in each laboratory.

Biological potential of apigetrin in the medicine for the treatment of various forms of inflammatory disorders: Phytotherapeutic approach for respiratory infections

Apigetrin is apigenin-7-O-glucoside is a natural flavonoid glucoside also called flavonoidal class plant derive phytochemical found to be present in the nature in Matricaria chamomilla, Scutellaria baicalensis, Stachys tibetica and Teucrium gnaphalodes. In order to know the medicinal potential of apigetrin in the medicine and health sectors, here numerous scientific data have been collected from various literature sources and analyzed for their health beneficial potential. Medicinal importance and pharmacological activities data of apigetrin have been collected from various literature sources and analyzed for their therapeutic potential in the medicine. Molecular mechanism of apigetrin in different disorders data have been also collected from various literature sources and analyzed for their cellular action. All the collected scientific data have been properly analyzed to get the better result in the present investigation. Scientific data of various literature sources revealed the biological potential of apigetrin in the medicine for the treatment of numerous health complications. From the data analysis of different scientific work of the literature sources it was found that apigetrin have neuroprotective potential due to their anti-inflammatory activity and inhibitory potential on oxidative stresses. Further biological effect on lipopolysaccharides (LPS)-induced otitis media showed its anti-inflammatory and anti-oxidant potential. Literature data analysis signified the biological role of TNF-α for their anti-inflammatory activity. Literature database analysis also revealed their biological importance to treat inflammatory diseases of upper respiratory infections. Scientific data analysis of various literature works revealed the biological potential of apigetrin in the medicine for the treatment of various form of inflammatory disorders.

Ten-Year Simulation of the Effects of Denosumab on Bone Remodeling in Human Biopsies.

ABSTRACTPostmenopausal osteoporosis is a disease manifesting in degradation of bone mass and microarchitecture, leading to weakening and increased risk of fracture. Clinical trials are an essential tool for evaluating new treatments and may provide further mechanistic understanding of their effects in vivo. However, the histomorphometry from clinical trials is limited to 2D images and reflects single time points. Biochemical markers of bone turnover give global insight into a drug's action, but not the local dynamics of the bone remodeling process and the cells involved. Additionally, comparative trials necessitate separate treatment groups, meaning only aggregated measures can be compared. In this study, in silico modeling based on histomorphometry and pharmacokinetic data was used to assess the effects of treatment versus control on μCT scans of the same biopsy samples over time, matching the changes in bone volume fraction observed in biopsies from denosumab and placebo groups through year 10 of the FREEDOM Extension trial. In the simulation, treatment decreased osteoclast number, which led to a modest increase in trabecular thickness and osteocyte stress shielding. Long‐term bone turnover suppression led to increased RANKL production, followed by a small increase in osteoclast number at the end of the 6‐month–dosing interval, especially at the end of the Extension study. Lack of treatment led to a significant loss of bone mass and structure. The study's results show how in silico models can generate predictions of denosumab cellular action over a 10‐year period, matching static and dynamic morphometric measures assessed in clinical biopsies. The use of in silico models with clinical trial data can be a method to gain further insight into fundamental bone biology and how treatments can perturb this. With rigorous validation, such models could be used for informing the design of clinical trials, such that the number of participants could be reduced to a minimum to show efficacy. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

DEPIS: A combined dielectrophoresis and impedance spectroscopy platform for rapid cell viability and antimicrobial susceptibility analysis

Antimicrobial resistance (AMR) is caused by inappropriate or excessive antibiotic consumption. Early diagnosis of bacterial infections can greatly curb empirical treatment and thus AMR. Current diagnostic procedures are time-consuming as they rely on gene amplification and cell culture techniques that are inherently limited by the doubling rate of the involved species. Further, biochemical methods for species identification and antibiotic susceptibility testing for drug/dose effectiveness take several days and are non-scalable. We report a real-time, label-free approach called DEPIS that combines dielectrophoresis (DEP) for bacterial enrichment and impedance spectroscopy (IS) for cell viability analysis under 60 min. Target bacteria are captured on interdigitated electrodes using DEP (30 min) and their antibiotic-induced stress response is measured using IS (another 30 min). This principle is used to generate minimum bactericidal concentration (MBC) plots by measuring impedance change due to ionic release by dying bacteria in a low conductivity buffer. The results are rapid since they rely on cell death rather than cell growth which is an intrinsically slower process. The results are also highly specific and work across all bactericidal antibiotics studied, irrespective of their cellular target or drug action mechanism. More importantly, preliminary results with clinical isolates show that methicillin-susceptible Staphylococcus aureus (MSSA) can easily be differentiated from methicillin-resistant S. aureus (MRSA) under 1 h. This rapid cell analyses approach can aid in faster diagnosis of bacterial infections and benefit the clinical decision-making process for antibiotic treatment, addressing the critical issue of AMR.

Oat polyphenol avenanthramide-2c confers protection from oxidative stress by regulating the Nrf2-ARE signaling pathway in PC12 cells

Accumulating evidence has demonstrated that cellular antioxidant systems play essential roles in retarding oxidative stress-related diseases, such as Parkinson's disease. Because nuclear factor erythroid 2-related factor 2 (Nrf2) is a chief regulator of cellular antioxidant systems, small molecules with Nrf2-activating ability may be promising neuroprotective agents. Avenanthramide-2c (Aven-2c), avenanthramide-2f (Aven-2f) and avenanthramide-2p (Aven-2p) are the most abundant avenanthramides in oats, and they have been documented to possess multiple pharmacological benefits. In this work, we synthesized these three compounds and evaluated their cytoprotective effect against oxidative stress-induced PC12 cell injuries. Aven-2c displayed the best protective potency among them. Aven-2c conferred protection on PC12 cells by scavenging free radicals and activating the Nrf2-ARE signaling pathway. Pretreatment of PC12 cells with Aven-2c efficiently enhanced Nrf2 nuclear accumulation and evoked the expression of a set of cytoprotective molecules. The mechanistic study also supports that Nrf2 activation is the molecular basis for the cellular action of Aven-2c. Collectively, this study demonstrates that Aven-2c is a potent Nrf2 agonist, shedding light on the potential usage of Aven-2c in the treatment of neuroprotective diseases.

Health Beneficial Potential of Pectolinarigenin on Human Diseases: An Updated Review of Medicinal Importance and Pharmacological Activity

Medicinal Plants are valuable source of phytochemicals which have been used in the medicine as source of raw material since very ancient time in the world. Flavonoids are one of the important classes of phytochemical basically present in the fruits, vegetables, grains, wine, tea etc. Flavonoids also play an important role in the defense mechanism of plants and produce different colours in the plants. Pectolinarigenin is a natural flavonoidal compound having molecular formula C<sub>17</sub>H<sub>14</sub>O<sub>6</sub> and molecular weight 314.28. Pectolinarigenin is present in various plants and examples are Clerodendrum phlomidis, Eupatorium odoratum, Cirsium chanroenicum, Cirsium japonlcum, Chromolaena odorata, Cirsium setidens, and Trollius chinensis. Pectolinarigenin acts as an anticancer agent against various types of human malignancies, such as lung cancer, melanoma, hepatocellular carcinoma and colorectal adenocarcinoma. It has also anti-inflammatory, anti-allergy, cytotoxic and hepatoprotective properties. Pectolinarigenin gained attention from researchers and clinicians due to their anticancer properties and in future it could be the best choice for cancer treatment. The purpose of this review paper is to summarize all the pharmacological properties of pectolinarigenin on cancer and other disorders, describe the mode of action and possible pathways for cellular level action. The present review initially highlights the current status of flavonoids and their pharmaceutical importance, role of pectolinarigenin in human disorders, and in later section, summarizes analytical techniques of pectolinarigenin as lead molecules. This review will support all the ongoing research of pectolinarigenin through out world for their beneficial properties in all the scientific discipline.

Investigational Treatments for COVID-19 may Increase Ventricular Arrhythmia Risk Through Drug Interactions.

Many drugs that have been proposed for treatment of coronavirus disease 2019 (COVID‐19) are reported to cause cardiac adverse events, including ventricular arrhythmias. In order to properly weigh risks against potential benefits, particularly when decisions must be made quickly, mathematical modeling of both drug disposition and drug action can be useful for predicting patient response and making informed decisions. Here, we explored the potential effects on cardiac electrophysiology of four drugs proposed to treat COVID‐19: lopinavir, ritonavir, chloroquine, and azithromycin, as well as combination therapy involving these drugs. Our study combined simulations of pharmacokinetics (PKs) with quantitative systems pharmacology (QSP) modeling of ventricular myocytes to predict potential cardiac adverse events caused by these treatments. Simulation results predicted that drug combinations can lead to greater cellular action potential prolongation, analogous to QT prolongation, compared with drugs given in isolation. The combination effect can result from both PK and pharmacodynamic drug interactions. Importantly, simulations of different patient groups predicted that women with pre‐existing heart disease are especially susceptible to drug‐induced arrhythmias, compared with diseased men or healthy individuals of either sex. Statistical analysis of population simulations revealed the molecular factors that make certain women with heart failure especially susceptible to arrhythmias. Overall, the results illustrate how PK and QSP modeling may be combined to more precisely predict cardiac risks of COVID‐19 therapies.

Controllability of voltage- and calcium-driven cardiac alternans in a map model.

Certain cardiac arrhythmias are preceded by electrical alternans, a state characterized by beat-to-beat alternation in cellular action potential duration. Cardiac alternans may arise from different mechanisms including instabilities in voltage or intracellular calcium cycling. Although a number of techniques have been proposed to suppress alternans, these methods have mainly been tested using models that do not support calcium-driven alternans. Therefore, it is important to understand how control methods may perform when alternans is driven by instabilities in calcium cycling. In this study, we applied controllability analysis to a discrete map of alternans dynamics in a cardiac cell. We compared two different controllability measures to determine to what extent different control strategies could suppress alternans and tested these predictions using three feedback controllers. We found a modal controllability measure, unlike the minimum singular value of the controllability matrix, consistently indicated the control strategies requiring the least control effort and yielding the smallest closed-loop eigenvalue. In addition, action potential duration was identified as the most effective variable through which control can be applied, regardless of alternans mechanism, although sarcoplasmic reticulum calcium load was also useful for the calcium-driven alternans cases.

Insulin signaling in health and disease.

The molecular mechanisms of cellular insulin action have been the focus of much investigation since the discovery of the hormone 100 years ago. Insulin action is impaired in metabolic syndrome, a condition known as insulin resistance. The actions of the hormone are initiated by binding to its receptor on the surface of target cells. The receptor is an α2β2 heterodimer that binds to insulin with high affinity, resulting in the activation of its tyrosine kinase activity. Once activated, the receptor can phosphorylate a number of intracellular substrates that initiate discrete signaling pathways. The tyrosine phosphorylation of some substrates activates phosphatidylinositol-3-kinase (PI3K), which produces polyphosphoinositides that interact with protein kinases, leading to activation of the kinase Akt. Phosphorylation of Shc leads to activation of the Ras/MAP kinase pathway. Phosphorylation of SH2B2 and of Cbl initiates activation of G proteins such as TC10. Activation of Akt and other protein kinases produces phosphorylation of a variety of substrates, including transcription factors, GTPase-activating proteins, and other kinases that control key metabolic events. Among the cellular processes controlled by insulin are vesicle trafficking, activities of metabolic enzymes, transcriptional factors, and degradation of insulin itself. Together these complex processes are coordinated to ensure glucose homeostasis.

Wound Healing by Keratinocytes: A Cytoskeletal Perspective

Skin, being the protective barrier against the environment, can be subject to frequent trauma and stress, hence has the ability to heal itself rapidly. This capacity is attributed to a large number of resident stem cells and progenitors in the skin, which are activated to proliferate, migrate and differentiate to recreate the cellular diversity and regain tissue integrity. The barrier function of the skin is maintained by the epidermis, a multilayered epithelial compartment formed of keratinocytes. Wound repair evokes the capacity of cells to sense and respond to environmental cues. Tissue damage demands rapid cellular action, wherein spatio-temporally regulated cellular responses are critical determinants of the outcome of healing. Hence cells surrounding the wound have to immediately sense the damage and must activate the key signaling pathways to launch the wound-healing response. Emerging data is indicating that mechanical tension release is one of the first cues sensed by the neighboring cells of the damage. This cue is relayed by the cytoskeleton and converted into biochemical and cellular signals, which help the cells to respond accordingly to the trauma. In this review, we will focus on the role of keratinocytes and keratinocyte stem cells in wound healing, and the cytoskeletal dynamics involved therein.

Repeated oral administration of ethanolic extract of Woodfordia fruticosa (L.) Kurz. flowers against the animal models of depression

Background: The incidence of depressive disease has increased in every year; it differs from county to county and risk issue is also differed among the United States and lifestyle change, cultures. Depression is a critical fitness trouble global such as in India. Due to the existing use of antidepressant drugs, there is an expansion of provoking symptoms. Among the most current decade, the disclosure of more comfortable stimulant natural treatment plans is on the rise. Woodfordia fruticosa flowers have been utilized in historic human beings prescription different illnesses incorporating valuable worried device disorders in Ayurveda. Aims and Objectives: The study is undertaken to investigate the antidepressant activity of W. fruticosa flowers using behavioral screening models. Materials and Methods: Freshly prepared ethanolic extract of W. fruticosa (EWF) is treated twice daily for 14 consecutive days. After 60 min of last dose, administration of EWF in animals was subjected to behavioral screening models (forced swim test and tail suspension test) to evaluate antidepressant activity. Results: Our result indicates that oral administration of EWF flowers shown significant (P < 0.0001 [at 1% level]) antidepressant activity by reducing the immobility time in both the models. Conclusion: W. fruticosa flowers exhibit significant antidepressant activity. Further mechanistic studies are required to confirm the cellular action and therapeutic efficacy of EWF against depression.

Study of the effect of natural polyphenols&#x0D; on the prostate cancer Du145 cell cycle

Relevance: Although natural polyphenols are known as&#x0D; useful nutrients, their use is quite occasional. This is probably&#x0D; due to a lack of complete information on their biochemical&#x0D; structure and mechanism of action. Scientific literature offers&#x0D; many studies on polyphenols’ benefits and metabolic effects&#x0D; on the body. Polyphenols are known to improve body function&#x0D; and increase physiological activity. Polyphenols are used in&#x0D; the body’s normal physiological function and metabolic disorders, and their therapeutic effect can be controlled in complex&#x0D; pathological conditions.&#x0D; Therefore, in our study, we used curcumin and carnosic&#x0D; acid to control the natural polyphenols’ metabolic effects’&#x0D; structure and function in prostate cancer. Metabolic disorders&#x0D; in mitochondria, one of the organelles responsible for cell energy metabolism, have an irreversible effect on the cell, organ,&#x0D; and the system as a whole. This urges the study of cellular action on individual organelles, including the biochemical and&#x0D; physiological aspects.&#x0D; The purpose was to determine the effect of natural polyphenols – curcumin and carnosic acid – on the prostate cancer&#x0D; cell cycle.&#x0D; Results: Combination of curcumin and carnosic acid caused&#x0D; an exposure time-dependent suppression of Du145 cancer cells&#x0D; growth at G0/1 and S phases.&#x0D; Conclusions: The effect of natural polyphenols – curcumin&#x0D; and carnosic acid – on the Du145 prostate cancer cell cycle was&#x0D; determined. Curcumin 7μM and carnosic acid 10μM in combination stopped phases G0/1 and S’ prostate cancer cells division

Thyroid Hormone and Deiodination in Innate Immune Cells.

Thyroid hormone has recently been recognized as an important determinant of innate immune cell function. Highly specialized cells of the innate immune system, including neutrophils, monocytes/macrophages, and dendritic cells, are capable of identifying pathogens and initiating an inflammatory response. They can either phagocytose and kill microbes, or recruit other innate or adaptive immune cells to the site of inflammation. Innate immune cells derive from the hematopoietic lineage and are generated in the bone marrow, from where they can be recruited into the blood and tissues in the case of infection. The link between the immune and endocrine systems is increasingly well established, and recent studies have shown that innate immune cells can be seen as important thyroid hormone target cells. Tight regulation of cellular thyroid hormone availability and action is performed by thyroid hormone transporters, receptors, and the deiodinase enzymes. Innate immune cells express all these molecular elements of intracellular thyroid hormone metabolism. Interestingly, there is recent evidence for a causal relationship between cellular thyroid hormone status and innate immune cell function. This review describes the effects of modulation of intracellular thyroid hormone metabolism on innate immune cell function, specifically neutrophils, macrophages, and dendritic cells, with a special focus on the deiodinase enzymes. Although there are insufficient data at this stage for conclusions on the clinical relevance of these findings, thyroid hormone metabolism may partially determine the innate immune response and, by inference, the clinical susceptibility to infections.

Development of a miRNA-controlled dual-sensing system and its application for targeting miR-21 signaling in tumorigenesis

MicroRNAs (miRNAs) are considered to be strong prognostic markers and key therapeutic targets in human diseases, especially cancer. A sensitive monitoring platform for cancer-associated miRNA (oncomiR) action is needed for mechanistic studies, preclinical evaluation, and inhibitor screening. In this study, we developed and systemically applied a sensitive and efficient lentivirus-based system for monitoring oncomiR actions, essentially miR-21. The specificity and sensitivity of “miRDREL” against various oncomiRs were validated by checking for tight correlations between their expression and targeting efficacy. Experiments based on the transfection of synthetic mimics and antagomir-mediated depletion of oncomiRs further confirmed the specificity of the system. Systemic application of miRDRELs to natural oncomiR targets, knockdown of key microprocessors, and physiological triggering of oncomiRs also demonstrated that the system is an effective tool for monitoring cellular oncomiR action. Importantly, molecular modeling-based screening confirmed the action of the miR-21-targeting drug ivermectin and led to the identification of a new effective derivative, GW4064, for inhibiting oncogenic DDX23-miR-21 signaling. Furthermore, proteomic-kinase inhibitor screenings identified a novel oncogenic kinome-DDX23-miR-21 axis and thus expands our understanding of miR-21 targeting therapeutics in tumorigenesis. Taken together, these data indicate that miRDREL and its versatile application have great potential in basic, preclinical studies and drug development pipelines for miRNA-related diseases, especially cancer.

Non-steroidal Anti-inflammatory Drugs Target TWISTED DWARF1-Regulated Actin Dynamics and Auxin Transport-Mediated Plant Development

The widely used non-steroidal anti-inflammatory drugs (NSAIDs) are derivatives of the phytohormone salicylic acid (SA). SA is well known to regulate plant immunity and development, whereas there have been few reports focusing on the effects of NSAIDs in plants. Our studies here reveal that NSAIDs exhibit largely overlapping physiological activities to SA in the model plant Arabidopsis. NSAID treatments lead to shorter and agravitropic primary roots and inhibited lateral root organogenesis. Notably, in addition to the SA-like action, which in roots involves binding to the protein phosphatase 2A (PP2A), NSAIDs also exhibit PP2A-independent effects. Cell biological and biochemical analyses reveal that many NSAIDs bind directly to and inhibit the chaperone activity of TWISTED DWARF1, thereby regulating actin cytoskeleton dynamics and subsequent endosomal trafficking. Our findings uncover an unexpected bioactivity of human pharmaceuticals in plants and provide insights into the molecular mechanism underlying the cellular action of this class of anti-inflammatory compounds.