RESUMEN

Triple-negative breast cancer (TNBC) represents around 15% of the 2.26 million breast cancers diagnosed worldwide annually and has the worst outcome. Despite recent therapeutic advances, there remains a lack of targeted therapies for this breast cancer subtype. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor with biological roles in regulating development, xenobiotic metabolism, cell cycle progression and cell death. AhR activation by select ligands can promote tumor suppression in multiple cancer types. AhR can negatively regulate the activity of different oncogenic signaling pathways and can directly upregulate tumor suppressor genes such as p27Kip1. To determine the role of AhR in TNBC, we generated AhR-deficient cancer cells and investigated the impact of AhR loss on TNBC cell growth phenotypes. We found that AhR-deficient MDA-MB-468 TNBC cells have increased proliferation and formed significantly more colonies compared to AhR expressing cells. These cells without AhR expression grew aggressively in vivo. To determine the molecular targets driving this phenotype, we performed transcriptomic profiling in AhR expressing and AhR knockout MDA-MB-468 cells and identified tyrosine receptor kinases, as well as other genes involved in proliferation, survival and clonogenicity that are repressed by AhR. In order to determine therapeutic targeting of AhR in TNBC, we investigated the anti-cancer effects of the novel AhR ligand 11-chloro-7H-benzimidazo[2,1-a]benzo[de]iso-quinolin-7-one (11-Cl-BBQ), which belongs to a class of high affinity, rapidly metabolized AhR ligands called benzimidazoisoquinolines (BBQs). 11-Cl-BBQ induced AhR-dependent cancer cell-selective growth inhibition and strongly inhibited colony formation in TNBC cells.

Asunto(s)

RESUMEN

Triple-negative breast cancer (TNBC) remains a disease with a paucity of targeted treatment opportunities. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that is involved in a wide range of physiological processes, including the sensing of xenobiotics, immune function, development, and differentiation. Different small-molecule AhR ligands drive strikingly varied cellular and organismal responses. In certain cancers, AhR activation by select small molecules induces cell cycle arrest or apoptosis via activation of tumor-suppressive transcriptional programs. AhR is expressed in triple-negative breast cancers, presenting a tractable therapeutic opportunity. Here, we identify a novel ligand of the aryl hydrocarbon receptor that potently and selectively induces cell death in triple-negative breast cancer cells and TNBC stem cells via the AhR. Importantly, we found that this compound, Analog 523, exhibits minimal cytotoxicity against multiple normal human primary cells. Analog 523 represents a high-affinity AhR ligand with potential for future clinical translation as an anticancer agent.

RESUMEN

Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor and functions as a tumour suppressor in different cancer models. In the present study, we report detailed characterization of 11-chloro-7H-benzimidazo[2,1-a]benzo[de]iso-quinolin-7-one (11-Cl-BBQ) as a select modulator of AhR-regulated transcription (SMAhRT) with anti-cancer actions. Treatment of lung cancer cells with 11-Cl-BBQ induced potent and sustained AhR-dependent anti-proliferative effects by promoting G1 phase cell cycle arrest. Investigation of 11-Cl-BBQ-induced transcription in H460 cells with or without the AhR expression by RNA-sequencing revealed activation of p53 signalling. In addition, 11-Cl-BBQ suppressed multiple pathways involved in DNA replication and increased expression of cyclin-dependent kinase inhibitors, including p27Kip1 , in an AhR-dependent manner. CRISPR/Cas9 knockout of individual genes revealed the requirement for both p53 and p27Kip1 for the AhR-mediated anti-proliferative effects. Our results identify 11-Cl-BBQ as a potential lung cancer therapeutic, highlight the feasibility of targeting AhR and provide important mechanistic insights into AhR-mediated-anticancer actions.

Asunto(s)

RESUMEN

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that regulates cell fate via activation of a diverse set of genes. There are conflicting reports describing the role of AhR in cancer. AhR-knockout mice do not develop tumors spontaneously, yet the AhR can act as a tumor suppressor in certain contexts. Loss of tumor suppression by p53 is common in human cancer. To investigate AhR function in the absence of p53, we generated mice lacking both AhR and p53. Mice deficient for AhR and p53 had shortened lifespan, increased tumorigenesis, and an altered tumor spectrum relative to control mice lacking only p53. In addition, knockout of both AhR and p53 resulted in reduced embryonic survival and neonatal fitness. We also examined the consequences of loss of AhR in p53-heterozygous mice and observed a significantly reduced lifespan and enhanced tumor burden. These findings reveal an important role for the AhR as a tumor suppressor in the absence of p53 signaling and support the development of anti-cancer therapeutics that would promote the tumor suppressive actions of the AhR.

Asunto(s)

RESUMEN

p27Kip1 functions to coordinate cell cycle progression through the inhibition of cyclin-dependent kinase (CDK) complexes. p27Kip1 also exerts distinct activities beyond CDK-inhibition, including functioning as a transcriptional regulator. The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor with diverse biological roles. The regulatory inputs that control AhR-mediated transcriptional responses are an active area of investigation. AhR was previously established as a direct regulator of p27Kip1 transcription. Here, we report the physical interaction of AhR and p27Kip1 and show that p27Kip1 expression negatively regulates AhR-mediated transcription. p27Kip1 knockout cells display increased AhR nuclear localisation and significantly higher expression of AhR target genes. This work thus identifies new regulatory cross-talk between p27Kip1 and AhR.

Asunto(s)

RESUMEN

3,3'-Diindolylmethane (DIM), a major phytochemical derived from ingestion of cruciferous vegetables, is also a dietary supplement. In preclinical models, DIM is an effective cancer chemopreventive agent and has been studied in a number of clinical trials. Previous pharmacokinetic studies in preclinical and clinical models have not reported DIM metabolites in plasma or urine after oral dosing, and the pharmacological actions of DIM on target tissues is assumed to be solely via the parent compound. Seven subjects (6 males and 1 female) ranging from 26-65 years of age, on a cruciferous vegetable-restricted diet prior to and during the study, took 2 BioResponse DIM 150-mg capsules (45.3 mg DIM/capsule) every evening for one week with a final dose the morning of the first blood draw. A complete time course was performed with plasma and urine collected over 48 hours and analyzed by UPLC-MS/MS. In addition to parent DIM, two monohydroxylated metabolites and 1 dihydroxylated metabolite, along with their sulfate and glucuronide conjugates, were present in both plasma and urine. Results reported here are indicative of significant phase 1 and phase 2 metabolism and differ from previous pharmacokinetic studies in rodents and humans, which reported only parent DIM present after oral administration. 3-((1H-indole-3-yl)methyl)indolin-2-one, identified as one of the monohydroxylated products, exhibited greater potency and efficacy as an aryl hydrocarbon receptor agonist when tested in a xenobiotic response element-luciferase reporter assay using Hepa1 cells. In addition to competitive phytochemical-drug adverse reactions, additional metabolites may exhibit pharmacological activity highlighting the importance of further characterization of DIM metabolism in humans. SIGNIFICANCE STATEMENT: 3,3'-Diindolylmethane (DIM), derived from indole-3-carbinol in cruciferous vegetables, is an effective cancer chemopreventive agent in preclinical models and a popular dietary supplement currently in clinical trials. Pharmacokinetic studies to date have found little or no metabolites of DIM in plasma or urine. In marked contrast, we demonstrate rapid appearance of mono- and dihydroxylated metabolites in human plasma and urine as well as their sulfate and glucuronide conjugates. The 3-((1H-indole-3-yl)methyl)indolin-2-one metabolite exhibited significant aryl hydrocarbon receptor agonist activity, emphasizing the need for further characterization of the pharmacological properties of DIM metabolites.

Asunto(s)

RESUMEN

Silver nanoparticles (Ag NPs) are among the most common forms of nanoparticles in consumer products, yet the environmental implications of their widespread use remain unclear due to uncertainties about their fate. Because sulfidation of Ag NPs results in the formation of a stable silver sulfide (Ag2S) product, it is likely an important removal mechanism of bioavailable silver in natural waters. In addition to sulfide, the complete conversion of Ag NPs to Ag2S will require dissolved oxygen or some other oxidant so dispersed metal sulfides may be an important pool of reactive sulfide for such reactions in oxygenated systems. The reaction of Ag NPs with zinc sulfide (ZnS) was investigated using a voltammetric method, anodic stripping voltammetry (ASV). ASV provided sensitive, in situ measurements of the release of zinc (Zn2+) cations resulting from the cation exchange reaction between Ag NPs and ZnS. The effects of Ag NP size and surface coatings on the initial rates of sulfidation by ZnS were examined. Sulfidation of smaller Ag NPs generally occurred faster and to a greater extent due to their larger relative surface areas. Sulfidation of Ag NPs capped by citrate and lipoic acid occurred more rapidly relative to polyvinylpyrrolidone (PVP) and branched polyethylene (BPEI). This study demonstrates the utility of voltammetry for such investigations and provides insights into important factors controlling Ag NP sulfidation such as availability of dissolved oxygen, Ag NP size and Ag NP surface coating. Furthermore, this work demonstrates the importance of cation exchange reactions between silver and metal sulfides, and how the environmental release of Ag NPs could alter the speciation of other metals of environmental significance.

RESUMEN

OBJECTIVES: We aimed to compare risk factors for adverse pregnancy outcomes in women living with HIV (WLWH) with those in women of the general population (WGP) in Denmark. Further, we estimated risk of pregnancy- or birth-related complications. METHODS: A retrospective cohort study including all WLWH who delivered a live-born child from 2002 to 2014 and WGP, matched by origin, age, year and parity, was carried out. We compared risk factors during pregnancy and estimated risk of pregnancy- and birth-related complications using multivariate logistic regression. RESULTS: A total of 2334 pregnancies in 304 WLWH and 1945 WGP were included in the study. WLWH had more risk factors present than WGP during pregnancy: previous caesarean section (CS) (24.7% versus 16.3%, respectively; P = 0.0001), smoking (14.2% versus 7.5%, respectively; P = 0.0001) and previous perinatal/neonatal death (2.3% versus 0.9%, respectively; P = 0.03). We found no difference between groups regarding gestational diabetes, hypertensive disorders, low birth weights or premature delivery. More children of WLWH had intrauterine growth retardation (IUGR) [adjusted odds ratio (aOR) 1.9; 95% confidence interval (CI) 1.1-3.2; P = 0.02]. Median gestational age and birth weight were lower in children born to WLWH. WLWH had a higher risk of emergency CS (EmCS) (aOR 1.6; 95% CI 1.2-2.1; P = 0.0005) and postpartum haemorrhage (aOR 1.4; 95% CI 1.0-1.9; P = 0.02) but not infection, amniotomy, failure to progress, low activity-pulse-grimace-appearance-respiration (APGAR) score or signs of asphyxia. CONCLUSIONS: WLWH had more risk factors present during pregnancy, similar risks of most pregnancy- and birth-related complications but a higher risk of postpartum haemorrhage and EmCS compared with WGP. Children born to WLWH had lower median birth weights and gestational ages and were at higher risk of IUGR.

Asunto(s)

RESUMEN

Both clinicians and neuroscientists have been long interested in the topic of fear conditioning, with recent advances in neuroscience, in particular, igniting a shared interest in further translation between these domains. Here, we review some historical aspects of this relationship and the progress that has been made in translating the neuroscientific study of fear conditioning to the conceptualization and treatment of mental disorders, especially anxiety-related disorders. We also address some conceptual and methodological challenges faced by this research, and offer some suggestions to support future progress in the field.

Asunto(s)

RESUMEN

Learning to predict threat is a fundamental ability of many biological organisms, and a laboratory model for anxiety disorders. Interfering with such memories in humans would be of high clinical relevance. On the basis of studies in cell cultures and slice preparations, it is hypothesised that synaptic remodelling required for threat learning involves the extracellular enzyme matrix metalloproteinase (MMP) 9. However, in vivo evidence for this proposal is lacking. Here we investigate human Pavlovian fear conditioning under the blood-brain barrier crossing MMP inhibitor doxycyline in a pre-registered, randomised, double-blind, placebo-controlled trial. We find that recall of threat memory, measured with fear-potentiated startle 7 days after acquisition, is attenuated by ~60% in individuals who were under doxycycline during acquisition. This threat memory impairment is also reflected in increased behavioural surprise signals to the conditioned stimulus during subsequent re-learning, and already late during initial acquisition. Our findings support an emerging view that extracellular signalling pathways are crucially required for threat memory formation. Furthermore, they suggest novel pharmacological methods for primary prevention and treatment of posttraumatic stress disorder.

Asunto(s)

RESUMEN

The active transport of fluids by pumps plays an essential role in engineering and biology. Due to increasing energy costs and environmental issues, topics like noise reduction, increase of efficiency and enhanced robustness are of high importance in the development of pumps in engineering. The study compares pumps in biology and engineering and assesses biomimetic potentials for improving man-made pumping systems. To this aim, examples of common challenges, applications and current biomimetic research for state-of-the art pumps are presented. The biomimetic research is helped by the similar configuration of many positive displacement pumping systems in biology and engineering. In contrast, the configuration and underlying pumping principles for fluid dynamic pumps (FDPs) differ to a greater extent in biology and engineering. However, progress has been made for positive displacement as well as for FDPs by developing biomimetic devices with artificial muscles and cilia that improve energetic efficiency and fail-safe operation or reduce noise. The circulatory system of vertebrates holds a high biomimetic potential for the damping of pressure pulsations, a common challenge in engineering. Damping of blood pressure pulsation results from a nonlinear viscoelastic behavior of the artery walls which represent a complex composite material. The transfer of the underlying functional principle could lead to an improvement of existing technical solutions and be used to develop novel biomimetic damping solutions. To enhance efficiency or thrust of man-made fluid transportation systems, research on jet propulsion in biology has shown that a pulsed jet can be tuned to either maximize thrust or efficiency. The underlying principle has already been transferred into biomimetic applications in open channel water systems. Overall there is a high potential to learn from nature in order to improve pumping systems for challenges like the reduction of pressure pulsations, increase of jet propulsion efficiency or the reduction of wear.

Asunto(s)

RESUMEN

The Publisher regrets that this article is an accidental duplication of an article that has already been published, http://dx.doi.org/10.1016/j.chemphyslip.2015.01.001. The duplicate article has therefore been withdrawn. The full Elsevier Policy on Article Withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.

RESUMEN

We have previously shown that the oxidized cholesterol 3ß-hydroxy-5-oxo-5,6-secocholestan-6-al (atheronal A) reacts covalently with the free amino group of phosphatidylethanolamine (PE) or phosphatidylserine (PS) to produce a Schiff base. Accompanying this interaction, the biophysical properties of the phospholipid membranes are also changed. In the present report, we extend our earlier study of the rate of Schiff base formation in dimyristoyl PE/atheronal A binary mixtures to the more biologically relevant case in which varying amounts of cholesterol are also present. Using optical spectroscopy to monitor reaction kinetics, we demonstrate that the presence of cholesterol reduces the accessibility of the aldehyde moiety of the atheronal A to the free headgroup amine. We also find that the presence of atheronal A promotes the early onset of cholesterol crystallization in the ternary mixtures, perhaps with the Schiff base serving as a site for heterogeneous nucleation.

Asunto(s)

RESUMEN

In vivo, cells reside in a complex environment regulating their fate and function. Most of this complexity is lacking in standard in vitro models, leading to readouts falling short of predicting the actual in vivo situation. The use of engineering tools, combined with deep biological knowledge, leads to the development and use of bioreactors providing biologically sound niches. Such bioreactors offer new tools for biological research, and are now also entering the field of cancer research. Here we present the development and validation of a modular bioreactor system providing: (i) high throughput analyses, (ii) a range of biological conditions, (iii) high degree of control, and (iv) application of physiological stimuli to the cultured samples. The bioreactor was used to engineer a three-dimensional (3D) tissue model of cancer, where the effects of mechanical stimulation on the tumor phenotype were evaluated. Mechanical stimuli applied to the engineered tumor model activated the mechanotransduction machinery and resulted in measurable changes of mRNA levels towards a more aggressive tumor phenotype.

Asunto(s)

RESUMEN

BACKGROUND AND PURPOSE: In acute stroke, CTP is often used to visualize the endangered brain areas, including the ischemic core and the penumbra. Our goal was to assess the evolution of the infarct after mechanical thrombectomy and to analyze the interventional factors determining the fate of the penumbra. MATERIALS AND METHODS: All patients receiving mechanical thrombectomy in the anterior circulation and receiving CTP beforehand were identified. The infarct volume was specified. The clinical parameters, outcome, and interventional results were correlated with the CTP and the final infarct size. RESULTS: In total, 73 patients were included. After mechanical thrombectomy, 78.1% reached a TICI score of 3/2b. The final infarct volume was significantly smaller, with a TICI score of 3/2b compared with less sufficient recanalization (19.60 ± 3 cm(3) versus 38.1 ± 9 cm(3); P < .001). After TICI 3/2b recanalization, 81% ± 5.2% of the potential infarct size (calculated as the sum of infarct core and penumbra) could be rescued. In patients with TICI scores of 2a or worse, only 39 ± 28.3 were salvaged (P < .001). The Alberta Stroke Program Early CT Score after successful recanalization TICI score of 3/2b resulted in a decline of 1.9 ± 1.4 compared with the significantly higher degradation score of 3.7 ± 1.7 after recanalization, with a TICI score of 2a or worse. A recanalization TICI score of 3/2b resulted in an NIHSS improvement of 7.3 ± 0.8 NIHSS points, whereas a poorer recanalization improved on the NIHSS by only 2.5 ± 1.5 points (P < .01). CONCLUSIONS: Mechanical thrombectomy is a potent method to rescue large areas of penumbra in acute stroke.

Asunto(s)

RESUMEN

Using differential scanning calorimetry and small and wide angle X-ray diffraction, we show that, following extended incubation at room temperature, methanol, propanol, and three of the isomers of butanol can induce ordering in dipalmitoyl phosphatidylserine (DPPS) gel phase bilayers. The organization of the bilayers in the presence of ethanol, described previously, is now observed to be a general effect of short chain alcohols. Evidence is presented for tilting of the acyl chains with respect to the bilayer normal in the presence of ethanol or propanol. However, the different chain lengths of the alcohols, and isomeric form, influence the thermal stability of the ordered gel to different extents. This behavior is unlike that of the gel state phosphatidylcholine analog which, in the presence of short chain alcohols, undergoes hydrocarbon chain interdigitation. Dipalmitoyl phosphatidylcholine added to DPPS in the presence of 20 vol% ethanol, acts to suppress the ordered gel phase.

Asunto(s)

RESUMEN

BACKGROUND: Specific immunotherapy via the subcutaneous or oral route is associated with local and, in some cases, systemic side effects and suffers from low patient compliance. Due to its unique immunological features, the skin represents a promising target tissue for effective and painless treatment of type I allergy. The current study was performed to compare the efficacy of transcutaneous immunotherapy via laser-generated micropores to subcutaneous injection. METHODS: BALB/c mice were sensitized by intraperitoneal injection of recombinant grass pollen allergen Phl p 5 together with alum. Subsequently, lung inflammation was induced by repeated intranasal challenge. During the treatment phase, adjuvant-free Phl p 5 was applied in solution to microporated skin or was subcutaneously injected. Lung function and cellular infiltration; Phl p 5-specific serum levels of IgG1, IgG2a, and IgE; and cytokine levels in bronchoalveolar lavage fluids as well as in supernatants of splenocyte cultures were assessed. RESULTS: Both therapeutic approaches reduced airway hyperresponsiveness and leukocyte infiltration into the lungs. Whereas subcutaneous immunotherapy induced a systemic increase in Th2-associated cytokine secretion, transcutaneous application revealed a general downregulation of Th1/Th2/Th17 responses. Successful therapy was associated with induction of IgG2a and an increase in FOXP3+ CD4+ T cells. CONCLUSIONS: Transcutaneous immunotherapy via laser microporation is equally efficient compared with conventional subcutaneous treatment but avoids therapy-associated boosting of systemic Th2 immunity. Immunotherapy via laser-microporated skin combines a painless application route with the high efficacy known from subcutaneous injections and therefore represents a promising alternative to established forms of immunotherapy.

Asunto(s)

RESUMEN

Adaptive behavior often exploits generalizations from past experience by applying them judiciously in new situations. This requires a means of quantifying the relative importance of prior experience and current information, so they can be balanced optimally. In this study, we ask whether the brain generalizes in an optimal way. Specifically, we used Bayesian learning theory and fMRI to test whether neuronal responses reflect context-sensitive changes in ambiguity or uncertainty about experience-dependent beliefs. We found that the hippocampus expresses clear ambiguity-dependent responses that are associated with an augmented rate of learning. These findings suggest candidate neuronal systems that may be involved in aberrations of generalization, such as over-confidence.

Asunto(s)

RESUMEN

An oxidized form of cholesterol, atheronal, is a form found in vivo that has been associated with human pathologies. We have studied mixtures of this oxidized sterol with the phospholipids phosphatidylethanolamine and phosphatidylcholine. We used phospholipids either with palmitoyl and oleoyl acyl chains on the C1 and C2 carbon atoms of glycerol or with both acyl chains being palmitoleoyl. We also compared the effects of atheronal on the curvature properties of these lipids with the action of cholesterol. We studied the bilayer to hexagonal phase transition temperature of mixtures of these lipids using differential scanning calorimetry as well as the dimensions of the hexagonal phase cylinders using X-ray diffraction. Disordering of the lamellar phase was also qualitatively assessed by the loss of sharp diffraction peaks. Our results demonstrate that the modulation of membrane curvature in these systems depends not only on the nature of the sterol, but also on the acyl chain composition of the phospholipids used. In addition, some of the effects of atheronal could be ascribed to reaction of the aldehyde and ketone groups of this oxidized sterol with the amino group of phosphatidylethanolamine.

Asunto(s)

RESUMEN

The objective of this study was to examine the impact of mechanical loading on solute transport in porcine temporomandibular joint (TMJ) discs using the electrical conductivity method. The electrical conductivity, as well as ion diffusivity, of TMJ discs was determined under confined compression with 3 strains in 5 disc regions. The average electrical conductivity over the 5 regions (mean ± SD) at 0% strain was 3.10 ± 0.68 mS/cm, decreased to 2.76 ± 0.58 mS/cm (-11.0%) at 10% strain, and 2.38 ± 0.55 mS/cm (-22.2%) at 20% compressive strain. Correspondingly, the average relative ion diffusivity (mean ± SD) at 0% strain was 0.273 ± 0.055, decreased to 0.253 ± 0.048 (-7.3%) at 10% strain, and 0.231 ± 0.048 (-15.4%) at 20% compressive strain. These results indicated that compressive strain impeded solute transport in the TMJ disc. Furthermore, our results showed that the transport properties of TMJ discs were region-dependent. The electrical conductivity and ion diffusivity in the anterior region were significantly higher than in the posterior region. This regional difference is likely due to the significant differences of tissue hydration between these 2 regions. This study provides important insight into the electrical and solute transport behaviors in TMJ discs under mechanical loading and aids in the understanding of TMJ pathophysiology related to tissue nutrition.

Asunto(s)