Membrane-initiated actions of estrogens in neuroendocrinology: emerging principles

Hormonal ligands for the nuclear receptor superfamily have at least two interacting mechanisms of action: 1) classical transcriptional regulation of target genes (genomic mechanisms); and 2) nongenomic actions that are initiated at the cell membrane, which could impact transcription. Although transcriptional mechanisms are increasingly well understood, membrane-initiated actions of these ligands are incompletely understood. Historically, this has led to a considerable divergence of thought in the molecular endocrine field. We have attempted to uncover principles of hormone action that are relevant to membrane-initiated actions of estrogens. There is evidence that the membrane-limited actions of hormones, particularly estrogens, involve the rapid activation of kinases and the release of calcium. Membrane actions of estrogens, which activate these rapid signaling cascades, can also potentiate nuclear transcription. These signaling cascades may occur in parallel or in series but subsequently converge at the level of modification of transcriptionally relevant molecules such as nuclear receptors and/or coactivators. In addition, other hormones or neurotransmitters may also activate cascades to crosstalk with estrogen receptor-mediated transcription. The idea of synergistic coupling between membrane-initiated and genomic actions of hormones fundamentally revises the paradigms of cell signaling in neuroendocrinology.

Acute actions of thyroid hormone on blood vessel biochemistry and physiology

Purpose of review Description of the progress about the vascular effects promoted by thyroid hormones. Recent findings Over the past few years, a number of studies have shown that in addition to genomic effects on blood vessels, thyroid hormones exert extranuclear nongenomic effects on vascular smooth muscle cells and endothelium. These nongenomic effects occur rapidly and do not involve thyroid hormone response elements-mediated transcriptional events. In this context, the genomic and nongenomic events promoted by thyroid hormones act in concert to control the vascular hemodynamic and regulate the cardiovascular function. Summary Considering the antiatherogenic property of thyroid hormones and the rapid effects produced by this molecule as a vasodilator, including that in the coronary bed, a better understanding of the molecular mechanisms involved in its action may contribute to the development of drugs that can be clinically used to increase the known benefits promoted by thyroid hormones in cardiovascular physiology.

Thyroid hormone stimulates NO production via activation of the PI3K/Akt pathway in vascular myocytes

Aims Thyroid hormone (TH) rapidly relaxes vascular smooth muscle cells (VSMCs). However, the mechanisms involved in this effect remain unclear. We hypothesize that TH-induced rapid vascular relaxation is mediated by VSMC-derived nitric oxide (NO) production and is associated with the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signalling pathway. Methods and results NO levels were determined using a NO-specific fluorescent dye (DAF-2) and nitrite (NO(2)) levels. Expression of NO synthase (NOS) isoforms and proteins of the PI3K/Akt pathway was determined by both western blotting and immunocytochemistry. Myosin light chain (MLC) phosphorylation levels were also investigated by western blotting. Exposure of cultured VSMCs from rat thoracic aortas to triiodothyronine (T3) resulted in a significant decrease of MLC phosphorylation levels. T3 also induced a rapid increase in Akt phosphorylation and increased NO production in a dose-dependent manner (0.001-1 mu M). VSMCs stimulated with T3 for 30 min showed an increase in the expression of all three NOS isoforms and augmented NO production, effects that were prevented by inhibitors of PI3K. Vascular reactivity studies showed that vessels treated with T3 displayed a decreased response to phenylephrine, which was reversed by NOS inhibition. These data suggest that T3 treatment induces greater generation of NO both in aorta and VSMCs and that this phenomenon is endothelium independent. In addition, these findings show for the first time that the PI3K/Akt signalling pathway is involved in T3-induced NO production by VSMCs, which occurs with expressive participation of inducible and neuronal NOS. Conclusion Our data strongly indicate that T3 causes NO-dependent rapid relaxation of VSMC and that this effect is mediated by the PI3K/Akt signalling pathway.

T3 acutely increases GH mRNA translation rate and GH secretion in hypothyroid rats

Cytoskeleton controls the stability of transcripts, by mechanisms that involve mRNAs and eEF1A attachment to it. Besides, it plays a key role in protein synthesis and secretion, which seems to be impaired in somatotrophs of hypothyroid rats, whose cytoskeleton is disarranged. This study investigated the: eEF1A and GH mRNA binding to cytoskeleton plus GH mRNA translation rate and GH secretion, in sham-operated and thyroidectomized rats treated with T3 or saline, and killed 30 min thereafter. Thyroidectomy reduced: (a) pituitary F-actin content, and eEF1A plus GH mRNA binding to it; (b) GH mRNA recruitment to polysome; and (c) liver IGF-1 mRNA expression, indicating that GH mRNA stability and translation rate, as well as GH secretion were impaired. T3 acutely reversed all these changes, which points toward a nongenomic action of T3 on cytoskeleton rearrangement, which might contribute to the increase on GH mRNA translation rate and GH secretion. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

New insight into the mechanisms associated with the rapid effect of T-3 on AT1R expression

The angiotensin II type 1 receptor (AT1R) is involved in the development of cardiac hypertrophy promoted by thyroid hormone. Recently, we demonstrated that triiodothyronine (T-3) rapidly increases AT1R mRNA and protein levels in cardiomyocyte cultures. However, the molecular mechanisms responsible for these rapid events are not yet known. In this study, we investigated the T-3 effect on AT1R mRNA polyadenylation in cultured cardiomyocytes as well as on the expression of microRNA-350 (miR-350), which targets AT1R mRNA. The transcriptional and translational actions mediated by T-3 on AT1R levels were also assessed. The total content of ubiquitinated proteins in cardiomyocytes treated with T-3 was investigated. Our data confirmed that T-3 rapidly raised AT1R mRNA and protein levels, as assessed by real-time PCR and western blotting respectively. The use of inhibitors of mRNA and protein synthesis prevented the rapid increase in AT1R protein levels mediated by T-3. In addition, T-3 rapidly increased the poly-A tail length of the AT1R mRNA, as determined by rapid amplification of cDNA ends poly-A test, and decreased the content of ubiquitinated proteins in cardiomyocytes. On the other hand, T-3 treatment increased miR-350 expression. In parallel with its transcriptional and translational effects on the AT1R, T-3 exerted a rapid posttranscriptional action on AT1R mRNA polyadenylation, which might be contributing to increase transcript stability, as well as on translational efficiency, resulting to the rapid increase in AT1R mRNA expression and protein levels. Finally, these results show, for the first time, that T-3 rapidly triggers distinct mechanisms, which might contribute to the regulation of AT1R levels in cardiomyocytes. Journal of Molecular Endocrinology (2012) 49, 11-20

Nongenomic signaling of the retinoid X receptor through binding and inhibiting Gq in human platelets

Retinoid X receptors (RXRs) are important transcriptional nuclear hormone receptors, acting as either homodimers or the binding partner for at least one fourth of all the known human nuclear receptors. Functional nongenomic effects of nuclear receptors are poorly understood; however, recently peroxisome proliferator-activated receptor (PPAR) gamma, PPAR beta, and the glucocorticoid receptor have all been found active in human platelets. Human platelets express RXR alpha, and RXR beta. RXR ligands inhibit platelet aggregation and TXA(2) release to ADP and the TXA(2) receptors, but only weakly to collagen. ADP and TXA(2) both signal via the G protein, Gq. RXR rapidly binds Gq but not Gi/z/o/t/gust in a ligand-dependent manner and inhibits Gq-induced Rac activation and intracellular calcium release. We propose that RXR ligands may have beneficial clinical actions through inhibition of platelet activation. Furthermore, our results demonstrate a novel nongenomic mode for nuclear receptor action and a functional cross-talk between G-protein and nuclear receptor signaling families. (C) 2007 by The American Society of Hematology.

Why, how, what for? : motivations, actions and expectations in habitual entrepreneurship

Principal topic: Is habitual entrepreneurship different? Answering this is important to the field, however there is little systematic evidence, thus far. We addresses this by examining the role experience plays at three possible points of difference: motivations, actions and expectations; and by comparing those currently in the process of starting a business with those who have recent success in business creation. Firstly, we assess the balance of opportunity versus necessity motivation, internally versus externally stimulated decision processes and future growth aspirations. Literature suggests novices are more likely motivated to nascency out of necessity, and favour a manageable business size, while habitual entrepreneurs are more likely motivated by internally stimulated or idea driven processes. Secondly, we examine actions undertaken by successful experienced founders during gestation, contrasting ‘information collection’ and ‘opportunity definition’. Drawing on prior research we expect novices more likely to have enacted ‘information search’ while habitual entrepreneurs enact ‘opportunity definition’. Thirdly, we examine perceptions of venture success, where findings on overconfidence suggest that habitual entrepreneurs expect a higher chance of success for their ventures, while inexperience leads novices to underestimate the difficulty of entrepreneurial survival. Method: Empirical evidence to test these conjectures was drawn from a screened random sample of over 1100 Australian nascent and newly started business ventures. This information was collected during 2007/8 using a telephone survey. Results and Implications: Why do habitual entrepreneurs keep coming back? Findings suggest that while the pursuit of opportunity is shared by novice and experienced entrepreneur alike, consideration of repeat entrepreneurship may be motivated by a desire for growth. While idea driven motivations might not delineate a distinction during nascency, it does seem to be a factor contributing to the success of young firms. This warrants further research. How do habitual entrepreneurs behave differently? It seems they act to clearly define market opportunities as a matter of priority during venture gestation. What effect does entrepreneurial experience have on future expectations? Clearly a sense of realism is drawn over the difficulties that might be faced, and accords more circumspect judgements of venture survival. This finding informs practitioners considering entrepreneurship for the first time.

Where the actions are : bring reality into virtual reality

This paper describes a work-in-progress on developing design environments that combine wireless and mobile technologies with augmented reality to facilitate bringing context from the physical environment to the virtual models for design work. One of the challenges for designers in a variety of end-user-oriented design disciplines such as architecture and industrial design has been capturing and replaying the contextual information of the intended domain of the artifact being designed. Either the technology is decidedly low-tech, such as charcoal drawings in a sketchbook, out-of-reach, such as immersive virtual reality CAVEs, or a “make-do” with existing technologies, such as a collage of digital photos. This paper describes a novel combination of “off-the-shelf” technologies that may allow designers more capability to create models using standard computer-aided design applications and augmented reality to combine the current, physical context with the projected, digital context. We demonstrate this approach in the building design domain to address a common problem in building construction, construction defect resolution.

Adaptive unsupervised learning of human actions

Automatic detection of suspicious activities in CCTV camera feeds is crucial to the success of video surveillance systems. Such a capability can help transform the dumb CCTV cameras into smart surveillance tools for fighting crime and terror. Learning and classification of basic human actions is a precursor to detecting suspicious activities. Most of the current approaches rely on a non-realistic assumption that a complete dataset of normal human actions is available. This paper presents a different approach to deal with the problem of understanding human actions in video when no prior information is available. This is achieved by working with an incomplete dataset of basic actions which are continuously updated. Initially, all video segments are represented by Bags-Of-Words (BOW) method using only Term Frequency-Inverse Document Frequency (TF-IDF) features. Then, a data-stream clustering algorithm is applied for updating the system's knowledge from the incoming video feeds. Finally, all the actions are classified into different sets. Experiments and comparisons are conducted on the well known Weizmann and KTH datasets to show the efficacy of the proposed approach.

Adaptive and phase transition behavior in performance of discrete multi-articular actions by degenerate neurobiological systems

The identification of attractors is one of the key tasks in studies of neurobiological coordination from a dynamical systems perspective, with a considerable body of literature resulting from this task. However, with regards to typical movement models investigated, the overwhelming majority of actions studied previously belong to the class of continuous, rhythmical movements. In contrast, very few studies have investigated coordination of discrete movements, particularly multi-articular discrete movements. In the present study, we investigated phase transition behavior in a basketball throwing task where participants were instructed to shoot at the basket from different distances. Adopting the ubiquitous scaling paradigm, throwing distance was manipulated as a candidate control parameter. Using a cluster analysis approach, clear phase transitions between different movement patterns were observed in performance of only two of eight participants. The remaining participants used a single movement pattern and varied it according to throwing distance, thereby exhibiting hysteresis effects. Results suggested that, in movement models involving many biomechanical degrees of freedom in degenerate systems, greater movement variation across individuals is available for exploitation. This observation stands in contrast to movement variation typically observed in studies using more constrained bi-manual movement models. This degenerate system behavior provides new insights and poses fresh challenges to the dynamical systems theoretical approach, requiring further research beyond conventional movement models.

Young children’s active citizenship : storytelling, stories, and social actions

This thesis inquires into possibilities for young children‘s active citizenship as provoked through a practice of social justice storytelling with one Preparatory1 class of children aged five to six years. The inquiry was practitioner-research, through a living educational theory approach cultivating an interrelational view of existing with others in evolving processes of creation. Ideas of young children‘s active citizenship were provoked and explored through storytelling, by a storytelling teacher-researcher, a Prep class of children and their teacher. The three major foci of the study were practice, narrative and action. A series of storytelling workshops with a Prep class was the practice that was investigated. Each workshop began with a story that made issues of social justice visible, followed by critical discussion of the story, and small group activities to further explore the story. The focus on narrative was based on the idea of story as a way knowing. Stories were used to explore social justice issues with young children. Metanarratives of children and citizenship were seen to influence possibilities for young children‘s active citizenship. Stories were purposefully shared to provoke and promote young children‘s active citizenship through social actions. It was these actions that were the third focus of the study. Through action research, a social justice storytelling practice and the children‘s responses to the stories were reflected on both in action and after. These reflections informed and shaped storytelling practice. Learning in a practice of social justice storytelling is explained through living theories of social justice storytelling as pedagogy. Data of the children‘s participation in the study were analysed to identify influences and possibilities for young children‘s active citizenship creating a living theory of possibilities for young children‘s active citizenship.

Cluster analysis of movement patterns in multiarticular actions: A tutorial

The present paper proposes a technical analysis method for extracting information about movement patterning in studies of motor control, based on a cluster analysis of movement kinematics. In a tutorial fashion, data from three different experiments are presented to exemplify and validate the technical method. When applied to three different basketball-shooting techniques, the method clearly distinguished between the different patterns. When applied to a cyclical wrist supination-pronation task, the cluster analysis provided the same results as an analysis using the conventional discrete relative phase measure. Finally, when analyzing throwing performance constrained by distance to target, the method grouped movement patterns together according to throwing distance. In conclusion, the proposed technical method provides a valuable tool to improve understanding of coordination and control in different movement models, including multiarticular actions.

The spermostatic and microbicidal actions of quinones and malemides : towards a dual-purpose contraceptive agent

There is an urgent need to develop safe, effective, dual-purpose contraceptive agents that combine the prevention of pregnancy with protection against sexually transmitted diseases. Here we report the identification of a group of compounds that on contact with human spermatozoa induce a state of “spermostasis,” characterized by the extremely rapid inhibition of sperm movement without compromising cell viability. These spermostatic agents were more active and significantly less toxic than the reagent in current clinical use, nonoxynol 9, giving therapeutic indices (ratio of spermostatic to cytotoxic activity) that were orders of magnitude greater than this traditional spermicide. Although certain compounds could trigger reactive oxygen species generation by spermatozoa, this activity was not correlated with spermostasis. Rather, the latter was associated with alkylation of two major sperm tail proteins that were identified as A Kinase-Anchoring Proteins (AKAP3 and AKAP4) by mass spectrometry. As a consequence of disrupted AKAP function, the abilities of cAMP to drive protein kinase A-dependent activities in the sperm tail, such as the activation of SRC and the consequent stimulation of tyrosine phosphorylation, were suppressed. Furthermore, analysis of microbicidal activity using Chlamydia muridarum revealed powerful inhibitory effects at the same low micromolar doses that suppressed sperm movement. In this case, the microbicidal action was associated with alkylation of Major Outer Membrane Protein (MOMP), a major chlamydial membrane protein. Taken together, these results have identified for the first time a novel set of cellular targets and chemical principles capable of providing simultaneous defense against both fertility and the spread of sexually transmitted disease.