Basis For Action Research Articles

Noninvasive Human Brain Stimulation

Noninvasive brain stimulation with transcranial magnetic stimulation (TMS) or transcranial direct current stimulation (tDCS) is valuable in research and has potential therapeutic applications in cognitive neuroscience, neurophysiology, psychiatry, and neurology. TMS allows neurostimulation and neuromodulation, while tDCS is a purely neuromodulatory application. TMS and tDCS allow diagnostic and interventional neurophysiology applications, and focal neuropharmacology delivery. However, the physics and basic mechanisms of action remain incompletely explored. Following an overview of the history and current applications of noninvasive brain stimulation, we review stimulation device design principles, the electromagnetic and physical foundations of the techniques, and the current knowledge about the electrophysiologic basis of the effects. Finally, we discuss potential biomedical and electrical engineering developments that could lead to more effective stimulation devices, better suited for the specific applications.

Effect of extracorporeal shock wave therapy on the biochemical composition and metabolic activity of tenocytes in normal tendinous structures in ponies

Extracorporeal shockwave therapy (ESWT) has recently been introduced as a new therapy for tendon injuries in horses, but little is known about the basic mechanism of action of this therapy. To study the effect of ESWT on biochemical parameters and tenocyte metabolism of normal tendinous structures in ponies. Six Shetland ponies, free of lameness and with ultrasonographically normal flexor and extensor tendons and suspensory ligaments (SL), were used. ESWT was applied at the origin of the suspensory ligament and the mid-metacarpal region of the superficial digital flexor tendon (SDFT) 6 weeks prior to sample taking, and at the mid-metacarpal region (ET) and the insertion on the extensor process of the distal phalanx (EP) of the common digital extensor tendon 3 h prior to tendon sampling. In all animals one front leg was treated and the other front leg was used as control. After euthanasia, tendon explants were harvested aseptically for in vitro cell culture experiments and additional samples were taken for biochemical analyses. In the explants harvested 3 h after treatment, glycosaminoglycan (GAG) and protein syntheses were increased (P<0.05). The synthesis of all measured parameters was decreased 6 weeks after ESWT treatment. Biochemically, the level of degraded collagen was increased 3 h after treatment (P<0.05). Six weeks after treatment, there was a decrease of degraded collagen and GAG contents. DNA content had not changed in either tendon samples or explants after culturing. ESWT causes a transient stimulation of metabolism in tendinous structures of ponies shortly after treatment. After 6 weeks metabolism has decreased significantly and GAG levels are lower than in untreated control limbs. The stimulating short-term effect of ESWT might accelerate the initiation of the healing process in injured tendons. The long-term effect seems less beneficial. Further research should aim at determining the duration of this effect and at assessing its relevance for end-stage tendon quality.

Human reward system activation is modulated by a single dose of olanzapine in healthy subjects in an event-related, double-blind, placebo-controlled fMRI study

Olanzapine is a neuroleptic drug widely prescribed to treat schizophrenia and bipolar disorder. Although it is long known that modulation of the dopamine system is a basic mechanism of action of neuroleptics, their impact on reward functions mediated by dopamine is still poorly understood. Using functional magnetic resonance imaging (fMRI), we intended to reveal the effects of a single dose of olanzapine on reward-related brain activation. Eight healthy subjects were each scanned twice, once 5 h after intake of 5 mg of olanzapine and once after intake of placebo in a double-blind cross-over design. Subjects performed a delayed incentive paradigm with monetary reward to investigate reward functions and a breath-holding task as a hypercapnic challenge to reveal unspecific drug effects on the fMRI signal. Reward-related brain activation in the ventral striatum, anterior cingulate and inferior frontal cortex was reduced on olanzapine compared to placebo. Only the differential effects (high>no reward) in the ventral striatum were independent of overall drug effects as measured with the breath-holding task. Parallel to the differential effects in the ventral striatum, the acceleration of reaction times in the trials with higher rewards was diminished in the olanzapine sessions. Our behavioural and fMRI results can be interpreted as first evidence from neuroimaging that olanzapine affects the assignment of incentive salience represented by differential activation in dopaminergic brain areas and acceleration of reaction times. This can help to better understand neuroleptic effects in psychiatric diseases. Furthermore, we demonstrate the value of a hypercapnic challenge in functional pharmaco-MRI.

Management as a Design Science Mindful of Art and Surprise

Huff: Management is a relatively young profession and a young field of study. For some time, there has been an interest in as a primary descriptor of management practice. Herbert Simon described management as a science in The Sciences of the Artificial, first published in 1969 and reissued in a third edition in 1996, yet the specifics of design have not been clear to me, especially as the basis for action. My interest in design was recently piqued by David Tranfield, professor of management at Cranfield School of Management, who then introduced me to the work of Joan (pronounced Johan for those of you not familiar with names from the Netherlands) van Aken, professor of organization and management at Eindhoven University of Technology. van Aken’s work was very interesting, but I wasn’t initially convinced that management conceived as design could incorporate two metaphors for management that I have been more inclined to use: an artful blend of resources and action and the to respond to surprise. The following conversation with David and Joan provided a convincing positive answer. We discuss the definition of design, its importance, the art of design, its capacity to deal with surprise, inevitable limitations, how theory is involved, and a vision of the future. I hope that readers will find the basic idea as significant as I do.

Immunonutrition in septic patients: A philosophical view of the current situation

Two different ways of thinking pervaded the history of science: rationalism and empiricism. In theory, these two paradigms are not necessarily in conflict. In practice, there has always been tension between them. The coming of evidence-based medicine put empiricism in a privileged position, but empiricism without a rationalistic guide could even be usefulness. The aim of this work is to present the tension between the rational reasons to administer immunonutrients to patients with sepsis and the controversial empirical evidence stemming from clinical trials. We reviewed the literature on immunonutrition in sepsis from the rationalist and the empiricist perspectives. The large body of evidence for positive effects of immunonutrients in experimental models and the contradictory results from clinical trials make the discussion on immunonutrition in sepsis a typical example where the conflict between rationalism and empiricism hampered the advancement of knowledge and the implementation of new effective therapies into clinical practice. Future research projects involving immunonutrients should be based on robust knowledge of basic mechanisms of action to be properly addressed in clinical trials.

Evidence supporting the hypothesis that one of the main functions of the aryl hydrocarbon receptor is mediation of cell stress responses

We have previously proposed that one of the major consequences of activation of the aryl hydrocarbon receptor (AhR) by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) could be elicitation of 'cell stress response' reactions [Matsumura, Biochem. Pharmacol. 66 (2003), 527-540]. This hypothesis was based mainly on the similarity between the toxic symptoms, particularly those related to the wasting syndrome, and those induced by bacterial endotoxins, namely lipopolysaccharides (LPS) in vivo, as well as the biochemical and molecular consequences of their toxic actions in vitro. Since the basic action mechanism of LPS as an inducer of cell stress responses (CSR) is known to some extent, including knowledge of their specific receptors (i.e., toll-like receptors) and their signaling process through the inflammatory response messengers, the above comparison offered a good point of reference to this subject. Furthermore, the process of constructing this hypothesis itself has provided us with a good opportunity to give a fresh view on the toxic action patterns of TCDD.

Antiangiogenesis and Drug Delivery to Tumors: Bench to Bedside and Back

After over 30 years of preclinical and clinical development, antiangiogenic agents have recently entered the clinic as attractive targeted therapeutics for the treatment of cancer. Fueled by exciting new developments in the field, the AACR Special Conference was designed to broadly survey critical scientific advances in the antiangiogenic therapy of cancer. Because these advances have come primarily with the use of combinations of antiangiogenic agents with chemotherapy, or with antiangiogenic agents that also directly target the cancer cells, the central theme included the issue of drug delivery to tumors. These two major issues were addressed in concert, from basic mechanisms of action of antiangiogenic agents to new combination approaches to cancer treatment. Nearly 300 participants from 20 countries registered for the conference, drawn both from academia and industry, with a wide range in experience and background. Dr. Rakesh Jain, along with conference co-chairs, Drs. Lee Ellis and Luisa Iruela-Arispe, assembled an outstanding lineup of speakers for this conference that included many of the pioneers in the fields of angiogenesis and drug delivery from the U.S. and abroad. This resulted in an excellent overview of the advances in our understanding of cellular and molecular aspects of tumor angiogenesis and antiangiogenic therapy of cancer in combination with conventional therapy.

The effect of different anticoagulants on thrombin generation

Decrease in thrombin generation is the key effect in anticoagulation. The aim of the present study was to investigate the effect of anticoagulants on thrombin generation and the relation to platelet count. Plasma samples from 10 healthy volunteers (mean age 43.0 +/- 9 years) were incubated at preset platelet counts with different doses of the anticoagulants lepirudin, fondaparinux and low molecular weight heparins. Thrombin generation was measured in a tissue factor-mediated assay using a fluorometer and a slow-reacting fluorogenic substrate. The endogenous thrombin potential, the lag phase, the maximum reaction velocity (Vmax) and the concentration of a given anticoagulant required for 50% inhibition of thrombin generation (IC50) are presented. All three anticoagulants decreased endogenous thrombin potential and prolonged the lag phase in a dose-dependent manner. Fondaparinux and low molecular weight heparins, but not hirudin, decreased Vmax in a concentration-dependent manner. With increasing platelet count, the IC50 increased but the extent of this increase was not uniform for the three anticoagulants and the three variables investigated. The influence of anticoagulants on thrombin generation is variable, depending on their basic mechanism of action. In defining and comparing their effects, the endogenous thrombin potential, the lag phase and the maximum reaction velocity should be considered together. Platelets have a considerable influence on the magnitude of thrombin generation.

Veterinary Radiation Therapy: Review and Current State of the Art

Veterinary radiation oncology became a recognized specialty in 1994. Radiation therapy is an important component of a multimodality approach to treating cancer in companion animals. It is important to understand the many aspects of radiation, including the equipment used in external beam radiation therapy, the basic mechanism of action of ionizing radiation, the results of irradiating various histopathological types of tumors, as well as the associated potential acute and late side effects of radiation. A comprehensive review of radiation therapy is timely and provides information for clients on cancers that may benefit from external beam radiation therapy.

Antimicrobial Peptides: Basic Mechanisms of Action and Emerging Pharmacological Interest

Antimicrobial Peptides: Basic Mechanisms of Action and Emerging Pharmacological Interest

Adventures with ABC-proteins: Highly Conserved ATP-dependent Transporters

The general properties of ABC transporters, from bacteria to humans, including a brief history of their initial discovery, are considered. ABC transporters, one of the largest protein super families and vital for human health, are in toto responsible for the transport of an enormous range of molecules from ions (CFTR) or anti-tumour drugs (Pgp/MDR) to large polypeptides. Nevertheless, all ABC transporters are powered by a conserved ATPase the ABC or NBD domain, using in all probability the same basic mechanism of action for the hydrolysis of ATP and its coupling to the transport process. Based on recent high resolution structures of several NBDs and an intact transporter, a model of how dimers of these important proteins function will be discussed, with particular attention to HlyB, the ABC transporter from E. coli.

Role of nuclear receptors in lung tumourigenesis

Nuclear receptors are a family of ligand dependent transcription factors which have important roles in control of growth and differentiation in many cell types. This review will focus on the role of two members of this family (estrogen receptors and PPARγ) in the initiation and progression of lung cancer. These receptors have been studied in other types of cancer, and the differences between findings in lung cancer and other malignancies will be discussed. We will also describe recent studies on the basic mechanisms of action of these receptors and suggest novel therapeutic targets. Specifically, there is emerging evidence that these receptors can induce gene expression through both ligand-dependent and ligand-independent pathways, and distinct families of genes are likely to be regulated depending on the mechanism of nuclear receptor signalling. These data suggest that a greater understanding of the basic biology and mechanisms of these receptors is required to develop specific pharmacological agents as therapeutics for cancer. Specifically, the development of selective agonists for these receptors, such as tamoxifen, may lead to more selective engagement of anti-tumourigenic pathways. In addition, combinatorial therapy using selective nuclear receptor activators in conjunction with the recently developed EGF receptor inhibitors (gefitinib, erlotinib) may sensitise cells which are unresponsive to the effects of EGF receptor inhibitors alone, providing powerful new therapeutic strategies.

Current and Potential Uses of Imiquimod

Imiquimod, an imidazoquinoline amine, is an immune response modifier first FDA-approved for the treatment of external genital and perianal warts in 1997. Since its appearance on the market, its antiviral and antitumor properties have been used in the treatment of a variety of dermatologic conditions. In this review article, the basic mechanism of action of imiquimod, current FDA-approved and non-FDA-approved uses of imiquimod, and key points of medication application frequency, possible adverse effects, and use in combination therapy are discussed. Common skin conditions that may be eradicated with imiquimod are emphasized.

Airway Biofilms

The differentiation of bacterial biofilms in the airway environment, the pathogenesis of airway biofilm, and possible therapeutic methods are discussed. Biofilm diseases that characteristically involve the respiratory system include cystic fibrosis (CF), diffuse panbronchiolitis (DPB), and bronchiectasia with Pseudomonas aeruginosa (P. aeruginosa) infection. There is evidence to suggest that almost all strains of P. aeruginosa have the genetic capacity to synthesize alginate, a main matrix of biofilms, when ecological conditions are unfavorable for their survival. The bacteria inside the mature biofilm show increased resistance to both antibacterials and phagocytic cells, express fewer virulence factors because of their stationary state of growth, and are less stimulatory to the mucosa because of the 'sandwich binding'. These factors facilitate both the colonization of bacteria and their extended survival even under unfavorable conditions. Since the biofilm limits colonization to a latent form, the clinical symptoms in this situation are unremarkable. However, the clinical progression of both CF and DPB proceeds in two characteristic directions. The first is an acute exacerbation caused by planktonic bacteria that have germinated from the biofilm. The second is a slow progression of disease that is induced by harmful immune reactions. The harmful reactions are mediated by alginate, which induces antigen antibody reactions around the airways, as well as formation of circulating immune complexes that are deposited on lung tissue. Furthermore, the highest titer of bacterial permeability increasing anti-neutrophil cytoplasmic autoantibodies (BPI-ANCA) is observed in association with highly impaired pulmonary function in patients with CF and DPB, as well as in patients with a lengthy period of colonization with P. aeruginosa. BPI-ANCA subsequently makes chronic airway infection even more intractable. The long-term use of 14- or 15-ring membered macrolides results in a favorable clinical outcome for patients with DPB and in some patients with CF. In the last 10 years, an increasing number of studies have reported secondary actions of macrolides that include effects on both airway and phagocytic cells, as well as an anti-biofilm activity. The 14- or 15-ring membered macrolides inhibit: (i) the alginate production from P. aeruginosa; (ii) the antibody reaction to alginate, which leads to a decrease in the immune complex formation; and (iii) the activation of the autoinducer 3-O-C12-homoserine lactone and subsequent expression of lasI and rhlI in quorum sensing systems in P. aeruginosa. These anti-biofilm actions of macrolides may represent their basic mechanisms of action on airway biofilm disease.

Distinct effects of sleep deprivation on binding to norepinephrine and serotonin transporters in rat brain

There is evidence to suggest that the antidepressant activity of sleep deprivation may be due to an enhancement of serotonergic and/or noradrenergic neurotransmission in brain. In the present study we examined the possibility that such changes may occur at the level of the norepinephrine (NET) and serotonin (SERT) and transporters. Rats were deprived of sleep for 96 h using the modified multiple platform method and then sacrificed for autoradiographic assessments of NET and SERT binding throughout the brain. [ 3H]Nisoxetine binding to the NE transporter was generally decreased in 44 of 45 areas examined, with significant reductions occurring in the anterior cingulate cortex (−16%), endopiriform n. (−18%), anterior olfactory n. (−19%), glomerular layer of olfactory bulb (−18%), ventral pallidum (−14%), medial preoptic area (−16%), retrochiasmatic/arcuate hypothalamus (−18%), anteromedial thalamic n. (−15%), and rostral raphe (−17%). In contrast, SERT binding measured with [ 11C]DASB showed no clear directional trends in 61 brain areas examined, but was significantly reduced in subdivisions of the anterior olfactory nucleus (−22%) and substantia nigra (−18%). Thus, sleep deprivation induced widespread decreases in NET binding, and fewer and well-localized decreases in SERT binding. Significant down-regulation in one brain region, the anterior olfactory nucleus, was observed in the case of both transporters. These results suggest that mechanisms involved in the antidepressant action of sleep deprivation may involve generalized NET down-regulation as well as decreased SERT binding in specific areas. Insofar as these changes may be associated with increased levels of serotonin (5-HT) and norepinephrine (NE) in the synapse, they suggest that sleep deprivation may share some basic mechanisms of action with several current antidepressant medications.

The Drosha-DGCR8 complex in primary microRNA processing

RNase III proteins play key roles in microRNA (miRNA) biogenesis. The nuclear RNase III Drosha cleaves primary miRNAs (pri-miRNAs) to release hairpin-shaped pre-miRNAs that are subsequently cut by the cytoplasmic RNase III Dicer to generate mature miRNAs. While Dicer (class III) and other simple RNase III proteins (class I) have been studied intensively, the class II enzyme Drosha remains to be characterized. Here we dissected the action mechanism of human Drosha by generating mutants and by characterizing its new interacting partner, DGCR8. The basic action mechanism of Drosha was found to be similar to that of human Dicer; the RNase III domains A and B form an intramolecular dimer and cleave the 3' and 5' strands of the stem, respectively. Human Drosha fractionates at approximately 650 kDa, indicating that Drosha functions as a large complex. In this complex, Drosha interacts with DGCR8, which contains two double-stranded RNA (dsRNA)-binding domains. By RNAi and biochemical reconstitution, we show that DGCR8 may be an essential component of the pri-miRNA processing complex, along with Drosha. Based on these results, we propose a model for the action mechanism of class II RNase III proteins.

Organization of Ca 2+ Release Units in Excitable Smooth Muscle of the Guinea-Pig Urinary Bladder

Ca 2+ release from internal stores (sarcoplasmic reticulum or SR) in smooth muscles is initiated either via pharmaco-mechanical coupling due to the action of an agonist and involving IP3 receptors, or via excitation-contraction coupling, mostly involving L-type calcium channels in the plasmalemma (DHPRs), and ryanodine receptors (RyRs), or Ca 2+ release channels of the SR. This work focuses attention on the structural basis for the coupling between DHPRs and RyRs in phasic smooth muscle cells of the guinea-pig urinary bladder. Immunolabeling shows that two proteins of the SR: calsequestrin and the RyR, and one protein the plasmalemma, the L-type channel or DHPR, are colocalized with each other within numerous, peripherally located sites located within the caveolar domains. Electron microscopy images from thin sections and freeze-fracture replicas identify feet in small peripherally located SR vesicles containing calsequestrin and distinctive large particles clustered within small membrane areas. Both feet and particle clusters are located within caveolar domains. Correspondence between the location of feet and particle clusters and of RyR- and DHPR-positive foci allows the conclusion that calsequestrin, RyRs, and L-type Ca 2+ channels are associated with peripheral couplings, or Ca 2+ release units, constituting the key machinery involved in excitation-contraction coupling. Structural analogies between smooth and cardiac muscle excitation-contraction coupling complexes suggest a common basic mechanism of action.

Production of recombinant C5a from rainbow trout ( Oncorhynchus mykiss): role in leucocyte chemotaxis and respiratory burst

Activation of the complement system can lead to the formation of the membrane attack complex, in which the component C5 is cleaved into C5a and C5b fragments. The C5a anaphylatoxin is a very potent pro-inflammatory molecule that induces chemotaxis and respiratory burst processes in a variety of mammalian leucocytes. While C5a has been well studied in mammals, little is known about the structure and function of C5a in teleost fish or other non-mammalian species. In the present study, we have produced and purified recombinant rainbow trout C5a (rtC5a), and we have shown that it plays an important role in inducing leucocyte migration as well as in triggering the respiratory burst of peripheral blood (PBLs) and head kidney leucocytes (HKLs). When the carboxy-terminal Arg was removed from rtC5a, its ability to induce cell migration and superoxide production remained intact. Interestingly, we show that leucocytes migrating towards rtC5a attached to the plate with a well-spread circular morphology, whereas those migrating towards activated trout serum displayed more irregular and dendritic-like shapes. Our data suggest that the basic mechanisms of action of the C5a anaphylotoxin have remained conserved for more than 300 million years.

Environmental estrogens can affect the function of mussel hemocytes through rapid modulation of kinase pathways

Estrogens and estrogenic chemicals can affect several vertebrate non-reproductive functions, the immune response in particular. We have previously shown that in the hemocytes of the marine mollusc Mytilus the natural estrogen 17β-estradiol (E 2) can affect the immune function through rapid tyrosine kinase-mediated signalling pathways converging on phosphorylation of both mitogen activated protein kinases (MAPKs) and signal transducers and activators of transcription (STATs), whose activation plays a key role in the immune response. In this work the effects of synthetic estrogens (such as DES), estrogenic chemicals (such as Bisphenol A, Nonylphenol), and plant estrogens (genistein) on mussel hemocytes were evaluated. The results demonstrate that all the EDCs tested exert in vitro effects similar to those of E 2 on lysosomal membrane stability, although at concentrations 1000 times higher than those of the natural estrogen. When the effects of DES, BPA, and NP on tyrosine kinase-mediated cell signalling were investigated, estrogenic compounds showed distinct effects on the phosphorylation state of MAPK and STAT members. In particular, only DES, like E 2, induced p38 MAPK phosphorylation, whereas BPA and NP seem to have opposite effects. Moreover, different EDCs significantly decreased the tyrosine phosphorylation state of STAT3 and STAT5, showing a distinct effect with respect to E 2. Experiments with specific kinase inhibitors showed that activation of p38 MAPK, but also of ERK MAPK and PI3-kinase, plays a key role in mediating the effect of DES. On the other hand, the effects of NP were partly mediated by ERK MAPK activation. BPA-induced lysosomal membrane destabilisation was unaffected by either MAPK or PI3-K inhibitors. However, hemocyte pre-treatment with the PKC inhibitor GF109203X prevented the effects of both BPA and NP, this indicating that kinase pathways other than those involving MAPKs are also responsible for mediating the effects of certain EDCs. Overall, the results support the hypothesis that EDCs may rapidly modulate the function of mussel hemocytes through activation of transduction pathways involving different kinase-mediated cascades. Moreover, the effects of EDCs on the phosphorylation state of transcription factor STATs suggest that these compounds may lead to changes in gene expression secondary to modulation of kinase/phosphatases. Our data address to the importance of investigating full range responses to estrogenic chemicals and may help understanding their basic mechanisms of action in ecologically relevant invertebrate species.

A Rational Use of Immune Enhancing Diets: When Should We Use Dietary Arginine Supplementation?

Controversies in any arena of human activity often result in the polarization of the individuals involved into 2 opposing camps. Controversy about the use of immune-enhancing diets (IEDs) is no exception. On one hand, some groups are proposing indiscriminate use of IEDs, whereas others have created guidelines advocating that their use should be banned for the critically ill. At stake is an emerging paradigm: that dietary manipulation of the immune system is possible and may become an important adjunct to other therapies, thus helping prevent or treat multiple diseases for millions worldwide. Under these circumstances, extremist claims of miraculous benefits or inappropriate assertions of evil can only delay the emergence of a nascent science. This paper is therefore a plea for moderation from both camps, lest we cause irreparable damage to our clinical practices and potential injury to individual patients. IEDs all contain arginine. However, they also contain other substances such as omega-3 fatty acids, and nucleotides. The use of all these nutrients together into commercial IEDs without adequate evaluation of their individual effects has prevented the development of mechanistic hypotheses of action. Despite this, IEDs have been tested extensively, allowing the development of guidelines for their use. IEDs should be used for surgical patients, especially those undergoing elective surgery. IEDs show no benefit and indeed can potentially harm patients with sepsis, especially in the nonsurgical group, and should not be used outside of research protocols. Advances in basic research have helped us understand mechanisms of how arginine contained in IEDs may help surgical patients but may be deleterious in patients with sepsis. A review of the basic mechanisms of action of arginine on the immune system is enclosed in this paper and should serve as a basis for the development of scientific principles that guide clinical use of IEDs.