Unexpected Action Research Articles

Plant HDAC inhibitor chrysin arrest cell growth and induce p21 WAF1 by altering chromatin of STAT response element in A375 cells

BackgroundChrysin and its analogues, belongs to flavonoid family and possess potential anti-tumour activity. The aim of this study is to determine the molecular mechanism by which chrysin controls cell growth and induce apoptosis in A375 cells.MethodsEffect of chrysin and its analogues on cell viability and cell cycle analysis was determined by MTT assay and flowcytometry. A series of Western blots was performed to determine the effect of chrysin on important cell cycle regulatory proteins (Cdk2, cyclin D1, p53, p21, p27). The fluorimetry and calorimetry based assays was conducted for characterization of chrysin as HDAC inhibitor. The changes in histone tail modification such as acetylation and methylation was studied after chrysin treatment was estimated by immuno-fluorescence and western blot analysis. The expression of Bcl-xL, survivin and caspase-3 was estimated in chrysin treated cells. The effect of chrysin on p21 promoter activity was studied by luciferase and ChIP assays.ResultsChrysin cause G1 cell cycle arrest and found to inhibit HDAC-2 and HDAC-8. Chrysin treated cells have shown increase in the levels of H3acK14, H4acK12, H4acK16 and decrease in H3me2K9 methylation. The p21 induction by chrysin treatment was found to be independent of p53 status. The chromatin remodelling at p21WAF1 promoter induces p21 activity, increased STAT-1 expression and epigenetic modifications that are responsible for ultimate cell cycle arrest and apoptosis.ConclusionChrysin shows in vitro anti-cancer activity that is correlated with induction of histone hyperacetylation and possible recruitment of STAT-1, 3, 5 proteins at STAT (−692 to −684) region of p21 promoter. Our results also support an unexpected action of chrysin on the chromatin organization of p21WAF1 promoter through histone methylation and hyper-acetylation. It proposes previously unknown sequence specific chromatin modulations in the STAT responsive elements for regulating cell cycle progression negatively via the induction of the CDK inhibitor p21WAF1.

Regulatory mobilization and service delivery at the edge of the regulatory state

AbstractWhere there is weak state capacity to carry out regulatory, redistributional, and developmental functions characterizing much of the developing world, the role of governance and service delivery is also performed by a myriad of private actors. Institutional reform in the utility sector in developing countries has often failed to distinguish between social and economic regulation. I show how private actors like NGOs and local community groups undertake what I term “regulatory mobilization” to influence the new rules of the service delivery game, as well as to deliver much‐needed basic services to urban poor communities. Based on extensive fieldwork carried out in the Philippines, this article reveals and explains the politics of the informal sector at the edge of the regulatory state. More than a decade since the privatization of the Metropolitan Waterworks and Sewerage System in Metro Manila in 1997, water access for the urban poor remained limited as privatized water utilities faced difficulties in extending service provision. In the context of an unpredictable regulatory landscape and an oligarchic patrimonial state, unexpected collective action by organized urban poor communities and NGOs has taken place around water as a subsistence right. Combining hybrid mobilizations to obtain water as well as influencing the rules governing their provision, these forms of regulatory mobilization appear to be peripheral and episodic. However, depending on how local and sectoral politics are conflated, such regulatory mobilization may sometimes not only result in obtaining subsistence goods, but may also occasionally project countervailing power in the policy sector, and influence formal regulatory frameworks in surprising ways.

Increased orienting to unexpected action outcomes in schizophrenia

Although some recent research has indicated reduced performance monitoring in patients with schizophrenia, the literature on this topic shows some remarkable inconsistencies. While most studies suggest diminished error signals following error responses, some studies reported normal post-error slowing, while others reported reduced post-error slowing. Here we review these studies and highlight the most important discrepancies. Furthermore, we argue that overall error rates are a mostly neglected issue that can at least partly explain these discrepancies. It has been reported previously that post-error slowing depends on the error rates. Participants or patients that make more errors are likely to show decreased post-error slowing. Therefore, when a group of patients is compared to a group of controls, it is extremely important to match error rates. For this purpose, we developed a procedure where we matched individuals' error rates. In a task where subjects had to press a response key corresponding to one of four colors we manipulated the difficulty on an individual basis by varying the discriminability between the colors. Schizophrenic patients and a group of controls were tested with this procedure showing that differences in accuracy disappear. Interestingly, we can see that in patients, the color values that were needed to reach similar levels of accuracy correlate with the Positive and Negative Syndrome Scale (PANSS) scale, with higher PANSS requiring more color. Most important, we found that schizophrenic patients have increased rather than decreased post-error slowing when the inter-trial interval (ITI) is short. This result can be interpreted within the framework of the orienting account, as it has been demonstrated previously that schizophrenic patients show increased distractibility.

“Stricken to Silence”: Authoritative Response, Homeric Irony, and the Peril of a Missed Language Cue

The formula2 Thus he spoke, but they in fact all were stricken to (translation ommitted)3 occurs sixteen times in Homer4 and has received significant treatment in a number of recent studies focusing on its referential force. Its connotative level of signification (Kelly 2007:6) has been projected in part for the Iliad, and important themes and functions have been suggested. Silvia Montiglio (1993:175-78) has considered the formula's meaning within the Iliad both etymologically and more generally, and found that it suggests une rupture anormale, la dechirure of the normal communication process. John Miles Foley has linked the formula in the Iliad with the speech that precedes it, since each initial speech proposes or reports a radical, usually unexpected action (1995:13) that promises either the winning or losing of kleos. Foley's research further demonstrates that the formula leads, immediately or inevitably, to the qualification if not dismissal of the proposed or reported action (15) that precedes the silence formula. Raymond Person (1995) uses conversation analysis to suggest that the formula marks that a speaker will follow with a dispreferred response, essentially a response that is delayed and mitigated. Adrian Kelly's study (2007:85-86) of the formula in the Iliad highlights the relationship between the speech that immediately precedes the formula and the speech that ensues, in terms of agreement or disagreement.5

Phospholipase C-ε links Epac2 activation to the potentiation of glucose-stimulated insulin secretion from mouse islets of Langerhans

Glucose-stimulated insulin secretion (GSIS) from pancreatic β-cells is potentiated by cAMP-elevating agents, such as the incretin hormone glucagon-like peptide-1 (GLP-1), and cAMP exerts its insulin secretagogue action by activating both protein kinase A (PKA) and the cAMP-regulated guanine nucleotide exchange factor designated as Epac2. Although prior studies of mouse islets demonstrated that Epac2 acts via Rap1 GTPase to potentiate GSIS, it is not understood which downstream targets of Rap1 promote the exocytosis of insulin. Here, we measured insulin secretion stimulated by a cAMP analog that is a selective activator of Epac proteins in order to demonstrate that a Rap1-regulated phospholipase C-epsilon (PLC-ε) links Epac2 activation to the potentiation of GSIS. Our analysis demonstrates that the Epac activator 8-pCPT-2’-O-Me-cAMP-AM potentiates GSIS from the islets of wild-type (WT) mice, whereas it has a greatly reduced insulin secretagogue action in the islets of Epac2 (-/-) and PLC-ε (-/-) knockout (KO) mice. Importantly, the insulin secretagogue action of 8-pCPT-2’-O-Me-cAMP-AM in WT mouse islets cannot be explained by an unexpected action of this cAMP analog to activate PKA, as verified through the use of a FRET-based A-kinase activity reporter (AKAR3) that reports PKA activation. Since the KO of PLC-ε disrupts the ability of 8-pCPT-2’-O-Me-cAMP-AM to potentiate GSIS, while also disrupting its ability to stimulate an increase of β-cell [Ca2+]i, the available evidence indicates that it is a Rap1-regulated PLC-ε that links Epac2 activation to Ca2+-dependent exocytosis of insulin.

On Monetary Policy and Stock Market Anomalies

This study utilizes a macro-based VAR framework to investigate whether stock portfolios formed on the basis of their value, size and past performance characteristics are affected in a differential manner by unexpected US monetary policy actions during the period 1967-2007. Full sample results show that value, small capitalization and past loser stocks are more exposed to monetary policy shocks in comparison to growth, big capitalization and past winner stocks. Subsample analysis, motivated by variation in the realized premia and parameter instability, reveals that monetary policy shocks’ impact on these portfolios is significant and pronounced only during the pre-1983 period.

Great Unexpectations

A sermon by Bill Gaventa, reflecting on the ways that occasional unusual and unexpected actions by people with disabilities challenge our preconceptions and expectations based on the label of disability, and illustrate the Biblical ways in which God is revealed in unexpected people, in unexpected places and unexpected ways.

Orienting towards unexpected action outcomes

Event Abstract Back to Event Orienting towards unexpected action outcomes Wim Notebaert1* 1 Experimental Psychology, Ghent University, Belgium Recently we proposed the orienting account for post-error slowing, stating that RT slowing after errors is related to attentional capture by infrequent errors. In this presentation I will discuss several new studies that have confirmed various predictions of this account. We have demonstrated that the P3 (ERP component related to orienting) predicts post-error slowing, that schizophrenic patients (known to be more distractible) show increased post-error slowing compared to controls when the intertrial interval is short, and that post-error slowing is an a-specific effect that transfers over tasks. We also investigated post-error slowing in an experiment without external feedback. We showed that post-error slowing also increases as the likelihood of an error decreases, suggesting that orienting to unexpected action outcomes does not require external feedback. Finally, when manipulating the saliency of an error, we demonstrated more post-error slowing after more salient errors. Importantly, while these empirical data were predicted by the orienting account, more traditional strategic accounts for error monitoring have difficulties in explaining these observations. Keywords: Attention, P3 Conference: XI International Conference on Cognitive Neuroscience (ICON XI), Palma, Mallorca, Spain, 25 Sep - 29 Sep, 2011. Presentation Type: Symposium: Oral Presentation Topic: Symposium 23: Regulating access to consciousness: Cortical mechanisms of attention Citation: Notebaert W (2011). Orienting towards unexpected action outcomes. Conference Abstract: XI International Conference on Cognitive Neuroscience (ICON XI). doi: 10.3389/conf.fnhum.2011.207.00602 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 14 Nov 2011; Published Online: 28 Nov 2011. * Correspondence: Prof. Wim Notebaert, Experimental Psychology, Ghent University, Ghent, Belgium, wim.notebaert@ugent.be Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Wim Notebaert Google Wim Notebaert Google Scholar Wim Notebaert PubMed Wim Notebaert Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

ChemInform Abstract: The Discovery of Antidepressants: A Winding Path

Modern treatment of mental depression started with the availability of monoamine oxidase (MAO) inhibitors and tricyclic antidepressants. These drugs also contributed to the early development of psychopharmacology. Attempts to improve the anti-tuberculous action of the hydrazine derivative isoniazid by developing derivatives thereof led to the synthesis of iproniazid. Its introduction as the first modern antidepressant was based on three unexpected actions of the drug: MAO-inhibition, ‘reversal’ of reserpine-induced sedation, and the presence of psychostimulation as a clinical side effect in man. However, the initial success of iproniazid and other MAO inhibitors, hydrazides and non-hydrazides, was curtailed by the occurrence of undesirable side effects such as potentiation of the blood-pressure elevating action of food amines. The tricyclic antidepressants were a development of the class of antihistamines, one of which, chlorpromazine, showed neuroleptic activity. A congener of this compound, imipramine, was discovered by clinical observation to have unexpected antidepressant effects. The clinical success of this drug (which is still in use) led to the development of a successful series of other tricyclic and non-tricyclic antidepressants. Progress in the elucidation of possible mechanisms of the action of the tricyclic compounds has helped this development. Recent advances in basic research have also induced a revival of MAO-inhibitors since, due to the discovery of MAO-subtypes, inhibitors with higher specificity and fewer undesirable side effects are now available.

Serum undercarboxylated osteocalcin was inversely associated with plasma glucose level and fat mass in type 2 diabetes mellitus

Dear Editor, We read with interest the article by Kanazawa and coauthors [1] about osteocalcin (OC) and its metabolic associations in a cross-sectional study of a Japanese population affected by type 2 diabetes mellitus. The authors suggest that undercarboxylated OC (ucOC) is associated with fat mass, especially visceral fat, as well as with glucose level in diabetic men, claiming that their study seems to be the first clinical one suggesting that not only OC but also ucOC is associated with lipid/glucose metabolism in type 2 diabetes. According to the authors, the association showed in their population between OC or ucOC versus glucose level seems not to be because of OC-modulated insulin sensitivity or secretion. Several studies, in fact, have indicated that hyperglycemia can induce a low turnover of bone by evoking osteoblast dysfunction and suppressing serum OC level, as depicted by alterations of bone markers before and after treatment of diabetes [2, 3]. So they hypothesize that, due to the lack of correlation between OC or ucOC versus fasting C peptide as surrogate marker for endogenous insulin secretion in their diabetic patients, it would be high glucose level which suppresses osteoblast function and, consequently, OC expression. Thus, according to contrasting reports in scientific literature, which showed how Esp mice, having an excess of ucOC, display severe hypoglycemia and protection against dietinduced obesity and diabetes [4], the cause and effect between glicemic control and OC is still unresolved. Adipose tissue (AT) is nowadays considered a true endocrine organ and a source both of AT-specific cytokines (adipokines) and inflammatory cytokines. We have recently reported that human AT expresses both OC and all genes involved in OC carboxylation and, in general, in Glaprotein carboxylation, both producing and releasing OC protein [5]. So the metabolic effects of OC could be partially mediated by an organ striktly implicated in insulinresistant conditions, due to its endocrine and metabolic functions and its direct cross-talking with the liver through the portal vein. The authors described also a negative correlation between ucOC and the fat mass [1] and our findings, showing OC production by AT [5], could partially explain it. The balance between ucOCN and cOCN, in fact, could play a role in the pathophysiology of metabolic diseases, as depicted by studies both in Esp hypoglicemic rats and in obese men [4, 5]. AT, being able to release both forms, could be again one of metabolism actors if we consider that, compared with bone, AT is highly represented in the human body and can act both in an endocrine and paracrine way. On the basis of our results, revealing unexpected actions of AT, we suggest a different approach in order to elucidate the association between diabetes, bone osteoporosis, and cardiovascular diseases, and the role of two different cell types sharing a common precursors, the adipocyte and the osteoblast.

Theta-activity in anterior cingulate cortex predicts task rules and their adjustments following errors

Accomplishing even simple tasks depend on neuronal circuits to configure how incoming sensory stimuli map onto responses. Controlling these stimulus-response (SR) mapping rules relies on a cognitive control network comprising the anterior cingulate cortex (ACC). Single neurons within the ACC convey information about currently relevant SR mapping rules and signal unexpected action outcomes, which can be used to optimize behavioral choices. However, its functional significance and the mechanistic means of interaction with other nodes of the cognitive control network remain elusive and poorly understood. Here, we report that core aspects of cognitive control are encoded by rhythmic theta-band activity within neuronal circuits in the ACC. Throughout task performance, theta-activity predicted which of two SR mapping rules will be established before processing visual target information. Task-selective theta-activity emerged particularly early during those trials, which required the adjustment of SR rules following an erroneous rule representation in the preceding trial. These findings demonstrate a functional correlation of cognitive control processes and oscillatory theta-band activity in macaque ACC. Moreover, we report that spike output of a subset of cells in ACC is synchronized to predictive theta-activity, suggesting that the theta-cycle could serve as a temporal reference for coordinating local task selective computations across a larger network of frontal areas and the hippocampus to optimize and adjust the processing routes of sensory and motor circuits to achieve efficient sensory-motor control.

Unexpected action effects elicit deviance-related brain potentials and cause behavioral delay

To examine how people deal with perceivable consequences of their voluntary actions, we recorded event-related potentials (ERPs) during a self-paced, two-choice random generation task. Sixteen participants were asked to press one of two buttons randomly at a regular but self-selected interval of once per 1-2 s. Each button press produced either a 1000-Hz or 2000-Hz tone, but participants were told that the tones were irrelevant to the task. The button-tone combinations were initially fixed, but in subsequent blocks, a button press infrequently produced the tone associated with the opposite button (p=.15). This cognitively mismatched tone elicited N2, P3, and late positive potential (or positive slow wave) of the ERP and delayed the timing of the next button press. These results suggest that action effects are difficult to ignore and that an action effect that is different from a performer's expectation may cause task disruption.

Alteration of Radioprotective Effects of Heat-killed Lactobacillus casei in X-irradiated C3H/He Mouse Related to Blood Level of Proinflammatory Cytokines by Corticoids

It is well known that a pre-administration of proinflammatory cytokines alters hematopoietic progenitor cells to promote an increase resistance against radiation and increases the survival rate in mice irradiated with lethal doses of radiation. Inflammation stimulators, such as some bacterial constituents, are also reported to have similar radioprotective action. We found that pre-administration of heat-killed Lactobacillus casei (HLC) to mice increases the level of interleukin (IL)-1 beta in circulation as well as the survival rate following lethal dose of radiation. Since HLC stimulates early immune responses, effects by drugs to modify inflammation were studied. The increase of both blood IL-1 beta levels and survival rates by HLC were simultaneously accelerated by coadministration of mineralocorticoid and inhibited by glucocorticoids or corticotropin. Neither parameter was modified by non-steroidal anti-inflammatory or anti-rheumatoid drugs. This suggests that both expected radioprotective action and unexpected systemic action, realized as an increase in plasma cytokines, by inflammation-related radioprotectors can be controlled by the coadministration of drugs at least in C3H/He mice, based on consideration of their pharmacological properties.

Production of anArabidopsis hallerifoliar defensin inEscherichia coli

Production of the recombinant Arabidopsis halleri defensin AhPDF1.1 in a native-like form. Mature AhPDF1.1 cDNA was cloned into pET-28-a(+) and expressed in Escherichia coli Rosetta. After a denaturing extraction, purification by metal affinity chromatography and CNBr cleavage of the His-tag, a protein without extra amino acids at the N-terminus was obtained. An oxidative folding step was then required to renature the protein that was then purified to homogeneity by a C18 HPLC separation. Mass spectroscopy and circular dichroism analyses showed that the recombinant AhPDF1.1 has the expected molecular mass and 3D-structure features of a folded defensin with four-disulfide bridges. The recombinant protein is active against the filamentous fungus Fusarium oxysporum with a minimal inhibitory concentration of 0.6 micromol l(-1). The proposed purification protocol produces a native-like defensin suitable for tests of new biological roles. Plant defensins are essentially known as anti-fungal proteins; however, some unexpected actions on plant cells have recently been discovered. AhPDF1.1, for example, has been shown to confer zinc tolerance. Efficient production of native-like defensins is required to explore the different targets and roles of plant defensins.

Localizing GABA Receptors and Glutamate Transporters Using Conjugated Quantum Dots in Rat Cerebellum Slices

AbstractGABA and glutamate are known as the principal inhibitory and excitatory neurotransmitters in the vertebrate central nervous system, respectively. However, recent electro-physiological and immunogold data reported by Stell et al. [1] indicate that GABA may undergo also an excitatory action on presynaptic varicosities of parallel fibers (PFs) in the molecular layer of the rat cerebellum. PFs are axonal extensions, with a cross section of about 0.1 m, of the glutamatergic granule cells. Such an unexpected excitatory action of GABA indicates clearly the presence of GABA receptors in the PFs of granule cells. We show in this study that quantum dots may be used specifically and efficiently to label two endogenous synaptic proteins, namely R-GABAA-1 receptors (GABAA Rs) and glutamate transporters (VGLUT1) in order to target their localization in very small structures such as the presynaptic varicosities of the PFs, in agreement with the results recently reported by Stell et al..

Cangrelor Attenuates Coated-Platelet Formation

P2Y(12) inhibitors were introduced clinically as effective inhibitors of adenosine-5'-diphosphate (ADP) mediated platelet activation and aggregation. This class of pharmacological agents has enjoyed considerable success. Cangrelor is a recently developed P2Y(12) inhibitor that has the advantage of being an active drug not requiring metabolic conversion, although it is not orally available. Coated-platelets are a subclass of activated platelets generated on dual agonist activation with collagen plus thrombin; the primary hallmark of coated-platelets is their ability to support prothrombinase activity. Interestingly, we recently observed that the relatively weak agonist ADP potentiates the production of coated-platelets by the very strong agonists collagen plus thrombin, a previously unknown role for ADP. The authors sought in this study to determine if P2Y(12) inhibitors, such as cangrelor, were capable of attenuating this augmentation of coated-platelet generation. Cangrelor, at physiologically relevant concentrations, was able to eliminate the ADP-dependent increase in coated-platelet production with an IC(50) of 1.4 nM. Cangrelor, however, had no effect on thrombin-dependent platelet activation as measured by P-selectin expression. Although this in vitro study does not address the question of whether the effectiveness of cangrelor in vivo is partially due to an attenuation of coated-platelet production in addition to its documented antiaggregatory effects, it does reveal an unexpected action of cangrelor. Additional studies will be required to determine if all P2Y(12) inhibitors are equally effective in attenuating coated-platelet production.

Pleiotropic effects of selective CDK inhibitors on human normal and cancer cells

Escape from the proper control of the cell cycle by up-regulation of cyclins or aberrant activation of cyclin-dependent kinases (CDKs) as well as by inactivation of cellular inhibitors of CDKs (CKI) leads to malignant transformation. Loss of cellular CKIs in cancers provided a rationale for development of pharmacological inhibitors of CDKs. Recently synthesized CKIs, e.g., purine derivatives such as olomoucine (OLO) and roscovitine (ROSC) are non-genotoxic and exhibit increased selectivity towards CDK2 and CDK7/9. Interestingly, both drugs induce additional effects. Recently, a new, unexpected action of OLO on normal human cells was observed. OLO strongly up-regulates CLIMP-63, a 65kD protein that mediates the anchoring of the ER to microtubules. Moreover, ROSC induces in human MCF-7 cells phosphorylation of p53 protein at Ser-46 which in turn initiates caspase-independent apoptosis.In the present contribution we raised the question whether both CKIs would be able to block cell cycle progression and to reactivate p53 protein in human HPV-positive HeLa cervix carcinoma cells. We also addressed the question whether exponentially growing cancer cells are more susceptible to the inhibitory action of CKIs than normal cells. Our results show that HeLa cells are much more sensitive to ROSC than normal fibroblasts. ROSC induces G2 arrest and apoptosis in HeLa cells. It also reactivates and stabilizes wt p53 protein. The increase of p53 protein coincides with down-regulation of E6 oncoprotein.Thus, the biological action of substituted purines is not restricted to the inhibition of CDKs and open new perspectives for their therapeutic applications.

Answering the Call of the Court: How Justices and Litigants Set the Supreme Court Agenda. By Vanessa A. Baird. Charlottesville and London: Univ. of Virginia Press, 2007. Pp. xii+225. $45.00 cloth.

Answering the Call of the Court: How Justices and Litigants Set the Supreme Court Agenda. By Vanessa A. Baird. Charlottesville and London: Univ. of Virginia Press, 2007. Pp. xii+225. $45.00 cloth. In Answenng the Call of the Court: How Justices and Litigants Set the Supreme Court Agenda, Baird attempts to explain the relationship between justices and litigants early in the agenda-setting process. Baird implements a remarkable research design that yields a well-written volume. The primary theoretical contribution of this project is in the notion that in order to create social change, the Court depends on the support structure of extrajudicial players. Legal entrepreneurs pay heed to signals concerning justices' priorities and then sponsor cases in the appropriate issue areas. The cases sponsored present the type of legal questions and case facts that make them good vehicles for policy change. In other words, litigants have a pivotal role in translating shifts in the priorities of justices into actual changes in the agenda. The upshot of this symbiotic relationship is increased policymaking power to both justices and legal entrepreneurs. Agenda-setting, according to Baird, begins earlier than anybody else has ever argued. To a significant degree, it starts with politically salient decisions handed down by the Court approximately four years before the decision to grant certiorari in the current case. The pattern of the Court's changing interests is clear-when Baird employs different measures for justices' priorities, the result is that four to five years after the Court signals its interest in a certain issue area, the number of cases in this area on the Court's agenda swells significantly. Quantitative and qualitative analyses are sophisticatedly combined in this volume. Despite black-boxing some of the processes in her statistical analyses, Baird carefully outlines what happens in the box in the qualitative sections of the book. For instance, using various illustrative examples from Supreme Court cases, in Chapter 2 the author discusses the signals the Court sends to litigants. These include separate opinions registered by justices (e.g., the separate opinions in Furman v. Georgia [1972]) and unexpected actions (e.g., Justice Anthony Kennedy's decision to join the majority in Lawrence v. Texas [2003]). Baird also offers an in-depth analysis of how the Mexican American Legal Defense Fund (MALDEF) took advantage of legal cues in recent Supreme Court decisions to devise a legal strategy challenging the Personal Responsibility Act passed by Congress in 1996. In this case (Latino Coalition for a Healthy California v. Belshe [1997]), based on former Justice Sandra Day O'Connor's approach to federalism in New York v. United States (1992), the MALDEF chose federalism over equal protection as grounds for litigation. …

Unexpected actions of vitamin D: new perspectives on the regulation of innate and adaptive immunity

Knowledge about the ability of vitamin D to function outside its established role in skeletal homeostasis is not a new phenomenon. Nonclassical immunomodulatory and antiproliferative responses triggered by active 1,25-dihydroxyvitamin D were first reported more than a quarter of a century ago. It is only in recent years, however, that there has been a significant improvement in our understanding of how these nonclassical effects of vitamin D can influence the pathophysiology and possible prevention of human disease. Three particular strands of evidence have been prominent: firstly, population studies have revised our interpretation of normal vitamin D status in humans, suggesting, in turn, that vitamin D insufficiency is a clinical problem of global proportions; secondly, epidemiology has linked vitamin D status with disease susceptibility and/or mortality; and, thirdly, expression of the machinery required to synthesize 1,25-dihydroxyvitamin D in normal human tissue seems to be much more widespread than originally thought. Collectively, these observations suggest that nonclassical metabolism and response to vitamin D might have a significant role in human physiology beyond skeletal and calcium homeostasis. Specific examples of this will be detailed in the current Review, with particular emphasis on the immunomodulatory properties of vitamin D.

Exploiting Complexity and the Robustness of Network Architecture for Drug Discovery

The issue of complexity stands at the center of contemporary drug discovery and development. The central problem in drug development today is attrition of drug candidates identified by the modern molecular target-based discovery approach, due to two related features of complex metabolic networks: their fundamentally unpredictable response to targeted interventions and their "robustness" (tendency to maintain stable function in the face of internal or external perturbations). Complexity and adaptations are, therefore, generally seen as obstacles to drug discovery. Here, the converse proposition is presented-that the complexity and adaptive responses of highly interconnected metabolic networks can be exploited for therapeutic discovery. Unanticipated connectivity relationships may result in "off-target" changes in metabolic fluxes, leading to unexpected therapeutic actions of agents. Exploiting this approach requires that fully assembled living systems (in vivo models) be studied and that informative in vivo biomarkers of the activity of biochemical pathways responsible for disease be available. These biomarkers should be sensitive, predictive of functional endpoints, and have high enough throughput for efficient screening of large numbers of agents. To the extent that such biomarkers unambiguously reflect the activity of pathways that mediate disease or therapeutic response (i.e., are "authentic"), their utility will be increased. Examples are presented of pathway-based screening of approved drugs for unexpected actions. Results support the principle that agents that have one action typically have many actions, including unanticipated actions, reflecting connectivity relationships of complex networks. Pathway-based screening in vivo represents an alternative to the high attrition of the molecular target-based discovery paradigm.