Assessment of the Impact of Alternative Fixatives on HER2 Detection in Breast Cancer and Gastric Cancer Tumor Specimens.

The type of fixative used for preserving tumor specimens can significantly impact the performance of the immunohistochemistry and in situ hybridization assays used for assessing human epidermal growth factor receptor 2 (HER2) status. This study reports the prevalence of the use of alternative fixatives other than the guideline-recommended 10% neutral buffered formalin (NBF) during HER2 testing in a real-world setting. The effects of alternative fixatives [20% NBF and 10% unbuffered formalin (UBF) fixatives] on HER2 testing of breast cancer (BC) and gastric cancer (GC) cell lines and tissues are also assessed. Overall, 117,636 tumor samples received at a central laboratory from >8000 clinical trial sites across 60 countries were reviewed to determine the prevalence of alternative fixative usage. To investigate the impact of alternative fixatives, 27 cell lines (21 BC and 6 GC) and 76 tumor tissue samples (50 BC and 26 GC) were fixed in 10% NBF, 20% NBF, or 10% UBF, and evaluated for HER2 status by immunohistochemistry and in situ hybridization. Real-world data showed that 9195 (7.8%) tumor samples were preserved using an alternative fixative. In cell lines, overall percentage agreement, negative percentage agreement, and positive percentage agreement among the 3 fixatives were 100%. In tumor tissues, the agreement among 10% NBF, 20% NBF, and 10% UBF ranged between 94.7% and 96.6% for negative percentage agreement and 90.9% for overall percentage agreement compared with a range of 58.3% to 66.7% for positive percentage agreement. These results suggest that alternative fixatives may have the potential to convert HER2 status in tissues from positive to negative.

Do Environmental Cues to Discovery Influence the Likelihood to Rape?

Research on men's sexual exploitation of women has documented that men's psychology tracks cues associated with the ease of women's exploitability. In the current studies, we examined a different class of cues hypothesized to aid men's use of sexually exploitative strategies: environmental cues to the likelihood of discovery. We defined likelihood of discovery as the perceived probability of identification when engaging in exploitative behavior (e.g., presence of others). We test the hypothesis that men's likelihood to rape increases when their perception of the likelihood of discovery is low in three studies. In Study 1, we conducted a content analysis of individuals' responses (N = 1,881) when asked what one would do if they could stop time or be invisible. Besides the "other" category whereby there were no specific category for nominated behaviors, the most nominated category included sexually exploitative behavior-representing 15.3% of reported behaviors. Both Studies 2 (N = 672) and 3 (N = 614) were preregistered manipulations of likelihood of discovery surreptitiously testing men's rape likelihood to rape across varying levels of discovery. We found men, compared to women, reported a statistically higher likelihood to rape in both Studies 2 and 3: 48% compared to 39.7% and 19% compared to 6.8%, respectively. Across Studies 2 and 3, we found no statistical effect of the likelihood of discovery on participants' likelihood to rape. We discuss how the presence of one's peers may provide social protection against the costs of using an exploitative sexual strategy if a perpetrator is caught.

Is the Vertical-Horizontal Illusion a Byproduct of the Environmental Vertical Illusion?

The vertical-horizontal illusion is the overestimation of a vertical line compared to a horizontal line of the same length. Jackson and Cormack (2007) proposed that the vertical-horizontal illusion might be a byproduct of the mechanisms that generate the environmental vertical illusion, which is the tendency to overestimate vertical distances (i.e., heights) relative to horizontal distances the same length. In our study, 326 undergraduate participants stood atop an 18.6-meter parking structure and estimated both the height of the structure and the horizontal distance of a target placed 18.6 meters away, using a moveable horizontal target across the length of the structure. Participants also completed a vertical-horizontal illusion task by drawing a horizontal line below a 9.1 cm vertical line. We correlated vertical distance estimates with vertical line estimates to test Jackson and Cormack's byproduct hypothesis. This hypothesis was very weakly-if at all-supported by the data: Participants' overestimations in the vertical-horizontal illusion task explained 1% of the variance associated with their overestimations in the environmental vertical illusion task. Additionally, to test whether the environmental vertical illusion is impervious to explicit awareness, a random half of our participants were advised to be mindful that people tend to overestimate heights. The results supported our second hypothesis: Even when participants were made aware of the environmental vertical illusion, they still reliably overestimated heights. Discussion addressed implications for the robustness of the environmental vertical illusion (e.g., treatment of those with acrophobia).

A Phenotypic High-Throughput Screen with RSV-Infected Primary Human Small Airway Epithelial Cells (SAECs).

Respiratory syncytial virus (RSV) is a commonly occurring pathogen that can cause severe disease in children, the elderly, and immunocompromised individuals with a large, unmet clinical need. We developed a high-throughput, primary cell-based antiviral RSV assay to enable identification of small molecules using cytopathic effect (CPE) as a phenotypic end point. To provide increased biological relevance, we developed our assay with primary human small airway epithelial cells (SAECs), which originate from known sites of RSV infection and replication instead of a more traditional immortalized cell line. Using purchased low-passage cells, cost-effective large-scale culture methods were developed to provide assay-ready frozen SAECs. A high-throughput screening campaign using the GSK Screening Collection was performed. The screen was executed in 384-well plates over a 12-week period with an average Z' of 0.5. The screen yielded 17 post-entry hits with activity in the primary cells, which were not active in immortalized cells. Potencies for this class of compounds were equal between the primary and immortalize cell lines. For entry inhibitors, the number was much lower, with increased potency observed in immortalized cells. This is the first known use of frozen primary human cells for an RSV high-throughput screening phenotypic campaign.

The implicit rules of combat.

Conspecific violence has been pervasive throughout evolutionary history. The current research tested the hypotheses that individuals implicitly categorize combative contexts (i.e., play fighting, status contests, warfare, and anti-exploitative violence) and use the associated contextual information to guide expectations of combative tactics. Using U.S. and non-U.S. samples, Study 1 demonstrated consistent classification of combative contexts from scenarios for which little information was given and predictable shifts in the acceptability of combative tactics across contexts. Whereas severe tactics (e.g., eye-gouging) were acceptable in warfare and anti-exploitative violence, they were unacceptable in status contests and play fights. These results suggest the existence of implicit rules governing the contexts of combat. In Study 2, we explored the reputational consequences of violating these implicit rules. Results suggest that rule violators (e.g., those who use severe tactics in a status contest) are given less respect. These are the first studies to implicate specialized mechanisms for aggression that use contextual cues of violence to guide expectations and behavior.

Changes in sleep time and sleep quality across the ovulatory cycle as a function of fertility and partner attractiveness.

Research suggests that near ovulation women tend to consume fewer calories and engage in more physical activity; they are judged to be more attractive, express greater preferences for masculine and symmetrical men, and experience increases in sexual desire for men other than their primary partners. Some of these cycle phase shifts are moderated by partner attractiveness and interpreted as strategic responses to women's current reproductive context. The present study investigated changes in sleep across the ovulatory cycle, based on the hypothesis that changes in sleep may reflect ancestral strategic shifts of time and energy toward reproductive activities. Participants completed a 32-day daily diary in which they recorded their sleep time and quality for each day, yielding over 1,000 observations of sleep time and quality. Results indicated that, when the probability of conception was high, women partnered with less attractive men slept more, while women with more attractive partners slept less.

Oral sex, semen displacement, and sexual arousal: testing the ejaculate adjustment hypothesis.

Male Indian Flying Foxes (Pteropus giganteus) that spend more time performing oral sex on a female also spend more time copulating with her. In humans, men who spend more time copulating with their regular partner also perform more "semen-displacing" copulatory behaviors (e.g., deeper, more vigorous penile thrusting). We investigated whether men who spend more time performing oral sex on their regular partner also spend more time copulating with her and perform more semen-displacing copulatory behaviors. We proposed and tested the ejaculate adjustment hypothesis for men's copulatory behaviors: Men adjust their copulatory behaviors to increase their sexual arousal and consequent ejaculate quality, thereby increasing their chances of success in sperm competition. Two hundred and thirty-three men in a committed, heterosexual relationship responded to questions about their copulatory behavior and sexual arousal during their most recent sexual encounter with their long-term partner. The results indicated that men who spend more time performing oral sex on their partner also spend more time copulating with her, perform more semen-displacing copulatory behaviors, and report greater sexual arousal. We discuss limitations to the current research and highlight the heuristic value of sperm competition theory for understanding human sexual behaviors.

Characterization of a novel PERK kinase inhibitor with antitumor and antiangiogenic activity.

The unfolded protein response (UPR) is a signal transduction pathway that coordinates cellular adaptation to microenvironmental stresses that include hypoxia, nutrient deprivation, and change in redox status. These stress stimuli are common in many tumors and thus targeting components of the UPR signaling is an attractive therapeutic approach. We have identified a first-in-class, small molecule inhibitor of the eukaryotic initiation factor 2-alpha kinase 3 (EIF2AK3) or PERK, one of the three mediators of UPR signaling. GSK2656157 is an ATP-competitive inhibitor of PERK enzyme activity with an IC(50) of 0.9 nmol/L. It is highly selective for PERK with IC(50) values >100 nmol/L against a panel of 300 kinases. GSK2656157 inhibits PERK activity in cells with an IC(50) in the range of 10-30 nmol/L as shown by inhibition of stress-induced PERK autophosphorylation, eIF2α substrate phosphorylation, together with corresponding decreases in ATF4 and CAAT/enhancer binding protein homologous protein (CHOP) in multiple cell lines. Oral administration of GSK2656157 to mice shows a dose- and time-dependent pharmacodynamic response in pancreas as measured by PERK autophosphorylation. Twice daily dosing of GSK2656157 results in dose-dependent inhibition of multiple human tumor xenografts growth in mice. Altered amino acid metabolism, decreased blood vessel density, and vascular perfusion are potential mechanisms for the observed antitumor effect. However, despite its antitumor activity, given the on-target pharmacologic effects of PERK inhibition on pancreatic function, development of any PERK inhibitor in human subjects would need to be cautiously pursued in cancer patients.

Discovery of GSK2656157: An Optimized PERK Inhibitor Selected for Preclinical Development.

We recently reported the discovery of GSK2606414 (1), a selective first in class inhibitor of protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK), which inhibited PERK activation in cells and demonstrated tumor growth inhibition in a human tumor xenograft in mice. In continuation of our drug discovery program, we applied a strategy to decrease inhibitor lipophilicity as a means to improve physical properties and pharmacokinetics. This report describes our medicinal chemistry optimization culminating in the discovery of the PERK inhibitor GSK2656157 (6), which was selected for advancement to preclinical development.

Discovery of 7-methyl-5-(1-{[3-(trifluoromethyl)phenyl]acetyl}-2,3-dihydro-1H-indol-5-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine (GSK2606414), a potent and selective first-in-class inhibitor of protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK).

Protein kinase R (PKR)-like endoplasmic reticulum kinase (PERK) is activated in response to a variety of endoplasmic reticulum stresses implicated in numerous disease states. Evidence that PERK is implicated in tumorigenesis and cancer cell survival stimulated our search for small molecule inhibitors. Through screening and lead optimization using the human PERK crystal structure, we discovered compound 38 (GSK2606414), an orally available, potent, and selective PERK inhibitor. Compound 38 inhibits PERK activation in cells and inhibits the growth of a human tumor xenograft in mice.

Regulation of ingestive behaviors in the rat by GSK1521498, a novel micro-opioid receptor-selective inverse agonist.

µ-Opioid receptor (MOR) agonism induces palatable food consumption principally through modulation of the rewarding properties of food. N-{[3,5-difluoro-3'-(1H-1,2,4-triazol-3-yl)-4-biphenylyl]methyl}-2,3-dihydro-1H-inden-2-amine (GSK1521498) is a novel opioid receptor inverse agonist that, on the basis of in vitro affinity assays, is greater than 10- or 50-fold selective for human or rat MOR, respectively, compared with κ-opioid receptors (KOR) and δ-opioid receptors (DOR). Likewise, preferential MOR occupancy versus KOR and DOR was observed by autoradiography in brain slices from Long Evans rats dosed orally with the drug. GSK1521498 suppressed nocturnal food consumption of standard or palatable chow in lean and diet-induced obese (DIO) Long Evans rats. Both the dose-response relationship and time course of efficacy in lean rats fed palatable chow correlated with µ receptor occupancy and the plasma concentration profile of the drug. Chronic oral administration of GSK1521498 induced body weight loss in DIO rats, which comprised fat mass reduction. The reduction in body weight was equivalent to the cumulative reduction in food consumption; thus, the effect of GSK1521498 on body weight is related to inhibition of food consumption. GSK1521498 suppressed the preference for sucrose-containing solutions in lean rats. In operant response models also using lean rats, GSK1521498 reduced the reinforcement efficacy of palatable food reward and enhanced satiety. In conclusion, GSK1521498 is a potent, MOR-selective inverse agonist that modulates the hedonic aspects of ingestion and, therefore, could represent a pharmacological treatment for obesity and binge-eating disorders.

Perceived risk of female infidelity moderates the relationship between objective risk of female infidelity and sexual coercion in humans (Homo sapiens).

Female extrapair copulation (EPC) can be costly to a woman's long-term romantic partner. If a woman has copulated recently with a man other than her long-term partner, her reproductive tract may contain the sperm of both men, initiating sperm competition (whereby sperm from multiple males compete to fertilize an egg). Should the woman become pregnant, her long-term partner is at risk of cuckoldry-investing unwittingly in offspring to whom he is not genetically related. Previous research in humans (Homo sapiens) and in nonhuman animals suggests that males have evolved tactics such as partner-directed sexual coercion that reduce the risk of cuckoldry. The current research provides preliminary evidence that mated men (n = 223) at greater risk of partner EPC, measured as having spent a greater proportion of time apart from their partner since the couple's last in-pair copulation, more frequently perform partner-directed sexually coercive behaviors. This relationship is moderated, however, by men's perceived risk of partner EPC, such that the correlation between the proportion of time spent apart since last in-pair copulation and sexually coercive behaviors remains significant only for those men who perceive themselves to be at some risk of partner EPC. Discussion addresses limitations of this research and highlights directions for future research investigating the relationship between female EPC and men's partner-directed sexual coercion.

Tetrahydroquinoline derivatives as opioid receptor antagonists.

Opioid receptors play an important role in both behavioral and homeostatic functions. We herein report tetrahydroquinoline derivatives as opioid receptor antagonists. SAR studies led to the identification of the potent antagonist 2v, endowed with 1.58nM (K(i)) functional activity against the µ opioid receptor. DMPK data suggest that novel tetrahydroquinoline analogs may be advantageous in peripheral applications.

The evolutionary psychology of violence.

This paper reviews theory and research on the evolutionary psychology of violence. First, I examine evidence suggesting that humans have experienced an evolutionary history of violence. Next, I discuss violence as a context-sensitive strategy that might have provided benefits to our ancestors under certain circumstances. I then focus on the two most common forms of violence that plague humans -violence over status contests and intimate partner violence- outlining psychological mechanisms involved in each. Finally, I suggest that greater progress will be made by shifting the study from contexts to mechanisms.

Thienopyrimidine-based dual EGFR/ErbB-2 inhibitors.

Two new series of potent and selective dual EGFR/ErbB-2 kinase inhibitors derived from novel thienopyrimidine cores have been identified. Isomeric thienopyrimidine cores were evaluated as isosteres for a 4-anilinoquinazoline core and several analogs containing the thieno[3,2-d]pyrimidine core showed anti-proliferative activity with IC(50) values less than 1 microM against human tumor cells in vitro.

Sexual coercion in intimate relationships: a comparative analysis of the effects of women's infidelity and men's dominance and control.

Researchers studying the proximate (or immediate) causes of sexual coercion have proposed that partner rape is motivated by a man's attempt to dominate and control his partner and that this expression of power is the product of men's social roles. Researchers studying the ultimate (or evolutionary) causes, in contrast, have proposed that partner rape may function as an anti-cuckoldry tactic, with its occurrence related to a man's suspicions of his partner's sexual infidelity. In two studies, we collected data relevant to both perspectives to explore how these variables interact with men's sexual coercion in an intimate relationship. Regression analyses from Study 1 (self-reports from 256 men) and Study 2 (partner-reports from 290 women) indicated that men's sexual coercion of their partners was consistently predicted by female infidelity and men's controlling behavior, suggesting that both variables are necessary to explain men's sexual coercion. Discussion addressed limitations of the current research and highlighted the importance of integrating multiple levels of analysis when studying men's sexual coercion of their intimate partners.

Discovery of small molecule agonists for the bombesin receptor subtype 3 (BRS-3) based on an omeprazole lead.

Starting from a weak omeprazole screening hit, replacement of the pyridine with a 1,3-benzodioxole moiety, modification of the thioether linkage, and substitution of the benzimidazole pharmacophore led to the discovery of nanomolar BRS-3 agonists.

Absence makes the adaptations grow fonder: proportion of time apart from partner, male sexual psychology, and sperm competition in humans (Homo sapiens).

Sperm competition occurs when the sperm of multiple males concurrently occupy the reproductive tract of a female and compete to fertilize an egg. We used a questionnaire to investigate psychological responses to the risk of sperm competition for 237 men in committed, sexual relationships. As predicted, a man who spends a greater (relative to a man who spends a lesser) proportion of time apart from his partner since the couple's last copulation reported (a) greater sexual interest in his partner, (b) greater distress in response to his partner's sexual rejection, and (c) greater sexual persistence in response to his partner's sexual rejection. All effects were independent of total time since the couple's last copulation and the man's relationship satisfaction. Discussion addresses limitations of the current research and situates the current results within the broader comparative literature on adaptation to sperm competition.

Solid phase synthesis and SAR of small molecule agonists for the GPR40 receptor.

The discovery, synthesis and structure-activity relationship (SAR) of novel carboxylic acid agonists for GPR40 are described. Aryl propionic acid 1, identified from a high throughput screen, was selected for chemical exploration. Compound 2 was identified as our lead molecule through efficient solid phase combinatorial array chemistry and had an attractive in vitro and in vivo pharmacokinetic profile in rat. These ligands may prove useful in establishing a role for GPR40 in insulin regulation.

Potent, selective, and orally efficacious antagonists of melanin-concentrating hormone receptor 1.

The high expression of MCH in the hypothalamus with the lean hypophagic phenotype coupled with increased resting metabolic rate and resistance to high fat diet-induced obesity of MCH KO mice has spurred considerable efforts to develop small molecule MCHR1 antagonists. Starting from a lead thienopyrimidinone series, structure-activity studies at the 3- and 6-positions of the thienopyrimidinone core afforded potent and selective MCHR1 antagonists with representative examples having suitable pharmacokinetic properties. Based on structure-activity relationships, a structural model for MCHR1 was constructed to explain the binding mode of these antagonists. In general, a good correlation was observed between pKas and activity in the right-hand side of the template, with Asp123 playing an important role in the enhancement of binding affinity. A representative example when evaluated chronically in diet-induced obese mice resulted in good weight loss effects. These antagonists provide a viable lead series in the discovery of new therapies for the treatment of obesity.