Transcription factors E2F1 and E2F3 are expressed in placenta but do not regulate MMP14.

INTRODUCTION: Preeclampsia is a serious complication of pregnancy for which there are no efficacious medical treatments. Soluble endoglin is as an anti-angiogenic factor that contributes to the pathogenesis of the disease, however little is known about its molecular regulation in placenta. Recent data has demonstrated E2F transcription factors directly regulate MMPs in metastatic disease. Of particular interest was the capacity of E2F1 and E2F3 to up-regulate MMP14, a protease that cleaves and releases soluble endoglin from placenta. The aim of this study was to characterize E2F1 and E2F3 in preeclamptic placenta and assess whether silencing affects soluble endoglin release. METHODS: E2F1 and E2F3 mRNA, protein expression and localization were assessed in severe early onset preeclamptic and preterm control placentas (delivered <34 weeks gestation). E2F siRNA was administered to primary trophoblast and primary endothelial cells and effect on MMP14 mRNA expression and soluble endoglin secretion assessed. RESULTS: E2F1 and E2F3 were localized to the syncytiotrophoblast. E2F1 was significantly down regulated in severe preeclamptic placentas, while E2F3 was unchanged. Silencing E2F1 did not decrease MMP14 expression in primary trophoblast or endothelial cells. However, E2F1 silencing resulted in a significant increase in soluble endoglin secretion from both cell types, and silencing of E2F3 also significantly increased soluble endoglin release from primary trophoblast. DISCUSSION: This study demonstrates that E2F1 and E2F3 are present within the syncytiotrophoblast of placenta and that E2F1 is reduced in preeclampsia. Although silencing of either E2F1 or E2F3 does not alter MMP14 expression, both appear to regulate soluble endoglin release.

Placental SEMA3B expression is not altered in severe early onset preeclampsia.

Recent evidence suggests that Semaphorin 3B (SEMA3B) is upregulated in severe preeclampsia, and a major driver of cytotrophoblast aberrations in this disease. Here we independently assess whether SEMA3B expression is altered in a large cohort of severe early onset preeclamptic placentas. We demonstrate that SEMA3B relative mRNA expression and copy number are not changed in PE placentas. We confirm this at the protein level by western blot. Interestingly, exposure of term trophoblasts or explants to hypoxia induced a significant down regulation of SEMA3B mRNA, but a trend towards increased SEMA3B protein expression. We conclude that SEMA3B mRNA and protein is not altered in severe early onset preeclamptic placentas.

Stability of absolute copy number of housekeeping genes in preeclamptic and normal placentas, as measured by digital PCR.

Measuring mRNA expression is fundamental to placental research. Ideally, mRNA transcript numbers are directly quantified. However, PCR analysis using the ΔΔCT method relies on the stability of housekeeping genes and only reports relative expression. Digital PCR (dPCR) directly quantifies mRNA copy number and is more accurate than quantitative PCR. We quantified absolute mRNA copy number of housekeeping genes in normotensive pre-term (n = 20), severe preeclamptic (n = 11) and term (n = 12) placenta using dPCR. Whilst there was some variation, we confirm absolute mRNA copy number of GAPDH, TOP1, CYC1 and YWHAZ in placenta does not significantly alter between these cohorts, or across gestation.

The effect of pre-existing maternal obesity and diabetes on placental mitochondrial content and electron transport chain activity.

INTRODUCTION: Mitochondria dysfunction has been extensively implicated in the progression of these metabolic disorders, their role in placental tissue of diabetic and/or obese pregnant women is yet to be investigated. The aim of this study was to determine the effect of pre-existing type 1 and type 2 diabetes mellitus (DM), and pre-existing maternal obesity on placental mitochondrial function as assessed by mitochondrial content, electron transport chain (ETC) complex activities and oxidative stress. METHODS: Human placenta was obtained at the time of term Caesarean section from (i) non-obese (n = 19) and obese (n = 23) normal glucose tolerant (NGT) pregnant women; (ii) women with type 1 DM (n = 14) and BMI-matched NGT women (n = 14); and (iii) women with type 2 DM (n = 11) and BMI-matched NGT women (n = 11). The following endpoints were assessed: placental mitochondrial content by citrate synthase activity and mitochondrial DNA (mtDNA content); mitochondrial respiratory chain activity (complexes I, II, II & III, III and IV), and mitochondrial ROS (as assessed by mitochondrial hydrogen peroxide (H2O2) levels). RESULTS: When compared to placenta from NGT non-obese women, there was significantly lower mitochondrial DNA (mtDNA) content and electron transport chain complex I activity, and significantly higher mitochondrial H2O2 levels in placenta from NGT obese women (P < 0.05). Placental tissue from type 1 DM women showed significant reductions in ETC complex I, II & III, and III activity and increased H2O2 levels when compared to BMI-matched NGT women (P < 0.05). Type 2 DM women only exhibited significantly reduced ETC complex II & III activity when compared to BMI-matched NGT women (P < 0.05). DISCUSSION AND CONCLUSIONS: Women with pre-existing obesity or diabetes have decreased placental mitochondrial respiratory chain enzyme activities which may have detrimental consequences on placental function and therefore fetal growth and development.

Magnetic reconnection triggering magnetohydrodynamic instabilities during a sawtooth crash in a Tokamak plasma.

Thomson scattering measurements with subcentimeter spatial resolution have been made during a sawtooth crash in a Mega Ampere Spherical Tokamak fusion plasma. The unparalleled resolution of the temperature profile has shed new light on the mechanisms that underlie the sawtooth. As magnetic reconnection occurs, the temperature gradient at the island boundary increases. The increased local temperature gradient is sufficient to make the helical core unstable to ideal magnetohydrodynamic instabilities, thought to be responsible for the rapidity of the collapse.

Model for current-driven edge-localized modes.

Edge-localized modes (ELMs) are cyclic disturbances in the outer region of tokamak plasmas that are influential in determining present and future tokamak performance. In this Letter, we outline an approach to modeling ELMs in which we envisage toroidal peeling modes initiating a Taylor relaxation [Phys. Rev. Lett. 33, 1139 (1974)10.1103/PhysRevLett.33.1139] of a tokamak outer region plasma. Relaxation produces a peeling destabilizing flattened edge current profile and a stabilizing plasma-vacuum current sheet; the balance between the two determines the radial extent of the relaxed region. The model can be used to predict the energy losses due to an ELM and reproduces experimentally observed variations with edge safety factor and plasma collisionality. There is an intrinsic "deterministic scatter" in the model that also accords with observation.

Nonlinear saturation of tearing mode islands.

New, rigorous results for the tearing island saturation problem are presented. These results are valid for the realistic case where the magnetic island structure is non-symmetric about the reconnection surface and the electron temperature, on which the electrical resistivity depends, is evolved self-consistently with the island growth.

Stability of internal transport barriers to ideal MHD ballooning modes.

Internal transport barriers (ITB) in tokamaks can form near a minimum in the q profile, q(min), where magnetic shear is weak. We have analyzed their stability to short wavelength (n>>1, where n is the toroidal mode number) ideal MHD ballooning modes, by considering the s-alpha model equilibrium. We show that the ballooning transformation fails in regions of low shear but that one can then adopt a complementary approach based on the recurrence relation describing the toroidal coupling of radially localized modes on adjacent rational surfaces. Inclusion of the stabilizing effects of favorable average curvature or finite-n using this technique leads to stable high-pressure ITB configurations. The theory also shows the advantages of operating with low-order rational values of q(min).

Causal knowledge and categories: the effects of causal beliefs on categorization, induction, and similarity.

Despite the recent interest in the theoretical knowledge embedded in human representations of categories, little research has systematically manipulated the structure of such knowledge. Across four experiments this study assessed the effects of interattribute causal laws on a number of category-based judgments. The authors found that (a) any attribute occupying a central position in a network of causal relationships comes to dominate category membership, (b) combinations of attribute values are important to category membership to the extent they jointly confirm or violate the causal laws, and (c) the presence of causal knowledge affects the induction of new properties to the category. These effects were a result of the causal laws, rather than the empirical correlations produced by those laws. Implications for the doctrine of psychological essentialism, similarity-based models of categorization, and the representation of causal knowledge are discussed.

Problems for judgment and decision making.

This review examines recent developments during the past 5 years in the field of judgment and decision making, written in the form of a list of 16 research problems. Many of the problems involve natural extensions of traditional, originally rational, theories of decision making. Others are derived from descriptive algebraic modeling approaches or from recent developments in cognitive psychology and cognitive neuroscience.

The moderating influence of variability on belief revision.

The variability of the information on which a belief was initially based should moderate the degree to which the belief is revised when new disconfirming information is received. Reasoning from everyday intuitions, psychological theories of concept representation, and a rational mathematical analysis, we inferred that as variability increased, resistance to disconfirming information would also increase. Participants' judgments of the central tendencies of distributions of numerical information confirmed this prediction.

Reasoning in explanation-based decision making.

A general theory of explanation-based decision making is outlined and the multiple roles of inference processes in the theory are indicated. A typology of formal and informal inference forms, originally proposed by Collins (1978a, 1978b), is introduced as an appropriate framework to represent inferences that occur in the overarching explanation-based process. Results from the analysis of verbal reports of decision processes are presented to demonstrate the centrality and systematic character of reasoning in a representative legal decision-making task.

Tracing the footsteps of Sherlock Holmes: cognitive representations of hypothesis testing.

A well-documented phenomenon in opinion-revision literature is subjects' failure to revise probability estimates for an exhaustive set of mutually exclusive hypotheses in a complementary manner. However, prior research has not addressed the question of whether such behavior simply represents a misunderstanding of mathematical rules, or whether it is a consequence of a cognitive representation of hypotheses that is at odds with the Bayesian notion of a set relationship. Two alternatives to the Bayesian representation, a belief system (Shafer, 1976) and a system of independent hypotheses, were proposed, and three experiments were conducted to examine cognitive representations of hypothesis sets in the testing of multiple competing hypotheses. Subjects were given brief murder mysteries to solve and allowed to request various types of information about the suspects; after having received each new piece of information, subjects rated each suspect's probability of being the murderer. Presence and timing of suspect eliminations were varied in the first two experiments; the final experiment involved the varying of percentages of clues that referred to more than one suspect (for example, all of the female suspects). The noncomplementarity of opinion revisions remained a strong phenomenon in all conditions. Information-search data refuted the idea that subjects represented hypotheses as a Bayesian set; further study of the independent hypotheses theory and Shaferian belief functions as descriptive models is encouraged.

Ultrastructure of human basophil leukocytes studied after spray freezing and freeze-substitution.

Normal human blood cells were prepared for transmission electron microscopy by spray freezing, freeze-substitution and embedment in Araldite. Spray freezing was found to damage many cells. Nevertheless, the method developed preserved some leukocytes well and spray frozen, freeze-substituted human basophils showed interesting differences from conventionally prepared cells. Surface vesicles and tubulovesicular structures, extragranular myelin figures, and membranous projections on putative thin membrane-bounded granules were all absent. Preservation of the basophilic granules was clearly improved and their membranes were usually intact. The bulk content of these granules did not look particulate, but showed a very fine texture. Intragranular myelin figure material was well preserved and often enclosed striking lucent areas. Charcot-Leyden crystals were clearly seen in basophils from some donors, always in well preserved granules. A fourth type of material, not seen in conventionally prepared human basophils, was also often observed in the granules. Large lucent vacuoles could be regularly distinguished from basophilic granules. The granules with partly concave outlines, seen in conventionally prepared human basophils to be associated with ill-defined lucent areas and irregularity or projections of the membrane, were absent. In freeze-substituted cells, images suggesting fusion of basophilic granules with each other were seen quite frequently. It is concluded that, while the techniques needed to be improved, rapid freezing and freeze-substitution afford a better understanding of the ultrastructure of human basophil leukocytes.

Another look at sex stereotypes and social judgments: an analysis of the social perceiver's use of subjective probabilities.

Locksley, Borgida, Brekke, and Hepburn (1980) assert that subjects fall prey to the base-rate fallacy when they make stereotype-related trait judgments. They found that subjects ignored their stereotypes when trait judgments were made in the presence of trait-related behavioral information. The present article reexamines those findings with respect to two issues: (a) the use of a normative criterion in comparison with subjects' judgments and (b) the level of analysis (group vs. individual) of subjects' judgments. We conducted a replication of the Locksley et al. (1980) Study 2, and the results were examined with respect to these two issues. We found no support for the base-rate fallacy. When a Bayesian normative criterion was constructed for each subject based on the subject's own stereotype judgments and was compared with assertiveness judgments made in the presence of individuating information, there was no evidence that subjects ignored or underused their stereotypes as the base-rate fallacy predicts.

Individual differences in the use of simplification strategies in a complex decision-making task.

Decision time results were used to assess the strategies that 90 college undergraduates used in a complex decision-making task. Trend analyses revealed that the functions relating choice time to the number of choice alternatives in a set and the number of attributes comprising those alternatives contained linear (increasing) components. In addition, for a portion of the subjects, there was a quadratic effect of the number of attributes available to the decision maker on choice time, suggesting that these subjects adopted simplification strategies at high levels of task complexity. Reliable individual differences in these trend components were observed, consistent with individual differences in motivation and/or processing capacities. These individual differences were included in an information-processing model of decision behavior that captured the choice time data observed in this study. Subjects' ratings of apartments were used as a basis to assess the extent to which the use of simplification strategies resulted in preference reversals. Contrary to expectation, subjects whose choice times contained quadratic components demonstrated fewer preference reversals at high levels of information load.

Dissociation of the site of origin from the site of cryo-termination of ventricular tachycardia.

We performed conventional electrogram mapping and cryomapping in dogs with one-week-old experimentally-induced myocardial infarctions and programmed stimulation-induced sustained ventricular tachycardias to assess whether there is a correlation between the "site of origin" and site of cryo-termination of ventricular tachycardia (VT). Electrogram maps showed that 4 of 8 induced sustained VTs were due to macro- and 4 of 8 to microre-entry. Local cooling of the site of origin terminated 4 of 4 microre-entrant VTs, but only 1 of 4 macrore-entrant VTs. In the other 3 macrore-entrant VTs, the sites of cryo-termination were 2, 2.5, and 4 cm distant from the sites of origin. In contrast, cooling of the mid-to-late diastolic portions of the re-entry loops terminated all 8 VTs. These data demonstrate a dissociation of the site of origin from the site of cryo-termination of macrore-entrant VT.