Anthrax in transit; practical experience and intellectual exchange

Focusing on three Anglo-American outbreaks of industrial anthrax, this essay engages the question of how local circumstances influenced the transmission of scientific knowledge in the late nineteenth century. Walpole (Massachusetts), Glasgow, and Bradford (Yorkshire) served as important nodes of transnational investigation into anthrax. Knowledge about the morphology and behavior of Bacillus anthracis changed little while in transit between these nodes, even during complex debates about the nature of bacterial morphology, disease causation, and spontaneous generation. Working independently of their more famous counterparts (Robert Koch and Louis Pasteur), Anglo-American anthrax investigators used visual representations of anthrax bacilli to persuade their peers that a specific, identifiable cause produced all forms of anthrax-malignant pustule (cutaneous anthrax), intestinal anthrax, and woolsorter's disease (pneumonic anthrax). By the late 1870s, this point of view also supported what we would today call an ecological notion of the disease's origins in the interactions of people, animals, and microorganisms in the context of global commerce.     

Rare cell isolation and profiling on a hybrid magnetic/size-sorting chip

We present a hybrid magnetic/size-sorting (HMSS) chip for isolation and molecular analyses of circulating tumor cells (CTCs). The chip employs both negative and positive cell selection in order to provide high throughput, unbiased CTC enrichment. Specifically, the system utilizes a self-assembled magnet to generate high magnetic forces and a weir-style structure for cell sorting. The resulting device thus can perform multiple functions, including magnetic depletion, size-selective cell capture, and on-chip molecular staining. With such capacities, the HMSS device allowed one-step CTC isolation and single cell detection from whole blood, tested with spiked cancer cells. The system further facilitated the study of individual CTCs for heterogeneity in molecular marker expression.Circulating tumor cells (CTCs) have emerged as an important biomarker in clinical practice as well as in fundamental research.^{1, 2} CTCs, shed from primary tumors, have been shown to be an early harbinger of tumor expansion and metastasis³ and have been used to predict disease progression, response to treatment, relapse, and overall survival.^{4, 5, 6} Recent work has shown that CTCs display distinct proteomic and genetic profiles; for example, CTCs in pancreatic cancer, have increased RNA expression of Wnt, implicating this pathway in metastasis.⁷ Proteomic characterization of proliferative markers such as Ki-67, and hormonal markers such as androgen receptor in prostate cancer, also have been shown to be predictive of treatment outcome.^{8, 9}Despite such clinical potential of CTCs, their routine detection and characterization still remains a significant technical challenge.¹⁰ The task requires screening of a large number of cells (e.g., > 10⁷ cells in 10 ml blood) and enrichment of heterogeneous targets against a complex biological background. Two main methods of CTC isolation are typically used: positive and negative selection. In positive selection, CTCs are directly isolated from blood via size-based filtration^{11, 12, 13, 14, 15, 16, 17, 18, 19, 20} or antibody-based capture.^{1, 8, 21} Negative depletion reduces abundant blood cells, often by immunomagnetic separation, for downstream CTC enrichment.²² Both approaches have been used for high throughput CTC isolation from whole blood (SI Table 1).²³ Each method, however, has its own inherent limitations. Positive enrichment could be biased by its selection criteria (e.g., cell size and cell surface markers). Negative selection, albeit unbiased, often requires complex sample processing (e.g., multiple washing steps for CTC isolation) that could result in cell loss.We hypothesized that both positive and negative selection could be combined in a single platform to enable (1) highly efficient and unbiased CTC purification, and (2) in-situ molecular analyses of collected cells. As a proof-of-concept, we herein describe a hybrid magnetic/size-sorting (HMSS) system that integrates magnetic and size-based isolation into a compact microfluidic chip. The HMSS first uses a magnetic filter to deplete leukocytes through immunomagnetic capture. Samples then pass through a size-sorter region that traps individual cells at predefined locations. Since abundant leukocytes are removed by the magnetic filter, the size-sorter could have a low size cut-off (∼5 μm), which allows for the unbiased capture of even small cancer cells. Furthermore, molecular probes can be introduced to perform on-chip, multiplexed analyses at single-cell resolution. We evaluated the utility of the developed system by capturing and profiling tumor cells in whole blood. The HMSS offers the advantages of both negative and positive selection and thereby differs from the recently reported iChip system²⁴ which can operate only in either a negative or a positive selection mode.     

Resampling and Distribution of the Product Methods for Testing Indirect Effects in Complex Models

Recent advances in testing mediation have found that certain resampling methods and tests based on the mathematical distribution of 2 normal random variables substantially outperform the traditional z test. However, these studies have primarily focused only on models with a single mediator and 2 component paths. To address this limitation, a simulation was conducted to evaluate these alternative methods in a more complex path model with multiple mediators and indirect paths with 2 and 3 paths. Methods for testing contrasts of 2 effects were evaluated also. The simulation included 1 exogenous independent variable, 3 mediators and 2 outcomes and varied sample size, number of paths in the mediated effects, test used to evaluate effects, effect sizes for each path, and the value of the contrast. Confidence intervals were used to evaluate the power and Type I error rate of each method, and were examined for coverage and bias. The bias-corrected bootstrap had the least biased confidence intervals, greatest power to detect nonzero effects and contrasts, and the most accurate overall Type I error. All tests had less power to detect 3-path effects and more inaccurate Type I error compared to 2-path effects. Confidence intervals were biased for mediated effects, as found in previous studies. Results for contrasts did not vary greatly by test, although resampling approaches had somewhat greater power and might be preferable because of ease of use and flexibility.     

Making it happen: Possibilities and pitfalls of Cunningham's educational semiotic

In fundamental agreement with Cunningham's overall thesis, we attempt in this paper to articulate what an educational semiotic might look like on a practical level. For educational psychologists, it should mean an increased awareness of the limited, localized world views offered by current models of cognition and an increased attentiveness to alternate models. For teachers, an educational semiotic offers integration of cognition and affect and instructional strategies that depend upon and facilitate many ways of knowing. For an educational semiotic to be realized, however, there must be much greater attention paid to the roles of motivation and metacognition in semiotic pedagogy.     

Classically conditioned heart rate and respiratory-motor activity in newborn and neonatal pygmy goats

Classical discrimination conditioning of heart rate (HR) and respiratory-motor (RM) activity was examined in 16 newborn (average age = 1 day) and 8 neonatal (average age = 2 weeks) pygmy goats over 4 days of conditioning involving an electric shock US and auditory CSs. On the 4th day, one-half of the animals in each group received a low to moderate dose of ethanol to test the resistance of the CRs to drug effects. Discriminated conditioning of RM activity but not HR occurred in the newborn group, indicating an absence of close coupling between skeletal-motor activity and HR at this early age. While the newborn group failed to show recognizable HR reactions to the CSs during the 4 days of testing, it did show a long-latency brady-cardia UR beginning at 2 days of age that may have been secondary to baroreceptor reflex actions. The neonatal group displayed conditioning of both HR and RM activity. The similar latencies of the two types of CRs during the CS suggested that they may have been related to each other. However, the fact that the RM CR developed a day before the HR CR, combined with the observation that ethanol modified the HR CR but not the RM CR, supports the view that there was considerable independence between the two types of CRs.