In vitro to in vivo extrapolation from three-dimensional hiPSC-derived cardiac microtissues and physiologically based pharmacokinetic modeling to inform next-generation arrhythmia risk assessment.

Proarrhythmic cardiotoxicity remains a substantial barrier to drug development as well as a major global health challenge. In vitro human pluripotent stem cell-based new approach methodologies have been increasingly proposed and employed as alternatives to existing in vitro and in vivo models that do not accurately recapitulate human cardiac electrophysiology or cardiotoxicity risk. In this study, we expanded the capacity of our previously established three-dimensional human cardiac microtissue model to perform quantitative risk assessment by combining it with a physiologically based pharmacokinetic model, allowing a direct comparison of potentially harmful concentrations predicted in vitro to in vivo therapeutic levels. This approach enabled the measurement of concentration responses and margins of exposure for two physiologically relevant metrics of proarrhythmic risk (ie, action potential duration and triangulation assessed by optical mapping) across concentrations spanning three orders of magnitude. The combination of both metrics enabled accurate proarrhythmic risk assessment of four compounds with a range of known proarrhythmic risk profiles (ie, quinidine, cisapride, ranolazine, and verapamil) and demonstrated close agreement with their known clinical effects. Action potential triangulation was found to be a more sensitive metric for predicting proarrhythmic risk associated with the primary mechanism of concern for pharmaceutical-induced fatal ventricular arrhythmias, delayed cardiac repolarization due to inhibition of the rapid delayed rectifier potassium channel, or hERG channel. This study advances human induced pluripotent stem cell-based three-dimensional cardiac tissue models as new approach methodologies that enable in vitro proarrhythmic risk assessment with high precision of quantitative metrics for understanding clinically relevant cardiotoxicity.

Cardiotropic Isolates of Listeria monocytogenes with Enhanced Vertical Transmission Dependent upon the Bacterial Surface Protein InlB.

Listeria monocytogenes is a facultative Gram-positive intracellular bacterium that is capable of causing serious invasive infections in pregnant women, resulting in abortion, still-birth, and disseminated fetal infection. Previously, a clinical L. monocytogenes isolate, 07PF0776, was identified as having an enhanced ability to target cardiac tissue. This tissue tropism appeared to correlate with amino acid variations found within internalin B (InlB), a bacterial surface protein associated with host cell invasion. Given that the mammalian receptor bound by InlB, Met, is abundantly expressed by placental tissue, we assessed isolate 07PF0776 for its ability to be transmitted from mother to fetus. Pregnant Swiss Webster mice were infected on gestational day E13 via tail vein injection with the standard isolate 10403S, a noncardiotropic strain, or 07PF0776, the cardiac isolate. Pregnant mice infected with 07PF0776 exhibited significantly enhanced transmission of L. monocytogenes to placentas and fetuses compared to 10403S. Both bacterial burdens and the frequency of placental and fetal infection were increased in mice infected with the cardiac isolate. Strain 07PF0776 also exhibited an enhanced ability to invade Jar human trophoblast tissue culture cells in comparison to 10403S, and was found to have increased levels of InlB associated with the bacterial cell surface. Overexpression of surface InlB via genetic manipulation was sufficient to confer enhanced invasion of the placenta and fetus to both 10403S and 07PF0776. These data support a central role for surface InlB in promoting vertical transmission of L. monocytogenes.

Perils of the pneumatic tube: how clean are your pods?

Pneumatic tube systems (PTSs) are useful features in hospitals for efficient transport of items but further scrutiny reveals their potential risks. We investigated the extent of contamination of pods, used within the PTS with specific alert organisms, namely meticillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE) and carbapenemase-producing Enterobacterales (CPE). Results revealed contamination with VRE (53%) and MRSA (3%), which were reduced to only 3% (VRE) and 0% (MRSA) following disinfection. However recontamination occurred quickly following use. Our findings indicate that PTS could be an efficient method of transfer of potential pathogens around the hospital.

Assessing bacterial invasion of cardiac cells in culture and heart colonization in infected mice using Listeria monocytogenes.

Listeria monocytogenes is a Gram-positive facultative intracellular pathogen that is capable of causing serious invasive infections in immunocompromised patients, the elderly, and pregnant women. The most common manifestations of listeriosis in humans include meningitis, encephalitis, and fetal abortion. A significant but much less documented sequelae of invasive L. monocytogenes infection involves the heart. The death rate from cardiac illness can be up to 35% despite treatment, however very little is known regarding L. monocytogenes colonization of cardiac tissue and its resultant pathologies. In addition, it has recently become apparent that subpopulations of L. monocytogenes have an enhanced capacity to invade and grow within cardiac tissue. This protocol describes in detail in vitro and in vivo methods that can be used for assessing cardiotropism of L. monocytogenes isolates. Methods are presented for the infection of H9c2 rat cardiac myoblasts in tissue culture as well as for the determination of bacterial colonization of the hearts of infected mice. These methods are useful not only for identifying strains with the potential to colonize cardiac tissue in infected animals, but may also facilitate the identification of bacterial gene products that serve to enhance cardiac cell invasion and/or drive changes in heart pathology. These methods also provide for the direct comparison of cardiotropism between multiple L. monocytogenes strains.

PrfA-like transcription factor gene lmo0753 contributes to L-rhamnose utilization in Listeria monocytogenes strains associated with human food-borne infections.

Listeria monocytogenes is a food-borne bacterial pathogen and the causative agent of human and animal listeriosis. Among the three major genetic lineages of L. monocytogenes (i.e., LI, LII, and LIII), LI and LII are predominantly associated with food-borne listeriosis outbreaks, whereas LIII is rarely implicated in human infections. In a previous study, we identified a Crp/Fnr family transcription factor gene, lmo0753, that was highly specific to outbreak-associated LI and LII but absent from LIII. Lmo0753 shares two conserved functional domains, including a DNA binding domain, with the well-characterized master virulence regulator PrfA in L. monocytogenes. In this study, we constructed lmo0753 deletion and complementation mutants in two fully sequenced L. monocytogenes LII strains, 10403S and EGDe, and compared the flagellar motility, phospholipase C production, hemolysis, and intracellular growth of the mutants and their respective wild types. Our results suggested that lmo0753 plays a role in hemolytic activity in both EGDe and 10403S. More interestingly, we found that deletion of lmo0753 led to the loss of l-rhamnose utilization in EGDe, but not in 10403S. RNA-seq analysis of EGDe Δ0753 incubated in phenol red medium containing l-rhamnose as the sole carbon source revealed that 126 (4.5%) and 546 (19.5%) out of 2,798 genes in the EGDe genome were up- and downregulated more than 2-fold, respectively, compared to the wild-type strain. Genes related to biotin biosynthesis, general stress response, and rhamnose metabolism were shown to be differentially regulated. Findings from this study collectively suggested varied functional roles of lmo0753 in different LII L. monocytogenes strain backgrounds associated with human listeriosis outbreaks.

Genome sequence of Listeria monocytogenes 07PF0776, a cardiotropic serovar 4b strain.

Listeria monocytogenes is a food-borne bacterial pathogen commonly associated with serious invasive infections of the central nervous system or of the developing fetus. We present the genome sequence of Listeria monocytogenes 07PF0776, a serovar 4b isolate from a human myocardial abscess that exhibits enhanced invasion of cardiac tissue.

Second-order relational face processing is applied to faces of different race and photographic contrast.

A large body of research suggests that faces are processed by a specialized mechanism within the human visual system. This specialized mechanism is made up of subprocesses (Maurer, LeGrand, & Mondloch, 2002). One subprocess, called second- order relational processing, analyzes the metric distances between face parts. Importantly, it is well established that other-race faces and contrast-reversed faces are associated with impaired performance on numerous face processing tasks. Here, we investigated the specificity of second-order relational processing by testing how this process is applied to faces of different race and photographic contrast. Participants completed a feature displacement discrimination task, directly measuring the sensitivity to second-order relations between face parts. Across three experiments we show that, despite absolute differences in sensitivity in some conditions, inversion impaired performance in all conditions. The presence of robust inversion effects for all faces suggests that second-order relational processing can be applied to faces of different race and photographic contrast.

Actin polymerization drives septation of Listeria monocytogenes namA hydrolase mutants, demonstrating host correction of a bacterial defect.

The Gram-positive bacterial cell wall presents a structural barrier that requires modification for protein secretion and large-molecule transport as well as for bacterial growth and cell division. The Gram-positive bacterium Listeria monocytogenes adjusts cell wall architecture to promote its survival in diverse environments that include soil and the cytosol of mammalian cells. Here we provide evidence for the enzymatic flexibility of the murein hydrolase NamA and demonstrate that bacterial septation defects associated with a loss of NamA are functionally complemented by physical forces associated with actin polymerization within the host cell cytosol. L. monocytogenes ΔnamA mutants formed long bacterial chains during exponential growth in broth culture; however, normal septation could be restored if mutant cells were cocultured with wild-type L. monocytogenes bacteria or by the addition of exogenous NamA. Surprisingly, ΔnamA mutants were not significantly attenuated for virulence in mice despite the pronounced exponential growth septation defect. The physical force of L. monocytogenes-mediated actin polymerization within the cytosol was sufficient to sever ΔnamA mutant intracellular chains and thereby enable the process of bacterial cell-to-cell spread so critical for L. monocytogenes virulence. The inhibition of actin polymerization by cytochalasin D resulted in extended intracellular bacterial chains for which septation was restored following drug removal. Thus, despite the requirement for NamA for the normal septation of exponentially growing L. monocytogenes cells, the hydrolase is essentially dispensable once L. monocytogenes gains access to the host cell cytosol. This phenomenon represents a notable example of eukaryotic host cell complementation of a bacterial defect.

Norms for two types of manipulability (graspability and functional usage), familiarity, and age of acquisition for 320 photographs of objects.

There is increasing interest in the role that manipulability plays in processing objects. To date, Magnié, Besson, Poncet, and Dolisi's (2003) manipulability ratings, based on the degree to which objects can be uniquely pantomimed, have been the reference point for many studies. However, these ratings do not fully capture some relevant dimensions of manipulability, including whether an object is graspable and the extent to which functional motor associations above and beyond graspability are present. To address this, we collected ratings of these dimensions, in addition to ratings of familiarity and age of acquisition (AoA), for a set of 320 black-and-white photographs of objects. Familiarity and AoA ratings were highly correlated with previously reported ratings of the same dimensions (r = .853, p < .001, and r = .771, p < .001, respectively), validating the present norms. Grasping and functional use ratings, in contrast, were more moderately correlated with Magnié et al.'s pantomime manipulability ratings (r = .507, p < .001). These results were taken as evidence that the new manipulability ratings collected in this research capture distinct aspects of object manipulability. The complete stimuli and norms from this study may be downloaded from http://brm.psychonomic-journals.org/content/supplemental.

Changes to the object recognition system in patients with dementia of the Alzheimer's type.

Do DAT patients show category-specific deficits in object identification, and do they arise from semantic or visual damage? Participants decided whether line drawings of living and nonliving objects matched names at superordinate, basic, or subordinate levels. Patients were most impaired with superordinate decisions. Controls had most difficulty with subordinate decisions. No category-specific deficit was found with patients. Impaired superordinate decisions by the patients support semantic damage. If category-specific deficits arise from damaged semantics, they should have been found. Since they were not, and since patients performed subordinate decisions the best, a visual basis to category specificity is supported. Finally, a living advantage was found with normal observers which cannot be spurious due to differences in concept familiarity since living and nonliving objects were matched for this variable.

Prenatal care needs, availability, accessibility, use, and satisfaction: a comparison of military women within and outside the continental United States.

This article presents the results of a survey distributed to military women within and outside the continental United States (CONUS and OCONUS) who had received prenatal care and delivered at one of six Army, Navy, or Air Force military facilities. This comparative descriptive study was undertaken to determine from the mothers' perspective (1) their needs, availability, accessibility, use, satisfaction, and preferences for prenatal care services, and (2) if prenatal care and birth outcomes were significantly different for CONUS versus OCONUS women. Both groups reported a high degree of satisfaction with services, clinics/offices, and health care staff. Significant group differences were reported in transportation, length of travel time, and cost of traveling, with the CONUS group reporting greater problems with each. For both groups, quality and consistency of health care were the two most important factors in their preference of provider. More than 20% of the mothers reported receiving no information on some of the common concerns of pregnancy. There were no significant group differences with regard to infant birth weight or length of hospital stay.

Testing a two-component model of face identification: effects of inversion, contrast reversal, and direction of lighting.

Enns and Shore (1997 Perception & Psychophysics 59 23-31) found additive effects of test orientation (upright or inverted) and direction of lighting (brow or chin lit) when they studied the inversion effect on face identification. A two-stage model was inferred in which inversion was processed by an orientation-sensitive component after which chin-lighting was processed by a lighting-sensitive component. Face identification is also strongly influenced by contrast reversal. A study is reported which aimed to (i) determine if contrast reversal interacts with lighting direction or orientation, findings that would support Enns and Shore's model; and (ii) to test their assumption that holistic encoding is prerequisite for their model by inducing featural encoding through training names to inverted faces. Names for unfamiliar brow-lit positive-contrast faces were trained with the faces upright or inverted. Identification accuracy was measured with combinations of orientation, lighting, and contrast. Consistent with their model, test orientation and direction of lighting were additive after training on upright faces and lighting and contrast reversal interacted. When holistic encoding was prevented following training on inverted faces, test orientation and lighting direction interacted for positive-contrast faces. Negative faces showed only an effect of direction of lighting. These results support Enns and Shore's two-stage model and their interpretation that orientation and direction of lighting interact following featural encoding of faces.

Clinical challenges in menopausal management: a case study approach.

Selecting an appropriate hormone replacement regimen can pose many challenges to the clinician and the patient. This report offers 4 case studies and clinical considerations illustrating both scientific and clinical considerations when collaborating with women to find an acceptable method to manage menopausal symptoms. Both traditional and complementary therapies are considered.

Exploration of facilitators and barriers to prenatal care among military women.

Early and regular prenatal care has been shown to decrease the incidence of adverse pregnancy outcomes; however, little research has examined it experientially from the mother's perspective. The purpose of this qualitative study was to explore the experience of prenatal care from the military mother's perspective. Using purposive sampling, 16 women who had recently given birth were interviewed. The data were analyzed using a qualitative descriptive research methodology to elicit categories of substantive concepts, ideas, or themes. Data explored in this study included availability, use and satisfaction with prenatal care services, and barriers, preferences, and recommendations.

A guide to understanding informed consent.

Anesthesia providers are expected to provide information to the patient during the preanesthesia interview that enables the patient to make informed choices. Adequate disclosure during the informed consent process ensures the equalization of the practitioner/patient relationship and the decision-making rights of the patient. Both certified registered nurse anesthetists (CRNAs) and anesthesiologists are not only legally required to provide information that will allow a patient to make an informed judgment about how to proceed with various anesthetic modalities but are also obligated by their standards of practice. This article informs the CRNA about the principles of informed consent so that they can better understand their role in the informed consent process.

Telephone triage in women's health care.

This article describes the importance of sound telephone triage strategies in women's health care. A brief description of the history of telephone triage is provided as well as basic considerations in establishing a telephone triage system. A sample telephone report sheet is also provided.

Effects of orientation on the identification of rotated objects depend on the level of identity.

Matching names and rotated line drawings of objects showed effects of object orientation that depended on name level. Large effects, in the same range as object naming, were found for rotations between 0 degrees and 120 degrees from upright with subordinate names (e.g., collie), whereas nonsignificant effects were found with superordinate (e.g., animal) and basic names (e.g., dog). These results support image normalization, after contact with orientation-invariant representations, that provide basic-level identity. They consequently fail to support theories of object recognition in which rotated object images are normalized to the upright position before contact with long-term object representations.

Rotated object identification with and without orientation cues.

The time to name two-dimensional line drawings of objects increases linearly for object rotations between 0 degrees and 120 degrees from the upright. Several theories attribute these effects of orientation to finding the top or the top-bottom axis of objects. By this account, prior knowledge of the location of the top or the top-bottom axis of objects should diminish effects of object orientation when they are named. When this hypothesis was tested by cuing the top or the top-bottom axis, no reduction in the effects of orientation on object naming was found. This result is inconsistent with effects of orientation on object naming being due to finding the top or the top-bottom axis. Instead, the top may be found prior to rotational normalization of the object image.

Informed consent: an essential element of safe anesthesia practice.

Generally, health care providers have viewed safety in terms of prevention of patient accidents. However, with the growth of patient consumerism and stress on quality improvement, the concept of "safety" has been expanded. This article examines the legal concept of informed consent and offers practical suggestions on increasing both patient and provider safety and improving quality of care. For reasons dictated by statute, case law, and professional ethics, informed consent should be part of the practice of every CRNA. With proper informed consent, misinformation, dissatisfaction, and subsequent legal action can be diminished. Information should be offered to the patient and family and reinforced with written educational materials and instructions. These procedures should be documented in the medical record to provide verification that the patient was informed of the risks and benefits and agreed to the procedure contemplated. Failure to do so could expose the CRNA to legal actions under legal theories that include not only negligence, but battery, and contract as well.