Residency training and use of strategies to promote community child health one year after graduation.
Training pediatricians to understand child health in the context of community and to develop skills to engage with community organizations remain priorities for residency education in the United States. Our objectives were to: 1) determine use of strategies to influence community child health by graduates of the Community Pediatrics Training Initiative (CPTI); and 2) to identify personal, practice, and residency program factors associated with use of strategies 1 year after residency. Analysis of data from the Dyson Initiative National Evaluation included surveys of physicians ("graduates") 1 year after residency and surveys of CPTI program leaders. Graduates reported personal and practice characteristics and use of one or more strategies to influence community child health. Chi-square and logistic regression were used to examine associations between personal, practice, and programmatic factors with use of strategies. Of the 511 graduates (68% participation), 44% reported use of one or more strategies. After adjusting for residency site, time spent in general pediatrics, and program emphasis on individual level advocacy, we found that graduates were more likely to report using strategies if they felt responsible for improving community child health (adjusted odds ratio [aOR] 4.1, 95% confidence interval [95% CI] 2.5-6.9), had contact with a person who provides guidance about community pediatrics (aOR 1.8, CI 1.2-2.6), or trained in a program that places great emphasis on teaching population level advocacy skills (aOR 2.3, CI 1.3-4.2). Personal perspectives and residency education influence community involvement, even early in pediatricians' careers. Efforts are needed to understand how content and delivery of training influence community engagement over time.


[Morphological peculiarities of human anal glands during postnatal ontogenesis].
Macro-microscopical and histological methods were used to study the age-related peculiarities of human anal glands in 267 practically healthy subjects of both sexes of various ages (from newborns to 102 years). Topography of anal glands was described, their numbers and dimensional parameters were determined in the subjects of different age. Maximal total number of these glands, maximal length and width of their initial region, as well as maximal number of initial portions within it, were found at the age of 22-35 years. Subsequently, as the age increased, these parameters tended to decrease. Connective tissue fiber content in the glands was found to increase during postnatal ontogenesis. Along the course of common excurrent duct in submucosa and in mucosa, some dilated regions were found, especially in aged and senile persons.


Anaesthesia and chest wall function.
The effects of anaesthesia and surgery on the chest wall may be responsible for impaired gas exchange and other pulmonary complications during the perioperative period. Current evidence supports the following sequence of events. Anaesthesia changes the shape and motion of the chest wall, either by changing the amount of tonic and phasic activity of the respiratory muscles (anaesthesia with spontaneous breathing) or by eliminating the activity entirely (paralysis with mechanical ventilation). These primary changes in chest wall function lead to secondary changes in lung function, including the formation of dependent lung atelectasis. Recent advances in imaging of the thorax have called into question traditional understanding of the mechanisms by which anaesthesia alters chest wall function. For example, it is now apparent that anaesthesia reduces the functional residual capacity not by changing the position of the diaphragm, but rather by affecting the rib cage, and, perhaps, the volume of intrathoracic blood. The effects of anaesthesia and surgery on postoperative chest wall function may be lessened by regional analgesia and the use of laparoscopic surgical techniques. However, it is not yet clear that this improvement is associated with a reduction in the incidence of pulmonary complications.


Complications of lumbar spinal fluid drainage.
Cerebrospinal fluid fistula is an unfortunate, yet well-recognized, complication of basilar skull fracture, skull base surgery, and neurotologic procedures. Treatment commonly involves the use of continuous lumbar drainage. A retrospective chart review of 32 consecutive patients who required placement of lumbar drain by the otorhinolaryngology and neurosurgical services from March 1988 through July 1991 was undertaken to assess possible complications. The complications found were readily separated into minor and major categories on the basis of the possibility of permanent morbidity or mortality. Minor complications, including subjective complaints of headache, nausea, vomiting, etc., were noted in 59% of patients. Major complications were observed in four of 32 patients (12.5%), including unilateral occlusion of the posterior cerebral artery and unilateral true vocal cord paralysis. Alleviation of all complications was achieved by cessation of lumbar drainage. These cases are presented with discussion of pathogenesis. These findings demonstrate the possibility of potentially serious complications that mandate close monitoring of patients who require continuous lumbar drainage.


Meditation and the startle response: a case study.
The effects of two kinds of meditation (open presence and focused) on the facial and physiological aspects of the defensive response to an aversive startle stimulus were studied in a Buddhist monk with approximately 40 years of meditation experience. The participant was exposed to a 115-db, 100-ms acoustic startle stimulus under the 2 meditation conditions, a distraction condition (to control for cognitive and attentional load) and an unanticipated condition (startle presented without warning or instruction). A completely counterbalanced 24-trial, single-subject design was used, with each condition repeated 6 times. Most aspects of the participant's responses in the unanticipated condition did not differ from those of a comparison group of 12 age-matched male controls. Both kinds of meditation produced physiological and facial responses to the startle that were smaller than in the distraction condition. Within meditation conditions, open presence meditation produced smaller physiological and facial responses than focused meditation. These results from a single highly expert meditator indicate that these 2 kinds of meditation can differentially alter the magnitude of a primitive defensive response.


Sodium transport in plant cells.
Salinity limits plant growth and impairs agricultural productivity. There is a wide spectrum of plant responses to salinity that are defined by a range of adaptations at the cellular and the whole-plant levels, however, the mechanisms of sodium transport appear to be fundamentally similar. At the cellular level, sodium ions gain entry via several plasma membrane channels. As cytoplasmic sodium is toxic above threshold levels, it is extruded by plasma membrane Na(+)/H(+) antiports that are energized by the proton gradient generated by the plasma membrane ATPase. Cytoplasmic Na(+) may also be compartmentalized by vacuolar Na(+)/H(+) antiports. These transporters are energized by the proton gradient generated by the vacuolar H(+)-ATPase and H(+)-PPiase. Here, the mechanisms of sodium entry, extrusion, and compartmentation are reviewed, with a discussion of recent progress on the cloning and characterization, directly in planta and in yeast, of some of the proteins involved in sodium transport.


ZMYND8 is a master regulator of 27-hydroxycholesterol that promotes tumorigenicity of breast cancer stem cells.
27-Hydroxycholesterol (27-HC) is the most abundant oxysterol that increases the risk of breast cancer progression. However, little is known about epigenetic regulation of 27-HC metabolism and its role in breast tumor initiation. Using genetic mouse mammary tumor and human breast cancer models, we showed here that the histone reader ZMYND8 was selectively expressed in breast cancer stem cells (BCSCs) and promoted epithelial-mesenchymal transition (EMT), BCSC maintenance and self-renewal, and oncogenic transformation through its epigenetic functions, leading to breast tumor initiation. Mechanistically, ZMYND8 was a master transcriptional regulator of 27-HC metabolism. It increased cholesterol biosynthesis and oxidation but blocked cholesterol efflux and 27-HC catabolism, leading to accumulation of 27-HC in BCSCs. Consequently, 27-HC promoted EMT, oncogenic transformation, and tumor initiation through activation of liver X receptor. These findings reveal that ZMYND8 is an epigenetic booster that drives breast tumor initiation through metabolic reprogramming.


Muscarinic acetylcholine receptor activation increases transcellular transport of macromolecules across mouse and human intestinal epithelium in vitro.
The intestinal epithelium acts as a barrier restricting uptake of luminal macromolecules such as dietary antigens and microbes. Here, we examined the role of cholinergic signalling in the regulation of permeability to macromolecules. Mouse jejunum was mounted in Ussing chambers and permeability was determined by measuring the flux of the antigen-sized protein, horseradish peroxidase (HRP), across the tissue. Baseline HRP permeability was significantly reduced by neural blockade with tetrodotoxin or cholinergic muscarinic antagonism with atropine, suggesting that ongoing release of endogenous acetylcholine from enteric nerves regulates barrier function. Exogenous addition of the muscarinic agonist bethanechol caused significant increases in both HRP flux and the area of HRP-containing endosomes in enterocytes. Bethanechol-enhanced HRP flux was abrogated by the M3 receptor antagonist, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), the phospholipase A(2) inhibitor quinacrine, and the cyclooxygenase inhibitor indomethacin. Complementary in vitro studies showed direct effects of bethanechol on T84 epithelial cells, where increased HRP uptake was associated with increased F-actin, and increased cytosolic phospholipase A(2) (cPLA(2)) phosphorylation. Taken together, these results provide evidence for cholinergic regulation of transepithelial transport of macromolecules, mainly mediated by activation of M3 receptors with subsequent involvement of phospholipase A(2) and cyclooxygenase products.


Anti-RBD IgG antibodies and neutralizing antibody levels after the second BNT162b2 dose in patients with plasma cell disorders.
Patients with plasma cell disorders (PCD) are at an increased risk for severe morbidity and mortality due to COVID-19. Recent data have suggested that patients with hematological malignancies, including those with PCD, have suboptimal antibody response to COVID-19 vaccination. We compared the antibody titers of 213 patients with PCD to those of 213 immunocompetent healthcare workers after the second vaccine dose of the BNT162b2 mRNA vaccine. Blood samples were taken 2-4 weeks after the second vaccination and analyzed for anti-receptor binding-domain immunoglobulin G (RBD-IgG) antibodies and neutralizing antibodies (NA). At a median of 20 days after the second vaccine dose, 172 patients (80.8%) developed anti-RBD-IgG antibodies with a geometric mean titer (GMT) of 2.7 (95% confidence interval [CI], 2.4-3.1). In the control group 210 (98.9%) developed anti-RBD-IgG antibodies after a median of 21 days, with a GMT of 5.17 (95%CI, 4.8-5.6), p<0.0001. NA were observed in 151 patients with MM (70.9%) and in 210 controls (98.9%). The GMT of NA in patients with MM and controls was 84.4 (95% CI, 59.0-120.6), and 420.2 (95% CI, 341.4-517.1), respectively (p<0.0001). Multivariable logistic regression revealed that the number of prior therapy lines and age were significant predictors of poor humoral response among patients with MM. Injection site reaction, headache and fatigue were the most common adverse events after vaccination. Adverse events were less common in patients with MM than in controls. In conclusion, a significant percentage of patients with MM developed protecting NA to the BNT162b2 mRNA vaccine, which appears to be safe in this patient population.


Modelling the physics in the iterative reconstruction for transmission computed tomography.
There is an increasing interest in iterative reconstruction (IR) as a key tool to improve quality and increase applicability of x-ray CT imaging. IR has the ability to significantly reduce patient dose; it provides the flexibility to reconstruct images from arbitrary x-ray system geometries and allows one to include detailed models of photon transport and detection physics to accurately correct for a wide variety of image degrading effects. This paper reviews discretization issues and modelling of finite spatial resolution, Compton scatter in the scanned object, data noise and the energy spectrum. The widespread implementation of IR with a highly accurate model-based correction, however, still requires significant effort. In addition, new hardware will provide new opportunities and challenges to improve CT with new modelling.


Genetic determinants of weight of fast- and slow-twitch skeletal muscles in old mice.
The main goal of the study was to explore the genetic architecture underlying muscle weight in old mice. Weight of soleus, tibialis anterior (TA), extensor digitorum longus (EDL), and gastrocnemius muscles was measured in the C57BL/6J (B6) and DBA/2J (D2) strains and derivative generations: a panel of the BXD recombinant inbred (RI) strains and a B6D2 F(2) intercross at the age of 800 days. The between-strain difference in muscle weight (B6 > D2) ranged between 16% and 38%. Linkage analysis identified suggestive quantitative trait loci (QTL) on Chromosomes (Chr) 2, 6, 7, 8, 19, and X that influenced muscle weight in the 800-day-old group. Comparison of weights at 200, 500, and 800 days revealed a variable effect of age among the four muscles. Linkage analysis in the B6D2 F(2) population combined across the three different age groups identified muscle-, sex-, and age-specific QTL on Chr 1, 2, 3, 5, 6, 8, 9, 11, 13, 17, X, and Y. Genetic factors that influence the rate of weight change (within-strain weight difference at two ages) over the lifespan of BXD RIs were mapped to the markers D2Mit369 and D3Mit130 at the genome-wide p < 0.05 for TA muscle in males (between 200 and 800 days) and females (between 500 and 800 days), respectively. Analysis of all age groups supported previous findings that the genetic effects may be muscle-, age-, and sex-specific.


Introduction of customized inserts for s-treamlined assembly and optimization of BioBrick synthetic genetic circuits.
BioBrick standard biological parts are designed to make biological systems easier to engineer (e.g. assemble, manipulate, and modify). There are over 5,000 parts available in the Registry of Standard Biological Parts that can be easily assembled into genetic circuits using a standard assembly technique. The standardization of the assembly technique has allowed for wide distribution to a large number of users -- the parts are reusable and interchangeable during the assembly process. The standard assembly process, however, has some limitations. In particular it does not allow for modification of already assembled biological circuits, addition of protein tags to pre-existing BioBrick parts, or addition of non-BioBrick parts to assemblies. In this paper we describe a simple technique for rapid generation of synthetic biological circuits using introduction of customized inserts. We demonstrate its use in Escherichia coli (E. coli) to express green fluorescent protein (GFP) at pre-calculated relative levels and to add an N-terminal tag to GFP. The technique uses a new BioBrick part (called a BioScaffold) that can be inserted into cloning vectors and excised from them to leave a gap into which other DNA elements can be placed. The removal of the BioScaffold is performed by a Type IIB restriction enzyme (REase) that recognizes the BioScaffold but cuts into the surrounding sequences; therefore, the placement and removal of the BioScaffold allows the creation of seamless connections between arbitrary DNA sequences in cloning vectors. The BioScaffold contains a built-in red fluorescent protein (RFP) reporter; successful insertion of the BioScaffold is, thus, accompanied by gain of red fluorescence and its removal is manifested by disappearance of the red fluorescence. The ability to perform targeted modifications of existing BioBrick circuits with BioScaffolds (1) simplifies and speeds up the iterative design-build-test process through direct reuse of existing circuits, (2) allows incorporation of sequences incompatible with BioBrick assembly into BioBrick circuits (3) removes scar sequences between standard biological parts, and (4) provides a route to adapt synthetic biology innovations to BioBrick assembly through the creation of new parts rather than new assembly standards or parts collections.


Decisions behind career choice for nurse practitioners: independent versus collaborative practice and motivational-needs behavior.
Nurse practitioners (NPs) can take an active role in defining and establishing their careers. Prepared as advanced practice nurses with specific assessment skills, primary care NPs have the opportunity to become independent or collaborative practitioners. This report examines the published work in the area of practice choice and motivational-needs behavior. Interviews with collaborative and independent primary care NPs were conducted. Against the framework of well-established personality testing methods, open-ended interview questions were developed to elicit specific motivational-needs-based behavior characteristics. The motivational needs examined included the need for achievement, power, and affiliation. The interview findings were then synthesized using needs-based behavior theory. This new platform for role decision ultimately can prepare NPs to make informed career choices.


Diazepam-induced loss of inhibitory synapses mediated by PLCδ/ Ca2+/calcineurin signalling downstream of GABAA receptors.
Benzodiazepines facilitate the inhibitory actions of GABA by binding to γ-aminobutyric acid type A receptors (GABAARs), GABA-gated chloride/bicarbonate channels, which are the key mediators of transmission at inhibitory synapses in the brain. This activity underpins potent anxiolytic, anticonvulsant and hypnotic effects of benzodiazepines in patients. However, extended benzodiazepine treatments lead to development of tolerance, a process which, despite its important therapeutic implications, remains poorly characterised. Here we report that prolonged exposure to diazepam, the most widely used benzodiazepine in clinic, leads to a gradual disruption of neuronal inhibitory GABAergic synapses. The loss of synapses and the preceding, time- and dose-dependent decrease in surface levels of GABAARs, mediated by dynamin-dependent internalisation, were blocked by Ro 15-1788, a competitive benzodiazepine antagonist, and bicuculline, a competitive GABA antagonist, indicating that prolonged enhancement of GABAAR activity by diazepam is integral to the underlying molecular mechanism. Characterisation of this mechanism has revealed a metabotropic-type signalling downstream of GABAARs, involving mobilisation of Ca2+ from the intracellular stores and activation of the Ca2+/calmodulin-dependent phosphatase calcineurin, which, in turn, dephosphorylates GABAARs and promotes their endocytosis, leading to disassembly of inhibitory synapses. Furthermore, functional coupling between GABAARs and Ca2+ stores was sensitive to phospholipase C (PLC) inhibition by U73122, and regulated by PLCδ, a PLC isoform found in direct association with GABAARs. Thus, a PLCδ/Ca2+/calcineurin signalling cascade converts the initial enhancement of GABAARs by benzodiazepines to a long-term downregulation of GABAergic synapses, this potentially underpinning the development of pharmacological and behavioural tolerance to these widely prescribed drugs.


Contextual attributes to promote positive social interdependence in problem-based learning: a focus group study.
Problem-based learning (PBL) is classified as a collaborative learning approach, wherein students learn while contributing meaning to experiences and interactions with others. An important theoretical fundament of PBL is social interdependence theory (SIT) because positive social interdependence within a group has been found to be key to better learning performance and future attitudes towards team practice. However, most previous studies in health professions education focused on cognitive outcomes, and few studies have focused on collaborative behaviors in PBL groups. The lack of this empirical insight makes implementation of PBL difficult, especially in contexts where there is limited experience with collaborative learning. Therefore, the aim of this study was to elucidate what promotes or hinders positive social interdependence and how the attributes work during PBL. We conducted four focus groups among clinical year medical students (n = 26) who participated in PBL tutorials in the formal curriculum. We asked semi-structured questions that corresponded with the overall concept of SIT. We analyzed the transcript using constructivist grounded theory and developed a model to explain contextual attributes that promote or hinder positive social interdependence in PBL. Two contextual attributes of "academic inquisition" and "desire for efficiency" affect social interdependence among a student group in PBL. Academic inquisition is students' desire to engage in their academic learning, and desire for efficiency is students' attitude toward learning as an imposed duty and desire to complete it as quickly as possible. These attributes are initially mutually conflicting and constructing social interdependence through multiple steps including inquisition from a case, seeking efficient work, sharing interest in problem solving, expecting mutual contributions, and complementing learning objectives. These findings will contribute to understanding collaborative learning environments in PBL and may help explain contexts where PBL is less successful. The model can also be used as a tool to support innovation of PBL as collaborative learning.


Functional MOF-Based Materials for Environmental and Biomedical Applications: A Critical Review.
Over the last ten years, there has been a growing interest in metal-organic frameworks (MOFs), which are a unique category of porous materials that combine organic and inorganic components. MOFs have garnered significant attention due to their highly favorable characteristics, such as environmentally friendly nature, enhanced surface area and pore volume, hierarchical arrangements, and adjustable properties, as well as their versatile applications in fields such as chemical engineering, materials science, and the environmental and biomedical sectors. This article centers on examining the advancements in using MOFs for environmental remediation purposes. Additionally, it discusses the latest developments in employing MOFs as potential tools for disease diagnosis and drug delivery across various ailments, including cancer, diabetes, neurological disorders, and ocular diseases. Firstly, a concise overview of MOF evolution and the synthetic techniques employed for creating MOFs are provided, presenting their advantages and limitations. Subsequently, the challenges, potential avenues, and perspectives for future advancements in the utilization of MOFs in the respective application domains are addressed. Lastly, a comprehensive comparison of the materials presently employed in these applications is conducted.


Marburg and Ebola viruses as aerosol threats.
Ebola and Marburg viruses are the sole members of the genus Filovirus in the family Filoviridae. There has been considerable media attention and fear generated by outbreaks of filoviruses because they can cause a severe viral hemorrhagic fever (VHF) syndrome that has a rapid onset and high mortality. Although they are not naturally transmitted by aerosol, they are highly infectious as respirable particles under laboratory conditions. For these and other reasons, filoviruses are classified as category A biological weapons. However, there is very little data from animal studies with aerosolized filoviruses. Animal models of filovirus exposure are not well characterized, and there are discrepancies between these models and what has been observed in human outbreaks. Building on published results from aerosol studies, as well as a review of the history, epidemiology, and disease course of naturally occurring outbreaks, we offer an aerobiologist's perspective on the threat posed by aerosolized filoviruses.


The impact of the HIRA histone chaperone upon global nucleosome architecture.
HIRA is an evolutionarily conserved histone chaperone that mediates replication-independent nucleosome assembly and is important for a variety of processes such as cell cycle progression, development, and senescence. Here we have used a chromatin sequencing approach to determine the genome-wide contribution of HIRA to nucleosome organization in Schizosaccharomyces pombe. Cells lacking HIRA experience a global reduction in nucleosome occupancy at gene sequences, consistent with the proposed role for HIRA in chromatin reassembly behind elongating RNA polymerase II. In addition, we find that at its target promoters, HIRA commonly maintains the full occupancy of the -1 nucleosome. HIRA does not affect global chromatin structure at replication origins or in rDNA repeats but is required for nucleosome occupancy in silent regions of the genome. Nucleosome organization associated with the heterochromatic (dg-dh) repeats located at the centromere is perturbed by loss of HIRA function and furthermore HIRA is required for normal nucleosome occupancy at Tf2 LTR retrotransposons. Overall, our data indicate that HIRA plays an important role in maintaining nucleosome architecture at both euchromatic and heterochromatic loci.


[Minilaparotomy in surgical treatment of calculous cholecystitis].
A set of instruments devised by the CAN company was used in performing cholecystectomy in 15 patients. It proved adequate for the operative situation and allowed surgical manipulations to be carried out through a 30-50 mm minilaparotomy. Cases in which the approach had to be widened or intraoperative complications occurred were not encountered. The indices of external respiration function and the volumetric parameters decrease a little but are rapidly restored during the postoperative period. This confirms the mild traumatic character of the operation and is consistent with the absence of pulmonary complications and negligible manifestation of postoperative intestinal paresis.


The Development of a Long Peripheral Catheter Program at a Large Pediatric Academic Center: A Pilot Study.
The purpose of this study was to evaluate the feasibility of a new long peripheral catheter (LPC) program at a large academic center in an effort to reduce the use of peripherally inserted central catheters (PICCs) and their related complications. The pilot participants were hospitalized children, age >2 years, with a need for noncentral intravenous access for 2 to 29 days, or laboratory blood draw >5 times per day. Patients expected to discharge with intravenous access were excluded. Included in the pilot program development were a literature review, 1-year baseline data analysis, and program design and implementation. A multidisciplinary committee developed and implemented the program from December 2018 to September 2019. LPCs were placed from August to September 2019. Regarding the baseline data, between July 2018 and June 2019, 584 PICCs were placed in 461 patients. Of these, 139 PICCs (24%) did not meet requirements necessitating central access and, potentially, could have been avoided if an LPC alternative were available at the time. For the LPC pilot program, 20 LPCs were placed in 19 patients. The median age was 11 (interquartile range of 7-15). The insertion success rate was 83%. There were no serious complications, such as venous thrombosis or catheter-related bloodstream infection. The total rate of minor complications was 35%: the rate of occlusions was 10% (n = 2), and the rate of dislodgement was 25% (n = 5). The catheter failure rate was 74 per 1000 catheter-days. The mean line duration was 6 days. There is a role for LPCs in hospitalized children requiring durable vascular access. Multispecialty designed pilot implementation of an LPC program was successful at an academic pediatric hospital.


Liquid-Infused Surfaces: A Review of Theory, Design, and Applications.
Due to inspiration from the Nepenthes pitcher plant, a frontier of devices has emerged with unmatched capabilities. Liquid-infused surfaces (LISs), particularly known for their liquid-repelling behavior under low tilting angles (<5°), have demonstrated a plethora of applications in medical, marine, energy, industrial, and environmental materials. This review presents recent developments of LIS technology and its prospective to define the future direction of this technology in solving tomorrow's real-life challenges. First, an introduction to the different models explaining the physical phenomena of these surfaces, their wettability, and viscous-dependent frictional forces is discussed. Then, an outline of different emerging strategies required to fabricate a stable liquid-infused interface is presented, including different substrates, lubricants, surface chemistries, and design parameters which can be tuned depending on the application. Furthermore, applications of LIS coatings in the areas of anticorrosion, antifouling, anti-icing, self-healing, droplet manipulation, and biomedical devices will be presented followed by the limitations and future direction of this technology.


Terebratulide brachiopod shell biomineralization by mantle epithelial cells.
To understand mineral transport pathways for shell secretion and to assess differences in cellular activity during mineralization, we imaged with TEM and FE-SEM ultrastructural characteristics of outer mantle epithelium (OME) cells. Imaging was carried out on Magellania venosa shells embedded/etched, chemically fixed/decalcified and high-pressure frozen/freeze-substituted samples from the commissure, central shell portions and from puncta. Imaging results are complemented with morphometric evaluations of volume fractions of membrane-bound organelles. At the commissure the OME consists of several layers of cells. These cells form oblique extensions that, in cross-section, are round below the primary layer and flat underneath fibres. At the commissure the OME is multi-cell layered, in central shell regions it is single-cell layered. When actively secreting shell carbonate extrapallial space is lacking, because OME cells are in direct contact with the calcite of the forming fibres. Upon termination of secretion, OME cells attach via apical hemidesmosomes to extracellular matrix membranes that line the proximal surface of fibres. At the commissure volume fractions for vesicles, mitochondria and lysosomes are higher relative to single-cell layered regions, whereas for endoplasmic-reticulum and Golgi apparatus there is no difference. FE-SEM, TEM imaging reveals the lack of extrapallial space between OME cells and developing fibres. In addition, there is no indication for an amorphous precursor within fibres when these are in active secretion mode. Accordingly, our results do not support transport of minerals by vesicles from cells to sites of mineralization, rather by transfer of carbonate ions via transport mechanisms associated with OME cell membranes.


Implications of urine F2-isoprostane metabolite concentration in horses with colic and its potential use as a predictor for surgical intervention.
F2-isoprostanes have been used extensively to quantify lipid peroxidation in association with risk factors in various diseases. Horses with colic may have intestinal ischaemia and/or inflammation characterised by oxidative stress and increased production of isoprostanes. To gather preliminary data regarding the feasibility of using urine F2-isoprostanes and isoprostane metabolites as early screening tools for the presence of gastrointestinal disease requiring surgical intervention in horses and ultimately develop a stall-side test capable of identifying these horses as early as possible for timely referral. Concentrations of urine isoprostane and isoprostane metabolite were determined by mass spectroscopy and normalised to urine creatinine (Cr) concentrations in urine samples from 42 healthy control horses and 43 horses with gastrointestinal pain or colic. Horses with colic were treated medically (n = 21) or surgically (n = 22). Mean ± s.d. concentrations of urine isoprostane and isoprostane metabolite were significantly higher in horses with colic (2.94 ± 1.69 and 0.31 ± 0.22 ng/mg Cr, respectively), compared to control horses (1.89 ± 1.39 and 0.22 ± 0.08 ng/mg Cr, respectively). Mean urine isoprostane metabolite concentrations were significantly higher in horses undergoing surgery (0.38 ± 0.28 ng/mg Cr) compared to controls and medical colics (0.26 ± 0.11 ng/mg Cr). Nonsurvivors had significantly higher mean urine isoprostane metabolite concentrations (0.47 ± 0.39 ng/mg Cr) than control or surviving colic horses (0.29 ± 0.24 ng/mg Cr). Measurement of urine isoprostane metabolite concentration may be a useful prognostic indicator in equine colic. Urine isoprostane metabolites may aid in early recognition of surgical colic. Isoprostanes are a potential therapeutic target to prevent further systemic and gastrointestinal tissue injury in horses with colic.


High brightness single mode source of correlated photon pairs using a photonic crystal fiber.
We demonstrate a picosecond source of correlated photon pairs using a micro-structured fibre with zero dispersion around 715 nm wavelength. The fibre is pumped in the normal dispersion regime at ~708 nm and phase matching is satisfied for widely spaced parametric wavelengths. Here we generate up to 10;7 photon pairs per second in the fibre at wavelengths of 587 nm and 897 nm, while on collecting this light in single-mode-fibre-coupled Silicon avalanche diode photon counting detectors, we detect ~3.2x10;5 coincidences per second at pump power 0.5 mW.


Comparative retinal anatomy in four species of elasmobranch.
Using both light and transmission electron microscopy, we examined the retinal anatomy of four elasmobranch species with differing ecologies: the bull shark Carcharhinus leucas, Port Jackson shark Heterodontus portusjacksoni, epaulette shark Hemiscyllium ocellatum and pink whipray Himantura fai. Their retinas are typical of other vertebrates, having three nuclear and two synaptic layers, but are characterised by very large horizontal cells, low densities of ganglion cells (many of which are displaced to the inner nuclear and inner plexiform layers) and the presence of numerous myelinated axons within the nerve fibre layer. Carcharhinus leucas, H. fai and H. ocellatum have duplex retinas containing both rods and single cones. The peak ratio of rods to cones is much lower in C. leucas (4:1) and H. fai (3:1) compared to H. ocellatum (19:1), reflecting differences in diel activity patterns. No cones were observed in the retina of H. portusjacksoni, which is strongly nocturnal. The cones of H. fai lack a distinct myoid and their nuclei are located in a discrete layer sclerad to the external limiting membrane (ELM), whereas those of C. leucas and H. ocellatum have an obvious myoid, and their nuclei are located vitread to the ELM. No double/twin cones were observed in any species. Incorporating data from other studies, there is a clear correlation between rod outer segment volume and visual ecology in elasmobranchs, with smaller volumes found in partly diurnal pelagic species and larger volumes in benthic nocturnal species. This trend may reflect fundamental differences in visual temporal resolution between active and more sedentary species.


Photophysical Properties of Organoplatinum(II) Compounds and Derived Self-Assembled Metallacycles and Metallacages: Fluorescence and its Applications.
Over the past couple of decades, coordination-driven self-assembly has evolved as a broad multidisciplinary domain that not only covers the syntheses of aesthetically pleasing supramolecular architectures but also emerges as a method to form new optical materials, chemical sensors, theranostic agents, and compounds with light-harvesting and emissive properties. The majority of these applications depend upon investigations that reveal the photophysical nature and electronic structure of supramolecular coordination complexes (SCCs), including two-dimensional (2D) metallacycles and three-dimensional (3D) metallacages. As such, well-defined absorption and emission spectra are important for a given SCC to be used for sensing, bioimaging, and other applications with molecular fluorescence being an important component. In this Account, we summarize the photophysical properties of some bis(phosphine)organoplatinum(II) compounds and their discrete SCCs. The platinum(II) based organometallic precursors typically display spectral red-shifts and have low fluorescence quantum yields and short fluorescence lifetimes compared to their organic counterparts because the introduction of metal centers enhances both intersystem crossing (ISC) and intramolecular charge transfer (ICT) processes, which can compete with the fluorescence emissions. Likewise ligands with conjugation can also increase the ICT process; hence the corresponding organoplatinum(II) compounds undergo a further decrease in fluorescence lifetimes. The use of endohedral amine functionalized 120°-bispyridyl ligands can dramatically enhance the emission properties of the resultant organoplatinum(II) based SCCs. As such these SCCs display emissions in the visible region (ca. 400-500 nm) and are significantly red-shifted (ca. 80-100 nm) compared to the ligands. This key feature makes them suitable as supramolecular theranostic agents wherein these unique emission properties provide diagnostic spectroscopic handles and the organoplatinum(II) centers act as potential anticancer agents. Using steady state and time-resolved-spectroscopic techniques and quantum computations in concert, we have determined that the emissive properties stem from the ligand-centered transitions involving π-type molecular orbitals with modest contributions from the metal-based orbitals. The self-assembly and the photophysics of organoplatinum(II) ← 3-substituted pyridyl based SCCs are highly diverse. Subtle changes in the ligands' structures can form molecular congener systems with distinct conformational and photophysical properties. Furthermore, the heterometallic SCCs described herein possess rich photophysical properties and can be used for sensing based applications. Tetraphenylethylene (TPE) based SCCs display emissions in the aggregated state as well as in dilute solutions. This is a unique phenomenon that bridges the aggregation caused quenching (ACQ) and aggregation induced emission (AIE) effects. Moreover, a TPE based metallacage exhibits solvatoluminescence, including white light emission in THF solvent, and can act as a fluorescence-sensor for structurally similar ester compounds.


[Inequality and damaged teeth: oral sequelae from living in poverty in the Dendê community, Fortaleza, Ceará, Brazil].
This anthropological study focuses on the experience of dental disease in the context of poverty in Northeast Brazil. For six months in 2004, ethnographic interviews, narratives, and participatory observation were conducted with 31 residents of the low-income Dendê neighborhood in Fortaleza, Ceará, and the results were analyzed using a hermeneutic-dialectic method. Precarious living conditions make healthcare a difficult priority. Even when individuals experience acute toothache, seeking dental care is perceived as a "luxury" rather than a citizen's right. Difficult access to services and poor-quality restorations favor tooth extraction as the most effective intervention. Local residents complain about their deteriorating oral health and seek help from substandard clinics, patronizing politicians, and traditional healers. In this context, dental disease differs according to social class, leaves the sequelae of inequality, jeopardizes self-esteem, and hinders social inclusion. Dealing with such inequality requires a deeper understanding of the social determinants of health, reducing injustices in access to quality care, eliminating stigma, and empowering community members to confront the structural forces affecting their lives.


GABA concentration in posterior cingulate cortex predicts putamen response during resting state fMRI.
The role of neurotransmitters in the activity of resting state networks has been gaining attention and has become a field of research with magnetic resonance spectroscopy (MRS) being one of the key techniques. MRS permits the measurement of γ-aminobutyric acid (GABA) and glutamate levels, the central biochemical constituents of the excitation-inhibition balance in vivo. The inhibitory effects of GABA in the brain have been largely investigated in relation to the activity of resting state networks in functional magnetic resonance imaging (fMRI). In this study GABA concentration in the posterior cingulate cortex (PCC) was measured using single voxel spectra acquired with standard point resolved spectroscopy (PRESS) from 20 healthy male volunteers at 3 T. Resting state fMRI was consecutively measured and the values of GABA/Creatine+Phosphocreatine ratio (GABA ratio) were included in a general linear model matrix as a step of dual regression analysis in order to identify voxels whose neuroimaging metrics during rest were related to individual levels of the GABA ratio. Our data show that the connection strength of putamen to the default-mode network during resting state has a negative linear relationship with the GABA ratio measured in the PCC. These findings highlight the role of PCC and GABA in segregation of the motor input, which is an inherent condition that characterises resting state.


Management of protein intake in the fruit fly Anastrepha fraterculus.
This work tested if carbohydrates and proteins ingestion is regulated in the South American fruit fly, Anastrepha fraterculus, to optimize survival and reproduction. Adult food treatments were established by providing sugar and hydrolyzed yeast in various combinations either alone or mixed at a standard 3:1 ratio (sugar:hydrolyzed yeast). Individual food consumption was assessed and related to survival patterns. The effects of adult feeding on fecundity and fertility patterns were investigated in groups of flies. Sugar consumption was the lowest in the treatment where it was provided with hydrolyzed yeast at a fixed 3:1 ratio. Consumption of hydrolyzed yeast did not differ between this treatment and the one in which this solution was complemented with one solution of sugar. It seems that a mixture of sugar and hydrolyzed yeast at a fixed ratio of 3:1, respectively, restricts extra ingestion of sugar; most probably because of negative response of the fly to overconsumption of protein. Survival was affected by the treatments, being lower in those cases where protein was at the fixed ratio. Group experiments revealed that protein restriction expanded longevity and decreased egg production. In contrast, egg production was enhanced when flies were kept continuously with a mixture of yeast and sugar plus an extra source of sugar, and this was not in detriment of survival. Our results suggest that fixed sugar-protein ratios in which protein is in excess affects fitness components such as longevity and reproduction. These findings are discussed from a theoretical and applied perspective in the context of pest control by means of the sterile insect technique.


The feasibility of clinical endpoint trials in HIV infection in the highly active antiretroviral treatment (HAART) era.
Planning clinical-endpoint trials in patients with HIV remain difficult as long-term follow-up of many patients is required. Cohort studies of patients with HIV can provide key estimates of the likely disease progression, required sample size and follow-up. To verify the assumptions used in designing ESPRIT, a large randomized clinical trial assessing the clinical benefit of interleukin-2 treatment in patients with HIV infection, to use EuroSIDA to mimic the inclusion criterion of ESPRIT in order to compare the observed event rate in ESPRIT with the projected rate in EuroSIDA, and to project the required length of ESPRIT. Patients in EuroSIDA who satisfied the ESPRIT recruitment criteria were selected. Patients were followed from baseline to new AIDS or death. The incidence of clinical progression in the selected EuroSIDA patients (N = 4482) was 1.5 per 100 PYFU (95% CI 1.3-1.7), and did not increase with increasing time from baseline, contrary to what was assumed in the design of the ESPRIT trial. In ESPRIT (N = 4150), for which the comparative data remain blinded, the incidence was 1.1 per 100 PYFU (95% CI 0.9-1.3), with no increase over time. The average follow-up required to complete ESPRIT and accrue the 320 events required by protocol would be seven years, 10 months using the projected rates from the EuroSIDA study, and seven years, 11 months if the observed event rate in ESPRIT continued unchanged. Differences between patients recruited to observational studies or clinical trials cannot always be adjusted for. Event rates in EuroSIDA were similar in the first two years to those used in the design of ESPRIT, but did not increase over time, leading to an increase in the expected duration of ESPRIT. Clinical endpoint trials in HIV infection remain feasible, and large cohort studies are critical to the planning and ongoing assessment of design assumptions in such trials. The underlying assumptions of the clinical trial should be re-examined to ensure the original trial assumptions remain valid.


Mental health and environmental factors in adults: A population-based network analysis.
Few studies have assessed the multifactorial nature of environmental influences on population mental health. In this large-scale, population-based study of adults, we applied network analysis to study the relationship between environmental factors and symptoms of depression, anxiety, and well-being. We estimated networks with overall mental health nodes and individual symptoms to assess both broad and fine-grained associations between environmental factors and mental health. Finally, we conducted an out-of-sample replication in an independent large-scale sample to assess the robustness of our results. Across 31,000 adults randomly sampled from the Norwegian population, we identified associations between numerous environmental characteristics and mental health. Recent discrimination and unsupportive social environments were strongly associated with lower population well-being and higher levels of mental illness symptoms, respectively. The most strongly connected variables in the networks were environmental factors, including perceived problems with crime, violence, or vandalism in the residential area, worrying about violence or threats when outside, and problems with noise or contamination at home. Substantial variation in population mental health was explained by environmental factors included in the networks. Replicability of the results was excellent and suggestive of strong robustness of the results across samples. Our findings are indicative of the importance of environmental factors, such as the social environment, housing satisfaction, and residential area characteristics, for multiple aspects of population mental health. We identify several environmental factors that represent potentially useful targets for future studies and public health efforts seeking to improve mental health in the general population. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


Semantically Transparent and Opaque Compounds in German Noun-Phrase Production: Evidence for Morphemes in Speaking.
This study examines the lexical representation and processing of noun-noun compounds and their grammatical gender during speech production in German, a language that codes for grammatical gender (masculine, feminine, and neuter). Using a picture-word interference paradigm, participants produced determiner-compound noun phrases in response to pictures, while ignoring written distractor words. Compound targets were either semantically transparent (e.g., birdhouse) or opaque (e.g., hotdog), and their constituent nouns either had the same or a different gender (internal gender match). Effects of gender-congruent but otherwise unrelated distractor nouns, and of two morphologically related distractors corresponding to the first or second constituent were assessed relative to a completely unrelated, gender-incongruent distractor baseline. Both constituent distractors strongly facilitated compound naming, and these effects were independent of the targets' semantic transparency. This supports retrieval of constituent morphemes for semantically transparent and opaque compounds during speech production. Furthermore, gender congruency between compounds and distractors did not speed up naming in general, but interacted with gender match of the compounds' constituent nouns, and their semantic transparency. A significant gender-congruency effect was obtained with semantically transparent compounds, consisting of two constituent nouns of the same gender, only. In principle, this pattern is compatible with a multiple lemma representation account for semantically transparent, but not for opaque compounds. The data also fit with a more parsimonious, holistic representation for all compounds at the lemma level, when differences in co-activation patterns for semantically transparent and opaque compounds are considered.


Study protocol: Asking QUestions about Alcohol in pregnancy (AQUA): a longitudinal cohort study of fetal effects of low to moderate alcohol exposure.
Despite extensive research, a direct correlation between low to moderate prenatal alcohol exposure (PAE) and Fetal Alcohol Spectrum Disorders has been elusive. Conflicting results are attributed to a lack of accurate and detailed data on PAE and incomplete information on contributing factors. The public health effectiveness of policies recommending complete abstinence from alcohol during pregnancy is challenged by the high frequency of unplanned pregnancies, where many women consumed some alcohol prior to pregnancy recognition. There is a need for research evidence emphasizing timing and dosage of PAE and its effects on child development. Asking QUestions about Alcohol (AQUA) is a longitudinal cohort aiming to clarify the complex effects of low to moderate PAE using specifically developed and tested questions incorporating dose, pattern and timing of exposure. From 2011, 2146 pregnant women completed a questionnaire at 8-18 weeks of pregnancy. Further prenatal data collection took place via a questionnaire at 26-28 weeks and 35 weeks gestation. Extensive information was obtained on a large number of risk factors to assist in understanding the heterogeneous nature of PAE effects. 1571 women (73%) completed all three pregnancy questionnaires. A biobank of DNA from maternal and infant buccal cells, placental biopsies and cord blood mononuclear cells will be used to examine epigenetic state at birth as well as genetic factors in the mother and child. Participants will be followed up at 12 and 24 months after birth to assess child health and measure infant behavioural and sensory difficulties, as well as family environment and parenting styles. A subgroup of the cohort will have 3D facial photography of their child at 12 months and a comprehensive developmental assessment (Bayley Scales of Infant & Toddler Development, Bayley-III) at two years of age. Using detailed, prospective methods of data collection, the AQUA study will comprehensively examine the effects of low to moderate alcohol consumption throughout pregnancy on child health and development, including the role of key mediators and confounders. These data will ultimately contribute to policy review and development, health professional education and information about alcohol consumption for pregnant women in the future.


Multiplex gene regulation: a two-tiered approach to transgene regulation in transgenic mice.
Transgenic mice have been used to study gene function and regulation by introducing inducible or tissue-specific transgenes. This approach is generally limited to studying gene function in adult mice since ectopic expression of many interesting genes is disease causing or may be lethal to the developing embryo. To extend the utility of the transgenic mouse system to the early stages of embryogenesis, we have developed a two-tiered method of gene regulation to control transgene expression. Our multiplex gene regulatory system (MGR) allows the establishment of transgenic lines that harbor inducible potentially lethal transgenes. These inducible transgenes are activated only when mated to a second transgenic animal. Induction in the MGR system provides a high degree of temporal and spatial control over transgene expression and should be suitable for engineering "gain of function mutations" for many developmental genes.


Electrospun Decellularized Lung Matrix Scaffold for Airway Smooth Muscle Culture.
Chronic respiratory disease affects many people worldwide with little known about the intricate mechanisms driving the pathology, making it difficult to develop novel therapies. Improving the understanding of airway smooth muscle and extracellular matrix (ECM) interactions is key to developing treatments for this leading cause of death. With currently no relevant or controllable in vivo or in vitro models to investigate cell-ECM interactions in the small airways, the development of a biomimetic in vitro model with cell attachment, signaling, and organization is needed. The goal of this study was to create a biologically and structurally relevant in vitro model of small airway smooth muscle. In order to achieve this goal, a scaffold was engineered from synthetic poly-l-lactic acid (PLLA) and decellularized pig lung ECM (PLECM). PLECM scaffolds have improved physical characteristics over synthetic scaffolds, by exhibiting a significant decrease in the elastic modulus and an increase in hydrophilicity. Histological staining and SDS-PAGE showed that essential proteins or protein fragments found in natural ECM were present after processing. Human bronchial smooth muscle cells (HBSMCs) seeded onto PLECM 3D scaffolds formed confluent layers and maintained a contractile phenotype, as demonstrated by the organized arrangement of actin filaments within the cell and expected contractile protein expression of calponin 1. HBSMCs cultured on electrospun PLECM scaffold also increased alpha-1 type 1 collagen compared to those cultured on PLLA scaffolds. In summary, this research demonstrates that a PLLA/PLECM composite electrospun mat is a promising tool to produce an in vitro model of the airway with the potential for a better understanding of bronchiole smooth muscle behavior in diseased or normal states.


Comparative Evolution of an Archetypal Adaptive Radiation: Innovation and Opportunity in Anolis Lizards.
Adaptive radiation is a widely recognized pattern of evolution wherein substantial phenotypic change accompanies rapid speciation. Adaptive radiation may be triggered by environmental opportunities resulting from dispersal to new areas or via the evolution of traits, called key innovations, that allow for invasion of new niches. Species sampling is a known source of bias in many comparative analyses, yet classic adaptive radiations have not been studied comparatively with comprehensively sampled phylogenies. In this study, we use unprecedented comprehensive phylogenetic sampling of Anolis lizard species to examine comparative evolution in this well-studied adaptive radiation. We compare adaptive radiation models within Anolis and in the Anolis clade and a potential sister lineage, the Corytophanidae. We find evidence for island (i.e., opportunity) effects and no evidence for trait (i.e., key innovation) effects causing accelerated body size evolution within Anolis. However, island effects are scale dependent: when Anolis and Corytophanidae are analyzed together, no island effect is evident. We find no evidence for an island effect on speciation rate and tenuous evidence for greater speciation rate due to trait effects. These results suggest the need for precision in treatments of classic adaptive radiations such as Anolis and further refinement of the concept of adaptive radiation.


Psychological distress and associated factors among the attendees of traditional healing practices in Jinja and Iganga districts, Eastern Uganda: a cross-sectional study.
Mental health problems are a major public health concern worldwide. Evidence shows that African communities, including Uganda, use both modern and traditional healing systems. There is limited literature about the magnitude of psychological distress and associated factors among attendees of traditional healing practices. This study aimed to determine the prevalence and associated factors of psychological distress among attendees of traditional healing practices in two districts in Uganda. Face-to-face interviews with the Lusoga version of the Self Reporting Questionnaire (SRQ-20) were carried out with 400 patients over the age of 18 years attending traditional healing in Iganga and Jinja districts in Eastern Uganda. Patients were recruited consecutively in all the traditional healers' shrines that could be visited in the area. Persons with 6 or more positive responses to the SRQ were identified as having psychological distress. Prevalence was estimated and odds ratios of having psychological distress were obtained with multiple logistic regression analysis. 387 questionnaire responses were analyzed. The prevalence of psychological distress in connection with attendance at the traditional healers' shrines was 65.1%. Having a co-wife and having more than four children were significantly associated with psyclogical distress. Among the socioeconomic indicators, lack of food and having debts were significantly associated with psychological distress. The distressed group was more likely to need explanations for ill health. Those who visited both the healer and a health unit were less likely to be distressed. This study provides evidence that a substantial proportion of attendees of traditional healing practices suffer from psychological distress. Associated factors include poverty, number of children, polygamy, reason for visiting the healer and use of both traditional healing and biomedical health units. These findings may be useful for policy makers and biomedical health workers for the engagement with traditional healers.


Evaluation of Compass as a comparative mapping tool for ESTs using horse radiation hybrid maps.
Loci for 9322 equine expressed sequence tags (ESTs) were predicted using the Comparative Mapping by Annotation and Sequence Similarity (Compass) strategy in order to evaluate the programme's ability to make accurate locus predictions in species with comparative gene maps. Using human genome sequence information from Build 35 (May 2004) and published marker information from the radiation hybrid (RH) maps for equine chromosomes (ECA) 17 and X, 162 ESTs were predicted to locations on ECA17 and 328 ESTs to locations on ECAX by selection of the 'top blast hit'. The locations of 30 ESTs were assessed experimentally by RH mapping analysis to evaluate the accuracy of the Compass predictions. The data revealed that 53% (16 of 30) of the ESTs predicted on ECA17 and ECAX mapped to those chromosomes. Analysis of the results suggested the need to identify expressed orthologous sequences in order to generate more accurate predictions for ESTs. Locus predictions were reassessed with three modifications to the Compass strategy's orthologue selection parameters. Selection of the 'top gene hit' improved accuracy to 72% (21 of 29), while selection of the 'top expressed gene hit' improved accuracy to 86% (24 of 28). Using the default Compass parameters with the UniGene database improved prediction accuracy to 96% (22 of 23); however, this level of accuracy came with a substantial decrease in the total number of predictions. When used with optimized prediction parameters, the Compass strategy can be a practical in silico map location prediction tool for large EST sample sets from unsequenced animal genomes.


Characterization of impulse propagation at the microscopic level across geometrically defined expansions of excitable tissue: multiple site optical recording of transmembrane voltage (MSORTV) in patterned growth heart cell cultures.
Impulse propagation across sudden expansions of excitable tissue has been shown to exhibit various forms of conduction disturbance on a macroscopic scale, ranging from small delays to unidirectional or complete conduction block. With the present study, we attempted to characterize systematically the dependence of impulse propagation on the geometry of the underlying excitable tissue on a microscopic scale by investigating the spatio-temporal pattern of transmembrane voltage changes associated with impulse propagation from a narrow cell strand to a large cell area using multiple site optical recording of transmembrane voltage (MSORTV) in conjunction with patterned growth of neonatal rat heart cells in culture. While action potential propagation was smooth in the case of funneled expansions, delays of variable size occurred during propagation into rectangular or incised expansions. Close to the abrupt expansion, which functionally represented an increased electrical load to the narrow cell strand, the delays were accompanied by marked distortions of the action potential upstroke, exhibiting, in extreme cases, an initial depolarization to 50% followed by a delayed secondary depolarization to 100% of the full-signal amplitude. These distortions, which were based on bidirectional electrotonic interactions across the transition, were maximal immediately downstream from the expansion. The maximal slowing of impulse conduction across abrupt expansions was, in agreement with recently published results obtained from two-dimensional computer simulations, always situated in the expanded region. At high stimulation rates, the delays sometimes turned into intermittent unidirectional blocks, as revealed by reverse stimulation. These blocks were always characterized by a marked abbreviation of the action potentials upstream from the region causing the block which might, in an appropriate network, facilitate reentry because of the associated shortening of the refractory period. Because the patterns were composed of cells having identical membrane properties, the results show that the local action potential shape can be modulated profoundly by the two-dimensional architecture of the underlying cell ensemble alone.


ARID1A mutation/ARID1A loss is associated with a high immunogenic profile in clear cell ovarian cancer.
ARID1A mutation is frequently found in clear cell ovarian cancer (CCC) and endometrioid ovarian cancer (EC). Anti-PD-1 monotherapy has been found to have limited efficacy in epithelial ovarian cancer; however, anti-PD-1 therapy showed significant clinical benefit in some CCC. We sought to define the relationship of ARID1A mutation/ARID1A expression to the immunogenic profile of different histologic subtypes of ovarian cancer. We performed next-generation sequencing of 160 cancer-related genes. Also, we analyzed the immunohistochemical status of ARID1A, PD-L1, and CD8 with survival in different histologic subtypes of ovarian cancer in a total of 103 cases. ARID1A mutation was found in 0% of the high-grade serous ovarian cancer (HGSC) (n = 36), 41.5% of the CCC (n = 41), 45.0% of the EC (n = 20), and 33.3% of the mucinous ovarian cancer (MC) (n = 6) cases. ARID1A loss was found in 19.4% of the HGSC, 75.6% of the CCC, 60.0% of the EC and 0% of the MC cases. ARID1A mutation was found to be associated with high PD-L1 (p < 0.001) or CD8 levels (p < 0.001) in CCC but not in other histologic subtypes. Meanwhile, ARID1A loss was associated with high PD-L1 or CD8 levels in CCC (p < 0.001) and HGSC (p < 0.001) but not in EC and MC. In addition, ARID1A mutation was associated with high tumor mutation burden in CCC (p = 0.006). ARID1A mutation/ARID1A expression is associated with immune microenvironmental factors in CCC but not in EC. ARID1A status can be a biomarker for selecting candidates for immune checkpoint blockade in CCC.


Sheep prions with molecular properties intermediate between classical scrapie, BSE and CH1641-scrapie.
Efforts to differentiate bovine spongiform encephalopathy (BSE) from scrapie in prion infected sheep have resulted in effective methods to decide about the absence of BSE. In rare instances uncertainties remain due to assumptions that BSE, classical scrapie and CH1641-a rare scrapie variant-could occur as mixtures. In field samples including those from fallen stock, triplex Western blotting analyses of variations in the molecular properties of the proteinase K resistant part of the disease‑associated form of prion protein (PrP(res)) represents a powerful tool for quick discrimination purposes. In this study we examined 7 deviant ovine field cases of scrapie for some typical molecular aspects of PrP(res) found in CH1641‑scrapie, classical scrapie and BSE. One case was most close to scrapie with respect to molecular mass of its non-glycosylated fraction and N-terminally located 12B2‑epitope content. Two cases were unlike classical scrapie but too weak to differentiate between BSE or CH1641. The other 4 cases appeared intermediate between scrapie and CH1641 with a reduced molecular mass and 12B2‑epitope content, together with the characteristic presence of a second PrP(res) population. The existence of these 2 PrP(res) populations was further confirmed through deglycosylation by PNGaseF. The findings indicate that discriminatory diagnosis between classical scrapie, CH1641 and BSE can remain inconclusive with current biochemical methods. Whether such intermediate cases represent mixtures of TSE strains should be further investigated e.g. in bioassays with rodent lines that are varying in their susceptibility or other techniques suitable for strain typing.


Tumor Regression and Cure Depends on Sustained Th1 Responses.
While immunomodulatory monoclonal antibodies (mAbs) have therapeutic efficacy against many tumors, few patients are cured. Attempting to improve their therapeutic efficacy we have applied the TC1 mouse lung carcinoma model and injected established subcutaneous tumors intratumorally with 3 weekly doses of various combinations of mAbs. Combinations of mAbs to CTLA4/PD1/CD137 (the 3 mAb combination) and to CTLA4/PD1/CD137/CD19 (the 4 mAb combination) were most efficacious to induce complete regression of both the injected tumor and an untreated tumor in the same mouse. Tumor cure was consistently associated with shifting a Th2 to a Th1 response in tumor-draining lymph nodes and spleen and it involved epitope specific and long-lived memory T cells as well as M1 macrophages. This shift and accompanying tumor rejection was harder to achieve as the treated tumors increased in size. Relapse of tumors which had initially regressed following treatment with immunomodulatory mAbs was associated with return of a Th2 microenvironment in tumors, tumor-draining lymph nodes and spleens rather than the emergence of immune-resistant tumor cells. While mAbs to CTLA4 plus PD-1 were therapeutically ineffective, combining the 2 of them with intraperitoneal cisplatin, 10 mg/kg, induced long-term complete tumor regression in most mice with small TC1 tumors and the therapeutic efficacy against larger tumors improved by administrating cisplatin together with the 3 or 4 mAb combination.


Pulmonary barotrauma in SCUBA diving-related fatalities: a histological and histomorphometric analysis.
Arterial gas embolism following pulmonary barotrauma occurs in 13-24% of cases of diving deaths. The study aimed to evaluate the usefulness of a histomorphometric digital analysis in the detection of air space over-distension due to pulmonary barotrauma. The study was performed on lung parenchyma specimens of 12 divers: six had died due to arterial gas embolism following pulmonary barotrauma (mean age at death of 54 years, range of 41-61 years), and six had drowned in saltwater without a diagnosis of pulmonary barotrauma (mean age at death of 54 years, range of 41-66 years) (positive controls). For negative controls, six cases of non-SCUBA divers (mean age of death of 42 years, range of 23-55 years) who died of intracerebral haemorrhage were evaluated. No significant differences were observed in the characteristics of the air spaces between control groups (positive and negative). However, differences were observed in the area occupied by air spaces and the percentage of air space area when we compared the case group to the controls (p < 0.01); and there was a slight difference in the maximum and minimum diameters of air space (p < 0.05). The mean area occupied by air spaces and the mean percentage of air space were the most useful for discriminating pulmonary barotrauma from other causes of death (100% sensitivity and 91.7% specificity). Based on our study, inclusion of an increased pattern of air spaces as a possible diagnostic criterion for pulmonary barotrauma would be useful in discerning the cause of diving death.


Effect of viral storm in patients admitted to intensive care units with severe COVID-19 in Spain: a multicentre, prospective, cohort study.
The contribution of the virus to the pathogenesis of severe COVID-19 is still unclear. We aimed to evaluate associations between viral RNA load in plasma and host response, complications, and deaths in critically ill patients with COVID-19. We did a prospective cohort study across 23 hospitals in Spain. We included patients aged 18 years or older with laboratory-confirmed SARS-CoV-2 infection who were admitted to an intensive care unit between March 16, 2020, and Feb 27, 2021. RNA of the SARS-CoV-2 nucleocapsid region 1 (N1) was quantified in plasma samples collected from patients in the first 48 h following admission, using digital PCR. Patients were grouped on the basis of N1 quantity: VIR-N1-Zero (<1 N1 copies per mL), VIR-N1-Low (1-2747 N1 copies per mL), and VIR-N1-Storm (>2747 N1 copies per mL). The primary outcome was all-cause death within 90 days after admission. We evaluated odds ratios (ORs) for the primary outcome between groups using a logistic regression analysis. 1068 patients met the inclusion criteria, of whom 117 had insufficient plasma samples and 115 had key information missing. 836 patients were included in the analysis, of whom 403 (48%) were in the VIR-N1-Low group, 283 (34%) were in the VIR-N1-Storm group, and 150 (18%) were in the VIR-N1-Zero group. Overall, patients in the VIR-N1-Storm group had the most severe disease: 266 (94%) of 283 patients received invasive mechanical ventilation (IMV), 116 (41%) developed acute kidney injury, 180 (65%) had secondary infections, and 148 (52%) died within 90 days. Patients in the VIR-N1-Zero group had the least severe disease: 81 (54%) of 150 received IMV, 34 (23%) developed acute kidney injury, 47 (32%) had secondary infections, and 26 (17%) died within 90 days (OR for death 0·30, 95% CI 0·16-0·55; p<0·0001, compared with the VIR-N1-Storm group). 106 (26%) of 403 patients in the VIR-N1-Low group died within 90 days (OR for death 0·39, 95% CI 0·26-0·57; p<0·0001, compared with the VIR-N1-Storm group). The presence of a so-called viral storm is associated with increased all-cause death in patients admitted to the intensive care unit with severe COVID-19. Preventing this viral storm could help to reduce poor outcomes. Viral storm could be an enrichment marker for treatment with antivirals or purification devices to remove viral components from the blood. Instituto de Salud Carlos III, Canadian Institutes of Health Research, Li Ka-Shing Foundation, Research Nova Scotia, and European Society of Clinical Microbiology and Infectious Diseases. For the Spanish translation of the abstract see Supplementary Materials section.


Simulation of Airflow in an Idealized Emphysematous Human Acinus.
Emphysema is the permanent enlargement of air spaces in the respiratory regions of the lung due to destruction of the inter-alveolar septa. The progressive coalescence of alveoli and alveolar ducts into larger airspaces leads to the disruption of normal airway wall motion and airflow rates within the pulmonary acinus. To contribute to the understanding of the individual effects of emphysema during its earliest stages, computational fluid dynamics (CFD) simulations of airflow in mathematically derived models of the pulmonary acinus were performed. The here generated computational domain consists of two generations of alveolar ducts within the pulmonary acinus, with alveolar geometries approximated as closely packed, 14-sided polygons. Physiologically realistic airflow rates and wall motions were used to study airflow patterns within subsequent generations of alveolar ducts during the inspiratory and expiratory phases of the breathing cycle. The effects of progressive emphysema on the airway wall motion and flow rates were simulated by sequentially removing all alveolar septa within each alveolar duct. Parametric studies were presented to independently assess the relative influence of progressive septal destruction of airway motion and flow rates. The results illustrate that septal destruction lowers the flow resistance through the alveolar ducts but has little influence on the mass transport of oxygen into the alveoli. Septal destruction has a net effect on the flow field by favoring the development of recirculatory flow patterns in individual alveoli.


Brain metastases in endometrial carcinoma.
Brain metastases of endometrial carcinoma are rare. They tend to follow an unpredictable dissemination pattern, due to the different pathways that tumor cells can follow and to certain factors that influence their spread. When brain metastases occur, usually the disease is widely disseminated, but there are some exceptions such as the case presented here. We report the case of a 76-year-old woman who presented a single occipital metastasis 18 months after a hysterectomy because of an endometrial carcinoma. No other metastases were found. We discuss the different dissemination pathways of endometrial carcinoma. Some hypotheses try to explain what determines the site of preference of distant metastases. The most accepted hypothesis suggests the presence of specific tumor cell receptors and endothelium receptors on the target organs, which would explain the preference of different cancers for different targets and for different areas inside the same target organ. Although brain metastases of endometrial carcinoma are rare and they usually occur in widely disseminated disease, they should be considered in patients with a single brain mass, even when the primary disease is unknown.


Adaptive classification for brain computer interfaces.
In this paper we evaluate the performance of a new adaptive classifier for the use within a Brain Computer-Interface (BCI). The classifier can either be adaptive in a completely unsupervised manner or using unsupervised adaptation in conjunction with a neuronal evaluation signal to improve adaptation. The first variant, termed Adaptive Linear Discriminant Analysis (ALDA), updates mean values as well as covariances of the class distributions continuously in time. In simulated as well as experimental data ALDA substantially outperforms the non-adaptive LDA. The second variant, termed Adaptive Linear Discriminant Analysis with Error Correction (ALDEC), extends the unsupervised algorithm with an additional independent neuronal evaluation signal. Such a signal could be an error related potential which indicates when the decoder did not classify correctly. When the mean values of the class distributions circle around each other or even cross their way, ALDEC can yield a substantially better adaptation than ALDA depending on the reliability of the error signal. Given the non-stationarity of EEG signals during BCI control our approach might strongly improve the precision and the time needed to gain accurate control in future BCI applications.


myo-Inositol homeostasis in the human fetus.
The concentration of myo-inositol in the serum of pregnant women (21.4 microM) remained unchanged throughout pregnancy and was not significantly different from that in the serum of nonpregnant women (24.5 microM). myo-Inositol concentrations in mixed cord serum averaged 125 microM at midgestation. At term, the concentration of myo-inositol was 60.2 microM in the umbilical artery and 45.4 microM in the umbilical vein. In every umbilical cord examined, the ratio of myo-inositol concentration in umbilical artery to that in umbilical vein exceeded 1.0. Activity of a putative regulatory enzyme in the biosynthesis of myo-inositol from glucose, ie, glucose 6-P:inositol 1-P cyclase (cyclase), was measured in placenta, fetal lung, and fetal liver. In placenta at midgestation and at term, cyclase activity was 87.8 and 90.7 nmol X g-1 tissue X h-1, respectively. Cyclase activity in fetal lung and liver at midgestation was 54.6 and 54.4 nmol X g-1 tissue X h-1, respectively. The gestational decline in the concentration of myo-inositol in fetal serum may represent a decreasing availability of myo-inositol to the fetal lungs and could be important in the mechanism whereby the amounts of phosphatidylglycerol and phosphatidylinositol in lung surfactant are regulated during fetal lung development.


In situ construction of hierarchical Co/MnO@graphite carbon composites for highly supercapacitive and OER electrocatalytic performances.
The development of new electrode materials with high specific capacity for excellent supercapacitive storage and energy conversion is highly desirable. The combination of metal and metal oxide with carbon is an effective strategy to achieve active bimetallic nanocatalysts. Herein, we developed a facile method to synthesize CoxMn1-xO@GC and Co/MnO@GC nanocomposites by an in situ conversion of Co-Mn PBAs. The as-prepared carbon hybrids, especially the resulting Co/MnO@GC carbonized under 700 °C (Co/MnO@GC-700), preserve the nanocubic morphology of Co-Mn PBAs and show excellent supercapacitance and OER performance. Specifically, an outstanding specific capacitance of 2275 F g-1 can be obtained with Co/MnO@GC-700 as the electrode material at a current density of 4 A g-1. When used as OER catalysts, Co/MnO@GC-700 shows a low overpotential of only 358 mV at 10 mA cm-2 in 1 M KOH. Moreover, a fabricated asymmetric supercapacitor device (ASC device), in combination with active carbon, shows a high cell voltage of 1.7 V and a considerably high specific capacitance of 246 F g-1 at 2 A g-1. Our nanoarchitecture design derived from PBAs provides a new opportunity for future applications in high-performance energy storage and transformation systems.


Chlamydia pneumoniae augments the oxidized low-density lipoprotein-induced death of mouse macrophages by a caspase-independent pathway.
Chlamydia pneumoniae is a common respiratory pathogen that is associated with an increased risk of cardiovascular disease. However, the mechanisms by which C. pneumoniae contributes to cardiovascular disease have not been determined yet. C. pneumoniae infection may accelerate the death of cells within atherosclerotic lesions and contribute to the formation of unstable lesions. To test this hypothesis, the impact of C. pneumoniae infection on the death of lipid-loaded mouse macrophages was investigated. It was observed that RAW 264.7 cells are highly susceptible to the toxic effects of oxidized low-density lipoprotein (LDL) and exhibit markers of cell death within 24 h of treatment with as little as 5 microg/ml oxidized LDL. Subsequent infection with either live C. pneumoniae or heat-killed or UV-inactivated C. pneumoniae at a low multiplicity of infection for 24 to 72 h stimulated both additional binding of annexin V and the uptake of propidium iodide. Thus, C. pneumoniae augments the effects of oxidized LDL on cell death independent of a sustained infection. However, unlike oxidized LDL, C. pneumoniae infection does not activate caspase 3 or induce formation of the mitochondrial transition pore or the fragmentation of DNA, all of which are classical markers of apoptosis. Furthermore, primary bone marrow macrophages isolated from mice deficient in Toll-like receptor 2 (TLR-2) but not TLR-4 are resistant to C. pneumoniae-induced death. These data suggest that C. pneumoniae kills cells by a caspase-independent pathway and that the process is potentially mediated by activation of TLR-2.


Highly birefringent elliptical-hole photonic crystal fiber with squeezed hexagonal lattice.
We propose a novel polarization-maintaining index-guiding photonic crystal fiber (PCF). It is composed of a solid silica core and a cladding with squeezed-hexagonal-lattice elliptical air holes. Using a full-vector finite-element method, we study the modal birefringence of the fundamental modes in such PCFs. Numerical result shows that very high modal birefringence with a magnitude of the order of 10(-2) around 1550 nm has been obtained. Furthermore, large normal dispersion appears over a wide range of wavelengths in both orthogonal polarizations.


Course and characteristics of anaemia in patients with rheumatoid arthritis of recent onset.
To describe the incidence, cause, and course of anaemia in rheumatoid arthritis (RA). Medical records of 225 patients who received a diagnosis of RA between 1990 and 1992 were reviewed longitudinally for mention of anaemia. Anaemia was classified as anaemia of chronic disease if ferritin concentrations reflected adequate body iron stores. Among iron depleted anaemic patients, iron deficiency anaemia was identified using the response to iron supplementation. Anaemia developed in 64% of the patients, mostly within 18 months of follow up, but disappeared again in 54% of those patients. The prevalence of anaemia varied from 39% to 53% throughout follow up. Iron depletion was found in 38% of anaemic patients; 40% of them did not recover from their anaemia after iron supplementation and were classified as having anaemia of chronic disease. Anaemia of chronic disease thus caused 77% and iron deficiency anaemia 23% of observed anaemia. Recovery from anaemia occurred in 42% of the patients with anaemia of chronic disease and in 72% of iron depleted patients after iron supplementation. Anaemic patients, particularly those with anaemia of chronic disease, had a significantly greater number of the American College of Rheumatism criteria for RA, significantly more erosive joint damage, and significantly increased concentrations of serum rheumatoid factor than patients without anaemia. Anaemia appeared as a frequent and dynamic manifestation. Recovery and recurrence of anaemia was observed throughout follow up, leading to a longstanding and relatively high prevalence of the condition. Iron deficiency was diagnosed frequently and follow up revealed a considerable overlap with anaemia of chronic disease, making this the most important cause of anaemia in RA. Recovery from anaemia occurred more frequently in iron depleted anaemic patients than in those with anaemia of chronic disease. Anaemic patients, particularly those with anaemia of chronic disease, seemed to have a more serious course of their RA compared with non-anaemic patients.


Psychological Resilience, Mental Health, and Inhibitory Control Among Youth and Young Adults Under Stress.
Psychological resilience allows one to cope successfully with adversities occurring during stressful periods, which may otherwise trigger mental illness. Recent models suggest that inhibitory control (IC), the executive control function which supports our goal-directed behavior and regulates our emotional response, may underlie resilience. However, the ways in which this is manifested during stressful situations in real life is still unclear. Here, we examined the relationship between IC, psychological resilience, psychological distress, and anxiety among 138 female and male participants in a stressful situation: during their initial combat training in the military. Using a mobile app, we assessed IC using emotional and non-emotional variations of the Go/No-Go task. Psychological resilience, psychological distress, and anxiety were assessed using mobile versions of self-report questionnaires. We found that psychological resilience is significantly correlated with non-emotional IC (r = 0.24, p < 0.005), but not with emotional IC; whereas, psychological distress and anxiety are correlated with emotional IC (r = -0.253, p < 0.005 and r = -0.224, p < 0.01, for psychological distress and anxiety, respectively), but not with non-emotional IC. A regression model predicting emotional IC confirmed non-emotional IC and distress as unique contributors to the variance, but not psychological distress. In addition, associations between psychological distress and emotional IC were found only for female participants. Collectively, the results clarify the link between IC, resilience, and mental health in real-life stressful situations, showing separate mechanisms of IC involved in resilience on the one hand, and mental health on the other hand. These results have implications for building mobile resilience interventions for youth and young adults facing stressful situations.


Nanoscopic Terraces, Mesas, and Ridges in Freely Standing Thin Films Sculpted by Supramolecular Oscillatory Surface Forces.
Freely standing thin liquid films containing supramolecular structures including micelles, nanoparticles, polyelectrolyte-surfactant complexes, and smectic liquid crystals undergo drainage via stratification. The layer-by-layer removal of these supramolecular structures manifests as stepwise thinning over time and a coexistence of domains and nanostructures of discretely different thickness. The layering of supramolecular structures in confined thin films contributes additional non-DLVO, supramolecular oscillatory surface forces to disjoining pressure, thus influencing both drainage kinetics and stability of thin films. Understanding and characterizing the spontaneous creation and evolution of nanoscopic topography of stratifying, freely standing thin liquid films have been long-standing challenges due to the absence of experimental techniques with the requisite spatial (thickness <10 nm) and temporal resolution (<1 ms). Using Interferometry Digital Imaging Optical Microscopy (IDIOM) protocols developed herein, we visualize and characterize size, shape, and evolution kinetics of nanoscopic mesas, terraces, and ridges. The exquisite thickness maps created using IDIOM protocols provide much needed and unprecedented insights into the role of supramolecular oscillatory surface forces in driving growth of such nanostructures as well as in controlling properties and stability of freely standing thin films and, more generally, of colloidal dispersions like foams.


The effects of pole walking on health in adults: a systematic review.
The aim of this systematic review was to summarize the effects of pole walking (PW) programs on physical and psychosocial health. Randomized controlled and controlled trials were identified from literature searches in PubMed, Cochrane library, EMBASE, SPORTdiscuss, CINAHL and PEDRO. A total of 14 articles from 13 studies met the inclusion criteria. Eleven of the included studies had a quality score of 50% or higher. Most studies included mid to older aged men and women in clinical populations with various medical conditions. Only two studies included nonclinical populations. The majority of the PW programs consisted of supervised group sessions performed two to three times weekly for 8 weeks or longer. Most studies investigated the effects of PW on both physical and psychosocial health and the majority examined effects on four to five outcomes. The effects of PW on cardiorespiratory fitness were most extensively studied. The most frequently examined psychosocial measure was quality of life. All studies reported at least one beneficial effect of PW compared with the control group. The results of this systematic review indicate that PW programs have some beneficial effects on both physical and psychosocial health in adults with and without clinical conditions.


Major dietary patterns and the risk of colorectal cancer in women.
Several foods and nutrients have been implicated in the development of colon and rectal cancers. In this study, we prospectively assessed the associations between major dietary patterns and the risks of these 2 cancers in women. Using dietary information collected in 1984, 1986, 1990, and 1994 from 76 402 women aged 38 to 63 years without a history of cancer in 1984, we conducted factor analysis and identified 2 major dietary patterns: "prudent" and "Western." We calculated factor scores for each participant and examined prospectively the associations between dietary patterns and colon and rectal cancer risks. The prudent pattern was characterized by higher intakes of fruits, vegetables, legumes, fish, poultry, and whole grains, while the Western pattern, by higher intakes of red and processed meats, sweets and desserts, french fries, and refined grains. During 12 years of follow-up, we identified 445 cases of colon cancer and 101 cases of rectal cancer. After adjusting for potential confounders, we observed a relative risk for colon cancer of 1.46 (95% confidence interval, 0.97-2.19) when comparing the highest with the lowest quintiles of the Western pattern (P value for trend across quintiles, .02). The prudent pattern had a nonsignificant inverse association with colon cancer (relative risk for fifth quintile compared with the first, 0.71; 95% confidence interval, 0.50-1.00; P for trend across quintiles, .31). We did not observe any significant association between dietary patterns and rectal cancer. We found a significant positive association between the Western dietary pattern and the risk of colon cancer.


AMPA receptor upregulation in the nucleus accumbens shell of cocaine-sensitized rats depends upon S-nitrosylation of stargazin.
Behavioral sensitization to cocaine is associated with increased AMPA receptor (AMPAR) surface expression in the nucleus accumbens (NAc). This upregulation is withdrawal-dependent, as it is not detected on withdrawal day (WD) 1, but is observed on WD7-21. Its underlying mechanisms have not been clearly established. Nitric oxide (NO) regulates AMPAR trafficking in the brain by S-nitrosylation of the AMPAR auxiliary subunit, stargazin, leading to increased AMPAR surface expression. Our goal was to determine if stargazin S-nitrosylation contributes to AMPAR upregulation during sensitization. First, we measured stargazin S-nitrosylation in NAc core and shell subregions on WD14 after 8 daily injections of saline or 15 mg/kg cocaine. Stargazin S-nitrosylation was markedly increased in NAc shell but not core. To determine if this is associated with AMPAR upregulation, rats received 8 cocaine or saline injections followed by twice-daily treatments with vehicle or the nitric oxide synthase inhibitor l-NAME (50 mg/kg) on WD1-6, the time when AMPAR upregulation is developing in cocaine-exposed rats. Cocaine/vehicle rats showed elevated stargazin and GluA1 surface expression on WD7 compared to saline/vehicle rats; the GluA1 increase was more robust in core, while stargazin increased more robustly in shell. These effects of cocaine were attenuated in shell but not core when cocaine injections were followed by l-NAME treatment on WD1-6. Together, these results indicate that elevated S-nitrosylation of stargazin contributes to AMPAR upregulation during sensitization selectively in the NAc shell. It is possible that AMPAR upregulation in core involves a different TARP, γ4, which also upregulates in the NAc of sensitized rats.


Propionate activates multiple ion transport mechanisms in the HT29-18-C1 human colon cell line.
Short-chain fatty acids (SCFAs) are the major solutes and the major anions in the colonic lumen. We studied the response of suspended HT29-18-C1 cells (an epithelial cell line derived from a human colon carcinoma) to SCFA exposure. Cellular response was evaluated by measurement of cell volume (Coulter counter), intracellular pH [pHi; measured fluorometrically with 2',7'-bis(2-carboxyethyl)-5-(6)-carboxyfluorescein (BCECF)], and intracellular Na+, K+, and Cl- content (flame photometry and chloride titrator). Exposure to 130 mM propionate in isosmotic medium causes a rapid decrease in pHi and activates pHi recovery via amiloride-sensitive Na-H exchange. In the presence of propionate, Na-H exchange also causes cell swelling to a peak volume 11% above control cells and causes a 2.8-fold increase in intracellular Na+ content. After peak swelling, a regulatory-volume decrease (RVD) significantly reduced volume and intracellular Na+ returned to baseline. Other SCFAs (acetate, butyrate, and valerate) also elicit swelling and RVD. Activation of the Na(+)-K(+)-adenosinetriphosphatase (ATPase) is required to return Na+ to normal levels and to indirectly provide ion gradients required for propionate-induced RVD, but Na(+)-K(+)-ATPase activity does not directly mediate RVD. When 1 mM 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) is added in the presence of propionate, RVD was inhibited and cell Na+ content increased. Cl- depletion inhibited propionate-induced RVD and diminished the effect of SITS.


Farnesoid X receptor-induced lysine-specific histone demethylase reduces hepatic bile acid levels and protects the liver against bile acid toxicity.
Bile acids (BAs) function as endocrine signaling molecules that activate multiple nuclear and membrane receptor signaling pathways to control fed-state metabolism. Since the detergent-like property of BAs causes liver damage at high concentrations, hepatic BA levels must be tightly regulated. Bile acid homeostasis is regulated largely at the level of transcription by nuclear receptors, particularly the primary BA receptor, farnesoid X receptor, and small heterodimer partner, which inhibits BA synthesis by recruiting repressive histone-modifying enzymes. Although histone modifiers have been shown to regulate BA-responsive genes, their in vivo functions remain unclear. Here, we show that lysine-specific histone demethylase1 (LSD1) is directly induced by BA-activated farnesoid X receptor, is recruited to the BA synthetic genes Cyp7a1 and Cyp8b1 and the BA uptake transporter gene Ntcp, and removes a gene-activation marker, trimethylated histone H3 lysine-4, leading to gene repression. Recruitment of LSD1 was dependent on small heterodimer partner, and LSD1-mediated demethylation of trimethylated histone H3 lysine-4 was required for additional repressive histone modifications, acetylated histone 3 on lysine 9 and 14 deacetylation, and acetylated histone 3 on lysine 9 methylation. A BA overload, feeding 0.5% cholic acid chow for 6 days, resulted in adaptive responses of altered expression of hepatic genes involved in BA synthesis, transport, and detoxification/conjugation. In contrast, adenovirus-mediated downregulation of hepatic LSD1 blunted these responses, which led to substantial increases in liver and serum BA levels, serum alanine aminotransferase and aspartate aminotransferase levels, and hepatic inflammation. This study identifies LSD1 as a novel histone-modifying enzyme in the orchestrated regulation mediated by the farnesoid X receptor and small heterodimer partner that reduces hepatic BA levels and protects the liver against BA toxicity.


Soft electrochemical bubble actuator with liquid metal electrode using an embodied hydrogel pneumatic source.
Soft pneumatic actuators-such as those used for soft robotics-achieve actuation by inflation of pneumatic chambers. Here, we report the use of the electrochemical reduction of water to generate gaseous products that inflate pneumatic chambers. Whereas conventional pneumatic actuators typically utilize bulky mechanical pumps, the approach here utilizes only electricity. In contrast to dielectric actuators, which require ∼kV to actuate, the electrochemical approach uses a potential of a few volts. The applied potential converts liquid water-a safe, abundant, and cheap fuel-into hydrogen gas. Since the chambers are constructed of hydrogel, the body of the actuator provides an abundant supply of water that ultimately converts to gas. The use of liquid metal for the electrode makes the entire device soft and ensures intimate contact between the chamber walls and the electrode during inflation. The device can inflate in tens of seconds, which is slower than other pneumatic approaches, but much faster than actuating hydrogels via principles of swelling. The actuation volume can be predicted and controlled based on the input parameters such as time and voltage. The actuation shape and position can also be controlled by the position of the electrodes and the geometry of the device. Such actuators have the potential to make tether-less (pump-free), electrically-controlled soft devices that can even operate underwater.