Inverse association of plasma IL-13 and inflammatory chemokines with lung function impairment in stable COPD: a cross-sectional cohort study.
Chronic obstructive pulmonary disease (COPD) is a heterogeneous syndrome characterized by varying degrees of airflow limitation and diffusion impairment. There is increasing evidence to suggest that COPD is also characterized by systemic inflammation. The primary goal of this study was to identify soluble proteins in plasma that associate with the severity of airflow limitation in a COPD cohort with stable disease. A secondary goal was to assess whether unique markers associate with diffusion impairment, based on diffusion capacity of carbon monoxide (DLCO), independent of the forced expiratory volume in 1 second (FEV1). A cross sectional study of 73 COPD subjects was performed in order to examine the association of 25 different plasma proteins with the severity of lung function impairment, as defined by the baseline measurements of the % predicted FEV1 and the % predicted DLCO. Plasma protein concentrations were assayed using multiplexed immunobead-based cytokine profiling. Associations between lung function and protein concentrations were adjusted for age, gender, pack years smoking history, current smoking, inhaled corticosteroid use, systemic corticosteroid use and statin use. Plasma concentrations of CCL2/monocyte chemoattractant protein-1 (CCL2/MCP-1), CCL4/macrophage inflammatory protein-1 beta (CCL4/MIP-1 beta), CCL11/eotaxin, and interleukin-13 (IL-13) were inversely associated with the % FEV1. Plasma concentrations of soluble Fas were associated with the % DLCO, whereas CXCL9/monokine induced by interferon-gamma (CXCL9/Mig), granulocyte- colony stimulating factor (G-CSF) and IL-13 showed inverse relationships with the % DLCO. Systemic inflammation in a COPD cohort is characterized by cytokines implicated in inflammatory cell recruitment and airway remodeling. Plasma concentrations of IL-13 and chemoattractants for monocytes, T lymphocytes, and eosinophils show associations with increasing severity of disease. Soluble Fas, G-CSF and CXCL9/Mig may be unique markers that associate with disease characterized by disproportionate abnormalities in DLCO independent of the FEV1.


Characterization of Komagataeibacter Isolate Reveals New Prospects in Waste Stream Valorization for Bacterial Cellulose Production.
Komagataeibacter spp. has been used for the bioconversion of industrial wastes and lignocellulosic hydrolysates to bacterial cellulose (BC). Recently, studies have demonstrated the capacity of Komagataeibacter spp. in the biotransformation of inhibitors found in lignocellulosic hydrolysates, aromatic lignin-derived monomers (LDMs) and acetate. In general, detoxification and BC synthesis from lignocellulosic inhibitors requires a carbon flow from acetyl-coA towards tricarboxylic acid and gluconeogenesis, respectively. However, the related molecular aspects have not yet been identified in Komagataeibacter spp. In this study, we isolated a cellulose-producing bacterium capable of synthesizing BC in a minimal medium containing crude glycerol, a by-product from the biodiesel production process. The isolate, affiliated to Komagataeibacter genus, synthesized cellulose in a minimal medium containing glucose (3.3 ± 0.3 g/L), pure glycerol (2.2 ± 0.1 g/L) and crude glycerol (2.1 ± 0.1 g/L). Genome assembly and annotation identified four copies of bacterial cellulose synthase operon and genes for redirecting the carbon from the central metabolic pathway to gluconeogenesis. According to the genome annotations, a BC production route from acetyl-CoA, a central metabolic intermediate, was hypothesized and was validated using acetate. We identified that when K. rhaeticus ENS9b was grown in a minimal medium supplemented with acetate, BC production was not observed. However, in the presence of readily utilizable substrates, such as spent yeast hydrolysate, acetate supplementation improved BC synthesis.


Gut microbiome, enteric infections and child growth across a rural-urban gradient: protocol for the ECoMiD prospective cohort study.
The functional consequences of the bacterial gut microbiome for child health are not well understood. Characteristics of the early child gut microbiome may influence the course of enteric infections, and enteric infections may change the composition of the gut microbiome, all of which may have long-term implications for child growth and development. We are conducting a community-based birth cohort study to examine interactions between gut microbiome conditions and enteric infections, and how environmental conditions affect the development of the gut microbiome. We will follow 360 newborns from 3 sites along a rural-urban gradient in northern coastal Ecuador, characterising enteric infections and gut microbial communities in the children every 3 to 6 months over their first 2 years of life. We will use longitudinal regression models to assess the correlation between environmental conditions and gut microbiome diversity and presence of specific taxa, controlling for factors that are known to be associated with the gut microbiome, such as diet. From 6 to 12 months of age, we will collect weekly stool samples to compare microbiome conditions in diarrhoea stools versus stools from healthy children prior to, during and after acute enteric infections, using principal-coordinate analysis and other multivariate statistical methods. Ethics approvals have been obtained from Emory University and the Universidad San Francisco de Quito institutional review boards. The findings will be disseminated through conference presentations and peer-reviewed journals.


Toxicity of silver nanoparticles in mouse bone marrow-derived dendritic cells: Implications for phenotype.
Silver nanoparticles (AgNP) are one of the most studied nanoparticles due to their anti-bacterial, -fungal, -viral, -parasitic, and -inflammatory properties. This raises the need to evaluate the toxicity and biological effects of AgNP in the immune system in order to develop new safer biomedical products. In this study, an AgNP formulation currently approved for veterinary applications was applied to mouse bone marrow-derived dendritic cells (BMDC), considered important antigen-presenting cells of the immune system, to evaluate cytotoxicity, genotoxicity, and any significant influence on expression of cellular markers associated with BMDC phenotype and maturation status. The results showed that after 12 h of AgNP exposure, a significant decrease in BMDC viability occurred at the highest concentration tested (1.0 µg AgNP/ml) and at lower doses, the cells maintained membrane integrity and metabolic activity. DNA damage was not significant with any AgNP level aside from the 1.0 µg AgNP/ml level. Regarding phenotype, no differences in expression of CD40 (co-stimulatory molecule highly present in mature BMDC) or in CD273 (a marker for inhibitory T-cell response) were observed. The current results showed that the toxicity of this AgNP formulation was dose-related. The findings also suggest BMDC could maintain structural conservation of co-stimulatory/co-inhibitory surface molecules after 12 h of exposure to this AgNP. This work represents the first step in identifying the toxic effects of this AgNP formulation on dendritic cells.


[The growth of preneoplastic hepatocytes in the spleen of recipient mice].
Hepatocytes and the fraction of non parenchymal cells enriched with oval cells were extracted from the preneoplastic mouse liver at the stage of hyperplastic node formation and implanted into the spleen. In 14-16 months after the transplantation, multiple islets of hepatocytes which replaced up to 25% of the spleen cut area, were found in 57% (4 of 7) and 22% (8 of 36) of recipients respectively. The hepatocytes formed 2-3-cell bulks or solid masses organized into multicellular trabecules, and expressed biliary capillary antigen, albumin and transferrin. In the inoculation of nonparenchymal cell fraction, the growth of hepatic tissue in the spleen depended on the magnitude of hepatocyte admixture to be undetectable in absence of hepatocytes in the donor suspension. The growth of hepatic tissue in spleen was observed following the injection of a small number (3 x 10(-4)-6 x 10(-5)) of live hepatocytes. This fact evidences an extremely high clonogenic potency of clonogenic potency of preneoplastic hepatocytes.


Chromophore-assisted light inactivation of pKi-67 leads to inhibition of ribosomal RNA synthesis.
Expression of the nuclear Ki-67 protein (pKi-67) is strongly associated with cell proliferation. For this reason, antibodies against this protein are widely used as prognostic tools for the assessment of cell proliferation in biopsies from cancer patients. Despite this broad application in histopathology, functional evidence for the physiological role of pKi-67 is still missing. Recently, we proposed a function of pKi-67 in the early steps of ribosomal RNA (rRNA) synthesis. Here, we have examined the involvement of pKi-67 in this process by photochemical inhibition using chromophore-assisted light inactivation (CALI). Anti-pKi-67 antibodies were labelled with the fluorochrome fluorescein 5(6)-isothiocyanate and were irradiated after binding to their target protein. Performing CALI in vitro on cell lysates led to specific cross-linking of pKi-67. Moreover, the upstream binding factor (UBF) necessary for rRNA transcription was also partly subjected to cross-link formation, indicating a close spatial proximity of UBF and pKi-67. CALI in living cells, using micro-injected antibody, caused a striking relocalization of UBF from foci within the nucleoli to spots located at the nucleolar rim or within the nucleoplasm. pKi-67-CALI resulted in dramatic inhibition of RNA polymerase I-dependent nucleolar rRNA synthesis, whereas RNA polymerase II-dependent nucleoplasmic RNA synthesis remained almost unaltered. Our data presented here argue for a crucial role of pKi-67 in RNA polymerase I-dependent nucleolar rRNA synthesis.


[Structure, function and the role of factor XIII in human pathology].
Modern views on the functioning of coagulation system are constantly replenished with the facts obtained using the newest methods of molecular biology and medicine. The purpose of many components of this system is non-uniform and covers the whole spectrum of biochemical reactions responsible for homeostasis. Current data on one of such components--the factor XIII (fibrin stabilizing factor) are stated. Except for the description of the enzyme structure and functions, information is presented about the methods of its detection, which development has resulted in accumulation of knowledge on the role of fibrin stabilizing factor in bleedings and thrombosis genesis. Data are given on hereditary and acquired deficiency of the factor XIII, as well as on conditions accompanied with the increase of its activity.


Prolactin binding in ovariectomy-responsive and ovariectomy-nonresponsive rat mammary carcinoma.
Growth of the transplantable mammary tumor, MTW9, in W/Fu rats is greatly enhanced by elevated serum prolactin concentrations. This report compares the prolactin binding to tumor membranes in two mammary tumor strains derived from MTW9. Maximum binding to membranes of both tumors occurred at pH 7.6 after incubation for 30 hr at 4 degrees. The binding was inhibited only by polypeptide hormones that possess lactogenic activity. MTW9-P, an ovariectomy-responsive tumor developed in rats maintained on daily perphenazine injections, had 4-fold-higher prolactin binding than had MTW9MtT, an ovariectomy-nonresponsive tumor developed in rats bearing the mammosomatotropic pituitary tumor, MtTW10. Withdrawal of perphenazine from rats bearing MTW9-P caused a fall to normal of plasma prolactin, no tumor regression, and no significant change in prolactin binding. In contrast, resection of MtT resulted in tumor regression, a fall to normal of serum prolactin, and a nearly 3-fold increase in prolactin binding. Scatchard plots of prolactin binding data yield an apparent affinity constant, Ka, of 1.2 X 109 liters/mole for both tumors. The 4-fold-higher prolactin binding in the ovariectomy responsive variant suggests a positive correlation between ovariectomy response and the number of membrane prolactin-binding sites. No correlation between prolactin sensitivity and prolacting binding is apparent.


Mexican Regulation of Biobanks.
Biobank-based research in Mexico is mostly governed by research and data protection laws. There is no direct mention of biobanks in either statutory or regulatory law besides a requirement that the Federal Ministry of Health and a Mexican institution devoted to scientific research approve the transfer of biological materials outside of Mexico for population genetics research purposes. Such requirements are the basis of Genomic Sovereignty in Mexico, but such requirements have not prevented international collaboration. In addition, Mexican law singles out genetic research in informed consent provisions, but it does not specify whether all biobank-based research is genetic research. In order to facilitate international collaboration on biobank-based research, Mexico should directly address biobanking in its laws, building on the research framework and data protection framework already in place.


A School-Based Mobile App Intervention for Enhancing Emotion Regulation in Children: Exploratory Trial.
Most mental health disorders are first experienced in childhood. The rising rates of mental health difficulties in children highlight the need for innovative approaches to supporting children and preventing these difficulties. School-based digital interventions that address shared risk factors and symptoms, such as emotion dysregulation, present exciting opportunities to enhance mental health support for children on a larger scale. This study investigates the use of a new app-based intervention designed to support children's emotion regulation in schools. The aim is to optimize the usability, acceptability, and utility of the app and explore its scope for implementation with the target user in the school context. As part of an interdisciplinary development framework, the app is being evaluated in a 3-month trial across 4 primary schools. In total, 144 children (aged 10-12 years) took part and accessed the intervention app in the classroom or at home. Outcomes regarding usability, acceptability, and implementation opportunities were assessed through digital user data, self-report questionnaires (132/144, 91.6%), and semistructured interviews with children (19/144, 13.2%) and teachers (6/8, 75%). The app usage data showed that 30% (128/426) of the users were returning users. Self-report data indicated that 40.1% (53/132) of the children had not used the app, whereas 57.5% (76/132) had used it once or more. Of the children who had used the app, 67% (51/76) reported that the app was helpful. Interviews with children and teachers suggested positive experiences with the app and that it helped them to calm down and relax. Children reported that they perceived the app as acceptable, usable, and helpful. In terms of the intervention's usability, most features functioned well; however, certain technical issues were reported, which may have led to reduced engagement levels. Teachers not only reported overall positive experiences but also discussed access difficulties and reported a lack of content as one of the main barriers to implementing the app. Having a web-based app significantly enhanced accessibility across devices and settings and provided teachers with more opportunities to use it. We identified the need for new, activating app features in addition to the existing, primarily relaxing ones. The findings indicated that it is possible to use and evaluate an app intervention in the school context and that the app could help enhance children's emotion regulation. We discuss areas for improvement regarding the app, study design, and future implementation strategies. We share important insights with regard to the development, implementation, and evaluation of a new app for supporting children's emotion regulation in schools. Our results demonstrate that mental health apps represent a promising means to facilitate effective mental health service provision in and outside of the school context. Important lessons learned are shared to support other researchers and clinicians on similar journeys.


The impact of parenthood on risk of registration for alcohol use disorder in married individuals: a Swedish population-based analysis.
Although being married with children is associated with a reduced rate of alcohol use disorder (AUD), is this finding independent of a marital effect, different in mothers and fathers and potentially causal in effect. Using Cox proportional hazards, we examined, in 1 252 237 married individuals, the association between a resident younger and older child and risk for AUD registration in national medical, criminal, and pharmacy registers. Using logistic regression, we analyzed, in 600 219 parents, within-person models comparing risk for AUD prior to first pregnancy v. with young children. We examined whether risk for AUD in 1302 parents after a first spousal AUD registration was reduced by having a young resident child. Compared with childless married individuals, resident younger children were associated with a reduced risk for AUD in mothers [hazard ratio (HR) 0.36, 95% confidence interval 0.31-0.41] and fathers (HR 0.66, 0.60-0.73). The reduced risk was attenuated but still significant for older children. Within-person models confirmed the protective effect of young children in mothers [odds ratio (OR) 0.49, 0.30-0.80] but yielded inconclusive results in fathers (OR 0.85, 0.58-1.25). After a first spousal registration for AUD, a resident young child was associated with a substantial reduction in risk for mothers and a weaker marginal effect in fathers. In married individuals, resident children are associated with a reduction in basal risk for AUD which is stronger in mothers than fathers and with younger v. older children. This effect is also evident during high-risk periods. In mothers, our results are consistent with a largely causal effect.


[The effect of caffeine on the electrical reactions of the acinar cells in the canine pancreas].
The action of pharmacological drugs (caffeine, procaine, ruthenium red) which influenced the release of Ca2+ from endoplasmatic reticulum, the electrical responses of dog pancreatic acinar cells was investigated using intracellular glass microelectrodes. These drugs were used to elucidate the mechanisms of Ca2+ release from endoplasmatic reticulum. Membrane depolarization and decrease of input resistance were observed in the presence of caffeine. Procaine and ruthenium red suppressed electrical responses of acinar cells to caffeine. The results obtained permit supposing that there are two mechanisms of Ca2+ release from intracellular stores in pancreatic acinar cells: Ca(2+)-induced one and using inositol-1,4,5-triphosphate.


Influence of diabetes and diabetic retinopathy on the performance of Heidelberg retina tomography II for diagnosis of glaucoma.
To determine whether diabetes and diabetic retinopathy (DR) affect the performance of the Heidelberg Retina Tomograph II (HRT II; Heidelberg Engineering, Heidelberg, Germany) algorithms for glaucoma detection. This population-based survey was conducted among Malays in Singapore who were a 40 to 80 years of age. Diabetes was defined as self-report of a physician's diagnosis, use of diabetic medication, or a random blood glucose level ≥11.1 mmol/L. Retinal photographs were graded for DR according to the modified Airlie House classification system. The diagnosis of glaucoma was based on International Society for Geographical and Epidemiologic Ophthalmology criteria. The sensitivity and the false-positive rates were calculated for the Moorfields regression analysis [MRA]; the linear discriminant functions (LDFs) by Mikelberg (Mikelberg-LDF), Burk (Burk-LDF), and Bathija (Bathija-LDF); and the support vector machine (SVM). A total of 1987 persons without diabetes (including 86 with glaucoma) and 524 with diabetes (including 26 with glaucoma) were analyzed. The presence of diabetes had no influence on both the sensitivities and false-positive rates for all HRT algorithms. In the multivariate analyses adjusting for optic disc size, the presence of DR was significantly associated with the higher false-positive rates for Burk-LDF and Bathija-LDF (P < 0.05), but not with the false-positive rates for MRA, Mikelberg-LDF, and SVM. Diabetes does not affect the performance of HRT II for diagnosis of glaucoma, but the presence of DR may be a source of false-positive test results.


LinMap: visualizing complexity gradients in evolutionary landscapes.
This article describes an interactive visualization tool, LinMap, for exploring the structure of complexity gradients in evolutionary landscapes. LinMap is a computationally efficient and intuitive tool for visualizing and exploring multidimensional parameter spaces. An artificial cell lineage model is presented that allows complexity to be quantified according to several different developmental and phenotypic metrics. LinMap is applied to the evolutionary landscapes generated by this model to demonstrate that different definitions of complexity produce different gradients across the same landscape; that landscapes are characterized by a phase transition between proliferating and quiescent cell lineages where both complexity and diversity are maximized; and that landscapes defined by adaptive fitness and complexity can display different topographical features.


Successes, challenges, and opportunities for quantum chemistry in understanding metalloenzymes for solar fuels research.
Quantum chemical approaches today are a powerful tool to study the properties and reactivity of metalloenzymes. In the field of solar fuels research these involve predominantly photosystem II and hydrogenases, which catalyze water oxidation and hydrogen evolution, as well as related biomimetic and bio-inspired models. Theoretical methods are extensively used to better comprehend the nature of catalytic intermediates, establish important structure-function and structure-property correlations, elucidate functional principles, and uncover the catalytic activity of these complex systems by unravelling the key steps of their reaction mechanism. Computations in the field of water oxidation and hydrogen evolution are used as predictive tools to elucidate structures, explain and synthesize complex experimental observations from advanced spectroscopic techniques, rationalize reactivity on the basis of atomistic models and electronic structure, and guide the design of new synthetic targets. This feature article covers recent advances in the application of quantum chemical methods for understanding the nature of catalytic intermediates and the mechanism by which photosystem II and hydrogenases achieve their function, and points at essential questions that remain only partly answered and at challenges that will have to be met by future advances and applications of quantum and computational chemistry.


Associations of changes in body mass index with all-cause and cardiovascular mortality in healthy middle-aged adults.
Conflicting data exist regarding the association of body mass index (BMI) changes with all-cause and cardiovascular (CV) mortality. The current study investigated the association between changes in BMI and all-cause, CV, and non-CV mortality in a large cohort of middle-aged adults. A total of 379,535 adults over 40 years of age without pre-existing CV disease or cancer at baseline were enrolled to undergo a series of at least three health examinations of biennial intervals. Changes in BMI between baseline, midpoint follow-up, and final health examination during mean 9.3 years were defined according to the pattern of BMI change as follows: stable, sustained gain, sustained loss, and fluctuation. The relationship between BMI change category and mortality was assessed by multivariate Cox regression reporting hazard ratio (HR) with 95% confidence interval (95% CI). During a mean follow-up of 10.7 years for mortality, 12,378 deaths occurred from all causes, of which 2,114 were CV and 10,264 were non-CV deaths. Sustained BMI gain was associated with the lower risk of all-cause (HR 0.89, 95% CI: 0.83-0.95), CV (HR 0.84, 95% CI 0.72-0.98), and non-CV mortality (HR 0.90, 95% CI 0.84-0.96) compared with stable BMI. Conversely, sustained BMI loss (HR 1.25, 95% CI 1.19-1.32) and fluctuation (HR 1.13, 95% CI 1.08-1.19) displayed a higher risk of all-cause mortality compared with stable BMI, which was mainly attributable to the increase in non-CV mortality. Sustained BMI gains were associated with reduced risk of all-cause, CV, and non-CV mortality in middle-aged healthy adults.


A threatening opportunity: The prospect of conversations about race-related experiences between Black and White friends.
Similarities are foundational to building and maintaining friendships, but for cross-race friends, differences in experiences related to race are also inevitable. Little is known about how friends approach talking about race-related experiences. We suggest that these conversations are a threatening opportunity. Across five studies, we show that they can enhance closeness and intergroup learning among Black and White friends but that these benefits can be accompanied, and sometimes prevented by identity threat. In Study 1, Black (N = 57) and White (N = 59) adults anticipated both benefits and risks of such conversations, though more benefits than risks. In Study 2A (N = 143) and Study 2B (N = 149), Black participants reported less willingness to disclose race-related experiences to extant White friends than Black friends and anticipated feeling less comfortable doing so, controlling for closeness. However, they also desired to be understood by Black and White friends equally. In Study 3 (N = 147) and Study 4 (N = 172), White participants also felt less comfortable when an imagined Black friend disclosed race-related versus nonrace-related experiences to them. However, they felt closer to their friend after the race-related disclosure. Additionally, they felt more comfortable hearing about race-related experiences from a friend than through a third party and they reported learning more when the race-related experience was a friend's than a stranger's. Taken together, the studies highlight the benefits as well as the risks of conversations about race for cross-race friends and the need for future studies that track real-time conversations and test strategies to help friends engage in these conversations productively. (PsycInfo Database Record (c) 2022 APA, all rights reserved).


[Kidney tubule dysfunction caused by acetic acid].
Two patients admitted after ingestion of 80% acetic acid are described. Only the first patient developed haemolysis, slight intravascular coagulation and oliguric kidney insufficiency. They were treated with a nasogastric tube and total parenteral feeding. During the first week after admission urinary excretion of beta 2-microglobulin, alanine-aminopeptidase and N-acetyl-glucosaminidase was significantly increased. The patients remained haemodynamically stable and did not develop fever. The above-mentioned elevated excretions returned to normal levels. Both patients showed similar patterns of tubular proteinuria. The observations in the second patient suggest a direct toxic effect of acetic acid on the proximal tubule of the kidney.


The virtual doppelganger: effects of a virtual reality simulator on perceptions of schizophrenia.
Recent scholarship suggests that virtual environments can serve as effective proxies in battling implicit stereotypes. However, existing experimental research has rarely examined the effectiveness of virtual simulations of mental illnesses in inducing empathy to combat stereotypical responses. We report results from a 4-condition, between subjects experiment (N = 112), wherein participants were exposed to either a virtual simulation of schizophrenia, a written empathy-set induction of schizophrenia, a combination of both the simulation and written empathy conditions, or a control condition. The results indicated that the virtual simulation + empathy condition induced greater empathy and more positive perceptions toward people suffering from schizophrenia than the control or written empathy-set condition. Interestingly, the simulation-only condition resulted in the greatest desire for social distance whereas not significantly differing on empathy and attitude measures from either the written empathy or simulation + empathy conditions. We discuss the implications of the findings and recommend directions for future research.


Transient alginate gene expression by Pseudomonas putida biofilm residents under water-limiting conditions reflects adaptation to the local environment.
Under water-limiting conditions Pseudomonas putida produces the exopolysaccharide alginate, which influences biofilm development and facilitates maintaining a hydrated microenvironment. Since alginate is a minor biofilm matrix component it is important to determine whether alginate production occurs by all or a subset of residents, and when and to what extent cells contribute to alginate production. To address these questions we employed stable and unstable fluorescent reporters to measure alginate biosynthesis (algD) operon expression and metabolic activity in vivo quantitatively by flow cytometry and visually by microscopy. Here we report that during growth under water-limiting conditions and when biofilms become dehydrated most residents transiently express the alginate biosynthesis genes leading to distinct spatial patterns as the biofilm ages. Transient alginate gene expression was not a consequence of decreased metabolic activity, since metabolic reporters were still expressed, nor was it likely due to transient cytosolic availability of the alternative sigma factor AlgT, based on qRT-PCR. Our findings also indicate that one or more biofilm attribute, other than alginate, provides protection from desiccation stress. Collectively, our findings suggest that differentiated cells dedicated to alginate production are not part of the P. putida biofilm lifestyle under water-limiting conditions. Alternatively, P. putida biofilm cells may be responding to their own local environment, producing alginate because of the fitness advantage it confers under those particular conditions.


Persistent postural-perceptual dizziness (PPPD): a common, characteristic and treatable cause of chronic dizziness.
Persistent postural-perceptual dizziness (PPPD) is a newly defined diagnostic syndrome that unifies key features of chronic subjective dizziness, phobic postural vertigo and related disorders. It describes a common chronic dysfunction of the vestibular system and brain that produces persistent dizziness, non-spinning vertigo and/or unsteadiness. The disorder constitutes a long-term maladaptation to a neuro-otological, medical or psychological event that triggered vestibular symptoms, and is usefully considered within the spectrum of other functional neurological disorders. While diagnostic tests and conventional imaging usually remain negative, patients with PPPD present in a characteristic way that maps on to positive diagnostic criteria. Patients often develop secondary functional gait disorder, anxiety, avoidance behaviour and severe disability. Once recognised, PPPD can be managed with effective communication and tailored treatment strategies, including specialised physical therapy (vestibular rehabilitation), serotonergic medications and cognitive-behavioural therapy.


Night-time feeding of Bmal1-/- mice restores SCFA rhythms and their effect on ghrelin.
The known crosstalk between short-chain fatty acids (SCFAs) and the circadian clock is tightly intertwined with feeding time. We aimed to investigate the role of the core clock gene Bmal1 and feeding time in the diurnal rhythms in plasma and caecal SCFA levels and in their effect on the release of the hunger hormone ghrelin in the stomach and colon. WT, Bmal1-/- (ad libitum fed) and night-time-restricted-fed (RF)-Bmal1-/- littermates were killed at zeitgeber time (ZT) 4 and 16. SCFA concentrations were measured by gas chromatography. To investigate the effect of SCFAs on ghrelin release, stomach and colonic full-thickness strips were incubated with Krebs or a SCFA mix mimicking plasma or caecal concentrations, after which octanoyl ghrelin release was measured by RIA. Diurnal rhythms in caecal and plasma SCFAs oscillated in phase but rhythmic changes were abolished in Bmal1-/- mice. RF of Bmal1-/- mice restored fluctuations in caecal SCFAs. Plasma SCFA concentrations failed to affect gastric ghrelin release. The effect of caecal SCFA concentrations on colonic ghrelin release was rhythmic (inhibition at ZT 4, no effect at ZT 16). In Bmal1-/- mice, the inhibitory effect of SCFAs at ZT 4 was abolished. RF Bmal1-/- mice restored the inhibitory effect and increased colonic Clock expression. To conclude, diurnal fluctuations in caecal SCFAs and the effect of SCFAs on colonic ghrelin release are regulated by feeding time, independent of the core clock gene Bmal1. However, local entrainment of other clock genes might contribute to the observed effects.


In situ FTIR study of CO2 reduction on inorganic analogues of carbon monoxide dehydrogenase.
The CO2-to-CO reduction by carbon monoxide dehydrogenase (CODH) with a [NiFe4S4] cluster is considered to be the oldest pathway of biological carbon fixation and therefore may have been involved in the origin of life. Although previous studies have investigated CO2 reduction by Fe and Ni sulfides to identify the prebiotic origin of the [NiFe4S4] cluster, the reaction mechanism remains largely elusive. Herein, we applied in situ electrochemical ATR-FTIR spectroscopy to probe the reaction intermediates of greigite (Fe3S4) and violarite (FeNi2S4). Intermediate species assignable to surface-bound CO2 and formyl groups were found to be stabilized in the presence of Ni, lending insight into its role in enhancing the multistep CO2 reduction process.


Modulation of nerve growth factor in peripheral organs by estrogen and progesterone.
Nerve growth factor (NGF) synthesized in peripheral organs plays a critical role in the development and maintenance of the nervous system and also participates in processing nociceptive stimuli. Previous studies suggest that reproductive hormones may regulate the expression of NGF. Ovariectomies were performed on female mice, and mice were killed 24 h after hormone replacement to evaluate the effects of estrogen and progesterone on NGF in peripheral organs, specifically the uterus, bladder, heart, and salivary gland. Sham-operated intact mice and untreated ovariectomized mice served as controls. Immunohistochemistry demonstrated the presence of NGF, estrogen receptor-alpha, estrogen receptor-beta, and progesterone receptors in these organs. Ovariectomy caused a significant decrease in NGF protein content in the uterus, and short term treatment of ovariectomized mice with estrogen and/or progesterone increased uterine NGF mRNA and restored NGF protein to concentrations similar to intact control mice. Ovariectomy did not affect NGF protein concentrations in the salivary gland, but treatment of ovariectomized mice with estrogen alone or in conjunction with progesterone stimulated concentrations of NGF protein that exceeded those observed in intact control or ovariectomized, untreated mice. NGF mRNA was increased in salivary glands from ovariectomized mice treated with progesterone alone or in combination with estrogen relative to other groups. NGF protein content of the hearts of ovariectomized mice treated with estrogen alone or in conjunction with progesterone was increased relative to intact controls and ovariectomized, untreated mice, but neither ovariectomy or hormone replacement affected NGF mRNA content in the heart. NGF protein content of the bladder was unaffected by ovariectomy or hormone treatment, and bladder NGF mRNA was unaffected by ovariectomy or hormone treatment. Collectively, these results indicate that reproductive hormones have the capacity to regulate NGF message and protein in a manner that varies among organs. Fluctuations in the expression of NGF, in conjunction with other factors, may help to explain gender differences in pain sensation and inflammatory response.


Reconstruction after median sternotomy infection.
Reconstruction after median sternotomy infections is a difficult problem. We have presented ten consecutive patients with a wide variety of underlying cardiac illnesses who have had this dreaded complication develop. All of these patients have undergone successful reconstruction after adequate debridement and elimination of the mediastinal dead space using a combination of well vascularized omentum and pectoralis major muscle flaps. Stability of the chest wall has been accomplished primarily with pectoralis major muscle flaps. Complications have been few, morbidity has been significantly reduced and mortality, thus far, has been eliminated, although we certainly have no expectations that it will remain so.


High molecular weight, but not total, CTRP3 levels are associated with serum triglyceride levels.
C1q/TNF-related protein 3 (CTRP3) is a relatively novel adipose tissue-derived cytokine (adipokine) which has been linked to improved glucose regulation and insulin sensitivity. However, the relationship between circulating CTRP3 levels and diabetes is controversial. CTRP3 can circulate in different oligomeric complexes: trimeric, hexameric, and high molecular weight (HMW) oligomeric complexes. However, the concentration of the different oligomeric complexes in human disease states has not been previously investigated. Therefore, the purpose of this study was to compare the levels of different oligomeric complexes of CTRP3 between type 2 diabetic and nondiabetic individuals. Additionally, the association between the oligomeric complexes and other serum factors was examined. CTRP3 primarily circulates in the HMW complex (>50%) and the hexametric multimer, with no CTRP3 detected in the trimeric complex or as a monomer. Further, no differences were observed in total, hexameric, or HMW CTRP3 levels regardless of diabetic status. Surprisingly, HMW CTRP3 was found to be positively correlated with circulating triglyceride levels. Combined, these data suggest that CTRP3 is associated with triglyceride regulation, not diabetic status. These data may explain some of the discrepancies in the literature as elevated triglyceride levels are often detected in patients with obesity and type 2 diabetes.


Gene expression and immunolocalisation of amelogenins in developing embryonic and neonatal hamster teeth.
Amelogenins are a group of related matrix proteins, synthesised and secreted by ameloblasts during the formation of dental enamel. We have examined expression patterns and the tissue distribution of amelogenins by in situ hybridisation and by immunohistochemistry of developing teeth of embryonic (E12-E15) and neonatal (1- to 4-day-old) golden hamsters. Amelogenin expression and (intracellular) immunostaining for amelogenins were first observed in late embryonic stages in E14 incisors and E15 first molars in partially polarised pre-ameloblasts located along a thin layer of predentine before any overt deposition of enamel. Expression of mRNA and protein staining for amelogenins increased with age and early pre-dentine became immunopositive. The highest mRNA levels and substantial immunostaining for amelogenins were noted in neonatal-stage secretory ameloblasts fully engaged in enamel matrix deposition. After completion of the secretory phase, amelogenin gene expression continued at a lower level in post-secretory stages and was seen in transition-phase and maturation-phase ameloblasts. No amelogenin transcripts were observed in odontoblasts at any stage of their development. However, young odontoblasts stained weakly with anti-amelogenin antibodies before they formed the first layer of dentine, although this staining disappeared in odontoblasts at later stages of development. We conclude that amelogenin gene transcription occurs as early as the polarisation stage of pre-ameloblasts and is closely followed by translation of mRNA into amelogenin proteins. Odontoblasts do not transcribe the amelogenin gene and probably endocytose and digest amelogenins from the pre-dentine. Amelogenins are also transcribed but at a low level in post-secretory stages of amelogenesis.


Effect of phosphate availability on biofilm formation in cooling towers.
Phosphate limitation has been suggested as a preventive method against biofilms. P-limited feed water was studied as a preventive strategy against biofouling in cooling towers (CTs). Three pilot-scale open recirculating CTs were operated in parallel for five weeks. RO permeate was fed to the CTs (1) without supplementation (reference), (2) with supplementation by biodegradable carbon (P-limited) and (3) with supplementation of all nutrients (non-P-limited). The P-limited water contained ≤10 µg PO4 l-1. Investigating the CT-basins and coupons showed that P-limited water (1) did not prevent biofilm formation and (2) resulted in a higher volume of organic matter per unit of active biomass compared with the other CTs. Exposure to external conditions and cycle of concentration were likely factors that allowed a P concentration sufficient to cause extensive biofouling despite being the limiting compound. In conclusion, phosphate limitation in cooling water is not a suitable strategy for CT biofouling control.


Pregnancy, cytokines, and disease activity in systemic lupus erythematosus.
To evaluate levels of selected cytokines and soluble receptors involved in the humoral immune response during pregnancy in systemic lupus erythematosus (SLE) patients. Seventeen consecutive SLE patients and 8 matched healthy controls were prospectively studied during pregnancy. Sera were obtained within the last 3 months prior to pregnancy; at 9, 17, and 29 weeks of pregnancy; and at 1 month after delivery. Serum levels of interleukin-10 (IL-10), interleukin-6 (IL-6), and soluble tumor necrosis factor receptors p55 (sTNFR I) and p75 (sTNFR II) were evaluated. SLE activity was measured by the European Consensus Lupus Activity Measurement score modified for pregnancy. IL-10 serum levels were found to be higher (P <0.0001) in patients than in controls before conception, and still higher (P <0.0001) in SLE patients during gestation, without intertrimester changes. In SLE patients, IL-6 serum levels did not increase in the third trimester of pregnancy, as was observed in controls (P=0.011). No significant differences between SLE patients and controls were found in either sTNFR I or II levels or profiles before and during pregnancy. IL-10 and sTNFR I levels were significantly higher during pregnancy and postpartum in SLE patients with active disease (P=0.03 and P=0.01, respectively). The levels of some cytokines involved in the humoral immune response seem to be modified in the peripheral circulation of pregnant SLE patients. The most relevant modifications are the lower than expected increase of IL-6 in the third trimester of gestation and persistently high levels of IL-10 during pregnancy.


Single-Cell and CellChat Resolution Identifies Collecting Duct Cell Subsets and Their Communications with Adjacent Cells in PKD Kidneys.
ADPKD is a genetic disorder with a molecular complexity that remains poorly understood. In this study, we sampled renal cells to construct a comprehensive and spatiotemporally resolved gene expression atlas in whole Pkd1 mutant polycystic mouse kidneys at single-cell resolution. We characterized cell diversity and identified novel collecting duct (CD) cell subtypes in cystic kidneys. We further found that CD cells appear to take different cell fate trajectories, and the first and the most important step might take place around day 14 in Pkd1 homozygous kidneys. After that day, increased numbers of CD cells showed highly proliferative and fibrotic characteristics, as detected in later-stage Pkd1 homozygous kidneys, both of which should contribute to cyst growth and renal fibrosis. With a newly developed modeling algorithm, called CellChat Explorer, we identify cell-to-cell communication networks mediated by the ligand receptor, such as MIF-CD44/CD74, in cystic kidneys, and confirm them via the expression patterns of ligands and receptors in four major cell types, which addresses the key question as to whether and how Pkd1 mutant renal epithelial cells affect their neighboring cells. The allele-specific gene expression profiles show that the secretion of cytokines by Pkd1 mutant epithelial cells may affect the gene expression profiles in recipient cells via epigenetic mechanisms, and vice versa. This study can be used to drive precision therapeutic targeting of ADPKD.


Direct Demonstration That Loop1 of Scap Binds to Loop7: A CRUCIAL EVENT IN CHOLESTEROL HOMEOSTASIS.
Cholesterol homeostasis is mediated by Scap, a polytopic endoplasmic reticulum (ER) protein that transports sterol regulatory element-binding proteins from the ER to Golgi, where they are processed to forms that activate cholesterol synthesis. Scap has eight transmembrane helices and two large luminal loops, designated Loop1 and Loop7. We earlier provided indirect evidence that Loop1 binds to Loop7, allowing Scap to bind COPII proteins for transport in coated vesicles. When ER cholesterol rises, it binds to Loop1. We hypothesized that this causes dissociation from Loop7, abrogating COPII binding. Here we demonstrate direct binding of the two loops when expressed as isolated fragments or as a fusion protein. Expressed alone, Loop1 remained intracellular and membrane-bound. When Loop7 was co-expressed, it bound to Loop1, and the soluble complex was secreted. A Loop1-Loop7 fusion protein was also secreted, and the two loops remained bound when the linker between them was cleaved by a protease. Point mutations that disrupt the Loop1-Loop7 interaction prevented secretion of the Loop1-Loop7 fusion protein. These data provide direct documentation of intramolecular Loop1-Loop7 binding, a central event in cholesterol homeostasis.


Investigating the temporal relationship between proactive burnout prevention and burnout: A four-wave longitudinal study.
Given the detrimental effects of burnout for individuals and organizations, it is of crucial importance to better understand the self-initiated actions employees take to prevent burnout. While such proactive burnout prevention is likely to reduce burnout complaints, these activities may also be frustrated by high burnout levels. This means that proactive burnout prevention and burnout can negatively affect each other over time. The present study used a four-wave longitudinal panel design to investigate temporal relationships between proactive burnout prevention and burnout over 3, 6 and 9 weeks. Participants were 165 employees in the financial services industry who provided data on all four measurement occasions. The outcomes of structural equation modelling provided support for the hypothesized combined effects model compared to the lagged and reversed effects models. The findings suggest that proactive burnout prevention can help to prevent burnout, while engagement in these behaviours may be hindered by high initial levels of burnout. Employees should therefore intervene before their resource pool becomes too depleted and they lack the energy or mental strength to invest resources, in order to proactively retain or regain resources.


Neurovascular bundle decompression without excessive dissection for tarsal tunnel syndrome.
Tarsal tunnel syndrome (TTS) is an entrapment neuropathy of the posterior tibial nerve and its branches in the tarsal tunnel. We present our less invasive surgical treatment of TTS in 69 patients (116 feet) and their clinical outcomes. The mean follow-up period was 64.6 months. With the patient under local anesthesia we use a microscope to perform sharp dissection of the flexor retinaculum and remove the connective tissues surrounding the posterior tibial nerve and vessels. To prevent postoperative adhesion and delayed neuropathy, decompression is performed to achieve symptom improvement without excessive dissection. Decompression is considered complete when the patient reports intraoperative symptom abatement and arterial pulsation is sufficient. The sensation of numbness and/or pain and of foreign substance adhesion was reduced in 92% and 95% of our patients, respectively. In self-assessments, 47 patients (68%) reported the treatment outcome as satisfactory, 15 (22%) as acceptable, and 7 (10%) were dissatisfied. Of 116 feet, 4 (3%) required re-operation, initial decompression was insufficient in 2 feet and further decompression was performed; in the other 2 feet improvement was achieved by decompression of the distal tarsal tunnel. Our surgical method involves neurovascular bundle decompression to obtain sufficient arterial pulsation. As we use local anesthesia, we can confirm symptom improvement intraoperatively, thereby avoiding unnecessary excessive dissection. Our method is simple, safe, and without detailed nerve dissection and it prevents postoperative adhesion.


Characterization of the yerba mate (Ilex paraguariensis) volatile fraction using solid-phase microextraction-comprehensive 2-D GC-MS.
The present research is focused on the use of a solid-phase microextraction-comprehensive 2-D GC methodology, in the analysis of the volatile fraction of yerba mate. Yerba mate is used for the generation of a tea-like beverage, widely consumed in South America. A rapid-scanning quadrupole mass spectrometer (qMS), employed as a detection system and operated at a 25 Hz scanning frequency, supplied high-quality mass spectra. The effectiveness of the 3-D comprehensive 2-D GC-qMS experiment was compared to that of GC-qMS analysis on the same sample. Peak identification, in both applications, was achieved through MS library matching, with the support of linear retention index data. Apart from a great increase in the number of analytes separated (approx. by a factor of 5) and identified (approx. by a factor of 3.5), the comprehensive 2-D GC-qMS approach enabled the determination of a high number of hazardous contaminants (aliphatic hydrocarbons, polycyclic aromatic hydrocarbons, and plasticizers), barely visible in the GC-qMS analysis.


Adenomyosis in Pakistani women: four year experience at the Aga Khan University Medical Centre, Karachi.
As part of a quality assurance programme at the Aga Khan University Medical Centre, Karachi, Pakistan, all hysterectomy specimens were reviewed from January 1986 to December 1989. Adenomyosis was found in 237 of the 419 (56.5%) specimens studied. Of these 237 patients, 232 (97.9%) were parous and 196 (82.8%) were in the fourth and fifth decades of life. This high prevalence in parous women aged 40-59 years was significant. Fibroids, cervicitis, and endometrial hyperplasia were the most common associated diagnoses. Of all the associations studied, only endometrial hyperplasia was significantly more prevalent in the group with adenomyosis. Adenomyosis was stated as an indication for surgery in 69 patients and was confirmed by histopathology in 49 (71%). Preoperative suspicion of adenomyosis was present in 49 (20.6%) patients of all those ultimately found to have the disease. There is a high prevalence of adenomyosis in the population studied, which indicates that the condition may have been underdiagnosed in the past, especially as it is difficult to diagnose without surgery and hysterectomy is currently the only treatment.


Use of prescription drugs in the older adult population-a nationwide pharmacoepidemiological study.
Multi-morbidity and polypharmacy are common among older people. It is essential to provide a better understanding of the complexity of prescription drug use among older adults to optimise rational pharmacotherapy. Population-based utilisation data in this age group is limited. Using the Danish nationwide health registries, we aimed to characterise drug use among Danish individuals ≥ 60 years. This is a descriptive population-based study assessing drug prescription patterns in 2015 in the full Danish population aged ≥ 60 years. The use of specific therapeutic subgroups and chemical subgroups and its dependence on age were described using descriptive statistics. Profiles of drug combination patterns were evaluated using latent class analysis. We included 1,424,775 residents (median age 70 years, 53% women). Of all the older adults, 89% filled at least one prescription during 2015. The median number of drug groups used was five per person. The most used single drug groups were paracetamol and analogues (34%), statins (33%) and platelet aggregation inhibitors (24%). Eighteen drug profiles with different drug combination patterns were identified. One drug profile with expected use of zero drugs and 11 drug profiles expected to receive more than five different therapeutic subgroup drugs were identified. The use of drugs is extensive both at the population level and increasing with age at an individual level. Separating the population into different homogenous groups related to drug use resulted in 18 different drug profiles, of which 11 drug profiles received on average more than five different therapeutic subgroup drugs.


An Organ for My Child: Public Appeals for Limited Resources.
The case of Jamie Fiske and subsequent public appeals for particular children by President Ronald Reagan represent classic cases in pediatric bioethics in which parents or others publicly appealed for a donor organ for a particular child. These raise questions about the appropriate boundaries for public appeals for a limited resource for a particular child and how the press and medical community should respond to such appeals. Public appeals by parents to advocate for their child to receive a limited resource above others promote rationing by morally irrelevant factors and shift the public focus from the national shortage of organs for transplant to the needs of a particular child. Yet these appeals are understandable and will likely continue. Recognizing this, we consider appropriate responses by the media, transplant community, hospitals, and individual clinicians.


Is patellectomy compatible with an Army career?
The aim of this study was to assess the impact of patellectomy on the service career of soldiers. Information on 184 individuals on whom patellectomy was performed between 1981 and 1986, was retrieved from computer records. The criterion for success of the operation was the recovery to a fully fit category. Fifty three patients (29%) regained full fitness. The outcome of patellectomies performed for chondromalacia patellae and osteoarthritis appears to be worse than that for patellar fractures.


Enhanced patterns on intraoperative contrast-enhanced ultrasonography predict outcomes after curative liver resection in patients with hepatocellular carcinoma.
This study aimed to clarify what hepatocellular carcinoma (HCC) phenotype, as categorized by intraoperative contrast-enhanced ultrasonography (CEUS), showed a high risk of recurrence after hepatic resection. Patients who underwent initial curative hepatectomy with intraoperative CEUS for a single HCC nodule were retrospectively assigned to three patterns of fine (FI), vascular (VA), and irregular (IR) according to the maximum intensity projection pattern based on intraoperative CEUS. Staining was performed for Ki-67, pyruvate kinase type M2 (PKM2), and vascular endothelial growth factor (VEGF) to assess the tumor proliferative activity, tumor glucose metabolism, and angiogenesis, respectively. Of 116 patients, 18, 50, and 48 were assigned to the FI, VA and IR patterns, respectively. IR patients demonstrated a significantly worse prognosis for both the recurrence-free survival (RFS) and overall survival (OS) (P = 0.0002, 0.0262, respectively) than did patients with other patterns. A multivariate analysis revealed an IR pattern in intraoperative CEUS to be an independent predictive factor for a poor RFS, and major hepatectomy and an IR pattern were independent predictive factors for a poor OS. An IR pattern was closely related to the tumor size (≥ 3.3 cm) and poor histological differentiation and showed a high Ki-67 index, low VEGF expression, and high PKM2 expression. IR-pattern HCCs as classified by intraoperative CEUS may be associated with a higher risk of recurrence and worse outcomes in HCC patients after hepatic resection than other patterns.


A review of the virological efficacy of the 4 World Health Organization-recommended tenofovir-containing regimens for initial HIV therapy.
We systematically reviewed studies of the virological efficacy of the 4 new tenofovir (TDF)-containing regimens recommended for initial antiretroviral (ARV) therapy in the 2010 World Health Organization ARV Treatment Guidelines. Thirty-three studies assessed the efficacy of 1 or more TDF-containing regimens: TDF/lamivudine (3TC)/nevirapine (NVP) (n = 3), TDF/ emtricitabine (FTC)/NVP (n = 9), TDF/3TC/efavirenz (EFV) (n = 6), and TDF/FTC/EFV (n = 19). TDF/3TC/NVP was the least well-studied and appeared the least efficacious of the 4 regimens. In 2 comparative studies, TDF/3TC/NVP was associated with significantly more virological failure than AZT/3TC/NVP; a third study was terminated prematurely because of early virological failure. TDF/FTC/NVP was either equivalent or inferior to its comparator arms. TDF/3TC/EFV was equivalent to its comparator arms. TDF/FTC/EFV was equivalent or superior to its comparator arms. Possible explanations for these findings include the greater antiviral activity of EFV versus NVP and longer intracellular half-life of FTC-triphosphate versus 3TC-triphosphate. Further study of TDF/3TC/NVP is required before it is widely deployed for initial ARV therapy.


Influence of airway pressure waveform on cardiac output during positive pressure ventilation of healthy newborn dogs.
During positive pressure ventilation, increases in mean airway pressure produced by increases in end expiratory pressure are associated with reductions in cardiac output. Mean airway pressure may be increased not only by increasing end expiratory pressure, but also by increasing the inspiratory pressure or the inspiratory to expiratory time ratio. During positive pressure ventilation of 10 healthy newborn dogs, cardiac output and left ventricular dimensions were measured using radioactive microspheres and echocardiography, respectively. Baseline ventilation was produced using the least inspiratory pressure to maintain normocarbia (rate = 50 min-1, inspiratory to expiratory time ratio = 1:3, end expiratory pressure = 2 cm H2O). The mean airway pressure was increased 3-fold by independently changing the airway pressure waveform through increases in inspiratory pressure, inspiratory to expiratory time ratio, or end expiratory pressure. Despite differences in airway pressure waveform, similar reductions in left ventricular dimension, cardiac output, and stroke volume were seen. We conclude that at a given increased mean airway pressure during positive pressure ventilation, the reduction in cardiac output is independent of the airway pressure waveform.


Irradiance responsivity and unequivocal type-1 phase responsivity of rat circadian activity rhythms.
Behavioral, neuropharmacological, and molecular studies of light-induced phase shifting of rodent circadian rhythms evaluate carefully the phase dependence of light responsivity. However, much less information is available regarding the dependence of such effects on the duration and irradiance of the photic stimulus used. In this study, very brief (5-min) white light pulses of 50 microW/cm2 (175 lux) administered to hooded rats elicited unequivocal type-1 phase responsivity, without significant changes in period, and with phase shift variability comparable to that in studies using longer, higher intensity pulses. Irradiance dependence was demonstrated in the phase-delay, phase-advance, and crossover portions of the phase-response curve, with minimal phase shifting seen during the dead zone even at very high irradiance. These results indicate that maximal phase-shifting magnitude may be achieved with shorter, less intense photic stimuli than are often used in studies of the neural mechanisms involved in light responsivity of rat circadian rhythms.


Deep learning and radiomics: the utility of Google TensorFlow™ Inception in classifying clear cell renal cell carcinoma and oncocytoma on multiphasic CT.
Currently, all solid enhancing renal masses without microscopic fat are considered malignant until proven otherwise and there is substantial overlap in the imaging findings of benign and malignant renal masses, particularly between clear cell RCC (ccRCC) and benign oncocytoma (ONC). Radiomics has attracted increased attention for its utility in pre-operative work-up on routine clinical images. Radiomics based approaches have converted medical images into mineable data and identified prognostic imaging signatures that machine learning algorithms can use to construct predictive models by learning the decision boundaries of the underlying data distribution. The TensorFlow™ framework from Google is a state-of-the-art open-source software library that can be used for training deep learning neural networks for performing machine learning tasks. The purpose of this study was to investigate the diagnostic value and feasibility of a deep learning-based renal lesion classifier using open-source Google TensorFlow™ Inception in differentiating ccRCC from ONC on routine four-phase MDCT in patients with pathologically confirmed renal masses. With institutional review board approval for this 1996 Health Insurance Portability and Accountability Act compliant retrospective study and a waiver of informed consent, we queried our institution's pathology, clinical, and radiology databases for histologically proven cases of ccRCC and ONC obtained between January 2000 and January 2016 scanned with a an intravenous contrast-enhanced four-phase renal mass protocol (unenhanced (UN), corticomedullary (CM), nephrographic (NP), and excretory (EX) phases). To extract features to be used for the machine learning model, the entire renal mass was contoured in the axial plane in each of the four phases, resulting in a 3D volume of interest (VOI) representative of the entire renal mass. We investigated thirteen different approaches to convert the acquired VOI data into a set of images that adequately represented each tumor which was used to train the final layer of the neural network model. Training was performed over 4000 iterations. In each iteration, 90% of the data were designated as training data and the remaining 10% served as validation data and a leave-one-out cross-validation scheme was implemented. Accuracy, sensitivity, specificity, positive (PPV) and negative predictive (NPV) values, and CIs were calculated for the classification of the thirteen processing modes. We analyzed 179 consecutive patients with 179 lesions (128 ccRCC and 51 ONC). The ccRCC cohort had a mean size of 3.8 cm (range 0.8-14.6 cm) and the ONC cohort had a mean lesion size of 3.9 cm (range 1.0-13.1 cm). The highest specificity and PPV (52.9% and 80.3%, respectively) were achieved in the EX phase when we analyzed the single mid-slice of the tumor in the axial, coronal and sagittal plane, and when we increased the number of mid-slices of the tumor to three, with an accuracy of 75.4%, which also increased the sensitivity to 88.3% and the PPV to 79.6%. Using the entire tumor volume also showed that classification performance was best in the EX phase with an accuracy of 74.4%, a sensitivity of 85.8% and a PPV of 80.1%. When the entire tumor volume, plus mid-slices from all phases and all planes presented as tiled images, were submitted to the final layer of the neural network we achieved a PPV of 82.5%. The best classification result was obtained in the EX phase among the thirteen classification methods tested. Our proof of concept study is the first step towards understanding the utility of machine learning in the differentiation of ccRCC from ONC on routine CT images. We hope this could lead to future investigation into the development of a multivariate machine learning model which may augment our ability to accurately predict renal lesion histology on imaging.


Efficacy of total lymphoid irradiation in intractable rheumatoid arthritis. A double-blind, randomized trial.
Twenty-six patients participated in a randomized, double-blind study of the efficacy of total lymphoid irradiation in the treatment of intractable rheumatoid arthritis. All 26 patients, for whom therapy with gold compounds and penicillamine had failed, would ordinarily have been considered candidates for cytotoxic or antimetabolite drug therapy. Thirteen patients randomly assigned to receive full-dose total lymphoid irradiation (2000 rad) and 11 patients assigned to receive control low-dose total lymphoid irradiation (200 rad) completed radiotherapy. Alleviation of joint disease activity was significantly greater in the high-dose group as judged by morning stiffness, joint tenderness, and functional assessment (global composite score) at 3 and 6 months after radiotherapy. The high-dose group had a marked reduction in both T-lymphocyte function and numbers, but this finding was not observed in the low-dose group. Complications seen in the high-dose but not low-dose group included transient neutropenia, thrombocytopenia, pericarditis, and pleurisy.


Can the COPD web be used to promote self-management in patients with COPD in swedish primary care: a controlled pragmatic pilot trial with 3 month- and 12 month follow-up.
Evaluate the feasibility of the COPD Web and its study design and study procedures and to increase the understanding of the potential effect of the tool in order to provide guidance for a future large scale trial. Parallel-group controlled pragmatic pilot trial. There was a total of 83 patients with COPD (mean age 70 ± 8 years with a forced expiratory volume in first second percent predicted of 60 ± 17%). The intervention group (n = 43) was introduced to and had access to the COPD Web in addition to usual care, while the control group (n = 40) received usual care alone. The feasibility of the COPD Web (i.e., if and how the COPD Web was used) was automatically collected through the website, while outcomes on health, conceptual knowledge, and physical activity (PA) were collected through questionnaires at baseline, 3 months and 12 months. At 3 months, 77% of the intervention group was considered users, and the majority of time spent on the site was related to PA and exercises and was spent during the first month (>80%). In addition, the intervention group reported increased PA (odds ratio [OR] = 4.4, P < .001), increased conceptual knowledge in five domains (OR = 2.6-4.2, all P < .05), and altered disease management strategies (e.g., increased PA) (OR ≥ 2.7 P < .05) in comparison to the control group. The latter was also different between groups at 12 months (OR = 3.7, P = .044). Knowledge of PA was correlated with level of PA (ρ = .425-.512, P < .05) as well as to the use of PA as a strategy to manage their disease (χ2 = 11.2-32.9, P < .05). Giving patients with COPD access to the COPD Web in addition to their ordinary primary care might be an effective shorter term (3 month) strategy to promote self-management. However, these results needs to be confirmed in a definitive large-scale trial. Key points Even though self-management strategies are an important part of chronic obstructive pulmonary disease (COPD) management, access to support for such strategies are limited for a large part of the COPD-population. Promoting self-management through the COPD Web might increase short-term levels of physical activity, promote conceptual knowledge and alter disease management strategies. The primary care COPD population in this study experienced limited impact of the disease in daily life, limited exertional dyspnea, and high generic quality-of-life, but vastly reduced levels of physical activity. A future large scale study should include strategies to encourage greater exposures to the COPD Web, including an extended analysis of factors associated with using or not using the tool over time and its impact on outcome measures, objective measures of conceptual knowledge, and physical activity, and it should include a large enough sample size to enable sub-group analyses and strategies to enhance recruitment.


γδ-Enriched CAR-T cell therapy for bone metastatic castrate-resistant prostate cancer.
Immune checkpoint blockade has been largely unsuccessful for the treatment of bone metastatic castrate-resistant prostate cancer (mCRPC). Here, we report a combinatorial strategy to treat mCRPC using γδ-enriched chimeric antigen receptor (CAR) T cells and zoledronate (ZOL). In a preclinical murine model of bone mCRPC, γδ CAR-T cells targeting prostate stem cell antigen (PSCA) induced a rapid and significant regression of established tumors, combined with increased survival and reduced cancer-associated bone disease. Pretreatment with ZOL, a U.S. Food and Drug Administration-approved bisphosphonate prescribed to mitigate pathological fracture in mCRPC patients, resulted in CAR-independent activation of γδ CAR-T cells, increased cytokine secretion, and enhanced antitumor efficacy. These data show that the activity of the endogenous Vγ9Vδ2 T cell receptor is preserved in CAR-T cells, allowing for dual-receptor recognition of tumor cells. Collectively, our findings support the use of γδ CAR-T cell therapy for mCRPC treatment.


Modified external needle drainage procedure for rhegmatogenous retinal detachment.
To describe the results of a modified external needle subretinal fluid drainage technique to treat eyes with a rhegmatogenous retinal detachment. One hundred eighty-seven consecutive patients with a rhegmatogenous retinal detachment who underwent scleral buckle and modified external needle drainage at Duke Eye Center or Vistar Eye Associates were included in this study. Subretinal fluid was drained by a technique whereby the intraocular pressure was raised to a supranormal level by tightening an encircling scleral buckle and then the subretinal space was entered by a needle introduced externally while the retina was directly viewed with an indirect ophthalmoscope. Subretinal fluid drainage success rate, one-operation reattachment rate, final retinal reattachment rate, intraoperative and postoperative complication rates, and final visual acuity were the main outcome measures. Subretinal fluid was successfully drained in all eyes. The one-operation reattachment rate was 91%, and the final reattachment rate was 98%. Intraoperatively, 15% of eyes required corneal debridement to obtain an adequate view for safe subretinal fluid drainage. Subretinal hemorrhage of no more than one clock hour occurred in 4.2% of eyes. A total of 2.6% of eyes developed an epiretinal membrane postoperatively that required a vitrectomy to improve the visual acuity. The postoperative visual acuity, 0.54 by logMAR (20/69 by Snellen), was significantly better than the preoperative visual acuity, 1.1 logMAR (20/250 by Snellen) (P < 0.0001). The modified external needle drainage technique is a flexible, effective method to drain subretinal fluid in eyes with a rhegmatogenous retinal detachment. The retinal reattachment rate and complication rate compare favorably to alternative subretinal fluid drainage techniques.


Functional and structural pattern of arterial responses in hereditary hypertriglyceridemic and spontaneously hypertensive rats in early stage of experimental hypertension.
It has been shown that endothelium-derived nitric oxide (NO) plays an important role in regulation of vascular tone in the prenatal and early postnatal period. The aim of this paper was to determine the reactivity and accompanying structural changes in thoracic aorta from 4-week-old spontaneously hypertensive rats (SHR) and rats with hereditary hypertriglyceridemia (hHTG) in comparison with age-matched normotensive controls. For functional studies thoracic aorta was excised, cut into rings and mounted in organ baths for measurement of isometric contractile force. For morphological studies cardiovascular system of rats was perfused with glutaraldehyde fixative (at 100 mm Hg) via cannula placed in the left ventricle. Morphological changes of thoracic aorta were measured using light microscopy. Systolic blood pressure (SBP) in SHR (98+/-1 mm Hg) did not significantly differ from that of age-matched control rats (95+/-4 mm Hg), but was slightly increased in hHTG rats (110+/-2 mm Hg, P<0.05). Heart weight/body weight ratio was higher in SHR and hHTG rats than in control group indicating the hypertrophy of the heart in both models of hypertension. Endothelium-dependent relaxation of aorta induced by acetylcholine was preserved in all groups and did not differ from that in control normotensive rats. The maximal isometric contraction of thoracic aorta to noradrenaline (NA) was reduced in hypertensive groups and the concentration-response curves to NA were shifted to the right indicating increased sensitivity of smooth muscle to NA. The values of wall thickness and cross sectional area as well as inner diameter of thoracic aorta in SHR and hHTG rats were significantly decreased in comparison to control groups. Endothelial dysfunction seems to be absent in all young rats before development of hypertension. In conclusion, our observations indicate that in early stage of experimental hypertension NO-dependent relaxation is preserved so that putative impairment of this function provides no significant pathogenic contribution to the onset of hypertension in these two experimental models.


Nuclear reactor accidents--the use of KI as a blocking agent against radioiodine uptake in the thyroid--a review.
This paper is intended for those people who are responsible for the public health and safety in the event of a nuclear reactor accident. The possibilities and consequences of a radioiodine release are examined briefly. The possible side effects of stable iodine, the prognosis for radiation-induced thyroid disease, and the alternative protective measures are put into perspective and assessed for their individual risks and benefits. It is recommended that all appropriate counter-radiation measures be considered in the case of a reactor accident, and that the harmful side effects of the various actions be weighed carefully. Definitive guidelines as to when the use of KI is justified must be decided upon before implementing mass distribution.


[Prevention of coronary atherosclerosis. II: Topography and morphology of "protruding" lesions. Epidemiologic, diagnostic and therapeutic implications].
Topographic and morphologic aspects of coronary atherosclerotic "protruding" lesions were investigated in 119 accidental deaths in males living in Mexico City. Morphology and topography varied according to the arterial trunk studied and age. Left anterior descending and circumflex lesions were almost always confined to the initial 5 cms and if a lesion was present after the 3rd cm there was always a proximal lesion. Right coronary lesions occasionally were found distally even in the absence of proximal lesions. Third decade lesions were usually not calcified. Stenosis if present was rarely multivascular. Some fourth decade lesions were calcified; plurivascular stenosis was present in some cases. Fifth decade lesions show sequential stenotic lesions in the same vessel; calcium and plurivascular stenosis were often observed. However the features observed in the 3rd decade could be observed in the older subjects. Reference is made to lesions which are found in both the left main trunk and the left anterior descending. Epidemiological and clinical applications related with preventive programs are mentioned.


Coronary CT angiography.
Advances in multidetector CT (MDCT) technology with submillimeter slice collimation and high temporal resolution permit contrast-enhanced imaging of coronary arteries and coronary plaque during a single breath hold. Appropriate patient preparation, detailed technical and technological knowledge with regard to recognition of typical imaging artifacts (such as beam hardening or motion artifacts), and the adequate choice of postprocessing techniques to detect stenosis and plaque are prerequisites to achieving diagnostic image quality. A growing number of studies have suggested that 64-slice coronary CT angiography is highly accurate for the exclusion of significant coronary artery stenosis (>50% luminal narrowing), with negative predictive values of 97%-100%, in comparison with invasive selective coronary angiography. In addition, several studies have indicated that MDCT also can detect calcified and noncalcified coronary atherosclerotic plaques, especially in proximal vessel segments, showing a good correlation with intracoronary ultrasound. Studies on clinical utility, cost, and cost-effectiveness are now warranted to demonstrate whether and how this technique can change and improve the current management of patients with suspected or confirmed coronary artery disease.


[Recommendations for basic emergency equipment in the dental office].
Medical emergencies in the dentist's office are a primary concern for dental surgeons. Since each individual therapist is personally responsible for the management of those eventualities, they must be properly trained to handle them. Prevention is still the most effective method for avoiding them, with clinical examination as the prime basis for their management. Some current factors that increase the possibility of office emergencies include: a raise in life expectancy rates, thus increasing the number of patients with chronic diseases; multiple and complex therapeutic regimes; patients under dialysis, immunosuppression, transplants, etc. Apart from proper training of the dental surgeon in managing medical emergencies, he must have adequate equipment for treating such events. This article aims to offer dentists a practical guide to emergency equipment, which must always be at his immediate disposal, as well as some practical guidelines for the use of drugs and some recommendations regarding the organization of such equipment.


Contrasting roles for integrin beta 1 and beta 5 cytoplasmic domains in subcellular localization, cell proliferation, and cell migration.
To carry out a detailed comparison of the roles of integrin beta 1 and beta 5 cytoplasmic domains, we expressed both wild type beta 1 and chimeric beta 1/5 constructs in CHO cells. In the latter, the cytoplasmic domain of beta 1 was replaced with that of beta 5. The human beta 1 and beta 1/5 constructs appeared at similar levels at the cell surface (mostly as alpha 5 beta 1 heterodimers) and contributed equally to CHO cell adhesion to fibronectin. However, beta 1 but not beta 1/5 localized to focal adhesion-like structures when CHO cells were spread on fibronectin. Furthermore, only the beta 1-CHO cells showed increased proliferation in response to fibronectin plus an integrin-activating anti-beta 1 antibody, and showed increased appearance of 32P-labeled protein (p90) that correlated with proliferation. In sharp contrast, the beta 1/5-CHO cells were notably more migratory than beta 1-CHO cells in a transwell haptotactic migration assay. These results indicate that the beta 1 and beta 5 integrin subunit cytoplasmic domains can translate similar adhesive information into highly contrasting subsequent events. Thus, we have established that "inside-out" and "outside-in" integrin signaling pathways are regulated by fundamentally distinct mechanisms. In addition, we suggest that the same properties of the beta 1 cytoplasmic domain that promote recruitment to visible focal adhesion-like structures may also be conductive to cell proliferation. Conversely, the properties of the beta 5 tail that make it less likely to localize into focal adhesion-like structures may contribute to enhanced cell migration.


Health of children in Australian immigration detention centres: An analysis of the quarterly health reports from 2014 to 2017.
This study examines 3 years of child and adolescent health data from Australian onshore and offshore immigration detention centres from 2014 to 2017, quantifying the health presentation data of children and adolescents in Australian immigration detention and comparing rates between onshore and offshore detention. This study utilised the Quarterly Immigration Detention Health Reports over a period of 3 years. To compare onshore and offshore datasets, we calculated the rate of health events per quarter against the estimated quarterly onshore and offshore detention population of children. We ran a series of two-proportion z-tests for each matched quarter to calculate median z and P values for all quarters. These were used as an indicator as to whether the observed differences between onshore and offshore events were statistically significant. The estimated number of children detained per quarter onshore ranged from 700 in 2014 (quarter 3) to 13 in 2016 (quarters 3 and 4); the estimated quarterly population of children in offshore detention ranged from 186 in 2014 (quarter 3) to 42 in 2017 (quarter 2). Children offshore had significantly higher rates of consultations with a mental health nurse (z = -1.96; P = 0.002), psychologist (z = -2.32; P = 0.01) and counsellor (z = -3.41; P < 0.001). As for reasons for presentation to general practitioners and psychiatrists, complaints related to skin (z = -1.97; P = 0.05), respiratory issues (z = -1.96; P = 0.05) and urological issues (z = -2.21; P = 0.03) were significantly higher amongst children detained offshore. Compared to children in the Australian community, children detained both onshore and offshore had greater health needs. Children offshore also presented more frequently with a range of complaints and accessed health services at higher rates than children detained onshore; this adds to growing evidence about the harms of offshore detention and detention more generally.


Biochemical markers of myocardial injury test turnaround time: a College of American Pathologists Q-Probes study of 7020 troponin and 4368 creatine kinase-MB determinations in 159 institutions.
Rapid diagnosis of acute myocardial infarction in patients presenting to emergency departments (EDs) with chest pain may determine the types, and predict the outcomes of, the therapy those patients receive. The amount of time consumed in establishing diagnoses of acute myocardial infarction may depend in part on that consumed in the generation of the blood test results measuring myocardial injury. To determine the normative rates of turnaround time (TAT) for biochemical markers of myocardial injury and to examine hospital and laboratory practices associated with faster TATs. Laboratory personnel in institutions enrolled in the College of American Pathologists Q-Probes Program measured the order-to-report TATs for serum creatine kinase-MB and/or serum troponin (I or T) for patients presenting to their hospital EDs with symptoms of acute myocardial infarction. Laboratory personnel also completed detailed questionnaires characterizing their laboratories' and hospitals' practices related to testing for biochemical markers of myocardial injury. ED physicians completed questionnaires indicating their satisfaction with testing for biochemical markers of myocardial injury in their hospitals. A total of 159 hospitals, predominantly located in the United States, participating in the College of American Pathologists Q-Probes Program. Most (82%) laboratory participants indicated that they believed a reasonable order-to-report TATs for biochemical markers of myocardial injury to be 60 minutes or less. Most (75%) of the 1352 ED physicians who completed satisfaction questionnaires believed that the results of tests measuring myocardial injury should be reported back to them in 45 minutes or less, measured from the time that they ordered those tests. Participants submitted TAT data for 7020 troponin and 4368 creatine kinase-MB determinations. On average, they reported 90% of myocardial injury marker results in slightly more than 90 minutes measured from the time that those tests were ordered. Among the fastest performing 25% of participants (75th percentile and above), median order-to-report troponin and creatine kinase-MB TATs were equal to 50 and 48.3 minutes or less, respectively. Shorter troponin TATs were associated with performing cardiac marker studies in EDs or other peripheral laboratories compared to (1) performing tests in central hospital laboratories, and (2) having cardiac marker specimens obtained by laboratory rather than by nonlaboratory personnel. The TAT expectations of the ED physicians using the results of laboratory tests measuring myocardial injury exceed those of the laboratory personnel providing the results of those tests. The actual TATs of myocardial injury testing meet the expectations of neither the providers of those tests nor the users of those test results. Improving TAT performance will require that the providers and users of laboratory services work together to develop standards that meet the needs of the medical staff and that are reasonably achievable by laboratory personnel.


In vitro exposure to malondialdehyde-acetaldehyde adducted protein inhibits cell proliferation and viability.
Circulating antibodies against malondialdehyde-acetaldehyde (MAA) haptenated proteins are increased significantly in patients with alcohol-induced cirrhosis and hepatitis and are associated with severity of liver damage. Additionally, MAA haptenated proteins are highly immunogenic without the use of adjuvant and have been suggested to induce autoreactive responses. The mechanism of this immunogenicity is currently unknown but may be mediated by cell death in a similar manner as other autoimmune diseases such as systemic lupus erythematosus, mixed connective tissue disease, myositis, and Sjögren's disease. Antigen-presenting cells, lymphocytes, and hepatocytes were exposed to different levels of MAA haptenated hen egg lysozyme and assessed by [3H]thymidine incorporation and (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) conversion for viability. The cells investigated in this study were those that potentially may be involved in the development of autoimmune liver damage; they include antigen-presenting cells, lymphocytes, and cells of the liver itself. Each cell type was found to be sensitive to MAA haptenated protein-induced cell death at levels between 690 microM (10 microg/ml) and 6.9 mM (100 microg/ml), which may be found locally after chronic ethanol consumption. Antigen-presenting cells (macrophage and B cells) were found to be activated at concentrations just under those levels associated with cell death. A dose response to MAA haptenated protein-induced cell death is seen in antigen-presenting cells, lymphocytes, and hepatocytes in vitro. Recent reports have associated both apoptotic and necrotic cell death with the development of autoimmune disease; thus, it is possible that this may be one mechanism by which in vivo immunogenicity is mediated by aldehyde haptenated proteins.


Magnesium- and potassium-sparing effects of amiloride. Review and recent findings.
One of the most common and most serious side-effects of diuretic therapy is an increased urinary loss of potassium. Another, although less well publicized, side-effect of diuretic therapy is excessive loss of urinary magnesium. Loop-blocking diuretics, especially, cause major losses of urinary magnesium, and this is consistent with micropuncture studies in laboratory animals which indicate that the loop of Henle is a major site of magnesium reabsorption. Potassium-sparing diuretics such as amiloride can be administered concomitantly with more potent diuretics to counteract diuretic-induced potassium depletion. Amiloride may also exert magnesium-sparing actions. Our experimental investigations and clinical studies with congestive heart failure patients, providing evidence for both potassium- and magnesium-sparing actions of amiloride, will be discussed. In saline-loaded rats, the magnesium-sparing effect of amiloride was demonstrated when amiloride was administered either alone or in combination with furosemide. Renal clearance studies in rats indicated that the magnesium-sparing effect of amiloride was a direct renal action and not secondary to possible extra-renal actions. A dose-response relationship has been established for the actions of amiloride in reducing fractional excretion of magnesium and potassium during furosemide diuresis in rats. Congestive heart failure patients being treated with furosemide were found to be both potassium and magnesium deficient as indicated by reduced lymphocyte potassium and magnesium. In these patients, amiloride reduced urinary magnesium and potassium, increased plasma magnesium and potassium and also increased lymphocyte magnesium and potassium. The magnesium-sparing actions of amiloride may have important therapeutic implications in that many experimental and clinical studies from our laboratory and from other investigators have shown that magnesium plays an important role in the maintenance and restoration of cellular potassium.


Synthesis of beta-glucans in Prototheca zopfii. Evidence for the existence of a glycoprotein primer.
Membrane preparations from the non-photosynthetic alga Prototheca zopfii incorporate glucose from UDP-[3H]glucose into the trichloroacetic-acid-insoluble fraction and the polysaccharides insoluble in hot alkali. Time course and pulse-chase experiments indicate that the acid-insoluble fraction was a precursor of the alkali-insoluble fraction. Isolation of 3H-labeled membrane or soluble fraction showed that only membrane fractions were able to transfer radioactivity into polysaccharides. Treatment of glucosylated membranes with trypsin or cellulase only partially affect their transfer ability, indicating that the precursor was internalized in vesicles. Analysis of the in vitro synthesized polysaccharides by enzymatic and acid hydrolysis showed that glucose and cellobiose were present as radioactive sugars. Permethylation of the polysaccharide indicates that 80% of the glucose was beta-1,4-bonded with 20% in beta-1,3-linkages. This polysaccharide was found to be identical with the cell-wall beta-glucan obtained in vivo [Rivas, L.A. & Pont Lezica, R. (1978) Planta (Berl.) 165, 348-353].


Effects of chronic exposure to ethanol on the prostaglandin E1 receptor-mediated response and binding in a murine neuroblastoma clone (N1E-115).
With the use of cultured murine neuroblastoma cells (clone N1E-115), the authors studied the effects of chronic ethanol on prostaglandin E, (PGE1)-mediated cyclic AMP formation, adenylate cyclase activity and [3H]PGE1 binding. Whereas acute exposure of these cells to ethanol potentiates the PGE1 response, exposure of cells, for as little as 1 day, to 100 mM ethanol resulted in a diminished responsiveness to PGE1 compared with that in acutely treated cells. This apparent tolerance was well developed by day 4, and, by day 7, treated cells had a diminished response to PGE1 when assayed in the absence of ethanol. To achieve the same level of PGE1-mediated cyclic AMP synthesis as acutely exposed cells, chronically exposed cells required higher concentrations of ethanol. With 7 to 10 days of treatment, there was a modest (10-13%) increase in basal, PGE1- and forskolin-stimulated adenylate cyclase activity in membranous preparations, a 28 to 40% increase in high-affinity [3H]PGE1 binding to membranes with no change in Kd or in the ability of 5'-guanylimidodiphosphate to reduce this binding and a 155% increase in [3H]PGE1 binding to intact cells with no change in Kd. Thus, chronic exposure of N1E-115 cells to ethanol resulted in tolerance to its effects on the PGE1 receptor system, and this tolerance was accompanied by apparently paradoxical changes in PGE1-stimulated cyclic AMP synthesis and [3H]PGE1 binding.


Dysregulation and restoration of homeostatic network plasticity in fragile X syndrome mice.
Chronic activity perturbations in neurons induce homeostatic plasticity through modulation of synaptic strength or other intrinsic properties to maintain the correct physiological range of excitability. Although similar plasticity can also occur at the population level, what molecular mechanisms are involved remain unclear. In the current study, we utilized a multielectrode array (MEA) recording system to evaluate homeostatic neural network activity of primary mouse cortical neuron cultures. We demonstrated that chronic elevation of neuronal activity through the inhibition of GABA(A) receptors elicits synchronization of neural network activity and homeostatic reduction of the amplitude of spontaneous neural network spikes. We subsequently showed that this phenomenon is mediated by the ubiquitination of tumor suppressor p53, which is triggered by murine double minute-2 (Mdm2). Using a mouse model of fragile X syndrome, in which fragile X mental retardation protein (FMRP) is absent (Fmr1 knockout), we found that Mdm2-p53 signaling, network synchronization, and the reduction of network spike amplitude upon chronic activity stimulation were all impaired. Pharmacologically inhibiting p53 with Pifithrin-α or genetically employing p53 heterozygous mice to enforce the inactivation of p53 in Fmr1 knockout cultures restored the synchronization of neural network activity after chronic activity stimulation and partially corrects the homeostatic reduction of neural network spike amplitude. Together, our findings reveal the roles of both Fmr1 and Mdm2-p53 signaling in the homeostatic regulation of neural network activity and provide insight into the deficits of excitability homeostasis seen when Fmr1 is compromised, such as occurs with fragile X syndrome.


Intensive inpatient treatment improves emotion-regulation capacities among adults with severe mental illness.
Impaired capacity for emotion regulation is associated with a broad spectrum of psychiatric disturbances; however, little is known about treatment response in emotion regulation functioning among patients with severe mental illness. This study examined treatment response and the role that experiential avoidance plays in mediating the relationship between attachment anxiety/avoidance and change in emotion regulation. Difficulties in emotion regulation were assessed at admission and at discharge, and rates of improvement and deterioration in emotion regulation were calculated. Attachment anxiety and avoidance were assessed in conjunction with experiential avoidance at baseline in a large cohort (N = 493) of adults admitted to a specialized adult psychiatric hospital. Inpatient treatment was associated with clinically significant improvement in emotion-regulation capacities for 49 percent of patients completing at least four weeks of treatment. Fifty-six percent of patients attained a status of recovery. Greater attachment avoidance and anxiety were related to positive change in emotion regulation at discharge. Experiential avoidance fully mediated the relationship between insecure attachment and change in emotion-regulation capacities. Contrary to expectation, greater attachment insecurity (anxiety and avoidance) as well as greater experiential avoidance predicted improvement in emotion regulation. These counterintuitive findings add to a growing evidence base indicating that severity of psychopathology is associated with greater improvement in hospitalized patients. Results of the mediation analysis suggest that targeting experiential avoidance may be an effective augmentation in the treatment of impaired emotion regulation functioning.


Influenza-induced Tpl2 expression within alveolar epithelial cells is dispensable for host viral control and anti-viral immunity.
Tumor progression locus 2 (Tpl2) is a serine/threonine kinase that regulates the expression of inflammatory mediators in response to Toll-like receptors (TLR) and cytokine receptors. Global ablation of Tpl2 leads to severe disease in response to influenza A virus (IAV) infection, characterized by respiratory distress, and studies in bone marrow chimeric mice implicated Tpl2 in non-hematopoietic cells. Lung epithelial cells are primary targets and replicative niches of influenza viruses; however, the specific regulation of antiviral responses by Tpl2 within lung epithelial cells has not been investigated. Herein, we show that Tpl2 is basally expressed in primary airway epithelial cells and that its expression increases in both type I and type II airway epithelial cells (AECI and AECII) in response to influenza infection. We used Nkx2.1-cre to drive Tpl2 deletion within pulmonary epithelial cells to delineate epithelial cell-specific functions of Tpl2 during influenza infection in mice. Although modest increases in morbidity and mortality were attributed to cre-dependent deletion in lung epithelial cells, no alterations in host cytokine production or lung pathology were observed. In vitro, Tpl2 inhibition within the type I airway epithelial cell line, LET1, as well as genetic ablation in primary airway epithelial cells did not alter cytokine production. Overall, these findings establish that Tpl2-dependent defects in cells other than AECs are primarily responsible for the morbidity and mortality seen in influenza-infected mice with global Tpl2 ablation.