Calcium-activated BKCa channels govern dynamic membrane depolarizations of horizontal cells in rodent retina.
Large conductance, Ca2+ -activated K+ (BKCa ) channels play important roles in mammalian retinal neurons, including photoreceptors, bipolar cells, amacrine cells and ganglion cells, but they have not been identified in horizontal cells. BKCa channel blockers paxilline and iberiotoxin, as well as Ca2+ free solutions and divalent cation Cav channel blockers, eliminate the outwardly rectifying current, while NS1619 enhances it. In symmetrical 150 mm K+ , single channels had a conductance close to 250 pS, within the range of all known BKCa channels. In current clamped horizontal cells, BKCa channels subdue depolarizing membrane potential excursions, reduce the average resting potential and decrease oscillations. The results show that BKCa channel activation puts a ceiling on horizontal cell depolarization and regulates the temporal responsivity of the cells. Large conductance, calcium-activated potassium (BKCa ) channels have numerous roles in neurons including the regulation of membrane excitability, intracellular [Ca2+ ] regulation, and neurotransmitter release. In the retina, they have been identified in photoreceptors, bipolar cells, amacrine cells and ganglion cells, but have not been conclusively identified in mammalian horizontal cells. We found that outward current recorded between -30 and +60 mV is carried primarily in BKCa channels in isolated horizontal cells of rats and mice. Whole-cell outward currents were maximal at +50 mV and declined at membrane potentials positive to this value. This current was eliminated by the selective BKCa channel blocker paxilline (100 nm), iberiotoxin (10 μm), Ca2+ free solutions and divalent cation Cav channel blockers. It was activated by the BKCa channel activator NS1619 (30 μm). Single channel recordings revealed the conductance of the channels to be 244 ± 11 pS (n = 17; symmetrical 150 mm K+ ) with open probability being both voltage- and Ca2+ -dependent. The channels showed fast activation kinetics in response to Ca2+ influx and inactivation gating that could be modified by intracellular protease treatment, which suggests β subunit involvement. Under current clamp, block of BKCa current increased depolarizing membrane potential excursions, raising the average resting potential and producing oscillations. BKCa current activation with NS1619 inhibited oscillations and hyperpolarized the resting potential. These effects underscore the functional role of BKCa current in limiting depolarization of the horizontal cell membrane potential and suggest actions of these channels in regulating the temporal responsivity of the cells.


Transfer Learning with Deep Convolutional Neural Networks for Classifying Cellular Morphological Changes.
The quantification and identification of cellular phenotypes from high-content microscopy images has proven to be very useful for understanding biological activity in response to different drug treatments. The traditional approach has been to use classical image analysis to quantify changes in cell morphology, which requires several nontrivial and independent analysis steps. Recently, convolutional neural networks have emerged as a compelling alternative, offering good predictive performance and the possibility to replace traditional workflows with a single network architecture. In this study, we applied the pretrained deep convolutional neural networks ResNet50, InceptionV3, and InceptionResnetV2 to predict cell mechanisms of action in response to chemical perturbations for two cell profiling datasets from the Broad Bioimage Benchmark Collection. These networks were pretrained on ImageNet, enabling much quicker model training. We obtain higher predictive accuracy than previously reported, between 95% and 97%. The ability to quickly and accurately distinguish between different cell morphologies from a scarce amount of labeled data illustrates the combined benefit of transfer learning and deep convolutional neural networks for interrogating cell-based images.


Sickle cell disease.
Much progress has been made during the past several decades in gaining understanding about the natural history of sickle cell disease and management approaches aimed at treating or even preventing certain disease complications. The characterization of the human genome now offers the opportunity to understand relationships regarding how gene polymorphisms as well as how environmental factors affect the sickle cell disease phenotype, i.e., the individual patient's overall clinical severity as well as their specific organ function. This chapter explores some of these recent advances in knowledge. In Section I, Dr. Michael DeBaun characterizes the problem of silent stroke in sickle cell disease, comparing and contrasting its clinical and neuroimaging features with overt stroke. Combined, these events affect virtually 40% of children with sickle cell anemia. New understanding of risk factors, associated clinical findings, and imaging technologies are impacting substantially on treatment options. The appreciable cognitive dysfunction and other sequelae of silent infarct demand more effective treatments and ultimate prevention. In Section II, Dr. Charles Quinn addresses the conundrum of why some patients with sickle cell disease do well whereas others fare poorly. Some risk factors have been known for years, based upon careful study of hundreds of patients by the Cooperative Study for Sickle Cell Disease and investigators studying the Jamaican newborn cohort. Other prognostic measures have only recently been defined. Dr. Quinn devotes special attention to stroke and chest syndrome as organ-related complications but also describes attempts to measure overall disease severity and to predict survival. Recently, investigators have attempted to predict factors responsible for early mortality in children and following onset of pulmonary hypertension in adults. In Section III, Dr. Martin Steinberg reviews pharmacologic approaches to sickle cell disease and the rationale for their use. In addition to the inhibition of hemoglobin S polymerization, newer targets have been defined during the past one to two decades. These include the erythrocyte membrane, changes in the red cell intracellular content (especially loss of water), endothelial injury, and free radical production. Hydroxyurea treatment attracted the greatest interest, but many uncertainties remain about its long-term benefits and toxicities. Newer "anti-sickling" agents such as decitabine and short-chain fatty acids also receive attention. Prevention of red cell dehydration, "anti-endothelial" therapy, and marshalling the potentially beneficial effects of nitric oxide are other new and exciting approaches.


Mystery pufferfish create elaborate circular nests at mesophotic depths in Australia.
In 2011, the enigma of "mystery circles," small but complex underwater structures first observed by divers from southern Japan in 1995, was solved when a new species of pufferfish, white-spotted pufferfish (Torquigener albomaculosus Matsuura 2014), was identified as the responsible agent. To date these circles have been described only from Japan, where they are formed on a sandy seafloor in water depths less than 30 m. A survey of oil field infrastructure on the North West Shelf of Western Australia in 2018 using a remotely operated vehicle and hybrid autonomous underwater vehicle (HAUV) recorded a high-resolution video and bathymetric data of 21 circular formations with similar features to those described in Japan. The circles display dimensions and morphology like those described from Japan, but were observed in water depths between 129 and 137 m. The HAUV also recorded high-resolution photographs which captured a Torquigener sp. fish in the immediate vicinity of the circles. An additional circle and Torquigener sp. were observed in images collected by baited remote underwater stereo-video in a nearby location in 129 m depth. These circles are the first to be found in Australia. The pufferfish species responsible cannot be identified from images collected. Such a discovery not only generates intrigue and wonder among scientists and the general public but also provides an insight into the reproductive behaviour and evolution of pufferfish globally.


Changes in culture expanded human amniotic epithelial cells: implications for potential therapeutic applications.
Human amniotic epithelial cells (hAEC) isolated from term placenta have stem cell-like properties, differentiate into tissue specific cells and reduce lung and liver inflammation and fibrosis following transplantation into disease models established in mice. These features together with their low immunogenicity and immunosuppressive properties make hAEC an attractive source of cells for potential therapeutic applications. However, generation of large cell numbers required for therapies through serial expansion in xenobiotic-free media may be a limiting factor. We investigated if hAEC could be expanded in xenobiotic-free media and if expansion altered their differentiation capacity, immunophenotype, immunosuppressive properties and production of immunomodulatory factors. Serial expansion in xenobiotic-free media was limited with cumulative cell numbers and population doubling times significantly lower than controls maintained in fetal calf serum. The epithelial morphology of primary hAEC changed into mesenchymal-stromal like cells by passage 4-5 (P4-P5) with down regulation of epithelial markers CK7, CD49f, EpCAM and E-cadherin and elevation of mesenchymal-stromal markers CD44, CD105, CD146 and vimentin. The P5 hAEC expanded in xenobiotic-free medium differentiated into osteocyte and alveolar epithelium-like cells, but not chondrocyte, hepatocyte, α- and β-pancreatic-like cells. Expression of HLA Class IA, Class II and co-stimulatory molecules CD80, CD86 and CD40 remained unaltered. The P5 hAEC suppressed mitogen stimulated T cell proliferation, but were less suppressive compared with primary hAEC at higher splenocyte ratios. Primary and P5 hAEC did not secrete the immunosuppressive factors IL-10 and HGF, whereas TGF-β1 and HLA-G were reduced and IL-6 elevated in P5 hAEC. These findings suggest that primary and expanded hAEC may be suitable for different cellular therapeutic applications.


Functionalization of magnetic nanowires by charged biopolymers.
We report on a facile method for the preparation of biocompatible and bioactive magnetic nanowires. The method consists of the direct deposition of polysaccharides by layer-by-layer (LbL) assembly onto a brush of metallic nanowires obtained by electrodeposition of the metal within the nanopores of an alumina template supported on a silicon wafer. Carboxymethylpullulan (CMP) and chitosan (CHI) multilayers were grown on brushes of Ni nanowires; subsequent grafting of an enzyme was performed by conjugating free amine side groups of chitosan with carboxylic groups of the enzyme. The nanowires are finally released by a gentle ultrasonic treatment. Transmission electron microscopy, electron energy-dispersive loss spectroscopy, and x-ray photoelectron spectroscopy indicate the formation of an homogeneous coating onto the nickel nanowires when one, two, or three CMP/CHI bilayers are deposited. This easy and efficient route to the biochemical functionalization of magnetic nanowires could find widespread use for the preparation of a broad range of nanowires with tailored surface properties.


The Global International Waters Assessment for the Pacific Islands: aspects of transboundary, water shortage, and coastal fisheries issues.
Aspects of transboundary, water shortage, and fisheries issues are discussed in the context of the recently completed Global International Waters Assessment (GIWA) for the Pacific Islands. The region encompasses some 30 million km2 and approximately 12% of the world's ocean space, and features great geographic, demographic and developmental diversity. Global change, especially sea level rise and sea surface temperature increases, is the dominant transboundary concern as it impacts all aspects of life. Water shortage and unsustainable fishing issues are selected for discussion, as they will dominate the region into the foreseeable future, and they are illustrated with examples from Fiji, Kiribati, and Tonga. The environmental impacts are exacerbated by socioeconomic issues such as high population growth rates, urban drift, the breakdown of traditional life styles and the rapid adoption of the cash economy. Policy options that may assist in addressing these issues are proposed.


Analysis of sibling resemblance in anthropometric somatotype components.
We analysed the Heath-Carter somatotype familial resemblance degree in a sample of 1350 siblings (685 males and 665 females) from 634 nuclear families in the province of Biscay (Basque Country, Spain). Sibling correlations were estimated according to maximum likelihood procedures for endomorphy, mesomorphy, ectomorphy and somatotypical attitudinal distance (SADi), after having adjusted for age, sex and the two other components (Model I), and after taking into account several environmental variables shared by all members of each sibship (Model II). All sibling correlations of the somatotype components were significant, tending to be higher in the case of mesomorphy than in the case of endomorphy. Same-sex siblings (BB and SS) displayed significant mesomorphic and ectomorphic resemblances, but in the case of endomorphy, sisters (SS) revealed relatively low correlation compared with brothers (BB). The sex of sibs had a significant influence on correlations of mesomorphy, but not on the other two components. Further control of socio-familial variables such as the father's occupation and education, and the sibship size, resulted in lower mesomorphy correlation coefficients, but had no effect whatsoever on the other two components. SADi correlations were very low and not significant, indicating high degrees of dispersion in the individual somatotypes of the sample.


Immunofluorescent staining with antibodies to factor VIII, fibronectin, and collagenous basement membrane protein in normal human skin and port wine stains.
Specific antibodies directed against three important components of the vessel wall (collagenous basement membrane protein-type IV collagen, fibronectin, and factor VIII) were used to study and compare the distribution of these proteins in normal skin and port wine stains. Collagenous basement membrane protein was localized to the basement membrane and basal lamina zones of blood vessels, appendages, arrector pili muscles, endoneurium and perineurium, and the dermoepidermal junction of both port wine stains and normal skin. Vessels of the port wine stain as well as those of normal skin showed a similar narrow uniform homogeneous line of fluorescence. Granular endothelial staining was seen in the blood vessels of both normal skin and port wine stains. The distribution of fibronectin was that of a low-intensity punctate pattern situated in the subendothelial region of both port wine stain and normal vessels and in the basement membrane zones of hair follicles, endoneurium and perineurium, and the dermoepidermal junction.


Effect of EDTA and Tannic Acid on the Removal of Cd, Ni, Pb and Cu from Artificially Contaminated Soil by Althaea rosea Cavan.
In this study an ornamental plant of Althaea rosea Cavan was investigated for its potential use in the removal of Cd, Ni, Pb and Cu from an artificially contaminated soil. Effect of two different chelating agents on the removal has also been studied by using EDTA (ethylenediaminetetracetic acid) and TA (tannic acid). Both EDTA and TA have led to higher heavy metal concentration in shoots and leaves compared to control plants. However EDTA is generally known as an effective agent in metal solubilisation of soil, in this study, TA was found more effective to induce metal accumulation in Althaea rosea Cavan under the studied conditions. In addition to this, EDTA is toxic to some species and restraining the growth of the plants. The higher BCF (Bio Concentration Factor) and TF (Translocation Factor) values obtained from stems and leaves by the effects of the chemical enhancers (EDTA and TA) show that Althaea rosea Cavan is a hyper accumulator for the studied metals and may be cultivated to clean the contaminated soils.


[Epidemiologic study on dental caries and treatment needs in schoolchildren aged six to twelve years from San Luis Potosi].
To determine the experience, prevalence and severity of dental caries, as well as the Significant Caries Index (SiC) and the treatment needs (TN) in schoolchildren aged six to twelve years. Equally, to estimate the effect of caries in primary dentition on caries in permanent dentition. A cross-sectional study was realized in 3865 schoolchildren from San Luis Potosí City, Mexico. Subjects were clinically evaluated by three standardized examiners (kappa > 0.85). For detection of caries was employed the WHO's criteria for primary (dmft) and permanent (DMFT) dentitions. In the statistical analyzes non-parametric test and logistic regression were used. Mean of age was 8.69 +/- 1.79 and 51.3% were women. In the primary dentition: dmft index was 1.88 +/- 2.34 and the caries prevalence (dmft >0) of 56.8%. In account to severity 22.0% and 5.6% have dmft >3 and dmft >6, respectively. In children of six years, the caries prevalence was 56.0% and the SiC of 5.45. In the permanent dentition: DMFT index was 1.11 +/- 2.03 and the caries prevalence (DMFT >0) of 36.8%. In account to severity 11.8% and 2.8% have DMFT >3 and DMFT >6, respectively. In children of twelve years, the DMFT index caries 4.14 +/- 4.15 and the SiC of 9.15. The TN index was of 88.5% for primary dentition and 93.6% for the permanent. In logistic regression we observed associated to caries prevalence in permanent dentition; presence of caries in primary dentition (OR = 6.37; p < 0.001), female sex (OR = 1.33; p < 0.001) and higher age (OR = 1.69; p < 0.001). Caries prevalence to six years old and DMFT index to twelve years old were highest to values established by WHO. Caries severity in the primary dentition was two times than observed in the permanent dentition. High TN in both dentitions was observed. Presence of caries in primary dentition was a strong risk indicator for to present caries in permanent dentition.


The Importance of Understanding the Stages of COVID-19 in Treatment and Trials.
COVID-19, caused by SARS-CoV-2, continues to be a major health problem since its first description in Wuhan, China, in December 2019. Multiple drugs have been tried to date in the treatment of COVID-19. Critical to treatment of COVID-19 and advancing therapeutics is an appreciation of the multiple stages of this disease and the importance of timing for investigation and use of various agents. We considered articles related to COVID-19 indexed on PubMed published January 1, 2020-November 15, 2020, and considered papers on the medRxiv preprint server. We identified relevant stages of COVID-19 including three periods: pre-exposure, incubation, and detectable viral replication; and five phases: the viral symptom phase, the early inflammatory phase, the secondary infection phase, the multisystem inflammatory phase, and the tail phase. This common terminology should serve as a framework to guide when COVID-19 therapeutics being studied or currently in use is likely to provide benefit rather than harm.


Bigaussian Wavefront Model for Normal and Keratoconic Eyes.
To report bigaussian multivariate wavefront models capable of stochastically generating an unlimited amount of plausible wavefront data for either normal or keratoconic eyes. The models use centroid wavefront data measured previously with an iTrace in 330 healthy right eyes and 122 keratoconic right eyes. These centroids were fitted to an 11th-order Zernike series, followed by principal component analysis to reduce dimensionality and remove correlations. The remaining parameters were then fitted to a sum of two multivariate Gaussian distributions. This fit then forms the core of the stochastic model, which may be used to generate synthetic data. Finally, the agreement between the original and synthetic data was tested using two one-sided t tests. For normal eyes, the first eigenvectors mostly represent pure Zernike polynomials, with a decreasing degree of purity with increasing order. For keratoconic eyes, eigenvector purity was considerably lower than for normal eyes. Depending on the data set, series of 22 to 29 eigenvectors were found sufficient for accurate wavefront reconstruction (i.e., root-mean-square errors below 0.05 μm). These eigenvectors were then used as a base for the stochastic models. In all models and all Zernike coefficients, the mean of the synthetic data was significantly equal to that of the original data (two one-sided t test, P > .05/75), but the variability of the synthetic data is often significantly lower (F test, P < .05/75). This synthetic wavefront model may be safely used in calculations as an alternative to actual measurements should such data not be available.


Stimulation of Alpha1-Adrenergic Receptor Ameliorates Cellular Functions of Multiorgans beyond Vasomotion through PPARδ.
Cells can shift their metabolism between glycolysis and oxidative phosphorylation to enact their cell fate program in response to external signals. Widely distributed α 1-adrenergic receptors (ARs) are physiologically stimulated during exercise, were reported to associate with the activating energetic AMPK pathway, and are expected to have biological effects beyond their hemodynamic effects. To investigate the effects and mechanism of AR stimulation on the physiology of the whole body, various in vitro and in vivo experiments were conducted using the AR agonist midodrine, 2-amino-N-[2-(2,5-dimethoxyphenyl)-2-hydroxy-ethyl]-acetamide. The expression of various biomarkers involved in ATP production was estimated through Western blotting, reverse transcription polymerase chain reaction, oxygen consumption rate, enzyme-linked immunosorbent assay (ELISA), fluorescence staining, and Oil red O staining in several cell lines (skeletal muscle, cardiac muscle, liver, macrophage, vascular endothelial, and adipose cells). In spontaneously hypertensive rats, blood pressure, blood analysis, organ-specific biomarkers, and general biomolecules related to ATP production were measured with Western blot analysis, immunohistochemistry, ELISA, and echocardiography. Pharmacological activation of α 1-adrenergic receptors in C2C12 skeletal muscle cells promoted mitochondrial oxidative phosphorylation and ATP production by increasing the expression of catabolic molecules, including PPARδ, AMPK, and PGC-1α, through cytosolic calcium signaling and increased GLUT4 expression, as seen in exercise. It also activated those energetic molecules and mitochondrial oxidative phosphorylation with cardiomyocytes, endothelial cells, adipocytes, macrophages, and hepatic cells and affected their relevant cell-specific biological functions. All of those effects occurred around 3 h (and peaked 6 h) after midodrine treatment. In spontaneously hypertensive rats, α 1-adrenergic receptor stimulation affected mitochondrial oxidative phosphorylation and ATP production by activating PPARδ, AMPK, and PGC-1α and the relevant biologic functions of multiple organs, suggesting organ crosstalk. The treatment lowered blood pressure, fat and body weight, cholesterol levels, and inflammatory activity; increased ATP content and insulin sensitivity in skeletal muscles; and increased cardiac contractile function without exercise training. These results suggest that the activation of α 1-adrenergic receptor stimulates energetic reprogramming via PPARδ that increases mitochondrial oxidative phosphorylation and has healthy and organ-specific biological effects in multiple organs, including skeletal muscle, beyond its vasomotion effect. In addition, the action mechanism of α 1-adrenergic receptor may be mainly exerted via PPARδ.


Causes and temporal trends of blindness and severe visual impairment in children in schools for the blind in North India.
To describe the causes of severe visual impairment and blindness (SVI/BL) in children in schools for the blind in north India, and explore temporal trends in the major causes. A total of 703 children were examined in 13 blind schools in Delhi. A modified WHO/PBL eye examination record for children with blindness and low vision which included sections on visual acuity, additional non-ocular disabilities, onset of visual loss, the most affected anatomical part of the eye concerning visual impairment, and the aetiological category of the child's disorder based on the timing of insult leading to visual loss was administered in all children. With best correction, 22 (3.1%) were severely visually impaired (visual acuity in the better eye of <6/60) and 628 (89.3%) children were blind (visual acuity in the better eye of <3/60). Anatomical sites of SVI/BL were whole globe in 27.4% children, cornea 21.7%, retina 15.1%, and lens 10.9%. The underlying cause of visual loss was undetermined in 56.5% children (mainly abnormality since birth 42.3% and cataract 8.3%), childhood disorders were responsible in 28.0% (mainly vitamin A deficiency/measles 20.5%), and hereditary factors were identified in 13.4%. Study of temporal trends of SVI/BL by comparing causes in children in three different age groups-5-8 years, 9-12 years, and 13-16 years-suggests that retinal disorders have become more important while childhood onset disorders (particularly vitamin A deficiency) have declined. Almost half of the children suffered from potentially preventable and/or treatable conditions, with vitamin A deficiency/measles and cataract the leading causes. Retinal disorders seem to be increasing in importance while childhood disorders have declined over a period of 10 years.


Risk indicators for recurrent aphthous ulcers among adults in the US.
Recurrent aphthous ulcers (RAU) in the oral cavity are painful, causing substantial morbidity in the US and elsewhere in the world. Despite this, there is a lack of population-based studies representative of the US national adult population to describe the epidemiology, and estimate the true disease burden and association with independent risk factors. Although several studies have investigated the role of various factors in RAU etiology, the epidemiology and etiology of RAU remain unclear. This study aims to establish the prevalence and describe the epidemiology of RAU in adults. Data from the Third National Health and Nutrition Examination Survey (NHANES III) were analyzed in SUDAAN using multivariable logistic regression, modeling RAU occurrences. Overall, for all Americans regardless of age, prevalence of RAU was 1030 per 100,000 people (95% CI 830-1220). The prevalence of RAU among adults was 850 per 100,000 (95% CI 630-1070). The lower vestibule was the most commonly involved site. Multivariable analyses suggested that adjusted odds of RAU were greatest for those 17-29 years of age (adjusted OR 2.7; 95% CI 1.4-5.5), for men (adjusted OR 1.7; 95% CI 0.9-2.8), and for those with low serum insulin levels (OR 2.0; 95% CI 0.9-4.4). Never smokers had greater risk of RAU (OR 9.2, 95% CI 2.8-30.1) compared with those who smoked more than 10 cigarettes per day. This study establishes the prevalence of RAU among adults in the US and demonstrates that whereas cigarette smoking is associated with lesser odds, low insulin levels might be independently associated with greater odds of RAU.


Ferrocene-functionalized polydopamine as a novel redox matrix for H2O2 oxidation.
We describe a simple approach for the synthesis of a ferrocene-derivative of dopamine by reacting ferrocene dicarboxylic acid with the amine functional groups present in dopamine via amide linkage. Polymerization of this derivative was effected at room temperature under mild alkaline conditions (pH ∼ 8.5). The formation of covalent amide linkage between polydopamine (polyDA) and ferrocene (Fc) units was confirmed by FTIR and NMR spectroscopy. Electrochemical characterization of the poly(DA-Fc)-modified glassy carbon electrodes shows a reversible response at Epa = 0.40 V and Epc = 0.28 V pertaining to the ferrocene/ferrocenium redox, in addition to polyDA redox peaks at Epa = 0.63 V and Epc = 0.57 V. Furthermore, the poly(DA-Fc) film displays excellent electrocatalytic oxidation of H2O2 in phosphate buffer solutions (pH ∼ 7.4). A Fenton-type reaction mechanism is proposed to explain the electrocatalytic oxidation, in which oxidation of ferrocene occurs along with the hydroxyl groups in the forward anodic scan, yielding a polymer film with Fc+ and hydroxyl radicals and also a polymer radical, upon reaction with H2O2 gives out oxyhydroxy radicals and ferrocene-polydopamine polymers. During the reverse scan, charge neutralization among the radicals occurs taking the polymer back to its original state. A linear relationship between the anodic current and [H2O2] allows quantitative determination of the latter with a detection limit and sensitivity of 5 μM and 0.7 μA μM-1, respectively. Also, the poly(DA-Fc) film shows no interference from other small organic molecules such as glucose and ascorbic acid.


Dewetting pathways and morphology of unstable thin liquid bilayers.
Three-dimensional long-wave nonlinear analysis of the instabilities engendered by van der Waals forces in a thin (<100 nm) viscous bilayer resting on a rigid substrate is presented. The bilayers are classified based on the macroscopic dewetting behavior of the films, and the three-dimensional morphological evolutions of the films are studied in each case. The dewetting of the bilayers is initiated by one of the two basic modes at the interfaces: in-phase "bending" and out-of-phase "squeezing". We show that the thicknesses, surface energies, and viscosities of the films have significant influence on the mode selection, pathway, of dewetting, and the final morphology of dewetting. Different equilibrium morphologies are obtained by tuning the nature and the strength of the intermolecular forces present at the films, which include: (i) an array of channels and ridges in the lower layer with the upper liquid embedded inside the channels, (ii) islands in the lower layer with the upper layer embedded in the interstitial spaces, and (iii) the lower layer droplets encapsulated by the upper layer.


Effects of Sodium Cromoglycate on Iranian Asthmatic Subjects Without Exposure to any Bronchoconstrictor agent.
Cromolyn sodium, a mast cell stabilizing agent, provides an immediate protective effect against the exercise-induced bronchoconstriction while being used before the exercise. However, cromolyn is ineffective in reversing asthmatic bronchospasm; it is used as a maintenance therapy and has a prophylactic role in chronic asthma. The purpose of this study was to determine the extent of change in baseline lung function tests following a single dose of cromolyn sodium in adult asthmatics. Forty volunteers (33 women and 7 men) with moderate to severe persistent asthma were randomly assigned to receive 20 mg cromolyn, 40 mg cromolyn or cromolyn-placebo. The percent of improvement in lung function parameters was compared among the groups, during 1 h of inhalation. Low dose of cromolyn induced more improvement in most lung function parameters such as forced expiratory flow volume in one second, forced vital capacity and peak expiratory flow compared with other groups. After 15 min, the improvement percentage of baseline forced expiratory flow volume in one second was 3.35 ± 1.5, for sodium cromoglycate-20 mg group compared with 0.98 ± 1.43 and - 0.68 ± 1.2 for sodium cromoglycate-placebo and sodium cromoglycate-40 mg, groups respectively. However, the differences between means were not significant. Furthermore, based on the definition of American Thoracic Society (ATS) for a "significant post-bronchodilator response" developed in a few patients 15 min after the inhalation of 20 mg cromolyn sodium. It is suggested that probably the inhalation of 20 mg of cromolyn sodium could immediately improve the lung function in few adults with asthma.


Caries risk assessment models in caries prediction.
The aim of this research was to assess the efficiency of different multifactor models in caries prediction. Data from the questionnaire and objective examination of 109 examinees was entered into the Cariogram, Previser and Caries-Risk Assessment Tool (CAT) multifactor risk assessment models. Caries risk was assessed with the help of all three models for each patient, classifying them as low, medium or high-risk patients. The development of new caries lesions over a period of three years [Decay Missing Filled Tooth (DMFT) increment = difference between Decay Missing Filled Tooth Surface (DMFTS) index at baseline and follow up], provided for examination of the predictive capacity concerning different multifactor models. The data gathered showed that different multifactor risk assessment models give significantly different results (Friedman test: Chi square = 100.073, p=0.000). Cariogram is the model which identified the majority of examinees as medium risk patients (70%). The other two models were more radical in risk assessment, giving more unfavorable risk -profiles for patients. In only 12% of the patients did the three multifactor models assess the risk in the same way. Previser and CAT gave the same results in 63% of cases - the Wilcoxon test showed that there is no statistically significant difference in caries risk assessment between these two models (Z = -1.805, p=0.071). Evaluation of three different multifactor caries risk assessment models (Cariogram, PreViser and CAT) showed that only the Cariogram can successfully predict new caries development in 12-year-old Bosnian children.


Restructuring wages and benefits to gain a competitive edge.
Healthcare organizations that are able to develop a cost advantage in their wage-and-benefit programs will have a competitive edge as managed care continues to become more pervasive. Organizations can gain an edge by developing pay systems that reward quality, productivity, and cost containment, and by developing benefit systems that provide cost-containment incentives while meeting organizational recruitment and retention objectives.


[A rare and mortal infection agent in patients with hematological malignancies: Saprochaete capitata].
Saprochaete capitata (formerly known as Geotrichum capitatum and Blastoschizomyces capitatus) is a rare invasive fungal agent that may lead to mortal clinical course in patients with hematological malignancies. This agent can be colonized in skin, lungs and intestines, and it can cause major opportunistic infections. Invasive systemic infections due to S.capitata have been reported in immunosuppressed patients. In this report, two patients with invasive S.capitata infections detected during the course of persistent neutropenic fever in acute leukemia, were presented. In both cases empirical caspofungin was added to the treatment, as no response was obtained by board-spectrum antibacterial therapy in neutropenic fever. In the first patient, there were no significant findings except the chronic inflammation observed in the biopsies which was performed for the symptoms of lymphadenitis, myositis, and hepatosplenic candidiasis. While persistent fever was on going, S.capitata was isolated from the blood and catheter cultures. There was no response after catheter removing and the introduction of amphotericin B and voriconazole therapy, therefore allogeneic stem cell transplantation plan for the second time for bone marrow aplasia was taken an earlier time. However, the patient died due to progressive pericardial and pleural effusion and multiorgan failure, although an afebrile process after stem cell transplantation could be obtained. Similarly the second patient had persistent fever despite empirical caspofungin treatment. The additional symptoms of diarrhea, abdominal pain and subileus have indicated an intraabdominal infection. During the follow up, S.capitata was isolated from the blood and catheter cultures. Catheter was removed and amphotericin B was initiated. No response was obtained, and voriconazole was added to treatment. Despite of an afebrile and culture-negative period, the patient died as a result of Acinetobacter sepsis and multiorgan failure. Minimal inhibitory concentration values for both of the Saprochete strains were found as 0.25 µg/ml for amfoterisin B, 1 µg/ml for flukonazol, 0.125 µg/ml for vorikonazol and 0.25 µg/ml for itrakonazol. Virulence model was created by injecting the isolates to the Galleria mellonella larvae, and the life cycle of the larvae were determined. The observation revealed that the infected larvae began to die on the second day and there was no live larvae remained on the eleventh day. In conclusion, S.capitata should be considered as an infection agent with high mortality risk in the neutropenic patients with hematologic malignancies, especially in the presence of persistent fever during the use of caspofungin.


Expansion of Stem Cell-Like CD4+ Memory T Cells during Acute HIV-1 Infection Is Linked to Rapid Disease Progression.
Acute HIV-1 infection is characterized by high viremia and massive depletion of CD4+ T cells throughout all tissue compartments. During this time the latent viral reservoir is established but the dynamics of memory CD4+ T cell subset development, their infectability and influence on disease progression during acute HIV-1 infection has not been carefully described. We therefore investigated the dynamics of CD4+ T cell memory populations in the RV217 (ECHO) cohort during the acute phase of infection. Interestingly, while we found only small changes in central or effector memory compartments, we observed a profound expansion of stem cell-like memory CD4+ T cells (SCM) (2.7-fold; P < 0.0001). Furthermore, we demonstrated that the HIV-1 integration and replication preferentially take place in highly differentiated CD4+ T cells such as transitional memory (TM) and effector memory (EM) CD4+ T cells, while naive and less mature memory cells prove to be more resistant. Despite the relatively low frequency of productively infected SCM, we suggest that their quiescent phenotype, increased susceptibility to HIV-1 integration compared to naive cells and extensive expansion make them one of the key players in establishment and persistence of the HIV-1 reservoir. Moreover, the expansion of SCM in acute HIV-1 infection was a result of Fas upregulation on the surface of naive CD4+ T cells. Interestingly, the upregulation of Fas receptor and expansion of SCM in acute HIV-1 infection was associated with the early viral set point and disease progression (rho = 0.47, P = 0.02, and rho = 0.42, P = 0.041, respectively). Taken together, our data demonstrate an expansion of SCM during early acute HIV-1 infection which is associated with disease outcome.IMPORTANCE Understanding the immunopathology of acute HIV-1 infection will help to develop eradication strategies. We demonstrate here that a CD4+ T cell memory subset expands during acute HIV-1 infection, which is associated with disease progression.


Sealing occlusal caries lesions in adults referred for restorative treatment: 2-3 years of follow-up.
The aim of this study was to assess the possibility to arrest occlusal caries lesions in adults by sealant as well as to assess the presence of radiographic progression, arrest, and regression of the sealed lesions. Seventy-two occlusal caries lesions in 52 adult patients referred to restorative treatment by senior lecturers at School of Dentistry, Copenhagen, Denmark were included. In case the patient had more than one occlusal caries lesion, randomization between sealing and restoration was made; otherwise, the lesion was sealed. In total, 60 resin sealants and 12 composite restorations were made. Follow-up period was 25-38 months (mean = 33 months). Data were analyzed using non-parametric statistics including kappa statistics. After 2-3 years, the dropout rate was 15%; two patients did not show up for control and nine previously sealed lesions were restored by the patients' general practitioners. All 12 restorations and 39 of the remaining 49 sealants were well functioning, seven (14%) sealants were repaired/replaced due to failure, and three (6%) sealed lesions were restored due to caries progression (p > 0.05). The radiographic assessment showed caries progression beneath five (10%) sealants, caries regression beneath one (2%) sealant, and unchanged depth beneath 43 (88%) sealants and all restorations (p > 0.05). The majority of the referred lesions were successfully arrested by sealants, indicating the possibility for extending the criteria for sealing occlusal caries lesions in adults. However, a longer observation period is needed for final conclusion. Extending the criteria of therapeutic sealing of occlusal caries lesions in adults will lead to increased dental health.


[Pain sensitivity threshold in patients with ischemic heart diseases with and without stable angina with episodes of "silent" myocardial ischemia].
A total of 124 coronary patients were examined. Group I comprised 52 patients with stabile angina, group II was 72 patients free from angina. In all patients, 24-hour ECG monitoring was carried out together with veloergometry (VEM), coronaryangiography. Determined in the above patients was also threshold of pain sensitivity (PST) by using the precardiac area electroskin stimulation technique. The frequency of pain-free episodes of myocardial ischemia (PFMI) in subjects with stabile angina has not been found to be different from that in those free from it. There was no difference in depth and duration of depression of ST segment either. No significant differences were found out between PFMI and pain episodes of myocardial ischemia in group I subjects. PST in both groups subjects was not associated with the number of damaged coronary arteries (CA) and presence of the asynergic zones in the left ventricle (LV). In angina-free subjects presenting with the single CA affection, the right coronary artery (RCA) appeared to be affected much more frequently than in those patients with stabile angina. No primary damage to RCA could be seen in associated CA afflictions.


Dietary survey in 40-year-old Edinburgh men.
One hundred and seven healthy 40-yr-old Edinburgh men were asked to participate in a dietary survey involving a 7-d weighed record. Ninety seven successfully completed the record. The results showed that the total energy was made up of 13 per cent protein, 38 per cent fat, 40 per cent carbohydrate and 9 per cent alcohol. A small proportion of the men had exceptionally high alcohol intakes. The mean intake of fat was high, 121 g, it is nevertheless lower than amounts consumed in other areas of Britain. Of the energy from fat, 16 per cent came from saturated-fatty acid, 15 per cent from monounsaturated-fatty acid and 4 per cent from polyunsaturated-fatty acid. Linoleic acid contributed 3 per cent of the mean energy intake. In some mens' diet this essential fatty acid provided under 2 per cent of their total energy consumption.


Endodermal sinus tumor of the mediastinum. A report of seven cases and review of the literature.
Primary mediastinal endodermal sinus tumor is rare, and to date only 49 cases have been described in the English-language literature. Seven new cases are reported. Light microscopic examination showed characteristic features including papillary, reticular, tubular and solid growth patterns, complete or incomplete Schiller-Duval bodies and intracellular or extracellular periodic acid-Schiff-positive material. Immunohistochemical studies showed alpha-1-antitrypsin in seven, alpha-fetoprotein in seven, keratin in six, and carcinoembryonic antigen in four cases. The beta subunit of human chorionic gonadotropin, albumin, fibronectin, and transferrin were not found in any case. Electron microscopic studies performed in four cases showed intracellular and extracellular basement membrane-like material, multiple large multivesicular bodies, desmosomes and microvilli. Mediastinal endodermal sinus tumor can and must be differentiated from poorly differentiated adenocarcinoma metastatic or extending to the mediastinum because of the distinctly different prognoses and therapies. In spite of modern chemotherapy, the prognosis of mediastinal endodermal sinus tumor remains poor. The single most important prognostic indicator is whether the tumor mass can be completely excised before or after chemotherapy.


Second-generation antihistamines for the treatment of chronic idiopathic urticaria.
Chronic idiopathic urticaria (CIU) is a serious disorder that can greatly compromise quality of life. While H1 antihistamines are the accepted first-line treatment for CIU, older generations of these agents (e.g., hydroxyzine, diphenhydramine) are associated with anticholinergic and CNS effects, such as drowsiness and sedation, that can pose risks to patients, especially when driving. Second-generation agents available in the United States (U.S.) (e.g., cetirizine, desloratadine, fexofenadine, levocetirizine, loratadine) has greatly reduced these CNS effects, making them the current treatments of choice in CIU, but their potency and tolerability profiles vary. Differences in duration of in vivo receptor occupancy may affect the potency of H1 antihistamines. Levocetirizine appears to have greater in vivo H1 receptor occupancy compared with later generation H1 antihistamines, which may confer an advantageous efficacy/safety profile. This has been confirmed in a recent head-to-head study showing that levocetirizine was more effective than desloratadine in improving pruritus in CIU patients. Fexofenadine has been shown to have a low occupancy of H1 antihistamine receptors in the brain, which reduces the likelihood of sedation. More studies are required to further assess receptor occupancy and other factors that may differentiate the second-generation H1 antihistamines.


Dynamic contrast enhanced MRI of the axilla in women with breast cancer: comparison with pathology of excised nodes.
Axillary lymph node status is the most important prognostic factor in breast cancer patients and is currently determined by surgical dissection. This study was performed to assess whether dynamic gadopentetate dimeglumine (Gd) enhanced MRI is an accurate method for non-invasive staging of the axilla. 47 women with a new primary breast cancer underwent pre-operative dynamic Gd enhanced MRI of the ipsilateral axilla. Lymph node enhancement was quantitatively analysed using a region of interest method. Enhancement indices and nodal area were compared with histopathology of excised nodes using a receiver operating characteristic (ROC) curve approach. 10 patients had axillary metastases pathologically and all had > or =1 lymph node with an enhancement index of >21% and a nodal area of >0.4 cm(2). 37 patients had negative axillary nodes pathologically. 20 of these had enhancement indices <21% and nodal areas <0.4 cm(2). Using this method, a sensitivity of 100%, a specificity of 56%, a positive predictive value of 38% and a negative predictive value of 100% could be achieved. Using this method of quantitative assessment, dynamic Gd enhanced MRI may be a reliable method of predicting absence of axillary nodal metastases in women with breast cancer, thereby avoiding axillary surgery in women with a negative MRI study.


Comparing the Clinical Features and Trauma Scores of Trauma Patients Aged Under 65 Years with Those of Patients Aged over 65 Years in the Intensive Care Unit: A Retrospective Study for Last Ten Years.
This retrospective study aimed to compare the clinical characteristics and trauma scores of Intensive Care Unit (ICU) trauma patients 65 years and older with the patients under 65 years old. Trauma patients (n=161) who stayed at least 24 hours in ICU were included. Patients younger than 65 years were included into Group 1 (n=109) and patients aged ≥65 years (n=52) were included into Group 2. Patient characteristics and trauma index scores (GCS; APACHE II score, ISS; TRISS and RTS) at ICU admission were calculated. The patients in Group 2 had more comorbid disease compared with Group 1 (61.5%, 6.4%) (p=0.001). The Trauma-related Injury Severity Score score were higher in Group 1 (49.76±33.75) compared with Group 2 (35.38±34.93) (p=0.006). The APACHE II score were higher in Group 2 (20.08±7.60) compared with Group 1 (17.00±6.90) (p=0.007). The need for invasive mechanical ventilation and tracheostomy were more frequent in Group 2 trauma patients compared with those of patients in Group 1 (92.3%, 73.4%; p=0.003; 26.9%, 8.3%; p=0.002; respectively). The need for transfusion of packed red blood cell suspension (PRBC) was more frequent in Group 2 compared with Group 1 (92.3%, 55.0%; respectively) (p=0.001). The mortality rate was found to be higher in Group 2 compared with Group 1 (48.1%, 19.3%; respectively) (p=0.001). The elderly trauma patients have more comorbid disease, higher scores for APACHE II and lower scores for TRISS, more mechanical ventilation and tracheostomy requirements and higher mortality rate compared with young trauma patients.


Physiological dysregulation proceeds and predicts health outcomes similarly in Chinese and Western populations.
A decade ago, we proposed an index of physiological dysregulation based on Mahalanobis distance (DM), that measures how far from the norm an individual biomarker profile is. While extensive validation has been performed, focus was mostly on Western populations with little comparison to developing countries, particularly at a physiological system level. The degree to which the approach would work in other socio-cultural contexts, and the similarity of dysregulation signatures across diverse populations, is still an open question. Using two datasets from China and three from Western countries (USA, United Kingdom, and Italy), we calculated DM, globally and per physiological system. We assessed pairwise correlations among systems, change with age, prediction of mortality and age-related diseases, and sensitivity to interchanging datasets with one another as the reference in DM calculation. Overall, results were comparable across all datasets. Different physiological systems showed distinct dysregulation processes. Association with age was moderate and often nonlinear, similarly for all populations. DM predicted most health outcomes, although differently by physiological system. Using a Chinese population as the reference when calculating DM for Western populations, or vice versa, led to similar associations with health outcomes, with a few exceptions. While small differences were noticeable, they did not systematically emerge between Chinese and Western populations, but rather diffusively across all datasets. These findings suggest that DM presents similar properties, notwithstanding socio-cultural backgrounds, and that it is equally effective in capturing the loss of homeostasis that occurs during aging in diverse industrial human populations.


Up-regulation of MUC2 and IL-1β expression in human colonic epithelial cells by Shigella and its interaction with mucins.
The entire gastrointestinal tract is protected by a mucous layer, which contains complex glycoproteins called mucins. MUC2 is one such mucin that protects the colonic mucosa from invading microbes. The initial interaction between microbes and mucins is an important step for microbial pathogenesis. Hence, it was of interest to investigate the relationship between host (mucin) and pathogen interaction, including Shigella induced expression of MUC2 and IL-1β during shigellosis. The mucin-Shigella interaction was revealed by an in vitro mucin-binding assay. Invasion of Shigella dysenteriae into HT-29 cells was analyzed by Transmission electron microscopy. Shigella induced mucin and IL-1β expression were analyzed by RT-PCR and Immunofluorescence. The clinical isolates of Shigella were found to be virulent by a congo-red binding assay. The in vitro mucin-binding assay revealed both Shigella dysenteriae and Shigella flexneri have binding affinity in the increasing order of: guinea pig small intestinal mucin<guinea pig colonic mucin< Human colonic mucin. Invasion of Shigella dysenteriae into HT-29 cells occurs within 2 hours. Interestingly, in Shigella dysenteriae infected conditions, significant increases in mRNA expression of MUC2 and IL-1β were observed in a time dependent manner. Further, immunofluorescence analysis of MUC2 shows more positive cells in Shigella dysenteriae treated cells than untreated cells. Our study concludes that the Shigella species specifically binds to guinea pig colonic mucin, but not to guinea pig small intestinal mucin. The guinea pig colonic mucin showed a greater binding parameter (R), and more saturable binding, suggesting the presence of a finite number of receptor binding sites in the colonic mucin of the host. In addition, modification of mucins with TFMS and sodium metaperiodate significantly reduced mucin-bacterial binding; suggesting that the mucin-Shigella interaction occurs through carbohydrate epitopes on the mucin backbones. Overproduction of MUC2 may alter adherence and invasion of Shigella dysenteriae into human colonic epithelial cells.


Comparative Transcriptome Analysis Identifies Desmoglein-3 as a Potential Oncogene in Oral Cancer Cells.
The role of desmoglein-3 (DSG3) in oncogenesis is unclear. This study aimed to uncover molecular mechanisms through comparative transcriptome analysis in oral cancer cells, defining potential key genes and associated biological processes related to DSG3 expression. Four mRNA libraries of oral squamous carcinoma H413 cell lines were sequenced, and 599 candidate genes exhibited differential expression between DSG3-overexpressing and matched control lines, with 12 genes highly significantly differentially expressed, including 9 upregulated and 3 downregulated. Genes with known implications in cancer, such as MMP-13, KRT84, OLFM4, GJA1, AMOT and ADAMTS1, were strongly linked to DSG3 overexpression. Gene ontology analysis indicated that the DSG3-associated candidate gene products participate in crucial cellular processes such as junction assembly, focal adhesion, extracellular matrix formation, intermediate filament organisation and keratinocyte differentiation. Validation of RNA-Seq was performed through RT-qPCR, Western blotting and immunofluorescence analyses. Furthermore, using transmission electron microscopy, we meticulously examined desmosome morphology and revealed a slightly immature desmosome structure in DSG3-overexpressing cells compared to controls. No changes in desmosome frequency and diameter were observed between the two conditions. This study underscores intricate and multifaceted alterations associated with DSG3 in oral squamous carcinoma cells, implying a potential oncogenic role of this gene in biological processes that enable cell communication, motility and survival.


Some bitter-sweet reflections on the Ottawa Charter commemoration cake: a personal discourse from an Ottawa rocker.
The Ottawa Charter both gave health promotion a solid framework and health promoters an identity. Yet, health promotion has far from reached its potential in being internalized in public health politics. Advocacy for health is one of the core missions for health promotion and the 25-year celebration of the Ottawa Charter offers a free ride, instead of being a missed opportunity. WHO has not met the expectations in taking advantage of the momentum and outcomes from the long series of global health promotion conferences. The series represents a lifeline for health promotion. Concepts like healthy public policy, supportive environments, social determinants, health and human rights, whole of government, globalization and others have been elaborated and framed in a health promoting context. The downside is that the footprints have not been bold, in particular not internationally. An upside is the development of research and science, underscored by a rapid development of scientific journals, textbooks, academic institutions and posts. A question arising is whether practise and policy making are left behind, since implementation on a grand scale still is lacking? Further and future efforts must be devoted to explore the processes and art of policy making. There is a need for more narratives and more health promoters involving themselves in policy making and politics. Health promotion is as relevant for the twenty-first century as ever. The challenges and opportunities are evident; the increasing global burden of non-communicable diseases, ageing populations, harmful use of alcohol, social determinants and fair societies improved governance and more. Health promotion can add value and WHO can step up its engagement.


A case of unusual substance abuse causing myeloneuropathy.
Case report. Examine an unusual drug related case of myeloneuropathy as well as the pathophysiology of nitrous oxide induced subacute combined degeneration. Major metropolitan teaching hospital - Princess Alexandra Hospital, Queensland, Australia. Review case notes, investigations, relevant medical literature and epidemiological data. A 23-year-old female developed a myeloneuropathy and encephalopathy after an 8-month history of nitrous oxide abuse. Her presentation was complicated by acute renal failure, deep vein thrombosis (DVT) and pulmonary embolism (PE) as well as severe cognitive deficits. After eight months of multidisciplinary rehabilitation the patient is able to walk short distances with mobility aids and is able to manage self cares. However, she still requires a wheelchair for long distances and will have significant residual neurological deficits. The abuse of nitrous oxide has potentially serious outcomes that require discussion of issues related to harm minimisation and health promotion.


Optimized and combined T1 and B1 mapping technique for fast and accurate T1 quantification in contrast-enhanced abdominal MRI.
Fast T(1) mapping techniques are a valuable means of quantitatively assessing the distribution and dynamics of intravenously or orally applied paramagnetic contrast agents (CAs) by noninvasive imaging. In this study a fast T(1) mapping technique based on the variable flip angle (VFA) approach was optimized for accurate T(1) quantification in abdominal contrast-enhanced (CE) MRI. Optimization methods were developed to maximize the signal-to-noise ratio (SNR) and ensure effective RF and gradient spoiling, as well as a steady state, for a defined T(1) range of 100-800 ms and a limited acquisition time. We corrected B(1) field inhomogeneities by performing an additional measurement using an optimized fast B(1) mapping technique. High-precision in vitro and abdominal in vivo T(1) maps were successfully generated at a voxel size of 2.8 x 2.8 x 15 mm(3) and a temporal resolution of 2.3 s per T(1) map on 1.5T and 3T MRI systems. The application of the proposed fast T(1) mapping technique in abdominal CE-MRI enables noninvasive quantification of abdominal tissue perfusion and vascular permeability, and offers the possibility of quantitatively assessing dilution, distribution, and mixing processes of labeled solutions or drugs in the gastrointestinal tract.


Differential DNA mismatch repair underlies mutation rate variation across the human genome.
Cancer genome sequencing has revealed considerable variation in somatic mutation rates across the human genome, with mutation rates elevated in heterochromatic late replicating regions and reduced in early replicating euchromatin. Multiple mechanisms have been suggested to underlie this, but the actual cause is unknown. Here we identify variable DNA mismatch repair (MMR) as the basis of this variation. Analysing ∼17 million single-nucleotide variants from the genomes of 652 tumours, we show that regional autosomal mutation rates at megabase resolution are largely stable across cancer types, with differences related to changes in replication timing and gene expression. However, mutations arising after the inactivation of MMR are no longer enriched in late replicating heterochromatin relative to early replicating euchromatin. Thus, differential DNA repair and not differential mutation supply is the primary cause of the large-scale regional mutation rate variation across the human genome.


Adavosertib plus gemcitabine for platinum-resistant or platinum-refractory recurrent ovarian cancer: a double-blind, randomised, placebo-controlled, phase 2 trial.
The Wee1 (WEE1hu) inhibitor adavosertib and gemcitabine have shown preclinical synergy and promising activity in early phase clinical trials. We aimed to determine the efficacy of this combination in patients with ovarian cancer. In this double-blind, randomised, placebo-controlled, phase 2 trial, women with measurable recurrent platinum-resistant or platinum-refractory high-grade serous ovarian cancer were recruited from 11 academic centres in the USA and Canada. Women were eligible if they were aged 18 years or older, had an Eastern Cooperative Oncology Group performance status of 0-2, a life expectancy of more than 3 months, and normal organ and marrow function. Women with ovarian cancer of non-high-grade serous histology were eligible for enrolment in a non-randomised exploratory cohort. Eligible participants with high-grade serous ovarian cancer were randomly assigned (2:1), using block randomisation (block size of three and six) and no stratification, to receive intravenous gemcitabine (1000 mg/m2 on days 1, 8, and 15) with either oral adavosertib (175 mg) or identical placebo once daily on days 1, 2, 8, 9, 15, and 16, in 28-day cycles until disease progression or unacceptable toxicity. Patients and the team caring for each patient were masked to treatment assignment. The primary endpoint was progression-free survival. The safety and efficacy analysis population comprised all patients who received at least one dose of treatment. The trial is registered with ClinicalTrials.gov, NCT02151292, and is closed to accrual. Between Sept 22, 2014, and May 30, 2018, 124 women were enrolled, of whom 99 had high-grade serous ovarian cancer and were randomly assigned to adavosertib plus gemcitabine (65 [66%]) or placebo plus gemcitabine (34 [34%]). 25 women with non-high-grade serous ovarian cancer were enrolled in the exploratory cohort. After randomisation, five patients with high-grade serous ovarian cancer were found to be ineligible (four in the experimental group and one in the control group) and did not receive treatment. Median age for all treated patients (n=119) was 62 years (IQR 54-67). Progression-free survival was longer with adavosertib plus gemcitabine (median 4·6 months [95% CI 3·6-6·4] with adavosertib plus gemcitabine vs 3·0 months [1·8-3·8] with placebo plus gemcitabine; hazard ratio 0·55 [95% CI 0·35-0·90]; log-rank p=0·015). The most frequent grade 3 or worse adverse events were haematological (neutropenia in 38 [62%] of 61 participants in the adavosertib plus gemcitabine group vs ten [30%] of 33 in the placebo plus gemcitabine group; thrombocytopenia in 19 [31%] of 61 in the adavosertib plus gemcitabine group vs two [6%] of 33 in the placebo plus gemcitabine group). There were no treatment-related deaths; two patients (one in each group in the high-grade serous ovarian cancer cohort) died while on study medication (from sepsis in the experimental group and from disease progression in the control group). The observed clinical efficacy of a Wee1 inhibitor combined with gemcitabine supports ongoing assessment of DNA damage response drugs in high-grade serous ovarian cancer, a TP53-mutated tumour type with high replication stress. This therapeutic approach might be applicable to other tumour types with high replication stress; larger confirmatory studies are required. US National Cancer Institute Cancer Therapy Evaluation Program, Ontario Institute for Cancer Research, US Department of Defense, Princess Margaret Cancer Foundation, and AstraZeneca.


Creativity and health: an anti-humanist reflection.
Many studies suggest that health benefits from engaging with the creative arts, but explanations of the association remain tenuous. This article explores both creativity and health from an anti-humanist perspective and develops a Deleuze-inspired analysis to supply the theoretical framework for creativity and health. In this view, creativity is an active, experimenting flow within a network or assemblage of bodies, things, ideas and institutions, while health is understood as the capacity of a body to affect and be affected by this assemblage. It is consequently unsurprising that there is a relationship between creative activity and health. This analysis is used to explore how creative production and reception can affect health, and to assess the implications for sociology and for arts in health-care practice.


Assessing trauma care systems in low-income and middle-income countries: a systematic review and evidence synthesis mapping the Three Delays framework to injury health system assessments.
The large burden of injuries falls disproportionately on low/middle-income countries (LMICs). Health system interventions improve outcomes in high-income countries. Assessing LMIC trauma systems supports their improvement. Evaluating systems using a Three Delays framework, considering barriers to seeking (Delay 1), reaching (Delay 2) and receiving care (Delay 3), has aided maternal health gains. Rapid assessments allow timely appraisal within resource and logistically constrained settings. We systematically reviewed existing literature on the assessment of LMIC trauma systems, applying the Three Delays framework and rapid assessment principles. We conducted a systematic review and narrative synthesis of articles assessing LMIC trauma systems. We searched seven databases and grey literature for studies and reports published until October 2018. Inclusion criteria were an injury care focus and assessment of at least one defined system aspect. We mapped each study to the Three Delays framework and judged its suitability for rapid assessment. Of 14 677 articles identified, 111 studies and 8 documents were included. Sub-Saharan Africa was the most commonly included region (44.1%). Delay 3, either alone or in combination, was most commonly assessed (79.3%) followed by Delay 2 (46.8%) and Delay 1 (10.8%). Facility assessment was the most common method of assessment (36.0%). Only 2.7% of studies assessed all Three Delays. We judged 62.6% of study methodologies potentially suitable for rapid assessment. Whole health system injury research is needed as facility capacity assessments dominate. Future studies should consider novel or combined methods to study Delays 1 and 2, alongside care processes and outcomes.


The two active sites in human branched-chain alpha-keto acid dehydrogenase operate independently without an obligatory alternating-site mechanism.
A long standing controversy is whether an alternating activesite mechanism occurs during catalysis in thiamine diphosphate (ThDP)-dependent enzymes. We address this question by investigating the ThDP-dependent decarboxylase/dehydrogenase (E1b) component of the mitochondrial branched-chain alpha-keto acid dehydrogenase complex (BCKDC). Our crystal structure reveals that conformations of the two active sites in the human E1b heterotetramer harboring the reaction intermediate are identical. Acidic residues in the core of the E1b heterotetramer, which align with the proton-wire residues proposed to participate in active-site communication in the related pyruvate dehydrogenase from Bacillus stearothermophilus, are mutated. Enzyme kinetic data show that, except in a few cases because of protein misfolding, these alterations are largely without effect on overall activity of BCKDC, ruling out the requirement of a proton-relay mechanism in E1b. BCKDC overall activity is nullified at 50% phosphorylation of E1b, but it is restored to nearly half of the pre-phosphorylation level after dissociation and reconstitution of BCKDC with the same phosphorylated E1b. The results suggest that the abolition of overall activity likely results from the specific geometry of the half-phosphorylated E1b in the BCKDC assembly and not due to a disruption of the alternating active-site mechanism. Finally, we show that a mutant E1b containing only one functional active site exhibits half of the wild-type BCKDC activity, which directly argues against the obligatory communication between active sites. The above results provide evidence that the two active sites in the E1b heterotetramer operate independently during the ThDP-dependent decarboxylation reaction.


Instagram Content Analysis in the Context of the Interpersonal Theory of Suicide.
The Interpersonal Theory of Suicide posits that interpersonal factors, specifically perceived burdensomeness and thwarted belongingness, increase suicidal ideation. Many modern social interactions take place over social media and, as such, examining social media content may offer a novel way to assess interpersonal relationships. In this preregistered study of 386 young adults, we examined the relationship between the amount of interaction with friends- and whether or not individuals interacted with family-related Instagram content and perceived burdensomeness, thwarted belongingness, and suicidal ideation. Instagram content was assessed via self-report and participants' Instagram profiles. Participants who endorsed interacting with family on Instagram demonstrated lower thwarted belongingness and suicidal ideation, but not perceived burdensomeness. Non-preregistered analyses of friend Instagram content demonstrated similar results. Consistent with the Interpersonal Theory, participants' interaction with family and frequency of interacting with friends on Instagram was associated with interpersonal variables and suicidal ideation but not aspects of acquired capability. Our study suggests that social media-based measures of interpersonal relationships are relevant to suicidality.


The myth of the replacement child: parents' stories and practices after perinatal death.
Parents bereaved by perinatal death adapt to their losses in different ways. When bereaved parents give birth to a child or children subsequent to a perinatal death, their constructions of the family necessarily change. The subsequent child is thought to be at risk of psychopathology (the replacement child syndrome) if parents have not sufficiently grieved their losses. This qualitative interview study examines the family stories told by bereaved parents, with particular attention to how parents represent the dead child and subsequent children in the current family structure. We categorized parents' stories as those which suggested that parents replaced the loss by an emphasis on parenting subsequent children, or maintained a connection to the dead child through storytelling and ritual behavior. The two ways in which parents maintained the connection were to preserve the space in the family that the dead child would have inhabited, or to create an on-going relationship with the dead child for themselves and their subsequent children. There seem to be multiple paths to parenting through bereavement. The place of rituals and memorial behavior is also examined.


Permuting the PGF Signature Motif Blocks both Archaeosortase-Dependent C-Terminal Cleavage and Prenyl Lipid Attachment for the Haloferax volcanii S-Layer Glycoprotein.
For years, the S-layer glycoprotein (SLG), the sole component of many archaeal cell walls, was thought to be anchored to the cell surface by a C-terminal transmembrane segment. Recently, however, we demonstrated that the Haloferax volcanii SLG C terminus is removed by an archaeosortase (ArtA), a novel peptidase. SLG, which was previously shown to be lipid modified, contains a C-terminal tripartite structure, including a highly conserved proline-glycine-phenylalanine (PGF) motif. Here, we demonstrate that ArtA does not process an SLG variant where the PGF motif is replaced with a PFG motif (slg(G796F,F797G)). Furthermore, using radiolabeling, we show that SLG lipid modification requires the PGF motif and is ArtA dependent, lending confirmation to the use of a novel C-terminal lipid-mediated protein-anchoring mechanism by prokaryotes. Similar to the case for the ΔartA strain, the growth, cellular morphology, and cell wall of the slg(G796F,F797G) strain, in which modifications of additional H. volcanii ArtA substrates should not be altered, are adversely affected, demonstrating the importance of these posttranslational SLG modifications. Our data suggest that ArtA is either directly or indirectly involved in a novel proteolysis-coupled, covalent lipid-mediated anchoring mechanism. Given that archaeosortase homologs are encoded by a broad range of prokaryotes, it is likely that this anchoring mechanism is widely conserved. Prokaryotic proteins bound to cell surfaces through intercalation, covalent attachment, or protein-protein interactions play critical roles in essential cellular processes. Unfortunately, the molecular mechanisms that anchor proteins to archaeal cell surfaces remain poorly characterized. Here, using the archaeon H. volcanii as a model system, we report the first in vivo studies of a novel protein-anchoring pathway involving lipid modification of a peptidase-processed C terminus. Our findings not only yield important insights into poorly understood aspects of archaeal biology but also have important implications for key bacterial species, including those of the human microbiome. Additionally, insights may facilitate industrial applications, given that photosynthetic cyanobacteria encode uncharacterized homologs of this evolutionarily conserved enzyme, or may spur development of unique drug delivery systems.


Comparison of refractive state and circumferential morphology of retina, choroid, and sclera in chick models of experimentally induced ametropia.
Simultaneous comparisons of the circumferential morphological tissue profiles and final refractions from form-deprivation myopia (FDM), defocus-induced myopia (DIM), and defocus-induced hyperopia (DIH) models of ametropia have been made to test the hypothesis that changes in the thickness profiles of the three coats of the eye, and particularly that of the choroid, can be predicted from the degree of induced refractive error. Hatchling chickens (n = 23) were raised for 2 weeks wearing either a monocular translucent diffuser (FDM, n = 8), monocular -10.00 D lens goggle (DIM, n = 7), monocular +10.00 D lens goggle (DIH, n = 7), or nothing (Norm, n = 1). All animals were refracted using retinoscopy and were then sacrificed, and whole eyes were processed for scanning electron microscopy. Retinal, choroidal, and cartilaginous sclera (CS) thickness measurements were made from photographic collages of the entire circumference of the globe. Of the 23 chickens, complete morphological profile data were available for both eyes of 10 animals (nine treated and one normal). The contralateral fellow eyes (FEyes) of all nine experimental chickens were used as experimental controls as paired comparisons for statistical analyses. Morphological profiles of control and experimental eyes revealed significant systematic regional variations in tissue thickness. This variation was related to nasal or temporal eccentricity with the nasal side generally thinner than the temporal. Retinal, choroidal, and CS tissue from FDM and DIM eyes showed very similar anatomical responses despite significantly different degrees of refractive change. DIH eyes showed significant increases in choroidal thickness but none in retinal or CS thickness. Analysis of fellow control eyes indicated that in both myopia models (FDM and DIM), significant changes in all tissues of the untreated fellow eyes occur whereas only the choroid of the fellow eye was affected in the hyperopic (DIH) model. The morphological similarity observed in the circumferential profiles of the retina, choroid, and cartilaginous sclera of the FDM and DIM eyes despite approximately 20 D difference in final refraction suggests that choroidal thickness is not a good predictor of final refractive error across models. Similarly, the final refractive difference of approximately 20 D between the DIM and the DIH eyes did not receive a major contribution from the final difference in choroidal thickness (with its implied effect on vitreous chamber length).


Over 16% efficiency organic photovoltaic cells enabled by a chlorinated acceptor with increased open-circuit voltages.
Broadening the optical absorption of organic photovoltaic (OPV) materials by enhancing the intramolecular push-pull effect is a general and effective method to improve the power conversion efficiencies of OPV cells. However, in terms of the electron acceptors, the most common molecular design strategy of halogenation usually results in down-shifted molecular energy levels, thereby leading to decreased open-circuit voltages in the devices. Herein, we report a chlorinated non-fullerene acceptor, which exhibits an extended optical absorption and meanwhile displays a higher voltage than its fluorinated counterpart in the devices. This unexpected phenomenon can be ascribed to the reduced non-radiative energy loss (0.206 eV). Due to the simultaneously improved short-circuit current density and open-circuit voltage, a high efficiency of 16.5% is achieved. This study demonstrates that finely tuning the OPV materials to reduce the bandgap-voltage offset has great potential for boosting the efficiency.


Endogenous sex steroids and bone mineral density in older women and men: the Rancho Bernardo Study.
This study examines the associations between endogenous sex steroids and bone mineral density (BMD), using data from a geographically defined cohort in Rancho Bernardo, California. Participants were community-dwelling women and men aged 50-89 years who took part in a study of endogenous sex steroid measurement between 1984-1987 and who had BMD measured in 1988-1991. Those taking corticosteroids or estrogen at the time of sex steroid determination were excluded. The main study outcomes were BMD of the ultradistal radius, midshaft radius, lumbar spine, and total hip by sex steroid level, adjusted for age, body mass index, cigarette smoking, alcohol consumption, leisure exercise, use of thiazides, thyroid hormones, and former estrogen use (women only). At the time of the hormone measurements, the mean age of the 457 women was 72.1 years and that of the 534 men was 68.6 years. A statistically significant positive relation was seen between bioavailable estradiol and BMD at all sites in women and men. Total estradiol was significantly associated with BMD at all sites in women and at all but the ultradistal radius in men. Estrone had a global effect on BMD in women and was not measured in men. Higher bioavailable (but not total) testosterone levels were associated with higher BMD of the ultradistal radius, spine, and hip in men and the ultradistal radius in women. Dehydroepiandrosterone was positively associated with BMD of the midradius, spine, and hip in women and was not associated with BMD at any site in men. Of the sex steroids tested, bioavailable estrogen was most strongly associated with BMD in both women and men. We conclude that endogenous sex steroid levels are significantly related to bone density in older women and men. Individual variation in age-related bone loss may be partially accounted for by alterations in sex steroid levels with aging. Further study to elucidate safe environmental and medical methods to maintain optimal sex steroid levels in old age is needed.


Hospital-to-home transitions for children with medical complexity: part 1, a systematic review of reported outcomes.
Outcome selection to evaluate interventions to support a successful transition from hospital to home of children with medical complexity (CMC) may be difficult due to the variety in available outcomes. To support researchers in outcome selection, this systematic review aimed to summarize and categorize outcomes currently reported in publications evaluating the effectiveness of hospital-to-home transitional care interventions for CMC. We searched the following databases: Medline, Embase, Cochrane library, CINAHL, PsychInfo, and Web of Science for studies published between 1 January 2010 and 15 March 2023. Two reviewers independently screened the articles and extracted the data with a focus on the outcomes. Our research group extensively discussed the outcome list to identify those with similar definitions, wording or meaning. Consensus meetings were organized to discuss disagreements, and to summarize and categorize the data. We identified 50 studies that reported in total 172 outcomes. Consensus was reached on 25 unique outcomes that were assigned to six outcome domains: mortality and survival, physical health, life impact (the impact on functioning, quality of life, delivery of care and personal circumstances), resource use, adverse events, and others. Most frequently studied outcomes reflected life impact and resource use. Apart from the heterogeneity in outcomes, we also found heterogeneity in designs, data sources, and measurement tools used to evaluate the outcomes. Conclusion: This systematic review provides a categorized overview of outcomes that may be used to evaluate interventions to improve hospital-to-home transition for CMC. The results can be used in the development of a core outcome set transitional care for CMC.


From Influenza Virus Infections to Lupus: Synchronous Estrogen Receptor α and RNA Polymerase II Binding Within the Immunoglobulin Heavy Chain Locus.
Males and females respond to pathogens differently and exhibit significantly different frequencies of autoimmune disease. For example, vaccinated adult females control influenza virus better than males, but females suffer systemic lupus erythematosus at a 9:1 frequency compared to males. Numerous explanations have been offered for these sex differences, but most have involved indirect mechanisms by which estrogen, a nuclear hormone, modifies cell barriers or immunity. In search of a direct mechanism, we examined the binding of estrogen receptor α (ERα), a class I nuclear hormone receptor, to the immunoglobulin heavy chain locus. Here, we show that in purified murine B cells, ERα and RNA polymerase II (RNA Pol II) exhibit extraordinarily similar DNA binding patterns. We further demonstrate that ERα preferentially binds adenosine-cytidine (AC)-repeats in the immunoglobulin heavy chain locus when supplemental estrogen is added to purified, lipopolysaccharide-activated B cells. Based on these and previous data, we hypothesize that (i) estrogen guides the binding of ERα and its RNA Pol II partner within the locus, which in turn instructs sterile transcription and class switch recombination (CSR), (ii) ERα binding to AC-repeats modifies the DNA architecture and loops associated with CSR, and (iii) by these mechanisms, estrogen instructs antibody expression. By targeting ERα-DNA interactions in the immunoglobulin heavy chain locus, clinicians may ultimately enhance antibody responses in the context of infectious diseases and reduce antibody responses in the context of allergic or autoimmune reactions.


Arthroscopic Bristow-Latarjet combined with bankart repair restores shoulder stability in patients with glenoid bone loss.
Arthroscopic Bankart repair alone cannot restore shoulder stability in patients with glenoid bone loss involving more than 20% of the glenoid surface. Coracoid transposition to prevent recurrent shoulder dislocation according to Bristow-Latarjet is an efficient but controversial procedure. We determined whether an arthroscopic Bristow-Latarjet procedure with concomitant Bankart repair (1) restored shoulder stability in this selected subgroup of patients, (2) without decreasing mobility, and (3) allowed patients to return to sports at preinjury level. We also evaluated (4) bone block positioning, healing, and arthritis and (5) risk factors for nonunion and coracoid screw pullout. Between July 2007 and August 2010, 79 patients with recurrent anterior instability and bone loss of more than 20% of the glenoid underwent arthroscopic Bristow-Latarjet-Bankart repair; nine patients (11%) were either lost before 2-year followup or had incomplete data, leaving 70 patients available at a mean of 35 months. Postoperative radiographs and CT scans were evaluated for bone block positioning, healing, and arthritis. Any postoperative dislocation or any subjective complaint of occasional to frequent subluxation was considered a failure. Physical examination included ROM in both shoulders to enable comparison and instability signs (apprehension and relocation tests). Rowe and Walch-Duplay scores were obtained at each review. Patients were asked whether they were able to return to sports at the same level and practice forced overhead sports. Potential risk factors for nonhealing were assessed. At latest followup, 69 of 70 (98%) patients had a stable shoulder, external rotation with arm at the side was 9° less than the nonoperated side, and 58 (83%) returned to sports at preinjury level. On latest radiographs, 64 (91%) had no osteoarthritis, and bone block positioning was accurate, with 63 (90%) being below the equator and 65 (93%) flush to the glenoid surface. The coracoid graft healed in 51 (73%), it failed to unite in 14 (20%), and graft osteolysis was seen in five (7%). Bone block nonunion/migration did not compromise shoulder stability but was associated with persistent apprehension and less return to sports. Use of screws that were too short or overangulated, smoking, and age higher than 35 years were risk factors for nonunion. The arthroscopic Bristow-Latarjet procedure combined with Bankart repair for anterior instability with severe glenoid bone loss restored shoulder stability, maintained ROM, allowed return to sports at preinjury level, and had a low likelihood of arthritis. Adequate healing of the transferred coracoid process to the glenoid neck is an important factor for avoiding persistent anterior apprehension. Level IV, therapeutic study. See Instructions for Authors for a complete description of levels of evidence.


Adherence to the food-based Japanese dietary guidelines and prevalence of poor oral health-related quality of life among older Japanese adults in the Kyoto-Kameoka study.
Although better diet quality is inversely related to the risk of geriatric disorders, the association of adherence to dietary guidelines with oral health-related quality of life (OHRQoL) is unclear. We aimed to investigate this association in older Japanese adults. This cross-sectional study included 7984 Japanese participants aged ≥ 65 years from the population-based Kyoto-Kameoka study. Dietary intake was estimated using a validated self-administered FFQ. The scores for adherence to the Japanese Food Guide Spinning Top (range: 0 (worst) to 80 (best)) were calculated. These scores were stratified into quartiles (Qs). Poor OHRQoL was defined as a score ≤ 50 using a 12-item Geriatric Oral Health Assessment Index. The OR and 95 % CI were calculated using multivariable logistic regression and the spline model. Higher adherence score was associated with a lower prevalence of poor OHRQoL (Q1-Q4:36·0 %, 32·1 %, 27·9 % and 25·1 %, respectively). An inverse association was found between the score for adherence to the food-based Japanese dietary guidelines and the OR of poor OHRQoL among all the participants (Q1: reference; Q2: OR, 0·87 (95 % CI: 0·75, 1·00); Q3: OR, 0·77 (95 % CI: 0·66, 0·90); Q4: OR, 0·72 (95 % CI: 0·62, 0·85); Pfor trend < 0·001). These relationships were similar to the results in the spline model. Higher adherence to the food-based Japanese dietary guidelines is inversely associated with the prevalence of poor OHRQoL in older adults. Our results may provide useful insights to improve and maintain oral health.


Ametryn removal by Metarhizium brunneum: Biodegradation pathway proposal and metabolic background revealed.
Ametryn is a representative of a class of s-triazine herbicides absorbed by plant roots and leaves and characterized as a photosynthesis inhibitor. It is still in use in some countries in the farming of pineapples, soybean, corn, cotton, sugar cane or bananas; however, due to the adverse effects of s-triazine herbicides on living organisms use of these pesticides in the European Union has been banned. In the current study, we characterized the biodegradation of ametryn (100 mg L-1) by entomopathogenic fungal cosmopolite Metarhizium brunneum. Ametryn significantly inhibited the growth and glucose uptake in fungal cultures. The concentration of the xenobiotic drops to 87.75 mg L-1 at the end of culturing and the biodegradation process leads to formation of four metabolites: 2-hydroxy atrazine, ethyl hydroxylated ametryn, S-demethylated ametryn and deethylametryn. Inhibited growth is reflected in the metabolomics data, where significant differences in concentrations of L-proline, gamma-aminobutyric acid, L-glutamine, 4-hydroxyproline, L-glutamic acid, ornithine and L-arginine were observed in the presence of the xenobiotic when compared to control cultures. The metabolomics data demonstrated that the presence of ametryn in the fungal culture induced oxidative stress and serious disruptions of the carbon and nitrogen metabolism. Our results provide deeper insights into the microorganism strategy for xenobiotic biodegradation which may result in future enhancements to ametryn removal by the tested strain.


[In vitro sensitivity of Mycobacterium chelonae strains to various antimicrobial agents].
The in vitro susceptibility of 32 Mycobacterium chelonae strains to 10 antimicrobial agents was determined. The sources of the different strains were: clinical samples from patients treated at the Hospital Universitario de Canarias and Hospital del Tórax (General and Chest facilities) and from environmental sources (water supply, sewage, swimming pools and the sea). The susceptibility tests were performed by a broth microdilution method (Mueller-Hinton Broth). The results showed amikacin as the most effective antimicrobial agent against M. chelonae isolates, then ofloxacin and cefoxitin. However no statistical difference was detected among them. The least effective was imipenem, followed by ciprofloxacin and norfloxacin.


Thirty-day mortality in patients undergoing laparotomy for small bowel obstruction.
Small bowel obstruction (SBO) is a common indication for emergency laparotomy. There are currently variations in the timing of surgery for patients with SBO and limited evidence on whether delayed surgery affects outcomes. The aim of this study was to evaluate the impact of time to operation on 30-day mortality in patients requiring emergency laparotomy for SBO. Data were collected from the National Emergency Laparotomy Audit (NELA) on all patients aged 18 years or older who underwent emergency laparotomy for all forms of SBO between December 2013 and November 2015. The primary outcome measure was 30-day mortality, with date of death obtained from the Office for National Statistics. Patients were grouped according to the time from admission to surgery (less than 24 h, 24-72 h and more than 72 h). A multilevel logistic regression model was used to explore the impact of patient factors, primarily delay to surgery, on 30-day mortality. Some 9991 patients underwent emergency laparotomy requiring adhesiolysis or small bowel resection for SBO. The overall mortality rate was 7·2 per cent (722 patients). Within each time group, 30-day mortality rates were significantly worse with increasing age, ASA grade, Portsmouth POSSUM score and level of contamination. Patients undergoing emergency laparotomy more than 72 h after admission had a significantly higher risk-adjusted 30-day mortality rate (odds ratio 1·39, 95 per cent c.i. 1·09 to 1·76). In patients who require an emergency laparotomy with adhesiolysis or resection for SBO, a delay to surgery of more than 72 h is associated with a higher 30-day postoperative mortality rate.


Mucosal-Associated Invariant T Cells Improve Nonalcoholic Fatty Liver Disease Through Regulating Macrophage Polarization.
Mucosal-associated invariant T (MAIT) cells, a novel population of innate-like lymphocytes, have been involved in various inflammatory and autoimmune diseases. However, their role in the development of nonalcoholic fatty liver disease (NAFLD) remains unclear. In this study, we investigated the alterations of phenotype and immunological function of MAIT cells in NAFLD. Analysis of PBMCs in 60 patients with NAFLD and 48 healthy controls (HC) revealed that circulating MAIT cell frequency decreased in NAFLD, especially in the patients with higher serum levels of γ-glutamyl transferase or total triglyceride. Functional alterations of circulating MAIT cells were also detected in NAFLD patients, such as the increased production of IL-4 whereas the decreased production of IFN-γ and TNF-α. Furthermore, elevated expression of CXCR6 was observed in circulating MAIT cells of patients. Meanwhile, we found an increased number of MAIT cells in the livers of NAFLD, and the number was even greater in patients with higher NAFLD activity score. Moreover, activated MAIT cells induced monocytes/macrophages differentiation into M2 phenotype in vitro. Additionally, MAIT cells were enriched and displayed Th2 type cytokines profile in livers of wild type mice fed with methionine and choline deficient diet (MCD). Notably, mice deficient of MAIT cells exhibited more severe hepatic steatosis and inflammation upon MCD, accompanied with more CD11c+ proinflammatory macrophages (M1) and less CD206+ anti-inflammatory macrophages (M2) in livers. Our results indicate that MAIT cells protect against inflammation in NAFLD through producing regulatory cytokines and inducing anti-inflammatory macrophage polarization, which may provide novel therapeutic strategies for NAFLD.


Inhibition of ribosomal peptidyltransferase by chloramphenicol. Kinetic studies.
The mechanism of action of chloramphenicol in inhibiting peptide bond formation has been examined with the aim of discovering whether chloramphenicol brings about conformational changes in the peptidyltransferase domain, its target locus on the ribosome. These conformational changes have been sought as changes in the catalytic rate constant of peptidyltransferase. A detailed kinetic analysis of the inhibition of the puromycin reaction in a system derived from Escherichia coli [Kalpaxis et al. (1986) Eur. J. Biochem. 154, 267-271] has been carried out. There is an initial phase of competitive inhibition (Ki = 0.7 microM) in which the double-reciprocal plots are linear. This phase is observed at concentrations of chloramphenicol up to about 3.0 microM (4.3 Ki). By increasing the concentration of the inhibitor the kinetics change and the inhibition becomes no longer of the competitive type. These results are obtained when the inhibitor is added simultaneously with the substrate (puromycin). Preincubation with the inhibitor before the addition of puromycin gives hyperbolic double-reciprocal plots at inhibitor concentrations around the Ki. After preincubation with the inhibitor at concentrations above the Ki (3-100 Ki) the double-reciprocal plots are linear again and indicate complete, mixed non-competitive inhibition. Analogous behaviour is observed with thiamphenicol (Ki = 0.45 microM) and tevenel (Ki = 1.7 microM). It is proposed that initially chloramphenicol and its two analogs interact with puromycin at a ribosomal locus (peptidyltransferase domain) in a mutually exclusive binding mode (competitive kinetics). Soon after this initial interaction, the antibiotic induces conformational changes to the peptidyltransferase domain so that puromycin is accepted and peptide bonds are still formed but with a lower catalytic rate constant. At this latter state, the ribosome can accept both the inhibitor and the substrate (puromycin) but then, if the concentration of the inhibitor is sufficiently high, peptide bonds are not formed (complete, linear mixed non-competitive inhibition).


Distinct lobes of Limulus ventral photoreceptors. I. Functional and anatomical properties of lobes revealed by removal of glial cells.
Removing the glial cells that encase Limulus ventral photoreceptors allows direct observation of the cell surface. Light microscopy of denuded photoreceptors reveals a subdivision of the cell body into lobes. Often one lobe, but sometimes several, is relatively clear and translucent (the R lobes). The lobe adjacent to the axon (the A lobe) has a textured appearance. Scanning electron microscopy shows that microvilli cover the surface of R lobes and are absent from the surface of A lobes. When a dim spot of light is incident on the R lobe, the probability of evoking a single photon response is two to three orders of magnitude higher than when the same spot is incident on the A lobe. We conclude that the sensitivity of the cell to light is principally a function of the R lobe.


[Characteristics of the glutamate decarboxylase reaction in homogenates of various regions of the rat brain].
The glutamate decarboxylase activity in rough homogenates of cerebellum, cortex and truncal part of the rat brain was studied under different conditions of incubation: in the presence of 25 mM glutamate sodium, 0.4 mM pyridoxal-5'-phosphate and both these components. It is found that the initial glutamate decarboxylase activity in cerebellum homogenates is approximately twice as high as in the cortex and trunk homogenates. Addition of the substrate and cofactor, especially in the combination, stimulates considerably the yield of gamma-aminobutyric acid (GABA) in the glutamate decarboxylase reaction, the most pronounced activation being observed in the truncal homogenates. The glutamate/GABA relation both initial and after the completion of the reaction is the maximal in the cortex and minimal in the truncal part of the brain. The data obtained evidence for the differences in the content of the GABA-producing enzyme rather than for the presence of the specific mechanisms of the enzyme regulation in different brain areas.


Glucose metabolism following endotoxin administration in diabetic rats.
The altered glucose metabolism characteristic of endotoxin-induced shock was studied in diabetic and non-diabetic rats. Endotoxin administration led to an exaggerated hyperglycemia in the diabetic rats. The exaggerated hyperglycemia appeared to be a consequence of an increased hepatic output of glucose, presumably via gluconeogenesis, as evidenced by an exaggerated glucose turnover following endotoxin. Diabetic and non-diabetic rats responded to endotoxin similarly with respect to blood pressure, heart rate, hematocrit, blood pH and plasma lactate. Endotoxin administration did not affect urine flow rate or urinary glucose excretion in either diabetic or non-diabetic rats.