Clinical analysis of patients with pemphigoid-like ophthalmological symptoms.
The aim of this study was a clinical analysis of patients with symptoms typical of pemphigoid. 21 patients were enrolled on the study within the period 1988-2004. The frequency and type of ophthalmological and extraocular symptoms and accompanying diseases were assessed. The analysis also contains immunological findings as well as administered treatment. Ophthalmological changes were detected in 100% of examined patients with pemphigoid. The most frequent were dry eye syndrome and conjunctivitis. Immunohistochemical tests revealed the presence of antibodies in 68.2% of the examined cases. All patients required specific treatment--local, general and surgical. Despite the administered therapy the development of cicatricial changes was observed. Chronic conjunctivitis and keratitis require diagnostics into pemphigoid. Intensive therapy is necessary in patients with ocular cicatricial pemphigoid (OCP) because of high risk of blindness. Delayed diagnostics disable immunohistochemical confirmation of disease.


Postpartum angiographic embolization for vulvovaginal hematoma. A report of two cases.
Puerperal hematomas can become a life-threatening obstetric emergency. Their incidence is 1 or 2/1,000 deliveries. When mainstay methods of suture and packing fail, arterial embolization becomes an excellent alternative to definitive treatment, laparotomy. A 32-year-old woman, para 2-0-2-2, developed extreme rectal and vulvar pain one hour postpartum. A 12 x 10-cm vulvovaginal hematoma was identified, drained, sutured and packed. Bleeding continued, blood products were administered, and selective angiographic embolization of the pudendal and inferior gluteal arteries was successfully performed. A 31-year-old woman, para 1-0-1-1, developed a left vaginal hematoma immediately postpartum. She failed vaginal packing and underwent angiographic arterial embolization successfully. The patient was discharged on the fourth postpartum day. At six weeks postpartum, neither patient had evidence of hematoma formation. In the setting of a puerperal hematoma refractory to conventional first-line therapy, arterial embolization provides a rational, effective alternative for achieving hemostasis with minimal morbidity.


Imprints of function on the folding landscape: functional role for an intermediate in a conserved eukaryotic binding protein.
In the computational characterization of single domain protein folding, the effective free energies of numerous microstates are projected onto few collective degrees of freedom that in turn serve as well-defined reaction coordinates. In this regard, one-dimensional (1D) free energy profiles are widely used mainly for their simplicity. Since folding and functional landscapes are interlinked, how well can these reduced representations capture the structural and dynamic features of functional states while being simultaneously consistent with experimental observables? We investigate this issue by characterizing the folding of the four-helix bundle bovine acyl-CoA binding protein (bACBP), which exhibits complex equilibrium and kinetic behaviours, employing an Ising-like statistical mechanical model and molecular simulations. We show that the features of the 1D free energy profile are sufficient to quantitatively reproduce multiple experimental observations including millisecond chevron-like kinetics and temperature dependence, a microsecond fast phase, barrier heights, unfolded state movements, the intermediate structure and average ϕ-values. Importantly, we find that the structural features of the native-like intermediate (partial disorder in helix 1) are intricately linked to a unique interplay between packing and electrostatics in this domain. By comparison with available experimental data, we propose that this intermediate determines the promiscuous functional behaviour of bACBP that exhibits broad substrate specificity. Our results present evidence to the possibility of employing the statistical mechanical model and the resulting 1D free energy profile to not just understand folding mechanisms but to even extract features of functionally relevant states and their energetic origins.


Dose-Response Relationship of Outdoor Exposure and Myopia Indicators: A Systematic Review and Meta-Analysis of Various Research Methods.
Myopia in children has dramatically increased worldwide. A systematic review and meta-analysis were conducted to evaluate the effects of outdoor light exposure on myopia. According to research data from 13 studies of 15,081 children aged 4-14 at baseline, outdoor light exposure significantly reduced myopia incidence/prevalence (odds ratio [OR] = 0.85, 95% confidence interval [CI]: 0.80-0.91, p < 0.00001; I2 = 90%), spherical equivalent refractive error (SER) by 0.15 D/year (0.09-0.27, p < 0.0001), and axial elongation by 0.08 mm/year (-0.14 to -0.02, p = 0.02). The benefits of outdoor light exposure intervention, according to pooled overall results, included decreases in three myopia indicators: 50% in myopia incidence, 32.9% in SER, and 24.9% in axial elongation for individuals in Asia. Daily outdoor light exposure of more than 120 min was the most effective intervention, and weekly intervention time exhibited a dose-response relationship with all three indicators. Subgroup comparisons revealed that interventional studies report greater benefits from outdoor light exposure compared with cohort and cross-sectional studies, and individuals with myopia in intervention studies experienced slightly greater benefits than individuals without, in terms of SER and axial elongation. Therefore, this study suggests 120 min/day of outdoor light exposure at school.


Measuring diffusion and binding kinetics by contact area FRAP.
The immunological synapse is a stable intercellular structure that specializes in substance and signal transfer from one immune cell to another. Its formation is regulated in part by the diffusion of adhesion and signaling molecules into, and their binding of countermolecules in the contact area. The stability of immunological synapses allows receptor-ligand interactions to approximate chemical equilibrium despite other dynamic aspects. We have developed a mathematical model that describes the coupled reaction-diffusion process in an established immunological synapse. In this study, we extend a previously described contact area fluorescence recovery after photobleaching (FRAP) experiment to test the validity of the model. The receptor binding activity and lateral mobility of fluorescently labeled, lipid-anchored ligands in the bilayer resulted in their accumulation, as revealed by a much higher fluorescence intensity inside the contact area than outside. After complete photobleaching of the synapse, fluorescence recovery requires ligands to dissociate and rebind, and to diffuse in and out of the contact area. Such a FRAP time course consequently provides information on reaction and diffusion, which can be extracted by fitting the model solution to the data. Surprisingly, reverse rates in the two-dimensional contact area were at least 100-fold slower than in three-dimensional solution. As previously reported in immunological synapses, a significant nonrecoverable fraction of fluorescence was observed with one of two systems studied, suggesting some ligands either dissociated or diffused much more slowly compared with other ligands in the same synapse. The combined theory and experiment thus provides a new method for in situ measurements of kinetic rates, diffusion coefficients, and nonrecoverable fractions of interacting molecules in immunological synapses and other stable cell-bilayer junctions.


Extracellular vesicles have variable dose-dependent effects on cultured draining cells in the eye.
The role of extracellular vesicles (EVs) as signal mediators has been described in many biological fields. How many EVs are needed to deliver the desired physiological signal is yet unclear. Using a normal trabecular meshwork (NTM) cell culture exposed to non-pigmented ciliary epithelium (NPCE)-derived EVs, a relevant model for studying the human ocular drainage system, we addressed the EVs dose-response effects on the Wnt signaling. The objective of the study was to investigate the dosing effects of NPCE-derived EVs on TM Wnt signaling. EVs were isolated by PEG 8000 method from NPCE and RPE cells (used as controls) conditioned media. Concentrations were determined by Tunable Resistive Pulse Sensing method. Various exosomes concentration were incubated with TM cells, for the determination of mRNA (β-Catenin, Axin2 and LEF1) and protein (β-Catenin, GSK-3β) expression using real-time quantitative PCR and Western blot, respectively. Exposure of NTM cells for 8 hrs to low EVs concentrations was associated with a significant decreased expression of β-Catenin, GSK-3β, as opposed to exposure to high exosomal concentrations. Pro-MMP9 and MMP9 activities were significantly enhanced in NTM cells treated with high EV concentrations of (X10) as compared to low EV concentrations of either NPCE- or RPE-derived EVs and to untreated control. Our data support the concept that EVs biological effects are concentration-dependent at their target site. Specifically in the present study, we described a general dose-response at the gene and MMPs activity and a different dose-response regarding key canonical Wnt proteins expression.


Photosensitized oxidation of tryptophan: effect on liver and brain tryptophan.
Toxic products are formed when tryptophan is irradiated with light in the presence of photosensitizers such as riboflavin. In order to further investigate this phenomenon, solutions of tryptophan (48 mumol.mL-1) were irradiated with broad spectrum fluorescent light in the presence or absence of riboflavin (0.01 mg.mL-1). Solutions of riboflavin were similarly irradiated; control solutions were the respective solutions not exposed to light. Two-week-old suckling gerbils were then assigned to receiving 7 days of intraperitoneal injections of the light-exposed or non-light-exposed solutions. There were significant differences in the concentrations of tryptophan in serum, liver, and brain; activity of gamma-glutamyl transferase (GGT) as well as liver protein were also significantly different among the groups. Body and liver weights were also significantly different among the groups. In order to identify the photoproducts responsible for these changes, solutions of tryptophan that had been irradiated with light in the presence of riboflavin were then fractionated based on time of elution during high-pressure liquid chromatography and the fractions were then injected into the gerbils as before. GGT responses to one of the fractions was similar to that of the parent compound. Chromatographic studies indicated the presence of numerous photoadduct compounds of tryptophan and riboflavin after exposure to light. Both the presence of riboflavin and the exposure of the solutions to light alter brain concentrations of tryptophan in the developing gerbil indicating differing availability to the brain of this serotonin precursor. Clinical implications of the infusion of amino acids in the presence of photosensitizers and light must be considered.


Frequency discrimination and literacy skills in children with mild to moderate sensorineural hearing loss.
It has been suggested that specific reading disability (SRD) may be attributable to an impaired ability to perceive spectral differences between sounds that leads to a deficit in frequency discrimination and subsequent problems with language and literacy. The objective of the present study was three-fold. We aimed to (a) determine whether children with mild to moderate sensorineural hearing loss were impaired in their ability to discriminate frequency, (b) assess the extent to which any such deficits may be due to an inability to use information derived from phase locking, and (c) examine whether frequency discrimination abilities were predictive of measures of word and nonword reading and nonword repetition. Difference limens for frequency (DLFs) were obtained for 22 children with mild to moderate hearing loss (SNH group) and 22 age-matched controls (CA group) at central frequencies of 1 kHz, where phase-locking information is available, and 6 kHz, where it is not. A battery of standardized tests of language and literacy was also administered. The SNH group exhibited significantly elevated DLFs at both 1 and 6 kHz relative to controls, despite considerable variability of thresholds in both groups. Although no group differences were found for receptive and expressive vocabulary, receptive grammar, and nonword reading, the SNH group performed worse than controls on word reading and nonword repetition, even though word reading scores were age-appropriate. Frequency discrimination abilities were associated with reading and nonword repetition across groups, but these correlations largely disappeared when the two groups were analyzed separately. Together, these results provide evidence for a dissociation between impaired frequency discrimination and relatively "spared" language and literacy in children with mild to moderate sensorineural hearing loss. These results cast doubt on the assertion that a deficit in frequency discrimination necessarily leads to marked deficits in the development of language and literacy.


Beyond the message: advantages of snapshot proteomics with single-cell mass cytometry in solid tumors.
Single-cell technologies that can quantify features of individual cells within a tumor are critical for treatment strategies aiming to target cancer cells while sparing or activating beneficial cells. Given that key players in protein networks are often the primary targets of precision oncology strategies, it is imperative to transcend the nucleic acid message and read cellular actions in human solid tumors. Here, we review the advantages of multiplex, single-cell mass cytometry in tissue and solid tumor investigations. Mass cytometry can quantitatively probe nearly any cellular feature or target. In discussing the ability of mass cytometry to reveal and characterize a broad spectrum of cell types, identify rare cells, and study functional behavior through protein signaling networks in millions of individual cells from a tumor, this review surveys publications of scientific advances in solid tumor biology made with the aid of mass cytometry. Advances discussed include functional identification of rare tumor and tumor-infiltrating immune cells and dissection of cellular mechanisms of immunotherapy in solid tumors and the periphery. The review concludes by highlighting ways to incorporate single-cell mass cytometry in solid tumor precision oncology efforts and rapidly developing cytometry techniques for quantifying cell location and sequenced nucleic acids.


The VIZIER project: preparedness against pathogenic RNA viruses.
Life-threatening RNA viruses emerge regularly, and often in an unpredictable manner. Yet, the very few drugs available against known RNA viruses have sometimes required decades of research for development. Can we generate preparedness for outbreaks of the, as yet, unknown viruses? The VIZIER (VIral enZymes InvolvEd in Replication) (http://www.vizier-europe.org/) project has been set-up to develop the scientific foundations for countering this challenge to society. VIZIER studies the most conserved viral enzymes (that of the replication machinery, or replicases) that constitute attractive targets for drug-design. The aim of VIZIER is to determine as many replicase crystal structures as possible from a carefully selected list of viruses in order to comprehensively cover the diversity of the RNA virus universe, and generate critical knowledge that could be efficiently utilized to jump-start research on any emerging RNA virus. VIZIER is a multidisciplinary project involving (i) bioinformatics to define functional domains, (ii) viral genomics to increase the number of characterized viral genomes and prepare defined targets, (iii) proteomics to express, purify, and characterize targets, (iv) structural biology to solve their crystal structures, and (v) pre-lead discovery to propose active scaffolds of antiviral molecules.


The requirement of pax6 for postnatal eye development: evidence from experimental mouse chimeras.
The small eye mouse mutant (Sey) is caused by a mutation of the Pax6 gene. Previous studies, in which aggregation chimeras were used, have demonstrated that Sey/Sey cells contribute poorly to the neural retina forming small clumps of cells restricted to the inner retina at embryonic day 16.5. In addition, Sey/+ cells are absent from the lens epithelium during this embryonic period and postnatally. This study was conducted to determine the fates of these Sey/Sey and Sey/+ cells with continued development in chimeric mouse eyes. Observations were made on heterozygous and homozygous Sey cells in chimeric eyes from postnatal day (P)0 to P10. In Sey/Sey<-->wild-type (wt) chimeras, all Sey/Sey cells originating from retinal progenitor cells died at perinatal times. The only remaining Sey/Sey cells in the neural retina were associated with blood vessels, including vascular endothelial cells, pericytes, astrocytes, and microglia, which have extraretinal origins. In contrast, Sey/+ cells formed all retinal cell classes. As previously reported, Sey/Sey cells were absent from the lens and corneal epithelium. However, in contrast to previous reports, Sey/+ cells contributed to the lens epithelium as well as corneal tissues, and Sey/Sey cells were absent from the anterior retinal pigment epithelium. All evidence showed that, when Pax6 is absent at the initial stages of the development, Sey/Sey cells that contribute to the neural retina die, even when wild-type cells are available to provide normal environmental cues.


Lung function and breathing pattern in subjects developing high altitude pulmonary edema.
The purpose of the study was to comprehensively evaluate physiologic changes associated with development of high altitude pulmonary edema (HAPE). We tested whether changes in pulmonary function and breathing pattern would herald clinically overt HAPE at an early stage. In 18 mountaineers, spirometry, diffusing capacity, nitrogen washout, nocturnal ventilation and pulse oximetry were recorded at 490 m and during 3 days after rapid ascent to 4559 m. Findings were compared among subjects developing HAPE and those remaining well (controls). In 8 subjects subsequently developing radiographically documented HAPE at 4559 m, median FVC declined to 82% of low altitude baseline while closing volume increased to 164% of baseline (P<0.05, both instances). In 10 controls, FVC decreased slightly (to 93% baseline, P<0.05) but significantly less than in subjects with HAPE and closing volume remained unchanged. Sniff nasal pressure was reduced in both subjects with and without subsequent HAPE. During nights at 4559 m, mean nocturnal oxygen saturation dropped to lower values while minute ventilation, the number of periodic breathing cycles and heart rate were higher (60%; 8.6 L/min; 97 cycles/h; 94 beats/min, respectively) in subjects subsequently developing HAPE than in controls (73%; 5.1 L/min; 48 cycles/h; 79 beats/min; P<0.05 vs. HAPE, all instances). The results comprehensively represent the pattern of physiologic alterations that precede overt HAPE. The changes in lung function are consistent with reduced lung compliance and impaired gas exchange. Pronounced nocturnal hypoxemia, ventilatory control instability and sympathetic stimulation are further signs of subsequent overt HAPE.


Electrocardiogram Derived QRS Duration >120 ms is Associated With Elevated Plasma Homocysteine Levels in a Rural Australian Cross-Sectional Population.
Homocysteine levels in the low to moderate range for cardiovascular risk have been previously associated with left ventricular cardiac hypertrophy (LVH). Electrocardiogram (ECG) derived QRS duration has also been used as an epidemiological screening marker for cardiac hypertrophy risk. QRS duration cut offs have not been previously modeled to assess homocysteine levels in community populations. Our aims are to determine if QRS duration is associated with an elevated homocysteine level in a cross-sectional Australian aging rural population.A retrospective study design utilizing a rural health diabetic screening clinic database containing observational data from the period January 9, 2002 till September 25, 2012. One hundred seventy-eight individuals (>21 years of age) from the database were included in the study. Inclusion criteria included being nondiabetic and having both a QRS duration measure and a matching homocysteine level within the same subject. All participants were from the Albury-Wodonga area, with a mean age of >64 years for both sexes.Mean population homocysteine plasma levels were 10.4 μmol/L (SD = 3.6). The mean QRS duration was 101.8 ms (SD = 17.4). Groups were stratified on the basis of QRS duration (≤120 ms [n = 157] and >120 ms [n = 21]). QRS duration subgroup (≤120 ms vs >120 ms) mean differences across homocysteine levels were 10.1 μmol/L (SD = 3.3) and 12.2 μmol/L (SD = 4.7), respectively (P = 0.016). Other ECG parameters (PQ interval, QTc interval, and QT dispersion) measurements were not significantly associated with differences in plasma homocysteine (P = not significant).We conclude that in community populations homocysteine may be moderately elevated when QRS durations are >120 ms. Small additional increases in homocysteine levels may suggest a risk factor for ECG diagnosis of LVH.


Phenotypic variability of the kyphoscoliotic type of Ehlers-Danlos syndrome (EDS VIA): clinical, molecular and biochemical delineation.
The kyphoscoliotic type of Ehlers-Danlos syndrome (EDS VIA) (OMIM 225400) is a rare inheritable connective tissue disorder characterized by a deficiency of collagen lysyl hydroxylase 1 (LH1; EC 18.104.22.168) due to mutations in PLOD1. Biochemically this results in underhydroxylation of collagen lysyl residues and, hence, an abnormal pattern of lysyl pyridinoline (LP) and hydroxylysyl pyridinoline (HP) crosslinks excreted in the urine. Clinically the disorder is characterized by hypotonia and kyphoscoliosis at birth, joint hypermobility, and skin hyperelasticity and fragility. Severe hypotonia usually leads to delay in gross motor development, whereas cognitive development is reported to be normal. We describe the clinical, biochemical and molecular characterisation, as well as electron microscopy findings of skin, in 15 patients newly diagnosed with this rare type of Ehlers-Danlos syndrome. Age at diagnosis ranged from 5 months to 27 years, with only 1/3 of the patients been diagnosed correctly in the first year of life. A similar disease frequency was found in females and males, however a broad disease severity spectrum (intra- and interfamilial), independent of molecular background or biochemical phenotype, was observed. Kyphoscoliosis, one of the main clinical features was not present at birth in 4 patients. Importantly we also noted the occurrence of vascular rupture antenatally and postnatally, as well as developmental delay in 5 patients. In view of these findings we propose that EDS VIA is a highly variable clinical entity, presenting with a broad clinical spectrum, which may also be associated with cognitive delay and an increased risk for vascular events. Genotype/phenotype association studies and additional molecular investigations in more extended EDS VIA populations will be necessary to further elucidate the cause of the variability of the disease severity.


Vascular regeneration: engineering the stem cell microenvironment.
Vascular diseases are a major threat to human health nowadays. While current treatments can cure some vascular diseases, their beneficial effects are only temporary; vascular regeneration holds the promise of permanent, effective treatments for many vascular diseases. Stem cells and endothelial progenitor cells can differentiate into vascular lineages and therefore have the potential to repair vascular systems. However, engineering appropriate microenvironments that will allow cell maturation and delivery remains the major challenge to the successful implementation of this treatment. This review introduces the cells that are being studied for vascular differentiation and regeneration; we then consider recent approaches to engineering microenvironments, including proper signaling cues and biodegradable scaffolds that will guide the development of these cells into vessels suitable for cell-based vascular therapy.


Coevolution influences the evolution of filial cannibalism, offspring abandonment and parental care.
Understanding evolutionary patterns of parental investment and care has been a long-standing focus in studies of evolutionary and behavioural ecology. Indeed, patterns of investment and care are highly diverse, and fully understanding such diversity has been challenging. Recently, several studies have highlighted the need to consider coevolutionary dynamics in studies of parental care, as parental care is likely to co-occur and co-originate with a range of other traits. Two traits that commonly co-occur with parental care are offspring abandonment (the termination of parental investment prior to full independence in offspring) and filial cannibalism (the consumption of one's offspring). Here, we use a mathematical framework to explore how co-occurrence and coevolution among care, abandonment and cannibalism can influence the life-history conditions under which care is expected to evolve. Our results suggest that in some cases, the evolution of parental care can be inhibited by offspring abandonment and filial cannibalism. In other cases, abandonment and filial cannibalism that benefits parents can promote the evolution of parental care. It is particularly interesting that behaviours that seem so contrary to care-that is, eating or abandoning one's young-can in some cases broaden the conditions under which care can evolve. In general, our findings highlight that considering co-occurrence and coevolutionary dynamics between two or more traits is essential to understanding the evolution of trait diversity.


Ruboxistaurin: LY 333531.
Ruboxistaurin, an orally active protein kinase C beta (PKC beta) inhibitor, is a macrocyclic bisindolylmaleimide compound under development by Eli Lilly with potential as a therapy for diabetic macular oedema and other diabetic angiopathies, including diabetic retinopathy, diabetic peripheral neuropathy and diabetic nephropathy. Ruboxistaurin is awaiting approvals in the US and Europe for the treatment of diabetic retinopathy. Eli Lilly and Alcon entered into a long-term agreement to co-promote ruboxistaurin in the US and Puerto Rico for diabetic retinopathy. The agreement is subject to the US FDA's approval of the agent for this indication. Under the terms of the agreement, Alcon will assume primary responsibility for promotion to eye specialists including retinal specialists and general ophthalmologists, while Eli Lilly will be targeting endocrinologists and physicians. Subject to approval in the US, Eli Lilly will receive milestone and marketing payments from Alcon. Alcon in turn will receive compensation based on product sales. In December 2003, Eli Lilly signed a joint development and co-marketing agreement with Takeda Chemical Industries for ruboxistaurin in the Japanese market. Under the terms of the agreement, Eli Lilly Japan and Takeda will jointly develop ruboxistaurin in Japan, will file an NDA for diabetic peripheral neuropathy and diabetic retinopathy, and subsequently will market the drug in Japan. Ruboxistaurin was submitted for approval in Europe in the second quarter of 2006. The agent is also in phase II studies for the treatment of diabetic maculopathy (macular retinopathy) in Japan. Data from a phase III, 3-year study of ruboxistaurin in patients with moderate to severe diabetic retinopathy showed that ruboxistaurin markedly reduced the risk of sustained vision loss compared with placebo. This multicentre, randomised study, named PKC-DRS2 (Protein Kinase C-Diabetic Retinopathy Study 2), was conducted at 70 clinical sites and involved 685 patients with diabetic retinopathy. The agent is also in a phase II study in the US, Canada and Europe in patients with clinically significant macular oedema. The trial (B7A-MC-MBCU), which will evaluate oral administration of the drug using optical coherence tomography over a period of 18 months, is expected to enrol approximately 220 patients. This randomised, double-blind, placebo-controlled study was initiated in August 2005 and is expected to be completed in March 2008. Previously, results of the PKC-Diabetic Retinopathy Study (PKC-DRS) showed that ruboxistaurin at a dose of 32 mg/day has potential to reduce the risk of moderate vision loss especially in patients with diabetic macular oedema. This phase III, randomised, double-blind, multidose study in 252 patients with type 1 and type 2 diabetes receiving ruboxistaurin or placebo for 3-4 years evaluated the safety of the agent and its effect on progression of diabetic retinopathy, moderate vision loss and sustained moderate vision loss. The study was conducted at Joslin Diabetes Center and at centres in the US, Canada, Denmark, The Netherlands and the UK. In 2004, Eli Lilly presented new analysis of previously reported data for ruboxistaurin in diabetic macular oedema indicating that ruboxistaurin has the potential to decrease the progression of diabetic macular oedema involving the center of the macula. Positive results from the PKC Beta Inhibitor Diabetic Macular Edema (PKC-DMES) trial were reported in 2003. Eli Lilly expected to file for approval of ruboxistaurin for the treatment of diabetic peripheral neuropathy in the US and Europe in 2005. However, no development was reported for this indication. On 15 March 2007, Eli Lilly withdrew its marketing authorisation application for ruboxistaurin for diabetic retinopathy filed with EMEA in May 2006. Its current development status in the EU is unclear at this stage.


Orphans in Three Sahelian Countries: Exploratory Analyses from Census Data.
Important investments in Africa have reduced slightly the levels child mortality but life expectancy still very low. The number of children without surviving biological parents is increasing and orphans are becoming an important social problem. Because Sahelian societies are mostly patriarchal, becoming fatherless or motherless will have different effects on the well being of the child. This paper examines the levels and trends of the survival status of the parents and then, living arrangements of orphans. We describe characteristics of these children with a special focus on education and economic activities. The paper uses the censuses from Chad, Niger and Senegal made available by the African Census Analysis Project (ACAP) held at University of Pennsylvania. These countries collected information on survival status of each biological parent to estimate adult mortality but the potential of this information for research on child well-being is rarely exploited.


Use of posttreatment imaging and biomarkers in survivors of early-stage breast cancer: Inappropriate surveillance or necessary care?
Advanced imaging and serum biomarkers are commonly used for surveillance in patients with early-stage breast cancer, despite recommendations against this practice. Incentives to perform such low-value testing may be less prominent in integrated health care delivery systems. The purpose of the current study was to evaluate and compare the use of these services within 2 integrated systems: Kaiser Permanente (KP) and Intermountain Healthcare (IH). The authors also sought to distinguish the indication for testing: diagnostic purposes or routine surveillance. Patients with American Joint Committee on Cancer stage 0 to II breast cancer diagnosed between 2009 and 2010 were identified and the use of imaging and biomarker tests over an 18-month period were quantified, starting at 1 year after diagnosis. Chart abstraction was performed on a random sample of patients who received testing to identify the indication for testing. Multivariate regression was used to explore associations with the use of nonrecommended care. A total of 6585 patients were identified; 22% had stage 0 disease, 44% had stage I disease, and 34% had stage II disease. Overall, 24% of patients received at least 1 imaging test (25% at KP vs 22% at IH; P = .009) and 28% of patients received at least 1 biomarker (36% at KP vs 13% at IH; P<.001). Chart abstraction revealed that 84% of imaging tests were performed to evaluate symptoms or signs. Virtually all biomarkers were ordered for routine surveillance. Stage of disease, medical center that provided the services, and provider experience were found to be significantly associated with the use of biomarkers. Advanced imaging was most often performed for appropriate indications, but biomarkers were used for nonrecommended surveillance. Distinguishing between inappropriate use for surveillance and appropriate diagnostic testing is essential when evaluating adherence to recommendations.


Contradiction and Complacency Shape Attitudes towards the Toll of Roads on Wildlife.
Most people in the world now live in cities. Urbanisation simultaneously isolates people from nature and contributes to biodiversity decline. As cities expand, suburban development and the road infrastructure to support them widens their impact on wildlife. Even so, urban communities, especially those on the peri-urban fringe, endeavour to support biodiversity through wildlife friendly gardens, green spaces and corridors, and conservation estates. On one hand, many who live on city fringes do so because they enjoy proximity to nature, however, the ever increasing intrusion of roads leads to conflict with wildlife. Trauma (usually fatal) to wildlife and (usually emotional and financial) to people ensues. Exposure to this trauma, therefore, should inform attitudes towards wildlife vehicle collisions (WVC) and be linked to willingness to reduce risk of further WVC. While there is good anecdotal evidence for this response, competing priorities and better understanding of the likelihood of human injury or fatalities, as opposed to wildlife fatalities, may confound this trend. In this paper we sought to explore this relationship with a quantitative study of driver behaviour and attitudes to WVC from a cohort of residents and visitors who drive through a peri-urban reserve (Royal National Park) on the outskirts of Sydney, Australia. We distributed a self-reporting questionnaire and received responses from 105 local residents and 51 visitors to small townships accessed by roads through the national park. We sought the respondents' exposure to WVC, their evasive actions in an impending WVC, their attitudes to wildlife fatalities, their strategies to reduce the risk of WVC, and their willingness to adopt new ameliorative measures. The results were partitioned by driver demographics and residency. Residents were generally well informed about mitigation strategies but exposure led to a decrease in viewing WVC as very serious. In addition, despite most respondents stating they routinely drive slower when collision risk is high (at dusk and dawn), our assessment of driving trends via traffic speeds suggested this sentiment was not generally adhered to. Thus we unveil some of the complexities in tackling driver's willingness to act on reducing risk of WVC, particularly when risk of human trauma is low.


Ligand structure, conformational dynamics, and excited-state electron delocalization for control of photoinduced electron transfer rates in synthetic donor-bridge-acceptor systems.
Synthesis, ground-, and excited-state properties are reported for two new electron donor-bridge-acceptor (D-B-A) molecules and two new photophysical model complexes. The D-B-A molecules are [Ru(bpy)2(bpy-phi-MV)](PF6)4 (3) and [Ru(tmb)2(bpy-phi-MV)](PF6)4 (4), where bpy is 2,2'-bipyridine, tmb is 4,4',5,5'-tetramethyl-2,2'-bipyridine, MV is methyl viologen, and phi is a phenylene spacer. Their model complexes are [Ru(bpy)2(p-tol-bpy)](PF6)2 (1) and [Ru(tmb)2(p-tol-bpy)](PF6)2 (2), where p-tolyl-bpy is 4-(p-tolyl)-2,2'-bipyridine. Photophysical characterization of 1 and 2 indicates that 2.17 eV and 2.12 eV are stored in their respective (3)MLCT (metal-to-ligand charge transfer) excited state. These values along with electrochemical measurements show that photoinduced electron transfer (D*-B-A-->D (+)-B-A(-)) is favorable in 3 and 4 with DeltaG degrees(ET)=-0.52 eV and -0.62 eV, respectively. The driving force for the reverse process (D(+)-B-A(-) --> D-B-A) is also reported: DeltaG degrees(BET)=-1.7 eV for 3 and -1.5 eV for 4. Transient absorption (TA) spectra for 3 and 4 in 298 K acetonitrile provide evidence that reduced methyl viologen is observable at 50 ps following excitation. Detailed TA kinetics confirm this, and the data are fit to a model to determine both forward (k(ET)) and back (k(BET)) electron transfer rate constants: k(ET)=2.6 x 10(10) s(-1) for 3 and 2.8 x 10(10) s(-1) for 4; k(BET)=0.62 x 10(10) s(-1) for 3 and 1.37 x 10(10) s(-1) for 4. The similar rate constants k ET for 3 and 4 despite a 100 meV driving force (DeltaG degrees(ET)) increase suggests that forward electron transfer in these molecules in room temperature acetonitrile is nearly barrierless as predicted by the Marcus theory. The reduction in electron transfer reorganization energy necessary for this barrierless reactivity is attributed to excited-state electron delocalization in the (3)MLCT excited states of 3 and 4, an effect that is made possible by excited-state conformational changes in the aryl-substituted ligands of these complexes.


Undiagnosed pulmonary tuberculosis deaths in rural South Africa.
This research explores death from pulmonary tuberculosis (PTB) using a verbal autopsy (VA) tool within the established Agincourt Health and Demographic Surveillance System site in South Africa's rural northeast. Previous work on active case finding in the area highlighted a modest burden of undiagnosed PTB in the community. This VA research confirms the existence of undiagnosed PTB deaths, with 13 (46%) of 28 PTB deaths among the permanent adult population (n = 38,251) going undetected by the health service. There was a median duration of coughing in the community of 16 weeks among these undiagnosed PTB deaths. As most undiagnosed cases present to the health service at some point during their illness, intervention strategies to support early diagnosis at this level can only be re-emphasised by this work.


Identification of risk factors in atypical chronic myeloid leukemia.
In the WHO classification atypical chronic myeloid leukemia (CML) has been considered as a new distinct clinical entity included in the category of mixed myeloproliferative/myelodysplastic disorders. Little is known about this uncommon disease, whose incidence is about of one-two cases every 100 cases of Ph-positive CML. We analyzed our series of 55 patients diagnosed as having aCML, with the aim of identifying clinical factors of possible prognostic value on survival and acute transformation.


Comparative study of the self-aggregation of rhodamine 6G in the presence of poly(sodium 4-styrenesulfonate), poly(N-phenylmaleimide-co-acrylic acid), poly(styrene-alt-maleic acid), and poly(sodium acrylate).
The interaction between rhodamine 6G and different polyelectrolytes is analyzed. Structural aspects differentiate these polyelectrolytes, such as the presence of aromatic groups and the number and localization of their respective charges, which may be directly attached to the aromatic groups or to the polymeric main chain. In the case of poly(sodium acrylate), which does not bear aromatic groups, the polyelectrolyte induces cooperative self-stacking between the dyes which is highly sensitive to the ionic strength, due to the predominance of long-range electrostatic interactions between the polymer and the dye. In the case of poly(sodium 4-styrenesulfonate), whose charge is directly attached to the aromatic groups, a high dispersant ability of the dyes is found and the interaction is less dependent on the ionic strength, due to the predominance of short-range aromatic-aromatic interactions between the dye and the polymer. Among the two polyelectrolytes studied for which the polymeric charge is directly attached to the main chain, and separated from the aromatic group, poly(styrene-alt-maleic acid) shows a lower dependence of the interaction on the ionic strength than poly(N-phenylmaleimide-co-acrylic acid) at a comonomer composition of 1:2, due to a higher linear aromatic density and a lower linear charge density, indicating the importance of hydrophobic forces. Both copolymers exhibit a high ability to induce cooperative self-aggregation of the dye.


RETRACTED: Direct imaging discovery of a Jovian exoplanet within a triple-star system.
Direct imaging allows for the detection and characterization of exoplanets via their thermal emission. We report the discovery via imaging of a young Jovian planet in a triple-star system and characterize its atmospheric properties through near-infrared spectroscopy. The semimajor axis of the planet is closer relative to that of its hierarchical triple-star system than for any known exoplanet within a stellar binary or triple, making HD 131399 dynamically unlike any other known system. The location of HD 131399Ab on a wide orbit in a triple system demonstrates that massive planets may be found on long and possibly unstable orbits in multistar systems. HD 131399Ab is one of the lowest mass (4 ± 1 Jupiter masses) and coldest (850 ± 50 kelvin) exoplanets to have been directly imaged.


A 1-night operant learning task without food-restriction differentiates among mouse strains in an automated home-cage environment.
Individuals are able to change their behavior based on its consequences, a process involving instrumental learning. Studying instrumental learning in mice can provide new insights in this elementary aspect of cognition. Conventional appetitive operant learning tasks that facilitate the study of this form of learning in mice, as well as more complex operant paradigms, require labor-intensive handling and food deprivation to motivate the animals. Here, we describe a 1-night operant learning protocol that exploits the advantages of automated home-cage testing and circumvents the interfering effects of food restriction. The task builds on behavior that is part of the spontaneous exploratory repertoire during the days before the task. We compared the behavior of C57BL/6J, BALB/cJ and DBA/2J mice and found various differences in behavior during this task, but no differences in learning curves. BALB/cJ mice showed the largest instrumental learning response, providing a superior dynamic range and statistical power to study instrumental learning by using this protocol. Insights gained with this home-cage-based learning protocol without food restriction will be valuable for the development of other, more complex, cognitive tasks in automated home-cages.


RANTES is a chemotactic and activating factor for human eosinophils.
RANTES is a member of the 8-kDa cytokine family that has been shown to possess chemotactic activity for monocytes and CD4 T cells. In this study, we investigated whether RANTES could affect eosinophil chemotaxis and function. Peripheral blood eosinophils from blood donors were isolated on Percoll gradients to > 98% purity and then used for chemotaxis, flow cytometry, eosinophil cationic protein release assay, and survival assay. We found that RANTES is chemotactic for eosinophils at 10(-9) to 10(-8) M concentrations. RANTES elicited 65% of the chemotactic response to 10(-7) M platelet-activating factor in all experiments. The mechanism of chemotaxis was investigated by studying the expression of adhesion molecules on eosinophils by flow cytometry. We found that RANTES up-regulated the expression of CD11b/CD18 on eosinophils in a dose-dependent manner. In another set of experiments, purified eosinophils incubated with various concentrations of RANTES released eosinophil cationic protein as measured by a RIA. We also investigated the effect of RANTES on eosinophil density. Leukocytes were incubated in the presence or absence of RANTES, and the distribution of eosinophils on discontinuous Percoll gradients was then examined. We found that eosinophils became hypodense (< 1.085) when incubated in RANTES. However, unlike IL-3, RANTES did not affect the survival of eosinophils in a 4-day culture system. Thus, we established that RANTES is a chemotactic and activating factor for eosinophils.


Quantitative and selective polymerase chain reaction analysis of highly similar human alpha-class glutathione transferases.
Alpha-class glutathione transferases (GSTs) found expressed in human tissues constitute a family of four homologous enzymes with contrasting enzyme activities. In particular, GST A3-3 has been shown to contribute to the biosynthesis of steroid hormones in human cells and is selectively expressed in steroidogenic tissues. The more ubiquitous GST A1-1, GST A2-2, and GST A4-4 appear to be primarily involved in detoxification processes and are expressed at higher levels than GST A3-3. We are interested in studying the cell and tissue expression of the GST A3-3 gene, yet the existence of highly expressed sequence-similar homologs and of several splice variants is a serious challenge for the specific detection of unique transcript species. We found that published polymerase chain reaction (PCR) primers for GST A3-3 lack the specificity required for reliable quantitative analysis. Therefore, we designed quantitative PCR (qPCR) primers with greatly increased discrimination power for the human GSTA3 full-length transcript. The improved primers allow accurate discrimination between GST A3-3 and the other alpha-class GSTs and so are of great value to studies of the expression of the GSTA3 gene. The novel primers were used to quantify GSTA3 transcripts in human embryonic liver and steroidogenic cell lines.


Development and implementation of EPID-based quality assurance tests for the small animal radiation research platform (SARRP).
Although small animal image-guided radiotherapy (SA-IGRT) systems are used increasingly in preclinical research, tools for performing routine quality assurance (QA) have not been optimized and are not readily available. Robust, efficient, and reliable QA tools are needed to ensure the accuracy and reproducibility of SA-IGRT systems. Several investigators have reported custom-made phantoms and protocols for SA-IGRT systems QA. These are typically time and resource intensive and are therefore not well suited to the preclinical radiotherapy environment, in which physics support is limited and routine QA is performed by technical staff. We investigated the use of the inbuilt electronic portal imaging device (EPID) to develop and validate routine QA tests and procedures. In this work, we focus on the Xstrahl Small Animal Radiation Research Platform (SARRP) EPID. However, the methodology and tests developed here are applicable to any SA-IGRT system that incorporates an EPID. We performed a comprehensive characterization of the dosimetric properties of the camera-based EPID at kilovoltage energies over a 11-month period, including detector warm-up time, radiation dose history effect, stability and short- and long-term reproducibility, gantry angle dependency, output factor, and linearity of the EPID response. We developed a test to measure the constancy of beam quality in terms of half-value layer and tube peak potential using the EPID. We verified the SARRP daily output and beam profile constancy using the imager. We investigated the use of the imager to monitor beam-targeting accuracy at various gantry and couch angles. The EPID response was stable and reproducible, exhibiting maximum variations of ≤0.3% and ≤1.9% for short and long terms, respectively. The detector showed no dependence on response at different gantry angles, with a maximum variation ≤0.5%. We found close agreement in output factor measurement between the portal imager and reference dosimeters, with maximum differences ≤3% for ionization chamber and ≤1.7% for Gafchromic EBT3 dosimetry film, respectively. We have shown that the EPID response is linear with tube current (mA) for the entire range of tube kilovoltage peak. Notably, a close relationship was seen between the detector response vs mA slope, and the kilovoltage peak, allowing an independent verification of kilovoltage peak stability based solely on EPID response. In addition to dosimetry tests, according to the beam-targeting measurement using portal images, maximum displacement of the central axis of the x-ray beam (due to sag) was 0.76 ± 0.09 mm at gantry 135°/couch 0° and 0.89 ± 0.06 mm at gantry 0°/couch -135°. We performed the first comprehensive analysis on the dosimetric properties of an EPID operating at kilovoltage x-ray energies. We characterized the detector performance over a 11-month period. Our results indicate that the imager is a stable and convenient tool for SARRP routine QA tests. We then developed EPID-based tests to perform routine SA-IGRT systems QA tasks, such as verifying constancy of beam quality, energy, output, and profile measurements, relative output factors, and beam targeting.


Recurrent nocturnal hypoglycaemia as a cause of morning fatigue in treated Addison's disease--favourable response to dietary management: a case report.
Addison's disease, or primary adrenal insufficiency, is often associated with reduced well-being and fatigue despite use of currently recommended adrenal hormone replacement. Hypoglycaemia is a known manifestation of glucocorticoid deficiency, but is generally considered rare in adults and not relevant to troubling ongoing symptoms in patients with Addison's disease. A 43 year old woman with a three year history of Addison's disease complained of severe morning fatigue and headaches, despite standard glucocorticoid replacement therapy in the form of thrice daily hydrocortisone and mineralocorticoid replacement with fludrocortisone. Alternative glucocorticoid replacement regimens and the addition of dehydroepiandrosterone replacement therapy had no effect. Nocturnal hypoglycaemia was suspected and a 4-day continuous glucose monitor system (CGMS) revealed hypoglycaemia (interstitial glucose < 2.2 mmol/L) between 0200-0400 h on 3 of 4 days. The patient was counselled to take an evening snack designed to ensure slow absorption of ingested carbohydrates. Nocturnal hypoglycaemia was then absent on follow up CGMS assessment. The patient noted a marked symptomatic improvement in morning symptoms, but with persistent fatigue during the day. Currently, the best strategy for control of non-specific symptoms in treated Addison's disease is unknown, but it may be that investigation for hypoglycaemia and treatment, where necessary, could assist some sufferers to achieve improved wellbeing. A systematic study of this phenomenon in Addison's disease is required.


The clinical course of idiopathic dilated cardiomyopathy. A population-based study.
To describe the prognosis of individuals with idiopathic dilated cardiomyopathy in a population-based sample and to compare this with the prognosis of patients in a previous referral center case series of idiopathic dilated cardiomyopathy. Cohort study. Population-based in Olmsted County, Minnesota. Forty residents of Olmsted County, Minnesota with idiopathic dilated cardiomyopathy initially diagnosed between 1975 and 1984 who were followed through 1 July 1989 and 104 patients from a Mayo Clinic referral case series from 1960 to 1973. Survival for the population-based cohort at 1 year and 5 years. Survival at 1 year differed dramatically between the population-based cohort and the referral case series at 1 year (95% compared with 69%, respectively) and at 5 years (80% compared with 36%, respectively) (P less than 0.001). Long-term survival for the population-based cohort was nonetheless impaired when compared with an age- and sex-matched cohort, that is, the 1980 Minnesota white population (8-year survival: observed, 58% compared with expected, 83%; P less than 0.001). Among community patients, older age (adjusted Cox model hazard ratio for 10-year increase in age, 1.59; 95% CI, 1.08 to 2.35) and lower left ventricular ejection fraction (adjusted hazard ratio for 10% decrease, 1.90; CI, 1.04 to 3.50) were independently associated with impaired survival. These population-based data challenge the clinical perception of the clinical course of idiopathic dilated cardiomyopathy based on referral practice prognostic studies and suggest that the clinical course of this condition may be more favorable than previously recognized.


Differential effects of hypothalamic IGF-I on gonadotropin releasing hormone neuronal activation during steroid-induced LH surges in young and middle-aged female rats.
Interactions between brain IGF-I receptors and estrogen receptors regulate female reproductive physiology and behavior. The present study investigated potential mechanisms by which IGF-I receptors in the neuroendocrine hypothalamus regulate GnRH neuronal activation and LH release in young and middle-aged female rats under estradiol (E2) positive feedback conditions. We infused vehicle, IGF-I, or JB-1, a selective antagonist of IGF-I receptors, into the third ventricle of ovariectomized female rats primed with E2 and progesterone or vehicle. In young females, blockade of IGF-I receptors attenuated the steroid hormone-induced LH surge, reduced the percent of GnRH neurons expressing c-fos on the day of the LH surge, and decreased the total number of neurons expressing c-fos in the preoptic area. Middle-aged females had fewer GnRH neurons expressing c-fos during the LH surge than young females, and the LH surge amplitude was attenuated. Infusion of an IGF-I dose previously shown to increase LH surge amplitude did not increase the percent of GnRH neurons expressing c-fos in middle-aged females. Brain IGF-I receptor blockade did not modify E2 induction of progestin receptor-immunoreactive neurons in the preoptic area, arcuate, or ventromedial hypothalamus of young rats. These findings indicate that brain IGF-I receptors are required for E2 activation of GnRH neurons in young rats and for robust GnRH release from axon terminals in middle-aged females. IGF-I likely exerts its effects by actions on E2-sensitive neurons that are upstream of GnRH neurons and terminals.


Fetal growth restriction has long-term effects on postnatal lung structure in sheep.
We have previously shown that fetal growth restriction (FGR) during late gestation in sheep affects lung development in the near-term fetus and at 8 wk after birth. In the present study, our aim was to determine the effects of FGR on the structure of the lungs at 2 y after birth; our hypothesis was that changes observed at 8 wk after birth would persist until maturity. FGR was induced in sheep by umbilicoplacental embolization, which was maintained from 120 d until delivery at term (approximately 147 d); birth weights of FGR lambs were 41% lower than in controls. At 2 y after birth, body and lung weights were not different, but there were 28% fewer alveoli and alveoli were significantly larger than in controls; hence there was a 10% reduction in the internal surface area relative to lung volume in FGR sheep compared with controls. The lungs of FGR sheep, compared with controls, had thicker interalveolar septa as a result of increased extracellular matrix deposition; the alveolar blood-air barrier was also thicker, largely because of an 82% increase in basement membrane thickness. These changes are qualitatively similar to those observed at 8 wk. Our data show that structural alterations in the lungs induced by FGR that were apparent at 8 wk were still evident at 2 y after birth, indicating that FGR may result in permanent changes in the structure of the lungs of the offspring and may affect respiratory health and lung aging later in life.


Outstanding Reviewers for Chemical Science in 2022.
We would like to take this opportunity to highlight the Outstanding Reviewers for Chemical Science in 2022, as selected by the editorial team for their significant contribution to the journal.


Negative selection by an endogenous retrovirus promotes a higher-avidity CD4+ T cell response to retroviral infection.
Effective T cell responses can decisively influence the outcome of retroviral infection. However, what constitutes protective T cell responses or determines the ability of the host to mount such responses is incompletely understood. Here we studied the requirements for development and induction of CD4+ T cells that were essential for immunity to Friend virus (FV) infection of mice, according to their TCR avidity for an FV-derived epitope. We showed that a self peptide, encoded by an endogenous retrovirus, negatively selected a significant fraction of polyclonal FV-specific CD4+ T cells and diminished the response to FV infection. Surprisingly, however, CD4+ T cell-mediated antiviral activity was fully preserved. Detailed repertoire analysis revealed that clones with low avidity for FV-derived peptides were more cross-reactive with self peptides and were consequently preferentially deleted. Negative selection of low-avidity FV-reactive CD4+ T cells was responsible for the dominance of high-avidity clones in the response to FV infection, suggesting that protection against the primary infecting virus was mediated exclusively by high-avidity CD4+ T cells. Thus, although negative selection reduced the size and cross-reactivity of the available FV-reactive naïve CD4+ T cell repertoire, it increased the overall avidity of the repertoire that responded to infection. These findings demonstrate that self proteins expressed by replication-defective endogenous retroviruses can heavily influence the formation of the TCR repertoire reactive with exogenous retroviruses and determine the avidity of the response to retroviral infection. Given the overabundance of endogenous retroviruses in the human genome, these findings also suggest that endogenous retroviral proteins, presented by products of highly polymorphic HLA alleles, may shape the human TCR repertoire that reacts with exogenous retroviruses or other infecting pathogens, leading to interindividual heterogeneity.


Drosophila SMN2 minigene reporter model identifies moxifloxacin as a candidate therapy for SMA.
Spinal muscular atrophy is a rare and fatal neuromuscular disorder caused by the loss of alpha motor neurons. The affected individuals have mutated the ubiquitously expressed SMN1 gene resulting in the loss or reduction in the survival motor neuron (SMN) protein levels. However, an almost identical paralog exists in humans: SMN2. Pharmacological activation of SMN2 exon 7 inclusion by small molecules or modified antisense oligonucleotides is a valid approach to treat SMA. Here we describe an in vivo SMN2 minigene reporter system in Drosophila motor neurons that serves as a cost-effective, feasible, and stringent primary screening model for identifying chemicals capable of crossing the conserved Drosophila blood-brain barrier and modulating exon 7 inclusion. The model was used for the screening of 1100 drugs from the Prestwick Chemical Library, resulting in 2.45% hit rate. The most promising candidate drugs were validated in patient-derived fibroblasts where they proved to increase SMN protein levels. Among them, moxifloxacin modulated SMN2 splicing by promoting exon 7 inclusion. The recovery of SMN protein levels was confirmed by increased colocalization of nuclear gems with Cajal Bodies. Thus, a Drosophila-based drug screen allowed the discovery of an FDA-approved small molecule with the potential to become a novel therapy for SMA.


Variation and comparative effectiveness of patent ductus arteriosus pharmacotherapy in extremely low birth weight infants.
Patent ductus arteriosus (PDA) occurs in 70% of extremely low birth weight (ELBW, birth weight <1000 g) infants. Approximately 34% of ELBW infants with a PDA have spontaneous closure. Failure of the ductus arteriosus to close has been associated with multiple morbidities. To examine variability over time and across hospitals in early therapeutic (2-7 day) use of indomethacin (INDO) vs ibuprofen (IBU) for PDA treatment in outborn ELBW infants and examine the outcomes and side effects of both pharmacological agents in this population. Data were extracted from the Pediatric Health Information System. ELBW infants born between January 1, 2007 and December 31, 2010 and admitted on day of life 0 were eligible for inclusion. 732 infants had a PDA diagnosis and met inclusion criteria. We explored the variability in PDA pharmacotherapy over time and across hospitals. We compared outcomes of both agents for in-hospital mortality, need for surgical ligation, intraventricular hemorrhage, necrotizing enterocolitis, bronchopulmonary dysplasia, periventricular leukomalacia, renal failure, and persistent pulmonary hypertension. Statistical methods included chi square and multivariable regression analysis. Instrumental variable analysis was used to control for selection bias and omitted variables. There was large variability in PDA pharmacotherapy over time and across hospitals. INDO use declined as IBU use grew from 12.8 to 38.9%. There was no difference in hospital or NICU characteristics between high and low IBU using NICUs. Renal failure was more common in infants receiving INDO compared to IBU. We noted large variability in PDA pharmacotherapy. Renal failure was more common with INDO use. Until further studies to compare the long-term effects of both drugs, our data support IBU as the preferred medication for PDA pharmacotherapy in ELBW infants.


Efficient sampling of atomic configurational spaces.
We describe a method to explore the configurational phase space of chemical systems. It is based on the nested sampling algorithm recently proposed by Skilling (AIP Conf. Proc. 2004, 395; J. Bayesian Anal. 2006, 1, 833) and allows us to explore the entire potential energy surface (PES) efficiently in an unbiased way. The algorithm has two parameters which directly control the trade-off between the resolution with which the space is explored and the computational cost. We demonstrate the use of nested sampling on Lennard-Jones (LJ) clusters. Nested sampling provides a straightforward approximation for the partition function; thus, evaluating expectation values of arbitrary smooth operators at arbitrary temperatures becomes a simple postprocessing step. Access to absolute free energies allows us to determine the temperature-density phase diagram for LJ cluster stability. Even for relatively small clusters, the efficiency gain over parallel tempering in calculating the heat capacity is an order of magnitude or more. Furthermore, by analyzing the topology of the resulting samples, we are able to visualize the PES in a new and illuminating way. We identify a discretely valued order parameter with basins and suprabasins of the PES, allowing a straightforward and unambiguous definition of macroscopic states of an atomistic system and the evaluation of the associated free energies.


Time since first sexual intercourse and the risk of cervical cancer.
Young age at first sexual intercourse (AFI) is an important risk factor for cervical cancer, but no simple statistical model of its influence has been established. We investigated the relationship between risk of cervical carcinoma and time since first intercourse using data on monogamous women (5,074 cases and 16,137 controls) from the International Collaboration of Epidemiological Studies of Cervical Cancer. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated from pooled data on 20 studies using conditional logistic regression. The OR for invasive cervical carcinoma is approximately proportional to the square of time since first intercourse (exponent 1.95, 95% CI: 1.76-2.15) up to age 45. First cervical infection with human papillomavirus (HPV) often occurs soon after first sexual intercourse, so early AFI is a reasonable proxy for early age at first exposure to HPV. In addition, age-specific incidence rates of cervical cancer in unscreened populations remain fairly constant above age 45. Cervical cancer thus resembles other cancers caused by strong early-stage carcinogens, with incidence rates proportional to a power of time since first exposure and also resembles cancers of the breast and other hormone-dependent epithelia, where a similar flattening of age-specific incidence rates is seen at the time menopausal changes start. Taken together, these observations suggest that HPV vaccination may prevent the majority of cervical cancers by delaying HPV infection without necessarily providing lifetime protection against HPV.


Prognostic significance of peritumoral lymphatic vessel density and vascular endothelial growth factor receptor 3 in invasive squamous cell cervical cancer.
Cervical cancer is known to metastasize primarily by the lymphatic system. Dissemination through lymphatic vessels represents an early step in regional tumor progression, and the presence of lymphatic metastasis is associated with a poor prognosis. In patients who have undergone a radical hysterectomy, lymphovascular space invasion (LVSI), assessed on hematoxylin and eosin-stained slides, is a major factor for adjuvant therapy in patients with cervical cancer. With the advent of a lymphatic endothelial cell-specific marker, such as D2-40, it is now possible to distinguish between blood and lymphatic space invasion (LSI). In this study, the utility of D2-40 was assessed for the detection of lymphatic vessel density (LVD) and identification of LSI. The expressions of vascular endothelial growth factor receptor-3 (VEGFR-3), VEGF-C, tyrosine receptor kinase-2, and angiopoietin-1 were assessed by immunohistochemical methods on 50 patients with squamous cell carcinoma of the cervix. Clinicopathologic characteristics, including pelvic lymph node metastasis, were correlated with the above histochemical findings. We found that lymphangiogenesis, measured by an increase in peritumoral LVD, was significantly associated with positive lymph node status (P < .005). VEGFR-3 expression was significantly associated with LVD (P < .05). D2-40 staining verified LSI (P = .03) and surpassed that of hematoxylin and eosin-identified LVSI (P = .54). In conclusion, lymphangiogenic markers, specifically LVD quantified by D2-40 and VEGFR-3, are independently associated with LSI and lymph node metastasis in patients with early squamous cell carcinoma of the cervix treated with radical hysterectomy and pelvic lymphadenectomy.


Mangiferin induces radiosensitization in glioblastoma cells by inhibiting nonhomologous end joining.
Although surgery and high‑dose radiotherapy have been the standard treatments for glioblastoma multiforme (GBM), these therapies are palliative, due to the high risk of local relapse. Emerging evidence has demonstrated that DNA double‑strand break (DSB) repair serves a critical role in resistance to radiotherapy. Previous studies have revealed that mangiferin possesses anti‑neoplastic effects on human lung adenocarcinoma and ovarian cancer. The present study aimed to investigate the role of mangiferin in radio‑sensitivity inhuman GBM. Through in vitro experiments, decreased proliferation and increased DNA damage were observed in cells pretreated with mangiferin following radiation. Further study of the repair pathway indicated that mangiferin inhibits the non‑homologous end‑joining (NHEJ) DSB repair pathway. Furthermore, studies on key proteins in the NHEJ DSB repair pathway revealed that mangiferin inhibited the phosphorylation of serine‑protein kinase ATM, TP53‑binding protein 1 and γ‑histone H2AX (γ‑H2AX). In addition, observations on the average percentages of γ‑H2AX‑positive cells and the average number of γ‑H2AX foci per cell suggested that treatment with mangiferin decreased the number of γ‑H2AX foci in GBM cells following radiation. However, mangiferin selectively inhibited DSB repair in GBM cells, and was not able to trigger DSB repair inhibition in normal neuronal Schwann cells. Through in vivo tumor‑bearing mouse experiments, a smaller tumor volume, decreased tumor weight and prolonged life span were observed in mice treated with mangiferin following radiation. Therefore, xenograft GBM models clearly demonstrated that treatment with mangiferin treatment may increase tumor sensitivity to radiotherapy. Taken together, as demonstrated by in vivo and in vitro data, mangiferin may be a potential novel therapeutic drug for improving the radiation sensitivity of glioblastoma.


Home-Based Gait Speed Assessment: Normative Data and Racial/Ethnic Correlates Among Older Adults.
To determine home-based gait speed performance and its associations with sociodemographic and health-related factors among older adults. Cross-sectional analysis of a nationally representative US population sample. Homes of Health and Retirement Study (HRS) participants. Walk test data measured at home over 2.5 m were aggregated for 6983 individuals, aged ≥65 years (mean age 74.8 ± 6.9 years, 54.2% women), from the 2006 and 2008 HRS waves. Means for gait speed at normal pace were determined for demographic and clinical groupings; association of gait speed with demographic, socioeconomic status, and health factors were examined. Four-year mortality was predicted from baseline slow gait status defined using demographic-based cutoff scores as well as commonly recommended cutoff scores (100 or 60 cm/s). Home-based gait speed (cm/s) means were lower for female than male (9.6% difference), older than younger (18.0% difference), African American than white (20.5% difference), and Hispanic than Non-Hispanic (10.3% difference) participants. Differences by age group, race, and ethnicity remained significant within sexes (P < .001). Lower speed was associated with African American race and all health problems; higher speed was associated with higher socioeconomic status and alcohol consumption. Four-year mortality was predicted by slow gait status. Predictive validity was, in general, higher for slow gait cutoff scores defined by demographic characteristics. Mean gait speed measured at home differs among older (aged ≥65 years) US resident population groups defined by sex, age, race, ethnicity, health status, and combinations of these factors, and predicts 4-year mortality when substantially slower than group-based norms. These findings may assist researchers and clinicians in determining normal and abnormal gait performance in older adults in community settings.


Pectoralis Plane Block for Pacemaker Insertion: A Successful Primary Anesthetic.
Effective anesthesia, analgesia, and hemodynamic stability is important to maintain during pacemaker implantation surgery, especially in the elderly population and patients with compromised cardiac function. As a strategy to avoid the need for intravenous (IV) anesthetics, peripheral nerve block techniques may be used in these specific cases. We report a case of successful pacemaker implantation surgery in a patient with severe Aortic Stenosis (AS) and Sick Sinus Syndrome (SSS) using unilateral pectoralis plane block for surgical anesthesia. Since general anesthesia was considered risky, monitored anesthesia care utilizing peripheral nerve block was planned. A single shot left side pectoralis plane block (PECS II) was done under ultrasound guidance injecting a total of 20 mL of 0.5% Ropivacaine with 1% Lidocaine. No sedation was needed. The patient tolerated the procedure with no significant hemodynamic changes. Patient did not require opioids post-operative and was discharged home in stable condition the next day. This case highlights that PECS block can also provide effective surgical anesthesia for relatively long procedures avoiding the risk of complications associated with IV anesthesia in high risk cardiovascular patients. Additionally, these blocks can provide an opioid sparing option for post-operative management in pacemaker implantation surgeries.


Biomechanical comparison of a modified three-loop pulley technique and a three-level self-locking technique for repair of patellar tendon rupture in dogs.
To compare use of a 3-level self-locking suture (3LSLS) technique with use of a previously described modified 3-loop pulley (M3LP) technique to repair rupture of the proximal aspect of patellar tendons in limbs from canine cadavers. Paired hind limbs of 6 adult mixed-breed dogs. A limb from each pair was randomly assigned to be repaired by the 3LSLS technique or M3LP technique with size-2 ultrahigh-molecular-weight polyethylene and size-0 monofilament polypropylene suture, respectively. Limbs were prepared, and each patellar tendon was transected at a site chosen to simulate rupture at the proximal aspect. Tendons were repaired with the assigned techniques and mechanically tested with a biaxial servohydraulic test system; the clamp was distracted until the repair failed. Force at 1 -mm gap formation, 3-mm gap formation, and repair failure and gap size at failure were measured and compared between methods. Mode of failure was recorded. There was no significant difference between methods for the force required to produce a 1 - or 3-mm gap in the repair. The 3LSLS technique required a significantly higher load for complete failure; gap formation immediately before failure was significantly greater for this method than for the M3LP technique. Four of 6 repairs with the M3LP technique and 0 of 6 repairs with the 3LSLS technique failed by suture breakage. The 3LSLS technique with size-2 ultrahigh-molecular-weight polyethylene suture was as effective at resistance to 1 - and 3-mm gap formation as the M3LP with size-0 monofilament polypropylene suture.


What to expect when you're evaluating healthcare improvement: a concordat approach to managing collaboration and uncomfortable realities.
Evaluation of improvement initiatives in healthcare is essential to establishing whether interventions are effective and to understanding how and why they work in order to enable replication. Although valuable, evaluation is often complicated by tensions and friction between evaluators, implementers and other stakeholders. Drawing on the literature, we suggest that these tensions can arise from a lack of shared understanding of the goals of the evaluation; confusion about roles, relationships and responsibilities; data burdens; issues of data flows and confidentiality; the discomforts of being studied and the impact of disappointing or otherwise unwelcome results. We present a possible approach to managing these tensions involving the co-production and use of a concordat. We describe how we developed a concordat in the context of an evaluation of a complex patient safety improvement programme known as Safer Clinical Systems Phase 2. The concordat development process involved partners (evaluators, designers, funders and others) working together at the outset of the project to agree a set of principles to guide the conduct of the evaluation. We suggest that while the concordat is a useful resource for resolving conflicts that arise during evaluation, the process of producing it is perhaps even more important, helping to make explicit unspoken assumptions, clarify roles and responsibilities, build trust and establish open dialogue and shared understanding. The concordat we developed established some core principles that may be of value for others involved in evaluation to consider. But rather than seeing our document as a ready-made solution, there is a need for recognition of the value of the process of co-producing a locally agreed concordat in enabling partners in the evaluation to work together effectively.


Disorders of the hallux sesamoid complex: MR features.
Numerous painful conditions can affect the first metatarsophalangeal-sesamoid joint complex. Symptoms can be of sudden or insidious onset, and be of acute or chronic duration. Although conventional radiography is recognized as the initial diagnostic procedure for these symptoms, there is often a need to proceed to MR imaging. MR imaging is sensitive and can be utilized in the investigation of the hallux sesamoid complex to differentiate soft tissue from osseous pathology. Synovitis, tendonitis, and bursitis can be distinguished from bony abnormalities such as sesamoid fracture, avascular necrosis, and osteomyelitis. An understanding of MR imaging features and techniques will result in the highest diagnostic yield. Early and accurate diagnosis of sesamoid complex disorders can guide the physician to the appropriate clinical management and prevent potentially harmful longstanding joint dysfunction.


Application of halloysite clay nanotubes as a pharmaceutical excipient.
Halloysite nanotubes, a biocompatible nanomaterial of 50-60nm diameter and ca. 15nm lumen, can be used for loading, storage and sustained release of drugs either in its pristine form or with additional polymer complexation for extended release time. This study reports the development composite tablets based on 50wt.% of the drug loaded halloysite mixed with 45wt.% of microcrystalline cellulose. Powder flow and compressibility properties of halloysite (angle of repose, Carr's index, Hausner ratio, Brittle Fracture Index, tensile strength) indicate that halloysite is an excellent tablet excipient. Halloysite tubes can also be filled with nifedipine with ca. 6wt.% loading efficiency and sustained release from the nanotubes. Tablets prepared with drug loaded halloysite allowed for almost zero order nifedipine release for up to 20h. Nifedipine trapped in the nanotubes also protect the drug against light and significantly increased the photostability of the drug. All of these demonstrate that halloysite has the potential to be an excellent pharmaceutical excipient that is also an inexpensive, natural and abundantly available material.


Ab initio calculation of inelastic scattering.
Nonresonant inelastic electron and X-ray scattering cross sections for bound-to-bound transitions in atoms and molecules are calculated directly from ab initio electronic wavefunctions. The approach exploits analytical integrals of Gaussian-type functions over the scattering operator, which leads to accurate and efficient calculations. The results are validated by comparison to analytical cross sections in H and He+, and by comparison to experimental results and previous theory for closed-shell He and Ne atoms, open-shell C and Na atoms, and the N2 molecule, with both inner-shell and valence electronic transitions considered. The method is appropriate for use in conjunction with quantum molecular dynamics simulations and for the analysis of new ultrafast X-ray scattering experiments.


A causal framework for understanding the effect of losses to follow-up on epidemiologic analyses in clinic-based cohorts: the case of HIV-infected patients on antiretroviral therapy in Africa.
Although clinic-based cohorts are most representative of the "real world," they are susceptible to loss to follow-up. Strategies for managing the impact of loss to follow-up are therefore needed to maximize the value of studies conducted in these cohorts. The authors evaluated adult patients starting antiretroviral therapy at an HIV/AIDS clinic in Uganda, where 29% of patients were lost to follow-up after 2 years (January 1, 2004-September 30, 2007). Unweighted, inverse probability of censoring weighted (IPCW), and sampling-based approaches (using supplemental data from a sample of lost patients subsequently tracked in the community) were used to identify the predictive value of sex on mortality. Directed acyclic graphs (DAGs) were used to explore the structural basis for bias in each approach. Among 3,628 patients, unweighted and IPCW analyses found men to have higher mortality than women, whereas the sampling-based approach did not. DAGs encoding knowledge about the data-generating process, including the fact that death is a cause of being classified as lost to follow-up in this setting, revealed "collider" bias in the unweighted and IPCW approaches. In a clinic-based cohort in Africa, unweighted and IPCW approaches-which rely on the "missing at random" assumption-yielded biased estimates. A sampling-based approach can in general strengthen epidemiologic analyses conducted in many clinic-based cohorts, including those examining other diseases.


Venous "supercharging" augments survival of the delayed rat TRAM flap.
Adequate delay of a pedicled transverse rectus abdominis musculocutaneous (TRAM) flap might not necessarily require interruption of the venous system. The retained ipsilateral deep vein of the dominant pedicle could then be used as a secondary outflow source for potential salvage of a congested flap. A venous "supercharged" rat TRAM flap model has been designed to evaluate the efficacy of this maneuver. Seventy-two female Sprague-Dawley rats (CD) were equally divided into two major groups, differing only in whether a delay by division of the dominant cranial epigastric artery had first been performed. An inferior-based TRAM flap (nondominant) was raised for each rat, with three subsets of 12 rats in each group, ie, with the cranial epigastric vein subsequently divided (group control), retained (supercharged), or retained but the inferior pedicle divided (venous flap). Both supercharged subsets had significantly augmented flap survival when compared with flaps in their group raised without the cranial epigastric vein, whether a delay maneuver had (96 +/- 6% vs 89 +/- 7%; P = 0.012) or not (80 +/- 8% vs 65 +/- 21%; P = 0.034) been performed. Flaps with only a cranial epigastric vein pedicle totally necrosed, implying that the observed enhancement in flap viability was not the result of transformation into a venous flap, but perhaps as a crossover flap where an adjacent venosome was captured. Venous supercharging can be accomplished by inclusion of the ipsilateral dominant deep vein, and should be a consideration in the clinical planning of delay maneuvers and for treatment of the compromised TRAM flap.


Efficacy of SERD/SERM Hybrid-CDK4/6 Inhibitor Combinations in Models of Endocrine Therapy-Resistant Breast Cancer.
Endocrine therapy, using tamoxifen or an aromatase inhibitor, remains first-line therapy for the management of estrogen receptor (ESR1)-positive breast cancer. However, ESR1 mutations or other ligand-independent ESR1 activation mechanisms limit the duration of response. The clinical efficacy of fulvestrant, a selective estrogen receptor downregulator (SERD) that competitively inhibits agonist binding to ESR1 and triggers receptor downregulation, has confirmed that ESR1 frequently remains engaged in endocrine therapy-resistant cancers. We evaluated the activity of a new class of selective estrogen receptor modulators (SERM)/SERD hybrids (SSH) that downregulate ESR1 in relevant models of endocrine-resistant breast cancer. Building on the observation that concurrent inhibition of ESR1 and the cyclin-dependent kinases 4 and 6 (CDK4/6) significantly increased progression-free survival in advanced patients, we explored the activity of different SERD- or SSH-CDK4/6 inhibitor combinations in models of endocrine therapy-resistant ESR1(+) breast cancer. SERDs, SSHs, and the CDK4/6 inhibitor palbociclib were evaluated as single agents or in combination in established cellular and animal models of endocrine therapy-resistant ESR1(+) breast cancer. The combination of palbociclib with a SERD or an SSH was shown to effectively inhibit the growth of MCF7 cell or ESR1-mutant patient-derived tumor xenografts. In tamoxifen-resistant MCF7 xenografts, the palbociclib/SERD or SSH combination resulted in an increased duration of response as compared with either drug alone. A SERD- or SSH-palbociclib combination has therapeutic potential in breast tumors resistant to endocrine therapies or those expressing ESR1 mutations. See related commentary by DeMichele and Chodosh, p. 4999.


Impact of vitamin E and C supplementation on serum adhesion molecules in chronic degenerative aortic stenosis: a randomized controlled trial.
An inflammatory component has been identified in degenerative aortic stenosis (AS). The combination of vitamins E and C has been shown to have anti-inflammatory properties. The aim of this study was to determine the impact of the combination of vitamins C and E or vitamin C only on serum levels of cell adhesion molecules and C-reactive protein in patients with chronic degenerative AS, with or without concomitant coronary artery disease. One hundred patients with asymptomatic or mildly symptomatic moderate AS were randomized in 2:2:1 format in an open-label trial. Forty-one patients received vitamin E (400 IU) and vitamin C (1000 mg) daily, 39 patients received vitamin C (1000 mg) only, and 20 patients were followed as controls. Serum intracellular adhesion molecule (ICAM-1), E selectin, P selectin, vascular-cellular adhesion molecule (VCAM-1), C-reactive protein, and alpha-tocopherol (vitamin E) were measured by enzyme-linked immunosorbent assay at baseline and 6 months postsupplementation. Half of the patients from each of the 2 active groups were followed for further 6 months to determine any changes after cessation of therapy. In the vitamin E and C, group there was reduction in serum ICAM-1 (298 +/- 12 to 272 +/- 12 ng/mL at 6 months, P = .0015) with a return to base line 6 months after cessation of therapy. In the vitamin C only group, there was a reduction in serum P selectin (134 +/- 10 to 118 +/- 10 ng/mL at 6 months, P = .033). All the inflammatory markers were unchanged in control group over 6 months of follow-up. Vitamin E and C supplementation had modest anti-inflammatory effect in chronic degenerative AS. The clinical relevance of this would require further clarification.


The Regenerative Role of the Fetal and Adult Stem Cell Secretome.
For a long time, the stem cell regenerative paradigm has been based on the assumption that progenitor cells play a critical role in tissue repair by means of their plasticity and differentiation potential. However, recent works suggest that the mechanism underlying the benefits of stem cell transplantation might relate to a paracrine modulatory effect rather than the replacement of affected cells at the site of injury. Therefore, mounting evidence that stem cells may act as a reservoir of trophic signals released to modulate the surrounding tissue has led to a paradigm shift in regenerative medicine. Attention has been shifted from analysis of the stem cell genome to understanding the stem cell "secretome", which is represented by the growth factors, cytokines and chemokines produced through paracrine secretion. Insights into paracrine-mediated repair support a new approach in regenerative medicine and the isolation and administration of specific stem cell-derived paracrine factors may represent an extremely promising strategy, introducing paracrine-based therapy as a novel and feasible clinical application. In this review, we will discuss the regenerative potential of fetal and adult stem cells, with particular attention to their secretome.


Chemical and magnetic analyses on tree bark as an effective tool for biomonitoring: A case study in Lisbon (Portugal).
Tree bark has proven to be a reliable tool for biomonitoring deposition of metals from the atmosphere. The aim of the present study was to test if bark magnetic properties can be used as a proxy of the overall metal loads of a tree bark, meaning that this approach can be used to discriminate different effects of pollution on different types of urban site. In this study, the concentrations of As, Cd, Co, Cu, Fe, Mn, Ni, P, Pb, V and Zn were measured by ICP-OES in bark samples of Jacaranda mimosifolia, collected along roads and in urban green spaces in the city of Lisbon (Portugal). Magnetic analyses were also performed on the same bark samples, measuring Isothermal Remanent Magnetization (IRM), Saturation Isothermal Remanent Magnetization (SIRM) and Magnetic Susceptibility (χ). The results confirmed that magnetic analyses can be used as a proxy of the overall load of trace elements in tree bark, and could be used to distinguish different types of urban sites regarding atmospheric pollution. Together with trace element analyses, magnetic analyses could thus be used as a tool to provide high-resolution data on urban air quality and to follow up the success of mitigation actions aiming at decreasing the pollutant load in urban environments.


Validity, Reliability, and Ability to Identify Fall Status of the Berg Balance Scale, BESTest, Mini-BESTest, and Brief-BESTest in Patients With COPD.
The Berg Balance Scale (BBS), Balance Evaluation Systems Test (BESTest), Mini-BESTest, and Brief-BESTest are useful in the assessment of balance. Their psychometric properties, however, have not been tested in patients with chronic obstructive pulmonary disease (COPD). This study aimed to compare the validity, reliability, and ability to identify fall status of the BBS, BESTest, Mini-BESTest, and the Brief-BESTest in patients with COPD. A cross-sectional study was conducted. Forty-six patients (24 men, 22 women; mean age=75.9 years, SD=7.1) were included. Participants were asked to report their falls during the previous 12 months and to fill in the Activity-specific Balance Confidence (ABC) Scale. The BBS and the BESTest were administered. Mini-BESTest and Brief-BESTest scores were computed based on the participants' BESTest performance. Validity was assessed by correlating balance tests with each other and with the ABC Scale. Interrater reliability (2 raters), intrarater reliability (48-72 hours), and minimal detectable changes (MDCs) were established. Receiver operating characteristics assessed the ability of each balance test to differentiate between participants with and without a history of falls. Balance test scores were significantly correlated with each other (Spearman correlation rho=.73-.90) and with the ABC Scale (rho=.53-.75). Balance tests presented high interrater reliability (intraclass correlation coefficient [ICC]=.85-.97) and intrarater reliability (ICC=.52-.88) and acceptable MDCs (MDC=3.3-6.3 points). Although all balance tests were able to identify fall status (area under the curve=0.74-0.84), the BBS (sensitivity=73%, specificity=77%) and the Brief-BESTest (sensitivity=81%, specificity=73%) had the higher ability to identify fall status. Findings are generalizable mainly to older patients with moderate COPD. The 4 balance tests are valid, reliable, and valuable in identifying fall status in patients with COPD. The Brief-BESTest presented slightly higher interrater reliability and ability to differentiate participants' fall status.


Application of the immunoregulatory receptor LILRB1 as a crystallisation chaperone for human class I MHC complexes.
X-ray crystallographic studies of class I peptide-MHC molecules (pMHC) continue to provide important insights into immune recognition, however their success depends on generation of diffraction-quality crystals, which remains a significant challenge. While protein engineering techniques such as surface-entropy reduction and lysine methylation have proven utility in facilitating and/or improving protein crystallisation, they risk affecting the conformation and biochemistry of the class I MHC antigen binding groove. An attractive alternative is the use of noncovalent crystallisation chaperones, however these have not been developed for pMHC. Here we describe a method for promoting class I pMHC crystallisation, by exploiting its natural ligand interaction with the immunoregulatory receptor LILRB1 as a novel crystallisation chaperone. First, focussing on a model HIV-1-derived HLA-A2-restricted peptide, we determined a 2.4 Å HLA-A2/LILRB1 structure, which validated that co-crystallisation with LILRB1 does not alter conformation of the antigenic peptide. We then demonstrated that addition of LILRB1 enhanced the crystallisation of multiple peptide-HLA-A2 complexes, and identified a generic condition for initial co-crystallisation. LILRB1 chaperone-based crystallisation enabled structure determination for HLA-A2 complexes previously intransigent to crystallisation, including both conventional and post-translationally-modified peptides, of diverse lengths. Since both the LILRB1 recognition interface on the HLA-A2 α3 domain molecule and HLA-A2-mediated crystal contacts are predominantly conserved across class I MHC molecules, the approach we outline could prove applicable to a diverse range of class I pMHC. LILRB1 chaperone-mediated crystallisation should expedite molecular insights into the immunobiology of diverse immune-related diseases and immunotherapeutic strategies, particularly involving class I pMHC complexes that are challenging to crystallise.


Managing Cancer Care During the COVID-19 Pandemic: Agility and Collaboration Toward a Common Goal.
The first confirmed case of coronavirus disease 2019 (COVID-19) in the United States was reported on January 20, 2020, in Snohomish County, Washington. At the epicenter of COVID-19 in the United States, the Seattle Cancer Care Alliance, Fred Hutchinson Cancer Research Center, and University of Washington are at the forefront of delivering care to patients with cancer during this public health crisis. This Special Feature highlights the unique circumstances and challenges of cancer treatment amidst this global pandemic, and the importance of organizational structure, preparation, agility, and a shared vision for continuing to provide cancer treatment to patients in the face of uncertainty and rapid change.


New insights into intestinal phages.
The intestinal microbiota plays important roles in human health. This last decade, the viral fraction of the intestinal microbiota, composed essentially of phages that infect bacteria, received increasing attention. Numerous novel phage families have been discovered in parallel with the development of viral metagenomics. However, since the discovery of intestinal phages by d'Hérelle in 1917, our understanding of the impact of phages on gut microbiota structure remains scarce. Changes in viral community composition have been observed in several diseases. However, whether these changes reflect a direct involvement of phages in diseases etiology or simply result from modifications in bacterial composition is currently unknown. Here we present an overview of the current knowledge in intestinal phages, their identity, lifestyles, and their possible effects on the gut microbiota. We also gather the main data on phage interactions with the immune system, with a particular emphasis on recent findings.


Preoptic mechanism for cold-defensive responses to skin cooling.
We recently identified a somatosensory pathway that transmits temperature information from the skin to a median subregion of the preoptic area (POA), a thermoregulatory centre. Here, we investigated in vivo the local neuronal circuit in the rat POA that processes the thermosensory information and outputs thermoregulatory effector signals. Skin cooling-evoked increases in sympathetic thermogenesis in brown adipose tissue, in metabolism and in heart rate were reversed by inhibition of neurons in the median preoptic nucleus (MnPO). Glutamatergic stimulation or disinhibition of MnPO neurons evoked thermogenic, metabolic and cardiac responses that mimicked the cold-defensive responses to skin cooling and were reversed by antagonizing GABA(A) receptors in the medial preoptic area (MPO), which is thought to contain neurons providing thermoregulatory output to effectors. These results suggest that GABA inhibition of output neurons in the MPO by MnPO neurons that are activated by cool sensory signals from the skin is a core thermoregulatory mechanism within the POA that is essential for the feedforward defence of body temperature against cold challenges in the environment.


SOST expression is restricted to the great arteries during embryonic and neonatal cardiovascular development.
Spatial-temporal regulation of bone morphogenetic protein (BMP) and Wnt activity is essential for normal cardiovascular development, and altered activity of these growth factors causes maldevelopment of the cardiac outflow tract and great arteries. In the present study, we show that SOST, a Dan family member reported to antagonize BMP and Wnt activity, is expressed within the medial vessel wall of the great arteries containing smooth muscle cells. The ascending aorta, aortic arch, brachiocephalic artery, common carotids, and pulmonary trunk were all associated with SOST expressing smooth muscle cells, while the heart itself, including the valves, and more distal arteries, that is, pulmonary arteries, subclavian arteries, and descending aorta, were negative. SOST was expressed from embryonic day 15.5 up to the neonatal period. SOST expression, however, did not correspond with inhibition of Smad-dependent BMP activity or beta-catenin-dependent Wnt activity in the great arteries. Activity of both signaling pathways was already down-regulated before induction of SOST expression.


Synaptophysin protein and mRNA expression in the human hippocampal formation from birth to old age.
In the human neocortex, progressive synaptogenesis in early postnatal life is followed by a decline in synaptic density, then stability from adolescence until middle age. No comparable data are available in the hippocampus. In this study, the integral synaptic vesicle protein synaptophysin, measured immunoautoradiographically, was used as an index of synaptic terminal abundance in the hippocampal formation of 37 subjects from 5 weeks to 86 yr old, divided into 4 age groups (10 infants, 15 adolescents/young adults, 6 adults, and 6 elderly). In all hippocampal subfields, synaptophysin was lowest in infancy, but did not differ significantly between the older age groups, except in dentate gyrus (DG) where the rise was delayed until adulthood. A similar developmental profile was found in the rat hippocampus. We also measured synaptophysin mRNA in the human subjects and found no age-related changes, except in parahippocampal gyrus wherein the mRNA declined from infancy to adolescence, and again in old age. The synaptophysin protein data demonstrate a significant presynaptic component to human postnatal hippocampal development. In so far as synaptophysin abundance reflects synaptic density, the findings support an increase in hippocampal and parahippocampal synapse formation during early childhood, but provide no evidence for adolescent synaptic pruning. The mRNA data indicate that the maturational increases in synaptophysin protein are either translational rather than transcriptional in origin, or else are secondary to mRNA increases in neurons, the cell bodies of which lie outside the hippocampal formation.