Estrogen mediation of breast tumor formation involves estrogen receptor-dependent, as well as independent, genotoxic effects.
Long-term exposure to estrogens influences the development of breast cancer in women, but the precise mechanisms involved are not clearly defined. Our working hypothesis is that estrogen modulates this process by two separate processes. One involves the binding of estradiol to estrogen receptor (ER) alpha with stimulation of cell proliferation. Errors in DNA occurring during replication result in fixed mutations when not repaired. The other process results from the formation of genotoxic metabolites of estradiol, which can bind to DNA, cause depurination, and result in mutations. Our collaborative group, funded by a Department of Defense Center of Excellence grant, has examined this hypothesis using a variety of experimental methods. Estradiol and its catechol-estrogen metabolite 4-OH-estradiol causes mutations in cell culture systems and can transform benign MCF-10F cells, allowing them to cause tumors in SCID mice. We have demonstrated loss of heterozygosity and gains and losses of DNA segments by comparative genomic hybridization methodology. The depurinated estradiol-guanine and -adenine adducts are measurable in MCF-7 breast cancer cells in culture and in mouse mammary tissue. The double transgenic, alpha estrogen receptor knockout/Wnt-1 knockin mouse model allows us to dissect out the separate effects of ER-mediated and ER-independent actions of estradiol. Knock out of the ER alpha delays the onset of breast tumors in this model, demonstrating a role of receptor-mediated actions. Oophorectomy delays the onset of tumors and reduces overall incidence, providing evidence for an ER-independent effect. Taken together, these data suggest that both ER-dependent and genotoxic ER-independent effects of estradiol mediate breast cancer development.


barx1 represses joints and promotes cartilage in the craniofacial skeleton.
The evolution of joints, which afford skeletal mobility, was instrumental in vertebrate success. Here, we explore the molecular genetics and cell biology that govern jaw joint development. Genetic manipulation experiments in zebrafish demonstrate that functional loss, or gain, of the homeobox-containing gene barx1 produces gain, or loss, of joints, respectively. Ectopic joints in barx1 mutant animals are present in every pharyngeal segment, and are associated with disrupted attachment of bone, muscles and teeth. We find that ectopic joints develop at the expense of cartilage. Time-lapse experiments suggest that barx1 controls the skeletal precursor cell choice between differentiating into cartilage versus joint cells. We discovered that barx1 functions in this choice, in part, by regulating the transcription factor hand2. We further show that hand2 feeds back to negatively regulate barx1 expression. We consider the possibility that changes in barx1 function in early vertebrates were among the key innovations fostering the evolution of skeletal joints.


Structural correlation method for model reduction and practical estimation of patient specific parameters illustrated on heart rate regulation.
We consider the inverse and patient specific problem of short term (seconds to minutes) heart rate regulation specified by a system of nonlinear ODEs and corresponding data. We show how a recent method termed the structural correlation method (SCM) can be used for model reduction and for obtaining a set of practically identifiable parameters. The structural correlation method includes two steps: sensitivity and correlation analysis. When combined with an optimization step, it is possible to estimate model parameters, enabling the model to fit dynamics observed in data. This method is illustrated in detail on a model predicting baroreflex regulation of heart rate and applied to analysis of data from a rat and healthy humans. Numerous mathematical models have been proposed for prediction of baroreflex regulation of heart rate, yet most of these have been designed to provide qualitative predictions of the phenomena though some recent models have been developed to fit observed data. In this study we show that the model put forward by Bugenhagen et al. can be simplified without loss of its ability to predict measured data and to be interpreted physiologically. Moreover, we show that with minimal changes in nominal parameter values the simplified model can be adapted to predict observations from both rats and humans. The use of these methods make the model suitable for estimation of parameters from individuals, allowing it to be adopted for diagnostic procedures.


Genetic counseling for hereditary cancer.
This paper provides an overview of the current approach to genetic counseling for cancer, using hereditary nonpolyposis colorectal cancer (HNPCC) as a prototype. Heretofore, when evaluating the possibility of an HNPCC diagnosis, physicians had to rely exclusively on a detailed family history of cancer in the context of an extended pedigree. Patients in the direct genetic lineage who had one or more first-degree relatives with an HNPCC syndrome cancer were told that they had a 50% likelihood of inheriting the deleterious gene. However, with the discovery of the HNPCC genes (hMSH2, hMLH1, hPMS1, hPMS2), genetic counseling can now provide a more precise determination of a patient's lifetime cancer destiny. Since these DNA findings are new, guidelines for sharing this information with patients remain preliminary. One must be certain that the patient wants to receive DNA information and that he or she is aware of potential discrimination by insurance companies and employers, as well as the possibility of psychological sequelae.


Regulation of endothelium-derived nitric oxide production by the protein kinase Akt.
Endothelial nitric oxide synthase (eNOS) is the nitric oxide synthase isoform responsible for maintaining systemic blood pressure, vascular remodelling and angiogenesis. eNOS is phosphorylated in response to various forms of cellular stimulation, but the role of phosphorylation in the regulation of nitric oxide (NO) production and the kinase(s) responsible are not known. Here we show that the serine/threonine protein kinase Akt (protein kinase B) can directly phosphorylate eNOS on serine 1179 and activate the enzyme, leading to NO production, whereas mutant eNOS (S1179A) is resistant to phosphorylation and activation by Akt. Moreover, using adenovirus-mediated gene transfer, activated Akt increases basal NO release from endothelial cells, and activation-deficient Akt attenuates NO production stimulated by vascular endothelial growth factor. Thus, eNOS is a newly described Akt substrate linking signal transduction by Akt to the release of the gaseous second messenger NO.


Sonoelectrochemical and sonochemical effects of cavitation: correlation with interfacial cavitation induced by 20 kHz ultrasound.
Sonoelectrochemical measurements at macro-electrodes under extreme conditions with a very short distance between ultrasonic horn tip and electrode and different ultrasound intensity levels are shown to result in violent cavitation detected in form of current peaks superimposed on the average limiting current. Analysis of the current data obtained for the oxidation of ferrocene in dimethylformamide (0.1 M NBu4PF6) at a 4 mm diameter Pt disc electrode and for the reduction of Ru(NH3)6(3+) in aqueous 0.1 M KCl at a 6 mm diameter Pt disc electrode consistently indicate a change of the physicochemical nature of sonoelectrochemical processes under extreme conditions. The sonoelectrochemical measurement of the rate constant for the carbon bromide bond cleavage of a 3-bromobenzophenone radical anion electrogenerated at a glassy carbon electrode in dimethylformamide solution in the presence of power ultrasound is shown to yield evidence for a breakdown of the conventional mass transport model of a planar diffusion layer under extreme conditions. The change can be correlated to the number of current data points deviating more than 10% from the mean of the current due to violent cavitation processes superimposed onto the average limiting current. Further, a study of the sonochemical destruction of aqueous dilute cyanide solution (in 0.1 M NaOH) demonstrates a correlation between the electrochemically detected cavitation violence and the sonochemical activity. Factors that govern the violence of interfacial cavitation appear to be directly proportional to the factors that make cavitation in the bulk solution chemically efficient.


Conceptualising the Integration of Strategies by Clinical Commissioning Groups in England towards the Antibiotic Prescribing Targets for the Quality Premium Financial Incentive Scheme: A Short Report.
Background: In order to tackle the public health threat of antimicrobial resistance, improvement in antibiotic prescribing in primary care was included as one of the priorities of the Quality Premium (QP) financial incentive scheme for Clinical Commissioning Groups (CCGs) in England. This paper briefly reports the outcome of a workshop exploring the experiences of antimicrobial stewardship (AMS) leads within CCGs in selecting and adopting strategies to help achieve the QP antibiotic targets. Methods: We conducted a thematic analysis of the notes on discussions and observations from the workshop to identify key themes. Results: Practice visits, needs assessment, peer feedback and audits were identified as strategies integrated in increasing engagement with practices towards the QP antibiotic targets. The conceptual model developed by AMS leads demonstrated possible pathways for the impact of the QP on antibiotic prescribing. Participants raised a concern that the constant targeting of high prescribing practices for AMS interventions might lead to disengagement by these practices. Most of the participants suggested that the effect of the QP might be less about the financial incentive and more about having national targets and guidelines that promote antibiotic prudency. Conclusions: Our results suggest that national targets, rather than financial incentives are key for engaging stakeholders in quality improvement in antibiotic prescribing.


Quantification of hemodynamics in abdominal aortic aneurysms during rest and exercise using magnetic resonance imaging and computational fluid dynamics.
Abdominal aortic aneurysms (AAAs) affect 5-7% of older Americans. We hypothesize that exercise may slow AAA growth by decreasing inflammatory burden, peripheral resistance, and adverse hemodynamic conditions such as low, oscillatory shear stress. In this study, we use magnetic resonance imaging and computational fluid dynamics to describe hemodynamics in eight AAAs during rest and exercise using patient-specific geometric models, flow waveforms, and pressures as well as appropriately resolved finite-element meshes. We report mean wall shear stress (MWSS) and oscillatory shear index (OSI) at four aortic locations (supraceliac, infrarenal, mid-aneurysm, and suprabifurcation) and turbulent kinetic energy over the entire computational domain on meshes containing more than an order of magnitude more elements than previously reported results (mean: 9.0-million elements; SD: 2.3 M; range: 5.7-12.0 M). MWSS was lowest in the aneurysm during rest 2.5 dyn/cm(2) (SD: 2.1; range: 0.9-6.5), and MWSS increased and OSI decreased at all four locations during exercise. Mild turbulence existed at rest, while moderate aneurysmal turbulence was present during exercise. During both rest and exercise, aortic turbulence was virtually zero superior to the AAA for seven out of eight patients. We postulate that the increased MWSS, decreased OSI, and moderate turbulence present during exercise may attenuate AAA growth.


Population growth and earth's human carrying capacity.
Earth's capacity to support people is determined both by natural constraints and by human choices concerning economics, environment, culture (including values and politics), and demography. Human carrying capacity is therefore dynamic and uncertain. Human choice is not captured by ecological notions of carrying capacity that are appropriate for nonhuman populations. Simple mathematical models of the relation between human population growth and human carrying capacity can account for faster-than-exponential population growth followed by a slowing population growth rate, as observed in recent human history.


Risk factors for device-associated infection related to organisational characteristics of intensive care units: findings from the Korean Nosocomial Infections Surveillance System.
Device-associated infections (DAIs) have been the major causes of morbidity and mortality of patients in intensive care units (ICUs). This study evaluated the risk factors for DAIs in ICUs. Ninety-six medical or surgical ICUs of 56 hospitals participated in the Korean Nosocomial Infections Surveillance System between July 2007 and June 2008. The occurrence of catheter-associated urinary tract infection (CAUTI), central line-associated bloodstream infection (CABSI), and ventilator-associated pneumonia (VAP) were monitored and DAI rates were calculated. Data associated with ICU characteristics were collected and Poisson regression was used for statistical analysis. Rates of CAUTI, CABSI, and VAP were 3.87 per 1000 urinary catheter days, 2.23 per 1000 central line days, and 1.89 per 1000 mechanical ventilator days, respectively. Rates of CAUTI were higher in ICUs in Seoul (P=0.032) and ICUs of major teaching hospitals (P=0.010). The ICUs of university-affiliated hospitals showed lower CAUTI rates (P=0.013). CABSI rates were higher in Seoul (P=0.001) and in medical ICUs (P=0.026). VAP rates were lower in ICUs of hospitals with more than 900 beds compared with hospitals with 400-699 beds (P=0.026). VAP rates were higher in surgical ICUs (P<0.0001) and increased 1.13-fold with each 100-unit increase in beds per infection control professional (P=0.003). The organisational and institutional characteristics of ICUs may influence DAI rates and there is a need for improvement in the incidence of VAP, CAUTI or CABSI.


High-reactivity matrices increase the sensitivity of matrix enhanced secondary ion mass spectrometry.
Secondary ion mass spectrometry (SIMS) is a desorption/ionization method in which ions are generated by the impact of a primary ion beam on a sample. Classic matrix assisted laser desorption and ionization (MALDI) matrices can be used to increase secondary ion yields and decrease fragmentation in a SIMS experiment, which is referred to as matrix enhanced SIMS (ME-SIMS). Contrary to MALDI, the choice of matrices for ME-SIMS is not constrained by their photon absorption characteristics. This implies that matrix compounds that exhibit an insufficient photon absorption coefficient have the potential of working well with ME-SIMS. Here, we evaluate a set of novel derivatives of the classical MALDI matrices α-cyano-4-hydroxycinnamic acid (CHCA) and 2,5-dihydroxybenzoic acid (DHB) for usability in ME-SIMS. This evaluation was carried out using peptide mixtures of different complexity and demonstrates significant improvements in signal intensity for several compounds with insufficient UV absorption at the standard MALDI laser wavelengths. Our study confirms that the gas-phase proton affinity of a matrix compound is a key physicochemical characteristic that determines its performance in a ME-SIMS experiment. As a result, these novel matrices improve the performance of matrix enhanced secondary ion mass spectrometry experiments on complex peptide mixtures.


A window of opportunity for reform in post-conflict settings? The case of Human Resources for Health policies in Sierra Leone, 2002-2012.
It is recognized that decisions taken in the early recovery period may affect the development of health systems. Additionally, some suggest that the immediate post-conflict period may allow for the opening of a political 'window of opportunity' for reform. For these reasons, it is useful to reflect on the policy space that exists in this period, by what it is shaped, how decisions are made, and what are their long-term implications. Examining the policy trajectory and its determinants can be helpful to explore the specific features of the post-conflict policy-making environment. With this aim, the study looks at the development of policies on human resources for health (HRH) in Sierra Leone over the decade after the conflict (2002-2012). Multiple sources were used to collect qualitative data on the period between 2002 and 2012: a stakeholder mapping workshop, a document review and a series of key informant interviews. The analysis draws from political economy and policy analysis tools, focusing on the drivers of reform, the processes, the contextual features, and the actors and agendas. Our findings identify three stages of policy-making. At first characterized by political uncertainty, incremental policies and stop-gap measures, the context substantially changed in 2009. The launch of the Free Health Care Initiative provided to be an instrumental event and catalyst for health system, and HRH, reform. However, after the launch of the initiative, the pace of HRH decision-making again slowed down. OUR STUDY IDENTIFIES THE KEY DRIVERS OF HRH POLICY TRAJECTORY IN SIERRA LEONE: (i) the political situation, at first uncertain and later on more defined; (ii) the availability of funding and the stances of agencies providing such funds; (iii) the sense of need for radical change - which is perhaps the only element related to the post-conflict setting. It also emerges that a 'windows of opportunity' for reform did not open in the immediate post-conflict, but rather 8 years later when the Free Health Care Initiative was announced, thus making it difficult to link it directly to the features of the post-conflict policy-making environment.


[Effects of chronic caloric restriction on the age-associated peculiarities of explorative behaviour in rats].
Long-term caloric restriction (CR) has been reported to extend the life spans, delay the onset and decrease the incidence of a broad spectrum of age-associated diseases. However, its effect on rat explorative behaviour is still unclear. In the present study, a number of behavioural measures were continuously monitored in 3-, 12-, 24-25-, 28-29- and 35-44-month-old male Wistar rats that were fed either ad libitum or placed on a caloric restricted diet. A gradual decline in locomotor activity of the ad libitum fed rats has been determined during aging in the open field test. In the CR groups, 3-month-old rats exhibited lower levels of exploratory behavior, compared to rats on the control diet. 24-25-month-old CR rats exhibited higher levels of exploratory behaviour, compared to ad libitum fed animals of the same age. Chronic dietary restriction nullified the age-dependent decline in locomotor activity and explorative behaviour of rats.


Partial Degradation of Recombinant Antibody Functional Activity During Infant Gastrointestinal Digestion: Implications for Oral Antibody Supplementation.
Oral administration of engineered immunoglobulins has the potential to prevent enteric pathogen-induced diarrhea in infants. To prevent infection, these antibodies need to survive functionally intact in the proteolytic environment of the gastrointestinal tract. This research examined both ex vivo and in vivo the functional survival across infant digestion of palivizumab, a model FDA-approved recombinant antibody against respiratory syncytial virus (RSV) F protein. Palivizumab-fortified feed (formula or human milk), infant gastric, and intestinal samples were incubated to simulate in vivo digestion (ex vivo digestion). Palivizumab-fortified human milk was also fed to infants, followed by collection of gastric and intestinal samples (in vivo digestion). Palivizumab was purified from the samples of digestate using protein G spin columns followed by filtration through molecular weight cut-off membranes (30 kDa). Palivizumab functional survival across ex vivo and in vivo digestion was determined via an anti-idiotype ELISA and an RSV plaque reduction neutralization test. Palivizumab concentration and RSV neutralization capacity both decreased when incubated in intestinal samples (ex vivo study). The concentration and neutralization activity of orally-supplemented palivizumab also decreased across infant digestion (in vivo study). These results indicate that if recombinant IgGs were selected for oral supplementation to prevent enteric infections, appropriate dosing would need to account for degradation occurring in the digestive system. Other antibody formats, structural changes, or encapsulation could enhance survival in the infant gastrointestinal tract.


Functional characterization of cell-to-cell coupling in cultured rat aortic smooth muscle.
Gap junction (GJ) occurrence and function was studied in cultured rat aortic smooth muscle cells, since cell-to-cell coupling is proposed to coordinate smooth muscle function but is difficult to study in the intact tissue. Cell proliferation in vitro formed a multilayered structure 10-15 cells thick. GJs connected cells to lateral and vertical neighbors, appearing in freeze fracture as P-face particles aggregated into circular plaques but also as linear arrays. The membrane potential was 58 +/- 3 mV. From quantification of the spread of electrotonic potentials according to a two-dimensional model, the intercellular resistivity was 900-1,400 omega X cm, whereas the nonjunctional membrane resistivity was 10(4) omega X cm2. Intercellular spread of 5(6)-carboxyfluorescein (CF; mol wt 376) in aortic cultures suggests that metabolic coupling is an important consequence of GJs in smooth muscle. CF transfer was not blocked by A23187 (10(-5) M), although rat fibroblasts became uncoupled by 10(-6) M. Ultimately uncoupled by the more potent ionophore ionomycin (10(-5) M), aortic cells seem more able to maintain GJ permeability during challenge from increased intracellular Ca than cells of noncontractile origin.


Expression of bcl-2 and Epstein-Barr virus LMP1 in lymphocytic interstitial pneumonia.
Epstein-Barr virus (EBV) genome has been demonstrated in lung tissues of patients with lymphocytic interstitial pneumonia (LIP) but its role in the pathogenesis of this condition is unclear. In vitro studies have shown that EBV can immortalise and transform cells by upregulation of the cellular proto-oncogene, B cell leukaemia-2 (bcl-2), via the viral latent membrane protein, LMP1. The purpose of this study was to determine whether bcl-2 expression is upregulated in the lungs of patients with LIP and whether EBV LMP1 has a role in this bcl-2 expression. Immunohistochemical analysis using alkaline phosphatase anti-alkaline phosphatase (APAAP) was performed on formalin fixed, paraffin embedded lung tissues from 13 patients with LIP using anti-LMP1 and anti-bcl-2 monoclonal antibodies. Lung tissues from nine patients with idiopathic pulmonary fibrosis (IPF) and nine necropsy cases without pulmonary disease served as controls. LMP1 positivity was estimated as the number of LMP1 positive cells per unit area of lung tissue. Immunostaining for bcl-2 expression was assessed by a pictorial-based semiquantitative grading system. Positive immunostaining for LMP1 was localised to airway epithelial cells of lung tissue. Ten out of 13 (77%) patients with LIP were positive for LMP1 compared with three of nine cases (33%) in each control group. LMP1 positivity of LIP cases was significantly greater than that of non-LIP cases: LIP versus IPF (mean difference, 95% confidence interval (CI)) 2.39 (1.54 to 3.24); LIP versus necropsy controls 2.62 (1.77 to 3.47). bcl-2 immunostaining was localised to lymphocytes within the alveolar septa and lymphoid aggregates of patients with LIP. The cumulative score for bcl-2 immunostaining was significantly higher in the lungs of patients with LIP than in those of patients with IPF and necropsy controls: LIP versus IPF and LIP versus necropsy controls (mean difference, 95% CI) 7.55 (7.18 to 7.92). These immunohistochemical studies have shown the presence of EBV LMP1 protein in airway epithelial cells and overexpression of the cellular bcl-2 protein in lymphoid cells of lung tissue in patients with LIP. These geographically distinct staining patterns of immunostaining suggest that the involvement of EBV LMP1 in the upregulation of cellular bcl-2 is more complex in LIP than was thought from previous in vitro observations. The respective roles of EBV LMP1 and bcl-2 in the pathogenesis of LIP require further studies.


Renal biopsy findings in Jamaican children.
To document the histological findings in Jamaican children undergoing renal biopsy in order to determine the relative prevalence of varying types of glomerular disease in the island. This study analyses retrospectively the renal histology in all Jamaican children less than age 12 years undergoing their first adequate renal biopsy between January 1985 and December 2008. Clinicopathological data were obtained solely from the histology reports from the University Hospital of the West Indies where all paediatric renal biopsies are processed. Of the 270 children, aged 1 month to 11 years (mean 7.58 years), 147 [58.1%] were males. The commonest indications for renal biopsy were nephrotic syndrome (57.4%) and glomerulonephritis (30%). Most biopsied children (260/270) had glomerular disease. The predominant glomerulonephritides were diffuse proliferative glomerulonephritis (DPGN) (27.7%) and mesangial proliferative glomerulonephritis (MesGN) (25.5%). Glomerular disease was idiopathic in 136/260 (53%) but was infection-associated in 32.3% (84 cases) of which Poststreptococcal glomerulonephritis (PSGN) was the commonest (75%)--predominantly DPGN (74.6%). Hepatitis B followed at 15.5% (13/84) manifested as membranous nephropathy in 83.3% (10/12). In patients with SS disease, DPGN was the commonest histology (47.4%). Systemic lupus erythematosus accounted for 5% of all renal biopsies. Over time, PSGN occurred less frequently, with a parallel reduction in DPGN and MesGN. In Jamaican children, DPGN is the commonest nephritis. Membranous nephropathy is primarily due to Hepatitis B. The commonest histology in SS disease is DPGN. The role of infection in the pathogenesis of renal disease in Jamaican children is probably underestimated.


Antimalarial natural products of marine and freshwater origin.
This review highlights recently discovered antimalarial natural products from marine and freshwater sources described in the literature from 2006 to 2008. The structures as well as bioactivities of compounds against the malaria parasites such as Plasmodium falciparum are discussed, including, for example, agelasine, xestoquinone, alisiaquinone, crambescidin, venturamide, dragomabin, gragonamide, viridamide, salinosporamide, chaetoxanthone, nodulisporacid, tumonoic acid, girolline, oroidin, nostocarboline, aerucyclamide, and microcylamide 7806 and its revised structure. Synthetic derivatives of natural products are presented including plakortin, isoaaptamine, curcuphenol, pseudopyronine, manzamine, and nostocarboline. Consequences of these discoveries for the development of novel natural product agents against malaria are discussed.


New Tools and Approaches for Family Physicians.
This issue of the journal is filled with useful information for practicing family physicians. Several articles introduce new ideas for family physicians to use in the care of their patients, whereas other articles cover new approaches to old problems. Several studies report on procedures performed by family physicians: battlefield acupuncture, colonoscopy, and ultrasound. Some unique alternative care models are described and evaluated. An innovative method of delivering diabetes education seems to work well. Ways to use technology to improve patient care, an update on chronic hepatitis B, and a novel use of social media to understand a rare disease are also included. Readers will come away from this issue with many ideas to consider implementing in their own practices.


Early-onset Tourette syndrome with reversible autistic behaviour: a dysmaturational disorder.
Early-onset Tourette syndrome comorbid with reversible autistic behaviour is described in twelve young males. After a normal gestation, delivery and first-year development, regression set in between the age of one and two with loss of various abilities and the emergence of autistic behaviour. At this time, or slightly later, they showed multiple motor and vocal tics, simple and complex: the latter could also be traced to most of their parents. Following an intervention based on intense cuddling, motor activation and paedagogic guidance, these children's abilities rapidly improved, reaching at follow-up a normal or borderline intellectual functioning and with the disappearance of their initial autistic behaviour. At follow-up tics were present in all, usually with the features of a full-blown Tourette syndrome, often comorbid with ADHD, and in some cases with OCD.


The coat morphogenetic protein SpoVID is necessary for spore encasement in Bacillus subtilis.
Endospores formed by Bacillus subtilis are encased in a tough protein shell known as the coat, which consists of at least 70 different proteins. We investigated the process of spore coat morphogenesis using a library of 40 coat proteins fused to green fluorescent protein and demonstrate that two successive steps can be distinguished in coat assembly. The first step, initial localization of proteins to the spore surface, is dependent on the coat morphogenetic proteins SpoIVA and SpoVM. The second step, spore encasement, requires a third protein, SpoVID. We show that in spoVID mutant cells, most coat proteins assembled into a cap at one side of the developing spore but failed to migrate around and encase it. We also found that SpoIVA directly interacts with SpoVID. A domain analysis revealed that the N-terminus of SpoVID is required for encasement and is a structural homologue of a virion protein, whereas the C-terminus is necessary for the interaction with SpoIVA. Thus, SpoVM, SpoIVA and SpoVID are recruited to the spore surface in a concerted manner and form a tripartite machine that drives coat formation and spore encasement.


Nutritional and metabolic status of children with autism vs. neurotypical children, and the association with autism severity.
The relationship between relative metabolic disturbances and developmental disorders is an emerging research focus. This study compares the nutritional and metabolic status of children with autism with that of neurotypical children and investigates the possible association of autism severity with biomarkers. Participants were children ages 5-16 years in Arizona with Autistic Spectrum Disorder (n = 55) compared with non-sibling, neurotypical controls (n = 44) of similar age, gender and geographical distribution. Neither group had taken any vitamin/mineral supplements in the two months prior to sample collection. Autism severity was assessed using the Pervasive Development Disorder Behavior Inventory (PDD-BI), Autism Treatment Evaluation Checklist (ATEC), and Severity of Autism Scale (SAS). Study measurements included: vitamins, biomarkers of vitamin status, minerals, plasma amino acids, plasma glutathione, and biomarkers of oxidative stress, methylation, sulfation and energy production. Biomarkers of children with autism compared to those of controls using a t-test or Wilcoxon test found the following statistically significant differences (p < 0.001): Low levels of biotin, plasma glutathione, RBC SAM, plasma uridine, plasma ATP, RBC NADH, RBC NADPH, plasma sulfate (free and total), and plasma tryptophan; also high levels of oxidative stress markers and plasma glutamate. Levels of biomarkers for the neurotypical controls were in good agreement with accessed published reference ranges. In the Autism group, mean levels of vitamins, minerals, and most amino acids commonly measured in clinical care were within published reference ranges.A stepwise, multiple linear regression analysis demonstrated significant associations between several groups of biomarkers with all three autism severity scales, including vitamins (adjusted R2 of 0.25-0.57), minerals (adj. R2 of 0.22-0.38), and plasma amino acids (adj. R2 of 0.22-0.39). The autism group had many statistically significant differences in their nutritional and metabolic status, including biomarkers indicative of vitamin insufficiency, increased oxidative stress, reduced capacity for energy transport, sulfation and detoxification. Several of the biomarker groups were significantly associated with variations in the severity of autism. These nutritional and metabolic differences are generally in agreement with other published results and are likely amenable to nutritional supplementation. Research investigating treatment and its relationship to the co-morbidities and etiology of autism is warranted.


Modafinil effects in multiple sclerosis patients with fatigue.
To investigate the effects of Modafinil on focused attention, motor function and motor excitability in patients with multiple sclerosis (MS) and fatigue. 21 MS patients with fatigue were enrolled in this double-blind placebo-controlled study. Modafinil (MOD) or placebo (PL) was administered for 8 weeks. The d2 alertness test, the Nine Hole Peg Test (9HPT) and several transcranial magnetic stimulation (TMS) techniques were applied prior to and after the first drug ingestion and well as after 8 weeks of drug intake. Prior to the first drug intake, the two groups were comparable. After the first drug ingestion, fatigue as measured by the Fatigue Severity Scale (FSS), performance of the d2 test and the 9HPT improved significantly in the MOD group and remained better than in the PL group after 8 weeks of treatment. Patients in the MOD group made fewer mistakes in the D2 test without being slower. They completed the 9HPT faster. Motor evoked potential amplitudes produced by paired pulse TMS were larger in the MOD group than the PL group. Motor thresholds and silent period durations remained unchanged. Compared to PL, MOD improved fatigue, focused attention and dexterity and enhanced motor cortex excitability in this group of patients. MOD may be helpful in MS patients with fatigue to improve cognitive and motor abilities.


Development and Psychometric Properties of the Taiwan Odd-Even Number Sequencing Test: A Nonalphabetic Measure of Working Memory.
Alphabetic working memory (WM) tests, such as the Wechsler Adult Intelligence Scale-III and IV Letter Number Sequencing, are not appropriate for nonalphabetic cultures. This study examined the psychometric properties of the Taiwan Odd-Even Number Sequencing Test (TOENST) and identified representative norms. The TOENST and other mental screening tasks were administered to 300 randomly selected healthy participants, 32 purposive sampling patients with schizophrenia, and 32 quota sampling controls. To investigate reliability and validity, a subset of the 300 healthy participants was randomly selected to receive a second TOENST ( n = 30) or conventional WM tests ( n = 42). The split-half reliability of the TOENST ranged from 0.69 to 0.95, and its test-retest reliability was 0.75. Criterion validity was demonstrated by significant correlations with conventional WM measures (all p < .05, except semantic verbal fluency), and construct validity was demonstrated by significant correlations with aging (main effect, F10,259 = 10.99, p < .001). Normative data were established, and performance was significantly associated with age and education. TOENST scores of patients with schizophrenia were significantly lower and correlated with frontal lobe tests, but not demographical or clinical characteristics. The TOENST has adequate psychometric properties and clinical utility and is as a viable alternative WM task for nonalphabetic cultures.


Criteria for the Research Institute for Fragrance Materials, Inc. (RIFM) safety evaluation process for fragrance ingredients.
The Research Institute for Fragrance Materials, Inc. (RIFM) has been engaged in the generation and evaluation of safety data for fragrance materials since its inception over 45 years ago. Over time, RIFM's approach to gathering data, estimating exposure and assessing safety has evolved as the tools for risk assessment evolved. This publication is designed to update the RIFM safety assessment process, which follows a series of decision trees, reflecting advances in approaches in risk assessment and new and classical toxicological methodologies employed by RIFM over the past ten years. These changes include incorporating 1) new scientific information including a framework for choosing structural analogs, 2) consideration of the Threshold of Toxicological Concern (TTC), 3) the Quantitative Risk Assessment (QRA) for dermal sensitization, 4) the respiratory route of exposure, 5) aggregate exposure assessment methodology, 6) the latest methodology and approaches to risk assessments, 7) the latest alternatives to animal testing methodology and 8) environmental risk assessment. The assessment begins with a thorough analysis of existing data followed by in silico analysis, identification of 'read across' analogs, generation of additional data through in vitro testing as well as consideration of the TTC approach. If necessary, risk management may be considered.


Adaptation of reach action to a novel force-field is not predicted by acuity of dynamic proprioception in either older or younger adults.
Healthy ageing involves degeneration of the neuromuscular system which impacts movement control and proprioception. Yet the relationship between these sensory and motor deficits in upper limb reaching has not been examined in detail. Recently, we reported that age-related proprioceptive deficits were unrelated to accuracy in rapid arm movements, but whether this applied in motor tasks more heavily dependent on proprioceptive feedback was not clear. To address this, we have tested groups of younger and older adults on a force-field adaptation task under either full or limited visual feedback conditions and examined how performance was related to dynamic proprioceptive acuity. Adaptive performance was similar between the age groups, regardless of visual feedback condition, although older adults showed increased after-effects. Physically inactive individuals made larger systematic (but not variable) proprioceptive errors, irrespective of age. However, dynamic proprioceptive acuity was unrelated to adaptation and there was no consistent evidence of proprioceptive recalibration with adaptation to the force-field for any group. Finally, in spite of clear age-dependent loss of spatial working memory capacity, we found no relationship between memory capacity and adaptive performance or proprioceptive acuity. Thus, non-clinical levels of deficit in dynamic proprioception, due to age or physical inactivity, do not affect force-field adaptation, even under conditions of limited visual feedback that might require greater proprioceptive control.


Adaptations in irrigated agriculture in the Mediterranean region: an overview and spatial analysis of implemented strategies.
In order to meet future food demand while sustainably managing available land and water resources, irrigated agriculture in semi-arid regions needs to adapt as a response to climate and socio-economic change. In this study, we focus on the Mediterranean region, a dynamic region, which is highly dependent on irrigated agriculture. We provide insight on adaptation strategies implemented on farm level, by doing a systematic review of studies in the region. Our analysis reports 286 implemented adaptations, on 124 different locations throughout the Mediterranean. Additionally, 142 drivers and 324 effects of adaptations were noted. We identified 31 adaptation strategies in 5 main categories: (1) water management, (2) sustainable resource management, (3) technological developments, (4) farm production practices, and (5) farm management. Strategies in the categories water management and farm production practices are most often implemented by farmers in the region. The main driver in the area is water scarcity and adaptations often affected water use and resources in addition to farm practices. Subsequently, we studied the spatial context of adaptations by analyzing the location factors of the five main strategies, using Geographic Information Systems and maximum entropy modeling. Our results show that farmers are more likely to adapt in less rural areas with lower poverty values and better market access, and in areas with higher temperatures and less rainfall. This demonstrates that both biophysical and socio-economic factors determine the context in which adaptations are implemented and that considerable spatial variability in the area exists.


The implementation of intentional rounding using participatory action research.
'Intentional'/'hourly rounding' is defined as regular checks of individual patients carried out by health professionals at set intervals rather than a response to a summons via a call bell. Intentional rounding places patients at the heart of the ward routine including the acknowledgement of patient preferences and in anticipation of their needs. The aim of this study was to implement intentional rounding using participatory action research to increase patient care, increase staff productivity and the satisfaction of care provision from both patients and staff. Outcomes of the study revealed a drop in call bell use, no observable threats to patient safety, nursing staff and patient satisfaction with care provision. However, any future studies should consider staff skill mix issues including the needs of newly graduated nursing staff as well as the cognitive status of patients when implementing intentional rounding on acute care wards.


Monoclonal antibodies in type 2 asthma: a systematic review and network meta-analysis.
Since novel treatments to target eosinophilic inflammation in Type 2 asthma are emerging, we aimed to evaluate and meta-analyze the efficacy of monoclonal antibodies to reduce exacerbation rate. PubMed and Web of Science were searched for phase II and phase III randomized clinical trials with monoclonal antibodies targeting key mediators of type 2-associated asthma. Thirty trials were selected involving biologics that target the IL-5 pathway, IL-13, the common IL-4 and IL-13 receptor, IL-9, IL-2 and TSLP. As no head-to-head trials were retrieved from literature, we performed an arm-based network meta-analysis to compare effects on exacerbation rate between the different treatments.Mepolizumab, reslizumab and benralizumab significantly reduced the risk of exacerbations compared to placebo (by 47-52%, 50-60%, and 28-51% respectively). Reslizumab and benralizumab also improved lung function. Dupilumab and tezepelumab improved lung function in frequent exacerbators. Lebrikizumab had no significant effect on the number of exacerbations, symptom control or health-related quality of life. Tralokinumab improved lung function compared to placebo. Network meta-analysis of all treatment and placebo arms, showed no superiority of one biologic over the others. Large reductions in exacerbation rates were observed compared to placebo, though only benralizumab was sufficiently powered (n = 2051) to demonstrate significantly decreased exacerbation rates in the subgroup analysis of IL-5 acting agents compared to placebo.Monoclonal antibodies such as mepolizumab, reslizumab and benralizumab have proven their benefit to reduce exacerbation rates in severe persistent eosinophilic asthma in the published trials. However, no statistically significant superiority was observed of one biologic over the other in the network meta-analysis. More studies with direct head to head comparisons and better defined endotypes are required.


Role of diabetologist in evaluating diabetic retinopathy.
To evaluate the ability of diabetologists to screen diabetic patients for diabetic retinopathy. Comparison of eye examination performed by diabetologists with direct ophthalmoscopy through an undilated pupil and by ophthalmologists through a dilated pupil with seven-field stereoscopic fundus photography (gold standard). The study consisted of 67 insulin-dependent and non-insulin-dependent diabetic outpatients attending a diabetes clinic. On the basis of fundus photography, patients were classified as having no or insignificant (30%), minimal (31%), moderate (24%), or severe (15%) retinopathy. The diabetologists and ophthalmologists performed similarly in their ability to classify severity of diabetic retinopathy accurately. When no or insignificant retinopathy (isolated microaneurysms only) was detected by examination, clinically significant retinopathy detected by fundus photography was highly unlikely (less than 5%). On the other hand, if more than isolated microaneurysms were seen on examination, all examiners missed more severe lesions detected by fundus photography. Patients with corrected visual acuity worse than 20/30 had a high likelihood (100%) of moderate or severe retinopathy. Motivated well-trained diabetologists can screen for diabetic retinopathy. The absence of detectable lesions by direct ophthalmoscopy indicates that automatic referral to an ophthalmologist is not necessary. However, if any level of retinopathy is detected or corrected acuity is worse than 20/30, referral to an ophthalmologist is required. In this setting, fundus photography is advised because it is the most sensitive means of detecting clinically significant retinopathy. If other nonophthalmologists can be trained to achieve similar results, current recommendations for ophthalmologic referral that require annual ophthalmologic examinations for most diabetic patients may need to be reconsidered.


Dynamic species classification of microorganisms across time, abiotic and biotic environments-A sliding window approach.
The development of video-based monitoring methods allows for rapid, dynamic and accurate monitoring of individuals or communities, compared to slower traditional methods, with far reaching ecological and evolutionary applications. Large amounts of data are generated using video-based methods, which can be effectively processed using machine learning (ML) algorithms into meaningful ecological information. ML uses user defined classes (e.g. species), derived from a subset (i.e. training data) of video-observed quantitative features (e.g. phenotypic variation), to infer classes in subsequent observations. However, phenotypic variation often changes due to environmental conditions, which may lead to poor classification, if environmentally induced variation in phenotypes is not accounted for. Here we describe a framework for classifying species under changing environmental conditions based on the random forest classification. A sliding window approach was developed that restricts temporal and environmentally conditions to improve the classification. We tested our approach by applying the classification framework to experimental data. The experiment used a set of six ciliate species to monitor changes in community structure and behavior over hundreds of generations, in dozens of species combinations and across a temperature gradient. Differences in biotic and abiotic conditions caused simplistic classification approaches to be unsuccessful. In contrast, the sliding window approach allowed classification to be highly successful, as phenotypic differences driven by environmental change, could be captured by the classifier. Importantly, classification using the random forest algorithm showed comparable success when validated against traditional, slower, manual identification. Our framework allows for reliable classification in dynamic environments, and may help to improve strategies for long-term monitoring of species in changing environments. Our classification pipeline can be applied in fields assessing species community dynamics, such as eco-toxicology, ecology and evolutionary ecology.


Photoactivated Spatiotemporally-Responsive Nanosensors of in Vivo Protease Activity.
Proteases play diverse and important roles in physiology and disease, including influencing critical processes in development, immune responses, and malignancies. Both the abundance and activity of these enzymes are tightly regulated and highly contextual; thus, in order to elucidate their specific impact on disease progression, better tools are needed to precisely monitor in situ protease activity. Current strategies for detecting protease activity are focused on functionalizing synthetic peptide substrates with reporters that emit detection signals following peptide cleavage. However, these activity-based probes lack the capacity to be turned on at sites of interest and, therefore, are subject to off-target activation. Here we report a strategy that uses light to precisely control both the location and time of activity-based sensing. We develop photocaged activity-based sensors by conjugating photolabile molecules directly onto peptide substrates, thereby blocking protease cleavage by steric hindrance. At sites of disease, exposure to ultraviolet light unveils the nanosensors to allow proteases to cleave and release a reporter fragment that can be detected remotely. We apply this spatiotemporally controlled system to probe secreted protease activity in vitro and tumor protease activity in vivo. In vitro, we demonstrate the ability to dynamically and spatially measure metalloproteinase activity in a 3D model of colorectal cancer. In vivo, veiled nanosensors are selectively activated at the primary tumor site in colorectal cancer xenografts to capture the tumor microenvironment-enriched protease activity. The ability to remotely control activity-based sensors may offer a valuable complement to existing tools for measuring biological activity.


The effect of food anticipation on cognitive function: An eye tracking study.
By randomizing the order in which participants perform a cognitive test and a food choice task in a controlled experiment, we investigate whether cognitive capacity can be enhanced by the simple act of anticipating food intake. Our findings show that overweight and obese participants exhibit an anticipatory food reward effect, which helped enhance their mental resources and improve their performance in a cognitive test. However, we find no anticipation effect among normal weight participants. Furthermore, eye tracking data reveal that food temptation, in the form of visual attention and emotional arousal is higher for overweight and obese individuals when they are cognitively impaired.


Brain activity during automatic semantic priming revealed by event-related functional magnetic resonance imaging.
Semantic priming occurs when a subject is faster in recognising a target word when it is preceded by a related word compared to an unrelated word. The effect is attributed to automatic or controlled processing mechanisms elicited by short or long interstimulus intervals (ISIs) between primes and targets. We employed event-related functional magnetic resonance imaging (fMRI) to investigate blood oxygen level dependent (BOLD) responses associated with automatic semantic priming using an experimental design identical to that used in standard behavioural priming tasks. Prime-target semantic strength was manipulated by using lexical ambiguity primes (e.g., bank) and target words related to dominant or subordinate meaning of the ambiguity. Subjects made speeded lexical decisions (word/nonword) on dominant related, subordinate related, and unrelated word pairs presented randomly with a short ISI. The major finding was a pattern of reduced activity in middle temporal and inferior prefrontal regions for dominant versus unrelated and subordinate versus unrelated comparisons, respectively. These findings are consistent with both a dual process model of semantic priming and recent repetition priming data that suggest that reductions in BOLD responses represent neural priming associated with automatic semantic activation and implicate the left middle temporal cortex and inferior prefrontal cortex in more automatic aspects of semantic processing.


Disparities in maternal outcomes among four ethnic populations.
To examine racial disparities in adverse maternal outcomes among four ethnic groups in the United States. A total of 1,030,350 women aged 13 to 55 who delivered in 1998 and 1999. Data were abstracted from the National Inpatient Sample. Race was categorized as White, African-American, Hispanic, and Asian/Pacific Islander. Maternal outcome measures included preterm labor, hypertensive disorders of pregnancy, gestational diabetes, antepartum hemorrhage, membrane disorders, cesarean section, and postpartum hemorrhage. African Americans were more likely to have preterm labor (odds ratio [OR] 1.71), preeclampsia (OR 1.59), transient hypertension of pregnancy (THP) (OR 1.13), pregnancy-induced hypertension (PIH) (OR 1.38), diabetes (OR 1.26), placenta previa (OR 1.78), placental abruption (OR 1.52), premature rupture of membranes (PRM) (OR 1.19), infection of the amniotic cavity (IAC) (OR 1.95), and cesarean section (risk ratio [RR] 1.08); Hispanics were more likely to have diabetes (OR 1.44), placenta previa (OR 1.20), IAC (OR 1.15), and cesarean section (RR 1.06); and Asian/Pacific Islanders were more likely to have diabetes (OR 2.05), placenta previa (OR 1.57), PRM (OR 1.26), IAC (1.79), and postpartum hemorrhage (OR 1.19). African Americans had the worst outcomes when adjusted for sociodemographic characteristics and comorbidities. Infection, gestational diabetes, and control of hypertension, especially among African Americans, are the most significant preventable risk factors contributing to disparities. By increasing access to health services, health education, and appropriate perinatal care in minority populations, improvements are likely to occur across multiple measures including frequencies of preterm labor, cesarean section, and gestational diabetes.


A plan to prevent adolescent pregnancy and reduce infant mortality.
In Illinois, particularly in Chicago, the infant mortality rate is higher than that for the nation as a whole. Unless infant mortality in Illinois declines at a faster rate, the State will not meet the objective for reducing infant mortality specified by the Surgeon General of the Public Health Service. Low birth weight infants are at high risk of medical problems, if not death, and teenage mothers have an increased risk of bearing low birth weight infants. This paper presents a program aimed at the primary prevention of adolescent pregnancy. The three-component, interactive program--sex education, adolescent clinic services, and community support and involvement--is proposed for preventing adolescent pregnancy which, in turn, will decrease the rate of births of low birth weight infants and, consequently, the rate of infant mortality in target areas of Illinois. Medical and college students will be trained to teach the sex education course and to serve as counselors. A clinic will provide primary care, contraceptive education, and access to contraceptives. The clinic staff will be a resource for the community as well as for the adolescents. In the target areas, church and community members will sponsor health fairs and discussions of adolescent pregnancy at church and at parent-teacher association meetings. The overlapping activities of the instructors, clinic personnel, and church and community members will give continuity and consistency to the prevention message directed to the adolescents.


Structure of the TatC core of the twin-arginine protein transport system.
The twin-arginine translocation (Tat) pathway is one of two general protein transport systems found in the prokaryotic cytoplasmic membrane and is conserved in the thylakoid membrane of plant chloroplasts. The defining, and highly unusual, property of the Tat pathway is that it transports folded proteins, a task that must be achieved without allowing appreciable ion leakage across the membrane. The integral membrane TatC protein is the central component of the Tat pathway. TatC captures substrate proteins by binding their signal peptides. TatC then recruits TatA family proteins to form the active translocation complex. Here we report the crystal structure of TatC from the hyperthermophilic bacterium Aquifex aeolicus. This structure provides a molecular description of the core of the Tat translocation system and a framework for understanding the unique Tat transport mechanism.


Rising oceans, climate change, food aid, and human rights in the Marshall Islands.
Climate change impacts are expected to produce more frequent, longer and unpredictable drought periods with further saltwater intrusion in the Marshall Islands. As a result, a significant return to traditional food cropping is unlikely. This will lead to an increased dependence on food aid, especially in the outer atoll populations. An examination of the nutritional content of food aid suggests it is likely to lead to poor health outcomes. Dependence on food aid has gradually increased over the past 70 years in the Marshall Islands, starting with population relocation because of war and nuclear testing and most recently because of climate change. The authors argue that the health impacts of the supplemental imported diet, combined with migration to population centers, may result in an even greater prevalence of chronic diseases, and exert pressures that lead to more communicable disease, further exacerbating the syndemics in the Marshall Islands. The authors conclude that food aid donors and the Republic of the Marshall Islands (RMI) government have human rights obligations to ensure that the people in the Marshall Islands have access to adequate nutrition. Accordingly, donors and the government should re-examine the content of food and ensure it is of sufficient quality to meet the right to health obligations.


Untargeted metabolomic analysis of human plasma indicates differentially affected polyamine and L-arginine metabolism in mild cognitive impairment subjects converting to Alzheimer's disease.
This study combined high resolution mass spectrometry (HRMS), advanced chemometrics and pathway enrichment analysis to analyse the blood metabolome of patients attending the memory clinic: cases of mild cognitive impairment (MCI; n = 16), cases of MCI who upon subsequent follow-up developed Alzheimer's disease (MCI_AD; n = 19), and healthy age-matched controls (Ctrl; n = 37). Plasma was extracted in acetonitrile and applied to an Acquity UPLC HILIC (1.7μm x 2.1 x 100 mm) column coupled to a Xevo G2 QTof mass spectrometer using a previously optimised method. Data comprising 6751 spectral features were used to build an OPLS-DA statistical model capable of accurately distinguishing Ctrl, MCI and MCI_AD. The model accurately distinguished (R2 = 99.1%; Q2 = 97%) those MCI patients who later went on to develop AD. S-plots were used to shortlist ions of interest which were responsible for explaining the maximum amount of variation between patient groups. Metabolite database searching and pathway enrichment analysis indicated disturbances in 22 biochemical pathways, and excitingly it discovered two interlinked areas of metabolism (polyamine metabolism and L-Arginine metabolism) were differentially disrupted in this well-defined clinical cohort. The optimised untargeted HRMS methods described herein not only demonstrate that it is possible to distinguish these pathologies in human blood but also that MCI patients 'at risk' from AD could be predicted up to 2 years earlier than conventional clinical diagnosis. Blood-based metabolite profiling of plasma from memory clinic patients is a novel and feasible approach in improving MCI and AD diagnosis and, refining clinical trials through better patient stratification.


Assembly of a chromosomal replication machine: two DNA polymerases, a clamp loader, and sliding clamps in one holoenzyme particle. I. Organization of the clamp loader.
The gamma complex of DNA polymerase III holoenzyme, the replicase of Escherichia coli, couples ATP hydrolysis to the loading of beta sliding clamps onto primed DNA. The beta sliding clamp tethers the holoenzyme replicase to DNA for rapid and processive synthesis. In this report, the gamma complex has been constituted from its five different subunits. Size measurements and subunit stoichiometry studies show a composition of gamma 2 delta 1 delta' 1 1 chi 1 psi 1. Strong intersubunit contacts have been identified by gel filtration, and weaker contacts were identified by surface plasmon resonance measurements. An analogous tau complex has also been constituted and characterized; it is nearly as active as the gamma complex in clamp loading activity, but as shown in the fourth report of this series, it is at a disadvantage in binding the delta, delta', chi, and psi subunits when core is present (Xiao, H., Naktinis, V., and O'Donnell, M. (1995) J. Biol. Chem. 270, 13378-13383). The single copy subunits within the gamma complex provide the basis for the structural asymmetry inherent within DNA polymerase III holoenzyme.


The broad spectrum of autoimmune lymphoproliferative disease: molecular bases, clinical features and long-term follow-up in 31 patients.
Autoimmune lymphoproliferative disorders, including autoimmune lymphoproliferative syndrome (ALPS) and Dianzani autoimmune lymphoproliferative disease (DALD), are inherited defects of the Fas apoptotic pathway characterized by lymphoid accumulation and autoimmune manifestations. We report the molecular, clinical, immunologic features and the long-term progress of 31 patients. Four carried Fas gene mutations and one also displayed a caspase 10 polymorphism that probably contributed to the phenotype. Seven patients developed antibody deficiency and their clinical pictures overlapped those of subjects with common variable immunodeficiency (CVID). We postulate the existence of a disorder that involves the Fas pathway and displays the characteristics of both autoimmune lymphoproliferative disease and CVID.


IRS2 increases mitochondrial dysfunction and oxidative stress in a mouse model of Huntington disease.
Aging is a major risk factor for the progression of neurodegenerative diseases, including Huntington disease (HD). Reduced neuronal IGF1 or Irs2 signaling have been shown to extend life span in mice. To determine whether Irs2 signaling modulates neurodegeneration in HD, we genetically modulated Irs2 concentrations in the R6/2 mouse model of HD. Increasing Irs2 levels in the brains of R6/2 mice significantly reduced life span and increased neuronal oxidative stress and mitochondrial dysfunction. In contrast, reducing Irs2 levels throughout the body (except in β cells, where Irs2 expression is needed to prevent diabetes onset; R6/2•Irs2+/-•Irs2βtg mice) improved motor performance and extended life span. The slower progression of HD-like symptoms was associated with increased nuclear localization of the transcription factor FoxO1 and increased expression of FoxO1-dependent genes that promote autophagy, mitochondrial function, and resistance to oxidative stress. Mitochondrial function improved and the number of autophagosomes increased in R6/2•Irs2+/-•Irs2βtg mice, whereas aggregate formation and oxidative stress decreased. Thus, our study suggests that Irs2 signaling can modulate HD progression. Since we found the expression of Irs2 to be normal in grade II HD patients, our results suggest that decreasing IRS2 signaling could be part of a therapeutic approach to slow the progression of HD.


Children's understanding of the implications of causal attributions for emotional experiences.
Children's perceptions of how the cause of achievement outcomes affects individuals' emotional responses were studied. In Study 1, children aged 6 and 7, 9 and 10, and 12 and 13 listened to stories describing hypothetical children's achievement outcomes. Success and failure were explicitly attributed to luck, ability, effort, or another person's intervention. After each story subjects rated the story child's emotional reactions. Only seventh graders associated pride and shame exclusively with outcomes attributed to ability and effort. Guilt was strongly associated with effort attributions, and surprise was associated with luck attributions for fourth- and seventh-grade children but not for first-grade children. The attribution-affect linkages assumed by the older children are the same as those found in previous studies of adults. In Study 2, children aged 6 and 7, 9 and 10, and 12 and 13 rated the cause of the outcomes in the same stories according to Weiner's controllability and locus dimensions. Children's placement of specific attributions on these dimensions was used to explain age differences in their beliefs about the effect of the attributions on emotional responses.


Orbits in the H(2)O molecule.
We study the forms of the orbits in a symmetric configuration of a realistic model of the H(2)O molecule with particular emphasis on the periodic orbits. We use an appropriate Poincare surface of section (PSS) and study the distribution of the orbits on this PSS for various energies. We find both ordered and chaotic orbits. The proportion of ordered orbits is almost 100% for small energies, but decreases abruptly beyond a critical energy. When the energy exceeds the escape energy there are still nonescaping orbits around stable periodic orbits. We study in detail the forms of the various periodic orbits, and their connections, by providing appropriate stability and bifurcation diagrams. (c) 2001 American Institute of Physics.


Fc γ R IIB gene polymorphisms in Indian systemic lupus erythematosus (SLE) patients.
Receptors for the Fc fragment of immunoglobulin G (Fc γ Rs) represent the link between humoral and cellular immune responses. Polymorphisms in Fc γ Rs have been identified as genetic factors influencing susceptibility to various autoimmune diseases. This study was aimed to identify Fc γ R IIB genotypes in Indian systemic lupus erythematosus (SLE) patients and to correlate these with clinical presentation and autoantibody profile. Eighty consecutive clinically diagnosed SLE patients were included. SLE patients were classified according to the American College of Rheumatology (ACR) criteria. Disease activity was assessed by Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). PCR-RFLP method was used to detect Fc γ R IIB polymorphism. Of the 80 SLE patients, 53 were LN and 27 were SLE without nephritis. The mean SLEDAI score at evaluation was 6.5 ± 5.8. Among SLE patients Fc γ R IIB genotype frequency was 61.2 per cent for Ile/Thr, 20.0 per cent for Thr/Thr and 18.8 per cent for Ile/Ile as compared to 65, 12.5 and 22.5 per cent respectively among normal population. There was no significant difference for Fc γ R IIB genotypes between SLE and normals. The allele frequency for Thr allele in SLE patients was slightly higher (0.51) than in normals (0.45). Thr allele frequency in LN patients was slightly higher (0.53) than in SLE patients without nephritis (0.49). Though a higher percentages of symptoms like renal manifestations (81.3%), arthritis (62.5%) and oral ulcer (56.3%) were noted in patients with Thr/Thr genotypes, no significant difference was noted when these patients were compared with Ile/Ile and Ile/Thr genotypes. The findings of this study indicate towards an involvement of Thr allele with SLE disease severity and clinical presentation in Indian SLE patients. Future study on a large sample is needed to support this finding to understand the association of Fc γ R IIB 232Thr/Thr genotype as a susceptibility factor in SLE.


Sex Differences in the Relationship Between Left Ventricular Diastolic Dysfunction and Coronary Artery Disease: From the Korean Women's Chest Pain Registry.
Although coronary artery disease (CAD) is an important comorbidity, the influence of CAD on left ventricular diastolic dysfunction (LVDD) is not clear. This study aims to assess differences in the relationship between CAD and LVDD according to sex and comorbidities. A total of 1109 consecutive patients who visited an outpatient clinic with chest pain were included. The outcomes in 295 men and 295 women were compared after propensity score 1:1 matching. The evaluation of LVDD was based on guidelines recommended by the American Society of Echocardiography. The presence and severity of obstructive CAD was assessed by coronary angiography. After propensity score matching, both men and women were well balanced, with no significant differences of baseline covariates. Differences in diastolic functional parameters according to CAD severity were observed only in women (three parameters of LVDD except e' velocity, p < 0.05). Multiple logistic regression for matched pair data demonstrated that CAD severity was associated with the presence of LVDD only in women (odds ratio 1.919, 95% confidence interval 1.343-2.741, p < 0.001). Interaction between sex and severity of CAD was significant (p = 0.025). The association of LVDD and CAD severity was observed only in women. Myocardial ischemia may be a potential pathophysiology for higher prevalence of LVDD and heart failure with preserved ejection fraction in women.


Kinetic and structural analysis of human ALDH9A1.
Aldehyde dehydrogenases (ALDHs) constitute a superfamily of NAD(P)+-dependent enzymes, which detoxify aldehydes produced in various metabolic pathways to the corresponding carboxylic acids. Among the 19 human ALDHs, the cytosolic ALDH9A1 has so far never been fully enzymatically characterized and its structure is still unknown. Here, we report complete molecular and kinetic properties of human ALDH9A1 as well as three crystal forms at 2.3, 2.9, and 2.5 Å resolution. We show that ALDH9A1 exhibits wide substrate specificity to aminoaldehydes, aliphatic and aromatic aldehydes with a clear preference for γ-trimethylaminobutyraldehyde (TMABAL). The structure of ALDH9A1 reveals that the enzyme assembles as a tetramer. Each ALDH monomer displays a typical ALDHs fold composed of an oligomerization domain, a coenzyme domain, a catalytic domain, and an inter-domain linker highly conserved in amino-acid sequence and folding. Nonetheless, structural comparison reveals a position and a fold of the inter-domain linker of ALDH9A1 never observed in any other ALDH so far. This unique difference is not compatible with the presence of a bound substrate and a large conformational rearrangement of the linker up to 30 Å has to occur to allow the access of the substrate channel. Moreover, the αβE region consisting of an α-helix and a β-strand of the coenzyme domain at the dimer interface are disordered, likely due to the loss of interactions with the inter-domain linker, which leads to incomplete β-nicotinamide adenine dinucleotide (NAD+) binding pocket.


Self-Paced Balance Learning for Clinical Skin Disease Recognition.
Class imbalance is a challenging problem in many classification tasks. It induces biased classification results for minority classes that contain less training samples than others. Most existing approaches aim to remedy the imbalanced number of instances among categories by resampling the majority and minority classes accordingly. However, the imbalanced level of difficulty of recognizing different categories is also crucial, especially for distinguishing samples with many classes. For example, in the task of clinical skin disease recognition, several rare diseases have a small number of training samples, but they are easy to diagnose because of their distinct visual properties. On the other hand, some common skin diseases, e.g., eczema, are hard to recognize due to the lack of special symptoms. To address this problem, we propose a self-paced balance learning (SPBL) algorithm in this paper. Specifically, we introduce a comprehensive metric termed the complexity of image category that is a combination of both sample number and recognition difficulty. First, the complexity is initialized using the model of the first pace, where the pace indicates one iteration in the self-paced learning paradigm. We then assign each class a penalty weight that is larger for more complex categories and smaller for easier ones, after which the curriculum is reconstructed by rearranging the training samples. Consequently, the model can iteratively learn discriminative representations via balancing the complexity in each pace. Experimental results on the SD-198 and SD-260 benchmark data sets demonstrate that the proposed SPBL algorithm performs favorably against the state-of-the-art methods. We also demonstrate the effectiveness of the SPBL algorithm's generalization capacity on various tasks, such as indoor scene image recognition and object classification.


Seasonal nitrogen fluxes of the Lena River Delta.
The Arctic is nutrient limited, particularly by nitrogen, and is impacted by anthropogenic global warming which occurs approximately twice as fast compared to the global average. Arctic warming intensifies thawing of permafrost-affected soils releasing their large organic nitrogen reservoir. This organic nitrogen reaches hydrological systems, is remineralized to reactive inorganic nitrogen, and is transported to the Arctic Ocean via large rivers. We estimate the load of nitrogen supplied from terrestrial sources into the Arctic Ocean by sampling in the Lena River and its Delta. We took water samples along one of the major deltaic channels in winter and summer in 2019 and sampling station in the central delta over a one-year cycle. Additionally, we investigate the potential release of reactive nitrogen, including nitrous oxide from soils in the Delta. We found that the Lena transported nitrogen as dissolved organic nitrogen to the coastal Arctic Ocean and that eroded soils are sources of reactive inorganic nitrogen such as ammonium and nitrate. The Lena and the Deltaic region apparently are considerable sources of nitrogen to nearshore coastal zone. The potential higher availability of inorganic nitrogen might be a source to enhance nitrous oxide emissions from terrestrial and aquatic sources to the atmosphere.


Photon statistics and correlation analysis of ultraweak light originating from living organisms for extraction of biological information.
Ultraweak photon emission phenomena in the visible to near-IR region, originating from biological organisms, are known. This biophoton emission is generated during metabolic processes and constitutes physiological information. We investigated a technique for characterizing the optical radiation field based on photon statistics and correlation analysis to extract information on regulation processes in biochemical reactions and their interactions. We developed the system based on the time-interval measurement of photoelectrons in a photon-counting region and employed data processing with a nonstationary optical field with correction for the correlative properties of the photomultiplier dark current. We analyzed biophoton emission from cellular slime mold (Dictyosterium discoideum) and observed the characteristic variation of this organism's super-Poisson statistics during the developmental process.


Early platelet consumption in pre-eclampsia.
One hundred and thirty-one women with chronic hypertension were studied serially during pregnancy to determine the sequence of events in the development of superimposed pre-eclampsia and to discover the time of onset. Twenty-seven women developed a sustained rise in plasma urate concentrations, which began at about 28 weeks' gestation and which is characteristic of pre-eclampsia. The mean platelet count was already significantly reduced and continued to fall until delivery, which was on average at 36 weeks' gestation. A comparable but smaller decrease in platelet count was seen in 55 women who had borderline but consistent increases in plasma urate concentrations. In 49 women whose plasma urate concentrations remained steady the platelet count did not change significantly before delivery. The reduced platelet count in women who develop pre-eclampsia suggests that increased platelet consumption is an early feature of the disorder.


Dose-related effects of delta-9-THC on emotional responses to acute psychosocial stress.
Cannabis smokers often report that they use the drug to relax or to relieve emotional stress. However, few clinical studies have shown evidence of the stress-relieving effects of cannabis or cannabinoid agonists. In this study, we sought to assess the influence of delta-9-tetrahydrocannabinol (THC), a main active ingredient of cannabis, upon emotional responses to an acute psychosocial stressor among healthy young adults. Healthy volunteers (N=42) participated in two experimental sessions, one with psychosocial stress (Trier Social Stress Test, TSST) and another with a non-stressful task, after receiving 0 (N=13), 7.5mg (N=14) or 12.5mg (N=15) oral THC. Capsules were administered under randomized, double blind conditions, 2.5h before the tasks began. We measured subjective mood and drug effects, vital signs and salivary cortisol before and at repeated times after the capsule and tasks. Subjects also appraised the tasks, before and after completion. In comparison to placebo, 7.5mg THC significantly reduced self-reported subjective distress after the TSST and attenuated post-task appraisals of the TSST as threatening and challenging. By contrast, 12.5mg THC increased negative mood overall i.e., both before and throughout the tasks, and pre-task ratings of the TSST as threatening and challenging. It also impaired TSST performance and attenuated blood pressure reactivity to the stressor. Our findings suggest that a low dose of THC produces subjective stress-relieving effects in line with those commonly reported among cannabis users, but that higher doses may non-specifically increase negative mood.


Fetal monitoring indications for delivery and 2-year outcome in 310 infants with fetal growth restriction delivered before 32 weeks' gestation in the TRUFFLE study.
In the TRUFFLE (Trial of Randomized Umbilical and Fetal Flow in Europe) study on the outcome of early fetal growth restriction, women were allocated to one of three groups of indication for delivery according to the following monitoring strategies: (1) reduced fetal heart rate (FHR) short-term variation (STV) on cardiotocography (CTG); (2) early changes in fetal ductus venosus (DV) waveform (DV-p95); and (3) late changes in fetal DV waveform (DV-no-A). However, many infants per monitoring protocol were delivered because of safety-net criteria, for maternal or other fetal indications, or after 32 weeks of gestation when the protocol was no longer applied. The objective of the present posthoc subanalysis was to investigate the indications for delivery in relation to 2-year outcome in infants delivered before 32 weeks to further refine management proposals. We included all 310 cases of the TRUFFLE study with known outcome at 2 years' corrected age and seven fetal deaths, excluding seven cases with inevitable perinatal death. Data were analyzed according to the allocated fetal monitoring strategy in combination with the indication for delivery. Overall, only 32% of liveborn infants were delivered according to the specified monitoring parameter for indication for delivery; 38% were delivered because of safety-net criteria, 15% for other fetal reasons and 15% for maternal reasons. In the CTG-STV group, 51% of infants were delivered because of reduced STV. In the DV-p95 group, 34% of infants were delivered because of abnormal DV and, in the DV-no-A group, only 10% of infants were delivered accordingly. The majority of infants in the DV groups were delivered for the safety-net criterion of spontaneous decelerations in FHR. Two-year intact survival was highest in the DV groups combined compared with the CTG-STV group (P = 0.05 for live births only, P = 0.21 including fetal death), with no difference between DV groups. A poorer outcome in the CTG-STV group was restricted to infants delivered because of FHR decelerations in the safety-net subgroup. Infants delivered because of maternal reasons had the highest birth weight and a non-significantly higher intact survival. In this subanalysis of infants delivered before 32 weeks, the majority were delivered for reasons other than the allocated monitoring strategy indication. Since, in the DV group, CTG-STV criteria were used as a safety net but in the CTG-STV group, no DV safety-net criteria were applied, we speculate that the slightly poorer outcome in the CTG-STV group might be explained by the absence of DV data. The optimal timing of delivery of fetuses with early intrauterine growth restriction may therefore be best determined by monitoring them longitudinally, with both DV and CTG monitoring. Copyright © 2016 ISUOG. Published by John Wiley & Sons Ltd.


Cell Density Counts of the Intestinal Intraepithelial Lymphocytes in the Celiac Patients.
Increased number of intestinal intraepithelial lymphocytes (IELs) is a key histological finding in the diagnosis of celiac disease (CD); however, the number of IELs in celiac patients and healthy subjects may vary from one region to another. Additionally, there are some seronegative celiac patients with a borderline histology. To determine the number of the CD3+ and CD8+ IELs T-cells in the celiac patients and healthy subjects (controls) in Isfahan. The duodenal biopsies were obtained from the celiac patients (n=15) and the controls (n=19). The total number of IELs/100 epithelial cells (ECs) were counted using the hematoxylin-eosin (H&E) staining method, and that of CD3+ and CD8+ IELs/100 ECs were counted using the immunohistochemistry (IHC) staining method. This study defined the upper normal limit for each variable as mean + 2SD. Accordingly, the upper normal limits of the total IELs, CD3+ IELs, and CD8+ IELs/100 ECs were calculated as 37 (95% confidence intervals, CI: 33-41), 22 (95% CI: 19-25) and 12 (95% CI: 10-14), respectively. In 3 clinically CD diagnoses, the total IELs counts/100 ECs were below the upper normal limit, and the histopathological and serologic assays were negative. Nevertheless, the CD8+ IELs T-cells counts/100 ECs showed borderline values. Interestingly, these patients responded to a gluten-free diet (GFD). The study findings suggest that in the clinically diagnosed celiac disease, IELs count/100 ECs below the upper normal limit as well as negative histopathological and serologic assays and the cell density counts of the CD8+ IELs T-cells/100 ECs could be a useful parameter in CD diagnosis and make a decision to put them on a GFD.


Unraveling the pathogenesis of Hoyeraal-Hreidarsson syndrome, a complex telomere biology disorder.
Hoyeraal-Hreidarsson (HH) syndrome is a multisystem genetic disorder characterized by very short telomeres and considered a clinically severe variant of dyskeratosis congenita. The main cause of mortality, usually in early childhood, is bone marrow failure. Mutations in several telomere biology genes have been reported to cause HH in about 60% of the HH patients, but the genetic defects in the rest of the patients are still unknown. Understanding the aetiology of HH and its diverse manifestations is challenging because of the complexity of telomere biology and the multiple telomeric and non-telomeric functions played by telomere-associated proteins in processes such as telomere replication, telomere protection, DNA damage response and ribosome and spliceosome assembly. Here we review the known clinical complications, molecular defects and germline mutations associated with HH, and elucidate possible mechanistic explanations and remaining questions in our understanding of the disease.


Wetland cover dynamics drive hemorrhagic disease patterns in white-tailed deer in the United States.
While vector-borne diseases are known to be particularly influenced by environmental factors, the impact of land-cover change on vector-borne wildlife disease patterns is poorly understood, largely due to the paucity of data on disease occurrence at extensive spatial and temporal scales. Widespread and rapid anthropogenic land-cover change, especially urbanization, has transformed the US landscape during the last century. Epizootic hemorrhagic disease virus and blue tongue virus, vectored by Culicoides biting midges, are two RNA viruses in the Orbivirus genus that cause severe hemorrhagic disease (HD) in white-tailed deer (Odocoileus virginianus). We examine the spatial dynamics of HD affecting white-tailed deer in the contiguous United States in two periods covering 1980 to 2007 in connection with land-cover change over the same time. Using spatial statistical modeling, wetland cover emerges as a critical driver of HD morbidity, whereas the drivers of mortality patterns are more complex. Increasing wetland cover is positively associated with HD morbidity, which is consistent with the ecologic requirements of the Culicoides vector. Wetland cover is inherently dynamic due to its importance to biodiversity and water quality as well as its utility for other purposes when drained. Accordingly this analysis helps in understanding the consequences of changing wetlands on vector-borne disease patterns, to identify disease hotspots in a large landscape, and to forecast the spatial spread of HD and related diseases.


Mitochondrial reactive oxygen species production mediates ursolic acid-induced mitochondrial uncoupling and glutathione redox cycling, with protection against oxidant injury in H9c2 cells.
Ursolic acid (UA), a natural pentacyclic triterpenoid carboxylic acid, is a ubiquitous compound widely distributed in many plants, fruits and medicinal herbs worldwide. A previous study in our laboratory has shown that UA can increase the mitochondrial ATP generation capacity (ATP-GC) and a glutathione-dependent antioxidant response, thereby protecting against oxidant injury in H9c2 cells in vitro and rat hearts ex vivo. However, the mechanism underlying the cellular protective effects induced by UA remains largely unknown. The present study has shown that pre-incubation with UA produces a transient increase in the mitochondrial membrane potential in H9c2 cells, which was accompanied by increases in mitochondrial reactive oxygen species (ROS) production. Studies using an antioxidant (dimethylthiourea) indicated that the suppression of mitochondrial ROS completely abrogated the UA-induced enhancement of mitochondrial uncoupling and glutathione reductase (GR)-mediated glutathione redox cycling, as well as protection against menadione cytotoxicity in H9c2 cells. Co-incubation with specific inhibitors of uncoupling proteins and GR almost completely prevented the cytoprotection afforded by UA against menadione-induced cytotoxicity in H9c2 cells. The results obtained so far suggest that UA-induced mitochondrial ROS production can elicit mitochondrial uncoupling and glutathione-dependent antioxidant responses, which offer cytoprotection against oxidant injury in H9c2 cells.


Eye movement responses to health messages on cigarette packages.
While the majority of the health messages on cigarette packages contain threatening health information, previous studies indicate that risk information can trigger defensive reactions, especially when the information is self-relevant (i.e., smokers). Providing coping information, information that provides help for quitting smoking, might increase attention to health messages instead of triggering defensive reactions. Eye-movement registration can detect attention preferences for different health education messages over a longer period of time during message exposure. In a randomized, experimental study with 23 smoking and 41 non-smoking student volunteers, eye-movements were recorded for sixteen self-created cigarette packages containing health texts that presented either high risk or coping information combined with a high threat or a low threat smoking-related photo. Results of the eye movement data showed that smokers tend to spend more time looking (i.e., more unique fixations and longer dwell time) at the coping information than at the high risk information irrespective of the content of the smoking-related photo. Non-smokers tend to spend more time looking at the high risk information than at the coping information when the information was presented in combination with a high threat smoking photo. When a low threat photo was presented, non-smokers paid more attention to the coping information than to the high risk information. Results for the smoking photos showed more attention allocation for low threat photos that were presented in combination with high risk information than for low threat photos in combination with coping information. No attention differences were found for the high threat photos. Non-smokers demonstrated an attention preference for high risk information as opposed to coping information, but only when text information was presented in combination with a high threat photo. For smokers, however, our findings suggest more attention allocation for coping information than for health risk information. This preference for coping information is not reflected in current health messages to motivate smokers to quit smoking. Coping information should be more frequently implemented in health message design to increase attention for these messages and thus contribute to effective persuasion.


Neural network classification of nerve activity recorded in a mixed nerve.
Whole-nerve cuff electrodes can be used to record electrical nerve activity in peripheral nerves and are suitable for chronic implantation in animals or humans. If the whole nerve innervates multiple target organs or muscles then the recorded activity will be the superposition of the activity of different nerve fibers innervating these organs. In certain cases it is desirable to monitor mixed nerve activity and to determine the origin (modality) of the recorded activity. A method using the autocorrelation function of recorded nerve activity and an artificial neural network was developed to classify the modality of nerve signals. The method works in cases where different end organs are innervated by nerve fibers having different diameter distributions. The electrical activity in the cat S1 sacral spinal root was recorded using a cuff electrode during the activation of cutaneous, bladder, and rectal mechanoreceptors. Using the classification method, 87.5% of nerve signals were correctly classified. This result demonstrates the effectiveness of the neural network classification method to determine the modality of the nerve activity arising from activation of different receptors.


Determining the Internal Validity of the Inventory of Reading Occupations: An Assessment Tool of Children's Reading Participation.
The Inventory of Reading Occupations (IRO) is an assessment tool of children's reading participation. In this study, we used Rasch methods to determine the internal validity of the IRO. Participants included 192 typical and struggling readers from kindergarten to third grade from five different states in the United States. We analyzed the fit of each of the items in the 17 reading categories, the test items in the three dimensions of reading participation, and the contexts of reading in the IRO. Analysis indicated that the IRO items support the Rasch model of unidimensionality. Analysis also indicated that 1 of the 30 test items can be revised to strengthen test validity. Moreover, the analysis also suggested that the IRO is more useful for first- to third-grade students. This study provides evidence of internal validity of a useful tool to assess children's reading participation.


Fever in travelers returning from tropical areas: prospective observational study of 613 cases hospitalised in Marseilles, France, 1999-2003.
Febrile travelers may pose a diagnostic challenge for Western physicians who are frequently involved in the assessment of these patients but unfamiliar with tropical diseases. Evaluation of this situation requires an understanding of the common etiologies, which are associated with the demographics of travelers and the destinations. We conducted a 5-year prospective observational study on the etiologies of fever in travelers returning from the tropics admitted to the infectious and tropical diseases unit of a university teaching hospital in Marseilles, France. A total of 613 patients were enrolled, including 364 migrants (59.4%), 126 travelers (20.6%), 37 visitors (6%), 24 expatriates (3.9%), and 62 patients (10.1%) who could not be classified. Malaria was the most common diagnosis (75.2%), with most cases (62%) acquired by migrants from the Comoros archipelago and who had traveled to these islands to visit friends and relatives. Agents of food-borne and water-borne infections (3.9%) and respiratory tract infections (3.4%) were also frequently identified as the cause of fever. Other infections included emerging diseases such as gnathostomiasis, hepatitis E infection and rickettsial diseases, as well as common infections or exotic diseases. Although we have identified here various causes of imported fever, 8.2% of the fevers remained unexplained. An improved approach to diagnosis may allow for the discovery of new diseases in travelers in the future.


The genetic basis of DOORS syndrome: an exome-sequencing study.
Deafness, onychodystrophy, osteodystrophy, mental retardation, and seizures (DOORS) syndrome is a rare autosomal recessive disorder of unknown cause. We aimed to identify the genetic basis of this syndrome by sequencing most coding exons in affected individuals. Through a search of available case studies and communication with collaborators, we identified families that included at least one individual with at least three of the five main features of the DOORS syndrome: deafness, onychodystrophy, osteodystrophy, intellectual disability, and seizures. Participants were recruited from 26 centres in 17 countries. Families described in this study were enrolled between Dec 1, 2010, and March 1, 2013. Collaborating physicians enrolling participants obtained clinical information and DNA samples from the affected child and both parents if possible. We did whole-exome sequencing in affected individuals as they were enrolled, until we identified a candidate gene, and Sanger sequencing to confirm mutations. We did expression studies in human fibroblasts from one individual by real-time PCR and western blot analysis, and in mouse tissues by immunohistochemistry and real-time PCR. 26 families were included in the study. We did exome sequencing in the first 17 enrolled families; we screened for TBC1D24 by Sanger sequencing in subsequent families. We identified TBC1D24 mutations in 11 individuals from nine families (by exome sequencing in seven families, and Sanger sequencing in two families). 18 families had individuals with all five main features of DOORS syndrome, and TBC1D24 mutations were identified in half of these families. The seizure types in individuals with TBC1D24 mutations included generalised tonic-clonic, complex partial, focal clonic, and infantile spasms. Of the 18 individuals with DOORS syndrome from 17 families without TBC1D24 mutations, eight did not have seizures and three did not have deafness. In expression studies, some mutations abrogated TBC1D24 mRNA stability. We also detected Tbc1d24 expression in mouse phalangeal chondrocytes and calvaria, which suggests a role of TBC1D24 in skeletogenesis. Our findings suggest that mutations in TBC1D24 seem to be an important cause of DOORS syndrome and can cause diverse phenotypes. Thus, individuals with DOORS syndrome without deafness and seizures but with the other features should still be screened for TBC1D24 mutations. More information is needed to understand the cellular roles of TBC1D24 and identify the genes responsible for DOORS phenotypes in individuals who do not have a mutation in TBC1D24. US National Institutes of Health, the CIHR (Canada), the NIHR (UK), the Wellcome Trust, the Henry Smith Charity, and Action Medical Research.


Wildfire-Induced CO Plume Observations From NAST-I During the FIREX-AQ Field Campaign.
The fire influence on regional to global environments and air quality (FIREX-AQ) field campaign was conducted during August 2019 to investigate the impact of wildfire and biomass smoke on air quality and weather in the continental United States. One of the campaign's scientific objectives was to estimate the composition of emissions from wildfires. Ultraspectrally resolved infrared radiance measurements from aircraft and/or satellite observations contain information on tropospheric carbon monoxide (CO) as well as other trace species present in fire emissions. A methodology for retrieving tropospheric CO from such remotely sensed spectral data has been developed for the National Airborne Sounder Testbed-Interferometer (NAST-I) and is applied herein. Retrievals based on NAST-I measurements are used to demonstrate CO retrieval capability and characterize fire emissions. NAST-I remotely sensed CO from ER-2 flights are evaluated with concurrent in situ measurements from the differential absorption carbon monoxide measurements flown on the NASA DC-8 aircraft. Enhanced CO emissions along with plume evolution and transport from the fire ground site locations were captured by moderate vertical and high horizontal resolution observations obtained from the NAST-I IR spectrometer; these were intercompared and verified by the cloud physics lidar and the enhanced MODIS airborne simulator also hosted on the NASA ER-2 aircraft. This study will be beneficial to the science community for studying wildfire-related topics and understanding similar remotely sensed observations from satellites, along with helping to address the broader FIREX-AQ experiment objectives of investigating the impact of fires on air quality and climate.


Encapsulation of angiogenic monocytes using bio-spraying technology.
Therapeutic neovascularisation using angiogenic cells has been hampered by the loss of cells from the target tissue. Encapsulation of these cells within a semi-permeable membrane could improve their retention within the ischaemic tissue without affecting the excretion of the angiogenic growth factors produced. Bio-spraying is a novel cell-handling technique that does not adversely affect cell viability. We used this technique to encapsulate human peripheral blood monocytes and found that cell viability, cell phenotype and functional downstream angiogenic signalling were preserved. Encapsulation of monocytes with macrophage-colony stimulating factor resulted in increased vascular-endothelial growth factor production and enhanced angiogenic function. Bio-spraying/encapsulation has the potential to enhance the efficacy of current angiogenic cell therapy strategies and merits further investigation.