Degradation of antiflammin 2 under acidic conditions.
Antiflammin 2 (HDMNKVLDL, AF2) is a synthetic peptide derived from the region of highest sequence similarity of lipocortin I and uteroglobin, and is a potent antiinflammatory agent without any known side effects of corticosteroids. The antiinflammatory activity of AF2 has been demonstrated, but is not reproducible between laboratories. It has been suggested that the chemical instability of this peptide is responsible for the loss of activity. The degradation of AF2 in aqueous solutions at a pH range of 3 to 10 has been reported. In this study, the degradation of AF2 at acidic pHs was monitored by reversed-phase HPLC. The reactions were studied as functions of buffer concentration and temperature. The rates of loss of AF2 followed apparent pseudo-first-order kinetics. Several products were isolated and identified by fast atom bombardment mass spectroscopy and tandem mass spectroscopy, and were the result of C- and N-terminus hydrolyses of aspartyl peptide bonds in AF2. The peptide bonds at C-termini of the aspartyl residues were most susceptible to hydrolysis, resulting in the formation of major degradation products, HDMNKVLD, MNKVLDL, and MNKVLD. The minor products from the N-terminus hydrolysis were HDMNKVL and MNKVL and formed at much slower rates.


Effects of a long-term lifestyle modification programme on peripheral neuropathy in overweight or obese adults with type 2 diabetes: the Look AHEAD study.
The aim of this study was to evaluate the effects on diabetic peripheral neuropathy (DPN) of a long-term intensive lifestyle intervention (ILI) programme designed to achieve and maintain weight loss. Beginning in 2001, a total of 5145 overweight or obese people with type 2 diabetes, aged 45-76 years, participating in the multicentre Look AHEAD (Action for Health in Diabetes) study were randomised to ILI (n = 2570) or to a diabetes support and education (DSE) control group (n = 2575) using a web-based management system at the study coordinating centre at Wake Forest School of Medicine (Winston-Salem, NC, USA). Randomisation was stratified by clinical centre and was not revealed to the clinical staff responsible for obtaining data on study outcomes. Because of the nature of the study, patients and the local centre interventionists were not blinded to the study group assignments. In addition, the coordinating centre staff members responsible for data management and statistical analyses were not blinded to the study group assignments. The interventions were terminated in September 2012, 9-11 years after randomisation, but both groups continued to be followed for both primary and secondary outcomes. Neuropathy evaluations included the Michigan Neuropathy Screening Instrument (MNSI) questionnaire completed at baseline in 5145 participants (ILI n = 2570, DSE n = 2575) and repeated annually thereafter and the MNSI physical examination and light touch sensation testing conducted in 3775 participants (ILI n = 1905, DSE n = 1870) 1-2.3 years after discontinuation of the intervention. At baseline, the MNSI questionnaire scores were 1.9 ± 0.04 and 1.8 ± 0.04 in the ILI and DSE groups, respectively (difference not statistically significant). After 1 year, when weight loss was maximal in the ILI group (8.6 ± 6.9%) compared with DSE (0.7 ± 4.8%), the respective MNSI scores were 1.7 ± 0.04 and 2.0 ± 0.04 (p ≤ 0.001). Subsequently, the scores increased gradually in both groups, but remained significantly lower in the ILI group for the first 3 years and at the end of follow-up. In both groups, there was a significant association between changes in the MNSI scores and changes in body weight, HbA1c and serum lipids. There were no significant between-group differences in the proportions of participants with MNSI physical examination scores ≥2.5, considered to be indicative of diabetic neuropathy. The light touch sensation measured separately in either the right or left big toes (halluces) did not differ between ILI and DSE, but when the data were combined for both toes, light touch was better preserved in the ILI group. ILI resulted in a significant decrease in questionnaire-based DPN, which was associated with the magnitude of weight loss. In both the ILI and DSE groups, changes in the MNSI score were also related to changes in HbA1c and lipids. There were no significant effects of ILI on physical examination measures of DPN conducted 1-2.3 years after termination of the active intervention, except for light touch sensation, which was significantly better in the ILI group when measurements were combined for both toes. However, a potential limiting factor to the interpretation of the physical examination data is that no baseline studies are available for comparison. ClinicalTrials.gov NCT00017953. This work was funded by the National Institutes of Health through cooperative agreements with the National Institute of Diabetes and Digestive and Kidney Diseases.


Washing our hands of the congenital cytomegalovirus disease epidemic.
Each year in the United States, an estimated 40,000 children are born with congenital cytomegalovirus (CMV) infection, causing an estimated 400 deaths and leaving approximately 8000 children with permanent disabilities such as hearing or vision loss, or mental retardation. More children are affected by serious CMV-related disabilities than by several better-known childhood maladies, including Down syndrome, fetal alcohol syndrome, and spina bifida. Congenital CMV is a prime target for prevention not only because of its substantial disease burden but also because the biology and epidemiology of CMV suggest that there are ways to reduce viral transmission. Because exposure to the saliva or urine of young children is a major cause of CMV infection among pregnant women, it is likely that good personal hygiene, especially hand-washing, can reduce the risk of CMV acquisition. Experts agree that such measures are likely to be efficacious (i.e., they will work if consistently followed) and the American College of Obstetricians and Gynecologists recommends that physicians counsel pregnant women about preventing CMV acquisition through careful attention to hygiene. However, because of concerns about effectiveness (i.e., Will women consistently follow hygienic practices as the result of interventions?), the medical and public health communities appear reluctant to embrace primary CMV prevention via improved hygienic practices, and educational interventions are rare. Current data on the effectiveness of such measures in preventing CMV infection are promising, but limited. There is strong evidence, however, that educational interventions can prevent other infectious diseases with similar transmission modes, suggesting that effective interventions can also be found for CMV. Until a CMV vaccine becomes available, effective educational interventions are needed to inform women about congenital CMV prevention. Perhaps no single cause of birth defects and developmental disabilities in the United States currently provides greater opportunity for improved outcomes in more children than congenital CMV. Given the present state of knowledge, women deserve to be informed about how they can reduce their risk of CMV infection during pregnancy, and trials are needed to identify effective educational interventions.


Investigation of mean platelet volume in patients with type 2 diabetes mellitus and in subjects with impaired fasting glucose: a cost-effective tool in primary health care?
The aim of this study was to compare mean platelet volume (MPV) in patients with type 2 diabetes mellitus (T2DM), in subjects with impaired fasting glucose (IFG), and in non-diabetic controls. A total of 201 adults with T2DM and 201 subjects with IFG from the Family Medicine out-patient clinic as well as 201 healthy controls were included in the study. We measured blood fasting glucose, complete blood count and LDL-cholesterol and compared the results between the groups enrolled. In the patients with diabetes and subjects with IFG, MPV was significantly higher (10.66 ± 0.94 fL and 10.49 ± 0.96 fL, respectively ) as compared to the non-diabetic group (10.04 ± 1.01 fL) (p = 0.000). Among the diabetic subjects, a positive statistical Pearson correlation was seen between MPV and HbA1c levels (r = 0.357; p = 0.000) and fasting blood glucose (FBG) levels (r = 0.306; p = 0.000). The mean MPV in patients having HbA1C < 7.5% was 10.17 ± 0.83 fL and significantly lower than that of patients with HbA1c ≥ 7.5% (10.80 ± 0.92 fL) (p = 0.001). MPV could be used as a simple and cost-effective tool to monitor the progression and control of T2DM and thereby in preventing vascular events in primary health care.


Polyclonal Expansion of NKG2C(+) NK Cells in TAP-Deficient Patients.
Adaptive natural killer (NK) cell responses to human cytomegalovirus infection are characterized by the expansion of NKG2C(+) NK cells expressing self-specific inhibitory killer-cell immunoglobulin-like receptors (KIRs). Here, we set out to study the HLA class I dependency of such NKG2C(+) NK cell expansions. We demonstrate the expansion of NKG2C(+) NK cells in patients with transporter associated with antigen presentation (TAP) deficiency, who express less than 10% of normal HLA class I levels. In contrast to normal individuals, expanded NKG2C(+) NK cell populations in TAP-deficient patients display a polyclonal KIR profile and remain hyporesponsive to HLA class I-negative target cells. Nonetheless, agonistic stimulation of NKG2C on NK cells from TAP-deficient patients yielded significant responses in terms of degranulation and cytokine production. Thus, while interactions with self-HLA class I molecules likely shape the KIR repertoire of expanding NKG2C(+) NK cells during adaptive NK cell responses in normal individuals, they are not a prerequisite for NKG2C(+) NK cell expansions to occur. The emergence of NKG2C-responsive adaptive NK cells in TAP-deficient patients may contribute to antiviral immunity and potentially explain these patients' low incidence of severe viral infections.


Are birthweight and postnatal weight gain in childhood associated with blood pressure in early adolescence? Results from a Ugandan birth cohort.
In Africa, where low birthweight (LBW), malnutrition and high blood pressure (BP) are prevalent, the relationships between birthweight (BW), weight gain and BP later in life remain uncertain. We examined the effects of early life growth on BP among Ugandan adolescents. Data were collected prenatally from women and their offspring were followed from birth, with BP measured following standard protocols in early adolescence. Weight-for-age Z-scores (WAZ) were computed using World Health Organization references. Linear regression was used to relate BW, and changes in WAZ between birth and 5 years, to adolescents' BP, adjusting for confounders. Among 2345 live offspring, BP was measured in 1119 (47.7%) adolescents, with mean systolic BP 105.9 mmHg and mean diastolic BP 65.2 mmHg. There was little evidence of association between BW and systolic [regression coefficient β = 0.14, 95% confidence interval (CI) (-1.00, 1.27)] or diastolic [β = 0.43, 95% CI (-0.57, 1.43)] BP. Accelerated weight gain between birth and 5 years was associated with increased BP: systolic β = 1.17, 95% CI (0.69, 1.66) and diastolic β = 1.03, 95% CI (0.59, 1.47). Between birth and 6 months of age, effects of accelerated weight gain on adolescent BP were strongest among the LBW (both premature and small-for-gestational-age) children [BW < 2.5 kg: β = 2.64, 95% CI (0.91, 4.37), BW≥2.5 kg: β = 0.58, 95% CI (0.01, 1.14), interaction P-value = 0.024]. Findings from this large tropical birth cohort in Uganda suggest that postnatal weight gain rather than BW is important in the developmental programming of BP, with fast-growing LBW children at particular risk. Efforts to control BP should adopt a life course approach.


Antibacterial, antifungal, phytotoxic, antioxidant and hemagglutination activities of organic fractions of Arisaema tortuosum.
In the current study, the antimicrobial, phytotoxic, haemagglutination and antioxidant potential of crude methanolic extract (Crd. MeOH Ext.) and four organic fractions of Arisaema tortuosum was investigated. All fractions have been screened for antimicrobial properties against eight bacterial pathogens and six fungal pathogens using agar well diffusion and tube dilution method, respectively. Furthermore, the organic fractions were also screened for its phytotoxicity against Lemna minor. Haemagglutination was performed against all human blood groups while free radical scavenging activity was performed to investigate the antioxidant potential of A. tortuosum. Results obtained for antibacterial activity exhibited various degree of zone of inhibition and significant activity was observed for Pseudomonas aeruginosa (27.16±0.60) followed by Bacillus cereus (18.55±0.69) for Crd. MeOH Ext. and chloroform (CHCl3) fraction, respectively while some strains showed resistant at same concentration. Similarly, non-significant antifungal activity was observed for the plant extracts. However, the highest activity among the strains was observed for Alternaria alternata (22±1.24%) and Aspergillus niger (20±1.00%) for ethyl acetate (EtOAc) fraction and Crd. MeOH Ext., respectively. The plant extracts showed good phytotoxic activity with 77.06% inhibition for n-hexane fraction at 1000µg/mL. The result of Nitric Oxide (NO) reducing assay revealed that the plant has less antioxidant activity with 46.06% inhibition for CHCl(3) fraction at 900μg/mL. For haemagglutination assay, the result displayed no agglutination in all the testing concentration. Based on the current results, it can be concluded that A. tortuosum has significant antimicrobial and moderate phytotoxic potential and therefore can leads to antibiotics and herbicide production.


Developing a Complex Educational-Behavioural Intervention: The TREAT Intervention for Patients with Atrial Fibrillation.
This article describes the theoretical and pragmatic development of a patient-centred intervention for patients with atrial fibrillation (AF). Theoretical models (Common Sense Model, Necessity-Concerns Framework), clinical frameworks, and AF patient feedback contributed to the design of a one-off hour-long behaviour-change intervention package. Intervention materials consisted of a DVD, educational booklet, diary and worksheet, which were patient-centred and easy to administer. The intervention was evaluated within a randomised controlled trial. Several "active theoretical ingredients" were identified (for e.g., where patients believed their medication was less harmful they spent more time within the therapeutic range (TTR), with general harm scores predicting TTR at 6 months). Allowing for social comparison and adopting behaviour change techniques enabled accurate patient understanding of their condition and medication. The process of developing the intervention using theory-derived content and evaluation tools allowed a greater understanding of the mechanisms by which this intervention was successful. Alleviating concerns about treatment medication by educating patients can help to improve adherence. This process of intervention development could be adopted for a range of chronic illnesses and treatments. Critical elements should include the use of: (1) clinical guidelines; (2) appropriate theoretical models; (3) patient input; and (4) appropriate evaluation tools.


Position-dependent motif characterization using non-negative matrix factorization.
Cis-acting regulatory elements are frequently constrained by both sequence content and positioning relative to a functional site, such as a splice or polyadenylation site. We describe an approach to regulatory motif analysis based on non-negative matrix factorization (NMF). Whereas existing pattern recognition algorithms commonly focus primarily on sequence content, our method simultaneously characterizes both positioning and sequence content of putative motifs. Tests on artificially generated sequences show that NMF can faithfully reproduce both positioning and content of test motifs. We show how the variation of the residual sum of squares can be used to give a robust estimate of the number of motifs or patterns in a sequence set. Our analysis distinguishes multiple motifs with significant overlap in sequence content and/or positioning. Finally, we demonstrate the use of the NMF approach through characterization of biologically interesting datasets. Specifically, an analysis of mRNA 3'-processing (cleavage and polyadenylation) sites from a broad range of higher eukaryotes reveals a conserved core pattern of three elements.


A simplified roentgen stereophotogrammetric method. Analysis of small movements between the prosthetic stem and the femur after total hip replacement.
A simplified roentgen stereophotogrammetric method is described. It is based on the use of a 50 mm thick reference plate consisting of a carbon-fibre-reinforced polyester box. The patient is placed directly on this box, which makes the methods less cumbersome and more suitable for routine use. The method has been tested in a model experiment designed for detecting small movements between femur and prosthesis at an early stage after total hip replacement. The head and two hemispheres on the prosthesis and three small tantalum balls inserted in the femur serve as reference points. The model experiment now reported shows that the method has acceptable precision.


Chest Fat Quantification via CT Based on Standardized Anatomy Space in Adult Lung Transplant Candidates.
Overweight and underweight conditions are considered relative contraindications to lung transplantation due to their association with excess mortality. Yet, recent work suggests that body mass index (BMI) does not accurately reflect adipose tissue mass in adults with advanced lung diseases. Alternative and more accurate measures of adiposity are needed. Chest fat estimation by routine computed tomography (CT) imaging may therefore be important for identifying high-risk lung transplant candidates. In this paper, an approach to chest fat quantification and quality assessment based on a recently formulated concept of standardized anatomic space (SAS) is presented. The goal of the paper is to seek answers to several key questions related to chest fat quantity and quality assessment based on a single slice CT (whether in the chest, abdomen, or thigh) versus a volumetric CT, which have not been addressed in the literature. Unenhanced chest CT image data sets from 40 adult lung transplant candidates (age 58 ± 12 yrs and BMI 26.4 ± 4.3 kg/m2), 16 with chronic obstructive pulmonary disease (COPD), 16 with idiopathic pulmonary fibrosis (IPF), and the remainder with other conditions were analyzed together with a single slice acquired for each patient at the L5 vertebral level and mid-thigh level. The thoracic body region and the interface between subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) in the chest were consistently defined in all patients and delineated using Live Wire tools. The SAT and VAT components of chest were then segmented guided by this interface. The SAS approach was used to identify the corresponding anatomic slices in each chest CT study, and SAT and VAT areas in each slice as well as their whole volumes were quantified. Similarly, the SAT and VAT components were segmented in the abdomen and thigh slices. Key parameters of the attenuation (Hounsfield unit (HU) distributions) were determined from each chest slice and from the whole chest volume separately for SAT and VAT components. The same parameters were also computed from the single abdominal and thigh slices. The ability of the slice at each anatomic location in the chest (and abdomen and thigh) to act as a marker of the measures derived from the whole chest volume was assessed via Pearson correlation coefficient (PCC) analysis. The SAS approach correctly identified slice locations in different subjects in terms of vertebral levels. PCC between chest fat volume and chest slice fat area was maximal at the T8 level for SAT (0.97) and at the T7 level for VAT (0.86), and was modest between chest fat volume and abdominal slice fat area for SAT and VAT (0.73 and 0.75, respectively). However, correlation was weak for chest fat volume and thigh slice fat area for SAT and VAT (0.52 and 0.37, respectively), and for chest fat volume for SAT and VAT and BMI (0.65 and 0.28, respectively). These same single slice locations with maximal PCC were found for SAT and VAT within both COPD and IPF groups. Most of the attenuation properties derived from the whole chest volume and single best chest slice for VAT (but not for SAT) were significantly different between COPD and IPF groups. This study demonstrates a new way of optimally selecting slices whose measurements may be used as markers of similar measurements made on the whole chest volume. The results suggest that one or two slices imaged at T7 and T8 vertebral levels may be enough to estimate reliably the total SAT and VAT components of chest fat and the quality of chest fat as determined by attenuation distributions in the entire chest volume.


A Qualitative Exploration of the Economic and Social Effects of Microcredit among People Living with HIV/AIDS in Uganda.
HIV medical care, including antiretroviral therapy (ART), is often successful in restoring physical health and functioning. But in developing countries, HIV medical care is often insufficient to achieve social and economic health, and hence innovative economic support programs are much needed. We conducted semistructured interviews with 30 adults receiving ART and microcredit loans operated by Uganda Cares. Using content analysis, we explored the impact of the microcredit loans on the economic, social, and psychological well-being of respondents. Most respondents indicated that the microcredit loans played a positive role in their lives, helped them to keep their children in school and sustain their families, and improved their self-esteem and status in the community. In addition, we also found significant positive knowledge spill-over and network effects in the program with regard to business management and support. However, more than half of the participants indicated experiencing repayment problems either personally or with other group members due to unexpected emergencies and sickness. These findings highlight that microcredit programs have the potential of being an economic support system for HIV clients trying to reestablish their livelihoods, especially in resource-constrained settings, though more research is needed to determine the overall economic viability of such programs.


COMMUNITY MEMBERS' PERCEPTIONS OF MASS DRUG ADMINISTRATION FOR CONTROL OF LYMPHATIC FILARIASIS IN RURAL AND URBAN TANZANIA.
Lymphatic filariasis is one of several neglected tropical diseases with severely disabling and stigmatizing manifestations that are referred to as 'neglected diseases of poverty'. It is a mosquito-borne disease found endemically and exclusively in low-income contexts where, concomitantly, general public health care is often deeply troubled and fails to meet the basic health needs of impoverished populations. This presents particular challenges for the implementation of mass drug administration (MDA), which currently is the principal means of control and eventual elimination. Several MDA programmes face the dilemma that they are unable to attain and maintain the required drug coverage across target groups. In recognition of this, a qualitative study was conducted in the Morogoro and Lindi regions of Tanzania to gain an understanding of community experiences with, and perceptions of, the MDA campaign implemented in 2011 by the National Lymphatic Filariasis Elimination Programme. The study revealed a wide variation of perceptions and experiences regarding the aim, rationale and justification of MDA. There were positive sentiments about the usefulness of the drugs, but many study participants were sceptical about the manner in which MDA is implemented. People were particularly disappointed with the limited attempts by implementers to share information and mobilize residents. In addition, negative sentiments towards MDA for lymphatic filariasis reflected a general feeling of desertion and marginalization by the health care system and political authorities. However, the results suggest that if the communities are brought on board with genuine respect for their integrity and informed self-determination, there is scope for major improvements in community support for MDA-based control activities.


Evaluation of Quality of Life and Satisfaction of Patients with Neuropathic Pain and Breakthrough Pain: Economic Impact Based on Quality of Life.
The study objective was to assess the quality of life and satisfaction with treatment of patients with chronic neuropathic pain (CNP) who experience breakthrough pain (BTP) and to assess its economic impact. Cross-sectional observational study. Fifteen pain units from Spanish hospitals completed the study. A total of 124 patients with adequately controlled CNP who experienced BTP were enrolled into the study. No interventions were required. Quality of life was assessed using the SF12 v2 questionnaire, the results of which were used to calculate the estimated costs per patient and month and the SF-6D Health Utility Index. Patient satisfaction with treatment received for CNP and for BTP was assessed using a 10-point visual analogue scale. Other associated symptoms were analyzed using the ESAS (Edmonton Symptom Assessment System). Patients had a mean age of 60.2 years (95% CI 58.4-63.3), and 46.8% (58) were males. 18.9% (23) experienced their first episode of BTP. A severe impairment of the physical component of SF12v2 was noted, with 94% of patients below the mean score of the population, while 88% had values lower than normal for the mental component. Mean cost per patient and month was $679 and was significantly greater in males ($763 versus $606), 4.96 times greater than in healthy population, and approximately double the cost of patients with CNP in Spain. Occurrence of BTP in patients with CNP causes a substantial increase in healthcare costs which is significantly greater in older males.


Genome analyses reveal the hybrid origin of the staple crop white Guinea yam (Dioscorea rotundata).
White Guinea yam (Dioscorea rotundata) is an important staple tuber crop in West Africa. However, its origin remains unclear. In this study, we resequenced 336 accessions of white Guinea yam and compared them with the sequences of wild Dioscorea species using an improved reference genome sequence of D. rotundata In contrast to a previous study suggesting that D. rotundata originated from a subgroup of Dioscorea praehensilis, our results suggest a hybrid origin of white Guinea yam from crosses between the wild rainforest species D. praehensilis and the savannah-adapted species Dioscorea abyssinica We identified a greater genomic contribution from D. abyssinica in the sex chromosome of Guinea yam and extensive introgression around the SWEETIE gene. Our findings point to a complex domestication scenario for Guinea yam and highlight the importance of wild species as gene donors for improving this crop through molecular breeding.


Controlling Posture and Vergence Eye Movements in Quiet Stance: Effects of Thin Plantar Inserts.
The purpose of this study was to assess properties of vergence and saccade eye movements as well as posture in quiet stance, and the effects of thin plantar inserts upon postural and oculomotor control. The performances of 36 young healthy subjects were recorded by a force platform and an eye tracker in three testing conditions: without plantar stimulation, with a 3 millimetre-thick plantar insert, either a Medial or a Lateral Arch Support (MAS/LAS). The results showed a decrease of the Surface and Variance of Speed and a more posterior position of the CoP with either stimulation compared with the control condition. The fractal analysis showed a decrease with MAS. Wavelet analysis in the time-frequency domain revealed an increase in the Cancelling Time of the low frequency band with MAS. These results suggest a better stability for a lower energy cost. Concerning eye movements, the inserts influenced only vergence (not saccades): MAS caused an increase of the phasic amplitude of divergence, and conversely a decrease of the tonic amplitude. In contrast, LAS caused an increase of the tonic amplitude of convergence. Thus, MAS renders divergence less visually driven, while LAS renders convergence more visually driven. We conclude that the CNS uses the podal signal for both postural and vergence control via specific mechanisms. Plantar inserts have an influence upon posture and vergence movements in a different way according to the part of the foot sole being stimulated. These results can be useful to clinicians interested in foot or eye.


Dissection of a Ciona regulatory element reveals complexity of cross-species enhancer activity.
Vertebrate genomes share numerous conserved non-coding elements, many of which function as enhancer elements and are hypothesised to be under evolutionary constraint due to a need to be bound by combinations of sequence-specific transcription factors. In contrast, few such conserved elements can be detected between vertebrates and their closest invertebrate relatives. Despite this lack of sequence identity, cross-species transgenesis has identified some cases where non-coding DNA from invertebrates drives reporter gene expression in transgenic vertebrates in patterns reminiscent of the expression of vertebrate orthologues. Such instances are presumed to reflect the presence of conserved suites of binding sites in the regulatory regions of invertebrate and vertebrate orthologues, such that both regulatory elements can correctly interpret the trans-activating environment. Shuffling of binding sites has been suggested to lie behind loss of sequence conservation; however this has not been experimentally tested. Here we examine the underlying basis of enhancer activity for the Ciona intestinalis βγ-crystallin gene, which drives expression in the lens of transgenic vertebrates despite the Ciona lineage predating the evolution of the lens. We construct an interactive gene regulatory network (GRN) for vertebrate lens development, allowing network interactions to be robustly catalogued and conserved network components and features to be identified. We show that a small number of binding motifs are necessary for Ciona βγ-crystallin expression, and narrow down the likely factors that bind to these motifs. Several of these overlap with the conserved core of the vertebrate lens GRN, implicating these sites in cross species function. However when we test these motifs in a transgenic vertebrate they prove to be dispensable for reporter expression in the lens. These results show that current models depicting cross species enhancer function as dependent on conserved binding sites can be overly simplistic, with sound evolutionary inference requiring detailed dissection of underlying mechanisms.


New fluorescent method (PLT-F) on Sysmex XN2000 hematology analyzer achieved higher accuracy in low platelet counting.
In thrombocytopenia, high accuracy and precision of low platelet count is essential for appropriate decisions. The recently introduced Sysmex XN2000 analyzer (Sysmex, Kobe, Japan) offers 3 methods for platelet counting: impedance (PLT-I), optical (PLT-O), and a new fluorescence method (PLT-F). The precision of the PLT-F method in blood samples with platelet counts less than 50 × 10(3)/μL (50 × 10(9)/L) was investigated and compared with the ICSH CD61-ImmunoPLT reference method. For comparison, PLT-I and PLT-O were determined on the Sysmex XN2000 and Sysmex XE2100 analyzer. Blood samples with platelet counts less than 50 × 10(3)/μL (50 × 10(9)/L) (n = 37) were analyzed on the Sysmex XN2000 and XE2100 analyzers. The CD61-ImmunoPLT method was performed on a Beckman Coulter FC-500 flow cytometer (Miami, FL). At a platelet count of 20 × 10(3)/μL (20 × 10(9)/L), reproducibility for PLT-I, PLT-O, and PLT-F on the XN2000 demonstrated coefficients of variation of 9.3%, 8.5%, and 3.0%, respectively. Correlation between PLT-O on the XN2000 and XE2100 yielded an r value of more than 0.977. Linear regression analysis between the PLT-F and CD61-ImmunoPLT methods resulted in a PLT-F of 0.71*CD61 - 0.8 (r = 0.988). Linear regression between PLT-F and PLT-O on the XN2000 resulted in a PLT-F of 1.05*PLT-O - 2 (r = 0.975), and using the transfusion threshold of 20 × 10(9)/L platelets resulted in a PLT-F of 0.90*PLT-O - 0.4 (r = 0.956). The new PLT-F method demonstrated excellent results for reproducibility in samples with platelet counts less than 50 × 10(9)/L. PLT-F could be helpful in making better decisions for platelet transfusions.


Red blood cells of sickle cell disease patients exhibit abnormally high abundance of N-methyl D-aspartate receptors mediating excessive calcium uptake.
Recently we showed that N-methyl D-aspartate receptors (NMDARs) are expressed in erythroid precursors (EPCs) and present in the circulating red blood cells (RBCs) of healthy humans, regulating intracellular Ca(2+) in these cells. This study focuses on investigating the possible role of NMDARs in abnormally high Ca(2+) permeability in the RBCs of patients with sickle cell disease (SCD). Protein levels of the NMDAR subunits in the EPCs of SCD patients did not differ from those in EPCs of healthy humans. However, the number and activity of the NMDARs in circulating SCD-RBCs was substantially up-regulated, being particularly high during haemolytic crises. The number of active NMDARs correlated negatively with haematocrit and haemoglobin levels in the blood of SCD patients. Calcium uptake via these non-selective cation channels was induced by RBC treatment with glycine, glutamate and homocysteine and was facilitated by de-oxygenation of SCD-RBCs. Oxidative stress and RBC dehydration followed receptor stimulation and Ca(2+) uptake. Inhibition of the NMDARs with an antagonist memantine caused re-hydration and largely prevented hypoxia-induced sickling. The EPCs of SCD patients showed higher tolerance to memantine than those of healthy subjects. Consequently, NMDARs in the RBCs of SCD patients appear to be an attractive target for pharmacological intervention.


The cooperative University of Iowa / Iowa State University MPH program.
Public health is an important component of veterinary medicine. In the last 10 years, there has been growing recognition of the need to increase the number of veterinarians trained in public health. The Center for Food Security and Public Health (CFSPH) at Iowa State University (ISU), College of Veterinary Medicine, received a grant from the Centers for Disease Control and Prevention (CDC) to support veterinarians working at CFSPH while pursuing the Master of Public Health degree. CFSPH and ISU administrators worked with the University of Iowa (UI) College of Public Health to establish three cooperative programs for veterinarians to earn the MPH degree. This article describes how these programs were developed and how they operate. (1) Between 2002 and 2005, CFSPH used funds provided by the CDC to support 15 veterinarians as they worked for CFSPH and toward the MPH degree. As the program grew, distance-education methods such as the Internet, Polycom videoconferencing, and the Iowa Communications Network (ICN) were incorporated. (2) A concurrent DVM/MPH degree is now offered; students can complete both degrees in four years. As of January 2008, three students have received their DVM and MPH degrees and 16 students are enrolled in the program. (3) In June 2007, the UI and ISU launched a distance MPH program for veterinarians working in private practice, industry, and government. Eight veterinarians are participating in the program, which includes two two-week, in-person summer sessions, with the remainder of the coursework taken at a distance via the Internet.


HIV-Specific B Cell Frequency Correlates with Neutralization Breadth in Patients Naturally Controlling HIV-Infection.
HIV-specific broadly neutralizing antibodies (bnAbs) have been isolated from patients with high viremia but also from HIV controllers that repress HIV-1 replication. In these elite controllers (ECs), multiple parameters contribute to viral suppression, including genetic factors and immune responses. Defining the immune correlates associated with the generation of bnAbs may help in designing efficient immunotherapies. In this study, in ECs either positive or negative for the HLA-B*57 protective allele, in treated HIV-infected and HIV-negative individuals, we characterized memory B cell compartments and HIV-specific memory B cells responses using flow cytometry and ELISPOT. ECs preserved their memory B cell compartments and in contrast to treated patients, maintained detectable HIV-specific memory B cell responses. All ECs presented IgG1+ HIV-specific memory B cells but some individuals also preserved IgG2+ or IgG3+ responses. Importantly, we also analyzed the capacity of sera from ECs to neutralize a panel of HIV strains including transmitted/founder virus. 29% and 21% of HLA-B*57+ and HLA-B*57- ECs, respectively, neutralized at least 40% of the viral strains tested. Remarkably, in HLA-B*57+ ECs the frequency of HIV-Env-specific memory B cells correlated positively with the neutralization breadth suggesting that preservation of HIV-specific memory B cells might contribute to the neutralizing responses in these patients.


Fate of the initial follicle pool: empirical and mathematical evidence supporting its sufficiency for adult fertility.
The importance of the initial follicle pool in fertility in female adult mammals has recently been debated. Utilizing a mathematical model of the dynamics of follicle progression (primordial to primary to secondary), we examined whether the initial follicle pool is sufficient for adult fertility through reproductive senescence in CD1 mice. Follicles in each stage were counted from postnatal day 6 through 12 months and data were fit to a series of first-order differential equations representing two mechanisms: an initial pool of primordial follicles as the only follicle source (fixed pool model), or an initial primordial follicle pool supplemented by germline stem cells (stem cell model). The fixed pool model fit the experimental data, accurately representing the maximum observed primary follicle number reached by 4-6 months of age. Although no germline stem cells could be identified by SSEA-1 immunostaining, the stem cell model was tested using a range of de novo primordial follicle production rates. The stem cell model failed to describe the observed decreases in follicles over time and did not parallel the accumulation and subsequent reduction in primary follicles during the early fertile lifespan of the mouse. Our results agree with established dogma that the initial endowment of ovarian follicles is not supplemented by an appreciable number of stem cells; rather, it is sufficient to ensure the fertility needs of the adult mouse.


Functional characterization of synthetic leukotriene B and its stereochemical isomers.
Leukotriene B (LTB), a potent lipid chemotactic factor for neutrophils, is 5S,12R-dihydroxy-6,14-cis,8,10-trans-eicosatetraenoic acid (Fig 1), based upon direct comparison of natural LTB with synthetic 5S,12R-dihydroxy-6,8,10,14-eicosatetraenoic acid (5,12-di-HETE) stereoisomers in three biological assays. Of the six synthetic stereoisomers evaluated, only the 5S,12R,6,14-cis,8,10-trans compound had chemotactic potency for human neutrophils in vitro that was comparable to that of natural LTB, with a concentration of 3 X 10(9-9) M eliciting a one-half maximum response. In contrast, the racemic mixture of 5R,12R- and 5S,12S-6,10-trans,8,14-cis, the racemic mixture of 5S,12R- and 5R,12S-6,10-trans,8,14-cis, the 5S,12R-6,8-trans,10,14-cis, the 5S,12R-6,8,10-trans,14-cis, and the 5S,12S-6,8,10-trans,14-cis stereoisomers required concentrations of 3 X 10(-7) to 1 X 10(-6) M to elicit comparable responses. Only natural LTB and its synthetic counterpart elicited a local neutrophil infiltration when injected into the skin of the rhesus monkey at 10 ng and 100 ng per site. Natural and synthetic LTB at a concentration of 3 X 10(-8) M each provoked an EC25 contractile response of guinea pig pulmonary parenchymal strips in vitro, whereas the other four tested stereoisomers of 5,12-di-HETE were inactive at this concentration. Structure-function analyses suggest that the neutrophil chemotactic activity depends critically upon the C-1 to C-12 domain, including the stereochemistry of the 6-,8-,and 10-olefinic bonds and the presence of both hydroxyl groups.


Reading acuity in children: evaluation and reliability using MNREAD charts.
To study reading acuity in children and assess the reliability of its measurement using the Italian version of the MNREAD reading chart. One hundred sixteen children from grades 3 to 8 with normal near vision were tested at a pediatric eye care institution in northeastern Italy. Visual acuity was evaluated with Early Treatment Diabetic Retinopathy Study (ETDRS) charts and reading acuity with MNREAD charts. Examination with charts 1 and 2 by the same examiner in the same visit were used as test-retest measurements. Mean improvement of performance from grades 3 to 8 was approximately 1 line for ETDRS acuity and 1.5 sentences for reading acuity and critical print size. Mean maximum reading speed increased from 96 to 167 words/minute. Regression analysis showed that a stable reading speed was obtained throughout sentences of decreasing size up to the critical print size. Measurement error, expressed as 95% limits of agreement, was +/- 0.136 logarithm of the minimum angle of resolution (logMAR) and +/- 0.142 logMAR for ETDRS acuity and reading acuity, respectively. Critical print size had the lowest reliability (+/- 0.193 logMAR). Maximum reading speed limits of agreement were +/- 0.077 logarithm of words per minute (logWPM), or a 19% change in words per minute. There were no detectable effects of grade and refractive error on measurement errors. In children, MNREAD charts provide measurements of reading acuity as reliable as ETDRS chart measurements of distance acuity on a logMAR scale. They also provide highly reliable measurements of the maximum reading speed across all grades, independent of its noticeable increase.


Marijuana smoking among school-aged adolescents in the Brčko District of Bosnia and Herzegovina: A cross-sectional study.
The aim was to determine the prevalence of marijuana smoking among school-aged adolescents in the Brčko District of Bosnia and Herzegovina, with particular regard to their gender, age and residence, and the frequency of marijuana smoking in the past thirty days in relation to their peers in the rest of Bosnia and Herzegovina, the Republic of Croatia and the Republic of Serbia. This research, designed as a cross-sectional study and based on the ESPAD (European School Survey Project on Alcohol and Other Drugs) questionnaire, adjusted to this research, encompassed 4,188 adolescents from elementary and secondary schools. The data were collected by means of questionnaires tailored to each respondent. A significantly lower number of adolescents smoke marijuana in comparison to those who do not smoke, but male adolescents smoke more often than female adolescents (p<0.001), as well as urban youth in comparison to rural youth (p=0.04). Every fourth adolescent, regardless of gender, who smoked marijuana, used it before the age of thirteen (p<0.001), male adolescents more often than females (p=0.002). In the previous thirty days a higher percentage of all the respondents from the Brčko District had smoked marijuana than those from the Republika Srpska and the RS (p<0.001), and there is no difference between them and their peers from the Federation of Bosnia and Herzegovina and the RC (p=0.382 and p=0.608). Smoking marijuana in the Brčko District is a major public health problem. Male adolescents smoke marijuana more often than female adolescents, and urban youth more in comparison to rural youth. In the previous thirty days adolescents from the Brčko District smoked more often than their peers from the Republic of Serbia and the Republika Srpska, and with the same intensity but less frequently compared to adolescents from the Republic of Croatia and the Federation of Bosnia and Herzegovina.


Neural correlates of relational memory: successful encoding and retrieval of semantic and perceptual associations.
Using event-related functional magnetic resonance imaging, we identified brain regions involved in successful relational memory (RM) during encoding and retrieval for semantic and perceptual associations or in general, independent of phase and content. Participants were scanned while encoding and later retrieving associations between pairs of words (semantic RM) or associations between words and fonts (perceptual RM). Encoding success activity (ESA) was identified by comparing study-phase activity for items subsequently remembered (hits) versus forgotten (misses) and retrieval success activity (RSA) by comparing test-phase activity for hits versus misses. The study yielded three main sets of findings. First, ESA-RSA differences were found within the medial temporal lobes (MTLs) and within the prefrontal cortex (PFC). Within the left MTL, ESA was greater in the anterior hippocampus, and RSA was greater in the posterior parahippocampal cortex/hippocampus. This finding is consistent with the notion of an encoding-retrieval gradient along the longitudinal MTL axis. Within the left PFC, ESA was greater in ventrolateral PFC, and RSA was greater in dorsolateral and anterior PFC. This is the first evidence of a dissociation in successful encoding and retrieval activity within left PFC. Second, consistent with the transfer-appropriate processing principle, some ESA regions were reactivated during RSA in a content-specific manner. For semantic RM, these regions included the left ventrolateral PFC, whereas for perceptual RM, they included occipitoparietal and right parahippocampal regions. Finally, only one region in the entire brain was associated with RM in general (i.e., for both semantic and perceptual ESA and RSA): the left hippocampus. This finding highlights the fundamental role of the hippocampus in RM.


The Evidence in Support of Physicians and Health Care Providers as Physical Activity Role Models.
Physical inactivity constitutes the fourth leading cause of death worldwide. Health care providers (HCPs) should play a key role in counseling and appropriately referring their patients to adopt physical activity (PA). Previous reports suggest that active HCPs are more likely to provide better, more credible, and motivating preventive counseling to their patients. This review summarizes the available evidence on the association between HCPs' personal PA habits and their related PA counseling practices. Based on relevant studies, a snowball search strategy identified, out of 196 studies screened, a total of 47 pertinent articles published between 1979 and 2012. Of those, 23 described HCPs' PA habits and/or their counseling practices and 24 analytic studies evaluated the association between HCPs' personal PA habits and their PA counseling practices. The majority of studies came from the United States (n = 33), and 9 studies included nonphysicians (nurses, pharmacists, and other HCPs). PA levels were mostly self-reported, and counseling was typically assessed as self-reported frequency or perceived self-efficacy in clinical practice. Most (19 out of 24) analytic studies reported a significant positive association between HCPs' PA habits and counseling frequency, with odds ratios ranging between 1.4 and 5.7 (P < .05), in 6 studies allowing direct comparison. This review found consistent evidence supporting the notion that physically active physicians and other HCPs are more likely to provide PA counseling to their patients and can indeed become powerful PA role models. This evidence appears sufficient to justify randomized trials to determine if adding interventions to promote PA among HCPs, also results in improvements in the frequency and quality of PA preventive counseling and referrals, delivered by HCPs, to patients in primary care settings. Future studies should also aim at objectively quantifying the effect of HCPs' PA role-modeling and how it influences patients' PA levels. More evidence from low-to-middle income countries is needed, where 80% of the deaths due to inactivity and related noncommunicable diseases already occur.


Detection of pathogenic elephant endotheliotropic herpesvirus in routine trunk washes from healthy adult Asian elephants (Elephas maximus) by use of a real-time quantitative polymerase chain reaction assay.
To investigate the pathogenesis and transmission of elephant endotheliotropic herpesvirus (EEHV1) by analyzing various elephant fluid samples with a novel EEHV1-specific real-time PCR assay. 5 apparently healthy captive Asian elephants (Elephas maximus) from the same herd. A real-time PCR assay was developed that specifically detects EEHV1. The assay was used to evaluate paired whole blood and trunk-wash samples obtained from the 5 elephants during a 15-week period. Deoxyribonucleic acid sequencing and viral gene subtyping analysis were performed on trunk-wash DNA preparations that had positive results for EEHV1. Viral gene subtypes were compared with those associated with past fatal cases of herpesvirus-associated disease within the herd. The PCR assay detected viral DNA to a level of 1,200 copies/mL of whole blood. It was used to detect EEHV1 in trunk secretions of 3 of the 5 elephants surveyed during the 15-week period. Viral gene subtyping analysis identified 2 distinct elephant herpesviruses, 1 of which was identical to the virus associated with a previous fatal case of herpesvirus-associated disease within the herd. EEHV1 was shed in the trunk secretions of healthy Asian elephants. Trunk secretions may provide a mode of transmission for this virus. Results of this study may be useful for the diagnosis, treatment, and management of EEHV1-associated disease and the overall management of captive elephant populations.


Swallowing function after stroke: prognosis and prognostic factors at 6 months.
Swallowing dysfunction (dysphagia) is common and disabling after acute stroke, but its impact on long-term prognosis for potential complications and the recovery from swallowing dysfunction remain uncertain. We aimed to prospectively study the prognosis of swallowing function over the first 6 months after acute stroke and to identify the important independent clinical and videofluoroscopic prognostic factors at baseline that are associated with an increased risk of swallowing dysfunction and complications. We prospectively assembled an inception cohort of 128 hospital-referred patients with acute first stroke. We assessed swallowing function clinically and videofluoroscopically, within a median of 3 and 10 days, respectively, of stroke onset, using standardized methods and diagnostic criteria. All patients were followed up prospectively for 6 months for the occurrence of death, recurrent stroke, chest infection, recovery of swallowing function, and return to normal diet. At presentation, a swallowing abnormality was detected clinically in 65 patients (51%; 95% CI, 42% to 60%) and videofluoroscopically in 82 patients (64%; 95% CI, 55% to 72%). During the subsequent 6 months, 26 patients (20%; 95% CI, 14% to 28%) suffered a chest infection. At 6 months after stroke, 97 of the 112 survivors (87%; 95% CI, 79% to 92%) had returned to their prestroke diet. Clinical evidence of a swallowing abnormality was present in 56 patients (50%; 95% CI, 40% to 60%). Videofluoroscopy was performed at 6 months in 67 patients who had a swallowing abnormality at baseline; it showed penetration of the false cords in 34 patients and aspiration in another 17. The single independent baseline predictor of chest infection during the 6-month follow-up period was a delayed or absent swallowing reflex (detected by videofluoroscopy). The single independent predictor of failure to return to normal diet was delayed oral transit (detected by videofluoroscopy). Independent predictors of the combined outcome event of swallowing impairment, chest infection, or aspiration at 6 months were videofluoroscopic evidence of delayed oral transit and penetration of contrast into the laryngeal vestibule, age >70 years, and male sex. Swallowing function should be assessed in all acute stroke patients because swallowing dysfunction is common, it persists in many patients, and complications frequently arise. The assessment of swallowing function should be both clinical and videofluoroscopic. The clinical and videofluoroscopic features at presentation that are important predictors of subsequent swallowing abnormalities and complications are videofluoroscopic evidence of delayed oral transit, a delayed or absent swallow reflex, and penetration. These findings require validation in other studies.


Patients' attitudes toward side effects of antidepressants: an Internet survey.
Patients' attitudes toward side effects of antidepressants are likely to differ according to gender, which has not yet been fully addressed in the literature. From the 228,310 registrants, 1,305 participants who had received antidepressant drugs within the past year were identified with the Yahoo Japan research monitor through four-step screening procedures. Participants were asked as to which side effect(s) they had experienced, whether they had reported those side effects to their physicians, and whether they had taken any action to counteract them. The questionnaire was completed by 1,187 participants. Side effects were reported in 73.4% of the participants; the prevalence of self-reported side effects was significantly higher in men than women (80.4% vs. 68.3%, P <0.05). The percentage of participants who reported side effects to their physicians widely differed depending on the nature of their experience, ranging from 45.7% to 89.9%; the lowest was for sexual dysfunction. The percentage of participants who had taken any action to relieve side effects varied among side effects from 26.3% for sexual dysfunction to 89.5% for dry mouth. Moreover, a lower percentage of women had reported sexual dysfunction to physicians (36.6% vs. 60.7%, P <0.05) and had taken any action to counteract the problem (19.8% vs. 36.9%, P <0.05). Given that patients experienced with antidepressants are likely to be reluctant to report sexual side effects, physicians should be cognizant of the potential presence of sexual dysfunction in patients who are taking antidepressants, especially for women.


Effect of plasmid DNA isoforms on preparative anion exchange chromatography.
Increased need for plasmid DNA (pDNA) with sizes above 10 kbp (large pDNA) in gene therapy and vaccination brings the need for its large-scale production with high purity. Chromatographic purification of large pDNA is often challenging due to low process yields and column clogging, especially using anion-exchanging columns. The goal of our investigation was to evaluate the mass balance and pDNA isoform composition at column outlet for plasmids of different sizes in combination with weak anion exchange (AEX) monolith columns of varying channel size (2, 3 and 6 µm channel size). We have proven that open circular pDNA (OC pDNA) isoform is an important driver of reduced chromatographic performance in AEX chromatography. The main reason for the behaviour is the entrapment of OC pDNA in chromatographic supports with smaller channel sizes. Entrapment of individual isoforms was characterised for porous beads and convective monolithic columns. Convective entrapment of OC pDNA isoform was confirmed on both types of stationary phases. Porous beads in addition showed a reduced recovery of supercoiled pDNA (on an 11.6 kbp plasmid) caused by diffusional entrapment within the porous structure. Use of convective AEX monoliths or membranes with channel diameter >3.5 µm has been shown to increase yields and prevent irreversible pressure build-up and column clogging during purification of plasmids at least up to 16 kbp in size.


Concussion in adolescence and risk of multiple sclerosis.
To assess whether concussion in childhood or adolescence is associated with subsequent multiple sclerosis (MS) risk. Previous research suggests an association, but methodological limitations included retrospective data collection and small study populations. The national Swedish Patient Register (hospital diagnoses) and MS Register were used to identify all MS diagnoses up to 2012 among people born since 1964, when the Patient Register was established. The 7,292 patients with MS were matched individually with 10 people without MS by sex, year of birth, age/vital status at MS diagnosis, and region of residence (county), resulting in a study population of 80,212. Diagnoses of concussion and control diagnoses of broken limb bones were identified using the Patient Register from birth to age 10 years or from age 11 to 20 years. Conditional logistic regression was used to examine associations with MS. Concussion in adolescence was associated with a raised risk of MS, producing adjusted odds ratios (95% confidence intervals) of 1.22 (1.05-1.42, p = 0.008) and 2.33 (1.35-4.04, p = 0.002) for 1 diagnosis of concussion and >1 diagnosis of concussion, respectively, compared with none. No notable association with MS was observed for concussion in childhood, or broken limb bones in childhood and adolescence. Head trauma in adolescence, particularly if repeated, is associated with a raised risk of future MS, possibly due to initiation of an autoimmune process in the central nervous system. This further emphasizes the importance of protecting young people from head injuries. Ann Neurol 2017;82:554-561.


Exome capture from saliva produces high quality genomic and metagenomic data.
Targeted capture of genomic regions reduces sequencing cost while generating higher coverage by allowing biomedical researchers to focus on specific loci of interest, such as exons. Targeted capture also has the potential to facilitate the generation of genomic data from DNA collected via saliva or buccal cells. DNA samples derived from these cell types tend to have a lower human DNA yield, may be degraded from age and/or have contamination from bacteria or other ambient oral microbiota. However, thousands of samples have been previously collected from these cell types, and saliva collection has the advantage that it is a non-invasive and appropriate for a wide variety of research. We demonstrate successful enrichment and sequencing of 15 South African KhoeSan exomes and 2 full genomes with samples initially derived from saliva. The expanded exome dataset enables us to characterize genetic diversity free from ascertainment bias for multiple KhoeSan populations, including new exome data from six HGDP Namibian San, revealing substantial population structure across the Kalahari Desert region. Additionally, we discover and independently verify thirty-one previously unknown KIR alleles using methods we developed to accurately map and call the highly polymorphic HLA and KIR loci from exome capture data. Finally, we show that exome capture of saliva-derived DNA yields sufficient non-human sequences to characterize oral microbial communities, including detection of bacteria linked to oral disease (e.g. Prevotella melaninogenica). For comparison, two samples were sequenced using standard full genome library preparation without exome capture and we found no systematic bias of metagenomic information between exome-captured and non-captured data. DNA from human saliva samples, collected and extracted using standard procedures, can be used to successfully sequence high quality human exomes, and metagenomic data can be derived from non-human reads. We find that individuals from the Kalahari carry a higher oral pathogenic microbial load than samples surveyed in the Human Microbiome Project. Additionally, rare variants present in the exomes suggest strong population structure across different KhoeSan populations.


Ensemble crowd perception: a viewpoint-invariant mechanism to represent average crowd identity.
Individuals can rapidly and precisely judge the average of a set of similar items, including both low-level (Ariely, 2001) and high-level objects (Haberman & Whitney, 2007). However, to date, it is unclear whether ensemble perception is based on viewpoint-invariant object representations. Here, we tested this question by presenting participants with crowds of sequentially presented faces. The number of faces in each crowd and the viewpoint of each face varied from trial to trial. This design required participants to integrate information from multiple viewpoints into one ensemble percept. Participants reported the mean identity of crowds (e.g., family resemblance) using an adjustable, forward-oriented test face. Our results showed that participants accurately perceived the mean crowd identity even when required to incorporate information across multiple face orientations. Control experiments showed that the precision of ensemble coding was not solely dependent on the length of time participants viewed the crowd. Moreover, control analyses demonstrated that observers did not simply sample a subset of faces in the crowd but rather integrated many faces into their estimates of average crowd identity. These results demonstrate that ensemble perception can operate at the highest levels of object recognition after 3-D viewpoint-invariant faces are represented.


Determinants of cognitive performance and decline in 20 diverse ethno-regional groups: A COSMIC collaboration cohort study.
With no effective treatments for cognitive decline or dementia, improving the evidence base for modifiable risk factors is a research priority. This study investigated associations between risk factors and late-life cognitive decline on a global scale, including comparisons between ethno-regional groups. We harmonized longitudinal data from 20 population-based cohorts from 15 countries over 5 continents, including 48,522 individuals (58.4% women) aged 54-105 (mean = 72.7) years and without dementia at baseline. Studies had 2-15 years of follow-up. The risk factors investigated were age, sex, education, alcohol consumption, anxiety, apolipoprotein E ε4 allele (APOE*4) status, atrial fibrillation, blood pressure and pulse pressure, body mass index, cardiovascular disease, depression, diabetes, self-rated health, high cholesterol, hypertension, peripheral vascular disease, physical activity, smoking, and history of stroke. Associations with risk factors were determined for a global cognitive composite outcome (memory, language, processing speed, and executive functioning tests) and Mini-Mental State Examination score. Individual participant data meta-analyses of multivariable linear mixed model results pooled across cohorts revealed that for at least 1 cognitive outcome, age (B = -0.1, SE = 0.01), APOE*4 carriage (B = -0.31, SE = 0.11), depression (B = -0.11, SE = 0.06), diabetes (B = -0.23, SE = 0.10), current smoking (B = -0.20, SE = 0.08), and history of stroke (B = -0.22, SE = 0.09) were independently associated with poorer cognitive performance (p < 0.05 for all), and higher levels of education (B = 0.12, SE = 0.02) and vigorous physical activity (B = 0.17, SE = 0.06) were associated with better performance (p < 0.01 for both). Age (B = -0.07, SE = 0.01), APOE*4 carriage (B = -0.41, SE = 0.18), and diabetes (B = -0.18, SE = 0.10) were independently associated with faster cognitive decline (p < 0.05 for all). Different effects between Asian people and white people included stronger associations for Asian people between ever smoking and poorer cognition (group by risk factor interaction: B = -0.24, SE = 0.12), and between diabetes and cognitive decline (B = -0.66, SE = 0.27; p < 0.05 for both). Limitations of our study include a loss or distortion of risk factor data with harmonization, and not investigating factors at midlife. These results suggest that education, smoking, physical activity, diabetes, and stroke are all modifiable factors associated with cognitive decline. If these factors are determined to be causal, controlling them could minimize worldwide levels of cognitive decline. However, any global prevention strategy may need to consider ethno-regional differences.


Acute physical exercise affected processing efficiency in an auditory attention task more than processing effectiveness.
Research on effects of acute physical exercise on performance in a concurrent cognitive task has generated equivocal evidence. Processing efficiency theory predicts that concurrent physical exercise can increase resource requirements for sustaining cognitive performance even when the level of performance is unaffected. This hypothesis was tested in a dual-task experiment. Sixty young adults worked on a primary auditory attention task and a secondary interval production task while cycling on a bicycle ergometer. Physical load (cycling) and cognitive load of the primary task were manipulated. Neither physical nor cognitive load affected primary task performance, but both factors interacted on secondary task performance. Sustaining primary task performance under increased physical and/or cognitive load increased resource consumption as indicated by decreased secondary task performance. Results demonstrated that physical exercise effects on cognition might be underestimated when only single task performance is the focus.


Abfraction: separating fact from fiction.
Non-carious cervical lesions involve loss of hard tissue and, in some instances, restorative material at the cervical third of the crown and subjacent root surface, through processes unrelated to caries. These non-carious processes may include abrasion, corrosion and possibly abfraction, acting alone or in combination. Abfraction is thought to take place when excessive cyclic, non-axial tooth loading leads to cusp flexure and stress concentration in the vulnerable cervical region of teeth. Such stress is then believed to directly or indirectly contribute to the loss of cervical tooth substance. This article critically reviews the literature for and against the concept of abfraction. Although there is theoretical evidence in support of abfraction, predominantly from finite element analysis studies, caution is advised when interpreting results of these studies because of their limitations. In fact, there is only a small amount of experimental evidence for abfraction. Clinical studies have shown associations between abfraction lesions, bruxism and occlusal factors, such as premature contacts and wear facets, but these investigations do not confirm causal relationships. Importantly, abfraction lesions have not been reported in pre-contemporary populations. It is important that oral health professionals understand that abfraction is still a theoretical concept, as it is not backed up by appropriate clinical evidence. It is recommended that destructive, irreversible treatments aimed at treating so-called abfraction lesions, such as occlusal adjustment, be avoided.


PTEN nuclear localization is regulated by oxidative stress and mediates p53-dependent tumor suppression.
The tumor suppressor gene PTEN (phosphatase and tensin homologue deleted on chromosome 10) is frequently mutated or deleted in various human cancers. PTEN localizes predominantly to the cytoplasm and functions as a lipid phosphatase, thereby negatively regulating the phosphatidylinositol 3-kinase-AKT signaling pathway. PTEN can also localize to the nucleus, where it binds and regulates p53 protein level and transcription activity. However, the precise function of nuclear PTEN and the factors that control PTEN nuclear localization are still largely unknown. In this study, we identified oxidative stress as one of the physiological stimuli that regulate the accumulation of nuclear PTEN. Specifically, oxidative stress inhibits PTEN nuclear export, a process depending on phosphorylation of its amino acid residue Ser-380. Nuclear PTEN, independent of its phosphatase activity, leads to p53-mediated G(1) growth arrest, cell death, and reduction of reactive oxygen species production. Using xenografts propagated from human prostate cancer cell lines, we reveal that nuclear PTEN is sufficient to reduce tumor progression in vivo in a p53-dependent manner. The data outlined in this study suggest a unique role of nuclear PTEN to arrest and protect cells upon oxidative damage and to regulate tumorigenesis. Since tumor cells are constantly exposed to oxidative stress, our study elucidates the cooperative roles of nuclear PTEN with p53 in tumor suppression.


Structural and Biochemical Investigations of the [4Fe-4S] Cluster-Containing Fumarate Hydratase from Leishmania major.
Class I fumarate hydratases (FHs) are central metabolic enzymes that use a [4Fe-4S] cluster to catalyze the reversible conversion of fumarate to S-malate. The parasite Leishmania major, which is responsible for leishmaniasis, expresses two class I FH isoforms: mitochondrial LmFH-1 and cytosolic LmFH-2. In this study, we present kinetic characterizations of both LmFH isoforms, present 13 crystal structures of LmFH-2 variants, and employ site-directed mutagenesis to investigate the enzyme's mechanism. Our kinetic data confirm that both LmFH-1 and LmFH-2 are susceptible to oxygen-dependent inhibition, with data from crystallography and electron paramagnetic resonance spectroscopy showing that oxygen exposure converts an active [4Fe-4S] cluster to an inactive [3Fe-4S] cluster. Our anaerobically conducted kinetic studies reveal a preference for fumarate over S-malate. Our data further reveal that single alanine substitutions of T467, R421, R471, D135, and H334 decrease kcat values 9-16000-fold without substantially affecting Km values, suggesting that these residues function in catalytic roles. Crystal structures of LmFH-2 variants are consistent with this idea, showing similar bidentate binding to the unique iron of the [4Fe-4S] cluster for substrate S-malate as observed in wild type FH. We further present LmFH-2 structures with substrate fumarate and weak inhibitors succinate and malonate bound in the active site and the first structure of an LmFH that is substrate-free and inhibitor-free, the latter showing increased mobility in the C-terminal domain. Collectively, these data provide insight into the molecular basis for the reaction catalyzed by LmFHs, enzymes that are potential drug targets against leishmaniasis.


Evaluation of urine CCA assays for detection of Schistosoma mansoni infection in Western Kenya.
Although accurate assessment of the prevalence of Schistosoma mansoni is important for the design and evaluation of control programs, the most widely used tools for diagnosis are limited by suboptimal sensitivity, slow turn-around-time, or inability to distinguish current from former infections. Recently, two tests that detect circulating cathodic antigen (CCA) in urine of patients with schistosomiasis became commercially available. As part of a larger study on schistosomiasis prevalence in young children, we evaluated the performance and diagnostic accuracy of these tests--the carbon test strip designed for use in the laboratory and the cassette format test intended for field use. In comparison to 6 Kato-Katz exams, the carbon and cassette CCA tests had sensitivities of 88.4% and 94.2% and specificities of 70.9% and 59.4%, respectively. However, because of the known limitations of the Kato-Katz assay, we also utilized latent class analysis (LCA) incorporating the CCA, Kato-Katz, and schistosome-specific antibody results to determine their sensitivities and specificities. The laboratory-based CCA test had a sensitivity of 91.7% and a specificity of 89.4% by LCA while the cassette test had a sensitivity of 96.3% and a specificity of 74.7%. The intensity of the reaction in both urine CCA tests reflected stool egg burden and their performance was not affected by the presence of soil transmitted helminth infections. Our results suggest that urine-based assays for CCA may be valuable in screening for S. mansoni infections.


Neurological and neuropsychological effects of low and moderate prenatal alcohol exposure.
Several explanations for the diverse results in research on foetal alcohol spectrum disorders or alcohol-related neurodevelopmental disorder might be at hand: timing, amount and patterns of alcohol exposure, as well as complex epigenetic responses. The genetic background of the offspring and its interaction with other prenatal and post-natal environmental cues are likely also of importance. In the present report, key findings about the possible effects of low and moderate doses of maternal alcohol intake on the neuropsychological development of the offspring are reviewed and plausible mechanisms discussed. Special focus is put on the serotonergic system within developmental and gene-environment frameworks. The review also suggests guidelines for future studies and also summarizes some of to-be-answered questions of relevance to clinical practice. Contradictory findings and paucity of studies on the effects of exposure to low alcohol levels during foetal life for the offspring's neuropsychological development call for large prospective studies, as well as for studies including neuroimaging and multi-omics analyses to dissect the neurobiological underpinnings of alcohol exposure-related phenotypes and to identify biomarkers. Finally, it remains to be investigated whether any safe threshold of alcohol drinking during pregnancy can be identified.


Target sequences for hunchback in a control region conferring Ultrabithorax expression boundaries.
Boundaries of Ultrabithorax expression are mediated by long-range repression acting through the PBX or ABX control region. We show here that either of these control regions confers an early band of beta-galactosidase expression which is restricted along the anteroposterior axis of the blastoderm embryo. This band is succeeded by a stripe pattern with very similar anteroposterior limits. Dissection of the PBX control region demonstrates that the two patterns are conferred by distinct cis-regulatory sequences contained within separate PBX subfragments. We find several binding sites for hunchback protein within both PBX subfragments. Zygotic hunchback function is required to prevent ectopic PBX expression. Moreover, the PBX pattern is completely suppressed in embryos containing uniformly distributed maternal hunchback protein. Our results strongly suggest that hunchback protein directly binds to the PBX control region and acts as a repressor to specify the boundary positions of the PBX pattern.


Results of a multisite survey of U.S. psychiatry residents on education in professionalism and ethics.
The authors assess the perspectives of psychiatry residents about the goals of receiving education in professionalism and ethics, how topics should be taught, and on what ethical principles the curriculum should be based. A written survey was sent to psychiatry residents (N=249) at seven U.S. residency programs in Spring 2005. The survey was based on an instrument originally developed at the University of New Mexico, consisting of 149 questions in 10 content domains, with 6 questions regarding ethics experiences during training and 5 demographic questions. A total of 151 psychiatry residents (61%) returned usable responses to our survey. Residents reported receiving a moderate amount of ethics training during medical school (mean: 5.20; scale: 1: None to 9: Very Much) and some ethics training during residency (mean: 4.60). Residents endorsed moderate to moderately-strong agreement with all 11 goals of medical education in professionalism and ethics (means: 5.29 to 7.49; scale: 1: Strongly Disagree to 9: Strongly Agree). Respondents were more likely to endorse the value of clinically- and expert-oriented learning methods over web-based educational approaches. U.S. psychiatry residents endorse a range of goals for education in professionalism and ethics. At the same time, they prefer that these topics be taught in clinically relevant ways and through expert instruction. The value of web-based approaches warrants further investigation.


Mirror agnosia.
Normal people rarely confuse the mirror image of an object with a real object so long as they realize they are looking into a mirror. We report a new neurological sign, 'mirror agnosia', following right parietal lesions in which this ability is severely compromised. We studied four right hemisphere stroke patients who had left visual field 'neglect'. i.e. they were indifferent to objects in their left visual field even though they were not blind. We then placed a vertical parasagittal mirror on each patients' right so that they could clearly see the reflection of objects placed in the (neglected) visual field. When shown a candy or pen on their left, the patients kept banging their hand into the mirror or groped behind it attempting to grab the reflection; they did not reach for the real object on the left, even though they were mentally quite lucid and knew they were looking into a mirror. Remarkably, all four patients kept complaining that the object was 'in the mirror', 'outside my reach' or 'behind the mirror'. Thus, even the patients' ability to make simple logical inferences about mirrors has been selectively warped to accommodate the strange new sensory world that they now inhabit. The finding may have implications for understanding how the brain creates representations of mirror reflections.


Computer-assisted reading intervention with a phonics approach for children using cochlear implants or hearing aids.
The present study examined computer-assisted reading intervention with a phonics approach for deaf and hard of hearing (DHH) children in Sweden using cochlear implants or hearing aids, or a combination of both. The study included 48 children, 5, 6 and 7 years of age. Sixteen children with normal hearing (NH) served as a reference group. The first purpose of the study was to compare NH and DHH children's reading ability at pre and post-intervention. The second purpose was to investigate effects of the intervention. Cognitive and demographic factors were analyzed in relation to reading improvement. Results showed no statistically significant difference for reading ability at the group level, although NH children showed overall higher reading scores at both test points. Age comparisons revealed a statistically significant higher reading ability in the NH 7-year-olds compared to the DHH 7-year-olds. The intervention proved successful for word decoding accuracy, passage comprehension and as a reduction of nonword decoding errors in both NH and DHH children. Reading improvement was associated with complex working memory and phonological processing skills in NH children. Correspondent associations were observed with visual working memory and letter knowledge in the DHH children. Age was the only demographic factor that was significantly correlated with reading improvement. The results suggest that DHH children's beginning reading may be influenced by visual strategies that might explain the reading delay in the older children.


Rhipicephalus appendiculatus (Acari: Ixodidae): dynamics of Thogoto virus infection in female ticks during feeding on guinea pigs.
Engorged nymphs (Rhipicephalus appendiculatus) were inoculated parenterally with Thogoto (THO) virus (approximately 1 microl per nymph; 10(6)-10(7) PFU/ml). The adult females which resulted were used as the source of infected ticks for this study. Hemolymph, salivary glands, synganglion, gut, ovary, and Malpighian tubules were collected on each day of the blood meal and titrated for THO virus by plaque assay. The percent of tissues infected with virus was 16% or less on the day of attachment. Percent infection rose for all tissues throughout 6-7 days of feeding, reaching 40-100% infection during the rapid phase of engorgement. For the first 4 days of feeding, virus titer in the synganglion was higher than in salivary glands (means of 6.4-34.7 PFU/synganglion and 1.6-8.8 PFU/salivary gland pair). From days 5-7, virus titer was generally higher in the salivary gland than the synganglion (means of 422, 408, and 817 PFU/gland pair and means of 62, 811, and 9 PFU/synganglion). However, because a salivary gland pair is much heavier than a synganglion, the virus concentration in the synganglion was much higher than in the salivary gland during the slow phase of feeding. During the rapid phase of feeding, the difference in virus titer between the synganglion and salivary gland reduced. This difference between the early and late stages of feeding may explain why a previous study [J. Gen. Virol. 70 (1989) 1093], using immunofluorescence and immuno-gold labelling, failed to detect virus in the salivary gland early in feeding. These data provide evidence to explain that R. appendiculatus can transmit THO virus within 24h of attachment, an important epidemiological finding.


Case report: infective endocarditis caused by Brevundimonas vesicularis.
There are few reports in the literature of invasive infection caused by Brevundimonas vesicularis in patients without immunosuppression or other predisposing factors. The choice of antimicrobial therapy for bacteremia caused by the pathogen requires more case experience to be determined. The case of a 40-year-old previously healthy man with subacute endocarditis proposed to be contributed from an occult dental abscess is described. The infection was found to be caused by B. vesicularis on blood culture results. The patient recovered without sequelae after treatment with ceftriaxone followed by subsequent ciprofloxacin therapy owing to an allergic reaction to ceftriaxone and treatment failure with ampicillin/sulbactam. To our knowledge, this is the first report of B. vesicularis as a cause of infective endocarditis. According to an overview of the literature and our experience, we suggest that third-generation cephalosporins, piperacillin/tazobactam, and ciprofloxacin are effective in treating invasive B. vesicularis infections, while the efficacy of ampicillin-sulbactam needs further evaluation.


Water-dispersed lamellar phases of symmetric poly(styrene)-block-poly(acrylic acid) diblock copolymers: model systems for flat dense polyelectrolyte brushes.
We investigate the static properties of a water-dispersed lamellar ( L) phase formed in the melt state with a nearly symmetric poly(styrene)-block-poly(acrylic acid) (PS- b-PAA) diblock copolymer. The PAA brush is considered as a model flat polyelectrolyte ( PE) brush of controlled surface density. Thanks to small-angle X-ray scattering, its behavior in water is studied as a function of (i) its ionization, through the pH of the dispersions which is increased by an addition of a known amount of a base, i.e. sodium hydroxyde NaOH, and (ii) in the presence of a monovalent salt, i.e. sodium chloride NaCl, of concentration C(S). At low pH, we find that the brush effectively behaves as a neutral brush. At high pH, the brush is in the so-called "osmotic regime", in which all sodium counterions are trapped within the brush volume and stretch the chains via an osmotic effect. The properties of such a brush in the presence of a monovalent salt, confirm this result, showing a C(S)(-1/3) dependence in the brush height L(O), in agreement with mean-field predictions. The L(O)- C(S) profiles at different ionizations give access to the actual brush internal charge fraction f. The results are found to be in very good quantitative agreement with experimental measures found in the literature, and can be completely and quantitatively described by Oosawa's approach to counterion condensation in a semi-dilute to concentrated solution of charged, rod-like chains.


Mindfulness-based cognitive therapy for sexuality (MBCT-S) improves sexual functioning and intimacy among older women with epilepsy: A multicenter randomized controlled trial.
Women with epilepsy (WWE) frequently experience sexual problems, including sexual dysfunction and sexual distress. Therefore, this study aimed to examine the efficacy of a mindfulness-based cognitive therapy for sexuality (MBCT-S) on sexual function and intimacy among older adult WWE in Iran. A multicenter randomized controlled trial was conducted at 15 neurology clinics in three Iranian cities. WWE together with their partners and healthcare providers were randomly assigned in patient and partner (PP; n = 220), patient, partner, and healthcare provider (PPHP; n = 220), or treatment as usual (TAU; n = 220) group. WWE and their partners in both the PP and PPHP groups received eight weeks of MBCT-S sessions. In the PPHP group, three individual sessions on sexual counseling were added for healthcare providers. Self-assessment scales were used at baseline, one month and six months after completing the intervention. Mixed linear regression models were developed to determine differences among the three groups. Multilevel mediation analyses were conducted to understand the mediating effects of mindfulness, intimacy, patient-physicians relationship and sexual counseling barriers. Improvements in sexual function, sexual distress, and intimacy were found in both the PP and PPHP groups at one-month and six-month follow-ups. The PPHP group compared with the PP group had greater improvement in intimacy at both follow-ups. Regarding sexual function, the PPHP group performed better than the PP group at the six-month follow-up. The MBCT-S appears efficacious in enhancing sexual function and reducing sex-related distress in Iranian older adult WWE. Future studies should examine its efficacy in other populations.


c-erbB-2 protein overexpression in breast cancer is a risk factor in patients with involved and uninvolved lymph nodes.
The c-erbB-2 gene is overexpressed in about 20% of human breast cancers. Four hundred and eighty-three cases previously examined by immunohistochemical staining for c-erbB-2 expression were analysed to assess the risk associated with the elevated protein expression. Oncoprotein expression was correlated with increasing tumour grade but not with oestrogen receptor status, nodal involvement, tumour size or age. There was an increased risk of relapse and death associated with c-erbB-2 expression irrespective of nodal involvement. This marker thus appears to be a significant prognostic factor in the early as well as the late stages of breast cancer.


Do piperacillin/tazobactam and other antibiotics with inhibitory activity against Clostridium difficile reduce the risk for acquisition of C. difficile colonization?
Systemic antibiotics vary widely in in vitro activity against Clostridium difficile. Some agents with activity against C. difficile (e.g., piperacillin/tazobactam) inhibit establishment of colonization in mice. We tested the hypothesis that piperacillin/tazobactam and other agents with activity against C. difficile achieve sufficient concentrations in the intestinal tract to inhibit colonization in patients. Point-prevalence culture surveys were conducted to compare the frequency of asymptomatic rectal carriage of toxigenic C. difficile among patients receiving piperacillin/tazobactam or other inhibitory antibiotics (e.g. ampicillin, linezolid, carbapenems) versus antibiotics lacking activity against C. difficile (e.g., cephalosporins, ciprofloxacin). For a subset of patients, in vitro inhibition of C. difficile (defined as a reduction in concentration after inoculation of vegetative C. difficile into fresh stool suspensions) was compared among antibiotic treatment groups. Of 250 patients, 32 (13 %) were asymptomatic carriers of C. difficile. In comparison to patients receiving non-inhibitory antibiotics or prior antibiotics within 90 days, patients currently receiving piperacillin/tazobactam were less likely to be asymptomatic carriers (1/36, 3 versus 7/36, 19 and 15/69, 22 %, respectively; P = 0.024) and more likely to have fecal suspensions with in vitro inhibitory activity against C. difficile (20/28, 71 versus 3/11, 27 and 4/26, 15 %; P = 0.03). Patients receiving other inhibitory antibiotics were not less likely to be asymptomatic carriers than those receiving non-inhibitory antibiotics. Our findings suggest that piperacillin/tazobactam achieves sufficient concentrations in the intestinal tract to inhibit C. difficile colonization during therapy.


Bilateral ureteric obstruction secondary to renal papillary necrosis.
A 2-year-old male presented to hospital with a 5-day history of vomiting and pyrexia. He was initially treated with non-steroidal medication as an anti-pyretic. Initial investigations demonstrated a raised urea and creatinine and he was treated with intravenous fluids. Within 24 h he became anuric with progressive renal insufficiency. Ultrasound scan demonstrated minimal bilateral hydronephrosis with debris in the lower pole calyces. The bladder was empty. Cystoscopy and retrograde contrast imaging revealed bilateral ureteric obstruction. Double J stents were inserted and his renal function returned to normal within 4 days. We believe the aetiology to be renal papillary necrosis and bilateral ureteric obstruction secondary to the administration of ibuprofen in association with dehydration.


Prediction of the financial performance of Ontario hospitals: a test of environmental determinist and adaptationist perspectives.
While other industries for many years have been concerned with the problem of financial distress, it is only recently that this issue has become a matter of interest to hospital managers, policy makers, and the general public. However, the determinants of hospital financial performance are neither well studied nor understood. The objectives of this study were to identify factors that affect the financial performance of Ontario hospitals and to construct a model that could be used to predict financial performance in the future. A number of organization and environmental factors that could influence financial performance were postulated and then tested for their statistical impact and predictive ability. Cross-sectional data over the 3-year-period 1986-1988 for 223 Ontario public hospitals were used. The first 2 years of data served as a derivation sample for hypothesis testing and development of a predictive model. The third year of data was used as a holdout sample for cross validation. Information on the variables investigated came from secondary sources, in particular Statistics Canada's Annual Hospital Returns. Univariate analyses revealed distressed hospitals were more likely to earn more revenues from non-government sources, to be non-teaching institutions and have longer chronic lengths of stay, and to be found in areas with higher per capita incomes, number of females in the population, physician supply, and area wage rates. A five variable prediction model was developed which accounted for 25% of the variance in financial performance in the derivation sample and on cross validation dropped to 21%. The model identified greater hospital size, older plants, higher technological complexity, more intensive care services, and location in areas with more females to be significant predictors of financial distress. Overall, environmental factors (community and structural characteristics) were more important in influencing financial performance. The implication for hospital managers is to underscore that an important dimension of successful leadership requires they remain outwardly focused and involved in managing the external environment. For policy makers the need is to develop funding formulae which encourage efficiency and are also responsive to differences in community and structural characteristics across hospitals.


Development of a Searchable Database of Cryoablation Simulations for Use in Treatment Planning.
To create and validate a planning tool for multiple-probe cryoablation, using simulations of ice ball size and shape for various ablation probe configurations, ablation times, and types of tissue ablated. Ice ball size and shape was simulated using the Pennes bioheat equation. Five thousand six hundred and seventy different cryoablation procedures were simulated, using 1-6 cryoablation probes and 1-2 cm spacing between probes. The resulting ice ball was measured along three perpendicular axes and recorded in a database. Simulated ice ball sizes were compared to gel experiments (26 measurements) and clinical cryoablation cases (42 measurements). The clinical cryoablation measurements were obtained from a HIPAA-compliant retrospective review of kidney and liver cryoablation procedures between January 2015 and February 2016. Finally, we created a web-based cryoablation planning tool, which uses the cryoablation simulation database to look up the probe spacing and ablation time that produces the desired ice ball shape and dimensions. Average absolute error between the simulated and experimentally measured ice balls was 1 mm in gel experiments and 4 mm in clinical cryoablation cases. The simulations accurately predicted the degree of synergy in multiple-probe ablations. The cryoablation simulation database covers a wide range of ice ball sizes and shapes up to 9.8 cm. Cryoablation simulations accurately predict the ice ball size in multiple-probe ablations. The cryoablation database can be used to plan ablation procedures: given the desired ice ball size and shape, it will find the number and type of probes, probe configuration and spacing, and ablation time required.


Effectiveness of a Statewide Abusive Head Trauma Prevention Program in North Carolina.
Abusive head trauma (AHT) is a serious condition, with an incidence of approximately 30 cases per 100,000 person-years in the first year of life. To assess the effectiveness of a statewide universal AHT prevention program. In total, 88.29% of parents of newborns (n = 405 060) in North Carolina received the intervention (June 1, 2009, to September 30, 2012). A comparison of preintervention and postintervention was performed using nurse advice line telephone calls regarding infant crying (January 1, 2005, to December 31, 2010). A difference-in-difference analysis compared AHT rates in the prevention program state with those of other states before and after the implementation of the program (January 1, 2000, to December 31, 2011). The Period of PURPLE Crying intervention, developed by the National Center on Shaken Baby Syndrome, was delivered by nurse-provided education, a DVD, and a booklet, with reinforcement by primary care practices and a media campaign. Changes in proportions of telephone calls for crying concerns to a nurse advice line and in AHT rates per 100,000 infants after the intervention (June 1, 2009, to September 30, 2011) in the first year of life using hospital discharge data for January 1, 2000, to December 31, 2011. In the 2 years after implementation of the intervention, parental telephone calls to the nurse advice line for crying declined by 20% for children younger than 3 months (rate ratio, 0.80; 95% CI, 0.73-0.87; P < .001) and by 12% for children 3 to 12 months old (rate ratio, 0.88; 95% CI, 0.78-0.99; P = .03). No reduction in state-level AHT rates was observed, with mean rates of 34.01 person-years before the intervention and 36.04 person-years after the intervention. A difference-in-difference analysis from January 1, 2000, to December 31, 2011, controlling for economic indicators, indicated that the intervention did not have a statistically significant effect on AHT rates (β coefficient, -1.42; 95% CI, -13.31 to 10.45). The Period of PURPLE Crying intervention was associated with a reduction in telephone calls to a nurse advice line. The study found no reduction in AHT rates over time in North Carolina relative to other states. Consequently, while this observational study was feasible and supported the program effectiveness in part, further programmatic efforts and evaluation are needed to demonstrate an effect on AHT rates.


Bistability in one equation or fewer.
When several genes or proteins modulate one another's activity as part of a network, they sometimes produce behaviors that no protein could accomplish on its own. Intuition for these emergent behaviors often cannot be obtained simply by tracing causality through the network in discreet steps. Specifically, when a network contains a feedback loop, biologists need specialized tools to understand the network's behaviors and their necessary conditions. This analysis is grounded in the mathematics of ordinary differential equations. We, however, will demonstrate the use of purely graphical methods to determine, for experimental data, the plausibility of two network behaviors, bistability and irreversibility. We use the Xenopus laevis oocyte maturation network as our example, and we make special use of iterative stability analysis, a graphical tool for determining stability in two dimensions.


Dynamics of arbuscular mycorrhizal fungal community structure and functioning along a nitrogen enrichment gradient in an alpine meadow ecosystem.
Nitrogen (N) availability is increasing dramatically in many ecosystems, but the influence of elevated N on the functioning of arbuscular mycorrhizal (AM) fungi in natural ecosystems is not well understood. We measured AM fungal community structure and mycorrhizal function simultaneously across an experimental N addition gradient in an alpine meadow that is limited by N but not by phosphorus (P). AM fungal communities at both whole-plant-community (mixed roots) and single-plant-species (Elymus nutans roots) scales were described using pyro-sequencing, and the mycorrhizal functioning was quantified using a mycorrhizal-suppression treatment in the field (whole-plant-community scale) and a glasshouse inoculation experiment (single-plant-species scale). Nitrogen enrichment progressively reduced AM fungal abundance, changed AM fungal community composition, and shifted mycorrhizal functioning towards parasitism at both whole-plant-community and E. nutans scales. N-induced shifts in AM fungal community composition were tightly linked to soil N availability and/or plant species richness, whereas the shifts in mycorrhizal function were associated with the communities of specific AM fungal lineages. The observed changes in both AM fungal community structure and functioning across an N enrichment gradient highlight that N enrichment of ecosystems that are not P-limited can induce parasitic mycorrhizal functioning and influence plant community structure and ecosystem sustainability.