Soluble urokinase plasminogen activator receptor (suPAR) levels in healthy pregnancy and preeclampsia.
Preeclampsia is characterized by a maternal systemic inflammatory response and the impairment of maternal immune tolerance present in healthy pregnancy. Soluble urokinase plasminogen activator receptor (suPAR) is a biomarker increasingly used for the monitoring of systemic inflammation. We aimed to assess the levels of suPAR and other markers of systemic inflammation in preeclampsia compared to healthy pregnancy. We determined plasma suPAR, IL-6 and high sensitivity C-reactive protein (hs-CRP) levels in plasma samples of 62 healthy pregnant and 41 preeclamptic women in the third trimester of pregnancy. Plasma suPAR levels were elevated in preeclampsia [3.18 (2.30-4.71) ng/mL vs. 2.02 (1.81-2.40) ng/mL, p=0.0001, median (interquartile range)]. IL-6 and hs-CRP levels were also higher compared with healthy pregnancy [5.99 (2.97-18.12) pg/mL vs. 1.41 (1.00-2.70) pg/mL, p=0.0001 and 6.60 (3.55-15.40) mg/L vs. 3.90 (2.10-7.25) mg/L, p=0.006, respectively, median (interquartile range)]. Linear regression analyses revealed an association between individual plasma suPAR and log IL-6 levels as well as log hs-CRP levels. suPAR levels are elevated in preeclampsia and vary in a narrower range compared with IL-6 and hs-CRP. ROC analysis indicated that monitoring of suPAR levels is a suitable tool for the detection of systemic inflammation in pregnancy.


Vascular responses to ouabain and norepinephrine in low and normal renin hypertension.
A circulating Na+, K+-ATPase inhibitor may cause arterial hypertension in patients with suppressed plasma renin activity, either directly or by sensitizing peripheral vessels to alpha-adrenergic stimulation. This hypothesis was tested by evaluating forearm arteriolar (plethysmographic technique) response to exogenous alpha-adrenergic stimulation by a 2-minute intra-arterial infusion of norepinephrine (0.1 microgram/dl tissue per minute) and to Na+, K+-ATPase inhibition by sequential 20-minute intra-arterial infusions of ouabain (0.36 and 0.72 microgram/dl tissue per minute). Two groups of hypertensive subjects with suppressed plasma renin activity, either essential or secondary to aldosterone excess, were compared with age-matched and sex-matched hypertensive subjects with normal plasma renin activity (n = 7 per group). No significant differences in forearm vascular response to norepinephrine were found among the three groups. Ouabain caused a highly significant, dose-related increment in forearm vascular resistance that was not accompanied by changes in the contralateral limb or systemic blood pressure. No significant interindividual differences in vascular responsiveness to ouabain were found. The individual increments in forearm vascular resistance during ouabain administration were unrelated to basal values or to plasma aldosterone, norepinephrine, or potassium concentrations. These data are not consistent with the hypothesis that suppressed basal Na+, K+-ATPase activity is primarily a characteristic of hypertensive patients with unresponsive plasma renin activity. Overall, these results cast doubts on the possibility of linking the development of human low renin hypertension to an endogenous Na+, K+-ATPase inhibitor.


Blocking Synthesis of the Variant Surface Glycoprotein Coat in Trypanosoma brucei Leads to an Increase in Macrophage Phagocytosis Due to Reduced Clearance of Surface Coat Antibodies.
The extracellular bloodstream form parasite Trypanosoma brucei is supremely adapted to escape the host innate and adaptive immune system. Evasion is mediated through an antigenically variable Variant Surface Glycoprotein (VSG) coat, which is recycled at extraordinarily high rates. Blocking VSG synthesis triggers a precytokinesis arrest where stalled cells persist for days in vitro with superficially intact VSG coats, but are rapidly cleared within hours in mice. We therefore investigated the role of VSG synthesis in trypanosome phagocytosis by activated mouse macrophages. T. brucei normally effectively evades macrophages, and induction of VSG RNAi resulted in little change in phagocytosis of the arrested cells. Halting VSG synthesis resulted in stalled cells which swam directionally rather than tumbling, with a significant increase in swim velocity. This is possibly a consequence of increased rigidity of the cells due to a restricted surface coat in the absence of VSG synthesis. However if VSG RNAi was induced in the presence of anti-VSG221 antibodies, phagocytosis increased significantly. Blocking VSG synthesis resulted in reduced clearance of anti-VSG antibodies from the trypanosome surface, possibly as a consequence of the changed motility. This was particularly marked in cells in the G2/ M cell cycle stage, where the half-life of anti-VSG antibody increased from 39.3 ± 4.2 seconds to 99.2 ± 15.9 seconds after induction of VSG RNAi. The rates of internalisation of bulk surface VSG, or endocytic markers like transferrin, tomato lectin or dextran were not significantly affected by the VSG synthesis block. Efficient elimination of anti-VSG-antibody complexes from the trypanosome cell surface is therefore essential for trypanosome evasion of macrophages. These experiments highlight the essentiality of high rates of VSG recycling for the rapid removal of host opsonins from the parasite surface, and identify this process as a key parasite virulence factor during a chronic infection.


Impact of prolapse meshes on the metabolism of vaginal extracellular matrix in rhesus macaque.
The impact of polypropylene mesh implantation on vaginal collagen and elastin metabolism was analyzed using a nonhuman primate model to further delineate the mechanism of mesh induced complications. Forty-nine middle-aged parous rhesus macaques underwent surgical implantation of 3 synthetic meshes via sacrocolpopexy. Gynemesh PS (n = 12) (Ethicon, Somerville, NJ) and 2 lower-weight, higher-porosity, lower-stiffness meshes (UltraPro [n = 19] [Ethicon] and Restorelle [n = 8] [Coloplast, Minneapolis, MN]) were implanted, in which UltraPro was implanted with its blue orientation lines perpendicular (low stiffness direction, n = 11) and parallel (high stiffness direction, n = 8) to the longitudinal axis of the vagina. Sham-operated animals were used as controls (n = 10). Twelve weeks after surgery, the mesh-tissue complex was excised and analyzed. Relative to sham, Gynemesh PS had a negative impact on the metabolism of both collagen and elastin-favoring catabolic reactions, whereas UltraPro induced an increase only in elastin degradation. Restorelle had the least impact. As compared with sham, the degradation of collagen and elastin in the vagina implanted with Gynemesh PS was increased with a simultaneous increase in active matrix metalloproteinase (MMP)-1, -8, -13, and total MMP-2 and -9 (all P < .05). The degradation of elastin (tropoelastin and mature elastin) was increased in the UltraPro-implanted vagina with a concomitant increase of MMP-2, and -9 (all P < .05). Collagen subtype ratio III/I was increased in Gynemesh PS and UltraPro perpendicular groups (P < .05). Following implantation with the heavier, less porous, and stiffer mesh, Gynemesh PS, the degradation of vaginal collagen and elastin exceeded synthesis, most likely as a result of increased activity of MMPs, resulting in a structurally compromised tissue.


Estimating the emigration rates of legal immigrants using administrative and survey data: the 1971 cohort of immigrants to the United States.
Based on administrative and survey data as well as data-based assumptions about the bounds on alien address reporting, this study provides estimates of the lower and upper bounds for the cumulative net emigration rates, by country and area of origin, of the FY1971 cohort of legal immigrants to the United States as of January 1979. The merged data indicate that the cumulative net emigration rate for the entire cohort could have been as high as 50 percent. Canadian emigration was probably between 51 and 55 percent. Emigration rates for legal immigrants from Central America, the Caribbean (excluding Cuba), and South America were at least as high as 50 percent, and could have been as high as 70 percent. Emigration rates for Koreans and Chinese could not have exceeded 22 percent over the same period.


Synthesis and Biological Screening of New Lawson Derivatives as Selective Substrate-Based Inhibitors of Cytochrome bo3 Ubiquinol Oxidase from Escherichia coli.
The respiratory chain of Escherichia coli contains two different types of terminal oxidase that are differentially regulated as a response to changing environmental conditions. These oxidoreductases catalyze the reduction of molecular oxygen to water and contribute to the proton motive force. The cytochrome bo3 oxidase (cyt bo3 ) acts as the primary terminal oxidase under atmospheric oxygen levels, whereas the bd-type oxidase is most abundant under microaerobic conditions. In E. coli, both types of respiratory terminal oxidase (HCO and bd-type) use ubiquinol-8 as electron donor. Here, we assess the inhibitory potential of newly designed and synthesized 3-alkylated Lawson derivatives through L-proline-catalyzed three-component reductive alkylation (TCRA). The inhibitory effects of these Lawson derivatives on the terminal oxidases of E. coli (cyt bo3 and cyt bd-I) were tested potentiometrically. Four compounds were able to reduce the oxidoreductase activity of cyt bo3 by more than 50 % without affecting the cyt bd-I activity. Moreover, two inhibitors for both cyt bo3 and cyt bd-I oxidase could be identified. Based on molecular-docking simulations, we propose binding modes of the new Lawson inhibitors. The molecular fragment benzyl enhances the inhibitory potential and selectivity for cyt bo3 , whereas heterocycles reduce this effect. This work extends the library of 3-alkylated Lawson derivatives as selective inhibitors for respiratory oxidases and provides molecular probes for detailed investigations of the mechanisms of respiratory-chain enzymes of E. coli.


One-visit endodontic retreatment of combined external/internal root resorption using a calcium-enriched mixture.
Combined external and internal root resorption (ERR/IRR) is a rare endodontic disease that leads to alteration of the root canal anatomy. This report describes the management of an endodontically failed molar that was severely affected by ERR/IRR. Radiographic examination demonstrated inadequate obturation of the root canals associated with ERR/IRR and a large periradicular lesion. During nonsurgical endodontic retreatment, the root canals were subjected to conventional chemomechanical debridement. In the same session, the entire distal root canal was obturated using a calcium-enriched mixture (CEM) cement, and the mesial canals were obturated with gutta-percha/sealer. The clinical findings and follow-up radiographs indicated favorable treatment outcomes after 12 months. One-visit application of CEM cement could be a successful approach for the management of combined ERR/IRR.


Tonsil ultrasound: technical approach and spectrum of pediatric peritonsillar infections.
Peritonsillar infections are one of the most common deep neck space infections, particularly in adolescents. Inaccurate diagnosis can lead to delay in management and potentially life-threatening complications. Contrast-enhanced computed tomography (CT) scan of the neck traditionally has been used to diagnose suspected peritonsillar abscess. With growing concern over radiation exposure, there has been increasing utilization of ultrasound (US) using intraoral and transcutaneous approaches. We chose the transcutaneous US technique due to its ease of performance in children. The purpose of this article is twofold: a) to describe our technique of performing transcutaneous US of the tonsil showing sonographic appearance of normal tonsil, highlighting pertinent anatomy and unique considerations for this modality in children, and b) to illustrate the sonographic findings in the spectrum of pediatric peritonsillar infections, which includes uncomplicated tonsillitis, peritonsillar cellulitis, small intratonsillar abscess and frank peritonsillar abscess. Parapharyngeal abscess can sometimes be detected.


Computed Tomography Aortic Valve Calcium Scoring in Patients With Aortic Stenosis.
Computed tomography aortic valve calcium scoring (CT-AVC) holds promise for the assessment of patients with aortic stenosis (AS). We sought to establish the clinical utility of CT-AVC in an international multicenter cohort of patients. Patients with AS who underwent ECG-gated CT-AVC within 3 months of echocardiography were entered into an international, multicenter, observational registry. Optimal CT-AVC thresholds for diagnosing severe AS were determined in patients with concordant echocardiographic assessments, before being used to arbitrate disease severity in those with discordant measurements. In patients with long-term follow-up, we assessed whether CT-AVC thresholds predicted aortic valve replacement and death. In 918 patients from 8 centers (age, 77±10 years; 60% men; peak velocity, 3.88±0.90 m/s), 708 (77%) patients had concordant echocardiographic assessments, in whom CT-AVC provided excellent discrimination for severe AS (C statistic: women 0.92, men 0.89). Our optimal sex-specific CT-AVC thresholds (women 1377 Agatston unit and men 2062 Agatston unit) were nearly identical to those previously reported (women 1274 Agatston unit and men 2065 Agatston unit). Clinical outcomes were available in 215 patients (follow-up 1029 [126-2251] days). Sex-specific CT-AVC thresholds independently predicted aortic valve replacement and death (hazard ratio, 3.90 [95% confidence interval, 2.19-6.78]; P<0.001) after adjustment for age, sex, peak velocity, and aortic valve area. Among 210 (23%) patients with discordant echocardiographic assessments, there was considerable heterogeneity in CT-AVC scores, which again were an independent predictor of clinical outcomes (hazard ratio, 3.67 [95% confidence interval, 1.39-9.73]; P=0.010). Sex-specific CT-AVC thresholds accurately identify severe AS and provide powerful prognostic information. These findings support their integration into routine clinical practice. URL: http://www.clinicaltrials.gov. Unique identifiers: NCT01358513, NCT02132026, NCT00338676, NCT00647088, NCT01679431.


Adenovirus transcription. II. RNA sequences complementary to simian virus 40 and adenovirus 2DNA in AD2+ND1- and AD2+ND3-infected cells.
The genomes of the two nondefective adenovirus 2/simian virus 40 (Ad2/SV 40) hybrid viruses, nondefective Ad2/SV 40 hybrid virus 1 (Ad2+ND1) and nondefective hybrid virus 3 (Ad2+ND3), WERE FORMED BY A DELETION OF ABOUT 5% OF Ad2 DNA and insertion of part of the SV40 genome. We have compared the cytoplasmic RNA synthesized during both the early and late stages of lytic infection of human cells by these hybrid viruses to that expressed in Ad2-infected and SV40-infected cells. Separated strands of the six fragments of 32P-labeled Ad2 DNA produced by cleavage with the restriction endonuclease EcoRI (isolated from Escherichia coli) and the four fragments of 32P-labeled SV40 DNA produced by cleavage with both a restriction nuclease isolated from Haemophilus parainfluenzae, Hpa1, and EcoRI were prepared by electrophoresis of denatured DNA in agarose gels. The fraction of each fragment strand expressed as cytoplasmic RNA was determined by annealing fragmented 32P-labeled strands to an excess of cellular RNA extracted from infected cells. The segment of Ad2 DNA deleted from both hybrid virus genomes is transcribed into cytoplasmic mRNA during the early phase of Ad2 infection. Hence, we suggest that Ad2 codes for at least one "early" gene product which is nonessential for virus growth in cell culture. In both early Ad2+ND1 and Ad2+ND3-infected cells, 1,000 bases of Ad2 DNA adjacent to the integrated SV40 sequences are expressed as cytoplasmic RNA but are not similarly expressed in early Ad2-infected cells. The 3' termini of this early hybrid virus RNA maps in the vicinity of 0.18 on the conventional SV40 map and probably terminates at the same position as early lytic SV40 cytoplasmic RNA. Therefore, the base sequence in this region of SV40 DNA specifies the 3' termini of early messenger RNA present in both hybrid virus and SV40-infected cells.


Using Longitudinal Data to Understand Changes in Consistent Contraceptive Use.
Most studies of contraceptive behavior rely on cross-sectional data and are unable to adequately measure fluctuations in contraceptive use or changes in circumstances and attitudes that are likely to be associated with this outcome. Between November 2012 and May 2014, four waves of data were gathered from a national sample of 1,842 women aged 18-39 at baseline. Cross-tabulations were used to examine change and stability in time-varying characteristics theorized to be associated with consistent contraceptive use. Random-effects and fixed-effects logistic regression models were used to examine variables associated with consistent contraceptive use. While a majority of women were at risk of unintended pregnancy during each survey period, only 42% were at risk during all four. Random-effects logistic regression analysis revealed that the odds of being a consistent contraceptive user were 10 times as high for a woman who expressed a strong pregnancy avoidance attitude as for a woman who had a weak attitude. This strong association was confirmed in the fixed-effects model. However, having a strong desire to avoid pregnancy was not static; among women at risk of unintended pregnancy during at least one survey period, 53% reported a change in attitude. These findings build on prior research suggesting that pregnancy avoidance attitudes are an important motivator for contraceptive use. It is critical to recognize that the context in which many women make decisions about pregnancy and contraceptive use changes over relatively short periods of time.


L1 and HERV-W retrotransposons are hypomethylated in human ovarian carcinomas.
Wide-spread hypomethylation of CpG dinucleotides is characteristic of many cancers. Retrotransposons have been identified as potential targets of hypomethylation during cellular transformation. We report the results of an preliminary examination of the methylation status of CpG dinucleotides associated with the L1 and HERV-W retrotransposons in benign and malignant human ovarian tumors. We find a reduction in the methylation of CpG dinucleotides within the promoter regions of these retroelements in malignant relative to non-malignant ovarian tissues. Consistent with these results, we find that relative L1 and HERV-W expression levels are elevated in representative samples of malignant vs. non-malignant ovarian tissues.


Acute chest pain in the emergency room. Identification and examination of low-risk patients.
Clinical and laboratory data from 596 patients who came to an emergency room complaining of chest pain indicated that no single variable could identify low-risk patients as well as a normal ECG. A combination of three variables--sharp or stabbing pain, no history of angina or myocardial infarction, and pain with pleuritic or positional components or pain that was reproduced by palpation of the chest wall--defined a very-low-risk group in which ECGs did not add accuracy to the evaluation and were potentially misleading; however, only 48 patients (8%) fell into this category. Standard cardiac enzyme levels were of almost no use as an emergency room indicator of myocardial infarction. These findings emphasize the difficulty of identifying patients at low risk for myocardial infarction or unstable angina in the emergency room without consideration of many factors from the history, the physical examination, and the ECG.


RB loss contributes to aggressive tumor phenotypes in MYC-driven triple negative breast cancer.
Triple negative breast cancer (TNBC) is characterized by multiple genetic events occurring in concert to drive pathogenic features of the disease. Here we interrogated the coordinate impact of p53, RB, and MYC in a genetic model of TNBC, in parallel with the analysis of clinical specimens. Primary mouse mammary epithelial cells (mMEC) with defined genetic features were used to delineate the combined action of RB and/or p53 in the genesis of TNBC. In this context, the deletion of either RB or p53 alone and in combination increased the proliferation of mMEC; however, the cells did not have the capacity to invade in matrigel. Gene expression profiling revealed that loss of each tumor suppressor has effects related to proliferation, but RB loss in particular leads to alterations in gene expression associated with the epithelial-to-mesenchymal transition. The overexpression of MYC in combination with p53 loss or combined RB/p53 loss drove rapid cell growth. While the effects of MYC overexpression had a dominant impact on gene expression, loss of RB further enhanced the deregulation of a gene expression signature associated with invasion. Specific RB loss lead to enhanced invasion in boyden chambers assays and gave rise to tumors with minimal epithelial characteristics relative to RB-proficient models. Therapeutic screening revealed that RB-deficient cells were particularly resistant to agents targeting PI3K and MEK pathway. Consistent with the aggressive behavior of the preclinical models of MYC overexpression and RB loss, human TNBC tumors that express high levels of MYC and are devoid of RB have a particularly poor outcome. Together these results underscore the potency of tumor suppressor pathways in specifying the biology of breast cancer. Further, they demonstrate that MYC overexpression in concert with RB can promote a particularly aggressive form of TNBC.


Transforming growth factor alpha expression drives constitutive epidermal growth factor receptor pathway activation and sensitivity to gefitinib (Iressa) in human pancreatic cancer cell lines.
The epidermal growth factor receptor (EGFR) is considered an important therapeutic target in pancreatic cancer, but it is currently impossible to identify those patients who are most likely to benefit from EGFR-directed therapy. We examined the biological effects of the EGFR tyrosine kinase inhibitor gefitinib (ZD1839, Iressa) in a panel of nine human pancreatic cancer cell lines. The drug strongly inhibited DNA synthesis and induced low levels of apoptosis at clinically relevant concentrations in a subset of three of the lines (L3.6pl, BxPC3, and Cfpac1). Sensitivity to gefitinib correlated directly with ligand [transforming growth factor-alpha (TGF-alpha)] expression (r(2) = 0.71, P = 0.004) but not with surface EGFR expression. The gefitinib-sensitive cells displayed constitutive baseline EGFR phosphorylation, whereas the gefitinib-resistant cells did not. Exposure to gefitinib or a small interfering RNA construct specific for TGF-alpha reversed the constitutive EGFR phosphorylation and downstream target [extracellular signal-regulated kinases (ERK), AKT] phosphorylation in the gefitinib-sensitive cells but had no effects on ERK or AKT phosphorylation in gefitinib-resistant cells. Baseline EGFR phosphorylation was lower in a subclone of L3.6pl selected for low TGF-alpha expression, and these cells were also resistant to gefitinib-mediated growth inhibition. Gefitinib blocked the growth of tumor xenografts derived from L3.6pl cells but had no effect on the growth of tumors derived from EGFR-independent MiaPaCa-2 cells. Together, our data show that TGF-alpha expression identifies a subset of human pancreatic cancer cells that is dependent on EGFR signaling in vitro and in vivo. Quantification of TGF-alpha expression may therefore represent an effective means of identifying EGFR-responsive primary tumors.


Calcification rates and the effect of ocean acidification on Mediterranean cold-water corals.
Global environmental changes, including ocean acidification, have been identified as a major threat to scleractinian corals. General predictions are that ocean acidification will be detrimental to reef growth and that 40 to more than 80 per cent of present-day reefs will decline during the next 50 years. Cold-water corals (CWCs) are thought to be strongly affected by changes in ocean acidification owing to their distribution in deep and/or cold waters, which naturally exhibit a CaCO(3) saturation state lower than in shallow/warm waters. Calcification was measured in three species of Mediterranean cold-water scleractinian corals (Lophelia pertusa, Madrepora oculata and Desmophyllum dianthus) on-board research vessels and soon after collection. Incubations were performed in ambient sea water. The species M. oculata was additionally incubated in sea water reduced or enriched in CO(2). At ambient conditions, calcification rates ranged between -0.01 and 0.23% d(-1). Calcification rates of M. oculata under variable partial pressure of CO(2) (pCO(2)) were the same for ambient and elevated pCO(2) (404 and 867 µatm) with 0.06 ± 0.06% d(-1), while calcification was 0.12 ± 0.06% d(-1) when pCO(2) was reduced to its pre-industrial level (285 µatm). This suggests that present-day CWC calcification in the Mediterranean Sea has already drastically declined (by 50%) as a consequence of anthropogenic-induced ocean acidification.


International symposium on the physicochemical field for genetic activities. Awaji Island, Japan, January 24–26, 2011.
The beautiful resort island of Awaji, Japan was recently host to an international gathering of scientists interested in the physical, biochemical and biological aspects of components of the eukaryotic cell nucleus. The meeting featured speakers from many centers of research excellence in Japan as well as from around the world and was characterized by a unique focus on quantitative methods of analysis as applied to chromatin and chromosomes - termed "The Genofield". The meeting was organized by Professors Yasushi Hiraoka, Hitoshi Kurumizaka and Akatsuki Kimura.


Piloting and validating an innovation to triage patients presenting with cough to community pharmacies in Western Australia.
To develop a tool to assist community pharmacists to triage patients presenting with cough and to validate this against an established cough-specific quality of life (QoL) measure. A decision-support tool, the Pharmacy Cough Assessment Tool (PCAT) was developed with reference to published guidelines and a team of clinical experts. The PCAT was validated against the Leicester Cough Questionnaire (LCQ). It was then administered at four community pharmacies in Perth, Western Australia to assess the scope to recruit and follow up participants, and to estimate the proportion of participants who would be advised to consult a general practitioner (GP). The reported outcomes of the consultations with doctors were also recorded. Ninety-nine subjects were recruited over 12 weeks. Thirty-seven participants were advised to consult a GP for further assessment with reference to the PCAT; seven attended their doctor. The LCQ scores of those referred to their GP were significantly lower, indicating a poorer quality of life (adjusted mean and range 13.16 [11.87, 14.46]; non-referred 15.82 [14.47, 17.18]; P < 0.001). Scores of this magnitude have previously been shown to identify patients with chronic respiratory conditions. A smaller group of participants also had a poor quality of life based on LCQ scores but were not referred to their GP. Of the seven participants who made an appointment with their GP, most were prescribed treatment or referred for investigation. There was no significant difference in LCQ score based on gender, or decision to consult a GP. The PCAT identifies patients with cough who might benefit from medical advice and may feasibly be used as an initial screening tool in the community pharmacy setting.


Epigenetic regulation of macrophage polarization and inflammation by DNA methylation in obesity.
Obesity is associated with increased classically activated M1 adipose tissue macrophages (ATMs) and decreased alternatively activated M2 ATMs, both of which contribute to obesity-induced inflammation and insulin resistance. However, the underlying mechanism remains unclear. We find that inhibiting DNA methylation pharmacologically using 5-aza-2'-deoxycytidine or genetically by DNA methyltransferase 1 (DNMT1) deletion promotes alternative activation and suppresses inflammation in macrophages. Consistently, mice with myeloid DNMT1 deficiency exhibit enhanced macrophage alternative activation, suppressed macrophage inflammation, and are protected from obesity-induced inflammation and insulin resistance. The promoter and 5'-untranslated region of peroxisome proliferator-activated receptor γ1 (PPARγ1) are enriched with CpGs and are epigenetically regulated. The saturated fatty acids stearate and palmitate and the inflammatory cytokine TNF-α significantly increase, whereas the TH2 cytokine IL-4 significantly decreases PPARγ1 promoter DNA methylation. Accordingly, inhibiting PPARγ1 promoter DNA methylation pharmacologically using 5-aza-2'-deoxycytidine or genetically by DNMT1 deletion promotes macrophage alternative activation. Our data therefore establish DNA hypermethylation at the PPARγ1 promoter induced by obesity-related factors as a critical determinant of ATM proinflammatory activation and inflammation, which contributes to insulin resistance in obesity.


Improved flow cytometric assessment reveals distinct microvesicle (cell-derived microparticle) signatures in joint diseases.
Microvesicles (MVs), earlier referred to as microparticles, represent a major type of extracellular vesicles currently considered as novel biomarkers in various clinical settings such as autoimmune disorders. However, the analysis of MVs in body fluids has not been fully standardized yet, and there are numerous pitfalls that hinder the correct assessment of these structures. In this study, we analyzed synovial fluid (SF) samples of patients with osteoarthritis (OA), rheumatoid arthritis (RA) and juvenile idiopathic arthritis (JIA). To assess factors that may confound MV detection in joint diseases, we used electron microscopy (EM), Nanoparticle Tracking Analysis (NTA) and mass spectrometry (MS). For flow cytometry, a method commonly used for phenotyping and enumeration of MVs, we combined recent advances in the field, and used a novel approach of differential detergent lysis for the exclusion of MV-mimicking non-vesicular signals. EM and NTA showed that substantial amounts of particles other than MVs were present in SF samples. Beyond known MV-associated proteins, MS analysis also revealed abundant plasma- and immune complex-related proteins in MV preparations. Applying improved flow cytometric analysis, we demonstrate for the first time that CD3(+) and CD8(+) T-cell derived SF MVs are highly elevated in patients with RA compared to OA patients (p=0.027 and p=0.009, respectively, after Bonferroni corrections). In JIA, we identified reduced numbers of B cell-derived MVs (p=0.009, after Bonferroni correction). Our results suggest that improved flow cytometric assessment of MVs facilitates the detection of previously unrecognized disease-associated vesicular signatures.


Elevated pCO2 causes a shift towards more toxic microcystin variants in nitrogen-limited Microcystis aeruginosa.
Elevated pCO2 may promote phytoplankton growth, and potentially alleviate carbon limitation during dense blooms. Under nitrogen-limited conditions, elevated pCO2 may furthermore alter the phytoplankton carbon-nitrogen (C:N) balance and thereby the synthesis of secondary metabolites, such as cyanobacterial toxins. A common group of these toxins are the microcystins, with variants that differ not only in C:N stoichiometry, but also in toxicity. Here, we hypothesized that elevated pCO2 will increase the cellular C:N ratios of cyanobacteria, thereby promoting the more toxic microcystin variants with higher C:N ratios. To test this hypothesis, we performed chemostat experiments under nitrogen-limited conditions, exposing three Microcystis aeruginosa strains to two pCO2 treatments: 400 and 1200 μatm. Biomass, cellular C:N ratios and total microcystin contents at steady state remained largely unaltered in all three strains. Across strains and treatments, however, cellular microcystin content decreased with increasing cellular C:N ratios, suggesting a general stoichiometric regulation. Furthermore, as predicted, microcystin variants with higher C:N ratios generally increased with elevated pCO2, while the variant with a low C:N ratio decreased. Thus, elevated pCO2 under nitrogen-limited conditions may shift the cellular microcystin composition towards the more toxic variants. Such CO2-driven changes may have consequences for the toxicity of Microcystis blooms.


Linker optimization and activity validation of a cell surface vimentin targeted homo-dimeric peptoid antagonist for lung cancer stem cells.
Epithelial-to-mesenchymal transition (EMT) endows epithelia-derived cancer cells with properties of stem cells that govern cancer invasion and metastasis. Vimentin is one of the best studied EMT markers and recent reports indicate that vimentin interestingly translocated onto cell surface under various tumor conditions. We recently reported a cell surface vimentin (CSV) specific peptoid antagonist named JM3A. We now investigated the selective antagonist activity of the optimized homo-dimeric version of JM3A, JM3A-L2D on stem-like cancer cells or cancer stem cells (CSCs) over normal cells in non-small cell lung cancer (NSCLC). Homo-dimerization of JM3A provided the avidity effect and improved the biological activity compared to the monomeric version. We first optimized the central linker length of the dimer by designing seven JM3A derivatives with varying linker lengths/types and evaluated the anti-cancer activity using the standard MTS cell viability assay. The most optimized derivative contains a central lysine linker and two glycines, named JM3A-L2D, which displayed 100 nM vimentin binding affinity (Kd) with an anti-cancer activity (IC50) of 6.7 µM on H1299 NSCLC cells. This is a 190-fold improvement in binding over the original JM3A. JM3A-L2D exhibited better potency on high vimentin-expressing NSCLC cells (H1299 and H460) compared to low vimentin-expressing NSCLC cells (H2122). No activity was observed on normal bronchial HBEC3-KT cells. The anti-CSC activity of JM3A-L2D was evaluated using the standard colony formation assay and JM3A-L2D disrupted the colony formation with IC50 ∼ 400 nM. In addition, JM3A-L2D inhibited cell migration activity at IC50 ∼ 2 µM, assessed via wound healing assay. The underlying mechanism of action seems to be the induction of apoptosis by JM3A-L2D on high-vimentin expressing H1229 and H460 NSCLC cells. Our optimized highly CSV selective peptoid has the potential to be developed as an anti-cancer drug candidate, especially considering the high serum stability and economical synthesis of peptoids.


Fixed vs random effects meta-analysis in rare event studies: the rosiglitazone link with myocardial infarction and cardiac death.
Meta-analyses can be powerful tools to combine the results of randomized clinical trials and observational studies to make consensus inferences about a medical issue. It will be demonstrated that a common practice of testing for homogeneity of effect size, and acting upon the inference to decide between fixed vs random effects, can lead to potentially misleading results. A by-product of this paper is a new ratio estimator approach to random effects meta-analysis of a large set of studies with low event rates. As a case study, we shall use the recent Rosiglitazone example, where diagnostic testing failed to reject homogeneity, leading the investigators to use fixed effects. The results for the fixed and random effects analyses are discordant. In the fixed (random) effects analysis, the p-values for myocardial infarction were 0.03 (0.11) while those for cardiac death were 0.06 (0.0017). Had the fixed effects analysis controlled the study error for multiple testing via a Bonferonni correction, the joint 95+ per cent confidence rectangle for the two outcomes would have included odds ratios of (1.0, 1.0). For the Rosiglitazone example, random effects analysis, where all studies receive the same weight, is the superior choice over fixed effects, where two large studies dominate.


Treatment of a Class II Patient with Four Premolar Extractions and Driftodontics in the Lower Jaw.
The clinical examination of an 11.10-year-old girl in permanent dentition showed severe crowding, chewing problems and her right maxillary lateral incisor was in palatal position. The molars were in Class II relationship on both sides. An extraction treatment was planned. Upper second premolars and lower first premolars were extracted. Lower crowding was eliminated using physiological tooth movement called "driftodontics". Slots sized 0.022 inches and self-ligating brackets were used. The patient also used a high-pull headgear for anchorage and vertical control reasons for a short period. During the correction of the lateral crossbite, a posterior bite plate was used for anterior disarticulation and anterior bite ramps were used at the last stages of the treatment to ease the correction of the curve of Spee. Class I molar and canine relationships and normal overjet and overbite achieved with good alignment of both arches and dental midlines coincided with the facial midline. The treatment results were stable after 2 years of the retention period.


Insulin increases FA uptake and esterification but reduces lipid utilization in isolated contracting muscle.
We examined the effect of insulin on the synthesis and degradation of muscle lipid pools [phospholipid (PL), diacylglycerol (DG), triacylglycerol (TG)] and palmitate oxidation in isolated resting and contracting (20 tetani/min) soleus muscles. Lipid metabolism was monitored using the previously defined pulse-chase procedure. At rest, insulin significantly increased total palmitate uptake into soleus muscle (+49%, P < 0.05), corresponding to enhanced DG (+60%, P < 0.05) and TG (+61%, P < 0.05) esterification, but blunted palmitate oxidation (-38%, P < 0.05) and TG hydrolysis (-34%, P < 0.05). During muscle contraction, when total palmitate uptake was increased, insulin further enhanced uptake (+21%, P < 0.05) and esterification of fatty acids (FA) to PL (+73%, P < 0.05), DG (+19%, P < 0.05), and TG (+161%, P < 0.01). Despite a profound shift in the relative partitioning of FA away from esterification and toward oxidation during contraction, the increase in palmitate oxidation and TG hydrolysis was significantly blunted by insulin [oxidation, -24% (P = 0.05); hydrolysis, -83% (P < 0.01)]. The effects of insulin on FA esterification (stimulation) and oxidation (inhibition) during contraction were reduced in the presence of the phosphatidylinositol 3-kinase inhibitor LY-294002. In summary, the effects of insulin and contraction on palmitate uptake and esterification are additive, while insulin opposes the stimulatory effect of contraction on FA oxidation and TG hydrolysis. Insulin's modulatory effects on muscle FA metabolism during contraction are mediated at least in part through phosphatidylinositol 3-kinase.


The origin and antiquity of syphilis revisited: an appraisal of Old World pre-Columbian evidence for treponemal infection.
For nearly 500 years, scholars have argued about the origin and antiquity of syphilis. Did Columbus bring the disease from the New World to the Old World? Or did syphilis exist in the Old World before 1493? Here, we evaluate all 54 published reports of pre-Columbian, Old World treponemal disease using a standardized, systematic approach. The certainty of diagnosis and dating of each case is considered, and novel information pertinent to the dating of these cases, including radiocarbon dates, is presented. Among the reports, we did not find a single case of Old World treponemal disease that has both a certain diagnosis and a secure pre-Columbian date. We also demonstrate that many of the reports use nonspecific indicators to diagnose treponemal disease, do not provide adequate information about the methods used to date specimens, and do not include high-quality photographs of the lesions of interest. Thus, despite an increasing number of published reports of pre-Columbian treponemal infection, it appears that solid evidence supporting an Old World origin for the disease remains absent.


Protective effect of Phellinus linteus polysaccharide extracts against thioacetamide-induced liver fibrosis in rats: a proteomics analysis.
The hepatoprotective potential of Phellinus linteus polysaccharide (PLP) extracts has been described. However, the molecular mechanism of PLP for the inhibition of liver fibrosis is unclear. This study aims to investigate the molecular protein signatures involved in the hepatoprotective mechanisms of PLP via a proteomics approach using a thioacetamide (TAA)-induced liver fibrosis rat model. Male Sprague-Dawley rats were divided into three groups of six as follows: Normal group; TAA group, in which rats received TAA only; and PLP group, in which rats received PLP and TAA. Liver fibrosis was induced in the rats by repeated intraperitoneal injections of TAA at a dose of 200 mg/kg body weight twice a week for 4 weeks. PLP was given orally at a dose of 50 mg/kg body weight twice a day from the beginning of the TAA treatment until the end of the experiment. The development of liver cirrhosis was verified by histological examination. Liver proteomes were established by two-dimensional gel electrophoresis. Proteins with significantly altered expression levels were identified by matrix-assisted laser desorption/ionization-time of flight/time of flight mass spectrometry and the differentially expressed proteins were validated by immunohistochemical staining and reverse transcription polymerase chain reaction. Histological staining showed a remarkable reduction in liver fibrosis in the rats with PLP treatment. A total of 13 differentially expressed proteins including actin, tubulin alpha-1C chain, preprohaptoglobin, hemopexin, galectin-5, glutathione S-transferase alpha-4 (GSTA4), branched chain keto acid dehydrogenase hterotetrameric E1 subunit alpha (BCKDHA), glutathione S-transferase mu (GSTmu); glyceraldehyde-3-phosphate dehydrogenase (GAPDH); thiosulfate sulfurtransferase (TFT); betaine-homocysteine S-methyltransferase 1 (BHMT1); quinoid dihydropteridine reductase (QDPR); ribonuclease UK114 were observed between the TAA and PLP groups. These proteins are involved in oxidative stress, heme and iron metabolism, cysteine metabolism, and branched-chain amino acid catabolism. The proteomics data indicate that P. linteus may be protective against TAA-induced liver fibrosis via regulation of oxidative stress pathways, heat shock pathways, and metabolic pathways for amino acids and nucleic acids.


Circadian change in bronchial responsiveness and airflow obstruction in asthmatic children.
To throw light on the question of whether the increase in bronchial responsiveness seen during the night is due to increased airflow obstruction, nine asthmatic children with increased airflow obstruction at night (group 1) were compared with nine without (group 2). The mean fall in forced expiratory volume in one second (FEV1) between 16.00 and 04.00 hours was 21.9% in group 1 and 2.3% in group 2. Selection of patients was based on the amplitude of change in peak expiratory flow (PEF) measured every four hours for three consecutive days at home. The study was performed in hospital on four consecutive days. Medication was withheld for three days before and during the measurements at home and in hospital. On the first day in hospital (day 4) FEV1 was measured every four hours for 24 hours. On day 6 inhaled histamine provocation tests were performed at the same times as the FEV1 measurements on day 4. Both groups showed a nocturnal fall in the provocative dose of histamine causing a 20% fall in FEV1 (PC20). The mean change in histamine PC20 from 16.00 to 04.00 hours was 1.1 doubling doses of histamine in group 1 and 1.5 doubling doses in group 2. The results indicate that the increase in nocturnal bronchial responsiveness that occurs at night is not due to an increase in airflow obstruction.


Capital Femoral Epiphyseal Extension May Confer Physeal Stability in Slipped Capital Femoral Epiphysis.
Factors including obesity and morphologic parameters around the hip that increase physeal stress are associated with an increased risk of slipped capital femoral epiphysis (SCFE). Recent evidence suggests that superior epiphyseal extension may confer stability to the physis and help protect against SCFE. The purpose of this study is to investigate the relationship between epiphyseal extension and SCFE using an age-matched and sex-matched cohort study. We generated 2 separate cohorts for comparison: 89 patients with unilateral SCFE and 89 healthy subjects with no evidence of hip disease or deformity. We utilized the anterior-posterior and lateral films of the hip to measure the Southwick angle and the epiphyseal extension ratio (EER), defined as the ratio of extension of the capital femoral epiphysis down the femoral neck relative to the diameter of the femoral head. We then compared these measurements between cohorts and in subgroup analysis based on slip stability and whether subjects progressed to a contralateral slip. The SCFE cohort demonstrated a decreased superior epiphyseal extension ratio compared with control (superior EER 0.71 vs. 0.68, P=0.002). There was also a significant downward trend in superior EER from the control subjects (0.71±0.07) to the stable slips (0.69±0.06) to the unstable slips (0.65±0.04) with an overall difference between the groups (P=0.001). Eighteen of 44 (41%) subjects with unilateral stable slips and at least 6 months of follow-up went on to develop SCFE of the contralateral limb. The subjects who developed contralateral slips were younger (11.6±1.2 vs. 12.7±1.4 y, P=0.008); however, there was no difference in superior or anterior epiphyseal extension (P=0.75 and 0.23, respectively). There was no significant linear correlation between Southwick angle and superior or anterior EER (r=0.13 and 0.17, respectively, P>0.05 for both). Increasing capital femoral epiphyseal extension may confer physeal stability in the setting of SCFE. We propose that this epiphyseal extension reflects an adaptive response to limit physeal stress and reduce the risk for progression to SCFE. Level III-prognostic study.


The benefits of yoga for rheumatoid arthritis: results of a preliminary, structured 8-week program.
The aim of this study was to measure the effects of a bi-weekly Raj yoga program on rheumatoid arthritis (RA) disease activity. Subjects were recruited from among RA patients in Dubai, United Arab Emirates by email invitations of the RA database. Demographic data, disease activity indices, health assessment questionnaire (HAQ), and quality of life (QOL) by SF-36 were documented at enrollment and after completion of 12 sessions of Raj yoga. A total of 47 patients were enrolled: 26 yoga and 21 controls. Baseline demographics were similar in both groups. Patients who underwent yoga had statistically significant improvements in DAS28 and HAQ, but not QOL. Our pilot study of 12 sessions of yoga for RA was able to demonstrate statistically significant improvements in RA disease parameters. We believe that a longer duration of treatment could result in more significant improvements.


Self-perceived acute psychological stress and risk of mortality, recurrence and disability after stroke: Mashhad Stroke Incidence Study.
This longitudinal study was designed to evaluate the association between acute pre-stroke stress and the severity stroke and its outcomes including mortality, recurrence, disability and functional dependency. Patients with first-ever stroke (FES) were recruited from the Mashhad Stroke Incidence Study. Patients were asked about any acute severe pre-stroke stress in the 2 weeks prior to index stroke. Disability and functional disability were defined using modified the Rankin Scale and Barthel Index, respectively. We used logistic and ordinal regression tests to assess the association between acute pre-stroke stress and study outcomes. Among 624 patients with FES, 169 reported acute pre-stroke stress. Patients with acute pre-stroke stress were younger than those without stress (60.7 ± 14.4 vs. 66.2 ± 14.7; p < 0.001). The frequency of traditional vascular risk factors was not different in patients with and without acute pre-stroke stress. We did not find any association between acute pre-stroke stress and stroke outcomes. Although acute stress was common in our cohort, our results did not support an association between acute pre-stroke stress and the severity of stroke at admission and long-term stroke outcomes.


Isotope effect in superconducting n-doped SrTiO3.
We report the influence on the superconducting critical temperature Tc in doped SrTiO3 of the substitution of the natural 16O atoms by the heavier isotope 18O. We observe that for a wide range of doping this substitution causes a strong (~50%) enhancement of Tc. Also the magnetic critical field Hc2 is increased by a factor ~2. Such a strong impact on Tc and Hc2, with a sign opposite to conventional superconductors, is unprecedented. The observed effect could be the consequence of strong coupling of the doped electrons to lattice vibrations (phonons), a notion which finds support in numerous optical and photo-emission studies. The unusually large size of the observed isotope effect supports a recent model for superconductivity in these materials based on strong coupling to the ferroelectric soft modes of SrTiO3.


Nutrition-associated problems facing elite level three-day eventing horses.
The main goal of feeding elite 3-day event horses is to deliver nutrients in optimal amounts to allow the horse to maximize its health and performance. However, improper nutritional management and/or physiological stressors related to intense training and competition may increase the risk of nutrition-associated disorders in these horses. An understanding of the nutrition-associated problems contributing to poor performance is critical to the health and welfare of the horse. To characterize the nutrition-associated problems affecting top level 3-day event horses during 2008. Contact information for riders competing in the 2 highest levels of 3-day eventing in 2008 was obtained from the United States Eventing Association. A survey containing 10 questions pertaining to participant demographics and nutrition-associated problems experienced by their horses was mailed and e-mailed to the 81 individuals fitting our criteria of living in USA and Canada. Data was collected in April and May 2009. Twenty-nine of 81 riders completed the survey (35.8%). Respondents rode a total of 45 horses in top level 3-day events in 2008. The top 5 nutrition-associated problems that horses faced at a significantly higher level than the other problems (P < 0.0001) were gastric ulcers (42.2%), joint problems (37.7%), decreased appetite (31.1%), weight loss (31.1%) and hyperexcitability (22.2%). There was no significant difference in frequency of problems between home and competition (P = 0.22). Horses competing at a high level of 3-day eventing in 2008 were at risk of reduced performance given the significant rate of gastric ulcers, decreased appetite and weight loss. Research addressing specific causes of and/or feeding management changes that would reduce the incidence of these problems in these horses is needed to ensure optimal health and performance.


eHealth interventions for people with chronic kidney disease.
Chronic kidney disease (CKD) is associated with high morbidity and death, which increases as CKD progresses to end-stage kidney disease (ESKD). There has been increasing interest in developing innovative, effective and cost-efficient methods to engage with patient populations and improve health behaviours and outcomes. Worldwide there has been a tremendous increase in the use of technologies, with increasing interest in using eHealth interventions to improve patient access to relevant health information, enhance the quality of healthcare and encourage the adoption of healthy behaviours. This review aims to evaluate the benefits and harms of using eHealth interventions to change health behaviours in people with CKD. We searched the Cochrane Kidney and Transplant Register of Studies up to 14 January 2019 through contact with the Information Specialist using search terms relevant to this review. Studies in the Register are identified through searches of CENTRAL, MEDLINE, and EMBASE, conference proceedings, the International Clinical Trials Register (ICTRP) Search Portal and ClinicalTrials.gov. Randomised controlled trials (RCTs) and quasi-RCTs using an eHealth intervention to promote behaviour change in people with CKD were included. There were no restrictions on outcomes, language or publication type. Two authors independently assessed trial eligibility, extracted data and assessed the risk of bias. The certainty of the evidence was assessed using GRADE. We included 43 studies with 6617 participants that evaluated the impact of an eHealth intervention in people with CKD. Included studies were heterogeneous in terms of eHealth modalities employed, type of intervention, CKD population studied and outcomes assessed. The majority of studies (39 studies) were conducted in an adult population, with 16 studies (37%) conducted in those on dialysis, 11 studies (26%) in the pre-dialysis population, 15 studies (35%) in transplant recipients and 1 studies (2%) in transplant candidates We identified six different eHealth modalities including: Telehealth; mobile or tablet application; text or email messages; electronic monitors; internet/websites; and video or DVD. Three studies used a combination of eHealth interventions. Interventions were categorised into six types: educational; reminder systems; self-monitoring; behavioural counselling; clinical decision-aid; and mixed intervention types. We identified 98 outcomes, which were categorised into nine domains: blood pressure (9 studies); biochemical parameters (6 studies); clinical end-points (16 studies); dietary intake (3 studies); quality of life (9 studies); medication adherence (10 studies); behaviour (7 studies); physical activity (1 study); and cost-effectiveness (7 studies).Only three outcomes could be meta-analysed as there was substantial heterogeneity with respect to study population and eHealth modalities utilised. There was found to be a reduction in interdialytic weight gain of 0.13kg (4 studies, 335 participants: MD -0.13, 95% CI -0.28 to 0.01; I2 = 0%) and a reduction in dietary sodium intake of 197 mg/day (2 studies, 181 participants: MD -197, 95% CI -540.7 to 146.8; I2 = 0%). Both dietary sodium and fluid management outcomes were graded as being of low evidence due to high or unclear risk of bias and indirectness (interdialytic weight gain) and high or unclear risk of bias and imprecision (dietary sodium intake). Three studies reported death (2799 participants, 146 events), with 45 deaths/1000 cases compared to standard care of 61 deaths/1000 cases (RR 0.74, CI 0.53 to 1.03; P = 0.08). We are uncertain whether using eHealth interventions, in addition to usual care, impact on the number of deaths as the certainty of this evidence was graded as low due to high or unclear risk of bias, indirectness and imprecision. eHealth interventions may improve the management of dietary sodium intake and fluid management. However, overall these data suggest that current evidence for the use of eHealth interventions in the CKD population is of low quality, with uncertain effects due to methodological limitations and heterogeneity of eHealth modalities and intervention types. Our review has highlighted the need for robust, high quality research that reports a core (minimum) data set to enable meaningful evaluation of the literature.


The Great American Recession and forgone healthcare: Do widened disparities between African-Americans and Whites remain?
During the Great Recession in America, African-Americans opted to forgo healthcare more than other racial/ethnic groups. It is not understood whether disparities in forgone care returned to pre-recession levels. Understanding healthcare utilization patterns is important for informing subsequent efforts to decrease healthcare disparities. Therefore, we examined changes in racial disparities in forgone care before, during, and after the Great Recession. Data were pooled from the 2006-2013 National Health Interview Survey. Forgone medical, mental, and prescription care due to affordability were assessed among African-Americans and Whites. Time periods were classified as: pre-recession (May 2006-November 2007), early recession (December 2007-November 2008), late recession (December 2008-May 2010) and post-recession (June 2010-December 2013). Multivariable logistic regressions of race, interacted with time periods, were used to identify disparities in forgone care controlling for other demographics, health insurance coverage, and having a usual place for medical care across time periods. Adjusted Wald tests were performed to identify significant changes in disparities across time periods. The sample consisted of 110,746 adults. African-Americans were more likely to forgo medical care during the post- recession compared to Whites (OR = 1.16, CI = 1.06, 1.26); changes in foregone medical care disparities were significant in that they increased in the post-recession period compared to the pre-recession (OR = 1.17, CI = 1.08, 1.28 and OR = 0.89, CI = 0.77, 1.04, respectively, adjusted Wald Test p-value < 0.01). No changes in disparities were seen in prescription and mental forgone care. A persistent increase in forgone medical care disparities existed among African-Americans compared to Whites post-Great Recession and may be a result of outstanding issues related to healthcare access, cost, and quality. While health insurance is an important component of access to care, it alone should not be expected to remove these disparities due to other financial constraints. Additional strategies are necessary to close remaining gaps in care widened by the Great Recession.


Correlation of intercentromeric distance, mosaicism, and sexual phenotype: molecular localization of breakpoints in isodicentric Y chromosomes.
Isodicentric chromosomes are among the structural abnormalities of the Y chromosome that are commonly identified in patients. The simultaneous 45,X cell line that is generated in cell division due to instability of the isodicentric Y chromosome [idic(Y)] has long been hypothesized to explain the variable sexual development of these patients, although gonads have been studied in only a subset of cases. We report here on the molecular localization of breakpoints in ten patients with an idic(Y). Breakpoints were mapped by FISH using BACs; gonads and fibroblasts were also analyzed when possible to evaluate the level of mosaicism. First, we demonstrate great tissue variability in the distribution of idic(Y). Second, palindromes and direct repeats were near the breakpoint of several idic(Y), suggesting that these sequences play a role in the formation of idic(Y). Finally, our data suggest that intercentromeric distance has a negative influence on the stability of idic(Y), as a greater proportion of cells with breakage or loss of the idic(Y) were found in idic(Y) with a greater intercentromeric distance. Females had a significantly greater intercentromeric distance on their idic(Y) than did males. In conclusion, our study indicates that the Y chromosome contains sequences that are more prone to formation of isodicentric chromosomes. We also demonstrate that patients with an intercentromeric distance greater than 20 Mb on their idic(Y) are at increased risk of having a female sexual phenotype.


Inhibition of membrane-type 1 matrix metalloproteinase by hydroxamate inhibitors: an examination of the subsite pocket.
The membrane-type 1 matrix metalloproteinase (MT1-MMP) has been reported to mediate the activation of pro-gelatinase A (proMMP-2), which is associated with tumor proliferation and metastasis. MT1-MMP can also digest extracellular matrix (ECM) such as interstitial collagens, gelatin, and proteoglycan and thus may play an important role in pathophysiological digestion of ECM. We studied the inhibitory effect of various hydroxamate MMP inhibitors, including known inhibitors such as BB-94, BB-2516, GM6001, and Ro31-9790, on a deletion mutant of MT1-MMP lacking the transmembrane domain (DeltaMT1) to further characterize the enzyme and develop a selective inhibitor for MT1-MMP. The evaluation of the inhibitory activities of various hydroxamates reveals general structural profiles affecting selectivities toward MMPs. In particular, a longer side chain at the P1' position is preferable for the binding to MMP-2, -3, and -9 and MT1-MMP. For the P2' position, an alpha-branched alkyl group is critical for the binding toward DeltaMT1, while the introduction of a bulky group at the alpha-position of hydroxamic acid seems to diminish the activity against DeltaMT1. Summation of the data on the sensitivity of DeltaMT1 to various hydroxamate inhibitors indicates that (1) the volume of the S1' subsite of DeltaMT1 is similar to that of MMP-2, -3, and -9, which is bigger than that of MMP-1, and (2) the S1 and S2' subsites are narrower than those in other MMPs. On the basis of these results, the hydroxamates with a P1' phenylpropyl and P2' alpha-branched alkyl group were synthesized and evaluated for inhibitory activity. These inhibitors (1h,i) showed strong activity against DeltaMT1 over MMP-1, but no selectivity between DeltaMT1 and MMP-9. These results are explained using molecular modeling studies conducted on MT1-MMP.


First Report of Cercospora apii Leaf Spot on Capsicum chinense in Brazil.
In Brazil, Capsicum chinense Jacq. is the predominant species of commercial hot peppers because of its popular citrus-like aroma and adaptability to different soils and climates (4). In June 2010, 30 samples of C. chinense with severe leaf spot were collected from a field in the city of Viçosa, state of Minas Gerais, Brazil. Symptoms were observed on leaves, calyxes, fruits, and stems on most of the plants found in the area. On leaves, symptoms included amphigenous lesions that were initially circular to ellipsoid, 1 to 5 mm in diameter, whitish to tan in the center, and surrounded by a dark brown or reddish purple border. Lesions coalesce and turned necrotic with age. A fungus isolated from the lesions matched well with the description of Cercospora apii Fresen. It formed erumpent stromata that were dark brown and spherical to irregular; fascicule conidiophores were clear brown or pale, straight or curved, unbranched, geniculate, 22.5 to 80 × 5 to 7.5 μm, 0 to 3 septate, subtruncate apex; and conidia were solitary, hyaline to subhyaline, filiform, base truncate, tip acute, straight to curved, 12.5 to 140 × 3.5 to 5 μm, and 0 to 11 septate (1,2). A sample was deposited in the herbarium of the Universidade Federal de Viçosa, Minas Gerais, Brazil (VIC 31415). Identity was confirmed by amplifying part of the calmodulin gene with species-specific primers CercoCal-apii and CercoCal-R (3) of fungal DNA from a single-spore culture. In amplification reaction, initial denaturation step was done at 94°C for 5 min, followed by 40 cycles of denaturation at 94°C (30 s), annealing at 56°C (30 s), and elongation at 72°C (30 s). Primers CercoCal-apii and CercoCal-R amplified a single DNA product of 176 bp, and coupled with the morphological characteristics, confirmed the identity of the fungus as Cercospora apii. To check pathogenicity, a 6-mm-diameter plug of the isolate was removed from the expanding edge of a 21-day-old culture grown on potato dextrose agar (PDA) and placed in contact with the adaxial face of the leaves of 8-week-old C. chinense grown in 2-liter plastic pots with soil substrate. Six plants, one per pot, were inoculated with the isolate and six plants were inoculated with the fungus-free PDA plug. Inoculated plants were maintained in a moist chamber for 24 h and then subsequently kept in a greenhouse at 26°C. Leaf spot was observed in all inoculated plants 15 days after inoculation and symptoms were similar to those expressed in the field. The fungus was reisolated from the inoculated plants and matched well with the description of Cercospora apii. All fungus-free PDA inoculated plants remained healthy. Cercospora apii comprises a complex of 281 morphologically indistinguishable species that can infect an extremely wide host range (2). To our knowledge, this pathogen has the potential to cause significant damage to the hot pepper industry of Brazil. References: (1) C. Chupp. A Monograph of the Fungus Cercospora. Cornell University Press, Ithaca, NY, 1954. (2) P. W. Crous and U. Braun. CBS Biodivers. Ser. 1:1, 2003. (3) M. Groenewald et al. Phytopathology 95:951, 2005. (4) S. D. Lannes et al. Sci. Hortic. 112:266, 2007.


A review of trabectedin (ET-743): a unique mechanism of action.
Trabectedin (ET-743) is a marine alkaloid isolated from the Caribbean tunicate Ecteinascidia turbinata, with a chemical structure characterized by three fused tetrahydroisoquinoline rings. Two of these rings (subunits A and B) provide the framework for covalent interaction with the minor groove of the DNA double helix, whereas the third ring (subunit C) protrudes from the DNA duplex, apparently allowing interactions with adjacent nuclear proteins. The compound's chemical interactions trigger a cascade of events that interfere with several transcription factors, DNA binding proteins, and DNA repair pathways, likely to be different from other DNA-interacting agents. Trabectedin also causes modulation of the production of cytokines and chemokines by tumor and normal cells, suggesting that the antitumor activity could also be ascribed to changes in the tumor microenvironment. The promising data on the combination of trabectedin with other anticancer agents, observed in preclinical systems, have prompted several clinical studies that are currently ongoing. One of these combinations (trabectedin-pegylated liposomal doxorubicin) was recently authorized by the European Commission for the treatment of patients with relapsed platinum-sensitive ovarian cancer.


Activity of wild Japanese macaques in Yakushima revealed by camera trapping: Patterns with respect to season, daily period and rainfall.
Animals are subject to various scales of temporal environmental fluctuations, among which daily and seasonal variations are two of the most widespread and significant ones. Many biotic and abiotic factors change temporally, and climatic factors are particularly important because they directly affect the cost of thermoregulation. The purpose of the present study was to determine the activity patterns of wild Japanese macaques (Macaca fuscata) with a special emphasis on the effect of thermal conditions. We set 30 camera traps in the coniferous forest of Yakushima and monitored them for a total of 8658 camera-days between July 2014 and July 2015. Over the one-year period, temperature had a positive effect, and rainfall had a negative effect on the activity of macaques during the day. Capture rate was significantly higher during the time period of one hour after sunrise and during midday. During winter days, macaques concentrated their activity around noon, and activity shifted from the morning toward the afternoon. This could be interpreted as macaques shifting their activity to warmer time periods within a single day. Japanese macaques decreased their activity during the time before sunrise in seasons with lower temperatures. It was beneficial for macaques to be less active during cooler time periods in a cold season. Even small amounts of rainfall negatively affected the activity of Japanese macaques, with capture rates decreasing significantly even when rainfall was only 0.5-1 mm/min. In conclusion, thermal conditions significantly affected the activity of wild Japanese macaques at various time scales.


Chimpanzees in an anthropogenic landscape: Examining food resources across habitat types at Bossou, Guinea, West Africa.
Many primate populations occur outside protected areas in fragmented anthropogenic landscapes. Empirical data on the ecological characteristics that define an anthropogenic landscape are urgently required if conservation initiatives in such environments are to succeed. The main objective of our study was to determine the composition and availability of chimpanzee (Pan troglodytes verus) food resources across fine spatial scales in the anthropogenic landscape of Bossou, Guinea, West Africa. We examined food resources in all habitat types available in the chimpanzees' core area. We surveyed resource composition, structure and heterogeneity (20 m × 20 m quadrats, N = 54) and assessed temporal availability of food from phenology trails (total distance 5951 m; 1073 individual trees) over 1 year (2012-2013). Over half of Bossou consists of regenerating forest and is highly diverse in terms of chimpanzee food species; large fruit bearing trees are rare and confined to primary and riverine forest. Moraceae (mulberries and figs) was the dominant family, trees of which produce drupaceous fruits favored by chimpanzees. The oil palm occurs at high densities throughout and is the only species found in all habitat types except primary forest. Our data suggest that the high densities of oil palm and fig trees, along with abundant terrestrial herbaceous vegetation and cultivars, are able to provide the chimpanzees with widely available resources, compensating for the scarcity of large fruit trees. A significant difference was found between habitat types in stem density/ha and basal area m2 /ha of chimpanzee food species. Secondary, young secondary, and primary forest emerged as the most important habitat types for availability of food tree species. Our study emphasizes the importance of examining ecological characteristics of an anthropogenic landscape as each available habitat type is unlikely to be equally important in terms of spatial and temporal availability of resources. Am. J. Primatol. 78:1237-1249, 2016. © 2016 Wiley Periodicals, Inc.


Self-Assembled Sandwich-like Vanadium Oxide/Graphene Mesoporous Composite as High-Capacity Anode Material for Lithium Ion Batteries.
Sandwich-like V2O5/graphene mesoporous composite has been synthesized by a facile solvothermal approach. The crystalline structure, morphology, and electrochemical performance of the as-prepared materials have been investigated in detail. The results demonstrate that the 30-50 nm V2O5 particles are homogeneously anchored on conducting graphene sheets, which allow the V2O5 nanoparticles to be wired up to a current collector through the underlying conducting graphene layers. As an anode material for lithium ion batteries, the composite exhibits a high reversible capacity of 1006 mAh g(-1) at a current density of 0.5 A g(-1) after 300 cycles. It also exhibits excellent rate performance with a discharge capacity of 500 mAh g(-1) at the current density of 3.0 A g(-1), which is superior to the performance of the vanadium-based materials reported previously. The electrochemical properties demonstrate that the sandwich-like V2O5/graphene mesoporous composite could be a promising candidate material for high-capacity anode in lithium ion batteries.


Handling intra-cluster correlation when analyzing the effects of decision support on health care process measures.
The clinical worksite constitutes a naturally clustered environment, posing challenges in the statistical analysis of quality improvement interventions such as computerized decision support. Ignoring clustering in the analysis may lead to biased effect estimates, underestimating the variance and hence type I errors. This paper presents a secondary analysis on data from a previously published, cluster randomized trial in cardiac rehabilitation. We compared six different statistical analysis methods (weighted and unweighted t-test; adjusted χ2 test; normal and multilevel logistic regression analysis; and generalized estimation equations). There were considerable differences in both point estimates and p-values derived by the methods, and differences were larger with increasing intracluster correlation.


Origin of the TTC values for compounds that are genotoxic and/or carcinogenic and an approach for their re-evaluation.
The threshold of toxicological concern (TTC) approach is a resource-effective de minimis method for the safety assessment of chemicals, based on distributional analysis of the results of a large number of toxicological studies. It is being increasingly used to screen and prioritize substances with low exposure for which there is little or no toxicological information. The first step in the approach is the identification of substances that may be DNA-reactive mutagens, to which the lowest TTC value is applied. This TTC value was based on the analysis of the cancer potency database and involved a number of assumptions that no longer reflect the state-of-the-science and some of which were not as transparent as they could have been. Hence, review and updating of the database is proposed, using inclusion and exclusion criteria reflecting current knowledge. A strategy for the selection of appropriate substances for TTC determination, based on consideration of weight of evidence for genotoxicity and carcinogenicity is outlined. Identification of substances that are carcinogenic by a DNA-reactive mutagenic mode of action and those that clearly act by a non-genotoxic mode of action will enable the protectiveness to be determined of both the TTC for DNA-reactive mutagenicity and that applied by default to substances that may be carcinogenic but are unlikely to be DNA-reactive mutagens (i.e. for Cramer class I-III compounds). Critical to the application of the TTC approach to substances that are likely to be DNA-reactive mutagens is the reliability of the software tools used to identify such compounds. Current methods for this task are reviewed and recommendations made for their application.


Cognitive training for attention-deficit/hyperactivity disorder: meta-analysis of clinical and neuropsychological outcomes from randomized controlled trials.
The authors performed meta-analyses of randomized controlled trials to examine the effects of cognitive training on attention-deficit/hyperactivity disorder (ADHD) symptoms, neuropsychological deficits, and academic skills in children/adolescents with ADHD. The authors searched Pubmed, Ovid, Web of Science, ERIC, and CINAHAL databases through May 18, 2014. Data were aggregated using random-effects models. Studies were evaluated with the Cochrane risk of bias tool. Sixteen of 695 nonduplicate records were analyzed (759 children with ADHD). When all types of training were considered together, there were significant effects on total ADHD (standardized mean difference [SMD] = 0.37, 95% CI = 0.09-0.66) and inattentive symptoms (SMD = 0.47, 95% CI = 0.14-0.80) for reports by raters most proximal to the treatment setting (i.e., typically unblinded). These figures decreased substantially when the outcomes were provided by probably blinded raters (ADHD total: SMD = 0.20, 95% CI = 0.01-0.40; inattention: SMD = 0.32, 95% CI = -0.01 to 0.66). Effects on hyperactivity/impulsivity symptoms were not significant. There were significant effects on laboratory tests of working memory (verbal: SMD = 0.52, 95% CI = 0.24-0.80; visual: SMD = 0.47, 95% CI = 0.23-0.70) and parent ratings of executive function (SMD = 0.35, 95% CI = 0.08-0.61). Effects on academic performance were not statistically significant. There were no effects of working memory training, specifically on ADHD symptoms. Interventions targeting multiple neuropsychological deficits had large effects on ADHD symptoms rated by most proximal assessors (SMD = 0.79, 95% CI = 0.46-1.12). Despite improving working memory performance, cognitive training had limited effects on ADHD symptoms according to assessments based on blinded measures. Approaches targeting multiple neuropsychological processes may optimize the transfer of effects from cognitive deficits to clinical symptoms.


S-nitrosylation is emerging as a specific and fundamental posttranslational protein modification: head-to-head comparison with O-phosphorylation.
Nitric oxide (NO) is a free-radical product of mammalian cell metabolism that plays diverse and important roles in the regulation of cell function. Biological actions of NO arise as a direct consequence of chemical reactions between NO or NO-derived species and protein targets. Reactions of NO with transition metals in target proteins have garnered the most attention to date as the principal mechanism of NO signaling; nonetheless, S-nitrosylation of protein Cys residues is rapidly moving to center stage in importance. In general, however, there has been a delay in adequate appreciation of the role of S-nitrosylation in biological signaling by NO. This lag is attributed to a poor understanding of the basis for selective targeting of NO to particular thiols, and methodological limitations in accurately quantifying this modification--recent breakthroughs in concepts and methods diminish these barriers. Here, we consider the wheres and whys of protein S-nitrosylation and its basis for specificity. Protein S-nitrosylation potentially represents a ubiquitous and fundamental mechanism for posttranslational control of protein activity on a par with that of O-phosphorylation.


A Polymorphic Antioxidant Response Element Links NRF2/sMAF Binding to Enhanced MAPT Expression and Reduced Risk of Parkinsonian Disorders.
The NRF2/sMAF protein complex regulates the oxidative stress response by occupying cis-acting enhancers containing an antioxidant response element (ARE). Integrating genome-wide maps of NRF2/sMAF occupancy with disease-susceptibility loci, we discovered eight polymorphic AREs linked to 14 highly ranked disease-risk SNPs in individuals of European ancestry. Among these SNPs was rs242561, located within a regulatory region of the MAPT gene (encoding microtubule-associated protein Tau). It was consistently occupied by NRF2/sMAF in multiple experiments and its strong-binding allele associated with higher mRNA levels in cell lines and human brain tissue. Induction of MAPT transcription by NRF2 was confirmed using a human neuroblastoma cell line and a Nrf2-deficient mouse model. Most importantly, rs242561 displayed complete linkage disequilibrium with a highly protective allele identified in multiple GWASs of progressive supranuclear palsy, Parkinson's disease, and corticobasal degeneration. These observations suggest a potential role for NRF2/sMAF in tauopathies and a possible role for NRF2 pathway activators in disease prevention.


Charge storage performances and mechanisms of MnO2 nanospheres, nanorods, nanotubes and nanosheets.
Manganese dioxide (MnO2) has been widely used as an active material for high-performance supercapacitors due to its high theoretical capacitance, high cycling stability, low cost, and environmental friendliness. However, the effect of its crystallographic phase on charge storage performances and mechanisms is not yet clear. Herein, MnO2-based supercapacitors with different structures including nanospheres, nanorods, nanotubes, and nanosheets have been fabricated and investigated. Among such structures, δ-MnO2 nanosheets exhibit the highest specific capacitance of 194.3 F g-1 at 1 A g-1 when compared with other phases and shapes. The maximum specific energy of the δ-MnO2 nanosheet supercapacitor is 23.4 W h kg-1 at 971.6 W kg-1 and the maximum specific power is 4009.2 W kg-1 at 15.9 W h kg-1 with a capacity retention of 97% over 15 000 cycles. The δ-MnO2 nanosheet mainly stores charges via a diffusion-controlled mechanism at the scan rates of 10-100 mV s-1, whilst the α-MnO2 with different morphologies including nanospheres, nanorods, and nanotubes store charges via a non-faradaic or non-diffusion controlled process especially at fast scan rates (50-100 mV s-1). Understanding the charge storage performance and mechanism of the MnO2 nanostructures with different crystallographic phases and morphologies may lead to the further development of supercapacitors.


Routes of clearance of radioactive water from the rabbit vitreous.
The movement of water to and from the vitreous across the retina and pigment epithelium is important in relation to an understanding of such conditions as retinal detachment and its surgical cure, central serous retinopathy, and retinal oedema. Experiments were carried out to determine the main routes for removal of water injected into the vitreous and to see if the removal could be explained on the basis of diffusion or whether bulk flow was also implicated. 25 microCi of 3H2O in 25 microliter were injected into a central position in the vitreous humour of living rabbits under general anaesthesia. For 9 animals blood was collected from one of the 4 vortex veins draining the choroid and the radioactivity in the samples measured. For another 6 rabbits similarly injected the radioactivity in samples of aqueous humour was determined. The percentage of injectate recovered from the vortex vein blood ranged between 13% and 38%, mean 25 +/- 3%. The percentage of injectate recovered from the aqueous humour ranged between 1.2% and 5.2%, mean 2.8 +/- 0.6%. Analysis of the time course of isotope activity in the samples revealed a mean transit time from the mid vitreous to the choroid of 32 +/- 2 minutes, and from the mid vitreous to the anterior chamber of 84 +/- 3 minutes. By means of a computer model it was calculated that diffusion alone could effect this transfer; if active transport were involved in the transport of 3H2O to the choroid, this was not a limiting factor under the conditions of the experiment.


Wild tobacco genomes reveal the evolution of nicotine biosynthesis.
Nicotine, the signature alkaloid of Nicotiana species responsible for the addictive properties of human tobacco smoking, functions as a defensive neurotoxin against attacking herbivores. However, the evolution of the genetic features that contributed to the assembly of the nicotine biosynthetic pathway remains unknown. We sequenced and assembled genomes of two wild tobaccos, Nicotiana attenuata (2.5 Gb) and Nicotiana obtusifolia (1.5 Gb), two ecological models for investigating adaptive traits in nature. We show that after the Solanaceae whole-genome triplication event, a repertoire of rapidly expanding transposable elements (TEs) bloated these Nicotiana genomes, promoted expression divergences among duplicated genes, and contributed to the evolution of herbivory-induced signaling and defenses, including nicotine biosynthesis. The biosynthetic machinery that allows for nicotine synthesis in the roots evolved from the stepwise duplications of two ancient primary metabolic pathways: the polyamine and nicotinamide adenine dinucleotide (NAD) pathways. In contrast to the duplication of the polyamine pathway that is shared among several solanaceous genera producing polyamine-derived tropane alkaloids, we found that lineage-specific duplications within the NAD pathway and the evolution of root-specific expression of the duplicated Solanaceae-specific ethylene response factor that activates the expression of all nicotine biosynthetic genes resulted in the innovative and efficient production of nicotine in the genus Nicotiana Transcription factor binding motifs derived from TEs may have contributed to the coexpression of nicotine biosynthetic pathway genes and coordinated the metabolic flux. Together, these results provide evidence that TEs and gene duplications facilitated the emergence of a key metabolic innovation relevant to plant fitness.


Screening for acute confusion in elderly long-term care residents.
While acute confusion (AC) is frequently studied in the hospitalized elderly population, this phenomena has been largely ignored in elders who are residents in long-term care (LTC) facilities. The purpose of this study was to estimate the prevalence of AC in older LTC residents, the antecedent conditions present at the time of the AC event and the recognition rate of AC when assessed by staff nurses in two LTC facilities. This is a descriptive, longitudinal study with a 14 day follow-up period which incorporates a screening algorithm using AC instruments with established psychometric properties. A behavioral symptom instrument was also used in order to classify AC cases into clinical subtypes: hyperkinetic, hypokinetic and mixed. Staff nurses, using traditional assessment techniques and blinded to the case screening algorithm outcome, were asked to randomly evaluate both residents who were "clear" and those experiencing AC. Screening was conducted at both daytime and evening time-points. Of the 37 subjects followed. 15 (40.5%) screened positive for AC. Those AC cases with compromised cortical functioning indicative of Alzheimer's disease (n = 4) were particularly vulnerable to poor fluid intake. High risk medications (n = 7) and urinary tract infections (n = 7) were the most frequent antecedents. Most of the subjects experienced two or more antecedent conditions at the time of their AC. Staff nurses were able to identify 4 (26.7%) of the 15 instrument-positive residents, none of which were of a hypokinetic clinical subtype. These early results suggest a high prevalence of AC among elders in LTC, which may go unrecognized by bedside care providers.


Association of Dairy Product Consumption with Metabolic and Inflammatory Biomarkers in Adolescents: A Cross-Sectional Analysis from the LabMed Study.
This study aimed to investigate the association between dairy product consumption and metabolic and inflammatory biomarkers in Portuguese adolescents, and whether the association differed by weight status. A cross-sectional study was conducted during the school year 2011/2012 with 412 Portuguese adolescents (52.4% girls) in 7th and 10th grade (aged 12 to 18 years old). The World Health Organization cutoffs were used to categorize adolescents as non-overweight (NW) or overweight (OW). Blood samples were collected to analyze C-reactive protein (CRP), interleukin-6 (IL-6), leptin, and adiponectin. Dairy product intake was evaluated using a food frequency questionnaire. Participants were divided by tertiles according to the amount of dairy product consumed. The associations between dairy product consumption with metabolic and inflammatory biomarkers were evaluated using generalized linear regression models with logarithmic link and gamma distribution and adjusted for potential confounders. The majority of adolescents were NW (67.2%). NW adolescents had lower IL-6, CRP, and leptin concentration than their counterparts (p < 0.05, for all comparisons). Higher levels of total dairy product and milk intake were inversely associated with IL-6 (P for trend <0.05, for all) in NW adolescents, but not in OW adolescents. NW adolescents in the second tertile of yogurt consumption had lower level of IL-6 compared to those in the first tertile (p = 0.004). Our results suggest an inverse association between total dairy product and milk intake and serum concentrations of IL-6 only among NW adolescents.


A source of strength and empowerment? An exploration of the influence of disabled children on the lives of their mothers in Karachi, Pakistan.
Previous literature has highlighted marginalisation and stigma of children with disabilities in developing countries, but few studies have explored the central care-giving environment and the relationship of the mother and her child with disabilities in this context. A group of women caring for children with disabilities in a low income community in Karachi, Pakistan was identified for the study. The aims were: (1) to explore the influence children with disabilities have on the daily lives of their mothers, (2) to describe the factors which influence the care-giving capacity of mothers. A participatory qualitative research design was implemented. A women's group of caregivers of children with disabilities was formed. Several different tools were used during the course of the group meetings to facilitate discussion including social mapping. A thematic analysis of issues around care-giving and the relationship between the mother and her child with disabilities was conducted. In a society where women may experience restrictions in freedom of movement and decision making, caring for a child with disabilities enabled women to move beyond traditional boundaries in seeking health and education services for their children. However, the gain in empowerment was counter-acted by a lack of care-giving support, a lack of appropriate services for health, rehabilitation and education of children with disabilities and stigma creating anxiety and stress for women. While children with disabilities do have some positive effects on the lives of their mothers, there are many more factors which create anxiety for this group of mothers (including a lack of care-giving support and stigma). Community-based rehabilitation strategies should consider the care-giving environment of children with disabilities and shift from a child only to family focus. Interventions which support, empower and strengthen the capacity of mothers are essential for the well being of their children with disabilities.


Hydrogen bonding involving side chain exchangeable groups stabilizes amyloid quarternary structure.
The amyloid β-peptide (Aβ) is the major structural component of amyloid fibrils in the plaques of brains of Alzheimer's disease patients. Numerous studies have addressed important aspects of secondary and tertiary structure of fibrils. In electron microscopic images, fibrils often bundle together. The mechanisms which drive the association of protofilaments into bundles of fibrils are not known. We show here that amino acid side chain exchangeable groups like e.g. histidines can provide useful restraints to determine the quarternary assembly of an amyloid fibril. Exchangeable protons are only observable if a side chain hydrogen bond is formed and the respective protons are protected from exchange. The method relies on deuteration of the Aβ peptide. Exchangeable deuterons are substituted with protons, before fibril formation is initiated.


Influence of path integration versus environmental orientation on place cell remapping between visually identical environments.
To assess the effects of interactions between angular path integration and visual landmarks on the firing of hippocampal neurons, we recorded from CA1 pyramidal cells as rats foraged in two identical boxes with polarizing internal cues. In the same-orientation condition, following an earlier experiment by Skaggs and McNaughton, the boxes were oriented identically and connected by a corridor. In the opposite-orientation condition, the boxes were abutted by rotating them 90 degrees in opposite directions, so that their orientations differed by 180 degrees . After 16-23 days of pretraining on the same-orientation condition, three rats experienced both conditions in counterbalanced order on each of two consecutive days. On the third day they ran two opposite-orientation trials. Although Skaggs and McNaughton observed stable partial "remapping" of place fields, none of the fields in this experiment remapped in the same-orientation condition. In the opposite-orientation condition, place fields in the first box were isomorphic with those in the same-orientation condition, whereas in the second box the rats eventually exhibited completely different fields. The rats differed as to the trial in which this first occurred. Once the second box exhibited different fields, it continued to do so in all subsequent opposite-orientation trials, yet fields remained the same in subsequent same-orientation trials. The results demonstrate that when animals move actively between environments, and are thus potentially able to maintain their inertial angular orientation, discordance between environmental orientation and the rat's idiothetic direction sense can profoundly affect the hippocampal map-either immediately, or as a result of cumulative experience.


Third or greater penetrating keratoplasties: indications, survival, and visual outcomes.
The purposes of this study were to report the indications, graft survival, risk factors for graft failure, and visual outcomes for third or greater penetrating keratoplasties (PKP). Six years of Wills Eye Institute Cornea Service charts (2000-2005) were retrospectively reviewed to identify all patients who had undergone three or more PKPs. Graft survival rates by initial diagnosis and risk factors for graft failure were analyzed. Forty-five patients who had three or more PKPs were identified. The total number of grafts identified in these 45 patients was 152, including 45 third PKPs, 11 fourth, three fifth, two sixth, and one seventh. The most common indication for the initial PKP was pseudophakic bullous keratopathy in 18 of 45 patients (41%) followed by Fuchs dystrophy (seven of 45 patients [16%]) and stromal dystrophies (five of 45 patients [11%]). Approximately half of third grafts (24 of 45 [53%]) and one fourth of fourth grafts (three of 11 [27%]) survived at the last follow-up visit with a median follow up of 4.3 years for the third grafts and 8.4 years for the fourth grafts. One-, 2-, and 5-year graft survival rates were 89%, 78%, and 53% for the third grafts and 73%, 73%, and 64% for the fourth grafts, respectively. The median survival time for the third graft was 12.8 years in Fuchs dystrophy, 5.2 years in herpetic keratitis, 4.0 years in keratoconus, 3.0 years in pseudophakic bullous keratopathy, 2.3 years in iridocorneal endothelial syndrome, and 2.0 years in stromal dystrophies. There is no statistically significant difference between groups (P = 0.46). Risk factor analysis on the third grafts showed that previous glaucoma procedures and corneal neovascularization are statistically significant risk factors for graft failure (P = 0.04 and 0.02, respectively). Over 50% of third and fourth grafts were clear at 5 years postoperatively. Outcomes of third grafts were better in patients with Fuchs dystrophy, keratoconus, and herpetic keratitis. Absence of previous glaucoma surgery and/or corneal neovascularization is associated with better outcomes of multiple PKPs.


Bone fragility and decline in stem cells in prematurely aging DNA repair deficient trichothiodystrophy mice.
Trichothiodystrophy (TTD) is a rare, autosomal recessive nucleotide excision repair (NER) disorder caused by mutations in components of the dual functional NER/basal transcription factor TFIIH. TTD mice, carrying a patient-based point mutation in the Xpd gene, strikingly resemble many features of the human syndrome and exhibit signs of premature aging. To examine to which extent TTD mice resemble the normal process of aging, we thoroughly investigated the bone phenotype. Here, we show that female TTD mice exhibit accelerated bone aging from 39 weeks onwards as well as lack of periosteal apposition leading to reduced bone strength. Before 39 weeks have passed, bones of wild-type and TTD mice are identical excluding a developmental defect. Albeit that bone formation is decreased, osteoblasts in TTD mice retain bone-forming capacity as in vivo PTH treatment leads to increased cortical thickness. In vitro bone marrow cell cultures showed that TTD osteoprogenitors retain the capacity to differentiate into osteoblasts. However, after 13 weeks of age TTD females show decreased bone nodule formation. No increase in bone resorption or the number of osteoclasts was detected. In conclusion, TTD mice show premature bone aging, which is preceded by a decrease in mesenchymal stem cells/osteoprogenitors and a change in systemic factors, identifying DNA damage and repair as key determinants for bone fragility by influencing osteogenesis and bone metabolism.


Generation of a vectorial Mathieu-like hollow beam with a periodically rotated polarization property.
We propose a promising and practical scheme to generate a vectorial Mathieu-like hollow beam (vMHB) by using an axicon optical system including a dual-mode elliptic hollow fiber (EHF), and calculate the intensity and polarization distributions of the beam and its fractional orbital angular momentum (OAM). We also fit its intensity profile and obtain suitable theoretical expressions to describe three-dimensional (3D) propagation characteristics of such a Mathieu-like hollow beam. Our study shows that the generated beam is a vMHB within the propagation distance 1.2-2.2 m, which has an unchanged (diffraction-free) intensity profile and periodically rotated polarization properties as well as an unchanged fractional OAM within 1.2-2.2 m.


Soluble Ig-like transcript 3 inhibits tumor allograft rejection in humanized SCID mice and T cell responses in cancer patients.
Attempts to enhance patients' immune responses to malignancies have been largely unsuccessful. We now describe an immune-escape mechanism mediated by the inhibitory receptor Ig-like transcript 3 (ILT3) that may be responsible for such failures. Using a humanized SCID mouse model, we demonstrate that soluble and membrane ILT3 induce CD8(+) T suppressor cells and prevent rejection of allogeneic tumor transplants. Furthermore, we found that patients with melanoma, and carcinomas of the colon, rectum, and pancreas produce the soluble ILT3 protein, which induces the differentiation of CD8(+) T suppressor cells and impairs T cell responses in MLC. These responses are restored by anti-ILT3 mAb or by depletion of soluble ILT3 from the serum. Immunohistochemical staining of biopsies from the tumors and metastatic lymph nodes suggests that CD68(+) tumor-associated macrophages represent the major source of soluble ILT3. Alternative splicing, resulting in the loss of the ILT3 transmembrane domain, may contribute to the release of ILT3 in the circulation. These data suggest that ILT3 depletion or blockade is crucial to the success of immunotherapy in cancer. In contrast, the inhibitory activity of soluble ILT3 on T cell alloreactivity in vitro and in vivo suggests the potential usefulness of rILT3 for immunosuppressive treatment of allograft recipients or patients with autoimmune diseases.