Apoptosis induced by anthracycline antibiotics in P388 parent and multidrug-resistant cells.
The effect of the topoisomerase II inhibitor doxorubicin and its non-cross-resistant analogue annamycin on DNA degradation and programmed cell death was examined in murine leukemia P388 cells. P388 parental cells exposed to various concentrations of doxorubicin and annamycin for 24 h displayed dose-dependent DNA cleavage: at 1 microM, both doxorubicin and annamycin were effective in inducing DNA breakdown, but at 10 microM, the effect was markedly decreased or totally absent. In multidrug-resistant P388/Dox cells, doxorubicin did not cause DNA cleavage, while 10 microM annamycin had a significant effect. By agarose gel analysis, drug-induced DNA fragmentation showed the characteristic pattern of internucleosomal ladder. Morphologically, P388 cells treated with 1 microM doxorubicin or annamycin for 24 h showed a reduction in cell volume and condensation of nuclear structures. Similar changes were observed in P388/Dox cells exposed to 10 microM annamycin for 24 h but not in cells exposed to 10 microM doxorubicin. Time course studies demonstrated that DNA fragmentation was detected 12 h after incubation with 1 microM doxorubicin or annamycin, while loss of membrane integrity appeared at 24 h, thus indicating that DNA degradation was a preceding event. DNA fragmentation caused by doxorubicin and annamycin was inhibited by the RNA synthesis inhibitor actinomycin D, the protein synthesis inhibitor cycloheximide, and the endonuclease inhibitor aurintricarboxylic acid. Drug-induced cell death was partially prevented by cycloheximide and aurintricarboxylic acid, thus suggesting that the apoptotic process caused by these drugs requires gene expression, synthesis of new proteins, and activation of endogenous nucleases. In contrast, DNA cleavage was not affected by incubating cells with 1 mM ethylene glycol-bis(2-aminoethyl ether)-N,N,N',N'-tetraacetic acid, thus indicating that intracellular calcium depletion does not affect anthracycline-induced apoptosis. The results obtained demonstrate that the cell killing effect of anthracyclines is mediated, at least in part, by the induction of apoptosis.


Inactivation of homologous recombination suppresses defects in topoisomerase III-deficient mutants.
The Saccharomyces cerevisiae TOP3 gene encodes the type IA topoisomerase (Top3p) that is highly conserved in evolution. Deletion of TOP3 leads to a reduction in cell viability, hyper-recombination between repetitive DNA sequences, and abnormalities in both cell cycle progression and responses to DNA damaging agents. Deletion of SGS1, encoding the sole RecQ family helicase in S. cerevisiae, strongly suppresses the phenotypic effects of loss of TOP3 function. Here, we show that many of the adverse phenotypic effects of TOP3 deletion can also be partially alleviated by disruption of homologous recombination (HR) functions. This genetic interaction is seen both in strains deleted for TOP3 and in wild-type strains over-expressing a dominant-negative Top3p mutant form that confers a top3-like phenotype. Moreover, we show that this genetic interaction is conserved in the distantly-related fission yeast, Schizosaccharomyces pombe. Our results implicate topoisomerase III enzymes in recombination repair events required for cellular protection against DNA damaging agents and DNA replication inhibitors.


Sequence features and transcriptional stalling within centromere DNA promote establishment of CENP-A chromatin.
Centromere sequences are not conserved between species, and there is compelling evidence for epigenetic regulation of centromere identity, with location being dictated by the presence of chromatin containing the histone H3 variant CENP-A. Paradoxically, in most organisms CENP-A chromatin generally occurs on particular sequences. To investigate the contribution of primary DNA sequence to establishment of CENP-A chromatin in vivo, we utilised the fission yeast Schizosaccharomyces pombe. CENP-ACnp1 chromatin is normally assembled on ∼10 kb of central domain DNA within these regional centromeres. We demonstrate that overproduction of S. pombe CENP-ACnp1 bypasses the usual requirement for adjacent heterochromatin in establishing CENP-ACnp1 chromatin, and show that central domain DNA is a preferred substrate for de novo establishment of CENP-ACnp1 chromatin. When multimerised, a 2 kb sub-region can establish CENP-ACnp1 chromatin and form functional centromeres. Randomization of the 2 kb sequence to generate a sequence that maintains AT content and predicted nucleosome positioning is unable to establish CENP-ACnp1 chromatin. These analyses indicate that central domain DNA from fission yeast centromeres contains specific information that promotes CENP-ACnp1 incorporation into chromatin. Numerous transcriptional start sites were detected on the forward and reverse strands within the functional 2 kb sub-region and active promoters were identified. RNAPII is enriched on central domain DNA in wild-type cells, but only low levels of transcripts are detected, consistent with RNAPII stalling during transcription of centromeric DNA. Cells lacking factors involved in restarting transcription-TFIIS and Ubp3-assemble CENP-ACnp1 on central domain DNA when CENP-ACnp1 is at wild-type levels, suggesting that persistent stalling of RNAPII on centromere DNA triggers chromatin remodelling events that deposit CENP-ACnp1. Thus, sequence-encoded features of centromeric DNA create an environment of pervasive low quality RNAPII transcription that is an important determinant of CENP-ACnp1 assembly. These observations emphasise roles for both genetic and epigenetic processes in centromere establishment.


Implementation of a reference standard and proficiency testing programme by the World Wide Antimalarial Resistance Network (WWARN).
The Worldwide Antimalarial Resistance Network (WWARN) is a global collaboration to support the objective that anyone affected by malaria receives effective and safe drug treatment. The Pharmacology module aims to inform optimal anti-malarial drug selection. There is an urgent need to define the drug exposure - effect relationship for most anti-malarial drugs. Few anti-malarials have had their therapeutic blood concentration levels defined. One of the main challenges in assessing safety and efficacy data in relation to drug concentrations is the comparability of data generated from different laboratories. To explain differences in anti-malarial pharmacokinetics in studies with different measurement laboratories it is necessary to confirm the accuracy of the assay methods. This requires the establishment of an external quality assurance process to assure results that can be compared. This paper describes this process. The pharmacology module of WWARN has established a quality assurance/quality control (QA/QC) programme consisting of two separate components:1. A proficiency testing programme where blank human plasma spiked with certified reference material (CRM) in different concentrations is sent out to participating bioanalytical laboratories.2. A certified reference standard programme where accurately weighed amounts of certified anti-malarial reference standards, metabolites, and internal standards are sent to participating bioanalytical and in vitro laboratories. The proficiency testing programme is designed as a cooperative effort to help participating laboratories assess their ability to carry out drug analysis, resolve any potential problem areas and to improve their results - and, in so doing, to improve the quality of anti-malarial pharmacokinetic data published and shared with WWARN.By utilizing the same source of standards for all laboratories, it is possible to minimize bias arising from poor quality reference standards. By providing anti-malarial drug standards from a central point, it is possible to lower the cost of these standards.


Chemokine growth-regulated oncogene 1 as a putative biomarker for gastric cancer progression.
Gastric cancer (GC) is a heterogeneous disease that is not well detected by current tumor markers. Identifying molecular markers that can predict the potential for tumor progression is important for appropriate individualized therapy. Using the Cancer Metastasis Research Center microarray database (17K cDNA microarray), we identified genes that were differentially expressed between 96 cancer and 98 normal gastric tissues using significant analysis of microarrays. From these, we selected genes that were overexpressed more than twofold in tumor tissues that encode secreted proteins. The selected genes were validated with ELISA using the sera of 96 GC patients and 48 healthy donors. Our first round of selection included 6510 genes that were differentially expressed between 96 cancer and 98 normal gastric tissues with a minimal false discovery rate of 0.005%. Out of those genes, we picked 386 that encoded secreted proteins based on the SOURCE database. Of these genes, we focused on 55 that were overexpressed more than twofold in GC compared to normal tissues. With Ingenuity Pathway Analysis, we found 34 genes related to cancer. One in particular, chemokine growth-regulated oncogene 1, CXCL1, has been linked to cancer progression in various cancer types, but not yet to GC. Levels of CXCL1 in serum samples of GC patients were significantly higher compared with healthy donors (P < 0.05). Within GC patients, CXCL1 serum levels increased according to tumor stage and lymph node metastasis. The CXCL1 gene appears to be a candidate marker for GC progression.


Dendritic cell maturation with CpG for tumor immunotherapy.
Bacterial DNA has immunostimulatory effects on different types of immune cells such as dendritic cells (DCs). Application of DCs as a cellular adjuvant represents a promising approach in the immunotherapy of infectious disease and cancers. To investigate the effect of tumor antigen pulsed DCs in the presence of CpG-1826 in treatment of a murine model of cancer. WEHI-164 cells (Balb/c derived fibrosarcoma cell line) were injected subcutaneously in the right flank of mice. Bone marrow cells were cultured in the presence of GM-CSF and IL-4. After 5 days, tumor lysate, CpG-1826, and oligodeoxynucleosides, as control, were added to the culture media and incubated for 2 days. Cytokine production in DCs culture media was measured by ELISA. Then DCs were injected subcutaneously around the tumor site in the right flank of mice. Tumor growth rate was monitored in case and control groups. Two weeks after DCs immunotherapy, cytotoxic assay was conducted using various amounts of effector (splenic T cells) and target cells (WEHI-164 or CT26) for 6 h. Immunotherapy with DCs treated with CpG led to a significant increase in the activity of cytotoxic T cells and decreased tumor growth in immunized mice. In the control group which received DCs without CpG treatment, no change in cytotoxic activity and tumor growth rate was detected. The current study suggests that specific anti tumor immune responses can be induced by DCs matured with CpG and proposes CpG usage in DCs targeted clinical strategies.


Surgical Oncology: Evolution of Postoperative Fatigue and Factors Related to Its Severity.
Fatigue has been reported by many patients undergoing surgery and is associated with a negative prognosis. The factors associated with postoperative fatigue and its evolution during the postoperative period are unclear. Adequate fatigue measurement instruments are necessary to obtain reliable evaluations and to direct effective care to control fatigue in this patient population. This article describes the evolution of postoperative fatigue in patients with cancer as well as related factors. A review of the literature using the CINAHL® and PubMed databases was undertaken. The prevalence of moderate and severe fatigue varies during the postoperative period, with a reduction in the 12 months after surgery. Various factors (e.g., stress, anxiety, depression, pain, changes in sleep patterns) seem to influence the severity of fatigue. More evidence is needed to explore the relationship between immediate postoperative fatigue and the evolution of fatigue during the period following surgical treatment for cancer.


Use of (99m)Tc-labeled liposomes encapsulating blue dye for identification of the sentinel lymph node.
Colloidal radiopharmaceuticals are commonly used in combination with blue dye for localization of the sentinel node. Liposomes are colloidal particles composed of spontaneously forming lipid spheres that can carry a wide variety of diagnostic and therapeutic agents. Conventional liposomes are poorly retained in lymph nodes (<2% of the subcutaneously injected dose). We have previously described a system for increasing the retention of liposomes in the lymph nodes by approximately 7-fold. This system is comprised of subcutaneously injected biotin-coated liposomes, followed by an adjacent injection of avidin. When the avidin moves into the lymphatic vessels, it causes aggregation of the biotin-coated liposomes that are also in the process of migrating through the lymphatic vessels. These aggregated liposomes become entrapped in the next encountered lymph node. In this study, we use this novel lymph node delivery system with liposomes that encapsulate blue dye, resulting in intense blue staining of the sentinel node. These liposomes can also be labeled with (99m)Tc, permitting scintigraphic imaging and radioguided probe localization of the sentinel node. Liposomes coated with biotin and coencapsulating blue dye and glutathione were labeled with (99m)Tc using hexamethylpropyleneamine oxime. Rabbits were subcutaneously administered 0.3 mL (99m)Tc-biotin-liposomes containing blue dye in both hind feet, followed by a subcutaneous injection (0.3 mL) of 5 mg avidin in only one hind foot (experimental). The other hind foot served as a control. Labeling efficiencies (mean +/- SEM) for liposomes encapsulating blue dye were 92.1% +/- 1.9%. Necropsy at 24 h revealed that the popliteal node on the experimental leg receiving the avidin was intensely blue stained compared with virtually no blue coloration of the control node. Tissue counts of these nodes were 12.2 +/- 1.5 percentage injected dose (%ID) in the experimental node compared with 1.2 +/- 0.1 %ID in the control nodes (P<0.0001). Biotin-liposomes encapsulating blue dye can be successfully labeled with (99m)Tc, providing a convenient option for the visualization and radiolocalization of the sentinel node. This biotin-liposome/avidin system may also have potential for the delivery of therapeutic drugs and radiopharmaceuticals to lymph nodes.


Does the patients' expectations on kinesiotape affect the outcomes of patients with a rotator cuff tear? A randomized controlled clinical trial.
To investigate the effect of setting expectations verbally on the effectiveness of kinesiotape application in patients with a rotator cuff tear. Randomized controlled, double-blind study. Department of Physiotherapy and Rehabilitation. Eighty-nine patients with rotator cuff tear. Patients were randomized according to the verbal input given to patients about the effectiveness of kinesiotaping; Group 1 (there is no evidence that kinesiotaping is effective), Group 2 (there is limited evidence that kinesiotaping is effective), and Group 3 (there is evidence that kinesiotaping has an excellent effect). Resting pain, activity pain, and night pain were assessed by visual analog scale. Range of motion was assessed by a universal goniometer. Function was evaluated by the Disabilities of the Arm, Shoulder and Hand Questionnaire and the American Shoulder and Elbow Surgeons Standardized Shoulder Assessment Form before and 24 hours after kinesiotape application. Only resting pain and activity pain were assessed after 30 minutes. There were no statistically significant differences (ANOVA) between any groups at the three assessment points. The intragroup assessment showed that in Group 2, only resting pain after 30 minutes improved (3.2 ± 2.9 to 2.6 ± 2.8; P = 0.02 ). An improvement in resting pain both after 30 minutes and after 24 hours was found in the third group (4.1 ± 2.4 to 2.3 ± 2.3, P = 0.001; 4.1 ± 2.4 to 2.2 ± 2.3, P = 0.001, respectively). Activity pain and night pain were improved in all groups after 24 hours. Setting positive expectations verbally about kinesiotaping might be effective in reducing pain in patients with rotator cuff tear.


Iloprost- and isoproterenol-induced increases in cAMP are regulated by different phosphodiesterases in erythrocytes of both rabbits and humans.
Activation of the G protein G(s) results in increases in cAMP, a necessary step in the pathway for ATP release from rabbit and human erythrocytes. In all cells, the level of cAMP is the product of its synthesis by adenylyl cyclase and its hydrolysis by phosphodiesterases (PDEs). Both iloprost (Ilo), a PGI(2) analog, and isoproterenol (Iso), a beta-agonist, stimulate receptor-mediated increases in cAMP in rabbit and human erythrocytes. However, the specific PDEs associated with each of these signaling pathways in the erythrocyte have not been fully characterized. Previously, we reported that PDE3B is present in rabbit and human erythrocyte membranes and that PDE3 inhibitors potentiate Ilo-induced increases in cAMP. Here we report that inhibitors of either PDE2 or PDE4, erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and rolipram, respectively, potentiate Iso-induced increases in cAMP in rabbit and human erythrocytes. Importantly, these inhibitors had no effect on cAMP increases associated with the incubation of erythrocytes with Ilo. In addition, we establish, for the first time, the presence of PDE2A protein in rabbit and human erythrocyte membranes. Finally, we determined that preincubation of human erythrocytes with EHNA and rolipram together potentiate Iso-induced ATP release, whereas preincubation with cilostazol enhances Ilo-induced release of ATP. These results are consistent with the hypothesis that, in rabbit and human erythrocytes, Ilo-induced increases in cAMP and ATP release are regulated by PDE3, whereas those associated with Iso are regulated by the activities of both PDE2 and PDE4. These studies demonstrate that PDE activity in these cells is localized to specific signaling pathways.


A competing risks prescription refill model of compliance and persistence.
There is evidence to suggest that noncompliant and nonpersistent behaviors have differing risk factors, clinical consequences, and responses to intervention. This has led to calls for these behaviors to be defined and measured separately to characterize medication-taking behavior comprehensively. Current prescription refill models of compliance are, however, unable to appropriately distinguish between noncompliant and nonpersistent behaviors. To address this limitation, a prescription refill model of medication-taking behavior in which noncompliance and nonpersistence are treated as competing risks is presented. The proposed competing risks model of compliance and persistence is compared with a selection of widely applied prescription refill models of compliance and persistence using a common cohort of patients prescribed statin therapy. The competing risks model allows the simultaneous measurement of noncompliance and nonpersistence, the partitioning of their individual contributions to medication-taking behavior, and the estimation of noncompliance risk for patients with varying treatment persistence. The results from this model provide information about the relative and overall contributions of noncompliant and nonpersistent behaviors to medication-taking behavior. The methodology also allows an assessment of the differential influence of various risk factors on these behaviors. The proposed competing risks model differentiates between noncompliant and nonpersistent behaviors using prescription refill data. Results from the model provide insights into the dynamics of noncompliant and nonpersistent behaviors that have not been possible with current prescription refill methodologies.


Red wine extract preserves tight junctions in intestinal epithelial cells under inflammatory conditions: implications for intestinal inflammation.
The altered expression and subcellular distribution of tight junction (TJ) proteins, leading to a dysfunctional intestinal barrier, is a key mechanistic feature of inflammatory bowel disease (IBD). Therefore, increasing the integrity of the intestinal barrier by manipulating the TJ may constitute an innovative and effective therapeutic strategy in IBD. In this context, recent studies showed that dietary polyphenols are able to protect the intestinal TJ barrier integrity. Here, using a cellular model of intestinal inflammation, consisting of cytokine-stimulated HT-29 colon epithelial cells, we show that a polyphenolic extract obtained from Portuguese red wine (RWE) decreased the paracellular permeability across the cell monolayer compared with the control cells, even in the presence of pro-inflammatory cytokines. The beneficial effect of RWE was exerted at three complementary levels: (1) by promoting a significant increase of the mRNA of key barrier-forming TJ proteins, including occludin, claudin-5 and zonnula occludens (ZO)-1 above the levels observed in the control cells; (2) by preventing the decrease in the expression of these proteins under inflammatory conditions and (3) by averting the increase in claudin-2 mRNA, a channel-forming TJ protein induced by pro-inflammatory cytokines. Taken together, these results strongly suggest that polyphenols presented and consumed in red wine as a mixture can reinforce and protect the intestinal barrier against inflammatory stimulus by affecting the TJ protein expression and, thus, without the need for purifying individual compounds, might represent a readily available therapeutic intervention against IBD and intestinal inflammation.


Positron emission tomography in schizophrenia: a new perspective.
PET is an important functional imaging technique that can be used to investigate neurotransmitter receptors and transporters directly by mapping human brain function. PET is increasingly being used greatly to advance our understanding of the neurobiology and pathophysiology of schizophrenia. This review focuses on the use of PET tracers and kinetic modeling in identifying regional brain abnormalities and regions associated with cognitive functioning in schizophrenia. A variety of PET tracers have been used to identify brain abnormalities, including (11)C, (15)O-water, (18)F-fallypride, and L-3,4-dihydroxy-6-(18)F-fluorophenylalanine ((18)F-FDOPA). Some studies have used compartmental modeling to determine tracer binding kinetics. The most consistent findings show a difference in the dopamine content in the prefrontal cortex, anterior cingulate gyrus, and hippocampus between healthy controls and patients with schizophrenia. Studies also show a higher density of D(2) receptors in the striatum and neural brain dysconnectivity. Future investigations integrating clinical, imaging, genetic, and cognitive aspects are warranted to gain a better understanding of the pathophysiology of this disorder.


Mephedrone toxicity in a Scottish emergency department.
To review and improve the evidence base regarding the toxic effects of Mephedrone, a cathinone derivative which was first believed to have been synthesised and propagated in 2007. On the 7th April 2010 the Misuse of Drugs Act 1971 (Amendment) Order 2010 was passed, declaring 4-Methylmethcathinone and other substituted cathinones Class B drugs as of 16 April 2010. This took place despite a perceived lack of evidence as to its potential harms. We undertook a critical review of the available literature to see what evidence existed for the potential effects of mephedrone ingestion. We then conducted our own retrospective consecutive case series of all patients presenting to the Aberdeen Royal Infirmary Emergency Department from the 1st December until the 16th April (the date at which Mephedrone was officially proscripted). The notes were examined for amounts, timings, sources and reasons for ingestion. Symptoms, biometric, ECG and blood analysis data were also recorded along with management and disposal outcomes. 11 articles relating to mephedrone toxicity were identified and reviewed. 89 subjects were found for our case series. This compared to only 27 patients presenting with ingestion of substances (heroin, cocaine, cannabis) at that time illegal. 30 patients stated ingestion of mephedrone in isolation and a further 27 patients stated ingestion of mephedrone with alcohol in addition. A profile of largely psychoactive and cardiovascular toxicity is described with drug naivety perhaps explaining the high rates of bingeing and addiction reported.


Abnormal regenerative responses and impaired axonal outgrowth after nerve crush in TDP-43 transgenic mouse models of amyotrophic lateral sclerosis.
Tar DNA binding protein 43 (TDP-43) mislocalization and aggregation is a hallmark of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar dementia. Moreover, TDP-43 mRNA was found to be upregulated by ∼2.5-fold in the spinal cord of sporadic ALS subjects. Here we have examined the effects of nerve injury in new transgenic mouse models overexpressing by approximately threefold wild-type or mutant (G348C) TDP-43 species. Four weeks after axonal crush of sciatic nerve, TDP-43 transgenic mice remained paralyzed at the injured limb unlike control mice, which had regained most of their normal mobility. In contrast to normal mice, TDP-43 transgenic mice exhibited sustained elevation of TDP-43 cytoplasmic levels in motor neurons after nerve crush, and the relocalization of TDP-43 to the nucleus was delayed by several weeks. After crush, peripherin and ubiquitin levels remained also significantly elevated in TDP-43 transgenic mice compared with control mice. Analysis of the sciatic nerve at 11 d after nerve crush showed that the number of regenerating axons in the distal portion of the lesion was considerably reduced in TDP-43 transgenic mice, especially in TDP-43(G348C) mice, which exhibited a reduction of ∼40%. In addition, markers of neuroinflammation were detected at much higher levels in TDP-43 transgenic mice. These results suggest that a deregulation of TDP-43 expression in ALS is a phenomenon that can affect the regenerative responses to neuronal injury and regrowth potential of axons.


Association between clinical risk factors and progression of chronic kidney disease in children.
Children with chronic kidney disease (CKD) have an increased risk of progression to ESRD. There is a need to identify treatments to slow the progression of CKD, yet there are limited data regarding clinical risk factors that may be suitable targets to slow progression. We performed a retrospective cohort study using the North American Pediatric Renal Trials and Cooperative Studies CKD database. There were 4166 pediatric subjects with CKD stages II to IV. Disease progression was defined as a GFR on follow-up of <15 ml/min per 1.73 m(2) or termination in the registry because of dialysis or transplantation. We used Kaplan-Meier and Cox proportional hazards methods to describe progression rates and determine factors associated with CKD progression. In the univariate analysis, CKD progression was associated with age, gender, race, primary disease, CKD stage, registration year, hematocrit, albumin, corrected calcium, corrected phosphorus, and use of certain medications. Factors that remained significant in the multivariate analysis were age, primary disease, CKD stage, registration year, hypertension, corrected phosphorus, corrected calcium, albumin, hematocrit, and medication proxies for anemia and short stature. There are multiple risk factors associated with disease progression in the pediatric CKD population. Factors that may be amenable to intervention include anemia, hypoalbuminemia, hyperphosphatemia, hypocalcemia, hypertension, and short stature. Because of the retrospective nature of our study, confirmation of our results from ongoing prospective studies is warranted before recommending prospective interventional trials.


cis-Decalin oxidation as a stereochemical probe of in-MOF versus on-MOF catalysis.
Development of catalyst-controlled C-H hydroxylation could provide direct access to valuable synthetic targets, such as primary metabolites. Here, we report a new family of porous materials, comprised of 2-dimensional metalloporphyrin layers and flexible aliphatic linkers, and demonstrate C-H hydroxylation activity. We demonstrate that the stereochemistry of cis-decalin oxidation provides a useful tool for differentiating catalysis in from catalysis on porous materials, which is critical to leveraging the potential of porous materials for catalyst-controlled oxidation chemistry.


Duration of sleep at 3 years of age is associated with fat and fat-free mass at 4 years of age: the Southampton Women's Survey.
Many studies have shown that shorter sleep duration in childhood is associated with higher body mass index (BMI), and have proposed that it is due to an effect of sleep on adiposity. There is little evidence about the association of sleep with fat-free mass. This study examined the association between child's sleep duration at age 3 years and fat and fat-free mass at 4 years of age in a prospective cohort study of 302 boys and 285 girls. Study participants were taking part in the Southampton Women's Survey, a longitudinal study of mothers and children from preconception onwards. Total sleep duration at age 3 years was derived from parental report of night sleep and nap duration. Body composition was assessed by Dual-energy X-ray Absorptiometry (DXA) at 4 years. Mean total sleep duration was 11.5 hours. In linear regression analyses, adjusted for potentially confounding factors (maternal educational attainment, prepregnancy BMI, smoking during pregnancy, child's gestational age at birth, age at DXA, sex, age last breastfed, dietary quality at 3 years, TV watching and hours actively on the move and parental social class), shorter sleep in hours was associated with higher BMI (kg/m(2) ) [β: -0.2340, 95% confidence interval (CI): -0.373 to -0.096], a greater fat mass index (kg) (β: -0.1182, 95% CI: -0.218 to -0.018) and a greater fat-free mass index (kg) (β: -0.100, 95% CI: -0.185 to -0.015). Previous research suggested that the association between shorter sleep and higher body mass index is due to an effect on adiposity. Our findings are novel, suggesting that the relationship between sleep and BMI is also determined by an effect on muscle.


Association between white matter microstructure, executive functions, and processing speed in older adults: the impact of vascular health.
Cerebral white matter damage is not only a commonly reported consequence of healthy aging, but is also associated with cognitive decline and dementia. The aetiology of this damage is unclear; however, individuals with hypertension have a greater burden of white matter signal abnormalities (WMSA) on MR imaging than those without hypertension. It is therefore possible that elevated blood pressure (BP) impacts white matter tissue structure which in turn has a negative impact on cognition. However, little information exists about whether vascular health indexed by BP mediates the relationship between cognition and white matter tissue structure. We used diffusion tensor imaging to examine the impact of vascular health on regional associations between white matter integrity and cognition in healthy older adults spanning the normotensive to moderate-severe hypertensive BP range (43-87 years; N = 128). We examined how white matter structure was associated with performance on tests of two cognitive domains, executive functioning (EF) and processing speed (PS), and how patterns of regional associations were modified by BP and WMSA. Multiple linear regression and structural equation models demonstrated associations between tissue structure, EF and PS in frontal, temporal, parietal, and occipital white matter regions. Radial diffusivity was more prominently associated with performance than axial diffusivity. BP only minimally influenced the relationship between white matter integrity, EF and PS. However, WMSA volume had a major impact on neurocognitive associations. This suggests that, although BP and WMSA are causally related, these differential metrics of vascular health may act via independent pathways to influence brain structure, EF and PS.


Two-year follow-up of amyloid deposition in patients with Alzheimer's disease.
Beta amyloid is one of the major histopathological hallmarks of Alzheimer's disease. We recently reported in vivo imaging of amyloid in 16 Alzheimer patients, using the PET ligand N-methyl[11C]2-(4'-methylaminophenyl)-6-hydroxy-benzothiazole (PIB). In the present study we rescanned these 16 Alzheimer patients after 2.0 +/- 0.5 years and have described the interval change in amyloid deposition and regional cerebral metabolic rate for glucose (rCMRGlc) at follow-up. Sixteen patients with Alzheimer's disease were re-examined by means of PET, using PIB and 2-[18F]fluoro-2-deoxy-d-glucose (FDG) after 2.0 +/- 0.5 years. The patients were all on cholinesterase inhibitor treatment and five also on treatment with the N-methyl-d-aspartate (NMDA) antagonist memantine. In order to estimate the accuracy of the PET PIB measurements, four additional Alzheimer patients underwent repeated examinations with PIB within 20 days (test-retest). Relative PIB retention in cortical regions differed by 3-7% in the test-retest study. No significant difference in PIB retention was observed between baseline and follow-up while a significant (P < 0.01) 20% decrease in rCMRGlc was observed in cortical brain regions. A significant negative correlation between rCMRGlc and PIB retention was observed in the parietal cortex in the Alzheimer patients at follow-up (r = 0.67, P = 0.009). A non-significant decline in Mini-Mental State Examination (MMSE) score from 24.3 +/- 3.7 (mean +/- standard deviation) to 22.7 +/- 6.1 was measured at follow-up. Five of the Alzheimer patients showed a significant decline in MMSE score of >3 (21.4 +/- 3.5 to 15.6 +/- 3.9, P < 0.01) (AD-progressive) while the rest of the patients were cognitively more stable (MMSE score = 25.6 +/- 3.1 to 25.9 +/- 3.7) (AD-stable) compared with baseline. A positive correlation (P = 0.001) was observed in the parietal cortex between Rey Auditory Verbal Learning (RAVL) test score and rCMRGlc at follow-up while a negative correlation (P = 0.018) was observed between RAVL test and PIB retention in the parietal at follow-up. Relatively stable PIB retention after 2 years of follow-up in patients with mild Alzheimer's disease suggests that amyloid deposition in the brain reaches a plateau by the early clinical stages of Alzheimer's disease and therefore may precede a decline in rCMRGlc and cognition. It appears that anti-amyloid therapies will need to induce a significant decrease in amyloid load in order for PIB PET images to detect a drug effect in Alzheimer patients. FDG imaging may be able to detect a stabilization of cerebral metabolism caused by therapy administered to patients with a clinical diagnosis of Alzheimer's disease.


Prognostic significance of angiogenesis evaluated by CD105 expression compared to CD31 in 905 breast carcinomas: correlation with long-term patient outcome.
Our purpose was to determine the respective prognostic significance of CD105 and CD31 immunoexpression in node negative patients with breast carcinoma, since angiogenesis induces blood borne metastases and death in carcinomas. CD105 (endoglin) has been reported as expressed by activated endothelial cells and consequently should better reflect neoangiogenesis in malignant tumors. Comparison of CD31 and CD105 immunocytochemical expression was undertaken in a series of 905 breast carcinomas. Results were compared to patients' long-term (median = 11.3 years) outcome. Univariate (Kaplan-Meier) analysis showed that the number of CD105+ microvessels (cut-off 15 vessels) correlated significantly with poor overall survival (p=0.001). This correlation was less significant in node negative patients (p=0.035). The number of CD31+ microvessels (cut-off 25 vessels) similarly correlated with poor survival (p=0.032) but not in the subgroup of node negative patients. Marked CD105 expression also correlated with a high risk for metastasis in all patients (p=0.0002) and in the subset of node negative patients (p=0.001). Similarly metastasis risk in node negative patients correlated with marked CD31 immunocytochemical expression (p=0.02). Multivariate analysis (Cox model) identified CD105, but not CD31 immunoexpression, as an independent prognostic indicator. Our results suggest that: i) in breast carcinomas, immunoselection of microvessels containing activated CD105 labelled endothelial cells is endowed with a stronger prognostic significance, as compared to CD31 vessels labelling; ii) the CD105 immunoexpression may be considered as a potential tool for selecting node negative patients with a poorer outcome and higher metastasis risk; iii) in these patients specific antiangiogenic therapy targeted by anti-CD105 conjugates can be further developed.


LncIHAT Is Induced by Hypoxia-Inducible Factor 1 and Promotes Breast Cancer Progression.
Hypoxia induces thousands of mRNAs and miRNAs to mediate tumor malignancy. However, hypoxia-induced long noncoding RNA (lncRNA) transcriptome and their role in triple-negative breast cancer (TNBC) have not been defined. Here we identified hypoxia-induced lncRNA transcriptome in two human TNBC cell lines by whole transcriptome sequencing. AC093818.1 was one of 26 validated lncRNAs and abundantly expressed in TNBC in vitro and in vivo. 5'- and 3'-rapid amplification of cDNA ends assays revealed that the isoform 2 was a dominant AC093818.1 transcript in TNBC cells and thus referred to as lncIHAT (lncRNA induced by hypoxia and abundant in TNBC). Hypoxia-inducible factor 1 (HIF1) but not HIF2 bound to the hypoxia response element at the promoter of lncIHAT to activate its transcription in hypoxic TNBC cells. LncIHAT promoted TNBC cell survival in vitro and tumor growth and lung metastasis in mice. Mechanistically, lncIHAT was required for the expression of its proximal neighboring oncogenic genes PDK1 and ITGA6 in TNBC cells and tumors. Reexpression of PDK1 and ITGA6 rescued survival and growth of lncIHAT knockdown TNBC cells in vitro. Collectively, these findings uncovered lncIHAT as a new hypoxia-induced oncogenic cis-acting lncRNA in TNBC. IMPLICATIONS: This study systematically identified hypoxia-induced lncRNA transcriptome in TNBC and sheds light on multiple layers of regulatory mechanisms of gene expression under hypoxia.


[How to manage chronic kidney disease before dialysis].
Chronic renal failure is usually a silent disease until its late stage, especially in elderly people. Screening for such disease is particularly useful in hypertensive diabetic patients above 50 years. The causes are indeed often vascular or metabolic (directly or not directly linked to diabetes mellitus). Other less frequent causes are yet possible. The search for the right diagnosis of renal insufficiency is always requested to apply the appropriate treatment, combined with medical measures for secondary and tertiary prevention. This review will give general advices to avoid the development of renal disease (stages 3 and 4) or its progression, and also insist on the potential nephrotoxic effects of some drugs.


Brain Transcriptomic Analysis of Hereditary Cerebral Hemorrhage With Amyloidosis-Dutch Type.
Hereditary cerebral hemorrhage with amyloidosis-Dutch type (HCHWA-D) is an early onset hereditary form of cerebral amyloid angiopathy (CAA) caused by a point mutation resulting in an amino acid change (NP_000475.1:p.Glu693Gln) in the amyloid precursor protein (APP). Post-mortem frontal and occipital cortical brain tissue from nine patients and nine age-related controls was used for RNA sequencing to identify biological pathways affected in HCHWA-D. Although previous studies indicated that pathology is more severe in the occipital lobe in HCHWA-D compared to the frontal lobe, the current study showed similar changes in gene expression in frontal and occipital cortex and the two brain regions were pooled for further analysis. Significantly altered pathways were analyzed using gene set enrichment analysis (GSEA) on 2036 significantly differentially expressed genes. Main pathways over-represented by down-regulated genes were related to cellular aerobic respiration (including ATP synthesis and carbon metabolism) indicating a mitochondrial dysfunction. Principal up-regulated pathways were extracellular matrix (ECM)-receptor interaction and ECM proteoglycans in relation with an increase in the transforming growth factor beta (TGFβ) signaling pathway. Comparison with the publicly available dataset from pre-symptomatic APP-E693Q transgenic mice identified overlap for the ECM-receptor interaction pathway, indicating that ECM modification is an early disease specific pathomechanism.


Exposure to glyphosate-based herbicides and risk for non-Hodgkin lymphoma: A meta-analysis and supporting evidence.
Glyphosate is the most widely used broad-spectrum systemic herbicide in the world. Recent evaluations of the carcinogenic potential of glyphosate-based herbicides (GBHs) by various regional, national, and international agencies have engendered controversy. We investigated whether there was an association between high cumulative exposures to GBHs and increased risk of non-Hodgkin lymphoma (NHL) in humans. We conducted a new meta-analysis that includes the most recent update of the Agricultural Health Study (AHS) cohort published in 2018 along with five case-control studies. Using the highest exposure groups when available in each study, we report the overall meta-relative risk (meta-RR) of NHL in GBH-exposed individuals was increased by 41% (meta-RR = 1.41, 95% confidence interval, CI: 1.13-1.75). For comparison, we also performed a secondary meta-analysis using high-exposure groups with the earlier AHS (2005), and we calculated a meta-RR for NHL of 1.45 (95% CI: 1.11-1.91), which was higher than the meta-RRs reported previously. Multiple sensitivity tests conducted to assess the validity of our findings did not reveal meaningful differences from our primary estimated meta-RR. To contextualize our findings of an increased NHL risk in individuals with high GBH exposure, we reviewed publicly available animal and mechanistic studies related to lymphoma. We documented further support from studies of malignant lymphoma incidence in mice treated with pure glyphosate, as well as potential links between glyphosate / GBH exposure and immunosuppression, endocrine disruption, and genetic alterations that are commonly associated with NHL or lymphomagenesis. Overall, in accordance with findings from experimental animal and mechanistic studies, our current meta-analysis of human epidemiological studies suggests a compelling link between exposures to GBHs and increased risk for NHL.


Randomized perturbed posturography in abstinent chronic alcoholics.
Equilibrium is maintained by visual, proprioceptive and vestibular afferent influx, central coordination and motor efferents. Eleven male abstinent chronic alcoholic volunteers aged 44-65 years (mean 57 years) were investigated by randomized perturbed posturography. They had all been long time heavy drinkers. Ocular smooth pursuit and visual suppression of the vestibulo-ocular reflex showed abnormalities in a previous study, but they had no signs or symptoms of polyneuropathy. The current study demonstrated larger (but generally not significantly so) sway areas, and in all tests on the randomly moving support surface the sway velocities were significantly larger than in the controls. It is concluded that randomized perturbed posturography is valuable in dysequilibrium assessment in chronic alcoholics.


Hybridization kinetics is different inside cells.
It is generally expected that the kinetics of reactions inside living cells differs from the situation in bulk solutions. Macromolecular crowding and specific binding interactions could change the diffusion properties and the availability of free molecules. Their impact on reaction kinetics in the relevant context of living cells is still elusive, mainly because the difficulty of capturing fast kinetics in vivo. This article shows spatially resolved measurements of DNA hybridization kinetics in single living cells. HeLa cells were transfected with a FRET-labeled dsDNA probe by lipofection. We characterized the hybridization reaction kinetics with a kinetic range of 10 micros to 1 s by a combination of laser-driven temperature oscillations and stroboscopic fluorescence imaging. The time constant of the hybridization depended on DNA concentration within individual cells and between cells. A quantitative analysis of the concentration dependence revealed several-fold accelerated kinetics as compared with free solution for a 16-bp probe and decelerated kinetics for a 12-bp probe. We did not find significant effects of crowding agents on the hybridization kinetics in vitro. Our results suggest that the reaction rates in vivo are specifically modulated by binding interactions for the two probes, possibly triggered by their different lengths. In general, the presented imaging modality of temperature oscillation optical lock-in microscopy allows to probe biomolecular interactions in different cell compartments in living cells for systems biology.


General practice out-of-hours service, variations in use and equality in access to a doctor: a cross-sectional study.
There is increasing dissatisfaction among general practitioners (GPs) with their 24 hour commitment despite the demand for GP services outside ordinary working hours. The creation of out-of-hours co-operatives has been welcomed by participating GPs and their families and patient satisfaction is generally high. However, there have been concerns about the lack of patient consultation in their development and on the quality of care provided. To examine geographical variation in the rates of out-of-hours calls and to see if there is any systematic variation in how the co-operative that covers the area responds to calls. A secondary data analysis of 110,357 calls received by the co-operative during the second year of its operation. A co-operative with four centres providing out-of-hours care for one mostly rural Health Board in Northern Ireland. Deprivation score and mortality and long-term illness ratios provided indicators of need at an area level. Proximity to the four co-operative centres was measured as the distance in kilometres and estimated travel time (in minutes) along the road network. Out-of-hours call ratios were proportional to the area deprivation score and proximity to the co-operative centres, though not to area indicators of ill health. Older patients were more likely to be seen by the GP and females over the age of 15 years were more likely to receive telephone advice only (adjusted odds ratio [OR] = 0.785, 95% confidence intervals [CI] 0.756-0.816, P < 0.001). Each kilometre from the centres reduced the likelihood of seeing the GP (OR = 0.978, 95% CI = 0.976-0.979, P < 0.001). Even after controlling for potential confounders a large difference remained in how centres responded to calls. It is not certain whether these inequalities in services delivery represent inequity as the apposite level or type of response cannot be determined until more is known about the appropriateness of the demand for out-of-hours medical care.


Heritability of language laterality assessed by functional transcranial Doppler ultrasound: a twin study.
Background: Prior studies have estimated heritability of around 0.25 for the trait of handedness, with studies of structural brain asymmetry giving estimates in a similar or lower range. Little is known about heritability of functional language lateralization. This report describes heritability estimates using functional language laterality and handedness phenotypes in a twin sample previously reported by Wilson and Bishop (2018). Methods: The total sample consisted of 194 twin pairs (49% monozygotic) aged from 6 to 11 years. A language laterality index was obtained for 141 twin pairs, who completed a protocol where relative blood flow through left and right middle cerebral arteries was measured using functional transcranial Doppler ultrasound (fTCD) while the child described animation sequences. Handedness data was available from the Edinburgh Handedness Inventory (EHI) and Quantification of Hand Preference (QHP) for all 194 pairs. Heritability was assessed using conventional structural equation modeling, assuming no effect of shared environment (AE model). Results: For the two handedness measures, heritability estimates were consistent with prior research: 0.23 and 0.22 respectively for the EHI and QHP. For the language laterality index, however, the twin-cotwin correlations were close to zero for both MZ and DZ twins, and the heritability estimate was zero. Conclusions: A single study cannot rule out a genetic effect on language lateralisation. It is possible that the low twin-cotwin correlations were affected by noisy data: although the split-half reliability of the fTCD-based laterality index was high (0.85), we did not have information on test-retest reliability in children, which is likely to be lower. We cannot reject the hypothesis that there is low but nonzero heritability for this trait, but our data suggest that individual variation in language lateralisation is predominantly due to stochastic variation in neurodevelopment.


Caprylic triglyceride as a novel therapeutic approach to effectively improve the performance and attenuate the symptoms due to the motor neuron loss in ALS disease.
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder of motor neurons causing progressive muscle weakness, paralysis, and finally death. ALS patients suffer from asthenia and their progressive weakness negatively impacts quality of life, limiting their daily activities. They have impaired energy balance linked to lower activity of mitochondrial electron transport chain enzymes in ALS spinal cord, suggesting that improving mitochondrial function may present a therapeutic approach for ALS. When fed a ketogenic diet, the G93A ALS mouse shows a significant increase in serum ketones as well as a significantly slower progression of weakness and lower mortality rate. In this study, we treated SOD1-G93A mice with caprylic triglyceride, a medium chain triglyceride that is metabolized into ketone bodies and can serve as an alternate energy substrate for neuronal metabolism. Treatment with caprylic triglyceride attenuated progression of weakness and protected spinal cord motor neuron loss in SOD1-G93A transgenic animals, significantly improving their performance even though there was no significant benefit regarding the survival of the ALS transgenic animals. We found that caprylic triglyceride significantly promoted the mitochondrial oxygen consumption rate in vivo. Our results demonstrated that caprylic triglyceride alleviates ALS-type motor impairment through restoration of energy metabolism in SOD1-G93A ALS mice, especially during the overt stage of the disease. These data indicate the feasibility of using caprylic acid as an easily administered treatment with a high impact on the quality of life of ALS patients.


Use of CA-125 to assess response to new agents in ovarian cancer trials.
The purpose of this article is to demonstrate how CA-125 could be used in clinical trials to ascertain the efficacy of new drugs for ovarian cancer. Studies that have investigated the use of CA-125 in assessing response and progression of ovarian cancer are reviewed. A precise CA-125 response definition that requires either a 50% or 75% decrease in CA-125 levels has been shown in trials of both initial chemotherapy and in phase II trials to predict accurately the response in comparison with standard response criteria. A simpler response definition that is based on just a 50% decrease in CA-125 levels has been proposed by the Gynaecological Cancer Intergroup (GCIG) but requires further validation. Definitions for progression have also been proposed by the GCIG on the basis of a confirmed doubling of CA-125 levels from either the upper limit of normal or the nadir CA-125 level. These CA-125 definitions for progression falsely predict progression in fewer than 2% of patients and can be used to define the date of progression. Precise definitions for response and progression according to CA-125 should be incorporated into ovarian cancer clinical trial protocols. These definitions already have been shown to be valuable in assessing efficacy of new agents but require further prospective evaluation.


The Role of Endometrial Selectins and Their Ligands on Bovine Conceptus Attachment to the Uterine Epithelium During Peri-Implantation Period.
A successful pregnancy depends on the blastocyst's implantation to the maternal endometrium; however, the initial interaction between blastocyst and uterine epithelium has not been well characterized. The objectives of this study were to determine if selectins and their ligands were expressed in the bovine conceptus and/or uterus during the periattachment period and to study whether selectins were associated with conceptus attachment to the uterine epithelium. Through the RNA-sequence analysis of bovine conceptuses on Days 17, 20, and 22 (Day 0 = day of estrus), only the SELL ligand, podocalyxin (PODXL), and P-selectin (SELP) ligand, SELPLG, were found. Quantitative PCR analysis confirmed the presence of PODXL and SELPLG in these conceptuses and revealed that SELL, mRNA and protein, detected in the uterine epithelium but not in conceptuses increased during the periattachment period. In the cultured endometrial epithelial cells (EECs), SELL transcript was up-regulated when uterine flushings from Day 20 pregnant animals were placed onto these cells. SELL was also up-regulated when cultured EECs were treated with progesterone, EGF, or bFGF, but not with IFNT. In the coculture system with EECs and bovine trophoblast CT-1 cells, SELL expression in EECs was effectively reduced by its small interfering RNA; however, IFNT, a marker for CT-1 cell attachment to EECs, was not reduced, nor was a transcription factor of IFNT, CDX2. These observations suggest that the conceptus could attach to the uterine epithelium through the use of endometrial SELL and embryonic selectin ligands, possibly initiating the conceptus attachment process in the bovine species.


The critical role of metabolic pathways in aging.
Aging is characterized by a deterioration in the maintenance of homeostatic processes over time, leading to functional decline and increased risk for disease and death. The aging process is characterized metabolically by insulin resistance, changes in body composition, and physiological declines in growth hormone (GH), insulin-like growth factor-1 (IGF-1), and sex steroids. Some interventions designed to address features of aging, such as caloric restriction or visceral fat depletion, have succeeded in improving insulin action and life span in rodents. Meanwhile, pharmacologic interventions and hormonal perturbations have increased the life span of several mammalian species without necessarily addressing features of age-related metabolic decline. These interventions include inhibition of the mammalian target of rapamycin and lifetime deficiency in GH/IGF-1 signaling. However, strategies to treat aging in humans, such as hormone replacement, have mostly failed to achieve their desired response. We will briefly discuss recent advances in our understanding of the complex role of metabolic pathways in the aging process and highlight important paradoxes that have emerged from these discoveries. Although life span has been the major outcome of interest in the laboratory, a special focus is made in this study on healthspan, as improved quality of life is the goal when translated to humans.


Surfactant protein-D, a mediator of innate lung immunity, alters the products of nitric oxide metabolism.
Surfactant protein (SP)-D, a 43-kD multifunctional collagen-like lectin, is synthesized and secreted by the airway epithelium. SP-D knockout (SP-D [-/-]) mice exhibit an increase in the number and size of airway macrophages, peribronchiolar inflammation, increases in metalloproteinase activity, and development of emphysema. Nitric oxide (NO) is involved in a variety of signaling processes, and because altered NO metabolism has been observed in inflammation, we hypothesized that alterations in its metabolism would underlie the proinflammatory state observed in SP-D deficiency. Examination of the bronchial alveolar lavage (BAL) from SP-D (-/-) mice reveals a significant increase in protein and phospholipid content and total cell count. NO production and inducible NO synthase expression were increased in the BAL; however, there was a decline in S-nitrosothiol (SNO) content in the BAL and a loss of SNO immunoreactivity within the tissue. This decline in SNO was accompanied by an increase in nitrotyrosine staining. We conclude that inflammation that occurs in SP-D deficiency results in an increase in NO production and a shift in the chemistry and targets of NO. We speculate that the proinflammatory response due to SP-D deficiency results, in part, from a disruption of NO-mediated signaling within the innate immune system.


Epidermal tendon cells require Broad Complex function for correct attachment of the indirect flight muscles in Drosophila melanogaster.
Drosophila Broad Complex, a primary response gene in the ecdysone cascade, encodes a family of zinc-finger transcription factors essential for metamorphosis. Broad Complex mutations of the rbp complementation group disrupt attachment of the dorsoventral indirect flight muscles during pupal development. We previously demonstrated that isoform BRC-Z1 mediates the muscle attachment function of rbp(+) and is expressed in both developing muscle fibers and their epidermal attachment sites. We now report two complementary studies to determine the cellular site and mode of action of rbp(+) during maturation of the myotendinous junctions of dorsoventral indirect flight muscles. First, genetic mosaics, produced using the paternal loss method, revealed that the muscle attachment phenotype is determined primarily by the genotype of the dorsal epidermis, with the muscle fiber and the ventral epidermis exerting little or no influence. When the dorsal epidermis was mutant, the vast majority of muscles detached or chose ectopic attachment sites, regardless of the muscle genotype. Conversely, wild-type dorsal epidermis could support attachment of mutant muscles. Second, ultrastructural analysis corroborated and extended these results, revealing defective and delayed differentiation of rbp mutant epidermal tendon cells in the dorsal attachment sites. Tendon cell processes, the stress-bearing links between the epidermis and muscle, were reduced in number and showed delayed appearance of microtubule bundles. In contrast, mutant muscle and ventral epidermis resembled the wild type. In conclusion, BRC-Z1 acts in the dorsal epidermis to ensure differentiation of the myotendinous junction. By analogy with the cell-cell interaction essential for embryonic muscle attachment, we propose that BRC-Z1 regulates one or more components of the epidermal response to a signal from the developing muscle.


Topical Delivery of Anti-VEGF Drugs to the Ocular Posterior Segment Using Cell-Penetrating Peptides.
To evaluate the efficacy of anti-VEGF agents for treating choroidal neovascularization (CNV) when delivered topically using novel cell-penetrating peptides (CPPs) compared with delivery by intravitreal (ivit) injection. CPP toxicity was investigated in cell cultures. Ivit concentrations of ranibizumab and bevacizumab after topical administration were measured using ELISA. The biological efficacy of topical anti-VEGF + CPP complexes was compared with ivit anti-VEGF injections using an established model of CNV. CPPs were nontoxic in vitro. In vivo, after topical eye drop delivery, CPPs were present in the rat anterior chamber within 6 minutes. A single application of CPP + bevacizumab eye drop delivered clinically relevant concentrations of bevacizumab to the posterior chamber of the rat eye in vivo. Similarly, clinically relevant levels of CPP + ranibizumab and CPP + bevacizumab were detected in the porcine vitreous and retina ex vivo. In an established model of CNV, mice treated with either a single ivit injection of anti-VEGF, twice daily CPP + anti-VEGF eye drops or daily dexamethasone gavage for 10 days all had significantly reduced areas of CNV when compared with lasered eyes without treatment. CPPs are nontoxic to ocular cells and can be used to deliver therapeutically relevant doses of ranibizumab and bevacizumab by eye drop to the posterior segment of mouse, rat, and pig eyes. The CPP + anti-VEGF drug complexes were cleared from the retina within 24 hours, suggesting a daily eye drop dosing regimen. Daily, topically delivered anti-VEGF with CPP was as efficacious as a single ivit injection of anti-VEGF in reducing areas of CNV in vivo.


Psychiatric Aeromedical Evacuations of Deployed Active Duty U.S. Military Personnel During Operations Enduring Freedom, Iraqi Freedom, and New Dawn.
The primary objective of this study was to describe the demographic, clinical, and attrition characteristics of active duty U.S. military service members who were aeromedically evacuated from Iraq and Afghanistan theaters with a psychiatric condition as the primary diagnosis. The study links the U.S. Transportation Command Regulating and Command and Control Evacuation System (TRAC2ES) data with the Defense Manpower Data Center (DMDC) to conduct an examination of the long-term occupational impact of psychiatric aeromedical evacuations on military separations and discharges. Retrospective analyses were conducted on the demographic, clinical, and attrition information of active duty service members (N = 7,023) who received a psychiatric aeromedical evacuation from Iraq or Afghanistan between 2001 and 2013 using TRAC2ES data. Additionally, TRAC2ES database was compared with DMDC data to analyze personal and service demographics, aeromedical evacuation information, and reasons for military separation with the entire 2013 active duty force. Chi-square tests of independence and standardized residuals were used to identify cells with observed frequencies or proportions significantly different than expected by chance. Additionally, OR were calculated to provide context about the nature of any significant relationships. Compared with the active duty comparison sample, those with a psychiatric aeromedical evacuation tended to be younger, female, white, divorced or widowed, and less educated. They were also more likely to be junior enlisted service members in the Army serving in a Combat Arms military occupational specialty. The primary psychiatric conditions related to the aeromedical evacuation were depressive disorders (25%), adjustment disorders (18%), post-traumatic stress disorder (9%), bipolar disorders (6%), and anxiety disorders (6%). Approximately, 3% were evacuated for suicidal ideation and associated behaviors. Individuals who received a psychiatric aeromedical evacuation were almost four times as likely (53%) to have been subsequently separated from active duty at the time of the data analysis compared with other active duty service members (14%). The current study also found that peaks in the number of aeromedical evacuations coincided with significant combat operational events. These peaks almost always preceded or followed a significant operational event. An unexpected finding of the present study was that movement classification code was not predictive of subsequent reasons for separation from the military. Thus, the degree of clinical supervision and restraint of a service member during psychiatric aeromedical evacuation from deployment proved to be unrelated to subsequent service outcome. Psychiatric conditions are one of the leading reasons for the aeromedical evacuation of active duty military personnel from the military combat theater. For many active duty military personnel, a psychiatric aeromedical evacuation from a combat theater is the start of a military career-ending event that results in separation from active duty. This finding has important clinical and operational implications for the evaluation and treatment of psychiatric conditions during military deployments. Whenever possible, deployed military behavioral health providers should attempt to treat psychiatric patients in theater to help them remain in theater to complete their operational deployments. Improved understanding of the factors related to psychiatric aeromedical evacuations will provide important clinical and policy implications for future conflicts.


Conformational plasticity in the selectivity filter of the TRPV2 ion channel.
Transient receptor potential vanilloid (TRPV) channels are activated by ligands and heat and are involved in various physiological processes. In contrast to the architecturally related voltage-gated cation channels, TRPV1 and TRPV2 subtypes possess another activation gate at the selectivity filter that can open widely enough to permeate large organic cations. Despite recent structural advances, the mechanism of selectivity filter gating and permeation for both metal ions and large molecules by TRPV1 or TRPV2 is not well known. Here, we determined two crystal structures of rabbit TRPV2 in its Ca2+-bound and resiniferatoxin (RTx)- and Ca2+-bound forms, to 3.9 Å and 3.1 Å, respectively. Notably, our structures show that RTx binding leads to two-fold symmetric opening of the selectivity filter of TRPV2 that is wide enough for large organic cation permeation. Combined with functional characterizations, our studies reveal a structural basis for permeation of Ca2+ and large organic cations in TRPV2.


Use of secondary structural information and C alpha-C alpha distance restraints to model protein structures with MODELLER.
Protein secondary structure predictions and amino acid long range contact map predictions from primary sequence of proteins have been explored to aid in modelling protein tertiary structures. In order to evaluate the usefulness of secondary structure and 3D-residue contact prediction methods to model protein structures we have used the known Q3 (alpha-helix,beta-strands and irregular turns/loops) secondary structure information, along with residue-residue contact information as restraints for MODELLER. We present here results of our modelling studies on 30 best resolved single domain protein structures of varied lengths. The results shows that it is very difficult to obtain useful models even with 100% accurate secondary structure predictions and accurate residue contact predictions for up to 30% of residues in a sequence. The best models that we obtained for proteins of lengths 37, 70, 118, 136 and 193 amino acid residues are of RMSDs 4.17, 5.27, 9.12, 7.89 and 9.69,respectively. The results show that one can obtain better models for the proteins which have high percent of alpha-helix content. This analysis further shows that MODELLER restrain optimization program can be useful only if we have truly homologous structure(s) as a template where it derives numerous restraints, almost identical to the templates used. This analysis also clearly indicates that even if we satisfy several true residue-residue contact distances, up to 30%of their sequence length with fully known secondary structural information, we end up predicting model structures much distant from their corresponding native structures.


Safety and efficacy of the Gunther Tulip retrievable vena cava filter: midterm outcomes.
To evaluate of the medium-term integrity, efficacy, and complication rate associated with the Gunther Tulip vena cava filter. A retrospective study was performed of 369 consecutive patients who had infrarenal Gunther Tulip inferior vena cava filters placed over a 5-year period. The mean patient age was 61.8 years, and 59% were men. Venous thromboembolic disease and a contraindication to or complication of anticoagulation were the indications for filter placement in 86% of patients; 14% were placed for prophylaxis in patients with a mean of 2.3 risk factors. Follow-up was obtained by review of medical and radiologic records. Mean clinical follow-up was 780 days. New or recurrent pulmonary embolus occurred in 12 patients (3.3%). New or recurrent deep-vein thrombosis occurred in 53 patients (14.4%). There were no symptomatic fractures, migrations, or caval perforations. Imaging follow-up in 287 patients (77.8%) at a mean of 731 days revealed a single (0.3%) asymptomatic fracture, migration greater than 2 cm in 36 patients (12.5%), and no case of embolization. Of 122 patients with CT scans, asymptomatic perforations were identified in 53 patients (43.4%) at a mean 757 days. The Gunther Tulip filter was safe and effective at 2-year follow-up. Complication rates were similar to those reported for permanent inferior vena cava filters.


Desialylation of lysosomal membrane glycoproteins by Trypanosoma cruzi: a role for the surface neuraminidase in facilitating parasite entry into the host cell cytoplasm.
Trypanosoma cruzi enters host cells via formation of an acidic vacuole which is subsequently disrupted, allowing the parasite access to the cytoplasm. We show that in an acid environment, release of the parasite surface neuraminidase is enhanced, and this release is likely mediated by a phosphatidylinositol-specific phospholipase C (PIPLC), since antibodies to a carbohydrate epitope (CRD) revealed in glycosylphosphatidylinositol (GPI)-anchored proteins after PIPLC cleavage remove the great majority of the soluble neuraminidase activity from culture supernatants. The neuraminidase is active at acidic pH, and is capable of desialylating known vacuolar constituents, i.e., lysosomal membrane glycoproteins. Parasite escape into the cytoplasm is significantly facilitated in terminal sialylation-defective mutant Lec 2 cells, and enzymatically desialylated membranes are more susceptible to lysis by a parasite hemolysin previously implicated in vacuole membrane rupture. These findings provide evidence that terminal sialylation on carbohydrate moieties contributes to maintaining lysosomal membrane integrity, and indicate a role for a protozoan-derived neuraminidase in facilitating parasite entry into host cells. These observations raise the possibility that other microbial neuraminidases may serve a similar function in acidic intracellular compartments.


Measuring the mutual diffusion coefficient for dodecyl acrylate in low molecular weight poly(dodecyl acrylate) with laser line deflection (Wiener's Method) and the fluorescence of pyrene.
Diffusion of small molecules into glassy polymers is quite complicated and almost always non-Fickian. Little work has been done with the diffusion of low molecular weight polymers that are liquids at room temperature (such as poly(dodecyl acrylate)) into their miscible monomers. We have studied three molecular weights under 20 000 to determine if poly(dodecyl acrylate) diffusion into dodecyl acrylate could be treated with Fick's law and if so to determine the values of the diffusion coefficients. We compare two methods for measuring the diffusion of dodecyl acrylate into poly(dodecyl acrylate): We used laser line deflection (Wiener's method) and improved upon the method from published reports. We also used the dependence of pyrene's fluorescence on the viscosity to measure the concentration distribution, and thus to extract the diffusion coefficient. After an initial relaxation period, diffusion in all cases followed Fick's law with a single concentration-independent diffusion coefficient. Comparison of the diffusion coefficients obtained by both methods yielded the same order of magnitude for the diffusion coefficients (10(-7) cm2/s) and showed the same trend in the dependence on the average molecular weight of the polymer (a decrease in the diffusion coefficient with an increase in the molecular weight).


Endogenous estrogen and postmenopausal breast cancer: a quantitative review.
This paper systematically reviews the results from epidemiologic studies investigating the hypothesis that breast cancer risk in postmenopausal women increases with increasing concentrations of estradiol in blood and with increasing urinary estrogen excretion rates. Data from 29 epidemiologic studies of endogenous hormones and postmenopausal breast cancer were used. The ratio of the average estrogen concentration in the women with breast cancer to that in the women without breast cancer (and its 95 percent confidence interval [CI]) was calculated for each study, and the results were summarized by calculating weighted averages of the log ratios. In six prospective studies of serum estradiol concentration, 329 women who subsequently developed breast cancer had, overall, a 15 percent (CI = 6-24 percent, P = 0.0003) higher mean concentration of estradiol in their blood than the 1,105 women who remained free of cancer. The results of these prospective studies did not differ significantly from each other (chi2 for heterogeneity = 8.7; degrees of freedom = 5; P > 0.1). Similar differences in mean estrogen levels were seen in the case-control studies which reported either estradiol concentrations in the blood or urinary estrogen excretion. However, the case-control studies showed significant heterogeneity among their results. The data from the prospective studies strongly suggest that breast cancer risk in postmenopausal women is associated with relatively high concentrations of endogenous estradiol.


Use and environmental occurrence of antibiotics in freestall dairy farms with manured forage fields.
Environmental releases of antibiotics from concentrated animal feeding operations (CAFOs) are of increasing regulatory concern. This study investigates the use and occurrence of antibiotics in dairy CAFOs and their potential transport into first-encountered groundwater. On two dairies we conducted four seasonal sampling campaigns, each across 13 animal production and waste management systems and associated environmental pathways: application to animals, excretion to surfaces, manure collection systems, soils, and shallow groundwater. Concentrations of antibiotics were determined using on line solid phase extraction (OLSPE) and liquid chromatography-tandem mass spectrometry (LC/MS/MS) with electrospray ionization (ESI) for water samples, and accelerated solvent extraction (ASE) LC/MS/MS with ESI for solid samples. A variety of antibiotics were applied at both farms leading to antibiotics excretion of several hundred grams per farm per day. Sulfonamides, tetracyclines, and their epimers/isomers, and lincomycin were most frequently detected. Yet, despite decades of use, antibiotic occurrence appeared constrained to within farm boundaries. The most frequent antibiotic detections were associated with lagoons, hospital pens, and calf hutches. When detected below ground, tetracyclines were mainly found in soils, whereas sulfonamides were found in shallow groundwater reflecting key differences in their physicochemical properties. In manure lagoons, 10 compounds were detected including tetracyclines and trimethoprim. Of these 10, sulfadimethoxine, sulfamethazine, and lincomycin were found in shallow groundwater directly downgradient from the lagoons. Antibiotics were sporadically detected in field surface samples on fields with manure applications, but not in underlying sandy soils. Sulfadimethoxine and sulfamethazine were detected in shallow groundwater near field flood irrigation gates, but at highly attenuated levels.


Physics for infants: characterizing the origins of knowledge about objects, substances, and number.
Adults possess a great deal of knowledge about how objects behave and interact in our every day environment, yet several puzzles remain unsolved regarding how we manage this ubiquitous skill. The notion of intuitive physics has been a central focus of research on cognitive development in infancy. This article focuses on the origins of knowledge about objects, substances, and number concepts in infancy. The article reviews common themes of solidity, continuity, cohesion, and property changes as they have been studied with regard to infants' knowledge about objects and more recently with regard to infants' knowledge about substances. In addition, we review how object and substance knowledge interfaces with number knowledge systems. The evidence supports the view that certain core principles about these domains are present as early as we can test for them and the nature of the underlying representation is best characterized as primitive initial concepts that are elaborated and refined through learning and experience. WIREs Cogn Sci 2012, 3:19-27. doi: 10.1002/wcs.157 For further resources related to this article, please visit the WIREs website.


Alcohol and Drug Use Before and During the First Year After Traumatic Brain Injury.
To compare individuals with mild and moderate/severe traumatic brain injury (TBI) on alcohol and drug use and substance use disorders before and in the first year post-TBI; to explore sociodemographic and injury-related variables associated with substance use disorders. A total of 225 adults hospitalized in a level I trauma center after TBI. Observational cohort study with retrospective (pre-TBI) and prospective (4, 8, and 12 months post-TBI) assessments. Mini International Neuropsychiatric Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). The percentage of participants using alcohol or drug declined shortly after the injury (4 months) but increased closer to preinjury levels by the end of the first year. Post-TBI alcohol use was higher after mild than moderate/severe TBI, but drug use was similar. About 11% of participants met criteria for a substance use disorder in the first year after TBI. Younger age, not being in a relationship, and suspected substance intoxication at the time of TBI were associated with the presence of a post-TBI substance use disorder. Individuals with milder injuries return to alcohol use earlier than those with more severe injuries. Given that substance use may alter recovery, preventive recommendations and systematic follow-ups are warranted regardless of injury severity and access to rehabilitation.


Large-scale genome sampling reveals unique immunity and metabolic adaptations in bats.
Comprising more than 1,400 species, bats possess adaptations unique among mammals including powered flight, unexpected longevity, and extraordinary immunity. Some of the molecular mechanisms underlying these unique adaptations includes DNA repair, metabolism and immunity. However, analyses have been limited to a few divergent lineages, reducing the scope of inferences on gene family evolution across the Order Chiroptera. We conducted an exhaustive comparative genomic study of 37 bat species, one generated in this study, encompassing a large number of lineages, with a particular emphasis on multi-gene family evolution across immune and metabolic genes. In agreement with previous analyses, we found lineage-specific expansions of the APOBEC3 and MHC-I gene families, and loss of the proinflammatory PYHIN gene family. We inferred more than 1,000 gene losses unique to bats, including genes involved in the regulation of inflammasome pathways such as epithelial defence receptors, the natural killer gene complex and the interferon-gamma induced pathway. Gene set enrichment analyses revealed genes lost in bats are involved in defence response against pathogen-associated molecular patterns and damage-associated molecular patterns. Gene family evolution and selection analyses indicate bats have evolved fundamental functional differences compared to other mammals in both innate and adaptive immune system, with the potential to enhance antiviral immune response while dampening inflammatory signalling. In addition, metabolic genes have experienced repeated expansions related to convergent shifts to plant-based diets. Our analyses support the hypothesis that, in tandem with flight, ancestral bats had evolved a unique set of immune adaptations whose functional implications remain to be explored.


Effect of shoulder stabilization on career length in national football league athletes.
Shoulder instability and surgical stabilization are common in college football athletes. The effect of shoulder stabilization during college on the length of an athlete's career in the National Football League (NFL) has not been well examined. Athletes with a history of shoulder stabilization before the NFL combine have a shorter career than do matched controls. Cohort study; Level of evidence, 3. A database containing the injury history and career NFL statistics of athletes from 1987 to 2000 was used to match athletes with a history of shoulder stabilization and no other surgery or significant injury to controls without a history of any previous surgery or significant injury. Athletes were matched by position, year drafted, round drafted, and additional minor injury history. Forty-two athletes with a history of shoulder stabilization were identified and matched with controls. A history of shoulder stabilization significantly reduced the length of career in terms of years (5.2 ± 3.9 vs 6.9 ± 3.6 years; P = .01) and games played (56 ± 53 vs 77 ± 50, P = .03) as compared with controls. By position, linemen and linebackers (20 athletes) with a history of shoulder stabilization had a significantly shorter career in years (4.7 ± 3.8 vs 6.7 ± 3.4 years; P = .049) and games played (51 ± 58 vs 81 ± 48; P = .046) than did controls. Among the other positions (22 athletes), the difference was not statistically significant in this small cohort. A history of shoulder stabilization shortens the expected career of a professional football player, particularly for linemen and linebackers. Further research is warranted to better understand how these injuries and surgeries affect an athlete's career and what can be done to improve the long-term outcome after treatment.


Effects of questionnaire-based diagnosis and training on inter-rater reliability among practitioners of traditional Chinese medicine.
To investigate whether a training process that focused on a questionnaire-based diagnosis in Traditional Chinese Medicine (TCM), and developing diagnostic consensus, would improve the agreement of TCM diagnoses among 10 TCM practitioners evaluating patients with temporomandibular joint disorder (TMJD). Evaluation of a diagnostic training program at the Department of Family and Community Medicine, University of Arizona, Tucson, Arizona, and the Oregon College of Oriental Medicine, Portland, Oregon. Screened participants for a study of TCM for TMJD. PRACTITIONERS: Ten (10) licensed acupuncturists with a minimum of 5 years licensure and education in Chinese herbs. A training session using a questionnaire-based diagnostic form was conducted, followed by waves of diagnostic sessions. Between sessions, practitioners discussed the results of the previous round of participants with a focus on reducing variability in primary diagnosis and severity rating of each diagnosis: 3 waves of 5 patients were assessed by 4 practitioner pairs for a total of 120 diagnoses. At 18 months, practitioners completed a recalibration exercise with a similar format with a total of 32 diagnoses. These diagnoses were then examined with respect to the rate of agreement among the 10 practitioners using inter-rater correlations and kappas. The inter-rater correlation with respect to the TCM diagnoses among the 10 practitioners increased from 0.112 to 0.618 with training. Statistically significant improvements were found between the baseline and 18 month exercises (p < 0.01). Inter-rater reliability of TCM diagnosis may be improved through a training process and a questionnaire-based diagnosis process. The improvements varied by diagnosis, with the greatest congruence among primary and more severe diagnoses. Future TCM studies should consider including calibration training to improve the validity of results.


Investigating Individual Pre-trauma Susceptibility to a PTSD-Like Phenotype in Animals.
Post-Traumatic Stress Disorder (PTSD) is a complex condition that develops after experiencing a severe emotional trauma, with or without physical trauma. There is no known cure and evidence-based treatments, which are effective in reducing symptoms, have low retention rates. It is therefore important, in addition to seeking new therapeutics, to identify ways to reduce the likelihood of developing PTSD. The fact that some, but not all, individuals exposed to the same traumatic event develop PTSD suggests that there is individual susceptibility. Investigating susceptibility and underlying factors will be better guided if there is a coherent framework for such investigations. In this review, we propose that susceptibility is a dynamic state that is comprised of susceptibility factors (before trauma) and sequalae factors (during or after trauma, but before PTSD diagnosis). We define key features of susceptibility and sequalae factors as: (1) they are detectable before trauma (susceptibility factors) or during/shortly after trauma (sequalae factors), (2) they can be manipulated, and (3) manipulation of these factors alters the likelihood of developing PTSD, thus affecting resilience. In this review we stress the importance of investigating susceptibility to PTSD with appropriate animal models, because prospective human studies are expensive and manipulation of susceptibility and sequalae factors for study purposes may not always be feasible. This review also provides a brief overview of a subset of animal models that study PTSD-related behaviors and related alterations in endocrine and brain systems that focus on individual differences, peri- and post-trauma. Attention is drawn to the RISP model (Revealing Individual Susceptibility to a PTSD-like Phenotype) which assesses susceptibility before trauma. Using the RISP model and expression of plasticity-associated immediate early genes, Arc and Homer1a, we have identified impaired hippocampal function as a potential susceptibility factor. We further discuss other putative susceptibility factors and approaches to mitigate them. We assert that this knowledge will guide successful strategies for interventions before, during or shortly after trauma that can decrease the probability of developing PTSD.


Intelligibility, listening difficulty and listening efficiency in auralized classrooms.
In order to obtain an effective speech communication in rooms it is advisable, besides reaching the full intelligibility of words, to minimize the effort paid by the listener in the recognition of the speech material. This twofold requirement is not easily described by the current room acoustic indicators, which are mainly concerned either with a subjective rating by means of word recognition scores or with using listeners' impressions of reported listening difficulties. In this work, the problem is tackled by introducing the concept of "listening efficiency," which is defined as a combination of the accuracy of intelligibility and of the effort spent on achieving this goal. This indicator is here developed, and an application of the former and of the "listening efficiency" is presented in the field of classroom acoustics. Listening tests with pupils and adults were performed and the subsequent statistical analyses indicated several interesting findings. In particular, listening efficiency is able to clearly discriminate between equal intelligibility scores obtained under different acoustical conditions, permitting room acoustics to be tailored for specific groups, such as children.


Decreased bone resorption with soy isoflavone supplementation in postmenopausal women.
To investigate the effect of soy isoflavone supplementation on bone mineral density (BMD) and markers of bone turnover in postmenopausal women. In this randomized, placebo-controlled clinical trial, we used a crossover design to test the effect of soy isoflavone (110 mg/day) (1.3:1.0:0.22 ratio of genistein/daidzein/ glycitein) on bone formation, bone resorption, bone mineral content (BMC), and BMD for 6 months. Postmenopausal women (n = 19), mean age 70.6 +/- 6.3 years and mean time since menopause 19.1 +/- 5.5 years, were given isoflavone supplements for 6 months. There was a 37% decrease in urinary concentrations of type 1 collagen alpha1-chain helical peptide (HP), a marker of bone resorption, during the isoflavone supplementation compared with baseline (p < 0.05) and a significant difference in mean (SE) HP excretion levels when isoflavone was compared with placebo (43.4 +/- 5.2 vs. 56.3 +/- 7.2 microg/mmol creatinine [cr], p < 0.05). With isoflavone supplementation, mean spine BMD at L2 and L3 was significantly greater when treatment was compared with control, with a difference between means of 0.03 +/- 0.04 g and 0.03 +/- 0.04 g (p < 0.05), respectively. There were nonsignificant increases from baseline for total spine BMC (3.5%), total spine BMD (1%), total hip BMC (3.6%), and total hip BMD (1.3%) with the isoflavone treatment. Soy isoflavone, in isolated form, was effective in this study to significantly decrease bone resorption in postmenopausal women. Further investigation needs to be done to evaluate the long-term effects of soy isoflavone on bone mass and fracture risk.


Automatic estimation and removal of noise from a single image.
Image denoising algorithms often assume an additive white Gaussian noise (AWGN) process that is independent of the actual RGB values. Such approaches are not fully automatic and cannot effectively remove color noise produced by todays CCD digital camera. In this paper, we propose a unified framework for two tasks: automatic estimation and removal of color noise from a single image using piecewise smooth image models. We introduce the noise level function (NLF), which is a continuous function describing the noise level as a function of image brightness. We then estimate an upper bound of the real noise level function by fitting a lower envelope to the standard deviations of per-segment image variances. For denoising, the chrominance of color noise is significantly removed by projecting pixel values onto a line fit to the RGB values in each segment. Then, a Gaussian conditional random field (GCRF) is constructed to obtain the underlying clean image from the noisy input. Extensive experiments are conducted to test the proposed algorithm, which is shown to outperform state-of-the-art denoising algorithms.


Antimitochondrial antibody heterogeneity and the xenobiotic etiology of primary biliary cirrhosis.
Antimitochondrial antibodies (AMAs) directed against the lipoyl domain of the E2 subunit of pyruvate dehydrogenase (PDC-E2) are detected in 95% of patients with primary biliary cirrhosis (PBC) and are present before the onset of clinical disease. The recent demonstration that AMAs recognize xenobiotic modified PDC-E2 with higher titers than native PDC-E2 raises the possibility that the earliest events involved in loss of tolerance are related to xenobiotic modification. We hypothesized that reactivity to such xenobiotics would be predominantly immunoglobulin M (IgM) and using sera from a large cohort of PBC patients and controls (n = 516), we examined in detail sera reactivity against either 6,8-bis(acetylthio) octanoic acid (SAc)-conjugated bovine serum albumin (BSA), recombinant PDC-E2 (rPDC-E2) or BSA alone. Further, we also defined the relative specificity to the SAc moiety using inhibition enzyme-linked immunosorbent assay (ELISA); SAc conjugate and rPDC-E2-specific affinity-purified antibodies were also examined for antigen specificity, isotype, and crossreactivity. Reactivity to SAc conjugates is predominantly IgM; such reactivity reflects a footprint of previous xenobiotic exposure. Indeed, this observation is supported by both direct binding, crossreactivity, and inhibition studies. In both early and late-stage PBC, the predominant Ig isotype to SAc is IgM, with titers higher with advanced stage disease. We also note that there was a higher level of IgM reactivity to SAc than to rPDC-E2 in early-stage versus late-stage PBC. Interestingly, this finding is particularly significant in light of the structural similarity between SAc and the reduced form of lipoic acid, a step which is similar to the normal physiological oxidation of lipoic acid. Specific modifications of the disulfide bond within the lipoic-acid-conjugated PDC-E2 moiety, i.e., by an electrophilic agent renders PDC-E2 immunogenic in a genetically susceptible host.


Separation and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex.
Saponins were purified from Quillaja saponaria Molina bark by silica and reverse phase chromatography. The resulting purified saponins were tested for adjuvant activity in mice. Several distinct saponins, designated QS-7, QS-17, QS-18, and QS-21, were demonstrated to boost antibody levels by 100-fold or more when used in mouse immunizations with the Ag BSA and beef liver cytochrome b5. These purified saponins increased titers in all major IgG subclasses. To determine optimal dose in mice for adjuvant response, QS-7 and QS-21 were tested in a dose-response study in intradermal immunization with BSA in mice; for both of these purified saponins, adjuvant response (determined by stimulation of ELISA titers to BSA) neared maximum at doses of 5 micrograms and was shown to plateau up to the highest dose tested, 80 micrograms. These purified saponins vary considerably in their toxicity, as assessed by lethality in mice; the main component, QS-18, being the most toxic. Saponins QS-7 and QS-21 showed no or very low toxicity in mice, respectively. None of these saponins stimulated production of reaginic antibodies. The monosaccharide composition of these saponins showed similar but distinct compositions with all four containing fucose, xylose, galactose and glucuronic acid. Predominant differences were observed in the quantities of rhamnose, arabinose, and glucose. Monomer m.w. (determined by size exclusion HPLC) were determined to range from 1800 to 2200.


Modulation of the time course of fast EPSCs and glutamate channel kinetics by aniracetam.
It is generally accepted that glutamate serves as the neurotransmitter at most excitatory synapses in the mammalian central nervous system (CNS). Synaptic release of glutamate may trigger a fast and a slow excitatory postsynaptic current (EPSC). The slow EPSC is mediated by N-methyl-D-aspartate (NMDA) receptor channels, whereas the fast EPSC is mediated by non-NMDA receptor channels. The nootropic agent aniracetam selectively and reversibly slows the desensitization kinetics of non-NMDA channels and lengthens their single-channel open times. Antiracetam also modulates the kinetics of the fast EPSC in a manner that mirrors its action on the kinetics of the non-NMDA channels. These results support the hypothesis that the properties of the non-NMDA glutamate channels rather than the rate of neurotransmitter clearance are the primary determinants of the kinetics of the fast EPSC in the mammalian CNS.


Exploring the mechanism of F282L mutation-caused constitutive activity of GPCR by a computational study.
G-protein-coupled receptors (GPCRs) are important drug targets and generally activated by ligands. However, some experiments found that GPCRs also give rise to constitutive activity through some mutations (viz., CAM), which are usually associated with different kinds of diseases. However, the mechanisms of CAMs and their roles in interactions with drug-ligands are unclear in experiments. Herein, we used microsecond molecular dynamics simulations to study the effect of one important F282L mutation on β2AR in order to address the questions above. With the aid of principle component and correlation analysis, our results revealed that the F282L mutation could increase the instability of the overall structure, increase the dramatic fluctuations of NPxxY and extracellular loops, and decrease restraint of the helices through weakening interhelical H-bonding and correlations between residues, which could partly contribute to the constitutive activity reported by the experiments. The observations from the protein structure network (PSN) analysis indicate that the mutant exhibits less information flow than the wild β2AR and weakens the role of TM5 and TM6 in the signal transmission, but it enhances the impact of TM3 on the orthosteric pathway and TM4 on the allosteric one. In addition, the results from the virtual screening reveal that the mutant prefers to select agonists rather than antagonists, similar to the active state but opposite of the inactive state, further confirming that the F282L mutation advances the activation of β2AR. Our observations provide valuable information for understanding the mechanism of the mutation-caused constitutive activity of GPCR and related drug-design.


Frequency domain depth filtering of integral imaging.
A novel technique for depth filtering of integral imaging is proposed. Integral imaging captures spatio-angular distribution of the light rays which delivers three-dimensional information of the object scene. The proposed method performs filtering operation in the frequency domain of the captured spatio-angular light ray distribution, achieving depth selective reconstruction. Grating projection further enhances the depth discrimination performance. The principle is verified experimentally.