Antibacterial and Anti-inflammatory Potential of Morus alba Stem Extract.
Periodontitis, a chronic inflammatory disease, is the leading cause of tooth loss in adults. Evidence for the anti inflammatory activity of M. alba Stem Extract (MSE) in periodontal disease is limited. The study aimed to investigate the inhibitory effect of MSE on the growth of periodontopathic bacteria and expression of interleukin (IL)-6 and IL-8 in Porphyromonas gingivalis Lipopolysaccharide (LPS)-stimulated human Periodontal Ligament (hPDL) fibroblasts. The antimicrobial activities of MSE were tested against P. gingivalis and Actinobacillus actinomycetemcomitans by the disk diffusion, the minimum inhibitory concentration and the minimal bactericidal concentration methods. Cytotoxicity of P. gingivalis LPS and MSE on hPDL fibroblasts was determined by MTS assay. The expression of cytokines (IL-6 and IL-8) mRNA and proteins in hPDL fibroblasts was measured using the reverse transcription-qPCR and enzyme-linked immunosorbent assay, respectively. MSE exhibited antibacterial activities against P. gingivalis and A. actinomycetemcomitans with the zones of inhibition of 10.00 ± 0.33 mm and 17.33 ± 0.58 mm, respectively. MIC and MBC values for MSE against P. gingivalis were 62.5 μg/ml. The MIC and MBC values against A. actinomycetemcomitans were 250 μg/mL and 500 μg/ml, respectively. P. gingivalis LPS was shown to mediate the expression of pro-inflammatory cytokines in hPDL fibroblasts. However, treatment with MSE concentrations of 2.5 and 5.0 μg/ml significantly suppressed P. gingivalis LPS-induced IL-6 and IL-8 mRNA and protein expression (p< 0.05). The present study demonstrates that MSE has antibacterial activity against two putative periodontal pathogens. MSE suppressed IL-6 and IL-8 expression in P. gingivalis LPS-stimulated hPDL fibroblasts, indicating a possible anti-inflammatory effect. Thus, it is a potential adjunctive agent for the treatment of periodontitis.


Educational Needs Associated with the Level of Complication and Comparative Risk Perceptions in People with Type 2 Diabetes.
This study aimed to identify the educational needs of people with type 2 diabetes according to risk perceptions and the level of severity of complications. There were 177 study participants who were outpatients of the internal medicine department at a university hospital located in the Republic of Korea, who consented to participate in the survey from December 10, 2016 to February 10, 2017. The data were analyzed using descriptive statistics, Pearson correlation, ANOVA with post-hoc comparison, and multiple regression analysis. Type 2 diabetes complications were classified into 3 groups: no complications, common complications, and severe complications. There were statistically significant positive correlations between educational needs and comparative risk perceptions, and the level of complication and comparative risk perception. Multiple regression analysis revealed that the factor predicting educational needs of type 2 diabetes people was their comparative risk perceptions, rather than the severity of diabetes complications or sociodemographic variables. Since risk perception is the factor that indicates the educational needs of people with type 2 diabetes, there is a need to explore factors which increase risk perception, in order to meet educational needs. The findings suggest that a more specific and individualized educational program, which focuses on each person's risk perceptions, should be developed.


Presynaptic adenosine A1 receptors regulate retinohypothalamic neurotransmission in the hamster suprachiasmatic nucleus.
Adenosine has been implicated as a modulator of retinohypothalamic neurotransmission in the suprachiasmatic nucleus (SCN), the seat of the light-entrainable circadian clock in mammals. Intracellular recordings were made from SCN neurons in slices of hamster hypothalamus using the in situ whole-cell patch clamp method. A monosynaptic, glutamatergic, excitatory postsynaptic current (EPSC) was evoked by stimulation of the optic nerve. The EPSC was blocked by bath application of the adenosine A(1) receptor agonist cyclohexyladenosine (CHA) in a dose-dependent manner with a half-maximal concentration of 1.7 microM. The block of EPSC amplitude by CHA was antagonized by concurrent application of the adenosine A(1) receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX). The adenosine A(2A) receptor agonist CGS21680 was ineffective in attenuating the EPSC at concentrations up to 50 microM. Trains of four consecutive stimuli at 25 ms intervals usually depressed the EPSC amplitude. However, after application of CHA, consecutive responses displayed facilitation of EPSC amplitude. The induction of facilitation by CHA suggested a presynaptic mechanism of action. After application of CHA, the frequency of spontaneous EPSCs declined substantially, while their amplitude distribution was unchanged or slightly reduced, again suggesting a mainly presynaptic site of action for CHA. Application of glutamate by brief pressure ejection evoked a long-lasting inward current that was unaffected by CHA at concentrations sufficient to reduce the evoked EPSC amplitude substantially (1 to 5 microM), suggesting that postsynaptic glutamate receptor-gated currents were unaffected by the drug. Taken together, these observations indicate that CHA inhibits optic nerve-evoked EPSCs in SCN neurons by a predominantly presynaptic mechanism.


Aggregation of Au(i)-complexes on amorphous substrates governed by aurophilicity.
In single crystals of 2-naphthylisonitrile-gold(i)-halide (halide = Cl, Br, I) complexes, AuAu distances are found to be significantly shorter than twice the van der Waals radius, indicating attractive interactions between gold atoms in adjacent molecules. In the particular case of the studied 2-naphthylisonitrile-gold(i) complexes, homodimers are the common structural motifs, in which the linearly coordinated gold exhibits a crossed swords arrangement with the Au atoms of two molecules being at the intersection point. The crossed swords motif is preserved upon physical vapour deposition of both the chlorine and bromine derivatives on amorphous substrates like glass and glassy carbon. The determined activation energies of desorption for the chlorine (0.9 eV) and the bromine (1.2 eV) derivative are comparable to that of unsubstituted naphthalene. Using X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and ion scattering (RBS), we confirmed the chemical integrity of the molecules in thin films and revealed the orientation of the crossed swords dimers with respect to the substrate surface.


Ancestral divergence, genome diversification, and phylogeographic variation in subpopulations of sorbitol-negative, beta-glucuronidase-negative enterohemorrhagic Escherichia coli O157.
The O157:H7 lineage of enterohemorrhagic Escherichia coli is a geographically disseminated complex of highly related genotypes that share common ancestry. The common clone that is found worldwide carries several markers of events in its evolution, including markers for acquisition of virulence genes and loss of physiological characteristics, such as sorbitol fermentation ability and beta-glucuronidase production. Populations of variants that are distinct with respect to motility and the sorbitol and beta-glucuronidase markers appear to have diverged at several points along the inferred evolutionary pathway. In addition to these variants, distinct subpopulations of the contemporary non-sorbitol-fermenting, beta-glucuronidase-negative O157:H7 clone were recently detected among bovine and human clinical isolates in the United States by using high-resolution genome comparison. In order to determine if these recently described subpopulations were derived from a regional or ancestral divergence event, we used octamer-based genome scanning, marker sorting, and DNA sequence analysis to examine their phylogenetic relationship to populations of non-sorbitol-fermenting, beta-glucuronidase negative O157:H7 and O157:H- strains from Australia. The inferred phylogeny is consistent with the hypothesis that subpopulations on each continent resulted from geographic spread of an ancestral divergence event and subsequent expansion of distinct subpopulations. Marker sorting and DNA sequence analyses identified sets of monophyletic markers consistent with the pattern of divergence and demonstrated that phylogeographic variation occurred through emergence of regional subclones and concentration of regional polymorphisms among distinct subpopulations. DNA sequence analysis of representative polyphyletic markers showed that genome diversity accrued through random drift and bacteriophage-mediated events.


Synesthetic congruency modulates the temporal ventriloquism effect.
People sometimes find it easier to judge the temporal order in which two visual stimuli have been presented if one tone is presented before the first visual stimulus and a second tone is presented after the second visual stimulus. This enhancement of people's visual temporal sensitivity has been attributed to the temporal ventriloquism of the visual stimuli toward the temporally proximate sounds, resulting in an expansion of the perceived interval between the two visual events. In the present study, we demonstrate that the synesthetic congruency between the auditory and visual stimuli (in particular, between the relative pitch of the sounds and the relative size of the visual stimuli) can modulate the magnitude of this multisensory integration effect: The auditory capture of vision is larger for pairs of auditory and visual stimuli that are synesthetically congruent than for pairs of stimuli that are synesthetically incongruent, as reflected by participants' increased sensitivity in discriminating the temporal order of the visual stimuli. These results provide the first evidence that multisensory temporal integration can be affected by the synesthetic congruency between the auditory and visual stimuli that happen to be presented.


Comprehensive predictive biomarker analysis for MEK inhibitor GSK1120212.
The MEK1 and MEK2 inhibitor GSK1120212 is currently in phase II/III clinical development. To identify predictive biomarkers, sensitivity to GSK1120212 was profiled for 218 solid tumor cell lines and 81 hematologic malignancy cell lines. For solid tumors, RAF/RAS mutation was a strong predictor of sensitivity. Among RAF/RAS mutant lines, co-occurring PIK3CA/PTEN mutations conferred a cytostatic response instead of a cytotoxic response for colon cancer cells that have the biggest representation of the comutations. Among KRAS mutant cell lines, transcriptomics analysis showed that cell lines with an expression pattern suggestive of epithelial-to-mesenchymal transition were less sensitive to GSK1120212. In addition, a proportion of cell lines from certain tissue types not known to carry frequent RAF/RAS mutations also seemed to be sensitive to GSK1120212. Among these were breast cancer cell lines, with triple negative breast cancer cell lines being more sensitive than cell lines from other breast cancer subtypes. We identified a single gene DUSP6, whose expression was associated with sensitivity to GSK1120212 and lack of expression associated with resistance irrelevant of RAF/RAS status. Among hematologic cell lines, acute myeloid leukemia and chronic myeloid leukemia cell lines were particularly sensitive. Overall, this comprehensive predictive biomarker analysis identified additional efficacy biomarkers for GSK1120212 in RAF/RAS mutant solid tumors and expanded the indication for GSK1120212 to patients who could benefit from this therapy despite the RAF/RAS wild-type status of their tumors.


The WHO antenatal care randomised controlled trial: rationale and study design.
The World Health Organisation and collaborating institutions in developing countries are conducting a multicentre randomised controlled trial to evaluate a new antenatal care (ANC) programme, consisting of tests, clinical procedures and follow-up actions scientifically demonstrated to be effective in improving maternal and newborn outcomes. These activities are distributed, for practical reasons, over four visits during the course of pregnancy and are aimed at achieving predetermined goals. The study is taking place in four countries, Argentina, Cuba, Saudi Arabia and Thailand. Recruitment of study subjects started on 1 May 1996. All 53 ANC clinical units had been enrolled by December 1996. Clinics in each country were randomly allocated (cluster randomisation) to provide either the new programme or the traditional programme currently in use. Approximately 24,000 women presenting for ANC at these clinics over an average period of 18 months will have been recruited. As women attending the control clinics receive the 'best standard treatment' as currently offered in these clinics, individual informed consent is requested only from women attending the intervention clinics. Authorities of the corresponding health districts and all participating clinics have provided written institutional informed consent before randomisation. The primary outcome of the trial in relation to maternal conditions is the rate of a morbidity indicator index, defined as the presence of at least one of the following conditions for which ANC is relevant: (a) pre-eclampsia or eclampsia during pregnancy or within 24 h of delivery; (b) postpartum anaemia (haemoglobin < 90 g/L); or (c) severe urinary tract infection/pyelonephritis, defined as an episode requiring antibiotic treatment and/or hospitalisation. The primary fetal outcome is the rate of low birthweight (< 2500 g). Adverse maternal and fetal outcomes are expected for approximately 10% of the control group. Several maternal and perinatal secondary outcomes are also considered. A comprehensive cost-effectiveness analysis and women's and providers' satisfaction evaluation are performed concurrently with the trial. Health-care programmes should be rigorously evaluated by randomised controlled trials, which are feasible in developing countries and should be conducted before introducing new treatments or health interventions.


Corporate health benefits and the indexing of the personal income tax.
This note focuses on the role of the personal income tax in reducing the effective price of health care benefits. Tax-bracket creep is shown to provide a cushion that absorbs relatively large increases in health benefit costs, thus reducing the impetus for employer initiatives to control health care costs. It is hypothesized that the Economic Recovery Tax Act of 1981, with its provision for the indexing of tax brackets, will increase employer concern, and may therefore spur the development of effective employer initiatives to reduce the costs of health benefits.


Status of High latitude precipitation estimates from observations and reanalyses.
An intercomparison of high-latitude precipitation characteristics from observation-based and reanalysis products is performed. In particular the precipitation products from CloudSat provide an independent assessment to other widely used products, these being the observationally-based GPCP, GPCC and CMAP products and the ERA-Interim, MERRA and NCEP-DOE R2 reanalyses. Seasonal and annual total precipitation in both hemispheres poleward of 55° latitude is considered in all products, and CloudSat is used to assess intensity and frequency of precipitation occurrence by phase, defined as rain, snow or mixed phase. Furthermore, an independent estimate of snow accumulation during the cold season was calculated from the Gravity Recovery and Climate Experiment (GRACE). The intercomparison is performed for the 2007-2010 period when CloudSat was fully operational. It is found that ERA- Interim and MERRA are broadly similar, agreeing more closely with CloudSat over oceans. ERA-Interim also agrees well with CloudSat estimates of snowfall over Antarctica where total snowfall from GPCP and CloudSat is almost identical. A number of disagreements on regional or seasonal scales are identified: CMAP reports much lower ocean precipitation relative to other products, NCEP-DOE R2 reports much higher summer precipitation over northern hemisphere land, GPCP reports much higher snowfall over Eurasia, and CloudSat overestimates precipitation over Greenland, likely due to mischaracterization of rain and mixed-phase precipitation. These outliers are likely unrealistic for these specific regions and time periods. These estimates from observations and reanalyses provide useful insights for diagnostic assessment of precipitation products in high latitudes, quantifying the current uncertainties, improving the products, and establishing a benchmark for assessment of climate models.


Structural Characterization of Lecithin-Stabilized Tetracosane Lipid Nanoparticles. Part I: Emulsions.
The structure of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)-stabilized colloidal tetracosane emulsions was investigated by photon correlation spectroscopy and small-angle X-ray and neutron scattering, using emulsions with different neutron scattering contrasts. Special emphasis was placed on the structure of the DMPC stabilizer layer covering the emulsion droplets. A monolayer, structurally similar to a half DMPC bilayer, with a thickness of 16 Å is found. Thereby, the phosphocholine headgroups arrange flat at the oil-water interface. A deep penetration of the tetracosane oil into the stabilizer layer can be ruled out.


Soft-x-ray hollow fiber optics with inner metal coating.
A glass capillary with an inner metal coating is proposed to be used as soft-x-ray fiber optics in medical applications. Based on the results of theoretical calculations, nickel was chosen as the coating material for x rays radiated from a conventional x-ray tube. A nickel-coated capillary was fabricated by electroless deposition, and focusing and collimating effects were observed from measurements of the transmission efficiency of soft x rays. The transmission of a nickel-coated capillary with an inner diameter of 0.53 mm and a length of 300 mm was 10%, which is approximately double that of an uncoated glass capillary.


Holmium:YAG laser ablation combined intraurethral fluorouracil perfusion as treatment option for intraurethral Condyloma acuminata in men.
Intraurethral condylomata acuminata (CA) is caused by human papilloma virus (HPV) infection which is transmitted by close physical and sexual contact. CA is often difficult to cure. There is limited research on the treatment of the patients with intraurethral CA. Here, we have reviewed our experiences on the treatment of intraurethral condylomatous with Holmium:YAG Laser ablation. A new and convenient mean of administering fluorouracil and lidocaine for the treatment of intraurethral condyloma acuminata is discussed. This study aimed to evaluate the experience and efficacy of Holmium:YAG Laser ablation with ureteroscopy and local administration of fluorouracil in the treatment of patients with intraurethral CA. The effects were investigated based on the rate of cure and relapse and the incidence of complications. The study included patients with intraurethral condylomatous who had undergone Holmium:YAG Laser ablation and intraurethral perfusion of fluorouracil. From May 2005 to October 2008, 25 patients (mean age 31.3 years, 19-63 years) with cystourethroscopy confirmed extensive lesions at the anterior urethra and biopsy of the lesions was compatible with condyloma acuminata. They all underwent Holmium:YAG Laser ablation with a transurethral Wolf 8/9.8 Fr rigid ureteroscope. And a week later, the patients initially accepted intraurethral installation of the mixture containing 1% fluorouracil and 1% tetracaine hydrochloride gel (lubricating jelly) in a volume of 20 mL. This mixture was given intraurethrally once weekly, and tip of the penis was clamped immediately to close the urethral meatus after administration by using an occlusive penile clamp and was retained for 20 minutes. Six treatments were given initially and after six weeks of rest, another cycle of six weekly treatments was given. Ureteroscopic Holmium laser ablation was successfully performed in all patients with multifocal intraurethral CA. Mean CA warts body size was 3 mm (2-8) in diameter. Mean operative time was 22.8 minutes (range 13-41). No major intraoperative complications occurred. Intraurethral installation was well tolerated, although six patients complained occasional urethral pain while urinating. Three relapses in a 2-5 weeks of follow-up underwent repeat holmium laser ablation and installation of the fluorouracil mixture. In an average of six months of follow-up, the patients have no ureteral stricture or relapse of the CA. The results of this study suggest that holmium:YAG laser ablation of the intraurethral CA combined with intraurethral perfusion of 5-fluorouracil and tetracaine hydrochloride gel mixture is an effective and safer therapy with a lower relapse rate for treatment of intraurethral CA.


Influence of reactive species on the modification of biomolecules generated from the soft plasma.
Plasma medicine is an upcoming research area that has attracted the scientists to explore more deeply the utility of plasma. So, apart from the treating biomaterials and tissues with plasma, we have studied the effect of soft plasma with different feeding gases such as Air, N2 and Ar on modification of biomolecules. Hence, in this work we have used the soft plasma on biomolecules such as proteins ((Hemoglobin (Hb) and Myoglobin (Mb)), calf thymus DNA and amino acids. The structural changes or structural modification of proteins and DNA have been studied using circular dichroism (CD), fluorescence spectroscopy, protein oxidation test, gel electrophoresis, UV-vis spectroscopy, dynamic light scattering (DLS) and 1D NMR, while Liquid Chromatograph/Capillary Electrophoresis-Mass Spectrometer (LC/CE-MS) based on qualitative and quantitative bio-analysis have been used to study the modification of amino acids. Further, the thermal analysis of the protein has been studied with differential scanning calorimetry (DSC) and CD. Additionally, we have performed docking studies of H2O2 with Hb and Mb, which reveals that H2O2 molecules preferably attack the amino acids near heme group. We have also shown that N2 gas plasma has strong deformation action on biomolecules and compared to other gases plasma.


Cloning of complementary DNA encoding a functional human interleukin-8 receptor.
Interleukin-8 (IL-8) is an inflammatory cytokine that activates neutrophil chemotaxis, degranulation, and the respiratory burst. Neutrophils express receptors for IL-8 that are coupled to guanine nucleotide-binding proteins (G proteins); binding of IL-8 to its receptor induces the mobilization of intracellular calcium stores. A cDNA clone from HL-60 neutrophils, designated p2, has now been isolated that encodes a human IL-8 receptor. When p2 is expressed in oocytes from Xenopus laevis, the oocytes bind 125I-labeled IL-8 specifically and respond to IL-8 by mobilizing calcium stores with an EC50 of 20 nM. This IL-8 receptor has 77% amino acid identity with a second human neutrophil receptor isotype that binds IL-8 with higher affinity. It also exhibits 69% amino acid identity with a protein reported to be an N-formyl peptide receptor from rabbit neutrophils, but less than 30% identity with all other known G protein-coupled receptors, including the human N-formyl peptide receptor.


Structure, dynamics, and reactivity in hemoglobin.
The static structure of hemoglobin and its functional properties are very well characterized. It is still not known how energy is stored and used within the structure of the protein to promote function and functional diversity. An essential part of this question is understanding the mechanism through which the overall protein structure (quaternary structure) couples to the local environment about the oxygen binding sites. Time-resolved resonance Raman spectroscopy has been used to probe the vibrational degrees of the freedom of the binding site as a function of protein structure. Comparison of the spectra from both equilibrium and transient forms of deoxy hemoglobin from a variety of mammalian, reptilian, and fish hemoglobins reveals that for each quaternary structure there exist two tertiary states stabilized by the presence or absence of an iron-bound ligand. Pulse-probe Raman experiments show that for photodissociated, ligated hemoglobins the local tertiary structure relaxes at a solution-dependent rate extending from tens of nanoseconds to microseconds. In this local environment, the linkage between the iron and the proximal histidine proves to be the single observed structural feature that responds in a systematic and substantial manner to structural changes in the protein. The additional finding of a correlation between the frequency of the iron-proximal histidine stretching motion (nu Fe-His) and various parameters of ligand reactivity, including geminate recombination, implicates the associated localized structural element in the mechanism of protein control of ligand binding. On the basis of these and related finds, a model is presented to account for both coarse and fine control of ligand binding by the protein structure.


Prevalence and burden of hepatitis D virus infection in the global population: a systematic review and meta-analysis.
Hepatitis D virus (HDV) is a defective virus that completes its life cycle only with hepatitis B virus (HBV). The HBV with HDV super-infection has been considered as one of the most severe forms of the chronic viral hepatitis. However, there is a scarcity of data on the global burden of HDV infection. We searched PubMed, Embase, Cochrane Library and China Knowledge Resource Integrated databases from 1 January 1977 to 31 December 2016. We included studies with a minimum sample size of 50 patients. Our study analysed data from a total of 40 million individuals to estimate the prevalence of HDV by using Der-Simonian Laird random-effects model. The data were further categorised according to risk factors. From a total of 2717 initially identified studies, only 182 articles from 61 countries and regions met the final inclusion criteria. The overall prevalence of HDV was 0.98% (95% CI 0.61 to 1.42). In HBsAg-positive population, HDV pooled prevalence was 14.57% (95% CI 12.93 to 16.27): Seroprevalence was 10.58% (95% CI 9.14 to 12.11) in mixed population without risk factors of intravenous drug use (IVDU) and high-risk sexual behaviour (HRSB). It was 37.57% (95% CI 29.30 to 46.20) in the IVDU population and 17.01% (95% CI 10.69 to 24.34) in HRSB population. We found that approximately 10.58% HBsAg carriers (without IVDU and HRSB) were coinfected with HDV, which is twofold of what has been estimated before. We also noted a substantially higher HDV prevalence in the IVDU and HRSB population. Our study highlights the need for increased focus on the routine HDV screening and rigorous implementation of HBV vaccine programme.


Detection and Discovery of Coronaviruses in Wild Bird Populations.
Wild birds are natural hosts of multiple microbial agents, including a wide diversity of coronaviruses. Here we describe a pan-Coronavirus detection RT-PCR method to identify those viruses regardless of the coronavirus genus or nature of the specimen. We also describe a protocol using high-throughput sequencing technologies to obtain their entire genome, which overcomes the inherent difficulties of wild bird coronavirus sequencing, that is, their genetic diversity and the lack of virus isolation methods.


Potentially human-pathogenic Escherichia coli O26 in Norwegian sheep flocks.
A national survey of Escherichia coli O26 in Norwegian sheep flocks was conducted, using fecal samples to determine the prevalence. In total, 491 flocks were tested, and E. coli O26 was detected in 17.9% of the flocks. One hundred forty-two E. coli O26 isolates were examined for flagellar antigens (H typing) and four virulence genes, including stx and eae, to identify possible Shiga toxin-producing E. coli (STEC) and enteropathogenic E. coli (EPEC). Most isolates (129 out of 142) were identified as E. coli O26:H11. They possessed eae and may have potential as human pathogens, although only a small fraction were identified as STEC O26:H11, giving a prevalence in sheep flocks of only 0.8%. Correspondingly, the sheep flock prevalence of atypical EPEC (aEPEC) O26:H11 was surprisingly high (15.9%). The genetic relationship between the E. coli O26:H11 isolates was investigated by pulsed-field gel electrophoresis (PFGE) and multilocus variable number tandem repeat analysis (MLVA), identifying 63 distinct PFGE profiles and 22 MLVA profiles. Although the MLVA protocol was less discriminatory than PFGE and a few cases of disagreement were observed, comparison by partition mapping showed an overall good accordance between the two methods. A close relationship between a few isolates of aEPEC O26:H11 and STEC O26:H11 was identified, but all the E. coli O26:H11 isolates should be considered potentially pathogenic to humans. The present study consisted of a representative sampling of sheep flocks from all parts of Norway. This is the first large survey of sheep flocks focusing on E. coli O26 in general, including results of STEC, aEPEC, and nonpathogenic isolates.


Thoracoscopic resection of symptomatic herniated thoracic discs: clinical results in 121 patients.
Retrospective review of a prospectively maintained surgical database. To report the indications, surgical procedures performed, and outcomes from the largest series of thoracoscopically treated herniated thoracic discs (HTDs). We also compared approach-related complications with an unmatched cohort undergoing thoracotomy for HTD. Symptomatic HTDs are rare, and their surgical management is technically challenging. A prospectively maintained surgical database of all patients undergoing surgery for symptomatic HTDs by the senior author (blinded for review) was reviewed. As needed, the database was supplemented with hospital and clinic charts and telephone conversations with patients. A triportal method of thoracoscopic discectomy was performed in all cases. Between 1994 and 2008, 121 patients underwent 125 thoracoscopic-assisted operations for 139 HTDs. Their mean age at surgery was 46.6 years. Indications for thoracoscopic resection currently include small symptomatic disc, anterior location, nonmorbidly obese patient, favorable chest anatomy, and T4-T11 location. Symptom duration averaged 32 months. Radiculopathy was the most common presentation, followed by myelopathy and pain (radiculopathic or back). The mean hospital stay was 4.8 days. Chest tubes remained in place for a mean of 3.2 days. At a mean follow-up of 2.4 years, myelopathy, radiculopathy, and back pain had resolved or improved at a rate of 91.1%, 97.6%, and 86.5%, respectively. Patients reported worsening in 0%, 1.2%, and 0% of cases, respectively. Most patients (97.4%) would be willing to undergo the operation again. The complication rate was acceptable. Patients undergoing thoracoscopic excision had less approach-related morbidity than an unmatched cohort undergoing excision using thoracotomy. Thoracoscopic-assisted microsurgical resection is a safe, effective, and minimally invasive method of treating symptomatic HTDs in appropriately selected patients. The symptoms of most patients improve or resolve with minimal morbidity.


Intake of coffee, decaffeinated coffee, or tea does not affect risk for pancreatic cancer: results from the European Prospective Investigation into Nutrition and Cancer Study.
Few modifiable risk factors have been implicated in the etiology of pancreatic cancer. There is little evidence for the effects of caffeinated coffee, decaffeinated coffee, or tea intake on risk of pancreatic cancer. We investigated the association of total coffee, caffeinated coffee, decaffeinated coffee, and tea consumption with risk of pancreatic cancer. This study was conducted within the European Prospective Investigation into Nutrition and Cancer cohort, comprising male and female participants from 10 European countries. Between 1992 and 2000, there were 477,312 participants without cancer who completed a dietary questionnaire and were followed up to determine pancreatic cancer incidence. Coffee and tea intake was calibrated with a 24-hour dietary recall. Adjusted hazard ratios (HRs) were computed using multivariable Cox regression. During a mean follow-up period of 11.6 y, 865 first incidences of pancreatic cancers were reported. When divided into fourths, neither total intake of coffee (HR, 1.03; 95% confidence interval [CI], 0.83-1.27; high vs low intake), decaffeinated coffee (HR, 1.12; 95% CI, 0.76-1.63; high vs low intake), nor tea were associated with risk of pancreatic cancer (HR, 1.22, 95% CI, 0.95-1.56; high vs low intake). Moderately low intake of caffeinated coffee was associated with an increased risk of pancreatic cancer (HR, 1.33; 95% CI, 1.02-1.74), compared with low intake. However, no graded dose response was observed, and the association attenuated after restriction to histologically confirmed pancreatic cancers. Based on an analysis of data from the European Prospective Investigation into Nutrition and Cancer cohort, total coffee, decaffeinated coffee, and tea consumption are not related to the risk of pancreatic cancer.


Vps9p is a guanine nucleotide exchange factor involved in vesicle-mediated vacuolar protein transport.
Vacuolar protein sorting (vps) mutants of Saccharomyces cerevisiae missort and secrete vacuolar hydrolases. The gene affected in one of these mutants, VPS21, encodes a member of the Sec4/Ypt/Rab family of small GTPases. Rab proteins play an essential role in vesicle-mediated protein transport. Using both yeast two-hybrid assays and chemical cross-linking, we have identified another VPS gene product, Vps9p, that preferentially interacts with a mutant form of Vps21p-S21N that binds GDP but not GTP. In vitro purified Vps9p was found to stimulate GDP release from Vps21p in a dose-dependent manner. Vps9p also stimulated GTP association as a result of facilitated GDP release. However, Vps9p did not stimulate guanine nucleotide exchange of GTP-bound Vps21p or GTP hydrolysis. We tested the ability of Vps9p to stimulate the intrinsic guanine nucleotide exchange activity of Rab5, which is a mammalian sequence homologue of Vps21p, and Ypt7p, which is another yeast Rab protein involved in vacuolar protein transport. Rab5, but not Ypt7p was responsive to Vps9p, which indicates that Vps9p recognizes sequence variation among Rab proteins. We conclude that Vps9p is a novel guanine nucleotide exchange factor that is specific for Vps21p/Rab5. Since there are no obvious Vps9p sequence homologues in yeast, Vps9p may also possess unique regulatory functions required for vacuolar protein transport.


Follow-up care of patients treated for breast cancer: a structured review.
Breast cancer follow-up services vary, with little evidence to support which practice is best. A systematic review methodology was employed to identify and integrate primary research on the effectiveness of follow-up services. From 4418 articles identified by searches, 38 were eligible for review inclusion. Data were not sufficiently homogenous to integrate statistically, however the following patterns of findings were observed: patient survival and quality of life were not affected by intensity of follow-up or location of care; patients held positive attitudes towards follow-up but psychological distress was consistently high regardless of location of services; few studies assessed patient involvement in treatment choices; studies' research quality was poor with inadequate measures of effectiveness or research designs. There is insufficient primary empirical evidence to draw broad conclusions regarding best practice for breast cancer follow-up care in terms of (a) patient involvement in care, (b) reductions in morbidity, and (c) cost effectiveness of service provision.


Variational B-spline level-set: a linear filtering approach for fast deformable model evolution.
In the field of image segmentation, most level-set-based active-contour approaches take advantage of a discrete representation of the associated implicit function. We present in this paper a different formulation where the implicit function is modeled as a continuous parametric function expressed on a B-spline basis. Starting from the active-contour energy functional, we show that this formulation allows us to compute the solution as a restriction of the variational problem on the space spanned by the B-splines. As a consequence, the minimization of the functional is directly obtained in terms of the B-spline coefficients. We also show that each step of this minimization may be expressed through a convolution operation. Because the B-spline functions are separable, this convolution may in turn be performed as a sequence of simple 1-D convolutions, which yields an efficient algorithm. As a further consequence, each step of the level-set evolution may be interpreted as a filtering operation with a B-spline kernel. Such filtering induces an intrinsic smoothing in the algorithm, which can be controlled explicitly via the degree and the scale of the chosen B-spline kernel. We illustrate the behavior of this approach on simulated as well as experimental images from various fields.


Predicting Mortality of Ruptured Abdominal Aortic Aneurysms in the Era of Endovascular Repair.
Previous risk prediction models of mortality after ruptured abdominal aortic aneurysm (rAAA) repair have been limited by imprecision, complexity, or inclusion of variables not available in the preoperative setting. Most importantly, these prediction models have been derived and validated before the adoption of endovascular aneurysm repair (EVAR) as a treatment for rAAA. We sought to derive and validate a new risk-prediction tool using only easily obtainable preoperative variables in patients with rAAA who are being considered for repair in the endovascular era. We used the Vascular Study Group of New England (VSGNE) database to identify all patients who underwent repair of RAAA (2006-2015). Variables were entered into a multivariable logistic regression model to identify independent predictors of 30-day mortality. Linear regression was then used to develop an equation to predict risk of 30-day mortality. During the study period, 649 patients underwent repair of rAAA; of these, 247 (38.1%) underwent EVAR and 402 (61.9%) underwent an open repair. The overall mortality associated with rAAA was 30.7% (open, 33.4% and EVAR, 26.2%). On multivariate modeling, the primary determinants of 30-day mortality were advanced age (>76 vs. ≤76 years, odds ratio [OR] = 2.91 and CI: 2.0-4.24), elevated creatinine (>1.5 mg/dL vs. ≤1.5 mg/dL, OR = 1.57 and CI: 1.05-2.34), and lowest systolic blood pressure (SBP) (BP <70 mm Hg vs. ≥70 mm Hg, OR = 2.65 and CI: 1.79-3.92). The logistic regression model had an area under a c-statistic of 0.69. The corresponding linear model used to provide a point estimate of 30-day mortality (%) was % mortality = 14 + 22 * (age >76) + 9 * (creatinine >1.5) + 20 * (bp <70) Using this model, patients can be stratified into different groups, each with a specific estimated risk of 30-day mortality ranging from a low of 14% to a high of 65%. In the endovascular era where both open and endovascular treatment are offered for the treatment of rAAA three variables, easily obtained in an emergency setting, accurately predict 30-day mortality for patients operated on for rAAA. This simple risk prediction tool could be used as a point of care decision aid to help the clinician in counseling patients and their families on treatment of those presenting with rAAA.


Beyond a motor disorder: a prospective evaluation of sleep quality in cervical dystonia.
Little is known about sleep disturbances in cervical dystonia (CD), particularly the relationship to motoric symptoms. It is critical to clarify these points given the impact on quality of life. Primary CD patients receiving botulinum toxin (BoNT) injections and age- and gender- matched healthy controls were included. In both groups, sleep quality and daytime sleepiness were assessed. In CD, these assessments were repeated following BoNT injections. CD severity, mood symptoms, and health impact of CD were also assessed. 54 CD patients and 55 controls were included. Impaired sleep quality was more frequent in CD compared to controls (t = 4.82, p < 0.0005), even when controlling for the effects of depression, anxiety, and benzodiazepineuse (F = 5.62, p = 0.020). Excessive daytime sleepiness was not significantly different between groups (t = 1.67, p = 0.1). 48 patients received BoNT and returned for follow-up. There was no improvement in sleep quality (t = 0.834, p = 0.41) or daytime somnolence (t = 1.77, p = 0.083) despite improvement in CD severity (t = 4.77, p < 0.0005) with BoNT. There was a small improvement in health impact (t = 2.10, p = 0.04). Sleep quality was more impaired in CD patients, compared to healthy subjects, and did not improve following BoNT treatment, despite a robust improvement in CD severity. This dichotomy suggests that sleep aberrations in CD require separate focus for effective treatment and cannot be viewed as secondary complications of the motor elements of this condition.


Digital chest radiography with photostimulable storage phosphors: signal-to-noise ratio as a function of kilovoltage with matched exposure risk.
A photostimulable storage phosphor (PSP) digital radiography system was evaluated regarding the signal-to-noise ratio (S/N) on chest images acquired at differing peak kilovoltage settings but with matched risk from radiation exposure. Images of two chest phantoms were acquired by using bedside (portable) imaging equipment at tube voltages ranging from 60 to 120 kV. Exposure factors were set so that the effective dose equivalent, a risk estimator weighted for various organs, was approximately equal in all exposures. The S/N in the lung-equivalent regions was found to be slightly higher (maximum, 15%) in the low-energy exposures, while the S/N values in the mediastinum- and subdiaphragm-equivalent regions were approximately equal at all kilovoltage settings. The absence of a high sensitivity of S/N to kilovoltage in risk-matched PSP images should enable institutions to select x-ray beam quality on the basis of other imaging criteria.


First Report of Plasmopara obducens on Impatiens walleriana in Serbia.
In May 2010, Impatiens walleriana plants with single or double flowers that were showing symptoms resembling those of downy mildew were collected in a greenhouse in the vicinity of Mionica, Kolubara District, Serbia. Diseased plants were severely stunted, with mild inconspicuous mottling and yellowing on the upper surface of the leaves. The lower surface of the affected leaves was completely covered with distinctive thick, white fungal-like growth. Symptomatic leaves wilted very quickly and premature leaf fall was common, leaving plants with only a few of the youngest leaves and no or few and poorly developed flowers. Disease incidence was extremely high, approaching 100%, and wilting and collapse of affected plants was very rapid, resulting in losses of more than 90%. White downy growth developing on leaf undersurfaces consisted of hyaline, thin-walled sporangiophores with monopodial branching and numerous, ovoid and hyaline sporangia. Apical branchlets of sporangiophores were at right angles to the main axis, with no apical thickening. Downy mildew of impatiens can be caused by Plasmopara obducens or the less known Bremiella sphaerosperma. The two can be differentiated on the basis of symptomatology and morphology of sporangiophores (1). The absence of well-defined spots on the infected impatiens leaves and straight sporangiophores indicated that the pathogen was P. obducens, which was further supported by molecular identification. Total DNA was extracted directly from plant tissue with a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) following the manufacturer's instructions, and the 5'-end of the nuclear DNA coding for the large ribosomal subunit (LSU rDNA) was amplified by PCR using primers NL1 and NL4 (3). Each PCR amplification yielded two bands estimated at 800 and 650 bp, respectively. A representative isolate, 28-10, was sequenced and 727 bp of the larger band (GenBank Accession No. HQ246451) were found to be identical with P. obducens isolate (AY587558) from the United Kingdom. The sequence was almost identical with those of three P. obducens isolates deposited in NCBI GenBank: EF196869 and AY035522 differed from it by one base pair and FJ787308 by two base pairs. The sequence (HQ223336) of the smaller band was identical to that of three Impatiens accessions (AY727936, AF479154, and AY056515). Pathogenicity tests included inoculation of young I. walleriana plants by spraying with a sporangial suspension. The inoculated plants were kept in experimental chambers at 20°C and 80 to 90% relative humidity, and downy mildew symptoms were observed after 13 to 15 days. To our knowledge, this is the first report of downy mildew of I. walleriana caused by P. obducens in Serbia. So far the presence of P. obducens was recorded in Bulgaria, the Czech Republic, Denmark, Finland, Lithuania, Romania, Russia, the United Kingdom (2), and recently in Norway (4). Thorough inspection would be needed to determine the distribution and incidence of P. obducens on impatiens in Serbia both indoors and outdoors. Impatiens is one of the most popular ornamentals in Serbia and intensive and increasing production may be seriously endangered by the presence of P. obducens. References: (1) O. Constantinescu. Mycologia 83:473, 1991. (2) C. R. Lane et al. Plant Pathol. 54:243, 2005. (3) W. Maier et al. Can. J. Bot. 81:12, 2003. (4) B. Toppe et al. New Dis. Rep. 20:33, 2010.


Craniopagus parasiticus: a case illustrating its relationship to craniopagus conjoined twinning.
A case of craniopagus parasiticus is described in which the parasitic twin is more fully developed anatomically than in any of the previous reports. Somatic and placental vascular anastomoses between the twins and hypoplasia of the umbilical cord of the parasite were also observed. These findings support the hypothesis that craniopagus parasiticus results from compromise of the blood supply to one of a pair of craniopagus conjoined twins.


Characterization of an incompletely assembled major histocompatibility class I molecule (H-2Kb) associated with unusually long peptides: implications for antigen processing and presentation.
We have identified two forms of a major histocompatibility complex (MHC) class I molecule, H-2Kb, distinguishable by specific antibodies through a study of a genetically engineered mouse cell line that overexpresses these molecules. One form, a complex associated with beta 2-microglobulin (native, beta 2m+ class I), is detectable by conformation-dependent antibodies. The other form, which remains after preclearing cell lysates of native class I, is only poorly, if at all, associated with beta 2-microglobulin (beta 2m- class I) and is detectable by an antiserum against the cytoplasmic tail region of H-2K molecules. Both forms are also present in normal cell lines. The affinity-purified native class I molecules bind short peptides (8 or 9 residues) and assemble tightly with beta 2-microglobulin. In striking contrast, the beta 2m- class I molecules bind peptides that are longer (> 15 residues) than those bound to native class I molecules. This finding is consistent with the recent evidence that peptides longer than 8-10 amino acid residues are transported into the endoplasmic reticulum and suggests the possibility of a control step for peptide presentation by MHC in which the incompletely processed peptides bind to the heavy chain and a selected fraction undergoes final processing and presentation on the cell surface.


Genetic and molecular characterization of the optomotor-blind gene locus in Drosophila melanogaster.
The Drosophila gene optomotor-blind (omb) is involved in the development of a set of giant neurons in the optic lobes and possibly other structures in the imaginal brain. Adult flies have discrete defects in optomotor behavior. The gene has previously been mapped in chromomeres 4C5-6, together with three other genes, bifid, Quadroon and lacqueredgls. We have localized the gene in a genomic walk of 340 kb of DNA. By mapping seven chromosome breakpoints with omb phenotype we determined its minimum size to about 80 kb. From this region more than 20 RNAs of different size and temporal expression pattern are transcribed. Three of them (T3, T7 and T7') stem from primary transcripts of 40-80 kb in length. In its distal part the omb gene overlaps in at least 19 kb with four other complementation units, bifid, l(1)bifid, Quadroon and lacqueredgls. The three nonlethals affect the external appearance of the fly and seem to be unrelated to brain development.


Using the EM algorithm to train neural networks: misconceptions and a new algorithm for multiclass classification.
The expectation-maximization (EM) algorithm has been of considerable interest in recent years as the basis for various algorithms in application areas of neural networks such as pattern recognition. However, there exists some misconceptions concerning its application to neural networks. In this paper, we clarify these misconceptions and consider how the EM algorithm can be adopted to train multilayer perceptron (MLP) and mixture of experts (ME) networks in applications to multiclass classification. We identify some situations where the application of the EM algorithm to train MLP networks may be of limited value and discuss some ways of handling the difficulties. For ME networks, it is reported in the literature that networks trained by the EM algorithm using iteratively reweighted least squares (IRLS) algorithm in the inner loop of the M-step, often performed poorly in multiclass classification. However, we found that the convergence of the IRLS algorithm is stable and that the log likelihood is monotonic increasing when a learning rate smaller than one is adopted. Also, we propose the use of an expectation-conditional maximization (ECM) algorithm to train ME networks. Its performance is demonstrated to be superior to the IRLS algorithm on some simulated and real data sets.


Improving intercropping: a synthesis of research in agronomy, plant physiology and ecology.
Intercropping is a farming practice involving two or more crop species, or genotypes, growing together and coexisting for a time. On the fringes of modern intensive agriculture, intercropping is important in many subsistence or low-input/resource-limited agricultural systems. By allowing genuine yield gains without increased inputs, or greater stability of yield with decreased inputs, intercropping could be one route to delivering 'sustainable intensification'. We discuss how recent knowledge from agronomy, plant physiology and ecology can be combined with the aim of improving intercropping systems. Recent advances in agronomy and plant physiology include better understanding of the mechanisms of interactions between crop genotypes and species – for example, enhanced resource availability through niche complementarity. Ecological advances include better understanding of the context-dependency of interactions, the mechanisms behind disease and pest avoidance, the links between above- and below-ground systems, and the role of microtopographic variation in coexistence. This improved understanding can guide approaches for improving intercropping systems, including breeding crops for intercropping. Although such advances can help to improve intercropping systems, we suggest that other topics also need addressing. These include better assessment of the wider benefits of intercropping in terms of multiple ecosystem services, collaboration with agricultural engineering, and more effective interdisciplinary research.


A global comparative evaluation of commercial immunochromatographic rapid diagnostic tests for visceral leishmaniasis.
Poor access to diagnosis stymies control of visceral leishmaniasis (VL). Antibody-detecting rapid diagnostic tests (RDTs) can be performed in peripheral health settings. However, there are many brands available and published reports of variable accuracy. Commercial VL RDTs containing bound rK39 or rKE16 antigen were evaluated using archived human sera from confirmed VL cases (n = 750) and endemic non-VL controls (n = 754) in the Indian subcontinent (ISC), Brazil, and East Africa to assess sensitivity and specificity with 95% confidence intervals. A subset of RDTs were also evaluated after 60 days' heat incubation (37°C, 45°C). Interlot and interobserver variability was assessed. All test brands performed well against ISC panels (sensitivity range, 92.8%-100%; specificity range, 96%-100%); however, sensitivity was lower against Brazil and East African panels (61.5%-91% and 36.8%-87.2%, respectively). Specificity was consistently > 95% in Brazil and ranged between 90.8% and 98% in East Africa. Performance of some products was adversely affected by high temperatures. Agreement between lots and readers was good to excellent (κ > 0.73-0.99). Diagnostic accuracy of VL RDTs varies between the major endemic regions. Many tests performed well and showed good heat stability in the ISC; however, reduced sensitivity against Brazilian and East African panels suggests that in these regions, used alone, several RDTs are inadequate for excluding a VL diagnosis. More research is needed to assess ease of use and to compare performance using whole blood instead of serum and in patients coinfected with human immunodeficiency virus.


Prediction of the binding site of 1-benzyl-4-[(5,6-dimethoxy-1-indanon-2-yl)methyl]piperidine in acetylcholinesterase by docking studies with the SYSDOC program.
In the preceding paper we reported on a docking study with the SYSDOC program for predicting the binding sites of huperzine A in acetylcholinesterase (AChE) [Pang, Y.-P. and Kozikowski, A.P., J. Comput.-Aided Mol. Design, 8 (1994) 669]. Here we present a prediction of the binding sites of 1-benzyl-4-[(5,6-dimethoxy-1-indanon-2-yl)methyl]piperidine (E2020) in AChE by the same method. E2020 is one of the most potent and selective reversible inhibitors of AChE, and this molecule has puzzled researchers, partly due to its flexible structure, in understanding how it binds to AChE. Based on the results of docking 1320 different conformers of E2020 into 69 different conformers of AChE and on the pharmacological data reported for E2020 and its analogs, we predict that both the R- and the S-isomer of E2020 span the whole binding cavity of AChE, with the ammonium group interacting mainly with Trp84, Phe330 and Asp72, the phenyl group interacting mainly with Trp84 and Phe330, and the indanone moiety interacting mainly with Tyr70 and Trp279. The topography of the calculated E2020 binding sites provides insights into understanding the high potency of E2020 in the inhibition of AChE and provides hints as to possible structural modifications for identifying improved AChE inhibitors as potential therapeutics for the palliative treatment of Alzheimer's disease.


Transcription factor profiling in individual hematopoietic progenitors by digital RT-PCR.
We report here a systematic, quantitative population analysis of transcription factor expression within developmental progenitors, made possible by a microfluidic chip-based "digital RT-PCR" assay that can count template molecules in cDNA samples prepared from single cells. In a survey encompassing five classes of early hematopoietic precursor, we found markedly heterogeneous expression of the transcription factor PU.1 in hematopoietic stem cells and divergent patterns of PU.1 expression within flk2- and flk2+ common myeloid progenitors. The survey also revealed significant differences in the level of the housekeeping transcript GAPDH across the surveyed populations, which demonstrates caveats of normalizing expression data to endogenous controls and underscores the need to put gene measurement on an absolute, copy-per-cell basis.


The neuropsychology of sex offenders: a meta-analysis.
Typically, neuropsychological studies of sex offenders have grouped together different types of individuals and different types of measures. This is why results have tended to be nonspecific and divergent across studies. Against this background, the authors undertook a review of the literature regarding the neuropsychology of sex offenders, taking into account subgroups based on criminological theories. They also conducted a meta-analysis of the data to demonstrate the cognitive heterogeneity of sex offenders statistically. Their main objective was to test the hypothesis to the effect that the neuropsychological deficits of sex offenders are not broad and generalized compared with specific subgroups of participants based on specific measures. In all, 23 neuropsychological studies reporting data on 1,756 participants were taken into consideration. As expected, a highly significant, broad, and heterogeneous overall effect size was found. Taking subgroups of participants and specific cognitive measures into account significantly improved homogeneity. Sex offenders against children tended to obtain lower scores than did sex offenders against adults on higher order executive functions, whereas sex offenders against adults tended to obtain results similar to those of non-sex offenders, with lower scores in verbal fluency and inhibition. However, it is concluded that neuropsychological data on sex offenders are still too scarce to confirm these trends or to test more precise hypotheses. For greater clinical relevance, future neuropsychological studies should consider specific subgroups of participants and measures to verify the presence of different cognitive profiles.


An orthotopic glioblastoma animal model suitable for high-throughput screenings.
Glioblastoma (GBM) is an aggressive form of brain cancer with poor prognosis. Although murine animal models have given valuable insights into the GBM disease biology, they cannot be used in high-throughput screens to identify and profile novel therapies. The only vertebrate model suitable for large-scale screens, the zebrafish, has proven to faithfully recapitulate biology and pathology of human malignancies, and clinically relevant orthotopic zebrafish models have been developed. However, currently available GBM orthotopic zebrafish models do not support high-throughput drug discovery screens. We transplanted both GBM cell lines as well as patient-derived material into zebrafish blastulas. We followed the behavior of the transplants with time-lapse microscopy and real-time in vivo light-sheet microscopy. We found that GBM material transplanted into zebrafish blastomeres robustly migrated into the developing nervous system, establishing an orthotopic intracranial tumor already 24 hours after transplantation. Detailed analysis revealed that our model faithfully recapitulates the human disease. We have developed a robust, fast, and automatable transplantation assay to establish orthotopic GBM tumors in zebrafish. In contrast to currently available orthotopic zebrafish models, our approach does not require technically challenging intracranial transplantation of single embryos. Our improved zebrafish model enables transplantation of thousands of embryos per hour, thus providing an orthotopic vertebrate GBM model for direct application in drug discovery screens.


Functional cross-talk between phosphorylation and disease-causing mutations in the cardiac sodium channel Nav1.5.
The voltage-gated sodium channel Nav1.5 initiates the cardiac action potential. Alterations of its activation and inactivation properties due to mutations can cause severe, life-threatening arrhythmias. Yet despite intensive research efforts, many functional aspects of this cardiac channel remain poorly understood. For instance, Nav1.5 undergoes extensive posttranslational modification in vivo, but the functional significance of these modifications is largely unexplored, especially under pathological conditions. This is because most conventional approaches are unable to insert metabolically stable posttranslational modification mimics, thus preventing a precise elucidation of the contribution by these modifications to channel function. Here, we overcome this limitation by using protein semisynthesis of Nav1.5 in live cells and carry out complementary molecular dynamics simulations. We introduce metabolically stable phosphorylation mimics on both wild-type (WT) and two pathogenic long-QT mutant channel backgrounds and decipher functional and pharmacological effects with unique precision. We elucidate the mechanism by which phosphorylation of Y1495 impairs steady-state inactivation in WT Nav1.5. Surprisingly, we find that while the Q1476R patient mutation does not affect inactivation on its own, it enhances the impairment of steady-state inactivation caused by phosphorylation of Y1495 through enhanced unbinding of the inactivation particle. We also show that both phosphorylation and patient mutations can impact Nav1.5 sensitivity toward the clinically used antiarrhythmic drugs quinidine and ranolazine, but not flecainide. The data highlight that functional effects of Nav1.5 phosphorylation can be dramatically amplified by patient mutations. Our work is thus likely to have implications for the interpretation of mutational phenotypes and the design of future drug regimens.


An evaluation of different protocols for measuring the force-velocity relationship of the human quadriceps muscles.
A modified Cybex II isokinetic dynamometer was used to evaluate the problems associated with measuring the concentric force-velocity characteristics of human knee extensor muscles. Three contraction protocols were investigated, simple voluntary contractions (VC); releases from maximal voluntary isometric contractions (VR) and releases from isometric femoral nerve stimulated contractions (FNR). Percutaneous stimulation of the quadriceps was unsuitable for dynamic contractions as the proportion of the muscle activated varied with the angle of knee flexion. Isometric length-tension relationships and isokinetic contractions at seven angular velocities between 0.5 and 5.2 rad.s-1 were recorded in five subjects. During isometric and slow dynamic contractions the voluntary forces were often greater than those obtained by femoral nerve stimulation, probably due to subjects stretching the rectus femoris during voluntary manoeuvres. It was found that the VC protocol produced acceptable isokinetic force recordings only at velocities below 3.1 rad.s-1 in most subjects whilst VR contractions resulted in unexpectedly low forces at velocities below 1.57 rad.s-1. Of the three techniques employed, FNR, although uncomfortable for subjects, provided the most accurate and reliable method of measuring force-velocity characteristics of knee extensor muscles. FNR contractions produced a force-velocity curve which showed a smooth decline in force with increasing velocity up to 5.2 rad.s-1. VC contractions appear to be an acceptable alternative for testing the muscles provided the angular velocity is less than 3.1 rad.s-1 and the subjects can be prevented from stretching the rectus femoris during the movement.


Couple and family involvement in adult mental health treatment: a systematic review.
We reviewed randomized controlled trials conducted in the United States from January, 1996 through December, 2011 that examined family interventions for adult mental health conditions. We identified 51 articles (39 trials) evaluating 21 different family interventions. Findings for behavioral couple or family therapy (BCT/BFT) and community reinforcement and training (CRAFT) for substance use disorders were each pooled separately for examination in meta-analyses. Findings suggest BCT/BFT reduced substance use (small-to-moderate effects) and improved relationship adjustment (large effects) compared to individually-oriented treatments. CRAFT increased treatment initiation three-fold but did not improve substance use or family functioning over alternative family interventions. Family focused therapy for bipolar disorder improved symptoms over less intensive treatments with mixed findings when compared to equally intensive treatments. For both bipolar disorder and schizophrenia spectrum disorders, the few trials meeting our search criteria and heterogeneity among trials precluded generating broader conclusions regarding which family interventions are most effective for US populations. Overall, trials were limited in their methodological quality, and many interventions were evaluated in one trial. Future research is needed to replicate findings for these single trials, examine relationship distress as a moderator of outcome, and examine BCT/BFT among dual substance using couples and outside the research group frequently represented.


Male gametic cell-specific gene expression in flowering plants.
The role of the male gamete-the sperm cell-in the process of fertilization is to recognize, adhere to, and fuse with the female gamete. These highly specialized functions are expected to be controlled by activation of a unique set of genes. However, male gametic cells traditionally have been regarded as transcriptionally quiescent because of highly condensed chromatin and a very reduced amount of cytoplasm. Here, we provide evidence for male gamete-specific gene expression in flowering plants. We identified and characterized a gene, LGC1, which was shown to be expressed exclusively in the male gametic cells. The gene product of LGC1 was localized at the surface of male gametic cells, suggesting a possible role in sperm-egg interactions. These findings represent an important step toward defining the molecular mechanisms of male gamete development and the cellular processes involved in fertilization of flowering plants.


Soft-molecular imprinted electrospun scaffolds to mimic specific biological tissues.
The fabrication of bioactive scaffolds able to mimic the in vivo cellular microenvironment is a challenge for regenerative medicine. The creation of sites for the selective binding of specific endogenous proteins represents an attractive strategy to fabricate scaffolds able to elicit specific cell response. Here, electrospinning (ESP) and soft-molecular imprinting (soft-MI) techniques were combined to fabricate a soft-molecular imprinted electrospun bioactive scaffold (SMIES) for tissue regeneration. Scaffolds functionalized using different proteins and growth factors (GFs) arranged onto the surface were designed, fabricated and validated with different polyesters, demonstrating the versatility of the developed approach. The scaffolds bound selectively each of the different proteins used, indicating that the soft-MI method allowed fabricating high affinity binding sites on ESP fibers compared to non-imprinted ones. The imprinting of ESP fibers with several GFs resulted in a significant effect on cell behavior. FGF-2 imprinted SMIES promoted cell proliferation and metabolic activity. BMP-2 and TGF-β3 imprinted SMIES promoted cellular differentiation. These scaffolds hold the potential to be used in a cell-free approach to steer endogenous tissue regeneration in several regenerative medicine applications.


Ligand-binding affinity of alternative conformers of human β2 -adrenergic receptor in the presence of intracellular loop 3 (ICL3) and their potential use in virtual screening studies.
This study investigates the structural distinctiveness of orthosteric ligand-binding sites of several human β2 adrenergic receptor (β2 -AR) conformations that have been obtained from a set of independent molecular dynamics (MD) simulations in the presence of intracellular loop 3 (ICL3). A docking protocol was established in order to classify each receptor conformation via its binding affinity to selected ligands with known efficacy. This work's main goal was to reveal many subtle features of the ligand-binding site, presenting alternative conformations, which might be considered as either active- or inactive-like but mostly specific for that ligand. Agonists, inverse agonists, and antagonists were docked to each MD conformer with distinct binding pockets, using different docking tools and scoring functions. Mostly favored receptor conformation persistently observed in all docking/scoring evaluations was classified as active or inactive based on the type of ligand's biological effect. Classified MD conformers were further tested for their ability to discriminate agonists from inverse agonists/antagonists, and several conformers were proposed as important targets to be used in virtual screening experiments that were often limited to a single X-ray structure.


Natural resistance to ascorbic acid induced oxidative stress is mainly mediated by catalase activity in human cancer cells and catalase-silencing sensitizes to oxidative stress.
Ascorbic acid demonstrates a cytotoxic effect by generating hydrogen peroxide, a reactive oxygen species (ROS) involved in oxidative cell stress. A panel of eleven human cancer cell lines, glioblastoma and carcinoma, were exposed to serial dilutions of ascorbic acid (5-100 mmol/L). The purpose of this study was to analyse the impact of catalase, an important hydrogen peroxide-detoxifying enzyme, on the resistance of cancer cells to ascorbic acid mediated oxidative stress. Effective concentration (EC(50)) values, which indicate the concentration of ascorbic acid that reduced the number of viable cells by 50%, were detected with the crystal violet assay. The level of intracellular catalase protein and enzyme activity was determined. Expression of catalase was silenced by catalase-specific short hairpin RNA (sh-RNA) in BT-20 breast carcinoma cells. Oxidative cell stress induced apoptosis was measured by a caspase luminescent assay. The tested human cancer cell lines demonstrated obvious differences in their resistance to ascorbic acid mediated oxidative cell stress. Forty-five percent of the cell lines had an EC(50) > 20 mmol/L and fifty-five percent had an EC(50) < 20 mmol/L. With an EC(50) of 2.6-5.5 mmol/L, glioblastoma cells were the most susceptible cancer cell lines analysed in this study. A correlation between catalase activity and the susceptibility to ascorbic acid was observed. To study the possible protective role of catalase on the resistance of cancer cells to oxidative cell stress, the expression of catalase in the breast carcinoma cell line BT-20, which cells were highly resistant to the exposure to ascorbic acid (EC(50): 94,9 mmol/L), was silenced with specific sh-RNA. The effect was that catalase-silenced BT-20 cells (BT-20 KD-CAT) became more susceptible to high concentrations of ascorbic acid (50 and 100 mmol/L). Fifty-five percent of the human cancer cell lines tested were unable to protect themselves against oxidative stress mediated by ascorbic acid induced hydrogen peroxide production. The antioxidative enzyme catalase is important to protect cancer cells against cytotoxic hydrogen peroxide. Silenced catalase expression increased the susceptibility of the formerly resistant cancer cell line BT-20 to oxidative stress.


Inflammatory papillary hyperplasia: review of literature and case report involving a 10-year-old child.
Inflammatory papillary hyperplasia is a benign lesion of the palate seen most often in patients with a history of ill-fitting dentures or poor oral hygiene. The specific cause is unknown. Inflammatory papillary hyperplasia can occur at any age. However, it is most often seen in patients in the third to fifth decades. It occurs more frequently in males and whites. The best treatment is surgical removal. The prognosis is excellent, once the lesion is removed. The patient presented in this case report is a 10-year-old black girl without a history of a dental prosthesis. It is conjectured that poor oral hygiene and a habit of mouth breathing contributed to the occurrence of inflammatory papillary hyperplasia in this patient. The lesion was surgically removed, and the patient was followed up for a period of 18 months without recurrence of the lesion.


Crebinostat: a novel cognitive enhancer that inhibits histone deacetylase activity and modulates chromatin-mediated neuroplasticity.
Long-term memory formation is known to be critically dependent upon de novo gene expression in the brain. As a consequence, pharmacological enhancement of the transcriptional processes mediating long-term memory formation provides a potential therapeutic strategy for cognitive disorders involving aberrant neuroplasticity. Here we focus on the identification and characterization of small molecule inhibitors of histone deacetylases (HDACs) as enhancers of CREB (cAMP response element-binding protein)-regulated transcription and modulators of chromatin-mediated neuroplasticity. Using a CREB reporter gene cell line, we screened a library of small molecules structurally related to known HDAC inhibitors leading to the identification of a probe we termed crebinostat that produced robust activation of CREB-mediated transcription. Further characterization of crebinostat revealed its potent inhibition of the deacetylase activity of recombinant class I HDACs 1, 2, 3, and class IIb HDAC6, with weaker inhibition of the class I HDAC8 and no significant inhibition of the class IIa HDACs 4, 5, 7, and 9. In cultured mouse primary neurons, crebinostat potently induced acetylation of both histone H3 and histone H4 as well as enhanced the expression of the CREB target gene Egr1 (early growth response 1). Using a hippocampus-dependent, contextual fear conditioning paradigm, mice systemically administered crebinostat for a ten day time period exhibited enhanced memory. To gain insight into the molecular mechanisms of memory enhancement by HDAC inhibitors, whole genome transcriptome profiling of cultured mouse primary neurons treated with crebinostat, combined with bioinformatic analyses of CREB-target genes, was performed revealing a highly connected protein-protein interaction network reflecting modules of genes important to synaptic structure and plasticity. Consistent with these findings, crebinostat treatment increased the density of synapsin-1 punctae along dendrites in cultured neurons. Finally, crebinostat treatment of cultured mouse primary neurons was found to upregulate Bdnf (brain-derived neurotrophic factor) and Grn (granulin) and downregulate Mapt (tau) gene expression-genes implicated in aging-related cognitive decline and cognitive disorders. Taken together, these results demonstrate that crebinostat provides a novel probe to modulate chromatin-mediated neuroplasticity and further suggests that pharmacological optimization of selective of HDAC inhibitors may provide an effective therapeutic approach for human cognitive disorders. This article is part of a Special Issue entitled 'Cognitive Enhancers'.


The relationship of perceived neighborhood social climate to walking in Hispanic older adults: a longitudinal, cross-lagged panel analysis.
This study examines possible bidirectional relationships between neighborhood climate (i.e., perceived neighborhood social environment) and walking behavior across a 12-month period in older Hispanics. A population-based sample of 217 community-dwelling older Hispanics in Miami, Florida, completed measures of perceived neighborhood climate and neighborhood walking, at two assessment time points (12 months apart). Structural equation modeling analyses revealed that neighborhood climate predicted subsequent walking 12 months later, such that more positive perceptions of neighborhood climate predicted more walking. Follow-up analyses revealed that older adults who resided in the top half of neighborhoods based on perceived neighborhood climate scores at initial assessment were 2.57 times as likely to have walked at least one block in the last week at follow-up, relative to older adults residing in neighborhoods whose climate was in the lower half. Perceptions of a more positive neighborhood social environment may promote walking in urban, older Hispanics.


Magnetic nanoparticles for ultrafast mechanical control of inner ear hair cells.
We introduce cubic magnetic nanoparticles as an effective tool for precise and ultrafast control of mechanosensitive cells. The temporal resolution of our system is ∼1000 times faster than previously used magnetic switches and is comparable to the current state-of-the-art optogenetic tools. The use of a magnetism-gated switch reported here can address the key challenges of studying mechanotransduction in biological systems. The cube-shaped magnetic nanoparticles are designed to bind to components of cellular membranes and can be controlled with an electromagnet to exert pico-Newtons of mechanical force on the cells. The cubic nanoparticles can thus be used for noncontact mechanical control of the position of the stereocilia of an inner ear hair cell, yielding displacements of tens of nanometers, with sub-millisecond temporal resolution. We also prove that such mechanical stimulus leads to the influx of ions into the hair cell. Our study demonstrates that a magnetic switch can yield ultrafast temporal resolution, and has capabilities for remote manipulation and biological specificity, and that such magnetic system can be used for the study of mechanotransduction processes of a wide range of sensory systems.


Sympathetic neuron-associated macrophages contribute to obesity by importing and metabolizing norepinephrine.
The cellular mechanism(s) linking macrophages to norepinephrine (NE)-mediated regulation of thermogenesis have been a topic of debate. Here we identify sympathetic neuron-associated macrophages (SAMs) as a population of cells that mediate clearance of NE via expression of solute carrier family 6 member 2 (SLC6A2), an NE transporter, and monoamine oxidase A (MAOA), a degradation enzyme. Optogenetic activation of the sympathetic nervous system (SNS) upregulates NE uptake by SAMs and shifts the SAM profile to a more proinflammatory state. NE uptake by SAMs is prevented by genetic deletion of Slc6a2 or inhibition of the encoded transporter. We also observed an increased proportion of SAMs in the SNS of two mouse models of obesity. Genetic ablation of Slc6a2 in SAMs increases brown adipose tissue (BAT) content, causes browning of white fat, increases thermogenesis, and leads to substantial and sustained weight loss in obese mice. We further show that this pathway is conserved, as human sympathetic ganglia also contain SAMs expressing the analogous molecular machinery for NE clearance, which thus constitutes a potential target for obesity treatment.


Health professionals' experiences with the implementation of a digital medication dispenser in home care services - a qualitative study.
Implementing digital technology in home care services challenges care arrangements built on face-to-face encounters. Digital welfare technology has been suggested as a solution to increasing demands on health care services from an ageing population. Medication delivery is a major task for home care services, and digital medication devices could lessen the need for resources. But technology has scripts based on how designers picture its use, and these might not fit with users' needs and practices. New technology must go through processes of domestication among its users. In the present study, we investigate how health professionals experienced the implementation of a digital medication dispenser into home care services in Norway. This was a qualitative interview study with 26 health professionals from home care services in five municipalities. All five municipalities had implemented a digital medication dispenser in home care services. Prior to the introduction of the dispenser, medication practices had been based on home visits. The safety of medication practices was the main concern of health professionals who had to negotiate the technological script in order to make it work in a new care arrangement. Rationalities of effectiveness collided with rationalities of care, symbolized by warm hands. Professionals who had been used to working independently became dependent on technical support. Being unfamiliar with the new medication arrangement led to resistance towards the digital dispenser, but more direct experiences changed the focus from technology to new care arrangements. Negotiating practical and organizational arrangements led health professionals to trust the digital medication dispenser to contribute to safe and good care for service users. Implementing digital technology in home care services must be informed by previous practices in the field, especially when it concerns safety for patients. Through processes of domestication, health professionals negotiate technological scripts to make them fit professional ideals and practices. Policymakers and managers must address questions of care arrangements and individualized adaptions to patients' needs in order to receive support from health professionals when implementing digital technology in home care services.


Rat intersubjective decisions are encoded by frequency-specific oscillatory contexts.
It is unknown how the brain coordinates decisions to withstand personal costs in order to prevent other individuals' distress. Here we test whether local field potential (LFP) oscillations between brain regions create "neural contexts" that select specific brain functions and encode the outcomes of these types of intersubjective decisions. Rats participated in an "Intersubjective Avoidance Test" (IAT) that tested rats' willingness to enter an innately aversive chamber to prevent another rat from getting shocked. c-Fos immunoreactivity was used to screen for brain regions involved in IAT performance. Multi-site local field potential (LFP) recordings were collected simultaneously and bilaterally from five brain regions implicated in the c-Fos studies while rats made decisions in the IAT. Local field potential recordings were analyzed using an elastic net penalized regression framework. Rats voluntarily entered an innately aversive chamber to prevent another rat from getting shocked, and c-Fos immunoreactivity in brain regions known to be involved in human empathy-including the anterior cingulate, insula, orbital frontal cortex, and amygdala-correlated with the magnitude of "intersubjective avoidance" each rat displayed. Local field potential recordings revealed that optimal accounts of rats' performance in the task require specific frequencies of LFP oscillations between brain regions in addition to specific frequencies of LFP oscillations within brain regions. Alpha and low gamma coherence between spatially distributed brain regions predicts more intersubjective avoidance, while theta and high gamma coherence between a separate subset of brain regions predicts less intersubjective avoidance. Phase relationship analyses indicated that choice-relevant coherence in the alpha range reflects information passed from the amygdala to cortical structures, while coherence in the theta range reflects information passed in the reverse direction. These results indicate that the frequency-specific "neural context" surrounding brain regions involved in social cognition encodes outcomes of decisions that affect others, above and beyond signals from any set of brain regions in isolation.


Hepatic steatosis and cardiovascular disease outcomes: An analysis of the Framingham Heart Study.
Non-alcoholic fatty liver disease (NAFLD) is highly prevalent and is associated with development of metabolic disease including atherosclerotic cardiovascular disease (CVD). Our aim is to examine the association of hepatic steatosis with prevalent clinical and subclinical CVD outcomes in a large community-based sample, the Framingham Heart Study. Hepatic steatosis was measured in 3529 participants using multidetector computed tomography scanning. Multivariable logistic regression was used to determine whether hepatic steatosis is associated with prevalent CVD adjusted for covariates. We also tested whether associations were independent of other metabolic diseases/traits. The primary clinical outcome was composite prevalent clinical CVD defined by prior non-fatal myocardial infarction, stroke, transient ischemic attack, heart failure, or peripheral arterial disease. Subclinical cardiovascular outcomes were coronary artery calcium (CAC) and abdominal artery calcium (AAC). 3014 participants were included (50.5% women). There was a non-significant association of hepatic steatosis with clinical CVD (OR 1.14 [p=0.07]). Hepatic steatosis was associated with both CAC and AAC (OR 1.20 [p<0.001] and OR 1.16 [p<0.001], respectively). Associations persisted for CAC even when controlling for other risk factors/metabolic diseases, but for AAC, the associations became non-significant after adjustment for visceral adipose tissue. The association between hepatic steatosis and AAC was stronger in men than in women (p sex interaction=0.022). There was a significant association of hepatic steatosis with subclinical CVD outcomes independent of many metabolic diseases/traits with a trend towards association between hepatic steatosis and clinical CVD outcomes. The association with AAC was stronger in men than in women.


A microcontroller-based simulation of dural venous sinus injury for neurosurgical training.
OBJECTIVE Surgical simulation has the potential to supplement and enhance traditional resident training. However, the high cost of equipment and limited number of available scenarios have inhibited wider integration of simulation in neurosurgical education. In this study the authors provide initial validation of a novel, low-cost simulation platform that recreates the stress of surgery using a combination of hands-on, model-based, and computer elements. Trainee skill was quantified using multiple time and performance measures. The simulation was initially validated using trainees at the start of their intern year. METHODS The simulation recreates intraoperative superior sagittal sinus injury complicated by air embolism. The simulator model consists of 2 components: a reusable base and a disposable craniotomy pack. The simulator software is flexible and modular to allow adjustments in difficulty or the creation of entirely new clinical scenarios. The reusable simulator base incorporates a powerful microcomputer and multiple sensors and actuators to provide continuous feedback to the software controller, which in turn adjusts both the screen output and physical elements of the model. The disposable craniotomy pack incorporates 3D-printed sections of model skull and brain, as well as artificial dura that incorporates a model sagittal sinus. RESULTS Twelve participants at the 2015 Western Region Society of Neurological Surgeons postgraduate year 1 resident course ("boot camp") provided informed consent and enrolled in a study testing the prototype device. Each trainee was required to successfully create a bilateral parasagittal craniotomy, repair a dural sinus tear, and recognize and correct an air embolus. Participant stress was measured using a heart rate wrist monitor. After participation, each resident completed a 13-question categorical survey. CONCLUSIONS All trainee participants experienced tachycardia during the simulation, although the point in the simulation at which they experienced tachycardia varied. Survey results indicated that participants agreed the simulation was realistic, created stress, and was a useful tool in training neurosurgical residents. This simulator represents a novel, low-cost approach for hands-on training that effectively teaches and tests residents without risk of patient injury.


Inhibition of lung tumorigenesis in A/J mice by N-acetyl-S-(N-2-phenethylthiocarbamoyl)-L-cysteine and myo-inositol, individually and in combination.
Isothiocyanates, their N-acetylcysteine conjugates, and myo-inositol (MI) are inhibitors of lung tumorigenesis in A/J mice. However, chemoprevention by combinations of these compounds in different temporal sequences has not been examined. This is important for developing practical approaches to lung cancer chemoprevention in smokers and ex-smokers. We used a tumor model in which A/J mice are treated with 8 weekly doses of benzo[a]pyrene (B[a]P) plus 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and killed 19 weeks after the final treatment. In Experiment 1, isothiocyanates or their N-acetylcysteine conjugates were added to the diet (1 or 3 micro mol/g) from 1 week before until 1 week after carcinogen treatment. The compounds were 2-phenethyl isothiocyanate (PEITC), 3-phenylpropyl isothiocyanate (PPITC), N-acetyl-S-(N-benzyl-thiocarbamoyl)-L-cysteine (BITC-NAC), N-acetyl-S-(N-2-phenethylthiocarbamoyl)-L-cysteine (PEITC-NAC), and N-acetyl-S-(N-3-phenylpropylthiocarbamoyl)-L-cysteine (PPITC-NAC). Significant reductions in lung tumor multiplicity were observed in mice treated with PEITC, PEITC-NAC, PPITC and PPITC-NAC. PEITC-NAC was chosen for combination studies with MI (Experiment 2). Mice were treated with B[a]P plus NNK without or with PEITC-NAC (3 micro mol/g diet), MI (55.5 micro mol/g diet), or PEITC-NAC plus MI (3 micro mol plus 55.5 micro mol/g diet). Different temporal sequences of dietary additions were investigated: carcinogen treatment phase; post-carcinogen treatment phase; entire experiment; 50% of carcinogen treatment phase until termination; and 75% of carcinogen treatment phase until termination. All treatments reduced lung tumor multiplicity except PEITC-NAC post-carcinogen or from 75% of the carcinogen treatment phase. Reduction of lung tumor multiplicity by PEITC-NAC plus MI was greater than that in the mice treated with the agents alone in all temporal sequences. When all results were combined, PEITC-NAC plus MI was significantly more effective than the agents alone. There was a significant trend for reduction in lung tumor multiplicity with increased duration of treatment by the chemopreventive agents. These results provide a basis for further development of mixtures of PEITC-NAC and MI for chemoprevention of lung cancer.


Evidence-based practice in primary health care.
This study aims to describe primary care professionals' self-reported attitudes towards evidence-based practice (EBP), attention to information sources, perceptions of the barriers to EBP and strategies to improve insight in EBP and patient care. An e-mail invitation with link to an Internet-based survey was sent to Belgian medical doctors (MDs), nurses and paramedics. Under paramedics, we've included emergency medical technicians, firemen and medical volunteers (Red Cross). In general, respondents were supportive towards EBP and agreed that this concept improves patient care, but still, physicians claim that only 50% of their practice is evidence-based and nurses and paramedics spend respectively 59% and 54% of their time to EBP. Doctors depend mostly on clinical guidelines, the Internet and textbooks, while nurses prefer conferences and protocols and paramedics rely on courses and their own judgement. All respondents strongly rely on experimental knowledge gained through interaction with colleagues, although the majority reported that colleagues are often not supportive towards EBP. Lack of time, the overwhelming mass of literature, difficulties with implementation of evidence in to practice are the most common barriers. Nurses show lack of critical appraisal of research results and paramedics have difficulties understanding research and have limited access to computer facilities and their working environment. Communication in group and workshops are very highly valued. Nurses and paramedics are less reluctant towards the opinion of senior colleagues, audits on clinical practice and individual feedback than doctors. EBP generally enjoys a positive attitude at every level of the health care system, but still many obstacles have to be overcome to conquer 'experience-based practice'. The most appropriate method for actual implementation of evidence-based practice at all levels of health care is to provide summaries of evidence, easily understandable protocols and web-based databases accessible from the working environment. Students should not only learn the skills related to EBP, but should be able to integrate knowledge effectively in the clinical setting and routine care. Above all, their supervisors themselves need to evolve from 'experience-based' to evidence-based practice'.


Exploring men's and women's experiences of depression and engagement with health professionals: more similarities than differences? A qualitative interview study.
It is argued that the ways in which women express emotional distress mean that they are more likely to be diagnosed with depression, while men's relative lack of articulacy means their depression is hidden. This may have consequences for communicating with health professionals. The purpose of this analysis was to explore how men and women with depression articulate their emotional distress, and examine whether there are gender differences or similarities in the strategies that respondents found useful when engaging with health professionals. In-depth qualitative interviews with 22 women and 16 men in the UK who identified themselves as having had depression, recruited through general practitioners, psychiatrists and support groups. We found gender similarities and gender differences in our sample. Both men and women found it difficult to recognise and articulate mental health problems and this had consequences for their ability to communicate with health professionals. Key gender differences noted were that men tended to value skills which helped them to talk while women valued listening skills in health professionals, and that men emphasised the importance of getting practical results from talking therapies in their narratives, as opposed to other forms of therapy which they conceptualised as 'just talking'. We also found diversity among women and among men; some respondents valued a close personal relationship with health professionals, while others felt that this personal relationship was a barrier to communication and preferred 'talking to a stranger'. Our findings suggest that there is not a straightforward relationship between gender and engagement with health professionals for people with depression. Health professionals need to be sensitive to patients who have difficulties in expressing emotional distress and critical of gender stereotypes which suggest that women invariably find it easy to express emotional distress and men invariably find it difficult. In addition it is important to recognise that, for a minority of patients, a personal relationship with health professionals can act as a barrier to the disclosure of emotional distress.


Progress and priorities in the health of women and girls: a decade of advances and challenges.
Following the initial wave of federal support to address women's health, there is a need to assess successes and determine the next priorities to advance the health of women. The objective of this study was to systematically collect expert opinion on the major advances in women's health in the past decade and priorities for women's health research and service in the coming decade. We utilized a Delphi method to query the leadership from academic and community Centers of Excellence in Women's Health, as designated by the Department of Health and Human Services. Leaders from 36 of the 48 centers responded to a series of questions about the major advances and critical indicators to evaluate future needs in women's health. We utilized a social ecology model framework to organize the responses to each question. The experts identified increased health education for women and increased empowerment of women across multiple spheres as the major advances positively impacting the health of women. The experts selected the following areas as the most important indicators to measure the status of the health of women in the future: health education and promotion, rates and impact of interpersonal violence against women, and access to healthcare. The major advances and measures of the health of women did not focus on specific changes to individual women in illness management, clinical care, or individual behavioral change. As we move to address health reform, we must be able to recognize and incorporate a broad perspective on public health and policy initiatives critical to the health and wellness of women and girls and, therefore, central to the well-being of the nation.


Imaging profiles of light intensity in the near field: applications to phase-shift photolithography.
We describe a method of imaging the intensity profiles of light in near-field lithographic experiments directly by using a sensitive photoresist. This technique was applied to a detailed study of the irradiance distribution in the optical near field with contact-mode photolithography carried out by use of elastomeric phase masks. The experimental patterns in the photoresist determined by scanning electron microscopy and atomic force microscopy were compared with the corresponding theoretical profiles of intensity calculated by use of a simple scalar analysis; the two correlate well. This comparison makes it possible to improve the theoretical models of irradiance distribution in the near field. Analysis of the images highlights issues in the experimental design, provides a means for the optimization of this technique, and extends its application to the successful fabrication of test structures with linewidths of ~50 nm.