Forskolin inhibits and reverses the effects of brefeldin A on Golgi morphology by a cAMP-independent mechanism.
Brefeldin A (BFA) causes rapid redistribution of Golgi proteins into the ER, leaving no definable Golgi apparatus, and blocks transport of proteins into post-Golgi compartments in the cell. In this study we follow the disassembly of the Golgi apparatus in BFA-treated, living cells labeled with NBD-ceramide and demonstrate that forskolin can both inhibit and reverse this process. Long, tubular processes labeled with NBD-ceramide were observed emerging from Golgi elements and extending out to the cell periphery in cells treated with BFA for 5 min. With longer incubations in BFA, the NBD label was dispersed in a fine reticular pattern characteristic of the ER. Treatment with forskolin inhibited these effects of BFA as well as BFA's earliest morphologic effect on the Golgi apparatus: the redistribution to the cytosol of a 110-kD Golgi peripheral membrane protein. In addition, forskolin could reverse BFA's block in protein secretion. Forskolin inhibition of BFA's effects was dose dependent and reversible. High concentrations of BFA could overcome forskolin's inhibitory effect, suggesting forskolin and BFA interact in a competitive fashion. Remarkably, in cells already exposed to BFA, forskolin could reverse BFA's effects causing the 110-kD Golgi peripheral membrane protein to reassociate with Golgi membrane and juxtanuclear Golgi complexes to reassemble. Neither membrane permeant cAMP analogues nor cAMP phosphodiesterase inhibitors could replicate or enhance forskolin's inhibition of BFA. 1,9-Dideoxyforskolin, which does not activate adenylyl cyclase, was equally as effective as forskolin in antagonizing BFA. A derivative of forskolin, 7-HPP-forskolin, that is less potent than forskolin at binding to adenylyl cyclase, was also equally effective as forskolin in antagonizing BFA. In contrast a similar derivative, 6-HPP-forskolin, that is equipotent with forskolin at binding to adenylyl cyclase, did not inhibit BFA's effects. These results suggest that forskolin acts as a competitive antagonist to BFA, using a cAMP-independent mechanism to prevent and reverse the morphologic effects induced by BFA.


US nativity and dietary acculturation impact the gut microbiome in a diverse US population.
Little is known regarding the impact of immigrant acculturation on the gut microbiome. We characterized differences in the gut microbiome between racially/ethnically diverse US immigrant and US-born groups, and determined the impact of dietary acculturation on the microbiome. Stool samples were collected from 863 US residents, including US-born (315 White, 93 Black, 40 Hispanic) and foreign-born (105 Hispanic, 264 Korean) groups. We determined dietary acculturation from dissimilarities based on food frequency questionnaires, and used 16S rRNA gene sequencing to characterize the microbiome. Gut microbiome composition differed across study groups, with the largest difference between foreign-born Koreans and US-born Whites, and significant differences also observed between foreign-born and US-born Hispanics. Differences in sub-operational taxonomic unit (s-OTU) abundance between foreign-born and US-born groups tended to be distinct from differences between US-born groups. Bacteroides plebeius, a seaweed-degrading bacterium, was strongly enriched in foreign-born Koreans, while Prevotella copri and Bifidobacterium adolescentis were strongly enriched in foreign-born Koreans and Hispanics, compared with US-born Whites. Dietary acculturation in foreign-born participants was associated with specific s-OTUs, resembling abundance in US-born Whites; e.g., a Bacteroides plebeius s-OTU was depleted in highly diet-acculturated Koreans. In summary, we observed that US nativity is a determinant of the gut microbiome in a US resident population. Dietary acculturation may result in loss of native species in immigrants, though further research is necessary to explore whether acculturation-related microbiome alterations have consequences for immigrant health.


Angiotensin delays platelet aggregation after injury of cerebral arterioles.
Endothelium of cerebral surface vessels (pial arterioles and venules) was injured with a light/dye technique in anesthetized mice. This induced platelet aggregation at the site of injury. The onset of aggregation was monitored through a microscope in mice given angiotensin II acetate, 4 micrograms i.v. 30 minutes earlier. Aggregation latency was compared with that in vehicle treated (saline) mice. Angiotensin II caused a highly significant delay in aggregation within the arterioles which was not related to a change in shear rate of blood. Angiotensin II added to platelet rich plasma, failed to influence the aggregation produced by subsequent addition of 0.5 microM ADP or 0.5 mM sodium arachidonate. Angiotensin is a well known stimulator of prostacyclin synthesis or release, and angiotensin has been reported to inhibit platelet aggregation ex vivo by increasing prostacyclin in the effluent superfusing the mass of aggregating platelets. Our data represent the first report of an antiaggregating effect of angiotensin II in vivo in an intact microvascular bed. The data is consonant with the literature describing increased prostacyclin levels following angiotensin II infusion. The antiaggregating effect of angiotensin in cerebral microvessels may help explain a recent observation describing increased survival of gerbils treated with angiotensin following carotid ligation.


Identification of a Staphylococcus aureus extracellular matrix-binding protein with broad specificity.
A staphylococal surface protein capable of binding several extracellular matrix glycoproteins was purified as a result of our attempts to identify a receptor(s) for bone sialoprotein (BSP) on Staphylococcus aureus cells. Proteins from different staphylococcal strains were solubilized in sodium lauryl sulfate, separated by polyacrylamide gel electrophoresis, blotted onto Immobilon P membranes, and probed with 125I-BSP. Several bacterial proteins bound the radiolabeled ligand, and various strains expressed different repertoirs of BSP-binding proteins. Major BSP-binding proteins with apparent M(r)s of 72,000 or 60,000 were present on most strains, and these proteins were further studied. The 72- and 60-kDa proteins were preferentially expressed when bacteria were cultured in Luria broth compared with when they were cultured on tryptic soy broth, and the abundance of the proteins could be correlated to an increased 125I-BSP binding. Both the 72-kDa and the 60-kDa proteins were solubilized by extraction of cells with 1 M LiCl and were purified by cation-exchange chromatography. Amino acid composition analysis of the purified 72-kDa protein indicated a high content of lysine (11.9%) and hydrophobic amino acids (28.0% combined). In Western ligand blotting (immunoblotting) experiments, the 72-kDa protein bound not only BSP but also radiolabeled fibronectin, fibrinogen, vitronectin, thrombospondin, and, to some extent, collagen. Addition of the purified 60-kDa protein to S. aureus cells did not inhibit binding of the different ligands but in some cases resulted in an augmentation of the binding of 125I-ligand. Purified 60-kDa protein could hemagglutinate sheep erythrocytes at a concentration of 61 micrograms/ml. The agglutination reaction was inhibited by high concentrations of fucose, mannose, or melibiose. These data suggest that the purified proteins may serve as bacterial receptors with broad specificity for matrix glycoproteins and that the proteins may act as carbohydrate-binding proteins.


Improved overall survival with oxaliplatin, fluorouracil, and leucovorin as adjuvant treatment in stage II or III colon cancer in the MOSAIC trial.
PURPOSE Three-year disease-free survival (DFS) was significantly improved in patients who had undergone resection with curative intent for stage II or III colon cancer who received bolus plus continuous-infusion fluorouracil plus leucovorin (LV5FU2) with the addition of oxaliplatin (FOLFOX4). Final results of the study, including 6-year overall survival (OS) and 5-year updated DFS, are reported. PATIENTS AND METHODS A total of 2,246 patients were randomly assigned to receive LV5FU2 or FOLFOX4 for 6 months. The primary end point was DFS. Secondary end points were OS and safety. Results Five-year DFS rates were 73.3% and 67.4% in the FOLFOX4 and LV5FU2 groups, respectively (hazard ratio [HR] = 0.80; 95% CI, 0.68 to 0.93; P = .003). Six-year OS rates were 78.5% and 76.0% in the FOLFOX4 and LV5FU2 groups, respectively (HR = 0.84; 95% CI, 0.71 to 1.00; P = .046); corresponding 6-year OS rates for patients with stage III disease were 72.9% and 68.7%, respectively (HR = 0.80; 95% CI, 0.65 to 0.97; P = .023). No difference in OS was seen in the stage II population. The incidence of second noncolorectal cancers was 5.5% and 6.1% in the FOLFOX4 and LV5FU2 groups, respectively. Among patients receiving oxaliplatin, the frequency of grade 3 peripheral sensory neuropathy was 1.3% 12 months after treatment and 0.7% at 48 months. CONCLUSION Adding oxaliplatin to LV5FU2 significantly improved 5-year DFS and 6-year OS in the adjuvant treatment of stage II or III colon cancer and should be considered after surgery for patients with stage III disease.


Overweight in school-aged children and its relationship with demographic and lifestyle factors: results from the WHO-Collaborative Health Behaviour in School-aged Children (HBSC) study.
To examine overweight prevalence and its association with demographic and lifestyle factors in 11-15 year olds in the HBSC 2005-2006 survey. Self-reports of height, weight, eating patterns, physical activity and sedentary behaviours were obtained from nationally representative samples in 41 countries (n=204,534). Overweight prevalence was highest in USA (28.8 %) and lowest in Latvia (7.6 %). In most countries, overweight was more prevalent in boys than girls. Overweight was consistently negatively associated with breakfast consumption and moderate to vigorous physical activity; OR range: 0.48-0.79 and 0.50-0.78, respectively. Overweight prevalence in youth remained high across the countries examined. The primary factors linked to overweight were breakfast consumption and physical activity. These data should contribute to formulating preventive programs and policies.


Anxiety sensitivity as a transdiagnostic risk factor for trajectories of adverse posttraumatic neuropsychiatric sequelae in the AURORA study.
Anxiety sensitivity, or fear of anxious arousal, is cross-sectionally associated with a wide array of adverse posttraumatic neuropsychiatric sequelae, including symptoms of posttraumatic stress disorder, depression, anxiety, sleep disturbance, pain, and somatization. The current study utilizes a large-scale, multi-site, prospective study of trauma survivors presenting to emergency departments. Hypotheses tested whether elevated anxiety sensitivity in the immediate posttrauma period is associated with more severe and persistent trajectories of common adverse posttraumatic neuropsychiatric sequelae in the eight weeks posttrauma. Participants from the AURORA study (n = 2,269 recruited from 23 emergency departments) completed self-report assessments over eight weeks posttrauma. Associations between heightened anxiety sensitivity and more severe and/or persistent trajectories of trauma-related symptoms identified by growth mixture modeling were analyzed. Anxiety sensitivity assessed two weeks posttrauma was associated with severe and/or persistent posttraumatic stress, depression, anxiety, sleep disturbance, pain, and somatic symptoms in the eight weeks posttrauma. Effect sizes were in the small to medium range in multivariate models accounting for various demographic, trauma-related, pre-trauma mental health-related, and personality-related factors. Anxiety sensitivity may be a useful transdiagnostic risk factor in the immediate posttraumatic period identifying individuals at risk for the development of adverse posttraumatic neuropsychiatric sequelae. Further, considering anxiety sensitivity is malleable via brief intervention, it could be a useful secondary prevention target. Future research should continue to evaluate associations between anxiety sensitivity and trauma-related pathology.


3D reconstruction of atomic structures from high angle annular dark field (HAADF) STEM images and its application on zeolite silicalite-1.
High-resolution transmission electron microscopy (HRTEM) has shown to be very powerful for solving three-dimensional (3D) structures of unknown crystals. HRTEM has a unique advantage over diffraction for solving structures. Crystallographic structure factor phases, which are lost in diffraction can be directly obtained from HRTEM images. For the determination of a 3D crystalline structure by HRTEM, the crystallographic structure factor amplitudes and phases extracted from HRTEM images along different zone axes are combined to reconstruct a 3D electrostatic potential map. In recent years, scanning transmission electron microscopy (STEM) has reached the atomic resolution, which is comparable to that of HRTEM. Here we show, for the first time, that the structure factor phases can be also obtained from high angle annular dark-field (HAADF)-STEM images and used for 3D reconstruction of atomic structures. This is applied to the complex zeolite structure, silicalite-1 (Formula SiO2, framework code MFI, Pnma, a = 20.090 Å, b = 19.738 Å and c = 13.142 Å). We have compared the amplitudes and phases obtained from HAADF-STEM images with those from HRTEM images.


Comparative analysis of human chromosome 7q21 and mouse proximal chromosome 6 reveals a placental-specific imprinted gene, TFPI2/Tfpi2, which requires EHMT2 and EED for allelic-silencing.
Genomic imprinting is a developmentally important mechanism that involves both differential DNA methylation and allelic histone modifications. Through detailed comparative characterization, a large imprinted domain mapping to chromosome 7q21 in humans and proximal chromosome 6 in mice was redefined. This domain is organized around a maternally methylated CpG island comprising the promoters of the adjacent PEG10 and SGCE imprinted genes. Examination of Dnmt3l(-/+) conceptuses shows that imprinted expression for all genes of the cluster depends upon the germline methylation at this putative "imprinting control region" (ICR). Similarly as for other ICRs, we find its DNA-methylated allele to be associated with trimethylation of lysine 9 on histone H3 (H3K9me3) and trimethylation of lysine 20 on histone H4 (H4K20me3), whereas the transcriptionally active paternal allele is enriched in H3K4me2 and H3K9 acetylation. Our study reveals a novel placenta-specific transcript, TFPI2, which is expressed from the maternal allele in both humans and mice. Deficiency for the histone methyltransferase EHMT2 (also known as G9A) or for the Polycomb group protein EED, involved in repressive H3K9me2 and H3K27me3 respectively, leads to biallelic expression of Tfpi2 in the extra-embryonic lineages, whereas the other genes in the cluster maintain correct imprinting. Apart from the putative ICR, however, no other promoter regions within the domain exhibited allele-specific repressive histone modifications. This unexpected general lack of repressive histone modifications suggests that this domain may utilize a different silencing mechanism as compared to other imprinted domains.


Treating acute low back pain with continuous low-level heat wrap therapy and/or exercise: a randomized controlled trial.
Restorative exercise and palliative modalities are frequently used together for the treatment of acute low back pain. However, little is known about the effects of combining these treatments. To evaluate the efficacy of combining continuous low-level heat wrap therapy with directional preference-based exercise on the functional ability of patients with acute low back pain. A randomized controlled trial was conducted at three outpatient medical facilities. One hundred individuals (age 31.2+/-10.6 years) with low back pain of less than 3 months duration. The primary outcome measure was functional ability assessed by the Multidimensional Task Ability Profile questionnaire. Secondary outcomes were disability assessed by the Roland-Morris Disability Questionnaire and pain relief assessed by a 6-point verbal rating scale. Participants were randomized to one of four groups: Heat wrap therapy alone (heat wrap, n=25); directional preference-based exercise alone (exercise, n=25); combination of heat wrap therapy and exercise (heat+exercise, n=24); or control (booklet, n=26). Treatment was administered for five consecutive days and included four visits to the study center over 1 week. At 2 days after the conclusion of treatment (Day 7), functional improvement for heat+exercise was 84%, 95%, and 175% greater than heat wrap, exercise, and booklet, respectively (p<.05). Seventy-two percent of the subjects in the heat+exercise group demonstrated a return to pre-injury function compared with 20%, 20%, and 19% for heat wrap, exercise, and booklet, respectively (p<.05). Disability reduction for heat+exercise was 93%, 139%, and 400% greater than heat wrap, exercise, and booklet, respectively (p<.05). Pain relief for heat+exercise was 70% and 143% greater than exercise and booklet, respectively (p<.05). Combining continuous low-level heat wrap therapy with directional preference-based exercise during the treatment of acute low back pain significantly improves functional outcomes compared with either intervention alone or control. Either intervention alone tends to be more effective than control.


When pits fill up: Supply and demand for safe pit-emptying services in Kisumu, Kenya.
Improving sanitation conditions in low-income communities is a major challenge for rapidly growing cities of the developing world. The expenses and logistical difficulties of extending sewerage infrastructure have focused increasing attention on the requirements for safe and cost-effective fecal sludge management services. These services, which are primarily provided by the private sector, include the collection and treatment of fecal waste from latrine pits and septic tanks. To determine the degree to which market forces can promote safe fecal sludge removal in low-income neighborhoods of Kisumu, Kenya, we compared household willingness-to-pay for formal pit emptying with the prices charged by service providers. Through surveys of 942 households and a real-money voucher trial with 646 households, we found that stated and revealed demand for formal emptying services were both low, with less than 20% of households willing to pay full market prices. Our results suggest that improving fecal sludge management in these neighborhoods via the private sector will require large subsides, ranging from 55.1-81.4 million KES (551,000-814,000 USD) annually, to address the gap between willingness-to-pay and market prices. Raising and administering subsidies of this scale will require the development of a city-wide sanitation master plan that includes investment, management, and regulatory procedures for fecal sludge management. In the absence of government investment and coordination, it is unlikely that the private sector will address safe sanitation needs in low-income areas of Kisumu.


Approximately global optimization for robust alignment of generalized shapes.
In this paper, we introduce a novel method to solve shape alignment problems. We use gray-scale "images" to represent source shapes, and propose a novel two-component Gaussian Mixture (GM) distance map representation for target shapes. This asymmetric representation is a flexible image-based representation which is able to represent different kinds of shape data, including continuous contours, unstructured sparse point sets, edge maps, and even gray-scale gradient maps. Using this representation, a new energy function based on a novel two-component Gaussian Mixture distance model is proposed. The new energy function was empirically evaluated to be a more robust shape dissimilarity metric that can be computed efficiently. Such high efficiency is essential for global optimization methods. We adopt and modify one of them, the Particle Swarm Optimization (PSO), to effectively estimate the global optimum of the new energy function. Differently from the original PSO, several new strategies were employed to make the optimization more robust and prevent it from converging prematurely. The overall performance of the proposed framework as well as the properties of each algorithmic component were evaluated and compared with those of some state-of-the-art methods. Extensive experiments and comparison performed on generalized 2D and 3D shape data demonstrate the robustness and effectiveness of the method.


Spinal arteriovenous malformations: neurological aspects and results of embolization.
Twenty consecutive patients with spinal arteriovenous malformations (AVMs) were found to have either a progressive, remitting or remitting-progressive course of disease. A characteristic progressive neurological decline was observed in 11 patients with dural fistulas and a similar decline was seen in 5 of the 9 patients with intradural AVMs. In addition to paraparesis and mixed sensory disorder, which ascended to a level of about T10, areas with spared sensation were occasionally found below this level. All cases with this progressive longitudinal myelopathy showed radiological evidence of a dilated medullary venous plexus. All patients were treated by embolization with only a low frequency of complications. Follow-up data verified a significant improvement of motor and sensory function, mainly during the first year after embolization. Cases with progressive neurological decline (dural and intradural AVMs) stopped deteriorating or improved and cases with a history of acute incidents (intradural AVMs) probably run less risk of acute deterioration.


c-Maf restrains T-bet-driven programming of CCR6-negative group 3 innate lymphoid cells.
RORγt+ group 3 innate lymphoid cells (ILC3s) maintain intestinal homeostasis through secretion of type 3 cytokines such as interleukin (IL)-17 and IL-22. However, CCR6- ILC3s additionally co-express T-bet allowing for the acquisition of type 1 effector functions. While T-bet controls the type 1 programming of ILC3s, the molecular mechanisms governing T-bet are undefined. Here, we identify c-Maf as a crucial negative regulator of murine T-bet+ CCR6- ILC3s. Phenotypic and transcriptomic profiling of c-Maf-deficient CCR6- ILC3s revealed a hyper type 1 differentiation status, characterized by overexpression of ILC1/NK cell-related genes and downregulation of type 3 signature genes. On the molecular level, c-Maf directly restrained T-bet expression. Conversely, c-Maf expression was dependent on T-bet and regulated by IL-1β, IL-18 and Notch signals. Thus, we define c-Maf as a crucial cell-intrinsic brake in the type 1 effector acquisition which forms a negative feedback loop with T-bet to preserve the identity of CCR6- ILC3s.


Early antiretroviral therapy during primary HIV-1 infection results in a transient reduction of the viral setpoint upon treatment interruption.
Long-term benefits of combination antiretroviral therapy (cART) initiation during primary HIV-1 infection are debated. The evolution of plasma HIV-RNA (432 measurements) and cell-associated HIV-DNA (325 measurements) after cessation of cART (median exposure 18 months) was described for 33 participants from the Zurich Primary HIV Infection Study using linear regression and compared with 545 measurements from 79 untreated controls with clinically diagnosed primary HIV infection, respectively a known date for seroconversion. On average, early treated individuals were followed for 37 months (median) after cART cessation; controls had 34 months of pre-cART follow-up. HIV-RNA levels one year after cART interruption were -0.8 log₁₀ copies/mL [95% confidence interval -1.2;-0.4] lower in early treated patients compared with controls, but this difference was no longer statistically significant by year three of follow-up (-0.3 [-0.9; 0.3]). Mean HIV-DNA levels rebounded from 2 log₁₀ copies [1.8; 2.3] on cART to a stable plateau of 2.7 log₁₀ copies [2.5; 3.0] attained 1 year after therapy stop, which was not significantly different from cross-sectional measurements of 9 untreated members of the control group (2.8 log₁₀ copies [2.5; 3.1]). The rebound dynamics of viral markers after therapy cessation suggest that early cART may indeed limit reservoir size of latently infected cells, but that much of the initial benefits are only transient. Owing to the non-randomized study design the observed treatment effects must be interpreted with caution.


Preparation and Characterization of Griseofulvin Solid Dispersions.
Amorphous solid dispersion (SD) technique has been used for improving the solubility and bioavailability of poorly water-soluble compounds. However, the stability of these SD is a concern due to the high propensity of metastable amorphous form to convert into its stable crystalline form. In this study, the stability of SD of griseofulvin (GSV), a high crystallization tendency compound, was evaluated in the presence of commonly used Food and Drug Administration-approved hydrophilic polymers such as polyvinylpyrrolidone (PVP), hydroxypropyl methylcellulose, Eudragits, and polyethylene glycol. Physical mixtures (PM) and SD were prepared at various drug to polymer ratios (3:1, 1:1, 1:2, 1:9 w/w) using the solvent evaporation technique. PM and SD were characterized by powder X-ray diffractometer and modulated differential scanning calorimetry to confirm the formation of amorphous dispersions and to understand the crystallization inhibition effectiveness of polymers. Infrared (IR), Raman spectroscopy, and molecular modeling were used to understand the role of interaction between GSV and polymers for stabilization of dispersions. We observed that most polymers are ineffective in stabilizing the amorphous form of GSV at low concentrations, that is, 3:1, 1:1, and 1:2 w/w except for PVP K-90, which was effective at a concentration of 1:1 w/w. Slight peak shifts were observed in IR and Raman; however, these shifts were inconclusive of molecular interactions between GSV and hydrophilic polymers. GSV's high crystallization tendency coupled with a lack of hydrogen bond acceptors/donors hinders the formation of stable SD, and thus, a higher polymer concentration is a prerequisite for their long-term stability.


Molecular mechanism of lipopeptide presentation by CD1a.
CD1a is expressed on Langerhans cells (LCs) and dendritic cells (DCs), where it mediates T cell recognition of glycolipid and lipopeptide antigens that contain either one or two alkyl chains. We demonstrate here that CD1a-restricted T cells can discriminate the peptide component of didehydroxymycobactin lipopeptides. Structure analysis of CD1a cocrystallized with a synthetic mycobactin lipopeptide at 2.8 A resolution further reveals that the single alkyl chain is inserted deep within the A' pocket of the groove, whereas its two peptidic branches protrude along the F' pocket to the outer, alpha-helical surface of CD1a for recognition by the TCR. Remarkably, the cyclized lysine branch of the peptide moiety lies in the shallow F' pocket in a conformation that closely mimics that of the alkyl chain in the CD1a-sulfatide structure. Thus, this structural study illustrates how a single chain lipid can be presented by CD1 and that the peptide moiety of the lipopeptide is recognized by the TCR.


Impact of oral hygiene involving toothbrushing versus chlorhexidine in the prevention of ventilator-associated pneumonia: a randomized study.
Nosocomial pneumonia has correlated to dental plaque and to oropharynx colonization in patients receiving mechanical ventilation. The interruption of this process, by preventing colonization of pathogenic bacteria, represents a potential procedure for the prevention of ventilator-associated pneumonia (VAP). The study design was a prospective, randomized trial to verify if oral hygiene through toothbrushing plus chlorhexidine in gel at 0.12% reduces the incidence of ventilatior-associated pneumonia, the duration of mechanical ventilation, the length of hospital stay and the mortality rate in ICUs, when compared to oral hygiene only with chlorhexidine, solution of 0.12%, without toothbrushing, in adult individuals under mechanical ventilation, hospitalized in Clinical/Surgical and Cardiology Intensive Care Units (ICU). The study protocol was approved by the Ethical Committee of Research of the Health Sciences Center of the Federal University of Pernambuco - Certificate of Ethical Committee Approval (CAAE) 04300012500005208. Because it was a randomized trial, the research used CONSORT 2010 checklist criteria. Seven hundred sixteen patients were admitted into the ICU; 219 fulfilled the criteria for inclusion and 213 patients were included; 108 were randomized to control group and 105 to intervention group. Toothbrushing plus 0.12% chlorhexidine gel demonstrated a lower incidence of VAP throughout the follow up period, although the difference was not statistically significant (p = 0.084). There was a significant reduction of the mean time of mechanical ventilation in the toothbrushing group (p = 0.018). Regarding the length of hospital stay in the ICU and mortality rates, the difference was not statistically significant (p = 0.064). The results obtained showed that, among patients undergoing toothbrushing there was a significant reduction in duration of mechanical ventilation, and a tendency to reduce the incidence of VAP and length of ICU stay, although without statistical significance. Retrospectively registered in the Brazilian Clinical Trials Registry (Registro Brasileiro de Ensaios Clínicos) - RBR-4TWH4M (4 September 2016).


Local inhibition and long-range enhancement of Dpp signal transduction by Sog.
Extracellular gradients of signalling molecules can specify different thresholds of gene activity in development. A gradient of Decapentaplegic (Dpp) activity subdivides the dorsal ectoderm of the Drosophila embryo into amnioserosa and dorsal epidermis. The proteins Short gastrulation (Sog) and Tolloid (Tld) are required to shape this gradient. Sog has been proposed to form an inhibitory complex with either Dpp or the related ligand Screw, and is subsequently processed by the protease Tld. Paradoxically, Sog appears to be required for amnioserosa formation, which is specified by peak Dpp signalling activity. Here we show that the misexpression of sog using the even-skipped stripe-2 enhancer redistributes Dpp signalling in a mutant background in which dpp is expressed throughout the embryo. Dpp activity is diminished near the Sog stripe and peak Dpp signalling is detected far from this stripe. However, a tethered form of Sog suppresses local Dpp activity without augmenting Dpp activity at a distance, indicating that diffusion of Sog may be required for enhanced Dpp activity and consequent amnioserosa formation. The long-distance stimulation of Dpp activity by Sog requires Tld, whereas Sog-mediated inhibition of Dpp does not. The heterologous Dpp inhibitor Noggin inhibits Dpp signalling but fails to augment Dpp activity. These results suggest an unusual strategy for generating a gradient threshold of growth-factor activity, whereby Sog and its protease specify peak Dpp signalling far from a localized source of Sog.


Complete sequence analysis of mitochondrial DNA and telomere length in aplastic anemia.
The present study was primarily undertaken to examine the hypothesis that mitochondrial DNA (mtDNA) mutations and telomere length may be associated with aplastic anemia (AA). Our study included a single institution analysis of 40 patients presenting with AA first diagnosed at the Affiliated Hospital of Shandong, University of Traditional Chinese Medicine between 2010 and 2013. Bone marrow and oral epithelial samples were collected from patients with AA (n=40) for mtDNA mutation and telomere length determinations. Bone marrow specimens were collected from 40 healthy volunteers as controls for the examination of telomere length. The mitochondrial genome was amplified by polymerase chain reaction (PCR), and the products were used for sequencing and analysis. We detected 146 heteroplasmic mutations in 18 genes from 40 patients with AA, including 39 silent mutations and 28 frameshift mutations. We used the gamma globin gene (HBG) as the control gene in real-time PCR to survey the relative telomere length measurements of the patients with AA and the healthy volunteers. Telomere length was expressed as the relative T/S value. We observed a negative correlation between the mtDNA non-silent mutation and the white blood cell (WBC) count, hemoglobin and platelet count. Of note, there was a positive correlation between the relative T/S value and WBC count, hemoglobin and platelet count, and a negative correlation between the non-silent mutation and the relative T/S value. We conclude that the functional impairment of the mitochondrial respiratory chain induced by mutation and telomere length shortening may play an important role in the process of hematopoietic failure in patients with AA. Additionally, mtDNA mutations and telomere length shortening influenced each other.


1.5 μm polarization entanglement generation based on birefringence in silicon wire waveguides.
In this Letter, the 1.5 μm polarization entanglement generation is realized in a silicon wire waveguide utilizing its birefringence. In this scheme, two orthogonal polarized correlated states are generated by scalar processes of spontaneous four-wave mixing (SFWM) in the quasi-transverse electrical and quasi-transverse magnetic modes, respectively. Meanwhile, the vector processes of SFWM are suppressed by the group birefringence in the waveguide. The maximum polarization entangled state is generated by optimizing the pump polarization, which is demonstrated by the experiments of two-photon interference and polarization indistinguishability at one side. The fringe visibilities of two-photon interferences are 96.8±4.7% and 86.0±3.7% under two nonorthogonal polarization detection settings, respectively. This scheme provides a simple way to realize silicon integrated sources for 1.5 μm polarization entanglement generation.


Transition Metal-Catalyzed Intermolecular Cascade C-H Activation/Annulation Processes for the Synthesis of Polycycles.
Polycycles are abundantly present in numerous advanced chemicals, functional materials, bioactive molecules and natural products. However, the strategies for the synthesis of polycycles are limited to classical reactions and transition metal-catalyzed cross-coupling reactions, requiring pre-functionalized starting materials and lengthy synthetic operations. The emergence of novel approaches shows great promise for the fields of organic/medicinal/materials chemistry. Among them, transition metal-catalyzed C-H activation followed by intermolecular annulation reactions prevail, due to their straightforward manner with high atom- and step-economy, providing rapid, concise and efficient methods for the construction of diverse polycycles. Several strategies have been developed for the synthesis of polycycles, relying on sequential multiple C-H activation/annulation, or combination of C-H activation/annulation and further interaction with a proximal group, or merger of C-H activation with a cycloaddition reaction, or in situ formation of the directing group. These are attractive, efficient, step- and atom-economic methods starting from commercially available materials. This Minireview will provide an introduction to transition metal-catalyzed C-H activation for the synthesis of polycycles, helping researchers to discover indirect connections and reveal hidden opportunities. It will also promote the discovery of novel synthetic strategies relying on C-H activation.


Assessment of functional capacity before major non-cardiac surgery: an international, prospective cohort study.
Functional capacity is an important component of risk assessment for major surgery. Doctors' clinical subjective assessment of patients' functional capacity has uncertain accuracy. We did a study to compare preoperative subjective assessment with alternative markers of fitness (cardiopulmonary exercise testing [CPET], scores on the Duke Activity Status Index [DASI] questionnaire, and serum N-terminal pro-B-type natriuretic peptide [NT pro-BNP] concentrations) for predicting death or complications after major elective non-cardiac surgery. We did a multicentre, international, prospective cohort study at 25 hospitals: five in Canada, seven in the UK, ten in Australia, and three in New Zealand. We recruited adults aged at least 40 years who were scheduled for major non-cardiac surgery and deemed to have one or more risk factors for cardiac complications (eg, a history of heart failure, stroke, or diabetes) or coronary artery disease. Functional capacity was subjectively assessed in units of metabolic equivalents of tasks by the responsible anaesthesiologists in the preoperative assessment clinic, graded as poor (<4), moderate (4-10), or good (>10). All participants also completed the DASI questionnaire, underwent CPET to measure peak oxygen consumption, and had blood tests for measurement of NT pro-BNP concentrations. After surgery, patients had daily electrocardiograms and blood tests to measure troponin and creatinine concentrations until the third postoperative day or hospital discharge. The primary outcome was death or myocardial infarction within 30 days after surgery, assessed in all participants who underwent both CPET and surgery. Prognostic accuracy was assessed using logistic regression, receiver-operating-characteristic curves, and net risk reclassification. Between March 1, 2013, and March 25, 2016, we included 1401 patients in the study. 28 (2%) of 1401 patients died or had a myocardial infarction within 30 days of surgery. Subjective assessment had 19·2% sensitivity (95% CI 14·2-25) and 94·7% specificity (93·2-95·9) for identifying the inability to attain four metabolic equivalents during CPET. Only DASI scores were associated with predicting the primary outcome (adjusted odds ratio 0·96, 95% CI 0·83-0·99; p=0·03). Subjectively assessed functional capacity should not be used for preoperative risk evaluation. Clinicians could instead consider a measure such as DASI for cardiac risk assessment. Canadian Institutes of Health Research, Heart and Stroke Foundation of Canada, Ontario Ministry of Health and Long-Term Care, Ontario Ministry of Research, Innovation and Science, UK National Institute of Academic Anaesthesia, UK Clinical Research Collaboration, Australian and New Zealand College of Anaesthetists, and Monash University.


Evolution of the Telomere-Associated Protein POT1a in Arabidopsis thaliana Is Characterized by Positive Selection to Reinforce Protein-Protein Interaction.
Gene duplication is a major driving force in genome evolution. Here, we explore the nature and origin of the POT1 gene duplication in Arabidopsis thaliana. Protection of Telomeres (POT1) is a conserved multifunctional protein that modulates telomerase activity and its engagement with telomeres. Arabidopsis thaliana encodes two divergent POT1 paralogs termed AtPOT1a and AtPOT1b. AtPOT1a positively regulates telomerase activity, whereas AtPOT1b is proposed to negatively regulate telomerase and promote chromosome end protection. Phylogenetic analysis uncovered two independent POT1 duplication events in the plant kingdom, including one at the base of Brassicaceae. Tests for positive selection implemented in PAML revealed that the Brassicaceae POT1a lineage experienced positive selection postduplication and identified three amino acid residues with signatures of positive selection. A sensitive and quantitative genetic complementation assay was developed to assess POT1a function in A. thaliana. The assay showed that AtPOT1a is functionally distinct from single-copy POT1 genes in other plants. Moreover, for two of the sites with a strong signature of positive selection, substitutions that swap the amino acids in AtPOT1a for residues found in AtPOT1b dramatically compromised AtPOT1a function in vivo. In vitro-binding studies demonstrated that all three sites under positive selection specifically enhance the AtPOT1a interaction with CTC1, a core component of the highly conserved CST (CTC1/STN1/TEN1) telomere protein complex. Our results reveal a molecular mechanism for the role of these positively selected sites in AtPOT1a. The data also provide an important empirical example to refine theories of duplicate gene retention, as the outcome of positive selection here appears to be reinforcement of an ancestral function, rather than neofunctionalization. We propose that this outcome may not be unusual when the duplicated protein is a component of a multisubunit complex whose function is in part specified by other members.


Randomised controlled trial of screening for Chlamydia trachomatis to prevent pelvic inflammatory disease: the POPI (prevention of pelvic infection) trial.
To determine whether screening and treating women for chlamydial infection reduces the incidence of pelvic inflammatory disease over the subsequent 12 months. Randomised controlled trial. Common rooms, lecture theatres, and student bars at universities and further education colleges in London. 2529 sexually active female students, mean age 21 years (range 16-27). Participants completed a questionnaire and provided self taken vaginal swabs, with follow-up after one year. Samples were randomly allocated to immediate testing and treatment for chlamydial infection, or storage and analysis after a year (deferred screening controls). Incidence of clinical pelvic inflammatory disease over 12 months. Baseline prevalence of chlamydia was 5.4% (68/1254) in screened women and 5.9% (75/1265) in controls. 94% (2377/2529) of women were followed up after 12 months. The incidence of pelvic inflammatory disease was 1.3% (15/1191) in screened women compared with 1.9% (23/1186) in controls (relative risk 0.65, 95% confidence interval 0.34 to 1.22). Seven of 74 control women (9.5%, 95% confidence interval 4.7% to 18.3%) who tested positive for chlamydial infection at baseline developed pelvic inflammatory disease over 12 months compared with one of 63 (1.6%) screened women (relative risk 0.17, 0.03 to 1.01). However, most episodes of pelvic inflammatory disease occurred in women who tested negative for chlamydia at baseline (79%, 30/38). 22% (527/2377) of women reported being tested independently for chlamydia during the trial. Although some evidence suggests that screening for chlamydia reduces rates of pelvic inflammatory disease, especially in women with chlamydial infection at baseline, the effectiveness of a single chlamydia test in preventing pelvic inflammatory disease over 12 months may have been overestimated. Trial registration ClinicalTrials.gov NCT00115388.


Use of monoclonal antibodies prepared against Schistosoma mansoni hatching fluid antigens for demonstration of Schistosoma haematobium circulating egg antigens in urine.
A panel of 17 monoclonal antibodies (MAbs) against Schistosoma soluble egg antigens (SEAs) was produced from BALB/c mice immunized with antigens secreted/excreted by Schistosoma mansoni eggs. In this study, we demonstrate that 16 MAbs were reactive with S. haematobium SEA in addition to S. mansoni SEA. The MAbs were tested as potential immunodiagnostic reagents in a homologous sandwich ELISA format to detect circulating soluble egg antigens (CSEAs) in serum and urine samples of S. mansoni- or S. haematobium-infected individuals. When samples of S. mansoni-infected individuals were tested, none of these MAbs performed as good as the previously described S. mansoni-specific 114-5B1-A and 114-4D12-A MAbs. However, 11 MAbs (of the IgM isotype) detected CSEA in urine samples of S. haematobium-infected individuals. Three MAbs, 290-2E6-A, 291-3D5-A, and 291-5D5-A, were selected for a pilot study with 47 urine samples of S. haematobium-infected individuals from Kenya. The CSEA levels detected with each of these ELISAs showed a significant correlation with urinary egg counts (Spearman rho > 0.37, P < 0.01) and with each other (Spearman rho > 0.74, P < 0.001). Based on the 92% specificity and 90% sensitivity of the assay, the ELISA using MAb 290-2E6-A was found to be the most promising assay for immunodiagnosis of S. haematobium infections.


Hepatic disorders in patients treated with COX-2 selective inhibitors or nonselective NSAIDs: a case/noncase analysis of spontaneous reports.
Hepatic adverse events associated with the use of nonaspirin drugs and NSAIDs are uncommon, but the widespread use of these drugs may impact public health. We conducted a case/noncase analysis of spontaneous reports to compare the hepatic safety profile of cyclooxygenase (COX)-2 selective inhibitors with that of nonselective NSAIDs. This case/noncase analysis was conducted using the US Food and Drug Administration Freedom of Information (FDA/FOI) database (through quarter 1, 2003) and the World Health Organization Uppsala Monitoring Centre (WHO/UMC) database (through quarter 3, 2003). Council for International Organizations of Medical Sciences and WHO Adverse Reaction Terminology preferred terms were used to classify hepatic disorders with broad and specific case definitions. After reports involving established hepatotoxic drugs (bromfenac, nimesulide, sulindac) were excluded, the proportion of reports (PRs) of each case definition was calculated for each NSAID. Crude and adjusted reporting odds ratios (RORs) were used to compare the overall proportions of hepatic disorders and hepatic failure of celecoxib and rofecoxib versus nonselective NSAIDs. A total of 158,539 and 185,253 reports of NSAIDs were identified in the FDA/FOI and WHO/UMC databases and 25% and 16%, respectively, involved other hepatotoxic drugs. The PRs of hepatic disorders for all COX-2 selective inhibitors and non-selective NSAIDs were 3.0% in the FDA/FOI database and 2.7% in the WHO/UMC database. In the FDA/FOI and WHO/UMC databases, respectively, mmesulide (16.7% and 14.4%), bromfenac (12.0% and 20.7%), diclofenac (8.1% and 4.7%), and sulindac (6.1 % and 9.9%) were reported to be associated with higher proportions of overall hepatic disorders compared with those of other NSAIDs. Crude and adjusted RORs for the prevalences of overall hepatic disorders and hepatic failure with celecoxib and rofecoxib versus the other NSAIDs were <1 (indicating that the proportion was not higher than that of the comparator) in both databases. The interpretation of the results was unchanged when bromfenac, nimesulide, and sulindac were excluded from the analysis. In this case/noncase analysis, bromfenac, nimesulide, sulindac, and diclofenac had higher proportions of reports of hepatic disorders compared with those of other NSAIDs in the FDA/FOI and WHO/UMC databases. The analysis did not raise a safety concern for celecoxib or rofecoxib versus NSAIDs for overall hepatic disorders and hepatic failure.


Phenomenology of COPD: interpreting phenotypes with the ECLIPSE study.
The Evaluation of COPD Longitudinally to Identify Predictive Surrogate End-points (ECLIPSE) study was a large 3-year observational multicentre international study aimed at defining COPD phenotypes and identifying biomarkers and/or genetic parameters that help to predict disease progression. The study has contributed to a better understanding of COPD heterogeneity, with the characterization of clinically important subtypes/phenotypes of patients, such as the frequent exacerbators or patient with persistent systemic inflammation, who may have different prognosis or treatment requirements. Because of the big amount of information that is starting to be produced from metabolomic, proteomic and genomic approaches, one of the biggest challenges is the integration of data in a biological prospective such as clinical prognosis and response to medicinal products. In this article we highlight some of the progress in phenotyping the heterogeneity of the disease that have been made thanks to the analyses of this longitudinal study.


A systematic review and meta-analysis of adverse outcomes following non-buried versus buried Kirschner wires for paediatric lateral condyle elbow fractures.
Fractures of the lateral condyle of the humerus in children are a common injury. If displaced or unstable they may require surgical reduction and fixation with Kirschner wires (K-wires). K-wires are placed using either an open or closed technique. The decision to bury or leave the ends extending through the skin is surgeon-dependent and based on factors including post-operative infection risk, bony union and ease of wire removal. We performed a systematic review and meta-analysis of non-buried versus buried K-wires for lateral condyle elbow fractures in children in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses methodology. A comprehensive search strategy included Medline, Embase and CINAHL via NICE Evidence from database inception to June 2017. Two authors independently reviewed, included or excluded articles, extracted data and assessed for quality with the ROBINS-I tool. We performed direct comparison meta-analysis for all adverse events, post-operative infection and failure of bony union. Three studies were analysed comprising of 434 participants. There was a significantly reduced relative risk of adverse events in the non-buried group, equating to approximately 45% reduced risk (RR 0.55, 95% confidence interval 0.34 to 0.88). There were no significant differences in risk of post-operative infection or failure of bony union. All three cost-analyses in the included studies observed savings with non-buried K-wires. Non-buried K-wires for lateral condyle elbow fractures convey a lower risk of adverse events and may be more cost-effective compared with buried K-wires. Non-buried K-wires do not appear to increase the risk of infection or failure of bony union. These findings are limited by a high risk of bias due to inherent methodological flaws in the design of included studies.


Plk1 phosphorylation of orc2 and hbo1 contributes to gemcitabine resistance in pancreatic cancer.
Although gemcitabine is the standard chemotherapeutic drug for treatment of pancreatic cancer, almost all patients eventually develop resistance to this agent. Previous studies identified Polo-like kinase 1 (Plk1) as the mediator of gemcitabine resistance, but the molecular mechanism remains unknown. In this study, we show that Plk1 phosphorylation of Orc2 and Hbo1 mediates the resistance to gemcitabine. We show that the level of Plk1 expression positively correlates with gemcitabine resistance, both in pancreatic cancer cells and xenograft tumors. Overexpression of Plk1 increases gemcitabine resistance, while inhibition of Plk1 sensitizes pancreatic cancer cells to gemcitabine treatment. To validate our findings, we show that inhibition of Plk1 sensitizes tumors to gemcitabine treatment in a mouse xenograft study. Mechanistically, we find that Plk1 phosphorylation of Orc2 maintains DNA replication on gemcitabine treatment. Furthermore, Plk1 phosphorylation of Hbo1 transcriptionally increases cFos expression and consequently elevates its target multidrug resistance 1 (MDR1), which was previously reported to confer chemotherapeutic drug resistance. Knockdown of cFos or MDR1 sensitizes gemcitabine-resistant cells to gemcitabine treatment. Finally, pancreatic cancer cells expressing Plk1-unphosphorylatable mutants of Orc2 or Hbo1 are more sensitive to gemcitabine than cells expressing wild-type Orc2 or Hbo1. In short, our study provides a mechanism for Plk1-mediated gemcitabine resistance, suggesting that Plk1 is a promising target for treatment of gemcitabine-resistant pancreatic cancer.


Structural variety beyond appearance: high-pressure phases of CrB4 in comparison with FeB4.
Employing particle swarm optimization (PSO) combined with first-principles calculations, we systemically studied high-pressure behaviors of hard CrB4. Our predictions reveal a distinct structural evolution under pressure for CrB4 despite having the same initial structure as FeB4. CrB4 is found to adopt a new P2/m structure above 196 GPa, another Pm structure at a pressure range of 261-294 GPa and then a Pmma structure beyond 294 GPa. Instead of puckering boron sheets in the initial structure, the high-pressure phases have planar boron sheets with different motifs upon compression. Comparatively, FeB4 prefers an I41/acd structure over 48 GPa with tetrahedron B4 units and a P213 structure above 231 GPa having equilateral triangle B3 units. Significantly, CrB4 exhibits persistent metallic behavior in contrast with the semiconducting features of FeB4 upon compression. The varied pressure response of hard tetraborides studied here is of importance for understanding boron-rich compounds and designing new materials with superlative properties.


Cross-reactive cytotoxic responses. H-2 restricted are more specific than anti-H-2 responses.
Cross-reactive T-cell cytotoxicity is seen when cytotoxic responses are generated in mixed lymphocyte cultures either between mouse strans which differ at the major histocompatibility complex, H-2, or between H-2b mutant strains and the strain from which they were derived. This cross-reactivity can be measured with [51Cr] labeled target cells from a number of different H-2 haplotypes, and the pattern of cross-reaction indicates that the target antigens are unlikely to be any of the serologically defined public specificities. In contrast, the specificity of H-2 restricted cytotoxic responses, such as that to the male-specific antigen, H-Y, is exquisite, and male cells from strains of mice carrying H-2 haplotypes other than the responder have never been found to act as appropriate targets. The contrast between the specificity of anti-H-2 and H-2 restricted responses may argue for a greater idiotypic homogeneity of the cells makiing H-2 restricted responses, and the greater specificity of these responses may be necessary for their biological function.


In vivo anti-arthritic activity of Bauhinia purpurea Linn. Bark Extract.
Bauhinia purpurea (BP) Linn. (Caesalpiniaceae) is a plant of great medicinal importance and has been used since ancient times for treating many inflammatory conditions including arthritis. This study investigates the anti-arthritic potential of the hydroalcoholic extract from the stem bark of BP. The anti-inflammatory and anti-arthritic activity of BP at various doses was used to evaluate its anti-inflammatory activity and anti-arthritic activity. Serum of arthritic rats was collected at day 21 for detecting serum cytokines level and to evaluate the effect of BP on its serum level. Furthermore, the safety of BP was evaluated in acute (5 days) and subacute (28 days) toxicity study in rats. There was a significant inhibition (P < 0.01) in paw edema at a different time scale with different doses of BP (50, 100, and 200 mg/kg). BP also demonstrated dose-dependent anti-arthritic activity on all observation days (3, 7, 14, and 21). In addition, there was also a significant decrease (P < 0.01) in oxidative stress markers, circulating pro-inflammatory cytokine (tumor necrosis factor alpha from 45.91 to 37.44, interleukin-1 (IL-1) β from 18.24 to 16.06, and IL-6 from 69.77 to 58.44) and an increase in anti-inflammatory cytokine (IL-10 from 8.07 to 12.07) levels. BP was found to be safe with an oral LD50 value of >2 g/kg in acute toxicity study and also no toxicological effect was observed in the oral subacute toxicity study. This study demonstrates that BP bark possesses anti-arthritic activity potential and confirm its folklore use in the treatment of inflammatory conditions.


Clinical outcome of autologous bone marrow aspirates concentrate (BMAC) injection in degenerative arthritis of the knee.
As a treatment method of degenerative arthritis of knee, this study evaluated the clinical efficacy of the intra-articular injection of autologous bone marrow aspirates concentrate (BMAC) with adipose tissue. Between April 2011 and May 2012, 41 patients (75 knees) who were diagnosed as a degenerative knee arthritis and underwent the BMAC injection with adipose tissue were included in this study. Mean age was 60.7 years old (ranged 53-80). Kellgren-Lawrence grade was used for assessing radiologic degree of osteoarthritis; there were each 12, 24, 33, and 6 cases of grade I, II, III, and IV. At preoperative and postoperative 3, 6, and 12 months, pain score using visual analogue scale (VAS) and functional scales were used for evaluation. After the procedure, mean VAS score was decreased from 7.0 preoperatively to 4.1, 3.5, and 3.3 postoperatively 3, 6, and 12 months. And functional scores were also improved; International Knee Documentation Committee score (from 37.7 preoperatively to 59.3, 66.3, 69.3 postoperatively), SF-36 health score (from 31.5 to 43.5, 45.6, 47.7), knee and osteoarthritis outcome score (from 43.1 to 64.9, 68.5, 70.6), Lysholm Knee Questionnaire (from 37.3 to 65.4, 68.6, 71.0) were all increased after the procedure. When classified according to K-L grade, the improvement of VAS score in grade IV group was 8.2 preoperatively to 5.5, 5.3, and 5.7 postoperatively, which was significantly poorer than those of grade I-III groups. In the knee functional scales, similar pattern was checked. BMAC injection significantly improved both knee pain and functions in the patients with degenerative arthritis of knee. Also, the injection would be more effective in early to moderate phases.


Light-induced variability in development of forensically important blow fly Phormia regina (Diptera: Calliphoridae).
The use of the postmortem interval (PMI) in practical applications of forensic entomology is based upon developmental data of blow flies (Diptera: Calliphoridae) generated under controlled environmental conditions. Careful review of the published forensic entomology data sets showed that experimental (environmental) parameters differed between studies. Despite the differences in study design, there are no empirical data on the effect of photoperiod on blow fly development; yet, photoperiod has been shown to alter some insect development and behavior among noncalliphorids. Consequently, will differences in photoperiod alter the developmental times of calliphorids, and thereby alter PMI estimates? To answer this question, we used a replicated design with precise temperature measurement to examine the effects of photoperiod on the forensically important blow fly Phormia regina (Meigen). We concluded that inaccurate temperature recordings by using set-chamber temperatures over rearing-container temperatures would have overshadowed any affect light had on development. Second, constant light increased variation in overall adult developmental time and significantly delayed development compared with cyclic light. Finally, not accounting for delayed development induced by photoperiod underestimated the initial empirical estimate of the PMI. These sources of variation need to be included in forensic estimates because this variation can compromise predictions of PMI based upon current data sets. Without pinpointing optimal photoperiods with which to test development, we must assume that potentially large sources of variability exist within current estimates of the PMI.


[Trends in HIV prevalence and associated risk behaviors in female sex workers in Catalonia (Spain)].
To describe trends in HIV prevalence, sexual risk behaviors and other sexual health indicators in female sex workers (SW) from 2005 to 2011. Cross-sectional studies were conducted biennially among SW recruited in Catalonia (Spain) in the street, flats and clubs (n = 400). In addition to an anonymous questionnaire, oral fluid samples were collected to estimate HIV prevalence. Linear trends in proportions were assessed by the Mantel test. The percentage of SW who used condoms inconsistently with clients (past 6 months) increased from 5.1% in 2005 to 9.9% in 2011 (p = 0.005); this percentage also increased with steady partners (86.2% in 2005 to 94.4% in 2011, p = 0.002). The prevalence of self-reported sexually transmitted infections (STI) increased from 14% in 2005 to 20.6% in 2011 (p = 0.001). The prevalence of HIV held constant over the years (about 2%), being higher in Spanish women (14.7% in 2011). The prevalence of HIV in SW remained stable over the years, being higher in Spanish SW. The increase in unprotected sex and in the prevalence of self-reported STI among SW from 2005 to 2011 highlights a possible relaxation in preventive behaviors in this group. Interventions to reduce the transmission of these infections, as well as unwanted pregnancies, should continue in Catalonia in SW.


Zirconia in dentistry: part 2. Evidence-based clinical breakthrough.
An ideal all-ceramic restoration that conforms well and demonstrates enhanced biocompatibility, strength, fit, and esthetics has always been desirable in clinical dentistry. However, the inherent brittleness, low flexural strength, and fracture toughness of conventional glass and alumina ceramics have been the main obstacles for extensive use. The recent introduction of zirconia-based ceramics as a restorative dental material has generated considerable interest in the dental community, which has been expressed with extensive industrial, clinical, and research activity. Contemporary zirconia powder technology contributes to the fabrication of new biocompatible all-ceramic restorations with improved physical properties for a wide range of promising clinical applications. Especially with the development of computer-aided design (CAD)/computer-aided manufacturing (CAM) systems, high-strength zirconia frameworks can be viable for the fabrication of full and partial coverage crowns, fixed partial dentures, veneers, posts and/or cores, primary double crowns, implant abutments, and implants. Data from laboratory and clinical studies are promising regarding their performance and survival. However, clinical data are considered insufficient and the identified premature complications should guide future research. In addition, different zirconia-based dental auxiliary components (i.e., cutting burs and surgical drills, extra-coronal attachments and orthodontic brackets) can also be technologically feasible. This review aims to present and discuss zirconia manufacturing methods and their potential for successful clinical application in dentistry.


Alterations in thyroid function in female Sprague-Dawley rats following chronic treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin.
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a multisite carcinogen. Although the hepatocarcinogenic actions of TCDD have received the most attention, it has been demonstrated in several rodent carcinogenicity bioassays that TCDD causes a dose-related increase in thyroid follicular cell adenomas and carcinomas. The purpose of the present experiment was to investigate the dose-response relationship for thyroid function alterations in female Sprague-Dawley rats following chronic treatment with TCDD. TCDD was administered via oral gavage biweekly for 30 weeks at average daily equivalent doses of 0.1-125 ng/kg/day, thereby more than encompassing the dose range historically used in previous TCDD rodent bioassays. The endpoints examined include serum levels of thyroxine (T4), triiodothyronine (T3), and thyroid-stimulating hormone (TSH). In addition, the induction of the dioxin-responsive genes UDP-glucuronosyltransferase-1 (UGT1) and cytochrome P450 1A1 (CYP1A1) in liver were measured using reverse-transcriptase-polymerase chain reaction (RT-PCR). In agreement with previous hypotheses, TCDD appears to alter thyroid function via a secondary mechanism, namely increased excretion of T4-glucuronide resulting from TCDD induction of UGT1. The observed follicular cell hyperplasia and hypertrophy are consistent with the observed elevated TSH levels and may represent the early stages in the progression of thyroid carcinogenesis. Therefore, TCDD induces alterations in thyroid hormone function, probably as a result of chronic perturbations of liver-pituitary-thyroid axis.


Evaluation of correction factors for transmittance measurements in single-beam integrating spheres.
An integrating sphere for transmittance measurements at normal and oblique angles of incidence has been constructed. The sphere is a single-beam instrument that uses a small-area silicon diode as the detector. The entry port is only 0.37% of the total wall area and has an oblong shape to permit measurements at high angles of incidence for scattering samples. A small beam size has been made possible by using a low-noise preamplifier system for the detector circuit. The oblong port shape and a small beam size make it possible to perform simulated double-beam measurements at near-normal incidence. Modified correction factors for the sample reflectance have been derived. Special attention has been paid to the separation into a diffuse and a specular component of the transmitted light. Results have been compared with the results of measurements on a double-beam instrument, and the correction factors for specular and diffuse samples have been experimentally verified. The importance of using the right correctionfactors for different types of samples has been evaluated together with the influence of the sphere parameters.


Drug Target Discovery Methods In Targeting Neurotropic Parasitic Amoebae.
Neurotropic parasitic amoebal infections have imposed an enormous challenge to chemotherapy in patients who fall victims to the infections caused by them. Conventional antibiotics that are given to treat these infections have a low patient compliance because of the serious adverse effects that are associated with their use. Additionally, the growing incidence of the development of drug resistance by the neurotropic parasites like Naegleria fowleri, Balamuthia mandrillaris, and Acanthamoeba spp has made the drug therapy more challenging. Recent studies have reported some cellular targets in the neurotropic parasitic Acanthamoeba that are used as receptors by human neurotransmitters like acetylcholine. This Viewpoint attempts to highlight the novel methodologies that use drug assays and structural modeling to uncover cellular targets of diverse groups of drugs and the safety issues of the drugs proposed for their use in brain infections caused by the neurotropic parasitic amoebae.


Age-related task sensitivity of frontal EEG entropy during encoding predicts retrieval.
Age-related declines in memory may be due in part to changes in the complexity of neural activity in the aging brain. Electrophysiological entropy provides an accessible measure of the complexity of ongoing neural activity. In the current study, we calculated the permutation entropy of the electroencephalogram (EEG) during encoding of relevant (to be learned) and irrelevant (to be ignored) stimuli by younger adults, older adults, and older cognitively declined adults. EEG entropy was differentially sensitive to task requirements across groups, with younger and older controls exhibiting greater control of encoding-related activity than older declined participants. Task sensitivity of frontal EEG during encoding predicted later retrieval, in line with previous evidence that cognitive decline is associated with reduced ability to self-initiate encoding-related processes.


Are dental color measuring devices CIE compliant?
This in vitro study evaluated the accordance of dental color measurement devices with a Commission Internationale de l'Eclairage (CIE)-compliant reference system, by comparing the CIE lightness, chroma, and hue (L*C*h°) color coordinates of ceramic samples. Four color measurement devices: Vita Easyshade® Advance (A); DeguDent Shadepilot™ (B); X-Rite® Shadevision (C); and Crystaleye Olympus (D), were compared with a CIE-compliant reference system by recording the L*C*h° color coordinates of ceramic samples matching the tooth colors of the Vita Linearguide 3D-Master®, under standardized test conditions. Differences between regression lines for the dental color measurement device data and regression lines for the CIE-compliant reference system data were evaluated. All devices offered high intraclass correlation coefficients (0.9771- 0.9999) for the L*C*h° color coordinates. The regression lines of the L* and C* coordinates for device A were steeper than those of the CIE-compliant reference system; the regression lines for devices B, C, and D were nearly parallel to those of the reference system, but with an offset. The regression lines of the h° coordinates for all devices were almost parallel to those of the reference system, with slopes near 1. Excluding the L* and h° coordinate measurements of device A, the measurements with the devices exhibited deviations from the reference system that were greater than those expected by chance (P < 0.0002). The dental color measurement devices assessed here offered excellent reproducibility, but showed significant deviations from the CIE-compliant reference system regarding the L*C*h° color coordinates.


Association between Stressful Life Events and Cognitive Disorders in Central Africa: Results from the EPIDEMCA Program.
Stressful life events (SLEs) are considered potential risk factors for cognitive disorders. Our objective was to investigate the association between SLEs and cognitive disorders among the elderly people in Central Africa. A population-based study was conducted in the Central African Republic (CAR) and the Republic of Congo (ROC). Participants aged ≥65 were interviewed using the Community Screening Interview for Dementia. Those who performed poorly were clinically assessed by neurologists. DSM-IV and Petersen criteria were required for a diagnosis of dementia or mild cognitive impairment (MCI), respectively. SLEs were assessed through 18 questions about events that occurred during childhood, adulthood and late-life. Sociodemographic, vascular and psychological factors were also documented. Multivariate multinomial logistic regression models were used to estimate the associations. MCI was positively associated with: the total number of SLEs (OR = 1.1, 95% CI: 1.0-1.2), the number of SLEs from the age of 65 (OR = 1.2, 95% CI: 1.0-1.3), the number of SLEs before the age of 16 among non-depressive participants (OR = 1.6, 95% CI: 1.2-2.2) and with a serious illness in a child experienced when the participant was aged 65 or more (OR = 2.8, 95% CI: 1.6-4.6). No association with dementia was observed. SLEs were positively associated with MCI but not dementia. More comprehensive studies are needed to further investigate this relationship.


3D-Printed seed planter and well array for high-throughput seed germination screening.
Seed germination is an important means of evaluating seed quality. In the present study, a well array for a seed germination experiment was designed and fabricated by 3D printing for the first time. Each hollow cone-shaped well can hold one seed, which not only prevented the seed from falling out of the well but also ensured that part of the seed was fully exposed to the sublayer of wet filter paper, allowing it to receive water for germination. Coupled with a 3D-printed seed planter, single seeds can be quickly placed into arrayed wells. Moreover, the number of sprouts could be automatically obtained from image analysis, which greatly improved the efficiency of the entire experiment. In summary, the high-throughput, easy-to-use seed germination well array that we have developed has been shown to have potential applications in botanical seed research and agricultural production.


Intermittent testicular torsion in the pediatric patient: sonographic indicators of a difficult diagnosis.
The purpose of this study was to determine whether intermittent testicular torsion, defined as the sudden onset of unilateral scrotal pain of short duration with spontaneous resolution, has sonographic indicators that can be used to establish the diagnosis. A retrospective search for pediatric patients presenting with suspected intermittent testicular torsion over a 2-year period was performed. The sonographic findings, medical records for clinical presentation, surgical outcomes, and comorbidities relevant to intermittent testicular torsion were reviewed. Analysis was performed on two separate categories for intermittent testicular torsion: total patient episodes (n = 19) and surgical cases (n = 10), with a total of 15 patients with 1.26 episodes per patient. Of the 19 episodes of intermittent testicular torsion, 63% (12) had either absent flow followed by normal to increased flow (n = 6, p = 0.18) or increased testicular flow (n = 6), 26% (5) had normal flow, and 10.5% (2) had decreased flow. Of the total episodes of intermittent testicular torsion, 79% (15) had a whirlpool sign or pseudomass, 10.5% (2) had neither a whirlpool sign or pseudomass but a "boggy" thickened cord, and 10.5% (2) had a normal appearance of the spermatic cord with no pseudomass. The whirlpool sign or an abnormal boggy cord and pseudomass formation below the twisted spermatic cord were both significant findings to support a diagnosis of intermittent testicular torsion. Although change from no perfusion to perfusion during the examination was not statistically significant in our cases, when present it led to the correct diagnosis in each case.


Distinct impacts of the 1997-98 and 2015-16 extreme El Niños on Japanese eel larval catch.
Extraordinarily poor recruitment of Japanese eels in East Asia has been generally reported during extreme El Niño years. However, the scenario failed to take place during the 2015-16 extreme event. In this study, we examined possible factors responsible for differing eel abundance in East Asia during the two strongest recent extreme El Niños, which occurred in 1997-98 and 2015-16. Numerical tracer experiments were carried out to determine why the impacts on eel catches seen in 1997-98 were not repeated in 2015-16. Among physical factors, two scenarios are likely responsible for extremely poor recruitment in East Asia: southward migration of the North Equatorial Current (NEC) or southward movement of eel spawning grounds. Comparing the latitudinal shift of NEC locations between the 1997-98 and 2015-16 El Niños, we conclude that NEC migration may be a factor, but is not chiefly responsible, for lower eel catches. Our findings pointed to southward movement of spawning grounds as the dominant factor. The northward movement of spawning grounds during 2015-16 meant that eel larvae were preferentially transported into the NEC-Kuroshio system, which resulted in a higher rate of recruitment success. The distinct evolution and dynamics of these two El Niño events led to different spawning ground locations, impacting eel abundance in East Asian countries.


Young athletes after ACL reconstruction with quadriceps strength asymmetry at the time of return-to-sport demonstrate decreased knee function 1 year later.
Quadriceps femoris (QF) strength deficits at return-to-sport (RTS) after ACL reconstruction (ACLR) contribute to decreased knee function at the same time point. However, the impact of QF strength at RTS on longitudinal function has not been examined. The purpose of this study was to test the hypothesis that young athletes after ACLR with QF strength asymmetry at RTS would demonstrate decreased knee-related function and lower proportions of functional recovery at 1 year post-RTS compared to young athletes following ACLR with nearly symmetric QF strength at RTS. Participants included 76 young athletes (74% female; mean age at RTS = 17.3 years) after primary, unilateral ACLR, cleared to RTS, and followed for 1 year after RTS. At the time of RTS, QF strength was quantified on an isokinetic dynamometer and a Limb Symmetry Index (LSI) was calculated [(involved/uninvolved) × 100%]. The cohort was subdivided into two groups based on RTS QF LSI: high quadriceps (HQ; LSI ≥ 90%; n = 36) and low quadriceps (LQ; LSI < 85%; n = 36). The cohort was followed for 1 year post-RTS, and knee-related function was assessed using the International Knee Documentation Committee subjective form (IKDC), the Knee Injury and Osteoarthritis Outcome Score (KOOS), and LSI of single-leg hop tests. Functional recovery at 1 year post-RTS was defined as KOOS scores above literature-reported cut-offs. While the HQ group demonstrated higher symmetry on all 1 year post-RTS hop tests, only the triple-hop test (p = 0.020) was found to be statistically different. Similarly, while the HQ group scored higher on all 1 year post-RTS self-reported knee function measures, only differences on the KOOS-Sport/Rec score (p = 0.039) and IKDC score (p = 0.011) were statistically different. Additionally, the HQ group demonstrated higher proportions of functional recovery at 1 year post-RTS than the LQ group on the KOOS-Symptoms (HQ: 88.9%, LQ: 69.4%; p = 0.040) and KOOS-Sport/Rec (HQ: 91.7%, LQ: 69.4%; p = 0.017). Young athletes after ACLR with QF strength asymmetry at RTS demonstrated decreased knee-related function and lower proportions of functional recovery at 1 year post-RTS. However, group differences did not exceed reported minimal clinically important difference values. Further study is warranted to understand factors that contribute to longitudinal knee function after ACLR. Clinicians should focus on restoring symmetric QF strength at RTS after ACLR, which may promote higher longitudinal knee function. Level II, Prospective cohort study.


Isolated Supravalvar Aortic Stenosis Without William's Syndrome.
Supravalvular aortic stenosis, characterized by narrowing of the ascending aorta above the valve, is the least common form of left ventricular outflow tract obstruction and is usually associated with William's syndrome. We present a case of a 27-year-old male with isolated supravalvar aortic stenosis (SVAS) presenting with heart failure. This case underscores the fact that in rare cases sporadic SVAS can occur in isolation without the classic findings of William's syndrome and highlighting the importance of integration of clinical and echocardiographic recognition for definitive management.


The consequences of climate-driven stop-over sites changes on migration schedules and fitness of Arctic geese.
1. How climatic changes affect migratory birds remains difficult to predict because birds use multiple sites in a highly interdependent manner. A better understanding of how conditions along the flyway affect migration and ultimately fitness is of paramount interest. 2. Therefore, we developed a stochastic dynamic model to generate spatially and temporally explicit predictions of stop-over site use. For each site, we varied energy expenditure, onset of spring, intake rate and day-to-day stochasticity independently. We parameterized the model for the migration of pink-footed goose Anser brachyrhynchus from its wintering grounds in Western Europe to its breeding grounds on Arctic Svalbard. 3. Model results suggested that the birds follow a risk-averse strategy by avoiding sites with comparatively high energy expenditure or stochasticity levels in favour of sites with highly predictable food supply and low expenditure. Furthermore, the onset of spring on the stop-over sites had the most pronounced effect on staging times while intake rates had surprisingly little effect. 4. Subsequently, using empirical data, we tested whether observed changes in the onset of spring along the flyway explain the observed changes in migration schedules of pink-footed geese from 1990 to 2004. Model predictions generally agreed well with empirically observed migration patterns, with geese leaving the wintering grounds earlier while considerably extending their staging times in Norway.


Sex hormones, insulin resistance, and diabetes mellitus among men with or at risk for HIV infection.
To examine the relationship of free testosterone (FT) and sex hormone-binding globulin (SHBG) with insulin resistance and diabetes mellitus (DM) in HIV disease. Cross-sectional analysis from 322 HIV-uninfected and 534 HIV-infected men in the Multicenter AIDS Cohort Study. : The main outcomes were DM and homeostasis model assessment-insulin resistance (HOMA-IR). DM was defined as fasting serum glucose ≥126 or self-reported DM and use of DM medications. HOMA-IR was calculated from fasting serum glucose and fasting insulin. Compared with HIV-uninfected men in our sample, HIV-infected men were younger, with lower body mass index, and more often black. HIV-infected men had lower FT (P < 0.001) and higher SHBG (P < 0.0001). The adjusted odds ratio for DM was 1.98 (95% confidence interval: 1.04 to 3.78); mean adjusted log HOMA-IR was 0.21 units higher in HIV-infected men (P < 0.0001). Log SHBG, but not log FT, was associated with DM (odds ratio = 0.44, 95% confidence interval: 0.25 to 0.80) in both groups. Log FT and log SHBG were inversely related to insulin resistance (P < 0.05 for both) independent of HIV. Compared with HIV-uninfected men, HIV-infected men had lower FT, higher SHBG, and more insulin resistance and DM. Lower FT and lower SHBG were associated with insulin resistance regardless of HIV serostatus. This suggests that sex hormones play a role in the pathogenesis of glucose abnormalities among HIV-infected men.


Neuropsychiatric disturbances in atypical parkinsonian disorders.
Multiple system atrophy (MSA), progressive supranuclear palsy (PSP), and corticobasal degeneration (CBD) are the most common atypical parkinsonisms. These disorders are characterized by varying combinations of autonomic, cerebellar and pyramidal system, and cognitive dysfunctions. In this paper, we reviewed the evidence available on the presence and type of neuropsychiatric disturbances in MSA, PSP, and CBD. A MedLine, Excerpta Medica, PsycLit, PsycInfo, and Index Medicus search was performed to identify all articles published on this topic between 1965 and 2018. Neuropsychiatric disturbances including depression, anxiety, agitation, and behavioral abnormalities have been frequently described in these disorders, with depression as the most frequent disturbance. MSA patients show a higher frequency of depressive disorders when compared to healthy controls. An increased frequency of anxiety disorders has also been reported in some patients, and no studies have investigated apathy. PSP patients may have depression, apathy, disinhibition, and to a lesser extent, anxiety and agitation. In CBD, neuropsychiatric disorders are similar to those present in PSP. Hallucinations and delusions are rarely reported in these disorders. Neuropsychiatric symptoms in MSA, PSP, and CBD do not appear to be related to the severity of motor dysfunction and are one of the main factors that determine a low quality of life. The results suggest that neuropsychiatric disturbances should always be assessed in patients with atypical parkinsonisms.


Bullying among medical and nonmedical students at a university in Eastern Saudi Arabia.
Many medical students, junior doctors, and other health-care professionals have been affected by the negative experience of bullying. Research is scarce on bullying experienced by medical and nonmedical students in Saudi Arabia unlike what is found in Western countries. It is unclear whether being a nonmedical student modifies the risk of being bullied. A cross-sectional study included 400 university students using convenient sampling. The sample comprised 295 students who responded and were stratified into medical (n = 176) and nonmedical (n = 119) groups. Statistical Package for the Social Sciences (SPSS) version 22.0 was used to analyze our data. Normality was measured using the Kolmogorov-Smirnov test. Statistical significance was tested using chi-square test for categorical variables, and t-test for continuous variables. Almost half of the respondents were found to have experienced some bullying, victimization, or other harassment during their medical education. The most common forms of bullying were verbal abuse and undue pressure to produce work (43.8%; n = 77). Nonmedical students experienced more bullying than medical students and were more likely to be female, single, and younger in age. The number of medical students subjected to sexual harassment (1.7%; n = 3) was higher than nonmedical students (0.8%; n = 1). Physical violence was more towards nonmedical (4.2%; n = 5) than medical students (1.1%, n = 2). The rates of bullying continue to be associated with anxiety and depression. Our data suggest similar bullying rates in the developed world but higher than previously reported in a Saudi study. Bullying or harassment affects both medical and nonmedical students and is associated with high levels of anxiety and depression.


Tailoring therapies--improving the management of early breast cancer: St Gallen International Expert Consensus on the Primary Therapy of Early Breast Cancer 2015.
The 14th St Gallen International Breast Cancer Conference (2015) reviewed substantial new evidence on locoregional and systemic therapies for early breast cancer. Further experience has supported the adequacy of tumor margins defined as 'no ink on invasive tumor or DCIS' and the safety of omitting axillary dissection in specific cohorts. Radiotherapy trials support irradiation of regional nodes in node-positive disease. Considering subdivisions within luminal disease, the Panel was more concerned with indications for the use of specific therapies, rather than surrogate identification of intrinsic subtypes as measured by multiparameter molecular tests. For the treatment of HER2-positive disease in patients with node-negative cancers up to 1 cm, the Panel endorsed a simplified regimen comprising paclitaxel and trastuzumab without anthracycline as adjuvant therapy. For premenopausal patients with endocrine responsive disease, the Panel endorsed the role of ovarian function suppression with either tamoxifen or exemestane for patients at higher risk. The Panel noted the value of an LHRH agonist given during chemotherapy for premenopausal women with ER-negative disease in protecting against premature ovarian failure and preserving fertility. The Panel noted increasing evidence for the prognostic value of commonly used multiparameter molecular markers, some of which also carried prognostic information for late relapse. The Panel noted that the results of such tests, where available, were frequently used to assist decisions about the inclusion of cytotoxic chemotherapy in the treatment of patients with luminal disease, but noted that threshold values had not been established for this purpose for any of these tests. Multiparameter molecular assays are expensive and therefore unavailable in much of the world. The majority of new breast cancer cases and breast cancer deaths now occur in less developed regions of the world. In these areas, less expensive pathology tests may provide valuable information. The Panel recommendations on treatment are not intended to apply to all patients, but rather to establish norms appropriate for the majority. Again, economic considerations may require that less expensive and only marginally less effective therapies may be necessary in less resourced areas. Panel recommendations do not imply unanimous agreement among Panel members. Indeed, very few of the 200 questions received 100% agreement from the Panel. In the text below, wording is intended to convey the strength of Panel support for each recommendation, while details of Panel voting on each question are available in supplementary Appendix S2, available at Annals of Oncology online.


Aberrant accumulation of undifferentiated myeloid cells in the adipose tissue of CCR2-deficient mice delays improvements in insulin sensitivity.
Mice with CCR2 deficiency are protected from insulin resistance but only after long periods of high-fat diet (HFD) feeding, despite the virtual absence of circulating inflammatory monocytes. We performed a time course study in mice with hematopoietic and global CCR2 deficiency to determine adipose tissue-specific mechanisms for the delayed impact of CCR2 deficiency on insulin resistance. Mice with global or hematopoietic CCR2 deficiency (CCR2(-/-) and BM-CCR2(-/-), respectively) and wild-type controls (CCR2(+/+) and BM-CCR2(+/+), respectively) were placed on an HFD for 6, 12, and 20 weeks. Adipose tissue myeloid populations, degree of inflammation, glucose tolerance, and insulin sensitivity were assessed. Flow cytometry analysis showed that two different populations of F4/80(+) myeloid cells (CD11b(lo)F4/80(lo) and CD11b(hi)F4/80(hi)) accumulated in the adipose tissue of CCR2(-/-) and BM-CCR2(-/-) mice after 6 and 12 weeks of HFD feeding, whereas only the CD11b(hi)F4/80(hi) population was detected in the CCR2(+/+) and BM-CCR2(+/+) controls. After 20 weeks of HFD feeding, the CD11b(lo)F4/80(lo) cells were no longer present in the adipose tissue of CCR2(-/-) mice, and only then were improvements in adipose tissue inflammation detected. Gene expression and histological analysis of the CD11b(lo)F4/80(lo) cells indicated that they are a unique undifferentiated monocytic inflammatory population. The CD11b(lo)F4/80(lo) cells are transiently found in wild-type mice, but CCR2 deficiency leads to the aberrant accumulation of these cells in adipose tissue. The discovery of this novel adipose tissue monocytic cell population provides advances toward understanding the pleiotropic role of CCR2 in monocyte/macrophage accumulation and regulation of adipose tissue inflammation.


RNA:DNA hybrids are a novel molecular pattern sensed by TLR9.
The sensing of nucleic acids by receptors of the innate immune system is a key component of antimicrobial immunity. RNA:DNA hybrids, as essential intracellular replication intermediates generated during infection, could therefore represent a class of previously uncharacterised pathogen-associated molecular patterns sensed by pattern recognition receptors. Here we establish that RNA:DNA hybrids containing viral-derived sequences efficiently induce pro-inflammatory cytokine and antiviral type I interferon production in dendritic cells. We demonstrate that MyD88-dependent signalling is essential for this cytokine response and identify TLR9 as a specific sensor of RNA:DNA hybrids. Hybrids therefore represent a novel molecular pattern sensed by the innate immune system and so could play an important role in host response to viruses and the pathogenesis of autoimmune disease.


Single-step, self-etch adhesives behave as permeable membranes after polymerization. Part III. Evidence from fluid conductance and artificial caries inhibition.
To test the hypothesis that formation of inhibition zones by bonded restorations in artificially-induced carious dentin lesions is related to the permeability of the self-etch adhesives. Fluid conductance of four single-step self-etch adhesives (Prompt L-Pop, Etch&Prime 3.0, One-Up Bond F and Reactmer Bond), and a control two-step self-etch adhesive (UniFil Bond) were measured after sound dentin surfaces were:(1) acid-etched; (2) polished to produce smear layers; (3) bonded with the adhesive; each at 20 cm hydrostatic pressure (hp). Osmotic fluid conductance was also determined for the bonded dentin after immersion in 4.8 M CaCl2 at 0 cm hp. For artificial caries inhibition, a non-fluoride releasing (NFR) two-step self-etch adhesive (UniFil Bond) and a fluoride-releasing (FR) single-step adhesive (Reactmer Bond) were used in combination with a NFR composite (Metafil CX) or a FR restorative material (Reactmer Paste). Artificial caries were induced in these restorations, from which 120 +/- 10 microm thick sections and the effect of caries inhibition were quantitatively assessed with polarizing light microscopy and image analysis. Fluid conductance after bonding with the single-step adhesives Prompt L-Pop and Etch&Prime 3.0 were not significantly different from those recorded from smear layer-covered dentin. Although better seals were achieved with One-Up Bond F and Reactmer Bond, fluid conductance at 20 cm hp was significantly higher than the two-step self-etch adhesive UniFil Bond. Osmotic conductance at 0 cm hp were not significantly different from normal fluid conductance at 20 cm hp. The sizes of artificial carious lesions in UniFil Bond were significantly higher irrespective of whether a FR or NFR composite was used. Significant reduction in lesion size occurred in Reactmer Bond with the use of a NFR composite, and even more so with the use of a FR composite.


Dietary polyphenols and mechanisms of osteoarthritis.
Osteoarthritis is a condition caused in part by injury, loss of cartilage structure and function, and an imbalance in inflammatory and anti-inflammatory pathways. It primarily affects the articular cartilage and subchondral bone of synovial joints and results in joint failure, leading to pain upon weight bearing including walking and standing. There is no cure for osteoarthritis, as it is very difficult to restore the cartilage once it is destroyed. The goals of treatment are to relieve pain, maintain or improve joint mobility, increase the strength of the joints and minimize the disabling effects of the disease. Recent studies have shown an association between dietary polyphenols and the prevention of osteoarthritis-related musculoskeletal inflammation. This review discusses the effects of commonly consumed polyphenols, including curcumin, epigallocatechin gallate and green tea extract, resveratrol, nobiletin and citrus fruits, pomegranate, as well as genistein and soy protein, on osteoarthritis with an emphasis on molecular antiosteoarthritic mechanisms.


Monitoring of hydrogen sulfide via substrate-integrated hollow waveguide mid-infrared sensors in real-time.
Hydrogen sulfide is a highly corrosive, harmful, and toxic gas produced under anaerobic conditions within industrial processes or in natural environments, and plays an important role in the sulfur cycle. According to the U.S. Occupational Safety and Health Administration (OSHA), the permissible exposure limit (during 8 hours) is 10 ppm. Concentrations of 20 ppm are the threshold for critical health issues. In workplace environments with human subjects frequently exposed to H2S, e.g., during petroleum extraction and refining, real-time monitoring of exposure levels is mandatory. Sensors based on electrochemical measurement principles, semiconducting metal-oxides, taking advantage of their optical properties, have been described for H2S monitoring. However, extended response times, limited selectivity, and bulkiness of the instrumentation are common disadvantages of the sensing techniques reported to date. Here, we describe for the first time usage of a new generation of compact gas cells, i.e., so-called substrate-integrated hollow waveguides (iHWGs), combined with a compact Fourier transform infrared (FTIR) spectrometer for advanced gas sensing of H2S. The principle of detection is based on the immediate UV-assisted conversion of the rather weak IR-absorber H2S into much more pronounced and distinctively responding SO2. A calibration was established in the range of 10-100 ppm with a limit of detection (LOD) at 3 ppm, which is suitable for occupational health monitoring purposes. The developed sensing scheme provides an analytical response time of less than 60 seconds. Considering the substantial potential for miniaturization using e.g., a dedicated quantum cascade laser (QCL) in lieu of the FTIR spectrometer, the developed sensing approach may be evolved into a hand-held instrument, which may be tailored to a variety of applications ranging from environmental monitoring to workplace safety surveillance, process analysis and clinical diagnostics, e.g., breath analysis.


European Organisation for Research and Treatment of Cancer Soft Tissue and Bone Sarcoma Group Experience with Advanced/Metastatic Epithelioid Sarcoma Patients Treated in Prospective Trials: Clinical Profile and Response to Systemic Therapy.
Epithelioid sarcoma is a soft tissue sarcoma associated with a high rate of local recurrence after wide resection and high incidence of distant metastasis. Little is known about the clinical course and response to systemic treatments in epithelioid sarcoma patients. We carried out a retrospective analysis of clinical data from epithelioid sarcoma patients to provide a reference for the design of future epithelioid sarcoma-specific studies. Data from patients with epithelioid sarcoma entered in prospective multi-sarcoma phase II/III trials were pooled: EORTC trial 62012 (doxorubicin versus doxorubicin/ifosfamide), 62043 (pazopanib), 62072 (pazopanib versus placebo) and 62091 (doxorubicin versus trabectedin). Patients had either a local or a centrally confirmed diagnosis of epithelioid sarcoma, had inoperable/metastatic disease at study entry and were eligible for the according trial. Response was assessed according to RECIST 1.1. Progression-free survival (PFS) and overall survival were calculated from date of entry. Among 976 patients with advanced sarcomas, 27 epithelioid sarcoma patients (2.8%) were eligible for the analysis (17 men, median age at diagnosis 50 years, range 19-72). Eighteen (66.7%) received chemotherapy as first-line treatment (five doxorubicin, eight doxorubicin/ifosfamide, two pazopanib, three trabectedin) and nine (33.3%) received pazopanib as second line or later. The primary tumour was located in the lower extremity (n = 8; 29.6%), upper extremity (n = 5; 18.5%), retro/intra-abdominal (n = 4; 14.8%) and in other locations (n = 10; 37.0%). At entry, metastases were mainly found in lung (n = 17; 63%), lymph nodes (n = 9; 33.3%), bone (n = 8; 29.6%) and soft tissue (n = 7; 25.9%). The best response for first-line patients was four partial responses (22.2%), 10 stable disease (55.6%) and four progressive disease (22.2%). In subsequent lines, pazopanib achieved one partial response (11.1%), four stable disease (44.4%) and four progressive disease (44.4%). All patients but one progressed on treatment. The median PFS and overall survival were 3.8 (95% confidence interval 2.2-4.8) and 10.8 months (95% confidence interval 8.1-21.3), respectively. Five patients were still alive at the time of the according trial analysis. With all limitations of such a rare disease and small data set, objective response and survival outcomes are similar in epithelioid sarcoma to non-selected sarcoma populations. The clinical testing of novel systemic treatments for epithelioid sarcoma remains an unmet medical need and a high priority.