Torsion angle preference and energetics of small-molecule ligands bound to proteins.
Small organic molecules can assume conformations in the protein-bound state that are significantly different from those in solution. We have analyzed the conformations of 21 common torsion motifs of small molecules extracted from crystal structures of protein-ligand complexes and compared them with their torsion potentials calculated by an ab initio DFT method. We find a good correlation between the potential energy of the torsion motifs and their conformational distribution in the protein-bound state: The most probable conformations of the torsion motifs agree well with the calculated global energy minima, and the lowest torsion-energy state becomes increasingly dominant as the torsion barrier height increases. The torsion motifs can be divided into 3 groups based on torsion barrier heights: high (>4 kcal/mol), medium (2-4 kcal/mol), and low (<2 kcal/mol). The calculated torsion energy profiles are predictive for the most preferred bound conformation for the high and medium barrier groups, the latter group common in druglike molecules. In the high-barrier group of druglike ligands, >95% of conformational torsions occur in the energy region <4 kcal/mol. The conformations of the torsion motifs in the protein-bound state can be modeled by a Boltzmann distribution with a temperature factor much higher than room temperature. This high-temperature factor, derived by fitting the theoretical model to the experimentally observed conformation occurrence of torsions, can be interpreted as the perturbation that proteins inflict on the conformation of the bound ligand. Using this model, it is calculated that the average strain energy of a torsion motif in ligands bound to proteins is approximately 0.6 kcal/mol, a result which can be related to the lower binding efficiency of larger ligands with more rotatable bonds. The above results indicate that torsion potentials play an important role in dictating ligand conformations in both the free and the bound states.


Comparison of Three Information Sources for Smoking Information in Electronic Health Records.
The primary aim was to compare independent and joint performance of retrieving smoking status through different sources, including narrative text processed by natural language processing (NLP), patient-provided information (PPI), and diagnosis codes (ie, International Classification of Diseases, Ninth Revision [ICD-9]). We also compared the performance of retrieving smoking strength information (ie, heavy/light smoker) from narrative text and PPI. Our study leveraged an existing lung cancer cohort for smoking status, amount, and strength information, which was manually chart-reviewed. On the NLP side, smoking-related electronic medical record (EMR) data were retrieved first. A pattern-based smoking information extraction module was then implemented to extract smoking-related information. After that, heuristic rules were used to obtain smoking status-related information. Smoking information was also obtained from structured data sources based on diagnosis codes and PPI. Sensitivity, specificity, and accuracy were measured using patients with coverage (ie, the proportion of patients whose smoking status/strength can be effectively determined). NLP alone has the best overall performance for smoking status extraction (patient coverage: 0.88; sensitivity: 0.97; specificity: 0.70; accuracy: 0.88); combining PPI with NLP further improved patient coverage to 0.96. ICD-9 does not provide additional improvement to NLP and its combination with PPI. For smoking strength, combining NLP with PPI has slight improvement over NLP alone. These findings suggest that narrative text could serve as a more reliable and comprehensive source for obtaining smoking-related information than structured data sources. PPI, the readily available structured data, could be used as a complementary source for more comprehensive patient coverage.


The Internet as a new tool in the rehabilitation process of patients--education in focus.
In the article we deal with the rehabilitation of patients using information technology, especially Internet support. We concentrate on two main areas in the IT support of rehabilitation: one of them is the support for individual therapy, the other one is providing patients with information, which is the basic step in emphasising individual responsibility. In the development of rehabilitation programmes, the knowledge of the IT professional and the therapist, in the IT support of web guidance, medical expertise plays the primary role. The degree of assistance involved in the rehabilitation process depends on the IT knowledge of medical (general practitioner, nursing staff) professionals as well. The necessary knowledge required in healing and development processes is imparted to professionals by a special (full-time) university training. It was a huge challenge for us to teach web-based information organisation skills to doctors and nurses, and it is also a complex task to put forward such an IT viewpoint to information specialists in order to create the foundations of the cooperation between IT and healthcare professionals.


Live and let die: a REM complex promotes fertilization through synergid cell death in Arabidopsis.
Fertilization in flowering plants requires a complex series of coordinated events involving interaction between the male and female gametophyte. We report here molecular data on one of the key events underpinning this process - the death of the receptive synergid cell and the coincident bursting of the pollen tube inside the ovule to release the sperm. We show that two REM transcription factors, VALKYRIE (VAL) and VERDANDI (VDD), both targets of the ovule identity MADS-box complex SEEDSTICK-SEPALLATA3, interact to control the death of the receptive synergid cell. In vdd-1/+ mutants and VAL_RNAi lines, we find that GAMETOPHYTIC FACTOR 2 (GFA2), which is required for synergid degeneration, is downregulated, whereas expression of FERONIA (FER) and MYB98, which are necessary for pollen tube attraction and perception, remain unaffected. We also demonstrate that the vdd-1/+ phenotype can be rescued by expressing VDD or GFA2 in the synergid cells. Taken together, our findings reveal that the death of the receptive synergid cell is essential for maintenance of the following generations, and that a complex comprising VDD and VAL regulates this event.


Waist-to-Height Ratio, Aerobic Fitness, and Cardiometabolic Risk in Hispanic Children From Low-Income U.S. Schools.
The purpose of this study was to examine the relationships among waist-to-height ratio (WHtR), aerobic fitness, and cardio-metabolic risk factors in Hispanic children from low-income U.S. schools. Participants were 198 Hispanic children from low-income schools (Mean age = 10.3 ± 0.5 years; 119 girls, 79 boys). Waist circumference, height, and cardio-metabolic blood markers were collected in a fasted state. Estimated VO2 Peak scores were also collected. Multilevel generalized mixed effects models were employed to examine the independent effect of WHtR and aerobic fitness classification on a child meeting recommended levels for each cardio-metabolic blood marker. A child having a WHtR < 0.5 related to meeting recommended levels for HDL cholesterol (OR = 3.25, p < .01), triglycerides (OR = 2.94, p < .01), glucose (OR = 3.42, p < .01), and related to a lower continuous Mean Arterial Pressure (MAP) score (β = -8.5 mmHg, p < .01). Aerobic fitness classification only independently related to meeting recommended levels for HDL cholesterol (OR = 2.94, p = .010). Having a WHtR < 0.5 independently associated with favorable cardio-metabolic blood markers and thus serves as an effective screening tool for cardio-metabolic risk in Hispanic children from low-income schools.


A complete expression profile of matrix-degrading metalloproteinases in Dupuytren's disease.
Dupuytren's disease (DD) is a common fibrotic condition of the palmar fascia, leading to deposition of collagen-rich cords and finger contractions. The metzincin superfamily contains key enzymes in the turnover of collagen and other extracellular matrix macromolecules. A number of broad-spectrum matrix metalloproteinase inhibitors, used in cancer clinical trials, caused side effects of DD-like contractures. We tested the hypothesis that changes in the expression of specific metalloproteinases underlie or contribute to the fibrosis and contracture seen in DD. We collected tissue from patients with DD and used normal palmar fascia as a control. We profiled the expression of the entire matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinases (TIMP), and a disintegrin and metalloproteinase domain with thrombospondin motif (ADAMTS) gene families in these tissues using real-time reverse-transcription polymerase chain reaction. A number of metalloproteinases and inhibitors are regulated in DD. The expression of 3 key collagenases, MMP1, MMP13, and MMP14 is increased significantly in the DD nodule, as is the expression of the collagen biosynthetic enzyme ADAMTS14. The expression of MMP7, an enzyme with broad substrate specificity, is increased in the DD nodule and remains equally expressed in the DD cord. TIMP1 expression is increased significantly in DD nodule compared with normal palmar fascia. This study measured the expression of all MMP, ADAMTS, and TIMP genes in DD. Contraction and fibrosis may result from: (1) increased collagen biosynthesis mediated by increased ADAMTS-14; (2) an increased level of TIMP-1 blocking MMP-1- and MMP-13-mediated collagenolysis; and (3) contraction enabled by MMP-14-mediated pericellular collagenolysis (and potentially MMP-7), which may escape inhibition by TIMP-1. The complete expression profile will provide a knowledge-based approach to novel therapeutics targeting these genes.


Co-creation of information leaflets to meet the support needs of people living with complex regional pain syndrome (CRPS) through innovative use of wiki technology.
People living with complex regional pain syndrome (CRPS) experience frustration with the lack of knowledge and understanding of CRPS as a pain condition. We report on our attempt to address this issue. People living with CRPS taking part in a larger study were invited to co-construct a CRPS wiki page that addressed the areas in which they had experienced the most difficulty. A blank wiki page was set up for participants to populate with issues they felt needed to be raised and addressed. Participants failed to engage with the wiki technology. We modified our procedure and completed an inductive analysis of a sister-forum which participants were using as part of the larger study. Six issues of importance were identified. We used the discussion forum threads to populate the themes. Due to a continued lack of engagement with the wiki technology, the team decided to create a suite of leaflets which were piloted with delegates at a CRPS patient conference. Future work should be mindful of the extent to which patients are able and willing to share their experiences through such technology. Striking the balance between patient-endorsed and researcher-driven co-creation of such material is imperative.


The FgfrL1 receptor is required for development of slow muscle fibers.
FgfrL1, which interacts with Fgf ligands and heparin, is a member of the fibroblast growth factor receptor (Fgfr) family. FgfrL1-deficient mice show two significant alterations when compared to wildtype mice: They die at birth due to a malformed diaphragm and they lack metanephric kidneys. Utilizing gene arrays, qPCR and in situ hybridization we show here that the diaphragm of FgfrL1 knockout animals lacks any slow muscle fibers at E18.5 as indicated by the absence of slow fiber markers Myh7, Myl2 and Myl3. Similar lesions are also found in other skeletal muscles that contain a high proportion of slow fibers at birth, such as the extraocular muscles. In contrast to the slow fibers, fast fibers do not appear to be affected as shown by expression of fast fiber markers Myh3, Myh8, Myl1 and MylPF. At early developmental stages (E10.5, E15.5), FgfrL1-deficient animals express slow fiber genes at normal levels. The loss of slow fibers cannot be attributed to the lack of kidneys, since Wnt4 knockout mice, which also lack metanephric kidneys, show normal expression of Myh7, Myl2 and Myl3. Thus, FgfrL1 is specifically required for embryonic development of slow muscle fibers.


Water–methanol separation with carbon nanotubes and electric fields.
Methanol is used in various applications, such as fuel for transportation vehicles, fuel cells, and in chemical industrial processes. Conventionally, separation of methanol from aqueous solution is by distillation. However, this method consumes a large amount of energy; hence development of a new method is needed. In this work, molecular dynamics simulations are performed to investigate the effect of an electric field on water–methanol separation by carbon nanotubes (CNTs) with diameters of 0.81 to 4.07 nm. Without an electric field, methanol molecules fill the CNTs in preference to water molecules. The preference of methanol to occupy the CNTs over water results in a separation effect. This separation effect is strong for small CNT diameters and significantly decreases with increasing diameter. In contrast, under an electric field, water molecules strongly prefer to occupy the CNTs over methanol molecules, resulting in a separation effect for water. More interestingly, the separation effect for water does not decrease with increasing CNT diameter. Formation of water structures in CNTs induced by an electric field has an important role in the separation of water from methanol.


Enacted stigma experiences and protective factors are strongly associated with mental health outcomes of transgender people in Aotearoa/New Zealand.
International evidence has found large mental health inequities among transgender people and demonstrates that mental health outcomes are associated with enacted stigma experiences and protective factors. This study aimed to examine the extent of associations of enacted stigma experiences specific to transgender people alongside protective factors with mental health of transgender people in Aotearoa/New Zealand. The 2018 Counting Ourselves survey was a nationwide community-based study of transgender people (N = 1178, Mage = 29.5) living in Aotearoa/New Zealand. The survey assessed a wide range of gender minority stress experiences and protective factors that comprised primary (support from friends and family) and secondary social ties (neighborhood and transgender community belongingness). We calculated the predicted probabilities that transgender people exhibit very high psychological distress level, non-suicidal self-injury, and suicidal risks with different combinations and exposure profiles of enacted stigma and protective factors. Our findings demonstrated that enacted stigma was associated with negative mental health, and support of friends and family was linked to better outcomes across all mental health measures. Beyond primary social ties, sense of belongingness to neighborhood and transgender communities were linked to reduced odds of psychological distress and suicidal ideation. For those scoring high on enacted stigma and low on protective factors, our model revealed a 25% probability of attempting suicide in the last year compared to 3% for those scoring low on enacted stigma and high on protective factors. Echoing previous findings, this study demonstrates that transgender people across Aotearoa/New Zealand are less likely to manifest life-threatening mental health outcomes if they experience low levels of enacted stigma and high levels of access to protective factors. Our findings suggest a need to address the enacted stigma that transgender people face across interpersonal and structural settings, and also to enhance social supports that are gender affirmative for this population.


Restriction of oral intake of water for aspiration lung disease in children.
Primary aspiration of food and fluid is commonly seen in children with feeding and swallowing difficulties associated with a range of diseases and complex medical conditions. Respiratory sequelae and pneumonia are known to be associated with primary aspiration of ingested material, however causality between primary aspiration of specific food and fluid types and pulmonary effects in children is yet to be established in controlled trials. The relative pulmonary morbidity of aspiration of ingested food and fluid materials versus other causes of respiratory disease such as viral and bacterial causes, secondary aspiration of gastrointestinal contents and predisposing lung conditions such as chronic neonatal lung disease in a developing immune system is also unclear. Current management decisions for children who aspirate have to optimise oral nutrition and hydration, while reducing the risk of aspiration to preserve pulmonary integrity. This generally includes restricting aspirated food or fluids and providing texture-modified diets and thickened fluids. Young children frequently refuse thickened fluids providing a management dilemma for both families and health professionals. Our objective was to evaluate the efficacy of restriction of oral water ingestion on the pulmonary status of children with thin fluid aspiration demonstrated on a modified barium swallow study. The Cochrane Central Register of Controlled Trials (CENTRAL), the Cochrane Airways Collaborative Review Group Specialised Register, MEDLINE, EMBASE and CINAHL databases were searched by the Cochrane Airways Group. The latest search was performed in May 2102. All randomised controlled trials comparing restriction of oral intake of water with unlimited oral water ingestion were eligible to be included. Results of searches were reviewed against a pre-determined criteria for inclusion. No eligible trials were identified for a paediatric population and thus no data were available for analysis. One trial in an adult population was identified and reported. No randomised controlled trials examining the efficacy of restriction of oral intake of water in the management of children with thin fluid aspiration were found. In a single study in an adult population with stroke, no significant differences were seen between a control group of oral water restriction and the experimental group of unlimited oral water ingestion on outcomes such as pneumonia, total oral fluid intake and dehydration. There are no trials that have adequately evaluated the pulmonary effects of allowing or restricting oral water ingestion in children known to have primary aspiration of thin fluids. Thus, there is currently an absence of evidence to support a strict approach of full restriction of oral intake of water or support a more liberal approach of allowing oral water ingestion in children with primary aspiration of thin fluids.


Homocysteine alters the osteoprotegerin/RANKL system in the osteoblast to promote bone loss: pivotal role of the redox regulator forkhead O1.
In this study we determined the molecular mechanisms of how homocysteine differentially affects receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) synthesis in the bone. The results showed that oxidative stress induced by homocysteine deranges insulin-sensitive FOXO1 and MAP kinase signaling cascades to decrease OPG and increase RANKL synthesis in osteoblast cultures. We observed that downregulation of insulin/FOXO1 and p38 MAP kinase signaling mechanisms due to phosphorylation of protein phosphatase 2A (PP2A) was the key event that inhibited OPG synthesis in homocysteine-treated osteoblast cultures. siRNA knockdown experiments confirmed that FOXO1 is integral to OPG and p38 synthesis. Conversely homocysteine increased RANKL synthesis in osteoblasts through c-Jun/JNK MAP kinase signaling mechanisms independent of FOXO1. In the rat bone milieu, high-methionine diet-induced hyperhomocysteinemia lowered FOXO1 and OPG expression and increased synthesis of proresorptive and inflammatory cytokines such as RANKL, M-CSF, IL-1α, IL-1β, G-CSF, GM-CSF, MIP-1α, IFN-γ, IL-17, and TNF-α. Such pathophysiological conditions were exacerbated by ovariectomy. Lowering the serum homocysteine level by a simultaneous supplementation with N-acetylcysteine improved OPG and FOXO1 expression and partially antagonized RANKL and proresorptive cytokine synthesis in the bone milieu. These results emphasize that hyperhomocysteinemia alters the redox regulatory mechanism in the osteoblast by activating PP2A and deranging FOXO1 and MAPK signaling cascades, eventually shifting the OPG:RANKL ratio toward increased osteoclast activity and decreased bone quality.


Competing autocrine pathways involving alternative neuropilin-1 ligands regulate chemotaxis of carcinoma cells.
Neuropilin-1 (NP1), in conjunction with plexins, promotes axon repulsion by binding to semaphorin 3A (SEMA3A). Although NP1 is expressed in carcinoma cells, its functions have remained elusive, and neither SEMA3A nor plexin expression has been explored in cancer. Here we provide evidence that breast carcinoma cells support an autocrine pathway involving SEMA3A, plexin-A1, and NP1 that impedes their ability to chemotax. Reducing SEMA3A or NP1 expression by RNA interference or inhibiting plexin-A1 signaling enhanced migration. Conversely, expression of constitutively active plexin-A1 impaired chemotaxis. The paradox of how breast carcinoma cells expressing these endogenous chemotaxis inhibitors are able to migrate is explained by their expression of vascular endothelial growth factor (VEGF), a NP1 ligand that competes with SEMA3A for receptor binding. Finally, we establish that the ratio of endogenous VEGF and SEMA3A concentrations in carcinoma cells determines their chemotactic rate. Our findings lead to the surprising conclusion that opposing autocrine loops involving NP1 regulate the chemotaxis of breast carcinoma cells. Moreover, our data indicate a novel autocrine function for VEGF in chemotaxis.


Interleukin-15-activated natural killer cells kill autologous osteoclasts via LFA-1, DNAM-1 and TRAIL, and inhibit osteoclast-mediated bone erosion in vitro.
Osteoclasts reside on bone and are the main bone resorbing cells playing an important role in bone homeostasis, while natural killer (NK) cells are bone-marrow-derived cells known to play a crucial role in immune defence against viral infections. Although mature NK cells traffic through bone marrow as well as to inflammatory sites associated with enhanced bone erosion, including the joints of patients with rheumatoid arthritis, little is known about the impact NK cells may have on mature osteoclasts and bone erosion. We studied the interaction between human NK cells and autologous monocyte-derived osteoclasts from healthy donors in vitro. We show that osteoclasts express numerous ligands for receptors present on activated NK cells. Co-culture experiments revealed that interleukin-15-activated, but not resting, NK cells trigger osteoclast apoptosis in a dose-dependent manner, resulting in drastically decreased bone erosion. Suppression of bone erosion requires contact between NK cells and osteoclasts, but soluble factors also play a minor role. Antibodies masking leucocyte function-associated antigen-1, DNAX accessory molecule-1 or tumour necrosis factor-related apoptosis-inducing ligand enhance osteoclast survival when co-cultured with activated NK cells and restore the capacity of osteoclasts to erode bone. These results suggest that interleukin-15-activated NK cells may directly affect bone erosion under physiological and pathological conditions.


Rapid rates of newly synthesized mitochondrial protein degradation are significantly affected by the generation of mitochondrial free radicals.
Exposure of biological material to high levels of free radicals causes extensive cellular damage. Reactive oxygen species (ROS) generated by mitochondria have been associated with a variety of diseases and aging. We investigated the effect of low-level mitochondrial ROS production on newly synthesized mitochondrial proteins which are potentially vulnerable to mitochondrial ROS due to their location and unfolded state. We show that elevated mitochondrial ROS increases the degradation of newly synthesized mitochondrial proteins with some proteins more sensitive than others. In the long term reduced assembly of mitochondrial complexes would affect mitochondrial function and may trigger a vicious cycle of mitochondrial ROS production.


Reduced IL-12 production by monocytes upon ultraviolet-B irradiation selectively limits activation of T helper-1 cells.
The capacity of APC to stimulate the proliferation of human peripheral blood T cells decreases upon ultraviolet-B (UVB) irradiation. The aim of this study was to investigate whether all T cell subsets are equally sensitive to this reduced APC function. Established human Th1, Th2, and Th0 clones were stimulated with monocytes in a soluble CD3 mAb-mediated assay that is dependent on the presence of APC. Monocytes were exposed to low nonlethal doses of UVB radiation before coculture with T cells. UVB irradiation inhibited the capacity of monocytes to stimulate the proliferation and IFN-gamma production of Th1 cells in a dose-related fashion. In contrast, UVB-treated monocytes induced normal proliferation and IL-4 production in Th2 cells. Stimulation of Th0 cell proliferation by UVB-irradiated monocytes was normal, but a preferential suppression of IFN-gamma production was observed, thus leading to a more Th2-like cytokine response. The loss of Th1 proliferation upon stimulation with UVB-irradiated monocytes could be overcome by rIL-2; however, IFN-gamma production remained suppressed. IFN-gamma production could be completely restored by rIL-12, whereas the addition of IL-1 beta, TNF-alpha, or indomethacin had no such effect, nor did the addition of mAb to CD28, added to compensate for the reduced B7 expression of UVB-irradiated monocytes. Monocytes exposed to UVB radiation exhibited reduced expression of mRNA for the IL-1 2 subunits p35 and p40 and suppressed production of the IL-12 p70 protein. Our results thus indicate that UVB irradiation of APC selectively impairs Th1-like responses, a phenomenon caused by the UVB-induced suppression of monocyte IL-12 production.


Mechanisms of Targeting the MDM2-p53-FOXM1 Axis in Well-Differentiated Intestinal Neuroendocrine Tumors.
The tumor suppressor p53 is rarely mutated in gastroenteropancreatic neuroendocrine neoplasms (GEP-NEN) but they frequently show a strong expression of negative regulators of p53, rendering these tumors excellent targets for a p53 recovery therapy. Therefore, we analyzed the mechanisms of a p53 recovery therapy on intestinal neuroendocrine tumors in vitro and in vivo. By Western blot and immunohistochemistry, we found that in GEP-NEN biopsy material overexpression of MDM2 was present in intestinal NEN. Therefore, we analyzed the effect of a small-molecule inhibitor, nutlin-3a, in p53 wild-type and mutant GEP-NEN cell lines by proliferation assay, flow cytometry, immunofluorescence, Western blot, and by multiplex gene expression analysis. Finally, we analyzed the antitumor effect of nutlin-3a in a xenograft mouse model in vivo. During the study, the tumor volume was determined. The midgut wild-type cell line KRJ-I responded to the treatment with cell cycle arrest and apoptosis. By gene expression analysis, we could demonstrate that nutlins reactivated an antiproliferative p53 response. KRJ-I-derived xenograft tumors showed a significantly decreased tumor growth upon treatment with nutlin-3a in vivo. Furthermore, our data suggest that MDM2 also influences the expression of the oncogene FOXM1 in a p53-independent manner. Subsequently, a combined treatment of nutlin-3a and cisplatin (as chemoresistance model) resulted in synergistically enhanced antiproliferative effects. In summary, MDM2 overexpression is a frequent event in p53 wild-type intestinal neuroendocrine neoplasms and therefore recovery of a p53 response might be a novel personalized treatment approach in these tumors.


Prioritising references for systematic reviews with RobotAnalyst: A user study.
Screening references is a time-consuming step necessary for systematic reviews and guideline development. Previous studies have shown that human effort can be reduced by using machine learning software to prioritise large reference collections such that most of the relevant references are identified before screening is completed. We describe and evaluate RobotAnalyst, a Web-based software system that combines text-mining and machine learning algorithms for organising references by their content and actively prioritising them based on a relevancy classification model trained and updated throughout the process. We report an evaluation over 22 reference collections (most are related to public health topics) screened using RobotAnalyst with a total of 43 610 abstract-level decisions. The number of references that needed to be screened to identify 95% of the abstract-level inclusions for the evidence review was reduced on 19 of the 22 collections. Significant gains over random sampling were achieved for all reviews conducted with active prioritisation, as compared with only two of five when prioritisation was not used. RobotAnalyst's descriptive clustering and topic modelling functionalities were also evaluated by public health analysts. Descriptive clustering provided more coherent organisation than topic modelling, and the content of the clusters was apparent to the users across a varying number of clusters. This is the first large-scale study using technology-assisted screening to perform new reviews, and the positive results provide empirical evidence that RobotAnalyst can accelerate the identification of relevant studies. The results also highlight the issue of user complacency and the need for a stopping criterion to realise the work savings.


Engineering aggregation resistance in IgG by two independent mechanisms: lessons from comparison of Pichia pastoris and mammalian cell expression.
Aggregation is an important concern for therapeutic antibodies, since it can lead to reduced bioactivity and increase the risk of immunogenicity. In our analysis of immunoglobulin G (IgG) molecules of identical amino acid sequence but produced either in mammalian cells (HEK293) or in the yeast Pichia pastoris (PP), dramatic differences in their aggregation susceptibilities were encountered. The antibodies produced in Pichia were much more resistant to aggregation under many conditions, a phenomenon found to be mainly caused by two factors. First, the mannose-rich glycan of the IgG from Pichia, while slightly thermally destabilizing the IgG, strongly inhibited its aggregation susceptibility, compared to the complex mammalian glycan. Second, on the Pichia-produced IgGs, amino acids belonging to the α-factor pre-pro sequence were left at the N-termini of both chains. These additional residues proved to considerably increase the temperature of the onset of aggregation and reduced the aggregate formation after extended incubation at elevated temperatures. The attachment of these residues to IgGs produced in cell culture confirmed their beneficial effect on the aggregation resistance. Secretion of IgGs with native N-termini in the yeast system became possible after systematic engineering of the precursor proteins and the processing site. Taken together, the present results will be useful for the successful production of full-length IgGs in Pichia, give indications on how to engineer aggregation-resistant IgGs and shed new light on potential biophysical effects of tag sequences in general.


Alignment-free phylogeny of whole genomes using underlying subwords.
With the progress of modern sequencing technologies a large number of complete genomes are now available. Traditionally the comparison of two related genomes is carried out by sequence alignment. There are cases where these techniques cannot be applied, for example if two genomes do not share the same set of genes, or if they are not alignable to each other due to low sequence similarity, rearrangements and inversions, or more specifically to their lengths when the organisms belong to different species. For these cases the comparison of complete genomes can be carried out only with ad hoc methods that are usually called alignment-free methods. In this paper we propose a distance function based on subword compositions called Underlying Approach (UA). We prove that the matching statistics, a popular concept in the field of string algorithms able to capture the statistics of common words between two sequences, can be derived from a small set of "independent" subwords, namely the irredundant common subwords. We define a distance-like measure based on these subwords, such that each region of genomes contributes only once, thus avoiding to count shared subwords a multiple number of times. In a nutshell, this filter discards subwords occurring in regions covered by other more significant subwords. The Underlying Approach (UA) builds a scoring function based on this set of patterns, called underlying. We prove that this set is by construction linear in the size of input, without overlaps, and can be efficiently constructed. Results show the validity of our method in the reconstruction of phylogenetic trees, where the Underlying Approach outperforms the current state of the art methods. Moreover, we show that the accuracy of UA is achieved with a very small number of subwords, which in some cases carry meaningful biological information. http://www.dei.unipd.it/∼ciompin/main/underlying.html.


XB130, regulated by miR-203, miR-219, and miR-4782-3p, mediates the proliferation and metastasis of non-small-cell lung cancer cells.
XB130 is a novel adapter protein that behaves as a tumor promoter or suppressor mediating cell proliferation and metastasis in the development of different human tumors. Altered expression of XB130 has been verified in human non-small cell-lung cancer (NSCLC). However, the exact effect of XB130 on NSCLC is not well-understood. In this study, we investigated the biological function and posttranscriptional regulation of XB130 in NSCLC. First, the effects of XB130 silence on NSCLC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) were examined. Then the targeting relationship between XB130 and miR-203, miR-219, or miR-4782-3p was demonstrated by dual-luciferase reporter assay. Finally, the effects of miR-203, miR-219, and miR-4782-3p on NSCLC cell function were studied, respectively. We found that XB130 silence significantly inhibited cell growth, migration and invasion, and reversed EMT. Furthermore, XB130 was posttranscriptionally regulated by miR-203, miR-219, and miR-4782-3p. Overexpression of miR-203, miR-219, or miR-4782-3p inhibited cell growth, migration and invasion, and reversed EMT, just like the role of XB130 in NSCLC cells, whereas the suppressive effects of microRNA (miRNA) overexpression were weakened by miRNA inhibitors or ectopic expression of XB130 in NSCLC cells. These data demonstrate that XB130 is posttranscriptionally regulated by miR-203, miR-219, and miR-4782-3p and mediates the proliferation and metastasis of NSCLC cells.


RBM24 promotes U1 snRNP recognition of the mutated 5' splice site in the IKBKAP gene of familial dysautonomia.
The 5' splice site mutation (IVS20+6T>C) of the inhibitor of κ light polypeptide gene enhancer in B cells, kinase complex-associated protein (IKBKAP) gene in familial dysautonomia (FD) is at the sixth intronic nucleotide of the 5' splice site. It is known to weaken U1 snRNP recognition and result in an aberrantly spliced mRNA product in neuronal tissue, but normally spliced mRNA in other tissues. Aberrantly spliced IKBKAP mRNA abrogates IKK complex-associated protein (IKAP)/elongator protein 1 (ELP1) expression and results in a defect of neuronal cell development in FD. To elucidate the tissue-dependent regulatory mechanism, we screened an expression library of major RNA-binding proteins (RBPs) with our mammalian dual-color splicing reporter system and identified RBM24 as a regulator. RBM24 functioned as a cryptic intronic splicing enhancer binding to an element (IVS20+13-29) downstream from the intronic 5' splice site mutation in the IKBKAP gene and promoted U1 snRNP recognition only to the mutated 5' splice site (and not the wild-type 5' splice site). Our results show that tissue-specific expression of RBM24 can explain the neuron-specific aberrant splicing of IKBKAP exon 20 in familial dysautonomia, and that ectopic expression of RBM24 in neuronal tissue could be a novel therapeutic target of the disease.


Transverse mode discrimination in long-wavelength wafer-fused vertical-cavity surface-emitting lasers by intra-cavity patterning.
Transverse mode discrimination is demonstrated in long-wavelength wafer-fused vertical-cavity surface-emitting lasers using ring-shaped air gap patterns at the fused interface between the cavity and the top distributed Bragg reflector. A significant number of devices with varying pattern dimensions was investigated by on-wafer mapping, allowing in particular the identification of a design that reproducibly increases the maximal single-mode emitted power by about 30 %. Numerical simulations support these observations and allow specifying optimized ring dimensions for which higher-order transverse modes are localized out of the optical aperture. These simulations predict further enhancement of the single-mode properties of the devices with negligible penalty on threshold current and emitted power.


The reactivities and ionization properties of the active-site dithiol groups of mammalian protein disulphide-isomerase.
1. The number of reactive thiol groups in mammalian liver protein disulphide-isomerase (PDI) in various conditions was investigated by alkylation with iodo[14C]acetate. 2. Both the native enzyme, as isolated, and the urea-denatured enzyme contained negligible reactive thiol groups; the enzyme reduced with dithiothreitol contained two groups reactive towards iodoacetic acid at pH 7.5, and up to five reactive groups were detectable in the reduced denatured enzyme. 3. Modification of the two reactive groups in the reduced native enzyme led to complete inactivation, and the relationship between the loss of activity and the extent of modification was approximately linear. 4. Inactivation of PDI by alkylation of the reduced enzyme followed pseudo-first-order kinetics; a plot of the pH-dependence of the second-order rate constant for inactivation indicated that the essential reactive groups had a pK of 6.7 and a limiting second-order rate constant at high pH of 11 M-1.s-1. 5. Since sequence data on PDI show the presence within the polypeptide of two regions closely similar to thioredoxin, the data strongly indicate that these regions are chemically and functionally equivalent to thioredoxin. 6. The activity of PDI in thiol/disulphide interchange derives from the presence of vicinal dithiol groups in which one thiol group of each pair has an unusually low pK and high nucleophilic reactivity at physiological pH.


A comprehensive strategy to discover inhibitors of the translesion synthesis DNA polymerase κ.
Human DNA polymerase kappa (pol κ) is a translesion synthesis (TLS) polymerase that catalyzes TLS past various minor groove lesions including N(2)-dG linked acrolein- and polycyclic aromatic hydrocarbon-derived adducts, as well as N(2)-dG DNA-DNA interstrand cross-links introduced by the chemotherapeutic agent mitomycin C. It also processes ultraviolet light-induced DNA lesions. Since pol κ TLS activity can reduce the cellular toxicity of chemotherapeutic agents and since gliomas overexpress pol κ, small molecule library screens targeting pol κ were conducted to initiate the first step in the development of new adjunct cancer therapeutics. A high-throughput, fluorescence-based DNA strand displacement assay was utilized to screen ∼16,000 bioactive compounds, and the 60 top hits were validated by primer extension assays using non-damaged DNAs. Candesartan cilexetil, manoalide, and MK-886 were selected as proof-of-principle compounds and further characterized for their specificity toward pol κ by primer extension assays using DNAs containing a site-specific acrolein-derived, ring-opened reduced form of γ-HOPdG. Furthermore, candesartan cilexetil could enhance ultraviolet light-induced cytotoxicity in xeroderma pigmentosum variant cells, suggesting its inhibitory effect against intracellular pol κ. In summary, this investigation represents the first high-throughput screening designed to identify inhibitors of pol κ, with the characterization of biochemical and biologically relevant endpoints as a consequence of pol κ inhibition. These approaches lay the foundation for the future discovery of compounds that can be applied to combination chemotherapy.


Perturbation of cationic equilibrium by cucurbit-7-uril.
The effect of cucurbit-7-uril (CB-7) on a cationic mixture with same charge has been investigated by studying monocationic mixtures of 2-(4'-N,N-dimethylaminophenyl)imidazo[4,5-b]pyridine (DMAPIP-b) and 2-(4'-N,N-dimethylaminophenyl)imidazo[4,5-c]pyridine (DMAPIP-c). The pKa of both the guests increases in CB-7. DMAPIP-b forms all three monocations in the ground and the excited states in aqueous as well as in CB-7 solution. However, CB-7 shifts the equilibrium more towards the less polar MC2 and MC3. On the other hand, DMAPIP-c exists only as MC1 and MC3 in aqueous solution, however, in CB-7 it exists not only as MC1 and MC3 but also as MC2 in CB-7 in the ground state. In the excited state, as in aqueous solution MC1 forms MC2 by biprotonic phototautomerism in CB-7. The association constants of monocations suggest a pyridyl nitrogen position dependence. All the MC-CB-7 complexes are optimized by density functional theory (DFT).


Dietary intake and supplement use of vitamins C and E and upper respiratory tract infection.
Antioxidants are regulators of immune function and may play a role in upper respiratory tract infections (URTI). We investigated the potential effects of dietary intake from food and supplement use of vitamins C and E on the risk of self-reported URTI. We conducted a population-based cohort study of 1509 Swedish men and women ages 20 to 60 with a follow-up period of 4 months. Participants reported a total of 1181 occurrences of URTI. Poisson regression model was used to control for age, sex, and other confounding factors. Among women, we found that the incidence rate ratio (IRR) for high intake of vitamin C (>200 mg/d) from food was 0.69 (95% CI 0.49-0.98) compared with low intake (<100 mg/d). This association was not seen among men, for whom the IRR was 1.16 (95% CI 0.79-1.70) for high intake of vitamin C (>150 mg/d) compared with low intake (<50 mg/d). We saw no protective effect of vitamin E from food among either men or women, but a possible protective effect of vitamin C and E supplement use among men (vitamin C, 0.69 [95% CI 0.47-1.02]; vitamin E, 0.56 [95% CI 0.33-0.95]), although not among women. The present study is the first observational study to suggest that intake of vitamin C from food is sufficient to lower the risk of URTI among women. In addition, it appears that supplement use of vitamin E and vitamin C may reduce the risk of URTI among men, who overall had a lower intake of vitamin C from food than women.


Lipid Supplements and Clinical Aspects of Tear Film in Habitual Lens Wearers.
To establish the effect of lipid supplements on the tear lipid layer and their influence on lens wear comfort in habitual lens wearers. Forty habitual soft contact lens wearers were recruited to a double-masked, randomized crossover trial. An emulsion drop containing phosphatidylglycerine (Systane Balance; Alcon) and a saline drop as a placebo or a liposomal spray containing phosphatidylcholine (Tears again; BioRevive) and a saline spray as a placebo were used three times a day for 2 weeks with 48 hours washout between each intervention. Ocular comfort, lipid layer grade, and stability of the tear film using a Tearscope and tear evaporation rate using a modified VapoMeter were assessed after 6 hours of lens wear with lenses in situ. Neither of the lipid supplements improved lens wear comfort compared to baseline. The noninvasive surface drying time significantly reduced with the placebo spray at day 1 (P = .002) and day 14 (P = .01) whereas the lipid spray had no effect. With the lipid drop and placebo, noninvasive surface drying time was unchanged compared to baseline (P > .05) on day 1, but by day 14, noninvasive surface drying time was reduced with the lipid drop (P = .02) and placebo (P < .001). Symptomatic wearers showed shorter noninvasive surface drying time compared to asymptomatic wearers with the spray treatment on both days (P = .03) but not with the lipid drop (P = .64). The placebo drop significantly changed the lipid layer distribution (P = .03) with a higher percentage of thinner patterns compared to the baseline distribution at day 14. A weak but significant correlation was shown between ocular comfort and noninvasive surface drying time (r = -0.21, P = .003) and tear evaporation rate (r = 0.19, P = .008). Ocular comfort was not associated with lipid layer patterns (r = 0.13, P = .06). Ocular comfort during contact lens wear improved with increased tear film stability and a reduced tear evaporation rate. However, the lipid supplements did not improve ocular comfort from baseline.


Donor-Site Morbidity and Functional Status following Medial Femoral Condyle Flap Harvest.
The medial femoral condyle free vascularized bone flap is a valuable alternative to other types of vascularized bone grafts. The donor-site morbidity and functional outcomes after flap harvest have not been fully appreciated. The authors report the postoperative outcomes and analyze the impact of increasing the size of the flap on knee donor-site morbidity. A retrospective chart review of patients who underwent medial femoral condyle flap surgery between 2001 and 2012 at their institution was performed. The size of the flap was stratified, based on the largest dimension, into three groups. Demographics, outcomes, and complications related to the flap donor site were recorded and analyzed. Subsequently, functional status was assessed by administering a validated condition-specific measure. A univariate logistic regression analysis was performed, and results were analyzed. A total of 75 patients were identified. Average age was 29.5 ± 15.2 years. Average follow-up time was 13 months. The overall complication rate was 18.6 percent. Paresthesia in the saphenous nerve distribution was the most common complication. Increasing the size of the flap resulted in a significant elevation in complication risk (p < 0.05). A total of 47 patients completed the Lower Extremity Functional Scale questionnaire. The average Lower Extremity Functional Scale score was 72.12 ± 14.18. Fifty-one percent (n = 24) scored 80 points, indicating a normal level of function on average. The medial femoral condyle flap has overall acceptable donor-site morbidity, with a good level of function postoperatively. Larger flaps are associated with a greater number of complications. Therapeutic, III.


A new method of urethroplasty for prevention of fistula in female-to-male gender reassignment surgery.
One of the primary goals of the phalloplasty for female-to-male transsexuals is to gain the voiding ability in the standing position. However, achieving the competence of urethra, sensation and rigidity of the neophallus is still a significant challenge. Serious complications such as urethral fistula, obstruction and stricture were encountered in this surgery. In experienced hands, this seems to be associated with urethroplasty technique. The authors performed phalloplasty with radial forearm osteocutaneous free flap method in 70 patients of female-to-male transsexuals. In 38 cases which were enrolled before 2001, we had carried out the urethroplasty by our own method, but since 2001, we have applied the modified method of urethroplasty to reduce the incidence of urethrocutaneous fistula. Thirty-four cases have undergone a new modified method of ours. For construction of the urethra, an anteriorly based vaginal wall flap and labium minoral flaps were used in this technique. In our new method series, 1 case (1.4%) of flap loss occurred after phalloplasty. The incidence of urethrocutaneous fistula was 30%. Before the year 2001, of 38 patients, there were 14 cases (36.8%) who developed urethrocutaneous fistula. On the other hand, 7 of 32 patients (21.9%) who underwent urethroplasty by the modified labium minoral flap and anteriorly based vaginal flap had urethrocutaneous fistula after 2001. One-stage total phalloplasty and urethroplasty is associated with a significant increase of urethral fistula and obstruction. However, the urethrocutaneous fistula at the level of the female external urethral orifice can be successfully reduced using this new method.


HER2 Regulates Cancer Stem Cell Activities via the Wnt Signaling Pathway in Gastric Cancer Cells.
Human epidermal growth factor 2 (HER2) gene overexpression in breast carcinoma cell lines has been shown to drive mammary carcinogenesis and tumor growth and invasion through its effects on mammary stem cells. Therefore, we investigated the mechanism by which HER2 regulates cancer stem cell (CSC) activity in gastric cancer cells. HER2 was transfected into MKN28 gastric cancer cells, and its role in regulating CSC activity was determined by characterizing the HER2-overexpressing cells. The sphere formation assay revealed that the sphere sizes and frequency of sphere formation were significantly greater for the HER2-overexpressing cells than for the MKN28 control cells. The CSC markers Oct-4 and BMI1 were more highly expressed in the HER2-overexpressing cells, as were the EMT markers. This was accompanied by a significant enhancement in cellular invasion of the Matrigel and migration. The E-cadherin level was significantly downregulated, and the mesenchymal marker Snail upregulated, in the HER2-transfected cells. HER2 overexpression activated the well-characterized CSC-associated Wnt/β-catenin signaling pathway, as shown by the luciferase assay. After treatment of these cells with the Wnt signal inhibitor PRI-724, the BMI1 and Oct-4 levels were decreased for 24 h and Snail was also downregulated. Immunofluorescence staining revealed the significant restoration of E-cadherin levels in the HER2-transfected cells after PRI-724 treatment. These results established a role for HER2 in regulating gastric CSC activity, with Wnt/β-catenin signaling being mediated via a HER2-dependent pathway. In summary, HER2-overexpressing gastric cancer cells exhibited increased stemness and invasiveness and were regulated by Wnt/β-catenin signaling.


Long-term CD4+ memory T cells from the spleen lack MEL-14, the lymph node homing receptor.
We have characterized the surface phenotype and function of long-lived, Ag-specific memory CD4+ T cells generated in vivo by immunization with keyhole limpet hemocyanin (KLH). CD4+ T cells from the spleens of mice primed more than 2 mo previously with KLH, produced high levels of IL-2 and IL-3, and low levels of IL-4 and IFN-gamma in response to in vitro restimulation with specific Ag. The KLH-primed T cells mediated carrier-specific helper activity for the antibody production by NIP-primed B cells in secondary in vitro responses to NIP-KLH. Subsets of CD4+ T cells from KLH-primed mice were isolated on the basis of surface CD45RB (23G2) by magnetic separation and were examined for functional capacity in several assays of Ag-specific recall. Virtually all of the secretion of IL-2, IL-3, IL-4, and IFN-gamma in response to restimulation with Ag in vitro was associated with, and considerably enriched in, the CD45RB- subset of CD4+ T cells. Similarly, carrier-specific helper function and Ag-specific proliferation in vitro were also confined to the CD45RB-, CD4+ subset of T cells, confirming the previous association of this surface phenotype with memory Th cell activity. We also examined expression of the lymphocyte homing receptor, MEL-14 (gp90MEL), which is required for lymphocyte extravasation to peripheral lymph nodes and is present in high levels on naive T cells. MEL-14 positive and negative subsets of CD4+ T cells from long term KLH-primed mice were evaluated for Ag-specific memory function in terms of lymphokine production, Ag-induced proliferation, and helper activity. Each of these functions was associated exclusively with the MEL-14- subset of CD4+ T cells, which exhibited responses comparable to the CD45RB- subset. These data indicate that memory Th cell function in the spleen is contained within the MEL-14-, CD45RB- subset of CD4+ T cells and suggest that memory helper cells may have different patterns of recirculation from naive T cells.


The impact of case diagnosis coverage and diagnosis delays on the effectiveness of antiviral strategies in mitigating pandemic influenza A/H1N1 2009.
Neuraminidase inhibitors were used to reduce the transmission of pandemic influenza A/H1N1 2009 at the early stages of the 2009/2010 pandemic. Policies for diagnosis of influenza for the purposes of antiviral intervention differed markedly between and within countries, leading to differences in the timing and scale of antiviral usage. The impact of the percentage of symptomatic infected individuals who were diagnosed, and of delays to diagnosis, for three antiviral intervention strategies (each with and without school closure) were determined using a simulation model of an Australian community. Epidemic characteristics were based on actual data from the A/H1N1 2009 pandemic including reproduction number, serial interval and age-specific infection rate profile. In the absence of intervention an illness attack rate (AR) of 24.5% was determined from an estimated R(0) of 1.5; this was reduced to 21%, 16.5% or 13% by treatment-only, treatment plus household prophylaxis, or treatment plus household plus extended prophylaxis antiviral interventions respectively, assuming that diagnosis occurred 24 hours after symptoms arose and that 50% of symptomatic cases were diagnosed. If diagnosis occurred without delay, ARs decreased to 17%, 12.2% or 8.8% respectively. If 90% of symptomatic cases were diagnosed (with a 24 hour delay), ARs decreased to 17.8%, 11.1% and 7.6%, respectively. The ability to rapidly diagnose symptomatic cases and to diagnose a high proportion of cases was shown to improve the effectiveness of all three antiviral strategies. For epidemics with R(0)< = 1.5 our results suggest that when the case diagnosis coverage exceeds ∼70% the size of the antiviral stockpile required to implement the extended prophylactic strategy decreases. The addition of at least four weeks of school closure was found to further reduce cumulative and peak attack rates and the size of the required antiviral stockpile.


[Effects of acupuncture at "Zusanli" (ST 36) on eosinophil apoptosis and related gene expression in rats with asthma].
To observed efficacy differences of acupuncture at "Zusanli" (ST 36) in rats with asthma and asthma with spleen-deficency, so as to investigate the therapeutic mechanism. Sixty SD rats were randomly divided into 5 groups according to their weight, named as an asthma with spleen-deficency group (group A), an acupuncture on asthma with spleen-deficency group (group B), an asthma group (group C), an acupuncture on asthma group (group D) and a control group. The rat models with spleen-deficiency in the first two groups were set up by TCM, then the rats of asthma model in the first four groups were induced by egg albumin, but the control group was treated by the same dose of saline. The group B and the group D were both treated with acupuncture at "Zusanli" (ST 36), once each day for 8 days, and the other groups remained unhandled. The mRNA expressions of Fas and Bcl-2 in the lung tissues were detected by hybridization in situ and apoptosis was detected by TUNEL (terminal dexynucleotidyl transferase-mediated dutp nick end labeling). Compared with the control group, in both the group A and the group C, the expression of Fas mRNA significantly decreased, but the expression of Bcl-2 mRNA significantly increased (all P < 0.01), and eosinophils (EOS) counts significantly increased, but EOS apoptosis rate significantly decreased (all P < 0.01). Compared with the group C, in the group A, the expressions of Fas mRNA significantly decreased, but the expressions of Bcl-2 mRNA and EOS counts significantly increased (all P < 0.01). At the same time, compared with the corresponding asthma groups, in both acupuncture groups, Fas mRNA expression obviously increased, Bcl-2 mRNA expression was significantly reduced (all P < 0.01), EOS counts remarkably decreased and EOS apoptosis rate significantly increased (all P < 0.01). There were no significant differences in the expressions of Fas mRNA and Bcl-2 mRNA between the two acupuncture groups (both P > 0.05), but compared with group B,in the group D, EOS counts significantly decreased and EOS apoptosis rate significantly increased (both P < 0.01). Acupuncture at "Zusanli" (ST 36) can regulate the disorders of Fas mRNA and Bcl-2 mRNA expression in the state of both asthma and asthma with spleen-deficency, promote EOS apoptosis so as to inhibit the development of inflammatory reaction of asthma, showing that acupuncture at "Zusanli" (ST 36) has certain advantages on regulation of related gene of EOS in asthma with spleen-deficency.


Cold-adapted arsenite oxidase from a psychrotolerant Polaromonas species.
Polaromonas sp. str. GM1 is an aerobic, psychrotolerant, heterotrophic member of the Betaproteobacteria and is the only isolate capable of oxidising arsenite at temperatures below 10 °C. Sequencing of the aio gene cluster in GM1 revealed the presence of the aioB and aioA genes, which encode the arsenite oxidase but the regulatory genes typically found upstream of aioB in other members of the Proteobacteria were absent. The GM1 Aio was purified to homogeneity and was found to be a heterodimer. The enzyme contained Mo and Fe as cofactors and had, using the artificial electron acceptor 2,6-dichlorophenolindophenol, a Km for arsenite of 111.70 ± 0.88 μM and a Vmax of 12.16 ± 0.30 U mg(-1), which is the highest reported specific activity for any known Aio. The temperature-activity profiles of the arsenite oxidases from GM1 and the mesophilic betaproteobacterium Alcaligenes faecalis were compared and showed that the GM1 Aio was more active at low temperatures than that of A. faecalis. A homology model of the GM1 Aio was made using the X-ray crystal structure of the Aio from A. faecalis as the template. Structural changes that account for cold adaptation were identified and it was found that these resulted in increased enzyme flexibility and a reduction in the hydrophobicity of the core.


Metformin improves cancer immunotherapy by directly rescuing tumor-infiltrating CD8 T lymphocytes from hypoxia-induced immunosuppression.
Despite their revolutionary success in cancer treatment over the last decades, immunotherapies encounter limitations in certain tumor types and patients. The efficacy of immunotherapies depends on tumor antigen-specific CD8 T-cell viability and functionality within the immunosuppressive tumor microenvironment, where oxygen levels are often low. Hypoxia can reduce CD8 T-cell fitness in several ways and CD8 T cells are mostly excluded from hypoxic tumor regions. Given the challenges to achieve durable reduction of hypoxia in the clinic, ameliorating CD8 T-cell survival and effector function in hypoxic condition could improve tumor response to immunotherapies. Activated CD8 T cells were exposed to hypoxia and metformin and analyzed by fluorescence-activated cell sorting for cell proliferation, apoptosis and phenotype. In vivo, metformin was administered to mice bearing hypoxic tumors and receiving either adoptive cell therapy with tumor-specific CD8 T cells, or immune checkpoint inhibitors; tumor growth was followed over time and CD8 T-cell infiltration, survival and localization in normoxic or hypoxic tumor regions were assessed by flow cytometry and immunofluorescence. Tumor oxygenation and hypoxia were measured by electron paramagnetic resonance and pimonidazole staining, respectively. We found that the antidiabetic drug metformin directly improved CD8 T-cell fitness in hypoxia, both in vitro and in vivo. Metformin rescued murine and human CD8 T cells from hypoxia-induced apoptosis and increased their proliferation and cytokine production, while blunting the upregulation of programmed cell death protein 1 and lymphocyte-activation gene 3. This appeared to result from a reduced production of reactive oxygen species, due to the inhibition of mitochondrial complex I. Differently from what others reported, metformin did not reduce tumor hypoxia, but rather increased CD8 T-cell infiltration and survival in hypoxic tumor areas, and synergized with cyclophosphamide to enhance tumor response to adoptive cell therapy or immune checkpoint blockade in different tumor models. This study describes a novel mechanism of action of metformin and presents a promising strategy to achieve immune rejection in hypoxic and immunosuppressive tumors, which would otherwise be resistant to immunotherapy.


Effect of magnesium on the osteogenesis of normal human osteoblasts.
Biomaterials containing magnesium are used for implants and bone regeneration. However, mechanisms underlying the biologic effects of magnesium are still largely unknown and have not been examined on normal human osteoblasts. This study was designed to test the effect of supplemented Mg2+ concentrations between 0.5 mM and 16 mM on the osteogenic behaviors of normal human primary osteoblasts. Human primary osteoblasts were cultured in the groups with various concentrations of supplemented magnesium for various time intervals. Cell proliferation was measured using crystal violet staining. Degree of Alkaline Phosphatase (ALP) activity was measured by fluorometric assay. Expression of osteocalcin was measured by immunosorbent assay. Mineralization of cultures was determined by Alizarin Red S staining. Results showed that initial cell attachment efficiency was not affected by supplemented Mg2+ (P > 0.05). At 21 days, proliferation rates increased in groups containing 0.5 mM-4 mM supplemented Mg2+ and decreased in groups of supplemented 8 mM and 16 mM Mg2+. ALP activity and osteocalcin expression were upregulated in groups of supplemented Mg2+ between 0.5 mM-2.0 mM (P < 0.05), but downregulated in groups with supplemented Mg2+ concentrations of 4mM and above (P < 0.05). Cultures with 1 mM and 2 mM supplemented Mg2+ showed upregulated mineralization activity compared to the control (P < 0.05), but downregulated in groups with supplemented Mg2+ concentrations of 4 mM and above (P < 0.05). The present study based on an experimental design demonstrated the impact of 2 mM supplemented Mg2+ on induced-proliferation and differentiation of normal human osteoblasts.


The efficacy of topical Royal Jelly on diabetic foot ulcers healing: A case series.
Foot ulcers and infections are the major sources of morbidity in individuals with diabetes mellitus. This study aimed to evaluate the efficacy of topical Royal Jelly (a worker honey bee product) on healing diabetic foot ulcers. Diabetic patients with foot ulcers that were referred to our clinic at Khorshid Hospital, Isfahan, Iran; were evaluated three times a week and treated according to standard treatments consisted of offloading, infection control, vascular improvement and debridement if required. In addition, all ulcers were measured and then topical sterile 5% Royal Jelly was applied on the total surface area of the wounds. Eventually, they were covered with sterile dressings. Each patient was followed for a period of three months or until the complete healing. A total of eight patients were enrolled in this study. Of these, two had two ulcers and, therefore, ten ulcers were evaluated. Two ulcers were excluded. Seven of the remained eight ulcers healed. Mean duration of complete healing was 41 days. One ulcer did not completely heal but improved to 40% smaller in length, 32% in width and 28% in depth. The mean length, width and depth reduction rates were 0.35 mm/day, 0.28 mm/day and 0.11 mm/day, respectively. Royal Jelly dressing may be an effective method for treating diabetic foot ulcers besides standard treatments.


Do compression garments enhance the active recovery process after high-intensity running?
Lovell, DI, Mason, DG, Delphinus, EM, and McLellan, CP. Do compression garments enhance the active recovery process after high-intensity running? J Strength Cond Res 25(12): 3264-3268, 2011-This study examined the effect of wearing waist-to-ankle compression garments (CGs) on active recovery after moderate- and high-intensity submaximal treadmill running. Twenty-five male semiprofessional rugby league players performed two 30-minute treadmill runs comprising of six 5-minute stages at 6 km·h, 10 km·h, approximately 85% VO(2)max, 6 km·h as a recovery stage followed by approximately 85% VO(2)max and 6 km·h wearing either CGs or regular running shorts in a randomized counterbalanced order with each person acting as his own control. All stages were followed by 30 seconds of rest during which a blood sample was collected to determine blood pH and blood lactate concentration [La]. Expired gases and heart rate (HR) were measured during the submaximal treadmill tests to determine metabolic variables with the average of the last 2 minutes used for data analysis. The HR and [La] were lower (p ≤ 0.05) after the first and second 6 km·h recovery bouts when wearing CGs compared with when wearing running shorts. The respiratory exchange ratio (RER) was higher and [La] lower (p ≤ 0.05) after the 10 km·h stage, and only RER was higher after both 85% VO(2)max stages when wearing CGs compared with when wearing running shorts. There was no difference in blood pH at any exercise stage when wearing the CGs and running shorts. The results of this study indicate that the wearing of CGs may augment the active recovery process in reducing [La] and HR after high-intensity exercise but not effect blood pH. The ability to reduce [La] and HR has important consequences for many sports that are intermittent in nature and consist of repeated bouts of high-intensity exercise interspersed with periods of low-intensity exercise or recovery.


Direct visualization of light confinement and standing wave in THz Fabry-Perot resonator with Bragg mirrors.
We report for the first time the ability to perform time resolved imaging of terahertz (THz) waves propagating within a Fabry-Perot resonator on a LiNbO3 slab. Electro-optic effect is used to record the full spatiotemporal evolution of THz fields inside the resonator. In addition to revealing the real-space behavior, the data further demonstrate the confinement and the standing wave modes of THz in the cavity in frequency domain. The experimental results are in good agreement with numerical simulations. Using the coherent imaging technique to gain real-time information about a resonator system provides a unique path to study the physics of optical cavity.


La pulvérisation par ultra-sons appliquée à la spectrometrie d'absorption atomique.
Ultrasonic nebulization applied to atomic absorption spectrometry gives a sensitivity ten to twenty times better than that which can be obtained by classical pneumatic nebulization. This gain in sensitivity is primarily due to the quantity of aerosol introduced into the flame, but is equally due to the fineness of the droplets that are formed. The study of certain parameters (frequency, power applied to the emitter, geometry of the nebulizing chamber, gas flow) has led to the specification of an ultrasonic nebulizer that can be conveniently used for routine analyses.


Mental imagery for full and upper human bodies: common right hemisphere activations and distinct extrastriate activations.
The processing of human bodies is important in social life and for the recognition of another person's actions, moods, and intentions. Recent neuroimaging studies on mental imagery of human body parts suggest that the left hemisphere is dominant in body processing. However, studies on mental imagery of full human bodies reported stronger right hemisphere or bilateral activations. Here, we measured functional magnetic resonance imaging during mental imagery of bilateral partial (upper) and full bodies. Results show that, independently of whether a full or upper body is processed, the right hemisphere (temporo-parietal cortex, anterior parietal cortex, premotor cortex, bilateral superior parietal cortex) is mainly involved in mental imagery of full or partial human bodies. However, distinct activations were found in extrastriate cortex for partial bodies (right fusiform face area) and full bodies (left extrastriate body area). We propose that a common brain network, mainly on the right side, is involved in the mental imagery of human bodies, while two distinct brain areas in extrastriate cortex code for mental imagery of full and upper bodies.


Is Parent-Adolescent Drug Talk Always Protective? Testing A New Scale of Drug Talk Styles in Relation to Adolescent Personal Norms, Parental Injunctive Norms, Substance Use Intentions, and Behaviors.
According to parent-offspring drug talk (PODT) model, the specificity of drug talk styles is identified by the timing and directness of communication about substance use between parent and adolescent (e.g., situated direct, ongoing direct, situated indirect, and ongoing indirect talk style). Given the limitation of the original, single item measure for drug talk styles with a categorical response option, the current study proposed a new scale of drug talk styles and tested its concurrent validity. Using cross-sectional survey data (N = 2,035), confirmatory factor analysis was conducted to test the factor structure of the scale. Consistent with PODT model, the analysis yielded support for a four-factor structure for the drug talk styles. Next, a path analysis was employed to validate the scale. The findings suggested that situated direct talk was positively related to personal anti-substance-use norms and parental anti-substance-use injunctive norms but negatively related to smoking intention, as well as alcohol use in the past 30 days. Ongoing direct talk, however, was found to be positively associated with alcohol and marijuana use in the past 30 days. Research implications and future directions are discussed.


Pemphigus vulgaris in Jewish patients is associated with HLA-A region genes: mapping by microsatellite markers.
Pemphigus vulgaris (PV) is the most severe autoimmune blistering disorder of the skin that is mediated by circulating autoantibodies against desmoglein 3 (Dsg3). It has been reported that in Jews the associated haplotype in PV is human leukocyte antigen (HLA) B38, DRB1*0402, DQB1*0302. Significant associations with HLA were observed also in non-Jews. Dsg3-specific T-cell responses were detected in PV patients but also in healthy individuals who were either carriers of the PV-associated DRB1*0402 allele or alleles that share similar or identical peptide binding motifs to DRB1*0402. This suggests that genes other than the classical major histocompatibility complex (MHC) genes are associated with the development of the autoimmune response. We used 16 microsatellite probes that span the entire MHC region to screen DNA samples from 38 PV patients and 76 healthy controls. Results demonstrated that some markers were associated with class II region including a TAP associated marker. However, four probes, D6S265, C_527, D6S510, and MOGC, which are all mapped to the region of HLA-A, were highly associated with PV. These results suggest that a gene, or genes in the class I region are important in the initiation of the autoimmune cascade. Activation/suppression of these genes might act as the trigger mechanism that starts the autoimmune destructive process.


The distribution of SNPs in human gene regulatory regions.
As a result of high-throughput genotyping methods, millions of human genetic variants have been reported in recent years. To efficiently identify those with significant biological functions, a practical strategy is to concentrate on variants located in important sequence regions such as gene regulatory regions. Analysis of the most common type of variant, single nucleotide polymorphisms (SNPs), shows that in gene promoter regions more SNPs occur in close proximity to transcriptional start sites than in regions further upstream, and a disproportionate number of those SNPs represent nucleotide transversions. Additionally, the number of SNPs found in the predicted transcription factor binding sites is higher than in non-binding site sequences. Current information about transcription factor binding site sequence patterns may not be exhaustive, and SNPs may be actively involved in influencing gene expression by affecting the transcription factor binding sites.


Ultrasound Imaging for Muscle Variations: Digastric Flexor Carpi Ulnaris, Gastrocnemius Tertius, and Supernumerary Fibularis Longus in an Asymptomatic Family.
Anatomical variations of the muscles are seen with different frequencies. Although most of them are asymptomatic; in certain cases, their existence requires attention with regard to entrapment syndromes, mass lesions, botulinum toxin injections, and tendon transfers. Herein, as ultrasound imaging is a convenient method for muscle imaging, it can easily be used in daily practice for prompt understanding of such muscular variations. In this report, we demonstrated and discussed a similar scenario in an asymptomatic individual (and his close family members) using ultrasound imaging for scanning the digastric flexor carpi ulnaris, gastrocnemius tertius, and supernumerary fibularis longus muscles.


Lingual Pressure as a Clinical Indicator of Swallowing Function in Parkinson's Disease.
Swallowing impairment, or dysphagia, is a known contributor to reduced quality of life, pneumonia, and mortality in Parkinson's disease (PD). However, the contribution of tongue dysfunction, specifically inadequate pressure generation, to dysphagia in PD remains unclear. Our purpose was to determine whether lingual pressures in PD are (a) reduced, (b) reflect medication state, or are (c) consistent with self-reported diet and swallowing function. Twenty-eight persons with idiopathic PD (PwPD) and 28 age- and sex-matched controls completed lingual pressure tasks with the Iowa Oral Performance Instrument. PwPD were tested during practically defined ON and OFF dopaminergic medication states. Participants were also stratified into three sex- and age-matched cohorts (7 men, 5 women): (a) controls, (b) PwPD without self-reported dysphagia symptoms or diet restrictions, and (c) PwPD with self-reported dysphagia symptoms with or without diet restrictions. PwPD exhibited reduced tongue strength and used elevated proportions of tongue strength during swallowing compared with controls (p < .05) without an effect of medication state (p > .05). Reduced tongue strength distinguished PwPD with self-reported dysphagia symptoms from PwPD without reported symptoms or diet restrictions (p = .045) and controls (p = .002). Tongue strength was significantly reduced in PwPD and did not differ by medication state. Tongue strength differentiated between PwPD with and without self-reported swallowing symptoms. Therefore, measures of tongue strength and swallowing pressures may serve as clinical indicators for further dysphagia evaluation and may promote early diagnosis and management of dysphagia in PD.


Conserved motifs involved in ATP hydrolysis by MalT, a signal transduction ATPase with numerous domains from Escherichia coli.
The signal transduction ATPases with numerous domains (STAND) are sophisticated signaling proteins that are related to AAA+ proteins and control various biological processes, including apoptosis, gene expression, and innate immunity. They function as tightly regulated switches, with the off and on positions corresponding to an ADP-bound, monomeric form and an ATP-bound, multimeric form, respectively. Protein activation is triggered by inducer binding to the sensor domain. ATP hydrolysis by the nucleotide-binding oligomerization domain (NOD) ensures the generation of the ADP-bound form. Here, we use MalT, an Escherichia coli transcription activator, as a model system to identify STAND conserved motifs involved in ATP hydrolysis besides the catalytic acidic residue. Alanine substitution of the conserved polar residue (H131) that is located two residues downstream from the catalytic residue (D129) blocks ATP hydrolysis and traps MalT in an active, ATP-bound, multimeric form. This polar residue is also conserved in AAA+. Based on AAA+ X-ray structures, we proposed that it is responsible for the proper positioning of the catalytic and the sensor I residues for the hydrolytic attack. Alanine substitution of the putative STAND sensor I (R160) abolished MalT activity. Substitutions of R171 impaired both ATP hydrolysis and multimerization, which is consistent with an arginine finger function and provides further evidence that ATP hydrolysis is primarily catalyzed by MalT multimers.


Proximate weather patterns and spring green-up phenology effect Eurasian beaver (Castor fiber) body mass and reproductive success: the implications of climate change and topography.
Low spring temperatures have been found to benefit mobile herbivores by reducing the rate of spring-flush, whereas high rainfall increases forage availability. Cold winters prove detrimental, by increasing herbivore thermoregulatory burdens. Here we examine the effects of temperature and rainfall variability on a temperate sedentary herbivore, the Eurasian beaver, Castor fiber, in terms of inter-annual variation in mean body weight and per territory offspring production. Data pertain to 198 individuals, over 11 years, using capture-mark-recapture. We use plant growth (tree cores) and fAPAR (a satellite-derived plant productivity index) to examine potential mechanisms through which weather conditions affect the availability and the seasonal phenology of beaver forage. Juvenile body weights were lighter after colder winters, whereas warmer spring temperatures were associated with lighter adult body weights, mediated by enhanced green-up phenology rates. Counter-intuitively, we observed a negative association between rainfall and body weight in juveniles and adults, and also with reproductive success. Alder, Alnus incana, (n = 68) growth rings (principal beaver food in the study area) exhibited a positive relationship with rainfall for trees growing at elevations >2 m above water level, but a negative relationship for trees growing <0.5 m. We deduce that temperature influences beavers at the landscape scale via effects on spring green-up phenology and winter thermoregulation. Rainfall influences beavers at finer spatial scales through topographical interactions with plant growth, where trees near water level, prone to water logging, producing poorer forage in wetter years. Unlike most other herbivores, beavers are an obligate aquatic species that utilize a restricted 'central-place' foraging range, limiting their ability to take advantage of better forage growth further from water during wetter years. With respect to anthropogenic climate change, interactions between weather variables, plant phenology and topography on forage growth are instructive, and consequently warrant examination when developing conservation management strategies for populations of medium to large herbivores.


An unusual location for cellular schwannoma.
Schwannomas are solitary benign tumours derived from peripheral nerve sheath that are difficult to diagnose using imaging only. Diagnoses are commonly confirmed with conventional surgical resection. Small proportion of cases are formed by cellular schwannoma usually seen at paravertebral, pelvic, retroperitoneal, or mediastinal location. This patient had a 3 x 2 cm hard mass at the dorsal side of proximal phalanx of the right third toe. Bone destruction was seen on radiography and MR imaging. Curettage after excision and grafting were performed. No complication was seen after surgery. The histopathology result was reported as cellular schwannoma. It was an atypical location for cellular schwannoma not previously described in literature.


A 3D interactive multi-object segmentation tool using local robust statistics driven active contours.
Extracting anatomical and functional significant structures renders one of the important tasks for both the theoretical study of the medical image analysis, and the clinical and practical community. In the past, much work has been dedicated only to the algorithmic development. Nevertheless, for clinical end users, a well designed algorithm with an interactive software is necessary for an algorithm to be utilized in their daily work. Furthermore, the software would better be open sourced in order to be used and validated by not only the authors but also the entire community. Therefore, the contribution of the present work is twofolds: first, we propose a new robust statistics based conformal metric and the conformal area driven multiple active contour framework, to simultaneously extract multiple targets from MR and CT medical imagery in 3D. Second, an open source graphically interactive 3D segmentation tool based on the aforementioned contour evolution is implemented and is publicly available for end users on multiple platforms. In using this software for the segmentation task, the process is initiated by the user drawn strokes (seeds) in the target region in the image. Then, the local robust statistics are used to describe the object features, and such features are learned adaptively from the seeds under a non-parametric estimation scheme. Subsequently, several active contours evolve simultaneously with their interactions being motivated by the principles of action and reaction-this not only guarantees mutual exclusiveness among the contours, but also no longer relies upon the assumption that the multiple objects fill the entire image domain, which was tacitly or explicitly assumed in many previous works. In doing so, the contours interact and converge to equilibrium at the desired positions of the desired multiple objects. Furthermore, with the aim of not only validating the algorithm and the software, but also demonstrating how the tool is to be used, we provide the reader reproducible experiments that demonstrate the capability of the proposed segmentation tool on several public available data sets.


Multilayer Architectures for Facial Action Unit Recognition.
In expression recognition and many other computer vision applications, the recognition performance is greatly improved by adding a layer of nonlinear texture filters between the raw input pixels and the classifier. The function of this layer is typically known as feature extraction. Popular filter types for this layer are Gabor energy filters (GEFs) and local binary patterns (LBPs). Recent work [1] suggests that adding a second layer of nonlinear filters on top of the first layer may be beneficial. However, it is unclear what is the best architecture of layers and selection of filters. In this paper, we present a thorough empirical analysis of the performance of single-layer and dual-layer texture-based approaches for action unit recognition. For the single hidden layer case, GEFs perform consistently better than LBPs, which may be due to their robustness to jitter and illumination noise as well as to their ability to encode texture at multiple resolutions. For dual-layer case, we confirm that, while small, the benefit of adding this second layer is reliable and consistent across data sets. Interestingly for this second layer, LBPs appear to perform better than GEFs.


Chitosan promotes cancer progression and stem cell properties in association with Wnt signaling in colon and hepatocellular carcinoma cells.
Cancer stem cells (CSCs), a small population of cancer cells, have been considered to be the origin of cancer initiation, recurrence, and metastasis. Tumor microenvironment provides crucial signals for CSCs to maintain stem cell properties and promotes tumorigenesis. Therefore, establishment of an appropriate cell culture system to mimic the microenvironment for CSC studies is an important issue. In this study, we grew colon and hepatocellular carcinoma (HCC) cells on chitosan membranes and evaluated the tumor progression and the CSC properties. Experimental results showed that culturing cancer cells on chitosan increased cell motility, drug resistance, quiescent population, self-renewal capacity, and the expression levels of stemness and CSC marker genes, such as OCT4, NANOG, CD133, CD44, and EpCAM. Furthermore, we demonstrated that chitosan might activate canonical Wnt/β-catenin-CD44 axis signaling in CD44positive colon cancer cells and noncanonical Wnt-STAT3 signaling in CD44negative HCC cells. In conclusion, chitosan as culture substrates activated the essential signaling of CSCs and promoted CSC properties. The chitosan culture system provides a convenient platform for the research of CSC biology and screening of anticancer drugs.


Thimerosal induces micronuclei in the cytochalasin B block micronucleus test with human lymphocytes.
Thimerosal is a widely used preservative in health care products, especially in vaccines. Due to possible adverse health effects, investigations on its metabolism and toxicity are urgently needed. An in vivo study on chronic toxicity of thimerosal in rats was inconclusive and reports on genotoxic effects in various in vitro systems were contradictory. Therefore, we reinvestigated thimerosal in the cytochalasin B block micronucleus test. Glutathione S-transferases were proposed to be involved in the detoxification of thimerosal or its decomposition products. Since the outcome of genotoxicity studies can be dependent on the metabolic competence of the cells used, we were additionally interested whether polymorphisms of glutathione S-transferases (GSTM1, GSTT1, or GSTP1) may influence the results of the micronucleus test with primary human lymphocytes. Blood samples of six healthy donors of different glutathione S-transferase genotypes were included in the study. At least two independent experiments were performed for each blood donor. Significant induction of micronuclei was seen at concentrations between 0.05-0.5 micro g/ml in 14 out of 16 experiments. Thus, genotoxic effects were seen even at concentrations which can occur at the injection site. Toxicity and toxicity-related elevation of micronuclei was seen at and above 0.6 micro g/ml thimerosal. Marked individual and intraindividual variations in the in vitro response to thimerosal among the different blood donors occurred. However, there was no association observed with any of the glutathione S-transferase polymorphism investigated. In conclusion, thimerosal is genotoxic in the cytochalasin B block micronucleus test with human lymphocytes. These data raise some concern on the widespread use of thimerosal.


An exploration of the knowledge and attitudes towards breastfeeding among a sample of Chinese mothers in Ireland.
Psychological factors are important in influencing breastfeeding practices. This retrospective study explored knowledge and attitudes related to breastfeeding of Chinese mothers living in Ireland. A cross-sectional self-administrated survey written in Chinese was distributed to a convenience sample of 322 immigrant Chinese mothers mainly via Chinese supermarkets and Chinese language schools in Dublin, with the involvement of the snowball method to increase sample size. Maternal breastfeeding knowledge and attitudes were described, their associations with socio-demographic variables were explored by Chi-square analysis, and their independent associations with breastfeeding behaviours were estimated by binary logistic regression analyses. In spite of considerable awareness of the advantages of breastfeeding (mean score = 4.03 ± 0.73), some misconceptions (e.g. 'mother should stop breastfeeding if she catches a cold') and negative attitudes (e.g. breastfeeding inconvenient, embarrassing, and adverse to mothers' figure) existed, especially among the less educated mothers. Cultural beliefs concerning the traditional Chinese postpartum diet were prevalent, particularly among those who had lived in Ireland for a shorter duration (P = 0.004). Psychological parameters had strong independent associations with breastfeeding practices in this study. Those who had lower awareness score (OR = 2.98, 95% CI: 1.87-4.73), more misconceptions and negative attitudes (P < 0.05), and weaker cultural beliefs (P < 0.05) were less likely to breastfeed. Findings highlight a need to focus resources and education on correcting the misconceptions identified and reversing the negative attitudes towards breastfeeding among Chinese mothers in Ireland, in particular those with primary/secondary level of education. Mothers' cultural beliefs should also be acknowledged and understood by healthcare providers.


Anomalous excitation spectra of frustrated quantum antiferromagnets.
We use series expansions to study the excitation spectra of spin-1/2 antiferromagnets on anisotropic triangular lattices. For the isotropic triangular lattice model (TLM), the high-energy spectra show several anomalous features that differ strongly from linear spin-wave theory (LSWT). Even in the Néel phase, the deviations from LSWT increase sharply with frustration, leading to rotonlike minima at special wave vectors. We argue that these results can be interpreted naturally in a spinon language and provide an explanation for the previously observed anomalous finite-temperature properties of the TLM. In the coupled-chains limit, quantum renormalizations strongly enhance the one-dimensionality of the spectra, in agreement with experiments on Cs2CuCl4.


Surgery for chronic aortic regurgitation: when should it be considered?
Deciding when to operate on a patient with chronic aortic regurgitation may be extremely difficult. The timing of surgery requires consideration of the etiology and pathophysiology of the aortic regurgitation, because aortic valve replacement carries morbidity and mortality that must be weighed against the potential problems of continued medical management. Guidelines for the use of surgery in patients with valvular disease have been developed by a joint task force of the American College of Cardiology and the American Heart Association. Practical recommendations based on these guidelines are presented.


Premature onset of degenerative disease of the cervical spine in rugby players.
Rugby players and other individuals with the changes of degenerative disease of the cervical spine are at risk of spinal cord trauma after hyperextension injury. In an attempt to assess whether rugby players are prone to the development of premature degenerative disease, radiographs of the cervical spines of 150 rugby players were compared with a control group of 150 male hospital patients. The study revealed that rugby players showed premature and advanced changes of degenerative disease when compared with the control group. These changes were most marked in the cervical spines of the tight forwards. Rugby players so affected are therefore more likely to present with the symptoms and signs of cervical osteo-arthrosis and are at greater risk of hyperextension injury to the cervical spinal cord.


Egalitarianism in female African lions.
Because most cooperative societies are despotic, it has been difficult to test models of egalitarianism. Female African lions demonstrate a unique form of plural breeding in which companions consistently produce similar numbers of surviving offspring. Consistent with theoretical predictions from models of reproductive skew, female lions are unable to control each other's reproduction because of high costs of fighting and low access to each other's newborn cubs. A female also lacks incentives to reduce her companions' reproduction, because her own survival and reproduction depend on group territoriality and synchronous breeding. Consequently, female relationships are highly symmetrical, and female lions are "free agents" who only contribute to communal care when they have cubs of their own.


Can symptoms of anosmia and dysgeusia be diagnostic for COVID-19?
Olfactory and taste dysfunction (OTD) is a potential neurological manifestation of coronavirus-2019 (COVID-19). We aimed to investigate the diagnostic value of symptoms of anosmia and dysgeusia for COVID-19. A comprehensive electronic search was conducted using PubMed, MEDLINE, Scopus, Cochrane database, and Google Scholar from 1 June 2020 to 12 June 2020. All studies reporting symptoms of anosmia and dysgeusia in COVID-19-positive patients were included. A total of 23 studies were included in the systematic review. Symptoms of anosmia and dysgeusia were frequently reported by COVID-19-positive patients. Symptoms were more common in females and in younger patients. There was no direct association between the severity of COVID-19 and the presence of symptoms. However, some evidence was found for a longer duration of these symptoms and increased severity of COVID-19 infection in young patients. OTD is commonly reported by COVID-19 patients. Due to limited literature on the association between OTD and COVID-19, it is currently not possible to conclude that these symptoms alone can be used to diagnose COVID-19. However, the presence of OTD can potentially be used as a screening tool for COVID-19 especially in young and female patients. Further research is required to establish the true diagnostic value of these symptoms and efficacy as screening tools for COVID-19 patients.


Who and what influences delayed presentation in breast cancer?
This study aimed to examine the extent and determinants of patient and general practitioner delay in the presentation of breast cancer. One hundred and eighty-five cancer patients attending a breast unit were interviewed 2 months after diagnosis. The main outcome measures were patient delay in presentation to the general practitioner and non-referral by the general practitioner to hospital after the patient's first visit. Nineteen per cent of patients delayed > or = 12 weeks. Patient delay was related to clinical tumour size > or = 4 cm (P = 0.0002) and with a higher incidence of locally advanced and metastatic disease (P = 0.01). A number of factors predicted patient delay: initial breast symptom(s) that did not include a lump (OR 4.5, P = 0.003), not disclosing discovery of the breast symptom immediately to someone else (OR 6.0, P < 0.001), seeking help only after being prompted by others (OR 4.4, P = 0.007) and presenting to the general practitioner with a non-breast problem (OR 3.5, P = 0.03). Eighty-three per cent of patients were referred to hospital directly after their first general practitioner visit. Presenting to the GP with a breast symptom that did not include a lump independently predicted general practitioner delay (OR 3.6, P = 0.002). In view of the increasing evidence that delay adversely affects survival, a large multicentre study is now warranted to confirm these findings that may have implications for public and medical education.


7 Tesla MRI in cerebral small vessel disease.
Cerebral small vessel disease (SVD) is a major cause of stroke and cognitive decline. Magnetic resonance imaging (MRI) currently plays a central role in diagnosis, and advanced MRI techniques are widely used in research but are limited by spatial resolution. Human 7 Tesla (7T) MRI has recently become available offering the ability to image at higher spatial resolution. This may provide additional insights into both the vascular pathology itself as well as parenchymal markers which could only previously be examined post mortem. In this review we cover the advantages and limitations of 7T MRI, review studies in SVD performed to date, and discuss potential future insights into SVD which 7T MRI may provide.


Drug-loaded nanocarriers: passive targeting and crossing of biological barriers.
Poor bioavailability and poor pharmacokinetic characteristics are some of the leading causes of drug development failure. Therefore, poorly-soluble drugs, fragile proteins or nucleic acid products may benefit from their encapsulation in nanosized vehicles, providing enhanced solubilization, protection against degradation, and increased access to pathological compartments. A key element for the success of drug-loaded nanocarriers is their ability to either cross biological barriers themselves, or allow loaded drugs to traverse them to achieve optimal pharmacological action at pathological sites. Depending on the mode of administration, nanocarriers may have to cross different physiological barriers in their journey towards their target. In this review, the crossing of biological barriers by passive targeting strategies will be presented for intravenous delivery (vascular endothelial lining, particularly for tumor vasculature and blood brain barrier targeting), oral administration (gastrointestinal lining), and upper airway administration (pulmonary epithelium). For each specific barrier, background information will be provided on the structure and biology of the tissues involved as well as available pathways for nano-objects or loaded drugs (diffusion and convection through fenestration, transcytosis, tight junction crossing, etc.). The determinants of passive targeting - size, shape, surface chemistry, surface patterning of nanovectors - will be discussed in light of current results. Perspectives on each mode of administration will be presented. The focus will be on polymeric nanoparticles and dendrimers, although advances in liposome technology will be also reported as they represent the largest body in the drug delivery literature.