The molecular characteristics of colorectal cancer: Impact of Ibuprofen and hyperthermia.
Despite various treatment options available for colorectal cancer, the survival rates for patients remain low. This study investigated the effects of hyperthermia and Ibuprofen on human colorectal adenocarcinoma cells (HT-29) viability, proliferation, and gene expression related to tumor suppression, Wnt signaling pathways, proliferation, and apoptosis The cells were exposed to hyperthermia at 42 or 43°C for 3 hours or Ibuprofen at different concentrations (700-1500 μM), and the effects were analyzed through MTT assay, trypan blue staining, and quantitative Real-time PCR. The study used quantitative Real-time PCR (qRT-PCR) to evaluate the effect of hyperthermia and Ibuprofen on the expression of various genes associated with tumor suppression, proliferation, Wnt signaling pathway, and apoptosis. The results revealed that hyperthermia caused a minor reduction in the viability and proliferation of HT-29 cells, but the decrease was not statistically significant (P<0.05). On the other hand, Ibuprofen caused a concentration-dependent decrease in the viability and proliferation of HT-29 cells. Both hyperthermia and Ibuprofen reduced the expression of WNT1, CTNNB1, BCL2, and PCNA genes, and increased the expression of KLF4, P53, and BAX genes. However, the changes in gene expression were not statistically significant in cells treated with hyperthermia. The findings suggest that Ibuprofen is more effective in reducing cancer cell proliferation by promoting apoptosis and inhibiting the Wnt signaling pathway than hyperthermia, which had some impact but was not statistically significant. The study highlights the potential of Ibuprofen as a targeted therapy for colorectal cancer.


Somatic mosaicism in Menkes disease suggests choroid plexus-mediated copper transport to the developing brain.
The primary mechanism of copper transport to the brain is unknown, although this process is drastically impaired in Menkes disease, an X-linked neurodevelopmental disorder caused by mutations in an evolutionarily conserved copper transporter, ATP7A. Potential central nervous system entry routes for copper include brain capillary endothelial cells that originate from mesodermal angioblasts and form the blood-brain barrier, and the choroid plexuses, which derive from embryonic ectoderm, and form the blood-cerebrospinal fluid barrier. We exploited a rare (and first reported) example of somatic mosaicism for an ATP7A mutation to shed light on questions about copper transport into the developing brain. In a 20-month-old Menkes disease patient evaluated before copper treatment, blood copper, and catecholamine concentrations were normal, whereas levels in cerebrospinal fluid were abnormal and consistent with his neurologically severe phenotype. We documented disparate levels of mosaicism for an ATP7A missense mutation, P1001L, in tissues derived from different embryonic origins; allele quantitation showed P1001L in approximately 27% of DNA samples from blood cells (mesoderm-derived) and 88% from cultured fibroblasts (ectoderm-derived). These findings imply that the P1001L mutation in the patient preceded formation of the three primary embryonic lineages at gastrulation, with the ectoderm layer ultimately harboring a higher percentage of mutation-bearing cells than mesoderm or endoderm. Since choroid plexus epithelia are derived from neuroectoderm, and brain capillary endothelial cells from mesodermal angioblasts, the clinical and biochemical findings in this infant support a critical role for the blood-CSF barrier (choroid plexus epithelia) in copper entry to the developing brain.


Observational Dutch Young Symptomatic StrokE studY (ODYSSEY): study rationale and protocol of a multicentre prospective cohort study.
The proportion of strokes occurring in younger adults has been rising over the past decade. Due to the far longer life expectancy in the young, stroke in this group has an even larger socio-economic impact. However, information on etiology and prognosis remains scarce. ODYSSEY is a multicentre prospective cohort study on the prognosis and risk factors of patients with a first-ever TIA, ischemic stroke or intracerebral hemorrhage aged 18 to 49 years. Our aim is to include 1500 patients. Primary outcome will be all cause mortality and risk of recurrent vascular events. Secondary outcome will be the risk of post-stroke epilepsy and cognitive impairment. Patients will complete structured questionnaires on outcome measures and risk factors. Both well-documented and less well-documented risk factors and potentially acute trigger factors will be investigated. Patients will be followed every 6 months for at least 3 years. In addition, an extensive neuropsychological assessment will be administered both at baseline and 1 year after the stroke/TIA. Furthermore we will include 250 stroke-free controls, who will complete baseline assessment and one neuropsychological assessment. ODYSSEY is designed to prospectively determine prognosis after a young stroke and get more insight into etiology of patients with a TIA, ischemic stroke and intracerebral hemorrhage in patients aged 18 to 49 years old in a large sample size.


Axillorenal arteriovenous graft: a new approach for dialysis access.
As the population requiring hemodialysis grows, it becomes increasingly common to encounter patients with limited options for vascular access. Because inability to secure vascular access is a life-threatening problem, it is important to consider all possible options in each patient. We report a new arteriovenous grafting procedure in which the left renal vein is used for outflow in a patient with multiple venous occlusions. Patency of the graft continues 18 months after placement. This graft carries acceptable morbidity, and can be revised. Consideration of this graft is appropriate in selected patients.


Managing boundaries in primary care service improvement: a developmental approach to communities of practice.
Effective implementation of change in healthcare organisations involves multiple professional and organisational groups and is often impeded by professional and organisational boundaries that present relatively impermeable barriers to sharing knowledge and spreading work practices. Informed by the theory of communities of practice (CoPs), this study explored the effects of intra-organisational and inter-organisational boundaries on the implementation of service improvement within and across primary healthcare settings and on the development of multiprofessional and multi-organisational CoPs during this process. The study was conducted within the Collaboration for Leadership in Applied Health Research and Care (CLAHRC) for Greater Manchester-a collaborative partnership between the University of Manchester and local National Health Service organisations aiming to undertake applied health research and enhance its implementation in clinical practice. It deployed a qualitative embedded case study design, encompassing semistructured interviews, direct observation and documentary analysis, conducted in 2010-2011. The sample included practice doctors, nurses, managers and members of the CLAHRC implementation team. The study showed that in spite of epistemic and status differences, professional boundaries between general practitioners, practice nurses and practice managers co-located in the same practice over a relatively long period of time could be successfully bridged, leading to the formation of multiprofessional CoPs. While knowledge circulated relatively easily within these CoPs, barriers to knowledge sharing emerged at the boundary separating them from other groups existing in the same primary care setting. The strongest boundaries, however, lay between individual general practices, with inter-organisational knowledge sharing and collaboration between them remaining unequally developed across different areas due to historical factors, competition and strong organisational identification. Manipulated emergence of multi-organisational CoPs in the context of primary care may thus be problematic. In cases when manipulated emergence of new CoPs is problematic, boundary issues could be addressed by adopting a developmental perspective on CoPs, which provides an alternative to the analytical and instrumental perspectives previously described in the CoP literature. This perspective implies a pragmatic, situational approach to mapping existing CoPs and their characteristics and potentially modifying them in the process of service improvement through the combination of internal and external facilitation.


Glucose-induced changes in the phenotype of human peritoneal mesothelial cells: effect of L-2-oxothiazolidine carboxylic acid.
During peritoneal dialysis, mesothelial cells are chronically exposed to high concentrations of glucose. Therefore, the cytotoxic effect of glucose may alter the function and reactivity of these cells. For 4 weeks, human peritoneal mesothelial cells were cultured in vitroin medium supplemented with 45 mM glucose or 45 mM mannitol or with 45 mM glucose and 1 mM L-2-oxothiazolidine-4-carboxylic acid (OTZ), the latter being a precursor for glutathione synthesis. Peroxidation of the mesothelial cell lipids, synthetic activity and reaction of these cells to peritoneal dialysis fluids were studied. In contrast to mannitol, glucose enhanced the peroxidation of the cellular lipids (+65%, p < 0.01) an effect that was prevented by OTZ. Synthesis of hyaluronan and vascular endothelial growth factor was reduced in mesothelial cells treated with glucose by 36% (p < 0.01) and 44% (p < 0.05), respectively; both glucose effects were reversed when cells were incubated with glucose plus OTZ. Monocyte chemoattractant protein-1 synthesis by cells exposed to glucose was increased by 31% (p < 001), and again that effect was prevented by OTZ. Glucose and mannitol stimulated synthesis of fibronectin (+32%, p < 0.05). Mesothelial cells chronically exposed to glucose became activated after subsequent exposure to the dialysis fluid, as reflected by the increased release of interleukin (IL)-6, in contrast to control mesothelial cells, in which IL-6 synthesis was suppressed. Chronic exposure of mesothelial cells to glucose changes their synthetic activity and their reaction after exposure to dialysis fluids. Some of these effects are prevented by OTZ, which suggests that glucose-induced free radicals are responsible for a change in mesothelial cell phenotype under the conditions of peritoneal dialysis.


First Report of the Armillaria Root Disease Pathogen, Armillaria gallica, on Douglas-fir (Pseudotsuga menziesii) in Arizona.
In August 2010, a mycelial fan (isolate AZ32F) of Armillaria sp. was collected from the root collar of a living Douglas-fir tree on the Mogollon Rim within the Coconino National Forest (approximate location 34°25'31.26″N, 111°20'41.04″W, elevation 2,293 m) in central Arizona. Mycelial fans under the bark of living trees are a sign of pathogenicity, and symptoms of the diseased tree included resinosis, sloughing bark, and thinning crown. The infected tree was located on a south-facing slope with approximately 30% tree cover, dominated by ponderosa pine (Pinus ponderosa), with lesser components of Douglas-fir and Gambel oak (Quercus gambelii). Based on three replications of somatic incompatibility tests against 24 tester isolates representing seven North American Armillaria spp., isolate AZ32F showed 100% intraspecific compatibility (colorless antagonism) with all four A. gallica isolates, 22% compatibility with A. calvescens, and 0% compatibility with the remaining Armillaria spp. Based on GenBank BLASTn of isolate AZ32F sequences, the partial LSU-IGS1 (GenBank Accession No. KF186682) showed 99 to 100% similarity to A. gallica and two other related Armillaria spp. with 99 to 100% coverage, and translation elongation factor-1 alpha (tef-1α) sequences (KC525954) showed 96% similarity to A. gallica (JF895844) with 100% coverage. Thus, isolate AZ32F was identified as A. gallica, based on somatic incompatibility tests and DNA sequences (partial LSU-IGS1 and tef-1α). Although the isolate is identified as A. gallica with similarities to other North American isolates, evidence is mounting that currently recognized A. gallica likely represents a species complex that comprises multiple phylogenetic species (4). Previous surveys in Arizona have noted A. mellea and A. solidipes (as A. ostoyae) (3), but A. gallica has never been previously confirmed in this state. Within North America, A. gallica is commonly reported east of the Rocky Mountains and in West Coast states of the United States, where it infects hardwoods and conifers including Douglas-fir (1,2). Its ecological behavior ranges from saprophyte to weak/aggressive pathogen (1,2). Because damage by A. gallica appears to increase on hosts predisposed by stress (1), further surveys are needed to document its distribution, frequency, and ecological behavior in the southwestern United States, where climate change will likely cause tree stress due to maladaptation. Continued surveys for Armillaria spp. will better determine their potential threat within the geologically and ecologically unique Mogollon Rim of Arizona. References: (1) K. Baumgartner and D. M. Rizzo. Plant Dis. 85:947, 2001. (2) N. J. Brazee and R. L. Wick. For. Ecol. Manage. 258:1605, 2009. (3) R. L. Gilbertson and D. M. Bigelow. J. Arizona-Nevada Acad. Sci. 31:13, 1998. (4) M.-S. Kim et al. Phytopathology 102:S4.63, 2012.


Perception and modification of plant flavonoid signals by rhizosphere microorganisms.
Flavonoids are a diverse class of polyphenolic compounds that are produced as a result of plant secondary metabolism. They are known to play a multifunctional role in rhizospheric plant-microbe and plant-plant communication. Most familiar is their function as a signal in initiation of the legume-rhizobia symbiosis, but, flavonoids may also be signals in the establishment of arbuscular mycorrhizal symbiosis and are known agents in plant defence and in allelopathic interactions. Flavonoid perception by, and impact on, their microbial targets (e.g. rhizobia, plant pathogens) is relatively well characterized. However, potential impacts on 'non-target' rhizosphere inhabitants ('non-target' is used to distinguish those microorganisms not conventionally known as targets) have not been thoroughly investigated. Thus, this review first summarizes the conventional roles of flavonoids as nod gene inducers, phytoalexins and allelochemicals before exploring questions concerning 'non-target' impacts. We hypothesize that flavonoids act to shape rhizosphere microbial community structure because they represent a potential source of carbon and toxicity and that they impact on rhizosphere function, for example, by accelerating the biodegradation of xenobiotics. We also examine the reverse question, 'how do rhizosphere microbial communities impact on flavonoid signals?' The presence of microorganisms undoubtedly influences the quality and quantity of flavonoids present in the rhizosphere, both through modification of root exudation patterns and microbial catabolism of exudates. Microbial alteration and attenuation of flavonoid signals may have ecological consequences for below-ground plant-microbe and plant-plant interaction. We have a lack of knowledge concerning the composition, concentration and bioavailability of flavonoids actually experienced by microbes in an intact rhizosphere, but this may be addressed through advances in microspectroscopic and biosensor techniques. Through the use of plant mutants defective in flavonoid biosynthesis, we may also start to address the question of the significance of flavonoids in shaping rhizosphere community structure and function.


Substitution of murine ferrochelatase glutamate-287 with glutamine or alanine leads to porphyrin substrate-bound variants.
Ferrochelatase (EC 188.8.131.52) is the terminal enzyme of the haem biosynthetic pathway and catalyses iron chelation into the protoporphyrin IX ring. Glutamate-287 (E287) of murine mature ferrochelatase is a conserved residue in all known sequences of ferrochelatase, is present at the active site of the enzyme, as inferred from the Bacillus subtilis ferrochelatase three-dimensional structure, and is critical for enzyme activity. Substitution of E287 with either glutamine (Q) or alanine (A) yielded variants with lower enzymic activity than that of the wild-type ferrochelatase and with different absorption spectra from the wild-type enzyme. In contrast to the wild-type enzyme, the absorption spectra of the variants indicate that these enzymes, as purified, contain protoporphyrin IX. Identification and quantification of the porphyrin bound to the E287-directed variants indicate that approx. 80% of the total porphyrin corresponds to protoporphyrin IX. Significantly, rapid stopped-flow experiments of the E287A and E287Q variants demonstrate that reaction with Zn(2+) results in the formation of bound Zn-protoporphyrin IX, indicating that the endogenously bound protoporphyrin IX can be used as a substrate. Taken together, these findings suggest that the structural strain imposed by ferrochelatase on the porphyrin substrate as a critical step in the enzyme catalytic mechanism is also accomplished by the E287A and E287Q variants, but without the release of the product. Thus E287 in murine ferrochelatase appears to be critical for the catalytic process by controlling the release of the product.


Virtual Reality Experiments with Physiological Measures.
Virtual reality (VR) experiments are increasingly employed because of their internal and external validity compared to real-world observation and laboratory experiments, respectively. VR is especially useful for geographic visualizations and investigations of spatial behavior. In spatial behavior research, VR provides a platform for studying the relationship between navigation and physiological measures (e.g., skin conductance, heart rate, blood pressure). Specifically, physiological measures allow researchers to address novel questions and constrain previous theories of spatial abilities, strategies, and performance. For example, individual differences in navigation performance may be explained by the extent to which changes in arousal mediate the effects of task difficulty. However, the complexities in the design and implementation of VR experiments can distract experimenters from their primary research goals and introduce irregularities in data collection and analysis. To address these challenges, the Experiments in Virtual Environments (EVE) framework includes standardized modules such as participant training with the control interface, data collection using questionnaires, the synchronization of physiological measurements, and data storage. EVE also provides the necessary infrastructure for data management, visualization, and evaluation. The present paper describes a protocol that employs the EVE framework to conduct navigation experiments in VR with physiological sensors. The protocol lists the steps necessary for recruiting participants, attaching the physiological sensors, administering the experiment using EVE, and assessing the collected data with EVE evaluation tools. Overall, this protocol will facilitate future research by streamlining the design and implementation of VR experiments with physiological sensors.


Abortion surveillance - United States, 2011.
Since 1969, CDC has conducted abortion surveillance to document the number and characteristics of women obtaining legal induced abortions in the United States. 2011. Each year, CDC requests abortion data from the central health agencies of 52 reporting areas (the 50 states, the District of Columbia, and New York City). The reporting areas provide this information voluntarily. For 2011, data were received from 49 reporting areas. For trend analysis, abortion data were evaluated from 46 areas that reported data every year during 2002-2011. Census and natality data, respectively, were used to calculate abortion rates (number of abortions per 1,000 women) and ratios (number of abortions per 1,000 live births). A total of 730,322 abortions were reported to CDC for 2011. Of these abortions, 98.3% were from the 46 reporting areas that provided data every year during 2002-2011. Among these same 46 reporting areas, the abortion rate for 2011 was 13.9 abortions per 1,000 women aged 15-44 years, and the abortion ratio was 219 abortions per 1,000 live births. From 2010 to 2011, the total number and rate of reported abortions decreased 5% and the abortion ratio decreased 4%, and from 2002 to 2011, the total number, rate, and ratio of reported abortions decreased 13%, 14%, and 12%, respectively. In 2011, all three measures reached their lowest level for the entire period of analysis (2002-2011). In 2011 and throughout the period of analysis, women in their 20s accounted for the majority of abortions and had the highest abortion rates, and women in their 30s and older accounted for a much smaller percentage of abortions and had lower abortion rates. In 2011, women aged 20-24 and 25-29 years accounted for 32.9% and 24.9% of all abortions, respectively, and had abortion rates of 24.9 and 19.4 abortions per 1,000 women aged 20-24 and 25-29 years, respectively. In contrast, women aged 30-34, 35-39, and ≥40 years accounted for 15.8%, 8.9%, and 3.6% of all abortions, respectively, and had abortion rates of 12.7, 7.5, and 2.8 abortions per 1,000 women aged 30-34 years, 35-39 years, and ≥40 years, respectively. Throughout the period of analysis, abortion rates decreased among women aged 20-24 and 25-29 years by 21% and 16%, respectively, whereas they increased among women aged ≥40 years by 8%. In 2011, adolescents aged <15 and 15-19 years accounted for 0.4% and 13.5% of all abortions, respectively, and had abortion rates of 0.9 and 10.5 abortions per 1,000 adolescents aged <15 and 15-19 years, respectively. From 2002 to 2011, the percentage of abortions accounted for by adolescents aged 15-19 years decreased 21% and their abortion rate decreased 34%. These decreases were greater than the decreases for women in any older age group. In contrast to the percentage distribution of abortions and abortion rates by age, abortion ratios in 2011 and throughout the entire period of analysis were highest among adolescents and lowest among women aged 30-39 years. Abortion ratios decreased from 2002 to 2011 for women in all age groups except for those aged <15 years, for whom they increased. In 2011, most (64.5%) abortions were performed by ≤8 weeks' gestation, and nearly all (91.4%) were performed by ≤13 weeks' gestation. Few abortions (7.3%) were performed between 14-20 weeks' gestation or at ≥21 weeks' gestation (1.4%). From 2002 to 2011, the percentage of all abortions performed at ≤8 weeks' gestation increased 6%. In 2011, among reporting areas that included medical (nonsurgical) abortion on their reporting form, a total of 71.0% of abortions were performed by curettage at ≤13 weeks' gestation, 19.1% were performed by early medical abortion (a nonsurgical abortion at ≤8 weeks' gestation), and 8.6% were performed by curettage at >13 weeks' gestation; all other methods were uncommon. Among abortions performed at ≤8 weeks' gestation that were eligible for early medical abortion on the basis of gestational age, 28.5% were completed by this method. The percentage of abortions reported as early medical abortions increased 3% from 2010 to 2011. Deaths of women associated with complications from abortions for 2011 are being investigated as part of CDC's Pregnancy Mortality Surveillance System. In 2010, the most recent year for which data were available, 10 women were identified to have died as a result of complications from known legal induced abortions. No reported deaths were associated with known illegal induced abortions. Among the 46 areas that reported data every year during 2002-2011, large decreases in the total number, rate, and ratio of reported abortions from 2010 to 2011, in combination with decreases that occurred during 2008-2010, resulted in historic lows for all three measures of abortion. Unintended pregnancy is the major contributor to abortion. Because unintended pregnancies are rare among women who use the most effective methods of contraception, increasing access to and use of these methods can help further reduce the number of abortions performed in the United States. The data in this report can help program planners and policy makers identify groups of women at greatest risk for unintended pregnancy and help guide and evaluate prevention efforts.


Impact of alternative management policies on health-related quality of life in women with low-grade abnormal cervical cytology referred for colposcopy.
To assess the health-related quality of life (HRQoL) over 30 months among women who received colposcopy following low-grade abnormal cervical cytology. 751 women with low-grade abnormal cytology were seen for colposcopy. Of these, 350 additionally underwent either immediate treatment by large loop excision of the transformation zone (LLETZ) or investigation by punch biopsy followed by treatment of high-grade cervical intraepithelial neoplasia if present. The HRQoL of the women was assessed on seven separate occasions over 30 months by means of the EQ-5D instrument. Outcomes for women receiving colposcopy only, colposcopy with biopsy and colposcopy with LLETZ were compared. Women experienced modest short-term increases in HRQoL, statistically significant for the colposcopy only and biopsy groups but not for the LLETZ group. HRQoL in all three groups thereafter fell until 12 months following recruitment, significantly so for the LLETZ group. Changes in EQ-5D index score arose primarily from changes in severities in the "pain and discomfort" and "anxiety and depression" domains. Changes in the visual analogue scale (VAS) representations of quality of life corresponded closely to those of the index scores and most of the VAS scores themselves did not differ from VAS population norms. All inter-assessment changes in index scores were smaller than the minimum important difference for the instrument. Beyond 18 months from recruitment, HRQoL stabilised in all three groups. Women referred to colposcopy following a low-grade abnormal smear test result experienced a short-term improvement in their health-related quality of life, but the long-term effect was insubstantial. HRQoL over the post-recruitment follow-up period did not vary by intervention.


In vivo measurements of limbic glutamate and GABA concentrations in epileptic patients during affective and cognitive tasks: A microdialysis study.
Limbic system structures such as the amygdala (AMG) and the hippocampus (HIPP) are involved in affective and cognitive processing. However, because of the limitations in noninvasive technology, absolute concentrations of the neurotransmitters underlying limbic system engagement are not known. Here, we report changes in the concentrations of the neurotransmitters glutamate (Glu) and gamma-aminobutyric acid (GABA) in the HIPP and the AMG of patients with nonlesional temporal lobe epilepsy undergoing surgery for intracranial subdural and depth electrode implantation. We utilized an in-vivo microdialysis technique while subjects were engaged in cognitive tasks with or without emotional content. The performance of an emotion learning task (EmoLearn) was associated with a significant increase in the concentration of glutamate in the HIPP when images with high valence content were processed, as compared to its concentration while processing images with low valence. In addition, significantly decreased levels of glutamate were found in the AMG when images with predominantly low valence content were processed, as compared to its concentration at baseline. The processing of face stimuli with anger/fear content (FaceMatch task) was accompanied with significantly decreased concentrations of GABA in the AMG and HIPP compared to its levels at the baseline. The processing of shapes on the other hand was accompanied with a significantly decreased concentration of the glutamate in the AMG as well as in the HIPP compared to the baseline. Finally, the performance of a nondeclarative memory task (weather prediction task-WPT) was associated with relatively large and opposite changes in the GABA levels compared to the baseline in the AMG (decrease) and the HIPP (increase). These data are relevant for showing an involvement of the amygdala and the hippocampus in emotional processing and provide additional neurochemical clues towards a more refined model of the functional circuitry of the human limbic system.


Proteomic analysis of the anti-inflammatory action of minocycline.
Minocycline possesses anti-inflammatory properties independently of its antibiotic activity although the underlying molecular mechanisms are unclear. Lipopolysaccharide (LPS)-induced cytokines and pro-inflammatory protein expression are reduced by minocycline in cultured macrophages. Here, we tested a range of clinically important tetracycline compounds (oxytetracycline, doxycycline, minocycline and tigecycline) and showed that they all inhibited LPS-induced nitric oxide production. We made the novel finding that tigecycline inhibited LPS-induced nitric oxide production to a greater extent than the other tetracycline compounds tested. To identify potential targets for minocycline, we assessed alterations in the macrophage proteome induced by LPS in the presence or absence of a minocycline pre-treatment using 2-DE and nanoLC-MS. We found a number of proteins, mainly involved in cellular metabolism (ATP synthase β-subunit and aldose reductase) or stress response (heat shock proteins), which were altered in expression in response to LPS, some of which were restored, at least in part, by minocycline. This is the first study to document proteomic changes induced by minocycline. The observation that minocycline inhibits some, but not all, of the LPS-induced proteomic changes shows that minocycline specifically affects some signalling pathways and does not completely inhibit macrophage activation.


Understanding pitch perception as a hierarchical process with top-down modulation.
Pitch is one of the most important features of natural sounds, underlying the perception of melody in music and prosody in speech. However, the temporal dynamics of pitch processing are still poorly understood. Previous studies suggest that the auditory system uses a wide range of time scales to integrate pitch-related information and that the effective integration time is both task- and stimulus-dependent. None of the existing models of pitch processing can account for such task- and stimulus-dependent variations in processing time scales. This study presents an idealized neurocomputational model, which provides a unified account of the multiple time scales observed in pitch perception. The model is evaluated using a range of perceptual studies, which have not previously been accounted for by a single model, and new results from a neurophysiological experiment. In contrast to other approaches, the current model contains a hierarchy of integration stages and uses feedback to adapt the effective time scales of processing at each stage in response to changes in the input stimulus. The model has features in common with a hierarchical generative process and suggests a key role for efferent connections from central to sub-cortical areas in controlling the temporal dynamics of pitch processing.


TarO: a target optimisation system for structural biology.
TarO (http://www.compbio.dundee.ac.uk/taro) offers a single point of reference for key bioinformatics analyses relevant to selecting proteins or domains for study by structural biology techniques. The protein sequence is analysed by 17 algorithms and compared to 8 databases. TarO gathers putative homologues, including orthologues, and then obtains predictions of properties for these sequences including crystallisation propensity, protein disorder and post-translational modifications. Analyses are run on a high-performance computing cluster, the results integrated, stored in a database and accessed through a web-based user interface. Output is in tabulated format and in the form of an annotated multiple sequence alignment (MSA) that may be edited interactively in the program Jalview. TarO also simplifies the gathering of additional annotations via the Distributed Annotation System, both from the MSA in Jalview and through links to Dasty2. Routes to other information gateways are included, for example to relevant pages from UniProt, COG and the Conserved Domains Database. Open access to TarO is available from a guest account with private accounts for academic use available on request. Future development of TarO will include further analysis steps and integration with the Protein Information Management System (PIMS), a sister project in the BBSRC 'Structural Proteomics of Rational Targets' initiative.


Quality Assurance in Telehealth: Adherence to Evidence-Based Indicators.
Background: Value enhancing telehealth (TH) lacks a robust body of formal clinically focused quality assessment studies. Innovations such as telehealth must always demonstrate that it preserves or hopefully advances quality. Introduction: We sought to determine whether adherence to the evidence-based Choosing Wisely (CW) recommendations (antibiotic stewardship) for acute sinusitis differs for encounters through direct-to-consumer (DTC) telemedicine verses "in-person" care in an emergency department (ED) or an urgent care (UC) center. Materials and Methods: Study design was a retrospective review. Patients with a symptom complex consistent with acute sinusitis treated through DTC were matched with ED and UC patients, based upon time of visit. Charts were reviewed to determine patient characteristics, chief complaint, final diagnosis, presence or absence of criteria within the CW guidelines, and whether or not antibiotics were prescribed. The main outcome was adherence to the CW campaign recommendations. Results: A total of 570 visits were studied: 190 DTC, 190 ED, and 190 UC visits. The predominant chief complaints were upper respiratory infection (36%), sore throat (25%), and sinusitis (18%). Overall, there was a 67% (95% CI 62.3-71.7) adherence rate with the CW guidelines for sinusitis: DTC visits (71%), ED visits (68%), and UC visits (61%). There was a nonsignificant difference (p = 0.29) in adherence to CW guidelines based upon type of visit (DTC, UC, and ED). Discussion: The challenge is to demonstrate whether or not DTC TH compromises quality. Conclusion: In this study, DTC visits were associated with at least as good an adherence to the CW campaign recommendations as emergency medicine (EM) and UC in-person visits.


In vivo reactivation of latent herpes simplex virus 1 in mice can occur in the brain before occurring in the trigeminal ganglion.
Herpes simplex virus 1 (HSV-1) establishes latency in neurons of the brains and sensory ganglia of humans and experimentally infected mice. The latent virus can reactivate to cause recurrent infection. Both primary and recurrent infections can induce diseases, such as encephalitis. In humans, the majority of encephalitis cases occur as a recurrent infection. However, in the past, numerous mouse studies documented that viral reactivation occurs efficiently in the ganglion, but extremely rarely in the brain, when assessed ex vivo by cultivating minced tissue explants. Here, we compare the brains and the trigeminal ganglia of mice latently infected with HSV-1 (strain 294.1 or McKrae) for levels of viral genomes and in vivo reactivation. The numbers of copies of 294.1 and McKrae genomes in the brain stem were significantly greater than those in the trigeminal ganglion. Most importantly, 294.1 and McKrae reactivation was detected in the brain stems earlier than in the trigeminal ganglia of mice treated with hyperthermia to reactivate latent virus in vivo. In addition, the brain stem yielded reactivated virus at a high frequency compared with the trigeminal ganglion, especially in mice latently infected with 294.1 after hyperthermia treatment. These results provide evidence that recurrent brain infection can be induced by the reactivation of latent virus in the brain in situ. Herpes simplex virus 1 (HSV-1) establishes latency in neurons of the brains and sensory ganglia of humans and experimentally infected mice. The latent virus can reactivate to cause recurrent infection. In the past, studies of viral reactivation focused on the ganglion, because efficient viral reactivation was detected in the ganglion but not in the brain when assessed ex vivo by cultivating mouse tissue explants. In this study, we report that the brain contains more viral genomes than the trigeminal ganglion in latently infected mice. Notably, the brain yields reactivated virus early and efficiently compared with the trigeminal ganglion after mice are stimulated to reactivate latent virus. Our findings raise the potential importance of HSV-1 latent infection and reactivation in the brain.


[Scuba diving: barotrauma, decompression sickness, pulmonary contra-indications].
The practice of scuba diving is associated with two specific medical problems: barotrauma directly related to changes in ambient pressure, and decompression sickness related to the uptake and the release of inert gases by the body. Neurological symptoms are frequent in severe diving accidents. They may arise following either barotrauma or decompression sickness, and often require urgent treatment in a hyperbaric chamber. Asthma, chronic obstructive pulmonary disease, and spontaneous pneumothorax increase the risk of lung barotrauma and represent contraindications to diving.


Solution structure of eggcase silk protein and its implications for silk fiber formation.
Spider silks are renowned for their excellent mechanical properties and biomimetic and industrial potentials. They are formed from the natural refolding of water-soluble fibroins with alpha-helical and random coil structures in silk glands into insoluble fibers with mainly beta-structures. The structures of the fibroins at atomic resolution and silk formation mechanism remain largely unknown. Here, we report the 3D structures of individual domains of a approximately 366-kDa eggcase silk protein that consists of 20 identical type 1 repetitive domains, one type 2 repetitive domain, and conserved nonrepetitive N- and C-terminal domains. The structures of the individual domains in solution were determined by using NMR techniques. The domain interactions were investigated by NMR and dynamic light-scattering techniques. The formation of micelles and macroscopic fibers from the domains was examined by electron microscopy. We find that either of the terminal domains covalently linked with at least one repetitive domain spontaneously forms micelle-like structures and can be further transformed into fibers at > or = 37 degrees C and a protein concentration of > 0.1 wt%. Our biophysical and biochemical experiments indicate that the less hydrophilic terminal domains initiate the assembly of the proteins and form the outer layer of the micelles whereas the more hydrophilic repetitive domains are embedded inside to ensure the formation of the micelle-like structures that are the essential intermediates in silk formation. Our results establish the roles of individual silk protein domains in fiber formation and provide the basis for designing miniature fibroins for producing artificial silks.


Cervical spondylomyelopathy (wobbler syndrome) in dogs.
Cervical spondylomyelopathy (CSM) is a common and controversial disease of the cervical spine of large and giant breed dogs. Not many diseases in veterinary medicine have been referred to by 14 different names and have had 21 surgical techniques proposed to treat it. This article reviews the current knowledge of CSM with regard to its etiology, pathophysiology, diagnosis, treatment, and outcome.


Measuring the α-particle charge radius with muonic helium-4 ions.
The energy levels of hydrogen-like atomic systems can be calculated with great precision. Starting from their quantum mechanical solution, they have been refined over the years to include the electron spin, the relativistic and quantum field effects, and tiny energy shifts related to the complex structure of the nucleus. These energy shifts caused by the nuclear structure are vastly magnified in hydrogen-like systems formed by a negative muon and a nucleus, so spectroscopy of these muonic ions can be used to investigate the nuclear structure with high precision. Here we present the measurement of two 2S-2P transitions in the muonic helium-4 ion that yields a precise determination of the root-mean-square charge radius of the α particle of 1.67824(83) femtometres. This determination from atomic spectroscopy is in excellent agreement with the value from electron scattering1, but a factor of 4.8 more precise, providing a benchmark for few-nucleon theories, lattice quantum chromodynamics and electron scattering. This agreement also constrains several beyond-standard-model theories proposed to explain the proton-radius puzzle2-5, in line with recent determinations of the proton charge radius6-9, and establishes spectroscopy of light muonic atoms and ions as a precise tool for studies of nuclear properties.


Hidradenitis Suppurativa and Bipolar Disorders: A Population-Based Study.
Psychological comorbidities have been reported in patients with hidradenitis suppurativa (HS), yet only a few studies have investigated the possible association between HS and severe psychiatric disorders. The current study aimed to assess the association between HS and bipolar disorder, a major, chronic, psychiatric disease. A nationwide, population-based study was performed utilizing data from the Clalit Health Services (CHS) database in Israel. The study included 4,191 HS patients and 20,941 age- and gender-matched controls. A multivariate binary logistic regression, adjusted for demographic and behavioral risk factors, was performed. A higher proportion of bipolar disorders was found among HS patients compared to controls (0.7 vs. 0.1%, respectively). There was an increased proportion of active smokers among HS patients than among controls (53.4 vs. 13.5%, respectively). In the multivariate analysis, adjusting for age, sex, and smoking, HS was found to be independently and positively associated with bipolar disorders (OR 2.12, 95% CI 1.21-3.27, p < 0.01), yet after controlling for body mass this association became nonsignificant. Bipolar disorders are associated with HS. Future studies should explore whether cardiometabolic deficiencies might account for this association. Healthcare providers should consider this potential co-occurrence as it may impede patient compliance and require appropriate screening and treatment. Results also stress the need for a multidisciplinary approach to optimize management of the disease and its associated comorbidities.


Vestibular and balance rehabilitation therapy.
A prospective observational study of the performance of patients enrolled in an individually customized program of habituation and balance rehabilitation physical therapy was conducted from January 1988 to January 1990. Patients ranged in age from 20 to 89 years, with a wide variety of diagnoses. Two global outcome measures--posttherapy symptom response score and pretherapy and posttherapy disability score--were developed and used to judge overall patient performance. In addition, two specific indicators--one for balance performance (dynamic posturography) and one for sensitivity to rapid head movements (motion sensitivity quotient)--were used to measure performance in these two areas. Results indicate statistically significant changes before versus after therapy for both specific measures, and 80% to 85% of the patients showed a reduction in symptoms and disability score following therapy. Analysis of variance and multiple regression analysis indicate that nature of symptoms, pretherapy disability level, history of head injury, and results on dynamic posturography were the variables most predictive of therapy outcome, while age and duration of symptoms made no difference. Other variables, such as medications and site of lesion, were found to affect the length of therapy but not the outcome.


Health effects of phytoestrogens.
Phytoestrogens are naturally occurring plant-derived phytochemicals, whose common biological roles are to protect plants from stress or to act as part of a plant's defense mechanism. Although composed of a wide group of nonsteroidal compounds of diverse structure, phytoestrogens have been shown to bind estrogen receptors and to behave as weak agonist/antagonist in both animals and humans. Phytoestrogens include mainly isoflavones (IF), coumestans, and lignans. These compounds are known to be present in fruits, vegetables, and whole grains commonly consumed by humans. IF are found in legumes--mainly soybeans--whereas flaxseed is a major source of lignans, and coumestans are significantly present in clover, alfalfa and soybean sprouts. 8-Prenyl flavonoids are common in vegetables. Bioavailability of IF requires an initial hydrolysis of the sugar moiety by intestinal beta-glucosidases to allow the following uptake by enterocytes and the flow through the peripheral circulation. Following absorption, IF are then reconjugated mainly to glucuronic acid and to a lesser degree to sulphuric acid. Gut metabolism seems key to the determination of the potency of action. Several epidemiological studies correlated high dose consumptions of soy IF with multiple beneficial effects on breast and prostate cancers, menopausal symptoms, osteoporosis, atherosclerosis and stroke, and neurodegeneration. For the relief of menopausal symptoms a consumption of 60 mg aglycones/day has been suggested; for cancer prevention a consumption between 50 and 110 mg aglycones/day is considered beneficial to reduce risks of breast, colon and prostate cancer; to decrease cardiovascular risk a minimum intake of 40-60 mg aglycones/day, together with about 25 g of soy protein has been suggested. For improvement in bone mineral density, 60-100 mg aglycones/day for a period of at least 6-12 months could be beneficial.


Biomechanical Evaluation of Rigid Interspinous Process Fixation Combined With Lumbar Interbody Fusion Using Hybrid Testing Protocol.
Rigid interspinous process fixation (RIPF) has been recently discussed as an alternative to pedicle screw fixation (PSF) for reducing trauma in lumbar interbody fusion (LIF) surgery. This study aimed to investigate biomechanics of the lumbar spine with RIPF, and also to compare biomechanical differences between two postoperative stages (before and after bony fusion). Based on an intact finite-element model of lumbosacral spine, the models of single-level LIF with RIPF or conventional PSF were developed and were computed for biomechanical responses to the moments of four physiological motions using hybrid testing protocol. It was found that compared with PSF, range of motion (ROM), intradiscal pressure (IDP), and facet joint forces (FJF) at adjacent segments of the surgical level for RIPF were decreased by up to 8.4%, 2.3%, and 16.8%, respectively, but ROM and endplate stress at the surgical segment were increased by up to 285.3% and 174.3%, respectively. The results of comparison between lumbar spine with RIPF before and after bony fusion showed that ROM and endplate stress at the surgical segment were decreased by up to 62.6% and 40.4%, respectively, when achieved to bony fusion. These findings suggest that lumbar spine with RIPF as compared to PSF has potential to decrease the risk of adjacent segment degeneration but might have lower stability of surgical segment and an increased risk of cage subsidence; When achieved bony fusion, it might be helpful for the lumbar spine with RIPF in increasing stability of surgical segment and reducing failure of bone contact with cage.


Two centuries of immunisation in the UK (part 1).
The impact of immunisation is best understood through a historical lens, since so many of the diseases which placed a burden on our population have been eliminated or controlled through immunisation. The United Kingdom (UK) National Health Service (NHS), which celebrated its 70th birthday in 2018, is responsible for delivering the highly successful universal national immunisation programme. However, the first vaccines used in the UK were not part of a centrally coordinated programme until the 1960s. Resources that summarise the first 200 years of immunisation in the UK are not readily accessible. Here we provide a two part chronological insight into the history of the UK immunisation programme from primary sources. In Part I, we highlight the importance of wartime conditions, unprecedented vaccine development, and the polio outbreaks in the in driving developments in immunisation and discuss subsequent changes in the use of the original vaccines of the immunisation programme, namely, diphtheria, tetanus, pertussis, and polio. In Part 2, we discuss the formation of the Joint Committee on Vaccination and Immunisation and its role, working with public health agencies and advising the UK Governments on vaccine policy, to bring a comprehensive programme to defend the health of the population against serious infectious diseases, highlighting the importance of programme organisation and leadership.


Synbiotic Combination of Djulis (Chenopodium formosanum) and Lactobacillus acidophilus Inhibits Colon Carcinogenesis in Rats.
Djulis is a functional grain containing prebiotic dietary fiber, which has an anti-cancer potential. This study examined the preventive effect of djulis alone or in combination with Lactobacillus acidophilus on colon carcinogenesis induced by 1,2-dimethylhydrazine (DMH) and dextran sulfate sodium (DSS). Rats were divided into five groups and fed B (AIN-93G, blank), C (AIN-93G, control), D (10% djulis), DLA (10% djulis plus 5 × 106 cfu L. acidophilus/g), and DHA (10% djulis plus 5 × 107 cfu L. acidophilus/g) diets, respectively. All rats except for those in group B received three doses of DMH (40 mg/kg) by intraperitoneal injection and 3% DSS in drinking water. After 10 weeks of feeding, the colon was analyzed for precancerous lesions and biomarkers. DMH and DSS treatment induced aberrant crypt foci (ACF), especially in the distal colon. D, DLA, and DHA significantly reduced the numbers of total ACF, sialomucin-producing ACF (SIM-ACF), and mucin-depleted foci (MDF) in the distal colon compared to C. Additionally, DLA and DHA further downregulated the expressions of proliferating cell nuclear antigen (PCNA) and cyclooxygenase-2 (COX-2) and regulated apoptosis-related proteins. These results suggest that synbiotic combination of djulis and L. acidophilus shows the best inhibitory effect on colon carcinogenesis via regulation of proliferative, inflammatory, and apoptotic pathways.


Automating gene library synthesis by structure-based combinatorial protein engineering: examples from plant sesquiterpene synthases.
Structure-based combinatorial protein engineering (SCOPE) is a homology-independent recombination method to create multiple crossover gene libraries by assembling defined combinations of structural elements ranging from single mutations to domains of protein structure. SCOPE was originally inspired by DNA shuffling, which mimics recombination during meiosis, where mutations from parental genes are "shuffled" to create novel combinations in the resulting progeny. DNA shuffling utilizes sequence identity between parental genes to mediate template-switching events (the annealing and extension of one parental gene fragment on another) in PCR reassembly reactions to generate crossovers and hence recombination between parental genes. In light of the conservation of protein structure and degeneracy of sequence, SCOPE was developed to enable the "shuffling" of distantly related genes with no requirement for sequence identity. The central principle involves the use of oligonucleotides to encode for crossover regions to choreograph template-switching events during PCR assembly of gene fragments to create chimeric genes. This approach was initially developed to create libraries of hybrid DNA polymerases from distantly related parents, and later developed to create a combinatorial mutant library of sesquiterpene synthases to explore the catalytic landscapes underlying the functional divergence of related enzymes. This chapter presents a simplified protocol of SCOPE that can be integrated with different mutagenesis techniques and is suitable for automation by liquid-handling robots. Two examples are presented to illustrate the application of SCOPE to create gene libraries using plant sesquiterpene synthases as the model system. In the first example, we outline how to create an active-site library as a series of complex mixtures of diverse mutants. In the second example, we outline how to create a focused library as an array of individual clones to distil minimal combinations of functionally important mutations. Through these examples, the principles of the technique are illustrated and the suitability of automating various aspects of the procedure for given applications are discussed.


Prevention of hepatitis C virus in hemodialysis patients: five years experience from a single center.
Hepatitis C virus (HCV) has been a significant problem in hemodialysis (HD) patients. In general, it carries significant morbidity including liver cirrhosis, liver cell failure and hepatoma. The study was conducted on 36 seronegative HD patients. All patients were managed with strict application of infection control guidelines as well as isolation of HCV-positive patients. None of the patients received any blood transfusions and were managed with iron and erythropoietin. After five years of follow-up, we found that the incidence of HCV seroconversion was zero. Our study further suggests that following infection control guidelines, isolation of seropositive patients and minimizing blood transfusions can help in prevention of HCV transmission among HD patients.


Deciphering the tête-à-tête between the microbiota and the immune system.
The past decade has witnessed an explosion in studies--both clinical and basic science--examining the relationship between the microbiota and human health, and it is now clear that the effects of commensal organisms are much broader than previously believed. Among the microbiota's major contributions to host physiology is regulation of the development and maintenance of the immune system. There are now a handful of examples of intestinal commensal bacteria with defined immunomodulatory properties, but our mechanistic understanding of how microbes influence the immune system is still in its infancy. Nevertheless, several themes have emerged that provide a framework for appreciating microbe-induced immunoregulation. In this Review, we discuss the current state of knowledge regarding the role of the intestinal microbiota in immunologic development, highlighting mechanistic principles that can guide future work.


MST and TRIC Technology to Reliably Study PROTAC Binary and Ternary Binding in Drug Development.
PROTACs have shown promise as a new class of therapy, with a unique mechanism of action orthogonal to traditional small molecules that are used to regulate protein activity. Their novel MOA utilizing the body's natural protein degradation machinery degrades a protein of interest rather than inhibiting its function. This strategy has several advantages over conventional small-molecule inhibitors, e.g., higher sensitivity, less off-target effects, and greater target space. However, unlocking the potential of PROTACs necessitates drug discovery techniques that can support the complexity of the novel MOA. In this chapter, we describe the application of MicroScale Thermophoresis (MST) and Temperature-Related Intensity Change (TRIC) to characterize both the binary and ternary binding of PROTACs with target proteins and ubiquitin ligases along with an efficient determination of the cooperativity of the ternary complex formation. The assay development and experimental procedure to characterize the well-described BET PROTAC MZ1 show how MST and TRIC can be applied as a fast and highly sensitive method for PROTAC discovery.


Reception and learning of electric fields in bees.
Honeybees, like other insects, accumulate electric charge in flight, and when their body parts are moved or rubbed together. We report that bees emit constant and modulated electric fields when flying, landing, walking and during the waggle dance. The electric fields emitted by dancing bees consist of low- and high-frequency components. Both components induce passive antennal movements in stationary bees according to Coulomb's law. Bees learn both the constant and the modulated electric field components in the context of appetitive proboscis extension response conditioning. Using this paradigm, we identify mechanoreceptors in both joints of the antennae as sensors. Other mechanoreceptors on the bee body are potentially involved but are less sensitive. Using laser vibrometry, we show that the electrically charged flagellum is moved by constant and modulated electric fields and more strongly so if sound and electric fields interact. Recordings from axons of the Johnston organ document its sensitivity to electric field stimuli. Our analyses identify electric fields emanating from the surface charge of bees as stimuli for mechanoreceptors, and as biologically relevant stimuli, which may play a role in social communication.


Use of admission Glasgow Coma Score, pupil size, and pupil reactivity to determine outcome for trauma patients.
Determination of nonsurvival in trauma patients is difficult because valid prognostic indicators are lacking. It was hypothesized that patients presenting with a Glasgow Coma Score (GCS) of 3 as well as fixed and dilated (FD) pupils do not have a reasonable chance of survival. From 1999 through 2001, adult trauma patients (age, >14 years) admitted with a GCS of 3 were reviewed. Patients receiving paralytic agents before initial assessment were excluded from analysis. Fixed and dilated pupils were defined as being 4 mm or more in diameter bilaterally and nonreactive to light. In this study, the FD patients were evaluated for survival, resuscitative measures, surgical procedures, length of hospital stay, and organ donation. The non-FD patients were evaluated for survival and length of hospital stay. Of the 137 patients evaluated with a GCS of 3, 104 had FD pupils and 33 did not. In the FD group, there were no survivors. On arrival, 28 (37.3%) of the patients were declared dead, and no further interventions were undertaken. Of the 76 patients (62.7%) who underwent further resuscitation, which included 53 surgical procedures, 30 died in the resuscitation bay, 39 within 24 hours, 4 within 48 hours, 2 within 72 hours, and 1 on day 6. There were 18 (23.7%) organ donors. Of the 33 patients without FD pupils, 11 (33%) survived to discharge (mean hospital stay, 21.4 days). Of the 22 nonsurvivors (67%), 10 died in the resuscitation bay, 8 within 24 hours, 1 within 48 hours, 1 on day 4, and 2 on day 6. Patients presenting with a GCS of 3 and FD pupils have no reasonable chance for survival. A significant percentage of these patients can be salvaged for organ donation. This information should be used in deciding to pursue aggressive resuscitation efforts and in discussing prognosis with family. Patients with a GCS of 3 who are not FD should be aggressively resuscitated because many of these patients survive to discharge.


NFC internal: an indoor navigation system.
Indoor navigation systems have recently become a popular research field due to the lack of GPS signals indoors. Several indoors navigation systems have already been proposed in order to eliminate deficiencies; however each of them has several technical and usability limitations. In this study, we propose NFC Internal, a Near Field Communication (NFC)-based indoor navigation system, which enables users to navigate through a building or a complex by enabling a simple location update, simply by touching NFC tags those are spread around and orient users to the destination. In this paper, we initially present the system requirements, give the design details and study the viability of NFC Internal with a prototype application and a case study. Moreover, we evaluate the performance of the system and compare it with existing indoor navigation systems. It is seen that NFC Internal has considerable advantages and significant contributions to existing indoor navigation systems in terms of security and privacy, cost, performance, robustness, complexity, user preference and commercial availability.


"Modeled microgravity" affects cell response to ionizing radiation and increases genomic damage.
The aim of this work was to assess whether "modeled microgravity" affects cell response to ionizing radiation, increasing the risk associated with radiation exposure. Lymphoblastoid TK6 cells were irradiated with various doses of gamma rays and incubated for 24 h in a modeled microgravity environment obtained by the Rotating Wall Vessel bioreactor. Cell survival, induction of apoptosis and cell cycle alteration were compared in cells irradiated and then incubated in 1g or modeled microgravity conditions. Modulation of genomic damage induced by ionizing radiation was evaluated on the basis of HPRT mutant frequency and the micronucleus assay. A significant reduction in apoptotic cells was observed in cells incubated in modeled microgravity after gamma irradiation compared with cells maintained in 1g. Moreover, in irradiated cells, fewer G2-phase cells were found in modeled microgravity than in 1g, whereas more G1-phase cells were observed in modeled microgravity than in 1g. Genomic damage induced by ionizing radiation, i.e. frequency of HPRT mutants and micronucleated cells, increased more in cultures incubated in modeled microgravity than in 1g. Our results indicate that modeled microgravity incubation after irradiation affects cell response to ionizing radiation, reducing the level of radiation-induced apoptosis. As a consequence, modeled microgravity increases the frequency of damaged cells that survive after irradiation.


Copeptin, a stable peptide derived from the vasopressin precursor, correlates with the individual stress level.
During stress, vasopressin is a potent synergistic factor of CRH as a hypothalamic stimulator of the HPA axis. The measurements of CRH and vasopressin levels are cumbersome because of their instability and short half-life. Copeptin is a more stable peptide stoichiometrically released from the same precursor molecule. The aim of our study was to compare copeptin and cortisol levels in different stress situations. Three groups of patients with increasing stress levels were investigated: a) healthy controls without apparent stress (n=20), b) hospitalized medical patients with moderate stress (n=25) and c) surgical patients 30 minutes after extubation, with maximal stress (n=29). In all patients we assessed cortisol and copeptin levels. Copeptin levels were measured with a new sandwich immunoassay. Cortisol levels in controls were (median, IQ range, 486 [397-588] nmol/L), not significantly different as compared to medical patients (438 [371-612] nmol/L, p=0.69). Cortisol levels in surgical patients after extubation were higher (744 [645-1062] nmol/L p<0.01 vs controls and medical patients). Copeptin levels in controls were 4.3 [3.2-5.5] pmol/L, which was lower as compared to medical patients (17.5 [6.4-24.1], p<0.001) and surgical patients after extubation (67.5 [37.8-110.0] pmol/L, p<0.001). The correlation between copeptin levels and cortisol was r=0.46, p<0.001. Copeptin is a novel marker of the individual stress level. It more subtly mirrors moderate stress as compared to cortisol values.


Intrinsic and extrinsic motivation is associated with computer-based auditory training uptake, engagement, and adherence for people with hearing loss.
Hearing aid intervention typically occurs after significant delay, or not at all, resulting in an unmet need for many people with hearing loss. Computer-based auditory training (CBAT) may provide generalized benefits to real-world listening, particularly in adverse listening conditions, and can be conveniently delivered in the home environment. Yet as with any intervention, adherence to CBAT is critical to its success. The main aim of this investigation was to explore motivations for uptake, engagement and adherence with home-delivered CBAT in a randomized controlled trial of adults with mild sensorineural hearing loss (SNHL), with a view to informing future CBAT development. A secondary aim examined perceived benefits of CBAT. Participants (n = 44, 50-74 years olds with mild SNHL who did not have hearing aids) completed a 4-week program of phoneme discrimination CBAT at home. Participants' experiences of CBAT were captured using a post-training questionnaire (n = 44) and two focus groups (n = 5 per group). A mixed-methods approach examined participants' experiences with the intervention, the usability and desirability of the CBAT software, and participants' motivations for CBAT uptake, engagement and adherence. Self-Determination Theory (SDT) was used as a theoretical framework for the interpretation of results. Participants found the CBAT intervention easy to use, interesting and enjoyable. Initial participation in the study was associated with extrinsic motivation (e.g., hearing difficulties). Engagement and adherence with CBAT was influenced by intrinsic (e.g., a desire to achieve higher scores), and extrinsic (e.g., to help others with hearing loss) motivations. Perceived post-training benefits included better concentration and attention leading to improved listening. CBAT also prompted further help-seeking behaviors for some individuals. We see this as an important first-step for informing future theory-driven development of effective CBAT interventions.


Deformations of promoter DNA bound to carcinogens help interpret effects on TATA-element structure and activity.
The TATA-box binding protein (TBP) is required by eukaryotic RNA polymerases for correct transcription initiation. TBP binds to the minor groove of an 8 base pair (bp) DNA-promoter element known as the TATA box and severely bends the TATA box. The promoter-DNA substrate can be damaged by components present in the cell or the environment to produce covalent carcinogen-DNA adducts. These may lead to transcription blockage or unfaithful transcription. Benzo[a]pyrene (BP) is a widespread environmental chemical carcinogen which can be metabolically converted to DNA-reactive enantiomeric (+) and (-)-anti-benzo[a]pyrene diol epoxides (BPDEs). Recent experimental studies of a pair of stereoisomeric adenine adducts, derived from (+) and (-)-anti-BPDEs, have revealed how these lesions influence the complexation of TBP with the TATA box. Depending on the adduct's location in the TATA box and its stereochemistry, the stability of monomeric TATA-TBP complexes was found to increase or decrease relative to the unmodified DNA. We report here analyses of molecular-dynamics simulations to interpret these findings. Structural analyses of 12 DNA-protein systems representing different combinations of adduct stereoisomer type and placement within the promoter reveal that the location of the adduct within the TATA octamer determines whether the stability of TATA-TBP complexes is increased or decreased. The effect on binding stability can be interpreted in terms of conformational freedom and major-groove space available to BP due to the hydrogen bonds and inserted phenylalanines of the TATA-TBP complex; that is, depending on the position of the adenine to which BP is covalently bound, BP can be accommodated in an intercalated or major-groove orientation with ease or with difficulty (due to interference with TATA-TBP interactions). The unravelled structures and interactions thus reveal the effect of different adduct locations on TATA-TBP complex formation and suggest how transcription initiation may be affected by the presence of a bulky BP.


Achieving cholesterol targets: how well are we doing?
Elevated low-density lipoprotein cholesterol (LDL-C) levels are a modifiable risk factor for the development of coronary heart disease (CHD), the leading cause of death in the United States. Treatments to lower these levels help decrease the risk of CHD events and reduce mortality rates in patients with existing CHD and those with no history of CHD. Rates of screening and treatment for high cholesterol levels have improved somewhat in recent years, but there is still room for substantial improvement, especially in patients at high risk of CHD, who benefit most from aggressive LDL-C-lowering therapies. The American Osteopathic Association Clinical Assessment Program, a Web-based program that measures physician performance by analyzing data abstracted from patient medical records and helps guide treatment decisions, is a tool to help physicians improve outcomes in patients with elevated LDL-C levels.


Therapeutic concepts and methods for improving dental implant outcomes. Summary and consensus statements. The 4th EAO Consensus Conference 2015.
Different therapeutic concepts and methods have been proposed for improving dental implant outcomes in three specific clinical situations: (i) the fresh extraction socket with alveolar ridge preservation protocols; (ii) the posterior maxilla with limited bone height with either the placement of regular-sized implants after sinus elevation and grafting or short dental implants and; (iii) the posterior mandible with limited bone height with either vertical bone augmentation and placement of implants or short dental implants. Three systematic reviews, based on randomized and controlled clinical trials have evaluated the efficacy of these different therapeutic modalities in terms of dental implant outcomes. Interventions aimed for alveolar ridge preservation have shown efficacy in terms of allowing the placement of dental implants and for reducing the need of further augmentation procedures at implant placement. Both therapeutic options, the placement of implants after sinus elevation and grafting or short dental implants, were valid alternatives in the treatment of the posterior maxilla with deficient bone availability, although short implants resulted in fewer complications. Similarly, the placement of implants in vertically augmented bone rendered comparable outcomes with those of short implants in the treatment of the posterior mandible, but short implants resulted in fewer complications.


[A case of limbic encephalitis associated with breast cancer developed in an HTLV-1 carrier].
We here present a case of 44-year-old woman, a carrier of human T-lymphotrophic virus type-1 (HTLV-1), who suffered from limbic encephalitis and breast cancer. In December 1997, the patient's behavior became abnormal. Three weeks later, she became markedly forgetful. At that time neurological examinations revealed that she had anterograde and retrograde amnesia, disorientation, and confabulation, although her consciousness was clear. Anti-Hu and anti-Yo antibodies and antinuclear antibodies in the serum were negative. Flow cytometric study of the peripheral blood lymphocytes showed an increased percentage of CD3+CD25+ cells, although the percentages of CD4+CD45RA+ and CD4+CD45RO+ cells were normal. Lymphocytic responses to phytohemagglutinin or concanavalin A were normal. Anti-HTLV-1 antibody was positive both in the serum and in the cerebrospinal fluid (CSF). The level of immunoglobulin G was high and two oligoclonal immunoglobulin G bands were positive in the CSF. Cytological study of the CSF showed no atypical cells. Findings for herpes simplex virus type I and II DNAs were negative with polymerase chain reaction in the CSF. There was no elevation of antibody titers against viruses including herpes simplex virus, cytomegalovirus, and measles virus, either in the serum or the CSF. Magnetic resonance imaging showed signal abnormalities in the medial portions of both temporal lobes, in particular, in the bilateral hippocampus. Six weeks after the onset, a cancerous tumor in her right breast was detected and removed by open surgery. The pathological diagnosis was invasive ductal carcinoma with neuroendocrine features. After mastectomy, anterograde and retrograde amnesia and disorientation mildly improved. The follow-up magnetic resonance imaging showed that signal abnormalities in the medial portions of both temporal lobes decreased and that the bilateral hippocampus became atrophic. We diagnosed the present case as paraneoplastic limbic encephalitis. There has been only one case report of limbic encephalitis associated with breast cancer.


Estimation of the postmortem duration of mouse tissue by electron spin resonance spectroscopy.
Electron spin resonance (ESR) method is a simple method for detecting various free radicals simultaneously and directly. However, ESR spin trap method is unsuited to analyze weak ESR signals in organs because of water-induced dielectric loss (WIDL). To minimize WIDL occurring in biotissues and to improve detection sensitivity to free radicals in tissues, ESR cuvette was modified and used with 5,5-dimethtyl-1-pyrroline N-oxide (DMPO). The tissue samples were mouse brain, hart, lung, liver, kidney, pancreas, muscle, skin, and whole blood, where various ESR spin adduct signals including DMPO-ascorbyl radical (AsA(∗)), DMPO-superoxide anion radical (OOH), and DMPO-hydrogen radical (H) signal were detected. Postmortem changes in DMPO-AsA(∗) and DMPO-OOH were observed in various tissues of mouse. The signal peak of spin adduct was monitored until the 205th day postmortem. DMPO-AsA(∗) in liver (y = 113.8-40.7 log (day), R1 = -0.779, R2 = 0.6, P < .001) was found to linearly decrease with the logarithm of postmortem duration days. Therefore, DMPO-AsA(∗) signal may be suitable for detecting an oxidation stress tracer from tissue in comparison with other spin adduct signal on ESR spin trap method.


The biochemistry of apoptosis.
Apoptosis--the regulated destruction of a cell--is a complicated process. The decision to die cannot be taken lightly, and the activity of many genes influence a cell's likelihood of activating its self-destruction programme. Once the decision is taken, proper execution of the apoptotic programme requires the coordinated activation and execution of multiple subprogrammes. Here I review the basic components of the death machinery, describe how they interact to regulate apoptosis in a coordinated manner, and discuss the main pathways that are used to activate cell death.


Structural Basis for the Binding of Allosteric Activators Leucine and ADP to Mammalian Glutamate Dehydrogenase.
Glutamate dehydrogenase (GDH) plays a key role in the metabolism of glutamate, an important compound at a cross-road of carbon and nitrogen metabolism and a relevant neurotransmitter. Despite being one of the first discovered allosteric enzymes, GDH still poses challenges for structural characterization of its allosteric sites. Only the structures with ADP, and at low (3.5 Å) resolution, are available for mammalian GDH complexes with allosteric activators. Here, we aim at deciphering a structural basis for the GDH allosteric activation using bovine GDH as a model. For the first time, we report a mammalian GDH structure in a ternary complex with the activators leucine and ADP, co-crystallized with potassium ion, resolved to 2.45 Å. An improved 2.4-angstrom resolution of the GDH complex with ADP is also presented. The ternary complex with leucine and ADP differs from the binary complex with ADP by the conformation of GDH C-terminus, involved in the leucine binding and subunit interactions. The potassium site, identified in this work, may mediate interactions between the leucine and ADP binding sites. Our data provide novel insights into the mechanisms of GDH activation by leucine and ADP, linked to the enzyme regulation by (de)acetylation.


Modeling of coupled-resonator optical waveguide (CROW) based refractive index sensors using pixelized spatial detection at a single wavelength.
We model and analyze coupled-resonator optical waveguide (CROW) based refractive index (RI) sensors using pixelized spatial detection. Our modeled cascaded Fabry-Perot (FP) CROWs reveal that the intra-band states mode-field distributions vary upon effective RI change at a single wavelength. The spatial Fourier transform of the CROW mode-field distributions, with each cavity field intensity integrated as a pixel, shows spatial frequency peak shift, which constitutes the basis of such a spatial domain sensor. The spatial domain sensing performance depends on the cavity number, the cavity length and the inter-cavity coupling. Our modeled 21-element CROW sensor attains a detection limit of 10(-4) refractive index unit (RIU) with a sensing dynamic range of 10(-3) RIU. Detailed analysis of the spatial frequency harmonic peak amplitude variation further suggests an improved detection limit. Finite-difference time-domain (FDTD) simulations of an 11-element microring CROW device shows sensitivity consistent with the FP modeling.


Evolutionarily conserved protein sequences of influenza a viruses, avian and human, as vaccine targets.
Influenza A viruses generate an extreme genetic diversity through point mutation and gene segment exchange, resulting in many new strains that emerge from the animal reservoirs, among which was the recent highly pathogenic H5N1 virus. This genetic diversity also endows these viruses with a dynamic adaptability to their habitats, one result being the rapid selection of genomic variants that resist the immune responses of infected hosts. With the possibility of an influenza A pandemic, a critical need is a vaccine that will recognize and protect against any influenza A pathogen. One feasible approach is a vaccine containing conserved immunogenic protein sequences that represent the genotypic diversity of all current and future avian and human influenza viruses as an alternative to current vaccines that address only the known circulating virus strains. Methodologies for large-scale analysis of the evolutionary variability of the influenza A virus proteins recorded in public databases were developed and used to elucidate the amino acid sequence diversity and conservation of 36,343 sequences of the 11 viral proteins of the recorded virus isolates of the past 30 years. Technologies were also applied to identify the conserved amino acid sequences from isolates of the past decade, and to evaluate the predicted human lymphocyte antigen (HLA) supertype-restricted class I and II T-cell epitopes of the conserved sequences. Fifty-five (55) sequences of 9 or more amino acids of the polymerases (PB2, PB1, and PA), nucleoprotein (NP), and matrix 1 (M1) proteins were completely conserved in at least 80%, many in 95 to 100%, of the avian and human influenza A virus isolates despite the marked evolutionary variability of the viruses. Almost all (50) of these conserved sequences contained putative supertype HLA class I or class II epitopes as predicted by 4 peptide-HLA binding algorithms. Additionally, data of the Immune Epitope Database (IEDB) include 29 experimentally identified HLA class I and II T-cell epitopes present in 14 of the conserved sequences. This study of all reported influenza A virus protein sequences, avian and human, has identified 55 highly conserved sequences, most of which are predicted to have immune relevance as T-cell epitopes. This is a necessary first step in the design and analysis of a polyepitope, pan-influenza A vaccine. In addition to the application described herein, these technologies can be applied to other pathogens and to other therapeutic modalities designed to attack DNA, RNA, or protein sequences critical to pathogen function.


Pepfar 3.0's HIV testing policy in Côte d'Ivoire (2014 to 2018): fragmentation, acceleration and disconnection.
HIV Testing and Counselling (HTC) remains a key challenge in achieving control of the HIV epidemic by 2030. In the early 2010s, the President's Emergency Plan for AIDS Relief (Pepfar) adopted targeted HTC strategies for populations and geographical areas most affected by HIV. We examine how Pepfar defined targeted HTC in Côte d'Ivoire, a country with a mixed HIV epidemic, after a decade of expanding HTC services. We explored the evolution of HTC strategies through the Country Operational Plans (COP) of Pepfar during its phase 3.0, from COP 14 to COP 17 (October 2014 to September 2018) in Côte d'Ivoire. We conducted an analysis of the grey literature over the period 2014 to 2018 (Budget & Target Report, Strategic Direction Summary, Sustainability Index and Dashboard Summary, https://data.pepfar.gov). We also conducted a qualitative study in Côte d'Ivoire (2015 to 2018) using in-depth interviews with stakeholders in the AIDS public response: CDC/Pepfar (3), Ministry of Health (3), intermediary NGOs (7); and public meeting observations (14). Since the COP 14, Pepfar's HIV testing strategies have been characterized by significant variations in terms of numerical, geographical and population targets. While the aim of COP 14 and COP 15 seemed to be the improvement of testing efficacy in general and testing yield in particular, COP 16 and COP 17 prioritized accelerating progress towards the "first 90" (i.e. reducing the proportion of people living with HIV who are unaware of their HIV). A shift was observed in the definition of testing targets, with less focus on the inclusion of programmatic data and feedback from field actors, and greater emphasis on the use of models to estimate and disaggregate the targets by geographical units and sub-populations (even if the availability of data by this disaggregation was limited or uncertain); increasingly leading to gaps between targets and results. These trials and tribulations question the real and long-term effectiveness of annually-revised, fragmented strategies, which widen an increasing disparity between the realities of the actors on the ground and the objectives set in Washington.


First detection of Borrelia burgdorferi sensu lato DNA in king penguins (Aptenodytes patagonicus halli).
The hard tick Ixodes uriae parasitises a wide range of seabird species in the circumpolar areas of both Northern and Southern hemispheres and has been shown to be infected with Borrelia burgdorferi sensu lato, the bacterial agents of Lyme borreliosis. Although it is assumed that seabirds represent viable reservoir hosts, direct demonstrations of infection are limited to a single study from the Northern hemisphere. Here, the blood of 50 tick-infested adult king penguins (Aptenodytes patagonicus halli) breeding in the Crozet Archipelago (Southern Indian Ocean) was examined for B. burgdorferi sl exposure by serology and for spirochetemia by in vitro DNA amplification. Four birds were found positive by serology, whereas B. burgdorferi sl DNA was detected in two other birds. Our data therefore provide the first direct proof of Borrelia burgdorferi sl spirochetes in seabirds of the Southern hemisphere and indicate a possible reservoir role for king penguins in the natural maintenance of this bacterium. Although the bacterial genetic diversity present in these hosts and the infectious period for tick vectors remain to be elucidated, our results add to a growing body of knowledge on the contribution of seabirds to the complex epizootiology of Lyme disease and the global dissemination of B. burgdorferi sl spirochetes.


Effectiveness and Safety of Apixaban Versus Warfarin Among Older Patients with Venous Thromboembolism with Different Demographics and Socioeconomic Status.
Impact of demographics and socioeconomic status (SES) on anticoagulant treatment outcomes among patients with venous thromboembolism (VTE) is not well understood. This study evaluated risks of recurrent VTE, major bleeding (MB), and clinically relevant non-major bleeding (CRNMB) among older patients with VTE initiating apixaban or warfarin stratified by demographics and SES. Adult patients (≥ 65 years) who initiated apixaban or warfarin after a VTE event were selected from the US CMS Medicare database (September 2014-December 2017). Stabilized inverse probability treatment weighting (IPTW) was used to balance patient characteristics between treatment cohorts. Patients were stratified by age, gender, race, and SES. For each subgroup, Cox proportional hazard models were used to evaluate if there was a significant interaction (p < 0.10) between treatment and subgroup for recurrent VTE, MB, and CRNMB. In total, 22,135 apixaban and 45,840 warfarin patients with VTE were included. Post-IPTW, patient characteristics were balanced between treatment cohorts. In older patients, apixaban treatment was associated with significantly lower risks of recurrent VTE (hazard ratio [HR] 0.64; 95% confidence interval [CI] 0.52-0.79), MB (HR 0.65; 95% CI 0.57-0.75), and CRNMB (HR 0.79; 95% CI 0.75-0.85) versus warfarin. When stratified by demographics and SES, higher incidence rates of recurrent VTE, MB, and CRNMB were observed for black vs white patients and patients with lower vs higher SES. Comparison of apixaban with warfarin by different demographic and SES subgroups showed generally consistent results as the overall analysis. For most subgroups, no significant interaction was observed between treatment and subgroup strata for recurrent VTE, MB, and CRNMB. Among older patients with VTE initiating apixaban or warfarin, higher rates of recurrent VTE and bleeding were observed in black patients and patients with lower SES. Apixaban had a lower risk of recurrent VTE, MB, and CRNMB compared to warfarin. Analyses of demographic and SES subgroups showed consistent findings.


Maximal exercise cardiorespiratory responses of men and women during acute exposure to hypoxia.
Three male and four female subjects were acutely exposed to normoxic and hypoxic gas mixtures (FIO2 = 17.39%, 14.40%, 11.81%) in a single-blind randomized fashion during four treadmill runs to volitional exhaustion. Maximal scores for oxygen consumption (VO2), carbon dioxide production and heart rate decreased linearly (p less than 0.01) with increasing hypoxia. Conversely, maximal scores for ventilation, ventilatory equivalent (VE/VO2) and R increased linearly (p less than 0.01) with decreasing FIO2. During hypoxia, no significant differences in work time or respiratory compensation threshold were evident. However, female subjects had significantly higher (p less than 0.05) VE/VO2 scores and showed a relative decrease in VO2max that was significantly less (p less than 0.01) than male subjects. It was concluded that young highly active females, when compared to males of similar age and relative condition, have a stronger adaptive response to acute hypoxia during a maximal treadmill run.


Evaluation of a revised fourth grade social problem solving curriculum: Empathy as a moderator of adjustive gain.
Two classrooms of fourth grade suburban children (n=37) were trained using a 20-lesson revised Social Problem Solving (SPS) curriculum and compared to two no-training comparison classrooms (n=45). Results indicated a significant increase in the number of alternative solutions to interpersonal difficulties generated by E versus C children on a group social problem solving measure. Adjustive gains for E versus C children were reported by teachers and children on indices of school adjustment. These findings supported the efficacy of the shortened curriculum in increasing problem solving skills and facilitating positive adjustive change.The link between gain in SPS skills and adjustive change was examined in two ways. Delta scores compared changes in SPS skills and changes in teacher/child adjustive ratings yield no significant relationships. Hierarchical regression analyses, incorporating a moderator variable (empathy), indicated a significant relationship among pre level of empathy, gain in SPS skills and teacher rated adjustive change. These findings supported the hypothesis that key moderating variables may play a role in the training and employment of SPS skills and that strategies designed to examine these variables need to be employed.


Anatomy of the human mammary gland: Current status of knowledge.
Mammary glands are unique to mammals, with the specific function of synthesizing, secreting, and delivering milk to the newborn. Given this function, it is only during a pregnancy/lactation cycle that the gland reaches a mature developmental state via hormonal influences at the cellular level that effect drastic modifications in the micro- and macro-anatomy of the gland, resulting in remodeling of the gland into a milk-secretory organ. Pubertal and post-pubertal development of the breast in females aids in preparing it to assume a functional state during pregnancy and lactation. Remarkably, this organ has the capacity to regress to a resting state upon cessation of lactation, and then undergo the same cycle of expansion and regression again in subsequent pregnancies during reproductive life. This plasticity suggests tight hormonal regulation, which is paramount for the normal function of the gland. This review presents the current status of knowledge of the normal macro- and micro-anatomy of the human mammary gland and the distinct changes it undergoes during the key developmental stages that characterize it, from embryonic life through to post-menopausal age. In addition, it discusses recent advances in our understanding of the normal function of the breast during lactation, with special reference to breastmilk, its composition, and how it can be utilized as a tool to advance knowledge on normal and aberrant breast development and function. Finally, anatomical and molecular traits associated with aberrant expansion of the breast are discussed to set the basis for future comparisons that may illuminate the origin of breast cancer.


Fractional exhaled nitric oxide for the management of asthma in adults: a systematic review.
The aim of this review was to evaluate the clinical effectiveness of fractional exhaled nitric oxide (FeNO) measured in a clinical setting for the management of asthma in adults.13 electronic databases were searched and studies were selected against predefined inclusion criteria. Quality assessment was conducted using QUADAS-2. Class effect meta-analyses were performed.Six studies were included. Despite high levels of heterogeneity in multiple study characteristics, exploratory class effect meta-analyses were conducted. Four studies reported a wider definition of exacerbation rates (major or severe exacerbation) with a pooled rate ratio of 0.80 (95% CI 0.63-1.02). Two studies reported rates of severe exacerbations (requiring oral corticosteroid use) with a pooled rate ratio of 0.89 (95% CI 0.43-1.72). Inhaled corticosteroid use was reported by four studies, with a pooled standardised mean difference of -0.24 (95% CI -0.56-0.07). No statistically significant differences for health-related quality of life or asthma control were found.FeNO guided management showed no statistically significant benefit in terms of severe exacerbations or inhaled corticosteroid use, but showed a statistically significant reduction in exacerbations of any severity. However, further research is warranted to clearly define which management protocols (including cut-off points) offer best efficacy and which patient groups would benefit the most.


Evaluation of left ventricular mass and function, lipid profile, and insulin resistance in Egyptian children with growth hormone deficiency: A single-center prospective case-control study.
Growth hormone deficiency (GHD) in adults is associated with a cluster of cardiovascular risk factors that may contribute to an increased mortality for cardiovascular disease. In children, relatively few studies have investigated the effect of GHD and replacement therapy on cardiac performance and metabolic abnormalities that may place them at a higher risk of cardiovascular disease (CVD) at an early age. This study was aimed to assess the left ventricular function, lipid profile, and degree of insulin resistance in Egyptian children with GHD before and after 1 year of GH replacement therapy. Prospective case-control study, single-center study. Thirty children with short stature due to GHD were studied in comparison to 20 healthy age- and sex-matched children. All subjects were subjected to history, clinical examination, auxological assessment, and echocardiography to assess the left ventricular function. Blood samples were collected for measuring IGF-1, lipid profile (Total, LDL, HDL cholesterol, triglyceride, and atherogenic index (AI), fasting blood sugar, and fasting insulin levels. In addition, basal and stimulated GH levels were measured in children with suspected GHD. Student's t-test was used for parametric data, and the Mann-Whitney U-test was used for non-parametric data. Total, LDL cholesterol, triglyceride, AI, and insulin were significantly higher in children with GHD than in healthy controls at baseline. After 12 months of GH replacement therapy, total, LDL cholesterol, triglyceride, AI and insulin were significantly decreased, while homeostatic model assessment for insulin resistance index (HOMA-IR) was significantly increased compared to both pre-treatment and control values. At baseline, the left ventricular mass (LVM) and left ventricular mass index (LVMi) were significantly lower in GHD children than in controls. After 12 months of GH replacement therapy, LVM and LVMi in GHD patients were significantly increased compared to pre-treatment values. GHD in children is associated with a significantly reduced cardiac mass and impairment of lipid profile. GH replacement therapy exerts beneficial effects both on cardiac mass and lipid metabolism by normalizing cardiac size and improving the lipid profile. On the contrary, an increase in insulin resistance is observed after 12 months GH treatment. The study suggests that children with GH deficiency should have echocardiography and lipid profile monitoring before and during treatment with GH.


Radiation exposure during endovascular procedures.
The duration of fluoroscopy exposure is routinely recorded as part of endovascular procedures. However, to better relate the duration of exposure to actual doses of surface and intracranial radiation, we compared surface doses during endovascular procedures with intracranial doses in a cadaver model exposed to lateral fluoroscopy. Optically stimulated luminescence dosimeter chips (Landauer, Glenwood, IL) were used to measure the cranial surface dose of three consecutive patients undergoing endovascular procedures. Bitemporal craniotomies were performed on a cadaver. Dosimeter chips were placed on both the ipsilateral and contralateral skin and meningeal surfaces, and the cadaver was exposed to lateral fluoroscopy. Finally, to assess mean fluoroscopy times in patients undergoing embolization procedures, the operative notes of 100 consecutive patients were reviewed. Three patients undergoing endovascular treatment received peak doses of 0.24, 0.31, and 1.38 Gy, respectively. In the cadaver, the peak surface dose recorded after 120 minutes of exposure was 1.71 Gy. The cranium and scalp absorbed or reflected 29% of the surface dose. Time in minutes of fluoroscopy was found to correlate with surface dose (R2 = 0.925). Our data show that radiation exposure during endovascular treatment can reach clinically significant levels. The surface doses recorded during this study were comparable to the mean dose of 1.5 Gy estimated by others to increase the relative risk of inducing meningiomas, gliomas, and nerve sheath tumors. Pending long-term follow-up of patients exposed to endovascular procedures, consent for possible long-term sequelae of radiation may be warranted.


Rapid detection of fungal keratitis with DNA-stabilizing FTA filter paper.
Purpose. Polymerase chain reaction (PCR) is increasingly important for the rapid detection of fungal keratitis. However, techniques of specimen collection and DNA extraction before PCR may interfere with test sensitivity. The purpose of this study was to investigate the use of DNA-stabilizing FTA filter paper (Indicating FTA filter paper; Whatman International, Ltd., Maidstone, UK) for specimen collection without DNA extraction in a single-step, nonnested PCR for fungal keratitis. Methods. Specimens were collected from ocular surfaces with FTA filter discs, which automatically lyse collected cells and stabilize nucleic acids. Filter discs were directly used in single-step PCR reactions to detect fungal DNA. Test sensitivity was evaluated with serial dilutions of Candida albicans, Fusarium oxysporum, and Aspergillus fumigatus cultures. Test specificity was analyzed by comparing 196 and 155 healthy individuals from Switzerland and Egypt, respectively, with 15 patients with a diagnosis of microbial keratitis. Results. PCR with filter discs detected 3 C. albicans, 25 F. oxysporum, and 125 A. fumigatus organisms. In healthy volunteers, fungal PCR was positive in 1.0% and 8.4% of eyes from Switzerland and Egypt, respectively. Fungal PCR remained negative in 10 cases of culture-proven bacterial keratitis, became positive in 4 cases of fungal keratitis, but missed 1 case of culture-proven A. fumigatus keratitis. Conclusions. FTA filter paper for specimen collection together with direct PCR is a promising method of detecting fungal keratitis. The analytical sensitivity is high without the need for a semi-nested or nested second PCR, the clinical specificity is 91.7% to 99.0%, and the method is rapid and inexpensive.


Human interleukin 1. Purification to homogeneity.
We have purified human interleukin 1 (IL-1) to homogeneity by a simplified procedure that results in excellent yields of pure material that retains a high level of biological activity. IL-1, secreted by human peripheral blood macrophages that have been stimulated with Staphylococcus aureus, was purified by ion exchange chromatography and affinity chromatography on Procion Red agarose. The pure protein has a specific activity of 3.2 X 10(8) U/mg in the thymocyte mitogenesis assay, and is pyrogenic. No molecular weight heterogeneity was observed, in contrast to findings for mouse IL-1 and earlier reports of human IL-1. Purified IL-1, as analyzed by two-dimensional electrophoresis/electrofocusing gels, exhibited a series of charged species with isoelectric points ranging from 6.0 to 4.9, all with a molecular weight of approximately 17,500. Amino acid analysis indicated an abundance of acidic residues, in agreement with the low isoelectric points. There is little or no cysteine in the molecule. No evidence was found for the presence of carbohydrate moieties. The overall yield for this procedure was approximately 31% of the activity contained in the initial culture supernatant.